Genevini P.,University of Milan |
Papiani G.,University of Milan |
Papiani G.,Oligomerix, Inc |
Ruggiano A.,University of Milan |
And 5 more authors.
PLoS ONE | Year: 2014
VAPB is a ubiquitously expressed, ER-resident adaptor protein involved in interorganellar lipid exchange, membrane contact site formation, and membrane trafficking. Its mutant form, P56S-VAPB, which has been linked to a dominantly inherited form of Amyotrophic Lateral Sclerosis (ALS8), generates intracellular inclusions consisting in restructured ER domains whose role in ALS pathogenesis has not been elucidated. P56S-VAPB is less stable than the wild-type protein and, at variance with most pathological aggregates, its inclusions are cleared by the proteasome. Based on studies with cultured cells overexpressing the mutant protein, it has been suggested that VAPB inclusions may exert a pathogenic effect either by sequestering the wild-type protein and other interactors (loss-of-function by a dominant negative effect) or by a more general proteotoxic action (gain-of-function). To investigate P56S-VAPB degradation and the effect of the inclusions on proteostasis and on ER-to-plasma membrane protein transport in a more physiological setting, we used stable HeLa and NSC34 Tet-Off cell lines inducibly expressing moderate levels of P56S-VAPB. Under basal conditions, P56S-VAPB degradation was mediated exclusively by the proteasome in both cell lines, however, it could be targeted also by starvation-stimulated autophagy. To assess possible proteasome impairment, the HeLa cell line was transiently transfected with the ERAD (ER Associated Degradation) substrate CD3δ, while autophagic flow was investigated in cells either starved or treated with an autophagy-stimulating drug. Secretory pathway functionality was evaluated by analyzing the transport of transfected Vesicular Stomatitis Virus Glycoprotein (VSVG). P56S-VAPB expression had no effect either on the degradation of CD3δ or on the levels of autophagic markers, or on the rate of transport of VSVG to the cell surface. We conclude that P56S-VAPB inclusions expressed at moderate levels do not interfere with protein degradation pathways or protein transport, suggesting that the dominant inheritance of the mutant gene may be due mainly to haploinsufficiency. © 2014 Genevini et al. Source
Gao Z.,Sanofi S.A. |
Hurst W.J.,Sanofi S.A. |
Czechtizky W.,Sanofi S.A. |
Francon D.,Sanofi S.A. |
And 8 more authors.
Bioorganic and Medicinal Chemistry Letters | Year: 2013
Previous studies have shown that compound 1 displayed high affinity towards histamine H3 receptor (H3R), (human (h-H3R), Ki = 8.6 nM, rhesus monkey (rh-H3R), Ki = 1.2 nM, and rat (r-H3R), Ki = 16.5 nM), but exhibited high affinity for hERG channel. Herein, we report the discovery of a novel, potent, and highly selective H3R antagonist/inverse agonist 5a(SS) (SAR110068) with acceptable hERG channel selectivity and desirable pharmacological and pharmacokinetic properties through lead optimization sequence. The significant awakening effects of 5a(SS) on sleep-wake cycles studied by using EEG recording in rats during their light phase support its potential therapeutic utility in human sleep-wake disorders. © 2013 Elsevier Ltd. All rights reserved. Source
Oligomerix, Inc | Date: 2013-03-26
Chemical products and compositions intended for medical research, namely, reagents, chemical entities and compounds; chemical preparations for scientific purposes, chemical reagents or products for laboratory analyses for monitoring, analyzing and identifying diagnostic targets and therapeutic products, and for conducting research in connection with pharmaceutical sciences; chemical preparations intended for scientific purposes, namely, for leading research in connection with pharmaceutical sciences. Pharmaceutical products for the treatment of diseases, namely, Alzheimers disease and other neurodegenerative diseases; reagents for diagnostic purposes and diagnostic media intended for medical purposes, namely, reagents and media intended for tests, namely, tests based on blood biomarkers for identifying patients affected by diseases or those at risk of developing a disease and for identifying patients responding to specific personalized treatments; biological acidic products and preparations for pharmaceutical purposes, namely, pharmaceutical preparations for diagnosing diseases, namely, Alzheimers disease and other neurodegenerative diseases; chemico-pharmaceutical preparations for diagnosing, preventing and treating diseases, namely, Alzheimers disease and other neurodegenerative diseases; biopharmaceutical products, namely, reagents, chemical entities and compounds for diagnosing, preventing and treating diseases, namely, Alzheimers disease and other neurodegenerative diseases; chemical preparations for medical and pharmaceutical purposes, namely, chemicals for diagnosing Alzheimers disease and other neurodegenerative diseases, especially chemical preparations for diagnosing diseases in the nature of neurodegenerative diseases. Research and development of pharmaceutical molecules, compounds, inhibitors, antibodies and biological markers for diagnostic purposes and medicines for treating diseases; research and development of new products and preparations for the treatment of Alzheimers disease and other neurodegenerative diseases; research and development in the field of pharmaceutical drugs; biological research; biotechnological research; technical research in the field of biotechnology; consultancy in the field of biotechnology.
