Trehalose Glycopolymers to Enhance both Pharmacokinetics and Stability of Therapeutic Proteins

海藻糖糖聚合物可增强治疗性蛋白质的药代动力学和稳定性

• 批准号: 9113714
• 负责人: Heather D Maynard
• 金额: $ 33.17万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2020-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9113714
• 关键词: Acute; Aftercare; Agranulocytosis; Antibody Formation; Area; Biochemical; Biodistribution; Biological Assay; Blood; Blood Circulation; Blood Glucose; Chronic; Clinical; Desiccation; Diabetes Mellitus; Dose; Drug Kinetics; Electron Beam; Enzyme-Linked Immunosorbent Assay; Evaluation; Exhibits; Exposure to; Freeze Drying; Freezing; Goals; Granulocyte Colony-Stimulating Factor; Half-Life; Health; Heat Stress Disorders; Heating; High temperature of physical object; Human; Hyaluronidase; Immune response; Injection of therapeutic agent; Insulin; Label; Life; Light; Measures; Medicine; Methodology; Molecular Weight; Monitor; Mus; Neutropenia; Organ; Patients; Permeability; Pharmaceutical Preparations; Plasma; Polymers; Positron-Emission Tomography; Property; Proteins; Quality of life; Refrigeration; Renal function; Research; Safety; Sampling; Scanning; Side; Sterilization; Stress; Techniques; Technology; Temperature; Testing; Therapeutic; Time; Tissues; Toxic effect; Trehalose; United States Food and Drug Administration; chemical addition; chemotherapy; compliance behavior; cost; dosage; ethylene glycol; glycosylation; granulocyte; immunogenicity; improved; in vitro Assay; in vivo; insulin tolerance; liver function; preclinical efficacy; protein degradation; protein profiling; public health relevance; residence; stressor; sugar; therapeutic protein

 DESCRIPTION (provided by applicant): The long-term objective of the proposed research is to establish preclinical efficacy of a platform technology for the stabilization of proteins therapeutics. Inherent instability of protein therapeutics is a large hindrance and in some cases a barrier to their use. We have developed polymers that contain the sugar trehalose in the side chains. The materials stabilize biologics to high temperatures, room temperature, refrigeration, freezing, lyophilization, and even to electron beam sterilization dosages. Furthermore, we have evidence that the polymer confers favorable pharmacokinetic properties to proteins similar to the widely utilized polymer in medicine, poly (ethylene glycol) or PEG. We propose that trehalose glycopolymers will have comparable pharmacokinetic properties to PEG and yet be significantly better than PEG with regard to stabilization ability. This is important for patient health, quality of life, and to reduce costs. We plan to demonstrate this with three important therapeutics: insulin and granulocyte colony-stimulating factor (G-CSF) and hyaluronidase (HAse). These representative proteins have different sizes and degrees of glycosylation and are important to stabilize. Yet, if this research is successful, the polymers may be broadly applicable to many other therapeutics. To reach our major objectives, three specific aims are proposed. First, we will prepare polymer-insulin conjugates and determine bioactivity before and after subjection to heat stress. It is hypothesized that trehalose glycopolymer-protein will be comparable to PEG-protein with regard bioactivity, yet exhibit superior stability to heat compared to the PEG conjugate and unmodified protein. To test this, conjugates will be synthesized and subjected to increases in temperature. The protein integrity will be studied biochemically to evaluate size, aggregation state and folding. The bioactivities will then be determined with standard in vitro assays and in vivo assays in mice for insulin, G-CSF and HAse. Second, we will study pharmacokinetic properties and immune response of polymer conjugates. The hypothesis is that conjugated trehalose glycopolymers will prolong the blood half-life of proteins similar to PEG and will be non antigenic. To determine this, standard pharmacokinetic and immunogenicity studies will be undertaken and compared to the analogous PEGylated versions. Third, we will investigate biodistribution and toxicity of conjugates. It is hypothesized that conjugated trehalose glycopolymers will be nontoxic in vivo and will be distributed and eliminated from mice similarly to PEG. This will be ascertained by biodistribution studies utilizing positron emission tomography and gamma counting. The acute and chronic toxicity will be studied in mice by hematological analysis, biochemical parameters, and histological evaluation. These studies are critical to evaluate if trehalose glycopolymers are safe and can be explored further to enhance the life-times of proteins both inside and outside of the body. Studies to identify polymers that can replace PEG in conjugates are important and are part of our long-term goal to replace PEG in protein conjugates.

 描述（应用程序提供）：拟议研究的长期目标是建立平台技术以稳定蛋白质治疗的临床前效率。蛋白质治疗的固有不稳定是一个很大的障碍，在某些情况下是其使用的障碍。我们开发了聚合物，这些聚合物在侧链中含有糖海藻糖。材料将生物制剂稳定在高温下，室温，冷藏，冷冻，冻干，甚至是电子束灭菌剂量。此外，我们有证据表明，聚合物对类似于医学中广泛使用的聚合物，聚（乙二醇）或PEG的蛋白质贡献了有利的药代动力学特性。我们建议海藻糖糖聚合物具有与PEG相当的药代动力学特性，但就稳定能力而言，其比PEG明显好。这对于患者健康，生活质量和降低成本很重要。我们计划使用三种重要的治疗剂来证明这一点：胰岛素和粒细胞群刺激因子（G-CSF）和透明质酸酶（HASE）。这些代表性蛋白具有不同的大小和程度的糖基化，对于稳定很重要。但是，如果这项研究成功，聚合物可能广泛适用 为了达到我们的主要目标，提出了三个具体目标。首先，我们将准备聚合物 - 胰岛素结合物，并确定对热应激的征服之前和之后的生物活性。假设海藻糖糖聚合物 - 蛋​​白质与PEG蛋白在生物活性方面相当，但与PEG共轭和未修饰的蛋白相比，对热的稳定性却具有出色的稳定性。为了测试这一点，结合物将合成并约束温度升高。将对蛋白质完整性进行生化研究，以评估大小，聚集状态和折叠。然后，将用标准的体外测定法和在胰岛素，G-CSF和HASE的小鼠中确定生物活性。其次，我们将研究聚合物偶联物的药代动力学特性和免疫学反应。假设是共轭的海藻糖糖聚合物将延长类似于PEG的蛋白质的血液半衰期，并且将是非抗原的。为了确定这一点，将进行标准的药代动力学和免疫原性研究，并将其与类似的pegyped版本进行比较。第三，我们将研究结合物的生物分布和毒性。假设共轭的海藻糖糖聚合物在体内将是无毒的，并且将分布并从小鼠中与PEG相似地消除。使用正电子发射断层扫描和伽马计数的生物分布研究将确​​定这一点。通过血液学分析，生化参数和组织学评估，将在小鼠中研究急性和慢性毒性。这些研究对于评估海藻糖糖聚合物是否安全至关重要，可以进一步探索以增强体内和外部蛋白质的生命时光。鉴定可以替代偶联物中PEG的聚合物的研究很重要，并且是我们替代蛋白质结合物中PEG的长期目标的一部分。