Design And Synthesis Of Lactam Hypoxia Inducible Factor-1inhibitors Based On Metabolic Stability

Malignant cells have vigorous growth and metabolism,high oxygen consumption,abnormal growth of blood vessels in the tumor,abnormal blood supply and hypoxia microenvironment,which is one of the basic characteristics of solid tumor tissue.Hypoxia microenvironment continuously activates HIF-1 signaling pathway in malignant tumor cells and promotes the transcription and expression of a variety of downstream genes,including vascular epidermal growth factor（VEGF）,matrix metalloproteinases（MMPs）,glucose transporter（GLUT1）,epithelial mesenchymal transition（EMT）related transcription factors snail,twist,etc.,through promoting angiogenesis,tumor metabolic reprogramming,EMT,immune escape,inhibition of apoptosis and other pathways,It enhances the ability of tumor invasion and metastasis and resistance to radiotherapy and chemotherapy,and promotes the occurrence,development and metastasis of tumor.Therefore,the development of inhibitors targeting key signaling molecules in HIF-1 pathway has become one of the strategies in the field of anti-tumor metastasis drug research and development,and has attracted more and more attention.In the early stage,through fragment based drug design,the research group found that a class of arylformamide compounds had inhibitory effect on HIF-1 signal pathway,and the IC₅₀of AMSP-30m,the compound with the best activity,reached0.32μM。Preliminary pharmacological evaluation showed that compound AMSP-30m could inhibit tumor cell invasion,migration and angiogenesis,and inhibit lung metastasis of breast cancer cells in nude mice.However,in the follow-up evaluation of drug properties,we found that AMSP-30m had poor stability on liver microsomal metabolism.Based on improving the metabolic stability of AMSP-30m,the amide bond in its structure was changed to lactam to improve the metabolic stability.In this paper,four series of lactam rings were designed and synthesized,including benzodihydroxazinone（Series A）,dihydroisoquinolone（Series B）,benzozazanone（Series C）,and benzodihydroxyzazanone（Series D）.A total of 62 compounds were synthesized.After obtaining the target compound,we screened the target compound for inhibiting the activity of HIF-1 promoter by double reporter gene method,and found that 8 compounds were in 10μThe inhibition rate was more than 80%at M concentration.Furthermore,we tested the metabolic stability of liver microsomes of these compounds with good activity.The metabolic stability of liver microsomes was greatly improved by most compounds,and the half-life of five compounds was more than 100 min.In conclusion,a series of benzolactam compounds were designed and synthesized with arylformamide HIF-1 inhibitors as the guide and the idea of ring merging design.Compounds with HIF-1 inhibitory activity equivalent to that of the lead and greatly improved metabolic stability were obtained to provide subsequent evaluation of antitumor transfer pharmacological activity.