Synthesis of the next-generation therapeutic antibodies that combine cell targeting and antibody-catalyzed prodrug activation
Synthesis of the next-generation therapeutic antibodies that combine cell targeting and antibody-catalyzed prodrug activation
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DOI：
10.1073/pnas.0700223104
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发表时间：
2007-03-27
影响因子：
11.1
通讯作者：
Sinha, Subhash C.
中科院分区：
文献类型：
--
作者：
Abraham, Sunny;Guo, Fang;Sinha, Subhash C.
在癌症治疗中，利用催化抗体进行选择性前药激活的一个障碍是全身肿瘤靶向。在这里，我们报告了催化性抗体的产生，它有效地靶向肿瘤细胞，并保持前药激活能力。AB偶联物由整合素α(V)β(3)靶向拮抗剂通过铰链区二硫键还原生成的半胱氨酸残基的硫基或不参与催化活性的表面赖氨酸残基与催化抗体38C2共价偶联而成。利用流式细胞术，抗体结合物被证明能有效地与表达整合素α(V)β(3)的人乳腺癌细胞结合。抗体结合物还保留了其亲本催化抗体38C2的逆转-羟醛脱氢酶的活性，通过甲基碘醇和阿霉素(DOX)前药激活来衡量。作为对这些抗体结合物的补充，设计并合成了一套进化的DOX前药。在存在和不存在整合素α(V)β(3)靶向38C2结合物的情况下，使用人乳腺癌细胞对表现出较高稳定性和较低毒性的DOX前药进行了评估。我们的研究表明，细胞靶向和前药激活能力可以有效地结合用于新的DOX前药的选择性化疗。
An obstacle in the utilization of catalytic Abs for selective prodrug activation in cancer therapy has been systemic tumor targeting. Here we report the generation of catalytic Abs that effectively target tumor cells with undiminished prodrug activation capability. Ab conjugates were prepared by covalent conjugation of an integrin alpha(v)beta(3)-targeting antagonist to catalytic Ab 38C2 through either sulfide groups of cysteine residues generated by reduction of the disulfide bridges in the hinge region or surface lysine residues not involved in the catalytic activity. Using flow cytometry, the Ab conjugates were shown to bind efficiently to integrin alpha(v)beta(3)-expressing human breast cancer cells. The Ab conjugates also retained the retro-aldol activity of their parental catalytic Ab 38C2, as measured by methodol and doxorubicin (dox) prodrug activation. Complementing these Ab conjugates, an evolved set of dox prodrugs was designed and synthesized. Dox prodrugs that showed higher stability and lower toxicity were evaluated both in the presence and absence of the integrin alpha(v)beta(3)-targeting 38C2 conjugates for cell-killing efficacy by using human breast cancer cells. Our study reveals that cell targeting and prodrug activation capabilities can be efficiently combined for selective chemotherapy with novel dox prodrugs.