Lin Q.,Vanderbilt University |
Lin Q.,Sundia MediTech |
Liu Y.,Vanderbilt University |
Moore D.J.,Vanderbilt University |
And 4 more authors.
Journal of Immunology | Year: 2012
The expression of proinflammatory cytokines and chemokines in response to TCR agonists is regulated by the caspase-recruitment domain membrane-associated guanylate kinase 1 (CARMA1) signalosome through the coordinated assembly of complexes containing the BCL10 adaptor protein. We describe a novel mechanism to negatively regulate the CARMA1 signalosome by the "death" adaptor protein caspase and receptor interacting protein adaptor with death domain (CRADD)/ receptor interacting protein-associated ICH-1/CED-3 homologous protein with a death domain. We show that CRADD interacts with BCL10 through its caspase recruitment domain and suppresses interactions between BCL10 and CARMA1. TCR agonist-induced interaction between CRADD and BCL10 coincides with reduction of its complex formation with CARMA1 in wild-type, as compared with Cradd-deficient, primary cells. Finally, Cradd-deficient spleen cells, CD4 +T cells, and mice respond to T cell agonists with strikingly higher production of proinflammatory mediators, including IFN-γ, IL-2, TNF-α, and IL-17. These results define a novel role for CRADD as a negative regulator of the CARMA1 signalosome and suppressor of Th1-and Th17-mediated inflammatory responses. Copyright © 2012 by The American Association of Immunologists, Inc.
Luo D.,Tongji University |
Shi Y.,Sundia MediTech |
Wang J.,Sundia MediTech |
Lin Q.,Sundia MediTech |
And 5 more authors.
Neuroscience Letters | Year: 2016
Brain-derived neurotrophic factor (BDNF) is a notably important neurotrophin which regulates neuronal survival and differentiation in the nervous system. However, its clinical usage is particularly limited. 7,8-dihydroxyflavone (7,8-DHF), which acts as a selective agonist of BDNF receptor TrkB, is reported to possess neuroprotective effects both in vitro and in vivo. Here we explored the potent neuroprotective effects of 7,8-DHF in 6-OHDA induced rat and MPTP induced mouse model of Parkinsonism. The results demonstrated that treatment with 7,8-DHF in drinking water for four weeks (two weeks before 6-OHDA + two weeks after 6-OHDA lesion) significantly improved dopamine-mediated behaviors in 6-OHDA rat model, and prevented the loss of dopaminergic neurons in the substantia nigra (SN). Phospho-Y816-TrkB immunostaining showed that TrkB phosphorylation was significantly elevated in the SN in 7,8-DHF pretreated group, indicating 7,8-DHF activated TrkB and likely contributed to its neuroprotective effects. 7,8-DHF also protected acute MPTP neurotoxicity in mice but did not affect the climbing behavior in pole test. Thus our study indicates the neuroprotective properties of 7,8-DHF through the activation of TrkB, which provides a novel therapeutic treatment for Parkinson's disease. © 2016 Elsevier Ireland Ltd.
Meng L.,Fudan University |
Shu M.,Shanghai JiaoTong University |
Chen Y.,Fudan University |
Yang D.,Sundia MediTech |
And 5 more authors.
Cancer Biology and Therapy | Year: 2014
The anaplastic lymphoma kinase (ALK) and the c-Met receptor tyrosine kinase play essential roles in the pathogenesis in multiple human cancers and present emerging targets for cancer treatment. Here, we describe CM-118, a novel lead compound displaying low nanomolar biochemical potency against both ALK and c-Met with selectivity over >90 human kinases. CM-118 potently abrogated hepatocyte growth factor (HGF)-induced c-Met phosphorylation and cell migration, phosphorylation of ALK, EML4-ALK, and ALK resistance mutants in transfected cells. CM-118 inhibited proliferation and/or induced apoptosis in multiple c-Met- and ALK-addicted cancer lines with dose response profile correlating target blockade. We show that the CM-118-induced apoptosis in c-Met-amplified H1993 NSCLC cells involved a rapid suppression of c-Met activity and c-Met-to-EGFR cross-talk, and was profoundly potentiated by EGFR inhibitors as shown by the increased levels of apoptotic proteins cleaved-PARP and Bim as well as reduction of the survival protein Mcl-1. Bim-knockdown or Mcl-1 overexpression each significantly attenuated apoptosis. We also revealed a key role by mTOR in mediating CM-118 action against the EML4-ALK-dependent NSCLC cells. Abrogation of EML4-ALK in H2228 cells profoundly reduced signaling capacity of the rapamycin-sensitive mTOR pathway leading to G1 cell cycle arrest and mitochondrial hyperpolarization, a metabolic perturbation linked to mTOR inhibition. Depletion of mTOR or mTORC1 inhibited H2228 cell growth, and mTOR inhibitors potentiated CM-118's antitumor activity in vitro and in vivo. Oral administration of CM-118 at a wide range of well tolerated dosages diminished c-Met- and ALK phosphorylation in vivo, and caused tumor regression or growth inhibition in multiple c-Met- and ALK-dependent tumor xenografts in mice. CM-118 exhibits favorable pharmacokinetic and drug metabolism properties hence presents a candidate for clinical evaluation. © 2014 Landes Bioscience.
