Design,Synthesis,and Biological Evaluation Of Pyrido[4,3-d]Pyrimidin-4(3H)-One/2,3-Dihydropyrimido[4,5-D]Pyrimidin-4(1H)-One Derivatives As Wee1 Inhibitors

Wee1 kinase is essential for controlling the cell cycle and repairing DNA damage.Inhibition of Wee1 abrogates cell cycle arrest,leading to replication stress,promoting mitotic catastrophe and subsequent cell death via the apoptotic program.Targeting Wee1 has been confirmed as an effective strategy for cancer therapy.However,clinical studies have found that Wee1 inhibitors have serious side effects,such as myelosuppression and gastrointestinal toxicity.Therefore,it is of great significance and value to develop novel Wee1 inhibitors with high efficiency and excellent selectivity.Enlightened by the compound A published in the literature and its binding mode with Wee1 protein,45 novel pyrido[4,3-d]pyrimidin-4(3H)-one derivatives were developed and synthesized using the strategy of scaffold hopping in this study.As a result,the majority of compounds exhibit potent inhibitory activities against Wee1.The compounds with strong Wee1 inhibitory activities also display moderate inhibitory activities onMV-4-11 cell line,and slightly weaker anti-proliferation activities on T47 D cell line.Mouse liver microsomal stability test demonstrates the good metabolic stability of compounds I-18,I-30,I-40,and I-41.The representative compound I-40 exhibits relatively high kinase selectivity against 9 kinase among the 11 selected kinase inhibition experiments.Mechanistic studies have shown that compound I-40 can inhibit the phosphorylation of downstream Y15 CDK1 site.Furthermore,a dose-dependent manner was observed in G0/G1 phase arrest and apoptosis of MV-4-11 cells,which has the potential for deeper investigation.Besides,Debio 0123 was also selected as the lead compound in this study.Based on the investigation of the substituents on the pyrimidinone N,a total of 112,3-dihydropyrimido[4,5-d]pyrimidin-4(1H)-one derivatives were designed and synthesized.However,these compounds show poor inhibitory activities against Wee1 in the preliminary experiment.Consequently,there is a requirement for more research into novel Wee1 inhibitors.