Cocaine Degrading Enzymes From Pest Insects Of Coca Plants

古柯植物害虫的可卡因降解酶

基本信息

批准号：
7739603
负责人：
Howard H Gu
金额：
$ 22.5万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-01 至 2011-07-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Cocaine is a powerful psycho stimulant and one of the most addictive drugs of abuse. Despite decades of efforts, no effective pharmacological treatments have been developed for cocaine addiction or acute cocaine toxicity. There are two main approaches in developing the anti-cocaine treatments. 1) pharmacodynamic interventions: searching for compounds that inhibit cocaine binding to its targets, which have not resulted in effective drugs yet; 2) pharmacokinetic interventions: strategies to limit or reduce the amount of cocaine available to act on its target proteins, such as the development of cocaine binding antibodies and improved cocaine degrading enzymes. The antibodies could be overwhelmed by large doses of cocaine while each enzyme protein can degrade many cocaine molecules. Significant progress has been made in improving the relative slow endogenous human enzymes by specific and random mutagenesis and by computation guided mutagenesis. In addition, a very active bacterial esterase has been cloned that hydrolyzes cocaine faster than any other known enzymes. However, this enzyme has no homology to the human enzymes and introducing this bacterial protein itself in the human body may cause severe immune responses. Therefore, we propose a strategy that takes advantage of the evolution power of nature. We propose to clone the cocaine degrading enzymes from Eloria Noyesi, the main insect pest of coca plants from which cocaine is extracted. These insects ingest large quantity of cocaine in its diet. The blood cocaine level in the feeding larvae is quite low and most of the ingested cocaine is degraded, suggesting the presence of super fast cocaine-degrading enzymes that have evolved to deal with the high cocaine diet. The successful cloning and additional studies of the cocaine degrading enzymes from Eloria should reveal the structural insight and catalytic mechanism of super efficient cocaine hydrolysis, which may provide novel ideas on how to modify and improve the human enzymes. Such ideas, combined with studies from other labs, may lead to further progress in engineering an ideal enzyme. Any significant improvement in the efficiency of the cocaine-degrading enzyme will make a big impact on the efficacy of an enzyme-based treatment of cocaine abuse. PUBLIC HEALTH RELEVANCE: We propose to clone the super efficient cocaine-degrading enzymes from Eloria Noyesi, the main insect pest of coca plant E. coca from which cocaine is extracted. The successful cloning and further studies of these enzymes may reveal the mechanistic insight of super efficient cocaine hydrolysis and thus provide novel ideals to further improve the human enzymes, which may eventually lead to effective and specific treatments for cocaine addiction and toxicity.

描述（由申请人提供）：可卡因是一种强大的心理刺激物，也是最令人上瘾的虐待药物之一。尽管做出了数十年的努力，但尚未针对可卡因成瘾或急性可卡因毒性开发有效的药理治疗。开发抗可卡因治疗方法有两种主要方法。 1）药效干预措施：寻找抑制可卡因与其靶标结合的化合物，这些化合物尚未导致有效的药物； 2）药代动力学干预措施：限制或减少可卡因量的策略，可用于作用于其靶蛋白，例如可卡因结合抗体的发展和改善可卡因降解酶。大剂量的可卡因可能使抗体不知所措，而每种酶蛋白会降解许多可卡因分子。通过特定和随机诱变以及计算引导的诱变，在改善相对慢的内源性人酶方面取得了重大进展。此外，克隆了一种非常活跃的细菌酯酶，该酯酶的水解可卡因比任何其他已知酶都快。但是，该酶与人体酶没有同源性，并在人体中引入这种细菌蛋白本身可能会引起严重的免疫反应。因此，我们提出了一种利用自然进化力量的策略。我们建议将可卡因从Eloria Noyesi（可卡因的主要害虫）中克隆起来，这是可卡因的主要害虫。这些昆虫在饮食中摄入了大量可卡因。喂养幼虫中的血液可卡因水平很低，大多数摄入的可卡因都会降解，这表明存在超快速可卡因降解的酶，这些酶已经演变为处理高可卡因饮食。可卡因降解酶的成功克隆和其他研究应揭示超有效可卡因水解的结构洞察力和催化机制，这可能会提供有关如何修饰和改善人类酶的新思想。这些想法与其他实验室的研究相结合，可能会导致工程理想酶的进一步进步。可卡因降解酶效率的任何显着提高都将对基于酶滥用的酶治疗的疗效产生重大影响。公共卫生相关性：我们建议从Eloria Noyesi（可口可乐E.可卡因E.可卡因的主要虫害）中克隆超高效的可卡因降解酶。这些酶的成功克隆和进一步的研究可能揭示了超高效可卡因水解的机械洞察力，因此提供了新的理想，以进一步改善人类酶，这最终可能导致可卡因成瘾和毒性的有效和特定的治疗方法。