Relaxin/insulin-like family peptide receptor 4 (RXFP4) is a class A G protein-coupled receptor (GPCR), and insulin-like peptide 5 (INSL5) is its endogenous ligand. Although the precise physiological role of INSL5/RXFP4 remains elusive, a number of studies have suggested it to be a potential therapeutic target for obesity and other metabolic disorders. Since selective agonists of RXFP4 are scarcely available and peptidic analogs of INSL5 are hard to make, we conducted a high-throughput screening campaign against 52,000 synthetic and natural compounds targeting RXFP4. Of the 109 initial hits discovered, only 3 compounds were confirmed in secondary screening, with JK0621-D008 displaying the best agonism at human RXFP4. Its S-configuration stereoisomer (JK1) was subsequently isolated and validated by a series of bioassays, demonstrating a consistent agonistic effect in cells overexpressing RXFP4. This scaffold may provide a valuable tool to further explore the biological functions of RXFP4.
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We are indebted to Ji Wu and Qiang Shen for technical assistance. This work was partially supported by grants from the National Natural Science Foundation of China 81872915 (MWW), 81573479 (DHY), and 81773792 (DHY); the National Science & Technology Major Project “Key New Drug Creation and Manufacturing Program” of China (2018ZX09735-001 to MWW, 2018ZX09711002-002-005 to DHY and 2018ZX09711002-002-011 to QL); the National Key R&D Program of China 2018YFA0507000 (MWW); and the Novo Nordisk-CAS Research Fund (NNCAS-2017-1-CC to DHY). Research at the Florey was supported by the Victorian Government’s Operational Infrastructure Support Program. We are grateful to Tania Ferraro and Sharon Layfield for assistance with assays at the Florey. RADB is an NHMRC Senior Research Fellow. The funders had no role in the study design, data collection and analysis, decision to publish, or paper preparation.
The authors declare no competing interests.
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Lin, Gy., Lin, L., Cai, Xq. et al. High-throughput screening campaign identifies a small molecule agonist of the relaxin family peptide receptor 4. Acta Pharmacol Sin 41, 1328–1336 (2020). https://doi.org/10.1038/s41401-020-0390-x
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