Designer Drugs: The Emerging Face of Drug Abuse

我们设计并实施了代谢物图谱研究，将感兴趣的化合物与人肝细胞孵育，并通过高分辨率质谱仪(QTOF 5600)鉴定代谢物。与ABSciex和百时美施贵宝的这一合作项目已经产生了六篇已发表的论文的成果，这些论文描述了不同的新合成大麻素(XLR-11、AKB-48、RCS-4、RCS-8、STS-135、PB-22/5F-PB-22和其他几种)的代谢，并为科学界提供了质谱图，以确定这些药物的用途。目前，新的合成阿片类药物AH-7921和另外三种合成大麻素AB-FUBINACA、AB-PINACA/5F-AB-PINACA正在进行研究。自2014年春季以来，我们还在第二台仪器Thermo QExactive上建立了代谢鉴定研究的分析程序，目前正在处理四种合成卡西酮α-PVT、α-PBP、4-MeO-α-PVP和PV8。作为对人类肝细胞研究的补充，我们设计并实施了人类肝脏微体实验，以评估新设计药物的代谢稳定性(半衰期)，这将提供化合物代谢速度的初步估计。2014年春，我们与分子发现公司建立了合作关系，分子发现公司通过模拟分子在酶催化腔中的3D对接并考虑分子中的活性部位，提供了代谢物的电子预测软件。评估软件的潜力将有助于推进未来的新陈代谢研究。我们还开发了几种分析方法来在人体标本中识别新的设计药物：在LC-MS/MS(QTrap 5500)上利用图书馆搜索的定性分析验证了29种合成大麻素，并使用QTOF5600使用新的SWATH方法验证了更广泛的合成大麻素。使用LC-MS/MS(QTrap 5500)对53种合成大麻素进行了定量分析，并使用Orbitrap技术(QExactive)对28种卡西酮进行了验证。后一种方法最近被扩展到包括33种卡西尼酮、3种苯丙胺和8种哌嗪。建立了MDPV及其两种代谢物的定量分析方法，研究了多种给药途径下MDPV在大鼠体内的药代动力学。我们的目标是持续监测新的精神活性物质在美国市场和全球市场上的表现，并对流行的新化合物进行代谢物鉴定研究。我们的目标是公布我们的发现，并及时向科学界提供MS/MS光谱，以促进分析方法的发展。我们还计划通过使用单独的CYP450酶进行实验来扩大人类肝脏微生物体实验的范围，并阐明途径和预测相互作用。 3.作为与鲍曼博士合作的一部分，我们正在开发一种合成大麻素AM2201及其代谢物的定量分析方法，该方法将用于确定该化合物在大鼠体内的药代动力学。 4.一个长期目标是准备并最终在人体内使用合成大麻素进行受控给药研究。要做到这一点，必须首先进行临床前药理学和毒性测试，以提交研究新药申请，然后才能提交方案。

We have designed and implemented metabolite profiling studies, in which we incubate the compounds of interest with human hepatocytes and identify metabolites via high-resolution mass spectrometry (QTOF 5600). This cooperation project with ABSciex and Bristol Myers Squibb has produced an outcome of six published papers describing the metabolism of different new synthetic cannabinoids (XLR-11, AKB-48, RCS-4, RCS-8, STS-135, PB-22/5F-PB-22, and several others) and providing mass spectra for the scientific community to identify use of these drugs. Currently, studies on AH-7921, a new synthetic opioid, and three more synthetic cannabinoids AB-FUBINACA, AB-PINACA/5F-AB-PINACA are performed. Since spring 2014, we also established the analytical procedures for the metabolism identification studies on a second instrument, the Thermo QExactive, and are currently processing four synthetic cathinones alpha-PVT, alpha-PBP, 4-MeO-alpha-PVP and PV8. Complementing the human hepatocyte studies we designed and implemented human liver microsome experiments to assess the metabolic stability (half-life) of new designer drugs, which will provide a first estimate of how quickly the compound is metabolized. In the spring of 2014, we instituted a cooperation with Molecular Discovery, who provide software for in silico prediction of metabolites by simulating 3D docking of molecules in the enzymes catalytic cavity and consider reactive sites in the molecule. Evaluating the softwares potential will help advancing future metabolism studies. We also developed several analytical assays to identify new designer drugs in human specimens: Qualitative assays with library search were validated for 29 synthetic cannabinoids on LC-MS/MS (QTrap 5500) and for an even wider range of synthetic cannabinoids with the new SWATH approach using the QTOF5600. Quantitative assays were validated for 53 synthetic cannabinoids using LC-MS/MS (QTrap 5500) and for 28 cathinones using Orbitrap technology (QExactive). The latter method was recently extended to include 33 cathinones, 3 amphetamines and 8 piperazines. A quantitative analytical method for MDPV and two metabolites was developed to determine MDPV pharmacokinetics in rats after multiple administration routes. Our goal is to continually monitor the appearance of novel psychoactive substances on the US market and worldwide markets and to perform metabolite identification studies on prevalent new compounds. The goal is to publish our findings and provide MS/MS spectra to the scientific community in a timely manner to facilitate development of analytical methods. We also plan to extend the scope of the human liver microsome experiments by performing experiments with individual CYP450 enzymes and elucidate pathways and predict interactions. 3. As part of the collaboration with Dr. Baumann we are developing a quantitative assay for the synthetic cannabinoid AM2201 and its metabolites, which will be used to determine pharmacokinetics of this compound in rats. 4. A long-term goal is to prepare and eventually perform a controlled administration study with a synthetic cannabinoid in humans. To do so, preclinical pharmacology and toxicity tests have to be run first to file for an Investigational New Drug Application, then a protocol can be submitted.