Formulation and Encapsulation of Enzymic Countermeasures Against Organophosphorus

针对有机磷的酶对策的配制和封装

• 批准号: 8739556
• 负责人: THOMAS J MAGLIERY
• 金额: $ 40.54万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-30 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8739556
• 关键词: Accidents; Acute; Arylesterase; Atropine; Cavia; Chemicals; Chlorpyrifos; Collaborations; Development; Diazepam; Dose; Drug Kinetics; Dursban; Encapsulated; Engineering; Enzyme Stability; Enzymes; FDA approved; Formulation; Freeze Drying; General Practitioners; Glycolates; Goals; Grant; High Density Lipoproteins; In Vitro; Industrial Accidents; Injectable; Institutes; Intramuscular; Intramuscular Injections; Intravenous; Kinetics; Laboratories; Lead; Medical; Methods; Modeling; Ohio; Oils; Organophosphorus Compounds; Oximes; Peptide Hydrolases; Peripheral; Pesticides; Pharmaceutical Preparations; Poison; Poisoning; Polyesters; Polymers; Principal Investigator; Proteins; Reducing Agents; Research Institute; Risk; Sarin; Seizures; Serum; Solubility; Solvents; Standardization; Suspensions; Techniques; Testing; Thermodynamics; Treatment Protocols; Universities; Variant; Work; acute symptom; biomaterial compatibility; bioscavenger; cyclosarin; enzyme activity; evaporation; experimental study; in vitro testing; mass casualty; mutant; nanoemulsion; nanomaterials; nanoparticle; nerve agent; particle; polycaprolactone; programs; protective efficacy; public health relevance

DESCRIPTION (provided by applicant): Organophosphorus (OP) nerve agents (e.g., VX and sarin) and pesticides (e.g., chlorpyrifos/Dursban) are among the most toxic compounds known. Because of the continued use of these pesticides around the world, and because many nations are known to have stockpiles of nerve agents, OP agents represent a significant mass casualty threat from both terrorist attacks and industrial accidents. The treatment regimen for OP poisoning results in incapacitation and does not treat sequelae or effects of lower-dose exposures; it is also difficult in mass casualty scenarios. A solution to this problem is development of an enzymic countermeasure that can rapidly degrade the OP agents before they can exert their full effect. There has been recent progress in the development of catalytic bioscavengers, such as engineered mutants of paraoxoase-1 (PON1), particularly against G-agents. Through a CounterACT U54 grant that is a collaboration among the U.S. Army Medical Institute of Chemical Defense (USAMRICD), The Ohio State University (OSU), and The Weizmann Institute, several highly-active variants of PON1 have been engineered, characterized and tested in guinea pigs for preliminary pharmacokinetics and protective efficacy. Variants with activity against cyclosarin and against a spectrum of G agents have been described, and recent work has identified several variants expected to be effective against several OP pesticides. We propose here preliminary characterization of formulations that would directly lead to injectable enzymes most useful in mass casualty situations. The overall goal of this proposal is to develop a robust, encapsulated PON1 variant formulated to be compatible with intramuscular administration for maximum utility in mass casualty situations. Decreased Cmax, increased t1/2, and enhanced shelf stability are all anticipated to result from nanoparticle formulation of the enzyme. A team of protein biochemists from OSU and encapsulation and nanomaterials experts from Southwest Research Institute (SwRI) have partnered to achieve this goal. This proposal will determine the optimum methods and polymers for encapsulation of several PON1 variants in terms of enzyme activity and stability. The solubility and degradation of the nanoparticles will then be tuned by derivatization with polyethyleneglycols (PEGs) of different sizes. Finally, the best derivatized, encapsulated variants will be formulated for IM administration and tested in vitro. The CounterACT U54 Testing Core at USAMRICD will be used to determine pharmacokinetics, protective efficacy, and post-exposure efficacy for the best IM formulation. These experiments will provide the basis for either a project in the renewal of the U54 Center grant, or an independent U01 directed at final formulation of selected PON1 variants.

描述（由申请人提供）：有机磷（OP）神经毒剂（例如，VX和沙林）和杀虫剂（例如，毒死蜱（chlorpyrifos/Dursban）是已知毒性最大的化合物之一。由于这些杀虫剂在世界各地的持续使用，并且由于许多国家都有神经毒剂的库存，OP制剂代表了恐怖袭击和工业事故的重大伤亡威胁。OP中毒的治疗方案导致失能，并且不治疗低剂量暴露的后遗症或影响;在大规模伤亡情况下也很困难。这个问题的解决方案是开发一种酶对策，该对策可以在OP剂发挥其全部作用之前快速降解OP剂。最近在催化性生物清除剂的开发方面取得了进展，例如对氧化酶-1（PON 1）的工程化突变体，特别是针对G-剂。通过美国陆军防化医学研究所（USAMRICD）、俄亥俄州州立大学（OSU）和魏茨曼研究所（Weizmann Institute）之间的合作，CounterACT U 54基金会已经在豚鼠中设计、表征和测试了PON 1的几种高活性变体，以进行初步的药代动力学和保护功效。已经描述了具有针对环沙林和针对一系列G剂的活性的变体，并且最近的工作已经鉴定了预期对几种OP农药有效的几种变体。我们在这里提出的初步表征配方，将直接导致注射酶最有用的大规模伤亡的情况。该提案的总体目标是开发一种稳健的、封装的PON 1变体，其配制为与肌内给药相容，以在大规模伤亡情况下最大限度地发挥效用。降低的Cmax、增加的t1/2和增强的货架稳定性都预期是由酶的纳米颗粒制剂引起的。来自俄勒冈州立大学的蛋白质生物化学家和西南研究所（SwRI）的封装和纳米材料专家团队合作实现了这一目标。该提案将确定用于封装几种PON 1变体的酶活性和稳定性的最佳方法和聚合物。然后通过用不同尺寸的聚乙二醇（PEG）衍生化来调节纳米颗粒的溶解度和降解。最后，将配制最佳衍生化的包封变体用于IM给药并进行体外测试。USAMRICD的CounterACT U 54测试核心将用于确定最佳IM制剂的药代动力学、保护功效和暴露后功效。这些实验将为更新项目提供基础， U 54中心资助，或独立的U 01，直接用于选定的PON 1变体的最终制剂。