Synthesis,immunosuppressive Activity And Structure-activity Relationship Study Of The Isoxazole Amide Derivatives

Immunosuppressive agents can inhibit the body's abnormal immune response,it plays an important role in the treatment of autoimmune diseases and organ transplantation anti-rejection.In recent years,more and more new immunosuppressive agents have been developed and used in clinical treatment,although the development of new immunosuppressive agents has made great progress,there are also have some challenges,new immunosuppressive agents research direction is still aimed at low toxicity,efficiency and low cost.The study found that signal factors associated with lymphocytes play an important role in the conduction of mTOR signaling pathways,such as interleukin IL-2,IL-4 and other extracellular cell growth factor through mTOR pathway transmission can promote lymphocyte proliferation.Immunosuppressive agents,acting on the mTOR target,can inhibit the proliferation of lymphocytes by blocking the conduction of the signal,thus play a role in immunotherapy.Computer-aided drug design(CADD)has the advantages of low time and low cost in the course of drug development targeting mTOR,which make the study of new immunosuppressive agents based on CADD has a bright future.In the article,based on the preliminary research of the laboratory,through the transformation of lead compounds,and then design new structures of isoxazolamide compounds,8 isoxazolamides were successfully synthesized by retrosynthesis.The immunosuppressive activity of the compounds was demonstrated by the vitro bioactivity experiments,a total of 25 compounds(The laboratory early synthesized 17 compounds and the 8 compounds were synthesized in this subject)were used to establish a three-dimensional quantitative structure activity relationship model to systematically analyze the relationship between the structure and activity of the compounds,and the advantages of the compound transformation site will be determined based on three-dimensional contour map.The interaction mode of the best activity compound with mTOR receptor protein was analyzed at the molecular level by molecular docking.Based on the results of these experiments to provide a theoretical basis for the development of new immunosuppressive agents with high selectivity,high activity and independent intellectual property rights.The main content of this research 1).Design of Isoxazole Amide DerivativesSimulating the interaction mode of the lead compound with the Rapamycin target protein(mTOR)receptor based on the molecular docking method,and then analyze the binding model of the compound with the receptor protein,thus the novel structures of isoxazolamide derivatives were designed on the basis of the lead compound.2).Synthesis,purification and structure determination of compoundsCompounds were synthesized by retrosynthesis,references to the literature,8 isooxazole amide derivatives were synthesized by a five-step synthetic route,then purifying the compounds and the Liquid Chromatography-Mass Spectrometry(LC-MS)technology and Nuclear magnetic resonance spectroscopy/carbon spectrum(1H NMR/ 13 C NMR)were used to identify the structure of the compounds.3).Determination the vitro biological activity and fitting out the half maximal inhibitory concentrationThe inhibitory activity of 25 compounds(The laboratory early synthesized 17 compounds and the 8 compounds were synthesized in this subject)against Jurkat cells and mouse spleen lymphocytes were determined by CCK-8 cell activity detection technology,then half maximal inhibitory concentration(IC50)were fitted by SPSS.19 software.4).The structure activity relationship and molecular docking studyThe relationship between chemical structure and biological activity of 25 compounds were analyzed based on the 3D-QSAR(three-dimensional quantitative structure activity relationship)model,through the three-dimensional contour map to determine the advantages of the compound transformation sites.The interaction between the small molecule ligands and receptor protein was analyzed by molecular docking,thus study the binding mode relationship between compound and receptor protein.On this account,it guides the structural modification of the new compounds based on 3D-QSAR and molecular docking,which can provide a scientific basis for the design of a better class of this kind amide compounds.The Research Results 1).Based on the molecular docking to analysis the interaction between the lead compound and the receptor protein,8 new structures were modified and synthesized successfully by retrosynthesis,then confirmed the structure of these compound was consistent with the target product,and the purity is more than 98%.2).Get the inhibitory effects of all compounds on the proliferation of Jurkat cells and the half maximal inhibitory concentration(IC50)of each compound;The inhibitory effects of NO.15 compound(the best active compound)on the proliferation of mouse spleen lymphocytes were determined.3).The 3D-QSAR model was successfully constructed and the relationship between the structure and activity of the compound was analyzed in detail.Through the three-dimensional contour map get the advantages of compound transformation sites,and obtain the binding mode of the compound with receptor protein based on the molecular docking.