Mechanisms of alcohol and menthol dependent pharmaco-neurological interactions with arecoline, an addictive and toxic areca (betel) nut xenobiotic

项目总结/摘要 精神活性物质提供暂时的欣快感，但也会对人造成多重伤害 身体槟榔（槟榔）是世界上第四大最常见的滥用物质，仅次于酒精，咖啡因， 和尼古丁。虽然安咀嚼是传统上公认的国外，新出现的证据表明，使用 在美国的移民和难民社区。尽管全球流行，但缺乏知识 关于坚果中主要的精神活性和有毒物质槟榔碱是如何代谢的， 如果它的新陈代谢受到其他物质的干扰，就会产生生物学后果。乙醇通常被共同滥用 与AN，并显着他们的联合摄入与更大的患者伤害的原因尚未 测定我们的支持数据表明，乙醇抑制槟榔碱代谢，表明槟榔碱 在血液和脑组织中会发生蓄积-一种非常显著的毒代动力学相互作用。我们有 还发现L-薄荷醇存在于在美国销售的商业化（品牌）AN产品中，我们的 初步研究结果显示薄荷醇阻止槟榔碱水解。总的来说，这一长期目标 该项目旨在改善AN物质滥用者的健康状况。本申请的总体目标是（i） 确认催化槟榔碱水解的人水解酶和组织位点;（ii）表征新发现的 乙醇药物与槟榔碱的相互作用;和（iii）区分槟榔碱的药代动力学特征的变化， 与L-薄荷醇共同摄入后的神经生物学活性，L-薄荷醇是一种在品牌AN中引起关注的物质 产品.中心假设是槟榔碱与乙醇结合时会受到代谢抑制 或L-薄荷醇，导致毒性增加和更大的CNS作用。中心假设将由3个 具体目标：1）确定水解的人酯酶和组织位点的主要贡献 槟榔碱; 2）剖析乙醇对槟榔碱处置的影响; 3）阐明槟榔碱的药理作用。 薄荷醇在商业化AN产品中的神经学意义。这些目标将由一个 在痕量药物分析、羧酸酯酶药物代谢 仿生3D类器官系统、特殊基因敲除小鼠模型和药物成瘾。雄性和雌性小鼠 将在整个目标中使用，以评估性别作为一个生物变量。5年后，我们希望 首次证明槟榔碱主要在肝脏中通过羧酸酯酶-1（CES 1）代谢， 我们预期的一种酶在乙醇存在下催化槟榔碱的酯交换，形成独特的 具有毒理学特性的代谢物。此外，我们预计L-薄荷醇与槟榔碱的组合将引起 槟榔碱在脑组织中的积聚，并改变小鼠的神经生物学特征和精神活动，这是一种全新的 药物-神经相互作用集团的发现将推进对AN成瘾的了解，并产生 为实施减轻这种精神活性物质毒性负担的战略奠定了科学基础。

PROJECT SUMMARY/ABSTRACT Psychoactive substances provide temporary feelings of euphoria, but also elicit multifold harm to the human body. Areca (betel) nut is the 4th most common abused substance worldwide, only behind alcohol, caffeine, and nicotine. Though AN chewing is traditionally recognized abroad, emerging evidence demonstrates use within migrant and refugee communities in the US. Despite global popularity, there is a dearth of knowledge about how the predominant psychoactive and toxic substance in the nut, arecoline, is metabolized and the biological consequences if its metabolism is perturbed by other substances. Ethanol is commonly co-abused with AN, and significantly their combined ingestion is associated with greater patient harm for reasons yet to be determined. Our supporting data demonstrate that ethanol inhibits arecoline metabolism, suggesting arecoline accumulation will occur in the blood and brain tissue – a highly significant toxicokinetic interaction. We have also identified that L-menthol is present in commercialized (branded) AN products sold in the US, and our preliminary findings show menthol deters arecoline hydrolysis. Taken together, the long-term goal of this project is to improve the health of AN substance abusers. The overall objectives in this application are to (i) confirm human hydrolase(s) and tissue site(s) catalyzing hydrolysis of arecoline; (ii) characterize a newfound ethanol drug interaction with arecoline; and (iii) distinguish changes in arecoline's pharmacokinetic profile and neurobiological activity following co-ingestion with L-menthol, a substance of emerging concern in branded AN products. The central hypothesis is that arecoline is subject to metabolic inhibition when combined with ethanol or L-menthol leading to increased toxicity and greater CNS effects. The central hypothesis will be tested by 3 specific aims: 1) Establish the predominate contribution of human esterases and tissue sites that hydrolyze arecoline; 2) Dissect the influence of ethanol on arecoline disposition; and 3) Elucidate the pharmaco- neurological significance of menthol in commercialized AN products. The aims will be accomplished by an interdisciplinary team with varied expertise in trace drug analysis, carboxylesterase drug metabolism, biomimetic 3D organoid systems, special knock-out mouse models, and drug addiction. Male and female mice will be used throughout the aims to assess sex as a biological variable. Ultimately after 5-years, we expect to be the first to demonstrate arecoline is predominantly metabolized in the liver by carboxylesterase-1 (CES1), an enzyme we anticipate catalyzes transesterification of arecoline in the presence of ethanol, forming a unique metabolite with toxicological properties. Further, we anticipate L-menthol combined with arecoline will provoke arecoline buildup in brain tissue and modify the neurobiological profile and psychoactivity in mice, a brand new pharmaco-neurological interaction. Findings in conglomerate will advance insight about AN addiction, and yield a scientific foundation for implementing strategies to lessen this psychoactive substance's toxic burden.