Liquid Chromatography Combined With Tandem Mass Spectrometry for the Pharmacokinetic and Metabolism Studies of PROTAC ARV-471 in Rats

IF 1.8 4区医学 Q4 BIOCHEMICAL RESEARCH METHODS

Biomedical Chromatography Pub Date : 2025-01-02 DOI:10.1002/bmc.6068

Wenbin Huang, Hongjian Zhang, Yayun Zhou, Xuechao Liu, Qilei Zhang, Xiaopeng Ma

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Abstract

Proteolysis targeting chimera (PROTAC) is emerging as a promising medicinal modality, which has aroused widespread interest among the field of pharmaceutical manufacturing in the recent years. ARV-471 is an orally active PROTAC estrogen receptor degrader against breast cancer, which leads to the ubiquitylation and subsequent degradation of estrogen receptors via the proteasome. In this study, we developed a highly sensitive liquid chromatography tandem mass spectrometry method (LLOQ = 0.5 ng/mL) for the measurement of ARV-471 in rat plasma. The acetonitrile precipitated sample was separated on ACQUITY BEH C₁₈ column using acetonitrile-0.1% formic acid as mobile phased with gradient elution. Multiple reactions monitoring in positive ESI mode was employed for the quantification of ARV-471 (m/z 724.4 → 396.2). The assay showed good linearity over the concentration range of 0.5–1000 ng/mL with correlation coefficient > 0.996. The assay was validated according to FDA guidance, and all the validation parameters were within the predefined acceptance criteria. After validation, the assay was applied to the pharmacokinetic study of ARV-471 in rats. Additionally, the metabolites in rat plasma were identified using liquid chromatography–high resolution mass spectrometry. Four metabolites were identified and characterized. Hydrolysis, glucuronidation and deamination were the main metabolic pathways of ARV-471 in rats.

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液相色谱结合串联质谱法研究PROTAC ARV-471在大鼠体内的药动学和代谢。

蛋白水解靶向嵌合体（Proteolysis targeting chimera， PROTAC）是一种新兴的有前景的药物治疗方式，近年来引起了医药制造领域的广泛关注。ARV-471是一种口服活性的抗乳腺癌PROTAC雌激素受体降解剂，它通过蛋白酶体导致雌激素受体的泛素化和随后的降解。本研究建立了一种高灵敏度的液相色谱串联质谱法（LLOQ = 0.5 ng/mL）测定大鼠血浆中ARV-471的方法。采用ACQUITY BEH C18色谱柱，以乙腈-0.1%甲酸为流动相，梯度洗脱，分离乙腈沉淀样品。采用ESI阳性模式多反应监测定量ARV-471 （m/z 724.4→396.2）。在0.5 ~ 1000 ng/mL范围内线性良好，相关系数为> 0.996。根据FDA指南进行验证，所有验证参数均在预定义的验收标准范围内。验证后，将该方法应用于ARV-471在大鼠体内的药代动力学研究。此外，采用液相色谱-高分辨质谱法对大鼠血浆中的代谢物进行了鉴定。鉴定并表征了四种代谢物。水解、糖醛酸和脱胺是ARV-471在大鼠体内的主要代谢途径。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biomedical Chromatography 生物-分析化学

CiteScore

3.60

自引率

5.60%

发文量

268

审稿时长

2.3 months

期刊介绍： Biomedical Chromatography is devoted to the publication of original papers on the applications of chromatography and allied techniques in the biological and medical sciences. Research papers and review articles cover the methods and techniques relevant to the separation, identification and determination of substances in biochemistry, biotechnology, molecular biology, cell biology, clinical chemistry, pharmacology and related disciplines. These include the analysis of body fluids, cells and tissues, purification of biologically important compounds, pharmaco-kinetics and sequencing methods using HPLC, GC, HPLC-MS, TLC, paper chromatography, affinity chromatography, gel filtration, electrophoresis and related techniques.

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