Characterisation of flucloxacillin and 5-hydroxymethyl flucloxacillin haptenated HSA in vitro and in vivo
Characterisation of flucloxacillin and 5-hydroxymethyl flucloxacillin haptenated HSA in vitro and in vivo
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DOI：
10.1002/prca.200800222
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发表时间：
2009-06-01
影响因子：
2
通讯作者：
Park, B. Kevin
中科院分区：
文献类型：
--
作者：
Jenkins, Rosalind E.;Meng, Xiaoli;Park, B. Kevin
氟氯西林是一种用于治疗葡萄球菌感染的合成青霉素。对药物的不良反应被认为是通过蛋白质的共价修饰引起的，组织损伤继发于免疫反应。加合物特异性抗体显示血清蛋白可被氟氯西林修饰，但修饰的性质和位点尚未确定。在这里，在HSA的体外研究表明，通过MS的蛋白质赖氨酸残基的修饰发生在剂量，时间和位点依赖性的方式。亲和，阳离子交换和反相色谱之前，MS显示在耐受性患者中的HSA与氟氯西林的体内修饰，在每个患者中检测到多达9个修饰的赖氨酸残基，并且在8/8例患者中检测到Lys 190和Lys 212的修饰。研究还首次揭示了血浆蛋白可以被氟氯西林的5-羟甲基代谢物修饰，并且母体药物及其代谢物的靶向位点基本相同。本研究提供了内源性靶标化学修饰位点的详细表征，并揭示了用于氟氯西林超敏反应的T细胞和抗体测定的候选肽。
Flucloxacillin is a synthetic penicillin used in the treatment of Staphylococcal infections. Adverse reactions to the drug are believed to arise through covalent modification of proteins, with tissue damage occurring secondary to an immune reaction. Serum proteins have been shown by adduct-specific antibodies to be modified by flucloxacillin, but the nature and sites of modification have not been characterised. Here, in vitro studies on HSA have shown by MS that the modification of protein lysine residues occurs in a dose-, time- and site-dependent manner. Affinity, cation exchange and reversed phase chromatography prior to MS revealed in vivo modification of HSA with flucloxacillin in tolerant patients, with up to nine modified lysine residues being detected in each patient, and with modification of Lys190 and Lys212 being detected in 8/8 patients. It was also revealed for the first time that plasma proteins could be modified with the 5-hydroxymethyl metabolite of flucloxacillin, and that essentially the same Lys residues were targeted by both the parent drug and its metabolite. This study provides a detailed characterisation of sites of chemical modification of an endogenous target and reveals candidate peptides for T-cell and antibody assays of flucloxacillin hypersensitivity.