Design,Synthesis And Anti-Tumor Activity Evaluation Of Novel 3,5-Diarylpyridine USP7 Inhibitors

Ubiquitin specific protease 7(USP7/HAUSP)is an important epigenetic modification protein.It can specifically hydrolyze ubiquitin molecules from proteins or precursor proteins and exert reverse regulation of proteins degradation to keep the substrate proteins stable,thus affecting their functions.Studies have shown that USP7 is highly expressed and abnormally activated in a variety of tumor cells,and it is closely related to DNA damage,apoptosis and cell cycle;Inhibition of its activity or reduction of its expression can effectively inhibit the occurrence and development of tumors.Anti-tumor drugs targeting USP7 have become a research hotspot and received more and more attention.However,existing USP7 inhibitors can exhibit lack of structural types,poor selectivity,and low specificity,thus leading to the development of USP7 inhibitors with high efficiency,low toxicity and high selectivity becoming the focus of human attention.As an important heterocyclic pharmacophore,pyridine is widely used in the fields of medicine,materials,pesticides,perfumes and dyes.Derivatives with pyridine as the skeleton are also widely used in the treatment of various diseases.Inspired by this,this article focused on the structural optimization of the reported pyridine derivatives,a series of 3,5-diaryl pyridine derivatives were designed and synthesized,and their enzymatic activities and primary biological mechanisms were investigated.In detailed studies are as follows:First,By referring to relevant references,we had a preliminary understanding of the protein structure,biological function and the role of USP7 in the development of cancer.By referring to relevant references,we had a preliminary understanding of the protein structure,biological function and the role of USP7 in the development of cancer.The research progress of USP7 and the challenges that need to be overcame have been summarized,which provided a theoretical basis for the smooth development of this topic.Second,based on previous research,we found that 3,5-diarylpyridine derivatives can inhibit USP7 activity by binding to the relatively conserved active site 2 of USP7.At the same time,studies have confirmed that the amino acid residues Gln351 of USP7 active site 1 is crucial.By interacting with Gln351,the inhibitory activity of small molecule against USP7 could be improved effectively.We used a benzyl group as the“Linker” on the basis of 3,5-diarylpyridine USP7 inhibitors which acted on active site2.A total 63 novel pyridine derivatives were designed and synthesized by introducing hydrophilic groups as the hydrogen-bond donor and acceptor into the para-position of benzyl Linker,such as amide and hydroxamic acid.It was expected to not only maintain the original binding interaction with the active site 2 of USP7,but also interact with the key residue of Gln351 in the active site 1,thereby improving the inhibitory activity of USP7.Third,The above synthesized compounds were evaluated for their USP7 inhibitory activity.The results showed that compared with the lead compound,the inhibition effect on USP7 of these compounds were significantly increased due to the introduction of the extended side chain,which preliminarily verified the rationality of our design.Besides,given the research that USP7 can promote the development of cervical cancer,we used HeLa as the test cells to evaluate the proliferation inhibitory activity of the representative compounds.Considering the good enzymatic potency and cell proliferation inhibitory activity,compound 52 was selected to study its preliminary mechanism.The results showed that compound 52 could effectively inhibit the migration of HeLa cells and promote their apoptosis.In summary,with the help of a structure-based drug design approach,we performed a lead optimization on the basis of 3,5-diarylpyridine-based USP7 inhibitors that acted on the active site 2.In summary,with the help of a structure-based drug design approach,we performed a lead optimization on the basis of 3,5-diarylpyridinebased USP7 inhibitors that acted on the active site 2.A total of 63 novel pyridines derivatives were designed and synthesized to enrich the structural types of USP7 inhibitors.Anti-migration and apoptosis induced by representative compound 52 in HeLa cells also provided a data support for the discovery of anti-cervical cancer drugs.