作者: Wurnig, Silas L. ; Kostenis, Evi ; Toy, Lara ; Alonso Bartolomé, Ana ; Günther, Stefan ; Schiedel, Matthias ; Moumbock, Aurélien F. A. ; Huber, Max E. ; Szpakowska, Martyna ; Hansen, Finn K. ; Chevigné, Andy

AbstractA conserved intracellular allosteric binding site (IABS) was recently identified at several G protein‐coupled receptors (GPCRs). This target site allows the binding of allosteric modulators and enables a new mode of GPCR inhibition. Herein, we report the development of a NanoBRET‐based assay platform based on the fluorescent ligand LT221 (5), to detect intracellular binding to CCR6 and CXCR1, two chemokine receptors that have been pursued as promising drug targets in inflammation and immuno‐oncology. Our assay platform enables cell‐free as well as cellular NanoBRET‐based binding studies in a nonisotopic and straightforward manner. By combining this screening platform with a previously reported CXCR2 assay, we investigated CXCR1/CXCR2/CCR6 selectivity profiles for both known and novel squaramide analogues derived from navarixin, a known intracellular CXCR1/CXCR2 antagonist and phase II clinical candidate for the treatment of pulmonary diseases. By means of these studies we identified compound 10, a previously reported tert‐butyl analogue of navarixin, as a low nanomolar intracellular CCR6 antagonist. Further, our assay platform clearly indicated intracellular binding of the CCR6 antagonist PF‐07054894, currently evaluated in phase I clinical trials for the treatment of ulcerative colitis, thereby providing profound evidence for the existence and the pharmacological relevance of a druggable IABS at CCR6.

Nature Communications

Structural basis for CCR6 modulation by allosteric antagonists

Article

作者: Li, Wei ; Mondal, Prolay ; Lombardo, Vincent ; Calabrese, Matthew F. ; Wu, Huixian ; Unwalla, Ray J. ; Hogan, Barbara J. ; Cooke, Robert M. ; Frisbie, Richard K ; Vincent, Fabien ; Alley, Jennifer ; Gerstenberger, Brian S ; Thorarensen, Atli ; Mousseau, James J. ; Farley, Kathleen A ; Segala, Elena ; Ding, Ru ; Berstein, Gabriel ; Rappas, Mathieu ; Chinigo, Gary ; Schnute, Mark E ; Oswald, Christine ; Minhas, Gurdeep Singh ; Hogan, Barbara J ; Trujillo, John I. ; Nuhant, Philippe ; Han, Seungil ; Frisbie, Richard K. ; Crouse, Kimberly K. ; Victorino, Jorge ; Dermenci, Alpay ; Unwalla, Ray J ; Calabrese, Matthew F ; Flick, Andrew ; Crouse, Kimberly K ; Farley, Kathleen A. ; Gerstenberger, Brian S. ; Choi, Chulho ; Trujillo, John I ; Sahasrabudhe, Parag V ; Schnute, Mark E. ; Wasilko, David Jonathan ; Windsor, Ian W. ; Mousseau, James J ; Lamb, Daniel ; Windsor, Ian W ; Cooke, Robert M ; Sahasrabudhe, Parag V.

The CC chemokine receptor 6 (CCR6) is a potential target for chronic inflammatory diseases. Previously, we reported an active CCR6 structure in complex with its cognate chemokine CCL20, revealing the molecular basis of CCR6 activation. Here, we present two inactive CCR6 structures in ternary complexes with different allosteric antagonists, CCR6/SQA1/OXM1 and CCR6/SQA1/OXM2. The oxomorpholine analogues, OXM1 and OXM2 are highly selective CCR6 antagonists which bind to an extracellular pocket and disrupt the receptor activation network. An energetically favoured U-shaped conformation in solution that resembles the bound form is observed for the active analogues. SQA1 is a squaramide derivative with close-in analogues reported as antagonists of chemokine receptors including CCR6. SQA1 binds to an intracellular pocket which overlaps with the G protein site, stabilizing a closed pocket that is a hallmark of inactive GPCRs. Minimal communication between the two allosteric pockets is observed, in contrast to the prevalent allosteric cooperativity model of GPCRs. This work highlights the versatility of GPCR antagonism by small molecules, complementing previous knowledge of CCR6 activation, and sheds light on drug discovery targeting CCR6.

100 项与 CCR6调节剂(Idorsia) 相关的药物交易

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