Development and Pharmacology of Novel Lipidic rAHF

新型脂质rAHF的开发及药理学

• 批准号: 7541330
• 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/7541330
• 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; 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; ethylene glycol; immunogenic; immunogenicity; improved; in vivo; inhibitor/antagonist; liver metabolism; molecular assembly/self assembly; mouse model; nano; novel; particle; prevent; programs; protein aggregation; protein complex; 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了蛋白质作为治疗剂的发展。然而，a 常见的并发症是由于抗体反应而降低疗效。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 行政管理。这项提议的总体目标是提高生物制药的治疗效果。 通过开发脂质-蛋白质复合体来降低免疫原性和减少对 蛋白质，从而减少给药频率。(FVIII)提供了一个绝佳的机会 研究这种途径，因为外源性FVIII的使用会导致抑制性疾病的发展 15%-30%的血友病A患者出现抗体反应，并伴有替代治疗。从长远来看 该项目的目标是开发FVIII的脂类复合体，它能积极调节免疫原性和 通行证。在前一个项目期间，合理的方法导致了FVIII的开发- 磷酸丝氨酸(PS)复合体，其对蛋白质的免疫反应在其 在血友病A小鼠体内注射，提高了身体的稳定性。PS与FVIII和FVIII的分子相互作用 利用钙离子开发脂基纳米/微米颗粒，包括脂质体和新型 用于FVIII递送的浓缩和纳米蜗牛结构。在这项建议中，我们建议调查(1) FVIM-PS复合体降低免疫应答的机制(2)低密度脂蛋白的作用 密度受体相关蛋白和von Willebrand因子介导的FVIII和FVIII-PS的清除 (3)抗体反应对FVIII和FVIII-PS复合体清除的影响。这个 旨在了解FVIII-PS复合体药理的研究将在血友病A中进行 小鼠模型和体外抗原提呈细胞，如树突状细胞和T细胞。我们建议 研究免疫系统处理蛋白质抗原的关键步骤，包括 抗原提呈细胞对FVIII的摄取和加工以及随后T细胞的提呈和扩增 细胞。我们将研究PS在调节FVIII免疫原性中的作用(特异性目标1)。这个 由于固有的蛋白质结合，FVIII的药代动力学特性是复杂的，我们将研究 肝脏代谢和免疫系统介导的FVIII和FVIII-PS复合体的处置具体而言 目的研究PS结合和脂质分子组装对药代动力学的影响 参数，如半衰期、曲线下面积和间隙。最后，在具体目标3中，我们将 研究FVIII对抗体应答和处置的影响。从这些研究中获得的结果 将有助于开发最佳剂量以及对疾病和治疗的有效管理。