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McCabe Dunn J.M.,Merck And Co. | Reibarkh M.,Merck And Co. | Sherer E.C.,Merck And Co. | Orr R.K.,Merck And Co. | And 3 more authors.
Chemical Science | Year: 2017

The direct and chemoselective 3′-phosphoramidation, phosphorylation and acylation of nucleosides are described. Upon the discovery of a novel 3′-phosphorylamidation of therapeutic nucleoside analogues with DBU, we explored the mechanism of this rare selectivity through a combination of NMR spectroscopy and computational studies. The NMR and computational findings allowed us to develop a predictive computational model that accurately assesses the potential for 3′-functionalization for a broad range of nucleosides and nucleoside mimetics. The synthetic utility of this model was exemplified by demonstration on a broad scope of nucleosides and electrophiles yielding targets that were previously only accessible via a protection/deprotection sequence or an enzymatic approach. © The Royal Society of Chemistry.

Zhang C.,University of Utah | Tutkowski B.,University of Notre Dame | DeLuca R.J.,University of Utah | Joyce L.A.,Merck And Co. | And 2 more authors.
Chemical Science | Year: 2017

An enantioselective, redox-relay Heck alkenylation of trisubstituted allylic alkenol substrates has been developed. This process enables the construction of vicinal stereocenters in high diastereo- and enantioselectivity and allows the formation of enolizable α-carbonyl methyl-substituted stereocenters with no observed epimerization under the reported reaction conditions. © The Royal Society of Chemistry.

Yang F.,Merck And Co. | Brown C.D.,Merck And Co. | Rosen L.A.,Merck And Co.
Annual Technical Conference - ANTEC, Conference Proceedings | Year: 2016

Despite significant advances have been achieved in applying amorphous solid dispersion to enhance bioavailability of poorly soluble active pharmaceutical ingredients (APIs), there remain challenges in characterizing the microstructures of solid dispersions and correlating their performance with microstructures. This study focused on utilizing rheology as a tool to investigate and evaluate several model polymer/API solid dispersions prepared by various techniques with different mixing capabilities. Rheological responses of different model solid dispersions displayed a strong correlation between microstructures and viscoelastic properties. For the currently studied system, storage modulus and viscosities versus frequency of different solid dispersions indicated that the incorporation of API imparted a plasticizing effect to the polymer matrix. In comparison, crystalline/aggregated forms of the API exhibited a more elastic response than its amorphous/dispersed counterparts. In addition, a temperature ramp interrogation of a physical mixture of polymer and API captured a critical temperature, at which a transition in slope observed in the damping factor was attributed to the dissolution of crystalline API into the polymer.

Yang F.,Merck And Co. | Brown C.D.,Merck And Co.
Annual Technical Conference - ANTEC, Conference Proceedings | Year: 2016

The production of amorphous solid dispersions via hot melt extrusion (HME) relies on elevated temperature, applied mechanical force and prolonged residence time, which can result in potential degradation and decomposition of thermally-sensitive components. In this study, the rheological properties of a physical mixture of polymer/active pharmaceutical ingredient (API) were utilized to guide HME processing temperature. A critical temperature, which is substantially lower (~13oC) than the melting point of crystalline API, was captured during a temperature ramp examination and regarded as the critical point at which the API molecularly dissolves into the polymer. After identification, solid dispersions were prepared by HME processing below, on, or above the recognized critical temperature and characterized by scanning electron microscopy, hot stage microscopy, Xray diffraction, differential scanning calorimetry and rheology. Physicochemical properties of resultant solid dispersions indicated that the obtained critical temperature is sufficient for the polymer/API system to reach a molecular-level mixing, manifested by the transparent and smooth appearance of extrudates, absence of API crystalline diffraction and melting peaks, and dramatically decreased complex viscosity. Once the critical temperature is achieved, further raising temperature only results in limited improvement of the dispersion, reflected by slightly reduced storage modulus and complex viscosity.

DiNubile M.J.,Merck And Co.
Open Forum Infectious Diseases | Year: 2016

Novel treatment options are urgently needed for patients with serious multidrug-resistant infections seen increasingly in routine everyday clinical practice, both in the hospital and nursing home as well as in the clinic and office setting. Unfortunately, the problem is no longer confined to chronically ill, repeatedly hospitalized patients. This essay explores the role of noninferiorly studies in addressing the pressing need for new antimicrobial agents to combat the emerging "superbugs", calling attention to the nuances of interpreting their sometimes less-than-straightforward results. The overriding aim is not to find better antibiotics for routinely treatable infections but to identify safe and efficacious treatment options where none presently exist. © The Author 2016.

Thaisrivongs D.A.,Merck And Co. | Naber J.R.,Merck And Co. | McMullen J.P.,Merck And Co.
Organic Process Research and Development | Year: 2016

We report that an organolithium addition to a chiral ketimine, which because of competitive proton transfer proceeds with only modest yield in batch, can be significantly improved in flow. The transformation was discovered to be highly mixing-sensitive but remarkably temperature-independent in flow, enabling the continuous process to be performed under noncryogenic conditions. Experiments at the kilogram scale provided robust operating conditions that are amenable to implementing this chemistry for the manufacture of verubecestat (MK-8931), a drug candidate for the treatment of Alzheimer's disease. © 2016 American Chemical Society.

Du M.,Evidera | Chase M.,Merck And Co. | Oguz M.,Evidera | Davies G.,Merck And Co.
Current Medical Research and Opinion | Year: 2017

Objective: To evaluate long-term health benefits and risks of adding vorapaxar (VOR) to the standard care antiplatelet therapy (SC) of aspirin and/or clopidogrel, among a population with a recent myocardial infarction (MI) and/or peripheral artery disease (PAD). Research design and methods: In a state-transition model, patients transition between health states (event-free, recurrent MI, stroke, death), while at risk of experiencing non-transition-related revascularization and non-fatal bleeding events. Risk equations developed from the TRA 2°P-TIMI 50 trial’s patient-level data were used to predict cardiovascular (CV) outcomes over longer time horizons. Additional sources, including trials and US-based observational studies, informed the inputs for short-term CV risk, non-CV death, and health-related quality of life. Survival and quality-adjusted life-years (QALYs) were estimated over a lifetime horizon, discounted at 3% per year. Results: Within a cohort of 7361 patients with recent MI and/or PAD, VOR + SC relative to SC alone yielded 176 fewer CV events (MIs, strokes, or CV deaths), but 27 more major bleeding events. VOR + SC was associated with increased life expectancy and health benefits (19.93 undiscounted life-years [LYs], 9.57 discounted QALYs vs. 19.61 undiscounted LYs, 9.41 discounted QALYs). The results were most sensitive to scenarios varying time of vorapaxar initiation, and the assumptions in the 90 day period post-MI. Additional analyses showed that add-on vorapaxar provides consistent incremental benefits in high-risk subgroups. Conclusion: This study contributes to the growing literature on secondary prevention add-on therapy, as results from these modeling analyses suggest that adding vorapaxar to SC for patients at high atherothrombotic risk can provide long-term health benefits. © 2017 Informa UK Limited, trading as Taylor & Francis Group

Filzen T.M.,Merck And Co. | Kutchukian P.S.,Merck And Co. | Hermes J.D.,Merck And Co. | Li J.,Merck And Co. | Tudor M.,Merck And Co.
PLoS Computational Biology | Year: 2017

High throughput mRNA expression profiling can be used to characterize the response of cell culture models to perturbations such as pharmacologic modulators and genetic perturbations. As profiling campaigns expand in scope, it is important to homogenize, summarize, and analyze the resulting data in a manner that captures significant biological signals in spite of various noise sources such as batch effects and stochastic variation. We used the L1000 platform for large-scale profiling of 978 representative genes across thousands of compound treatments. Here, a method is described that uses deep learning techniques to convert the expression changes of the landmark genes into a perturbation barcode that reveals important features of the underlying data, performing better than the raw data in revealing important biological insights. The barcode captures compound structure and target information, and predicts a compound’s high throughput screening promiscuity, to a higher degree than the original data measurements, indicating that the approach uncovers underlying factors of the expression data that are otherwise entangled or masked by noise. Furthermore, we demonstrate that visualizations derived from the perturbation barcode can be used to more sensitively assign functions to unknown compounds through a guilt-by-association approach, which we use to predict and experimentally validate the activity of compounds on the MAPK pathway. The demonstrated application of deep metric learning to large-scale chemical genetics projects highlights the utility of this and related approaches to the extraction of insights and testable hypotheses from big, sometimes noisy data. © 2017 Filzen et al.

Ahmed S.E.,Brock University | Hussein A.,University of Windsor | Ghori R.,Merck And Co.
Advances in Intelligent Systems and Computing | Year: 2017

The process capability indices (PCI) have been popular in the manufacturing environment to quantify the capability of an industrial process. In this paper, a robust estimator of the Cpindex, based on the Gini mean difference statistic, is proposed. The performance of the proposed estimator and its associated confidence intervals are compared to those associated with the classical estimator based on the sample standard deviation. The use of the new method is illustrated by application to data set about membrane thickness of STN color pixels. © Springer Science+Business Media Singapore 2017.

Wassermann A.M.,Merck And Co. | Tudor M.,Merck And Co. | Glick M.,Merck And Co.
Drug Discovery Today: Technologies | Year: 2017

The term dark chemical matter (DCM) was recently introduced for those molecules in a screening collection that have never shown any substantial biological activity despite having been tested in hundreds of high-throughput assays. It was suggested that, if hits emerge from this compound pool in future screening campaigns, they should be prioritized due to their exquisite selectivity profile. In this article we define DCM at our company and describe on-going efforts to shed light on the bioactivity of these apparently silent compounds, with an emphasis on multi-parametric profiling methods. It is also demonstrated that compounds that are dark within one institution might be found active in another, but typically show the foretold selectivity. © 2017 Elsevier Ltd.

The process of developing international (ICH) guidelines is described, and the main guidelines reviewed are the ICH S2(R1) guideline that includes the genotoxicity test battery for human pharmaceuticals, and the ICH M7 guideline for assessing and limiting potentially mutagenic impurities and degradation products in drugs. Key aspects of the guidelines are reviewed in the context of drug development, for example the incorporation of genotoxicity assessment into non-clinical toxicity studies, and ways to develop and assess weight of evidence. In both guidelines, the existence of "thresholds" or non-linear dose responses for genotoxicity plays a part in the strategies. Differences in ICH S2(R1) protocol recommendations from OECD guidelines are highlighted and rationales explained. The use of genotoxicity data during clinical development and in assessment of carcinogenic potential is also described. There are no international guidelines on assessment of potentially genotoxic metabolites, but some approaches to safety assessment are discussed for these. © 2017 Wiley Periodicals, Inc.

Sistare F.D.,Merck And Co. | Mattes W.B.,Food and Drug Administrations National Center for Toxicological Research | LeCluyse E.L.,LifeNet Health
ILAR Journal | Year: 2016

Drug induced liver injury (DILI) has contributed more to marketed pharmaceutical withdrawals and clinical development failures than any other human organ toxicity. DILI seen in animal studies also frequently leads to the discontinuation of promising drug candidates very early in the pipeline. This manuscript reviews and critically assesses the current regulatory expectations; the current drug development approaches, strategies, and gaps; and the numerous exciting opportunities becoming available to address these gaps through technological advances. Emerging integrated pharmaceutical development strategies, while far from uniform, have generally evolved to currently inform early DILI risk potential using supplemental assays for reactive metabolite formation, mitochondrial toxicity, inhibition of bile salt transport, and cellular imaging endpoints including cytotoxicity. Despite these approaches and robust animal testing, significant gaps in addressing human DILI remain. Increasingly sophisticated in vitro humanized test systems, new animal models, emerging computational models, and novel translational biomarkers are being introduced to improve our ability to more accurately predict DILI. Expectations are high for a future state with more predictive tools and problem solving strategies that will improve pharmaceutical discovery and development in relation to understanding human DILI risk potential and make it less dependent on animal studies for successfully developing safer drug candidates. © The Author 2016.

Chappell C.P.,University of Washington | Chappell C.P.,Merck And Co. | Draves K.E.,University of Washington | Clark E.A.,University of Washington
PLoS ONE | Year: 2017

CD22 is a BCR co-receptor that regulates B cell signaling, proliferation and survival and is required for T cell-independent Ab responses. To investigate the role of CD22 during T cell-dependent (TD) Ab responses and memory B cell formation, we analyzed Ag-specific B cell responses generated by wild-type (WT) or CD22-/- B cells following immunization with a TD Ag. CD22-/- B cells mounted normal early Ab responses yet failed to generate either memory B cells or long-lived plasma cells, whereas WT B cells formed both populations. Surprisingly, B cell expansion and germinal center (GC) differentiation were comparable between WT and CD22-/- B cells. CD22-/- B cells, however, were significantly less capable of generating a population of CXCR4hiCD38hi GC B cells, which we propose represent memory B cell precursors within GCs. These results demonstrate a novel role for CD22 during TD humoral responses evident during primary GC formation and underscore that CD22 functions not only during B cell maturation but also during responses to both TD and T cell-independent antigens. © 2017 Chappell et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Kinter L.B.,GLP Scientific Consulting | DeGeorge J.J.,Merck And Co.
ILAR Journal | Year: 2016

Human discovery of pharmacologically active substances is arguably the oldest of the biomedical sciences with origins > 3500 years ago. Since ancient times, four major transformations have dramatically impacted pharmaceutical development, each driven by advances in scientific knowledge, technology, and/or regulation: (1) anesthesia, analgesia, and antisepsis; (2) medicinal chemistry; (3) regulatory toxicology; and (4) targeted drug discovery. Animal experimentation in pharmaceutical development is a modern phenomenon dating from the 20th century and enabling several of the four transformations. While each transformation resulted in more effective and/or safer pharmaceuticals, overall attrition, cycle time, cost, numbers of animals used, and low probability of success for new products remain concerns, and pharmaceutical development remains a very high risk business proposition. In this manuscript we review pharmaceutical development since ancient times, describe its coevolution with animal experimentation, and attempt to predict the characteristics of future transformations. © The Author 2016.

Fier P.S.,Merck And Co. | Whittaker A.M.,Merck And Co.
Organic Letters | Year: 2017

The development of a rapid, one-pot synthesis of diazepinones with simple reagents is described. N-Carboxyanhydrides (NCAs) are employed as amino acid building blocks that react with o-ketoanilines sequentially as electrophiles and nucleophiles to form diazepinones with water and carbon dioxide as byproducts. Notably, these reactions enable the coupling of stereodefined amino acid derived NCAs without racemization. This method is demonstrated by an improved synthesis of a key intermediate toward a bromodomain and extra-terminal (BET) bromodomain inihibitor. © 2017 American Chemical Society.

Mann B.F.,Merck And Co. | Makarov A.A.,Merck And Co. | Wang H.,Merck And Co. | Welch C.J.,Merck And Co.
Journal of Chromatography A | Year: 2017

Pressure is not typically controlled or adjusted independently of flow rate during method development in reversed-phase LC (RPLC). However, it has been shown that pressure has an effect on analyte molecular molar volume, and the magnitude of this effect is greater for proteins and ionizable compounds than neutral small molecules. This phenomenon has received attention recently in the context of porous sub-2-micron particle packed columns. The present study surveys the effect of pressure and frictional heating on RPLC separations using commercially-available monolithic columns at constant flow rate and with controlled external temperature. Because the current monoliths cannot be operated at high pressures, all experiments were conducted with pressures at or below 200. bar. Nonetheless, substantial changes in retention were still observed; for example, an increase in pressure of 75. bar shifted the retention factor for bovine insulin from 1.27 to 1.78, a 40% increase, while a similar experiment with the neutral small molecule, toluene, showed no change in retention. Results are presented from investigations of model peptides and proteins ranging in size from 1. kDa to 30. kDa, as well as experiments performed with a silica-based C18 monolith and a polystyrene divinylbenzene monolith functionalized with a phenyl stationary phase. This work indicates that protein separations in monoliths are highly pressure sensitive, and pressure should therefore be considered as an additional parameter in method development for optimizing retention and selectivity. Given these findings, and the ever-increasing importance of chromatographic separations of proteins in both industrial and academic laboratories, improved instrumentation and mechanisms for directly controlling system backpressure could be of great practical value. © 2017 Elsevier B.V.

Wang F.,Merck And Co. | Ha S.,Merck And Co. | Rustandi R.R.,Merck And Co.
Journal of Chromatography A | Year: 2017

Clostridium difficile is a gram-positive intestine bacterium that causes a severe diarrhea and could eventually be lethal. The main virulence factor is related to the release of two major exotoxins, toxin A (TcdA) and toxin B (TcdB). Recent C. difficile-associated disease (CDAD) outbreaks have been caused by hypervirulent strains which secrete an additional binary toxin (CDTa/CDTb). Vaccination against these toxins is considered the best way to combat the CDAD. Recently, a novel tetravalent C. difficile vaccine candidate containing all four toxins produced from a baculovirus expression system has been developed. A dose assay to release this tetravalent C. difficile vaccine was developed using tandem ion-exchange HPLC chromatography. A sequential weak cation exchange (carboxyl group) and weak anion exchange (tertiary amine group) columns were employed. The four C. difficile vaccine antigen pIs range from 4.4 to 8.6. The final optimized separation employs salt gradient elution at two different pHs. The standard analytical parameters such as LOD, LOQ, linearity, accuracy, precision and repeatability were evaluated for this method and it was deemed acceptable as a quantitative assay for vaccine release. Furthermore, the developed method was utilized for monitoring the stability of the tetravalent C. difficile vaccine in final container. © 2017 Elsevier B.V.

Kuznetsova O.M.,Merck And Co. | Johnson V.P.,Merck And Co.
Statistics in Medicine | Year: 2017

The paper discusses three methods for expanding the biased coin randomization (BCR) to unequal allocation while preserving the unconditional allocation ratio at every step. The first method originally proposed in the contexts of BCR and minimization is based on mapping from an equal allocation multi-arm BCR. Despite the improvement proposed in this paper to ensure tighter adherence to the targeted unequal allocation, this method still distributes the probability mass at least as wide as the permuted block randomization (PBR). This works for smaller block sizes, but for larger block sizes, a tighter control of the imbalance in the treatment assignments is desired. The second method, which has two versions, allows to tighten the distribution of the imbalance compared with that achieved with the PBR. However, the distribution of the imbalance remains considerably wider than that of the brick tunnel randomization - the unequal allocation procedure with the tightest possible imbalance distribution among all allocation ratio preserving procedures with the same allocation ratio. Finally, the third method, the BCR with a preset proportion of maximal forcing, mimics the properties of the equal allocation BCR. With maximum forcing, it approaches the brick tunnel randomization, similar to how 1:1 BCR approaches 1:1 PBR with the permuted block size of 2 (the equal allocation procedure with the lowest possible imbalance) when the bias approaches 1. With minimum forcing, the BCR with a preset proportion of maximal forcing approaches complete randomization (similar to 1:1 BCR). © 2017 John Wiley & Sons, Ltd.

Sun H.,University of North Carolina at Chapel Hill | Binkowitz B.,Merck And Co. | Koch G.G.,University of North Carolina at Chapel Hill
Journal of Biopharmaceutical Statistics | Year: 2017

Multiplicity is an important statistical issue that arises in clinical trials when the efficacy of the test treatment is evaluated in multiple ways. The major concern for multiplicity is that uncontrolled multiple assessments lead to inflated family-wise Type I error, and they thereby undermine the integrity of the statistical inferences. Multiplicity comes from different sources, for example, making inferences either on the overall population or some pre-specified sub-populations, while multiple endpoints need to be evaluated for each population. Therefore, a sound statistical strategy that controls the family-wise Type I error rate in a strong sense, without excessive loss of power from over-control, is crucial for the success of the trial. For a recent phase III cardiovascular trial with such complex multiplicity, we illustrate the use of a closed testing strategy that begins with a global test; and subsequent testing only proceeds when the global test is rejected. Also, we discuss a simulation study based on this trial to compare the power of the illustrated closed testing strategy to some well-known alternative approaches. We found that this strategy can comprehensively meet most of the primary objectives of the trial effectively with reasonably high overall power. © 2017 Taylor & Francis Group, LLC

Li W.,Merck And Co. | Chen C.,Merck And Co. | Li X.,Merck And Co. | Beckman R.A.,Georgetown University
Statistics in Medicine | Year: 2017

A personalized medicine may benefit a subpopulation with certain predictive biomarker signatures or certain disease types. However, there is great uncertainty about drug activity in a subpopulation when designing a confirmatory trial in practice, and it is logical to take a two-stage approach with the study unless credible external information is available for decision-making purpose. The first stage deselects (or prunes) non-performing subpopulations at an interim analysis, and the second stage pools the remaining subpopulations in the final analysis. The endpoints used at the two stages can be different in general. A key issue of interest is the statistical property of the test statistics and point estimate at the final analysis. Previous research has focused on type I error control and power calculation for such two-stage designs. This manuscript will investigate estimation bias of the treatment effect, which is implicit in the adjustment of nominal type I error for multiplicity control in such two-stage designs. Previous work handles the treatment effect of an intermediate endpoint as a nuisance parameter to provide the most conservative type I error control. This manuscript takes the same approach to explore the bias. The methodology is applied to the two previously studied designs. In the first design, patients with different biomarker levels are enrolled in a study, and the treatment effect is assumed to be in an order. The goal of the interim analysis is to identify a biomarker cut-off point for the subpopulations. In the second design, patients with different tumour types but the same biomarker signature are included in a trial applying a basket design. The goal of the interim analysis is to identify a subset of tumour types in the absence of treatment effect ordering. Closed-form equations are provided for the estimation bias as well as the variance under the two designs. Simulations are conducted under various scenarios to validate the analytic results that demonstrated that the bias can be properly estimated in practice. Worked examples are presented. Extensions to general adaptive designs and operational considerations are discussed. © 2017 John Wiley & Sons, Ltd.

BACKGROUND: Antibacterials that inhibit protein synthesis may be associated with mitochondrial toxicity, manifested as serious optic or peripheral neuropathy or myelosuppression. Tedizolid is a novel oxazolidinone antibacterial that may have reduced the potential for mitochondrial toxicity.STUDY QUESTION: Based on the results of 2 studies (NCT01623401 and NCT00671814) conducted early in the tedizolid development program, what is the potential for drug-induced optic and peripheral neuropathies with tedizolid treatment?METHODS: Two phase-1 studies were conducted in healthy volunteers. The first was an open-label study in which subjects received 200 mg of oral tedizolid phosphate once daily for 10 days. The second was a double-blind, placebo- and active-controlled, dose-escalating (multiple-administration) study in which subjects received 200, 300, or 400 mg of oral tedizolid phosphate once daily or 600 mg of oral linezolid twice daily or oral placebo for 21 days. Overall safety and tolerability were assessed, and extensive ophthalmologic and neurologic assessments were performed in both studies.RESULTS: In these 2 studies in healthy subjects, tedizolid administered for up to 21 days was not associated with drug-related ophthalmologic or neurologic adverse events. Incidences of adverse events involving the eye or the nervous system were generally low, and no clinically meaningful changes in ophthalmologic or neurologic test results were recorded during either study.CONCLUSIONS: Using an extensive battery of ophthalmologic tests and detailed neurologic clinical examination, there was no evidence of clinical or subclinical neurologic or ophthalmologic changes suggestive of peripheral or optic neuropathy in healthy volunteers who received therapeutic and supratherapeutic doses of oral tedizolid for periods of up to 21 days.

Weyn-Vanhentenryck S.M.,Columbia University | Mele A.,Howard Hughes Medical Institute | Yan Q.,Columbia University | Sun S.,Cold Spring Harbor Laboratory | And 12 more authors.
Cell Reports | Year: 2014

The RNA binding proteins Rbfox1/2/3 regulate alternative splicing in the nervous system, and disruption of Rbfox1 has been implicated in autism. However, comprehensive identification of functional Rbfox targets has been challenging. Here, we perform HITS-CLIP for all three Rbfox family members in order to globally map, at a single-nucleotide resolution, their invivo RNA interaction sites in the mouse brain. We find that the two guanines in the Rbfox binding motif UGCAUG are critical for protein-RNA interactions and crosslinking. Using integrative modeling, these interaction sites, combined with additional datasets, define 1,059 direct Rbfox targetalternative splicing events. Over half of the quantifiable targets show dynamic changes during brain development. Of particular interest are 111 events from 48 candidate autism-susceptibility genes, including syndromic autism genes Shank3, Cacna1c, and Tsc2. Alteration of Rbfox targets in some autistic brains is correlated with downregulation of all three Rbfox proteins, supporting the potential clinical relevance of the splicing-regulatory network. © 2014 The Authors.

Masarachia P.J.,Merck And Co. | Pennypacker B.L.,Merck And Co. | Pickarski M.,Merck And Co. | Scott K.R.,Merck And Co. | And 7 more authors.
Journal of Bone and Mineral Research | Year: 2012

Odanacatib (ODN) is a selective and reversible inhibitor of cathepsin K (CatK) currently being developed as a once-weekly treatment for osteoporosis. In this study, we evaluated the effects of ODN on bone turnover, bone mineral density (BMD), and bone strength in the lumbar spine of estrogen-deficient, skeletally mature rhesus monkeys. Ovariectomized (OVX) monkeys were treated in prevention mode for 21 months with either vehicle, ODN 6 mg/kg, or ODN 30 mg/kg (p.o., q.d.) and compared with intact animals. ODN treatment persistently suppressed the bone resorption markers (urinary NTx [75% to 90%] and serum CTx [40% to 55%]) and the serum formation markers (BSAP [30% to 35%] and P1NP [60% to 70%]) versus vehicle-treated OVX monkeys. Treatment with ODN also led to dose-dependent increases in serum 1-CTP and maintained estrogen deficiency-elevated Trap-5b levels, supporting the distinct mechanism of CatK inhibition in effectively suppressing bone resorption without reducing osteoclast numbers. ODN at both doses fully prevented bone loss in lumbar vertebrae (L 1 to L 4) BMD in OVX animals, maintaining a level comparable to intact animals. ODN dose-dependently increased L 1 to L 4 BMD by 7% in the 6 mg/kg group (p < 0.05 versus OVX-vehicle) and 15% in the 30 mg/kg group (p < 0.05 versus OVX-vehicle) from baseline. Treatment also trended to increase bone strength, associated with a positive and highly significant correlation (R = 0.838) between peak load and bone mineral content of the lumbar spine. Whereas ODN reduced bone turnover parameters in trabecular bone, the number of osteoclasts was either maintained or increased in the ODN-treated groups compared with the vehicle controls. Taken together, our findings demonstrated that the long-term treatment with ODN effectively suppressed bone turnover without reducing osteoclast number and maintained normal biomechanical properties of the spine of OVX nonhuman primates. Copyright © 2012 American Society for Bone and Mineral Research.

Bigal M.E.,Merck And Co. | Kurth T.,Brigham and Women's Hospital | Santanello N.,Merck And Co. | Golden W.,Merck And Co. | And 2 more authors.
Neurology | Year: 2010

Objectives: Although the relationship between migraine and cardiovascular disease (CVD) has been studied, several questions remain unanswered. Herein we contrast the rate of diagnosed CVD as well as of risk factors for CVD in individuals with migraine with and without aura (MA and MO) and in controls. Methods: In this case-control study, migraineurs (n = 6,102) and controls (n = 5,243) were representative of the adult US population. Headache diagnosis was formally assigned using a validated mailed questionnaire which also obtained details on treatment, comorbidities, and other variables. CVD events were obtained based on self-reported medical diagnosis. Risk factors for CVD and modified Framingham scores were computed. Results: In unadjusted analyses, migraine overall and MA were associated with myocardial infarction, stroke, and claudication, and MO was associated with myocardial infarction and claudication. Migraineurs were more likely than controls to have a medical diagnosis of diabetes (12.6% vs 9.4%, odds ratio [OR] 1.4, 95% confidence interval [CI] 1.2-1.6), hypertension (33.1% vs 27.5%, OR 1.4, 95% CI 1.3-1.6), and high cholesterol (32.7% vs 25.6%, OR 1.4, 95% CI 1.3-1.5). Risk was highest in MA, but remained elevated in MO. Framingham scores were significantly higher in MO and MA than in controls. After adjustments (gender, age, disability, treatment, CVD risk factors), migraine remained significantly associated with myocardial infarction (OR 2.2, 95% CI 1.7-2.8), stroke (OR 1.5, 95% CI 1.2-2.1), and claudication (OR 2.69, 95% CI 1.98-3.23). Conclusion: Both migraine with and without aura are associated with cardiovascular disease (CVD) and with risk factors for CVD. However, since our sample size is large, the clinical relevance of the differences is yet to be established. Copyright © 2010 by AAN Enterprises, Inc. All rights reserved.

Marijnissen R.J.,Radboud University Nijmegen | Koenders M.I.,Radboud University Nijmegen | Smeets R.L.,Merck And Co. | Stappers M.H.T.,Radboud University Nijmegen | And 5 more authors.
Arthritis and Rheumatism | Year: 2011

Objective Interleukin-22 (IL-22) is a mediator in antimicrobial responses and inflammatory autoimmune diseases. Although IL-22 and its receptor, IL-22R, have been identified in the synovium of rheumatoid arthritis patients, the source of IL-22 and its contribution to disease pathogenicity remain to be established. This study was undertaken to investigate the regulation of IL-22 by Th17 cells in vitro and to evaluate the potential for IL-22 depletion in an experimental arthritis model using mice deficient in the IL-1 receptor antagonist (IL-1Ra-/-). Methods Naive murine T cells were cultured under conditions leading to polarization of the cells into subsets of Th1, Th2, induced Treg, and Th17. Cytokines were measured in the culture supernatants, and the cells were analyzed by fluorescence-activated cell sorting. Tissue samples from the inflamed ankle synovium of IL-1Ra-/- mice were isolated, and messenger RNA levels of marker genes were quantified. IL-1Ra-/- mice were treated with neutralizing anti-IL-22 antibodies. Synovial cells were isolated from the inflamed tissue and sorted into fractions for analysis of cytokine production. Results In vitro tests showed that Th17 cells produced high levels of IL-22 after stimulation with IL-1 or IL-23. Interestingly, a synergistic increase in the production of IL-22 was observed after combining IL-1 and IL-23. In vivo, IL-1Ra-/- mice displayed a progressive erosive arthritis, characterized by up-regulation of IL-17 in mildly and severely inflamed tissue, whereas the levels of IL-22 and IL-22R were increased only in severely inflamed synovia. Anti-IL-22 treatment of IL-1Ra-/- mice significantly reduced the inflammation and bone erosion. Analysis of isolated single cells from the inflamed synovia revealed that IL-22 was mainly produced by IL-17-expressing T cells. Conclusion These findings suggest that IL-22 plays an important role in IL-1-driven chronic joint destruction. Copyright © 2011 by the American College of Rheumatology.

Nie L.,OB OTS CDER FDA | Rubin E.H.,Merck And Co. | Mehrotra N.,OCP OTS CDER FDA | Pinheiro J.,Johnson and Johnson | And 4 more authors.
Clinical Cancer Research | Year: 2016

Selection of the maximum tolerated dose (MTD) as the recommended dose for registration trials based on a doseescalation trial using variations of an MTD/3 + 3 design often occurs in the development of oncology products. The MTD/3 + 3 approach is not optimal and may result in recommended doses that are unacceptably toxic for many patients and in dose reduction/interruptions that might have an impact on effectiveness. Instead of the MTD/3 + 3 approach, the authors recommend an integrated approach. In this approach, typically an adaptive/Bayesian model provides a general framework to incorporate and make decisions for dose escalation based on nonclinical data, such as animal efficacy and toxicity data; clinical data, including pharmacokinetics/pharmacodynamics data; and dose/exposure-response data for efficacy and safety. To improve dose-ranging trials, model-based estimation, rather than hypothesis testing, should be used to maximize and integrate the information gathered across trials and doses. This approach may improve identification of optimal recommended doses, which can then be confirmed in registration trials. © 2016 American Association for Cancer Research.

Shahly V.,Harvard University | Berglund P.A.,University of Michigan | Coulouvrat C.,Sanofi S.A. | Fitzgerald T.,Merck And Co. | And 6 more authors.
Archives of General Psychiatry | Year: 2012

Context: Insomnia is a common and seriously impairing condition that often goes unrecognized. Objectives: To examine associations of broadly defined insomnia (ie, meeting inclusion criteria for a diagnosis from International Statistical Classification of Diseases, 10th Revision , DSM-IV , or Research Diagnostic Criteria/International Classification of Sleep Disorders, Second Edition) with costly workplace accidents and errors after excluding other chronic conditions among workers in the America Insomnia Survey (AIS). Design/Setting: A national cross-sectional telephone survey (65.0% cooperation rate) of commercially insured health plan members selected from the more than 34 million in the HealthCore Integrated Research Database. Participants : Four thousand nine hundred ninetyone employed AIS respondents. Main Outcome Measures: Costly workplace accidents or errors in the 12 months before the AIS interview were assessed with one question about workplace accidents "that either caused damage or work disruption with a value of $500 or more" and another about other mistakes "that cost your company $500 or more." Results: Current insomnia with duration of at least 12 months was assessed with the Brief Insomnia Questionnaire, a validated (area under the receiver operating characteristic curve, 0.86 compared with diagnoses based on blinded clinical reappraisal interviews), fully structured diagnostic interview. Eighteen other chronic conditions were assessed with medical/pharmacy claims records and validated self-report scales. Insomnia had a significant odds ratio with workplace accidents and/or errors controlled for other chronic conditions (1.4). The odds ratio did not vary significantly with respondent age, sex, educational level, or comorbidity. The average costs of insomnia-related accidents and errors ($32 062) were significantly higher than those of other accidents and errors ($21 914). Simulations estimated that insomnia was associated with 7.2% of all costly workplace accidents and errors and 23.7% of all the costs of these incidents. These proportions are higher than for any other chronic condition, with annualized US population projections of 274 000 costly insomnia-related workplace accidents and errors having a combined value of US $31.1 billion. Conclusion: Effectiveness trials are needed to determine whether expanded screening, outreach, and treatment of workers with insomnia would yield a positive return on investment for employers. ©2012 American Medical Association. All rights reserved.

Marsico M.,Merck And Co. | Mehta V.,Merck And Co. | Liaw K.-L.,Merck And Co. | Derkay C.,Eastern Virginia Medical School
Sexually Transmitted Diseases | Year: 2014

BACKGROUND: Juvenile-onset recurrent respiratory papillomatosis (JORRP) is a chronic disease caused by human papillomavirus types 6 and 11. It is associated with significant morbidity that places intense physical, psychological, and financial strain on patients and their families. Few studies have assessed the incidence and prevalence of JORRP in the United States. METHODS: This retrospective, longitudinal cohort study was performed using data from a pair of large insurance claims databases in the United States. The Optum Clinformatics and Truven MarketScan Medicaid databases represent a sample of privately and publicly insured children, respectively. Cohorts of children aged 0 to 17 years were created within each database to estimate the incidence and prevalence of JORRP in 2006. Claims-based algorithms were designed to capture as many potential cases as possible. To improve the accuracy of the incidence and prevalence estimates, chart validation was performed to estimate the positive predictive value (PPV) of the claims-based algorithms. RESULTS: The overall PPV-adjusted incidence of JORRP in 2006 was 0.51 per 100,000 in Optum and 1.03 per 100,000 in the MarketScan Medicaid population. Peak incidence was observed among 0-to 4-year-olds in both databases. The PPV-adjusted prevalence of JORRP in 2006 was 1.45 and 2.93 per 100,000 in the Optum and MarketScan Medicaid cohorts, respectively. CONCLUSIONS: Although relatively uncommon, JORRP represents a disease with significant morbidity. The incidence and prevalence of JORRP in publicly insured children were consistently higher than those covered by private insurance plans, suggesting an increased burden of illness among those with lower socioeconomic status. © 2014 by the American Sexually Transmitted Diseases Association.

Klimas M.,Merck And Co.
JACC: Cardiovascular Imaging | Year: 2013

Objectives This study hypothesized that 1H magnetic resonance spectroscopy (1H-MRS) can identify carotid plaque cholesteryl ester in vivo in humans. Background Liquid phase cholesteryl ester comprises a major fraction of atherosclerotic plaque, and its abundance is associated with plaque rupture and atherothrombosis. A noninvasive imaging technique to detect liquid cholesteryl ester that has been applied ex vivo is now demonstrated in vivo. Methods 1H-MRS scans were obtained of carotid plaques of 35 subjects at 3.0 T. Turbo spin echo, black blood, T1-weighted images were acquired for localization. Spectra were acquired using a 2-dimensional point resolved spectroscopy sequence: repetition time/echo time = 1,100/30 ms, 5-mm slice thickness, 8 × 8-cm field of view, 16 × 16 matrix size, and 13-min acquisition time. Saturation bands were placed around the artery. Resonance of methylene protons and allylic methylene protons were assigned to 1.2 ppm and 2.0 ppm. The 2.0:1.2 ppm ratio was calculated to reflect the ratio of the fatty acid composition of plaque cholesteryl ester to that of triglycerides of perivascular tissue. We obtained spectra of lipid standards as a reference. Results Our 1H-MRS data showed typical spectra of cholesteryl ester mixed with triglycerides, with intense resonance from methylene (1.2 ppm) and allylic methylene (2.0 ppm) protons. The average 2.0:1.2 ppm ratio was 0.10 ± 0.03. The 2.0:1.2 ppm ratio correlated with the plaque tissue volume to perivascular tissue volume ratio (Spearman rho = 0.55, p = 0.02), suggesting that more 1H-MRS signal was obtained from cholesteryl ester when the 1H-MRS voxel comprised more plaque tissue. Repeat 1H-MRS scans in 4 subjects showed an intraclass correlation coefficient of 0.92 (95% prediction intervals: 0.40 to 0.99), indicating good reproducibility. Seventeen of the 35 1H-MRS spectra were of adequate quality for analysis. Conclusions In vivo image-guided 1H-MRS for detection of liquid phase cholesteryl ester in carotid atherosclerotic plaques in humans is feasible. © 2013 by the American College of Cardiology Foundation Published by Elsevierinc .

Lane N.E.,University of California at Davis | Brandt K.,University of Kansas Medical Center | Hawker G.,University of Toronto | Peeva E.,Merck And Co. | And 3 more authors.
Osteoarthritis and Cartilage | Year: 2011

Objective: To respond to a pre-specified set of questions posed by the United States Food and Drug Administration (FDA) on defining the disease state to inform the clinical development of drugs, biological products, and medical devices for the prevention and treatment of osteoarthritis (OA). Methods: An Osteoarthritis Research Society International (OARSI) Disease State working group was established, comprised of representatives from academia and industry. The Working Group met in person and by teleconference on several occasions from the Spring of 2008 through the Autumn of 2009 to develop consensus-based, evidence-informed responses to these questions. A report was presented at a public forum in December 2009 and accepted by the OARSI Board of Directors in the Summer of 2010. Results: An operational definition of OA was developed incorporating current understanding of the condition. The structural changes that characterize OA at the joint level were distinguished from the patients' experience of OA as the 'disease' and 'illness', respectively. Recommendations were made regarding the evaluation of both in future OA clinical trials. The current poor understanding of the phenotypes that characterize OA was identified as an important area for future research. Conclusions: The design and conduct of clinical trials for new OA treatments should address the heterogeneity of the disease, treatment-associated structural changes in target joints and patient-reported outcomes. © 2011 Osteoarthritis Research Society International.

Okin P.M.,Cornell University | Bang C.N.,Cornell University | Wachtell K.,Gentofte Hospital | Hille D.A.,Merck And Co. | And 4 more authors.
Circulation: Arrhythmia and Electrophysiology | Year: 2013

Background-Prevalent atrial fibrillation (AF) is associated with a higher sudden cardiac death (SCD) rate in some populations, and incident AF predicts increased mortality risk in the general population and after myocardial infarction. However, the relationship of SCD to new-onset AF is unclear. Methods and Results-The relationship of SCD to new-onset AF was evaluated in 8831 hypertensive patients with electrocardiographic left ventricular hypertrophy with no history of AF, in sinus rhythm on their baseline electrocardiogram, randomly assigned to losartan- or atenolol-based treatment. During 4.7±1.1 years mean follow-up, new-onset AF occurred in 701 patients (7.9%) and SCD in 151 patients (1.7%). In univariate Cox analyses, new-onset AF was associated with a > 4-fold higher risk of SCD (hazard ratio, 4.69; 95% CI interval, 2.96-7.45; P<0.001). In multivariate Cox analyses adjusting for age, sex, race, diabetes mellitus, history of heart failure, myocardial infarction, ischemic heart disease, stroke, smoking, serum high-density lipoprotein cholesterol, creatinine, glucose, and urine albumin/creatinine ratio as standard risk factors, and for incident myocardial infarction, in-treatment use of digoxin, systolic and diastolic pressure, heart rate, QRS duration, Cornell voltage-duration product, and Sokolow-Lyon voltage left ventricular hypertrophy treated as time-varying covariates, new-onset AF remained associated with a > 3-fold increased risk of SCD (hazard ratio, 3.13; 95% confidence interval, 1.87-5.24; P<0.001). Conclusions-Development of new-onset AF identifies hypertensive patients at increased risk of SCD. © 2013 American Heart Association, Inc.

Rolan P.,University of Adelaide | Sun H.,Novartis | MacLeod C.,Merck And Co. | Bracken K.,Novartis | Evans T.G.,Novartis
Clinical Pharmacology and Therapeutics | Year: 2011

Peptide deformylase (PDF) inhibitors represent a potential new class of antibiotics targeting a large number of bacterial species. We studied the pharmacokinetics and safety of LBM415, a novel PDF inhibitor, administered as a single oral dose at 100-3,000mg in the fasted state and at 1,000mg in the fed state in healthy volunteers. LBM415 was then administered at dosages ranging from 100mg q.d. to 1,000mg t.i.d. for 11 days. Dose-proportional pharmacokinetics was observed, with a peak plasma concentration (C max) of 17.85 5.96νg/ml at 1,000mg b.i.d. (the projected therapeutic dose) and an area under the concentration-time curve (AUC) 0-24h of 36.83 10.36νg/mlh. The half-life, as determined after a 1,000-mg single dose, was 2.18 0.61h. The compound was well tolerated at low doses, but at the highest dose, 1,000mg t.i.d., reversible cyanosis and low oxygen saturation, attributable to methemoglobinemia, were detected on day 11. Oxygen saturation was as low as 88% in one subject on day 11. © 2011 american society for Clinical Pharmacology and therapeutics.

Okin P.M.,Cornell University | Kjeldsen S.E.,University of Oslo | Kjeldsen S.E.,University of Michigan | Julius S.,University of Michigan | And 4 more authors.
European Heart Journal | Year: 2010

BackgroundAlthough higher heart rate (HR) at baseline has been associated with an increased risk of cardiovascular (CV) and all-cause mortality, the relationship of in-treatment HR over time to mortality in hypertensive patients with ECG left ventricular hypertrophy (LVH) has not been examined. Methods and resultsHeart rate was evaluated over time in 9190 hypertensive patients treated with losartan- or atenolol-based regimens and followed with annual ECGs. During a mean follow-up of 4.8 ± 0.9 years, 814 patients (8.9) died, 438 (4.8) from CV causes. In univariate Cox analyses, every 10 bpm higher HR on in-treatment ECGs was associated with a 25 increased risk of CV death [95 confidence interval (CI): 14-32] and a 27 greater risk of all-cause mortality (95 CI: 21-34). In an alternative analysis, persistence or development of a HR ≥84 bpm (upper quintile of baseline HR) was associated with an 89 greater risk of CV death (95 CI: 49-141) and a 97 increased risk of all-cause mortality (95 CI: 65-135). After adjusting for treatment with losartan vs. atenolol, baseline risk factors for death, baseline HR, baseline and in-treatment systolic and diastolic pressure, incident myocardial infarction, and the known predictive value of baseline and in-treatment QRS duration and ECG LVH, higher in-treatment HR in time-varying multivariable Cox models remained strongly predictive of mortality: every 10 bpm higher HR was associated with a 16 increased adjusted risk of CV mortality (95 CI: 6-27) and a 25 greater risk of all-cause mortality (95 CI: 17-33), with persistence or development of a HR ≥84 associated with a 55 greater risk of CV death (95 CI: 16-105) and a 79 greater adjusted risk of all-cause mortality (95 CI: 46-121). ConclusionHigher in-treatment HR on serial ECGs predicts greater likelihood of subsequent CV or all-cause mortality, independent of treatment modality, blood pressure lowering, regression of ECG LVH and changing QRS duration in hypertensive patients with ECG LVH. These findings support the value of serial assessment of HR for improved risk stratification in hypertensive patients. © 2010 The Author.

Van Der Meer P.,University of Groningen | Postmus D.,University of Groningen | Ponikowski P.,Klinika Kardiologii | Cleland J.G.,University of Hull | And 10 more authors.
Journal of the American College of Cardiology | Year: 2013

Objectives: The study sought to investigate the clinical correlates and prognostic role of anemia and changes in hemoglobin in patients hospitalized for acute decompensated heart failure (AHF). Background: Anemia is related to a poor outcome in patients with heart failure. In addition, an increase in hemoglobin during hospitalization might be a sign of effective decongestion and therefore related to improved outcome. Methods: This is a post hoc analysis of the PROTECT (Placebo-Controlled Randomized Study of the Selective Adenosine A1 Receptor Antagonist Rolofylline for Patients Hospitalized with Acute Decompensated Heart Failure and Volume Overload to Assess Treatment Effect on Congestion and Renal Function) study in 1,969 patients with AHF and mild to moderate impaired renal function. Hemoglobin levels were measured daily for the first 4 days and at day 7. The endpoint was 180-day all-cause mortality. Results: Anemia at baseline was observed in 50.3% of the patients. During follow-up, 359 patients (18.2%) died. Hemoglobin increased in 69.1% and was associated with a better renal function at baseline and more weight loss, but was associated with a deterioration of renal function (p = 0.01), whereas total dose diuretics was lower in patients with hemoconcentration (p < 0.01). Interaction analysis showed that greater weight loss and better baseline renal function were associated with a more rapid increase in hemoglobin concentration (p < 0.01 for both). The absolute change in hemoglobin (g/dl) independently predicted outcome (hazard ratio: 0.66; 95% confidence interval: 0.51 to 0.86; p = 0.002), whereas baseline hemoglobin levels did not. Conclusions: Patients with AHF and preserved renal function are decongested better, as shown by an increase in hemoglobin. A rapid increase in hemoglobin during the first week is independently associated with a favorable outcome, despite a slight decrease in renal function. © 2013 American College of Cardiology Foundation.

