Design,Synthesis,Biological Evaluation And Molecular Simulation Of Novel Src Family Kinase Inhibitors

Glioblastoma(GBM)is the most common and malignant primary brain tumor in the central nervous system.Despite the use of high-intensity treatment such as surgery,radiotherapy,chemotherapy,patient prognosis is still very poor,only 4%-5%of the five-year survival rate,which urgently needs more effective treatment.Recent studies suggest that the Src family kinases(SFKs)may be promising molecular targets for GBM therapy.SFKs participate in the regulation of cell proliferation,survival,invasion,angiogenesis and other physiological processes,high expression and excessive activation in GBM,induce a variety of tuma effects in GBM,including reduced apoptosis,promoted vascular production and cell proliferation.Inhibition of SFKs can reduce GBM growth,vitality,and migration.Therefore,we believe that the discovery and study of SFK inhibitors have important theoretical value and potential application prospects for individualized treatment of GBM.Based on the cocrystal complexes of Src family kinases with small molecule PP2 and approved drug Dasatinib,two series of compounds were designed with imidazolone[4,5-c]pyridine and 3-aminopyrazole as the parent nucleus,respectively.The inhibitory activity of compounds against Src and Fyn kinase of SFKs member and the anti-proliferation activity against tumor cells with high expression of SFKs were evaluated.The physicochemical characteristics of the compounds and their binding characteristics with SFKs were analyzed by molecular simulation.According to the structural characteristics of SFK inhibitor PP2,we first designed and synthesized twelve N1 diffrent substituted imidazolone[4,5-c]pyridine compounds,which did not show obvious inhibitory effect on Src and Fyn kinase.Cell antiproliferation experiments show that compounds 2a-c,2e-h significantly inhibited the proliferation of U87 GBM cell line,suggesting that tumor inhibition may be achieved by inhibiting other targets.Then on the basis of the synthesized compounds,we continued to carry out structural modification to synthesize ten N1 and N3 substituted imidazolone[4,5-c]pyridine compounds.Compound 1d,le,1f showed obvious inhibitory effect on Src and Fyn kinase.Further optimization on the structure of 1d to synthesize nine compounds with different modifications of the N3 on the imidazolone ring.Compound 1k,1q,1s exhibit significant Src and Fyn kinase inhibitory effects.Four compounds with the best kinase activity(1d,1e,1q,1s)were selected to test their antiproliferative activity against six tumor cell lines characterized by SFKs hyperactivation.1s had the best antiproliferative activity(comparable to PP2).Molecular simulation results show that compound 1s meet the physical and chemical properties of central nervous system drugs,and can stably bind to the ATP binding sites of SFKs with the expected conformation.Compound 1s as a novel SFK inhibitor antiGBM animal in vivo activity is worth further study further.According to the structural characteristics of SFK inhibitor Dasatinib,we designed and synthesized twelve 3-aminopyrazole compounds.Unfortunately,these compounds did not show significant inhibitory activity against Src and Fyn kinases at 1 μM concentration,suggesting that further structural modification and optimization are needed.In summary,forty-three small molecule compounds were designed and synthesized by using the rational drug design strategy,including thirty-one imidazolone[4,5-c]pyridine compounds and twelve 3-aminopyrazole compounds.The in vitro biological activity evaluation and molecular simulation were carried out,Our study provided candidate compounds and experimental basis for finding new SFK inhibitors.