title: High throughput screening for orphan and liganded GPCRs.
abstract: GPCRs had significant representation in the drug discovery portfolios of most major commercial drug discovery organizations for many years. This is due in part to the diverse biological roles mediated by GPCRs as a class, as well as the empirical discovery that they have proven relatively tractable to the development of small molecule therapeutics. Publication of the human genome sequence in 2001 confirmed GPCRs as the largest single gene superfamily with more than 700 members, furthering the already strong appeal of addressing this target class using efficient and highly parallelized platform approaches. The GPCR research platform implemented at Amgen is used as a case study to review the evolution and implementation of available assays and technologies applicable to GPCR drug discovery. The strengths, weaknesses, and applications of assay technologies applicable to G alpha s, G alpha i and G alpha q-coupled receptors are described and their relative merits evaluated. Particular consideration is made of the role and practice of "de-orphaning" and signaling pathway characterization as a pre-requisite to establishing effective screens. In silico and in vitro methodology developed for rapid, parallel high throughput hit characterization and prioritization is also discussed extensively.
authors.affiliation: Chemistry Research and Discovery, Amgen Inc., 1120 Veterans Blvd., South San Francisco, CA 94080, USA. Josh.Xiao@Amgen.com
authors.forename: Shou-Hua
authors.initials: SH
authors.lastname: Xiao
authors.forename: Jeff D
authors.initials: JD
authors.lastname: Reagan
authors.forename: Paul H
authors.initials: PH
authors.lastname: Lee
authors.forename: Angela
authors.initials: A
authors.lastname: Fu
authors.forename: Ralf
authors.initials: R
authors.lastname: Schwandner
authors.forename: Xiaoning
authors.initials: X
authors.lastname: Zhao
authors.forename: Johannes
authors.initials: J
authors.lastname: Knop
authors.forename: Holger
authors.initials: H
authors.lastname: Beckmann
authors.forename: Stephen W
authors.initials: SW
authors.lastname: Young
mesh_terms: D000818:Animals; D019390:Arrestins / Q000032:analysis; D020013:Calcium Signaling / Q000187:drug effects; D000242:Cyclic AMP / Q000032:analysis; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006801:Humans; D008024:Ligands; D043562:Receptors, G-Protein-Coupled / Q000819:agonists* / Q000037:antagonists & inhibitors* / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D054852:Small Molecule Libraries / Q000494:pharmacology; D000071557:beta-Arrestins
chemical_list: D019390:Arrestins; D008024:Ligands; D043562:Receptors, G-Protein-Coupled; D054852:Small Molecule Libraries; D000071557:beta-Arrestins; D000242:Cyclic AMP