Immunotherapy to Counteract Lethal Doses of Carfentanil

本提案是对PAR-16-330：“化学威胁对策”的回应 （CounterACT）：治疗性先导化合物的鉴定”，并纳入了 单克隆抗体（mAb）治疗作为急性暴露于强效合成阿片类药物威胁的解决方案， 卡芬太尼从1979年开始，毒品的非法合成被提升到一个非常复杂的阶段， 水平一些吸毒过量的受害者的死亡归因于海洛因，然而，缉获的样本中没有 海洛因，而不是受害者无意中购买“中国白色”（即，α-甲基芬太尼）。芬太尼是一种 一个大的合成镇痛药家族，具有鸦片类药物的所有特性，只是它们的浓度为100- 10，000 比吗啡强一倍芬太尼重新出现的原因与海洛因纯度下降有关， 以提高效力和成本效益的制备。芬太尼可以在一个地方合成 由于它们的效力;一克可以配制成（切割）数千，也许数百万， 剂量要阻止如此少量的纯毒品的传播是极其困难的。因此，虽然 药物管制工作需要继续，治疗合成精神活性药物滥用的工作也需要继续。一 (SPD)类似物卡芬太尼特别令人关注，因为其效力极强（是 吗啡）。它在人类中没有治疗用途，对阿片类药物使用者构成重大死亡风险。另一 问题是卡芬太尼在体内的持久性;药物的半衰期超过纳洛酮，需要 随着时间的推移多次输注纳洛酮并仔细监测，否则会导致再次麻醉。一个 卡芬太尼和瑞芬太尼的雾化混合物在俄罗斯被用作化学战剂， 2002年170名平民死亡一种有希望的减弱SPD作用的治疗策略可以是 在抗体疫苗半抗原缀合物中发现。事实上，这一策略已成功地用于对付可卡因， 尼古丁和冰毒因此，我们计划研究各种抗体流形源于 这些半抗原生物缀合物包括催化抗体;重要的是，每个独特的抗体歧管可以 证明比纳洛酮更有效，因为药物催化剂和可调的半衰期。我们将采取两种方法， 然而，主动和被动疫苗策略，以对抗这种危险的药物，两者都将基于 独特的半抗原设计，在产生具有紧密结合和选择性特异性的抗体中至关重要。 来自后者的抗体将通过在转基因OmniRat中主动接种来获得，并将使用选择 一种方法，其中通过独特的卡芬太尼标记的荧光团与 荧光激活细胞分选和分析。最后，需要一个度量标准来判断这两种类型的价值 治疗卡芬太尼的生理过程的疫苗;我们将研究对卡芬太尼的保护- 在啮齿动物和非人灵长类动物中诱导毒性。针对卡芬太尼的人源化mAb将提供 新的和改进的医疗对策，以打击这种化学威胁的急性毒性。

This proposal is in response to PAR-16-330: “Countermeasures Against Chemical Threats (CounterACT): Identification of Therapeutic Lead Compounds” and incorporates the development of monoclonal antibody (mAb) therapy as a solution for acute exposure to the potent synthetic opioid threat, carfentanil. Beginning in 1979, the illicit synthesis of drugs was elevated to an extraordinarily sophisticated level. A number of overdose victims' deaths were attributed to heroin, however, seized samples contained no heroin, rather the victims were inadvertently buying “China White” (i.e., α-methylfentanyl). The fentanyls are a large family of synthetic analgesics that possess all the properties of the opiates, except they are 100-10,000 times more potent than morphine. Reasons for fentanyl resurfacing are tied to a decline in heroin purity, a need to increase potency and cost-efficient preparation. Designer fentanyl's can be synthesized at a single location and because of their potency; a single gram could be formulated (cut) into many thousand, perhaps millions of doses. Preventing the distribution of such small amounts of a pure drug is exceedingly difficult. Thus, while drug control efforts will need to continue, so will efforts to treat synthetic psychoactive drug (SPD) abuse. One (SPD) analogue, carfentanil, is of particular concern because of its extreme potency (10,000 times that of morphine). It has no therapeutic use in humans, posing a significant mortality risk for opioid users. Another issue arises from the persistence of carfentanil in the body; the half-life of the drug exceeds naloxone, requiring multiple infusions of naloxone over time and careful monitoring or else results in re-narcotization. An aerosolized mixture of carfentanil and remifentanil was used as a chemical warfare agent in Russia, resulting in 170 civilian deaths in 2002. A promising therapeutic strategy for attenuating the effects of SPDs can be found in antibody vaccine hapten conjugate. Indeed this strategy has been successfully used against cocaine, nicotine and methamphetamine. Accordingly, we plan to investigate various antibody manifolds stemming from these hapten bioconjugates including catalytic antibodies; importantly each unique antibody manifold may prove more effective than naloxone due drug catabolism and a tunable half-life. We will take two approaches, active and passive vaccine strategies, to counter this dangerous drug, however, both will be grounded upon unique hapten design that will be critical in producing antibodies with tight binding and selective specificity. Antibodies from the latter will come via active vaccination in transgenic OmniRats, and will use a selection process wherein human antibodies will be secured through unique carfentanil-tagged fluorophore coupled with fluorescent-activated cell sorting and analysis. Finally, a metric will be needed to judge the value of both types of vaccines for treating carfentanil's physiological processes; we will look at protection against carfentanil- induced toxicity in both rodents and nonhuman primates. Humanized mAb's against carfentanil will provide a new and improved medical countermeasure to combat the acute toxicity of this chemical threat.