Design,Synthesis And Biological Activity Study Of [1,2,4] Triazol [4,3-a] Pyridine Derivatives As Potential C-Met Inhibitors

Cancer is still one of the world's most common and most serious diseases which directly endanger human's lives.In the past half-century,a large number of resources had been invested to the research of cancer treatment and great progress of many aspects had been gained,given to which tumor signaling networks have been gradually explained and completed.It was shown that many human tumors were related with abnormal activation of tyrosine kinase route.Given to the existing data,it is suggested that more than 50%of the proto-oncogene has protein tyrosine kinase activity,in which c-Met is one of the most important aisle of protein tyrosine kinases,therefore the development of c-Met specific drugs are currently a hot topic of targeted cancer therapy research.To obtain high efficiency,low toxicity and easy for industrial synthesis of specific small molecule inhibitors of c-Met,we used published data of triazole c-Met inhibitor to design for triazole c-Met inhibitor:firstly,optimize the triazole nucleus,the triazole pyridazine and triazole pyrazine nucleus were optimized as triazolopyridine nucleus,and the fluorine atom substituent was introduced to an 8-position of nucleus,and further investigation was made on the effect of position substituent on the compound activity.Secondly,maintain the nucleus 6-position as 1'-methyl-pyrazol-5-yl,while using a sulfur atom as a connecting bridge,to obtain high activity in c-Met inhibitor while simplifying the synthesis process.Finally,main investigation was made to the 3-position and 4-position substituent group of quinoline ring;halo,alkyl,substituted phenyl group,amide group,ether group,morpholine,quinoline and thiazole group were tried to introduce into the 3-position and 4-position.Through experiments,16 compounds were observed with c-Met inhibitor ICso<10 nM,in which 11 compounds IC50<5 nM;while 16 compounds were observed with gastric cancer cell SNU-5 inhibition IC50<10 nM,in which 13 compounds IC50<5 nM and 7 compounds IC50<1 nM.Based on the work above,we made further optimization design for substituent group of[1,2,4]triazolo[4,3-a]pyridine derivatives;the quinoline ring 3-and 4-position open chain substituent group were replaced with cyclic substituent,such as dioxins,dioxepino,pyrrolo morpholino,morphinone,morpholino,thiazoles and oxazolone substituents plurality heterocyclic group etc.Compound 49 was gained with the highest bioactive with IC50 for c-Met inhibition activity is 0.7 nM,and it's IC50 for gastric cancer cell SNU-5 inhibition is 2 nM.Further experimental study of in vitro enzymatic activity for compound 49 showed that:The inhibition activity of compound 49 on c-Met kinase was highly selective and specific.In vitro cell viability experiment,compound 49 significantly inhibited the proliferation of c-Met high expression cancer cell line caused by the gene amplification of c-Met,while it had no activity on c-Met-negative cell line and c-Met-positive cell line which do not produce HGF.In the experiments for proliferation inhibition and selectivity of measured cell line,the effect of compound 49 showed an equivalent effect to the positive control SGX-523 and JNJ-38877605.In some cancer cells,compound 49 showed a higher activity.In vivo pharmacokinetic study of rats,compared to the positive control JNJ-38877605,compound 49 showed higher plasma concentration,longer half-life,longer residence time in vivo,lower clearance rate suggested that compound 49 had a good pharmacokinetic properties performance.The results of vivo efficacy study clearly indicated that compound 49 had a better inhibiting activity than SGX-523 in a dose-dependent manner.Long-term toxicity studies were estimated that the No Observed Adverse Effect Level(NOAEL)to be 225(mg/kg)/day and the NOAEL in...