Novel Gpr39 Agonists: Correlation Of Binding Affinity Using Label-Free Back-Scattering Interferometry With Potency In Functional Assays
Poster Sep 02, 2014
Daniel Brown (1), Niklas Larsson (2), Ola Fjellström (3), Anders Johansson (3), Sara Lundqvist (2), Johan Brengdahl (2), and Richard J. Isaacs (1)
Back-scattering interferometry (BSI) is an emerging label-free, conformation-sensitive detection technology for quantitative mass- and matrix-independent biophysical characterization of small molecule interaction with complex drug target proteins under native-like conditions (1). Integral membrane proteins such as GPCRs are critical targets for drug discovery but present a host of challenges to the investigation of their biophysical properties. Of paramount interest to drug discovery efforts is the characterization of the interaction of GPCRs with small molecule compounds as a component of library screening, mechanism of action (MOA) determination, drug candidate profiling, and other aspects of intermolecular binding that inform pharmacology and medicinal chemistry. The difficulty associated with obtaining small molecule affinity data for functionally intact GPCRs effectively restricts the range of assay techniques suited to quantifying these interactions in vitro.
Herein, we describe the application of BSI to the characterization of small molecule ligand binding to human GPR39 overexpressed in crude membrane fractions in free solution. GPR39 is a Zn2+-responsive GPCR under investigation as a therapeutic target for type-2 diabetes (2). The ability to measure the affinity of small molecule agonists such as Zn2+is especially novel, given the unfavorable mass ratio and fast off rate that complicates the use of more established binding assays. Results from screening representatives from multiple novel GPR39 agonist series is presented, including how BSI-derived affinity and functional assay-derived potency correlate for compounds of varying scaffolds.
P450 Induction in Cryopreserved Hepatocytes from PXR and CAR Nuclear Receptor Knock-out RatsPoster
The nuclear receptors pregnane X receptor (PXR) and constitutive androstane receptor (CAR) are closely related transcription factors that regulate the expression of phase I (cytochrome P450s), phase II metabolizing enzymes and transporter genes in response to xenobiotics, including prescription drugs.READ MORE
Regulatory T-Cells (Tregs) Within Bone Marrow-Derived Stem Cells (BMSCs) Actively Confer Immunomodulatory and Neuroprotective Effects Against StrokePoster
We found a distinct subpopulation of Tregs within BMSCs. Tregs and BMSCs in co-culture conferred neuroprotection that varied in a dose-dependent manner. Tregs minimized stem cell production of IL-6, a pro-inflammatory cytokine, and inhibited BMSC secretion of FGF-beta, a cytokine related to BMSC proliferation and differentiation. The ratio of Tregs found natively in BMSCs is optimally adapted to provide the maximum neuroprotective benefit of stem cell treatment after ischemic stroke.READ MORE
Internet-Based Biomarker Collection Feasibility: Experiences of a Tobacco Cessation Program for People Living with HIVPoster
This project introduces video-conferencing as a method for biomarker collection in research and will explain how biomarker collection via video-conferencing was implemented in a pilot ehealth intervention.READ MORE