Laboratory of Food Function Analysis, Graduate School of Agricultural Science, Tohoku University, Sendai 980-8572, Miyagi, Japan.
Drug Metabolism and Pharmacokinetics Research Laboratories, Central Pharmaceutical Research Institute, Japan Tobacco Inc., Takatsuki 569-1125, Osaka, Japan.
Molecules. 2023 Feb 7;28(4):1576. doi: 10.3390/molecules28041576.
This study examines 4β-Hydroxycholesterol (4β-HC), which is considered to be a potential marker for the CYP3A4 induction of new chemical entities (NCEs) in drug development. To ensure the use of 4β-HC as a practical biomarker, it is necessary to accurately measure 4β-HC and demonstrate that CYP3A4 induction can be appropriately assessed, even for weak inducers. In clinical trials of NCEs, plasma is often collected with various anticoagulants, in some cases, the plasma is acidified, then stored for an extended period. In this study, we examined the effects of these manipulations on the measurement of 4β-HC, and based on the results, we optimized the plasma collection and storage protocols. We also found that a cholesterol oxidation product is formed when plasma is stored, and by monitoring the compound, we were able to identify when plasma was stored inappropriately. After evaluating the above, clinical drug-drug interaction (DDI) studies were conducted using two NCEs (novel retinoid-related orphan receptor γ antagonists). The weak CYP3A4 induction by the NCEs (which were determined based on a slight decline in the systemic exposure of a probe substrate (midazolam)), was detected by the significant increase in 4β-HC levels (more specifically, 4β-HC/total cholesterol ratios). Our new approach, based on monitoring a cholesterol oxidation product to identify plasma that is stored inappropriately, allowed for the accurate measurement of 4β-HC, and thus, it enabled the evaluation of weak CYP3A4 inducers in clinical studies without using a probe substrate.
本研究考察了 4β-羟基胆固醇（4β-HC），它被认为是药物开发中新化学实体（NCE）CYP3A4 诱导的潜在标志物。为确保将 4β-HC 用作实用的生物标志物，有必要准确测量 4β-HC，并证明即使是弱诱导剂，CYP3A4 的诱导也可以得到适当评估。在 NCE 的临床试验中，通常使用各种抗凝剂采集血浆，在某些情况下，将血浆酸化，然后储存一段时间。在这项研究中，我们检查了这些操作对 4β-HC 测量的影响，并根据结果优化了血浆采集和储存方案。我们还发现，当储存血浆时会形成一种胆固醇氧化产物，通过监测该化合物，我们能够确定何时不当储存了血浆。在评估了上述内容之后，使用两种 NCE（新型视黄醇相关孤儿受体 γ 拮抗剂）进行了临床药物相互作用（DDI）研究。NCE 对 CYP3A4 的弱诱导作用（根据探针底物（咪达唑仑）全身暴露略有下降来确定）通过 4β-HC 水平的显著增加（更具体地说，4β-HC/总胆固醇比值）来检测。我们的新方法基于监测胆固醇氧化产物以识别不当储存的血浆，从而能够准确测量 4β-HC，因此，它能够在不使用探针底物的情况下评估临床研究中的弱 CYP3A4 诱导剂。