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 323.29K | Year: 2009
DESCRIPTION (provided by applicant): There is a critical unmet need for novel biomarkers for differential diagnosis of Alzheimer's disease (AD) at early onset and to stage disease progression. Presently, definitive diagnosis of AD requires tissue biopsy or autopsy of the brain, which can occur only after the patient dies. Currently, there are no validated biomarkers identified in patient samples such as cerebrospinal fluid (CSF), blood, and urine that can be used to definitely monitor the progression or regression of AD. Most approaches focus on quantitative changes of individual proteins such as tau and A2 in patient samples as biomarkers. These studies have led to a consensus that an increase in total and p-tau and a concomitant decrease in A21-42 in CSF may be indicative of neurodegeneration. However, these changes are not specific indicators of AD and may also occur in some other forms of dementia. Therefore, these biomarkers cannot be used for differential diagnosis of AD at early onset and to stage disease progression. The development of novel biomarkers based on the paradigm of protein-protein interactions, rather than on specific protein levels, may create opportunity for earlier and more specific diagnosis and disease monitoring for better patient management. Using the approach of measuring pathological interactions between proteins may provide urgently needed novel AD biomarkers. These biomarkers can also serve as drug targets for the identification of new medication therapies to treat AD and to monitor different therapeutic effects when used to treat AD. OLIGOMERIX is evaluating novel AD biomarkers based on soluble complexes containing tau oligomers and complexes containing both tau and A21-42. Preliminary feasibility studies with a limited number of CSF samples have indicated that tau oligomers and tau- A21-42 complexes are enhanced in CSF from AD compared to non-AD controls. ELISAs of different formats were developed to measure these parameters in CSF. The objective of this project is to expand the preliminary studies using a statistically significant number of clinical lumbar CSF samples that include severe, mild and age-matched non-AD controls to assess the potential of tau oligomers tau A2, and tau-apoE complexes as specific biomarkers for early detection and progression of AD. This work will be performed in collaboration with Drs. Richard Mayeux and Lawrence Honig at the Taub Institute for Research on Alzheimer's disease and the Aging Brain at Columbia University Medical Center. The specific aims of the proposed program are as follows: Develop biomarker assays for AD measuring tau oligomers and complexes of tau with A2 or apoE in CSF Conduct feasibility study of developed biomarker assays using clinical CSF samples Towards this end Dr. Honig will provide about 250 CSF samples for a statistically significant study of the biomarkers. OLIGOMERIX will perform ELISAs on de-identified blinded samples, and Dr. Honig will analyze the data. The biomarkers may be out-licensed to the pharmaceutical industry for clinical evaluation of compounds targeting tau oligomers, tau-A2, or tau-apoEcomplexes. Key Words Alzheimer's disease, biomarker, tau, A2, apoE, oligomer. PUBLIC HEALTH RELEVANCE: The proposed program is focused on the identification and development of novel biomarkers for Alzheimer's disease (AD), based on the paradigm of protein-protein interactions. Presently, AD biomarker studies are primarily focused on the quantitative changes in the levels of protein involved in AD etiology. In addition to AD, these changes may also occur in some other forms of dementia, therefore, are not specific indicators of AD. Here we propose to evaluate novel AD biomarkers based on soluble complexes containing key protein players in AD: tau, A21-42 and apoE. Our goal is to 1) develop biomarker assays to measure the interactions of tau protein with itself (tau oligomers), A2 or apoE in human CSF samples, 2) evaluate the feasibility of using tau oligomers, or tau complexes with A2 or apoE as biomarkers for AD using a significantly large number of CSF samples.
