Zwitterionic Dendrimer-modified PEG for Protein Conjugation

用于蛋白质缀合的两性离子树枝状聚合物修饰的 PEG

• 批准号: 10317447
• 负责人: Chong Cheng
• 金额: $ 23.69万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10317447
• 关键词: Affect; Alkynes; Amaze; Amines; Arthritis; Azides; Biocompatible Materials; Biodistribution; Blood Circulation; Chemistry; Clinical Treatment; Coupling; Dendrimers; Development; Disease; Family suidae; Foundations; Future; Generations; Gout; Hepatitis; Immune response; In Vitro; Industry; Lead; Length; Libraries; Malignant Neoplasms; Methodology; Modeling; Modification; PTEN gene; Patients; Phosphorylcholine; Polymers; Preparation; Property; Proteins; Reaction; Reporting; Research; Sales; Solid; Solubility; Structure; Time; Toxic effect; Urate Oxidase; Water; amidation; base; biocompatible polymer; clinical application; copolymer; cytotoxicity; density; design; enzyme activity; ethylene glycol; immunogenicity; improved; in vivo; innovation; insight; interest; novel; novel therapeutics; propargylamine; systemic toxicity; therapeutic protein

PROJECT SUMMARY Polymer-protein conjugates have attracted significant interest because of their critical biomedical applications in treating many diseases. As a water-soluble polymer with high processability, poly(ethylene glycol) (PEG) has been widely used for protein conjugation, and over a dozen PEGylated proteins have been commercialized for the clinical treatment of various diseases (including arthritis, cancer, hepatitis, and gout, etc.), with an annual sale of multi-billion dollars. However, PEG can induce unfavorable immunogenicity and reduce protein bioactivity. Zwitterionic polymers (ZPs) have emerged as improved alternatives for PEG on these aspects and may lead to even longer circulation time, but typically linear ZPs have low processability, which hinders their applications. Zwitterionic dendrimers (ZDs) have improved processability than linear ZPs, but well-controlled protein conjugation directly by ZDs is not feasible. In this R21 proposal, we aim to integrate ZDs with PEGs for the development of ZD-modified PEGs (ZD-PEGs) as novel copolymers for protein conjugation. The central hypothesis is that ZD-PEGs are improved alternatives of PEGs, and ZD-PEGylated proteins (ZD-PEG-PTENs) can retain protein bioactivity, mitigate PEG antigenicity, and possess a range of favorable biomedical-relevant properties. Based on the complementary expertise of MPIs, the following two specific aims are proposed: 1) to develop ZD-PEGs and ZD-PEG-PTENs, and 2) to understand the biomedical-related properties of ZD-PEG- PTENs. ZD-PEGs each having a zwitterion (ZW)-functionalized poly(amidoamine)-based ZD unit and a PEG block with ω-N-hydroxysuccinimide (NHS) terminal are designed. A library of ZD-PEGs with varied ZD generation (G1 to G4), ZW type (carboxybetaine, sulfobetaine, or phosphorylcholine), and PEG length (5K and 10K) will be synthesized by the preparation of ZDs with an alkyne focal functionality, followed by coupling the ZDs with α-azide,ω-NHS PEGs via alkyne-azide click chemistry. Using porcine-like uricase as a model protein, ZD-PEG-uricases with controlled PEGylation density will be prepared through amidation reactions of ω-NHS terminals of ZD-PEGs with amine functionalities of uricase. Comprehensive analytical approaches will be employed to characterize ZD-PEGs and ZD-PEG-uricase to verify their well-controlled structures. Systematic property studies of ZD-PEG-uricases will be performed to achieve an insightful understanding of their structure- property relationships. The effects of ZD-PEG modification on the solubility, anti-biofouling property, enzyme activity, immune response, circulation time, biodistribution, and toxicity of uricase will be investigated through both in vitro and in vivo studies. Specifically, how ZW type, ZD generation, PEG length, and PEGylation density affect these biomedical-related properties will be assessed. The proposed R21 studies promise to not only establish the synthetic methodology for ZD-PEGs and ZD-PEG-PTENs, but also provide critical insights into their structure-dependent biomedical-relevant properties. These studies will lay a solid foundation for the further development of ZD-PEG-PTENs as novel therapeutics for many clinical applications.

项目概要 聚合物-蛋白质缀合物因其在生物医学领域的关键应用而引起了人们的极大兴趣 聚乙二醇（PEG）作为一种具有高加工性能的水溶性聚合物，具有治疗多种疾病的作用。 被广泛用于蛋白质缀合，十多种聚乙二醇化蛋白质已商业化 临床治疗各种疾病（包括关节炎、癌症、肝炎、痛风等），每年 然而，PEG 会引起不利的免疫原性并降低蛋白质的生物活性。 两性离子聚合物 (ZP) 在这些方面已成为 PEG 的改进替代品，并可能导致 甚至更长的循环时间，但通常线性 ZP 的加工性能较低，这阻碍了它们的应用。 两性离子树枝状聚合物 (ZD) 比线性 ZP 具有更好的加工性能，但蛋白质控制良好 直接通过 ZD 缀合是不可行的。在这个 R21 提案中，我们的目标是将 ZD 与 PEG 结合起来。 开发 ZD 修饰的 PEG（ZD-PEG）作为用于蛋白质缀合的新型共聚物。 假设 ZD-PEG 是 PEG 的改进替代品，而 ZD-聚乙二醇化蛋白 (ZD-PEG-PTEN) 可以保留蛋白质生物活性，减轻 PEG 抗原性，并具有一系列有利的生物医学相关特性 基于 MPI 的互补专业知识，提出以下两个具体目标： 1) 开发 ZD-PEG 和 ZD-PEG-PTEN，2) 了解 ZD-PEG- 的生物医学相关特性 PTEN 各自具有两性离子 (ZW) 官能化的基于聚(酰氨基胺) 的 ZD 单元和 PEG。 设计了具有不同 ZD 的 ZD-PEG 库。 世代（G1 至 G4）、ZW 类型（羧基甜菜碱、磺基甜菜碱或磷酸胆碱）和 PEG 长度（5K 和 10K）将通过制备具有炔焦点官能团的 ZD 来合成，然后将 通过炔烃-叠氮化物点击化学与 α-叠氮化物、ω-NHS PEG 形成 ZD，使用猪样尿酸酶作为模型蛋白质， 通过 ω-NHS 的酰胺化反应制备具有受控聚乙二醇化密度的 ZD-PEG-尿酸酶 具有尿酸酶胺功能的 ZD-PEG 末端将成为综合分析方法。 用于表征 ZD-PEG 和 ZD-PEG-uricase 以验证其良好控制的结构。 将进行 ZD-PEG-尿酸酶的性质研究，以深入了解其结构- ZD-PEG修饰对溶解性、抗生物污损性、酶的影响。 将通过以下方式研究尿酸酶的活性、免疫反应、循环时间、生物分布和毒性 具体来说，ZW 类型、ZD 生成、PEG 长度和 PEG 化密度。 拟议的 R21 研究不仅有望对这些生物医学相关特性的影响进行评估。 建立 ZD-PEG 和 ZD-PEG-PTEN 的合成方法，还提供对其的重要见解 这些研究将为进一步的结构依赖的生物医学相关特性奠定坚实的基础。 开发 ZD-PEG-PTEN 作为许多临床应用的新型疗法。