Wu Peidong, Prachyathipsakul Theeraphop, Huynh Uyen, Qiu Jingyi, Jerry D Joseph, Thayumanavan S
Department of Chemistry, University of Massachusetts, Amherst, Massachusetts 01003, United States.
Department of Biomedical Engineering, University of Massachusetts, Amherst, Massachusetts 01003, United States.
Bioconjug Chem. 2023 Mar 27. doi: 10.1021/acs.bioconjchem.3c00034.
Targeted delivery of therapeutics using antibody-nanogel conjugates (ANCs) with a high drug-to-antibody ratio has the potential to overcome some of the inherent limitations of antibody-drug conjugates (ADCs). ANC platforms with simple preparation methods and precise tunability to evaluate structure-activity relationships will greatly contribute to translating this promise into clinical reality. In this work, using trastuzumab as a model antibody, we demonstrate a block copolymer-based ANC platform that allows highly efficient antibody conjugation and formulation. In addition to showcasing the advantages of using an inverse electron-demand Diels-Alder (iEDDA)-based antibody conjugation, we evaluate the influence of antibody surface density and conjugation site on the nanogels upon the targeting capability of ANCs. We show that compared to traditional strain-promoted alkyne-azide cycloadditions, the preparation of ANCs using iEDDA provides significantly higher efficiency, which results in a shortened reaction time, simplified purification process, and enhanced targeting toward cancer cells. We also find that a site-specific disulfide-rebridging method in antibodies offers similar targeting abilities as the more indiscriminate lysine-based conjugation method. The more efficient bioconjugation using iEDDA allows us to optimize the avidity by fine-tuning the surface density of antibodies on the nanogel. Finally, with trastuzumab-mertansine (DM1) antibody-drug combination, our ANC demonstrates superior activities in vitro compared to the corresponding ADC, further highlighting the potential of ANCs in future clinical translation.
使用具有高药物与抗体比率的抗体-纳米凝胶偶联物（ANCs）进行治疗药物的靶向递送，有可能克服抗体-药物偶联物（ADCs）的一些固有局限性。具有简单制备方法和精确可调性以评估构效关系的ANC平台，将极大地有助于将这一前景转化为临床现实。在这项工作中，我们以曲妥珠单抗为模型抗体，展示了一种基于嵌段共聚物的ANC平台，该平台允许高效的抗体偶联和制剂制备。除了展示使用基于逆电子需求狄尔斯-阿尔德（iEDDA）的抗体偶联的优势外，我们还评估了抗体表面密度和偶联位点对纳米凝胶的影响，以及它们对ANCs靶向能力的影响。我们表明，与传统的应变促进炔基-叠氮环加成反应相比，使用iEDDA制备ANCs的效率显著更高，这导致反应时间缩短、纯化过程简化，并增强了对癌细胞的靶向性。我们还发现，抗体中的位点特异性二硫键重新桥连方法与更随意的基于赖氨酸的偶联方法具有相似的靶向能力。使用iEDDA进行的更高效生物偶联使我们能够通过微调纳米凝胶上抗体的表面密度来优化亲和力。最后，在曲妥珠单抗-美登素（DM1）抗体-药物组合中，我们的ANC在体外表现出比相应的ADC更优异的活性，进一步突出了ANCs在未来临床转化中的潜力。
