Biomimetic trapping cocktail to screen reactive metabolites: use of an amino acid and DNA motif mixture as light/heavy isotope pairs differing in mass shift
Biomimetic trapping cocktail to screen reactive metabolites: use of an amino acid and DNA motif mixture as light/heavy isotope pairs differing in mass shift
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用于筛选反应性代谢物的仿生捕获混合物：使用氨基酸和 DNA 基序混合物作为质量位移不同的轻/重同位素对
DOI：
10.1007/s00216-018-1057-z
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发表时间：
2018
影响因子：
4.3
通讯作者：
Tomoyuki Oe
中科院分区：
文献类型：
--
作者：
Shuto Hosaka;Takuto Honda;Seon Hwa Lee;Tomoyuki Oe
可以通过代谢转化为活性亲电产物的候选药物，如环氧化物、苯二酚和亚硝基化合物，尤其受到关注，因为随后与生物大分子的共价结合可能导致不良药物反应，如过敏反应、肝毒性和遗传毒性。已经报道了几种筛选反应性代谢物的策略，例如与放射性同位素标记药物的共价结合分析和随后的LC-MS/MS分析的捕集方法。其中，使用谷胱甘肽的捕集方法是最常见的，特别是在药物开发的早期阶段。然而，谷胱甘肽的半胱氨酸并不是体内唯一的亲核部位；赖氨酸、组氨酸、精氨酸和DNA碱基也是亲核的。事实上，谷胱甘肽捕捉法往往忽略了几种类型的反应性代谢物，如醛、酰基葡萄糖醛酸类和亚硝基化合物。在这里，我们介绍了一种筛选活性代谢物的替代方法：将简化氨基酸基序的轻同位素和重同位素与DNA基序的混合物用作仿生捕获鸡尾酒。该混合物由[2H0]/[2H3]-1-甲基胍(精氨酸基序，Δ3×Da)、[2H0]/[2H4]-2-巯基乙醇(半胱氨酸基序，Δ4×Da)、[2H0]/[2H5]-4-甲基咪唑(组氨酸基序，Δ5×Da)、[2H0]/[2H9]-正丁胺(赖氨酸基序，Δ9×Da)和[13C0，15N0]/[13C1，15N2]-2‘-脱氧鸟苷(Δ，DNA3×Da)组成。使用质量标签触发的数据相关采集来寻找特征双对峰，然后使用MS/MS对光同位素峰进行特异性鉴定。为了验证该策略，使用体外微生物学实验对42种模型药物进行了检验。图形摘要仿生捕获鸡尾酒筛选活性代谢物
Candidate drugs that can be metabolically transformed into reactive electrophilic products, such as epoxides, quinones, and nitroso compounds, are of special concern because subsequent covalent binding to bio-macromolecules can cause adverse drug reactions, such as allergic reactions, hepatotoxicity, and genotoxicity. Several strategies have been reported for screening reactive metabolites, such as a covalent binding assay with radioisotope-labeled drugs and a trapping method followed by LC–MS/MS analyses. Of these, a trapping method using glutathione is the most common, especially at the early stage of drug development. However, the cysteine of glutathione is not the only nucleophilic site in vivo; lysine, histidine, arginine, and DNA bases are also nucleophilic. Indeed, the glutathione trapping method tends to overlook several types of reactive metabolites, such as aldehydes, acylglucuronides, and nitroso compounds. Here, we introduce an alternate way for screening reactive metabolites as follows: A mixture of the light and heavy isotopes of simplified amino acid motifs and a DNA motif is used as a biomimetic trapping cocktail. This mixture consists of [2H0]/[2H3]-1-methylguanidine (arginine motif, Δ 3 Da), [2H0]/[2H4]-2-mercaptoethanol (cysteine motif, Δ 4 Da), [2H0]/[2H5]-4-methylimidazole (histidine motif, Δ 5 Da), [2H0]/[2H9]-n-butylamine (lysine motif, Δ 9 Da), and [13C0,15N0]/[13C1,15N2]-2′-deoxyguanosine (DNA motif, Δ 3 Da). Mass tag triggered data-dependent acquisition is used to find the characteristic doublet peaks, followed by specific identification of the light isotope peak using MS/MS. Forty-two model drugs were examined using an in vitro microsome experiment to validate the strategy.Graphical abstractBiomimetic trapping cocktail to screen reactive metabolites