DNA-Encoded Library Screening Identifies Benzo[b][1,4]oxazepin-4-ones as Highly Potent and Monoselective Receptor Interacting Protein 1 Kinase Inhibitors

Abstract

The recent discovery of the role of receptor interacting protein 1 (RIP1) kinase in tumor necrosis factor (TNF)-mediated inflammation has led to its emergence as a highly promising target for the treatment of multiple inflammatory diseases. We screened RIP1 against GSK's DNA-encoded small-molecule libraries and identified a novel highly potent benzoxazepinone inhibitor series. We demonstrate that this template possesses complete monokinase selectivity for RIP1 plus unique species selectivity for primate versus nonprimate RIP1. We elucidate the conformation of RIP1 bound to this benzoxazepinone inhibitor driving its high kinase selectivity and design specific mutations in murine RIP1 to restore potency to levels similar to primate RIP1. This series differentiates itself from known RIP1 inhibitors in combining high potency and kinase selectivity with good pharmacokinetic profiles in rodents. The favorable developability profile of this benzoxazepinone template, as exemplified by compound 14 (GSK'481), makes it an excellent starting point for further optimization into a RIP1 clinical candidate.

RIPK3 Activates Parallel Pathways of MLKL-Driven Necroptosis and FADD-Mediated Apoptosis to Protect against Influenza A Virus

“…Next, we employed kinase inhibitors specific for either RIPK1 [GSK’481 (Harris et al, 2016), GSK’963 (Berger et al, 2015), and Necrostatin-1 (Degterev et al, 2008)] or RIPK3 [GSK’872 and GSK’843 (Mandal et al, 2014)] to identify roles for the kinase activity of each of these proteins in IAV-induced RIPK3-RIPK1 complex assembly. Downstream of TNFR1, the kinase activity of RIPK1, but not RIPK3, was necessary for necrosome formation (Fig.…”

CETSA quantitatively verifies in vivo target engagement of novel RIPK1 inhibitors in various biospecimens

“…Several types of RIPK1 inhibitors have been reported 16,22,27–30 . We validated our assay by investigating the dose response curves of a total of 14 compounds: Nec-1, GSK-compound 27 , and twelve 7-oxo-2,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine derivatives (Fig.…”

DNA‐encoded chemical libraries – achievements and remaining challenges

“…This compound was derived from a candidate (IC 50 = 10 nM) (Table 1 (entry d)) that originated from a split-andpool DECL library with three sets of building blocks, two of which proved to be essential for RIP1 binding [57].…”