关节腔注射用雷公藤甲素微球组织分布与代谢

Tissue Distribution and Metabolism of Triptolide Microspheres After Articular Cavity Injection in Rats

目的考察关节腔注射用雷公藤甲素微球(TPL-MS)健康大鼠组织分布与体内外代谢行为。方法建立超高效液相色谱-串联质谱(UPLC-MS/MS)方法,测定不同时间点TPL-MS在大鼠心、肝、脾、肺、肾、血浆、关节的分布;以雷公藤甲素溶液(TPL-IN)为参比,通过测定TPL-MS在大鼠肝匀浆和肝微粒体酶中代谢后的剩余量,计算不同时间点雷公藤甲素代谢百分比;对健康大鼠进行关节腔给药,采用高分辨质谱分析TPL-MS在大鼠肝脏、胆汁、尿液和粪便中的代谢物种类和代谢速度。结果给药后30 min,TPL-IN在肝匀浆和肝微粒体中分别下降50.32%、49.47%,TPL-MS分别下降47.62%、46.36%;体内外共观察到4种代谢物,两种制剂体内代谢途径和代谢物种类相似,TPL-IN主要代谢时间集中在给药后8 h,TPL-MS可在给药后0~24 h持续检出代谢物;TPL-MS给药后第3天在关节组织内达到最高浓度,维持时长接近2周。结论2种制剂体外代谢规律基本一致,体内代谢途径和代谢物种类一致。与TPL-IN相比,TPL-MS的组织分布和代谢速度发生明显改变,关节滞留时间延长,体内代谢过程延缓。

Objective To investigate the in vitro and in vivo metabolic behavior and tissue distribution of triptolide microspheres(TPL-MS)after articular cavity injection on healthy rats.Methods An ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method was established to determine the distribution of TPL-MS in the heart,liver,spleen,lung,kidney,plasma and joints at different time points.Using TPL solution(TPL-IN)as reference,the metabolic percentage of TPL at different time points was calculated by measuring the residual amount of TPL in rat liver homogenate and liver microsomal enzyme after metabolism.Joint cavity administration was performed on healthy rats.Then the metabolites and metabolic rate of TPL-MS in liver,bile,urine and feces were analyzed by high resolution mass spectrometry.Results At 30 min,the contents of TPL-IN in liver homogenate and liver microsomes decreased by 50.32%and 49.47%.The TPL-MS decreased by 47.62%and 46.36%,respectively.Four metabolites were found and the metabolic types of the two preparations were similar in vivo and in vitro.The main metabolic time of the TPL-IN group was concentrated within 8 h after administration,and the metabolites of the TPL-MS group were continuously detected within 0-24 h.The maximum concentration of microspheres was reached in the joint tissue on day 3 and maintained for nearly 2 weeks.Conclusion The metabolic law in vitro,metabolic pathway,and metabolite types in vivo of the two preparations were consistent.Compared with TPL-IN group,the tissue distribution and metabolic rate of TPL-MS were significantly changed,the joint retention time was prolonged,and the metabolic process in vivo was also delayed.