Development of safe and effective immunotherapeutics for treatment of opioid abuse and overdose

项目摘要 滥用阿片类药物，包括海洛因和合成阿片类药物(如芬太尼)的情况日益严重。 美国的一个问题，它对最近宣布的国家紧急状态负有部分责任 阿片类药物危机。2006至2015年间，与海洛因有关的死亡人数增加了600%以上，其中 海洛因掺杂的合成阿片类药物(例如，“杀手类阿片类药物”)是滥用#造成的大部分死亡原因。 阿片类药物。尽管FDA批准的治疗阿片成瘾的药物已经上市，但阿片成瘾的使用率 这些药物受到副作用、严格的处方指南和可获得性限制的限制。这个 阿片类药物滥用的持续增加创造了对提供持久、安全和成本低的新疗法的迫切需求。 对过量和复发进行有效的干预，但避免与当前成瘾相关的副作用 药物。使用药物特异性抗体(即免疫拮抗剂)进行的免疫药物治疗 阿片类药物活性和防止靶向阿片类药物进入大脑是很有前途的治疗方法 这些药物的副作用比传统的阿片类药物替代治疗要少。在这个SBIR项目中， 我们建议开发一种新的免疫疗法--纳米抗体(例如，单一结构域 对海洛因、芬太尼具有高度特异性和亲和力的抗体)治疗 阿片类药物“过量和滥用。我们预计阿片类药物特定的纳米体会更好 免疫疗法比传统的免疫球蛋白G为基础的药物抗体，因为纳米体 非常健壮，高度抵抗变性，具有超强的组织渗透性，并具有杂交能力 血脑屏障。我们的目标是获得针对阿片类药物滥用的持久和高滴度抗体。 第一阶段的项目将集中在体外方法，以产生高密度的特定纳米体。 与噬菌体展示纳米体库中的“类阿片杀手”的亲和力。在一期工程中，我们将使 阿片类药物的衍生物，使它们可以在高密度的磁性上化学偶联 并分离针对海洛因和芬太尼的高亲和力纳米体。此外，我们还将设计 表达单个单域抗体，制备二价(多价)纳米体或多价 纳米体偶联纳米粒作为靶向两个或两个以上阿片类药物的新疗法 同时。这些纳米抗体将通过以下方式改进目前的药物特异性抗体范例 增加多价性并允许药代动力学定制，同时避免与 内源性抗体受体途径。成功完成一期工程将产生新的 并为开发针对阿片类药物的免疫疗法提供了一个框架 其他滥用药物。在未来的第二阶段，我们将进行临床前的体外和体内啮齿动物 单价和多价纳米体中和治疗作用的药代动力学研究 致命的阿片类药物。

Project Summary The abuse of opioids, which include heroin and synthetic opioids (e.g., fentanyl), is a growing problem in the United States that is partly responsible for the recently declared National Emergency for Opioid crisis. Between 2006 and 2015, the number of heroin-related deaths increased over 600% where heroin laced synthetic opioids (e.g., the “killer opioids”) is responsible most of the deaths from abuse of opioids. Although FDA-approved medications to treat opioid addiction are available, the utilization rate for these medications is limited from side effects, tight prescription guidelines, and restricted availability. The ongoing rise in opioid abuse creates a dire need for new therapy that offers long-lasting, safe, and cost- effective interventions for overdoses and relapses but avoid side effects associated with current addiction medications. Immunopharmacotherapy using drug-specific antibodies (i.e., immunoantagonists) to block opioid drug activity and prevent the target opioids entering the brain are promising treatment approach that have less side-effect than the traditional drug-based opioid replacement treatment. In this SBIR project, we propose to develop a new class of immunotherapeutics-the nanobodies (e.g. the single domain antibodies, sdAbs) that have high specificities and affinities to heroin, fentanyl for the treatment of “killer opioids” overdose and abuse. We expect the opioid-specific nanobodies will be better immunotherapeutics than traditional immunoglobulin G-based drug antibodies because nanobodies are extremely robust, highly resistant to denaturation, have superior tissue penetration, and have ability to cross blood brain barrier. Our goal is to achieve long-lasting and high titer antibodies to opioid drugs of abuse. The phase I project will focus on in vitro methodologies for the generation of specific nanobodies with high affinities to “killer opioids” from phage-display nanobody library. In the Phase I project, we will make derivatives of opioid drugs such that they can be chemically conjugated at high density on magnetic beads, and isolate high affinity nanobodies specific to heroin and fentanyl. Furthermore, we will engineer and express individual single domain antibody to produce bivalent (multivalent) nanobody or multi- nanobody-conjugated nanoparticles as new therapeutic modalities targeting two or more opioids simultaneously. These nanobodies will improve upon current drug-specific antibody paradigms by increasing multivalency and allowing pharmacokinetic customization, while avoiding interactions with endogenous antibody receptor pathways. Successful completion of Phase I project will generate novel opioid-specific nanobodies and provide a framework for the development of immunotherapeutics against other drugs of abuse. In future Phase II, we will perform the pre-clinical in vitro and in vivo rodent pharmacokinetic study of individual nanobodies and multivalent nanobody for therapeutic neutralizations of the killer opioids.