Project 1 - Chemistry & In Vitro Studies of Chinese Herbal Remedies

酒精和药物滥用在美国和世界各地造成严重的医疗，社会和经济问题。大量的努力已被导向开发有效的治疗方法。不幸的是，在过去的40年里，只有三种疗效有限的药物被美国食品和药物管理局批准。可卡因依赖的复杂性和缺乏有效的补救措施，特别是对复发，这往往是 即使在长时间禁欲之后，由于戒断和/或强烈的渴望而引发的，对治疗提出了严重的挑战。一种可能的作用机制是通过kappa阿片受体来改变可卡因的行为效应。由于18世纪和19世纪在中国最初开发的许多草药都是针对鸦片成瘾的，因此项目1将筛选的化学成分 对于κ受体活性，可以发现具有κ激动剂或拮抗剂作用。已经确定的是，可卡因的一些滥用相关作用可以通过κ阿片样物质受体来改变，并且这些作用与κ阿片样物质对丘脑核中多巴胺水平的调节有关。例如，在大鼠中，κ阿片样物质激动剂已显示出减弱可卡因刺激的运动活动、可卡因辨别和可卡因自我给药。此外，κ阿片样物质激动剂也已显示减弱可卡因诱导的中脑核中多巴胺细胞外水平的增加。此外，最近的研究还表明，kappa拮抗剂可能有助于预防阿片类药物和可卡因使用的复发。因此，集中于替代药物疗法的联合努力将具有重要的临床意义，特别是在预防复发方面。 我们选择了两种中草药，YGT（NPI-025）和XJL（NPI-028），因为它们在中国治疗药物滥用方面的有效性已得到证实，并且近年来在我们的实验室中获得了令人鼓舞的科学数据。本研究项目1的具体目的是（1）通过HPLC指纹图谱获得并标准化这两种草药;（2）通过分馏提供纯化组分作为内标物和分子标准物。 （3）将这些标准化的草药和组分分发给研究项目1、2和4，分别进行体外、体内和临床评价;和（4）进行中草药及其组分对μ、δ和κ阿片受体的亲和力的体外筛选，伤害感受素/孤啡肽FQ（N/OFQ）受体和D1和D2多巴胺受体以及它们通过[35 S] GTP γ S结合和腺苷酸环化酶活性的受体功能。在项目2中，将使用各种体内模型对在体外显示阳性结果的草药或组分进行检查。只有体内活性最好的药物才能在项目4中进行临床评价。

Alcohol and drug abuse pose serious medical, social, and economic problems in the United States and around the world. A great deal of effort has been directed toward developing effective therapies. Unfortunately, in the last 40 years, only three medications with limited efficacy have been approved by the U.S. Food and Drug Administration. The complexity of cocaine dependence and the lack of effective remediation, especially for relapse, which is often precipitated by withdrawal and/or intense craving even after prolonged abstinence, poses a serious therapeutic challenge. One possible mechanism of action underlying the modification of cocaine's behavioral effects by herbal remedies is through kappa opioid receptors. Because many herbal remedies that were originally developed during the 18th and 19th centuries in China were targeted at opium addiction, chemical fractions that will be screened in Project 1 for kappa receptor activity may be found to have kappa agonist or antagonist actions. It is well established that some abuse related effects of cocaine can be modified through kappa opioid receptors, and these effects have been related to kappa opioid modulation of dopamine levels in the nucleus accumbens. For example, in rats, kappa opioid agonists have been shown to attenuate cocaine-stimulated locomotor activity, cocaine discrimination and cocaine self-administration. Moreover, kappa opioid agonists have also been shown to attenuate cocaine-induced increases in extracellular levels of dopamine in the nucleus accumbens. In addition, recent studies also indicate that kappa antagonists may be useful in preventing relapse to opioid and cocaine use. Therefore, a consortium effort focusing on alternative pharmacotherapies would have important clinical significance, especially in preventing relapse. We selected two Chinese herbal medicines, YGT (NPI-025) and XJL (NPI-028), for this project because of their proven efficacy in the treatment of substance abuse in China and encouraging scientific data obtained in recent years in our laboratories. The specific aims of this Research Project 1 are (1) to procure and standardize these two herbal medicines by HPLC fingerprinting; (2) to provide purified components by fractionation as internal standards and as molecular tools to elucidate the mechanism of action; (3) to distribute these standardized herbal medicines and fractions to Research Projects 1, 2, and 4 for in vitro, in vivo, and clinical evaluations respectively; and (4) to conduct in vitro screening of Chinese herbal remedies and fractions for their affinity at the mu, delta and kappa opioid receptors, the nociceptin / orphanin FQ (N/OFQ) receptor and D1 and D2 dopamine receptors and for their receptor functions by [35S]GTPgammaS binding and adenylyl cyclase activity. The herbal medicines or fractions that show positive results in vitro will be examined in Project 2 using various in vivo models. Only those with the best in vivo activity will be subiect to clinical evaluation in Project 4.