An easy and fast adenosine 5'-diphosphate quantification procedure based on hydrophilic interaction liquid chromatography-high resolution tandem mass spectrometry for determination of the in vitro adenosine 5'-triphosphatase activity of the human breast cancer resistance protein ABCG2.
An easy and fast adenosine 5'-diphosphate quantification procedure based on hydrophilic interaction liquid chromatography-high resolution tandem mass spectrometry for determination of the in vitro adenosine 5'-triphosphatase activity of the human breast cancer resistance protein ABCG2.
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DOI：
10.1016/j.chroma.2017.09.034
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发表时间：
2017-10
期刊：
影响因子：
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通讯作者：
Lea Wagmann;H. Maurer;M. Meyer
中科院分区：
文献类型：
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作者：
Lea Wagmann;H. Maurer;M. Meyer
与人乳腺癌耐药蛋白（hBCRP）的相互作用显着影响药物的药代动力学特性，甚至可能导致药物相互作用。作为ABC超家族的外排泵，hBCRP利用腺苷5′-三磷酸（ATP）水解获得的能量进行底物的跨膜运动，同时释放腺苷5′-二磷酸（ADP）和无机磷酸盐。ADP释放可用于检测与hBCRP ATP酶的相互作用。根据欧洲药品管理局生物分析方法验证指导原则的标准，开发了基于亲水相互作用液相色谱（HILIC）耦合高分辨率串联质谱（HR-MS/MS）的ADP定量方法，并成功验证。定性包括ATP和腺苷5′-一磷酸，以防止干扰。此外，开发了由6个样品组组成的装置，允许检测供试化合物的hBCRP底物或抑制剂特性。hBCRP底物柳氮磺胺吡啶和hBCRP抑制剂原钒酸盐用作对照。为了证明该程序的适用性，检测了安普那韦、茚地那韦、奈非那韦、利托那韦和沙奎那韦对hBCRP ATP酶活性的影响。奈非那韦、利托那韦和沙奎那韦被鉴定为hBCRP ATP酶抑制剂，5种HIV蛋白酶抑制剂均不是hBCRP底物。这些发现与先前的出版物一致。
Interactions with the human breast cancer resistance protein (hBCRP) significantly influence the pharmacokinetic properties of a drug and can even lead to drug-drug interactions. As efflux pump from the ABC superfamily, hBCRP utilized energy gained by adenosine 5′-triphosphate (ATP) hydrolysis for the transmembrane movement of its substrates, while adenosine 5′-diphosphate (ADP) and inorganic phosphate were released. The ADP liberation can be used to detect interactions with the hBCRP ATPase. An ADP quantification method based on hydrophilic interaction liquid chromatography (HILIC) coupled to high resolution tandem mass spectrometry (HR-MS/MS) was developed and successfully validated in accordance to the criteria of the guideline on bioanalytical method validation by the European Medicines Agency. ATP and adenosine 5′-monophosphate were qualitatively included to prevent interferences. Furthermore, a setup consisting of six sample sets was evolved that allowed detection of hBCRP substrate or inhibitor properties of the test compound. The hBCRP substrate sulfasalazine and the hBCRP inhibitor orthovanadate were used as controls. To prove the applicability of the procedure, the effect of amprenavir, indinavir, nelfinavir, ritonavir, and saquinavir on the hBCRP ATPase activity was tested. Nelfinavir, ritonavir, and saquinavir were identified as hBCRP ATPase inhibitors and none of the five HIV protease inhibitors turned out to be an hBCRP substrate. These findings were in line with a pervious publication.