雷公藤内酯醇长循环纳米脂质体的急性毒性和长期毒性研究

Acute toxicity and long-term toxicity study of long-circulating nanoliposomes of triptolide

目的考察多肽修饰的雷公藤内酯醇长循环纳米脂质体(PTN)的毒性特点,为临床前安全性评价提供依据。方法80只健康ICR小鼠按完全随机分组法分为8组,每组各10只,设置正常组、阴性对照组、雷公藤内酯醇(TP)阳性对照组和5个PTN剂量组,采用单次最大药量法,尾静脉注射给药,观察给药后小鼠行为、体重、毛色等外观变化,记录小鼠死亡数量,计算LD_(50)。按照等效剂量系数换算大鼠的LD_(50),设计1/10 LD_(50)、1/50 LD_(50)、1/100 LD_(50)三个剂量进行长期毒性实验;将32只健康SD大鼠按完全随机分组法分成8组,每组各4只,设置正常组,阴性对照组,阳性对照药TP及PTN高、中、低剂量组,采用尾静脉注射方式每2天给药一次,连续给药28天。期间观察动物的行为、毛色光泽、排泄物、摄食量等一般情况,每7天称重一次,停药12 h后,检测动物脏器重量与系数、血清生化指标。结果由急性毒性实验可知,PTN对小鼠的LD_(50)为1.158 mg/kg,95%的置信区间为0.876~1.161 mg/kg。长期毒性实验,PTN对大鼠的摄食量、体重、脏器重量和脏器系数变化影响,差异无统计学意义(P>0.05);与正常组比较,TP对照和PTN实验组大鼠血清生化检查指标谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、血尿素氮(BUN)值均升高,白蛋白(ALB)、血清白/球蛋白(A/G)、谷草转氨酶/谷丙转氨酶(AST/ALT)值降低,其中TP高、中、低剂量组的生化指标变化比较,差异有统计学意义(P<0.05),PTN高、中、低剂量组的变化比较,差异无统计学意义(P>0.05)。结论PTN给药系统尾静脉注射能够有效降低TP的毒性。

Objective To investigate the toxicity characteristics of peptide-modified long-circulating nanoliposomes(pleiotrophin, PTN) of triptolide(TP), so as to provide a basis for preclinical safety evaluation.Methods 80 healthy ICR mice were divided into a normal group, a negative control group, a positive control TP group, and 5 PTN dose groups according to the complete random grouping method, with 10 mice in each group. The drug was injected via tail vein using the single maximum dose method. The mice were observed for changes in behavior, body weight, hair color and other appearances after administration, and the number of mouse deaths was recorded to calculate the median lethal dose(LD_(50)). The LD_(50)of rats was converted according to the equivalent dose coefficient, and three doses of 1/10 LD_(50), 1/50 LD_(50)and 1/100 LD_(50)were designed for long-term toxicity tests. 32 healthy SD rats were divided into a normal group, a negative control group, positive control TP and PTN high, medium and low dose groups according to the complete randomization method, with 4rats in each group. The drug was injected via tail vein once every 2 days for 28 d. During the injection period,the rats were observed for their general conditions such as behavior, hair color and luster, excretion and food intake, and weighed every 7 d. After 12 h of drug discontinuation, animal organ weights and coefficients, as well as serum biochemical indicators were examined. Results The acute toxicity test results showed that the LD_(50)of PTN on mice was 1.158 mg/kg with 95% confidence interval of 0.876-1.161 mg/kg. The long-term toxicity test results showed no statistical significances between PTN and the changes of food intake, body weight, organ weight and organ coefficient of rats(P > 0.05). Compared with the normal group, the values of serum biochemical indicators, such as alanine transaminase(ALT), aspartate transaminase(AST), alkaline phosphatase(ALP) and blood urea nitrogen(BUN), increased, while the values of albumin(ALB), serum albumin/globulin(A/G) and AST/ALT decreased in the TP control and PTN test groups. There were statistically significant differences in the changes of biochemical indicators among the TP high, medium and low dose groups(P < 0.05), while there were no statistically significant differences in the changes of biochemical indicators among the PTN high, medium and low dose groups(P > 0.05). Conclusion PTN administration system can effectively reduce the toxicity of TP by tail vein injection.