Development of Long-acting Cocaine Hydrolase as a Treatment for Cocaine Abuse

 描述(申请人提供)：可卡因滥用是直接或间接影响大多数社区和家庭的主要公共卫生问题。目前还没有FDA批准的针对可卡因成瘾或过量的药物。可卡因滥用造成的灾难性医疗和社会后果使开发一种反可卡因药物成为高度优先事项。加速可卡因代谢，通过与可卡因代谢的主要途径相似的途径--血浆中人丁酰胆碱酯酶(BChE)催化可卡因的水解--产生生物活性不活跃的代谢物，被认为是治疗可卡因过量和成瘾的最有效的策略。由于野生型BChE对自然产生的(-)-可卡因的催化效率(kcat/Km)很低(kcat=4.1min，Km=4.5µM)，我们最近设计并发现了一组BChE-1突变体，称为可卡因水解酶(CocHs)，与野生型BChE相比，对(-)-可卡因的催化效率至少提高了1000倍。我们发现的第一种获得专利的可卡因的体内和临床数据证明了酶疗法在可卡因滥用方面的前景。除了提高疗效外，我们最近设计、发现和获得专利的新可卡因实体不仅对(-)-可卡因具有显著更高的催化效率，而且具有更长的生物半衰期。建立在我们成功合理设计和发现高效可卡因及其长效形式(LAF或COCH-LAF)的基础上，本次研究将专注于进一步优化、制造工艺开发、配方开发和临床前测试，将最有希望的COCH-LAF作为治疗可卡因成瘾的新候选治疗药物。其具体目标是：(1)优化不仅对(-)-可卡因具有高催化效率而且生物半衰期长的有前景的Coch-LAF；(2)测试有希望的Coch-LAF实体大规模表达的可行性；(3)为目标2中选择的最有希望的Coch-LAF开发大规模生产工艺；(4)利用AIM 3生产的Coch-LAF材料，在动物模型中表征最有希望的Coch-LAF的详细药理和毒理学特征。本研究中开发的最有希望的Coch-LAF实体有望作为一种新的外源性酶高效和安全地适用于治疗人类可卡因成瘾的每月给药计划。这项调查将使最好的COCH-LAF实体为cGMP(当前良好制造实践)蛋白质制造、使能研究新药(IND)和后续临床试验做好准备。因此，这项调查将推动一项有希望的治疗 候选人更接近FDA批准的可卡因成瘾治疗。

 DESCRIPTION (provided by applicant): Cocaine abuse is a major public health problem that directly or indirectly affects most communities and families. There is still no FDA-approved medication specific for cocaine addiction or overdose. Disastrous medical and social consequences of cocaine abuse have made the development of an anti-cocaine medication a high priority. Accelerating cocaine metabolism that produces biologically inactive metabolites via a route similar to the principal cocaine-metabolizing pathway-cocaine hydrolysis catalyzed by human butyrylcholinesterase (BChE) in plasma-is recognized as the most efficient treatment strategy for cocaine overdose and addiction. Since the catalytic efficiency (kcat/KM) of wild-type BChE against the naturally occurring (-)-cocaine is low (kcat = 4.1 min and KM = 4.5 µM), we have recently designed and discovered a set of BChE -1 mutants, known as cocaine hydrolases (CocHs), with at least 1,000-fold improved catalytic efficiency against (- )-cocaine compared to wild-type BChE. In vivo and clinical data for the first one of our discovered and patented CocHs demonstrate the promise of enzyme therapy for cocaine abuse. In addition to improved efficacy, our recently designed, discovered, and patented new CocH entities have not only significantly higher catalytic efficiency against (-)-cocaine, but also possess much longer biological half-lives Built on our success in rational design and discovery of the highly efficient CocHs and their long-acting forms (LAFs or CocH-LAFs), this investigation will focus on further optimization, manufacturing process development, formulation development, and preclinical testing of the most promising CocH-LAF as a novel therapeutic candidate for cocaine addiction treatment. The specific aims are: (1) to optimize a promising CocH-LAF which has not only a high catalytic efficiency against (-)-cocaine, but also a long biological half-life; (2) to test feasibility of lage- scale expression of promising CocH-LAF entities; (3) to develop large-scale manufacturing processes for the most promising CocH-LAF selected in Aim 2; (4) to characterize the detailed pharmacology and toxicology profiles of the most promising CocH-LAF in animal models with the CocH-LAF material produced in Aim 3. The most promising CocH-LAF entity developed in this investigation is expected to be highly effective and safe as a novel exogenous enzyme suitable for a monthly dosing schedule for treatment of cocaine addiction in humans. This investigation will make the best possible CocH-LAF entity ready for the cGMP (current Good Manufacturing Practices) protein manufacturing, Investigational New Drug (IND)-enabling studies, and subsequent clinical trials. Thus, this investigation will move a promising therapeutic candidate closer toward FDA approval for cocaine addiction treatment.