Therapeutic Monoclonal Antibodies Inhibiting Proteases of Biomedical Importance

项目总结 作为重要的信号分子，蛋白水解酶精确地控制着广泛的生理过程 在健康和疾病方面，因此是最大的药物靶标家族之一。尽管几十年来 在密集的努力下，传统的药物发现战略仅通过瞄准 所有与治疗相关的蛋白酶中的一小部分。这是因为小分子抑制剂往往缺乏 有效和安全的基于蛋白酶的治疗所需的特异性和/或适当的药代动力学特性。 在这些方面，单抗(MAbbs)正在成为有吸引力的替代品，具有重要的意义 具有选择性高、血清半衰期长、有可能跨越血脑屏障和AS等优点。 可诱导的前药。自从杂交瘤技术发明以来，单抗的研究取得了巨大的进展。 发现和工程。然而，常规发现的蛋白酶抑制单抗仍然是一个相当大的 一般情况下，由于人抗体副表位对酶抑制不相容，以及缺乏 功能性高通量筛选方法。我的实验室一直致力于开发 简化方法，促进产生作为安全和有效的蛋白酶的治疗性单抗 抑制剂。在过去的五年里，我们取得了重大进展，建立了一系列小说 技术，包括骆驼启发的凸状副表位人类抗体库和基于抑制的 而不是基于绑定的选择/筛选方法。结合这些可行的方法，我们发现， 鉴定和优化抑制生物医学中多种蛋白水解酶的有效和特定的单抗组合 重要性。此外，我们的蛋白酶抑制单抗在小鼠身上显示了显著的治疗效果。 癌症、神经性疼痛、肥胖和中风的模型。通过克服长期存在的挑战，这些 成就开启了激动人心的机遇。在未来五年，我们将扩大我们的强大 技术对许多其他有充分证据的蛋白酶来说，其中迫切需要治疗性抑制物。 此外，我们将开发更多的技术，以实现独特的和治疗所需的特征：(1) 功能特异性(底物依赖)抑制，(2)对一组蛋白酶的广谱抑制，以及(3) 通过合理的设计进行表位特异性抑制。总体而言，据估计，蛋白酶占5%-10% 在所有药物靶标中，已为药物开发研究过。拟议研究的完成将 毫不含糊地推进针对生物医学重要蛋白酶的治疗性mAb的开发，例如针对 目前的危险，SARS-CoV-2，通过抑制TMPRSS2(II型跨膜丝氨酸蛋白酶)作为广泛的 中和方法，没有不需要的抗体依赖增强。

PROJECT SUMMARY As important signaling molecules, proteases precisely control a wide variety of physiological processes both in health and in diseases, and thus represent one of the largest families of pharmaceutical targets. Despite decades of intensive efforts, conventional drug discovery strategies have only achieved a limited success by targeting a small fraction of all therapeutically relevant proteases. It is because small-molecule inhibitors are often lack of specificity and/or appropriate pharmacokinetic properties required for effective and safe protease-based therapy. In these aspects, monoclonal antibodies (mAbs) are emerging as attractive alternatives with significant advantages such as high selectivity, long serum half-life, potential to cross the blood-brain barrier, and as inducible prodrugs. Since the invention of hybridoma technology, tremendous progress has been made in mAb discovery and engineering. However, routine discovery of protease-inhibiting mAbs is still a considerable challenge in general, due to the incompatibility of human antibody paratope for enzyme inhibition, and lack of functional high-throughput screening methods. My laboratory has been committed to the development of streamlined methodologies that facilitate the generation of therapeutic mAbs as safe and effective protease inhibitors. Over the past five years, we have made significant progress, and established a series of novel technologies, including camelid-inspired convex paratope human antibody libraries, and inhibition-based rather than binding-based selection/screening methods. Combining these enabling approaches, we discovered, characterized, and optimized panels of potent and specific mAbs inhibiting numerous proteases of biomedical importance. Furthermore, our protease inhibitory mAbs have shown significant therapeutic efficacy in mouse models of cancers, neuropathic pains, obesity, and stroke. By overcoming longstanding challenges, these achievements have opened the exciting opportunity. In the next five years, we will extend our powerful technologies to many other well-documented proteases, of which therapeutic inhibitors are urgently needed. Furthermore, we will develop additional technologies to achieve unique and therapy-desirable features: (1) function-specific (substrate-dependent) inhibition, (2) broad-spectrum inhibition on a group of proteases, and (3) epitope-specific inhibition by rational design. Overall, it has been estimated that proteases account for 5-10% of all drug targets have been studied for pharmaceutical development. The completion of proposed research will unambiguously advance therapeutic mAb developments targeting biomedically important proteases, e.g. against the present danger, SARS-CoV-2, by inhibiting TMPRSS2 (type II transmembrane serine protease) as a broad neutralization approach without the unwanted antibody-dependent enhancement.