Kudryashova E.,University of California at Los Angeles | Struyk A.,Merck And Co. | Mokhonova E.,University of California at Los Angeles | Spencer M.J.,University of California at Los Angeles
Human Molecular Genetics | Year: 2011

Mutations in tripartite motif protein 32 (TRIM32) are responsible for several hereditary disorders that include limb girdle muscular dystrophy type 2H (LGMD2H), sarcotubular myopathy (STM) and Bardet Biedl syndrome. Most LGMD2H mutations in TRIM32 are clustered in the NHL β-propeller domain at the C-terminus and are predicted to interfere with homodimerization. To get insight into TRIM32's role in the pathogenesis of LGMD2H and to create an accurate model of disease, we have generated a knock-in mouse (T32KI) carrying the c.1465G > A (p.D489N) mutation in murine Trim32 corresponding to the human LGMD2H/STM pathogenic mutation c.1459G > A (p.D487N). Our data indicate that T32KI mice have both a myopathic and a neurogenic phenotype, very similar to the one described in the Trim32-null mice that we created previously. Analysis of Trim32 gene expression in T32KI mice revealed normal mRNA levels, but a severe reduction in mutant TRIM32 (D489N) at the protein level. Our results suggest that the D489N pathogenic mutation destabilizes the protein, leading to its degradation, and results in the same mild myopathic and neurogenic phenotype as that found in Trim32-null mice. Thus, one potential mechanism of LGMD2H might be destabilization of mutated TRIM32 protein leading to a null phenotype. © The Author 2011. Published by Oxford University Press. All rights reserved.

Lontok E.,University of California at Berkeley | Harrington P.,U.S. Food and Drug Administration | Howe A.,Merck And Co. | Kieffer T.,Vertex Pharmaceuticals | And 7 more authors.
Hepatology | Year: 2015

Hepatitis C virus (HCV) drug development has resulted in treatment regimens composed of interferon-free, all-oral combinations of direct-acting antivirals. While the new regimens are potent and highly efficacious, the full clinical impact of HCV drug resistance, its implications for retreatment, and the potential role of baseline resistance testing remain critical research and clinical questions. In this report, we discuss the viral proteins targeted by HCV direct-acting antivirals and summarize clinically relevant resistance data for compounds that have been approved or are currently in phase 3 clinical trials. Conclusion: This report provides a comprehensive, systematic review of all resistance information available from sponsors' trials as a tool to inform the HCV drug development field. © 2015 by the American Association for the Study of Liver Diseases.

Schoepp D.,Merck And Co. | Kalivas P.W.,Medical University of South Carolina | Volkow N.D.,U.S. National Institute on Drug Abuse | Zarate C.,National Health Research Institute | And 6 more authors.
Science Translational Medicine | Year: 2011

The neurotransmitter glutamate is the primary excitatory neurotransmitter in mammalian brain and is responsible for most corticocortical and corticofugal neurotransmission. Disturbances in glutamatergic function have been implicated in the pathophysiology of several neuropsychiatric disorders-including schizophrenia, drug abuse and addiction, autism, and depression-that were until recently poorly understood. Nevertheless, improvements in basic information regarding these disorders have yet to translate into Food and Drug Administration-approved treatments. Barriers to translation include the need not only for improved compounds but also for improved biomarkers sensitive to both structural and functional target engagement and for improved translational models. Overcoming these barriers will require unique collaborative arrangements between pharma, government, and academia. Here, we review a recent Institute of Medicine-sponsored meeting, highlighting advances in glutamatergic theories of neuropsychiatric illness as well as remaining barriers to treatment development.

Lockley W.J.S.,University of Surrey | Hesk D.,Merck And Co.
Journal of Labelled Compounds and Radiopharmaceuticals | Year: 2010

Metal-catalysed hydrogen isotope exchange has been widely used in the preparation of deuterium- and tritium-labelled compounds. This review details the development and utility of homogeneous and heterogeneous rhodium and ruthenium catalysts for the preparation of deuterium- and tritium-labelled compounds by hydrogen isotope exchange methodology. Copyright © 2010 John Wiley & Sons, Ltd.

Nti-Gyabaah J.,Merck And Co. | Gbewonyo K.,BioResources International Inc. | Chiew Y.C.,Rutgers University
Journal of Chemical and Engineering Data | Year: 2010

The solubility of artemisnin in 12 different organic solvents, methanol, ethanol, butanol, acetone, ethyl acetate, isopropyl acetate, acetonitrile, hexane, heptane, 2-butanone (MEK), methyl tert-butyl ether (MTBE), and toluene, as well as in three binary solvent mixtures of ethyl acetate + ethanol, ethyl acetate + acetone, and ethanol + acetone, within the temperature range of (284.10 and 323.15) K, is obtained. The solubility data were fitted to the Two-Liquid-Non-Random (NRTL) activity coefficient equation to obtain the model binary interaction parameters, which were used to predict the solubility of artemisinin in ethyl acetate and acetone or ethanol binary solvent mixtures. The predicted values compared favorably with the experimental data. © 2010 American Chemical Society.

De Villiers T.,University of Cape Town | Odio A.,Heritage Medical Research Institute | Palacios S.,Instituto Palacios | Chapurlat R.,Institute National Of La Sante | And 5 more authors.
Journal of Clinical Endocrinology and Metabolism | Year: 2013

Context: Odanacatib (ODN) is a selective cathepsin K inhibitor being developed to treat osteoporosis. Objective: The effects ofODNwere evaluatedonbonemineral density (BMD), biochemical markers of bone turnover, and safety in patients previously treated with alendronate. Design: This was a randomized, double-blind, placebo-controlled, 24-month study. Setting: The study was conducted at private or institutional practices. Participants: Postmenopausal women (n = 243) ≥60 years of age with low BMD at the total hip, femoral neck, or trochanter (T-score≤-2.5 but>-3.5 without prior fracture or≤-1.5 but>-3.5 with prior fracture) on alendronate for ≥3 years. Intervention: The intervention included ODN 50 mg or placebo weekly. Main Outcome Measures: The primary end point was percentage change from baseline of femoralneck BMD at month 24. BMD was assessed by dual-energy x-ray absorptiometry at baseline and 6, 12, and 24 months. Biochemical markers of bone turnover (serum C-telopeptides of type 1 collagen,urinary N-telopeptides of type 1 collagen, serum bone specific alkaline phosphatase, and serum N-terminal propeptide of type 1 collagen) were measured at baseline and 3, 6, 12, 18, and 24 months. Results: In the ODN group, BMD changes from baseline at the femoral neck, trochanter, total hip, and lumbar spine at 24 months (1.7%, 1.8%, 0.8%, and 2.3%, respectively) were significantly different from the placebo group. ODN significantly decreased urinary N-telopeptides of type 1collagen to creatinine ratio and significantly increased serum N-terminal propeptide of type 1 collagen compared with placebo. Serum C-telopeptides of type 1 collagen was unexpectedly increasedwith ODN treatment. The safety profile appeared similar between groups.Conclusions: ODN provided incremental BMD gains in osteoporotic women after alendronate treatment. © 2013 by The Endocrine Society.

Burashnikov A.,Masonic Medical Research Laboratory | Pourrier M.,Cardiome Pharma | Gibson J.K.,AAKVSL Pharma Consulting LLC. | Lynch J.J.,Merck And Co. | Antzelevitch C.,Masonic Medical Research Laboratory
Circulation: Arrhythmia and Electrophysiology | Year: 2012

Background-Several clinical trials have shown that vernakalant is effective in terminating recent onset atrial fibrillation (AF). The electrophysiological actions of vernakalant are not fully understood. Methods and Results-Here we report the results of a blinded study comparing the in vitro canine atrial electrophysiological effects of vernakalant, ranolazine, and dl-sotalol. Action potential durations (APD 50,75,90), effective refractory period (ERP), post repolarization refractoriness (PRR), maximum rate of rise of the action potential (AP) upstroke (V max), diastolic threshold of excitation (DTE), conduction time (CT), and the shortest S1-S1 permitting 1:1 activation (S1-S1) were measured using standard stimulation and microelectrode recording techniques in isolated normal, non-remodeled canine arterially perfused left atrial preparations. Vernakalant caused variable but slight prolongation of APD 90 (P=not significant), but significant prolongation of APD 50 at 30 μmol/L and rapid rates. In contrast, ranolazine and dl-sotalol produced consistent concentration- and reverse rate-dependent prolongation of APD 90. Vernakalant and ranolazine caused rate-dependent, whereas dl-sotalol caused reverse rate-dependent, prolongation of ERP. Significant rate-dependent PRR developed with vernakalant and ranolazine, but not with dl-sotalol. Other sodium channel-mediated parameters (ie, V max, CT, DTE, and S1-S1) also were depressed significantly by vernakalant and ranolazine, but not by dl-sotalol. Only vernakalant elevated AP plateau voltage, consistent with blockade of ultrarapid delayed rectified potassium current and transient outward potassium current. Conclusions-In isolated canine left atria, the effects of vernakalant and ranolazine were characterized by use-dependent inhibition of sodium channel-mediated parameters, and those of dl-sotalol by reverse rate-dependent prolongation of APD 90 and ERP. This suggests that during the rapid activation rates of AF, the INa blocking action of the mixed ion channel blocker vernakalant takes prominence. This mechanism may explain vernakalant's anti-AF efficacy. © 2012 American Heart Association, Inc.

Burlina F.,University Pierre and Marie Curie | Morris C.,UK National Institute for Medical Research | Behrendt R.,Merck and Cie | White P.,Merck And Co. | Offer J.,UK National Institute for Medical Research
Chemical Communications | Year: 2012

We report a simplified procedure for the chemical ligation of peptides by using the sulfamylbutyryl linker as a mildly activating group capable of participating in ligation. When the peptidyl N-methylsulfonamide is directly added with excess thiols to ligation reactions, the speed of reaction is comparable to native chemical ligation. This journal is © The Royal Society of Chemistry 2012.

Mittra E.S.,Molecular Imaging Program | Fan-Minogue H.,Molecular Imaging Program | Lin F.I.,Molecular Imaging Program | Sriram V.,Merck And Co. | And 3 more authors.
Clinical Cancer Research | Year: 2013

Purpose: Ficlatuzumab is a novel therapeutic agent targeting the hepatocyte growth factor (HGF)/c-MET pathway. We summarize extensive preclinical work using this agent in a mouse brain orthotopic model of glioblastoma. Experimental Design: Sequential experiments were done using eight- to nine-week-old nude mice injected with 3 × 105 U87 MG (glioblastoma) cells into the brain. Evaluation of ficlatuzumab dose response for this brain tumor model and comparison of its response to ficlatuzumab and to temozolamide were conducted first. Subsequently, various small-animal imaging modalities, including bioluminescence imaging (BLI), positron emission tomography (PET), and MRI, were used with a U87 MG-Luc 2 stable cell line, with and without the use of ficlatuzumab, to evaluate the ability to noninvasively assess tumor growth and response to therapy. ANOVA was conducted to evaluate for significant differences in the response. Results: There was a survival benefit with ficlatuzumab alone or in combination with temozolamide. BLI was more sensitive than PET in detecting tumor cells. Fluoro-D-thymidine (FLT) PET provided a better signal-to-background ratio than 2[18F]fluoro-2-deoxy-D-glucose (FDG) PET. In addition, both BLI and FLT PET showed significant changes over time in the control group as well as with response to therapy. MRI does not disclose any time-dependent change. Also, the MRI results showed a temporal delay in comparison to the BLI and FLT PET findings, showing similar results one drug cycle later. Conclusions: Targeting the HGF/c-MET pathway with the novel agent ficlatuzumab appears promising for the treatment of glioblastoma. Various clinically applicable imaging modalities including FLT, PET, and MRI provide reliable ways of assessing tumor growth and response to therapy. Given the clinical applicability of these findings, future studies on patients with glioblastoma may be appropriate. © 2013 AACR.

Brod M.,Julia Group, The | Fennema H.,Merck And Co.
Health and Quality of Life Outcomes | Year: 2013

Background: Controlled Ovarian Stimulation (COS) is the first step for in vitro fertilization (IVF) treatment, a treatment often described and experienced as stressful to patients and their partners. COS also requires concerted efforts by the patients in administering medication and general compliance to treatment protocols. Little is known about the impacts on patients that may be specific to this important first step in treatment. The absence of a conceptually sound and well-validated measure assessing patient experience and functioning during ovarian stimulation has been an obstacle to understanding the impacts of ovarian stimulation on women pursuing IVF. To address this gap, the Controlled Ovarian Stimulation Impact Measure (COSI) was developed based upon accepted methods for designing patient reported outcome (PRO) measures. The purpose of this study was to psychometrically validate the COSI.Methods: 267 patients from three countries (Ireland, United Kingdom, United States) were administered the COSI. Psychometric validation was conducted according to an a priori statistical analysis plan.Results: The final 28-item COSI was found to have robust scale structure with four domains: Interference in Daily Life (Work and Home), Injection Burden, Psychological Health and Compliance Worry. Internal consistency of all domains was adequate (between 0.80 to 0.87) as was test-retest reliability (between 0.72-0.87). All a-priori hypotheses for convergent and known-groups validity tests were met.Conclusions: There is a measurable impact of COS on patient functioning and well-being. The COSI is a well-developed and validated PRO measure of this impact. Future work should include examination of responsiveness and confirmation of concepts in non-western countries. © 2013 Brod and Fennema; licensee BioMed Central Ltd.

Arruda M.A.,Glia Institute | Guidetti V.,University of Rome La Sapienza | Galli F.,University of Rome La Sapienza | Bigal M.E.,Merck And Co. | Bigal M.E.,Yeshiva University
Neurology | Year: 2010

Objectives: To estimate the prevalence of chronic daily headaches (CDH) and of high-frequency episodic headaches (HFEH) in preadolescent children from the general population. Background: Early-onset cases of neurologic diseases often reflect increased biologic predisposition, specific risk factors, or both. Methods: Of 2,173 children identified as the target sample, consents were obtained from 1,870 (86.0%), and analyzable data were provided by 1,547 (71.2%). Parents and children were interviewed using a questionnaire consisting of 97 questions, with a validated headache module (10 questions). Crude and adjusted prevalences of HFEH (10-14 headache days per month) and CDH (15 or more headache days per month) were calculated. Results: The prevalence of CDH was 1.68% (girls 2.09%, boys 1.33%). The overall prevalence of HFEH was 2.52% (girls 2.8%, boys 2.3%). After adjusting for gender, age, parental history of headaches, income, and school of origin, the prevalence of CDH was higher in girls than in boys (2.2% vs 1.1%, p < 0.01) and in nonwhite vs white children (2.2% vs 1.2%, p < 0.01). Similar differences were seen for HFEH (girls 3.1%, boys 2.0%, p < 0.01), (nonwhite 3.1%, white 1.9%, p < 0.01). Income significantly contributed to the model. Conclusion: High-frequency episodic headaches and chronic daily headaches are common in the preadolescent pediatric population. Health care providers and educators should be aware of the magnitude of the problem to properly identify and treat children with headaches. Copyright © by AAN Enterpnsos, Inc.

Mozley P.D.,Merck And Co. | Schwartz L.H.,Columbia University | Bendtsen C.,Merck And Co. | Zhao B.,Columbia University | And 2 more authors.
Annals of Oncology | Year: 2010

Specific Aim: To review the evidence indicating that volumetric image analysis of computed tomography scans meets specifications for qualification as a biomarker in clinical trials or the management of individual patients with lung cancer. Methods: Claims of value were broken down into questions about technical feasibility, accuracy, the precision of measurement, sensitivity, the correlations with health outcomes, and the risks of producing misleading information. For each claim, the pertinent literature was reviewed. Results: Technical feasibility has now been shown, but only in limited contexts. Accuracy has been demonstrated, but only for tumors with favorable anatomical features. Measurement error still makes the assessment of change in small nodules precarious in diagnostic settings unless rigorous image acquisition and analysis procedures are followed. Precision is sufficient in some larger masses to make volumetrics a sensitive biomarker. In a few trials, correlations with clinical outcomes have been higher for volumetric-based measures than for unidimensional or bidimensional diameters. Value in clinical practice settings and clinical trials has been suggested, but not proven. Conclusion: The weight of the evidence indicates there are circumstances in which volumetric image analysis adds value to clinical trial science and the practice of medicine. © The Author 2010. Published by Oxford University Press on behalf of the European Society for Medical Oncology. All rights reserved.

Voors A.A.,University of Groningen | Dittrich H.C.,NovaCardia Inc. | Massie B.M.,University of California at San Francisco | Delucca P.,Merck And Co. | And 10 more authors.
Journal of the American College of Cardiology | Year: 2011

Objectives: This study sought to assess the effects of rolofylline on renal function in patients with acute heart failure (AHF) and renal dysfunction randomized in PROTECT (Placebo-Controlled Randomized Study of the Selective A1 Adenosine Receptor Antagonist Rolofylline for Patients Hospitalized With Acute Decompensated Heart Failure and Volume Overload to Assess Treatment Effect on Congestion and Renal Function). Background: Small studies have indicated that adenosine A1 receptor antagonists enhance diuresis and may improve renal function in patients with chronic heart failure or AHF. Methods: A total of 2,033 patients with AHF, volume overload, estimated creatinine clearance between 20 and 80 ml/min, and elevated natriuretic peptide levels were randomized (2:1) within 24 h of hospital presentation to rolofylline 30 mg/day or intravenous placebo for up to 3 days. Creatinine was measured daily until discharge or day 7 and on day 14. Persistent worsening renal function was defined as an increase in serum creatinine <0.3 mg/dl at both days 7 and 14, or initiation of hemofiltration or dialysis or death by day 7. Results: At baseline, mean ± SD estimated creatinine clearance was 51.0 ± 20.5 ml/min in the placebo group and 50.4 ± 20.0 ml/min in the rolofylline group. Changes in creatinine and estimated creatinine clearance were similar between placebo- and rolofylline-treated patients during hospitalization and at day 14. After 4 days, mean body weight was reduced by 2.6 and 3.0 kg in placebo and rolofylline patients, respectively (p = 0.005). Persistent worsening renal function occurred in 13.7% of the placebo group and 15.0% of the rolofylline group (odds ratio vs. placebo: 1.11 [95% confidence interval: 0.85 to 1.46]; p = 0.44). Conclusions: In this large, phase III clinical trial, the adenosine A1 receptor antagonist rolofylline did not prevent persistent worsening renal function in AHF patients with volume overload and renal dysfunction. (A Study of the Selective A1 Adenosine Receptor Antagonist KW-3902 for Patients Hospitalized With Acute HF and Volume Overload to Assess Treatment Effect on Congestion and Renal Function [PROTECT-1], NCT00328692; and [PROTECT-2], NCT00354458) © 2011 American College of Cardiology Foundation.

Okin P.M.,New York Medical College | Hille D.A.,Merck And Co. | Kjeldsen S.E.,University of Oslo | Kjeldsen S.E.,University of Michigan | And 2 more authors.
Journal of Hypertension | Year: 2012

Background: Hypertensive patients with ECG left-ventricular hypertrophy (LVH) are at increased risk of cardiovascular morbidity and mortality, and regression of ECG LVH is associated with improved cardiovascular outcomes. Although tighter control of systolic blood pressure (SBP) has been associated with a lower rate of ECG LVH, whether tighter vs. standard control of SBP is associated with greater reduction of cardiovascular risk is unclear. Methods and Results: Risk of stroke, myocardial infarction (MI), cardiovascular death, the composite endpoint of these events and all-cause mortality were examined in relation to in-treatment achieved SBP in 9193 hypertensive patients with ECG LVH randomly assigned to losartan or atenolol-based treatment. Patients with in-treatment SBP 130 mmHg or less (lowest quintile at last measurement) and SBP between 131 and 141 mmHg were compared with patients with in-treatment SBP at least 142 mmHg (median SBP at last measurement). In univariate analyses, compared with in-treatment SBP at least 142 mmHg, in-treatment SBP between 131 and 141 mmHg entered as a time-varying covariate identified patients with significantly lower risk of all events. In contrast, patients with SBP 130 mmHg or less had less reduction in MI, stroke and composite endpoint and no significant decrease in cardiovascular or all-cause mortality. In multivariate Cox analyses adjusting for baseline risk factors and randomized treatment as standard covariates and in-treatment diastolic BP, heart rate and Cornell product LVH as time-varying covariates, an in-treatment achieved SBP of 131 to 141 mmHg remained associated with a significantly decreased risk of MI, stroke and the LIFE composite endpoint. In contrast, patients who achieved a SBP 130 mmHg or less had no significant reduction in risk of MI, stroke or composite endpoint, had a trend to increased cardiovascular mortality [hazard ratio 1.32, 95% confidence interval (CI) 0.97-1.81, P = 0.078] and a statistically significant 37% increased risk of death from any cause (hazard ratio 1.37, 95% CI 1.10-1.71, P = 0.005). Conclusions: Achieved SBP 130 mmHg or less is not associated with lower cardiovascular risk than SBP of 131 to 141 mmHg and is associated with a significantly increased risk of death and trend towards increased cardiovascular mortality. These findings support the need for randomized evaluation of treatment to more aggressive vs. conventional SBP targets. © 2012 Wolters Kluwer Health | Lippincott Williams & Wilkins.

Eron J.J.,University of North Carolina at Chapel Hill | Cooper D.A.,University of New South Wales | Steigbigel R.T.,State University of New York at Stony Brook | Clotet B.,Hospital Germans Trias i Pujol | And 14 more authors.
The Lancet Infectious Diseases | Year: 2013

Background: Two randomised, placebo-controlled trials-BENCHMRK-1 and BENCHMRK-2-investigated the efficacy and safety of raltegravir, an HIV-1 integrase strand-transfer inhibitor. We report final results of BENCHMRK-1 and BENCHMRK-2 combined at 3 years (the end of the double-blind phase) and 5 years (the end of the study). Methods: Integrase-inhibitor-naive patients with HIV resistant to three classes of drug and who were failing antiretroviral therapy were enrolled. Patients were randomly assigned (2:1) to raltegravir 400 mg twice daily or placebo, both with optimised background treatment. Patients and investigators were masked to treatment allocation until week 156, after which all patients were offered open-label raltegravir until week 240. The primary endpoint was previously assessed at 16 weeks. We assessed long-term efficacy with endpoints of the proportion of patients with an HIV viral load of less than 50 copies per mL and less than 400 copies per mL, and mean change in CD4 cell count, at weeks 156 and 240. Findings: 1012 patients were screened for inclusion. 462 were treated with raltegravir and 237 with placebo. At week 156, 51% in the raltegravir group versus 22% in the placebo group (non-completer classed as failure) had viral loads of less than 50 copies per mL, and 54% versus 23% had viral loads of less than 400 copies per mL. Mean CD4 cell count increase (analysed by an observed failure approach) was 164 cells per μL versus 63 cells per μL. After week 156, 251 patients (54%) from the raltegravir group and 47 (20%) from the placebo group entered the open-label raltergravir phase; 221 (47%) versus 44 (19%) completed the entire study. At week 240, viral load was less than 50 copies per mL in 193 (42%) of all patients initially assigned to raltegravir and less than 400 copies per mL in 210 (45%); mean CD4 cell count increased by 183 cells per μL. Virological failure occurred in 166 raltegravir recipients (36%) during the double-blind phase and in 17 of all patients (6%) during the open-label phase. The most common drug-related adverse events at 5 years in both groups were nausea, headache, and diarrhoea, and occurred in similar proportions in each group. Laboratory test results were similar in both treatment groups and showed little change after year 2. Interpretation: Raltegravir has a favourable long-term efficacy and safety profile in integrase-inhibitor-naive patients with triple-class resistant HIV in whom antiretroviral therapy is failing. Raltegravir is an alternative for treatment-experienced patients, particularly those with few treatment options. Funding: Merck Sharp & Dohme. © 2013 Elsevier Ltd.

Background - Acute heart failure is a common reason for admission, and outcome is often poor. Improved prognostic risk stratification may assist in the design of future trials and in patient management. Using data from a large randomized trial, we explored the prognostic value of clinical variables, measured at hospital admission for acute heart failure, to determine whether a few selected variables were inferior to an extended data set. Methods and Results - The prognostic model included 37 clinical characteristics collected at baseline in PROTECT, a study comparing rolofylline and placebo in 2033 patients admitted with acute heart failure. Prespecified outcomes at 30 days were death or rehospitalization for any reason; death or rehospitalization for cardiovascular or renal reasons; and, at both 30 and 180 days, all-cause mortality. No variable had a c-index >0.70, and few had values >0.60; c-indices were lower for composite outcomes than for mortality. Blood urea was generally the strongest single predictor. Eighteen variables contributed independent prognostic information, but a reduced model using only 8 items (age, previous heart failure hospitalization, peripheral edema, systolic blood pressure, serum sodium, urea, creatinine, and albumin) performed similarly. For prediction of all-cause mortality at 180 days, the model c-index using all variables was 0.72 and for the simplified model, also 0.72. Conclusions - A few simple clinical variables measured on admission in patients with acute heart failure predict a variety of adverse outcomes with accuracy similar to more complex models. However, predictive models were of only moderate accuracy, especially for outcomes that included nonfatal events. Better methods of risk stratification are required. © 2013 American Heart Association, Inc.

Siris E.S.,Columbia University | Modi A.,Merck And Co. | Tang J.,Asclepius JT LLC | Sen S.,Merck And Co.
Current Medical Research and Opinion | Year: 2014

Background: Multiple therapies are approved for the treatment of osteoporosis (OP), but many patients with osteoporosis may not initiate treatment upon osteoporosis diagnosis. Objective: To characterize initiation of pharmacologic OP treatment among women within 1 year of OP diagnosis in a US managed care population. Research design and methods: The retrospective cohort study included women aged 55 years with a claims-documented diagnosis of OP who were naïve to OP medications prior to OP diagnosis (index date) during 2001-2010. Continuous enrollment for 12 months before (baseline) and after (follow-up) the index date was required. Patients who received OP medications but did not have an OP diagnosis were excluded. Differences in baseline characteristics between the treated and untreated cohorts were compared using Wilcoxon rank-sum (continuous variables) and chi-square tests (categorical variables). Main outcomes measures: During the follow-up period, the percentages of patients treated with bisphosphonates (alendronate, ibandronate, risedronate, zoledronic acid) and non-bisphosphonates (calcitonin, raloxifene, teriparatide) were determined. Results: A total of 65,344 patients, mean age 65.7 years, met study inclusion exclusion criteria. During the follow-up period, 42,033 patients (64.3%) received no OP medication and 23,311 patients (35.7%) received OP treatment. A total of 20,200 patients (30.9% of total study population) received bisphosphonates and 3111 (4.8% of total) patients received non-bisphosphonates as their index medication. At baseline, untreated patients were slightly older and had higher rates of hypertension, chronic inflammatory joint disease, diabetes mellitus, and gastrointestinal events (p 0.01) compared with treated patients. Conclusions: Among women aged 55 years in a US managed-care population, 64.3% received no pharmacologic treatment within 1 year after being diagnosed with OP. The authors were not able to determine if untreated patients did not receive or did not fill a prescription. Further research is needed to understand the barriers to OP treatment and reasons for non-treatment. © 2014 Informa UK Ltd.

Clark K.,University of Dundee | Peggie M.,University of Dundee | Plater L.,University of Dundee | Sorcek R.J.,Boehringer Ingelheim Pharmaceuticals | And 6 more authors.
Biochemical Journal | Year: 2011

Members of the IKK {IκB [inhibitor of NF-κB (nuclear factor κB)] kinase} family play a central role in innate immunity by inducing NF-κB- and IRF [IFN (interferon) regulatory factor]-dependent gene transcription programmes required for the production of pro-inflammatory cytokines and IFNs. However, the molecular mechanisms that activate these protein kinases and their complement of physiological substrates remain poorly defined. Using MRT67307, a novel inhibitor of IKKε/TBK1 (TANK {TRAF [TNF (tumour-necrosis-factor)-receptor-associated factor]-associated NF-κB activator}-binding kinase 1) and BI605906, a novel inhibitor of IKKβ, we demonstrate that two different signalling pathways participate in the activation of the IKK-related protein kinases by ligands that activate the IL-1 (interleukin-1), TLR (Toll-like receptor) 3 and TLR4 receptors. One signalling pathway is mediated by the canonical IKKs,which directly phosphorylate and activate IKKε and TBK1, whereas the second pathway appears to culminate in the autocatalytic activation of the IKK-related kinases. In contrast, the TNFα-induced activation of the IKK-related kinases is mediated solely by the canonical IKKs. In turn, the IKK-related kinases phosphorylate the catalytic subunits of the canonical IKKs and their regulatory subunit NEMO (NF-κB essential modulator), which is associated with reduced IKKα/β activity and NF-κB-dependent gene transcription. We also show that the canonical IKKs and the IKK-related kinases not only have unique physiological substrates, such as IκBα, p105, RelA (IKKα and IKKβ) and IRF3 (IKKε and TBK1), but also have several substrates in common, including the catalytic and regulatory (NEMO and TANK) subunits of the IKKs themselves. Taken together, our studies reveal that the canonical IKKs and the IKK-related kinases regulate each other by an intricate network involving phosphorylation of their catalytic and regulatory (NEMO and TANK) subunits to balance their activities during innate immunity. © The Authors Journal compilation © 2011 Biochemical Society.

Sulkowski M.,Johns Hopkins University | Pol S.,University of Paris Descartes | Mallolas J.,University of Barcelona | Fainboim H.,Muniz Hospital | And 10 more authors.
The Lancet Infectious Diseases | Year: 2013

Background: Rates of sustained virological response (SVR) to peginterferon-ribavirin are low in patients with hepatitis C virus (HCV) genotype 1 and HIV. We aimed to assess efficacy and safety of triple therapy with boceprevir plus pegylated interferon alfa-2b (peginterferon) and ribavirin, which increases rates of SVR in patients with HCV alone. Methods: In our double-blind, randomised controlled phase 2 trial, we enrolled adults (18-65 years) with untreated HCV genotype 1 infection and controlled HIV (HIV RNA <50 copies per mL) at 30 academic and non-academic study sites. We randomly allocated patients (1:2) according to a computer generated sequence, stratified by Metavir score and baseline HCV RNA level, to receive peginterferon 1·5 μg/kg per week with weight-based ribavirin (600-1400 mg per day) for 4 weeks, followed by peginterferon-ribavirin plus either placebo (control group) or 800 mg boceprevir three times per day (boceprevir group) for 44 weeks. Non-nucleoside reverse-transcriptase inhibitors, zidovudine, and didanosine were not permitted. The primary efficacy endpoint was SVR (defined as undetectable plasma HCV RNA) at follow-up week 24 after end of treatment. We assessed efficacy and safety in all patients who received at least one dose of study drug. This study is registered with ClinicalTrials.gov, number NCT00959699. Findings: From Jan 15, 2010, to Dec 29, 2010, we enrolled 99 patients, 98 of whom received at least one treatment dose. 40 (63%) of 64 patients in the boceprevir group had an SVR at follow-up week 24, compared with ten (29%) of 34 control patients (difference 33·1%, 95% CI 13·7-52·5; p=0·0008). Adverse events were more common in patients who received boceprevir than in control patients: 26 (41%) versus nine (26%) had anaemia, 23 (36%) versus seven (21%) pyrexia, 22 (34%) versus six (18%) decreased appetite, 18 (28%) versus five (15%) dysgeusia, 18 (28%) versus five (15%) vomiting, and 12 (19%) versus two (6%) neutropenia. Three patients who received boceprevir plus peginterferon-ribavirin and four controls had HIV virological breakthrough. Interpretation: Boceprevir in combination with peginterferon-ribavirin could be an important therapeutic option for patients with HCV and HIV. Funding: Merck. © 2013 Elsevier Ltd.

Dan Y.Y.,National University of Singapore | Ferrante S.A.,Merck And Co. | Elbasha E.H.,Merck And Co. | Hsu T.-Y.,MSD Pharma Singapore Pte Ltd
Antiviral Therapy | Year: 2015

Background: Patients infected with chronic HCV genotype 1 experience liver complications as the disease progresses. This study aims to project the long-term reduction of liver complications and cost-effectiveness of treatment strategies, including co-administrating boceprevir (BOC) with pegylated interferon-α2b (PEG-IFN) and ribavirin compared with standard of care (SOC) of PEG-IFN and ribavirin only. Methods: A Markov model was created to estimate the expected costs and quality-adjusted life-years (QALYs) associated with treatment strategies outlined in the BOC package insert in Singapore. Patient characteristics were from pivotal trials, the transition probabilities and QALYs were estimated from publications, and the pharmaceutical and health status costs were obtained from a public hospital in Singapore. The threshold of cost-effectiveness was chosen as 65,000 SGD for this study. Results: For treatment-naive patients, BOC is highly cost-effective compared with SOC (179 SGD/QALY) and cost-saving for patients who have failed prior treatment, due to higher QALYs from better sustained virological response (SVR) and lower costs from avoidance of complications. Sub-group analyses show that BOC is cost-effective for non-cirrhotic treatment-experienced patients and null responders. It out-performs SOC for treatment-naive non-cirrhotic and cirrhotic patients who have failed prior treatment. Even after adjusting for higher prevalence of favourable IL28B genotype in Asians, BOC is cost-effective compared with SOC. Only untreated cirrhotic patients showed inconclusive cost-effectiveness for BOC. Conclusions: Compared with SOC, BOC prevents more HCV liver complications from HCV genotype 1, particularly in patients who have failed previous SOC. Improved SVR and shortened duration of treatment result in BOC being potentially cost-saving or cost-effective in an Asian population. ©2015 International Medical Press.

Background There is a high medical need for an interferon-free, all-oral, short-duration therapy for hepatitis C virus (HCV) that is highly effective across diverse patient populations, including patients with cirrhosis or previous null response to pegylated interferon (peginterferon) plus ribavirin (PR-null responders). We aimed to assess the efficacy, safety, and effective treatment duration of grazoprevir (an HCV NS3/4A protease inhibitor) combined with elbasvir (an HCV NS5A inhibitor) with or without ribavirin in patients with HCV genotype 1 infection with baseline characteristics of poor response. Methods The C-WORTHY trial is a randomised, open-label phase 2 trial of grazoprevir plus elbasvir with or without ribavirin; here we report findings for two cohorts of previously untreated patients with cirrhosis (cohort 1) and those with previous PR-null response with or without cirrhosis (cohort 2) enrolled in part B of the study. Eligible patients were adults aged 18 years or older with chronic HCV genotype 1 infection and HCV RNA concentrations of 10 000 IU/mL or higher in peripheral blood. We randomly assigned patients to receive grazoprevir (100 mg daily) and elbasvir (50 mg daily) with or without ribavirin for 12 or 18 weeks. Randomisation was done centrally with an interactive voice response system; patients and study investigators were masked to treatment duration up to week 12 but not to treatment allocation. The primary endpoint was the proportion of patients achieving HCV RNA less than 25 IU/mL at 12 weeks after end of treatment (SVR12), assessed by COBAS TaqMan version 2.0. This study is registered with ClinicalTrials.gov, number NCT01717326. Findings We describe findings for 253 patients enrolled in cohort 1 (n=123) or cohort 2 (n=130). In cohort 1, we randomly assigned 60 patients to the 12-week regimen (31 with ribavirin and 29 with no ribavirin) and 63 to the 18-week regimen (32 with ribavirin and 31 with no ribavirin); in cohort 2, we randomly assigned 65 patients to the 12-week regimen (32 with ribavirin and 33 with no ribavirin) and 65 to the 18-week regimen (33 with ribavirin and 32 with no ribavirin. High SVR12 rates were achieved irrespective of the use of ribavirin or extension of the treatment duration from 12 to 18 weeks; SVR12 rates ranged from 90% (95% CI 74-98; 28/31; cohort 1, 12 weeks, ribavirin-containing) to 100% (95% CI 89-100; 33/33; cohort 2, 18 weeks, ribavirin-containing). Among patients treated for 12 weeks with grazoprevir plus elbasvir without ribavirin, 97% (95% CI 82-100, 28/29) of patients in cohort 1 and 91% (76-98, 30/33) of patients in cohort 2 achieved SVR12. Adverse events reported in more than 10% of patients were fatigue (66 patients, 26% [95% CI 21-32]), headache (58 patients, 23% [95% CI 18-29]), and asthenia (35 patients, 14% [95% CI 10-19]). Interpretation Treatment with grazoprevir plus elbasvir, both with and without ribavirin and for both 12 and 18 weeks' treatment duration, showed high rates of efficacy in previously untreated patients with cirrhosis and previous PR-null responders with and without cirrhosis. These results support the phase 3 development of grazoprevir plus elbasvir. Funding Merck & Co, Inc. © 2015 Elsevier Ltd.

Wu G.,Peking University | Yin W.,Peking University | Shen H.C.,Merck And Co. | Huang Y.,Peking University
Green Chemistry | Year: 2012

To access useful heterocycles in medicinal chemistry such as pyridazinones, dihydropyrimidinones, and dihydropyrimidinthiones, a "green" mild and highly efficient one-pot triple cascade was developed involving a Claisen-decarboxylation, electrophilic reaction, and subsequent heterocyclization. In addition, indazoles and benzofurans could also be constructed via a double cascade. To develop the cascade process, a direct Claisen-decarboxylation reaction was firstly optimized. This reaction can then couple with electrophilic reactions including alkylation, Michael addition or aldol reaction to enable the preparation of various aryl ketones in a one-pot fashion. © The Royal Society of Chemistry 2012.

Massie B.M.,University of California at San Francisco | Massie B.M.,San Francisco Medical Center | O'Connor C.M.,Duke University | Metra M.,University of Brescia | And 13 more authors.
New England Journal of Medicine | Year: 2010

Background: Worsening renal function, which is associated with adverse outcomes, often develops in patients with acute heart failure. Experimental and clinical studies suggest that counterregulatory responses mediated by adenosine may be involved. We tested the hypothesis that the use of rolofylline, an adenosine A1-receptor antagonist, would improve dyspnea, reduce the risk of worsening renal function, and lead to a more favorable clinical course in patients with acute heart failure. Methods: We conducted a multicenter, double-blind, placebo-controlled trial involving patients hospitalized for acute heart failure with impaired renal function. Within 24 hours after presentation, 2033 patients were randomly assigned, in a 2:1 ratio, to receive daily intravenous rolofylline (30 mg) or placebo for up to 3 days. The primary end point was treatment success, treatment failure, or no change in the patient's clinical condition; this end point was defined according to survival, heart-failure status, and changes in renal function. Secondary end points were the post-treatment development of persistent renal impairment and the 60-day rate of death or readmission for cardiovascular or renal causes. Results: Rolofylline, as compared with placebo, did not provide a benefit with respect to the primary end point (odds ratio, 0.92; 95% confidence interval, 0.78 to 1.09; P = 0.35). Persistent renal impairment developed in 15.0% of patients in the rolofylline group and in 13.7% of patients in the placebo group (P = 0.44). By 60 days, death or readmission for cardiovascular or renal causes had occurred in similar proportions of patients assigned to rolofylline and placebo (30.7% and 31.9%, respectively; P = 0.86). Adverse-event rates were similar overall; however, only patients in the rolofylline group had seizures, a known potential adverse effect of A1-receptor antagonists. Conclusions: Rolofylline did not have a favorable effect with respect to the primary clinical composite end point, nor did it improve renal function or 60-day outcomes. It does not show promise in the treatment of acute heart failure with renal dysfunction. Copyright © 2010 Massachusetts Medical Society.

Khan K.H.,GI and Lymphoma Unit | Yap T.A.,The Royal Marsden NHS Foundation Trust | Yan L.,Merck And Co. | Yan L.,Peking University | Cunningham D.,GI and Lymphoma Unit
Chinese Journal of Cancer | Year: 2013

The phosphoinositide 3-kinase-AKT-mammalian target of rapamycin (PI3K-AKT-mTOR) pathway is a frequently hyperactivated pathway in cancer and is important for tumor cell growth and survival. The development of targeted therapies against mTOR, a vital substrate along this pathway, led to the approval of allosteric inhibitors, including everolimus and temsirolimus, for the treatment of breast, renal, and pancreatic cancers. However, the suboptimal duration of response in unselected patients remains an unresolved issue. Numerous novel therapies against critical nodes of this pathway are therefore being actively investigated in the clinic in multiple tumour types. In this review, we focus on the progress of these agents in clinical development along with their biological rationale, the need of predictive biomarkers and various combination strategies, which will be useful in counteracting the mechanisms of resistance to this class of drugs.

Herbst R.S.,Yale University | Baas P.,Netherlands Cancer Institute | Kim D.-W.,Seoul National University | Felip E.,University of Barcelona | And 15 more authors.
The Lancet | Year: 2016

Background Despite recent advances in the treatment of advanced non-small-cell lung cancer, there remains a need for effective treatments for progressive disease. We assessed the efficacy of pembrolizumab for patients with previously treated, PD-L1-positive, advanced non-small-cell lung cancer. Methods We did this randomised, open-label, phase 2/3 study at 202 academic medical centres in 24 countries. Patients with previously treated non-small-cell lung cancer with PD-L1 expression on at least 1% of tumour cells were randomly assigned (1:1:1) in blocks of six per stratum with an interactive voice-response system to receive pembrolizumab 2 mg/kg, pembrolizumab 10 mg/kg, or docetaxel 75 mg/m2 every 3 weeks. The primary endpoints were overall survival and progression-free survival both in the total population and in patients with PD-L1 expression on at least 50% of tumour cells. We used a threshold for significance of p<0·00825 (one-sided) for the analysis of overall survival and a threshold of p<0·001 for progression-free survival. This trial is registered at ClinicalTrials.gov, number NCT01905657. Findings Between Aug 28, 2013, and Feb 27, 2015, we enrolled 1034 patients: 345 allocated to pembrolizumab 2 mg/kg, 346 allocated to pembrolizumab 10 mg/kg, and 343 allocated to docetaxel. By Sept 30, 2015, 521 patients had died. In the total population, median overall survival was 10·4 months with pembrolizumab 2 mg/kg, 12·7 months with pembrolizumab 10 mg/kg, and 8·5 months with docetaxel. Overall survival was significantly longer for pembrolizumab 2 mg/kg versus docetaxel (hazard ratio [HR] 0·71, 95% CI 0·58-0·88; p=0·0008) and for pembrolizumab 10 mg/kg versus docetaxel (0·61, 0·49-0·75; p<0·0001). Median progression-free survival was 3·9 months with pembrolizumab 2 mg/kg, 4·0 months with pembrolizumab 10 mg/kg, and 4·0 months with docetaxel, with no significant difference for pembrolizumab 2 mg/kg versus docetaxel (0·88, 0·74-1·05; p=0·07) or for pembrolizumab 10 mg/kg versus docetaxel (HR 0·79, 95% CI 0·66-0·94; p=0·004). Among patients with at least 50% of tumour cells expressing PD-L1, overall survival was significantly longer with pembrolizumab 2 mg/kg than with docetaxel (median 14·9 months vs 8·2 months; HR 0·54, 95% CI 0·38-0·77; p=0·0002) and with pembrolizumab 10 mg/kg than with docetaxel (17·3 months vs 8·2 months; 0·50, 0·36-0·70; p<0·0001). Likewise, for this patient population, progression-free survival was significantly longer with pembrolizumab 2 mg/kg than with docetaxel (median 5·0 months vs 4·1 months; HR 0·59, 95% CI 0·44-0·78; p=0·0001) and with pembrolizumab 10 mg/kg than with docetaxel (5·2 months vs 4·1 months; 0·59, 0·45-0·78; p<0·0001). Grade 3-5 treatment-related adverse events were less common with pembrolizumab than with docetaxel (43 [13%] of 339 patients given 2 mg/kg, 55 [16%] of 343 given 10 mg/kg, and 109 [35%] of 309 given docetaxel). Interpretation Pembrolizumab prolongs overall survival and has a favourable benefit-to-risk profile in patients with previously treated, PD-L1-positive, advanced non-small-cell lung cancer. These data establish pembrolizumab as a new treatment option for this population and validate the use of PD-L1 selection. © 2016 Elsevier Ltd.

Arruda M.A.,Glia Institute | Guidetti V.,University of Rome La Sapienza | Galli F.,University of Rome La Sapienza | Bigal M.E.,Merck And Co. | Bigal M.E.,Yeshiva University
Cephalalgia | Year: 2010

The aim of this study was to estimate the prevalence of primary headaches in pre-adolescent children, as well as headache frequency and days of treatment in this population. Sample consisted of 1994 children (aged 5-12 years). Parents were interviewed by a paediatric headache specialist using a questionnaire that allowed the classification of headaches using the criteria of the Second Edition of the International Classification of Headache Disorders. The most severe headache type was classified (mutually-exclusive diagnoses). Prevalence and prevalence ratios were calculated overall, as well as by age, gender and race. The overall prevalence of migraine was 3.76%, non-significantly higher in boys (3.9%) than in girls (3.6%). Prevalence of probable migraine was significantly higher than the prevalence of migraine for all ages (overall prevalence of 17.1%). Chronic migraine (CM) happened in 0.8% (girls, 1.15%; boys, 0.5%). Infrequent episodic tension-type headache (ETTH) happened in 2.3% of the sample while prevalence of frequent ETTH was 1.6%. Probable TTH happened in 13.5%. Most children with migraine had consulted a medical doctor because of their headaches, and the proportion was higher among children with CM (93.7%). Prevalence of primary headaches is high in young children. Probable diagnoses are more common than full diagnoses. Consultation rates are elevated. © International Headache Society 2010.

