Zwitterionic Polypeptide-Protein Conjugation for the Safe and Efficient Delivery of Therapeutic Enzymes

用于安全有效递送治疗酶的两性离子多肽-蛋白质缀合

• 批准号: 10264241
• 负责人: SHAOYI JIANG
• 金额: $ 12.9万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-16 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10264241
• 关键词: Acute; Address; Adverse event; Alanine; Anaphylaxis; Animal Model; Antibodies; Antibody Response; Antigen-Presenting Cells; Binding Proteins; Biological Assay; Biomimetics; Blood Circulation; Cells; Chronic; Circular Dichroism; Clinic; Clinical Trials; Development; Drug Delivery Systems; Drug Kinetics; Enzyme-Linked Immunosorbent Assay; Enzymes; Epitopes; Formulation; Future; Generations; Glutamic Acid; Gout; Half-Life; Haptens; Human; Hydrophobicity; Hyperuricemia; Immune; Immune response; In Vitro; Infusion procedures; Injections; Intravenous; Keyhole Limpet Hemocyanin; Kidney; Liver; Lysine; MALDI-TOF Mass Spectrometry; Measures; Membrane Proteins; Modeling; Modification; Molecular Weight; Monitor; Mus; Peptides; Performance; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Pharmacodynamics; Phenotype; Polyacrylamide Gel Electrophoresis; Polyethylene Glycols; Polymers; Process; Production; Proline; Property; Proteins; Reaction; Refractory; Serine; Serum; Sodium Dodecyl Sulfate; Source; Surface; Technology; Testing; Therapeutic; Time; Toxic effect; United States Food and Drug Administration; Urate; Urate Oxidase; Vacuole; Work; amphiphilicity; base; cross reactivity; cytokine; cytokine release syndrome; density; enzyme structure; hydrophilicity; immunogenic; immunogenicity; improved; in vivo; light scattering; mouse model; novel strategies; patient safety; pharmacodynamic model; polypeptide; success; therapeutic enzyme; therapeutic protein

Zwitterionic Polypeptide-protein Conjugation for the Safe and Efficient Delivery of Therapeutic Enzymes PROJECT SUMMARY One of the major obstacles that impede the wide application of therapeutic protein products is their potential immunological response, especially for those obtained from non-human sources. Currently, the most successful strategy to mitigate immune response induced by foreign proteins is “PEGylation”, i.e., to shield the protein surface epitopes with polyethylene glycol (PEG). This surface conjugation strategy has been shown to decrease to some extent immune responses to the underlying protein and more than ten PEGylated protein products have been approved by the Food and Drug Administration (FDA). However, recent studies both in animal models and clinical trials have demonstrated the presence of induced anti-PEG antibodies after repeated administrations and pre-existing anti-PEG antibodies, which directly challenge the future of this PEGylation technology. We believe that there are two shortcomings for the current PEGylation technology: 1) the haptenic character of amphiphilic PEG leading to anti-PEG antibodies and 2) non- degradable character of amphiphilic PEG leading to vacuolation of kidney and liver. Previously, we developed a zwitterionic poly(carboxybetaine) (PCB) protected uricase, which was shown to be capable of maintaining protein bioactivity, reducing the immunogenicity of encased proteins and extending their circulation time without induced anti-polymer Abs. Extended from the zwitterionic concept, poly(EK) (PEK), a zwitterionic polypeptide comprising of alternative lysine (K) and glutamic acid (E), has all properties of zwitterionic PCB and is also degradable to promote its elimination from the body while PEG or PCB is not. Thus, PEK is a great alternative beyond PEGylation, but has never been studied for its application to drug delivery. The purpose of this R21 proposal is to explore the potential of degradable zwitterionic poly(EK) (PEK)-protein conjugates. As a degradable version of PCB, PEK is anticipated to provide similar benefits as PCB to its modified protein drugs while possessing excellent biodegradability. Our preliminary results showed that PEK did not induce any anti-PEK antibodies even when it was conjugated onto highly-immunogenic keyhole limpet hemocyanin (KLH) commonly used to boost the immunogenicity of haptens. However, significant anti-PEG antibodies were elicited by PEG-conjugated KLH in the parallel study. In this proposal, uricase, a non-human enzyme with strong immunogenicity, will be used as a protein target. We hypothesize and will confirm though this study that PEK-uricase conjugates will provide a safe and efficient strategy for enzyme delivery by greatly reducing immunogenicity, improving pharmacokinetics (PK), and enhancing pharmacodynamics (PD). Since PEGylation has been widely used in protein therapeutics, successful completion of this project will culminate in a new approach as an alternative to or a replacement over the PEGylation technology, producing safer and more effective enzyme therapeutics.

两性离子多肽-蛋白质缀合 安全有效地输送治疗酶 项目概要 阻碍蛋白质治疗产品广泛应用的主要障碍之一是其潜力 免疫反应，特别是目前从非人类来源获得的反应最多。 减轻外源蛋白诱导的免疫反应的成功策略是“聚乙二醇化”，即屏蔽 这种表面缀合策略已被用于蛋白质表面表位与聚乙二醇（PEG）的结合。 显示在一定程度上降低了对基础蛋白质和十多种聚乙二醇化的免疫反应 然而，最近的研究表明蛋白质产品已获得美国食品和药物管理局（FDA）的批准。 动物模型和临床试验均证明存在诱导性抗 PEG 抗体 经过重复给药和预先存在的抗 PEG 抗体，这直接挑战了未来 我们认为目前的PEG化技术存在两个缺点。 技术：1) 两亲性 PEG 的半抗原特征导致抗 PEG 抗体，2) 非 此前，我们开发了两亲性 PEG 的可降解特性，导致肾脏和肝脏形成空泡。 两性离子聚（羧基甜菜碱）（PCB）保护的尿酸酶，被证明能够维持 蛋白质生物活性，降低包裹蛋白质的免疫原性并延长其循环时间 没有诱导抗聚合物抗体从两性离子概念延伸而来，聚（EK）（PEK），一种两性离子 由赖氨酸（K）和谷氨酸（E）组成的多肽，具有两性离子PCB的所有特性 并且可降解以促进其从体内消除，而 PEG 或 PCB 则不能。因此，PEK 是一种。 聚乙二醇化之外的绝佳替代方案，但从未研究过其在药物输送中的应用。 该 R21 提案的目的是探索可降解两性离子聚 (EK) (PEK) 蛋白的潜力 作为 PCB 的可降解版本，PEK 预计将提供与 PCB 类似的优点。 我们的初步结果表明，PEK 可以修饰蛋白质药物，同时具有优异的生物降解性。 即使将其缀合到高度免疫原性的匙孔帽上，也不会诱导任何抗 PEK 抗体 血蓝蛋白 (KLH) 通常用于增强半抗原的免疫原性，但具有显着的抗 PEG 作用。 在这项平行研究中，尿酸酶（一种非人类）通过 PEG 缀合的 KLH 引发了抗体。 具有强免疫原性的酶将被用作我们捕获并确认的蛋白质靶点。 这项研究表明，PEK-尿酸酶缀合物将大大提供安全有效的酶递送策略 降低免疫原性，改善药代动力学（PK），并增强药效学（PD）。 聚乙二醇化已广泛应用于治疗性蛋白，该项目的成功完成将达到高潮 采用新方法作为聚乙二醇化技术的替代品或替代品，生产出更安全的产品 以及更有效的酶疗法。