Design,Synthesis And Biological Activity Research Of Antitumor Compounds Based On C-Met Targets Inhibitors

The HGF/c-Met signaling pathway is highly expressed in a variety of tumor tissues and plays an important role in tumorigenesis,development,invasion and metastasis.Therefore,the search for novel and efficient small molecule inhibitors targeting HGF/c-Met signaling pathway has become a research hotspot.This paper mainly introduces the research situation of HGF/c-met inhibitors,on which basis the design,synthesis and anti-tumor activity in vitro of new small-molecule c-met inhibitors are studied.According to their chemical structures and the binding mode with c-Met protein,small molecule c-Met inhibitors are divided into two types: Class I and Class II.By summarizing the structures of Class I c-Met inhibitors,we found that the active medicinal components can be divided into three main structures such as triazolopyrazine,triazolopyrazine and triazolopyrimidine.According to the published structure of Class II c-Met inhibitors,the structure-activity relationship was initially summarized.The positively active Class II c-Met inhibitor Foretinib was selected as the positive control drug.On this basis,the quinoline nucleus was retained,and the "5 atom" fragment was modified by the active structure pyridazinone.The quinoline derivative L-1~L-32 containing the pyridazine structure was designed.In addition,active pharmacophores of Class I and Class II c-Met inhibitor were used to develop more active small molecule c-Met inhibitors.Therefore,the triazolopyrazine structure in the Class I c-Met inhibitor was introduced into the type II inhibitor to replace the quinoline nucleus and retain the activity of pyrazoline structure.Thus,L-33~L-52 derivatives of trizolpyrazoline containing pyrazine structures were designed.A novel five-membered heterocyclic ring was introduced into the "5 atom" moiety,and the benzene ring extending into the hydrophobic cavity was substituted with a pyridine or thiophene group,and designing the containing the five-membered heterocycle structure triazolopyrazine derivatives.Thus,triazole and pyrazine derivatives L-53~L-77 containing five heterocyclic rings were designed.According to the principle of bioisosterism and skeleton transitions,the triazolopyrazine structure was replaced by triazolopyridazine and triazolopyrimidine structure to design the triazolopyridazine/pyrimidine derivatives L-78~L-97 containing the five-membered heterocycle structure.All of the 97 synthesized target compounds were tested for their cytotoxic activities against A549,MCF-7,Hep G2 and Hela cell lines by the MTT method,using Foretinib as positive controls.Most of the target compounds showed moderate to excellent anti-cell proliferation activity against one or more tested tumor cells.To study the effects of compound targets,homogeneous phase-resolved fluorescence(HTRF)was used for the determination of c-Met kinase activity in the target compound using Foretinib as a positive control,17 of which showed better or similar to Foretinib Met kinase activity.In addition,concentration-dependent,acridine orange(AO)staining,apoptosis and cycle tests were performed on the preferred compounds L-32,L-51,L-76 and L-79.The data indicated that they can induce apoptosis in tumor cells in a concentration-dependent manner,indicating that the designed compounds were a class of potent c-Met inhibitors.According to the results of anti-tumor activity of compounds in vitro,their structure-activity relationship was initially summarized.(1)Substitution of the quinoline nucleus with triazolopyrazine,triazolopyrazine,and triazolopyrimidine structure has little effect on the activity of the compound.(2)The fluorine atom introduced into the aminophenoxy group keeps the activity of the compound unchanged or even enhanced.(3)The difference in the substituents on the side chain benzene ring has a great influence on the activity of the compound.Generally,the stronger the electron withdrawing ability of the substituents,the better the activity of the compound;when the benzene ring has a double electron withdrawing group,the activity is better than the single electron withdrawing group;The compound has the best activity when there is no substituent on the benzene ring.(4)Replace the benzene ring on the side chain with pyridine or thiophene structure.Since 2-pyridine can form bidentate hydrogen bond with the amino acid residue together with the "5 atom" part,its activity is better than that of benzene ring or other structures;substitution of the phenyl ring with a thiophene structure has no significant effect on the antitumor activity in vitro of the target compound.Combined with the docking results of the target compounds and c-met kinase,this paper made a preliminary analysis and discussion on the structure-activity relationship and the mechanism of action of the target compounds,which pointed out the direction for further in-depth study of these inhibitors.