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: 10482416
• 负责人: Chong Cheng
• 金额: $ 19.7万
• 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 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10482416
• 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.

项目摘要 聚合物 - 蛋​​白缀合物引起了极大的兴趣，因为它们在 治疗许多疾病。作为具有高加工性的水溶性聚合物，聚（乙二醇）（PEG）具有 我们被广泛用于蛋白质共轭，并且超过十几种蛋白质已商业化 每年的各种疾病（包括关节炎，癌症，肝炎和痛风等）的临床治疗 出售数十亿美元。但是，PEG可以诱导不利的免疫原性并降低蛋白质生物活性。 Zwitterionic聚合物（ZP）已成为这些方面的钉子的改进，并可能导致 甚至更长的圆时间，但通常线性ZP具有较低的处理性，这会阻碍其应用。 与线性ZPS相比，Zwitterion树状聚合物（ZDS）的可加工性提高，但控制良好的蛋白质 直接由ZDS结合是不可行的。在此R21提案中，我们旨在将ZD与PEG集成 开发ZD修饰的PEG（ZD-PEG）作为蛋白质结合的新型共聚物。中央 假设是ZD-PEG是PEG的改进替代方案，ZD-Pegypated蛋白（ZD-PEG-PTENS） 可以保留蛋白质生物活性，减轻PEG抗原性，并可能具有良好的生物医学对立 特性。根据MPI的完整专业知识，提出了以下两个具体目标：1） 开发ZD-PEG和ZD-PEG-PTENS，以及2）了解ZD-PEG-的生物医学相关特性 ptens。 ZD-PEG每个具有zwitterion（ZW）功能化聚（氨基胺）的ZD单元和PEG 设计具有ω-N-羟基糖酰亚胺（NHS）端子的块。具有变化ZD的ZD-PEG库 一代（G1至G4），ZW类型（羧贝因，硫贝因或磷酸胆碱）和PEG长度（5K和PEG长度 10K）将通过具有炔烃焦点功能的ZD制备来合成10K），然后耦合 ZD具有α-氮化物，ω-NHS钉通过炔烃点击化学。使用猪样尿素作为模型蛋白， 将通过ω-NHS的酰胺化反应制备具有受控卵高密度的ZD-PEG-尿布酶 具有尿酶胺功能的ZD-PEG末端。全面的分析方法将是 用于表征ZD-PEG和ZD-PEG-尿液酶来验证其控制良好的结构。系统 将对ZD-PEG-尿液酶进行财产研究，以深入了解其结构 - 财产关系。 ZD-PEG修饰对溶解性，抗双粘合特性，酶的影响 活动，免疫响应，循环时间，生物分布和尿素酶的毒性将通过 体外和体内研究。具体而言，ZW类型，ZD生成，PEG长度和PEGGYLATION密度如何 将评估影响这些与生物医学相关的特性。拟议的R21研究不仅有望 建立ZD-PEG和ZD-PEG-PTENS的合成方法，但也为其提供了重要的见解 结构依赖性生物医学与相关特性。这些研究将为进一步的基础奠定坚实的基础 开发ZD-PEG-PTENS作为许多临床应用的新疗法。