Johnson E.L.,Johns Hopkins University | Chang Y.-T.,Johnson and Johnson | Davit B.,Merck And Co. | Gidal B.E.,University of Wisconsin - Madison | Krauss G.L.,Johns Hopkins University
Neurology | Year: 2016

Objectives: The purpose of this study was to determine how closely generic modified-release antiepileptic drugs (MR-AEDs) resemble reference (brand) formulations by comparing peak concentrations (Cmax), total absorption (area under the curve [AUC]), time to Cmax (Tmax), intersubject variability, and food effects between generic and reference products. Methods: We tabulated Cmax and AUC data from the bioequivalence (BE) studies used to support the approvals of generic Food and Drug Administration-approved MR-AEDs. We compared differences in 90% confidence intervals of the generic/reference AUC and Cmax geometric mean ratios, and intersubject variability, Tmax and delivery profiles and food effects. Results: Forty-two MR-AED formulations were studied in 3,175 healthy participants without epilepsy in 97 BE studies. BE ratios for AUC and Cmax were similar between most generic and reference products: AUC ratios varied by >15% in 11.4% of BE studies; Cmax varied by >15% in 25.8% of studies. Tmax was more variable, with >30% difference in 13 studies (usually delayed in the fed compared to fasting BE studies). Generic and reference MR products had similar intersubject variability. Immediate-release AEDs showed less intersubject variability in AUC than did MR-AEDs. Conclusions: Most generic and reference MR-AEDs have similar AUC and Cmax values. Ratios for some products, however, are near acceptance limits and Tmax values may vary. Food effects are common with MR-AED products. High variability in pharmacokinetic values for once-a-day MR-AEDs suggests their major advantage compared to immediate-release AED formulations may be the convenience of less frequent dosing to improve adherence. © 2016 American Academy of Neurology.

Patel K.,U.S. National Institute on Aging | Faulkner K.,University of Pittsburgh | Inzitari M.,Autonomous University of Barcelona | Chandler J.,Merck And Co. | And 9 more authors.
JAMA - Journal of the American Medical Association | Year: 2011

Context: Survival estimates help individualize goals of care for geriatric patients, but life tables fail to account for the great variability in survival. Physical performance measures, such as gait speed, might help account for variability, allowing clinicians to make more individualized estimates. Objective: To evaluate the relationship between gait speed and survival. Design, Setting, and Participants: Pooled analysis of 9 cohort studies (collected between 1986 and 2000), using individual data from 34 485 community-dwelling older adults aged 65 years or older with baseline gait speed data, followed up for 6 to 21 years. Participants were a mean (SD) age of 73.5 (5.9) years; 59.6%, women; and 79.8%, white; and had a mean (SD) gait speed of 0.92 (0.27) m/s. Main Outcome Measures: Survival rates and life expectancy. Results: There were 17 528 deaths; the overall 5-year survival rate was 84.8% (confidence interval [CI],79.6%-88.8%)and 10-year survival ratewas59.7%(95%CI,46.5%-70.6%). Gait speed was associated with survival in all studies (pooled hazard ratio per 0.1 m/s, 0.88; 95% CI, 0.87-0.90; P<.001). Survival increased across the full range of gait speeds, with significant increments per 0.1 m/s. At age 75, predicted 10-year survival across the range of gait speeds ranged from 19% to 87% in menand from35% to 91% in women. Predicted survival based on age, sex, and gait speed was as accurate as predicted based on age, sex, use of mobility aids, and self-reported function or as age, sex, chronic conditions, smoking history, blood pressure, body mass index, and hospitalization. Conclusion: In this pooled analysis of individual data from 9 selected cohorts, gait speed was associated with survival in older adults. ©2011 American Medical Association. All rights reserved.

Sistare F.D.,Merck And Co. | DeGeorge J.J.,Merck And Co.
Toxicologic Pathology | Year: 2015

High-dose selection for 6-month carcinogenicity studies of pharmaceutical candidates in Tg.rasH2-transgenic mice currently primarily relies on (1) estimation of a maximum tolerated dose (MTD) from the results of a 1-month range-finding study, (2) determination of the maximum dose administrable to the animals (maximum feasible dose [MFD]), (3) demonstration of a plateau in systemic exposure, and (4) use of a limit dose of 1,500 mg/kg/day for products with human daily doses not exceeding 500 mg. Eleven 6-month Tg.rasH2 carcinogenicity studies and their corresponding 1-month range-finding studies conducted at Merck were reviewed. High doses were set by estimation of the MTD in 6, by plateau of exposure in 3, and by MFD in 2 cases. For 4 of 6 studies where MTD was used for high-dose selection, the 1-month study accurately predicted the 6-month study tolerability whereas in the remaining 2 studies the high doses showed poorer tolerability than expected. The use of 3 or more drug-treated dose levels proved useful to ensure that a study would successfully and unambiguously demonstrate that a drug candidate was adequately evaluated for carcinogenicity at a minimally toxic high dose level, especially when the high dose may be found to exceed the MTD. Copyright © 2015 by The Author(s).

Ho G.Y.F.,Yeshiva University | Wang T.,Yeshiva University | Gunter M.J.,Imperial College London | Strickler H.D.,Yeshiva University | And 8 more authors.
Cancer Research | Year: 2012

Mechanistic associations between obesity and colorectal cancer remain unclear. In this study, we investigated whether adipokines are risk factors for colorectal cancer and whether they may mediate its association with obesity. In a case-cohort study nested within the Women's Health Initiative cohort of postmenopausal women, baseline plasma samples from 457 colorectal cancer cases and 841 subcohort subjects were assayed for seven adipokines - adiponectin, leptin, plasminogen activator inhibitor-1 (PAI-1), resistin, hepatocyte growth factor, interleukin-6 (IL-6), and TNF-α. Serum insulin and estradiol values measured previously were also available for data analysis. After adjusting for age, race, smoking, colonoscopy history, and estrogen level, a low level of anti-inflammatory adiponectin and high levels of proinflammatory leptin, PAI-1, and IL-6 were associated with increased colorectal cancer risk, though only leptin remained significant after further adjustment for insulin [HRs comparing extreme quartiles (HRQ4-Q1), 1.84; 95% CI, 1.17-2.90]. Mediation analyses showed that leptin and insulin partially explained the association between waist circumference and colorectal cancer and attenuated it by 25% and 37%, respectively, with insulin being a significant mediator (P = 0.041). Our findings support the conclusion that adipokines involved in inflammation are associated with colorectal cancer risk, but that their effects may be mediated mostly by insulin, with leptin exerting an independent effect. Hyperinsulinemia and hyperleptinemia may therefore partially explain the adiposity association with colorectal cancer in postmenopausal women. ©2012 AACR.

Kemper K.,Center for Experimental and Molecular Medicine | Sprick M.R.,Center for Experimental and Molecular Medicine | De Bree M.,Center for Experimental and Molecular Medicine | Scopelliti A.,Cellular and Molecular Pathophysiology Laboratory | And 5 more authors.
Cancer Research | Year: 2010

Colon cancer stem cells (CSC) can be identified with AC133, an antibody that detects an epitope on CD133. However, recent evidence suggests that expression of CD133 is not restricted to CSCs, but is also expressed on differentiated tumor cells. Intriguingly, we observed that detection of the AC133 epitope on the cell surface decreased upon differentiation of CSC in a manner that correlated with loss of clonogenicity. However, this event did not coincide with a change in CD133 promoter activity, mRNA, splice variant, protein expression, or even cell surface expression of CD133. In contrast, we noted that with CSC differentiation, a change occured in CD133 glycosylation. Thus, AC133 may detect a glycosylated epitope, or differential glycosylation may cause CD133 to be retained inside the cell. We found that AC133 could effectively detect CD133 glycosylation mutants or bacterially expressed unglycosylated CD133. Moreover, cell surface biotinylation experiments revealed that differentially glycosylated CD133 could be detected on the membrane of differentiated tumor cells. Taken together, our results argue that CD133 is a cell surface molecule that is expressed on both CSC and differentiated tumor cells, but is probably differentially folded as a result of differential glycosylation to mask specific epitopes. In summary, we conclude that AC133 can be used to detect cancer stem cells, but that results from the use of this antibody should be interpreted with caution. ©2010 AACR.

Rockstroh J.K.,University of Bonn | Nelson M.,Chelsea and Westminster Hospital | Katlama C.,French Institute of Health and Medical Research | Katlama C.,APHP Pitie Salpetriere Hospital | And 16 more authors.
The Lancet HIV | Year: 2015

Background Hepatitis C virus (HCV) infection is a leading cause of morbidity and mortality in patients with HIV-1. The C-EDGE CO-INFECTION study assessed the effi cacy, safety, and tolerability of grazoprevir (MK-5172) plus elbasvir (MK-8742) in patients with HCV and HIV co-infection. Methods In this uncontrolled, non-randomised, phase 3, open-label, single-arm study, treatment-naive patients with chronic HCV genotype 1, 4, or 6 infection and HIV co-infection, with or without cirrhosis, were enrolled from 37 centres in nine countries across Europe, the USA, and Australia. Patients were either naive to treatment with any antiretroviral therapy (ART) or stable on ART for at least 8 weeks. All patients received grazoprevir 100 mg plus elbasvir 50 mg in a fi xed-dose combination tablet once daily for 12 weeks. The primary endpoint was sustained virological response (HCV RNA <15 IU/mL) 12 weeks after the end of therapy (SVR12). The primary population for effi cacy analyses was all patients who received at least one dose of study treatment. This study is registered with ClinicalTrials.gov, number NCT02105662. Findings Between June 11, 2014, and Aug 29, 2014, 218 patients were enrolled and received grazoprevir plus elbasvir for 12 weeks, all of whom completed follow-up at week 12. SVR12 was achieved by 210 (96%) of 218 patients (95% CI 92.9-98.4). One patient did not achieve SVR12 because of a non-virological reason, and seven patients without cirrhosis relapsed (two subsequently confi rmed as reinfections). All 35 patients with cirrhosis achieved SVR12. The most common adverse events were fatigue (29; 13%), headache (27; 12%), and nausea (20; 9%). No patient discontinued treatment because of an adverse event. Two patients receiving ART had transient HIV viraemia. Interpretation This HCV treatment regimen seems to be eff ective and well tolerated for patients co-infected with HIV with or without cirrhosis. These data are consistent with previous trials of this regimen in the monoinfected population. This regimen continues to be studied in phase 3 trials.

Bland C.S.,Baylor College of Medicine | Wang E.T.,Massachusetts Institute of Technology | David M.P.,University of Arkansas for Medical Sciences | Castle J.C.,Institute for Translational Oncology and Immunology | And 3 more authors.
Nucleic Acids Research | Year: 2010

Recent genome-wide analyses have elucidated the extent of alternative splicing (AS) in mammals, often focusing on comparisons of splice isoforms between differentiated tissues. However, regulated splicing changes are likely to be important in biological transitions such as cellular differentiation, or response to environmental stimuli. To assess the extent and significance of AS in myogenesis, we used splicing-sensitive microarray analysis of differentiating C2C12 myoblasts. We identified 95 AS events that undergo robust splicing transitions during C2C12 differentiation. More than half of the splicing transitions are conserved during differentiation of avian myoblasts, suggesting the products and timing of transitions are functionally significant. The majority of splicing transitions during C2C12 differentiation fall into four temporal patterns and were dependent on the myogenic program, suggesting that they are integral components of myogenic differentiation. Computational analyses revealed enrichment of many sequence motifs within the upstream and downstream intronic regions near the alternatively spliced regions corresponding to binding sites of splicing regulators. Western analyses demonstrated that several splicing regulators undergo dynamic changes in nuclear abundance during differentiation. These findings show that within a developmental context, AS is a highly regulated and conserved process, suggesting a major role for AS regulation in myogenic differentiation. © 2010 The Author(s).

Peacock H.,University of California at Davis | Fucini R.V.,Merck And Co. | Jayalath P.,University of California at Davis | Ibarra-Soza J.M.,University of California at Davis | And 4 more authors.
Journal of the American Chemical Society | Year: 2011

Immune stimulation is a significant hurdle in the development of effective and safe RNA interference therapeutics. Here, we address this problem in the context of a mimic of microRNA-122 by employing novel nucleobase and known 2′-ribose modifications. The nucleobase modifications are analogues of adenosine and guanosine that contain cyclopentyl and propyl minor-groove projections. Via a site-by-site chemical modification analysis, we identify several immunostimulatory hot spots within the miRNA guide strand at which single base modifications significantly reduce immune stimulation. A duplex containing one base modification on each strand proved to be most effective in preventing immune stimulation. © 2011 American Chemical Society.

Atack J.R.,Merck And Co. | Hargreaves R.J.,Merck And Co. | Burns H.D.,Merck And Co. | Dawson G.R.,Merck And Co.
Journal of Pharmacology and Experimental Therapeutics | Year: 2010

Abuse-liability-related effects of subtype-selective GABAA modulators were explored relative to the prototypic benzodiazepine lorazepam. 7-Cyclobutyl-6-(2-methyl-2H-1,2,4-triazol-3-ylmethoxy)-3-phenyl-1,2, 4-triazolo[4,3-b]pyridazine (TPA123) has weak partial agonist efficacy at α1-, α2-, α3-, and α5-containing GABAA receptors, whereas 7-(1,1-dimethylethyl)-6-(2-ethyl-2H-1,2,4-triazol-3-ylmethoxy) -3-(2-fluorophenyl)-1,2,4-triazolo[4,3-b]pyridazine (TPA023) has weaker partial agonist efficacy at α2 and α3 and none at α1 and α5 subtypes. For both compounds, preclinical data suggested efficacy as nonsedating anxiolytics. Self-injection of TPA123 (0.0032- 0.1 mg/kg) and TPA023 (0.0032- 0.32 mg/ kg) was compared with lorazepam (0.01- 0.32 mg/kg) in baboons. TPA123 and lorazepam maintained self-injection higher than vehicle at two or more doses in each baboon; peak rate of self-injection of lorazepam was higher than TPA123. Self-injected lorazepam and TPA123 also increased rates of concurrently occurring food-maintained behavior. After the availability of self-administered TPA123 doses ended, an effect consistent with a mild benzodiazepine-like withdrawal syndrome occurred. In contrast with lorazepam and TPA123, TPA023 did not maintain self-administration. Positron emission tomography studies showed that TPA023 produced a dose-dependent inhibition in the binding of [ 11C]flumazenil to the benzodiazepine binding site in the baboon, which was essentially complete (i.e., 100% occupancy) at the highest TPA023 dose (0.32 mg/kg). In a physical dependence study, TPA023 (32 mg/kg/24 h) was delivered as a continuous intragastric drip. Neither flumazenil at 14 days nor stopping TPA023 after 30 to 31 days resulted in the marked withdrawal syndrome characteristic of benzodiazepines in baboons. In the context of other data, elimination of efficacy at the α1 subtype of the GABA/benzodiazepine receptor is not sufficient to eliminate abuse liability but may do so when coupled with reduced α2/3 subtype efficacy. Copyright © 2010 by The American Society for Pharmacology and Experimental Therapeutics.

Yin D.D.,Merck And Co. | Nocea G.,Outcomes Research | Qiu Y.,Merck And Co. | Mavros P.,Merck And Co.
Health and Quality of Life Outcomes | Year: 2010

Background: To evaluate the association between patient-reported hypoglycemic symptoms with ratings of their health-related quality of life state and patient-reported adverse events in patients with type 2 diabetes mellitus (T2DM).Methods: This observational, multicenter, cross sectional study was based on a sample of patients with T2DM from seven European countries who added sulfonylurea or thiazolidinedione to metformin monotherapy between January 2001 and January 2006. Included patients were required to have at least one hemoglobin A 1c(HbA 1c) measurement in the 12 months before enrollment and to not be receiving insulin. Demographic and clinical data from medical records were collected using case report forms. Questionnaires measured patient-reported hypoglycemic symptoms, health-related quality of life (EuroQol visual analogue scale, EQ-5D VAS), and treatment-related adverse events.Results: A total of 1,709 patients were included in the study. Mean patient age was 63 years, 45% were female, mean HbA 1cwas 7.06%, and 28% were at HbA 1cgoal (HbA 1c< 6.5%). Hypoglycemic symptoms during the 12 months before enrollment were reported by 38% of patients; among whom 68% reported their most severe symptoms were mild, 27% moderate, and 5% severe. Adjusted linear regression analyses revealed that patients reporting hypoglycemic symptoms had significantly lower EQ-5D VAS scores indicating worse patient-reported quality of life (mean difference -4.33, p < 0.0001). Relative to those not reporting symptoms, the adjusted decrement to quality of life increased with greater hypoglycemic symptom severity (mild: -2.68, p = 0.0039; moderate: -6.42, p < 0.0001; severe: -16.09, p < 0.0001). Patients with hypoglycemia reported significantly higher rates of shakiness, sweating, excessive fatigue, drowsiness, inability to concentrate, dizziness, hunger, asthenia, and headache (p < 0.0001 for each comparison).Conclusions: Hypoglycemic symptoms and symptom severity have an adverse effect on patients' rating of their health related quality of life state. Hypoglycemic symptoms are correlated with treatment-related adverse effects. Minimizing the risk and severity of hypoglycemia may improve patients' quality of life and clinical outcomes. Results are subject to limitations associated with observational studies including the potential biases due to unobserved patient heterogeneity and the use of a convenience sample of patients. © 2010 Alvarez-Guisasola et al; licensee BioMed Central Ltd.

Sicouri S.,Masonic Medical Research Laboratory | Pourrier M.,Cardiome Pharma | Gibson J.K.,AAKVSL Pharma Consulting LLC | Lynch J.J.,Merck And Co. | Antzelevitch C.,Masonic Medical Research Laboratory
Heart Rhythm | Year: 2012

Background: Vernakalant (VER) is a relatively atrial-selective antiarrhythmic drug capable of blocking potassium and sodium currents in a frequency- and voltage-dependent manner. Ranolazine (RAN) is a sodium-channel blocker shown to exert antiarrhythmic effects in pulmonary vein (PV) sleeves. dl-Sotalol (SOT) is a β-blocker commonly used in the rhythm-control treatment of atrial fibrillation. This study evaluated the electrophysiological and antiarrhythmic effects of VER, RAN, and SOT in canine PV sleeve preparations in a blinded fashion. Methods: Transmembrane action potentials were recorded from canine superfused PV sleeve preparations exposed to VER (n = 6), RAN (n = 6), and SOT (n = 6). Delayed afterdepolarizations were induced in the presence of isoproterenol and high-calcium concentrations by periods of rapid pacing. Results: In PV sleeves, VER, RAN, and SOT (330 μM) produced small (1015 ms) increases in action potential duration. The effective refractory period, diastolic threshold of excitation, and the shortest S1S1 cycle length permitting 1:1 activation were significantly increased by VER and RAN in a rate- and concentration-dependent manner. VER and RAN significantly reduced Vmax in a concentration- and rate-dependent manner. SOT did not significantly affect the effective refractory period, Vmax, diastolic threshold of excitation, or the shortest S1S1 cycle length permitting 1:1 activation. All 3 agents (330 μM) suppressed delayed afterdepolarizationmediated triggered activity induced by isoproterenol and high calcium. Conclusions: In canine PV sleeves, the effects of VER and RAN were similar and largely characterized by concentration- and rate-dependent depression of sodium-channelmediated parameters, which were largely unaffected by SOT. All 3 agents demonstrated an ability to effectively suppress delayed afterdepolarizationinduced triggers of atrial arrhythmia. © 2012 Heart Rhythm Society. All rights reserved.

Eduardo Bigal M.,Merck And Co.
Clinical Journal of Pain | Year: 2010

Objectives: To identify signs of temporomandibular disorders and cervical pain in individuals with episodic and chronic (transformed) migraine (CM), relative to controls without headaches. Methods: In this prospective, controlled, double-blind study, we examined 93 individuals divided in 3 groups: episodic migraine EM, (n=31), CM chronic migraine (n=34), and controls without migraine (n=28). We recorded signs of temporomandibular disorders, and of pain in the neck, after the protocol of Helkimo (1974). We calculated the odds ratio (OR) and confidence intervals (CI) of symptoms as a function of headache status. Data from all groups were paired and compared using the χ2 test. The level of significance was 5% in 2-tailed tests. Results: Relative to controls, participants with EM and CM were significantly more likely to have tenderness in the masticatory muscles [controls=28%, migraine=54%, (OR=3.0, 95% CI=1.1-8.9), CM=73% (OR=6.9, 95% CI=2.3-21.2)], and in the temporomandibular joint [controls=25%, migraine=61%, (OR=4.7, 95% CI=1.5-14.5), CM=61% (OR=4.8, 95% CI=1.6-14.5)]. They were numerically (but nonsignificantly) more likely to have limited lateral jaw movements (CM=34%; EM=26%; NP=18%), joint sounds (CM=44%; EM=29%; NP=28%), and tenderness in neck muscles (CM=64%; EM=51%; NP=35%). Conclusion: In a tertiary care population, individuals with EM and CM are more likely to have tenderness at the temporomandibular joint and on the masticatory muscles, relative to controls. Studies are needed to investigate whether treatment of 1 disorder will improve the other. © 2010 by Lippincott Williams & Wilkins.

Agency: European Commission | Branch: FP7 | Program: CP-IP | Phase: NMP-2007-2.2-1 | Award Amount: 27.17M | Year: 2009

The call 4.2.2-1 organic materials for electronics and photonics is based on the observation that the limited availability of high-performance multi-functional materials is a roadblock to further industrial progress. To address the wide scope of the call, we have identified specific materials bottlenecks to the fields of electronics and photonics. They constitute the focal points of our project. One-P main objective is: to invent, design, synthesize, characterize, process, and to supply the missing materials in the fields of organic electronics and photonics and to develop appropriate patterning methods for micro- and nano-structuring of these materials that can be up-scaled to roll-to-roll technologies. The work plan is composed of five technical workpackages, each one addressing current materials challenges: 1) charge transport and injection, 2) detection and sensing, 3) light emission, 4) functional self-assembled monolayers, 5) continuous processing and technology. Computer-aided design of materials and the use of advanced characterization tools are transversal activities that are integrated in technical workpackages. The sixth workpackage is devoted to dissemination, exploitation, and management of intellectual properties that are essential for the project success. To carry out this multi-disciplinary project, a cross-sectorial consortium has been formed at the European level. It is composed of strong academic and industrial teams with necessary and complementary expertises to cover all scientific, technological and exploitation aspects. The project will generate fundamental knowledge and help to develop unprecedented technologies. They will have a positive impact on competitiveness of European industries, environment, job creation, health, security, safety, and welfare of European citizens

Agency: European Commission | Branch: FP7 | Program: MC-ITN | Phase: PEOPLE-2007-1-1-ITN | Award Amount: 4.65M | Year: 2008

The mission of HIERARCHY is to train and educate young scientist in the rapidly developing field of nanosciences, in particular hierarchical self-assembly. The training programme educates early stage and experienced researches in many aspects of this highly interdisciplinary field, such as theory, materials chemistry and biochemistry, advanced characterisation techniques, physics and commercial device development. In addition, the training programme will address non-scientific issues, important for the career development of young scientists, e.g. communication and presentation skills, IPR and entrepreneurial skills, ethical issues, language enhancement and cultural awareness. The training takes place on a Network level and also locally at the host institutions. HIERARCHYs training programme will deliver versatile individuals with a broad scientific knowledge, ready to pursue a successful career in the European industry or academia. The interdisciplinary research training is centralised around the novel concept of hierarchical assembly in controllable matrices. This concept exploits liquid crystalline media as controllable matrices for programmed self-organisation, which goes far beyond the possibilities of currently employed techniques. A liquid crystal matrix in combination with a variety of simultaneously or sequentially applied external stimuli will yield a unique toolbox to build functional macroscopic structures with nanometer control. Leading European laboratories in soft condensed matter and solid state matter will work towards new paradigms in nanosciences. HIERARCHYs intention towards application of the designed structures, illustrated by the presence of three industrial partners in the consortium, is an important step towards commercialisation of nanosciences in Europe. With Europes desire to become the major player in the area of nanosciences, valorisation of developed technology is a key lesson for Europes new generation of nanoscientists.

Agency: GTR | Branch: EPSRC | Program: | Phase: Research Grant | Award Amount: 5.84M | Year: 2011

In the 1980s it began to be possible to produce potentially unlimited quantities of human proteins by placing the gene defining them in a simple organism such as yeast. From this grew a new kind of medicine capable of treating conditions such as severe arthritis, haemophilia, growth deficiency, and some cancers that previously had no satisfactory treatments. As well as having great clinical value the resulting technology has become the basis of a new and fastest growing part of the pharmaceutical industry, described as biopharmaceuticals. Because the molecules involved are proteins, they are orders of magnitude larger and more complex than conventional drugs such as aspirin and their processing is much more demanding. They are also so complex that they cannot in general be characterised with precision except in relation to the methods by which they are made. That means the capacity to precisely define such processes is critical to clinical safety and commercial success. Full scale trials of the processes are so costly they can only be conducted once clinical promise is established but, given the number of factors governing processing of even first generation products, there have often been hold-ups so extensive as to delay availability to patients. UCL has pioneered micro scale methods that are sufficiently good at predicting efficient conditions for large scale performance that far fewer and better focussed large scale trials suffice. That resolves part of the problem but an even greater challenge is now emerging. The early biopharmaceuticals were in general the easiest ones to produce. The final scales were also relatively modest. Now, the next generation of biopharmaceuticals are more complex materials and with rising demand the scales are far larger so that processes push the boundaries of the possible. The combined complexity of the product and the process with so many variables to consider means that the managers need better systematic means of supporting their decisions. Already the cost of developing a single biopharmaceutical can exceed 0.7 billion and take 10 years. With more advanced biopharmaceuticals these figures tend to rise and yet the worlds governments are facing a healthcare cost crisis with more older people. They therefore exert pressure on companies to reduce prices. Because the public wishes to have medicines that do not pose risks, regulations become ever more stringent so they are a major factor in defining the bioprocess. This also adds to the need for managers to have sector-specific decisional-support aids well grounded in the detailed engineering of the processes. Finally, it is now possible to apply molecular engineering to proteins and vaccines to enhance their therapeutic properties but this can also cause serious bioprocessing problems. The research vision developed with detailed input from UK industry experts will apply these methods as the foundation for another step change whereby much faster and lower cost information can be gathered and integrated with advanced decisional techniques to give managers a better foundation on which to base their policies. The academic team from leading UK universities provides the necessary continuum of skills needed to assess the ease of manufacture of novel drugs, the costs of processing and of delivery to patients. We will work with companies to test the outcomes to ensure they are well proven prior to use on new biopharmaceuticals. This will cut costs so that all the patients who might benefit can receive them and at the earliest possible date achieved within the severely restricted budgets now available to the NHS.

Radican L.,Merck And Co. | Kong S.X.,Novo Nordisk AS
Journal of Medical Economics | Year: 2013

Objective: Understanding of the effects of providers' cost on regional variation in healthcare spending is still very limited. The objective of this study is to assess cross-state and cross-region variations in inpatient cost of lower extremity amputation among diabetic patients (DLEA) in relation to patient, hospital, and state factors. Methods: Patient and hospital level data were obtained from the 2007 US Agency for Healthcare Research and Quality Healthcare Cost and Utilization Project (HCUP). State level data were obtained from the US Census Bureau and the Kaiser Family Foundation websites. Regression models were implemented to analyze the association between in-patient cost and variables at patient, hospital, and state levels. Results: This study analyzed data on 9066 DLEA hospitalizations from 39 states. The mean cost per in-patient stay was $17,103. Four out of the five most costly states were located on the East and West coasts (NY and NJ, CA and OR). Age, race, length of stay, level of amputation, in-patient mortality, primary payer, co-morbidities, and type of hospital were significantly correlated with in-patient costs and explained 55.3% of the cost variance. Based on the means of costs unexplained by those factors, the three West coast states had the highest costs, followed by five Midwestern states, and four Southern states, and Kansas were the least costly. Conclusions: Over 40% of the variations in DLEA hospital costs could not be explained by major patient-, hospital-, and state-level variables. Further research is needed to examine whether similar patterns exist for other costly surgical procedures among diabetic patients. © 2013 Informa UK Ltd. All rights reserved: reproduction in whole or part not permitted.

Michelson D.,Merck And Co. | Snyder E.,Merck And Co. | Paradis E.,Merck And Co. | Chengan-Liu M.,Merck And Co. | And 9 more authors.
The Lancet Neurology | Year: 2014

Background: Suvorexant (MK-4305) is an orexin receptor antagonist shown to be efficacious for insomnia over 3 months. We aimed to assess its clinical profile during and after 1 year of treatment. Methods: We did a randomised, placebo-controlled, parallel-group trial at 106 investigational centres in the Americas, Australia, Europe, and South Africa from December, 2009, to August, 2011. Patients aged 18 years or older with primary insomnia by DSM-IV-TR criteria were assigned using a computer-generated randomised allocation schedule to receive nightly suvorexant (40 mg for patients younger than 65 years, 30 mg for patients aged 65 years or older) or placebo at a 2:1 ratio for 1 year with a subsequent 2-month randomised discontinuation phase in which patients on suvorexant either continued suvorexant or were abruptly switched to placebo while patients on placebo remained on placebo. Treatment assignment was masked from patients and investigators. The primary objective was to assess the safety and tolerability of suvorexant for up to 1 year. Secondary objectives were to assess the efficacy of suvorexant for improving patient-reported subjective total sleep time (sTST) and time to sleep onset (sTSO) over the first month of treatment. Efficacy endpoints over the first month were assessed with a mixed model with terms for baseline value of the response variable, age, sex, region, treatment, time, and treatment by time interaction. This trial is registered with ClinicalTrials.gov, number NCT01021813. Findings: 322 (62%) of 522 patients randomly assigned to receive suvorexant and 162 (63%) of 259 assigned to receive placebo completed the 1-year phase. Over 1 year, 362 (69%) of 521 patients treated with suvorexant experienced any adverse events compared with 164 (64%) of 258 treated with placebo. Serious adverse events were recorded in 27 patients (5%) who received suvorexant and 17 (7%) who received placebo. The most common adverse event, somnolence, was reported for 69 patients (13%) who received suvorexant and seven (3%) who received placebo. At month 1, suvorexant (517 patients in the efficacy population) showed greater efficacy than placebo (254 in the efficacy population) in improving sTST (38·7 min vs 16·0 min; difference 22·7, 95% CI 16·4 to 29·0; p<0·0001) and sTSO (-18·0 min vs -8·4 min, difference -9·5, -14·6 to -4·5; p=0·0002). Interpretation: Our findings show that suvorexant was generally safe and well tolerated over 1 year of nightly treatment in patients with insomnia, with efficacy noted for subjective measures of sleep onset and maintenance. Funding: Merck & Co Inc. © 2014 Elsevier Ltd.

Agency: European Commission | Branch: H2020 | Program: IA | Phase: NMP-04-2014 | Award Amount: 7.88M | Year: 2015

Bringing intelligence and communication to everyday objects is a major challenge for future electronics. This Internet of Things concept envisions wide dissemination with new performances: robustness, large area, flexibility, eco-efficient large volume manufacturing at low cost. Beyond current TOLAE demonstration, a major technology jump driving the scalability towards nanoscale resolution via high-definition cost-effective printing is required to deliver the properties and electrical performances expected by applications. ATLASS Innovation Action takes this huge step by bringing high resolution technologies to the printing industries for the demonstration of products at TRL6 in high impact markets. New multifunctional high-performing inks (semiconductor mobility >1cm2/Vs, dielectrics, ferroelectrics) and high-resolution (down to 500nm and ~100nm thickness) R2R/S2S printing including nano-imprinting and gravure printing will be engineered and scaled-up on pre-industrial pilot lines, enabling high performance devices (speed ~ 10 MHz). In-line control and novel automatic optical inspection tools and methodology will be installed to ramp-up the yield of developed processes (>99%) thus enabling cost-efficient fabrication of advanced circuits (>1000 transistors, 50kHz clock rate). The technology capability is benchmarked with conventional TOLAE process and demonstrated with 4 applications in the field of Interactive objects and Sensing surfaces (temperature tag for smart food packaging, electronic label for logistics, impact force sensing foils for automotive safety -, proximity sensing for safer human-robot collaboration ). With a consortium of 11 top companies (7 SMEs) from the cutting-edge, fast growing printed electronics sector and 4 RTOs with high-level technology expertise, ATLASS will strongly impact the global market of sensors, labels and smart objects expected to reach revenue of several EUR billion with printed sensors share of EUR 644 million by 2022.

Agency: European Commission | Branch: H2020 | Program: MSCA-ITN-ETN | Phase: MSCA-ITN-2016 | Award Amount: 3.83M | Year: 2016

Organic solar cells (OSCs) have the potential to become an environmental friendly, inexpensive, large area and flexible photovoltaics technology. Their main advantages are low process temperatures, the potential for very low cost due to abundant materials and scalable processing, and the possibility of producing flexible devices on plastic substrates. To improve their commercialization capacity, to compete with established power generation and to complement other renewable energy technologies, the performance of state-of-the-art OSCs needs to be further improved. Our goals within SEPOMO Spins in Efficient Photovoltaic devices based on Organic Molecules are to bring the performance of OSCs forward by taking advantage of the so far unexplored degree of freedom of photogenerated species in organic materials, their spin. This challenging idea provides a unified platform for the excellent research to promote the world-wide position of Europe in the field of organic photovoltaics and electronics, and to train strongly motivated early stage researchers (ESRs) for a career in science and technology oriented industry that is rapidly growing. Our scientific objectives are to develop several novel routes to enhance the efficiency of OSC by understanding and exploiting the electronic spin interactions. This will allow us to address crucial bottlenecks in state-of-the-art OSCs: we will increase the quantum efficiency by reducing the dominant recombination losses and by enhancing the light harvesting and exciton generation, e.g. by means of internal upconversion of excited states. Our ESRs will be trained within this interdisciplinary (physics, chemistry, engineering) and intersectoral (academia, R&D center, enterprise) consortium in highly relevant fundamental yet application-oriented research with the potential to commercialise the results. The hard and soft skills learned in our network are central for the ESRs to pursue their individual careers in academics or industry.

Kastelein J.J.P.,University of Amsterdam | Besseling J.,University of Amsterdam | Shah S.,Merck And Co. | Bergeron J.,Laval University | And 8 more authors.
The Lancet | Year: 2015

Summary Background Present guidelines emphasise the importance of low concentrations of LDL cholesterol (LDL-C) in patients with familial hypercholesterolaemia. In most patients with the disease, however, these concentrations are not achieved with present treatments, so additional treatment is therefore warranted. Inhibition of cholesteryl ester transfer protein has been shown to reduce LDL-C concentrations in addition to regular statin treatment in patients with hypercholesterolaemia or at high risk of cardiovascular disease. We aimed to investigate the safety and efficacy of anacetrapib, a cholesteryl ester transfer protein inhibitor, in patients with heterozygous familial hypercholesterolaemia. Methods In this multicentre, randomised, double-blind, placebo-controlled, phase 3 study, patients aged 18-80 years with a genotype-confirmed or clinical diagnosis of heterozygous familial hypercholesterolaemia, on optimum lipid-lowering treatment for at least 6 weeks, and with an LDL-C concentration of 2·59 mmol/L or higher without cardiovascular disease or 1·81 mmol/L or higher with cardiovascular disease from 26 lipid clinics across nine countries were eligible. We randomly allocated participants with a computer-generated allocation schedule (2:1; block size of six; no stratification) to oral anacetrapib 100 mg or placebo for 52 weeks, with a 12 week post-treatment follow-up afterwards. We masked patients, care providers, and those assessing outcomes to treatment groups throughout the study. The primary outcome was percentage change from baseline in LDL-C concentration. We did analysis using a constrained longitudinal repeated measures model. This trial is registered with ClinicalTrials.gov, number NCT01524289. Findings Between Feb 10, 2012, and Feb 12, 2014, we randomly allocated 204 patients to anacetrapib and 102 to placebo. One patient in the anacetrapib group did not receive the drug. At week 52, anacetrapib reduced mean LDL-C concentration from 3·3 mmol/L (SD 0·8) to 2·1 mmol/L (0·8; percentage change 36·0% [95% CI -39·5 to -32·5] compared with an increase with placebo from 3·4 mmol/L (1·2) to 3·5 mmol/L (1·6; percentage change 3·7% [-1·2 to 8·6], with a difference in percentage change between anacetrapib and placebo of -39·7% (95% CI -45·7 to -33·7; p<0·0001). The number of cardiovascular events was increased in patients given anacetrapib compared with those given placebo (4 [2%] of 203 vs none [0%] of 102; p=0·1544), but the proportion with adverse events leading to discontinuation was similar (12 [6%] of 203 vs five [5%] of 102). Interpretation In patients with heterozygous familial hypercholesterolaemia, treatment with anacetrapib for 1 year was well tolerated and resulted in substantial reductions in LDL-C concentration. Whether this change leads to a reduction of cardiovascular events will be answered in an outcome study. Funding Merck & Co, Inc © 2015 Elsevier Ltd.

Gibson T.B.,Thomson Reuters | Mahoney J.,Florida Health Care Coalition | Ranghell K.,Florida Health Care Coalition | Cherney B.J.,Florida Health Care Coalition | McElwee N.,Merck And Co.
Health Affairs | Year: 2011

We evaluated the effects of implementing a value-based insurance design program for patients with diabetes in two groups within a single firm. One group participated in disease management; the other did not. We matched members of the two groups to similar enrollees within the company that did not offer the value-based program. We found that participation in both value-based insurance design and disease management resulted in sustained improvement over time. Use of diabetes medications increased 6.5 percent over three years. Adherence to diabetes medical guidelines also increased, producing a return on investment of $1.33 saved for every dollar spent during a three-year follow-up period. ©2011 Project HOPE - The People-to-People Health Foundation, Inc.

Lopez D.,Autonomous University of Madrid | Garcia-Calvo M.,Merck And Co. | Smith G.L.,Imperial College London | Del Val M.,Autonomous University of Madrid
Journal of Immunology | Year: 2010

CD8+ cytotoxic T lymphocytes recognize infected cells in which MHC class I molecules present pathogen-derived peptides that have been processed mainly by proteasomes. Many infections induce a set of proteases, the caspases involved in apoptosis or inflammation. In this study, we report that processing and presentation of a short vaccinia virus-encoded Ag can take place also by a nonproteasomal pathway, which was blocked in infected cells with chemical inhibitors of caspases. By cleaving at noncanonical sites, at least two caspases generated antigenic peptides recognized by T lymphocytes. The sites and the peptidic products were partially overlapping but different to those used and produced by proteasomes in vitro. Antigenic natural peptides produced in infected cells by either pathway were quantitatively and qualitatively similar. Finally, coexpression of the natural vaccinia virus protein B13, which is an inhibitor of caspases and apoptosis, impaired Ag presentation by the caspase pathway in infected cells. These data support the hypothesis that numerous cellular proteolytic systems, including those induced during infection, such as caspases involved in apoptosis or in inflammation, contribute to the repertoire of presented peptides, thereby facilitating immunosurveillance. Copyright © 2010 by The American Association of Immunologists, Inc.

Black D.M.,University of California at San Francisco | Kelly M.P.,University of California at San Francisco | Genant H.K.,University of California at San Francisco | Palermo L.,University of California at San Francisco | And 8 more authors.
New England Journal of Medicine | Year: 2010

Background: A number of recent case reports and series have identified a subgroup of atypical fractures of the femoral shaft associated with bisphosphonate use. A population-based study did not support this association. Such a relationship has not been examined in randomized trials. Methods: We performed secondary analyses using the results of three large, randomized bisphosphonate trials: the Fracture Intervention Trial (FIT), the FIT Long-Term Extension (FLEX) trial, and the Health Outcomes and Reduced Incidence with Zoledronic Acid Once Yearly (HORIZON) Pivotal Fracture Trial (PFT). We reviewed fracture records and radiographs (when available) from all hip and femur fractures to identify those below the lesser trochanter and above the distal metaphyseal flare (subtrochanteric and diaphyseal femur fractures) and to assess atypical features. We calculated the relative hazards for subtrochanteric and diaphyseal fractures for each study. Results: We reviewed 284 records for hip or femur fractures among 14,195 women in these trials. A total of 12 fractures in 10 patients were classified as occurring in the subtrochanteric or diaphyseal femur, a combined rate of 2.3 per 10,000 patient-years. As compared with placebo, the relative hazard was 1.03 (95% confidence interval [CI], 0.06 to 16.46) for alendronate use in the FIT trial, 1.50 (95% CI, 0.25 to 9.00) for zoledronic acid use in the HORIZON-PFT trial, and 1.33 (95% CI, 0.12 to 14.67) for continued alendronate use in the FLEX trial. Although increases in risk were not significant, confidence intervals were wide. Conclusions: The occurrence of fracture of the subtrochanteric or diaphyseal femur was very rare, even among women who had been treated with bisphosphonates for as long as 10 years. There was no significant increase in risk associated with bisphosphonate use, but the study was underpowered for definitive conclusions. Copyright © 2010 Massachusetts Medical Society.

Burghardt A.J.,University of California at San Francisco | Kazakia G.J.,University of California at San Francisco | Sode M.,University of California at San Francisco | Sode M.,University of California at Berkeley | And 4 more authors.
Journal of Bone and Mineral Research | Year: 2010

The goal of this study was to characterize longitudinal changes in bone microarchitecture and function in women treated with an established antifracture therapeutic. In this double-blind, placebo-controlled pilot study, 53 early postmenopausal women with low bone density (age=56±4 years; femoral neck T-score=-1.5±0.6) were monitored by high-resolution peripheral quantitative computed tomography (HR-pQCT) for 24 months following randomization to alendronate (ALN) or placebo (PBO) treatment groups. Subjects underwent annual HR-pQCT imaging of the distal radius and tibia, dual-energy X-ray absorptiometry (DXA), and determination of biochemical markers of bone turnover (BSAP and uNTx). In addition to bone density and microarchitecture assessment, regional analysis, cortical porosity quantification, and micro-finite-element analysis were performed. After 24 months of treatment, at the distal tibia but not the radius, HR-pQCT measures showed significant improvements over baseline in the ALN group, particularly densitometric measures in the cortical and trabecular compartments and endocortical geometry (cortical thickness and area, medullary area) (p<.05). Cortical volumetric bone mineral density (vBMD) in the tibia alone showed a significant difference between treatment groups after 24 months (p<.05); however, regionally, significant differences in Tb.vBMD, Tb.N, and Ct.Th were found for the lateral quadrant of the radius (p<.05). Spearman correlation analysis revealed that the biomechanical response to ALN in the radius and tibia was specifically associated with changes in trabecular microarchitecture (|ρ|=0.51 to 0.80, p<.05), whereas PBO progression of bone loss was associated with a broad range of changes in density, geometry, and microarchitecture (|ρ|=0.56 to 0.89, p<.05). Baseline cortical geometry and porosity measures best predicted ALN-induced change in biomechanics at both sites (ρ > 0.48, p<.05). These findings suggest a more pronounced response to ALN in the tibia than in the radius, driven by trabecular and endocortical changes. © 2010 American Society for Bone and Mineral Research.

Gibertini M.,INC Research | Nations K.R.,Merck And Co. | Whitaker J.A.,INC Research
International Clinical Psychopharmacology | Year: 2012

The high failure rate of antidepressant trials has spurred exploration of the factors that affect trial sensitivity. In the current analysis, Food and Drug Administration antidepressant drug registration trial data compiled by Turner et al. is extended to include the most recently approved antidepressants. The expanded dataset is examined to further establish the likely population effect size (ES) for monoaminergic antidepressants and to demonstrate the relationship between observed ES and sample size in trials on compounds with proven efficacy. Results indicate that the overall underlying ES for antidepressants is approximately 0.30, and that the variability in observed ES across trials is related to the sample size of the trial. The current data provide a unique real-world illustration of an often underappreciated statistical truism: that small N trials are more likely to mislead than to inform, and that by aligning sample size to the population ES, risks of both erroneously high and low effects are minimized. The results in the current study make this abstract concept concrete and will help drug developers arrive at informed gate decisions with greater confidence and fewer risks, improving the odds of success for future antidepressant trials. © 2012 Wolters Kluwer Health | Lippincott Williams & Wilkins.

Narayanan M.,Seattle Genetics | Narayanan M.,Merck And Co. | Vetta A.,McGill University | Schadt E.E.,Seattle Genetics | And 2 more authors.
PLoS Computational Biology | Year: 2010

Many genome-wide datasets are routinely generated to study different aspects of biological systems, but integrating them to obtain a coherent view of the underlying biology remains a challenge. We propose simultaneous clustering of multiple networks as a framework to integrate large-scale datasets on the interactions among and activities of cellular components. Specifically, we develop an algorithm JointCluster that finds sets of genes that cluster well in multiple networks of interest, such as coexpression networks summarizing correlations among the expression profiles of genes and physical networks describing protein-protein and protein-DNA interactions among genes or gene-products. Our algorithm provides an efficient solution to a well-defined problem of jointly clustering networks, using techniques that permit certain theoretical guarantees on the quality of the detected clustering relative to the optimal clustering. These guarantees coupled with an effective scaling heuristic and the flexibility to handle multiple heterogeneous networks make our method JointCluster an advance over earlier approaches. Simulation results showed JointCluster to be more robust than alternate methods in recovering clusters implanted in networks with high false positive rates. In systematic evaluation of JointCluster and some earlier approaches for combined analysis of the yeast physical network and two gene expression datasets under glucose and ethanol growth conditions, JointCluster discovers clusters that are more consistently enriched for various reference classes capturing different aspects of yeast biology or yield better coverage of the analysed genes. These robust clusters, which are supported across multiple genomic datasets and diverse reference classes, agree with known biology of yeast under these growth conditions, elucidate the genetic control of coordinated transcription, and enable functional predictions for a number of uncharacterized genes. © 2010 Narayanan et al.

