摘要
研究补骨脂定在人肝微粒体(HLM)和肠微粒体(HIM)的代谢活性,明确参与补骨脂定代谢的细胞色素P450酶(CYPs)和尿苷二磷酸葡萄糖醛酸转移酶(UGTs)及补骨脂定体外代谢的种属差异。将不同浓度的补骨脂定溶液分别与HLM和HIM共同孵育,经HLM孵育可以产生2个氧化产物(M1和M2)和2个葡萄糖醛酸结合物(G1和G2),在HIM中仅产生M1和G1;在HLM和HIM中,补骨脂定代谢生成M1的固有清除率(CL_(int))分别为104.3、57.6μL·min^(-1)·mg^(-1),生成G1的CL_(int)分别为543.3、75.9μL·min^(-1)·mg^(-1)。利用12种CYPs和12种UGTs酶,分别与不同浓度的补骨脂定溶液共同孵育,结果显示,CYP1A1(39.5μL·min^(-1)·mg^(-1))、CYP2C8(88.0μL·min^(-1)·mg^(-1))、CYP2C19(166.7μL·min^(-1)·mg^(-1))、CYP2D6(9.1μL·min^(-1)·mg^(-1))是生成M1的关键CYPs代谢酶,而CYP2C19(42.0μL·min^(-1)·mg^(-1))是生成M2的重要亚型酶;UGT1A1(1184.4μL·min^(-1)·mg^(-1))、UGT1A7(922.8μL·min^(-1)·mg^(-1))、UGT1A8(133.0μL·min^(-1)·mg^(-1))、UGT1A9(348.6μL·min^(-1)·mg^(-1))、UGT2B7(118.7μL·min^(-1)·mg^(-1))重点参与G1的生成,而UGT1A9(111.3μL·min^(-1)·mg^(-1))是G2生成的关键亚型酶。采用猴肝微粒体(MkLM)、大鼠肝微粒体(RLM)、小鼠肝微粒体(MLM)、狗肝微粒体(DLM)和猪肝微粒体(MpLM),分别与不同浓度的补骨脂定溶液共同孵育,结果显示,补骨脂定的Ⅰ相代谢和葡萄糖醛酸化代谢均表现出显著的种属差异。总体来说,补骨脂定在肝肠微粒体均可以发生较强的代谢;CYP1A1、CYP2C8、CYP2C19、CYP2D6与UGT1A1、UGT1A7、UGT1A8、UGT1A9、UGT2B7是参与其代谢的关键亚型酶;大鼠和猪分别是研究补骨脂定Ⅰ相代谢和葡萄糖醛酸化代谢合适的模式动物。
This study aims to investigate metabolic activities of psoralidin in human liver microsomes(HLM)and intestinal microsomes(HIM),and to identify cytochrome P450 enzymes(CYPs)and UDP-glucuronosyl transferases(UGTs)involved in psoralidin metabolism as well as species differences in the in vitro metabolism of psoralen.First,after incubation serial of psoralidin solutions with nicotinamide adenine dinucleotide phosphate(NADPH)or uridine 5’-diphosphate-glucuronic acid(UDPGA)-supplemented HLM or HIM,two oxidic products(M1 and M2)and two conjugated glucuronides(G1 and G2)were produced in HLM-mediated incubation system,while only M1 and G1 were detected in HIM-supplemented system.The CL_(int) for M1 in HLM and HIM were 104.3,and 57.6μL·min^(-1)·mg^(-1),respectively,while those for G1 were 543.3,and 75.9μL·min^(-1)·mg^(-1),respectively.Furthermore,reaction phenotyping was performed to identify the main contributors to psoralidin metabolism after incubation of psoralidin with NADPH-supplemented twelve CYP isozymes(or UDPGA-supplemented twelve UGT enzymes),respectively.The results showed that CYP1 A1(39.5μL·min^(-1)·mg^(-1)),CYP2 C8(88.0μL·min^(-1)·mg^(-1)),CYP2 C19(166.7μL·min^(-1)·mg^(-1)),and CYP2 D6(9.1μL·min^(-1)·mg^(-1))were identified as the main CYP isoforms for M1,whereas CYP2 C19(42.0μL·min^(-1)·mg^(-1))participated more in producing M2.In addition,UGT1 A1(1184.4μL·min^(-1)·mg^(-1)),UGT1 A7(922.8μL·min^(-1)·mg^(-1)),UGT1 A8(133.0μL·min^(-1)·mg^(-1)),UGT1 A9(348.6μL·min^(-1)·mg^(-1))and UGT2 B7(118.7μL·min^(-1)·mg^(-1))played important roles in the generation of G1,while UGT1 A9(111.3μL·min^(-1)·mg^(-1))was regarded as the key UGT isozyme for G2.Moreover,different concentrations of psoralidin were incubated with monkey liver microsomes(MkLM),rat liver microsomes(RLM),mice liver microsomes(MLM),dog liver microsomes(DLM)and mini-pig liver microsomes(MpLM),respectively.The obtained CL_(int) were used to evaluate the species differences.Phase Ⅰ metabolism and glucuronidation of
作者
秦子飞
张贝贝
邢晗
王培乐
杨晶
张晓坚
姚志红
姚新生
QIN Zi-fei;ZHANG Bei-bei;XING Han;WANG Pei-le;YANG Jing;ZHANG Xiao-jian;YAO Zhi-hong;YAO Xin-sheng(Department of Pharmacy,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Development Ministry of China,Jinan University,Guangzhou 510632,China;College of Pharmacy,Jinan University,Guangzhou 510632,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2021年第13期3410-3421,共12页
China Journal of Chinese Materia Medica
基金
国家自然科学基金青年基金项目(81903704,81703799)
广东省基础与应用基础研究基金项目(2019A1515011285)
新药研究国家重点实验室开放基金项目(SIMM1903KF-07)。