Chen A.,Central China Normal University |
Bao Y.,Sundia MediTech |
Ge X.,Central China Normal University |
Shin Y.,Pacific Northwest National Laboratory |
And 2 more authors.
RSC Advances | Year: 2012
Phosphorylated p53 at serine 15 (phospho-p5315) is a potential biomarker of gamma-radiation exposure. In this paper, we described a new magnetic particle (MP)-based electrochemical immunoassay of human phospho-p5315 using carbon nanospheres (NS) and protein cage nanoparticles (PCN) for signal amplification. Greatly enhanced sensitivity was achieved for three reasons: 1) PCN and the p5315 signal antibody (p5315 Ab2) are linked to the carbon NS (PCN-p53 15 Ab2-NS) as a label; 2) PCN increases the amount of metal ions in the cavity of each apoferritin; 3) MPs capture a large amount of primary antibodies. Protein cage templated metallic phosphates, instead of enzymes, as multi-labels have the advantage of eliminating the addition of mediator or immunoreagents and, thus, makes the immunoassay system simpler. Subsequent stripping voltametric analysis, detected olead ions on a disposable screen-printed electrode. The response current was proportional to the phospho-p5315 concentration in the range of 0.02 to 20 ng mL -1 with a detection limit of 0.01 ng mL-1, which was 30-fold lower than that of the ELISA measurement of phospho-p5315. This method shows an acceptable stability and reproducibility and the assay results for phospho-p5315-spiked human serum presented good recovery rates. © 2012 The Royal Society of Chemistry.
Bao Y.,Sundia MediTech |
Wang Q.,Suzhou Kangrun Pharmaceutical Testing Service Inc. |
Tang P.,Suzhou Kangrun Pharmaceutical Testing Service Inc.
Journal of Mass Spectrometry | Year: 2013
A novel, rapid and sensitive liquid chromatography/quadrupole linear ion trap mass spectrometry [LC-ESI-(QqLIT)MS/MS] method was developed and validated for the quantification of protopanaxadiol (PPD) in rat plasma. Oleanolic acid (OA) was used as internal standard (IS). A simple protein precipitation based on acetonitrile (ACN) was employed. Chromatographic separation was performed on a Sepax GP-C18 column (50 × 2.1 mm, 5 μM) with a mobile phase consisting of ACN-water and 1.5 μM formic acid and 25 mM lithium acetate (90: 10, v/v) at a flow rate of 0.4 ml/min for 3.0 min. Multiple-reaction-monitoring mode was performed using lithium adduct ion as precursor ion of m/z 467.5/449.4 and 455.6/407.4 for the drug and IS, respectively. Calibration curve was recovered over a concentration range of 0.5-100 ng/ml with a correlation coefficient >0.99. The limit of detection was 0.2 ng/ml in rat plasma for PPD. The results of the intraday and interday precision and accuracy studies were well within the acceptable limits. The validated method was successfully applied to investigate the pharmacokinetic study of PPD after intravenous and gavage administration to rat. Copyright © 2013 John Wiley & Sons, Ltd.
Karasawa H.,University of California at Los Angeles |
Pietra C.,Helsinn SA Lugano |
Giuliano C.,Helsinn SA Lugano |
Garcia-Rubio S.,Helsinn Therapeutics U.S. Inc. |
And 4 more authors.
Neurogastroenterology and Motility | Year: 2014
Background: Constipation and L-dopa-induced gastric dysmotility are common gastrointestinal (GI) symptoms in Parkinson's disease (PD). We investigated the novel ghrelin agonist, HM01 influence on GI motor dysfunctions in 6-hydroxydopamine (6-OHDA) rats. Methods: HM01 pharmacological profiles were determined in vitro and in vivo in rats. We assessed changes in fecal output and water content, and gastric emptying (GE) in 6-OHDA rats treated with orogastric (og) HM01 and L-dopa/carbidopa (LD/CD, 20/2 mg/kg). Fos immunoreactivity (ir) cells in specific brain and lumbosacral spinal cord were quantified. Key Results: HM01 displayed a high binding affinity to ghrelin receptor (Ki: 1.42 ± 0.36 nM), 4.3 ± 1.0 h half-life and high brain/plasma ratio. 6-OHDA rats had reduced daily fecal output (22%) and water intake (23%) compared to controls. HM01 (3 and 10 mg/kg) similarly reversed the decreased 4-h fecal weight and water content in 6-OHDA rats. Basal GE was not modified in 6-OHDA rats, however, LD/CD (once or daily for 8 days) delayed GE in 6-OHDA and control rats that was prevented by HM01 (3 mg/kg acute or daily before LD/CD). HM01 increased Fos-ir cell number in the area postrema, arcuate nucleus, nucleus tractus solitarius, and lumbosacral intermediolateral column of 6-OHDA rats where 6-OHDA had a lowering effect compared to controls. Conclusions & Inferences: 6-OHDA rats display constipation- and adipsia-like features of PD and L-dopa-inhibited GE. The new orally active ghrelin agonist, HM01 crosses the blood-brain barrier and alleviates these alterations suggesting a potential benefit for PD with GI disorders. © 2014 John Wiley & Sons Ltd.