Hall A.,University of Pennsylvania | Beiting D.,University of Pennsylvania | Tato C.,Merck And Co. | John B.,University of Pennsylvania | And 19 more authors.
Immunity | Year: 2012

Interferon-γ (IFN-γ) promotes a population of T-bet+ CXCR3+ regulatory T (Treg) cells that limit T helper 1 (Th1) cell-mediated pathology. Our studies demonstrate that interleukin-27 (IL-27) also promoted expression of T-bet and CXCR3 in Treg cells. During infection with Toxoplasma gondii, a similar population emerged that limited T cell responses and was dependent on IFN-γ in the periphery but on IL-27 at mucosal sites. Transfer of Treg cells ameliorated the infection-induced pathology observed in Il27-/- mice, and this was dependent on their ability to produce IL-10. Microarray analysis revealed that Treg cells exposed to either IFN-γ or IL-27 have distinct transcriptional profiles. Thus, IFN-γ and IL-27 have different roles in Treg cell biology and IL-27 is a key cytokine that promotes the development of Treg cells specialized to control Th1 cell-mediated immunity at local sites of inflammation. © 2012 Elsevier Inc.

Pfaller M.A.,University of Iowa | Diekema D.J.,University of Iowa | Andes D.,University of Wisconsin - Madison | Arendrup M.C.,Statens Serum Institute | And 4 more authors.
Drug Resistance Updates | Year: 2011

The CLSI established clinical breakpoints (CBPs) for caspofungin (CSF), micafungin (MCF) and anidulafungin (ANF) versus Candida. The same CBP (susceptible (S): MIC ≤ 2 mcg/ml; non-S: MIC > 2 mcg/ml) was applied to all echinocandins and species. More data now allow reassessment of these CBPs. We examined cases of echinocandin failure where both MICs and fks mutations were assessed; wild type (WT) MICs and epidemiological cutoff values (ECVs) for a large Candida collection; molecular analysis of fks hotspots for Candida with known MICs; and pharmacokinetic and pharmacodynamic (PK/PD) data. We applied these findings to propose new species-specific CBPs for echinocandins and Candida. Of 18 candidiasis cases refractory to echinocandins and with fks mutations, 28% (CSF), 58% (ANF) and 66% (MCF) had MICs in the S category using CBP of ≤2 mcg/ml, while 0-8% would be S using CBP of ≤0.25 mcg/ml. WT MIC distributions revealed ECV ranges of 0.03-0.25 mcg/ml for all major species except C. parapsilosis (1-4 mcg/ml) and C. guilliermondii (4-16 mcg/ml). Among Candida tested for fks mutations, only 15.7-45.1% of 51 mutants were detected using the CBP for NS of >2 mcg/ml. In contrast, a cutoff of >0.25 mcg/ml for C. albicans, C. tropicalis, C. krusei, and C. dubliniensis detected 85.6% (MCF) to 95.2% (CSF) of 21 mutant strains. Likewise, a cutoff of >0.12 mcg/ml for ANF and CSF and of >0.06 mcg/ml for MCF detected 93% (ANF) to 97% (CSF, MCF) of 30 mutant strains of C. glabrata. These data, combined with PK/PD considerations, support CBPs of ≤0.25 mcg/ml (S), 0.5 mcg/ml (I), ≥1 (R) for CSF/MCF/ANF and C. albicans, C. tropicalis and C. krusei and ≤2 mcg/ml (S), 4 mcg/ml (I), and ≥8 mcg/ml (R) for these agents and C. parapsilosis. The CBPs for ANF and CSF and C. glabrata are ≤0.12 mcg/ml (S), 0.25 mcg/ml (I), and ≥0.5 mcg/ml (R), whereas those for MCF are ≤0.06 mcg/ml (S), 0.12 mcg/ml (I), and ≥0.25 mcg/ml (R). New, species-specific CBPs for Candida and the echinocandins are more sensitive to detect emerging resistance associated with fks mutations, and better able to predict risk for clinical failure. © 2011 Elsevier Ltd. All rights reserved.

Murthi A.,Ohio State University | Murthi A.,National Institute of Allergy and Infectious Diseases | Shaheen H.H.,Ohio State University | Shaheen H.H.,Merck And Co. | And 5 more authors.
Molecular Biology of the Cell | Year: 2010

tRNAs in yeast and vertebrate cells move bidirectionally and reversibly between the nucleus and the cytoplasm. We investigated roles of members of the β-importin family in tRNA subcellular dynamics. Retrograde import of tRNA into the nucleus is dependent, directly or indirectly, upon Mtr10. tRNA nuclear export utilizes at least two members of the β-importin family. The β-importins involved in nuclear export have shared and exclusive functions. Los1 functions in both the tRNA primary export and the tRNA reexport processes. Msn5 is unable to export tRNAs in the primary round of export if the tRNAs are encoded by intron-containing genes, and for these tRNAs Msn5 functions primarily in their reexport to the cytoplasm. The data support a model in which tRNA retrograde import to the nucleus is a constitutive process; in contrast, reexport of the imported tRNAs back to the cytoplasm is regulated by the availability of nutrients to cells and by tRNA aminoacylation in the nucleus. Finally, we implicate Tef1, the yeast orthologue of translation elongation factor eEF1A, in the tRNA reexport process and show that its subcellular distribution between the nucleus and cytoplasm is dependent upon Mtr10 and Msn5. © 2010 by The American Society for Cell Biology.

Wagner C.,Goethe University Frankfurt | Jantratid E.,Goethe University Frankfurt | Kesisoglou F.,Merck And Co. | Vertzoni M.,National and Kapodistrian University of Athens | And 2 more authors.
European Journal of Pharmaceutics and Biopharmaceutics | Year: 2012

For predicting food effects and simulating plasma profiles of poorly soluble drugs, physiologically based pharmacokinetic models have become a widely accepted tool in academia and the pharmaceutical industry. Up till now, however, simulations appearing in the open literature have mainly focused on BCS class II compounds, and many of these simulations tend to have more of a "retrospective" than a prognostic, predictive character. In this work, investigations on the absorption of a weakly basic BCS class IV drug, "Compound A", were performed. The objective was to predict the plasma profiles of an immediate release (IR) formulation of Compound A in the fasted and fed state. For this purpose, in vitro biorelevant dissolution tests and transfer model experiments were conducted. Dissolution and precipitation kinetics were then combined with in vivo post-absorptive disposition parameters using STELLA® software. As Compound A not only exhibits poor solubility but also poor permeability, a previously developed STELLA® model was revised to accommodate the less than optimal permeability characteristics as well as precipitation of the drug in the fasted state small intestine. Permeability restrictions were introduced into the model using an absorption rate constant calculated from the Caco-2 permeability value of Compound A, the effective intestinal surface area and appropriate intestinal fluid volumes. The results show that biorelevant dissolution tests are a helpful tool to predict food effects of Compound A qualitatively. However, the plasma profiles of Compound A could only be predicted quantitatively when the results of biorelevant dissolution test were coupled with the newly developed PBPK model. © 2012 Elsevier B.V. All rights reserved.

Faith M.S.,University of Pennsylvania | Butryn M.,Drexel University | Wadden T.A.,University of Pennsylvania | Fabricatore A.,University of Pennsylvania | And 2 more authors.
Obesity Reviews | Year: 2011

Obesity may lead to depression or be one of its consequences. We reviewed population-based studies in order to, first, identify the most commonly used research methods, and, second, to evaluate the strength of evidence for prospective associations among obesity and depression. We examined 25 studies, of which 10 tested 'obesity-to-depression' pathways, and 15 tested 'depression-to-obesity' pathways. Descriptive statistics summarized the frequency with which various measurements, designs and data analytic strategies were used. We tallied the number of studies that reported any vs. no statistically significant associations, and report on effect sizes, identified moderating variables within reports, and sought common findings across studies. Results indicated considerable methodological heterogeneity in the literature. Depression was assessed by clinical interview in 44% of studies, weight and height were directly measured in 32%, and only 12% used both. In total, 80% of the studies reported significant obesity-to-depression associations, with odds ratios generally in the range of 1.0 to 2.0, while only 53% of the studies reported significant depression-to-obesity associations. Sex was a common moderating variable. Thus, there was good evidence that obesity is prospectively associated with increased depression, with less consistent evidence that depression leads to obesity. Recommendations for future research regarding study samples, measurement and data analysis are provided. © 2011 The Authors. obesity reviews © 2011 International Association for the Study of Obesity.

Young R.M.,University of Pennsylvania | Ackerman D.,University of Pennsylvania | Quinn Z.L.,University of Pennsylvania | Quinn Z.L.,Howard Hughes Medical Institute | And 14 more authors.
Genes and Development | Year: 2013

Solid tumors exhibit heterogeneous microenvironments, often characterized by limiting concentrations of oxygen (O2), glucose, and other nutrients. How oncogenic mutations alter stress response pathways, metabolism, and cell survival in the face of these challenges is incompletely understood. Here we report that constitutive mammalian target of rapamycin complex 1 (mTORC1) activity renders hypoxic cells dependent on exogenous desaturated lipids, as levels of de novo synthesized unsaturated fatty acids are reduced under low O2. Specifically, we demonstrate that hypoxic Tsc2-/- (tuberous sclerosis complex 2-/-) cells deprived of serum lipids exhibit a magnified unfolded protein response (UPR) but fail to appropriately expand their endoplasmic reticulum (ER), leading to inositol-requiring protein-1 (IRE1)-dependent cell death that can be reversed by the addition of unsaturated lipids. UPR activation and apoptosis were also detected in Tsc2-deficient kidney tumors. Importantly, we observed this phenotype in multiple human cancer cell lines and suggest that cells committed to unregulated growth within ischemic tumor microenvironments are unable to balance lipid and protein synthesis due to a critical limitation in desaturated lipids. © 2013 by Cold Spring Harbor Laboratory Press.

Ho T.W.,Merck And Co. | Edvinsson L.,Lund University | Goadsby P.J.,University of California at San Francisco
Nature Reviews Neurology | Year: 2010

Over the past 300 years, the migraine field has been dominated by two main theories-the vascular theory and the central neuronal theory. The success of vasoconstrictors such as ergotamine and the triptans in treating acute migraine bolstered the vascular theory, but evidence is now emerging that vasodilatation is neither necessary nor sufficient to induce a migraine attack. Attention is now turning to the core migraine circuits in the brain, which include the trigeminal ganglia, trigeminal nucleus, medullary modulatory regions, pons, periaqueductal gray matter, hypothalamus and thalamus. Migraine triggers are likely to reflect a disturbance in overall balance of the circuits involved in the modulation of sensory activity, particularly those with relevance to the head. In this Review, we consider the evidence pointing towards a neuronal mechanism in migraine development, highlighting the role of calcitonin gene-related peptide (CGRP), which is found in small to medium-sized neurons in the trigeminal ganglion. CGRP is released during migraine attacks and can trigger migraine in patients, and CGRP receptor antagonists can abort migraine. We also examine whether other drugs, such as triptans, might exert their antimigraine effects via their actions on the neuronal circuit as opposed to the intracranial vasculature. © 2010 Macmillan Publishers Limited. All rights reserved.

Katlama C.,University Pierre and Marie Curie | Deeks S.G.,University of California at San Francisco | Autran B.,University Pierre and Marie Curie | Martinez-Picado J.,Autonomous University of Barcelona | And 8 more authors.
The Lancet | Year: 2013

Antiretroviral therapy for HIV infection needs lifelong access and strict adherence to regimens that are both expensive and associated with toxic effects. A curative intervention will be needed to fully stop the epidemic. The failure to eradicate HIV infection during long-term antiretroviral therapy shows the intrinsic stability of the viral genome in latently infected CD4T cells and other cells, and possibly a sustained low-level viral replication. Heterogeneity in latently infected cell populations and homoeostatic proliferation of infected cells might affect the dynamics of virus production and persistence. Despite potent antiretroviral therapy, chronic immune activation, inflammation, and immune dysfunction persist, and are likely to have important effects on the size and distribution of the viral reservoir. The inability of the immune system to recognise cells harbouring latent virus and to eliminate cells actively producing virus is the biggest challenge to finding a cure. We look at new approaches to unravelling the complex virus-host interactions that lead to persistent infection and latency, and discuss the rationale for combination of novel treatment strategies with available antiretroviral treatment options to cure HIV. Copyright © 2013 Elsevier B.V.

Kendrick J.,University of Colorado at Denver | Shlipak M.G.,University of California at San Francisco | Targher G.,University of Verona | Cook T.,Merck And Co. | And 2 more authors.
American Journal of Kidney Diseases | Year: 2010

Background: Chronic kidney disease (CKD) is associated with an increased risk of incident cardiovascular disease (CVD); however, the role of statins for the primary prevention of acute cardiovascular events in patients with CKD and the effect of statins on kidney function loss in persons without prevalent CVD have not been studied. Study Design: Post hoc analysis of the Air Force/Texas Coronary Atherosclerosis Prevention Study. Setting & Participants: Multicenter, randomized, double-blind, placebo-controlled trial of 5,608 men and 997 women without CVD randomly assigned to treatment with lovastatin or placebo. Intervention: Placebo or lovastatin, 20 mg/d. Outcomes & Measurements: First major acute cardiovascular event in participants with mild CKD and kidney function loss in persons with or without CKD. Estimated glomerular filtration rate was calculated using the 4-variable Modification of Diet in Renal Disease Study equation. Results: At baseline, mean estimated glomerular filtration rate in participants with CKD (n = 304) was 53.0 ± 6.0 mL/min/1.73 m2. After an average follow-up of 5.3 ± 0.8 years, the incidence of a fatal and nonfatal CVD event was lower in participants with CKD receiving lovastatin than in those receiving placebo (adjusted relative risk [RR], 0.31; 95% CI, 0.13-0.72; P = 0.01). Tests for interaction suggested that the benefit of lovastatin was independent of the presence of CKD. Lovastatin did not reduce the annualized mean decrease in estimated glomerular filtration rate (-1.3 ± 0.07 vs -1.4 ± 0.07 mL/min/1.73 m2/y, respectively; P = 0.1) or the frequency of a ≥ 25% decrease in kidney function (adjusted RR, 1.10; 95% CI, 0.96-1.28; P = 0.2) or incident CKD (adjusted RR, 1.04; 95% CI, 0.86-1.27; P = 0.6). Limitations: Unable to determine the cause and duration of kidney disease, and information regarding proteinuria was not available. Conclusions: Lovastatin is effective for the primary prevention of CVD in patients with CKD, but is not effective in decreasing kidney function loss in persons with no CVD. © 2009 National Kidney Foundation, Inc.

Roth D.,University of Miami | Nelson D.R.,Clinical Translational Science Institute | Bruchfeld A.,Karolinska University Hospital | Liapakis A.,Yale University | And 15 more authors.
The Lancet | Year: 2015

Summary Background Chronic hepatitis C virus (HCV) infection in patients with stage 4-5 chronic kidney disease increases the risk of death and renal graft failure, yet patients with hepatitis C and chronic kidney disease have few treatment options. This study assesses an all-oral, ribavirin-free regimen in patients with HCV genotype 1 infection and stage 4-5 chronic kidney disease. Methods In this phase 3 randomised study of safety and observational study of efficacy, patients with HCV genotype 1 infection and chronic kidney disease (stage 4-5 with or without haemodialysis dependence) were randomly assigned to receive grazoprevir (100 mg, NS3/4A protease inhibitor) and elbasvir (50 mg, NS5A inhibitor; immediate treatment group) or placebo (deferred treatment group) once daily for 12 weeks. Randomisation was done centrally with an interactive voice response system. An additional cohort of patients who were not randomised received the same regimen open-label and underwent intensive pharmacokinetic sampling. The primary efficacy outcome was a non-randomised comparison of sustained virological response at 12 weeks (SVR12) after the end of therapy for the combined immediate treatment group and the pharmacokinetic population with a historical control. The primary safety outcome was a randomised comparison between the immediate treatment group and the deferred treatment group. After 4 weeks of follow-up (study week 16), unmasking occurred and patients in the deferred treatment group received grazoprevir and elbasvir. The primary efficacy hypothesis was tested at a two-sided significance level (type I error) of 0·05 using an exact test for a binomial proportion. Safety event rates were compared between immediate treatment and deferred treatment groups using the stratified Miettinen and Nurminen method with baseline dialysis status as the strata. The study is registered at ClinicalTrials.gov, number NCT02092350. Findings 224 patients were randomly assigned to the immediate treatment group with grazoprevir and elbasvir (n=111) or the deferred treatment group (n=113), and 11 were assigned to the intensive pharmacokinetic population. Overall, 179 (76%) were haemodialysis-dependent, 122 (52%) had HCV genotype 1a infection, 189 (80%) were HCV treatment-naive, 14 (6%) were cirrhotic, and 108 (46%) were African American. Of the 122 patients receiving grazoprevir and elbasvir, six were excluded from the primary efficacy analysis for non-virological reasons (death, lost-to-follow-up [n=2], non-compliance, patient withdrawal, and withdrawal by physician for violent behaviour). No patients in the combined immediate treatment group and intensive pharmacokinetic population and five (4%) in the deferred treatment group discontinued because of an adverse event. Most common adverse events were headache, nausea, and fatigue, occurring at similar frequencies in patients receiving active and placebo drugs. SVR12 in the combined immediate treatment group and intensive pharmacokinetic population was 99% (95% CI 95·3-100·0; 115/116), with one relapse 12 weeks after end of treatment when compared with a historical control of 45%, based on meta-analyses of interferon-based regimens used in clinical trials of patients infected with HCV who are on haemodialysis. Interpretation Once-daily grazoprevir and elbasvir for 12 weeks had a low rate of adverse events and was effective in patients infected with HCV genotype 1 and stage 4-5 chronic kidney disease. Funding Merck Sharp & Dohme Corp. © 2015 Elsevier Ltd.

Girman C.J.,Merck And Co. | Kou T.D.,Merck And Co. | Cai B.,Merck And Co. | Alexander C.M.,Global Medical Affairs | And 3 more authors.
Diabetes, Obesity and Metabolism | Year: 2010

Aim: The aetiology of acute pancreatitis (AP) is complex, and many risk factors for AP are shared by patients with type 2 diabetes mellitus (T2DM). However, few have assessed risk factors for AP specifically in T2DM patients.Methods: Patients in the General Practice Research Database (2 984 755, 5.0% with T2DM) were used to estimate incidence of AP for T2DM relative to non-diabetes, adjusting for prior pancreatitis, gallbladder disease, obesity, smoking and alcohol use. Multivariate Cox regression analysis adjusting for risk factors and Charlson comorbidity index (CCI) was used to estimate hazard ratios (HR) with 95% confidence intervals (CI).Results: Between 2003 and 2007, 301 of 148 903 patients with T2DM and 2434 of almost 3 million patients without diabetes developed AP. Patients with T2DM had higher risk for AP compared with patients without diabetes (crude HR: 2.89, 95% CI: 2.56-3.27). Patients with T2DM had significantly higher rates of prior alcohol and tobacco exposure (44.2 and 61.9% vs. 34.1 and 35.9%, p < 0.001) and of comorbid conditions (14.7% with CCI ≥1 vs. 4.3%, p < 0.001). Histories of obesity, pancreatitis, gallbladder disease, smoking or alcohol use were significant predictors of AP. After adjusting for these factors, age, gender and comorbidities, the risk of developing AP remained elevated in patients with T2DM (adjusted HR: 1.49, 95% CI: 1.31-1.70).Conclusion: After adjusting for risk factors, patients with T2DM had an elevated risk of AP compared with patients without diabetes. Physicians should be aware of the increased risk in patients with T2DM, particularly in those with prior pancreatitis. © 2010 Blackwell Publishing Ltd.

Jensen P.R.,University of Southern Denmark | Jensen P.R.,Vejle Hospital | Andersen T.L.,University of Southern Denmark | Pennypacker B.L.,Merck And Co. | And 2 more authors.
Calcified Tissue International | Year: 2014

Odanacatib (ODN) is a bone resorption inhibitor which differs from standard antiresorptives by its ability to reduce bone resorption without decreasing bone formation. What is the reason for this difference? In contrast with other antiresorptives, such as alendronate (ALN), ODN targets only the very last step of the resorption process. We hypothesize that ODN may therefore modify the remodeling events immediately following osteoclastic resorption. These events belong to the reversal phase and include recruitment of osteoblasts, which is critical for connecting bone resorption to formation. We performed a histomorphometric study of trabecular remodeling in vertebrae of estrogen-deficient rabbits treated or not with ODN or ALN, a model where ODN, but not ALN, was previously shown to preserve bone formation. In line with our hypothesis, we found that ODN treatment compared to ALN results in a shorter reversal phase, faster initiation of osteoid deposition on the eroded surfaces, and higher osteoblast recruitment. The latter is reflected by higher densities of mature bone forming osteoblasts and an increased subpopulation of cuboidal osteoblasts. Furthermore, we found an increase in the interface between osteoclasts and surrounding osteoblast-lineage cells. This increase is expected to favor the osteoclast-osteoblast interactions required for bone formation. Regarding bone resorption itself, we show that ODN, but not ALN, treatment results in shallower resorption lacunae, a geometry favoring bone stiffness. We conclude that, compared to standard antiresorptives, ODN shows distinctive effects on resorption geometry and on reversal phase activities which positively affect osteoblast recruitment and may therefore favor bone formation. © 2013 The Author(s).

Hamid O.,Angeles Clinic and Research Institute | Robert C.,Institute Gustave Roussy | Daud A.,University of California at San Francisco | Hodi F.S.,Dana-Farber Cancer Institute | And 22 more authors.
New England Journal of Medicine | Year: 2013

BACKGROUND: The programmed death 1 (PD-1) receptor is a negative regulator of T-cell effector mechanisms that limits immune responses against cancer. We tested the anti-PD-1 antibody lambrolizumab (previously known as MK-3475) in patients with advanced melanoma. METHODS: We administered lambrolizumab intravenously at a dose of 10 mg per kilogram of body weight every 2 or 3 weeks or 2 mg per kilogram every 3 weeks in patients with advanced melanoma, both those who had received prior treatment with the immune checkpoint inhibitor ipilimumab and those who had not. Tumor responses were assessed every 12 weeks. RESULTS: A total of 135 patients with advanced melanoma were treated. Common adverse events attributed to treatment were fatigue, rash, pruritus, and diarrhea; most of the adverse events were low grade. The confirmed response rate across all dose cohorts, evaluated by central radiologic review according to the Response Evaluation Criteria in Solid Tumors (RECIST), version 1.1, was 38% (95% confidence interval [CI], 25 to 44), with the highest confirmed response rate observed in the cohort that received 10 mg per kilogram every 2 weeks (52%; 95% CI, 38 to 66). The response rate did not differ significantly between patients who had received prior ipilimumab treatment and those who had not (confirmed response rate, 38% [95% CI, 23 to 55] and 37% [95% CI, 26 to 49], respectively). Responses were durable in the majority of patients (median follow-up, 11 months among patients who had a response); 81% of the patients who had a response (42 of 52) were still receiving treatment at the time of analysis in March 2013. The overall median progression-free survival among the 135 patients was longer than 7 months. CONCLUSIONS: In patients with advanced melanoma, including those who had had disease progression while they had been receiving ipilimumab, treatment with lambrolizumab resulted in a high rate of sustained tumor regression, with mainly grade 1 or 2 toxic effects. Copyright © 2013 Massachusetts Medical Society.

Girling P.R.,Durham University | Kiyoi T.,Merck And Co. | Whiting A.,Durham University
Organic and Biomolecular Chemistry | Year: 2011

A review into the aza-Diels-Alder reaction, mainly concentrating on literature examples that form piperidin-4-ones from the reaction of imines and electron rich dienes or enones, either through a Lewis acidic/Brønsted acid approach or through the use of an organocatalyst. This review questions whether the mechanism of the aza-Diels-Alder reaction is step wise as opposed to concerted when using oxygenated dienes. © 2011 The Royal Society of Chemistry.

Bauer D.C.,University of California at San Francisco | Schwartz A.,University of California at San Francisco | Palermo L.,University of California at San Francisco | Cauley J.,University of Pittsburgh | And 4 more authors.
JAMA Internal Medicine | Year: 2014

IMPORTANCE: Discontinuation of bisphosphonate therapy after 3 to 5 years is increasingly considered, but methods to monitor fracture risk after discontinuation have not been established. OBJECTIVE: To test methods of predicting fracture risk among women who have discontinued alendronate therapy after 4 to 5 years. DESIGN, SETTING, AND PARTICIPANTS: The prospective Fracture Intervention Trial Long-term Extension (FLEX) study randomized postmenopausal women aged 61 to 86 years previously treated with 4 to 5 years of alendronate therapy to 5 more years of alendronate or placebo from 1998 through 2003; the present analysis includes only the placebo group. Hip and spine dual-energy x-ray absorptiometry (DXA) were measured when placebo was begun (FLEX baseline) and after 1 to 3 years of follow-up. Two biochemical markers of bone turnover, urinary type 1 collagen cross-linked N-telopeptide (NTX) and serum bone-specific alkaline phosphatase (BAP), were measured at FLEX baseline and after 1 and 3 years. MAIN OUTCOMES AND MEASURES: Symptomatic spine and nonspine fractures occurring after the follow-up measurement of DXA or bone turnover. RESULTS: During 5 years of placebo, 94 of 437 women (22%) experienced 1 or more symptomatic fractures; 82 had fractures after 1 year. One-year changes in hip DXA, NTX, and BAP were not related to subsequent fracture risk, but older age and lower hip DXA at time of discontinuation were significantly related to increased fracture risk (lowest tertile of baseline femoral neck DXA vs other 2 tertiles relative hazard ratio, 2.17 [95%CI, 1.38-3.41]; total hip DXA relative hazard ratio, 1.87 [95%CI, 1.20-2.92]). CONCLUSIONS AND RELEVANCE: Among postmenopausalwomen who discontinue alendronate therapy after 4 to 5 years, age and hip BMD at discontinuation predict clinical fractures during the subsequent 5 years. Follow-up measurements of DXA 1 year after discontinuation and of BAP or NTX 1 to 2 years after discontinuation are not associated with fracture risk and cannot be recommended. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00398931 Copyright 2014 American Medical Association. All rights reserved.

Sotillo E.,Children's Hospital of Philadelphia | Laver T.,University of Pennsylvania | Mellert H.,University of Pennsylvania | Schelter J.M.,Rosetta Inpharmatics | And 7 more authors.
Oncogene | Year: 2011

Downregulation of microRNA-34a by Myc is known to be essential for tumorigenesis and improve tumor-cell survival. Conversely, upregulation of miR-34a by p53 is thought to enhance its acetylation and activity and contribute to the pro-apoptotic effects of this tumor suppressor. We sought to determine whether restoration of miR-34a levels in B-lymphoid cells with Myc overexpression would aid therapeutic apoptosis. Unexpectedly, delivery of miR-34a, which doesn't target p53 directly, severely compromised steady-state p53 levels. This effect was preceded and mediated by direct targeting of Myc, which sustained p53 protein levels via the Arf-Hdm2 pathway. As a result, in the presence of Myc, miR-34a inhibited p53-dependent bortezomib-induced apoptosis as efficiently as anti-p53 small interfering RNA. Conversely, inhibition of miR-34a using antisense RNA sensitized lymphoma cells to therapeutic apoptosis. Thus, in tumors with deregulated Myc expression, miR-34a confers drug resistance and could be considered a therapeutic target. © 2011 Macmillan Publishers Limited All rights reserved.

Engelke K.,Synarc Inc | Engelke K.,Friedrich - Alexander - University, Erlangen - Nuremberg | Fuerst T.,Synarc Inc | Dardzinski B.,Uniformed Services University of the Health Sciences | And 4 more authors.
Journal of Bone and Mineral Research | Year: 2015

Odanacatib, a selective cathepsin K inhibitor, increases areal bone mineral density (aBMD) at the spine and hip of postmenopausal women. To gain additional insight into the effects on trabecular and cortical bone, we analyzed quantitative computed tomography (QCT) data of postmenopausal women treated with odanacatib using Medical Image Analysis Framework (MIAF; Institute of Medical Physics, University of Erlangen, Erlangen, Germany). This international, randomized, double-blind, placebo-controlled, 2-year, phase 3 trial enrolled 214 postmenopausal women (mean age 64 years) with low aBMD. Subjects were randomized to odanacatib 50mg weekly (ODN) or placebo (PBO); all participants received calcium and vitamin D. Hip QCT scans at 24 months were available for 158 women (ODN: n=78 women; PBO: n=80 women). There were consistent and significant differential treatment effects (ODN-PBO) for total hip integral (5.4%), trabecular volumetric BMD (vBMD) (12.2%), and cortical vBMD (2.5%) at 24 months. There was no significant differential treatment effect on integral bone volume. Results for bone mineral content (BMC) closely matched those for vBMD for integral and trabecular compartments. However, with small but mostly significant differential increases in cortical volume (1.0% to 1.3%) and thickness (1.4% to 1.9%), the percentage cortical BMC increases were numerically larger than those of vBMD. With a total hip BMC differential treatment effect (ODN-PBO) of nearly 1000 mg, the proportions of BMC attributed to cortical gain were 45%, 44%, 52%, and 40% for the total, neck, trochanter, and intertrochanter subregions, respectively. In postmenopausal women treated for 2 years, odanacatib improved integral, trabecular, and cortical vBMD and BMC at all femur regions relative to placebo when assessed by MIAF. Cortical volume and thickness increased significantly in all regions except the femoral neck. The increase in cortical volume and BMC paralleled the increase in cortical vBMD, demonstrating a consistent effect of ODN on cortical bone. Approximately one-half of the absolute BMC gain occurred in cortical bone. © 2014 American Society for Bone and Mineral Research.

Thomas D.M.,Montclair State University | Schoeller D.A.,University of Wisconsin - Madison | Redman L.A.,Pennington Biomedical Research Center | Martin C.K.,Pennington Biomedical Research Center | And 2 more authors.
American Journal of Clinical Nutrition | Year: 2010

Background: Energy intake (EI) during weight loss is difficult and costly to measure accurately. Objective: The objective was to develop and validate a computational energy balance differential equation model to determine individual EI during weight loss. Design: An algorithm was developed to quantify EI during weight loss based on a validated one-dimensional model for weight change. By using data from a 24-wk calorie-restriction study, we tested the validity of the EI model against 2 criterion measures: 1) EI quantified through food provision from weeks 0-4 and 4-12 and 2) EI quantified through changes in body energy stores [measured with dual-energy X-ray absorptiometry (DXA)] and energy expenditure [measured with doubly labeled water (DLW)] from weeks 4-12 and 12-24. Results: Compared with food provision, the mean (6SD) model errors were 41 ± 118 kcal/d and -22 ± 230 kcal/d from weeks 0-4 and 4-12, respectively. Compared with EI measured with DXA and DLW, the model errors were -71 ± 272 kcal/d and -48 ± 226 kcal/d from weeks 4-12 and 12-24, respectively. In every comparison, the mean error was never significantly different from zero (P values > 0.10). Furthermore, Bland and Altman analysis indicated that error variance did not differ significantly over amounts of EI (P values > 0.26). Almost all individual participants' values were within CI limits. Conclusion: The validity of the newly developed EI model was supported by experimental observations and can be used to determine an individual participant's EI during weight loss. © 2010 American Society for Nutrition.

Shono Y.,Goethe University Frankfurt | Jantratid E.,Goethe University Frankfurt | Kesisoglou F.,Merck And Co. | Reppas C.,National and Kapodistrian University of Athens | Dressman J.B.,Goethe University Frankfurt
European Journal of Pharmaceutics and Biopharmaceutics | Year: 2010

This study coupled results from biorelevant dissolution tests with in silico simulation technology to forecast in vivo oral absorption of micronized and nanosized aprepitant formulations in the pre- and post-prandial states. In vitro dissolution tests of the nanosized aprepitant formulation and micronized drug were performed in biorelevant and compendial media. An in silico physiologically based pharmacokinetic (PBPK) model was developed based on STELLA® software using dissolution kinetics, standard gastrointestinal (GI) parameters and post-absorptive disposition parameters. GI parameters (gastric emptying rate and GI fluid volume) were varied according to the dosing conditions. Disposition parameters were estimated by fitting compartmental models to the in vivo oral PK data. Predictions of in vivo performance in each prandial state were evaluated using the AUC and Cmax generated from the simulated PK profiles. To predict oral absorption from the extremely fast dissolving nanosized aprepitant formulation, several variations on a previously published model were evaluated. Although models that assumed that the formulation behaved as an oral solution or that adjusted the dissolution kinetics according to the different numbers of particles per gram between micronized and nanosized aprepitant generated profiles similar to the observed in vivo data in the fed state, simulated profiles for the fasted state showed much faster absorption than that observed in the in vivo data. This appeared to result from the assumption of no absorption restrictions in those models. To better predict in vivo performance in both fasted and fed states, a model that adds permeability restrictions to absorption was applied. This model not only simulated the in vivo profiles for aprepitant well in both prandial states, but also predicted the dependency of the pharmacokinetics on the dose and the particle size of aprepitant. In conclusion, a model based on STELLA® software combined with dissolution results in biorelevant media successfully forecasts the in vivo performance of both nanosized and micronized formulations of aprepitant in the fed and fasted states. Although dissolution is the primary limitation to the rate of absorption for micronized aprepitant, some permeability restrictions are revealed for the nanosized formulation. The results also indicate that biorelevant dissolution media have strong advantages over compendial media in forecasting the in vivo behavior of aprepitant. © 2010 Elsevier B.V.

Fabricatore A.N.,University of Pennsylvania | Fabricatore A.N.,Nutrisystem | Wadden T.A.,University of Pennsylvania | Higginbotham A.J.,University of Pennsylvania | And 4 more authors.
International Journal of Obesity | Year: 2011

Objective:Obesity is related to increased risk of several health complications, including depression. Many studies have reported improvements in mood with weight loss, but results have been equivocal. The present meta-analysis examined changes in symptoms of depression that were reported in trials of weight loss interventions. Between-groups comparisons of different weight loss methods (for example, lifestyle modification, diet-alone and pharmacotherapy) were examined, as were within-group changes for each treatment type.Method:MEDLINE was searched for articles published between 1950 and January 2009. Several obesity-related terms were intersected with terms related to depression. Results were filtered to return only studies of human subjects, published in English. Of 5971 articles, 394 were randomized controlled trials. Articles were excluded if they did not report mean changes in weight or symptoms of depression, included children or persons with psychiatric disorders (other than depression), or provided insufficient data for analysis. Thirty-one studies (n7937) were included. Two authors independently extracted a description of each study treatment, sample characteristics, assessment methods and changes in weight and symptoms of depression. Treatments were categorized as lifestyle modification, non-dieting, dietary counseling, diet-alone, exercise-alone, pharmacotherapy, placebo or control interventions.Results:Random effects models found that lifestyle modification was superior to control and non-dieting interventions for reducing symptoms of depression, and marginally better than dietary counseling and exercise-alone programs. Exercise-alone programs were superior to controls. No differences were found for comparisons of pharmacologic agents and placebos. Within-group analyses found significant reductions in symptoms of depression for nearly all active interventions. A meta-regression found no relationship between changes in weight and changes in symptoms of depression in lifestyle modification interventions.Conclusions:On average, obese individuals in weight loss trials experienced reductions in symptoms of depression. Future studies should examine incidence and resolution of clinically significant depressive disorders with weight loss interventions. © 2011 Macmillan Publishers Limited All rights reserved.

Dobard C.,Centers for Disease Control and Prevention | Sharma S.,Centers for Disease Control and Prevention | Parikh U.M.,Centers for Disease Control and Prevention | West R.,Centers for Disease Control and Prevention | And 10 more authors.
Science Translational Medicine | Year: 2014

Coitally delivered microbicide gels containing antiretroviral drugs are important for HIV prevention. However, to date, microbicides have contained entry or reverse transcriptase inhibitors that block early steps in virus infection and thus need to be given as a preexposure dose that interferes with sexual practices and may limit compliance. Integrase inhibitors block late steps after virus infection and therefore are more suitable for post-coital dosing. We first determined the kinetics of strand transfer in vitro and confirmed that integration begins about 6 hours after infection. We then used a repeat-challenge macaque model to assess efficacy of vaginal gels containing integrase strand transfer inhibitors when applied before or after simian/human immunodeficiency virus (SHIV) challenge. We showed that gel containing the strand transfer inhibitor L-870812 protected two of three macaques when applied 30 min before SHIV challenge. We next evaluated the efficacy of 1% raltegravir gel and demonstrated its ability to protect macaques when applied 3 hours after SHIV exposure (five of six protected; P < 0.05, Fisher's exact test). Breakthrough infections showed no evidence of drug resistance in plasma or vaginal secretions despite continued gel dosing after infection. We documented rapid vaginal absorption reflecting a short pharmacological lag time and noted that vaginal, but not plasma, virus load was substantially reduced in the breakthrough infection after raltegravir gel treatment. We provide a proof of concept that topically applied integrase inhibitors protect against vaginal SHIV infection when administered shortly before or 3 hours after virus exposure.

Kuznetsova O.M.,Merck And Co. | Tymofyeyev Y.,Quantitative science
Statistics in Medicine | Year: 2014

In open-label studies, partial predictability of permuted block randomization provides potential for selection bias. To lessen the selection bias in two-arm studies with equal allocation, a number of allocation procedures that limit the imbalance in treatment totals at a pre-specified level but do not require the exact balance at the ends of the blocks were developed. In studies with unequal allocation, however, the task of designing a randomization procedure that sets a pre-specified limit on imbalance in group totals is not resolved. Existing allocation procedures either do not preserve the allocation ratio at every allocation or do not include all allocation sequences that comply with the pre-specified imbalance threshold. Kuznetsova and Tymofyeyev described the brick tunnel randomization for studies with unequal allocation that preserves the allocation ratio at every step and, in the two-arm case, includes all sequences that satisfy the smallest possible imbalance threshold. This article introduces wide brick tunnel randomization for studies with unequal allocation that allows all allocation sequences with imbalance not exceeding any pre-specified threshold while preserving the allocation ratio at every step. In open-label studies, allowing a larger imbalance in treatment totals lowers selection bias because of the predictability of treatment assignments. The applications of the technique in two-arm and multi-arm open-label studies with unequal allocation are described. © 2013 John Wiley & Sons, Ltd.

Phung O.J.,Western University of Health Sciences | Sobieraj D.M.,University of Connecticut | Engel S.S.,Merck And Co. | Rajpathak S.N.,Merck And Co.
Diabetes, Obesity and Metabolism | Year: 2014

Aims: Guidelines for type 2 diabetes recommend add-on agents when metformin alone fails to provide adequate glycaemic control. However, early combination therapy may benefit health outcomes. We conducted a systematic review and meta-analysis to investigate this question. Methods: We searched MEDLINE and Cochrane CENTRAL (up to July 2012) without language restrictions. We sought randomized controlled trials (RCTs) evaluating initial combination therapy with metformin versus metformin monotherapy in patients with untreated type 2 diabetes. Weighted mean differences (WMDs) for changes from baseline and relative risks (RRs) [with 95% confidence intervals (CIs)] were calculated using random-effects model. Results: In 15 RCTs (N=6693), the mean age range was 48.4-62.7years; mean baseline glycosylated haemoglobin (A1c) was 7.2-9.9% and mean diabetes duration was 1.6-4.1years, with median follow-up of 6months and with 13 comparisons for A1c change, 14 comparisons for A1c goal attainment of <7% and 13 comparisons for change in fasting plasma glucose (FPG). Drugs combined with metformin included thiazolidinediones (TZDs), insulin secretagogues, dipeptidyl peptidase-4 (DPP-4) inhibitors or sodium glucose transporterase (SGLT-2) inhibitors. Compared to metformin alone, combination therapy with metformin provided statistically significant reductions in A1c (WMD -0.43%, 95% CI -0.56, -0.30), increases in attainment of A1c goal of less than 7% (RR 1.40, 95% CI 1.33-1.48) and reductions in FPG (WMD -14.30mg/dl, 95% CI -16.09, -12.51). Conclusions: These results suggest a potential benefit of initial combination therapy on glycaemic outcomes in diabetes compared to metformin monotherapy across a wide range of baseline A1c levels. Further research should explore if early combination treatment may also affect longer term health outcomes in diabetes. © 2013 John Wiley & Sons Ltd.

Palmer L.,Truven Health Analytics | White R.R.,Merck And Co. | Johnson B.H.,Truven Health Analytics | Fowler R.,Truven Health Analytics | Acosta C.J.,Merck And Co.
Transplantation | Year: 2014

OBJECTIVES: To evaluate health-care utilization and costs attributable to herpes zoster (HZ) within a population of patients with solid organ transplant (SOT). METHODS: Using administrative claims data, a commercially/Medicare- insured population of patients with SOT between January 1, 1999, and January 1, 2007, and a Medicaid population between January 1, 1999, and January 1, 2006, were identified. Each patient group was screened to select patients with claims of SOT with an incident diagnosis of HZ and continuous enrollment for the 6 months prior and 3 months subsequent to the incident HZ. Controls were selected from group of SOT patients without claims of HZ using a propensity score matching process. Descriptive analyses were performed to quantify health-care utilization and costs attributable to HZ. Multivariate analyses were used to estimate HZ-Attributable costs adjusted by demographic and clinical variables. RESULTS: A total of 205 commercially/Medicare-insured matched pairs and 136 Medicaid matched pairs were selected. Mean age in the commercial/Medicare SOT-HZ population was 56.9 years, and that in the Medicaid population was 42.5 years. The majority of HZ patients were diagnosed within 2 years of evidence of SOT. The unadjusted differences in total HZ-Attributable health-care costs were $4762 and $6705 for commercial/Medicare-insured and Medicaid patients, respectively (P=0.176 and P=0.003, respectively) and were largely driven by hospitalization costs. Adjusted incremental costs in the SOT-HZ commercial/Medicare-insured patients were $5335 (P<0.001), and that in noncapitated Medicaid patients were $3711 (P<0.001). CONCLUSION: The occurrence of HZ in patients immunocompromised by SOT significantly increased health-care utilization and costs.Copyright © 2014 by Lippincott Williams & Wilkins.

Sun H.,University of California at San Francisco | Sun H.,Merck And Co. | Frassetto L.A.,University of California at San Francisco | Huang Y.,University of California at San Francisco | Benet L.Z.,University of California at San Francisco
Clinical Pharmacology and Therapeutics | Year: 2010

Nonrenal clearance of drugs can be significantly lower in patients with end-stage renal disease (ESRD) than in those with normal renal function. Using erythromycin (ER) as a probe compound, we investigated whether this decrease in nonrenal clearance is due to reduced hepatic clearance (CLH) and/or gut metabolism. We also examined the potential effects of the uremic toxins 3-carboxy-4-methyl-5-propyl-2-furan propanoic acid (CMPF) and indoxyl sulfate (Indox) on ER disposition. Route-randomized, two-way crossover pharmacokinetic studies of ER were conducted in 12 ESRD patients and 12 healthy controls after oral (250mg) and intravenous (125mg) dosing with ER. In patients with ESRD, CLH decreased 31% relative to baseline values (0.35±0.14l/h/kg vs. 0.51±0.13l/h/kg, P = 0.01), with no change in steady-state volume of distribution. With oral dosing, the bioavailability of ER increased 36% in patients with ESRD, and this increase was not related to changes in gut availability. As expected, plasma levels of CMPF and Indox were significantly higher in the patients than in the healthy controls. However, no correlation was observed between CLH of ER and the levels of uremic toxins. © 2010 American Society2010 American Society for Clinical Pharmacology and Therapeutics.

Breen S.M.,University of Iowa | Andric N.,University of Iowa | Ping T.,University of Iowa | Xie F.,University of California at San Francisco | And 3 more authors.
Molecular Endocrinology | Year: 2013

The LH receptor (LHR) activates several families of heterotrimeric G proteins, but only the activation of Gs and subsequent generation of cAMP are universally accepted as important mediators of LH actions. To examine the involvement of the Gq/11 family on the actions of LH, we crossed Cyp19Cre+ and Gαq f/f;Gα11 -/- mice to generate mice with a granulosa cell-specific deletion of Gαq in the context of a global deletion of Gα11. Granulosa cells from Gαq f/f;Gα11 -/-;Cre++ mice have barely detectable levels of Gαq/11, have a normal complement of LHR, and respond to LHR activation with a transient increase in cAMP accumulation, but they fail to respond with increased inositol phosphate accumulation, an index of the activation of Gαq/11. The LHR-provoked resumption of meiosis, cumulus expansion, and luteinization are normal. However, the Gαq f/f;Gα11 -/-; Cre++ mice display severe subfertility because many of the oocytes destined for ovulation become entrapped in preovulatory follicles or corpora lutea. Because follicular rupture is known to be dependent on the expression of the progesterone receptor (Pgr), we examined the LHR-induced expression of Pgr and 4 of its target genes (Adamts-1, Ctsl1, Edn2, and Prkg2). These actions of the LHR were impaired in the ovaries of the Gαq f/f;Gα11 -/-;Cre++ mice. We conclude that the defect in follicular rupture is secondary to the failure of the LHR to fully induce the expression of the Pgr. This is the first conclusive evidence for the physiological importance of the activation of Gq/11 by the LHR and for the involvement of Gαq/11 in ovulation. © 2013 by The Endocrine Society.

