Zwitterionic nanogel encapsulation of uricase to evade immune responses

两性离子纳米凝胶封装尿酸酶以逃避免疫反应

• 批准号: 8951361
• 负责人: SHAOYI JIANG
• 金额: $ 19.31万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-15 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8951361
• 关键词: Address; Adopted; Adverse event; Anaphylaxis; Animal Model; Antibodies; Antibody Response; Attention; Biocompatible Materials; Biodistribution; Blood Circulation; Cells; Characteristics; Chemistry; Clinical Trials; Cross Syndrome; Crosslinker; Dendritic Cells; Development; Digestion; Drug Delivery Systems; Encapsulated; Enzyme-Linked Immunosorbent Assay; Epitopes; Free Radicals; Freeze Drying; Future; Gel; Generations; Gout; Harvest; Immune; Immune response; In Situ; In Vitro; Injection of therapeutic agent; Measures; Membrane Proteins; Modeling; Monitor; NanoGel; Particle Size; Peptide Hydrolases; Pharmacologic Substance; Polyethylene Glycols; Polymers; Process; Property; Proteins; Rattus; Resistance; Serum; Source; Structure; Surface; Techniques; Technology; Temperature; Testing; Therapeutic; Transmission Electron Microscopy; Trypsin; United States Food and Drug Administration; Urate Oxidase; Urea; Work; base; cytokine; immunogenic; immunogenicity; in vivo; intravenous injection; light scattering; macrophage; monomer; novel strategies; particle; patient safety; polymerization; public health relevance; research study; response; success; therapeutic protein; uptake; zeta potential

 DESCRIPTION (provided by applicant): One of the major obstacles that impede the wide application of therapeutic protein products is their potential immunological responses, especially for those obtained from non-human sources. These responses decrease the efficacy of the protein and cause adverse events such as anaphylaxis, cytokine-release syndrome, and cross-reactive neutralization of endogenous proteins, all which may threaten patient safety. Currently, the most successful strategy to mitigate immune response to foreign proteins is the surface conjugation of the amphiphilic polymer polyethylene glycol (PEG) to cover the protein surface epitopes in the "PEGylation" process. This surface coverage 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 have demonstrated the repeated administration of PEGylated therapeutics generating anti-PEG antibodies both in animal models and clinical trials, which directly challenges the future of the PEGylation technology. One typical example is the PEGylated version of the highly immunogenic mammalian uricase for the therapy of gout (approved by FDA in 2010), where the high rate of anti-PEG generation after administration has caused extensive attention. We believe there are two shortcomings for the current PEGylation technology: 1) the PEG polymer is immunogenic and 2) PEG forms sparse brush structures that provides inadequate surface coverage due to its existing and available chemistries. Thus, we propose here a poly(carboxybetaine) (pCB) based nanogel encapsulation technique to overcome these problems simultaneously. In prior studies, we have demonstrated that pCB is a biocompatible material with better non-fouling property and less immunogenicity than PEG. Using pCB as a shielding material, the resulting nanogel will provide 100% protein surface coverage. This new approach adopts the same principle as PEGylation in that coverage of surface epitopes can diminish protein immune responses, but utilizes a non-immunogenic base material and provides a more comprehensive surface coverage. Thus, we hypothesize that pCB-coated uricase will have better circulation profiles and less immunogenicity compared to the current PEGylated version. This hypothesis will be addressed in the experiments of the following Specific Aims: 1) Development of uricase-loaded pCB nanogels with high activity; 2) determination of nanogel stability, stealth characteristics and protease resistance; and 3) determination of circulation profiles, biodistribution, and antibody responses. Successful completion of this proposal will culminate in a new approach for the entire protein pharmaceutical field to generate alternatives to the existing, but insufficient PEGylation technology, producing safer and more effective protein therapeutics.

 描述(申请人提供)：阻碍治疗性蛋白质产品广泛应用的主要障碍之一是它们潜在的免疫反应，特别是那些来自非人类来源的免疫反应。这些反应降低了蛋白质的有效性，并导致过敏反应、细胞因子释放综合征和内源性蛋白质的交叉反应中和等不良事件，所有这些都可能威胁患者的安全。目前，缓解外源蛋白免疫应答最成功的策略是在聚乙二醇化过程中通过两亲性聚合物聚乙二醇单分子的表面偶联来覆盖蛋白的表面表位。这种表面复盖策略已被证明在一定程度上降低了对潜在蛋白质的免疫反应，十多种聚乙二醇化蛋白质产品已被食品和药物管理局(FDA)批准。然而，最近的研究表明，在动物模型和临床试验中，重复使用聚乙二醇化治疗药物会产生抗聚乙二醇化抗体，这直接挑战了聚乙二醇化技术的未来。一个典型的例子是用于治疗痛风的高免疫原性哺乳动物尿酸酶的聚乙二醇化版本(FDA于2010年批准)，其中给药后产生的高抗聚乙二醇率引起了广泛关注。我们认为，目前的聚乙二醇化技术有两个缺点：1)聚乙二醇化聚合物具有免疫原性；2)聚乙二醇化聚合物形成稀疏的刷子结构，由于其现有的和可用的化学成分，表面覆盖率不高。因此，我们在这里提出了一种基于聚羧基甜菜碱(PCB)的纳米凝胶封装技术来同时克服这些问题。在以前的研究中，我们已经证明了PCB是一种生物相容性材料，比聚乙二醇有更好的无污染性能和更低的免疫原性。使用多氯联苯作为屏蔽材料，得到的纳米凝胶将提供100%的蛋白质表面覆盖率。这种新方法采用了与聚乙二醇化相同的原理，即表面表位的覆盖可以减少蛋白质的免疫反应，但使用了非免疫原性的基础材料，提供了更全面的表面覆盖。因此，我们假设，与目前的聚乙二醇化版本相比，多氯联苯包裹的尿酸酶具有更好的循环特征和更低的免疫原性。这一假设将在以下特定目标的实验中得到解决：1)开发高活性的尿酸酶负载的多氯联苯纳米凝胶；2)测定纳米凝胶的稳定性、隐形特性和抗蛋白酶能力；以及3)测定循环系统、生物分布和抗体反应。这项提议的成功完成将为整个蛋白质制药领域带来一种新的方法，以产生现有但不足的聚乙二醇化技术的替代品，生产出更安全、更有效的蛋白质疗法。