Study On Design,Synthesis And Anti-tumor Avtivity Of Pyrazolpyrimidines As CDK2 Inhibitor

The cell cyclin dependent kinase(CDKs) plays an important role in regulating cell division cycle. It can dominate the start, conduct and termination of cell cycle. Through occupying the active cavity of ATP binding pocket, CDKs inhibitors can prohibit the activation of protein kinase complexes, induce apoptosis and thus acheive the goal of anti-tumor. As a new breakthrough for targeted cancer therapy, the development of new inhibitors of CDKs is currently one of the goals pursued by scientists. Some of the CDK2 inhibitors were investigated clinically for their potential as anti-cancer agents. They show better anti-tumor effects and possess a good prospects in the treatment of cancer. Based on the study of structure-activity relationships of the CDK2 inhibitors, a serious of pyrazolpyrimidine derivatives have been designed and synthesis as CDK2 inhibitors. The studies on the inhibitory activities of title compounds will be theoretical and experimental basis for the development of tumor targeting inhibitors.The structural design and dock of CDK2 inhibitor with pyrazolpyrimidin e were conducted. A series of new CDK2 inhibitors with pyrazolpyrimidine skeleton were designed by analyzing the binding mode of the ATP and the known inhibitors with CDK2. The inhibitory activities of 32 designed compounds were virtually screen by docking algorithm on the platform of Autodock 4.0. According to the binding free energy(ΔG) and the inhibit constant(Ki), the influence of different substitutes on the docking results was investigated. And the stuctures of 15 pyrazolpyrimidine were confirmed.The synthesis of the pyrazolpyrimidines as CDK2 inhibitors was carried out. Based on the review of synthesis methods, the synthesis route of preparing the pyrazolpyrimidines was determined. Using ethoxymethylene malononitrile and tert-butyl hydrazine as starting materials, the target compounds were obtained via a series of reaction, including the Paal-Knorr cyclization, pyrimidine ring cyclization, halo-substituted reaction, Suzuki coupling reaction and aldol condensation. The influence factors, such as solvent, tempterure, catalyst and condensing agent on the reaction, were invented in details. The synthesizing condition of steps was optimized. Fifteen pyrazolpyrimidines were synthesized by adopting microwave irradiation with the yields of 30%. All structures of these new compounds were confirmed by 1H NMR, 13 C NMR and HRMS spectrum.The anti-tumor activity of pyrazolpyrimidines as CDK2 inhibitor was evaluated. CCK-8 was used to detected the inhibitivty activity of CDK2 by pyrazolpyrimidine derivatives in the breast cancer cell line MCF-7. The results indicated that the designed compounds possessed acceptable inhibitory activity with IC50 about 10 μM.