Identification of Human Liver Cytochrome P450 Enzymes Involved in the Metabolism of SCH 530348 (Vorapaxar), a Potent Oral Thrombin Protease-Activated Receptor 1 Antagonist
Identification of Human Liver Cytochrome P450 Enzymes Involved in the Metabolism of SCH 530348 (Vorapaxar), a Potent Oral Thrombin Protease-Activated Receptor 1 Antagonist
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DOI：
10.1124/dmd.110.035493
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发表时间：
2011-01-01
影响因子：
3.9
通讯作者：
Alton, Kevin B.
中科院分区：
文献类型：
--
作者：
Ghosal, Anima;Lu, Xiaowen;Alton, Kevin B.
Vorapaxar (SCH 530348) 是一种有效的口服凝血酶蛋白酶激活受体 1 拮抗剂，目前正在开发作为一种抗血小板药物，用于治疗患有血管疾病的患者。本研究的目的是确定负责 SCH 530348 代谢的人肝细胞色素 P450 (P450) 酶。人肝微粒体将 SCH 530348 代谢为 M19（一种通过氨基甲酸酯裂解形成的胺代谢物）和 M20（单羟基-SCH 530348）。重组人 CYP3A4 对 M19 的形成表现出最高的活性（11.5% 的放射活性），其次是 CYP1A1 和 CYP2C19 的底物转化明显较少。通过 CYP1A2、CYP3A5 和 CYP4F3A 检测到痕量 M19（一种主要的人体排泄代谢物）。人肝微粒体M19的形成被酮康唑抑制89%（IC(50)，0.73μM），反苯环丙明抑制34%，抗CYP3A4单克隆抗体抑制89%。 M19 形成率与咪达唑仑 1'-羟基化率 (r = 0.75) 或 M19 形成率与睾酮 6 β-羟基化率 (r = 0.92) 之间存在显着相关性。筛选、抑制和相关性研究的结果证实，CYP3A4 是负责从 SCH 530348 形成 M19 的主要 P450 酶。相比之下，稳态下主要循环人类代谢物 M20 的形成主要由 CYP3A4 和 CYP2J2 催化。 M20 在药理学上与 SCH 530348 等效，而 M19 是一种无活性的代谢物。人肝微粒体 M20 的形成被酮康唑抑制 89%，阿司咪唑（CYP2J2 抑制剂）抑制 75%，CYP3A4 单克隆抗体抑制 43%。这些结果表明 CYP3A4 和 CYP2J2 均参与 M20 代谢物的形成。
Vorapaxar (SCH 530348), a potent oral thrombin protease-activated receptor 1 antagonist, is being developed as an antiplatelet agent for patients with established vascular disease. The objective of this study was to identify the human liver cytochrome P450 (P450) enzyme(s) responsible for the metabolism of SCH 530348. Human liver microsomes metabolized SCH 530348 to M19, an amine metabolite formed via carbamate cleavage, and M20 (monohydroxy-SCH 530348). Recombinant human CYP3A4 exhibited the most activity (11.5% profiled radioactivity) for the formation of M19, followed by markedly less substrate conversion with CYP1A1 and CYP2C19. Trace levels of M19, a major excreted human metabolite, were detected with CYP1A2, CYP3A5, and CYP4F3A. Formation of M19 by human liver microsomes was inhibited 89% by ketoconazole (IC(50), 0.73 mu M), 34% by tranylcypromine, and 89% by anti-CYP3A4 monoclonal antibody. There was a significant correlation between the rate of M19 formation and midazolam 1'-hydroxylation (r = 0.75) or M19 formation and testosterone 6 beta-hydroxylation (r = 0.92). The results of screening, inhibition, and correlation studies confirmed that CYP3A4 is the major P450 enzyme responsible for M19 formation from SCH 530348. In contrast, formation of M20, a major circulating human metabolite at steady state, was primarily catalyzed by CYP3A4 and CYP2J2. M20 is pharmacologically equipotent to SCH 530348, whereas M19 is an inactive metabolite. Formation of M20 by human liver microsomes was inhibited 89% by ketoconazole, 75% by astemizole (a CYP2J2 inhibitor), and 43% by CYP3A4 monoclonal antibody. These results suggest that CYP3A4 and CYP2J2 are both involved in the formation of M20 metabolite.