Nishimura Masuhiro, Naito Shinsaku
Division of Pharmacology, Drug Safety and Metabolism, Otsuka Pharmaceutical Factory Inc, Naruto, Tokushima, Japan.
Drug Metab Pharmacokinet. 2006 Oct;21(5):357-74. doi: 10.2133/dmpk.21.357.
Pairs of forward and reverse primers and TaqMan probes specific to each of 52 human phase I metabolizing enzymes (alcohol dehydrogenase, aldehyde dehydrogenase, aldehyde oxidase, dihydropyrimidine dehydrogenase, epoxide hydrolase, esterase, flavin-containing monooxygenase, monoamine oxidase, prostaglandin endoperoxide synthase, quinone oxidoreductase, and xanthene dehydrogenase) and 48 human phase II metabolizing enzymes (acetyltransferase, acyl-CoA:amino acid N-acyltransferase, UDP-glucuronosyltransferase, glutathione S-transferase, methyltransferase, and sulfotransferase) were prepared. The mRNA expression level of each target enzyme was analyzed in total RNA from single and pooled specimens of various human tissues (adrenal gland, bone marrow, brain, colon, heart, kidney, liver, lung, pancreas, peripheral leukocytes, placenta, prostate, salivary gland, skeletal muscle, small intestine, spinal cord, spleen, stomach, testis, thymus, thyroid gland, trachea, and uterus) by real-time reverse transcription PCR using an ABI PRISM 7700 Sequence Detection System. Further, individual differences in the mRNA expression of representative human phase I and II metabolizing enzymes in the liver were also evaluated. The mRNA expression profiles of the above phase I and phase II metabolizing enzymes in 23 different human tissues were used to identify the tissues exhibiting high transcriptional activity for these enzymes. These results are expected to be valuable in establishing drug metabolism-mediated screening systems for new chemical entities in new drug development and in research concerning the clinical diagnosis of disease.
制备了针对52种人类I相代谢酶（乙醇脱氢酶、醛脱氢酶、醛氧化酶、二氢嘧啶脱氢酶、环氧化物水解酶、酯酶、含黄素单加氧酶、单胺氧化酶、前列腺素内过氧化物合酶、醌氧化还原酶和黄嘌呤脱氢酶）和48种人类II相代谢酶（乙酰基转移酶、酰基辅酶A：氨基酸N-酰基转移酶、尿苷二磷酸葡萄糖醛酸基转移酶、谷胱甘肽S-转移酶、甲基转移酶和磺基转移酶）的正向和反向引物对以及TaqMan探针。使用ABI PRISM 7700序列检测系统，通过实时逆转录PCR分析了来自各种人类组织（肾上腺、骨髓、脑、结肠、心脏、肾脏、肝脏、肺、胰腺、外周血白细胞、胎盘、前列腺、唾液腺、骨骼肌、小肠、脊髓、脾脏、胃、睾丸、胸腺、甲状腺、气管和子宫）的单个和混合样本的总RNA中各目标酶的mRNA表达水平。此外，还评估了肝脏中代表性人类I相和II相代谢酶mRNA表达的个体差异。利用上述I相和II相代谢酶在23种不同人类组织中的mRNA表达谱来鉴定这些酶具有高转录活性的组织。这些结果有望在建立新药开发中新型化学实体的药物代谢介导的筛选系统以及疾病临床诊断研究中具有重要价值。
