A FUNCTIONAL CELL BASED ASSAY FOR SYNTHETIC CANNABINOIDS

基于功能细胞的合成大麻素测定

• 批准号: 9979149
• 负责人: Andrew Albert Monte
• 金额: $ 23.11万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2022-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9979149
• 关键词: Accident and Emergency department; Agonist; Antibodies; Arsenicals; Binding; Biological Assay; Biosensor; CNR1 gene; CNR2 gene; Cannabinoids; Categories; Cell Line; Cell membrane; Cells; Cessation of life; Chemicals; Clinical; Colorado; Consumption; Cyclic AMP; Cyclic AMP Receptors; Dangerousness; Designer Drugs; Detection; Disease Outbreaks; Drug Compounding; Drug Screening; Drug usage; Enzymes; Euphoria; Evolution; Exposure to; Fluorescence Resonance Energy Transfer; High Pressure Liquid Chromatography; Human; Immunoassay; Ingestion; Label; Laboratories; Law Enforcement; Licensing; Light; Link; Liquid Chromatography; Mass Spectrum Analysis; Measures; Membrane Potentials; Methods; Molecular; Molecular Conformation; Opioid Receptor; Optics; Output; Parents; Patients; Pharmaceutical Preparations; Pharmacology; Physical Dependence; Potassium Channel; Production; Psychological Dependence; Receptor Activation; Receptor Cell; Reference Standards; Research; Saline; Sampling; Schedule; Signal Transduction; System; Techniques; Testing; Tetrahydrocannabinol; Translating; Universities; Urine; Work; base; bath salts; cannabinoid receptor; clinical effect; cost; drug testing; flexibility; inhibitor/antagonist; innovation; liquid chromatography mass spectrometry; novel therapeutics; receptor; rimonabant; screening; synthetic cannabinoid; synthetic opioid; virtual; voltage sensitive dye

The overall objective of this proposal, A Functional Cell Based Assay for Synthetic Cannabinoids, is to develop a flexible and inexpensive screening assay that can detect all synthetic cannabinoids (SCs). Identification of SC abuse remains difficult for clinicians and law enforcement due to the rapid evolution of SC compounds. There are over 200 SCs known to exist though the most comprehensive clinical assay can only detect 43 of these drugs. In fact, more than 90% of SCs identified in patients seen in the University of Colorado Emergency Department are not detectable through the clinically available assay. High performance liquid chromatography- mass spectrometry methods are far more sensitive, though lack of reference standards, lack of availability, and cost remain a limitation of this drug detection method for the vast majority of clinical and law enforcement applications. New inexpensive drug screens able to detect rapidly evolving SC compounds are necessary. In this proposal, we employ an innovative strategy to detect new synthetic cannabinoids (SCs) through detection of activation of the cannabinoid receptor (CB1). The priorities are to create an assay that (a) will rapidly and cheaply detect binding and/or activation of CB1, (b) will distinguish between THC-level binding/activation and SC-level binding/activation, (c) can be integrated as a pre-screen prior to mass spectrometry confirmation. To achieve these priorities, we will develop and assess three independent detection systems of CB1 agonism via membrane potential change (Aim 1), cAMP production (Aim 2), and receptor conformational change (Aim 3). When completed, this receptor based-strategy can be used for detection of other new psychoactive substances, such as synthetic opioids, and can be optimized for rapid bedside screening of a large number of patients. Since avoidance of detection is a major reason SCs are chosen by users, increased screening will decrease use of these drugs. This technique is a solution for clinical and law enforcement detection of all new psychoactive substances.

本提案的总体目标是开发一种基于功能细胞的合成大麻素测定方法， 这是一种灵活且廉价的筛选方法，可以检测所有合成大麻素（SC）。鉴定 由于SC化合物的快速发展，临床医生和执法人员仍然难以滥用SC。 已知存在超过200种SC，尽管最全面的临床测定仅能检测43种SC。 这些药物。事实上，在科罗拉多大学急诊科发现的患者中， 通过临床可用的检测试剂盒无法检出。高效液相色谱- 质谱分析方法要灵敏得多，尽管缺乏参考标准，缺乏可用性， 对于绝大多数临床和执法部门来说，成本仍然是这种药物检测方法的局限性 应用.新的廉价的药物筛选能够检测快速演变的SC化合物是必要的。 在这项提案中，我们采用了一种创新的策略来检测新的合成大麻素（SC）， 检测大麻素受体（CB 1）的活化。优先事项是创建一种测定法， 快速且廉价地检测CB 1结合和/或活化，（B）将区分THC水平 结合/活化和SC水平结合/活化，（c）可以在质谱分析之前整合为预筛选。 光谱确认为了实现这些优先事项，我们将开发和评估三个独立的检测 通过膜电位变化（Aim 1）、cAMP产生（Aim 2）和受体介导的CB 1激动系统 构象变化（目的3）。 完成后，这种基于受体的策略可用于检测其他新精神活性物质。 药物，如合成阿片类药物，并可以优化用于快速床旁筛选大量的 患者由于避免检测是用户选择SC的主要原因，增加筛查将 减少这些药物的使用。这项技术是一种解决方案，临床和执法检测所有新的 精神活性物质