Liquid chromatography-tandem mass spectrometry methods for quantification of roxadustat (FG-4592) in human plasma and urine and the applications in two clinical pharmacokinetic studies
Liquid chromatography-tandem mass spectrometry methods for quantification of roxadustat (FG-4592) in human plasma and urine and the applications in two clinical pharmacokinetic studies
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液相色谱-串联质谱法定量人血浆和尿液中罗沙司他（FG-4592）的含量及其在两项临床药代动力学研究中的应用
DOI：
10.1016/j.jchromb.2022.123274
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Pei Hu
中科院分区：
文献类型：
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作者：
Xin Zheng;Xia Chen;Tao Liu;Ji Jiang;Xinge Cui;Qian Zhao;Pei Hu
·建立了两种方法来定量人血浆和尿液中的roxadustat。·在样品收集到分析的过程中对方法进行了验证。·这些方法应用于中国首次人体PK研究中。·这些方法支持Roxadustat在中国的批准。Roxadustat（FG-4592）可抑制缺氧诱导因子脯氨酰羟化酶（HIF-PH），该酶负责靶向和调节HIF的泛素化和蛋白酶体降解。它已在中国被批准为治疗透析依赖性CKD（慢性肾病）患者贫血的抗贫血药物，并正在日本接受监管审查。一些研究也在研究roxadustat在CKD患者中的临床药代动力学和药效学。为了支持临床研究，设计了一种快速高效液相色谱-串联质谱（HPLC-MS/MS）方法，用于定量测定人血浆和尿液中的罗沙度司他，浓度范围为1 - 5000 ng/mL。采用液-液萃取（LLE）净化样品，然后在沃茨XTerra Phenyl柱上进行等度洗脱色谱分离。采用由乙腈/水/甲酸（v/v/v）组成的混合物作为移动的相（流速：1.0 mL/min），具有60%柱后分流。采用基于正离子电喷雾电离（ESI）的多反应监测（MRM）模式进行定量。该方法已在中国健康受试者中进行了充分验证，并应用于roxadustat的药代动力学（PK）和药效学（PD）研究。
• Two methods were established to quantify roxadustat in human plasma and urine. • The methods were validated within the progress of sample-collection to analysis. • These methods were applied in the first-in-human PK study of roxadustat in China. • These methods supported the approval of Roxadustat in China. Roxadustat (FG-4592) can inhibit the hypoxia-inducible factor prolyl hydroxylase (HIF-PH) enzymes that are responsible for targeting and regulating HIF for ubiquitination and proteasomal degradation. It has been approved in China as an anti-anemia drug for treating the anemia in dialysis-dependent CKD (chronic kidney disease) patients, and is also under regulatory review in Japan. Some studies are also investigating the clinical pharmacokinetics and pharmacodynamics of roxadustat in CKD patients. To support clinical investigations, a rapid high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method, with a concentration range from 1 to 5000 ng/mL, was designed for the quantification of roxadustat in human plasma and urine. Liquid-liquid extraction (LLE) was applied to sample clean-up followed by a chromatographic separation conducted on a Waters XTerra Phenyl column with isocratic elution. The mixture consisting of acetonitrile/water/formic acid v/v/v) was employed as the mobile phase (flow rate: 1.0 mL/min) with 60% post-column split. The quantitation was employed in multiple reactions monitoring (MRM) mode based on positive electrospray ionization (ESI). This proposed method was fully validated and applied to the pharmacokinetic (PK) and pharmacodynamic (PD) study of roxadustat among healthy subjects in China.