Targeting Src Structure Of Protein SH3 Domain Antitumor Drugs The Discovery Of Lead Compounds

The c-Src kinase is the maximum studied membership of the Src family(SFK),the largest non-receptor tyrosine kinase family.The overexpression or activation of cSrc is related to the abnormal formation of invasive pseudopodia,which is the key morphological structure for cancer cells to infiltrate into the blood and exudate to the secondary site during metastasis.Therefore,the overexpression and activation of c-Src in malignant tumors are related to the development,progression and metastasis of cancer.At present,there are inhibitors targeting c-Src protein on the market,and these c-Src inhibitors inhibit the activity of c-Src by competing with ATP for the mechanism of binding sites.Proline-rich recognition modules are widely distributed in the SH3 domain(Src Homology3)of c-Src,which can bind to other proteins and themselves,so the SH3 structure is the key structure for substrate recruitment and supramolecular complex formation of c-Src proteins,which regulates the kinase activity of c-Src.Activated cSrc interacts with many downstream targets responsible for tumor tissue occurrence,development,angiogenesis,cancer cell proliferation and movement to exert its physiological function.At present,the listed drugs targeting c-Src are mainly ATP competitive inhibitors acting on the kinase domain of the c-Src domain,but because they target the kinase domain,the kinase domain is prone to allostery under the intermolecular force,resulting in drug mistargeting,while the SH3 domain,as an important part of the kinase function of c-Src,has not been listed as a related targeted anticancer drug.The purpose of this study is to develop new small molecular lead compounds targeting the SH3 domain of c-Src protein to solve the problem of drug resistance of competitive inhibitors of ATP.Therefore,we first obtained the constructed plasmid expressing c-Src SH3 protein and purified it to obtain high-purity c-Src SH3 domain protein.The compound in the compound library was bound and screened for kinase activity at the molecular level by protein thermal migration experiment and kinase experiment respectively,and then the proliferative inhibitory activity was screened at the cell level.The characterization of the compound on the cell was further studied by simulated cell environment experiment,migration experiment,invasion experiment,dry pellet formation experiment,cycle experiment and apoptosis experiment.Finally,the targeting of c-Src in cells was verified by Westernblot assay,and the effect of these compounds on cell function was preliminatively explored.In this study,three new anti-tumor lead compounds targeting SH3 were found,and related cell characterization experiments and preliminary mechanism studies were conducted around them,which laid an important foundation for subsequent anti-tumor studies targeting c-Src SH3 and provided a new idea for clinical anti-tumor drug resistance.