Mitigating ADA Through Site-specific Conjugation Technology

通过位点特异性缀合技术缓解 ADA

基本信息

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
项目类别：
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

项目摘要

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：由于人们对免疫原性的兴趣日益浓厚，抗体药物偶联物 (ADC) 的免疫原性日益受到关注。刺激药物输送技术。诺华 (Novartis) 最近的一项研究表明，14 名患者中有 14 人服用了使用实验性免疫刺激 ADC 开发出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来对治疗产生负面影响反应。迫切需要一种有效的方法将有效负载与抗体缀合，同时最大限度地减少 ADA 的发展和影响。我们开发了ADC生物共轭技术使用保守的 Q295 残基（由 NIH 1RO1GM140026-O1A1 继续提供资金），我们相信将减轻许多 ADA 担忧。该提案的目标是证明这一点技术减少了抗原呈递细胞 (APC) 中 Fc-gamma 的摄取，并掩盖了药物与通过将 ADA 隐藏在 Fc 结构域上的空间封闭疏水口袋中。这个目标将会实现通过这两个目标。目标#1 专注于开发基于 NMR 和荧光的工具来证明 Q295 位点的 ADC 有效负载被限制在空间封闭的 IgG1 疏水口袋内。对此最后，我们建议使用各种链接器（例如短烷基、肽和 PEG）并使用两种实验方法确定它们与疏水口袋的接近程度： 1) 使用 Oregon Green 488 进行邻近诱导荧光猝灭测定，因为它具有自猝灭能力 2) 使用三氟甲基探针进行 F-19 蛋白质 NMR，因为其灵敏度高且背景水平低生物样本。目标 #2 重点是证明有效载荷附着在空间遮挡的 Q295 上位置减少了 ADA 结合并降低了由 Fc-gamma 引起的免疫原性的可能性介导的 ADC 摄取。实现本文概述的目标将导致抗体缀合该技术可降低 ADA 风险，并可能有助于免疫刺激药物的设计抗体缀合物。