Abstract
Estrogen is central to many physiological processes throughout the human body. We have previously shown that the G protein–coupled receptor GPR30 (also known as GPER), in addition to classical nuclear estrogen receptors (ERα and ERβ), activates cellular signaling pathways in response to estrogen. In order to distinguish between the actions of classical estrogen receptors and GPR30, we have previously characterized G-1 (1), a selective agonist of GPR30. To complement the pharmacological properties of G-1, we sought to identify an antagonist of GPR30 that displays similar selectivity against the classical estrogen receptors. Here we describe the identification and characterization of G15 (2), a G-1 analog that binds to GPR30 with high affinity and acts as an antagonist of estrogen signaling through GPR30. In vivo administration of G15 revealed that GPR30 contributes to both uterine and neurological responses initiated by estrogen. The identification of this antagonist will accelerate the evaluation of the roles of GPR30 in human physiology.
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Acknowledgements
This work was supported by NIH grants CA118743 and CA127731 (to E.R.P.), by grants from the Oxnard and Stranahan Foundations (to E.R.P.), by the University of New Mexico Center for Molecular Discovery (NIH U54 MH084690; to L.A.S.) by the New Mexico Tobacco Settlement fund (to T.I.O.), by the New Mexico Cowboys for Cancer Research (to J.B.A.) and by NIH grants R37 NS18710 (to N.J.D.) and HL90804 (to E.B.). Flow cytometry data and confocal images in this study were generated in the Flow Cytometry and Fluorescence Microscopy Facilities, which received support from the University of New Mexico Health Sciences Center and the University of New Mexico Cancer Center (as detailed at http://hsc.unm.edu/crtc/microscopy/facility.html). In vivo data were generated with the support of the University of New Mexico Cancer Center Animal Models & Imaging Core.
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M.K.D. designed, performed and analyzed biomolecular screening, ligand binding and cell biology experiments. W.K.P., S.N.A. and H.J.H. designed, performed and analyzed animal uterine experiments. T.K.N. designed, performed and analyzed ligand binding experiments. E.B., E.D. and N.J.D. designed, performed and analyzed animal depression experiments. C.G.B., A.L. and T.I.O. designed, performed and analyzed virtual screening experiments. R.B., C.R. and J.B.A. synthesized G-1 and G15. L.A.S. provided integration with the University of New Mexico MLSC. E.R.P. designed and analyzed experiments and, with contributions from all authors, wrote the paper.
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Dennis, M., Burai, R., Ramesh, C. et al. In vivo effects of a GPR30 antagonist. Nat Chem Biol 5, 421–427 (2009). https://doi.org/10.1038/nchembio.168
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DOI: https://doi.org/10.1038/nchembio.168
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