土鳖虫小肽体外稳定性与安全性评价研究

Research of Stability in Vitro and Safety Assessment on Male Eupolyphaga Sinensis Walker

目的：考察土鳖虫小肽DP-17、LL-8在不同p H、不同温度、体外消化的稳定性,及其在小鼠体内的毒性研究。方法：体外稳定性实验,采用HPLC方法,以TSK-SWG2000WL凝胶柱为分析手段,测定在酸性、中性、碱性条件下DP-17、LL-8的稳定性;同时考察DP-17、LL-8在温度为-20℃、0℃、常温条件、37℃、体外消化下的稳定性,安全性评价实验,采用腹腔注射的方式,以各个组织湿重变化、器官指数、进食量、饮水量、呼吸频率、体重变化为指标,对小鼠进行安全性评价。结果：土鳖小肽DP-17、LL-8在不同p H中出峰时间稳定,峰高无变小趋势,同时在体外消化、不同温度下出峰时间分别为12.5、13 min左右,说明土鳖虫小肽在不同p H、体外消化、不同温度下性质稳定,无分解现象;此外,通过小鼠长期和急性毒性实验,大鼠外观体征未见异常变化,实验组进食量和饮水量与对照组相当,体重和各个组织湿重无明显变化,呼吸频率平稳。结论：土鳖虫小肽在体外的稳定性较好;小鼠安全性评价实验表明,土鳖虫小肽无毒性,安全评价良好,为后期药代动力学研究做基础。

Objective：Study on stability in different p H,temperature,digestion in vitro and toxicity in mice of small peptide DP-17,LL-8 from Eupolyphaga sinensis Walker. Methods：In vitro stability test was carried out by HPLC and TSK-SWG2000 WL gel column as the analysis method,the stability of DP-17 and LL-8 from Eupolyphaga sinensis Walker was determined in acidic,neutral and alkaline conditions. The stability of DP-17 and LL-8 was investigated at the temperature of-20 ℃,0 ℃,normal temperature,37 ℃ and digestion in vitro,the safety evaluation experiment was performed by intraperitoneal injection,the indexes of safety evaluation of mice was evaluated by the changes of wet weight,organ index,food intake,water consumption,respiratory rate and weight change. Results：The peak time and peak height were change tendency on DP-17 and LL-8,at the same time,peak times were 12.5 min,13 min in vitro digestion under different temperature,long term and acute toxicity tests were carried out in mice,the tests showed that the experimental group were equal to the control group on amount of food and water. There was no significant change in body weight and tissue wet weight,and the respiratory rate. Conclusion：Stability of DP-17,LL-8 are good,the evaluation result showed that no toxic of DP-17,LL-8 and safety evaluation is good. It is the basis for the study of pharmacokinetics.