Novel Reagents for Rapid and Stable Thiol-Based Bioconjugations

用于快速稳定的硫醇基生物共轭的新型试剂

• 批准号: 10408820
• 负责人: Brian Matthew Zeglis
• 金额: $ 32.93万
• 依托单位: HUNTER COLLEGE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-09 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10408820
• 关键词: Address; Antibodies; Biodistribution; Breast Cancer Model; Cancer Model; Chelating Agents; Chemistry; Clinic; Colorectal Cancer; Deferoxamine; Development; Dimethyl sulfone; Dissociation; ERBB2 gene; Exhibits; Extravasation; Generations; Goals; Gold; Health; Image; Immunoconjugates; Immunoglobulins; Isotopes; Kinetics; Label; Laboratories; Lead; Libraries; Ligation; Maleimides; Medicine; Methods; Modeling; Modification; Neuroblastoma; Patient-Focused Outcomes; Patients; Pentetic Acid; Performance; Physiological; Positron-Emission Tomography; Prosthesis; Protocols documentation; Radiation Dose Unit; Radioactive; Radioimmunoconjugate; Radioimmunotherapy; Radioisotopes; Radiolabeled; Reaction; Reagent; Secure; Site; Solubility; Sulfhydryl Compounds; Technology; Testing; Therapeutic; Tissues; Trastuzumab; Treatment Protocols; Variant; Work; analog; base; clinical care; clinical efficacy; comparative; contrast imaging; design; experimental study; immunoreactivity; improved; in vivo; in vivo evaluation; member; mouse model; novel; nuclear imaging; pre-clinical; rational design; thioether; tool; uptake

Project Summary/Abstract For over three decades, the ligation between maleimide-bearing probes and thiols has been a cornerstone of the synthesis of site-specifically labeled antibodies. Yet despite its popularity, this approach to bioconjugation has serious limitations. Maleimide-based conjugates display limited stability in vivo because their succinimidyl thioether linkage can undergo a retro-Michael reaction that leads to the dissociation of the cargo or its exchange with circulating biomolecules. In the context of nuclear imaging and radioimmunotherapy, this retro- Michael reaction can lead to the release of the radioactive payload and the in vivo radiolabeling of endogeneous biomolecules, resulting in higher radiation doses to healthy tissues, reduced imaging contrast, and lower therapeutic ratios. In order to circumvent this obstacle, we have developed a modular, stable, and easily accessible phenyloxadiazolyl methyl sulfone reagent — `PODS' — as a platform for thiol-based bioconjugations. We have demonstrated that PODS can be used to reproducibly create homogenous, well- defined, highly immunoreactive, and highly stable radioimmunoconjugates with far superior in vivo performance compared to analogous probes synthesized via traditional, maleimide-based approaches. This proposal is centered upon the expansion and optimization of this technology. We will seek to build upon our preliminary work by optimizing the rapidity and selectivity of the PODS-thiol ligation and evaluating the in vivo performance of PODS-based radioimmunoconjugates labeled with 89Zr, 177Lu, 131I, and 225Ac. Specific Aim 1 (SA1) will be focused on the design, synthesis, and characterization of a library of second- generation PODS reagents, with the overall goal of identifying a single construct with the most favorable combination of stability, solubility, and reactivity. Specific Aim 2 (SA2) will be centered on the evaluation of the in vivo performance of radioimmunoconjugates synthesized using PODS. The most promising second- generation PODS reagent from SA1 will be used to synthesize 89Zr- and 177Lu-labeled radioimmunoconjugates, and their in vivo performance will be assessed in mouse models of cancer and compared to that of radiolabeled antibodies created using maleimide-based bioconjugations. Specific Aim 3 (SA3) will focus on the development of PODS-based prosthetic groups for the site-specific radiolabeling of antibodies with 131I and 225Ac. The in vivo performance of 131I- and 225Ac-labeled antibodies synthesized using these prosthetic groups will be evaluated in murine models of cancer and compared to analogues synthesized using current `gold- standard' strategies. We believe that this proposal could have a significant near-term impact on clinical care by developing and validating tools for the synthesis of highly stable radioimmunoconjugates with excellent in vivo performance, thereby improving imaging protocols, treatment regimens, and patient outcomes. Furthermore, we contend that this work could have a paradigm-shifting influence on bioconjugation chemistry, fundamentally changing the way biomolecular medicines are synthesized in the laboratory and clinic.

项目摘要/摘要 三十年来，含有甲酰亚胺的问题与硫醇之间的连接一直是 位点特异性标记抗体的合成。目的地很受欢迎，这种生物缀合的方法 有严重的局限性。基于马来酰亚胺的偶联物在体内显示有限的稳定性，因为它们的琥珀酰亚胺基 硫醚连接可以发生复古迈克尔的反应，导致货物或其其解离 与循环生物分子交换。在核成像和放射免疫疗法的背景下，这种恢复 迈克尔的反应可能导致放射性有效载荷的释放和体内放射线标记 内差的生物分子，导致对健康组织的辐射剂量较高，减少了成像对比度， 和较低的治疗比率。为了规避这一障碍，我们已经开发了一个模块化，稳定和 易于访问的苯氧甲二二氯甲基磺基磺极试剂 - “豆荚”作为基于硫醇的平台 生物缀合。我们已经证明，POD可用于可重复创建同质，良好 定义，高度免疫反应性和高度稳定的放射免疫共轭物，体内远高 与通过传统的基于马来酰亚胺的方法合成的类似探针相比，性能相比。 该提案的重点是该技术的扩展和优化。我们将寻求建造 在我们的初步工作中，通过优化豆荚 - 硫醇结扎和评估的速度和选择性 基于PODS的放射免疫共轭物的体内性能标记了89ZR，177LU，131i和225AC。 特定的目标1（SA1）将集中于第二库库的设计，合成和表征 一代豆荚试剂的总体目标是确定具有最有利的构造 稳定性，可溶性和反应性的结合。具体目标2（SA2）将以评估为中心 使用POD合成的放射免疫共轭物的体内性能。最有希望的第二 来自SA1的生成POD试剂将用于合成89ZR-和177LU标记的放射免疫共轭物， 它们的体内性能将在癌症的小鼠模型中进行评估，并与 使用基于马来酰亚胺的生物结合产生的放射标记抗体。特定的目标3（SA3）将重点放在 开发基于POD的假体基团，用于用131i和131i和 225AC。使用这些假肢合成的131I-和225AC标记抗体的体内性能 将在癌症的鼠模型中评估，并与使用当前的`gold-合成的类似物进行比较 标准策略。我们认为，该提案可能会对临床护理产生重大的近期影响 开发和验证工具，用于合成具有出色体内的高度稳定的放射免疫共轭物 性能，从而改善成像方案，治疗方案和患者结局。此外， 我们认为，这项工作可能对生物缀合化学产生范式转移影响 改变实验室和诊所中生物分子药物合成的方式。