Discovery of the First Selective Nanomolar Inhibitors of ERAP2 by Kinetic Target‐Guided Synthesis

Abstract

Endoplasmic reticulum aminopeptidase 2 (ERAP2) is a key enzyme involved in the trimming of antigenic peptides presented by Major Histocompatibility Complex class I. It is a target of growing interest for the treatment of autoimmune diseases and in cancer immunotherapy. However, the discovery of potent and selective ERAP2 inhibitors is highly challenging. Herein, we have used kinetic target-guided synthesis (KTGS) to identify such inhibitors. Cocrystallization experiments revealed the binding mode of three different inhibitors with increasing potency and selectivity over related enzymes. Selected analogues engage ERAP2 in cells and inhibit antigen presentation in a cellular context. 4 d (BDM88951) displays favorable in vitro ADME properties and in vivo exposure. In summary, KTGS allowed the discovery of the first nanomolar and selective highly promising ERAP2 inhibitors that pave the way of the exploration of the biological roles of this enzyme and provide lead compounds for drug discovery efforts.

Alternative drug discovery platforms for the identification of anti-infective agents

“…to discover small molecule binders for numerous biological targets including acetylcholine esterase, [11][12] DNA, 13,14 carbonic anhydrase II, [15][16] , HIV-1 protease, 17 Bcl-XL, 18,19,20 Serratia marcescense chitinase, 21 human sirtuin 1, 22 mycobacterium tuberculosis EthR, 23 acetylcholine binding protein, 24 Bcr-Abl, 25 NAD Kinase, 26 biotin protein ligase, 27 14-3-3 protein, 28,29 insulindegrading enzyme, 30 bacterial ribosome, 31,32 protein factor Xa, 33 cyclooxygenase-2, 34 STAT5, 35 MDM2, 36 endothiapepsin, 37 and ERAP2. 38 KTGS's versatility is highlighted by reports describing some of the technique's most unique applications such as the identification of protein-protein interaction inhibitors, 18,28,29,39,40 discovery of suitable PET imaging probes for human carbonic anhydrase II, 41 assembly of inhibitors on c-MYC G-quadruplex DNA embedded on the surface of gold-coated magnetic nanoparticles, 14 implementation of KTGS in cells, 16,32 and the development of multi-component KTGS methods. 35,37,42 The vast majority of published KTGS literature employs the Huisgen 1,3-dipolar cycloaddition using alkynes and azides to form 1,2,3-triazoles.…”