| Background: With the development of molecular biology and structural biology,more and more three-dimensional structure of biomacromolecule as recepter was discovered and detected.Structure-based durg design means that using known biological macromolecular structure to select interaction of small molecules with the help of computational chemistry and computational biology.This method can reduce the time and money of the new drug research,which has widely applicated in the field of current drug development.Tumor is a disease that threatens health,it has always been the goal of scientists dedicated to conquer.Cyclin dependent kinase,often called CDKs,is a member of Ser/Thr protein kinase family and mainly involved in cell cycle regulation.CDK2 is one of the subtypes,which can promote cell cycle transition from S-phase to G1 and coordinate cell cycle progression.Researches have shown that overexpression of CDK2 appears in more than 80 percent of cancer,such as liver cancer,breast cancer.So finding CDK2 inhibitors is helpful to find new anti-cancer drugs.Signal transducer and activator of transcription 3,often called STAT3,participate in the JAK-STAT3 signal pathway activated by cytokine stimulation.The signal pathways in the regulation of gene transcription in the nucleus and plays an important role in maintaing normal cell function.However,STAT3 over-activation could induce a series high-leval expression of gene which promote cell proliferation,malignant transformation and inhibite apoptosis.These changes result in cancer occurrence and development,STAT3 has become an important target for finding anti-cancer drugs due to the overexpession of STAT3 in more and more cancers.The development of CDK2-cyclinA and STAT3 three-dimensional structure provides a basis for the specificity of small molecule inhibitors.Objective: We designed the ATP competitive inhibitors of CDK2 and STAT3 inhibitors acting on the SH2 pockets by using CDK2-cyclin A and STAT3 three-dimensional structure.Methods: Based on three-dimensional structure of CDK2-cyclin A and STAT3,we used methed of computer aided drug design to conduct molecular docking in commercial compound library,then scored and sorted.After that,we purchased compound with high scores.For inhibitor of CDK2,we tested biological activity,and then selected the best one to perform chemical modification.After getting a series of small molecules with the same parent nuclear,we tested biological activity and chose the best for CCK8.Then we have verified its competitiveness;For inhibitor of STAT3,we tested biological activity by FP assay and then the small molecules with good activity were subjected to CCK8 experiments in MDA-MB-468 cells.After that,we chose the best for CCK8 in more cancer cells.Then tested acting site by Biacore.Results: CDK2 inhibitors: we got compound 81 from the purchase of small molecules and IC50 was 30.34?M,then we modified the compound using pharmaceutical chemistry and get the ATP competitive inhibitor WZ-26,the activity increased 10 times compared with 81 and IC50 was 3.81?M.Cell proliferation inhibition in HepG2 showed antitumor activity,IC50 was 30.58?M.STAT3 inhibitors: we used FP assay to choose compound and got compound B9.In MDA-MB-468 cells,antiproliferative activity was best and IC50 was 5.1?M,besides,in MDA-MB-231 and DU145 cells,B9 could also inhibited tumor cell proliferation and IC50 were 18.87?M and 63.77?M.We also verified the function area of B9 was SH2 domain in STAT3 using Biacore assay.Conclusion: After structure-based drug design,we have got(1)A small molecule 81 that inhibited CDK2 activity.After chemical modification of compound 81,we got ATP competitive inhibitor WZ-26 which could inhibit the proliferation of liver cancer cells;(2)A small molecule B9 that inhibited STAT3 activity.It could inhibit the proliferation of breast cancer and prostate cancer cells.The function area of B9 was SH2 domain. |
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