Worob Adam, Wenthur Cody J
Pharmaceutical Sciences Division, School of Pharmacy, University of Wisconsin-Madison, Madison, WI 53705, USA.
Vaccines (Basel). 2022 Aug 4;10(8):1253. doi: 10.3390/vaccines10081253.
Synthetic cannabinoid receptor agonists (SCRAs) are compounds that mimic the pharmacology of the psychoactive components in cannabis. These compounds are structurally diverse, inexpensive, commercially available, and difficult to identify with modern analytical methods, making them highly accessible for recreational use. Suspected SCRA toxicity, which can present with a breadth of cardiovascular, gastrointestinal, and neurological disturbances, is currently addressed through symptom management followed by a toxicological screening that often occurs long after patient discharge. Here, we report the development of four cross-reactive anti-SCRA bioconjugate vaccines as a platform for developing improved diagnostic and therapeutic interventions against SCRA intoxication, using SCRA-resembling small molecule haptens that combine common subregional motifs occurring within and across different generations of SCRA molecules. Using a combination of multiplexed competitive ELISA screening and chemoinformatic analyses, it was found that the antibodies resulting from vaccination with these bioconjugates demonstrated their ability to detect multiple SCRAs with a Tanimoto minimum common structure score of 0.6 or greater, at concentrations below 8 ng/mL. The scope of SCRAs detectable using these haptens was found to include both bioisosteric and non-bioisosteric variants within the core and tail subregions, as well as SCRAs bearing valine-like head subregions, which are not addressed by commercially available ELISA screening approaches. Vaccination with these bioconjugates was also found to prevent the changes in locomotion and body temperature that were induced by a panel of SCRAs at doses of 1 and 3 mg/kg. Further refinement of this genericized hapten design and cross-reactivity-prioritizing approach may enable the rapid detection of otherwise cryptic SCRAs that arise during overdose outbreaks, and could ultimately lead to identification of monoclonal antibody species applicable for overdose reversal.
合成大麻素受体激动剂(SCRAs)是一类模仿大麻中精神活性成分药理作用的化合物。这些化合物结构多样、价格低廉、可在市场上买到,并且难以用现代分析方法鉴别,这使得它们极易被用于娱乐用途。疑似SCRAs中毒可表现为广泛的心血管、胃肠道和神经功能紊乱,目前通过症状管理以及通常在患者出院很久之后才进行的毒理学筛查来处理。在此,我们报告了四种交叉反应性抗SCRAs生物共轭疫苗的研发情况,以此作为开发针对SCRAs中毒的改进诊断和治疗干预措施的平台,所使用的类似于SCRAs的小分子半抗原结合了不同代SCRAs分子内部和之间出现的常见亚区域基序。通过多重竞争性ELISA筛选和化学信息学分析相结合的方法,发现用这些生物共轭物接种疫苗产生的抗体能够在浓度低于8 ng/mL时,以Tanimoto最小共同结构得分0.6或更高来检测多种SCRAs。发现使用这些半抗原可检测的SCRAs范围包括核心和尾部亚区域内的生物电子等排体和非生物电子等排体变体,以及带有缬氨酸样头部亚区域的SCRAs,而市售ELISA筛选方法无法检测这些变体。还发现用这些生物共轭物接种疫苗可预防一组剂量为1和3 mg/kg的SCRAs所诱导的运动和体温变化。对这种通用化半抗原设计和交叉反应性优先排序方法的进一步优化,可能使我们能够快速检测过量用药暴发期间出现的其他隐匿性SCRAs,并最终可能导致鉴定出适用于逆转过量用药的单克隆抗体种类。