This paid-for advertorial by Eurofins Discovery appeared in Volume 23 – Issue 3, Summer 2022.
In the following article, Luke Oostdyk, Lakshmi Anantharaman and Nicole Servant from Eurofins Discovery discuss how G-Protein Coupled Receptor screening and profiling can advance drug discovery programs.
The sheer number of understudied and orphan receptors in the G-Protein Coupled Receptor (GPCR)1 family have made them such an attractive target for drug discovery programs that one-third of all FDA-approved drugs target them. The Eurofins Discovery GPCR screening and profiling services offer a suite of unique and robust cell-based assays to screen for agonists, antagonist and allosteric modulators for the majority of the GPCR family.
GPCR Panel Offerings Support Target Identification for Compounds of Interest
Figure 1: gpcrMAX and orphanMAX Panels. A. gpcrMAX panel agonist screening FDA-approved UK14304 (α2 adrenergic agonist) and endogenous neurotransmitters acetylcholine and serotonin, and antagonist screening for FDA-approved antagonists Naloxone (opioid), Betaxolol (beta-adrenergic) and Prazosin (alpha-adrenergic). B. Dose response curve hit confirmation testing in both β-arrestin and second messenger (cAMP and Ca¬++) assays for acetylcholine agai nst the muscarinic receptor family. C. orphanMAX screening of the bacterial effector Commendamide, and the successful confirmation of activity in GPR132 (inset). D. Metrics for the gpcrMAX and orph anMAX panels.To aid determining the mechanism-of-action of bioactive molecules in drug discovery programs, Eurofins Discovery offers GPCR panels for both liganded (gpcrMAX)and non-liganded orphan (orphanMAX) receptors that utilize β-arrestin recruitment, to rapidly, reliably and reproducibly profile compounds using a common G-protein independent readout. Panels are suited for primary screening of small molecules, peptides or biologics, and offered on regular 6-week schedules.
The gpcrMAX panel contains a set of 168 GPCRs covering over 60 distinct receptor families that demonstrate specificity for FDA-approved drugs and endogenous signalling molecules in both agonist and antagonist modes (Figure 1A). Eurofins Discovery’s vast cell-based assay catalogue enables follow-up specificity and potency testing in both β-arrestin and second messenger read-outs (Figure 1B).
The orphanMAX panel includes a collection of 73 human orphan GPCRs capable of uncovering novel ligand and receptor pairings, offering the ideal solution for any ligand fishing or deorphanisation campaign. The Eurofins Discovery gpcrMAX and orphanMAX panels offer simultaneous screening against 241 GPCRs with 409 total assays (Figure 1D).
As a testament to the panels’ potential effectiveness, Eurofins Discovery supported bioscience researchers at the Rockefeller University (USA) using these panels in screening bacterial effectors from the human microbiome: An initial hit in the orphanMAX panel was confirmed with follow-up dose response curve testing and identified GPR132 as the target of an N-acyl-amide given the name Commendamide(Figure 1C).2
Unmatched Speed in GPCR High-Throughput Screening and Lead Optimization
Eurofins Discovery combines rapid pace, portfolio of β-arrestin and second messenger assays, and technical expertise to provide end-to-end solutions for high-throughput screening (HTS), and hit-to-lead and lead optimization programs.
A recent example: Eurofins Discovery partnered with a GPCR-focused pharmaceutical client screening against eight assays in an emerging GPCR subfamily. Six of the eight assays were available for immediate use, allowing screening to begin while development of the other two was performed. The client’s 100,000-compound library was screened in just 6 days per target (Table 1A) with an average hit rate of 0.43%(430 hits/100k screen) and an excellent Z ́ average (0.59). The result: Completion of all six screens in 6 weeks – half the turnaround time compared to~90 days at other CROs.
For critical lead optimization Eurofins Discovery continued to support its client; weekly screening and 5-day turnaround times supplied the data necessary to inform the client’s chemist syntheses and minimize delays during new molecule development(Table 1B).
References
1. Hauser AS, Attwood M., Rask-Andersen M, Schiöth HB, Gloriam DE. Trends in GPCR drug discovery: new agents, targets and indications. Nature Reviews Drug Discovery. 2017;16(12), pp.829-842.
2. Cohen LJ, Kang HS, Chu J, HuangYH, Gordon EA, Reddy BVB, Ternei MA, Craig JW, Brady SF. Functional metagenomic discovery of bacterial effectors in the human microbiome and isolation of commendamide,a GPCR G2A/132 agonist. Proceedings of the NationalAcademy of Sciences USA.2015;112(35), pp.E4825-E4834.
