新靶点SGLT2选择性抑制剂是目前2型糖尿病小分子口服治疗药物的开发热点。Dapagliflozin是2014年刚被美国FDA批准上市的第二个SGLT2抑制剂，由于担心其潜在的致癌风险，FDA要求对接受治疗的患者定期监测膀胱癌和肝脏毒性等风险指标。鉴于药物毒性与其代谢毒物的相关性，本项目科学地提出Dapagliflozin类SGLT2抑制剂在体内代谢过程中可能会产生临床副作用的致毒物。因此，为提高该类SGLT2抑制剂安全性，探索Dapagliflozin类药物产生临床毒性的原因，阐明毒代产物（次甲基醌类）可能的结构和药理毒理；并利用化学合成和生物酶CYP450催化氧化两种手段，揭示其代谢警戒物4和5（苯酚包括苯氧烷基）与其产生可能的致毒物（次甲基醌类）的机制；并在前期已发表在J Med Chem工作基础之上，创新性氟代结构优化，通过阻断活性代谢位点，研发高效安全的新型SGLT2抑制剂。

Selective SGLT2 inhibitor as new target is a hot point of oral small molecule therapy for type 2 diabetes. Dapagliflozin has just been approved by the U.S. FDA as the second SGLT2 inhibitor in 2014. Because of its potential risk of cancer, FDA requires patients undergoing treatment to regularly monitor the risk indicators of bladder cancer and liver toxicity. Considering the correlation between the toxicity and its toxic metabolites of drug, this proposal suggested that SGLT2 inhibitor maybe produce toxic metabolites in vivo metabolic processes, which caused clinical side effects. Therefore, the project aimed at improving the safety of such SGLT2 inhibitors, exploring the reason for the clinical side effects of Dapagliflozin, clarifying the structures and toxicology of toxic metabolites (methine quinone). The possible metabolism mechanism was revealed by chemical synthesis and CYP450 enzyme catalysis. On the basis of preliminary studies published in J Med Chem, new efficient and safe SGLT2 inhibitors were developed by innovative fluorinated structural optimization and blocking the active metabolic sites.

本项目对照原定预期成果和技术考核指标，重点围绕全球首个上市的新型抗2型糖尿病SGLT2抑制剂新药Dapagliflozin，聚焦其在临床用药中发现膀胱炎和肝脏损伤，甚至可能导致乳腺癌和膀胱癌病变毒副作用的风险增加这一科学问题，创新地制备并使用碳-13标记的Dapagliflozin来追踪代谢产物的质谱信息，以及利用谷胱甘肽捕捉潜在毒代物次甲基醌形成稳定结合物的性质，对Dapagliflozin及碳-13标记Dapagliflozin的体外肝微粒体酶氧化产物及SD大鼠体内胆汁代谢物分别使用高科技分析手段UHPLC-QTOF-MS/MS等进行检测分析，顺利得到了2种次甲基醌类毒代物结合谷胱甘肽的质谱信息，使预测得到了验证。此外，基于先导化合物Dapagliflozin分子结构存在3个潜在活性代谢位点，在前期氘代研究的基础上，创新地使用氟代的策略对其主要代谢位点进行优化设计，共合成了3个氟代目标化合物，并且完成了其体外活性和选择性与先导化合物Dapagliflozin的对比测试，为后续进行深入研究奠定了坚实的基础。