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研究力达霉素与抗Ⅳ型胶原酶单抗的不同连接键的小型化免疫偶联物的抗肿瘤作用。制备长短链的二硫键以及硫醚键抗肿瘤抗生素力达霉素(LDM)与抗Ⅳ型胶原酶单抗3G11的Fab’片段连接的免疫偶联物,ELISA法测定偶联物的免疫活性,体外细胞克隆形成测定和体内荷瘤裸小鼠模型观察两种偶联物的抗肿瘤作用。体外研究显示,Fab’-LDM偶联物部分保留了抗Ⅳ型胶原酶单抗3G11对抗原的亲和力,硫醚键偶联物对体外培养的人纤维肉瘤HT-1080细胞的杀伤作用强于LDM和二硫键偶联物。实验结果显示,LDM等剂量条件下硫醚键偶联物Fab’-SSMPB-LDM和Fab’-SMBS-LDM组的抑瘤率分别为87.7%和78.0%,二硫键偶联物Fab’-SPDP-LDM和Fab’-LCSPDP-LDM组分别为74.8%和72.3%,游离LDM组为70.9%,相比较长连接键的硫醚键偶联物抑瘤作用显著增强。Fab’-SSMPB-LDM和Fab’-SMBS-LDM组动物的平均生存时间延长为165.8%和145.2%,Fab’-SPDP-LDM和Fab’-LCSPDP-LDM组分别为82.2%和107.5%,LDM组为71.9%。与二硫键偶联物组比较,硫醚键偶联物组能显著延长荷瘤鼠的生存期;长硫醚键偶联物抗肿瘤作用和对荷瘤动物生存期延长作用强于短硫醚键偶联物。长链硫醚键偶联物Fab’-SSMPB-LDM肿瘤生长抑制作用提高,荷瘤动物生存期显著延长,有望开发为新的肿瘤治疗药物。
To study the antitumor effect of miniaturized immunoconjugates of different linkages between lewomycin and anti-type IV collagenase monoclonal antibodies. Preparation of immunoconjugates of long and short chain disulfide bonds and Fab ’fragments of thioether bond antitumor antibiotic lidamycin (LDM) and anti-type IV collagenase monoclonal antibody 3G11, and the conjugate was measured by ELISA , In vitro cell clonogenic assay and in vivo tumor-bearing nude mouse model were used to observe the antitumor effects of the two conjugates. In vitro studies showed that the Fab’-LDM conjugate partially retained the affinity of the anti-type IV collagenase monoclonal antibody 3G11 for the antigen and that the thioether conjugate had a greater cytotoxic effect on human fibrosarcoma HT-1080 cells in vitro than the LDM And disulfide bond conjugates. The experimental results showed that the antitumor rates of thioether bond-conjugated Fab’-SSMPB-LDM and Fab’-SMBS-LDM were 87.7% and 78.0%, respectively at the same dose of LDM. The disulfide bond Fab’- The antitumor activity of the thioether bond conjugate with SPDP-LDM and Fab’-LCSPDP-LDM was 74.8% and 72.3%, respectively, compared with 70.9% in the free LDM group. The mean survival time of Fab’-SSMPB-LDM and Fab’-SMBS-LDM animals was 165.8% and 145.2%, respectively, with 82.2% and 107.5% for Fab’-SPDP-LDM and Fab’-LCSPDP- Group was 71.9%. Compared with the disulfide conjugate group, the thioether conjugate group can significantly prolong the survival of tumor-bearing mice; the anti-tumor effect of the long-thioether conjugate and the effect of prolonging the survival of tumor-bearing animals are stronger than short sulfur Ether bond conjugate. Long-chain thioether bond Fab’-SSMPB-LDM tumor growth inhibition, tumor-bearing animals significantly prolonged survival is expected to be developed as a new cancer drug.