Zhao C.,Texas A&M University |
Zhao C.,Sundia MediTech |
Mitchell T.A.,Texas A&M University |
Mitchell T.A.,Illinois State University |
And 3 more authors.
Journal of the American Chemical Society | Year: 2012
The ZnCl 2-mediated tandem Mukaiyama aldol lactonization (TMAL) reaction of aldehydes and thiopyridyl ketene acetals provides a versatile, highly diastereoselective approach to trans-1,2-disubstituted β-lactones. Mechanistic and theoretical studies described herein demonstrate that both the efficiency of this process and the high diastereoselectivity are highly dependent upon the type of ketene acetal employed but independent of ketene acetal geometry. Significantly, we propose a novel and distinct mechanistic pathway for the ZnCl 2-mediated TMAL process versus other Mukaiyama aldol reactions based on our experimental evidence to date and further supported by calculations (B3LYP/BSI). Contrary to the commonly invoked mechanistic extremes of [2+2] cycloaddition and aldol lactonization mechanisms, investigations of the TMAL process suggest a concerted but asynchronous transition state between aldehydes and thiopyridyl ketene acetals. These calculations support a boat-like transition state that differs from commonly invoked Mukaiyama "open" or Zimmerman-Traxler "chair-like" transition-state models. Furthermore, experimental studies support the beneficial effect of pre-coordination between ZnCl 2 and thiopyridyl ketene acetals prior to aldehyde addition for optimal reaction rates. Our previously proposed, silylated β-lactone intermediate that led to successful TMAL-based cascade sequences is also supported by the described calculations and ancillary experiments. These findings suggested that a similar TMAL process leading to β-lactones would be possible with an oxopyridyl ketene acetal, and this was confirmed experimentally, leading to a novel TMAL process that proceeds with efficiency comparable to that of the thiopyridyl system. © 2011 American Chemical Society.
Yang J.,Sundia MediTech
Proteomics Research Journal | Year: 2012
The effort to develop better description of protein three-dimensional folding structures has dominated biochemistry and drug discovery research for more than 70 years since Pauling first defined the helical configurations as secondary structure for protein in 1940. The challenge is how to acquire a complete description of protein folding shapes from N-terminal to C-terminal, including regular secondary structure as well as irregular tertiary structure. Here, a novel description method is introduced, which a set of 27 vectors is rigorously derived mathematically from an enclosed space. Each vector represents a three-dimensional folding shape of five successive Cα atoms, and the protein conformation can be completely described along protein backbone. These vectors are expressed by 27 alphabetic symbols, which are called as protein folding shape code (PFSC). Consequently, with PFSC, the folding conformation of any protein with given three-dimensional structure is able to be converted into a simple one-dimensional alphabetic string without gap. Furthermore, to take the advantage of one-dimensional description of folding shapes, the protein conformational structures are able to be compared with Needleman-Wunsch alignment algorithm. The global similarity of protein 3D structures is able to be assessed by a value of protein folding structure alignment score (PFSA-S) as a quantitative measurement, and the similarity and dissimilarity of local structures is able to be examined by alignment table. The results show that this approach has the capability not only to distinguish protein conformers with relatively high similarity, but also to compare proteins with diverse degrees in structural homology. Therefore, this approach provides a consistent procedure, and it produces a unique score for assessment of similarity in protein structure comparison. The significant is that the complete description of protein folding shapes provides a simple and effective means to screen protein database, compare protein structures, search protein fragment and probe drug binding site, study protein mutation and protein misfolding and so on. © 2012 Nova Science Publishers, Inc.
Zeng Y.,Sundia MediTech |
Zeng Y.,Shanghai University of Traditional Chinese Medicine |
Shao D.,Shanghai University of Traditional Chinese Medicine |
Fang Y.,East China Normal University
Analytical Letters | Year: 2011
An on-line two-dimension microflow liquid chromatography was developed for better separation and analysis of the highly complex ingredients of medicinal preparation of traditional Chinese medicine Coptis Chinensis Franch. A two-valve switching system was utilized for two-dimension chromatography with strong cation exchange and reverse-phase capillary columns separation. The components were separated well by this system and yielded over 420 peaks. Under the optimal condition, 4 compounds were detected quantitatively. A good linear relationship was obtained from 0.2μgmL -1 to 24μgmL -1with detection limits (S/N=3) ranging from 0.05μgmL -1 to 0.2μgmL -1for the compounds. We demonstrated that the method can be successfully applied to the analysis of a natural complex sample, with satisfactory results. © Taylor & Francis Group, LLC.