Abstract：
BACKGROUND: Alcohol use disorder (AUD) is a debilitating physiological and psychiatric disorder which affects individuals globally. The current pharmacological interventions to treat AUD are limited, and hence there is an urgent need for a novel pharmacological therapy which can be effective and safe across the population.
OBJECTIVE: We aimed to investigate a novel neutral cannabinoid receptor-1 (CB1R) antagonist, AM6527, in several preclinical models of ethanol consumption using male and female C57BL6/J mice.
METHODS: Independent groups of male and female mice were subjected to repeated cycles of drinking in the dark (DID), or intermittent access to alcohol (IAA) procedures. Twenty minutes prior to ethanol access in each procedure, animals were treated with intraperitoneal injections of either 1, 3, and 10 mg/kg of AM6527 or its respective vehicle. Acamprosate (100, 200, 300, and 400 mg/kg) or its respective vehicle was used as a positive control. Separate groups of male mice were subjected to a chain schedule of ethanol reinforcement to gain access to ethanol wherein completion of a fixed interval (FI; 5 min) schedule (link 1: \"Seeking\") was reinforced with continuous access to ethanol (fixed ratio; FR1) for up to 1.8 g/kg (link 2: \"consumption\"). All the animals were treated with 1, 3, and 10 mg/kg of AM6527 or its respective vehicle 20 mins prior to the start of the FI chain of the procedure. Separately, AM6527 was also evaluated in male and female mice undergoing acute ethanol withdrawal following 8 weeks of intermittent or continuous access to 20% ethanol drinking.
RESULTS: In both DID and IAA procedures, AM6527 reduced ethanol consumption in a dose-related manner in both male and female mice. AM6527 produced no tolerance in the DID procedure; mice treated with 3 mg/kg of AM6527 for 3 weeks continuously drank significantly smaller amounts of ethanol as compared to vehicle-treated mice over a period of three DID cycles. Moreover, in the IAA procedure, AM6527 caused an increase in water intake over the 24-h period. Acamprosate transiently reduced ethanol intake in male mice in both the DID and the IAA procedures but failed to produce any significant effect in female mice. AM6527 also produced a decrease in the FI responding (\"ethanol seeking\") in animals trained to self-administer ethanol. Lastly, AM6527 mitigated neurological withdrawal signs, i.e., handling induced convulsions (HIC) in mice undergoing acute ethanol withdrawal.
CONCLUSIONS: Current findings support previous studies with CB1R neutral antagonist in reducing voluntary ethanol intake and seeking behavior. Based on results shown in this work, AM6527 can be developed as a first in class CB1R neutral antagonist to treat AUD in both males and females.
OBJECTIVE: We aimed to investigate a novel neutral cannabinoid receptor-1 (CB1R) antagonist, AM6527, in several preclinical models of ethanol consumption using male and female C57BL6/J mice.
METHODS: Independent groups of male and female mice were subjected to repeated cycles of drinking in the dark (DID), or intermittent access to alcohol (IAA) procedures. Twenty minutes prior to ethanol access in each procedure, animals were treated with intraperitoneal injections of either 1, 3, and 10 mg/kg of AM6527 or its respective vehicle. Acamprosate (100, 200, 300, and 400 mg/kg) or its respective vehicle was used as a positive control. Separate groups of male mice were subjected to a chain schedule of ethanol reinforcement to gain access to ethanol wherein completion of a fixed interval (FI; 5 min) schedule (link 1: \"Seeking\") was reinforced with continuous access to ethanol (fixed ratio; FR1) for up to 1.8 g/kg (link 2: \"consumption\"). All the animals were treated with 1, 3, and 10 mg/kg of AM6527 or its respective vehicle 20 mins prior to the start of the FI chain of the procedure. Separately, AM6527 was also evaluated in male and female mice undergoing acute ethanol withdrawal following 8 weeks of intermittent or continuous access to 20% ethanol drinking.
RESULTS: In both DID and IAA procedures, AM6527 reduced ethanol consumption in a dose-related manner in both male and female mice. AM6527 produced no tolerance in the DID procedure; mice treated with 3 mg/kg of AM6527 for 3 weeks continuously drank significantly smaller amounts of ethanol as compared to vehicle-treated mice over a period of three DID cycles. Moreover, in the IAA procedure, AM6527 caused an increase in water intake over the 24-h period. Acamprosate transiently reduced ethanol intake in male mice in both the DID and the IAA procedures but failed to produce any significant effect in female mice. AM6527 also produced a decrease in the FI responding (\"ethanol seeking\") in animals trained to self-administer ethanol. Lastly, AM6527 mitigated neurological withdrawal signs, i.e., handling induced convulsions (HIC) in mice undergoing acute ethanol withdrawal.
