Design,Synthesis And Antitumor Activity Of Indole-Pyrimidine Derivatives As Nuclear Receptor Modulators

Fragment-based drug design and molecular hybridization are two classic methods of drug design,which play an important role in the development of modern drugs,and pyrimidine and indole are widely used as novel fragments for the development of new drugs.In addition,nuclear receptors and kinases play important roles in the occurrence and development of various diseases,such as tumors,inflammation,and metabolism.Therefore,the main work of this paper is to design and synthesize targets that target these targets based on the structural characteristics of the binding sites of the targets(Nur77,RXR?,PPAR?,CDK9,HDAC),using molecular hybridization technology and fragment-based drug design strategies.Pyrimidine-indole derivatives and preliminary research on their biological activities.Research on novel pyrimidine-indole Nur77 agonists:Based on the Nur77 agonists that have been discovered,based on molecular hybridization technology and fragment-based drug design methods,122 pyrimidine-indole derivatives with a new mother nucleus were designed and synthesized.Reporter experiments and Biacore test results show that most of the compounds have transcriptional activation activity against Nur77 and can bind to Nur77-LBD protein.Among them,QH117 was able to selectively activate Nur77 transcriptional activation activity,and Kd=91nM bound to protein Nur77.Further biological mechanism studies have shown that the compound QH117 can promote the apoptosis of MDA-MB-231 cells in a time-and concentration-dependent manner.The apoptosis was determined by Nur77 by Western blot and cell flow experiments.Computer simulations and amino acid mutation experiments proved that compound QH117 can bind to key sites such as GLU445,ARG515,GLN528 of Nur77 protein.Fluorescence immunoassay confirmed that QH117 can promote the cleavage of Cas 3.Finally,mouse transplantation tumor experiments show that compound QH117 can inhibit the growth of MDA-MB-231,and at the same time exhibits characteristics of high efficiency and low toxicity.QH117 can be further developed as a seed compound.Study on novel pyrimidohydrazide urea RXR? antagonists:Through molecular hybridization strategies and fragment-based drug design methods,pyrimidohydrazide urea RXR? antagonists with a new structure type were designed and 50 compounds were synthesized.The structure-activity relationship analysis of pyrimidine hydrazide urea compounds was performed by the reporter gene and SPR combined with small molecules to inhibit tumor cell proliferation experiments.Finally,compound 6A was found to have high binding activity to RXR?,Kd=120 nM,which is a selective RXR? antagonist Agent,can inhibit tumor cell proliferation well,and is almost non-toxic to normal cells.Further biological mechanism studies show that 6A can rely on RXR? to induce apoptosis,and has better anti-tumor proliferative activity than anti-tumor drugs in terms of anti-tumor,which deserves further research.Research on novel 1,3,5 substituted indolepropionic acid PPAR? inhibitors:By analyzing the published small molecule and PPAR? protein crystal binding mode,combined with molecular hybridization technology and computer drug-assisted design methods,we designed and synthesized a novel 1 108,3,5 substituted indolepropionic acid PPAR? inhibitors,and their effects on PPAR?-LBD transcriptional activity were determined by reporter genes.It was found that most compounds can inhibit PPAR?-LBD transcriptional activation activity and can selectively inhibit PPAR?-LBD transcription activation activity.The Biacore method was used to determine the binding activity of the compounds to the PPAR?-LBD protein,and many of them were found to be able to bind to the PPAR?-LBD protein.The Kd of QL3,QL6,QL9,and QL11 compounds bound to the protein reached nanomolar levels.In the later stage,we will analyze the structure-activity relationship of the compounds based on the current biological data and conduct further research on their inhibitory activity on tumor proliferation.Research on novel CDK-HDAC dual target inhibitors:By analyzing the signaling pathways of CDK and HDAC in tumors,we find that they have synergistic effects on cycle block and can promote tumor proliferation.According to the multi-target drug design strategy and based on the Fragment Drug Design(FBDD)technology,we designed and synthesized a series of new pyrimidine-indole derivatives.Among them,compound 13ea was found in several cancer cell lines(HeLa,MDA-MB-231).,HepG2)all showed strong and effective antiproliferative activity.Compound 13ea can inhibit both CDK9(IC50=166nM)and HDAC activity(IC50=NA).At a low concentration(0.625 ?M),the cells undergo cell cycle arrest and induce apoptosis,and the effect of inducing apoptosis is stronger than that of positive drugs(BG45,CI-994,Entinostat and Mocetinostat).Finally,in vivo experiments have shown that 13ea can significantly inhibit the growth of MDA-MB-231 tumors,and docking studies have also shown that 13ea can bind CDK9 and HDAC2 well.This research provides a certain theoretical and experimental basis for the further development of a dual-target anti-cancer drug of CDK-HDAC.