Towards the Treatment of Cocaine Abuse with a Mutant Bacterial Cocaine Esterase

用突变细菌可卡因酯酶治疗可卡因滥用

• 批准号: 8044725
• 负责人: Remy L. Brim
• 金额: $ 0.18万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-05 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8044725
• 关键词: Advanced Development; American; Animals; Antibodies; Bacteria; Bacterial Proteins; Behavioral; Benzoic Acids; Biological Assay; Blood Pressure; Butyrylcholinesterase; Cardiovascular system; Catalysis; Chronic; Coca; Cocaine; Cocaine Abuse; Cocaine Dependence; Convulsions; Development; Dose; Drug abuse; Enzyme Stability; Enzymes; Goals; Half-Life; Heart Rate; Hour; In Vitro; Injection of therapeutic agent; Location; Mass Spectrum Analysis; Measures; Metabolism; Modeling; Morbidity - disease rate; Mus; Mutate; Mutation; Pharmacological Treatment; Pharmacotherapy; Physiological; Plants; Plasma; Positioning Attribute; Property; Rattus; Reaction; Research; Rewards; Rhodococcus; Schedule; Seizures; Self Administration; Serum; Social Problems; Sodium Channel; Soil; Swiss Mice; Therapeutic; Time; Toxic effect; United States; United States Food and Drug Administration; United States National Institutes of Health; Work; addiction; base; cocaine esterase; cocaine use; ecgonine methyl ester; efficacy testing; enzyme activity; esterase; experience; frontier; immunogenicity; in vivo; monoamine; mutant; protective effect; research study; social; therapy development; thermostability; time use

DESCRIPTION (provided by applicant): The long term goal of this research is to develop a pharmacological treatment for cocaine abuse. Specifically, this proposal will examine the potential of a thermostable mutant of bacterial cocaine esterase to be used as a therapy for cocaine abuse. Cocaine abuse is a widespread problem in the United States. Chronic cocaine use leads to severe physical and social consequences yet currently there is no specific FDA-approved pharmacotherpay for cocaine abuse. This research focuses on the bacterial protein cocaine esterase (CocE), isolated from a Rhodococcus bacterium that lives in the 180C soil surrounding the coca plant. CocE rapidly hydrolyzes cocaine significantly faster than similar enzymes that are in development for cocaine abuse and toxicity. Previous work done by this lab has shown that small doses of cocE are able to effectively block cocaine-induced lethality, convulsions, seizure, as well as increases in blood pressure, heart rate and QT interval. Although the wild type form has an extremely short half-life at 370C in vitro (13 minutes), a new set of mutations, L169K/G173Q, give CocE an in vitro half life of 72 hours. The extended half-life and long duration of action of this mutant makes it a better candidate for an abuse therapeutic than wild-type CocE. Experience with wild-type CocE as well as other cocaine hydrolyzing molecules uniquley positions this group to advance the development of this thermostable mutant of CocE towards a therapy for cocaine abuse. Based on the hypothesis that the long acting mutation of bacterial CocE (L169K/G173Q) may be an effective potential treatment for cocaine addiction, two specific aims are proposed: 1) Determine the efficacy of L169K/G173Q CocE against the lethal and reinforcing effects of cocaine in vivo. The potency and duration of action of L169K/G173Q CocE and wild type cocE against cocaine-induced lethality in NIH Swiss mice will be assessed. The effects of L169K/G173Q CocE on the reinforcing properties of cocaine will be assessed in a rat cocaine self-administration model. 2) Determine the stability and activity of L169K/G173Q CocE in vivo. Enzyme stability will be examined by analyzing the plasma half-life, in vivo rates of catalysis, ex vivo activity and immunogenicity of L169K/G173Q and wild-type CocE. We hope to develop a thermostable mutant bacterial cocaine esterase into a pharmacotherapy for cocaine abuse, a widespread social problem for which there is currently no specific FDA approved treatment. Thermostable cocaine esterase rapidly converts cocaine into inactive products, therefore eliminating or greatly reducing the strong psychotropic and physiological effects of cocaine.

描述（由申请人提供）：本研究的长期目标是开发可卡因滥用的药物治疗方法。具体来说，该提案将研究细菌可卡因酯酶的热稳定突变体用作可卡因滥用疗法的潜力。可卡因滥用是美国的一个普遍问题。长期使用可卡因会导致严重的身体和社会后果，但目前 FDA 还没有批准针对可卡因滥用的具体药物疗法。这项研究的重点是细菌蛋白可卡因酯酶 (CocE)，它是从生活在古柯植物周围 180 摄氏度土壤中的红球菌中分离出来的。 CocE 快速水解可卡因的速度明显快于针对可卡因滥用和毒性而开发的类似酶。该实验室之前所做的工作表明，小剂量的cocE能够有效阻止可卡因引起的致死、抽搐、癫痫发作以及血压、心率和QT间期的增加。尽管野生型形式在 370°C 的体外半衰期极短（13 分钟），但一组新的突变 L169K/G173Q 使 CocE 的体外半衰期为 72 小时。该突变体的半衰期延长且作用持续时间长，使其成为比野生型 CocE 更好的滥用治疗候选者。野生型 CocE 以及其他可卡因水解分子的经验使该小组能够推动 CocE 热稳定突变体的开发，以治疗可卡因滥用。 基于细菌 CocE (L169K/G173Q) 的长效突变可能是可卡因成瘾的有效潜在治疗方法的假设，提出了两个具体目标：1) 确定 L169K/G173Q CocE 对可卡因成瘾的致命和强化作用的功效。体内可卡因。将评估 L169K/G173Q CocE 和野生型 cocE 在 NIH Swiss 小鼠中对抗可卡因诱导致死作用的效力和持续时间。 L169K/G173Q CocE 对可卡因增强特性的影响将在大鼠可卡因自我给药模型中进行评估。 2)测定L169K/G173Q CocE在体内的稳定性和活性。通过分析 L169K/G173Q 和野生型 CocE 的血浆半衰期、体内催化速率、离体活性和免疫原性来检查酶稳定性。 我们希望开发一种耐热突变细菌可卡因酯酶，用于治疗可卡因滥用，这是一个广泛存在的社会问题，目前 FDA 还没有批准的具体治疗方法。耐热可卡因酯酶可迅速将可卡因转化为无活性产物，从而消除或大大减少可卡因的强烈精神和生理作用。