注射用多西他赛纳米粒在犬体内的药动学研究

Pharmacokinetics of nanoparticle-docetaxel injection in Beagle dogs

目的建立高通量测定犬血浆中多西他赛浓度的液相色谱-串联质谱(LC-MS/MS)方法,并将其应用于注射用多西他寒纳米粒与多西他赛注射液的药动学比较研究。方法2组Beagle犬分别前肢静脉注射5 mg·kg^(-1)的注射用多西他赛纳米粒和多西他赛注射液后,后肢静脉取血约0.5 mL。血浆样品及内标紫杉醇置于96孔板中,采用8道移液器取液进行高通量的液液萃取。采用LC-MS/MS法测定血药浓度,计算主要药动学参数。结果多西他赛的线性范围为1～500μg·L^(-1),最低定量限为1μg·L^(-1),提取回收率均大于95%,批内、批问精密度(RSD)均小于15%。注射用多西他赛纳米粒和多西他赛注射液,t_(1/2)分别为(5.3±s 1.4)和(3.2±1.4)h,MRT分别为(3.2±1.3)和(0.66±0.21)h,V_(ss)分别为(83±48)和(15±6)L,且2种制剂的t_(1/2)、MRT和V_(ss)均具有显著差异。结论该方法灵敏、准确、专一、简便并且实现高通量分析,适用于多西他赛的药动学比较研究。与多西他赛注射液相比,注射用多西他赛纳米粒能控制药物释放速率,并具有较长的体内循环时间,在一定程度上控制了急性毒性,提高机体耐受性。

AIM To develop a high-throughput analytical method based on LC-MS/MS for the determination of docetaxel and investigate the pharmacokinetics of the two docetaxel preparations in Beagle dog plasma. METHODS Docetaxel and paclitaxel （internal standard, IS） were isolated from 50 μL plasma samples in 96- well plates by liquid-liquid extraction, and most liquid transfer steps were performed by eight-channel pipetting tools. The concentrations of docetaxel in plasma were determined by LC-MS/MS method and the pharmacokinetic parameters were calculated. RESULTS The calibration curves were linear over the range of 1-500 μg·L^-1. The lower limit of quantitation was 1 μg·L^-1. The intra-run and inter-run relative standard deviation （RSD） was less than 15 %. The main pharmacokinetic parameters of nanoparticle-docetaxel injection and docetaxle normal injection were as follows： t1/2 were （5.3±s1.4） and （3.2±1.4） h, MRT were （3.2±1.3） and （0.66±0.21） h, Vss were （83±48） and （15±6） L, respectively. The differences of t1/2, MRT and Vss were significant between two preparations. CONCLUSION The method is sensitive, accurate, convenient and high-throughput for the pharmacokinetic study of nanoparticle-docetaxel injections. Compared with docetaxel normal injection, nanoparticle-docetaxel injection has a controlled releasing rate, a high level of blood concentrations and a long blood circulation time, which are benefit for the control of acute toxicity and the improvement of tolerance to a certain extent.