CONCLUSIONS: Current findings support previous studies with CB1R neutral antagonist in reducing voluntary ethanol intake and seeking behavior. Based on results shown in this work, AM6527 can be developed as a first in class CB1R neutral antagonist to treat AUD in both males and females.
摘要：
背景：酒精使用障碍（AUD）是一种使人衰弱的生理和精神障碍，影响全球个体。目前治疗AUD的药物干预措施是有限的，因此，迫切需要一种新的药物疗法，它在整个人群中是有效和安全的。
目的：我们旨在研究一种新型的中性大麻素受体-1（CB1R）拮抗剂，AM6527，在使用雄性和雌性C57BL6/J小鼠的几种临床前乙醇消耗模型中。
方法：对雄性和雌性小鼠的独立组进行在黑暗中重复饮酒（DID）的循环，或间歇性获取酒精（IAA）程序。在每个程序中乙醇进入前二十分钟，用腹膜内注射1、3和10mg/kg的AM6527或其各自的载体治疗动物。使用阿坎酸(100、200、300和400mg/kg)或其各自的载体作为阳性对照。对单独的雄性小鼠组进行乙醇强化链计划以获得乙醇，其中完成固定间隔（FI；5分钟）计划（链接1：“寻找”）并连续获得乙醇（固定比例；FR1），最高可达1.8g/kg（链接2：“消耗”）。在程序的FI链开始之前20分钟,用1、3和10mg/kgAM6527或其各自的载体处理所有动物。分别,在经历8周的间歇性或连续获得20%乙醇饮用后的急性乙醇戒断的雄性和雌性小鼠中还评估了AM6527。
结果：在DID和IAA程序中，AM6527以剂量相关的方式减少雄性和雌性小鼠的乙醇消耗。AM6527在DID程序中没有产生耐受性;在三个DID周期内,与媒介物处理的小鼠相比,用3mg/kgAM6527处理3周的小鼠连续饮用显著更少量的乙醇。此外，在IAA程序中，AM6527导致24小时内的水摄入量增加。在DID和IAA程序中,阿坎酸暂时减少雄性小鼠的乙醇摄入,但在雌性小鼠中未能产生任何显著的效果。AM6527还在训练自我施用乙醇的动物中导致FI响应降低（“寻求乙醇”）。最后,AM6527缓解了神经系统戒断症状，即，处理急性乙醇戒断小鼠的诱发惊厥（HIC）。
结论：目前的研究结果支持以前使用CB1R中性拮抗剂在减少自愿乙醇摄入和寻求行为方面的研究。根据这项工作显示的结果，AM6527可以开发为CB1R类中性拮抗剂中的第一个，以治疗男性和女性的AUD。
目的：我们旨在研究一种新型的中性大麻素受体-1（CB1R）拮抗剂，AM6527，在使用雄性和雌性C57BL6/J小鼠的几种临床前乙醇消耗模型中。
方法：对雄性和雌性小鼠的独立组进行在黑暗中重复饮酒（DID）的循环，或间歇性获取酒精（IAA）程序。在每个程序中乙醇进入前二十分钟，用腹膜内注射1、3和10mg/kg的AM6527或其各自的载体治疗动物。使用阿坎酸(100、200、300和400mg/kg)或其各自的载体作为阳性对照。对单独的雄性小鼠组进行乙醇强化链计划以获得乙醇，其中完成固定间隔（FI；5分钟）计划（链接1：“寻找”）并连续获得乙醇（固定比例；FR1），最高可达1.8g/kg（链接2：“消耗”）。在程序的FI链开始之前20分钟,用1、3和10mg/kgAM6527或其各自的载体处理所有动物。分别,在经历8周的间歇性或连续获得20%乙醇饮用后的急性乙醇戒断的雄性和雌性小鼠中还评估了AM6527。
结果：在DID和IAA程序中，AM6527以剂量相关的方式减少雄性和雌性小鼠的乙醇消耗。AM6527在DID程序中没有产生耐受性;在三个DID周期内,与媒介物处理的小鼠相比,用3mg/kgAM6527处理3周的小鼠连续饮用显著更少量的乙醇。此外，在IAA程序中，AM6527导致24小时内的水摄入量增加。在DID和IAA程序中,阿坎酸暂时减少雄性小鼠的乙醇摄入,但在雌性小鼠中未能产生任何显著的效果。AM6527还在训练自我施用乙醇的动物中导致FI响应降低（“寻求乙醇”）。最后,AM6527缓解了神经系统戒断症状，即，处理急性乙醇戒断小鼠的诱发惊厥（HIC）。
结论：目前的研究结果支持以前使用CB1R中性拮抗剂在减少自愿乙醇摄入和寻求行为方面的研究。根据这项工作显示的结果，AM6527可以开发为CB1R类中性拮抗剂中的第一个，以治疗男性和女性的AUD。
