Wang Jiang, Zhu Xiaohua, Kolli Shamala, Wang Hongyan, Pearce Cedric J, Oberlies Nicholas H, Phelps Mitch A
Comprehensive Cancer Center, The Ohio State University, Columbus, OH, United States.
College of Pharmacy, The Ohio State University, Columbus, OH, United States.
J Pharm Biomed Anal. 2017 May 30;139:187-192. doi: 10.1016/j.jpba.2017.02.051. Epub 2017 Mar 1.
Verticillin A is a natural product isolated from fungal cultures and has displayed potent antibiotic, antiviral, nematocidal, and anticancer properties in vitro. While in vivo studies have been limited due to sparse supply, the in vivo efficacy data that does exist demonstrates potent anti-tumor activity in murine cancer models. The current study aims to investigate the pharmacokinetics and bioavailability of verticillin A in mice to provide guidance for further efficacy assessment in mouse models. A sensitive and specific liquid chromatography-tandem mass spectrometry method was developed and validated for the quantification of verticillin A in mouse plasma. Sample preparation was accomplished through protein precipitation, and chromatographic separation was achieved on an Agilent Zorbax Extend C18 column with a security guard cartridge C8 using a binary gradient with mobile phase A (water/0.1% formic acid) and B (ACN/0.1% formic acid) at a flow rate of 400μl/min. Elution of verticillin A and internal standard, hesperetin, occurred at 4.87 and 2.06min, respectively. The total chromatographic run time was 8min, and the assay was linear in the concentration range of 1-1000nM. The within- and between day precisions and accuracy were in the range of 2.58-8.71 and 90-105%, respectively. The assay was applied to determine plasma drug concentration in a mouse pharmacokinetic study. It was found that intraperitoneal dosing of 3mg/kg resulted in high systemic exposure and achieved C of 110nM with plasma concentrations sustained above 10nM for the 24-h duration of the study. Intravenous and oral dosing achieved observed C of 73nM and 9nM, respectively. Oral dosing resulted in an approximate 9% bioavailability. Comparing with previously published in vitro studies that demonstrated verticillin A is active in the 20nM to 130nM range, the pharmacokinetic data demonstrate similar levels are achieved in mouse plasma via intravenous or intraperitoneal dosing routes.
弗替西林A是一种从真菌培养物中分离出的天然产物，在体外已显示出强效的抗生素、抗病毒、杀线虫和抗癌特性。虽然由于供应稀少，体内研究受到限制，但现有的体内疗效数据表明，其在小鼠癌症模型中具有强效的抗肿瘤活性。本研究旨在研究弗替西林A在小鼠体内的药代动力学和生物利用度，为在小鼠模型中进一步评估疗效提供指导。开发并验证了一种灵敏且特异的液相色谱-串联质谱法，用于定量小鼠血浆中的弗替西林A。样品制备通过蛋白沉淀完成，色谱分离在安捷伦Zorbax Extend C18柱上进行，配备保护柱C8，使用二元梯度，流动相A（水/0.1%甲酸）和B（乙腈/0.1%甲酸），流速为400μl/min。弗替西林A和内标橙皮素的洗脱时间分别为4.87分钟和2.06分钟。总色谱运行时间为8分钟，该测定法在1-1000nM的浓度范围内呈线性。日内和日间精密度及准确度分别在2.58-8.71和90-105%的范围内。该测定法应用于小鼠药代动力学研究中血浆药物浓度的测定。结果发现，腹腔注射3mg/kg导致全身暴露水平较高，在研究的24小时内达到110nM的Cmax，血浆浓度持续高于10nM。静脉注射和口服给药的Cmax分别为73nM和9nM。口服给药的生物利用度约为9%。与先前发表的体外研究表明弗替西林A在20nM至130nM范围内具有活性相比，药代动力学数据表明，通过静脉或腹腔给药途径在小鼠血浆中可达到相似的水平。
