Design,Synthesis And Antitumor Activity Of Novel Skp2 Inhibitors

Small molecule chemical therapy is one of the important means to treat malignant tumors.Finding and developing new small molecule anti-tumor drugs is an important measure for the treatment of malignant tumors.Numerous studies have shown that S-phase kinase-associated protein?Skp2?is a recently reported novel gene that is highly expressed in many malignant tumors and has proto-oncogene function.It can promote the proliferation of cancer cells by and regulating the activity of Cul1-Rbx1-Skp1-F-box^Skp2 complex to inhibit p27 gene and promote glycolysis in cancer cells.Recent reports indicate that Skp2 inhibitors inhibit the function of cancer stem cells and control the progression of cancer by activating p27 and inhibiting cell glycolysis.In summary,Skp2 is a promising target in the field of cancer therapy,and the study of Skp2 inhibitors and further research on its mechanism of action are of great significance.The aim of this study was to design and synthesize inhibitors that target the interaction between Skp2-Skp1 proteins,which laid the foundation for further research and development of new anti-tumor drugs targeting Skp2.First,by using Discovery Studio 3.1,the characteristic analysis of the binding pocket of Skp2-Skp1 was carried out and the characteristic structure of the reported Skp2 inhibitor SZL-P1-41 was analyzed,and then the zinc database with 300 thousand compounds was screened,and the first 1%of the compounds were selected for molecular docking.The first 300compounds were selected for ADMET pre-assessment and MM/GBSA re-scoring.The first 30 compounds were manually observed for docking.The correct mode of action and the novel core structure was selected.Finally,according to the results of computer-aided drug design screening,the methylene hydrazide-based compounds were selected for further study based on the novelty of the structure and the convenience of synthesis.The core structure of the methylene hydrazide was kept unchanged.According to the pharmacophore characteristics of the protein binding site and the Skp2inhibitor SZL-P1-41,dozens of different branches were designed,and virtual combination of branch and core structures to form a new compound library.Then molecular docking with Skp2,pre-evaluation of ADMET properties and re-evaluation of MM/GBSA binding energy,and finally screening about 60 compounds for chemical synthesis.Finally,about 60 compounds are selected for chemical synthesis.Through A series of chemical synthesis,such as substitution of aromatic hydrazide and Schiff base reaction,HRMS and ¹H-NMR characterization of its structure,49 compounds were finally obtained,and SZL-P1-41 was synthesized as a positive control for in vitro activity screening.Subsequently,7 tumor cell lines were selected and 49 synthetic compounds and SZL-P1-41 were screened for in vitro tumor activity.Finally,more than ten compounds with nanomolar activity are obtained.Among them,compound 15k had an inhibitory activity of 50 nM in HGC27 cells and has good inhibitory activity against all tumor cells.It is predicted that compound 15k is highly likely to be an excellent lead compound with broad spectrum antitumor activity.