Development and Pharmacology of Novel Lipidic rAHF

新型脂质rAHF的开发及药理学

• 批准号: 6872195
• 负责人: SATHY VENKAT BALU-IYER
• 金额: $ 19.33万
• 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 2007-01-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6872195
• 关键词: biological transport; biotherapeutic agent; blood /lymphatic pharmacology; blood disorder chemotherapy; chemical aggregate; chemical stability; chemical structure function; circular dichroism; coagulation factor VIII; conformation; drug design /synthesis /production; fluorescence spectrometry; hemophilia As; immunologic assay /test; immunoregulation; intermolecular interaction; laboratory rat; lipids; liposomes; macrophage; molecular shape; molecular site; pharmacokinetics; physical property; protein folding; recombinant proteins

DESCRIPTION (provided by applicant): Hemophilia A is an inherited bleeding disorder caused by deficiency of Factor VIII, a blood coagulation protein that functions as a cofactor in the coagulation cascade. It is transmitted as a defect on the X chromosome and affects 1 male in 5000. Recent advances in biotechnology and protein engineering, together with cloning of the gene coding for Factor VIII, have made it feasible to manufacture recombinant human Factor VIII (rAHF). The recombinant preparation promises to be a source of unlimited supply, together with the freedom from the complications of transmission of blood-borne viruses. However, rAHF induces antibody type immune response or inhibitors. It has been shown that immune response for a therapeutic protein is caused by the existence of aggregates, frequent administration and existence of natural antibodies for specific epitope region of the protein. Furthermore, a moderately-short circulating half-time limits the duration of pharmacological effect. The overall goal of this proposal is to develop a lipid-based delivery vehicle for AHF to overcome the immune response and extend circulating halflife (Inhibitor Treatment for Hemophilia using Lipid Protein System, ITHELPS). The rationale to use lipidic protein complexes is three fold: (1) the epitope region of the protein binds to phospholipids, and thus by engineering a lipid-protein complex one could shield the epitope region; (2) the lipidic components can interact with intermediate structures in the unfolding/denaturation pathway, thus stabilizing protein structure during storage and reconstitution, thereby avoiding protein aggregate formation, and (3) the lipid-protein complexes, analogous to liposomal particles, may increase the circulation half-life of the rAHF, thus reducing the frequency of administration. In support of these hypotheses, preliminary studies were carried out to develop rationally a lipidic rAHF. The liposomal vesicles stabilized the protein against aggregation and a prototype formulation with lipidic rAHF was achieved. We propose to characterize the molecular topology, pharmacokinetic behavior and immune response of this novel, rationally developed lipidic rAHF. Further, we propose to investigate liposomal clearance mechanisms, including endocytosis and its role in altering the circulation time and immune response of lipidic rAHF. We also propose to study the epitope specificity of rAHF antibodies and the mechanism of immune response.

描述(申请人提供)：血友病A是一种遗传性出血 因缺乏凝血因子VIII而引起的疾病，凝血蛋白 在凝血级联反应中起辅助因子的作用。它作为一种 X染色体上的缺陷，影响5000名男性中的1名。的最新进展 生物技术和蛋白质工程，以及编码基因的克隆 对于第VIII因子，使制造重组人因子成为可能 Viii(Rahf)。这种重组制剂有望成为无限的来源 供应，以及免于传播的并发症 血液传播的病毒。然而，rahf可诱导抗体型免疫反应或 抑制剂。已有研究表明，一种治疗性蛋白质的免疫反应 由于集合体的存在、频繁的管理和 针对蛋白质特定表位区域的天然抗体。此外，a 中短循环半衰期限制药理作用持续时间 效果。这项提议的总体目标是开发一种基于脂质的递送方式 AHF克服免疫反应延长循环半衰期的载体 (使用脂质蛋白系统抑制血友病的治疗，ITHELPS)。这个 使用脂类蛋白质复合体的理由有三个：(1)表位 蛋白质的区域与磷脂结合，从而通过设计一种 脂-蛋白复合体可以屏蔽表位区域；(2)脂类 组件可以与 去折叠/变性途径，从而稳定蛋白质结构 储存和重组，从而避免蛋白质聚集体的形成，以及 (3)脂-蛋白复合体，类似于脂质体颗粒，可增加 RHF的循环半衰期，从而减少了 行政管理。为了支持这些假设，初步研究 为合理开发脂源性RAHF进行了研究。脂质体囊泡 稳定了蛋白质的抗聚集和原型配方 实现了脂源性RHF。我们建议刻画分子拓扑结构， 合理用药后的药代动力学行为和免疫反应 发达的脂源性RHF。此外，我们建议研究脂质体的清除性。 机制，包括内吞作用及其在改变循环时间中的作用 和脂源性RAHF的免疫应答。我们还建议对表位进行研究 抗rahf抗体的特异性及其免疫应答机制。