Augmentation of Antioxidant Defenses by Immunotargeting

通过免疫靶向增强抗氧化防御

• 批准号: 7026512
• 负责人: Vladimir R Muzykantov
• 金额: $ 38.69万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7026512
• 关键词: antioxidants; bioengineering /biomedical engineering; catalase; cell adhesion molecules; cytoprotection; disease /disorder model; drug delivery systems; drug design /synthesis /production; drug screening /evaluation; free radical oxygen; gene deletion mutation; hydrogen peroxide; hyperoxia; immunoconjugates; intracellular transport; laboratory mouse; laboratory rat; lung injury; lysosomes; oxidative stress; pharmacokinetics; protein transport; sodium hydrogen exchanger; superoxide dismutase; tissue /cell culture; transfection; vascular endothelium

DESCRIPTION (provided by applicant): Vascular oxidative stress is a key component of many lung diseases, but its treatment is inadequate, in part due to sub-optimal delivery of antioxidants to endothelial cells (EC). Previous studies have shown that: 1) antibodies to Platelet Endothelial Adhesion Molecule (PECAM) permit intracellular delivery of drugs to EC via a novel endocytotic pathway distinct from clathrin- and caveoli-mediated endocytosis; and 2) the antioxidant enzyme (AOE) catalase conjugated with anti-PECAM accumulates in the lungs after IV injection and protects against oxidative lung injury in some animal models. The aim of this translational grant is to enhance the duration and effectiveness of protective effects of this promising strategy to clinically significant levels. We hypothesize that: i) the activity of the conjugates can be prolonged by optimization of their design and by manipulating intracellular trafficking and lysosomal degradation; ii) targeted delivery of additional AOEs to EC will permit enhanced protection from oxidative stress; and iii) pulmonary targeting of optimized AOE conjugates will be protective against hyperoxia. We will test this in the following Specific Aims: 1) define the mechanisms of metabolism of anti-PECAM conjugates in EC. We will test the hypothesis that endocytosis and trafficking involve the cytosolic domain of PECAM, Na+-H+ exchangers (NHE) and rearrangements of cytoskeleton, which can be affected by auxiliary agents to prolong duration of conjugates; 2) design targeting of additional AOE. To enhance protection, tandem SOD/catalase (to detoxify 02- and H202) and 1-CysPrx peroxiredoxin (to detoxify H202 and lipid peroxides) conjugates will be produced. Their composition, activities, EC uptake and protection will be studied in cell culture, while their pharmacokinetics and pulmonary targeting will be determined in naive animals and animals with oxidative lung injury; and 3) evaluate protective effects of the conjugates in animal models. The effectiveness and duration of the effects of new conjugates will be compared and regimens of AOE targeting and mechanisms of protection will be studied in a model of acute EC injury induced by H202 in the pulmonary vasculature in mice. Finally, protection against hyperoxic lung injury by best conjugates will be tested. The overall goal of this proposal is to optimize strategies for vascular AOE targeting, to ultimately initiate translation of this strategy into the clinical domain.

描述（由申请人提供）：血管氧化应激是许多肺部疾病的关键组成部分，但其治疗不足，部分原因是抗氧化剂向内皮细胞（EC）的递送不够理想。先前的研究表明：1)血小板内皮粘附分子（PECAM）抗体允许药物通过一种不同于网格蛋白和小泡蛋白介导的内吞作用的新型内吞途径在细胞内递送到EC；2)抗pecam偶联的抗氧化酶（AOE）过氧化氢酶在静脉注射后在肺内蓄积，对某些动物模型的肺氧化损伤具有保护作用。这项转化资助的目的是将这种有希望的策略的保护作用的持续时间和有效性提高到临床显著水平。我们假设：i)缀合物的活性可以通过优化其设计和操纵细胞内运输和溶酶体降解来延长；ii)有针对性地向EC递送额外的AOEs将增强对氧化应激的保护；iii)优化的AOE偶联物的肺靶向将对高氧具有保护作用。我们将在以下具体目标中对此进行验证：1)确定抗pecam偶联物在EC中的代谢机制。我们将验证内吞作用和运输涉及PECAM的细胞质结构域、Na+-H+交换器（NHE）和细胞骨架重排的假设，这些可能受到辅助剂的影响，以延长偶联物的持续时间；2)额外AOE的设计目标。为了增强保护作用，将产生串联SOD/过氧化氢酶（解毒02-和H202）和1-CysPrx过氧化物还蛋白（解毒H202和脂质过氧化物）偶联物。将在细胞培养中研究它们的组成、活性、EC摄取和保护作用，并在幼稚动物和氧化性肺损伤动物中确定它们的药代动力学和肺靶向性；3)在动物模型上评价结合物的保护作用。在H202致小鼠肺血管急性EC损伤模型中，比较新缀合物的作用效果和持续时间，研究AOE靶向治疗方案和保护机制。最后，将测试最佳结合物对高氧肺损伤的保护作用。本提案的总体目标是优化血管AOE靶向策略，最终将该策略转化为临床领域。