Study On Design,Synthesis And Biological Activity Of Novel Tyrosine Kinase Inhibitors Based On C-met And ALK

c-Met and ALK protein all belong to receptor tyrosine kinases.Both of them are widely expressed in a variety of cells and participate in many kinds of physio logical activities.However,dysregulation of the HGF/c-Met or ALK signaling pathway by mutation,gene amplification and overexpression will ultimately result in the occurrence,development and metastasis of human cancers due to the disorder of cell growth and movement.It has been reported that HGF/c-Met and ALK involves in a series of human tumors,such as gastric cancer,lung cancer,liver cancer,ovarian cancer,breast cancer,neuroblastoma and anaplastic large cell lymphoma.Crizotinib is a novel dual small-molecular ATP-competitive inhibitor of ALK and c-Met,approved by the U.S.Food and Drug Administration(FDA)for the treatment of patients with ALK+ locally advanced or metastatic non-small cell lung cancer(NSCLC).So the structure of crizotinib is of great value for further research.Therefore,we designed and synthesized a series of novel compounds based on the structure of crizotinib and assessed their antiproliferative efficacy on human cancer cell lines.The two series of compounds were synthesized successively employing Williamson reaction with(R)-1-(2,6-dichloro-3-fluorophenyl)ethanol prepared via chemical resolution,Sonogashira reaction catalyzed by Pd(dppf)Cl2 and CuI,highly selective Huisgen-Click reaction,and finally removal of Boc with trifluoroacetic acid using 2-aminopyrazine and 3,6-dichloro pyridazine as starting materials respectively.In addition,their preliminary inhibition activities of tumor proliferation were evaluated against human neuroblastoma(SH-SY5Y)and human gastric carcinoma(SNU-5)cell line in vitro.Futhermore,molecular docking simulation studies were performed using Discovery Studio 4.0 CDOCKER Docking software in order to obtain a deep sight into the binding mechanism of synthesized compounds with target proteins ALK and c-Met.The results of antitumor activities showed that:(1)The inhibitory activities of pyrazine compounds against two tumor cell lines were better than that of the pyrazine series.(2)Pyrazine compound 2-10a showed best activity against SH-SY5Y cell line(IC50= 0.467 μmol·L-1)which was the same level as positive control Crizotinib(IC50 = 0.341 μmol·L-1);As for inhibitory activity against SNU-5 cel line,2-10e showed the best activity with IC50 value 148 nmol·L-1,a promising value despite not better than Crizotinib.(3)The antitumor activities of pyrazine compounds were not satisfactory,but they showed strong inhibitory activities against SNU-5 cell line.Among them,compound 3-9a showed the highest inhibitory activity against the two cell lines.Finally,molecular docking simulation studies were performed using Discovery Studio 4.0 CDOCKER Docking software.Docking results were then evaluated by the CDOCKER score function.The results showed that antitumor activities are basically in agreement with the Docking results evaluated by the CDOCKER score function.