Site-specific antibody-toxin conjugates for cancer therapy

用于癌症治疗的位点特异性抗体-毒素缀合物

• 批准号: 8909974
• 负责人: Joseph J Bellucci
• 金额: $ 3.38万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8909974
• 关键词: Adverse effects; Affinity; Antibodies; Antigen Targeting; Antineoplastic Agents; Bacterial Proteins; Binding; Biodistribution; Cells; Development; Diphtheria Toxin; Drug Kinetics; Enzymes; Eukaryotic Cell; Evaluation; Fusion Toxin; Genetic; Goals; Half-Life; Health; Human; Immunoglobulin Fragments; Immunotoxins; In Vitro; Inclusion Bodies; Lead; Length; Ligation; Link; Location; Mediating; Methodology; Methods; Modality; Monoclonal Antibodies; Mus; Mutate; Parents; Peptides; Pharmaceutical Preparations; Plasma; Prokaryotic Cells; Property; Protein Biosynthesis; Proteins; Pseudomonas aeruginosa toxA protein; Reaction; Receptor Protein-Tyrosine Kinases; Recombinant Fusion Proteins; Site; Solid Neoplasm; System; Targeted Toxins; Technology; Therapeutic; Tissues; Toxin; Toxin Conjugates; Tumor Antigens; anti-cancer therapeutic; base; cancer therapy; cell killing; cell transformation; cytokine; cytotoxicity; immunogenicity; in vivo; killings; next generation; pre-clinical; protein expression; sortase; stoichiometry; tool; treatment strategy; tumor; tumor xenograft

DESCRIPTION (provided by applicant): Antibodies offer a means to harness the extreme potency of bacterial protein toxins-capable of killing both healthy and transformed cells at picomolar concentrations-though no methods are currently available to link these toxins to high-affinity, bivalent mAbs. Site-specific attachment is critical in this application, as both proteins contain regions that cannot be perturbed in order to maintain tumor targeting and cytotoxicity. In principle, this could be accomplished by producing a mAb-toxin as a recombinant fusion protein. However, this is not possible with current technology, as the eukaryotic cells required to produce mAbs are killed by expression of protein toxins. We have characterized two reactions catalyzed by the bacterial enzyme sortase A (SrtA) that allow the mAb and the toxin to be produced separately by effective, proven methods, then site- specifically joined in vitro. We believe that this will provide a general mechanism to conjugate protein toxins such as diphtheria toxin and pseudomonas exotoxin A to mAbs independent of their target antigen, providing a means to generate antibody-toxin conjugates against a variety of tumor antigens for which mAbs are already available.

描述（由申请人提供）：抗体提供了一种利用细菌蛋白毒素的极端效力的方法-能够在皮摩尔浓度下杀死健康和转化细胞-尽管目前没有方法将这些毒素与高亲和力、二价mAb连接。位点特异性连接在该应用中是至关重要的，因为两种蛋白质都含有不能被扰动的区域，以保持肿瘤靶向和细胞毒性。原则上，这可以通过产生作为重组融合蛋白的mAb-毒素来实现。然而，这在当前技术下是不可能的，因为产生mAb所需的真核细胞被蛋白毒素的表达杀死。我们已经表征了由细菌酶分选酶A（SrtA）催化的两个反应，其允许通过有效的、经证实的方法分别产生mAb和毒素，然后在体外位点特异性地连接。我们相信，这将提供一种将蛋白质毒素如白喉毒素和假单胞菌外毒素A偶联到不依赖于其靶抗原的mAb上的一般机制，提供一种产生针对多种肿瘤抗原的抗体-毒素偶联物的方法，对于这些肿瘤抗原，mAb已经是可用的。