Development and Pharmacology of Novel Lipidic rAHF

新型脂质rAHF的开发及药理学

• 批准号: 7341719
• 负责人: SATHY VENKAT BALU-IYER
• 金额: $ 27.14万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7341719
• 关键词: A Mouse; Abbreviations; Antibodies; Antibody Formation; Antigen-Presenting Cells; Antigens; Area Under Curve; Binding; Binding Sites; Blood Circulation; C2 Domain; Calcium ion; Cell Culture Techniques; Choline; Circular Dichroism; Complex; Complication; Dendritic Cells; Development; Dimyristoylphosphatidylcholine; Disease Management; Dose; Drug Formulations; Drug Kinetics; Drug or chemical Tissue Distribution; Encapsulated; Epitopes; Ethanolamines; Ethylene Glycols; Exogenous Factors; Factor VIII; Frequencies; Generations; Goals; Half-Life; Hemophilia A; Immune response; Immune system; In Vitro; Inflammatory Response; Injection of therapeutic agent; Knockout Mice; LDL-Receptor Related Protein 1; Lead; Lipid Binding; Lipids; Lipopolysaccharides; Liposomes; Mediating; Micelles; Modification; Molecular; Molecular Sieve Chromatography; Mus; Particle Size; Particulate; Patients; Pharmacologic Substance; Pharmacology; Phosphatidic Acid; Phospholipids; Phosphoserine; Plasma; Play; Preparation; Process; Property; Protein Binding; Protein Engineering; Proteins; Recombinants; Replacement Therapy; Research Personnel; Reticuloendothelial System; Role; Serine; Structure; Surface; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Therapeutic Agents; Time Factors; Treatment Efficacy; base; cytokine; density; design; dioleoyl-N-(monomethoxypolyethylene glycol succinyl)phosphatidylethanolamine; ethanolamine; ethylene glycol; immunogenic; immunogenicity; improved; in vivo; inhibitor/antagonist; liver metabolism; molecular assembly/self assembly; mouse model; nano; novel; particle; prevent; programs; protein aggregation; receptor; receptor density; recombinant antihemophilic factor VIII; research study; therapeutic protein; uptake; von Willebrand Factor

Advances in protein engineering have led to the development of proteins as therapeutic agents. However, a common complication is the reduction of efficacy due to antibody response. Factors that influence antibody response include protein aggregation, immunogenic sequences within the protein and the frequency of administration. The broad objective of this proposal is to improve therapeutic efficacy of biopharmaceuticals by developing lipid-protein complexes that will reduce immunogenicity and decrease the clearance of the protein, thereby reducing frequency of administration. Factor VIII (FVIII) offers an excellent opportunity to investigate such approaches, as the administration of exogenous FVIII leads to development of inhibitory antibody responses in 15-30% of Hemophilia A patients, complicating replacement therapy. The long term goal of the project is to develop lipidic complexes of FVIII that positively modulate immunogenicity and clearance. During the previous project period, rational approaches led to the development of FVIII- Phosphoserine (PS)complexes that showed reduction in immune response against the protein following its injection in Hemophilia A mice and improved physical stability. The molecular interaction of PS with FVIII and Calcium ions was exploited to develop lipid based nano/micro particulates including liposomes and novel condensed and nano-cochleate structures for FVIII delivery. In this proposal, we propose to investigate (1) the mechanism of reduction in immune response mediated by FVIM-PS complexes (2) the effect of low density receptor related protein and von Willebrand factor mediated clearance of FVIII and FVIII-PS complexes and (3) the effect of antibody response on clearance of FVIII and FVIII-PS complexes. The studies aimed at understanding the pharmacology of FVIII-PS complexes will be carried out in Hemophilia A mice model and in vitro with antigen presenting cells such as dendritic cells and T-cells. We propose to investigate key steps in the processing of protein antigen by the immune system in general, which include uptake and processing of FVIII by antigen presenting cells and subsequent presentation and expansion of T- cells. We will investigate the role of PS in modulating the immunogenicity of FVIII (Specific Aim 1). The pharmacokinetic properties of FVIII are complex due to intrinsic protein binding and we will investigate the disposition of FVIII and FVIII-PS complexes mediated by liver metabolism and immune system. In specific aim 2, we will investigate the effect of PS binding and lipid molecular assemblies on pharmacokinetic parameters such as half-life, area under the curve and clearance. Finally, in specific aim 3, we will investigate the effect of antibody response and disposition of FVIII. The results obtained from these studies will be useful to develop optimal dosing and efficient management of the disease and therapy.

蛋白质工程的进步导致蛋白质作为治疗剂的发展。但是， 常见并发症是抗体反应引起的疗效的降低。影响抗体的因素 反应包括蛋白质聚集，蛋白质内的免疫原性序列和频率 行政。该建议的广泛目标是提高生物制药的治疗功效 通过开发脂质 - 蛋白质复合物，以降低免疫原性并降低清除 蛋白质，从而降低给药频率。 Factor VIII（FVIII）提供了一个绝佳的机会 研究这种方法，因为外源FVIII的给药会导致抑制 15-30％的血友病A患者的抗体反应使替代疗法复杂化。长期 该项目的目的是开发FVIII的脂质复合物，以调节免疫原性和 清除。在上一个项目期间，合理的方法导致了FVIII-的发展 磷酸盐（PS）复合物显示出对蛋白质后免疫反应的降低 血友病A小鼠注射并改善了身体稳定性。 PS与FVIII和FVIII的分子相互作用 利用钙离子开发基于脂质的纳米/微颗粒，包括脂质体和新型 用于FVIII递送的凝结和纳米型结构。在此提案中，我们建议调查（1） FVIM-PS复合物介导的免疫反应减少的机制（2）低 密度受体相关的蛋白质和von Willebrand因子介导的FVIII和FVIII-PS的清除率 复合物和（3）抗体反应对FVIII和FVIII-PS复合物清除的影响。这 旨在了解FVIII-PS复合物的药理学的研究将在血友病A中进行 小鼠模型和与抗原呈递细胞的体外模型，例如树突状细胞和T细胞。我们建议 调查免疫系统处理蛋白质抗原的关键步骤，其中包括 抗原呈现细胞对FVIII的摄取和处理，随后表现和扩展T- 细胞。我们将研究PS在调节FVIII的免疫原性中的作用（特定目标1）。这 FVIII的药代动力学特性由于固有的蛋白结合而变得复杂，我们将研究 肝脏代谢和免疫系统介导的FVIII和FVIII-PS复合物的处置。具体 AIM 2，我们将研究PS结合和脂质分子组件对药代动力学的影响 参数，例如半衰期，曲线下的面积和间隙。最后，在特定的目标3中，我们将 研究抗体反应和FVIII的处置的影响。从这些研究中获得的结果 对于开发疾病和治疗的最佳剂量和有效治疗将非常有用。