Development of Esterases for the Treatment of Cocaine Overdose and Abuse

描述（由申请人提供）：古柯酯酶（CocE）是生长在南美洲古柯植物根际的红球菌属细菌的产物。该细菌使用可卡因作为其唯一的碳源和氮源，合成CocE以启动可卡因的代谢。CocE是迄今为止发现的代谢可卡因的最有效的酶（7.2 × 108）。它可以预防和逆转我们的啮齿动物模型中的极端可卡因毒性，并且它有可能被开发成为可卡因毒性作用的第一个有用的拮抗剂。该提案包括继续在可卡因毒性小鼠和大鼠模型中评估这种酶的研究，将其效果与其他基于蛋白质的抗可卡因酶进行比较，并使用各种体外技术，包括聚乙二醇化和插入红细胞，通过降低其抗原性和热敏感性来改善这种酶或其他酶。这些程序的目的是延长酶的作用时间，希望能够获得一些关于如何使这种物质在长期治疗可卡因滥用中有用的见解。

DESCRIPTION (provided by applicant): Cocaine esterase (CocE) is a product of the bacterium Rhodococcus sp. which grows in the rhizosphere of coca plants in South America. The bacterium uses cocaine as its sole source of carbon and nitrogen, synthesizing CocE to initiate metabolism of the cocaine CocE is the most efficient enzyme (7.2 X 108) for metabolizing cocaine yet identified. It can both prevent and reverse extreme cocaine toxicity in our rodent models, and it has the potential to be developed into the first useful antagonist of cocaine's toxic effects. This proposal contains studies to continue to evaluate this enzyme in mouse and rat models of cocaine toxicity, to compare its effects with other protein-based anti-cocaine enzymes, and to use a variety of in vitro techniques, including pegylation and insertion into red blood cells, to improve this or other enzymes by reducing their antigenicity, and heat liability. These procedures are designed to prolong the enzyme's duration of action with the hopeful outcome that some insight will be obtained into how to make this substance useful in the long-term treatment of cocaine abuse.