Lateral Flow Assessment and Unanticipated Toxicity of Kratom.
Lateral Flow Assessment and Unanticipated Toxicity of Kratom.
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卡痛的横向流动评估和意外毒性。
DOI：
10.1021/acs.chemrestox.8b00218
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发表时间：
2019
影响因子：
4.1
通讯作者：
Janda,KimD
中科院分区：
文献类型：
--
作者：
Smith,LaurenC;Lin,Lucy;Hwang,CandyS;Zhou,Bin;Kubitz,DianeM;Wang,Huiying;Janda,KimD
的叶的mitragynine speciosiatree（也被称为Kratom）长期以来一直被咀嚼，吸烟，或酿造成茶的人在东南亚国家，如马来西亚和泰国.就在过去的一年，植物Kratom在美国获得普及作为“法律的阿片类药物”和调度它作为滥用药物目前悬而未决.在Kratom中发现的主要生物碱是μ-阿片受体激动剂，mitragynine，其结构包含一个有前途的支架免疫药理学用途.虽然Kratom被认为是一种安全的阿片类药物替代品，但在这里，我们报告了其两种主要精神活性生物碱，mitragynine和7-hydroxymitragynine的LD 50值，与静脉注射时小鼠体内的海洛因相当。鉴于Kratom最近在美国的出现，目前没有可用于执法或卫生专业人员的诊断测试，因此我们试图设计这样的测定。使用Mitragynine作为半抗原设计的起始点，产生具有从生物碱的C9位置延伸的醚接头的半抗原。选择细菌鞭毛蛋白（FliC）作为小鼠主动免疫的载体蛋白，产生32种潜在的单克隆抗体（mAb）用于试验开发。发现了在微摩尔至纳摩尔亲和力范围内的抗米托宁mAb，并检查了它们在产生人类流体样品的方便侧流检测测定中的效用。筛选抗体与mitragynine、7-羟基mitragynine的结合以及在侧流测定中的性能。将两种单克隆抗体亚克隆并进一步纯化，其对mitragynine具有93和362 nM的亲和力。测试条测定优化了缓冲液和尿液中mitragynine的检测截止值为0.5 μg/mL（反映了尿液中药物的预期临床相关水平），随着美国阿片类药物流行病的持续发展，这可能对执法机构和卫生专业人员有益。
The leaves of theMitragynine speciosiatree (also known as Kratom) have long been chewed, smoked, or brewed into a tea by people in Southeastern Asian countries, such as Malaysia and Thailand. Just this past year, the plant Kratom gained popularity in the United States as a “legal opioid” and scheduling it as a drug of abuse is currently pending. The primary alkaloid found in Kratom is a μ-opioid receptor agonist, mitragynine, whose structure contains a promising scaffold for immunopharmacological use. Although Kratom is regarded as a safe opioid alternative, here we report the LD50values determined for its two main psychoactive alkaloids, mitragynine and 7-hydroxymitragynine, as comparable to heroin in mice when administered intravenously. Given Kratom’s recent emergence in the U.S., there is currently no diagnostic test available for law enforcement or health professionals, so we sought to design such an assay. Mitragynine was used as a starting point for hapten design, resulting in a hapten with an ether linker extending from the C9 position of the alkaloid. Bacterial flagellin (FliC) was chosen as a carrier protein for active immunization in mice, yielding 32 potential monoclonal antibodies (mAbs) for assay development. Antimitragynine mAbs in the range of micro- to nanomolar affinities were uncovered and their utility in producing a convenient lateral flow detection assay of human fluid samples was examined. Antibodies were screened for binding to mitragynine, 7-hydroxymitragynine, and performance in lateral flow assays. Two monoclonal antibodies were subcloned and further purified with 93 and 362 nM affinity to mitragynine. Test strip assays were optimized with a detection cut off of 0.5 μg/mL for mitragynine in buffer and urine (reflecting projected clinically relevant levels of drug in urine), which could be beneficial to law enforcement agencies and health professionals as the opioid epidemic in America continues to evolve.