Girling P.R.,Durham University | Batsanov A.S.,Durham University | Shen H.C.,Merck And Co. | Whiting A.,Durham University
Chemical Communications | Year: 2012

Reaction of methoxyvinylmethylketone with different amines and aldehydes under Lewis-acid catalysed conditions results in a novel, formal, step-wise [1+2+1+2]-cycloaddition to give dihydropyridine products. © 2012 The Royal Society of Chemistry.

Ho T.W.,Merck And Co. | Ho T.W.,Astrazeneca | Connor K.M.,Merck And Co. | Zhang Y.,Merck And Co. | And 9 more authors.
Neurology | Year: 2014

Objective: To evaluate whether the calcitonin gene-related peptide (CGRP) receptor antagonist telcagepant might be effective for migraine prevention. Methods: In this randomized, double-blind, placebo-controlled, multicenter trial (ClinicalTrials.gov NCT00797667), patients experiencing 3-14 migraine days during a 4-week baseline were randomized to telcagepant 140 mg, telcagepant 280 mg, or placebo twice daily for 12 weeks. Efficacy was assessed by mean monthly headache days and migraine/probable migraine days (headache plus ≥1 associated symptom). Results: The trial was terminated following a recommendation from the Safety Monitoring Board due to hepatotoxicity concerns. At termination, the planned 660 patients had been randomized, 656 had been treated with ≥1 dose of study medication, and 14 had completed the trial. The mean treatment duration was 48-50 days. Thirteen patients, all in the telcagepant groups, had an alanine aminotransferase (ALT) elevation ≥33the upper limit of normal and 7 of these also had an aspartate aminotransferase elevation ≥33 the upper limit of normal. Two patients had very high symptomatic transaminase elevations that occurred within 2-6 weeks of treatment initiation and resolved after treatment discontinuation. The originally planned efficacy analysis over 12 weeks was not performed due to limited data at later time points, but there was evidence that telcagepant resulted in a larger reduction from baseline than placebo for mean monthly headache days (month 1: 140 mg=-2.9, 280 mg=-3.1, placebo=-1.7; p < 0.05) and migraine/probable migraine days (month 1: 140 mg=-2.7, 280 mg=-3.0, placebo=-1.6; p < 0.05). Conclusions: These data suggest a potential role for CGRP receptor antagonism in migraine prophylaxis. However, the observed aminotransferase elevations do not support the use of telcagepant for daily administration. Classification of evidence: This study provides Class II evidence that in patients with migraine, telcagepant taken daily reduces headache days by 1.4 days per month compared to placebo and causes 2.5% of patients to have elevations of serum ALT levels. © 2014 American Academy of Neurology.

Gray G.E.,University of Witwatersrand | Gray G.E.,South African Medical Research Council | Moodie Z.,Fred Hutchinson Cancer Research Center | Metch B.,Fred Hutchinson Cancer Research Center | And 15 more authors.
The Lancet Infectious Diseases | Year: 2014

Background: The HVTN 503/Phambili study, which assessed the efficacy of the Merck Ad5 gag/pol/nef subtype B HIV-1 preventive vaccine in South Africa, was stopped when futility criteria in the Step study (assessing the same vaccine in the Americas, Caribbean, and Australia) were met. Here we report long-term follow-up data. Methods: HVTN 503/Phambili was a double-blind, placebo-controlled, randomised trial that recruited HIV-1 uninfected, sexually active adults aged 18-35 years from five sites in South Africa. Eligible participants were randomly assigned (1:1) by computer-generated random numbers to either vaccine or placebo, stratified by site and sex. Cox proportional hazards models were used to estimate HIV-1 infection in the modified intention-to-treat cohort, all of whom were unmasked early in follow-up. The trial is registered with ClinicalTrials.gov, number NCT00413725 and the South African National Health Research Database, number DOH-27-0207-1539. Findings: Between Jan 24, 2007, and Sept 19, 2007, 801 participants (26·7%) of a planned 3000 were randomly assigned (400 to vaccine, 401 to placebo); 216 (27%) received only one injection, 529 (66%) received only two injections, and 56 (7%) received three injections. At a median follow-up of 42 months (IQR 31-42), 63 vaccine recipients (16%) had HIV-1 infection compared with 37 placebo recipients (9%; adjusted HR 1·70, 95% CI 1·13-2·55; p=0·01). Risk for HIV-1 infection did not differ according to the number of vaccinations received, sex, circumcision, or adenovirus type 5 (Ad5) serostatus. Differences in risk behaviour at baseline or during the study, or annualised dropout rate (7·7% [95% CI 6·2-9·5] for vaccine recipients vs 8·8% [7·1-10·7] for placebo recipients; p=0·40) are unlikely explanations for the increased rate of HIV-1 infections seen in vaccine recipients. Interpretation: The increased risk of HIV-1 acquisition in vaccine recipients, irrespective of number of doses received, warrants further investigation to understand the biological mechanism. We caution against further use of the Ad5 vector for HIV vaccines. Funding: National Institute of Allergy and Infectious Diseases, Merck, and South African Medical Research Council. © 2014 Elsevier Ltd.

Moriceau G.,University of California at Los Angeles | Hugo W.,University of California at Los Angeles | Hong A.,University of California at Los Angeles | Shi H.,University of California at Los Angeles | And 15 more authors.
Cancer Cell | Year: 2015

Combined BRAF- and MEK-targeted therapy improves upon BRAF inhibitor (BRAFi) therapy but is still beset by acquired resistance. We show that melanomas acquire resistance to combined BRAF and MEK inhibition by augmenting or combining mechanisms of single-agent BRAFi resistance. These double-drug resistance-associated genetic configurations significantly altered molecular interactions underlying MAPK pathway reactivation. V600EBRAF, expressed at supraphysiological levels because of V600EBRAF ultra-amplification, dimerized with and activated CRAF. In addition, MEK mutants enhanced interaction with overexpressed V600EBRAF via a regulatory interface at R662 of V600EBRAF. Importantly, melanoma cell lines selected for resistance to BRAFi+MEKi, but not those to BRAFi alone, displayed robust drug addiction, providing a potentially exploitable therapeutic opportunity. © 2015 Elsevier Inc.

Dankovic D.A.,Centers for Disease Control and Prevention | Naumann B.D.,Merck And Co. | Maier A.,University of Cincinnati | Dourson M.L.,University of Cincinnati | Levy L.S.,Cranfield University
Journal of Occupational and Environmental Hygiene | Year: 2015

The uncertainty factor concept is integrated into health risk assessments for all aspects of public health practice, including by most organizations that derive occupational exposure limits. The use of uncertainty factors is predicated on the assumption that a sufficient reduction in exposure from those at the boundary for the onset of adverse effects will yield a safe exposure level for at least the great majority of the exposed population, including vulnerable subgroups. There are differences in the application of the uncertainty factor approach among groups that conduct occupational assessments; however, there are common areas of uncertainty which are considered by all or nearly all occupational exposure limit-setting organizations. Five key uncertainties that are often examined include interspecies variability in response when extrapolating from animal studies to humans, response variability in humans, uncertainty in estimating a no-effect level from a dose where effects were observed, extrapolation from shorter duration studies to a full life-time exposure, and other insufficiencies in the overall health effects database indicating that the most sensitive adverse effect may not have been evaluated. In addition, a modifying factor is used by some organizations to account for other remaining uncertainties-typically related to exposure scenarios or accounting for the interplay among the five areas noted above. Consideration of uncertainties in occupational exposure limit derivation is a systematic process whereby the factors applied are not arbitrary, although they are mathematically imprecise. As the scientific basis for uncertainty factor application has improved, default uncertainty factors are now used only in the absence of chemical-specific data, and the trend is to replace them with chemical-specific adjustment factors whenever possible. The increased application of scientific data in the development of uncertainty factors for individual chemicals also has the benefit of increasing the transparency of occupational exposure limit derivation. Improved characterization of the scientific basis for uncertainty factors has led to increasing rigor and transparency in their application as part of the overall occupational exposure limit derivation process. © 2015 D. A. Dankovic, B. D. Naumann, A. Maier, M. L. Dourson and L. S. Levy.

Feldstein A.C.,Kaiser Permanente | Black D.,University of California at San Francisco | Perrin N.,Kaiser Permanente | Rosales A.G.,Kaiser Permanente | And 7 more authors.
Journal of Bone and Mineral Research | Year: 2012

The case definition, community incidence, and characteristics of atypical femoral shaft fractures (FSFs) are poorly understood. This retrospective study utilized electronic medical records and radiograph review among women ≥50 years of age and men ≥65 years of age from January 1996 to June 2009 at Kaiser Permanente Northwest to describe the incidence rates and characteristics of subgroups of femur fractures. Fractures were categorized based on the American Society for Bone and Mineral Research (ASBMR) as atypical fracture major features (AFMs) (low force, shaft location, transverse or short oblique, noncomminuted) and AFMs with additional minor radiograph features (AFMms) (beaking, cortical thickening, or stress fracture). There were 5034 fractures in the study. The incidence rates of FSFs (without atypical features) and AFMs appeared flat (cumulative incidence: 18.2 per 100,000 person-years, 95% CI=16.0-20.7; 5.9 per 100,000 person-years, 95% CI=4.6-7.4; respectively) with 1,271,575 person-years observed. The proportion of AFMs that were AFMms increased over time. Thirty percent of AFMs had any dispensing of a bisphosphonate prior to the fracture, compared to 15.8% of the non-atypical FSFs. Years of oral glucocorticosteroid dispensing appeared highest in AFM and AFMm fractures. Those with AFMs only were older and had a lower frequency of bisphosphonate dispensing compared to those with AFMms. We conclude that rates of FSFs, with and without atypia, were low and stable over 13.5 years. Patients with only AFMs appear to be different from those with AFMms; it may be that only the latter group is atypical. There appear to be multiple associated risk factors for AFMm fractures. © 2012 American Society for Bone and Mineral Research.

Agency: European Commission | Branch: FP7 | Program: MC-ITN | Phase: FP7-PEOPLE-2011-ITN | Award Amount: 3.87M | Year: 2012

The Earth receives the energy in 1 hour required for all human needs in a year. Harvesting solar energy will reduce harmful CO2 emissions and resolve the forthcoming energy deficit that other sources alone cannot make up. The market for stable, cheap, roll-to-roll mass-produced organic solar cells (OSCs) is estimated at 1 billion Euros by 2016. The ITN ESTABLIS will produce a team of 11 ESRs and 4 ERs to harness this pivotal point in Europes development based on a reliable, economically powerful and clean resource. ESTABLIS will be an interdisciplinary and inter-sectorial research and training network. ESRs and ERs that result from ESTABLIS will excel. They will possess a broad skill-set across a range of disciplines that are of absolute necessity to develop the industrial and academic infra-structure in OSCs. Researchers will receive training in the primary areas of synthetic organic chemistry through complementary aspects of polymer science to complete industrial scale photovoltaic device manufacture. To improve the roll-to-roll engineering and stability of opto-electronically active thin-films will require new polymers, surface treatments, rheological appraisals of polymer processing, and ageing studies. A parallel approach will develop the necessary improvements in electronic and opto-electronic properties by clarifying correlations between charge transfer, photochemistry and stability. This project will be run by meticulously interacting groups to increase the stability of strong, flexible, low-cost OSCs to 10 years so that they can be sold on a mass-market basis. ESTABLIS is an exceptionally complementary consortium of field-leading University groups and the worlds chief Industrial companies, namely, the worlds foremost producers of OSCs, Konarka, of conducting polymers, Heraeus, and of semiconducting polymers, Merck. These key European companies are at the heart to ensure that training and technological developments will be industrially operable.

Ma S.,Genentech | Chowdhury S.K.,Merck And Co.
Bioanalysis | Year: 2012

Biotransformation of chemically stable compounds to reactive metabolites that can bind covalently to macromolecules (such as proteins and DNA) is considered an undesirable property of drug candidates. Due to the possible link, which has not yet been conclusively demonstrated, between reactive metabolites and adverse drug reactions, screening for metabolic activation of lead compounds through in vitro chemical trapping experiments has become an integral part of the drug discovery process in many laboratories. In this review, we provide an overview of the recent advances in the application of high-resolution MS. These advances facilitated the development of accurate-mass-based data mining tools for high-throughput screening of reactive drug metabolites in drug discovery. © 2012 Future Science Ltd.

Koliwad S.K.,Gladstone | Koliwad S.K.,University of California at San Francisco | Streeper R.S.,Gladstone | Monetti M.,Gladstone | And 8 more authors.
Journal of Clinical Investigation | Year: 2010

Diet-induced obesity (DIO) leads to inflammatory activation of macrophages in white adipose tissue (WAT) and subsequently to insulin resistance. PPARγ agonists are antidiabetic agents known to suppress inflammatory macrophage activation and to induce expression of the triacylglycerol (TG) synthesis enzyme acyl CoA: diacylglycerol acyltransferase 1 (DGAT1) in WAT and in adipocytes. Here, we investigated in mice the relationship between macrophage lipid storage capacity and DIO-associated inflammatory macrophage activation. Mice overexpressing DGAT1 in both macrophages and adipocytes (referred to herein as aP2-Dgat1 mice) were more prone to DIO but were protected against inflammatory macrophage activation, macrophage accumulation in WAT, systemic inflammation, and insulin resistance. To assess the contribution of macrophage DGAT1 expression to this phenotype, we transplanted wild-type mice with aP2-Dgat1 BM. These mice developed DIO similar to that of control mice but retained the protection from WAT inflammation and insulin resistance seen in aP2-Dgat1 mice. In isolated macrophages, Dgat1 mRNA levels correlated directly with TG storage capacity and inversely with inflammatory activation by saturated fatty acids (FAs). Moreover, PPARγ agonists increased macrophage Dgat1 mRNA levels, and the protective effects of these agonists against FA-induced inflammatory macrophage activation were absent in macrophages isolated from Dgat1-null mice. Thus, increasing DGAT1 expression in murine macrophages increases their capacity for TG storage, protects against FA-induced inflammatory activation, and is sufficient to reduce the inflammatory and metabolic consequences of DIO.

Chen C.,Merck And Co. | Beckman R.A.,University of California at San Francisco | Beckman R.A.,Daiichi Sankyo
Clinical Cancer Research | Year: 2014

Phase II proof-of-concept (POC) trials play a key role in oncology drug development, determining which therapeutic hypotheses will undergo definitive phase III testing according to predefined Go-No Go (GNG) criteria. The number of possible POC hypotheses likely far exceeds available public or private resources. We propose a design strategy for maximizing return on socioeconomic investment in phase II trials that obtains the greatest knowledge with the minimum patient exposure. We compare efficiency using the benefit-cost ratio, defined to be the risk-adjusted number of truly active drugs correctly identified for phase III development divided by the risk-adjusted total sample size in phase II and III development, for different POC trial sizes, powering schemes, and associated GNG criteria. It is most cost-effective to conduct small POC trials and set the corresponding GNG bars high, so that more POC trials can be conducted under socioeconomic constraints. If δ is the minimum treatment effect size of clinical interest in phase II, the study design with the highest benefit-cost ratio has approximately 5% type I error rate and approximately 20% type II error rate (80% power) for detecting an effect size of approximately 1.5δ. A Go decision to phase III is made when the observed effect size is close to δ. With the phenomenal expansion of our knowledge in molecular biology leading to an unprecedented number of new oncology drug targets, conducting more small POC trials and setting high GNG bars maximize the return on socioeconomic investment in phase II POC trials. ©2014 AACR.

Domar A.,Beth Israel Deaconess Medical Center | Gordon K.,Merck And Co. | Garcia-Velasco J.,Rey Juan Carlos University | La Marca A.,Section of Obstetrics and Gynecology | And 2 more authors.
Human Reproduction | Year: 2012

Background Infertility can significantly impact womens lives and personal relationships. Despite the negative impact of infertility, a significant number of women who are struggling to conceive do not consult a physician. This cross-sectional survey was conducted to determine the emotional impact of infertility on women to identify which aspects of fertility treatment contribute to the psychological stress experienced by so many patients and to identify barriers to seeking treatment.Methods Women (n = 445; 1844 years) who had received fertility treatment within the past 2 years or were having trouble conceiving but had not received treatment, completed a 15-min survey online. Results Participants were from France (n = 108), Germany (n = 111), Italy (n = 112) and Spain (n = 114). Responses indicated that infertility causes a range of emotions and can strain relationships. Women who had received treatment were more likely to feel hopeful (26 versus 21) and closer to their partner than women not in treatment (33 versus 19, P < 0.05). Most women delayed starting treatment because of a desire to conceive naturally, and on the advice of physicians. Women aged <35 years took longer to seek help with their fertility issues. Injection-related anxiety was the second greatest barrier to treatment. Conclusion s This study has provided insight into the physical and psychological challenges of infertility treatments and permitted a better understanding of the factors that impact patient lives. A treatment protocol with minimal injections and provision of additional information may lessen the emotional impact and challenges of infertility and contribute to patient satisfaction with fertility treatment protocols. © 2012 The Author.

MUMBAI, India and PRINCETON, N.J., March 1, 2017 /PRNewswire/ -- Sun Pharmaceutical Industries Ltd (Reuters: SUN.BO, Bloomberg: SUNP IN, NSE: SUNPHARMA, BSE: 524715, "Sun Pharma" and includes its subsidiaries or associate companies) today announced that several new analyses from Phase-1...

Gordon S.C.,Ford Motor Company | Zuckerman E.,Carmel Medical Center | Lawitz E.,University of Texas Health Science Center at San Antonio | Calleja J.L.,Autonomous University of Madrid | And 9 more authors.
Journal of Hepatology | Year: 2015

Background & Aims The Phase-2 C-SALVAGE study evaluated an investigational interferon-free combination of grazoprevir (a NS3/4A protease inhibitor) and elbasvir (a NS5A inhibitor) with ribavirin for patients with chronic HCV genotype-1 infection who had failed licensed DAA-containing therapy. Methods C-SALVAGE was an open-label study of grazoprevir 100 mg and elbasvir 50 mg QD with weight-based ribavirin BID for 12 weeks in cirrhotic and non-cirrhotic patients with chronic HCV genotype-1 infection who had not attained SVR after ≥4 weeks of peginterferon and ribavirin plus either boceprevir, telaprevir, or simeprevir. Exclusion criteria included decompensated liver disease, hepatocellular carcinoma, and HIV or HBV co-infection. The primary efficacy outcome was SVR12 defined as a HCV RNA level below the assay limit of quantification 12 weeks after the end of treatment. Results Of the 79 patients treated with ≥1 dose of study drug, 66 (84%) patients had a history of virologic failure on a regimen containing a NS3/4A protease inhibitor; 12 of the other 13 patients discontinued prior treatment because of adverse experiences. At entry, 34 (43.6%) of 78 evaluable patients harbored NS3 RAVs. SVR12 rates were 76/79 (96.2%) overall, including 28/30 (93.3%) patients with genotype 1a infection, 63/66 (95.5%) patients with prior virologic failure, 43/43 (100%) patients without baseline RAVs, 31/34 (91.2%) patients with baseline NS3 RAVs, 6/8 (75.0%) patients with baseline NS5A RAVs, 4/6 (66.7%) patients with both baseline NS3 and RAVs, and 32/34 (94.1%) cirrhotic patients. None of the five reported serious adverse events were considered drug-related. Conclusions Grazoprevir and elbasvir plus ribavirin for 12 weeks provides a promising new treatment option for patients after failure of triple therapy containing an earlier-generation protease inhibitor. © 2015 European Association for the Study of the Liver.

Sulkowski M.,Johns Hopkins University | Hezode C.,French Institute of Health and Medical Research | Gerstoft J.,Rigshospitalet | Vierling J.M.,Baylor College of Medicine | And 18 more authors.
The Lancet | Year: 2015

Background Both hepatitis C virus (HCV) mono-infected and HIV/HCV co-infected patients are in need of safe, effective, all-oral HCV regimens. In a phase 2 study we aimed to assess the efficacy and safety of grazoprevir (MK-5172; HCV NS3/4A protease inhibitor) and two doses of elbasvir (MK-8742; HCV NS5A inhibitor) in patients with HCV mono-infection and HIV/HCV co-infection. Methods The C-WORTHY study is a phase 2, multicentre, randomised controlled trial of grazoprevir plus elbasvir with or without ribavirin in patients with HCV; here, we report findings for previously untreated (genotype 1) patients without cirrhosis who were HCV mono-infected or HIV/HCV co-infected. Eligible patients were previously untreated adults aged 18 years or older with chronic HCV genoype 1 infection and HCV RNA at least 10 000 IU/mL in peripheral blood without evidence of cirrhosis, hepatocellular carcinoma, or decompensated liver disease. In part A of the study we randomly assigned HCV-mono-infected patients to receive 12 weeks of grazoprevir (100 mg) plus elbasvir (20 mg or 50 mg) with or without ribavirin (arms A1-3); in part B we assigned HCV-mono-infected patients to 8 or 12 weeks of grazoprevir (100 mg) plus elbasvir (50 mg) with or without ribavirin (arms B1-3) and HIV/HCV co-infected patients to 12 weeks of therapy with or without ribavirin. The primary endpoint was the proportion of patients achieving HCV RNA less than 25 IU/mL 12 weeks after end of treatment (SVR12). Randomisation was by presence or absence of ribavirin, 8 or 12 weeks of treatment, and dosage of elbasvir. Patients were stratified by gentoype 1a versus 1b. The patients, investigators, and study site personnel were masked to treatment group assignements but the funder was not. Analysis was by intention to treat. This trial is registered at ClinicalTrials.gov, number NCT01717326. Findings 218 patients with HCV mono-infection (n=159) and HIV/HCV co-infection (n=59) were enrolled. SVR12 for patients treated for 12 weeks with or without ribavirin ranged from 93-98% in mono-infected and 87-97% in co-infected patients. SVR12 rates in mono-infected and co-infected patients treated for 12 weeks without ribavirin were 98% (95% CI 88-100; 43/44) and 87% (95% CI 69-96; 26/30), respectively, and with ribavirin were 93% (95% CI 85-97; 79/85) and 97% (95% CI 82-100; 28/29), respectively. Among mono-infected patients with genotype 1a infection treated for 8 weeks, SVR12 was 80% (95% CI 61-92; 24/30). Five of six patients who discontinued early for reasons other than virological failure had HCV RNA less than 25 IU/mL at their last study visit. Virological failure among patients treated for 12 weeks occurred in seven patients (7/188, 4%) and was associated with emergence of resistance-associated variants to one or both drugs. The safety profile of grazoprevir plus elbasvir with or without ribavirin was similar in mono-infected and co-infected patients. No patient discontinued due to an adverse event or laboratory abnormality. The most common adverse events were fatigue (51 patients, 23%), headache (44, 20%), nausea (32, 15%), and diarrhoea (21, 10%). Interpretation Once-daily grazoprevir plus elbasvir with or without ribavirin for 12 weeks in previously untreated HCV-mono-infected and HIV/HCV-co-infected patients without cirrhosis achieved SVR12 rates of 87-98%. These results support the ongoing phase 3 development of grazoprevir plus elbasvir. Funding Merck & Co, Inc. © 2015 Elsevier Ltd.

Jia T.,University of Pennsylvania | Bellomo A.,University of Pennsylvania | Baina K.E.,University of Pennsylvania | Dreher S.D.,Merck And Co. | Walsh P.J.,University of Pennsylvania
Journal of the American Chemical Society | Year: 2013

The palladium-catalyzed α-arylation of unactivated sulfoxides has been developed. The weakly acidic α-protons of sulfoxides are reversibly deprotonated by LiOtBu, and a palladium phosphine complex facilitates the arylation. A variety of aryl methyl sulfoxides were coupled with aryl bromides. More challenging coupling partners, such as alkyl methyl sulfoxides (including dimethyl sulfoxide) and aryl chlorides proved to be suitable under the optimized conditions. This method was utilized to synthesize bioactive benzyl sulfoxide intermediates. © 2013 American Chemical Society.

Aarts M.,Royal Marsden Hospital | Sharpe R.,Royal Marsden Hospital | Garcia-Murillas I.,Royal Marsden Hospital | Gevensleben H.,Royal Marsden Hospital | And 5 more authors.
Cancer Discovery | Year: 2012

Inhibition of the protein kinase WEE1 synergizes with chemotherapy in preclinical models and WEE1 inhibitors are being explored as potential cancer therapies. Here, we investigate the mechanism that underlies this synergy. We show that WEE1 inhibition forces S-phase-arrested cells directly into mitosis without completing DNA synthesis, resulting in highly abnormal mitoses characterized by dispersed chromosomes and disorganized bipolar spindles, ultimately resulting in mitotic exit with gross micronuclei formation and apoptosis. This mechanism of cell death is shared by CHK1 inhibitors, and combined WEE1 and CHK1 inhibition forces mitotic entry from S-phase in the absence of chemotherapy. We show that p53/p21 inactivation combined with high expression of mitotic cyclins and EZH2 predispose to mitotic entry during S-phase with cells reliant on WEE1 to prevent premature cyclin-dependent kinase (CDK)1 activation. These features are characteristic of aggressive breast, and other, cancers for which WEE1 inhibitor combinations represent a promising targeted therapy. SIGNIFICANCE: Here, we describe a novel mechanism of inducing cancer cell death by WEE1 inhibition, forcing mitotic entry directly from S-phase. This mechanism represents a potential therapeutic approach for aggressive breast cancers, and in particular triple-negative and basal-like breast cancers, as WEE1 inhibition specifically targets the features inherent in these cancers: frequent TP53 mutation and high expression of mitotic cyclins and the polycomb protein EZH2. © 2012 American Association for Cancer Research.

Metz A.E.,University of Pennsylvania | Berritt S.,University of Pennsylvania | Dreher S.D.,Merck And Co. | Kozlowski M.C.,University of Pennsylvania
Organic Letters | Year: 2012

Palladium-catalyzed cross-coupling conditions were developed that efficiently afford 2-aryl-2-nitroacetates from aryl bromides and the very acidic nitroacetates. © 2012 American Chemical Society.

Mazur M.T.,Merck And Co. | Mazur M.T.,Imclone Systems | Cardasis H.L.,Merck And Co.
Methods in Molecular Biology | Year: 2013

The fi eld of quantitative, label-free proteomics has evolved signi fi cantly over time, with most experiments performed "bottom-up" using proteolyzed protein mixtures. In these experiments, statistically signi fi cant peptide abundance differences between two or more experimental conditions are determined, and their corresponding proteins later identi fi ed. Recently, the rationale for extending this experimental design to mixtures of intact proteins has become clear, as analysis at the protein level allows for the independent detection of each protein form present, including those modi fi ed posttranslationally. This provides a level of speci fi city lost in bottom-up experiments. As such, the application of label-free top-down differential mass spectrometry has provided a means for understanding the subtle protein changes that de fi ne a particular phenotype. Described here is an approach for the top-down label-free quantitative analysis of the proteins which constitute human high-density lipoprotein particles. The methodology is conceptually very straightforward; however, it does require a level of rigor and consistency typically not addressed by more conventional proteomics experiments. © 2013 Springer Science+Business Media New York.

Cicchini M.,Rutgers University | Cicchini M.,University of Pennsylvania | Karantza V.,Rutgers University | Karantza V.,Merck And Co. | Xia B.,Rutgers University
Clinical Cancer Research | Year: 2015

Autophagy is an intracellular self-digestion mechanism, by which cellular components are sorted into double-membrane autophagosomes and delivered to lysosomes for degradation. Cells use autophagy to dispose of wastes and eliminate hazards, while recycling nutrients and tuning metabolism in the process. Through these functions, autophagy promotes cell fitness, genome integrity, tissue homeostasis, and cell survival and growth under stress. Both autophagy upregulation and downregulation have been found in human cancers, suggesting a complex role in tumor development. Accumulating results from autophagy-deficient mice and mouse models of human cancers have demonstrated that autophagy generally suppresses tumor initiation, but promotes tumor progression, in a manner that is dependent on timing and context and modified by specific tumorigenic events. Given the role of autophagy in facilitating tumor growth, autophagy inhibition has gained wide attention as a potential anticancer therapy. Here, we summarize relevant genetic, preclinical, and clinical studies and discuss the multifaceted role of autophagy in cancer, as well as the prospects of autophagy inhibition for cancer therapy. © 2014 AACR.

Zhang J.,University of Pennsylvania | Bellomo A.,University of Pennsylvania | Trongsiriwat N.,University of Pennsylvania | Jia T.,University of Pennsylvania | And 7 more authors.
Journal of the American Chemical Society | Year: 2014

Although the past 15 years have witnessed the development of sterically bulky and electron-rich alkylphosphine ligands for palladium-catalyzed cross-couplings with aryl chlorides, examples of palladium catalysts based on either triarylphosphine or bidentate phosphine ligands for efficient room temperature cross-coupling reactions with unactivated aryl chlorides are rare. Herein we report a palladium catalyst based on NiXantphos, a deprotonatable chelating aryldiphosphine ligand, to oxidatively add unactivated aryl chlorides at room temperature. Surprisingly, comparison of an extensive array of ligands revealed that under the basic reaction conditions the resultant heterobimetallic Pd-NiXantphos catalyst system outperformed all the other mono- and bidentate ligands in a deprotonative cross-coupling process (DCCP) with aryl chlorides. The DCCP with aryl chlorides affords a variety of triarylmethane products, a class of compounds with various applications and interesting biological activity. Additionally, the DCCP exhibits remarkable chemoselectivity in the presence of aryl chloride substrates bearing heteroaryl groups and sensitive functional groups that are known to undergo 1,2-addition, aldol reaction, and O-, N-, enolate-α-, and C(sp2)-H arylations. The advantages and importance of the Pd-NiXantphos catalyst system outlined herein make it a valuable contribution for applications in Pd-catalyzed arylation reactions with aryl chlorides. © 2014 American Chemical Society.

Cai B.,Merck And Co. | Cai B.,University of Pennsylvania | Small D.S.,University of Pennsylvania | Have T.R.T.,University of Pennsylvania
Statistics in Medicine | Year: 2011

We present closed-form expressions of asymptotic bias for the causal odds ratio from two estimation approaches of instrumental variable logistic regression: (i) the two-stage predictor substitution (2SPS) method and (ii) the two-stage residual inclusion (2SRI) approach. Under the 2SPS approach, the first stage model yields the predicted value of treatment as a function of an instrument and covariates, and in the second stage model for the outcome, this predicted value replaces the observed value of treatment as a covariate. Under the 2SRI approach, the first stage is the same, but the residual term of the first stage regression is included in the second stage regression, retaining the observed treatment as a covariate. Our bias assessment is for a different context from that of Terza (J. Health Econ. 2008; 27(3):531-543), who focused on the causal odds ratio conditional on the unmeasured confounder, whereas we focus on the causal odds ratio among compliers under the principal stratification framework. Our closed-form bias results show that the 2SPS logistic regression generates asymptotically biased estimates of this causal odds ratio when there is no unmeasured confounding and that this bias increases with increasing unmeasured confounding. The 2SRI logistic regression is asymptotically unbiased when there is no unmeasured confounding, but when there is unmeasured confounding, there is bias and it increases with increasing unmeasured confounding. The closed-form bias results provide guidance for using these IV logistic regression methods. Our simulation results are consistent with our closed-form analytic results under different combinations of parameter settings. © 2011 John Wiley & Sons, Ltd.

Herlogsson L.,Linköping University | Crispin X.,Linköping University | Tierney S.,Merck And Co. | Berggren M.,Linköping University
Advanced Materials | Year: 2011

Organic complementary inverters and ring oscillators based on polyelectrolyte-gated thin-film transistors are demonstrated. Detrimental electrochemical doping is suppressed by using polyanionic and polycationic gate insulators in the p- and n-channel transistors, respectively. The circuits operate at supply voltages between 0.2 V and 1.5 V, have a static power consumption of less than 2.5 nW per logic gate and show propagation delays down to 0.26 ms per stage. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

McGrew G.I.,University of Pennsylvania | Stanciu C.,University of Pennsylvania | Zhang J.,University of Pennsylvania | Carroll P.J.,University of Pennsylvania | And 2 more authors.
Angewandte Chemie - International Edition | Year: 2012

Ligand in a haystack: The first catalytic asymmetric cross-coupling of benzyllithiums α to tertiary amines using [Cr(CO)3] activation of benzylic C sp 3-H bonds is described. The stabilized organolithium undergoes Pd-catalyzed coupling with aryl triflates (ArOTf) by a novel dynamic kinetic resolution to yield enantioenriched diarylmethylamines. The chiral ligand for this reaction was identified using high-throughput experimentation with 192 ligands. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Bender K.J.,Oregon Health And Science University | Uebele V.N.,Merck And Co. | Renger J.J.,Merck And Co. | Trussell L.O.,Oregon Health And Science University
Journal of Physiology | Year: 2012

Spontaneously active neurons typically fire either in a regular pattern or in bursts. While much is known about the subcellular location and biophysical properties of conductances that underlie regular spontaneous activity, less is known about those that underlie bursts. Here, we show that T-type Ca 2+ channels localized to the site of action potential initiation in the axon initial segment play a pivotal role in spontaneous burst generation. In auditory brainstem interneurons, axon initial segment Ca 2+ influx is selectively downregulated by dopaminergic signalling. This regulation has marked effects on spontaneous activity, converting the predominant mode of spontaneous activity from bursts to regular spiking. Thus, the axon initial segment is a key site, and dopamine a key regulator, of spontaneous bursting activity. © 2012 The Authors. The Journal of Physiology © 2012 The Physiological Society.

Russell M.R.,Children's Hospital of Philadelphia | Levin K.,Children's Hospital of Philadelphia | Rader J.,Children's Hospital of Philadelphia | Belcastro L.,Children's Hospital of Philadelphia | And 9 more authors.
Cancer Research | Year: 2013

Neuroblastoma is uniquely sensitive to single-agent inhibition of the DNA damage checkpoint kinase Chk1, leading us to examine downstream effectors of this pathway and identify mitotic regulator Wee1 as an additional therapeutic target in this disease. Wee1 was overexpressed in both neuroblastoma cell lines and high-risk patient tumors. Genetic or pharmacologic abrogation of Wee1 signaling results in marked cytotoxicity in 10 of 11 neuroblastoma cell lines with a median IC50 of 300 nmol/L for the Wee1-selective small-molecule inhibitor MK- 1775. Murine tumor lines derived from mice that were either heterozygous or homozygous for MycN were particularly sensitive to single-agent inhibition of Wee1 (IC50s of 160 and 62 nmol/L, respectively). Simultaneous pharmacologic inhibition of Chk1 and Wee1 acted in a synergistic fashion to further impede neuroblastoma cell growth in vitro, in a manner greater than the individual inhibitors either alone or combined with chemotherapy. Combination Chk1 and Wee1 inhibition also revealed in vivo efficacy in neuroblastoma xenografts. Taken together, our results show that neuroblastoma cells depend on Wee1 activity for growth and that inhibition of this kinase may serve as a therapeutic for patients with neuroblastoma. © 2012 AACR.

Lido H.H.,Gothenburg University | Marston H.,Merck And Co. | Ericson M.,Gothenburg University | Soderpalm B.,Gothenburg University | Soderpalm B.,Sahlgrenska University Hospital
Addiction Biology | Year: 2012

Extracellular glycine modulates accumbal dopamine levels as well as ethanol-induced dopamine overflow. Glycine availability is also crucial for regulating alcohol consumption and the glycine transporter 1 (GlyT-1) inhibitor Org25935 robustly decreases alcohol intake in rats. To explore whether the alcohol-intake reducing effect of Org25935 is substance bound, we examined the effect of a different selective GlyT-1 inhibitor, Org24598, on ethanol consumption in rats and compared the effect with that of acamprosate, a drug currently in clinical use. We studied the effects of daily Org24598 and acamprosate injections on male Wistar rats with a ethanol preference in a limited access two bottle free-choice model for 12 days, followed by alcohol deprivation for 14 days before a second test period of 10 days. Finally, rats underwent in vivo microdialysis where dopamine, glycine, taurine and β-alanine in n. accumbens were measured. Org24598 profoundly reduced ethanol intake and the effect remained throughout both treatment periods. Acamprosate promptly reduced ethanol intake, but on the third day tolerance developed to this effect and acamprosate failed to influence alcohol consumption during the second test period. Neither Org24598 nor acamprosate reduced water intake. Following the drinking study, the Org24598 group displayed higher basal accumbal dopamine levels compared with acamprosate and vehicle groups. Both Org24598 and acamprosate reduced the ethanol-induced dopamine response in n. accumbens. The study demonstrates a robust anti-alcohol intake effect of the GlyT-1 inhibitor Org24598, supporting the new concept that GlyT-1 inhibition reduces ethanol consumption. GlyT-1 inhibition may represent a new treatment principle for alcoholism that is superior to acamprosate. © 2011 The Authors, Addiction Biology © 2011 Society for the Study of Addiction.

Qiao Y.,Hospital for Special Surgery | Giannopoulou E.G.,New York Medical College | Chan C.H.,Hospital for Special Surgery | Park S.-H.,Hospital for Special Surgery | And 8 more authors.
Immunity | Year: 2013

Synergistic activation of inflammatory cytokine genes by interferon-γ (IFN-γ) and Toll-like receptor (TLR) signaling is important for innate immunity and inflammatory disease pathogenesis. Enhancement of TLR signaling, a previously proposed mechanism, is insufficient to explain strong synergistic activation of cytokine production in human macrophages. Rather, we found that IFN-γ induced sustained occupancy of transcription factors STAT1, IRF-1, and associated histone acetylation at promoters and enhancers at the TNF, IL6, and IL12B loci. This priming of chromatin did not activate transcription but greatly increased and prolonged recruitment of TLR4-induced transcription factors and RNA polymerase II to gene promoters and enhancers. Priming sensitized cytokine transcription to suppression by Jak inhibitors. Genome-wide analysis revealed pervasive priming of regulatory elements by IFN-γ and linked coordinate priming of promoters and enhancers with synergistic induction of transcription. Our results provide a synergy mechanism whereby IFN-γ creates a primed chromatin environment to augment TLR-induced gene transcription. © 2013 Elsevier Inc.

Ayer T.,University of Wisconsin - Madison | Chhatwal J.,Merck And Co. | Alagoz O.,University of Wisconsin - Madison | Kahn Jr. C.E.,Medical College of Wisconsin | And 2 more authors.
Radiographics | Year: 2010

Computer models in medical diagnosis are being developed to help physicians differentiate between healthy patients and patients with disease. These models can aid in successful decision making by allowing calculation of disease likelihood on the basis of known patient characteristics and clinical test results. Two of the most frequently used computer models in clinical risk estimation are logistic regression and an artificial neural network. A study was conducted to review and compare these two models, elucidate the advantages and disadvantages of each, and provide criteria for model selection. The two models were used for estimation of breast cancer risk on the basis of mammographic descriptors and demographic risk factors. Although they demonstrated similar performance, the two models have unique characteristics-strengths as well as limitations-that must be considered and may prove complementary in contributing to improved clinical decision making. © RSNA, 2009.

Scher H.I.,Sloan Kettering Cancer Center | Nasso S.F.,Susan G. Komen for the Cure Advocacy Alliance | Rubin E.H.,Merck And Co. | Simon R.,U.S. National Cancer Institute
Clinical Cancer Research | Year: 2011

A critical challenge in the development of new molecularly targeted anticancer drugs is the identification of predictive biomarkers and the concurrent development of diagnostics for these biomarkers. Developing matched diagnostics and therapeutics will require new clinical trial designs and methods of data analysis. The use of adaptive design in phase III trials may offer new opportunities for matched diagnosis and treatment because the size of the trial can allow for subpopulation analysis. We present an adaptive phase III trial design that can identify a suitable target population during the early course of the trial, enabling the efficacy of an experimental therapeutic to be evaluated within the target population as a later part of the same trial. The use of such an adaptive approach to clinical trial design has the potential to greatly improve the field of oncology and facilitate the development of personalized medicine. ©2011 AACR.

Heo M.,Yeshiva University | Pietrobelli A.,University of Verona | Wang D.,Yeshiva University | Heymsfield S.B.,Merck And Co. | Faith M.S.,University of Pennsylvania
Obesity | Year: 2010

To examine the relationship between obesity and functional impairment and the influence of comorbidity, joint pain, and mental health on this association, we used US adult respondents (N = 430,912) to the 2007 Behavioral Risk Factor Surveillance Survey (BRFSS-07). Functional impairment was indicated if a respondent was either (i) limited in any way or in any activities because of physical, mental, or emotional problems, or (ii) had any health problem that required using special equipment such as a cane, wheelchair, special bed, or special telephone. Approximately 62.8% of respondents were overweight or obese and 20.3% were functionally impaired. The unadjusted relationship between obesity and functional impairment revealed a classical J-shaped pattern with odds ratios (95% confidence interval) compared to the normal weight group: 1.63 (1.54-1.73), 1.22 (1.20-1.25), 1.77 (1.73-1.81), 2.43 (2.36-2.51), and 4.12 (3.97-4.27) for underweight, overweight, obesity class I, II, and III, respectively. Although inclusion of different combinations of sociodemographic and medical covariates substantially attenuated the unadjusted association, the collective inclusion of all covariates in a single model did not eliminate the significant J-shaped association resulting in the following corresponding adjusted odds ratios: 1.19 (1.13-1.25), 1.01 (0.99-1.04), 1.23 (1.19-1.27), 1.38 (1.32-1.44), and 1.92 (1.82-2.02). The attenuation was mostly influenced by medical comorbidity. In conclusion, functional impairment is associated with obesity, primarily due to medical comorbidity conditions. The significant residual association highlights the importance of sustainable obesity prevention and treatment at both the individual and public level as functional impairment can create burdens at individual, familial, and societal levels. © 2010 The Obesity Society.

Bellomo A.,University of Pennsylvania | Celebi-Olcum N.,University of California at Los Angeles | Bu X.,Merck And Co. | Rivera N.,Merck And Co. | And 4 more authors.
Angewandte Chemie - International Edition | Year: 2012

A microscale chemistry improvement engine: A pre-dosed microscale high-throughput experimentation additives platform enables rapid, serendipitous reaction improvement. This platform allowed one chemist to set up 475 experiments and analyze the results using MISER chromatography in a single day, thus resulting in two high-quality catalytic systems for the construction of the title compound 1. Support for a single-electron transfer mechanism was obtained. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Murphy J.T.,Sloan Kettering Cancer Center | Burey A.P.,Sloan Kettering Cancer Center | Beebe A.M.,Merck And Co. | Gu D.,Merck And Co. | And 4 more authors.
Blood | Year: 2014

Immunotherapy for cancer using antibodies to enhance T-cell function has been successful in recent clinical trials. Many molecules that improve activation and effector function of T cells have been investigated as potentialnewtargets forimmunomodulatory antibodies, including the tumor necrosis factor receptor superfamily members GITR and OX40. Antibodies engaging GITR or OX40 result in significant tumor protection in preclinical models. In this study, we observed that the GITR agonist antibody DTA-1 causes anaphylaxis in mice upon repeated intraperitoneal dosing. DTA-1-induced anaphylaxis requires GITR, CD4+ T cells, B cells, and interleukin-4. Transfer of serum antibodies from DTA-1-treated mice, which contain high levels of DTA-1-specific immunoglobulin G1 (IgG1), can induce anaphylaxis in naive mice upon administration of an additional dose of DTA-1, suggesting that anaphylaxis results from anti-DTA-1 antibodies. Depletion of basophils and blockade of platelet-activating factor, the key components of the IgG1 pathway of anaphylaxis, rescues themice fromDTA-1-induced anaphylaxis. These results demonstrate a previously undescribed lethal side effect of repetitive doses of an agonist immunomodulatory antibody as well as insight into the mechanism of toxicity, which may offer a means of preventing adverse effects in future clinical trials using anti-GITR or other agonist antibodies as immunotherapies. © 2014 by The American Society of Hematology.

Aschner P.,Pontifical Xavierian University | Katzeff H.L.,Merck And Co. | Guo H.,Merck And Co. | Sunga S.,Merck And Co. | And 3 more authors.
Diabetes, Obesity and Metabolism | Year: 2010

Aim: To compare the efficacy and safety of monotherapy with sitagliptin and metformin in treatment-naïve patients with type 2 diabetes.Methods: In a double-blind study, 1050 treatment-naïve patients (i.e. not taking an antihyperglycaemic agent for ≥16 weeks prior to study entry) with type 2 diabetes and an HbA1c 6.5-9% were randomized (1:1) to treatment with once-daily sitagliptin 100 mg (N = 528) or twice-daily metformin 1000 mg (N = 522) for 24 weeks. Metformin was up-titrated from 500 to 2000 mg per day (or maximum tolerated daily dose ≥1000 mg) over a period of 5 weeks. The primary analysis used a per-protocol (PP) approach to assess whether sitagliptin was non-inferior to metformin based on HbA1c change from baseline at week 24. Non-inferiority was to be declared if the upper boundary of the 95% confidence interval (CI) for the between-group difference in this endpoint was <0.40%.Results: From a mean baseline HbA1c of 7.2% in the PP population, HbA1c change from baseline was -0.43% with sitagliptin (n = 455) and -0.57% with metformin (n = 439). The between-group difference (95% CI) was 0.14% (0.06, 0.21), thus confirming non-inferiority. Baseline HbA1c influenced treatment response, with larger reductions in HbA1c observed in patients with baseline HbA1c ≥8% in the sitagliptin (-1.13%; n = 74) and metformin (-1.24%; n = 73) groups. The proportions of patients at week 24 with HbA1c values at the goals of <7 or <6.5% were 69 and 34% with sitagliptin and 76 and 39% with metformin, respectively. Fasting plasma glucose changes from baseline were -11.5 mg/dL (-0.6 mmol/l) and -19.4 mg/dl (-1.1 mmol/l) with sitagliptin and metformin, respectively (difference in LS mean change from baseline [95% CI] = 8.0 mg /dl [4.5,11.4]). Both treatments led to similar improvements from baseline in measures of homeostasis model assessment-β cell function (HOMA-β) and insulin resistance (HOMA-IR). The incidence of hypoglycaemia was 1.7% with sitagliptin and 3.3% with metformin (p = 0.116). The incidence of gastrointestinal-related adverse experiences was substantially lower with sitagliptin (11.6%) compared with metformin (20.7%) (difference in incidence [95% CI] = -9.1% [-13.6,-4.7]), primarily because of significantly decreased incidences of diarrhoea (3.6 vs. 10.9%; p < 0.001) and nausea (1.1 vs. 3.1%; p = 0.032). Body weight was reduced from baseline with both sitagliptin (LS mean change [95% CI] = -0.6 kg [-0.9,-0.4]) and metformin (-1.9 kg [-2.2, -1.7]) (p < 0.001 for sitagliptin vs. metformin).Conclusions: In this 24-week monotherapy study, sitagliptin was non-inferior to metformin in improving HbA1c in treatment-naïve patients with type 2 diabetes. Although both treatments were generally well tolerated, a lower incidence of gastrointestinal-related adverse experiences was observed with sitagliptin. © 2009 Blackwell Publishing Ltd.

