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）：这项研究的长期目标是开发可卡因滥用的药理治疗方法。具体而言，该建议将检查细菌可卡因酯酶的热稳定突变体的潜力，以用作可卡因滥用的治疗。在美国，可卡因滥用是一个普遍的问题。慢性可卡因使用会导致严重的身体和社会后果，但目前尚无针对可卡因滥用的FDA批准的药物疗法。这项研究的重点是从可可菌细菌中分离出的细菌蛋白可卡因酯酶（COCE），该细菌生活在可口可乐植物周围的180c土壤中。 CoCE迅速水解可卡因的速度要比与可卡因滥用和毒性开发的类似酶快得多。该实验室所做的先前工作表明，一小剂量的COCE能够有效阻止可卡因引起的致死性，抽搐，癫痫发作以及血压，心率和QT间隔的增加。尽管野生型形式在370℃的体外（13分钟）时具有极短的半衰期，但新的突变（L169K/G173Q）为Coce提供了72小时的体外半寿命。与野生型Coce相比，这种突变体的延长半衰期和长时间的作用使其成为虐待治疗的候选者。具有野生型COCE以及其他可卡因水解分子Uniquley位置的经验，以推动COCE热稳定突变体的发展，以朝着可卡因滥用的治疗。 基于以下假设：细菌COCE的长作用突变（L169K/G173Q）可能是可卡因成瘾的有效潜在治疗方法，提出了两个具体的目的：1）确定L169K/G173Q COCE对可卡因在Vivo In Vivo中的致死和增强作用的效果。将评估L169K/G173Q COCE和野生型COCE对可卡因诱导的NIH瑞士小鼠致死性的效力和作用持续时间。 L169K/G173Q COCE对可卡因增强特性的影响将在大鼠可卡因自我管理模型中进行评估。 2）确定体内L169K/G173Q COCE的稳定性和活性。通过分析血浆半衰期，在体内催化速率，体内活性和L169K/G173Q和野生型COCE的免疫原性来检查酶稳定性。 我们希望将可卡因可卡因可卡因酯酶开发为可卡因滥用的药物疗法，这是一种广泛的社会问题，目前尚无特定FDA批准的治疗方法。可卡因可卡因酯酶迅速将可卡因转化为非活性产物，因此消除或大大降低了可卡因的强烈精神和生理作用。