抗人类免疫缺陷病毒先导物DAAN-5508的大鼠药动学研究

Pharmacokinetics Study of the Anti-HIV Lead Compound DAAN-5508 in Rats

目的建立检测大鼠血浆中抗人类免疫缺陷病毒（human immunodeficiency virus，HIV）候选化合物DAAN-5508的LC—MS／MS方法，并将其应用于大鼠体内药动学研究。方法血浆样品用甲醇沉淀蛋白处理；色谱分离用含0．1％甲酸的甲醇和水为流动相梯度洗脱，流速为0．3mL·min^-1。定量分析在Triple Quad LC—MS／MS上采用电喷雾离子化电离源（ESI）、正离子多重反应离子监测（MRM）的方式进行。将建立的方法应用于大鼠静注和口服DAAN-5508的药动学和生物利用度研究。结果DAAN-5508在0．2～2500ng·mL^-1内呈良好的线性关系（r^2=0．9998），最低定量限为0．2ng·mL^-1。方法的回收率〉80％，日内和日间精密度和准确度符合生物样品的检测要求。大鼠静注（5mg·kg^-1）和口服（15mg·kg^-1）DAAN-5508后，静注和口服的消除半衰期分别为2．6和4．6h。大鼠口服后DAAN-5508的吸收较快，血浆浓度在1h左右达到（188．0±62．33）ng·mL^-1的峰值，口服生物利用度为12％。结论本实验建立了特异、灵敏、简便快捷的定量检测血浆DAAN-5508的LC—MS／MS方法，成功应用于DAAN-5508的大鼠药动学和生物利用度研究。

OBJECTIVE To develop and validate a LC-MS/MS method for quantitative analysis of the new anti-14IV candidate DAAN-5508 in rat plasma, and to study its pharmacokinetics and bioavailability in rats. METHODS The plasma samples were treated with methanol for precipitating proteins. The chromatographic separation was performed with a gradient elution using 0. 1% fomuc acid solution and methanol containing 0. 1% formic acid. The flow rate was 0. 3 mL · min^-1. The MS detection was conducted using an LC-MS/MS in positive ion electrospray and multiple reactions monitoring （MRM） mode. RESULTS Good linearity was obtained over the concentration range of 0. 2 - 2 500 ng · mL^-1 for DAAN-5508 （ r^2 = 0. 999 8）, with the lower limit of quantification at 0.2 ng · mL^-1. The recovery of DAAN-5508 was greater than 80%. The precision and the accuracy met the requirements for bioanalysis. The method was applied for pharmacokineties study of DAAN-5508 in rats. After a single iv（5 mg· kg^-1 ）Or oral dose （ 15 mg· kg^-1 ）of DAAN-5508 to rats, the plasma coneentration of DAAN-5508 showed a biphasic decline. The elimination half-lives were 2. 6 and 4. 6 h for intravenous and oral administration, respectively. The absorption of DAAN-5508 was rapid after oral administration. The peak level （ 188.0 ± 62. 33 ） ng · mL^-1 was reached at 1 h. The oral bioavailability was 12%. CONCLUSION The developed the LC-MS/MS method is selective, sensitive, rapid and simple. It is suitable for the in vivo pharmacokinetics study of DAAN-5508 in rats.