Dipartimento di Scienze Biomediche e Cliniche "Luigi Sacco", Università degli Studi di Milano, Via G.B. Grassi 74, 20157 Milano, Italy.
Dipartimento di Scienze Biomediche, Chirurgiche ed Odontoiatriche, Sezione di Tossicologia Forense, Università degli Studi di Milano, Via Mangiagalli 37, 20133 Milano, Italy.
Molecules. 2019 Mar 11;24(5):989. doi: 10.3390/molecules24050989.
Olaparib, an orally active inhibitor of poly(ADP-ribose)polymerase(PARP), is the drug of choice in the treatment of gBRCA1/2+ metastatic breast cancers. Unfortunately, Olaparib is poorly soluble with low bioavailability and tumor accumulation; nano-delivery could be a good choice to overcome these disadvantages. Here, a rapid and robust HPLC-ESI⁻MS/MS method for the quantification of Olaparib in ferritin nano-carriers led to the development of cells compartments, different tissues, plasma and urines and were validated to assess the effects of nano-delivery on cell compartment distribution of the drug. This method allows the quantification of Olaparib within the linear range of 0.1⁻10ng/mL in cells culture medium and cell cytoplasm, of 0.5⁻10ng/mL in nuclei, of 0.5⁻100ng/mL in plasma and urine and of 10⁻500ng/mL in tissue samples (kidney and liver). The limit of quantification was found to be 1.54 ng/mL for liver, 2.87 ng/mL for kidney, and lower than 0.48 ng/mL for all matrices. The method has been applied to quantify Ola encapsulated in ferritin-nano-carriers during the nano-drug development. The application of the method to human BRCA-mutated cell model to quantify the Olaparib distribution after incubation of free or ferritin-encapsulated Olaparib is also reported. This sensitive method allows the quantification of low concentrations of Olaparib released from nano-carriers in different cell compartments, leading to the determination of the drug release and kinetic profile of an essential parameter to validate nano-carriers.
奥拉帕利(一种口服聚(ADP-核糖)聚合酶(PARP)抑制剂)是治疗 gBRCA1/2+转移性乳腺癌的首选药物。不幸的是,奥拉帕利的水溶性差,生物利用度低,肿瘤蓄积率低;纳米递药系统可能是克服这些缺点的一个很好的选择。本研究建立了一种快速、稳健的 HPLC-ESI⁻MS/MS 法,用于定量检测载脂蛋白纳米载体中的奥拉帕利,可用于细胞区室、不同组织、血浆和尿液的分析,并验证了该方法在评估纳米递药对药物细胞区室分布的影响。该方法可在细胞培养液和细胞质中 0.1⁻10ng/mL、核中 0.5⁻10ng/mL、血浆和尿液中 0.5⁻100ng/mL、组织样本(肾脏和肝脏)中 10⁻500ng/mL 的线性范围内定量检测奥拉帕利。定量下限(LLOQ)分别为肝脏 1.54ng/mL、肾脏 2.87ng/mL,所有基质的LLOQ均低于 0.48ng/mL。该方法已应用于定量载脂蛋白纳米载体中封装的奥拉帕利,在纳米药物开发过程中。还报道了该方法在人 BRCA 突变细胞模型中的应用,用于定量检测游离或载脂蛋白纳米载体中封装的奥拉帕利孵育后奥拉帕利的分布。该灵敏的方法可以定量检测不同细胞区室中从纳米载体中释放的低浓度奥拉帕利,从而确定药物释放和动力学特征,这是验证纳米载体的关键参数。