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase II | Award Amount: 1.65M | Year: 2010
DESCRIPTION (provided by applicant): High Throughput Tau Oligomer Assay for Drug Screening for Alzheimer's Disease Project Summary: There is a large and rapidly growing unmet need for disease modifying drugs for Alzheimer's disease. Currently there are 18 million cases of AD worldwide; by 2025 this number is expected to increase to 34 million. Presently, only 5 mildly effective AD symptom-treating drugs exist, but none that treat the underlying neurodegenerative processes. Tau is becoming a more prominent target for the development of disease- modifying drugs (DMDs), as its role in neurodegeneration is becoming better understood. Mutations in the gene for tau protein MAPT are causative of dementia and tau pathology correlates well with AD progression. At the same time, late stage clinical failures for therapeutics based on the amyloid hypothesis have raised questions on solely targeting A2. Strong evidence has emerged implicating tau oligomers as playing a direct role in disease pathogenesis for AD and over 20 other neurodegenerative diseases (Brunden et al. 2008; Davidowitz et al. 2008). To discover drugs targeting tau oligomerization methods were developed to select compounds inhibiting tau self-interaction, and an assay using AlphaScreen detection technology was selected for further development for high throughput screening (Chatterjee et al. 2008). In addition, the phage display- atomic force microscopy method developed by Dr. Sierks (ASU) was used to isolate antibody fragments (scFvs) specifically binding to tau oligomers that will be adopted in the cell based screening assays. The specific aims of the proposed program are as follows: Convert the tau oligomer assay to HTS format Transfer Assay to the Michigan High Throughput Screening Center (MHTSC) for automation and screening of a highly optimized compound library (100,000 compounds) and carry out medicinal chemistry analysis to model the pharmacophore and select additional chemotypes for screening Select compounds using tau oligomer specific antibody fragments in cell based assays The anticipated outcome of the proposed Phase II program is the selection of at least three or more lead candidate compounds targeting tau oligomers that will be developed during the Phase III program and evaluated in animal models of AD and tauopathies. To attain this result, the high throughput assay will be optimized and transferred to the Michigan High Throughput Screening Center where their Select Set library of approximately 100,000 compounds will be screened under the direction Dr. Robert Kilkuskie Hits will be validated and a structural pharmacaophore model will be developed. Toxic compounds will be eliminated using a neurocytotoxicity assay. Three or more scFvs will be selected with high sensitivity and specificity that will be used to identify compounds inhibiting tau self-association using in vitro cell based assays. Antibody fragments, in addition to enabling the primary goal, are supportive of the company's other programs including its tau biomarker development program and its tau immunotherapeutic program. However, the primary goal of the program is to advance the tau oligomer drug discovery platform to identify active inhibitors as lead candidates for IND enabling studies. Key Words Alzheimer's disease, tauopathy, neurodegenerative disease, drug discovery, tau oligomer, phage display, antibody fragment, scFv, high throughput screening PUBLIC HEALTH RELEVANCE: The proposed program focuses on developing a high throughput screening assay targeting tau oligomers for drug discovery for Alzheimer's disease (AD). This project was inspired by observations that accumulation of tau oligomers has been shown to correlate well with neuronal loss and memory impairment in AD and in tauopathy mouse models. OLIGOMERIX has developed in vitro assays for screening compounds that inhibit the formation of cytotoxic tau oligomers. This program aims to 1.) .Convert the tau oligomer assay to HTS format; 2.) Transfer Assay to the Michigan High Throughput Screening Center (MHTSC) for automation and screening of a highly optimized compound library (100,000 compounds) and carry out medicinal chemistry analysis to model the pharmacophore; 3.) Select compounds using tau oligomer specific antibody fragments and cell based screening to identify lead candidates for future animal studies and pre-clinical development.