Study On The Toxicity Of Recombinant Anti VEGF Humanized Monoclonal Antibody Injection In Cynomolgus Monkeys

PART ONE 4-WEEK TOXICITY EXPLORATION ANDRESEARCH OF RECOMBINANT ANTI-VEGFHUMANIZED MONOCLONALANTIBODYObjective: The purpose of this study was to evaluatethetoxicity of the test article,anti-VEGF humanized monoclonal antibody,when administered twice weekly via intravenous injection to cynolmogus monkeys for 4 weeks,and to assess the reversibility after a 4-week recovery phase.Methods: 30 health monkeys were randomly assigned into 5 groups based on the weights: vehicle contro,low-SMMU-13,midSMMU-13,high-SMMU-13 dose and positive control groups.6 animals in each group(3 males and 3 females).The doses for the low-SMMU-13,mid-SMMU-13,and high-SMMU-13 dose groups were 2.0,10.0 and 50.0 mg/kg;the dose for the positive control animals was 50.0 mg/kg;and the vehicle control animals were administered the placebo.The dose volume was 2 m L/kg.Results:(1),Clinical chemistry: the LDH in the animals of the mid-SMMU-13,high-SMMU-13 dose and positive control groups increased.No other test article or positive article related findings were noted.(2),Microscopic Examinations: the dose-dependent femoral epiphyseal plate chondrocytes hyperplasia and calcified osteogenesis in one male in positive control group and 2 males in high-SMMU-13 dose group were found.These findings were noted on 1 male in high-dose group and 1 male in positive control group.This finding was not noted in the male animals.Inflammatory cell infiltration in the proximal vascular wall,blood vessels around the bleeding and inflammatory cells,a small number of blood vessels around the visible proliferation of fibrous connective tissue or granulation tissue in all of the animals in the vehicle control group and several animals in other groups.In the mid-SMMU-13 and high-SMMU-13 dose groups,theendothelial cells in the inner wall of the animal's internal wall werenecrotic,necrotic or local necrosis of the wall,bleeding of the celluloseand local fibrosis.Those findings were recoverd in the recovery phase,only several animals have inflammatory cells in the vessel walls.Therewere still bleeding around the vessel.Conclusion: The dose range for thisstudy was 2 mg/kg to 50 mg/kg,the safe dose for the cynolmogusmonkeys when administered the anti-VEGF humanized antibody is 2mg/kg,the toxicity dose is 10 mg/kg.The main toxic effect is thefemoral epiphyseal plate chondrocytes hyperplasia,calcificationosteogenesis,the damage in the site of proximal blood vesselsinflammation.The target organ is the femoral epiphysis and the injectionsites.These findings are the extension of the pharmacological effects ofthe test article.The test article could affect the digestive system(liver).In this study,the test article has the similar toixic effect andcharacterization with the positive control article.Part Two 4-week Toxicokinetics Exploration andResearch of Recombinant anti-VEGF humanizedMonoclonal antibodyObjective: The purpose of this study was to determine and compare the toxicokinetics of the test article,recombinant anti-VEGF humanized Monoclonal antibody,with reference medicinal product(RMP),Avastin,when administered for 4 weeks with a 4-week recovery phase.The blood concentration-time change(TK)were determined before and after dose on Day 1,11,18 and 25 of the dosing phase,and Day 32,39,46 and 57 of the recovery phase.To provided more data for the toxicity study.Mehotds:24 healthy animals were randomly assigned into 4 groups based on the body weights: low-SMMU-13,mid-SMMU-13,and high-SMMU-13 dose groups and the positive control group.6 animals in each group(3 males and 3 females).The low-SMMU-13,mid-SMMU-13,and highSMMU-13 dose groups were administered 2.0,10.0 and 50.0 test article.The positive control group were administered 50 mg/kg Avastin.The test article and the positive article will be adminstered twice weekly to the animals for 4 weeks.Determined the concentration of the test article or positive article in the serum before or after dosing on Day 1,11,18 and 28 of the dosing phase,and Day 32,39,46,and 57 of the recovery phase.And the TK parameters were also calculated.The concentration was tested with the ELISA method.Results: The Cmax of the low-SMMU-13,midSMMU-13,and high-SMMU-13 dose groups on Day 25 is 150%,110%,and 140% higher than Day 1;and the AUC(0-t)is 220%,120%,and 210% higher than Day 1.The accumulation rate in low-SMMU-13,midSMMU-13,and high-SMMU-13 dose groups were 3.3±0.5?2.4±1.6 and 3.1±0.6.The Cmax and AUC(0-t)on Day 25 were 20% and 40% higher than Day 1 after adminsteration of the Avastin.The accumulation rate is 1.3±1.1.Conclusion: Under this experimental condition,the test article were accumulated in the animals,which is related to the long t1/2 of the test article and the procedure to achieve the steady status.Both the test article and the positive control article demonstrated similar toxicokinetic profile in cynomolgus monkeys.Part Three 4-week Safety Pharmacology Study ofRecombinant anti-VEGF humanized MonoclonalantibodyObjective: This study was conducted to evaluate the effects before or after the administration of the recombinant anti-VEGF humanized monoclonal antibody on the cardiovascular system and respiratory system of cynomolgus monkeys.Methods: 30 healthy cynolmogus monkeys were randomly assigned into 5 groups based on the body weights: vehicle control group,low-SMMU-13,mid-SMMU-13,and highSMMU-13 dose group,and positive control group.6 animals in each group: 3 males and 3 females.The low-SMMU-13.Mid-SMMU-13,and highSMMU-13 dose groups were administered 2.0,10.0,and 50.0 mg/kg test article.The positive control group was administered 50.0 mg/kg Avastin.The vehicle control group was administered the placebo.The dose volume is 2 ml/kg.Monitoring the blood pressure,ECGs changes before and 2h,8h,24 h and 48 h post dosing.Results: The single dose of the recombinant anti-VEGF humanized monoclonal antibody had no significant effect on the systolic blood pressure(SBP),diastolic blood pressure(DBP),mean arterial pressure(MAP),and the ECGs parameter: heart rate,respiration,P wave,R wave,T wave,QRS duration,PR interval,QT interval and ST interval.Conclusion: here were no test article(2.0~50.0 mg/kg)or positive control article(50.0 mg/kg)changes on the cardiovascular system and respiratory system of the cynomolgus monkeys after the single dose.