Comparison of recombinant and synthetically formed monoclonal antibody-β-lactamase conjugates for anticancer prodrug activation
Comparison of recombinant and synthetically formed monoclonal antibody-β-lactamase conjugates for anticancer prodrug activation
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DOI：
10.1021/bc990075w
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发表时间：
1999-11-01
影响因子：
4.7
通讯作者：
Senter, PD
中科院分区：
文献类型：
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作者：
Kerr, DE;Vrudhula, VM;Senter, PD
L49 单克隆抗体的缀合物（与黑色素瘤和癌上的 p97 抗原结合）是通过使用异双功能交联试剂将阴沟肠杆菌 β-内酰胺酶 (bL) 连接到 L49-Fab' 片段或使用 DNA 重组技术将该酶连接到 L49-sFv 来形成的。由此形成的缀合物L4g-Fab'-bL和L49-sFv-bL被设计用于在抗原阳性肿瘤细胞表面激活含有抗癌前药的头孢菌素。使用两种肺癌细胞系的体外实验结果表明，缀合物在影响头孢菌素氮芥前药 CCM 释放苯二胺芥方面具有同等活性。虽然用任一缀合物结合最大耐受剂量的 CCM 治疗可以治愈裸鼠中已建立的 SN12P 肾细胞癌肿瘤，但只有 L49-sFv-bL 缀合物在 CCM 最大耐受剂量的 1/4 时仍保持其这样做的能力。 L49-sFv-bL 在 1934J 肾细胞癌肿瘤模型中也优于 L49-Fab'-bL，并且在两种人肺癌体内模型中表现出相当活跃。这些结果表明，重组融合蛋白比化学缀合物具有更显着的治疗窗，并且在一系列人类肿瘤模型中具有活性。
Conjugates of the L49 monoclonal antibody (binds to the p97 antigen on melanomas and carcinomas) were formed by attaching Enterobacter cloacae beta-lactamase (bL) to the L49-Fab' fragment using a heterobifunctional cross-linking reagent or by linking the enzyme to L49-sFv using DNA recombinant technology. The conjugates thus formed, L4g-Fab'-bL and L49-sFv-bL, were designed to activate cephalosporin containing anticancer prodrugs at the surfaces of antigen positive tumor cells. Results from in vitro experiments using two lung carcinoma cell lines demonstrated that the conjugates were equally active in effecting the release of phenylenediamine mustard from the cephalosporin nitrogen mustard prodrug CCM. While treatment with either of the conjugates combined with the maximum tolerated doses of CCM led to cures of established SN12P renal cell carcinoma tumors in nude mice, only the L49-sFv-bL conjugate maintained its ability to do so at 1/4 the maximum tolerated dose of CCM. L49-sFv-bL was also superior to L49-Fab'-bL in the 1934J renal cell carcinoma tumor model and was shown to be quite active in two in vivo models of human lung carcinoma. These results demonstrate that the recombinant fusion protein leads to more pronounced therapeutic windows than the chemical conjugate and is active in an array of human tumor models.