Development and Pharmacology of Novel Lipidic rAHF

蛋白质工程的进展已经导致蛋白质作为治疗剂的发展。但 常见的并发症是由于抗体反应而导致的功效降低。影响抗体的因素 免疫应答包括蛋白质聚集、蛋白质内的免疫原性序列和免疫应答的频率。 局该提案的广泛目标是提高生物药物的治疗效果 通过开发脂质-蛋白质复合物，其将降低免疫原性并减少 蛋白质，从而降低给药频率。因子VIII（FVIII）提供了一个极好的机会， 研究此类方法，因为给予外源性凝血因子VIII会导致抑制性的发展 15-30%的血友病A患者出现抗体应答，使替代治疗复杂化。长期 该项目的目标是开发能积极调节免疫原性的FVIII免疫复合物， 间隙在上一个项目期间，合理的方法导致了FVIII的开发- 磷酸丝氨酸（PS）复合物显示出对蛋白质的免疫应答降低， 注射到血友病A小鼠中，并改善了物理稳定性。PS与FVIII的分子相互作用， 钙离子被用于开发基于脂质的纳米/微米颗粒，包括脂质体和新型纳米/微米颗粒。 用于FVIII递送的浓缩和纳米卷状结构。在本提案中，我们建议调查（1） 降低FVIM-PS复合物介导的免疫应答的机制（2）低浓度的 密度受体相关蛋白和血管性血友病因子介导的FVIII和FVIII-PS清除 抗体应答对FVIII和FVIII-PS复合物清除的影响。的 将在血友病A中开展旨在了解FVIII-PS复合物药理学的研究 小鼠模型和体外用抗原呈递细胞如树突细胞和T细胞。我们建议 研究一般免疫系统处理蛋白质抗原的关键步骤，包括 抗原呈递细胞对FVIII的摄取和加工以及随后的T- 细胞我们将研究PS在调节FVIII（特异性目的1）免疫原性中的作用。的 FVIII的药代动力学性质由于内在蛋白结合而复杂，我们将研究FVIII的药代动力学性质。 肝脏代谢和免疫系统介导的FVIII和FVIII-PS复合物的处置。在特定 目的2：研究PS结合和脂质分子组装对药物动力学的影响 参数如半衰期、曲线下面积和清除率。在具体目标3中，我们将 研究抗体应答和FVIII处置的影响。从这些研究中获得的结果 将有助于开发疾病和治疗的最佳剂量和有效管理。

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.