平阳霉素与单克隆抗体3D6及其Fab′片段偶联物对肝癌的实验治疗  

Antitumor effects of the conjugates of pingyangmycin linked to monoclonal antibody 3D6 and its Fab′ fragment on hepatoma in mice

作　　者：刘秀均[1] 江敏[1] 刘小云[1] 甄永苏[1] 

机构地区：[1]中国医学科学院中国协和医科大学医药生物技术研究所,北京100050

出　　处：《中国抗生素杂志》2002年第8期496-501,共6页Chinese Journal of Antibiotics

基　　金：国家自然科学基金重点项目资助 (批准号 :39930 1 90 )

摘　　要：单抗 3D6是抗IV型胶原酶单克隆抗体。以葡聚糖T 40 (DextranT 40 )为中介体制备 3D6和平阳霉素 (PYM )的偶联物 3D6 PYM。采用间接ELISA法、TTC抑菌效价检测法、克隆形成法、MTT法及动物移植性肿瘤模型测定偶联物的生物学活性与体内外抗肿瘤作用。实验表明 ,3D6 PYM偶联物与PG、KB、C2 6及H2 2细胞等呈免疫学阳性反应 ;3D6 PYM偶联物对BEL 740 2、KB和H2 2细胞的细胞毒作用强于游离PYM。应用小鼠移植性肝癌模型观察偶联物对肝癌生长的抑制作用及长期疗效。结果显示 ,3D6 PYM偶联物对肿瘤生长有较强的抑制作用且在体内作用时间延长 ,发挥较好的疗效。第 2 1天时 ,按 10mg/kg剂量比较 ,3D6 PYM偶联物、3D6与PYM混合物及PYM的抑瘤率分别达到 99.7%、95 .6%、81.5 %。停止治疗后 ,继续观察各组动物的生存时间 ,对照组的中位生存期为 3 8d ,按 10mg/kg剂量比较 ,3D6 PYM偶联物组、3D6与PYM混合物组以及PYM组的中位生存期分别为 >12 0、88和 5 4d。 12 0d时 ,3D6 PYM偶联物 10mg/kg组未见动物死亡。以 β 环糊精为中介体制备单抗 3D6Fab′片段与PYM偶联物Fab′ PYM ,小鼠移植性肝癌模型皮下接种后第 7天给药 ,Fab′ PYM偶联物组与PYM抑瘤率分别为 81.0 %和 2 9.1%。以上研究结果表明 ,3D6 PYM及Fab′ PYM偶联?D6, a monoantibody directed against type IV collagenase and its Fab′ fragment were conjugated respectively to pingyangmycin (PYM), an antitumor antibiotic, via the intermediate agent dextran T 40 and β cyclodextrin to produce 3D6 PYM and Fab′ PYM conjugates. By ELISA, 3D6 PYM showed immunoreactivity with type IV collagenase and antigen related cancer cells, such as KB, PG, C26 and H22 cells. Determined by clonogenic assay and MTT assay the 3D6 PYM conjugate showed more potent cytotoxicity to BEL 7402, KB and H22 cells than free PYM. Hepatoma 22 (H22) transplanted sc in mice and the animals were treated by iv route for a total of 8 injections. At dosage of 10 mg/kg, 3D6 PYM conjugate, 3D6 and PYM mixture, and free PYM inhibited the growth of H22 tumor by 99.7%, 95.6% and 81.5% ( P <0.01) on the 21th day, respectively. The inhibitory effects of 3D6 PYM conjugate were much higher than that of 3D6 plus PYM mixture and that of free PYM ( P <0.01). 3D6 PYM markedly increased the survival time of the tumor bearing mice. Antitumor effect of Fab′ PYM conjugate was also evaluated in mice bearing H22. Fab′ PYM conjugate suppressed the growth of H22 by much higher inhibitory rates than free PYM. These studies suggest that the immunoconjugates consisting of PYM and monoclonal antibody or its Fab′ fragment are more effective than free PYM against hepatoma in mice.

关 键 词：平阳霉素 单克隆抗体 实验治疗 免疫偶联物 肝癌 动物肿瘤模型 

分 类 号：R73－361[医药卫生—肿瘤] R965.1[医药卫生—临床医学]