Celanire S.,Domaine de Chosal | Sebhat I.,Merck And Co. | Wichmann J.,Roche Holding AG | Mayer S.,DOMAIN Therapeutics | And 2 more authors.
Expert Opinion on Therapeutic Patents | Year: 2015

Introduction: This review focuses on the medicinal chemistry efforts directed toward the identification of competitive and noncompetitive antagonists of glutamate at group II metabotropic glutamate receptors (mGluRII: mGlu2/3 and mGlu2). This class of compounds holds promise for the treatment of CNS disorders such as major depression, cognitive deficits and sleep-wake disorders, and several pharmaceutical companies are advancing mGluRII antagonists from discovery research into clinical development. Area covered: This review article covers for the first time the patent applications that were published on mGlu2/3 orthosteric and allosteric antagonists between January 2005 and September 2014, with support from the primary literature, posters and oral communications from international congresses. Patent applications published prior to 2005 for which compositions of matter were largely described in peer review articles are briefly discussed with main findings. Expert opinion: Recent advances in the prodrug approach of novel mGlu2/3 orthosteric antagonists combined with the design of novel mGlu2/3 and mGlu2 negative allosteric modulators provide new therapeutic opportunities for neurologic and psychiatric disorders. © 2015 Informa UK, Ltd.

Zhang J.,University of Pennsylvania | Bellomo A.,University of Pennsylvania | Creamer A.D.,University of Pennsylvania | Dreher S.D.,Merck And Co. | Walsh P.J.,University of Pennsylvania
Journal of the American Chemical Society | Year: 2012

Although metal-catalyzed direct arylation reactions of non- or weakly acidic C-H bonds have recently received much attention, chemists have relied heavily on substrates with appropriately placed directing groups to steer reactivity. To date, examples of intermolecular arylation of unactivated C(sp3)-H bonds in the absence of a directing group remain scarce. We report herein the first general, high-yielding, and scalable method for palladium-catalyzed C(sp3)-H arylation of simple diarylmethane derivatives with aryl bromides at room temperature. This method facilitates access to a variety of sterically and electronically diverse hetero- and nonheteroaryl-containing triarylmethanes, a class of compounds with various applications and interesting biological activity. Key to the success of this approach is an in situ metalation of the substrate via C-H deprotonation under catalytic cross-coupling conditions, which is referred to as a deprotonative-cross-coupling process (DCCP). Base and catalyst identification were performed by high-throughput experimentation (HTE) and led to a unique base/catalyst combination [KN(SiMe3)2/Pd-NiXantphos] that proved to efficiently promote the room-temperature DCCP of diarylmethanes. Additionally, the DCCP exhibits remarkable chemoselectivity in the presence of substrates that are known to undergo O-, N-, enolate-, and C(sp2)-H arylation. © 2012 American Chemical Society.

Bifulco G.,University of Salerno | Riccio R.,University of Salerno | Martin G.E.,Merck And Co. | Buevich A.V.,Merck And Co. | Williamson R.T.,Merck And Co.
Organic Letters | Year: 2013

Quantum chemical calculations of one-bond carbon-carbon coupling constants are demonstrated as potential probes for the configurational assignment of organic molecules. The stereochemical analysis of strychnine and its possible stereoisomers is presented as proof of concept. © 2013 American Chemical Society.

Schmink J.R.,University of Pennsylvania | Tudge M.T.,Merck And Co.
Tetrahedron Letters | Year: 2013

A palladium-catalyzed cross coupling of nitrogen bearing heterocyclic chloromethyl derivatives with aryl and heteroaryl boronic acids has been developed. In almost all cases, highly efficient cross-couplings were observed at ambient temperature, mitigating unwanted thermally induced side-reactions. The comprehensive substrate scope and respectable yields highlight the synthetic utility of this reaction. © 2012 Elsevier Ltd. All rights reserved.

Fleury-Bregeot N.,University of Pennsylvania | Raushel J.,University of Pennsylvania | Sandrock D.L.,University of Pennsylvania | Dreher S.D.,Merck And Co. | Molander G.A.,University of Pennsylvania
Chemistry - A European Journal | Year: 2012

Ammoniomethyl trifluoroborates are very powerful reagents that can be used to access biologically relevant aryl- and heteroaryl-methylamine motifs via Suzuki-Miyaura cross-couplings. Until now, this method was limited to the production of tertiary and primary amines. The synthesis of a large array of secondary ammoniomethyltrifluoroborates has been achieved through a one step nucleophilic substitution reaction on the potassium bromomethyltrifluoroborate. Smooth cross-coupling conditions have been designed, based on the use of an aminobiphenyl palladium precatalyst, to couple these trifluoroborates efficiently with aryl bromides. This strategy offers a new way to access biologically relevant motifs and allows, with the previously developed methods, access to all three classes of aminomethylarenes. Secondary ammoniomethyltrifluoroborates can be easily synthesized by nucleophilic substitution on potassium bromomethyltrifluoroborate. These reagents have then been used in Suzuki-Miyaura cross-couplings with aryl bromides, offering an effective access to the aminomethylarene structural motif. This new method provides an interesting alternative to the reductive amination procedure (see scheme). Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Silva A.M.,University of Lisbon | Fields D.A.,University of Oklahoma | Heymsfield S.B.,Merck And Co. | Sardinha L.B.,University of Lisbon
International Journal of Sports Medicine | Year: 2010

The purpose of this study was to analyse the association between body composition changes, from a weight stable period to prior competition, on upper-body power in judo athletes. 27 top-level male athletes were evaluated at baseline (weight stable period) and 1-3 days before competition, with a time difference of approximately 1 month. Total body and extracellular water were estimated by dilution techniques (deuterium and bromide, respectively) and intracellular water was calculated as the difference. Body composition was assessed by DXA. A power-load spectrum was used to assess upper-body power output in a bench-press position. Comparison of means, bivariate, and partial correlations were used. Results indicate that though no significant mean changes were found in body composition and upper-body power, individual variability was large. Among all body composition changes, only total-body water (r=0.672; p<0.001) and intracellular water (r=0.596; p=0.001) were related to upper-body power variation. These associations remained significant after controlling for weight and arm lean-soft tissue changes (r=0.594, p=0.002 for total-body water; r=0.524, p=0.007 for intracellular water). These findings highlight the need for tracking total-body water, specifically the intracellular compartment in elite judo athletes in order to avoid reductions in upper-body power when a target body weight is desired prior to competition. © Georg Thieme Verlag KG Stuttgart - New York.

Gerrits M.G.F.,Merck And Co. | Schnabel P.G.,Merck And Co. | Post T.M.,Merck And Co. | Peeters P.A.M.,Merck And Co. | Peeters P.A.M.,Center for Human Drug Research
Contraception | Year: 2013

Background: The pharmacokinetics of the monophasic oral contraceptive nomegestrol acetate (NOMAC) plus 17β-estradiol (E2) were investigated after a single dose and multiple dosing. Study design: NOMAC/E 2 (2.5 mg/1.5 mg) was administered daily to healthy women (18-50 years, n= 23) for 24 days; blood samples for pharmacokinetic analysis were obtained on Day 24 and again, after a 10-day pill-free interval, on Day 35 after a single dose. Results: NOMAC reached steady state after 5 days with mean ± standard deviation (SD) trough NOMAC concentration (Cav) of 4.4±1.4 ng/mL. On Day 24, mean±SD peak NOMAC concentration (C max, 12.3±3.5 ng/mL) was reached in mean 1.5 h (t max); the mean±SD elimination half-life (t) was 45.9±15.3 h. After a single dose, NOMAC mean±SD Cmax was 7.2±2.0 ng/mL and mean±SD t was 41.9±16.2 h. On Day 24, E2 mean±SD Cav was 50.3±25.7 pg/mL; mean±SD Cmax was 86.0±51.3 pg/mL. After a single dose, mean±SD E2 Cmax was 253±179 pg/mL. Conclusions: These data demonstrate that NOMAC/E2 has a pharmacokinetic profile consistent with once-daily dosing. © 2013 Elsevier Inc.

Kullar R.,Merck And Co. | Sakoulas G.,University of California at San Diego | Sakoulas G.,Sharp Rees Stealy Medical Group | Deresinski S.,Stanford University | Van Hal S.J.,Royal Prince Alfred Hospital
Journal of Antimicrobial Chemotherapy | Year: 2016

MRSA bacteraemia (MRSAB), including infective endocarditis, carries a high mortality rate, with up to 50% of patients failing initial therapy with vancomycin and requiring salvage therapy. Persistent MRSAB can be difficult to successfully eliminate, especially when source control is not possible due to an irremovable focus or the bacteraemia still persists despite surgical intervention. Although vancomycin and daptomycin are the only two antibiotics approved by the US FDA for the treatment of patients with MRSAB as monotherapy, the employment of novel strategies is required to effectively treat patients with persistent MRSAB and these may frequently involve combination drug therapy. Treatment strategies that are reviewed in this manuscript include vancomycin combined with a β-lactam, daptomycin-based therapy, ceftaroline-based therapy, linezolid-based therapy, quinupristin/dalfopristin, telavancin, trimethoprim/sulfamethoxazole-based therapy and fosfomycin-based therapy. We recommend that combination antibiotic therapy be considered for use in MRSAB salvage treatment. © The Author 2015. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved.

Molander G.A.,University of Pennsylvania | Argintaru O.A.,University of Pennsylvania | Aron I.,University of Pennsylvania | Dreher S.D.,Merck And Co.
Organic Letters | Year: 2010

A method for the cross-coupling of alkyl electrophiles with various potassium aryl- and heteroaryltrifluoroborates has been developed. Nearly stoichiometric amounts of organoboron species could be employed to cross-couple a large variety of challenging heteroaryl nucleophiles. Several functional groups were tolerated on both the electrophilic and the nucleophilic partners. Chemoselective reactivity of C(sp3) - Br bonds in the presence of C(sp2) - Br bonds was achieved. © 2010 American Chemical Society.

Murphy S.A.,Harvard University | Cannon C.P.,Harvard University | Blazing M.A.,Duke Clinical Research Institute | Giugliano R.P.,Harvard University | And 11 more authors.
Journal of the American College of Cardiology | Year: 2016

Background Intensive low-density lipoprotein cholesterol therapy with ezetimibe/simvastatin in IMPROVE-IT (IMProved Reduction of Outcomes: Vytorin Efficacy International Trial) significantly reduced the first primary endpoint (PEP) in patients post-acute coronary syndrome (ACS) compared to placebo/simvastatin. Objectives This analysis tested the hypothesis that total events, including those beyond the first event, would also be reduced with ezetimibe/simvastatin therapy. Methods All PEP events (cardiovascular [CV] death, myocardial infarction [MI], stroke, unstable angina [UA] leading to hospitalization, coronary revascularization ≥30 days post-randomization) during a median 6-year follow-up were analyzed in patients randomized to receive ezetimibe/simvastatin or placebo/simvastatin in IMPROVE-IT. Negative binomial regression was used for the primary analysis. Results Among 18,144 patients, there were 9,545 total PEP events (56% were first events and 44% subsequent events). Total PEP events were significantly reduced by 9% with ezetimibe/simvastatin vs placebo/simvastatin (incidence-rate ratio [RR]: 0.91; 95% confidence interval [CI]: 0.85 to 0.97; p = 0.007), as were the 3 pre-specified secondary composite endpoints and the exploratory composite endpoint of CV death, MI, or stroke (RR: 0.88; 95% CI: 0.81 to 0.96; p = 0.002). The reduction in total events was driven by decreases in total nonfatal MI (RR: 0.87; 95% CI: 0.79 to 0.96; p = 0.004) and total NF stroke (RR: 0.77; 95% CI: 0.65 to 0.93; p = 0.005). Conclusions Lipid-lowering therapy with ezetimibe plus simvastatin improved clinical outcomes. Reductions in total PEP events, driven by reductions in MI and stroke, more than doubled the number of events prevented compared with examining only the first event. These data support continuation of intensive combination lipid-lowering therapy after an initial CV event. (IMProved Reduction of Outcomes: Vytorin Efficacy International Trial [IMPROVE-IT]; NCT00202878) © 2016 American College of Cardiology Foundation.

Virchow J.C.,University of Rostock | Backer V.,Copenhagen University | Kuna P.,Medical University of Lódz | Prieto L.,University of Valencia | And 5 more authors.
JAMA - Journal of the American Medical Association | Year: 2016

IMPORTANCE: The house dust mite (HDM) sublingual allergen immunotherapy (SLIT) tablet is a potential novel treatment option for HDM allergy-related asthma. OBJECTIVES To evaluate the efficacy and adverse events of the HDM SLIT tablet vs placebo for asthma exacerbations during an inhaled corticosteroid (ICS) reduction period. DESIGN, SETTINGS, AND PARTICIPANTS: Double-blind, randomized, placebo-controlled trial conducted between August 2011 and April 2013 in 109 European trial sites. The trial included 834 adults with HDM allergy-related asthma not well controlled by ICS or combination products, and with HDM allergy-related rhinitis. Key exclusion criteria were FEV1 less than 70%of predicted value or hospitalization due to asthma within 3 months before randomization. Efficacy was assessed during the last 6 months of the trial when ICS was reduced by 50% for 3 months and then completely withdrawn for 3 months. INTERVENTIONS: 1:1:1 randomization to once-daily treatment with placebo (n = 277) or HDM SLIT tablet (dosage groups: 6 SQ-HDM [n = 275] or 12 SQ-HDM [n = 282]) in addition to ICS and the short-acting β2-agonist salbutamol. MAIN OUTCOMES AND MEASURES: Primary outcomewas time to first moderate or severe asthma exacerbation during the ICS reduction period. Secondary outcomes were deterioration in asthma symptoms, change in allergen-specific immunoglobulin G4 (IgG4), change in asthma control or asthma quality-of-life questionnaires, and adverse events. RESULTS: Among 834 randomized patients (mean age, 33 years [range, 17-83];women, 48%), 693 completed the study. The 6SQ-HDMand 12SQ-HDMdoses both significantly reduced the risk of a moderate or severe asthma exacerbation compared with placebo (hazard ratio [HR]:0.72 [95%CI, 0.52-0.99] for the 6SQ-HDMgroup, P =.045, and 0.69 [95%CI, 0.50-0.96] for the 12 SQ-HDMgroup, P =.03). The absolute risk differences based on the observed data (full analysis set) in the active groups vs the placebo groupwere0.09 (95%CI, 0.01-0.15) for the 6SQ-HDM group and 0.10 (95%CI, 0.02-0.16) for the 12SQ-HDMgroup. Therewas no significant difference between the 2 active groups. Compared with placebo, therewas a reduced risk of an exacerbation with deterioration in asthma symptoms (HR, 0.72 [95%CI, 0.49-1.02] for the 6SQ-HDM group, P =.11, and 0.64 [95%CI, 0.42-0.96] for the 12SQ-HDMgroup, P =.03) and a significant increase in allergen-specific IgG4. However, therewas no significant difference for change in asthma control questionnaire or asthma quality-of-life questionnaire for either dose. Therewere no reports of severe systemic allergic reactions. The most frequent adverse eventswere mild to moderate oral pruritus (13%for the 6SQ-HDMgroup, 20%for the 12SQ-HDMgroup, and 3%for the placebo group), mouth edema, and throat irritation. CONCLUSIONS AND RELEVANCE: Among adults with HDM allergy-related asthma not well controlled by ICS, the addition of HDM SLIT to maintenance medications improved time to first moderate or severe asthma exacerbation during ICS reduction, with an estimated absolute reduction at 6 months of 9 to 10 percentage points; the reduction was primarily due to an effect on moderate exacerbations. Treatment-related adverse events were common at both active doses. Further studies are needed to assess long-term efficacy and safety. © 2016 American Medical Association. All rights reserved.

Sandrock D.L.,University of Pennsylvania | Jean-Gerard L.,University of Pennsylvania | Chen C.-Y.,Merck And Co. | Dreher S.D.,Merck And Co. | Molander G.A.,University of Pennsylvania
Journal of the American Chemical Society | Year: 2010

The stereospecific cross-coupling of enantioenriched nonbenzylic secondary alkyl boron compounds has been achieved. The high selectivity toward product formation over an undesired β-H elimination pathway is achieved via an intramolecular coordination of an ancillary carbonyl to the metal center in the diorganopalladium intermediate. © 2010 American Chemical Society.

Park H.-K.,University of Pennsylvania | Qatanani M.,University of Pennsylvania | Qatanani M.,Merck And Co. | Briggs E.R.,University of Pennsylvania | And 2 more authors.
Diabetes | Year: 2011

OBJECTIVE - Although adipocyte-derived murine resistin links insulin resistance to obesity, the role of human resistin, predominantly expressed in mononuclear cells and induced by inflammatory signals, remains unclear. Given the mounting evidence that obesity and type 2 diabetes are inflammatory diseases, we sought to determine the relationship between inflammatory increases in human resistin and insulin resistance. RESEARCH DESIGN AND METHODS - To investigate the role of human resistin on glucose homeostasis in inflammatory states, we generated mice lacking murine resistin but transgenic for a bacterial artificial chromosome containing human resistin (BAC-Retn), whose expression was similar to that in humans. The metabolic and molecular phenotypes of BAC-Retn mice were assessed after acute and chronic endotoxemia (i.e., exposure to inflammatory lipopolysaccharide). RESULTS - We found that BAC-Retn mice have circulating resistin levels within the normal human range, and similar to humans, lipopolysaccharide markedly increased serum resistin levels. Acute endotoxemia caused hypoglycemia in mice lacking murine resistin, and this was attenuated in BAC-Retn mice. In addition, BAC-Retn mice developed severe hepatic insulin resistance under chronic endotoxemia, accompanied by increased inflammatory responses in liver and skeletal muscle. CONCLUSIONS - These results strongly support the role of human resistin in the development of insulin resistance in inflammation. Thus, human resistin may link insulin resistance to inflammatory diseases such as obesity, type 2 diabetes, and atherosclerosis. © 2011 by the American Diabetes Association.

Bates S.E.,U.S. National Cancer Institute | Berry D.A.,University of Houston | Balasubramaniam S.,Center for Drug Evaluation and Research | Bailey S.,Novartis | And 2 more authors.
Clinical Cancer Research | Year: 2015

The last decade in oncology has been marked by the identification of numerous new potential cancer targets and even more agents designed to inhibit them. The matrix of new targets, new agents, and the companion diagnostics required to identify the right patient for the right drug has created a major challenge for the clinical trial process. This has been compounded by the addition of new immunomodulators targeting the host immune system rather than the tumor. Recognizing the need for new approaches, industry, investigators, and regulators have responded to this challenge. New clinical trial designs are being evaluated to incorporate the genomic sequence data being obtained almost routinely after cancer diagnosis. New dose-finding approaches are being proposed to identify the maximum effective dose rather than the maximum tolerated dose. The FDA is involved in the drug approval process from points early in development and has accepted registration quality data from expansion cohorts in support of drug approval. Despite progress on several fronts, many challenges remain, including the lack of predictability of preclinical data for clinical results and phase II data for phase III results, an infrastructure that can be an obstacle to clinical trial development and implementation, and the increasing use of contracted clinical research organizations that limit a fit-for-purpose approach to clinical trial execution. Perhaps most challenging and important of all are the difficulties with clinical trial accrual that can prevent study completion. Both the innovations and the challenges highlight the important role of process in progress in clinical oncology. © 2015 American Association for Cancer Research.

Trost B.M.,Stanford University | O'Boyle B.M.,Merck And Co. | Hund D.,Albert Ludwigs University of Freiburg
Chemistry - A European Journal | Year: 2010

(Figure Presented) Unexpected Heck: During efforts toward the synthesis of FR900482, an unexpected domino Heck product was isolated (see scheme). Although this pathway was ultimately not productive for the aforementioned natural product synthesis, efforts were made to extend the methodology to the synthesis of amurensinine. To our knowledge this domino reaction was the first such tandem Heck-alkylation pathway in the presence of a geometrically favorable β-hydrogen elimination. © 2010 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Molander G.A.,University of Pennsylvania | Trice S.L.J.,University of Pennsylvania | Kennedy S.M.,University of Pennsylvania | Dreher S.D.,Merck And Co. | Tudge M.T.,Merck And Co.
Journal of the American Chemical Society | Year: 2012

The Suzuki-Miyaura reaction has become one of the more useful tools for synthetic organic chemists. Until recently, there did not exist a direct way to make the most important component in the coupling reaction, namely the boronic acid. Current methods to make boronic acids often employ harsh or wasteful reagents to prepare boronic acid derivatives and require additional steps to afford the desired boronic acid. The scope of the previously reported palladium-catalyzed, direct boronic acid synthesis is unveiled, which includes a wide array of synthetically useful aryl electrophiles. It makes use of the newly available second generation Buchwald XPhos preformed palladium catalyst and bis-boronic acid. For ease of isolation and to preserve the often sensitive C-B bond, all boronic acids were readily converted to their more stable trifluoroborate counterparts. © 2012 American Chemical Society.

Rubin E.H.,Merck And Co. | Allen J.D.,Friends of Cancer Research | Nowak J.A.,NorthShore University Health System Pathology and Laboratory Medicine | Bates S.E.,U.S. National Cancer Institute
Clinical Cancer Research | Year: 2014

Advances in understanding the biology of cancer, as well as advances in diagnostic technologies, such as the advent of affordable high-resolution DNA sequencing, have had a major impact on the approach to identification of specific alterations in a given patient's cancer that could be used as a basis for treatment selection, and hence the development of companion diagnostics. Although there are now several examples of successful development of companion diagnostics that allow identification of patients who will achieve the greatest benefit from a new therapeutic, the path to coapproval of a diagnostic test along with a new therapeutic is complex and often inefficient. This review and the accompanying articles examine the current state of companion diagnostic development in the United States and Europe from academic, industry, regulatory, and economic perspectives.©2014 American Association for Cancer Research.

Herlogsson L.,Linköping University | Colle M.,Merck And Co. | Colle M.,Evonik Industries | Tierney S.,Merck And Co. | And 2 more authors.
Advanced Materials | Year: 2010

A polyanionic electrolyte is used as gate insulator in top-gate p-channel polymer thin-film transistors. The high capacitance of the polyelectrolyte film allows the transistors and integrated circuits to operate below 1.5 V. Seven-stage ring oscillators that operate at supply voltages down to 0.9 V and exhibit signal propagation delays as low as 300 s per stage are reported. © 2010 WILEY-VCH Verlag GmbH & Co. KGaA.

McCurley A.,Molecular Cardiology Research Institute | Pires P.W.,Michigan State University | Bender S.B.,University of Missouri | Aronovitz M.,Nova Southeastern University | And 8 more authors.
Nature Medicine | Year: 2012

Hypertension is a cardiovascular risk factor present in over two-thirds of people over age 60 in North America; elevated blood pressure correlates with increased risk of heart attack, stroke and progression to heart and kidney failure. Current therapies are insufficient to control blood pressure in almost half of these patients. The mineralocorticoid receptor (MR), acting in the kidney, is known to regulate blood pressure through aldosterone binding and stimulation of sodium retention. However, recent studies support the concept that the MR also has extrarenal actions and that defects in sodium handling alone do not fully explain the development of hypertension and associated cardiovascular mortality. We and others have identified functional MR in human vascular smooth muscle cells (SMCs), suggesting that vascular MR might directly regulate blood pressure. Here we show that mice with SMC-specific deficiency of the MR have decreased blood pressure as they age without defects in renal sodium handling or vascular structure. Aged mice lacking MR in SMCs (SMC-MR) have reduced vascular myogenic tone, agonist-dependent contraction and expression and activity of L-type calcium channels. Moreover, SMC-MR contributes to angiotensin II-induced vascular oxidative stress, vascular contraction and hypertension. This study identifies a new role for vascular MR in blood pressure control and in vascular aging and supports the emerging hypothesis that vascular tone contributes directly to systemic blood pressure. © 2012 Nature America, Inc. All rights reserved.

Burlina F.,CNRS Biomolecules Laboratory | Papageorgiou G.,National Insitiute for Medical Research | Morris C.,National Insitiute for Medical Research | White P.D.,Merck And Co. | Offer J.,National Insitiute for Medical Research
Chemical Science | Year: 2014

The progress of total chemical protein synthesis has been hampered by difficulties in preparing peptide thioesters by standard Fmoc peptide synthesis. The amino acid, α-methylcysteine, sited at the C-terminus of a peptide can substitute for a thioester in peptide ligation reactions. C-terminal α-methylcysteine is fully compatible with Fmoc peptide synthesis and its use in ligation is very simple and robust. Its potential is demonstrated with the synthesis of model proteins. © 2014 The Royal Society of Chemistry.

Molander G.A.,University of Pennsylvania | Trice S.L.J.,University of Pennsylvania | Dreher S.D.,Merck And Co.
Journal of the American Chemical Society | Year: 2010

Although much current research focuses on developing new boron reagents and identifying robust catalytic systems for the cross-coupling of these reagents, the fundamental preparations of the nucleophilic partners (i.e., boronic acids and derivatives) has been studied to a lesser extent. Most current methods to access boronic acids are indirect and require harsh conditions or expensive reagents. A simple and efficient palladium-catalyzed, direct synthesis of arylboronic acids from the corresponding aryl chlorides using an underutilized reagent, tetrahydroxydiboron B 2(OH) 4, is reported. To ensure preservation of the carbon-boron bond, the boronic acids were efficiently converted to the trifluoroborate derivatives in good to excellent yields without the use of a workup or isolation. Further, the intermediate boronic acids can be easily converted to a wide range of useful boronates. Finally, a two-step, one-pot method was developed to couple two aryl chlorides efficiently in a Suzuki-Miyaura-type reaction. © 2010 American Chemical Society.

Zeuzem S.,Goethe University Frankfurt | Ghalib R.,The Texas Institute | Reddy K.R.,University of Pennsylvania | Pockros P.J.,Scripps Research Institute | And 10 more authors.
Annals of Internal Medicine | Year: 2015

Background: Novel interferon-and ribavirin-free regimens are needed to treat hepatitis C virus (HCV) infection. Objective: To evaluate the safety and efficacy of grazoprevir (NS3/4A protease inhibitor) and elbasvir (NS5A inhibitor) in treatment-naive patients. Design: Randomized, blinded, placebo-controlled trial. (ClinicalTrials.gov: NCT02105467) Setting: 60 centers in the United States, Europe, Australia, Scandinavia, and Asia. Patients: Cirrhotic and noncirrhotic treatment-naive adults with genotype 1, 4, or 6 infection. Intervention: Oral, once-daily, fixed-dose grazoprevir 100 mg/elbasvir 50 mg for 12 weeks, stratified by fibrosis and genotype. Patients were randomly assigned 3:1 to immediate or deferred therapy. Measurements: Proportion of patients in the immediate-treatment group achieving unquantifiable HCV RNA 12 weeks after treatment (SVR12); adverse events in both groups. Results: Among 421 participants, 194 (46%) were women, 157 (37%) were nonwhite, 382 (91%) had genotype 1 infection, and 92 (22%) had cirrhosis. Of 316 patients receiving immediate treatment, 299 of 316 (95% [95% CI, 92% to 97%]) achieved SVR12, including 144 of 157 (92% [CI, 86% to 96%]) with genotype 1a, 129 of 131 (99% [CI, 95% to 100%]) with genotype 1b, 18 of 18 (100% [CI, 82% to 100%]) with genotype 4, 8 of 10 (80% [CI, 44% to 98%]) with genotype 6, 68 of 70 (97% [CI, 90% to 100%]) with cirrhosis, and 231 of 246 (94% [CI, 90% to 97%]) without cirrhosis. Virologic failure occurred in 13 patients (4%), including 1 case of breakthrough infection and 12 relapses, and was associated with baseline NS5A polymorphisms and emergent NS3 or NS5A variants or both. Serious adverse events occurred in 9 (2.8%) and 3 (2.9%) patients in the active and placebo groups, respectively (difference <0.05 percentage point [CI,-5.4 to 3.1 percentage points]); none were considered drug related. The most common adverse events in the active group were headache (17%), fatigue (16%), and nausea (9%). Limitation: The study lacked an active-comparator control group and included relatively few genotype 4 and 6 infections. Conclusion: Grazoprevir-elbasvir achieved high SVR12 rates in treatment-naive cirrhotic and noncirrhotic patients with genotype 1, 4, or 6 infection. This once-daily, all-oral, fixed-combination regimen represents a potent new therapeutic option for chronic HCV infection. © 2015 American College of Physicians.

Mukherjee R.,University of Pennsylvania | Jensen S.T.,University of Pennsylvania | Male F.,University of Pennsylvania | Bittinger K.,University of Pennsylvania | And 4 more authors.
AIDS | Year: 2011

Objective: Our objective was to analyze the pathways leading to resistance of HIV to the integrase (IN) inhibitor raltegravir (RAL). Design: Three HIV-infected individuals exhibiting RAL resistance pathway switching were characterized using longitudinal analysis of viral samples from plasma. Methods: 454/Roche pyrosequencing was used to generate approximately 74000 sequence reads from the integrase coding region. Effects of error were controlled by denoising with Pyronoise, and by comparison to approximately 142000 control reads from HIVNL4-3. Viral lineages were modeled quantitatively using viral serial pathway analysis (vSPA). Results: All three patients showed transitions from the N155H pathway to the Q148H/G140S pathway. Analysis with vSPA revealed complex pathways to the final genotype, probably involving both de-novo mutation and recombination. No reads contained both the N155H and Q148H drug resistance mutations (DRMs), indicating that the double mutant is not a prominent intermediate, consistent with low fitness. To characterize possible drug-resistant variants circulating prior to therapy, we sequenced approximately 70000 reads from samples collected prior to initiating treatment. Although some preexisting drug-resistant variants were detected, N155H, the first major DRM present after initiating RAL therapy, was not detected. Conclusion: The main DRMs are present at very low levels if at all prior to initiating therapy. We also outline general methods for deep sequence analysis of DRMs in longitudinal HIV samples. © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins.

Mehrotra D.V.,Merck And Co. | Li X.,Gilead Sciences | Liu J.,Merck And Co. | Lu K.,Forest Laboratories
Biometrics | Year: 2012

In a typical randomized clinical trial, a continuous variable of interest (e.g., bone density) is measured at baseline and fixed postbaseline time points. The resulting longitudinal data, often incomplete due to dropouts and other reasons, are commonly analyzed using parametric likelihood-based methods that assume multivariate normality of the response vector. If the normality assumption is deemed untenable, then semiparametric methods such as (weighted) generalized estimating equations are considered. We propose an alternate approach in which the missing data problem is tackled using multiple imputation, and each imputed dataset is analyzed using robust regression (M-estimation; Huber, 1973, Annals of Statistics1, 799-821.) to protect against potential non-normality/outliers in the original or imputed dataset. The robust analysis results from each imputed dataset are combined for overall estimation and inference using either the simple Rubin (1987, Multiple Imputation for Nonresponse in Surveys, New York: Wiley) method, or the more complex but potentially more accurate Robins and Wang (2000, Biometrika87, 113-124.) method. We use simulations to show that our proposed approach performs at least as well as the standard methods under normality, but is notably better under both elliptically symmetric and asymmetric non-normal distributions. A clinical trial example is used for illustration. © 2012, The International Biometric Society No Claim to original US government works.

Cannon C.P.,Brigham and Women's Hospital | Shah S.,Merck And Co. | Dansky H.M.,Merck And Co. | Davidson M.,Radiant Research | And 9 more authors.
New England Journal of Medicine | Year: 2010

BACKGROUND: Anacetrapib is a cholesteryl ester transfer protein inhibitor that raises high-density lipoprotein (HDL) cholesterol and reduces low-density lipoprotein (LDL) cholesterol. METHODS: We conducted a randomized, double-blind, placebo-controlled trial to assess the efficacy and safety profile of anacetrapib in patients with coronary heart disease or at high risk for coronary heart disease. Eligible patients who were taking a statin and who had an LDL cholesterol level that was consistent with that recommended in guidelines were assigned to receive 100 mg of anacetrapib or placebo daily for 18 months. The primary end points were the percent change from baseline in LDL cholesterol at 24 weeks (HDL cholesterol level was a secondary end point) and the safety and side-effect profile of anacetrapib through 76 weeks. Cardiovascular events and deaths were prospectively adjudicated. RESULTS: A total of 1623 patients underwent randomization. By 24 weeks, the LDL cholesterol level had been reduced from 81 mg per deciliter (2.1 mmol per liter) to 45 mg per deciliter (1.2 mmol per liter) in the anacetrapib group, as compared with a reduction from 82 mg per deciliter (2.1 mmol per liter) to 77 mg per deciliter (2.0 mmol per liter) in the placebo group (P<0.001) - a 39.8% reduction with anacetrapib beyond that seen with placebo. In addition, the HDL cholesterol level increased from 41 mg per deciliter (1.0 mmol per liter) to 101 mg per deciliter (2.6 mmol per liter) in the anacetrapib group, as compared with an increase from 40 mg per deciliter (1.0 mmol per liter) to 46 mg per deciliter (1.2 mmol per liter) in the placebo group (P<0.001) - a 138.1% increase with anacetrapib beyond that seen with placebo. Through 76 weeks, no changes were noted in blood pressure or electrolyte or aldosterone levels with anacetrapib as compared with placebo. Prespecified adjudicated cardiovascular events occurred in 16 patients treated with anacetrapib (2.0%) and 21 patients receiving placebo (2.6%) (P = 0.40). The prespecified Bayesian analysis indicated that this event distribution provided a predictive probability (confidence) of 94% that anacetrapib would not be associated with a 25% increase in cardiovascular events, as seen with torcetrapib. CONCLUSIONS: Treatment with anacetrapib had robust effects on LDL and HDL cholesterol, had an acceptable side-effect profile, and, within the limits of the power of this study, did not result in the adverse cardiovascular effects observed with torcetrapib. (Funded by Merck Research Laboratories; ClinicalTrials.gov number, NCT00685776.) Copyright © 2010 Massachusetts Medical Society.

Agency: European Commission | Branch: FP7 | Program: CP | Phase: ICT-2011.3.6 | Award Amount: 14.46M | Year: 2011

Organic photovoltaics (OPV) represent the newest generation of technologies in solar power generation, offering the benefits of flexibility, low weight and low cost enabling the development of new consumer nomadic applications and the long term perspective of easy deployment in Building Integrated Photo Voltaics (BIPV) and energy production farms. This is a key opportunity for the EU to further establish its innovation base in alternative energies.The current challenges reside in the combination to increase efficiencies to 8-10% (module level), increase expected lifetime up to 20 years and decrease production costs to 0.7 Eur/Wp, while taking into account the environmental impact and footprint.The key project objectives are to achieve:\tPrinted OPV with high efficiency architectures such as tandem cells and dedicated light management structures\tHigh performance photo active and passive (barrier) materials including process controlled morphology\tSolutions for cost effective flexible substrates, diffusion barriers and conductors\tDeep understanding of the device physics, elucidation of degradation mechanisms and estimate environmental impact of the main materials and processesThe project consortium combines industrial, institutional and academic support to make a significant impact at European and International level, especially on materials and processes while demonstrating their market-relevant implementations. The industrial project partners are well assembled along the supply chain of future OPV-based products, which is an important prerequisite for the creation of significant socio-economic impact of this proposal.

Zhang J.,University of Pennsylvania | Stanciu C.,University of Pennsylvania | Stanciu C.,Merck And Co. | Wang B.,University of Pennsylvania | And 6 more authors.
Journal of the American Chemical Society | Year: 2011

Although the palladium-catalyzed Tsuji-Trost allylic substitution reaction has been intensively studied, there is a lack of general methods to employ simple benzylic nucleophiles. Such a method would facilitate access to "α-2-propenyl benzyl" motifs, which are common structural motifs in bioactive compounds and natural products. We report herein the palladium-catalyzed allylation reaction of toluene-derived pronucleophiles activated by tricarbonylchromium. A variety of cyclic and acyclic allylic electrophiles can be employed with in situ generated (η 6-C 6H 5CHLiR)Cr(CO) 3 nucleophiles. Catalyst identification was performed by high throughput experimentation (HTE) and led to the Xantphos/palladium hit, which proved to be a general catalyst for this class of reactions. In addition to η 6-toluene complexes, benzyl amine and ether derivatives (η 6-C 6H 5CH 2Z)Cr(CO) 3 (Z = NR 2, OR) are also viable pronucleophiles, allowing C-C bond-formation α to heteroatoms with excellent yields. Finally, a tandem allylic substitution/demetalation procedure is described that affords the corresponding metal-free allylic substitution products. This method will be a valuable complement to the existing arsenal of nucleophiles with applications in allylic substitution reactions. © 2011 American Chemical Society.

Schmink J.R.,University of Pennsylvania | Krska S.W.,Merck And Co.
Journal of the American Chemical Society | Year: 2011

Acylsilanes serve as acyl anion equivalents in a palladium-catalyzed cross-coupling reaction with aryl bromides to give unsymmetrical diaryl ketones. Water plays a unique and crucial activating role in these reactions. High-throughput experimentation techniques provided successful reaction conditions initially involving phosphites as ligands. Ultimately, 1,3,5,7-tetramethyl-6-phenyl-2,4,8-trioxa-6-phosphaadamantane was identified as giving a longer-lived catalyst with higher turnover numbers. Its use, in conjunction with a palladacycle precatalyst, led to optimal reaction rates and yields. Scope and limitations of this novel method are presented along with initial mechanistic insight. © 2011 American Chemical Society.

Kraft W.K.,Thomas Jefferson University | Chang P.S.,Thomas Jefferson University | Van Iersel M.L.P.S.,MSD | Waskin H.,Merck And Co. | And 3 more authors.
Antimicrobial Agents and Chemotherapy | Year: 2014

Posaconazole oral suspension is an extended-spectrum triazole that should be taken with food to maximize absorption. A new posaconazole tablet formulation has demonstrated improved bioavailability over the oral suspension in healthy adults in a fasting state. This study evaluated the effects of concomitant medications altering gastric pH (antacid, ranitidine, and esomeprazole) and gastric motility (metoclopramide) on the pharmacokinetics of posaconazole tablets. This was a prospective open-label 5-way crossover study in 20 healthy volunteers. In each treatment period, a single 400-mg dose (4 100-mg tablets) of posaconazole was administered alone or with 20 ml antacid (2 g of aluminum hydroxide and 2 g of magnesium hydroxide), ranitidine (150 mg), esomeprazole (40 mg), or metoclopramide (15 mg). There was a >10-day washout between treatment periods. Posaconazole exposure, time to maximum concentration of drug in serum (Tmax), and apparent terminal half-life (t1/2) were similar when posaconazole was administered alone or with medications affecting gastric pH and gastric motility. Geometric mean ratios (90% confidence intervals [CIs]) of the area under the concentration-time curve from time zero to infinity (AUC0-inf) (posaconazole with medications affecting gastric pH and gastric motility versus posaconazole alone) were 1.03 (0.88-1.20) with antacid, 0.97 (0.84-1.12) with ranitidine, 1.01 (0.87-1.17) with esomeprazole, and 0.93 (0.79-1.09) with metoclopramide. Geometric mean ratios (90% CIs) of the maximum concentration of drug in serum (Cmax) were 1.06 (0.90-1.26) with antacid, 1.04 (0.88-1.23) with ranitidine, 1.05 (0.89-1.24) with esomeprazole, and 0.86 (0.73-1.02) with metoclopramide. In summary, in healthy volunteers, the pharmacokinetics of a single 400-mg dose of posaconazole tablets was not altered to a clinically meaningful extent when posaconazole was administered alone or with medications affecting gastric pH or gastric motility. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Weiss T.W.,Merck And Co. | Zimet G.D.,Indiana University | Rosenthal S.L.,Columbia University | Brenneman S.K.,i3 Innovus | And 2 more authors.
Journal of Adolescent Health | Year: 2010

Purpose: We assessed U.S. physicians' attitudes and perceptions regarding potential human papillomavirus (HPV) vaccination of males. Methods: We surveyed a random sample of 2,714 pediatricians and family practitioners identified in administrative claims of a U.S. health plan as HPV vaccinators of females; 595 pediatricians and 499 family practitioners participated. Results: Most physicians would recommend HPV vaccination to males aged 11-12 (63.9%), 13-18 (93.4%), and 19-26 (92.7%) years. Physicians agreed that males should be vaccinated to prevent them from getting genital and anal warts (52.9% strongly and 36.0% somewhat) and to protect females from cervical cancer (75.3% strongly and 20.8% somewhat). Physicians agreed that an HPV vaccine recommendation for males would increase opportunities to discuss sexual health with adolescent male patients (58.7% strongly, 35.3% somewhat). Most did not strongly agree (15.4% strongly, 45.4% somewhat) that parents of adolescent male patients would be interested in HPV vaccination for males, that a gender-neutral HPV vaccine recommendation would increase acceptance by adolescent females and their parents (19.6% strongly, 42.0% somewhat), or that a gender-neutral recommendation would improve current female vaccination rates (10.4% strongly, 26.0% somewhat). Conclusions: Physicians who currently vaccinate females against HPV supported the concept of vaccinating males for its benefits for both sexes. They agreed that a gender-neutral HPV vaccination recommendation would be appropriate with regard to public health and believed that it would increase opportunities for sexual health discussions, but were less sure that such a recommendation would change patient or parental attitudes toward HPV vaccination or improve current HPV vaccination efforts. © 2010 Society for Adolescent Health and Medicine.

Jacus M.O.,University of Virginia | Uebele V.N.,Merck And Co. | Renger J.J.,Merck And Co. | Todorovic S.M.,University of Virginia
Journal of Neuroscience | Year: 2012

It is generally accepted that presynaptic transmitter release is mainly regulated by subtypes of neuronal high-voltage-activated Ca 2+ channels. Here for the first time, we examined the role of T-type Ca 2+ channels (T-channels) in synaptic transmission in the dorsal horn (DH) of the spinal cord using patch-clamp recordings from acute spinal cord preparations from both rat and mouse. We found that selective pharmacological antagonism of T-channels inhibited spontaneous synaptic release of glutamate in superficial laminae I-II of the DH, while GABA release was spared. We found similar effect in identified nociceptive projection neurons of lamina I of the DH, but not in inhibitory DH interneurons. In comparison, antagonism of T-channels did not affect excitatory transmission in deeper non-nociceptive DH laminae. Furthermore, we used isoform-specific agents, knock-out mice and immunohistochemistry to specifically implicate presynaptic CaV3.2 channels. We also used an animal model of painful diabetic neuropathy to demonstrate that blocking T-channels in superficial DH neurons suppressed spontaneous excitatory synaptic transmission in diabetic rats in greater degree than in healthy age-matched animals. These studies provide previously unknown information regarding the role of presynaptic T-channels in nociceptive signaling in the spinal cord. © 2012 the authors.

Liao J.J.Z.,Teva Pharmaceuticals | Capen R.,Merck And Co.
International Journal of Biostatistics | Year: 2011

It is often necessary to compare two measurement methods in medicine and other experimental sciences. This problem covers a broad range of data with applications arising from many different fields. The Bland-Altman method has been a favorite method for concordance assessment. However, the Bland-Altman approach creates a problem of interpretation for many applications when a mixture of fixed bias, proportional bias and/or proportional error occurs. In this paper, an improved Bland-Altman method is proposed to handle more complicated scenarios in practice. This new approach includes Bland-Altman's approach as its special case. We evaluate concordance by defining an agreement interval for each individual paired observation and assessing the overall concordance. The proposed interval approach is very informative and offers many advantages over existing approaches. Data sets are used to demonstrate the advantages of the new method. © 2011 Berkeley Electronic Press. All rights reserved.

Kumar P.,University of Utah | Kumar P.,Merck And Co. | Thakur A.,University of Utah | Hong X.,University of California at Los Angeles | And 2 more authors.
Journal of the American Chemical Society | Year: 2014

A Ni/N-heterocyclic carbene catalyst couples diynes to the C(α)-C(β) double bond of tropone, a type of reaction that is unprecedented for metal-catalyzed cycloadditions with aromatic tropone. Many different diynes were efficiently coupled to afford [5-6-7] fused tricyclic products, while [5-7-6] fused tricyclic compounds were obtained as minor byproducts in a few cases. The reaction has broad substrate scope and tolerates a wide range of functional groups, and excellent regioselectivity is found with unsymmetrical diynes. Theoretical calculations show that the apparent enone cycloaddition occurs through a distinctive 8π insertion of tropone. The initial intramolecular oxidative cyclization of diyne produces the nickelacyclopentadiene intermediate. This intermediate undergoes an 8π insertion of tropone, and subsequent reductive elimination generates the [5-6-7] fused tricyclic product. This initial product undergoes two competing isomerizations, leading to the observed [5-6-7] and [5-7-6] fused tricyclic products. © 2014 American Chemical Society.

