Quantification of therapeutic miRNA mimics in whole blood from nonhuman primates.
Quantification of therapeutic miRNA mimics in whole blood from nonhuman primates.
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DOI：
10.1021/ac403044t
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发表时间：
2014-02-04
影响因子：
7.4
通讯作者：
Bader, Andreas G.
中科院分区：
文献类型：
--
作者：
Kelnar, Kevin;Peltier, Heidi J.;Leatherbury, Neil;Stoudemire, Jay;Bader, Andreas G.
MRX34是一种基于microRNA（miRNA）的癌症疗法，最近已进入临床试验，成为同类产品中的第一个临床候选药物。它是一种脂质体纳米颗粒，负载有肿瘤抑制剂miRNA miR-34a的合成模拟物作为活性药物成分。为了了解药物的药代动力学特性并使临床中的最佳给药方案合理化，需要一种定量检测miRNA模拟物的方法。在这里，我们报告了定量逆转录-聚合酶链反应（qRT-PCR）测定的开发和鉴定，以支持非人灵长类动物的药代动力学和毒代动力学评估。对从全血中分离的总核糖核酸（RNA）进行检测和定量。标准曲线的合格范围跨越6个数量级，从2.5 × 10 - 7至2.5 × 10 - 1 ng/逆转录（RT）反应，对应于6.2 × 10 - 5至6.2 × 101 ng/mL的估计血药浓度。我们的结果表明，内源性以及外源性miR-34a可以准确和精确地定量。该试验用于建立MRX34的药代动力学特征，显示miRNA模拟物在非人灵长类动物全血中的有利停留时间和暴露量。
MRX34, a microRNA (miRNA)-based therapy for cancer, has recently entered clinical trials as the first clinical candidate in its class. It is a liposomal nanoparticle loaded with a synthetic mimic of the tumor suppressor miRNA miR-34a as the active pharmaceutical ingredient. To understand the pharmacokinetic properties of the drug and to rationalize an optimal dosing regimen in the clinic, a method is needed to quantitatively detect the miRNA mimic. Here, we report the development and qualification of a quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay in support of pharmacokinetic and toxicokinetic assessments in the nonhuman primate. Detection and quantification were performed on total ribonucleic acid (RNA) isolated from whole blood. The qualified range of the standard curve spans 6 orders of magnitude from 2.5 × 10–7 to 2.5 × 10–1 ng per reverse transcription (RT) reaction, corresponding to an estimated blood concentration from 6.2 × 10–5 to 6.2 × 101 ng/mL. Our results demonstrate that endogenous as well as the exogenous miR-34a can be accurately and precisely quantified. The assay was used to establish the pharmacokinetic profile of MRX34, showing a favorable residence time and exposure of the miRNA mimic in whole blood from nonhuman primates.
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影响因子：
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