Design,Synthesis And Biological Evaluation Of Pyrazolopyrimidines As Akt Inhibitors

Malignant tumors seriously endanger human life and health and damage human life and property.Although my country is committed to the research of malignant tumors and has achieved certain results,the discovery of new target molecules and the development of new therapeutic technologies are still crucial.The family of protein kinases with more than 500 members represents the primary therapeutic target for drug development.To date,more than 150 protein kinases have been shown to be involved in the development of human diseases,particularly cancer,various immunodeficiency,endocrine disorders and neurodegenerative diseases.So it is very important to develop potent kinase inhibitors.So far,most of the studies are small molecule inhibitors.The sustained activationof PI3K/Akt signaling pathway is clo sely related to the growth and survival of cancer cells.Blocking the sustained activation of this signaling pathway provides a new strategy for targeted treatment of cancer,and the inhibitors acting on this signaling pathway become potential drugs for cancer treatment.Akt is a threonine kinase activated by the recruitment of phosphatidylinositol into plasma membrane,which plays a key role in cell apoptosis and survival,cell proliferation,vascular proliferation and regulation of cell cycle and metabolism,as well as resistance to chemotherapy and radiotherapy.It has been found that the Akt gene locus is similar to the immunoglobulin heavy chain gene locus,and the frequent heterotopia and inversion of the latter locus are closely related to the occurrence of leukemia and lymphoma,suggesting that the activation and regulation of Akt are related to the occurrence of tumor.Therefore,Akt can be regarded as an important target of cancer treatment and even cancer prevention.GDC0068 is a selective,competitive Akt inhibitor with IC50 of 5 nM,18 nM,and 8 nM for inhibiting Aktl,Akt2,and Akt3,respectively,which is 620 times more selective than PKA.Many GDC0068 derivatives have also been obtained in the laboratory's previous work.Among them,compound 14g showed well growth inhibitory activity on Akt kinase and six lymphoma cells.In the design of the target compounds,first,replace the parent core of compound 14g with a pyrazolopyrimidine structure to examine the effect of different parent core structures on compound activity;Secondly,replace the N-methylpiperidine at the alpha position of the carbonyl group with N-substituted chains of different lengths to explore the effect of its combination with Acid Hole;finally,there are different substituents attached to the benzene ring to examine the different substituent effect on compound activity.Twenty-four of novel pyrazolopyrimidine compounds were designed and synthesized and confirmed by 1H-NMR,13C-NMR and High Resolution Mass Spectrometry(HRMS).By measuring the inhibitory activity of compounds on Akt kinase,it was found that some compounds showed better inhibitory activity on Aktl kinase.Compounds GK3,GK4 and GK16 had stronger inhibitory activity on Aktl kinase,with IC50 values of 23.54,20.55 and 15.03 nM,respectively.Better than the positive control GDC0068(IC50=33.62 nM).In the test results of the growth inhibitory activity of MCL cells,we found that most of the compounds had good growth inhibitory activity on six lymphoma cells,and compounds GK4 and GK16 had better inhibitory activity on six lymphomas cells than that of positive control Ibrutinib(IBN).And the inhibitory activity of GK3 and GK15 in four cell lines(Rec-1?Maver-1.Z-138 and OCI-LY10)was better than that of the positive control IBN.GK16,which has a strong inhibitory effect on Akt,also showed a slightly stronger antiproliferative activity of six lymphomas cells than IBN 2-10 times.It is possible to further study the selectivity of GK16 for Akt kinase,biological activity evaluation and mechanism of various tumor cells.