霍乱毒素B亚基与人胰岛素B链融合蛋白的安全性评价

Safety evaluation of cholera toxin B subunit-insulin B chain fusion protein

目的评价霍乱毒素B亚基与人胰岛素B链融合蛋白(cholera toxin B subunit-insulin B chain,CTB-INS)在恒河猴体内的安全性。方法将恒河猴随机分为4组,每组10只,雌雄各半,高、中、低剂量组分别经口给予4.0、2.0和1.0 g/(kg·d)CTB-INS,对照组经口给予10 ml/(kg·d)0.5%羧甲基纤维素钠(carboxymethyl cellulose sodium,CMC-Na),每天上午同一时间给药1次,每周给药6 d,连续给药9.0个月。每天上午观察、记录恒河猴的一般体征、摄食量,每周称体重、测体温1次。给药前、给药1.5、3.0、4.5、6.0、7.5、9.0个月(停药次日)及恢复期结束(停药4周),分别检测心电图、眼科、免疫学、血液学、血液生化学、尿常规等指标。给药3.0、6.0、9.0个月及恢复期结束,每组随机解剖2只猴(9.0个月为4只),雌雄各半,观察脏器变化,计算脏器系数,并进行组织病理学观察。结果各剂量CTB-INS对恒河猴各指标检测结果均无明显影响,与对照组相似。各剂量组恒河猴脏器及其组织病理学检查均未发现异常,部分剂量组恒河猴的胸腺系数平均值高于对照组,肺脏系数、甲状腺系数平均值低于对照组。结论经口给予CTB-INS对恒河猴无明显毒副作用,未见中毒靶器官,无毒(影响)剂量为4.0 g/(kg·d)。

Objective To evaluate the safety of cholera toxin B subunit-insulin B chain fusion protein（CTB-INS） in rhesus. Methods Rhesus monkeys were randomly divided into four groups, ten for each, at equal numbers in both genders. The monkeys in high, moderate and low dose CTB-INS groups were orally administered with CTB-INS at dosages of 4. 0, 2. 0 and 1. 0 g /（kg·d） respectively, while those in control group with 10 ml /（kg·d） of 0. 5% sodium carboxymethyl cellulose（CMC-Na）, once a day at the same time points in morning for 9 months（6 times a week）. The monkeys were observed for clinical signs and recorded for food intake once a day, and measured for body temperature and weight once a week. The electrocardiogram, ophthalmology, as well as immunological, hematological and hemobiochemical indexes and result routine urine examination were observed before and 1. 5, 3. 0, 4. 5, 6. 0, 7. 5 and 9. 0（the second day after withdrawal） months after the first administration and at the end of recovery period（four weeks after the last administration）. Two monkeys in each group, one male and one female, were dissected 3. 0 and 6. 0 months, while four monkeys（two male and two female） 9. 0 months after the first administration and at the end of recovery period and observed for organs, based on which the organ coefficient was calculated, and the histopathology was observed. Results Various indexes of monkeys in high, moderate and low dose CTB-INS groups showed no significant difference with those in control group, while no abnormalities were observed in various organs or histopathological examination. However, the mean thymus coefficients in partial groups were higher, while the mean lung and thyroid coefficients were lower, than those in control group. Conclusion Oral administration with CTB-INS showed no obvious toxicity or adverse effect on rhesus, and no toxic target organs were observed. The non-toxic dose of CTB-INS was 4. 0 g /（kg·d）.