Discovery and characterization of highly potent and selective allosteric USP7 inhibitors

Abstract

Given the importance of ubiquitin-specific protease 7 (USP7) in oncogenic pathways, identification of USP7 inhibitors has attracted considerable interest. Despite substantial efforts, however, the development of validated deubiquitinase (DUB) inhibitors that exhibit drug-like properties and a well-defined mechanism of action has proven particularly challenging. In this article, we describe the identification, optimization and detailed characterization of highly potent (IC < 10 nM), selective USP7 inhibitors together with their less active, enantiomeric counterparts. We also disclose, for the first time, co-crystal structures of a human DUB enzyme complexed with small-molecule inhibitors, which reveal a previously undisclosed allosteric binding site. Finally, we report the identification of cancer cell lines hypersensitive to USP7 inhibition (EC < 30 nM) and demonstrate equal or superior activity in these cell models compared to clinically relevant MDM2 antagonists. Overall, these findings demonstrate the tractability and druggability of DUBs, and provide important tools for additional target validation studies.

Targeting the overexpressed USP7 inhibits esophageal squamous cell carcinoma cell growth by inducing NOXA‐mediated apoptosis

“…A serious of inhibitors targeting USP7 was evaluated in vitro and in vivo studies . In which, P5091 was identified as a specific inhibitor of USP7 and showed a promising anti‐cancer effect in several tumors .…”

“…Several inhibitors targeting USP7/HAUSP were screened and exhibited anti‐tumor effect . Chauhan et al discovered that P5091 exhibited specific and selective deubiquitylating activity against USP7 and did not inhibit other DUBs.…”

Palladium‐Mediated Cleavage of Proteins with Thiazolidine‐Modified Backbone in Live Cells

“…[48][49][50] One interesting example was reported by Ernst et al [51] where directed evolution was used to generate ar ange of Ub variants (Ubvs) that selectively target their corresponding DUBs. [48][49][50] One interesting example was reported by Ernst et al [51] where directed evolution was used to generate ar ange of Ub variants (Ubvs) that selectively target their corresponding DUBs.…”

Targeting ubiquitin specific protease 7 in cancer: A deubiquitinase with great prospects

“…Through this platform, specific UbVs have been identified that can inhibit USP7 and USP10 with high affinity . Implementation of fragment‐based methods and surface plasmon resonance (SPR) has led to the generation of new compounds with high inhibition potency . Combination of features of published USP7 inhibitors and binding fragments has resulted in Compound 1 as exemplified by t7‐Bromo‐3‐((4‐hydroxy‐1‐(3‐phenylpropanoyl)piperidin‐4‐yl)methyl)thieno[3,2‐d]pyrimidin‐4(3H)‐one (IC 50 13.1 μM) which is later developed to generate the potent compound 4 (IC 50 6 nM).…”