Morrow D.A.,Brigham and Women's Hospital | Braunwald E.,Brigham and Women's Hospital | Bonaca M.P.,Brigham and Women's Hospital | Ameriso S.F.,Institute for Neurological Research | And 14 more authors.
New England Journal of Medicine | Year: 2012

BACKGROUND: Thrombin potently activates platelets through the protease-activated receptor PAR-1. Vorapaxar is a novel antiplatelet agent that selectively inhibits the cellular actions of thrombin through antagonism of PAR-1. METHODS: We randomly assigned 26,449 patients who had a history of myocardial infarction, ischemic stroke, or peripheral arterial disease to receive vorapaxar (2.5 mg daily) or matching placebo and followed them for a median of 30 months. The primary efficacy end point was the composite of death from cardiovascular causes, myocardial infarction, or stroke. After 2 years, the data and safety monitoring board recommended discontinuation of the study treatment in patients with a history of stroke owing to the risk of intracranial hemorrhage. RESULTS:At 3 years, the primary end point had occurred in 1028 patients (9.3%) in the vorapaxar group and in 1176 patients (10.5%) in the placebo group (hazard ratio for the vorapaxar group, 0.87; 95% confidence interval [CI], 0.80 to 0.94; P<0.001). Cardiovascular death, myocardial infarction, stroke, or recurrent ischemia leading to revascularization occurred in 1259 patients (11.2%) in the vorapaxar group and 1417 patients (12.4%) in the placebo group (hazard ratio, 0.88; 95% CI, 0.82 to 0.95; P = 0.001). Moderate or severe bleeding occurred in 4.2% of patients who received vorapaxar and 2.5% of those who received placebo (hazard ratio, 1.66; 95% CI, 1.43 to 1.93; P<0.001). There was an increase in the rate of intracranial hemorrhage in the vorapaxar group (1.0%, vs. 0.5% in the placebo group; P<0.001). CONCLUSIONS:Inhibition of PAR-1 with vorapaxar reduced the risk of cardiovascular death or ischemic events in patients with stable atherosclerosis who were receiving standard therapy. However, it increased the risk of moderate or severe bleeding, including intracranial hemorrhage. (Funded by Merck; TRA 2P-TIMI 50 ClinicalTrials.gov number, NCT00526474.) Copyright © 2012 Massachusetts Medical Society.

Hill A.,Evotec | Mesens N.,Janssen Pharmaceutical | Steemans M.,Janssen Pharmaceutical | Xu J.J.,Merck And Co. | Aleo M.D.,Pfizer
Drug Metabolism Reviews | Year: 2012

Drug-induced liver injury (DILI) is a major cause of attrition during both the early and later stages of the drug development and marketing process. Reducing or eliminating drug-induced severe liver injury, especially those that lead to liver transplants or death, would be tremendously beneficial for patients. Therefore, developing new pharmaceuticals that have the highest margins and attributes of hepatic safety would be a great accomplishment. Given the current low productivity of pharmaceutical companies and the high costs of bringing new medicines to market, any early screening assay(s) to identify and eliminate pharmaceuticals with the potential to cause severe liver injury in humans would be of economic value as well. The present review discusses the background, proof-of-concept, and validation studies associated with high-content screening (HCS) by two major pharmaceutical companies (Pfizer Inc and Jansen Pharmaceutical Companies of Johnson & Johnson) for detecting compounds with the potential to cause human DILI. These HCS assays use fluorescent-based markers of cell injury in either human hepatocytes or HepG2 cells. In collaboration with Evotec, an independent contract lab, these two companies also independently evaluated larval zebrafish as an early-stage in vivo screen for hepatotoxicity in independently conducted, blinded assessments. Details about this model species, the need for bioanalysis, and, specifically, the outcome of the phenotypic-based zebrafish screens are presented. Comparing outcomes in zebrafish against both HCS assays suggests an enhanced detection for hepatotoxicants of most DILI concern when used in combination with each other, based on the U.S. Food and Drug Administration DILI classification list. © 2012 Informa Healthcare USA, Inc.

Aragon E.,Barcelona Institute for Research in Biomedicine | Goerner N.,Barcelona Institute for Research in Biomedicine | Goerner N.,Biocrates Life Sciences | Xi Q.,Sloan Kettering Cancer Center | And 7 more authors.
Structure | Year: 2012

Transforming growth factor (TGF)-β and BMP signaling is mediated by Smads 1-5 (R-Smads and Co-Smads) and inhibited by Smad7, a major hub of regulation of TGF-β and BMP receptors by negative feedback and antagonistic signals. The transcription coactivator YAP and the E3 ubiquitin ligases Smurf1/2 and Nedd4L target R-Smads for activation or degradation, respectively. Pairs of WW domain in these regulators bind PY motifs and adjacent CDK/MAPK and GSK3 phosphorylation sites in R-Smads in a selective and regulated manner. In contrast, here we show that Smad7 binds YAP, Smurf1, Smurf2, and Nedd4L constitutively, the binding involving a PY motif in Smad7 and no phosphorylation. We also provide a structural basis for how regulators that use WW domain pairs for selective interactions with R-Smads, resort to one single versatile WW domain for binding Smad7 to centralize regulation in the TGF-β and BMP pathways. © 2012 Elsevier Ltd.

Price D.,University of Aberdeen | Popov T.A.,Sofia University | Bjermer L.,Skåne University Hospital | Lu S.,Merck And Co. | And 6 more authors.
Journal of Allergy and Clinical Immunology | Year: 2013

Objective: Many asthmatic patients are unable to quit cigarettes; therefore information is needed on treatment options for smokers. This study evaluates 10 mg/d montelukast and 250 μg of fluticasone propionate twice daily, each compared with placebo, in patients with self-reported active smoking (unable to quit) and asthma. Methods: Patients (ages 18-55 years, with asthma [≥1 year], FEV1 of 60% to 90% of predicted value, airway reversibility [≥12%], and self-reported active smoking [≥0.5 to ≤2 packs per day]) were randomized (after a 3-week, single-blind, placebo, run-in period) to 1 of 3 parallel, 6-month, double-blind treatment arms. The primary efficacy end point was the percentage of days with asthma control during treatment. Adverse experiences (AEs) were also evaluated. Results: There were 347, 336, and 336 patients randomized to montelukast, fluticasone, and placebo, respectively. The mean percentage of days with asthma control over 6 months of treatment was 45% (montelukast, P < .05 vs placebo), 49% (fluticasone, P < .001 vs placebo), and 39% (placebo); the difference between montelukast and fluticasone was not significant (P = .14). Patients with a smoking history of ≤11 pack years (the median value) tended to show more benefit with fluticasone, whereas those with a smoking history of >11 pack years tended to show more benefit with montelukast. AEs occurred in similar proportions among treatment groups. Conclusions: In a population of asthmatic patients actively smoking cigarettes, both 10 mg/d montelukast and 250 μg of fluticasone propionate twice daily significantly increased the mean percentage of days with asthma control compared with placebo. © 2013 American Academy of Allergy, Asthma & Immunology.

Van Nas A.,University of California at Los Angeles | Ingram-Drake L.,University of California at Los Angeles | Sinsheimer J.S.,University of California at Los Angeles | Wang S.S.,Merck And Co. | And 3 more authors.
Genetics | Year: 2010

By treating the transcript abundance as a quantitative trait, gene expression can be mapped to local or distant genomic regions relative to the gene encoding the transcript. Local expression quantitative trait loci (eQTL) generally act in cis (that is, control the expression of only the contiguous structural gene), whereas distal eQTL act in trans. Distal eQTL are more difficult to identify with certainty due to the fact that significant thresholds are very high since all regions of the genome must be tested, and confounding factors such as batch effects can produce false positives. Here, we compare findings from two large genetic crosses between mouse strains C3H/HeJ and C57BL/6J to evaluate the reliability of distal eQTL detection, including "hotspots" influencing the expression of multiple genes in trans. We found that >63% of local eQTL and >18% of distal eQTL were replicable at a threshold of LOD > 4.3 between crosses and 76% of local and >24% of distal eQTL at a threshold of LOD > 6. Additionally, at LOD > 4.3 four tissues studied (adipose, brain, liver, and muscle) exhibited >50% preservation of local eQTL and >17% preservation of distal eQTL. We observed replicated distal eQTL hotspots between the crosses on chromosomes 9 and 17. Finally, >69% of local eQTL and >10% of distal eQTL were preserved in most tissues between sexes. We conclude that most local eQTL are highly replicable between mouse crosses, tissues, and sex as compared to distal eQTL, which exhibited modest replicability. Copyright © 2010 by the Genetics Society of America.

Patel U.D.,Duke University | Greiner M.A.,Duke University | Fonarow G.C.,University of California at Los Angeles | Phatak H.,Merck And Co. | And 2 more authors.
American Heart Journal | Year: 2010

Background: Kidney disease is common among patients with heart failure, but relationships between worsening renal function (WRF) and outcomes after hospitalization for heart failure are poorly understood, especially among patients with preserved systolic function. We examined associations between WRF and 30-day readmission, mortality, and costs among Medicare beneficiaries hospitalized with heart failure. Methods: We linked data from a clinical heart failure registry to Medicare inpatient claims for patients ≥65 years old hospitalized with heart failure. We defined WRF as a change in serum creatinine ≥0.3 mg/dL from admission to discharge. Main outcome measures were readmission and mortality at 30 days after hospitalization and total inpatient costs. Results: Among 20,063 patients hospitalized with heart failure, WRF was common (17.8%) and more likely among patients with higher baseline comorbidity and more impaired renal function. In unadjusted analyses, WRF was associated with similar subsequent mean inpatient costs ($3,255 vs $3,277, P = .2) but higher readmission (21.8% vs 20.6%, P = .01) and mortality (10.0% vs 7.2%, P < .001). The differences persisted after adjustment for baseline patient and hospital characteristics (hazard of readmission 1.10 [95% CI 1.02-1.18], hazard of mortality 1.53 [95% CI 1.34-1.75]). Associations of WRF with readmission and mortality were similar between patients with reduced and preserved systolic function. Conclusions: Worsening renal function during hospitalization for heart failure is an independent predictor of early readmission and mortality in patients with reduced and preserved systolic function. © 2010, Mosby, Inc. All rights reserved.

Martin G.E.,Merck And Co. | Hilton B.D.,Merck And Co. | Willcott III M.R.,Rice University | Blinov K.A.,ACD Laboratories
Magnetic Resonance in Chemistry | Year: 2011

Utilizing 13C-13C connectivity networks for the assembly of carbon skeletons from HSQC-ADEQUATE spectra was recently reported. HSQC-ADEQUATE data retain the resonance multiplicity information of the multiplicity-edited GHSQC spectrum and afford a significant improvement in the signal-to-noise (s/n) ratio relative to the 1,1-ADEQUATE data used in the calculation of the HSQC-ADEQUATE spectrum by unsymmetrical indirect covariance (UIC) processing methods. The initial investigation into the computation of HSQC-ADEQUATE correlation plots utilized overnight acquisition of the 1,1-ADEQUATE data used for the calculation. In this communication, we report the results of an investigation of the reduction in acquisition time for the 1,1-ADEQUATE data to take advantage of the s/n gain during the UIC processing to afford the final HSQC-ADEQUATE correlation plot. Data acquisition times for the 1,1-ADEQUATE spectrum can be reduced to as little as a few hours, while retaining excellent s/n ratios and all responses contained in spectra computed from overnight data acquisitions. Concatenation of multiplicity-edited GHSQC and 1,1-ADEQUATE data also allows the interrogation of submilligram samples with 1,1-ADEQUATE data when using spectrometers equipped with 1.7-mm Micro CryoProbes Trademark. © 2011 John Wiley & Sons, Ltd.

Towne V.,Merck And Co. | Zhao Q.,Merck And Co. | Zhao Q.,Xiamen University | Brown M.,Merck And Co. | Finnefrock A.C.,Merck And Co.
Journal of Immunological Methods | Year: 2013

This paper describes an approach to surface plasmon resonance (SPR) based epitope mapping, also referred to as pairwise antibody footprinting, involving the direct immobilization of an antigen-specific primary mAb to the surface of an SPR interface. This technique offers a more straightforward approach than indirect capture (e.g., via rabbit anti-mouse Fc) as it does not require additional steps to block the unoccupied immobilized anti-Fc to prevent non-specific antibody binding. This is also an alternative to the direct immobilization of an antigen of interest, which may cause conformational changes in the antigen or epitope degradation upon chemical immobilization, particularly in successive regeneration cycles. It is particularly suitable for highly multivalent targets such as virus-like particles (VLPs). Using this technique, we assessed a panel of eight monoclonal antibodies against HPV (human papilloma virus) L1 protein VLPs expressed by Saccharomyces cerevisiae. In the antibody epitope screening studies, HPV16 L1-directed conformational mAbs were clearly distinguished from the linear mAbs and consistent with known epitope information. Additional studies using a linear mAb and a conformational mAb demonstrate the practical application of this technique for characterizing the result of process changes and the consistency of recombinant HPV16 VLPs. The method is readily extensible to other VLPs and VLP-based vaccines. © 2012 Elsevier B.V..

Bigal M.E.,Merck And Co. | Bigal M.E.,Yeshiva University | Rapoport A.M.,University of California at Los Angeles
Current Pain and Headache Reports | Year: 2012

Obesity may be the greatest epidemic of modern times. It leads to diabetes and heart disease and shortens lifespan. Although not a risk factor for migraine, it is associated with an increased frequency and intensity of migraine. Obesity is also comorbid with chronic daily headache and is a major risk factor for chronification of episodic migraine in adults and children. Although obesity is not a factor in the effectiveness of migraine treatment, it does increase the peripheral and central events in migraine, ultimately increasing the neurologic potential for migraine. Although evidence suggests that obesity is a modifiable risk factor for migraine progression, it is unknown if weight loss is related to decrease in headache frequency. Recent surgical results suggest that this is true. We suggest all possible effective techniques aimed at weight loss be undertaken for migraineurs, especially obese migraineurs, and that carefully monitoring weight changes should be routinely done as part of their migraine care. © 2011 Springer Science+Business Media, LLC.

Yao J.,Stanley Institute for Cognitive Genomics | Yao J.,Merck And Co. | Zhang K.X.,Howard Hughes Medical Institute | Kramer M.,Stanley Institute for Cognitive Genomics | And 2 more authors.
Bioinformatics | Year: 2014

FamAnn is an automated variant annotation pipeline designed for facilitating target discovery for family-based sequencing studies. It can apply a different inheritance pattern or a de novo mutations discovery model to each family and select single nucleotide variants and small insertions and deletions segregating in each family or shared by multiple families. It also provides a variety of variant annotations and retains and annotates all transcripts hit by a single variant. Excel-compatible outputs including all annotated variants segregating in each family or shared by multiple families will be provided for users to prioritize variants based on their customized thresholds. A list of genes that harbor the segregating variants will be provided as well for possible pathway/network analyses. FamAnn uses the de facto community standard Variant Call Format as the input format and can be applied to whole exome, genome or targeted resequencing data. © 2014 The Author.

Ivanova J.I.,Analysis Group Inc. | Birnbaum H.G.,Analysis Group Inc. | Kidolezi Y.,Analysis Group Inc. | Qiu Y.,Merck And Co. | And 2 more authors.
Epilepsia | Year: 2010

Purpose: Compare annual direct and indirect costs between privately insured U.S. patients with epileptic partial onset seizures (POS) and matched controls. Methods: One thousand eight hundred fifty-nine patients (including a subset of 758 employees) with ≥1 (POS) diagnosis (ICD-9-CM: 345.4.x-345.7.x), 1999-2004, ages 16-64 years, were identified from a privately insured claims database. Control group was an age- and gender-matched cohort of randomly chosen beneficiaries without epilepsy (ICD-9-CM: 345.x). All were required to have continuous health coverage during 2004 (baseline) and 2005 (study period). Chi-square tests were used to compare baseline comorbidities. Univariate and multivariate analyses were used for comparisons of annual direct (medical and pharmaceutical) and indirect costs during the study period. Results: Patients with POS were on average 42 years of age, and 57% were women. Patients with POS had significantly higher rates of mental health disorders, migraine, and other neurologic disorders, and higher Charlson comorbidity index (CCI) compared with controls. On average, direct annual costs were significantly higher for POS patients ($11,276) compared with controls ($4,087), p < 0.001; difference of $7,190. Epilepsy-related costs (i.e., costs for antiepileptic drugs, claims with epilepsy or convulsions diagnoses) accounted for $3,290 (29% of direct costs). Employees with POS had substantial and significantly higher indirect (disability-and medically related absenteeism) costs compared with controls ($3,431 vs. $1,511, p < 0.001). Multivariate analyses supported the matched-control univariate findings. Conclusion: Patients with POS had significantly higher costs compared with matched controls. Epilepsy-related costs underestimate the excess costs of patients with partial onset seizures. © 2009 International League Against Epilepsy.

Ofir-Birin Y.,Hebrew University of Jerusalem | Fang P.,Scripps Research Institute | Bennett S.P.,Scripps Research Institute | Zhang H.-M.,Florida State University | And 14 more authors.
Molecular Cell | Year: 2013

Lysyl-tRNA synthetase (LysRS), a component of the translation apparatus, is released from the cytoplasmic multi-tRNA synthetase complex (MSC) to activate the transcription factor MITF in stimulated mast cells through undefined mechanisms. Here we show that Ser207 phosphorylation provokes a new conformer of LysRS that inactivates its translational function but activates its transcriptional function. The crystal structure of an MSC subcomplex established that LysRS is held in the MSC by binding to the N terminus of the scaffold protein p38/AIMP2. Phosphorylation-created steric clashes at the LysRS domain interface disrupt its binding grooves for p38/AIMP2, releasing LysRS and provoking its nuclear translocation. This alteration also exposes the C-terminal domain of LysRS to bind to MITF and triggers LysRS-directed production of the second messenger Ap4A that activates MITF. Thus our results establish that a single conformational change triggered by phosphorylation leads to multiple effects driving an exclusive switch of LysRS function from translation to transcription. © 2013 Elsevier Inc.

Boyce B.F.,University of Rochester | Rosenberg E.,Merck And Co. | de Papp A.E.,Merck And Co. | Duong L.T.,Merck And Co.
European Journal of Clinical Investigation | Year: 2012

Background Bone remodelling maintains skeletal integrity by osteoclasts removing foci of damaged bone and osteoblasts replacing them with new bone. Diseases associated with increased bone resorption have increased remodelling often with inadequate bone formation and increased risk of fracture. New therapies are needed for these diseases to reduce resorption and increase formation. Design The molecular mechanisms regulating osteoclast and osteoblast functions have become better understood in the past 20years and have led to questioning of the long-held notion that osteoblastic cells have the dominant regulatory role over osteoclastic cells in bone remodelling. Here, we review current knowledge of how osteoclast formation and functions are regulated and describe how enhanced understanding of these has led to development of new drugs for the management of common bone diseases characterized by increased bone resorption. Results Osteoclast formation and functions are regulated by cytokines, especially receptor activator of NF-κB ligand (RANKL) and macrophage-colony-stimulating factor (M-CSF). The differentiation, activity and lifecycle of osteoclasts are regulated in part by other cells that reside within the bone. These include osteoblasts, osteocytes and immune cells, which express these cytokines in response to most factors that promote bone resorption. RANKL and M-CSF activate numerous signalling pathways, which are potential targets for therapeutic intervention. Importantly, osteoclastic cells also function as positive and negative regulators of osteoblastic bone formation. Conclusions There are multiple targets within osteoclasts for pharmacologic intervention to prevent bone loss in osteoporosis and other resorptive bone diseases. However, novel therapies could also affect osteoblastic cell functions. © 2012 Stichting European Society for Clinical Investigation Journal Foundation.

Trigg M.E.,Merck And Co. | Trigg M.E.,Thomas Jefferson University | Higa G.M.,West Virginia University
Journal of Oncology Pharmacy Practice | Year: 2010

Objective. To review the scientific evidence related to serotonin and substance P and the clinical impact targeting these two neurotransmitters have had managing chemotherapy-induced nausea and vomiting (CINV). Data Source. A PubMed search (January 1968 to December 2008), restricted to English-language publications, was conducted using the key words antiemetics, cancer chemotherapy, cisplatin, serotonin, substance P, NK 1, and 5-HT 3. Abstracts emanating from the meetings of the American Society of Clinical Oncology and Multinational Association of Supportive Care in Cancer during the period May 2000 to June 2008 were also reviewed. Data Synthesis. Two important outcomes emanated from well-conducted antiemetic clinical trials (Table 1): first, evidence that serotonin and substance P are major mediators of acute and delayed symptoms and second, improved, though not complete, control of CINV. Conclusion. Serotonin-type 3 and neurokinin-1 receptor antagonists are the most effective agents currently available. In most cases, these agents are used in conjunction with glucocorticoids. The use of these three types of agents is incorporated into current clinical practice guidelines. Further understanding of the biological and biochemical basis of nausea and vomiting may enhance management of this potentially debilitating adverse effect. © The Author(s), 2010.

Maertens J.,University Hospital Gasthuisberg | Cornely O.A.,University of Cologne | Ullmann A.J.,Johannes Gutenberg University Mainz | Heinz W.J.,University of Würzburg | And 6 more authors.
Antimicrobial Agents and Chemotherapy | Year: 2014

This was a phase 1B, dose-ranging, multicenter, pharmacokinetics, and safety study of cyclodextrin-based posaconazole intravenous (i.v.) solution administered through a central line to subjects at high risk for invasive fungal disease (part 1 of a 2-part study [phase 1B/3]). Initially, the safety and tolerability of single-dose posaconazole i.v. 200 mg (n = 10) were compared with those of a placebo (n = 11). Subsequently, 2 doses were evaluated, posaconazole i.v. 200 mg once daily (q.d.) (n = 21) and 300 mg q.d. (n = 24). The subjects received twice-daily (b.i.d.) posaconazole i.v. on day 1, followed by 13 days of posaconazole i.v. q.d., then 14 days of posaconazole oral suspension 400 mg b.i.d. The steady-state (day 14) exposure target (average concentration [areas under concentration-time curve {AUCs}/24 h, average concentrations at steady state {Cavgs}], of ≥500 to ≤2,500 ng/ml in ≥90% of the subjects) was achieved by 94% of the subjects for 200 mg posaconazole q.d. and by 95% of subjects for 300 mg posaconazole q.d. The desired exposure target (mean steady-state Cavg, ∼1,200 ng/ml) was 1,180 ng/ml in the 200-mg dosing cohort and was exceeded in the 300-mg dosing cohort (1,430 ng/ml). Posaconazole i.v. was well tolerated. Posaconazole i.v. 300 mg q.d. was selected for the phase 3 study segment. (This study has been registered at ClinicalTrials.gov under registration no. NCT01075984.) Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Ogilvie I.,TransCanada | Khoury H.,TransCanada | Goetghebeur M.M.,TransCanada | El Khoury A.C.,Merck And Co. | Giaquinto C.,University of Padua
BMC Infectious Diseases | Year: 2012

Background: Rotavirus affects 95% of children worldwide by age 5 years and is the leading cause of severe dehydrating diarrhea. The objective of this review was to estimate the burden of rotavirus gastroenteritis (RVGE) in the Western European pediatric population.Methods: A comprehensive literature search (1999-2010) was conducted in PubMed and other sources (CDC; WHO, others). Data on the epidemiology and burden of RVGE among children < 5 years-old in Western Europe --including hospital-acquired disease--were extracted.Results: 76 studies from 16 countries were identified. The mean percentage of acute gastroenteritis (AGE) cases caused by rotavirus ranged from 25.3%-63.5% in children < 5 years of age, peaking during winter. Incidence rates of RVGE ranged from 1.33-4.96 cases/100 person- years. Hospitalization rates for RVGE ranged from 7% to 81% among infected children, depending on the country. Nosocomial RVGE accounted for 47%-69% of all hospital-acquired AGE and prolonged hospital stays by 4-12 days. Each year, RVGE incurred $0.54- $53.6 million in direct medical costs and $1.7-$22.4 million in indirect costs in the 16 countries studied. Full serotyping data was available for 8 countries. G1P[8], G2P[4], G9P[8], and G3P[8] were the most prevalent serotypes (cumulative frequency: 57.2%- 98.7%). Serotype distribution in nosocomial RVGE was similar.Conclusions: This review confirms that RVGE is a common disease associated with significant morbidity and costs across Western Europe. A vaccine protecting against multiple serotypes may decrease the epidemiological and cost burden of RVGE in Western Europe. © 2012 Ogilvie et al; licensee BioMed Central Ltd.

Tam J.,Hospital for Sick Children | Beilhartz G.L.,Hospital for Sick Children | Auger A.,Hospital for Sick Children | Gupta P.,Merck And Co. | And 3 more authors.
Chemistry and Biology | Year: 2015

Clostridium difficile causes life-threatening diarrhea through the actions of its homologous toxins TcdA and TcdB on human colonocytes. Therapeutic agents that block toxin-induced damage are urgently needed to prevent the harmful consequences of toxin action that are not addressed with current antibiotic-based treatments. Here, we developed an imaging-based phenotypic screen to identify small molecules that protected human cells from TcdB-induced cell rounding. A series of structurally diverse compounds with antitoxin activity were identified and found to act through one of a small subset of mechanisms, including direct binding and sequestration of TcdB, inhibition of endosomal maturation, and noncompetitive inhibition of the toxin glucosyltransferase activity. Distinct classes of inhibitors were used further to dissect the determinants of the toxin-mediated necrosis phenotype occurring at higher doses of toxin. These findings validate and inform novel targeting strategies for discovering small molecule agents to treat C. difficile infection. © 2015 Elsevier Ltd.

Xu X.,Scripps Research Institute | Shi Y.,Scripps Research Institute | Zhang H.-M.,Florida State University | Zhang H.-M.,Merck And Co. | And 5 more authors.
Nature Communications | Year: 2012

New domains were progressively added to cytoplasmic aminoacyl transfer RNA (tRNA) synthetases during evolution. One example is the UNE-S domain, appended to seryl-tRNA synthetase (SerRS) in species that developed closed circulatory systems. Here we show using solution and crystal structure analyses and in vitro and in vivo functional studies that UNE-S harbours a robust nuclear localization signal (NLS) directing SerRS to the nucleus where it attenuates vascular endothelial growth factor A expression. We also show that SerRS mutants previously linked to vasculature abnormalities either deleted the NLS or have the NLS sequestered in an alternative conformation. A structure-based second-site mutation, designed to release the sequestered NLS, restored normal vasculature. Thus, the essential function of SerRS in vascular development depends on UNE-S. These results are the first to show an essential role for a tRNA synthetase-associated appended domain at the organism level, and suggest that acquisition of UNE-S has a role in the establishment of the closed circulatory systems of vertebrates. © 2012 Macmillan Publishers Limited. All rights reserved.

Liu Q.,University of Washington | Zhang J.,University of Washington | Zerbinatti C.,University of Washington | Zerbinatti C.,Merck And Co. | And 6 more authors.
PLoS Biology | Year: 2011

Obesity is a growing epidemic characterized by excess fat storage in adipocytes. Although lipoprotein receptors play important roles in lipid uptake, their role in controlling food intake and obesity is not known. Here we show that the lipoprotein receptor LRP1 regulates leptin signaling and energy homeostasis. Conditional deletion of the Lrp1 gene in the brain resulted in an obese phenotype characterized by increased food intake, decreased energy consumption, and decreased leptin signaling. LRP1 directly binds to leptin and the leptin receptor complex and is required for leptin receptor phosphorylation and Stat3 activation. We further showed that deletion of the Lrp1 gene specifically in the hypothalamus by Cre lentivirus injection is sufficient to trigger accelerated weight gain. Together, our results demonstrate that the lipoprotein receptor LRP1, which is critical in lipid metabolism, also regulates food intake and energy homeostasis in the adult central nervous system. © 2011 Liu et al.

Wu Y.,University of Virginia | Simons J.,Merck And Co. | Hooson S.,Merck And Co. | Abraham D.,Merck And Co. | Carta G.,University of Virginia
Journal of Chromatography A | Year: 2013

The structural and protein adsorption characteristics of the perfusion chromatography matrix POROS® HS 50 are determined. Transmission electron microscopy shows a broad distribution of pore sizes with 100-500nm through-pores transecting a network of much smaller pores formed by aggregates of microgranules about 100nm in size. Dextran standards, proteins, and virus-like particles (VLPs) show size-exclusion behavior consistent with such a bimodal distribution of pore sizes. For non-binding conditions, the trends in height equivalent to a theoretical plate (HETP) as a function of mobile phase velocity and molecular size are consistent with perfusion suggesting that a fraction of the mobile phase between 0.0005 and 0.0008 flows through the particles. This small fraction provides little or no enhancement of intraparticle mass transfer for relatively small proteins (lysozyme and IgG) even at 1000cm/h, but can contribute substantially to transport for large proteins (thyroglobulin) and VLPs. Intraparticle concentration profiles during transient adsorption are determined by confocal microscopy in batch and flow systems. The profiles are spherically symmetrical indicating a dominance of diffusion for smaller proteins in both batch and flow systems but become highly asymmetrical and skewed in the direction of flow for thyroglobulin at 1000cm/h. Estimates of the convective enhancement of intraparticle transport for these conditions based on the confocal measurements are consistent with estimates of the intraparticle Peclet number and previously published models. Adsorption of VLPs, however, was found to be confined to a thin layer on the outer surface of the particles indicting that bound VLPs block access to the underlying pore network and suggesting that pores larger than those present on the resin studies are needed to take advantage of the effects of perfusion for the adsorption of large VLPs. © 2013 Elsevier B.V.

La Rocca G.,Thomas Jefferson University | Shi B.,Merck And Co. | Sepp-Lorenzino L.,Merck And Co. | Baserga R.,Thomas Jefferson University
Journal of Cellular Physiology | Year: 2011

Micro-RNA-145 (miR145), a tumor suppressor miR, dramatically inhibits growth of cancer cells in culture and plays a significant role in human stem cells differentiation. We have isolated a human genomic sequence of 864 bp comprising the pre-miR and its flanking sequences. The cloned miR145 genomic sequence expresses a mature miR145 in transfected cells. We show here that flanking sequences on either side of the pre-miR sequence can modulate its expression levels. Surprisingly, a highly conserved sequence 3' to the pre-miR plays a crucial role in miR145 expression. © 2010 Wiley-Liss, Inc.

Leiser S.C.,Lundbeck Research United States Inc. | Dunlop J.,Pfizer | Bowlby M.R.,Merck And Co. | Devilbiss D.M.,University of Wisconsin - Madison
Biochemical Pharmacology | Year: 2011

Electroencephalography (EEG) and related methodologies offer the promise of predicting the likelihood that novel therapies and compounds will exhibit clinical efficacy early in preclinical development. These analyses, including quantitative EEG (e.g. brain mapping) and evoked/event-related potentials (EP/ERP), can provide a physiological endpoint that may be used to facilitate drug discovery, optimize lead or candidate compound selection, as well as afford patient stratification and Go/No-Go decisions in clinical trials. Currently, the degree to which these different methodologies hold promise for translatability between preclinical models and the clinic have not been well summarized. To address this need, we review well-established and emerging EEG analytic approaches that are currently being integrated into drug discovery programs throughout preclinical development and clinical research. Furthermore, we present the use of EEG in the drug development process in the context of a number of major central nervous system disorders including Alzheimer's disease, schizophrenia, depression, attention deficit hyperactivity disorder, and pain. Lastly, we discuss the requirements necessary to consider EEG technologies as a biomarker. Many of these analyses show considerable translatability between species and are used to predict clinical efficacy from preclinical data. Nonetheless, the next challenge faced is the selection and validation of EEG endpoints that provide a set of robust and translatable biomarkers bridging preclinical and clinical programs. © 2010 Elsevier Inc.

Wachsberger P.R.,Thomas Jefferson University | Lawrence Y.R.,Thomas Jefferson University | Liu Y.,Thomas Jefferson University | Daroczi B.,Thomas Jefferson University | And 2 more authors.
International Journal of Radiation Oncology Biology Physics | Year: 2012

Purpose: The purpose of this study was to determine the ability of radiation therapy (RT) combined with the tyrosine kinase inhibitors (TKI) vandetanib (antiepidermal growth factor receptor [EGFR] plus antivascular endothelial growth factor receptor [anti-VEGFR]) and cediranib (anti-VEGFR) to inhibit glioblastoma multiforme (GBM) growth. A secondary aim was to investigate how this regimen is modulated by tumor EGFR expression. Methods and Materials: Radiosensitivity was assessed by clonogenic cell survival assay. VEGF secretion was quantified by enzyme-linked immunosorbent assay. GBM (U87MG wild-type EGFR [wtEGFR] and U87MG EGFR-null) xenografts were treated with vandetanib, cediranib, and RT, alone or in combinations. Excised tumor sections were stained for proliferative and survival biomarkers. Results: In vitro, U87MG wtEGFR and U87 EGFR-null cells had similar growth kinetics. Neither TKI affected clonogenic cell survival following RT. However, in vivo, exogenous overexpression of wtEGFR decreased tumor doubling time (T2x) in U87MG xenografts (2.70 vs. 4.41 days for U87MG wtEGFR vs. U87MG vector, respectively). In U87MG EGFR-null cells, TKI combined with radiation was no better than radiation therapy alone. In U87MG wtEGFR, RT in combination with vandetanib (but not with cediranib) significantly increased tumor T2x compared with RT alone (T2x, 10.4 days vs. 4.8 days; p < 0.001). In vivo, growth delay correlated with suppression of pAkt, survivin, and Ki67 expression in tumor samples. The presence of EGFR augmented RT-stimulated VEGF release; this effect was inhibited by vandetanib. Conclusions: EGFR expression promoted tumor growth in vivo but not in vitro, suggesting a microenvironmental effect. GBM xenografts expressing EGFR exhibited greater sensitivity to both cediranib and vandetanib than EGFR-null tumors. Hence EGFR status plays a major role in determining a tumor's in vivo response to radiation combined with TKI, supporting a "personalized" approach to GBM management. Copyright © 2012 Elsevier Inc. Printed in the USA. All rights reserved.

Emmerich J.,Merck And Co. | Emmerich J.,TriMod Therapeutics | Mumm J.B.,Merck And Co. | Mumm J.B.,Targenics Inc. | And 7 more authors.
Cancer Research | Year: 2012

The presence of activated intratumoral T cells correlates clinically with better prognosis in patients with cancer. Although tumor vaccines can increase the number of tumor-specific CD8+ T cells in systemic circulation, they frequently fail to increase the number of active and tumor reactive T cells within the tumor. Here we show that treatment with the pleiotropic cytokine interleukin-10 (IL-10) induces specific activation of tumor-resident CD8 + T cells as well as their intratumoral expansion in several mouse tumor models. We found that inhibition of T-cell trafficking from lymphoid organs did not impair IL-10-induced tumor rejection or the activation of tumor-resident CD8+ T cells. Tumor-resident CD8+ T cells expressed elevated levels of the IL-10 receptor and were directly activated by IL-10, resulting in prominent phosphorylation of STAT3 and STAT1. Although CD4+ T cells, regulatory T cells, NK cells, and dendritic cells have been reported as prominent targets of IL-10 in the tumor microenvironment, we found that expression of the IL-10R was required only on CD8+ T cells to facilitate IL-10-induced tumor rejection as well as in situ expansion and proliferation of tumor-resident CD8 T cells. Together, our findings indicate that IL-10 activates CD8+ T-cell-mediated tumor control and suggest that IL-10 may represent a potential tumor immunotherapy in human patients with cancer. ©2012 AACR.

Guan H.-P.,Merck And Co. | Chen G.,University of Tennessee at Knoxville
Progress in Molecular Biology and Translational Science | Year: 2014

Obesity has become a major concern of public health. A common feature of obesity and related metabolic disorders such as noninsulin-dependent diabetes mellitus is insulin resistance, wherein a given amount of insulin produces less than normal physiological responses. Insulin controls hepatic glucose and fatty acid metabolism, at least in part, via the regulation of gene expression. When the liver is insulin-sensitive, insulin can stimulate the expression of genes for fatty acid synthesis and suppress those for gluconeogenesis. When the liver becomes insulin-resistant, the insulin-mediated suppression of gluconeogenic gene expression is lost, whereas the induction of fatty acid synthetic gene expression remains intact. In the past two decades, the mechanisms of insulin-regulated hepatic gene expression have been studied extensively and many components of insulin signal transduction pathways have been identified. Factors that alter these pathways, and the insulin-regulated hepatic gene expression, have been revealed and the underlying mechanisms have been proposed. This chapter summarizes the recent grogresses in our understanding of the effects of dietary factors, drugs, bioactive compounds, hormones, and cytokines on insulin-regulated hepatic gene expression. Given the large amount of information and progresses regarding the roles of insulin, this chapter focuses on findings in the liver and hepatocytes and not those described for other tissues and cells. Typical insulin-regulated hepatic genes, such as insulin-induced glucokinase and sterol regulatory element-binding protein-1c and insulin-suppressed cytosolic phosphoenolpyruvate carboxyl kinase and insulin-like growth factor-binding protein 1, are used as examples to discuss the mechanisms such as insulin regulatory element-mediated transcriptional regulation. We also propose the potential mechanisms by which these factors affect insulin-regulated hepatic gene expression and discuss potential future directions of the area of research. © 2014 Elsevier Inc.

Modi A.,Merck And Co. | Siris E.S.,Columbia University | Yang X.,Temple University | Fan C.-P.S.,Asclepius JT | Sajjan S.,Merck And Co.
Journal of Managed Care Pharmacy | Year: 2015

BACKGROUND: A large proportion of patients do not persist with osteoporosis (OP) therapy. Gastrointestinal (GI) events (e.g., gastroesophageal reflux disease and nausea/vomiting) are common among OP patients receiving OP therapy and may impact persistence with treatment. Objective: To examine the association of GI events and persistence with OP therapy. METHODS: Using a large U.S. administrative claims database, we studied women aged ≥ 55 years who received oral bisphosphonate (BIS) as their first OP therapy from 2002-2009. The index date was the first pharmacy claim date recorded for oral BIS therapy; the baseline period was 12 months pre-index, and follow-up was 12 months post-index. Patients were considered persistent with therapy if they had continuous refills of the index drug class without additional drug therapy from a different drug class from the index date until the end of the follow-up period with no gaps in supply greater than 45 days. Discontinuation was defined as the first gap greater than 45 days during which there was no evidence of refills of OP medication. The association between post-treatment GI events and the risk of discontinuation or switching was modeled with Cox regression stratified by presence of baseline GI events and adjusted for baseline clinical and demographic characteristics. RESULTS: Of the 75,593 women who met eligibility criteria, 59.9% discontinued BIS; 39.3% were persistent; and 0.5% switched to non-BIS. GI events were diagnosed in 20,073 patients (26.6%) during baseline and in 21,142 (28.0%) in the post-treatment period (12-month follow-up postindex). Patients with post-treatment GI diagnosis were 35.6% more likely to discontinue or switch treatment (HR = 1.356, 95% CI = 1.318-1.396) during the 12-month follow-up compared with those without post-treatment GI diagnosis. GI events that occurred closer to treatment discontinuation or switching were associated with a greater risk of discontinuation or switching: 37.9% (HR = 1.379, 95% CI = 1.338-1.421) for GI events within 6 months of discontinuation or switching and 45.6% (HR = 1.456, 95% CI = 1.408-1.505) for GI events within 3 months of discontinuation or switching. CONCLUSIONS: Among women aged 55 years or older in a U.S. managed care population, post-treatment GI events were associated with a higher risk of discontinuation of oral BIS or switching to non-BIS. © 2015, Academy of Managed Care Pharmacy.

Calix J.J.,University of Alabama at Birmingham | Nahm M.H.,University of Alabama at Birmingham | Zartler E.R.,Merck And Co.
Journal of Bacteriology | Year: 2011

Despite the emerging impact of serogroup 11 serotypes in Streptococcus pneumoniae epidemiology, the structures of serogroup 11 capsule types have not been fully elucidated, particularly the locations of O-acetyl substitutions. Here, we report the complete structures of the serotype 11B, 11C, and 11F polysaccharides and a revision to the serotype 11A capsular polysaccharide using nuclear magnetic resonance (NMR). All structures shared a linear, tetrasaccharide backbone with a pendant phosphopolyalcohol. Three of four saccharides are conserved in all serotypes. The individual serotype capsules differed in the identity of one saccharide, the pendant phosphopolyalcohol, and the O-acetylation pattern. Though the assigned locations of O-acetate substitutions in this study differed from those of previous reports, our findings were corroborated with strong correlations to serology and genetics. We examined the binding of serotyping sera to serogroup 11 polysaccharides by using flow cytometry and an inhibition-type enzyme-linked immunosorbent assay (ELISA) and found that de-O-acetylation of capsular polysaccharides by mild hydrolysis decreases its immunoreactivity, supporting the crucial role of O-acetylation in the antigenicity of these polysaccharides. Due to strong correlations between polysaccharide structures and capsule biosynthesis genes, we were able to assign target substrates for the O-acetyltransferases encoded by wcwC, wcwR, wcwT, and wcjE. We identified antigenic determinants for serogroup 11 serotyping sera and highlight the idea that conventional serotyping methods are not capable of recognizing all putative variants of S. pneumoniae serogroup 11. © 2011, American Society for Microbiology.

Neale S.A.,University College London | Neale S.A.,Neurexpert Ltd. | Copeland C.S.,University College London | Uebele V.N.,Merck And Co. | And 2 more authors.
Neuropsychopharmacology | Year: 2013

Xanthurenic acid (XA), an endogenous kynurenine, is a known vesicular glutamate transport (VGLUT) inhibitor and has also been proposed as an mGlu2/3 receptor agonist. Changes in these systems have been implicated in the pathophysiology of schizophrenia and other psychiatric disorders; however, little is known of how XA affects synaptic transmission. We therefore investigated the effects of XA on synaptic transmission at two hippocampal glutamatergic pathways and evaluated the ability of XA to bind to mGlu2/3 receptors. Field excitatory postsynaptic potentials (fEPSPs) were recorded from either the dentate gyrus (DG) or CA1 region of mouse hippocampal slices in vitro. Addition of XA to the bathing medium (1-10 mM) resulted in a dose-related reduction of fEPSP amplitudes (up to 52% reduction) in both hippocampal regions. In the DG, the VGLUT inhibitors Congo Red and Rose Bengal, and the mGlu2/3 agonist LY354740, also reduced fEPSPs (up to 80% reduction). The mGlu2/3 antagonist LY341495 reversed the LY354740 effect, but not the XA effect. LY354740, but not XA, also reduced DG paired-pulse depression. XA had no effect on specific binding of 1 nM [ 3 H]LY341495 to membranes with human mGlu2 receptors. We conclude that XA can modulate synaptic transmission via a mechanism that may involve VGLUT inhibition rather than activation of mGlu2/3 receptors. This could be important in the pathophysiology of nervous system disorders including schizophrenia and might represent a target for developing novel pharmacological therapies. © 2013 American College of Neuropsychopharmacology.

Hu-Lieskovan S.,University of California at Los Angeles | Mok S.,University of California at Los Angeles | Homet Moreno B.,University of California at Los Angeles | Homet Moreno B.,Carlos III Health Institute | And 9 more authors.
Science Translational Medicine | Year: 2015

Combining immunotherapy and BRAF targeted therapy may result in improved antitumor activity with the high response rates of targeted therapy and the durability of responses with immunotherapy. However, the first clinical trial testing the combination of the BRAF inhibitor vemurafenib and the CTLA4 antibody ipilimumab was terminated early because of substantial liver toxicities. MEK [MAPK (mitogen-activated protein kinase) kinase] inhibitors can potentiate the MAPK inhibition in BRAF mutant cells while potentially alleviating the unwanted paradoxical MAPK activation in BRAF wild-type cells that lead to side effects when using BRAF inhibitors alone. However, there is the concern of MEK inhibitors being detrimental to T cell functionality. Using a mouse model of syngeneic BRAFV600E-driven melanoma, SM1, we tested whether addition of the MEK inhibitor trametinib would enhance the antitumor activity of combined immunotherapy with the BRAF inhibitor dabrafenib. Combination of dabrafenib and trametinib with pmel-1 adoptive cell transfer (ACT) showed complete tumor regression, increased T cell infiltration into tumors, and improved in vivo cytotoxicity. Single-agent dabrafenib increased tumor-associated macrophages and T regulatory cells (Tregs) in tumors, which decreased with the addition of trametinib. The triple combination therapy resulted in increased melanosomal antigen and major histocompatibility complex (MHC) expression and global immune-related gene up-regulation. Given the up-regulation of PD-L1 seen with dabrafenib and/or trametinib combined with antigen-specific ACT, we tested the combination of dabrafenib, trametinib, and anti-PD1 therapy in SM1 tumors, and observed superior antitumor effect. Our findings support the testing of triple combination therapy of BRAF and MEK inhibitors with immunotherapy in patients with BRAFV600E mutant metastatic melanoma. © 2015, American Association for the Advancement of Science. All rights reserved.

