Mitigating ADA Through Site-specific Conjugation Technology

通过位点特异性缀合技术缓解 ADA

• 批准号: 10750703
• 负责人: Samantha Rene Benjamin
• 金额: $ 0.25万
• 依托单位: STATE UNIVERSITY OF NY,BINGHAMTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-08 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10750703
• 关键词: Achievement; Affinity; Agonist; Allergic Reaction; Antibodies; Antibody Response; Antibody-drug conjugates; Antigen-Presenting Cells; Area; Attenuated; Binding; Biological; Biological Assay; CD80 gene; CD86 gene; Cells; Cysteine; Data; Development; Differentiation Antigens; Dose; Drug Delivery Systems; Engineering; Enzyme-Linked Immunosorbent Assay; Enzymes; Exhibits; FDA approved; Fc domain; Fluorescence; Fluorine; Funding; Goals; Hydrophobicity; IgG1; Immune; Immunologic Adjuvants; Immunologic Stimulation; Immunooncology; Incidence; Location; Macrophage; Masks; Measurable; Mediating; Methods; Modeling; Molecular Conformation; Mus; Oregon; Oregon Green 488 carboxylic acid; Patients; Peptides; Pharmaceutical Preparations; Positioning Attribute; Proteins; Publishing; Reaction; Reporting; Risk; Sampling; Site; TLR7 gene; Technology; Therapeutic; Tumor Tissue; United States National Institutes of Health; Visualization; antibody conjugate; biophysical tools; cancer therapy; clinical application; cytotoxic; design; drug clearance; experimental study; glycosylation; imaging system; immunogenicity; interest; live cell imaging; negative affect; prevent; tool; tumor; uptake

Summary: Antibody drug conjugate (ADC) immunogenicity is a growing concern due to the increased interest in immune- stimulating drug delivery technology. A recent study by Novartis demonstrated that 14 out of 14 patients dosed with an experimental immune-stimulatory ADC developed a measurable anti-drug antibody (ADA) response. ADA negatively affects treatment by neutralizing drugs, increasing drug clearance, and causing severe allergic reactions. There is an urgent need for an effective method of conjugating payload to antibodies while minimizing the development and the effects of ADAs. We have developed an ADC bioconjugation technology that employs the conserved Q295 residue (continued funding provided by NIH 1RO1GM140026-O1A1) that we believe will mitigate many of these ADA concerns. The goal of this proposal is to demonstrate that this technology reduces Fc-gamma uptake in antigen presenting cells (APCs) and masks the drugs from binding to ADAs by hiding them in a sterically occluded hydrophobic pocket on the Fc domain. This goal will be achieved through the two aims. Aim #1 focuses on developing NMR and fluorescence-based tools to demonstrate that ADC payloads at the Q295 site are constrained within the sterically occluded IgG1 hydrophobic pocket. To this end we propose conjugating model payloads to the Q295 site using various linkers (such as short alkyl, peptide, and PEG) and determining their proximity to the hydrophobic pocket using two experimental methods: 1) a proximity induced fluorescent quenching assay using Oregon Green 488, due to its ability to self-quench and 2) F-19 protein NMR using a trifluoromethyl probe, due to its high sensitivity and low background level in biological samples. Aim #2 focuses on demonstrating that payloads attached to the sterically occluded Q295 position have reduced ADA binding and reduced potential for eliciting immunogenicity caused by Fc-gamma mediated uptake of ADCs. Achievement of the aims outlined herein will result in an antibody conjugation technology that exhibits a reduced risk of ADA and may be useful for the design of immune-stimulating antibody conjugates.

概括： 抗体药物结合物（ADC）免疫原性是由于对免疫的兴趣增加而越来越多的关注点 刺激药物输送技术。诺华最近的一项研究表明，14例服用的患者中有14个 使用实验性免疫刺激性ADC产生了可测量的抗药物抗体（ADA）反应。 ADA通过中和药物，增加药物清除率并引起严重过敏，对治疗产生负面影响 反应。迫切需要一种将有效载荷与抗体结合的有效方法 最小化ADA的发展和影响。我们已经开发了ADC生物缀合技术 我们使用保守的Q295残留物（NIH 1RO1GM140026-O1A1提供的持续资金） 相信会减轻许多ADA关注。该提议的目的是证明这一点 技术可减少抗原呈现细胞（APC）中的FC-gamma摄取，并掩盖了药物的结合 ADA将其隐藏在FC域上的空间疏水口袋中。将实现这个目标 通过两个目标。 AIM＃1专注于开发NMR和基于荧光的工具，以证明 Q295站点的ADC有效载荷受到封闭的IgG1疏水口袋的约束。对此 结束时，我们建议使用各种接头（例如短烷基， 肽和钉），并使用两种实验方法来确定其邻近疏水口袋的邻近性： 1）由于其自quench的能力 2）F-19蛋白NMR使用三氟甲基探针，由于其高灵敏度和低背景水平 生物样品。 AIM＃2专注于证明附带的有效载荷附加了c295 位置降低了ADA结合和降低由FC-Gamma引起的免疫原性的潜力 介导的ADC摄取。实现此处概述的目标将导致抗体结合 表现出降低ADA风险并且可能对免疫刺激的设计有用的技术 抗体结合物。