Ranpirnase (Frog RNase) Targeted with a Humanized, Internalizing, Anti-Trop-2 Antibody Has Potent Cytotoxicity against Diverse Epithelial Cancer Cells
Ranpirnase (Frog RNase) Targeted with a Humanized, Internalizing, Anti-Trop-2 Antibody Has Potent Cytotoxicity against Diverse Epithelial Cancer Cells
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DOI：
10.1158/1535-7163.mct-10-0338
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发表时间：
2010-08-01
影响因子：
5.7
通讯作者：
Goldenberg, David M.
中科院分区：
文献类型：
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作者：
Chang, Chien-Hsing;Gupta, Pankaj;Goldenberg, David M.
Ranpirnase (Rap) 是一种两栖动物 RNase，作为抗肿瘤剂已在临床前和临床上进行了广泛研究。 Rap 可以对患者重复给药，不会产生任何不良免疫反应，据报道，可逆性肾毒性是剂量限制的。为了增强其效力和靶向肿瘤治疗，我们描述了一种新型基于 IgG 的免疫毒素的生成，命名为 2L-Rap(Q)-hRS7，其中包含 Rap (Q)（一种去除假定 N-糖基化位点的突变体 Rap）和 hRS7（一种针对 Trop-2（一种在多种上皮癌中过表达的细胞表面糖蛋白）的内化人源化抗体。该免疫毒素是通过将 Rap(Q) 与两条 hRS7 轻 (L) 链的 NH2 末端融合而重组产生的，在稳定转染的骨髓瘤细胞中产生，通过 Protein A 纯化，并通过一组体外研究进行评估。结果包括尺寸排阻高效液相色谱、SDS-PAGE、流式细胞术、RNase 活性、内化、细胞活力和集落形成，显示其纯度、分子完整性、与 hRS7 与不同癌症类型的几种表达 Trop-2 的细胞系结合的亲和力相当，以及在纳摩尔浓度下抑制这些细胞系生长的效力。此外，2L-Rap (Q)-hRS7 抑制了携带 Calu-3 人类非小细胞肺癌异种移植物的裸鼠预防性模型中的肿瘤生长，中位生存时间从 55 天延长至 96 天（P < 0.01）。这些结果保证了 2L-Rap(Q)-hRS7 的进一步开发，作为各种表达 Trop-2 的癌症的潜在治疗方法，例如宫颈癌、乳腺癌、结肠癌、胰腺癌、卵巢癌和前列腺癌。摩尔癌症治疗； 9(8)； 2276-86。 (C) 2010 AACR。
Ranpirnase (Rap), an amphibian RNase, has been extensively studied both preclinically and clinically as an antitumor agent. Rap can be administered repeatedly to patients without any untoward immune response, with reversible renal toxicity reported to be dose limiting. To enhance its potency and targeted tumor therapy, we describe the generation of a novel IgG-based immunotoxin, designated 2L-Rap(Q)-hRS7, comprising Rap (Q), a mutant Rap with the putative N-glycosylation site removed, and hRS7, an internalizing, humanized antibody against Trop-2, a cell surface glycoprotein overexpressed in variety of epithelial cancers. The immunotoxin was generated recombinantly by fusing Rap(Q) to each of the two hRS7 light (L) chains at the NH2 terminus, produced in stably transfected myeloma cells, purified by Protein A, and evaluated by a panel of in vitro studies. The results, including size-exclusion high-performance liquid chromatography, SDS-PAGE, flow cytometry, RNase activity, internalization, cell viability, and colony formation, showed its purity, molecular integrity, comparable affinity to hRS7 for binding to several Trop-2-expressing cell lines of different cancer types, and potency to inhibit growth of these cell lines at nanomolar concentrations. In addition, 2L-Rap (Q)-hRS7 suppressed tumor growth in a prophylactic model of nude mice bearing Calu-3 human non-small cell lung cancer xenografts, with an increase in the median survival time from 55 to 96 days (P < 0.01). These results warrant further development of 2L-Rap(Q)-hRS7 as a potential therapeutic for various Trop-2-expressing cancers, such as cervical, breast, colon, pancreatic, ovarian, and prostate cancers. Mol Cancer Ther; 9(8); 2276-86. (C) 2010 AACR.