Stuginski-Barbosa J.,University of Sao Paulo | Dach F.,University of Sao Paulo | Bigal M.,Merck And Co. | Speciali J.G.,University of Sao Paulo
Headache | Year: 2012

Background. Migraine is comorbid to depression and widespread chronic pain (WCP), but the influence of these conditions on the health-related quality of life (HRQoL) of individuals with episodic (EM) and chronic migraine (CM) is poorly understood. Objective. To assess the prevalence of depressive symptoms and WCP in individuals with EM and CM, as well as to estimate the joint impact of these conditions on the HRQoL of these individuals. Methods. All women aged 18 to 65 years with a first diagnosis of EM or CM from September of 2006 to September of 2008 seen in an outpatient headache service were invited to participate. They were asked to attend a separate appointment in the service, and to bring another woman of similar age that also agreed to participate. Depressive symptoms were assessed using the Beck Depression Inventory. Questions about WCP followed the protocol of the American College of Rheumatology. HRQoL was assessed using the Short-Form 36 (SF-36). Multivariate analysis modeled HRQoL as a function of headache status, depressive symptoms, and pain, using quantile regression. Results. Sample consisted of 179 women, 53 in the EM group, 37 in the CM group and 89 in control group. Groups did not differ by demographics. Mean scores of SF-36 were 53.6 (standard deviation [SD] = 23.5) for EM, 44.2 (SD = 18.5) for CM and 61.8 (SD = 21.5) for controls. In multivariate analysis, SF-36 scores were predicted by a CM status (P =.02; -10.05 [95% CI -18.52; -1.58]) and by a Beck Depression Inventory score (P <.01; -1.27 [95% CI -1.55; -0.99]). The influence of WCP in the SF-36 scores approached significance (P =.08; -0.78 [95% CI -1.64; 0.88]). Age did not contribute to the model. Conclusion. Women with migraine are at an increased chance of WCP, and the chance increases as a function of headache frequency. Both depressive symptoms and CM independently predict HRQoL status. © 2012 American Headache Society.

Prenner B.M.,Allergy Associates Medical Group | Lanier B.Q.,University of North Texas | Bernstein D.I.,University of Cincinnati | Shekar T.,Merck And Co. | Teper A.,Merck And Co.
Journal of Allergy and Clinical Immunology | Year: 2010

Background: Mometasone furoate nasal spray (MFNS), a potent intranasal corticosteroid with proved efficacy in relieving nasal allergic rhinitis symptoms, has demonstrated effectiveness in improving ocular symptoms associated with seasonal allergic rhinitis (SAR) in retrospective analyses. Objective: We sought to evaluate prospectively the efficacy of MFNS in reducing total ocular symptom scores (TOSSs) and individual ocular symptoms in subjects with SAR. Methods: Subjects 12 years or older (n = 429) with moderate-to-severe baseline symptoms were randomized to MFNS, 200 μg once daily, or placebo in this 15-day, double-blind, parallel-group study. Subjects evaluated morning instantaneous TOSSs and daily reflective TOSSs, total nasal symptom scores (TNSSs; both instantaneous TNSSs and reflective TNSSs, respectively), and individual ocular and nasal symptoms. Mean changes from baseline averaged over days 2 to 15 (instantaneous) and days 1 to 15 (reflective) were calculated. Quality of life was assessed by using the Rhinoconjunctivitis Quality of Life Questionnaire. Results: MFNS treatment yielded significant reductions from baseline versus placebo in instantaneous TOSSs (-0.34, P = .026, coprimary end point), instantaneous TNSSs (-0.88, P < .001, coprimary end point), reflective TOSSs (-0.44, P = .005), and reflective TNSSs (-1.06, P < .001). Significant decreases in all individual reflective ocular symptoms and instantaneous eye itching/burning and eye watering/tearing were observed for MFNS versus placebo (P < .05). Numeric improvements in instantaneous eye redness were seen but did not reach statistical significance. Improvements in Rhinoconjunctivitis Quality of Life Questionnaire total scores and individual symptom domains were achieved with MFNS treatment versus placebo (P < .001). MFNS was well tolerated. Conclusion: This prospective study demonstrates that MFNS significantly reduces ocular symptoms in subjects with SAR. © 2010 American Academy of Allergy, Asthma & Immunology.

Panaccione R.,University of Calgary | Ghosh S.,University of Calgary | Middleton S.,University of Cambridge | Marquez J.R.,Clinica Las Americas | And 7 more authors.
Gastroenterology | Year: 2014

Background & Aims The comparative efficacy and safety of infliximab and azathioprine therapy alone or in combination for ulcerative colitis (UC) have not been evaluated previously. Methods This randomized, double-blind trial evaluated the efficacy and safety of 16 weeks of treatment with infliximab monotherapy, azathioprine monotherapy, or the 2 drugs combined in tumor necrosis factor-α antagonist-naive adults with moderate to severe UC. Patients were assigned randomly to receive intravenous infusions of infliximab 5 mg/kg at weeks 0, 2, 6, and 14 plus daily oral placebo capsules; oral azathioprine 2.5 mg/kg daily plus placebo infusions on the infliximab schedule; or combination therapy with the 2 drugs. Corticosteroid-free clinical remission (primary end point, week 16) was evaluated at weeks 8 and 16. The study was terminated before the enrollment target was reached. Results A total of 239 patients were included in efficacy analyses. Baseline characteristics were similar between treatment groups. Corticosteroid-free remission at week 16 was achieved by 39.7% (31 of 78) of patients receiving infliximab/azathioprine, compared with 22.1% (17 of 77) receiving infliximab alone (P =.017) and 23.7% (18 of 76) receiving azathioprine alone (P =.032). Mucosal healing at week 16 occurred in 62.8% (49 of 78) of patients receiving infliximab/azathioprine, compared with 54.6% (42 of 77) receiving infliximab (P =.295) and 36.8% (28 of 76) receiving azathioprine (P =.001). Serious infections occurred in 2 patients (1 patient receiving infliximab, and 1 patient receiving azathioprine). Conclusions Anti-tumor necrosis factor-α-naive patients with moderate to severe UC treated with infliximab plus azathioprine were more likely to achieve corticosteroid-free remission at 16 weeks than those receiving either monotherapy. Combination therapy led to significantly better mucosal healing than azathioprine monotherapy. ClinicalTrials.gov number, NCT00537316. © 2014 by the AGA Institute.

Ahern P.P.,University of Oxford | Schiering C.,University of Oxford | Schiering C.,John Radcliffe Hospital | Buonocore S.,University of Oxford | And 5 more authors.
Immunity | Year: 2010

Mutations in the IL23R gene are linked to inflammatory bowel disease susceptibility. Experimental models have shown that interleukin-23 (IL-23) orchestrates innate and T cell-dependent colitis; however, the cell populations it acts on to induce intestinal immune pathology are unknown. Here, using Il23r-/- T cells, we demonstrated that T cell reactivity to IL-23 was critical for development of intestinal pathology, but not for systemic inflammation. Through direct signaling into T cells, IL-23 drove intestinal T cell proliferation, promoted intestinal Th17 cell accumulation, and enhanced the emergence of an IL-17A+IFN-γ+ population of T cells. Furthermore, IL-23R signaling in intestinal T cells suppressed the differentiation of Foxp3+ cells and T cell IL-10 production. Although Il23r-/- T cells displayed unimpaired Th1 cell differentiation, these cells showed impaired proliferation and failed to accumulate in the intestine. Together, these results highlight the multiple functions of IL-23 signaling in T cells that contribute to its colitogenic activity. © 2010 Elsevier Inc.

Patel H.,TransCanada | Wagner M.,TransCanada | Singhal P.,Merck And Co. | Kothari S.,Merck And Co.
BMC Infectious Diseases | Year: 2013

Background: Anogenital warts (AGWs) are a common, highly infectious disease caused by the human papillomavirus (HPV), whose high recurrence rates contribute to direct medical costs, productivity loss and increased psychosocial impact. Because of the lack of a systematic review of the epidemiology of AGWs in the literature, this study reviewed the published medical literature on the incidence and prevalence of AGWs. Methods: A comprehensive literature search was performed on the worldwide incidence and prevalence of AGWs between 2001 and 2012 using the PubMed and EMBASE databases. An additional screening of abstracts from relevant sexual health and infectious disease conferences from 2009 to 2011 was also conducted. Only original studies with general adult populations (i.e., at least including ages 20 through 40 years) were included. Results: The overall (females and males combined) reported annual incidence of any AGWs (including new and recurrent) ranged from 160 to 289 per 100,000, with a median of 194.5 per 100,000. New AGW incidence rates among males ranged from 103 to 168 per 100,000, with a median of 137 per 100,000 and among females from 76 to 191 per 100,000, with a median of 120.5 per 100,000 per annum. The reported incidence of recurrent AGWs was as high as 110 per 100,000 among females and 163 per 100,000 among males. Incidence peaked before 24 years of age in females and between 25 and 29 years of age among males. The overall prevalence of AGWs based on retrospective administrative databases or medical chart reviews or prospectively collected physician reports ranged from 0.13% to 0.56%, whereas it ranged from 0.2% to 5.1% based on genital examinations. Conclusions: The literature suggests that AGWs are widespread and the prevalence depends on study methodology as suggested by higher rates reported from routine genital examinations versus those from treatment records. However, there remains a need for more population-based studies from certain regions including Africa, Latin America and Southern Asia to further elucidate the global epidemiology of this disease. © 2013 Patel et al.; licensee BioMed Central Ltd.

Khoury H.,TransCanada | Ogilvie I.,TransCanada | El Khoury A.C.,Merck And Co. | Duan Y.,Lehigh University | Goetghebeur M.M.,TransCanada
BMC Infectious Diseases | Year: 2011

Background: Rotavirus gastroenteritis (RVGE) is the most common cause of severe childhood diarrhea worldwide. Objectives were to estimate the burden of RVGE among children less than five years old in the Middle East (Bahrain, Iran, Iraq, Israel, Jordan, Kuwait, Oman, Qatar, Saudi Arabia, Syria, UAE, Yemen), North Africa (Algeria, Egypt, Libya, Morocco, Tunisia) and Turkey.Methods: A comprehensive literature search was conducted in major databases on the epidemiology and burden of rotavirus among children less than five years old between 1999 and 2009. Data from each country was extracted and compared.Results: The search identified 43 studies. RVGE was identified in 16-61% of all cases of acute gastroenteritis, with a peak in the winter. RVGE-related hospitalization rates ranged from 14% to 45%, compared to 14%-28% for non-RVGE. Annually, RVGE caused up to 112 fatalities per 100,000 in certain countries in the region. Hospitalization costs ranged from $1.8 to $4.6 million annually, depending on the country. The most recent literature available showed that G1P[8] was the most prevalent genotype combination in 8 countries (range 23%-56%). G2P[4] was most prevalent in 4 countries (26%-48%). G9P[8] and G4P[8] were also frequently detected.Conclusions: RVGE is a common disease associated with significant morbidity, mortality, and economic burden. Given the variety and diverse rotavirus types in the region, use of a vaccine with broad and consistent serotype coverage would be important to help decrease the burden of RVGE in the Middle East and North Africa. © 2011 Khoury et al; licensee BioMed Central Ltd.

Levy M.J.,Shady Grove Fertility | Ledger W.,University of New South Wales | Kolibianakis E.M.,Aristotle University of Thessaloniki | IJzerman-Boon P.C.,Oss | Gordon K.,Merck And Co.
Reproductive BioMedicine Online | Year: 2013

A retrospective analysis of a large, randomized clinical trial (Engage) assessed whether adjusting the start day of ovarian stimulation and/or day of human chorionic gonadotrophin (HCG) trigger could minimize oocyte retrieval during weekends without adverse effects on clinical outcome. Patients received recombinant FSH/gonadotrophin-releasing hormone (GnRH) antagonist regimens, with stimulation starting on day 2 or 3 of menses. HCG was administered when at least three follicles of 17 mm were present on ultrasound scan or 1 day later. The frequency distribution of the day of reaching the HCG criterion relative to stimulation initiation was analysed to determine the optimal stimulation start day (cycle day 2 or 3) depending on the weekday at which menses started, to minimize weekend retrieval. The number of oocytes retrieved and pregnancy rates were not affected by start day and/or delay in HCG administration in regularly ovulating women aged 18-36 years with bodyweight 60-90 kg, body mass index 18-32 kg/m2 and menstrual cycle length 24-35 days. In recombinant FSH/GnRH antagonist regimens, it appears possible to minimize weekend oocyte retrieval by selecting the cycle day to initiate stimulation, day 2 when menses starts Friday-Tuesday, otherwise day 3 and if necessary in combination with a 1-day HCG delay. © 2012, Reproductive Healthcare Ltd.

Zhao M.,University of East Anglia | Barker S.A.,University of East Anglia | Belton P.S.,University of East Anglia | McGregor C.,Merck And Co. | Craig D.Q.M.,University of East Anglia
European Journal of Pharmaceutics and Biopharmaceutics | Year: 2012

The aim of the study was to prepare molecular dispersions of a physically highly unstable amorphous drug, paracetamol (acetaminophen with a Tg of ca. 25 °C) via co-spray drying with a variety of polymers. Solid dispersions at a range of drug loadings (10-90%w/w) using hydroxypropyl methylcellulose/acetate succinate (HPMC/HPMC AS), polyvinylpyrrolidone (PVP) and copovidone were produced and characterised by modulated temperature differential scanning calorimetry (MTDSC), thermogravimetric analysis (TGA), X-ray powder diffraction (XRPD), Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). PVP-based polymers showed a greater tendency than the HPMC-based group to generate temperature-stable dispersions. In particular, copovidone (Plasdone® S-630) was found to be the most effective of the polymers studied and could formulate molecular dispersions at drug loadings up to and including 40%w/w. However, no evidence for direct drug-polymer interactions was found for such systems as a possible stabilising mechanism. The expected relationship of a higher Tg of the polymer leading to greater stabilisation was not observed, while there was an inverse relationship between viscosity grade and amorphous phase generation. The study has therefore shown that temperature-stable amorphous dispersions of a low Tg drug may be prepared by co-spray drying, particularly using PVP-based polymers. © 2012 Elsevier B.V. All rights reserved.

Sherlock J.P.,University of Birmingham | Sherlock J.P.,University of Oxford | Buckley C.D.,University of Birmingham | Cua D.J.,Merck And Co.
Molecular Immunology | Year: 2014

The spondyloarthropathies represent highly enigmatic conditions and although their clinical features, anatomical distribution of disease and genetic predisposing factors have been known for some time, a unified concept of the basic pathobiology underlying these illnesses has remained undefined. Recently progress has been made because numerous independent studies have converged upon IL-23 as a central cytokine in spondyloarthropathy and the mechanism and sites of action of this cytokine have now become much clearer. These findings enable the rational design of therapeutic strategies which it is hoped will profoundly modify the progression of these diseases. We will review the anatomical sites affected and the evidence for the importance of IL-23 in these conditions, before drawing these lines of investigation together to propose a model for the unified understanding of spondyloarthropathy. © 2013 Elsevier Ltd.

Tang W.,Merck And Co. | Lu A.Y.H.,Rutgers University
Drug Metabolism Reviews | Year: 2010

Retrospective studies indicate that many drugs that cause clinical adverse reactions, such as hepatotoxicity, undergo metabolic bioactivation, resulting in the formation of electrophilic intermediates capable of covalently modifying biological macromolecules. A logical extension of these findings is a working hypothesis that compounds with reduced levels of bioactivation should be inherently safer drug molecules and thus have a greater likelihood of success in drug development. Whereas some research-based pharmaceutical companies have adopted a strategy of addressing metabolic bioactivation early in drug discovery, much skepticism remains on whether such an approach would enable the industry to reach the desired objectives. The debate is centered on the question of whether there is a quantitative correlation between bioactivation and the severity of drug-treatmentrelated toxicity, and whether covalent protein modification represents only one of several possible mechanisms underlying observed tissue injury. This communication is intended to briefly review the current understanding of drug-induced hepatotoxicity and to discuss the controversy and future directions with respect to the effort of minimizing the probability of clinical adverse reactions. © 2010 Informa UK Ltd.

Staab J.F.,Johns Hopkins University | Kahn J.N.,Merck And Co. | Kahn J.N.,Rutgers University | Marr K.A.,Johns Hopkins University
Antimicrobial Agents and Chemotherapy | Year: 2010

The recently described species Aspergillus lentulus exhibits differential and reduced susceptibilities to echinocandins and other antifungal drugs in vitro. A. lentulus isolates overall are less susceptible to caspofungin, although they maintain susceptibility to anidulafungin and micafungin. Mutations or polymorphisms in fks, the gene encoding the catalytic subunit of β-1,3-glucan synthase, are known to confer decreased susceptibility to echinocandins in Candida spp. and Aspergillus fumigatus. The analysis of the A. lentulus fks sequence did not reveal a polymorphism at any of the known hot-spot regions of the gene. Caspofungin and micafungin kinetic inhibition profiles of the A. lentulus glucan synthase were comparable to those from susceptible A. fumigatus enzymes. Although the basal cell wall chitin levels in A. lentulus averaged 60% of those in A. fumigatus, echinocandin treatment promoted the increase of cell wall chitin in both organisms, indicating that A. lentulus displays a compensatory chitin response similar to that of A. fumigatus. The data suggest that differential echinocandin susceptibilities in A. lentulus are independent of the echinocandin target, Fksp, and they emphasize the potential that the drugs' capacity to inhibit the target enzyme is unequal at the cellular level. Copyright © 2010, American Society for Microbiology. All Rights Reserved.

Mandl J.N.,U.S. National Institutes of Health | Mandl J.N.,McGill University | Ahmed R.,Emory University | Barreiro L.B.,University of Montréal | And 4 more authors.
Cell | Year: 2015

Zoonotic viruses, such as HIV, Ebola virus, coronaviruses, influenza A viruses, hantaviruses, or henipaviruses, can result in profound pathology in humans. In contrast, populations of the reservoir hosts of zoonotic pathogens often appear to tolerate these infections with little evidence of disease. Why are viruses more dangerous in one species than another? Immunological studies investigating quantitative and qualitative differences in the host-virus equilibrium in animal reservoirs will be key to answering this question, informing new approaches for treating and preventing zoonotic diseases. Integrating an understanding of host immune responses with epidemiological, ecological, and evolutionary insights into viral emergence will shed light on mechanisms that minimize fitness costs associated with viral infection, facilitate transmission to other hosts, and underlie the association of specific reservoir hosts with multiple emerging viruses. Reservoir host studies provide a rich opportunity for elucidating fundamental immunological processes and their underlying genetic basis, in the context of distinct physiological and metabolic constraints that contribute to host resistance and disease tolerance. © 2015 Elsevier Inc. All rights reserved.

Perez-Trujillo M.,Autonomous University of Barcelona | Castanar L.,Autonomous University of Barcelona | Monteagudo E.,Autonomous University of Barcelona | Kuhn L.T.,European Neuroscience Institute Gottingen ENI G | And 4 more authors.
Chemical Communications | Year: 2014

NMR enantiodifferentiation studies are greatly improved by the simultaneous determination of 1H and 13C chemical shift differences through the analysis of highly resolved cross-peaks in spectral aliased pure shift (SAPS) HSQC spectra. This journal is © the Partner Organisations 2014.

Hirsch E.B.,University of Houston | Hirsch E.B.,Northeastern University | Ledesma K.R.,University of Houston | Chang K.-T.,University of Houston | And 3 more authors.
Antimicrobial Agents and Chemotherapy | Year: 2012

Carbapenem-resistant bacteria represent a significant treatment challenge due to the lack of active antimicrobials available. MK-7655 is a novel β-lactamase inhibitor under clinical development. We investigated the combined killing activity of imipenem and MK-7655 against four imipenem-resistant bacterial strains, using a mathematical model previously evaluated in our laboratory. Time-kill studies (TKS) were conducted with imipenem and MK-7655 against a KPC-2-producing Klebsiella pneumoniae isolate (KP6339) as well as 3 Pseudomonas aeruginosa isolates (PA24226, PA24227, and PA24228) with OprD porin deletions and overexpression of AmpC. TKS were performed using 25 clinically achievable concentration combinations in a 5-by-5 array. Bacterial burden at 24 h was determined in triplicate by quantitative culture and mathematically modeled using a three-dimensional response surface. Mathematical model assessments were evaluated experimentally using clinically relevant dosing regimens of imipenem, with or without MK-7655, in a hollow-fiber infection model (HFIM). The combination of imipenem and MK-7655 was synergistic for all strains. Interaction indices were as follows: for KP6339, 0.50 (95% confidence interval [CI], 0.42 to 0.58); for PA24226, 0.60 (95% CI, 0.58 to 0.62); for PA24227, 0.70 (95% CI, 0.66 to 0.74); and for PA24228, 0.55 (95% CI, 0.49 to 0.61). In the HFIM, imipenem plus MK-7655 considerably reduced the bacterial burden at 24 h, while failure with imipenem alone was seen against all isolates. Sustained suppression of bacterial growth at 72 h was achieved with simulated doses of 500 mg imipenem plus 500 mg MK-7655 in 2 (KP6339 and PA24227) strains, and it was achieved in an additional strain (PA24228) when the imipenem dose was increased to 1,000 mg. Additional studies are being conducted to determine the optimal dose and combinations to be used in clinical investigations. Copyright © 2012, American Society for Microbiology. All Rights Reserved.

Quinlan M.,Novartis | Stroup W.,University of Nebraska - Lincoln | Schwenke J.,Applied Research Consultants | Christopher D.,Merck And Co.
Journal of Biopharmaceutical Statistics | Year: 2013

The goal of shelf life estimation is to determine the storage time during which the entire product meets specification with acceptably high probability. The estimated shelf life should be "applicable to all future batches" (ICH Q1E, International Conference on Harmonization, 2003b). There is compelling evidence of issues with the International Conference on Harmonization (ICH) guidelines for shelf life estimation. Issues include fixed batch effects, poolability tests, and confidence intervals for the mean. Two conclusions from evaluating the ICH procedure are that batch effects should be random and that focus should be on a quantile. A procedure is needed that combines random batches with the ICH objective of estimating the minimum batch shelf life. © Taylor & Francis Group, LLC.

Nalls M.A.,U.S. National Institute on Aging | Escott-Price V.,University of Cardiff | Williams N.M.,University of Cardiff | Lubbe S.,University College London | And 3 more authors.
Movement Disorders | Year: 2015

Background: Recent genomewide association study meta-analyses have identified 28 loci associated with risk of Parkinson's disease (PD). We sought to investigate whether these genetic risk factors are associated with PD age at onset. Methods: Genetic risk scores from these loci were calculated for 6,249 cases. Linear regression tested associations between cumulative genetic risk and PD age at onset. Results: Increasing genetic risk scores were associated with earlier age at onset (beta=-0.10, P=2.92 × 10-8, adjusted r2=0.27). Single standard deviation increase in genetic risk score is associated with 37.44 d earlier age at onset. Highest genetic risk was found at 31 to 60 y, onset slightly below average age at onset (AAO). Conclusions: Common genetic risk factors have a small but consistent association with AAO in PD. © 2015 International Parkinson and Movement Disorder Society.

Bergman G.J.D.,Mapi Values | Fan T.,Merck And Co. | McFetridge J.T.,Lehigh University | Sen S.S.,Merck And Co.
Current Medical Research and Opinion | Year: 2010

Background: The efficacy of vitamin D3 in preventing fractures and falls has been explored in a number of clinical trials. However, recent evidence revealed new questions about the adequate doses of vitamin D3 supplementation and its efficacy in fracture prevention independent of calcium supplements for various types of fractures. Objective: To conduct a meta-analysis to estimate the effectiveness of 800 IU daily vitamin D3 supplementation for increasing bone mineral density (BMD) and preventing fractures in postmenopausal women. Methods: Medline and EMBASE were searched for controlled trials comparing the effectiveness of cholecalciferol (vitamin D3) against placebo with or without background calcium supplementation in the treatment of postmenopausal women. Results: Eight controlled trials evaluating the effect of vitamin D3 supplementation with or without calcium were assessed. Of 12658 women included in a Bayesian meta-analysis, 6089 received vitamin D3 (with or without calcium) and 6569 received placebo (with or without calcium). Compared to placebo, vitamin D3 with calcium supplementation showed beneficial effects on the incidence of non-vertebral (odds ratio [OR] 0.77, 95 credibility limit [CL] 0.60.93) and hip (OR 0.70, 95 CL 0.530.90) fractures, while the effects on non-vertebral-non-hip fractures (OR 0.84, 95 CL 0.671.04) % point increase) were associated with more uncertainty. Vitamin D3 supplementation showed a 70% probability of being a better treatment than placebo for the prevention of non-vertebral fractures, hip fractures, and non-vertebral, non-hip fractures. Compared to calcium supplementation, vitamin D3 plus calcium reduced non-vertebral fractures (OR 0.68, 95 CL 0.431.01) and non-vertebral, non-hip fractures (OR 0.64, 95 CL 0.380.99), but did not reduce hip fractures (OR 1.03, 95 CL 0.392.25). Key limitations to this analysis include a small number of studies and heterogeneity in the study populations. Conclusions: This meta-analysis supports the use of vitamin D3 of 800 IU daily to reduce the incidence of osteoporotic non-vertebral, hip, and non-vertebral-non-hip fractures in elderly women. Vitamin D3 with calcium appears to achieve benefits above those attained with calcium supplementation alone for non-vertebral and non-vertebral-non-hip fractures. © 2010 Informa UK Ltd All rights reserved.

McFarland K.N.,University of Florida | Huizenga M.N.,Massachusetts General Hospital | Darnell S.B.,Massachusetts General Hospital | Sangrey G.R.,Massachusetts General Hospital | And 4 more authors.
Human Molecular Genetics | Year: 2014

Transcriptional dysregulation has been proposed to play amajor role in the pathology of Huntington's disease (HD). However, the mechanisms that cause selective downregulation of target genes remain unknown. Previous studies have shown that mutant huntingtin (Htt) protein interacts with a number of transcription factors thereby altering transcription. Here we report that Htt directly interacts with methyl-CpG binding protein 2 (MeCP2) in mouse and cellular models of HD using complimentary biochemical and Fluorescent Lifetime Imaging to measure Fö rster Resonance Energy Transfer approaches. Htt-MeCP2 interactions are enhanced in the presence of the expanded polyglutamine (polyQ) tract and are stronger in the nucleus compared with the cytoplasm. Furthermore, we find increased binding of MeCP2 to the promoter of brain-derived neurotrophic factor (BDNF), a gene that is downregulated in HD, in the presence of mutant Htt. Finally, decreasing MeCP2 levels in mutant Htt-expressing cells using siRNA increases BDNF levels, suggesting that MeCP2 downregulates BDNF expression in HD. Taken together, these findings suggest that aberrant interactions between Htt and MeCP2 contribute to transcriptional dysregulation in HD. © The Author 2013. Published by Oxford University Press. All rights reserved.

Jarajapu Y.P.R.,University of Florida | Bhatwadekar A.D.,University of Florida | Caballero S.,University of Florida | Hazra S.,University of Florida | And 8 more authors.
Diabetes | Year: 2013

We tested the hypothesis that activation of the protective arm of the renin angiotensin system, the angiotensin-converting enzyme 2 (ACE2)/angiotensin-(1- 7) [Ang-(1-7)]/Mas receptor axis, corrects the vasoreparative dysfunction typically seen in the CD34+ cells isolated from diabetic individuals. Peripheral blood CD34+ cells from patients with diabetes were compared with those of nondiabetic controls. Ang-(1-7) restored impaired migration and nitric oxide bioavailability/cGMP in response to stromal cell-derived factor and resulted in a decrease in NADPH oxidase activity. The survival and proliferation of CD34+ cells from diabetic individuals were enhanced by Ang-(1-7) in a Mas/ phosphatidylinositol 3-kinase (PI3K)/Akt-dependent manner. ACE2 expression was lower, and ACE2 activators xanthenone and diminazine aceturate were less effective in inducing the migration in cells from patients with diabetes compared with controls. Ang-(1-7) overexpression by lentiviral gene modification restored both the in vitro vasoreparative functions of diabetic cells and the in vivo homing efficiency to areas of ischemia. A cohort of patients who remained free of microvascular complications despite having a history of longstanding inadequate glycemic control had higher expression of ACE2/Mas mRNA than patients with diabetes with microvascular complications matched for age, sex, and glycemic control. Thus, ACE2/Ang-(1-7)\Mas pathway activation corrects existing diabetes-induced CD34+ cell dysfunction and also confers protection from development of this dysfunction. © 2013 by the American Diabetes Association.

Staprans S.I.,Merck Vaccines | Feinberg M.B.,Merck Vaccines | Shiver J.W.,Merck And Co. | Casimiro D.R.,Merck And Co.
Current Opinion in HIV and AIDS | Year: 2010

Purpose of review: To consider how nonhuman primate (NHP) model systems can best contribute to HIV vaccine development. Recent Findings: We review the traditional roles of NHP model systems in vaccine development and compare this with how NHP models have been used in HIV vaccine research and development. Comparisons of the immune responses elicited by cellular immune response-inducing vaccines in macaques and humans illustrate the value of primate studies for the relative ranking of HIV vaccine concepts for their likely immunogenicity in humans. The unusual structures (e.g. long complementarity-determining regions) of known broadly neutralizing HIV antibodies (bNAbs) suggest that it is critical to test candidate env immunogens in NHPs, whose germline antibody repertoires resemble those of humans. Recent clinical efficacy trial results question the utility of existing NHP challenge models in predicting HIV vaccine efficacy in humans, and highlight the need to further develop models in which acquisition of infection can be reliably evaluated. When evaluated in models using low virus dose challenges that better approximate human sexual exposure to HIV - some vaccine and passive NAb interventions appear to protect against acquisition of infection. Summary: NHP models have important roles in the preclinical evaluation, optimization, and ranking of novel HIV immunogens. The apparent vaccine efficacy observed using low virus dose challenge models provides an opportunity to investigate the correlates of protection. © 2010 Wolters Kluwer Health | Lippincott Williams & Wilkins.

Since publication of a 1997 review of the immunogenicity and safety data for pneumococcal polysaccharide vaccines (PPSVs), dozens of additional studies have been published, involving larger cohorts, longer observation periods, and more specific assays. Additionally, a 13-valent pneumococcal conjugate vaccine (PCV) has been licensed for adults. This paper reviews adult studies assessing antibody persistence for ≥3 years after pneumococcal vaccination, and adult studies of immunogenicity and safety after revaccination. This review emphasizes the currently registered PPSV23 formulations containing 25-μg polysaccharide per serotype, for which far more long-term data are available. Broadly, IgG and functional antibody levels after PPSV23 in adults persist above concentrations in unvaccinated adults for at least 5-10 years in most studies. The few exceptions involve populations of non-ambulatory adults or those with confounding host-factor issues. Revaccination with PPSV23 5-10 years after a previous dose consistently and substantially increases both IgG and functional antibody levels. There is an inverse association between circulating antibody level just before primary or revaccination and subsequent antibody increase. Although injection-site reactions (e.g., pain, swelling, redness) were reported more commonly after PPSV23 revaccination than after primary vaccination in most studies, these reactions typically resolved within 5 days. We interpret the contemporary literature as supporting pneumococcal revaccination as a means to sustain anti-pneumococcal antibodies at levels greater than among unvaccinated adults. PPSV23 is a broad-spectrum public-health tool to help prevent serious pneumococcal diseases across the adult lifespan. © 2012 Elsevier Ltd.

Wu N.,Merck And Co. | Bradley A.C.,North Carolina A&T State University | Welch C.J.,Merck And Co. | Zhang L.,Merck And Co.
Journal of Separation Science | Year: 2012

Effects of extra-column volume on apparent separation parameters were studied in ultra-high pressure liquid chromatography with columns and inlet connection tubings of various internal diameters (id) using 50-mm long columns packed with 1.8-μm particles under isocratic conditions. The results showed that apparent retention factors were on average 5, 11, 18, and 41% lower than those corrected with extra-column volumes for 4.6-, 3.0-, 2.1-, and 1.0-mm id columns, respectively, when the extra-column volume (11.3 μL) was kept constant. Also, apparent pressures were 31, 16, 12, and 10% higher than those corrected with pressures from extra-column volumes for 4.6-, 3.0-, 2.1-, and 1.0-mm id columns at the respective optimum flow rate for a typical ultra-high pressure liquid chromatography system. The loss in apparent efficiency increased dramatically from 4.6- to 3.0- to 2.1- to 1.0-mm id columns, less significantly as retention factors increased. The column efficiency was significantly improved as the inlet tubing id was decreased for a given column. The results suggest that maximum ratio of extra-column volume to column void volume should be approximately 1:10 for column porosity more than 0.6 and a retention factor more than 5, where 80% or higher of theoretically predicted efficiency could be achieved. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Leucht S.,TU Munich | Fennema H.,Biostatistics and Research Decision science MSD | Engel R.,Ludwig Maximilians University of Munich | Kaspers-Janssen M.,Biostatistics and Research Decision science MSD | And 3 more authors.
Journal of Affective Disorders | Year: 2013

Little is known about the clinical relevance of the Hamilton Rating Scale for Depression (HAMD-17) total scores. It is unclear how total scores translate into clinical severity, or what commonly used measures for response (reduction from baseline of ≥50% in the total score) and remission (total HAMD-17 score ≤7) mean from a clinical perspective. We therefore compared: (a) the percentage and absolute change in the HAMD-17 total scores with Clinical Global Impression-Improvement (CGI-I); (b) the absolute and percentage change in the HAMD-17 total scores with Clinical Global Impression-Severity (CGI-S) absolute change; and (c) the percentage and absolute change in the HAMD-17 total scores with CGI-I in the subgroups of patients with≤median and>median HAMD-17 total scores at baseline. The method used was equipercentile linking of HAMD-17 and CGI ratings from 43 drug trials in patients with Major Depressive Disorder (MDD) (n=7131). Our results confirm the validity of the commonly used measures for remission and response in MDD trials: a CGI-I score of 2 ('much improved') corresponded to a reduction from baseline of >50% and <60%, and a CGI-I score of 1 ('very much improved') to a reduction of >75% and <85%. The CGI-S score of 1 ('normal,; not at all ill') corresponded to the HAMD-17 total score of <5 and the CGI-S score of 2 ('borderline mentally ill') to the score between 6 and 8. An effect of baseline illness severity was observed. © 2012 Elsevier B.V.

Shentu Y.,Rutgers University | Shentu Y.,Merck And Co. | Xie M.,Rutgers University
Statistics in Medicine | Year: 2010

The purpose of this note is to raise awareness of the complexity of the practice involving dichotomization. It is well known that the regular regression models are effective tools for analyzing Gaussian-type response variables, and researchers are often told that it is a 'bad idea' to practice dichotomization if continuous measurements are available. We demonstrate through special cases, however, that there is another side of the story if the response variable is contaminated. Although dichotomization causes loss of information, it can also reduce input of contamination. If the reduction of contamination input outweighs the loss of information, analysis based on dichotomization can sometimes provide better results. We derive formulas of bias and variance for binary regression estimators under a contamination model of unknown additive errors, and compare them with both the least squares and robust M-estimators from the corresponding linear regression analysis using continuous responses. As a case study, we study extensively the case in which the observed response is contaminated by an error with a mean and a variance proportional to the mean and the variance of the uncontaminated true response. Conditions under which dichotomization is preferred are obtained. A simulation study based on a real data setting is provided, which supports the theoretical developments. Copyright © 2010 John Wiley & Sons, Ltd.

Verkaar F.,Merck And Co. | Verkaar F.,Maastricht University | Zaman G.J.R.,Merck And Co.
FEBS Letters | Year: 2010

Wnts control mammalian developmental morphogenesis and are critical for adult stem cell maintenance. Wnts initiate several intracellular signaling cascades, such as Wnt/β-catenin-, Wnt/Ca2+- and Wnt/ROR2-signaling. Signaling preference of Wnts for these various pathways is thought to depend on the repertoire of receptors present on recipient cells. Here, we propose a further refinement of this receptor model and hypothesize that Wnt signaling specificity depends on co-receptor recruitment upon binding of Wnt to Frizzled receptor molecules. In this model, recruitment of LRP5/6 leads to activation of Wnt/β-catenin signaling, whereas signaling through other pathways is mediated by recruiting ROR2. © 2010 Federation of European Biochemical Societies.

Chen B.,Indiana University Bloomington | Sheridan R.P.,Merck And Co. | Hornak V.,Merck And Co. | Voigt J.H.,Merck And Co.
Journal of Chemical Information and Modeling | Year: 2012

Random forest is currently considered one of the best QSAR methods available in terms of accuracy of prediction. However, it is computationally intensive. Naïve Bayes is a simple, robust classification method. The Laplacian-modified Naïve Bayes implementation is the preferred QSAR method in the widely used commercial chemoinformatics platform Pipeline Pilot. We made a comparison of the ability of Pipeline Pilot Naïve Bayes (PLPNB) and random forest to make accurate predictions on 18 large, diverse in-house QSAR data sets. These include on-target and ADME-related activities. These data sets were set up as classification problems with either binary or multicategory activities. We used a time-split method of dividing training and test sets, as we feel this is a realistic way of simulating prospective prediction. PLPNB is computationally efficient. However, random forest predictions are at least as good and in many cases significantly better than those of PLPNB on our data sets. PLPNB performs better with ECFP4 and ECFP6 descriptors, which are native to Pipeline Pilot, and more poorly with other descriptors we tried. © 2012 American Chemical Society.

Kawai K.,Merck And Co. | Gebremeskel B.G.,Rutgers University | Acosta C.J.,Merck And Co.
BMJ Open | Year: 2014

Objective: The objective of this study was to characterise the incidence rates of herpes zoster (HZ), also known as shingles, and risk of complications across the world. Design: We systematically reviewed studies examining the incidence rates of HZ, temporal trends of HZ, the risk of complications including postherpetic neuralgia (PHN) and HZ-associated hospitalisation and mortality rates in the general population. The literature search was conducted using PubMed, EMBASE and the WHO library up to December 2013. Results: We included 130 studies conducted in 26 countries. The incidence rate of HZ ranged between 3 and 5/1000 person-years in North America, Europe and Asia-Pacific, based on studies using prospective surveillance, electronic medical record data or administrative data with medical record review. A temporal increase in the incidence of HZ was reported in the past several decades across seven countries, often occurring before the introduction of varicella vaccination programmes. The risk of developing PHN varied from 5% to more than 30%, depending on the type of study design, age distribution of study populations and definition. More than 30% of patients with PHN experienced persistent pain for more than 1 year. The risk of recurrence of HZ ranged from 1% to 6%, with long-term follow-up studies showing higher risk (5-6%). Hospitalisation rates ranged from 2 to 25/100 000 person-years, with higher rates among elderly populations. Conclusions: HZ is a significant global health burden that is expected to increase as the population ages. Future research with rigorous methods is important.

Modi A.,Merck And Co. | Siris E.S.,Columbia University | Tang J.,Asclepius Analytics Ltd. | Sen S.,Merck And Co.
Current Medical Research and Opinion | Year: 2015

Objective: The objective was to evaluate compliance with osteoporosis (OP) treatments and determine the fracture and healthcare burden associated with noncompliance. Methods: This retrospective analysis of a US claims database identified women initiating an OP medication from 1 January 2002 to 30 June 2009. Patients were ≥55 years and had ≥1 pharmacy claim for a bisphosphonate or non-bisphosphonate (raloxifene, calcitonin, teriparatide); the index date was the first pharmacy claim. There were three study periods: baseline (12 months pre-index); compliance period (0-12 months post-index); and follow-up period (12-24 months post-index). Medication possession ratio (MPR) was calculated during the compliance period to differentiate two cohorts: compliant (MPR≥80%) and noncompliant (MPR<80%). Outcomes during follow-up were modeled by logistic regression (presence of fracture), Poisson regression (healthcare utilization incidence rate) and gamma regression (healthcare costs), all adjusted for patient demographic and clinical characteristics. Results: Overall, 685,505 women initiating OP therapy were identified and 57,913 (8.4%) met the inclusion criteria: only 23,430 (40.5%) were compliant and 34,483 (59.5%) were noncompliant. Mean age was 64 years. Noncompliance was associated with a 20% higher risk of any fracture (odds ratio: 1.20, 95% CI=1.07-1.35), a higher incidence rate ratio (IRR) for inpatient utilization (IRR: 1.26, 95% CI=1.19-1.34) and a lower rate of outpatient utilization (IRR: 0.97, 95% CI=0.95-0.98). Noncompliant patients had 13% higher medical costs (cost ratio: 1.13, 95% CI=1.06-1.21) than compliant patients. Limitations: Inclusion in this study required 36 months of continuous healthcare coverage. Thus, the results are primarily applicable to a stable, managed care population and may not be generalizable to other populations. Conclusion: Noncompliance with OP therapy was associated with a higher risk of fracture, higher all-cause medical costs and a higher frequency of inpatient service utilization. Additional research is needed to identify barriers to compliance with OP therapy. © 2015 All rights reserved: reproduction in whole or part not permitted.

Brixen K.,University of Southern Denmark | Chapurlat R.,University of Lyon | Cheung A.M.,University of Toronto | Keaveny T.M.,University of California at Berkeley | And 9 more authors.
Journal of Clinical Endocrinology and Metabolism | Year: 2013

Context: Odanacatib, a cathepsin K inhibitor, increases spine and hip areal bone mineral density (BMD)in postmenopausal women with low BMD and cortical thickness in ovariectomized monkeys. Objective: The objective of the study was to examine the impact of odanacatib on the trabecular and cortical bone compartments and estimated strength at the hip and spine. Design: This was a randomized, double-blind, 2-year trial. Setting: The study was conducted at a private or institutional practice. Participants: Participants included 214 postmenopausal women with low areal BMD. Intervention: The intervention included odanacatib 50 mg or placebo weekly. Main Outcome Measures: Changes in areal BMD by dual-energy x-ray absorptiometry (primary end point, 1 year areal BMD change at lumbar spine), bone turnover markers, volumetric BMD by quantitative computed tomography (QCT), and bone strength estimated by finite element analysis were measured. Results: Year 1 lumbar spine areal BMD percent change from baseline was 3.5% greater with odanacatib than placebo (P<.001). Bone-resorption marker C-telopeptide of type 1 collagen was significantly lower with odanacatib vs placebo at 6 months and 2 years (P<.001). Bone-formation marker procollagen I N-terminal peptide initially decreased with odanacatib but by 2 years did not differ from placebo. After 6 months, odanacatib-treated women had greater increases in trabecular volumetric BMD and estimated compressive strength at the spine and integral and trabecular volumetric BMD and estimated strength at the hip (P<.001). At the cortical envelope of the femoral neck, bone mineral content, thickness, volume, and cross-sectional area also increased from baseline with odanacatib vs placebo (P<.001 at 24 months). Adverse experiences were similar between groups. Conclusions: Over 2 years, odanacatib decreased bone resorption, maintained bone formation, increased areal and volumetric BMD, and increased estimated bone strength at both the hip and spine. Copyright © 2013 by The Endocrine Society.

Gunaydin H.,Merck And Co. | Bartberger M.D.,Amgen
ACS Medicinal Chemistry Letters | Year: 2016

This viewpoint describes the results obtained from matched molecular pair analyses and quantum mechanics calculations that show unsaturated rings found in drug-like molecules may be replaced with their saturated counterparts without losing potency even if they are engaged in stacking interactions with the side chains of aromatic residues. © 2016 American Chemical Society.

Peterson E.A.,Amgen | Dillon B.,Astrazeneca | Raheem I.,Merck And Co. | Richardson P.,Pfizer | And 3 more authors.
Green Chemistry | Year: 2014

Chromatography is routinely used in drug discovery as a means to isolate intermediates and final compounds. From a sustainability perspective, it is one of the largest contributors of solvent waste in the drug discovery process. The medicinal chemistry subgroup within the American Chemical Society's Green Chemistry Institute Pharmaceutical Roundtable (ACS GCI PR) offers a perspective aimed at providing chemists with practical tools and easily implemented techniques to improve the sustainability of drug discovery through reduction of the waste generated during chromatography. This perspective also offers alternatives to traditional, silica gel-based chromatography as well as information on how to avoid chromatography completely through use of crystallization and reaction telescoping. © the Partner Organisations 2014.

Martin G.E.,Merck And Co. | Sunseri D.,External Manufacturing Drug Delivery Systems
Journal of Pharmaceutical and Biomedical Analysis | Year: 2011

Two-dimensional NMR methods are the cornerstone of modern structure elucidation methods. When the ensemble of 1D and 2D NMR experiments normally employed for structure assignment fails, investigators typically resort to successively more complex 2D NMR experiments for structure determination and/or spectral assignment. Unsymmetrical Indirect Covariance (UIC) NMR data processing methods provide a convenient and highly efficient means of accessing the connectivity information embodied in more complex experiments such as HSQC-TOCSY spectra. Using Unsymmetrical Indirect Covariance (UIC) or General Indirect Covariance (GIC) processing to mathematically combine multiplicity-edited GHSQC and 1,1-ADEQUATE 2D NMR spectra affords an HSQC-ADEQUATE spectrum that offers a new method for establishing the carbon skeleton of a molecule. The application of this technique is demonstrated for a novel cyclin-dependant kinase inhibitor, Dinaciclib™ (SCH 727965). © 2011 Elsevier B.V.

Chen J.,Abbott Laboratories | Heyse J.F.,Merck And Co. | Heaton P.,Novartis | Kuter B.J.,Merck Vaccines
American Journal of Epidemiology | Year: 2010

The relation between the risk of intussusception and age at the time of receipt of the first dose of rhesus-human reassortant rotavirus tetravalent vaccine (RRV-TV) has been studied extensively on the basis of Centers for Disease Control and Prevention (CDC) matched case-control study data, using various statistical methods, including conditional logistic regression and quadratic smoothing splines. However, different conclusions have been reported in published analyses regarding the dependence of the risk of intussusception on age at first dose. The authors reanalyzed the CDC matched case-control data set using unrestricted and restricted quadratic smoothing spline methods for various exposure windows (i.e., intervals postvaccination). These analyses indicated that the use of different models may lead to different conclusions. The restricted quadratic smoothing spline with appropriately chosen knot locations showed a statistically significant increased risk of intussusception associated with RRV-TV for the exposure window 3-14 days after the first dose at an age as young as 49 days, the youngest age in the data set at which vaccine was administered; this implies an increased risk of intussusception associated with RRV-TV at all ages studied. © The Author 2010.