阿德福韦衍生物灌胃给药后的代谢产物阿德福韦在大鼠体内的药动学研究

Study on Pharmacokinetics of Metabolites of Adefovir in Rats after Intragastric Administration of Adefovir Derivatives

目的:建立阿德福韦(PMEA)的含量测定方法,并研究阿德福韦衍生物[阿德福韦前药,阿德福韦-熊去氧胆酸-3-丙基酯,L-亮氨酸-3-丙基酯(PMEA-1c)]灌胃给药后的代谢产物PMEA在大鼠体内的药动学。方法:采用高效液相色谱-串联质谱(HPLCMS/MS)法。色谱柱为BEH C_(18),流动相为0.1%甲酸乙腈-0.1%甲酸水(梯度洗脱),流速为0.25 mL/min,柱温为30℃,进样量为1μL;检测离子对为PMEA质荷比(m/z)274.1→162.1,葛根素(内标)m/z 417.1→267.1。12只大鼠随机分为阿德福韦酯(ADV)组(阳性对照,90 mg/kg)、PMEA-1c组(160 mg/kg),每组6只,分别灌胃给予相应药物1次,于给药后0.083、0.25、0.5、0.75、1、2、4、6、8、10、12、24 h尾静脉取血测定PMEA血药浓度;利用DAS 2.0软件计算相关药动学参数。结果:PMEA检测质量浓度线性范围为6.1～440.0 ng/mL(r=0.998 5),日内、日间精密度和稳定性试验的RSD均<10%(n=3、5、6),准确度为82.16%～97.33%(RSD≤6.4%,n=5),基质效应为95.96%～106.35%(RSD≤4.9%,n=5);ADV组和PMEA-1c组大鼠血浆中PMEA的t_(1/2)分别为(1.762±0.117)、(2.548±0.174)h,AUC_(0-24) h分别为(2 170.059±146.091)、(4 704.257±176.792)μg·h/L,c_(max)分别为(613.092±9.504)、(697.295±15.275)μg/L;与ADV组比较,PMEA-1c组大鼠t_(1/2)、AUC_(0-24 h)、AUC_(0-∞)、c_(max)均显著增加(P<0.01或P<0.001)。结论:建立的方法准确可靠,本实验结果表明PMEA-1c代谢为单室模型,其可作为阿德福韦前药的潜力药物,为阿德福韦前药的继续研究奠定了基础。

OBJECTIVE:To establish a method for the determination of adefovir(PMEA)and study the pharmacokinetics of metabolites PMEA in rats after intragastric administration of PMEA derivatives [PMEA prodrug, adefovir-ursodeoxycholic acid-3-propyl ester, L-leucine-3-propyl ester (PMEA-1c)] in rats. METHODS: HPLC-MS/MS method was adopted. The determination was performed on BEH C18 column with mobile phase consisted of 0.1% formic acid acetonitrile-0.1% formic acid water(gradient elution)at the flow rate of 0.25 mL/min. The column temperature was 30 ℃,and sample size was 1 μL. The quantitative ions were PMEA m/z 274.1→162.1,puerarin(internal standard)m/z 417.1→267.1. 12 rats were randomly divided into adefovir dipivoxil(ADV)group(positive control,90 mg/kg)and PMEA-1c group(160 mg/kg),with 6 rats in each group. They were given relevant medicine once intragstrically,and the blood samples were collected from tail vein 0.083,0.25,0.5,0.75,1, 2,4,6,8,10,12,24 h after administration to determine the plasma concentration of PMEA. Relevant pharmacokinetic parameters were calculated by using DAS 2.0 software. RESULTS:The linear range of PMEA was 6.1-440.0 ng/mL(r=0.998 5). RSDs of intra and inter day of precision and stability tests were all less than 10%(n=3,5,6),and the accuracy was 82.16%-97.33%(RSD≤6.4%,n=5). Matrix effects ranged from 95.96%-106.35%(RSD≤4.9%,n=5). The pharmacokinetic parameters of PMEA in ADV group and PMEA-1c group were as follows as t1/2 were(1.762±0.117)and(2.548±0.174)h;AUC0-24 h were (2 170.059 ± 146.091) and (4 704.257 ± 176.792)μg·h/L;cmax were(613.092±9.504)and(697.295± 15.275)μg/L,respectively. Compared with ADV group,t1/2, AUC0-24 h,AUC0-∞ and cmax of rats in PMEA-1c group were increased significantly(P<0.01 or P<0.001). CONCLUSIONS:Established method is accurate and reliable. The trial indicates that PMEA-1c metabolism is single compartment model,show that can be used as a potential prodrug for adefovir,which lay a foundation for the further study of adefovir prodrug.