抑制TIMP-1的硫代反义寡核苷酸在肝纤维化大鼠体内生物学分布及药代动力学研究

Study on the chorology and pharmcokinetics of TIMP-1 inhibitor S-asON in rats with hepatic fibrosis

目的 研究抑制TIMP 1的硫代反义寡核苷酸 (S asON )在肝纤维化大鼠体内的生物学分布及药代动力学 ,为其在临床上应用提供理论依据。方法 合成互补于TIMP 1基因的反义寡核苷酸并加以硫代磷酸化修饰 ;用γ 3 2p ATP标记 2 0聚S asON ,给实验大鼠尾静脉一次性注射 ,每 2 4h收集尿液和粪便 ;分别在 12个时间点上放血杀死实验大鼠 ,取各器官组织匀浆 ,用液体闪烁计数测定cpm值。结果 S asON广泛分布于全身各器官 ,以肝、肾聚积浓度最高 ,其药物能在体内维持 72h以上。血浆药物清除曲线符合二室模型。单位时间内机体清除药物的能力即血浆清除率为0 .0 7ml/h ,尿液为主要排除途径 ,2 4h排出 2 8.4% ,72h排出 44 .8% ,粪便排出量较少。结论 抑制TIMP 1的S asON在实验大鼠体内以肝、肾为主的广泛组织生物学分布和较长的清除半衰期 ,使其成药及临床用药变为可能 ,从而为研制新一代抗肝纤维化基因治疗药物奠定了实验基础。肝脏作为反义治疗的靶器官有着重要的现实意义。

Objective To study the biologic distribution and pharmcokinetics of sulfurized thiophosphate modified antisense oligonucleotide (S asON) in rats with hepatic fibrosis, and to provide the theoretical basis for clinical application. Methods Synthesized S asON of TIMP 1, by injecting 20 S asON labelled 5′ end with γ 32p ATP into the tail vein of rats, collected urine and stool per 24h, then killed the exprimental rats at every 12 time points, took out the organs and tissues, and determined the cpm value with liquid scintillometer counter. Results S asON was found to distribute in all organs of the body, with the liver and kidney being the highest, and could be maintained for over 72h. The plasma drug elimination curve was concordant with a two compartment model. Plasma clearance rate was 0.07% /h. Urine was the major clearance way (24h, 28.4% ,72h, 44.8% , respectively). Conclusion In rats model, the liver and kidney are the major histobiologic distributed organs of S asON. Due to its longer clearance half life, it is possible to be used in the clinic practice, the result lays the groundwork for the development of a new gene therapy drug of anti hepatic fibrosis.