School of Life Sciences, Arizona State University, Tempe, AZ 85287-4501, USA.
School of Life Sciences, Arizona State University, Tempe, AZ 85287-4501, USA; Center of Immunotherapy, Vaccines, and Virotherapy, Biodesign Institute, Arizona State University, Tempe, AZ 85287-4501, USA.
Prog Neuropsychopharmacol Biol Psychiatry. 2020 Aug 30;102:109961. doi: 10.1016/j.pnpbp.2020.109961. Epub 2020 May 6.
Cocaine use disorders include short-term and acute pathologies (e.g. overdose) and long-term and chronic disorders (e.g. intractable addiction and post-abstinence relapse). There is currently no available treatment that can effectively reduce morbidity and mortality associated with cocaine overdose or that can effectively prevent relapse in recovering addicts. One recently developed approach to treat these problems is the use of enzymes that rapidly break down the active cocaine molecule into inactive metabolites. In particular, rational design and site-directed mutagenesis transformed human serum recombinant butyrylcholinesterase (BChE) into a highly efficient cocaine hydrolase with drastically improved catalytic efficiency toward (-)-cocaine. A current drawback preventing the clinical application of this promising enzyme-based therapy is the lack of a cost-effective production strategy that is also flexible enough to rapidly scale-up in response to continuous improvements in enzyme design. Plant-based expression systems provide a unique solution as this platform is designed for fast scalability, low cost and the advantage of performing eukaryotic protein modifications such as glycosylation. A Plant-derived form of the Cocaine Super Hydrolase (A199S/F227A/S287G/A328W/Y332G) we designate PCocSH protects mice from cocaine overdose, counters the lethal effects of acute cocaine overdose, and prevents reinstatement of extinguished drug-seeking behavior in mice that underwent place conditioning with cocaine. These results demonstrate that the novel PCocSH enzyme may well serve as an effective therapeutic for cocaine use disorders in a clinical setting.
可卡因使用障碍包括短期和急性病理（例如，过量）和长期和慢性障碍（例如，顽固性成瘾和戒断后复发）。目前尚无有效的治疗方法可以有效降低与可卡因过量相关的发病率和死亡率，也无法有效预防恢复期吸毒者的复发。最近开发的一种治疗这些问题的方法是使用能够迅速将活性可卡因分子分解为非活性代谢物的酶。特别是，合理设计和定点突变将人血清重组丁酰胆碱酯酶（BChE）转化为一种高效的可卡因水解酶，对（-）-可卡因的催化效率大大提高。目前，阻碍这种有前途的基于酶的治疗方法临床应用的一个缺点是缺乏一种具有成本效益的生产策略，该策略还足够灵活，可以根据酶设计的不断改进快速扩大规模。植物表达系统提供了一个独特的解决方案，因为这个平台旨在快速扩展，成本低，并具有进行真核蛋白修饰（如糖基化）的优势。我们将可卡因超级水解酶（A199S/F227A/S287G/A328W/Y332G 的植物衍生形式）指定为 PCocSH，可保护小鼠免受可卡因过量的影响，对抗急性可卡因过量的致命影响，并防止接受可卡因位置条件作用的小鼠中已熄灭的觅药行为的重新出现。这些结果表明，新型 PCocSH 酶在临床环境中可能是治疗可卡因使用障碍的有效方法。
