Laine Jaana E, Häkkinen Merja R, Auriola Seppo, Juvonen Risto O, Pasanen Markku
University of Eastern Finland, Faculty of Health Sciences, School of Pharmacy, Yliopistonranta 1, P.O. Box 1627, FIN-70211 Kuopio, Finland.
Toxicol Rep. 2015 Jul 9;2:1024-1032. doi: 10.1016/j.toxrep.2015.07.002. eCollection 2015.
Qualitative trapping profile of reactive metabolites arising from six structurally different compounds was tested with three different d-peptide isomers (Peptide 1, gly-tyr-pro-cys-pro-his-pro; Peptide 2, gly-tyr-pro-ala-pro-his-pro; Peptide 3, gly-tyr-arg-pro-cys-pro-his-lys-pro) and glutathione (GSH) using mouse and human liver microsomes as the biocatalyst. The test compounds were classified either as clinically "safe" (amlodipine, caffeine, ibuprofen), or clinically as "risky" (clozapine, nimesulide, ticlopidine; i.e., associated with severe clinical toxicity outcomes). Our working hypothesis was as follows: could the use of short different amino acid sequence containing d-peptides in adduct detection confer any add-on value to that obtained with GSH? All "risky" agents' resulted in the formation of several GSH adducts in the incubation mixture and with at least one peptide adduct with both microsomal preparations. Amlodipine did not form any adducts with any of the trapping agents. No GSH and peptide 2 and 3 adducts were found with caffeine, but with peptide 1 one adduct with human liver microsomes was detected. Ibuprofen produced one Peptide 1-adduct with human and mouse liver microsomes but not with GSH. In conclusion, GSH still remains the gold trapping standard for reactive metabolites. However, targeted d-peptides could provide additional information about protein binding potential of electrophilic agents, but their clinical significance needs to be clarified using a wider spectrum of chemicals together with other safety estimates.
使用小鼠和人肝微粒体作为生物催化剂，用三种不同的d -肽异构体（肽1，甘氨酸 - 酪氨酸 - 脯氨酸 - 半胱氨酸 - 脯氨酸 - 组氨酸 - 脯氨酸；肽2，甘氨酸 - 酪氨酸 - 脯氨酸 - 丙氨酸 - 脯氨酸 - 组氨酸 - 脯氨酸；肽3，甘氨酸 - 酪氨酸 - 精氨酸 - 脯氨酸 - 半胱氨酸 - 脯氨酸 - 组氨酸 - 赖氨酸 - 脯氨酸）和谷胱甘肽（GSH）测试了六种结构不同化合物产生的反应性代谢物的定性捕获谱。测试化合物被分类为临床“安全”（氨氯地平、咖啡因、布洛芬）或临床“有风险”（氯氮平、尼美舒利、噻氯匹定；即与严重临床毒性结果相关）。我们的工作假设如下：在加合物检测中使用含有不同短氨基酸序列的d -肽能否为用GSH获得的结果带来任何附加价值？所有“有风险”的药物在孵育混合物中都导致形成了几种GSH加合物，并且与两种微粒体制剂至少形成一种肽加合物。氨氯地平与任何捕获剂都未形成任何加合物。咖啡因未发现GSH以及肽2和3的加合物，但在人肝微粒体中检测到与肽1形成一种加合物。布洛芬在人和小鼠肝微粒体中产生一种肽1 -加合物，但与GSH未产生。总之，GSH仍然是反应性代谢物的金标准捕获剂。然而，靶向d -肽可以提供有关亲电试剂蛋白质结合潜力的额外信息，但其临床意义需要使用更广泛的化学物质以及其他安全性评估来阐明。
