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（例如，芬太尼），是一个不断增长的 美国最近宣布的国家紧急状态的部分原因是， 鸦片危机2006年至2015年间，与海洛因有关的死亡人数增加了600%以上， 掺有海洛因的合成阿片样物质（例如，“致命的阿片类药物”）是滥用阿片类药物造成的大多数死亡的原因。 阿片类药物虽然FDA批准的治疗阿片类药物成瘾的药物是可用的，但 这些药物受到副作用、严格的处方指南和有限的可用性的限制。的 阿片类药物滥用的持续增加迫切需要新的治疗方法，提供持久，安全和成本- 有效干预过量和复发，但避免与当前成瘾相关的副作用 药物治疗使用药物特异性抗体的免疫药物疗法（即，免疫拮抗剂）阻断 阿片类药物活性和阻止靶向阿片类药物进入脑内是很有前途的治疗方法 副作用比传统的阿片类药物替代治疗更少。在这个SBIR项目中， 我们建议开发一类新的免疫治疗剂-纳米抗体（例如，单结构域 抗体，sdAbs），对海洛因、芬太尼具有高特异性和亲和力，用于治疗“杀手” 阿片类药物过量和滥用。我们希望阿片特异性纳米抗体会更好， 免疫治疗比传统的免疫球蛋白G为基础的药物抗体，因为纳米抗体是 非常坚固，高度抗变性，具有上级组织渗透性，并具有交叉能力 血脑屏障我们的目标是获得持久和高滴度的阿片类药物滥用抗体。 第一阶段的项目将集中在体外方法，用于产生特定的纳米抗体， 与噬菌体展示纳米抗体库中的“杀手阿片类药物”的亲和力。在第一阶段，我们将 阿片类药物的衍生物，使得它们可以在磁性载体上以高密度化学缀合， 珠，并分离出对海洛因和芬太尼特异性的高亲和力纳米抗体。此外，我们将设计 并表达单个单结构域抗体以产生二价（多价）纳米抗体或多价纳米抗体。 作为靶向两种或更多种阿片样物质的新治疗方式的纳米抗体缀合的纳米颗粒 同步这些纳米抗体将通过以下方式改进当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。一期工程的顺利完成将产生新的 阿片样物质特异性纳米抗体，并提供了一个框架的免疫治疗剂的发展， 其他滥用药物。在未来的第二阶段，我们将进行临床前体外和体内啮齿动物 单个纳米抗体和多价纳米抗体用于治疗性中和 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.