Design,Synthesis And Antitumor Activity Study Of Novel Heterocyclic Pyrimidine Derivatives As Small Molecule C-Met Inhibitors

HGF/c-Met signaling pathway is an important branch of tyrosine kinase signaling pathways.Studies have revealed that HGF is abnormally high expression in various tumor tissues and closely related to tumor invasion and metastasis.Compared with the high recurrence of traditional treatment methods,molecular targeted therapy contain many advantages such as reducing the dosage,improving the therapeutic effect,reducing toxic and side effects.Therefore,the small molecules of c-Met targeting inhibitor's development has become an important strategy in the field of cancer treatment.In this dissertation,based on the structure-activity relationship of class II small molecule c-Met inhibitors,the characteristics of 5-atom fragments were preserved and modified with triazole and pyridazinone as active fragments.For the nucleus part,quinolone structure was substituted with pyrrolo-pyrimidines,pyrazolopyrimidines,and thienopyrimidines by bioisosterism and skeleton transitions.As a results,97 novel compounds were finally reported.The structures of all the target compounds were confirmed by ¹H NMR and MS spectra,and the structures of some compounds were confirmed by ¹³C NMR.For the pharmacological activity part,A549,MCF-7,and HepG2 were used as test cell lines,and Foretinib was used as a positive control drug.Cytotoxic tests were performed on the 97 target compounds.The results showed that these compounds had good selectivity for HepG2 cell lines,24 of them showed better inhibitory activity against one or more tumor cell lines,which was superior or comparable to Foretinib.Among them,three selected compounds WLX-9?c-Met:790 nM?,WLX-58?c-Met:16 nM?and WLX-91?c-Met:19 nM?were valued against c-Met,Flt-3,and VEGFR-2,c-Kit and EGFR kinase by enzyme inhibitory activity tests.The results showed that these compounds exhibited moderate to superior inhibitory activity on c-Met kinases and exhibiting superior c-Met kinase selectivity.In addition,the data of the optimal compounds WLX-58 and WLX-91 showed that they could induce early apoptosis of HepG2 cells in a concentration-dependent manner by acridine orange?AO?staining and Annexin V-FITC/PI flow cytometry apoptosis assay.These results indicate that the designed compounds are 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.The"5-atom"moiety plays an important role in the activity of the compound;the fluorine atom introduced into the aminophenoxy group keeps the activity of the compound unchanged or even enhanced;the compounds?WLX-36^WLX-97?contained a thienopyrimidine structure showed more activity than that of pyrrole/pyrazolopyrimidine structure compounds?WLX-1^WLX-35?;What's more,the activity of thiophene nucleus contained 6-oxo-pyridazinone side chain?WLX-64^WLX-92?was similar to that of compounds containing triazole side chain?WLX-36^WLX-63?;in addition,among these pyridazinone compound,6-oxo-pyridazinone structural compound?WLX-64^WLX-92?showed more activity than that of 4-oxo-pyridazinone structural compound?WLX-93^WLX-97?;in addition,the position of the substituent on the hydrophobic segment and the kind of the substituent have a great influence on the activity.In this dissertation,the structure-activity relationship and the mechanism of action of heterocyclic pyrimidines were primarily analyzed and discussed,which pointed out the direction for future in-depth study of these inhibitors.