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.

与人类乳腺癌抗性蛋白（hBCRP）的相互作用会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5'-三磷酸腺苷（ATP）水解获得的能量进行底物的跨膜运动，同时释放5'-二磷酸腺苷（ADP）和无机磷酸盐。ADP解放可用于检测与hBCRP ATPase的相互作用。开发了一种基于亲水相互作用液相色谱（HILIC）结合高分辨率串联质谱（HR-MS / MS）的ADP定量方法，并已根据欧洲药品管理局的生物分析方法验证指南的标准成功进行了验证。定性地包括ATP和5'-单磷酸腺苷，以防止干扰。此外，改进了由六个样品组组成的设置，该设置允许检测hBCRP底物或受试化合物的抑制剂特性。将hBCRP底物柳氮磺吡啶和hBCRP抑制剂原钒酸盐用作对照。为了证明该方法的适用性，测试了氨普那韦，茚地那韦，奈非那韦，利托那韦和沙奎那韦对hBCRP ATPase活性的影响。奈非那韦，利托那韦和沙奎那韦被鉴定为hBCRP ATPase抑制剂，而五种HIV蛋白酶抑制剂中没有一个是hBCRP底物。这些发现与以前的出版物一致。改进了由六个样品组组成的设置，可以检测hBCRP底物或受试化合物的抑制剂特性。将hBCRP底物柳氮磺吡啶和hBCRP抑制剂原钒酸盐用作对照。为了证明该方法的适用性，测试了氨普那韦，茚地那韦，奈非那韦，利托那韦和沙奎那韦对hBCRP ATPase活性的影响。奈非那韦，利托那韦和沙奎那韦被鉴定为hBCRP ATPase抑制剂，而五种HIV蛋白酶抑制剂中没有一个是hBCRP底物。这些发现与以前的出版物一致。改进了由六个样品组组成的设置，可以检测hBCRP底物或受试化合物的抑制剂特性。将hBCRP底物柳氮磺吡啶和hBCRP抑制剂原钒酸盐用作对照。为了证明该方法的适用性，测试了氨普那韦，茚地那韦，奈非那韦，利托那韦和沙奎那韦对hBCRP ATPase活性的影响。奈非那韦，利托那韦和沙奎那韦被鉴定为hBCRP ATPase抑制剂，而五种HIV蛋白酶抑制剂中没有一个是hBCRP底物。这些发现与以前的出版物一致。并测试了沙奎那韦对hBCRP ATPase活性的影响。奈非那韦，利托那韦和沙奎那韦被鉴定为hBCRP ATPase抑制剂，而五种HIV蛋白酶抑制剂中没有一个是hBCRP底物。这些发现与以前的出版物一致。并测试了沙奎那韦对hBCRP ATPase活性的影响。奈非那韦，利托那韦和沙奎那韦被鉴定为hBCRP ATPase抑制剂，而五种HIV蛋白酶抑制剂中没有一个是hBCRP底物。这些发现与以前的出版物一致。