抗HER2靶点抗体偶联药物食蟹猴重复给药毒性研究

Repeated dose toxicity of humanized anti-HER2 antibody-drug conjugate in cynomolgus monkeys

目的:通过对食蟹猴静脉重复给药注射用重组抗HER2人源化单克隆抗体偶联美登素衍生物DM1,对其进行临床前安全性评价,为临床不良反应监测和设计人用剂量提供参考依据。方法:食蟹猴随机分成4组,包括空白对照组、低(3 mg·kg-1)、中(10 mg·kg-1)、高(30 mg·kg-1)剂量组,每组10只动物,雌雄各半。静脉注射给药,每3周给药1次,连续给药4次,给药周期9周,恢复期6周。分别在检疫期、首次给药后、给药结束、恢复期结束的不同时间点进行动物临床症状、体重、摄食量、体温、心电图、血压、尿分析、血液学、血清生化及组织病理学检查等各项指标检测。结果:给药后,动物出现一定程度不良反应。高剂量会引起动物不耐受,1只雌性动物在给药d 63濒死处死。毒性症状包括脱毛、消瘦、四肢无力、活动减少、稀便、注射部位刺激性。会引起动物体重下降、摄食减少、体温、血压下降。血液学检查,动物白系细胞WBC,Neut,Mono升高;红系细胞Retic,APTT升高,RBC,HGB,HCT,MCHC下降。血清生化检查,动物血清ALT,AST,TP,ALP,CK,LDH,TG,Ig A,IgG,IgM升高; CRE,GLU,ALB,Cl-,A/G下降。停药恢复期,上述多数改变均可恢复。组织病理学检查,发现胸腺体积减小,肝脏、肾脏、脾脏、胸腺、舌、皮肤、肾上腺细胞有丝分裂项增加,脊髓背索轴索、坐骨神经髓鞘轴索变性。结论:食蟹猴反复给予注射用重组抗HER2人源化单克隆抗体偶联美登素衍生物DM1后,低、中剂量下动物有良好的耐受性,作用毒性靶器官为肝脏、脾脏、胸腺、脊髓、坐骨神经,实验中未见毒性反应剂量(NOAEL)≤3 mg·kg-1。上述结果为药物开展临床试验提供了数据支持。

Objective:To conduct a repeat-dose toxicity study of humanized anti-HER2 antibody and drug DM1 conjugate for injection in cynomolgus monkeys to evaluate the preclinical safety and supply the evidence for adverse effects monitor and clinical dosing design.Methods:Cynomolgus monkeys were randomly divided into four groups including vehicle control,low-,middle-,and high-dose of humanized anti-HER2 antibody-drug conjugate groups(3,10 and 30 mg·kg-1).Each group contained ten monkeys with female and male in half.The animals were intravenously administered once every three weeks constantly four times for nine weeks following a 6-weekrecovery phase.A series of toxicological parameters including clinical signs,body weight,food consumption,body temperature,electrocardiogram,blood pressure,urinalysis,hematology,serum biochemistry and histopathological examination were detected at the quarantine period,after the first dosing,the end of dosing and recovery period.Results:After dosing,the animals developed obvious toxic reactions related to the test article.One female animal in the high-dose group appearing intolerance state was found moribund at day 63.The other notable adverse effects included shed,maransis,myasthenia,diarrhea,activity decrease and injection site irritation.Test article also led body weight,food consumption,temperature and blood pressure.Compared with the control,WBC,Neut,Mono,Retic and APTT levels were elevated,while RBC,HGB,HCT,and MCHC were descended in the animals of treated groups in the hematological examination.For the serum biochemical analysis,the test article could lead ALT,AST,TP,ALP,CK,LDH,TG,IgA,IgG and IgM levels,and decreased CRE,GLU,ALB,Cl-,and A/G levels.Almost changes mentioned above could be reversed during the recovery phase.In the histopathological examinations,thymus shrunk and cellular mitotic arrest were found spread widely in liver,kidney,spleen,thymus,tongue,skin and adrenal glands.Axonal degeneration in the sciatic nerve and dorsal funiculus of the spinal cord were also observed.Conclusion:According to our research,low and middle dose of humanized antiHER2 antibody and drug DM1 conjugate for injection were well tolerated in cynomolgus monkeys,while the high dose was not tolerable.The toxic target organs were liver,spleen,thymus,spinal cord and sciatic nerve.The NOAEL is not more than 3 mg·kg-1.These data has provided reference for monitoring the clinical adverse effects of humanized anti-HER2 antibody-drug conjugate for injection.