双靶向配体化力达霉素的急性和亚急性毒性

Acute and subacute toxicity of dual targeting ligand-based lidamycin

目的通过观察双靶向配体化力达霉素(DTLL)的急性和亚急性毒性,获得DTLL的半数致死剂量(LD_(50))、最大耐受剂量和相关生理病理等指标,为其安全性评价提供毒理学依据,为其临床应用研究奠定基础。方法①急性毒性试验:60只雌性各半的昆明小鼠按体质量随机分为正常对照组及DTLL 1.44,2.05,2.93,4.19和5.99 mg·m^(-2),24 h内经尾静脉单次注射给予DTLL,分别于第3,7和14天测定小鼠体质量,观察并记录小鼠给药后的毒性反应和死亡情况。②亚急性毒性试验:120只雌性各半的SD大鼠随机分为正常对照组及DTLL 61,122和245μg·m^(-2)组,尾静脉注射给予DTLL,每周给药1次,共给药4周,停药后恢复4周。每周检测大鼠体质量和摄食量。分别于第28和56天取血液、尿液和主要脏器组织,用Konelab PRIME 30全自动生化分析仪和尿液分析质控仪检测血液和尿液生化指标,同时用HE染色组织切片,观察其病理变化。结果①急性毒性试验:DTLL单次给药对昆明小鼠LD_(50)为2.45 mg·m^(-2),DTLL>2.05 mg·m^(-2)组小鼠体质量下降(P<0.05),出现行动迟缓和眼睑闭合等症状,并有小鼠陆续死亡。组织学检查显示,小鼠有不同程度的脾萎缩、髓外造血和肝、肾细胞坏死。②亚急性毒性试验:SD大鼠经尾静脉注射DTLL,28 d重复给药后计算所得最大耐受剂量为122μg·m^(-2),DTLL 61,122和245μg·m^(-2)组大鼠体质量降低(P<0.05)、摄食量减少(P<0.05);245μg·m^(-2)组停药后,谷丙转氨酶(P<0.01)和谷草转氨酶活性升高(P<0.05)。组织病理观察显示,245μg·m^(-2)组大鼠出现肝细胞病变、脾生发中心增大、肺炎症细胞浸润和肾小管嗜酸性病变等。结论DTLL可产生延迟性肝、肾毒性,并可能引起免疫抑制和造血功能障碍,其毒性作用的靶器官主要为肝、肾、脾和肺。

OBJECTIVE To investigate the acute and subacute toxicity of dual target ligand lidamycin(DTLL)and obtain the median lethal dose(LD_(50)),maximum tolerable dose(MTD)and related physiopathological indexes of DTLL in order to provide toxicological evidence for DTLL safety evaluation and contribute to clinical application of DTLL in the future.METHODS In acute toxicity tests,Kunming mice were randomly divided into the normal control group and a variety of groups treated with DTLL 1.44,2.05,2.93,4.19 and 5.99 mg·m^(-2),respectively.DTLL was injected via the mouse tail vein once within 24 h.Mouse body mass was measured on the 3^(rd),7^(th) and 14^(th) days after adminstration,followed by toxicity and lethal observation of mice and recording for additional 14 d.In subacute toxicity tests,SD rats were randomly divided into the normal control group and treatment groups at doses of DTLL 61,122 and 245μg·m^(-2),respectively.DTLL was administered by injection into the tail vein of rats,once a week for 4 weeks,before the rats were left recovering for 4 weeks after drug withdrawal.Body mass and food intake of SD rats were measured every week during experiments.On the 28^(th) and 56^(th) days,blood,urine and main organ samples were collected to detect a series of biochemical indexes in either blood or urinary samples by using a Konelab PRIME 30 automatic biochemical analyzer or an instrument for quality control of urinary samples,respectively.Finally,pathological changes were detected by HE staining.RESULTS Acute toxicity test:LD_(50) of DTLL at a single dose was 2.45 mg·m^(-2).DTLL at a dose above 2.05 mg·m^(-2) caused the loss of body mass(P<0.05),slow movement,eyelid closure,other symptoms,or even a succession of deaths.Additionally,in histological examination,spleen atrophy,extramedullary hematopoiesis and cellular necrosis in both the liver and kidney were observed.Subacute toxicity test:the maximum tolerated dose of DTLL was 122μg·m^(-2) after four continuous administrations during those 28 d.The body mass and food intake of SD rats in the groups treated with DTLL 61,122 and 245μg·m^(-2) were decreased(P<0.05).However,in the 245μg·m^(-2) group,the levels of glutamate pyruvate transaminase(GPT)(P<0.01)and glutamic oxaloacetic transaminase(GOT)(P<0.05)were significantly increased after drug withdrawal.In addition,the result from pathological examination showed hepatocyte lesion,enlargement in the splenic germinal center,infiltration of pulmonary inflammatory cells of lungs and renal tubular eosinophilic lesion in rats of the 245μg·m^(-2) group.CONCLUSION DTLL may cause delayed hepatotoxicity,nephrotoxicity as well as immunosuppression and hematopoietic dysfunction by mainly targeting the liver,kidney,spleen and lung.