T. brucei: next-generation platform for immunization against drugs of abuse

描述（由申请人提供）：处方阿片类药物（短效mopo -r激动剂，如羟考酮）的滥用和成瘾在美国已达到流行病的程度。阻止从滥用到成瘾的发展轨迹的治疗选择是有限的。因此，为了这种预防目的而出现了预防药物滥用的疫苗概念。然而，小分子如滥用药物不是高度免疫原性的，因此标准方法包括将抗原表位（例如，羟考酮）偶联到蛋白质免疫原上。假设有效的疫苗接种可以通过大量高亲和力抗体的作用，使滥用药物在血液中被隔离（从而失活）。然而，标准方法有许多技术限制，可能会限制它们的可靠性和有效性。因此，CEBRA提案的目标是开发一种机制新颖的疫苗接种平台，该平台基于细胞外原生动物寄生虫布氏锥虫（T. brucei）外壳中抗原表位的有序和高密度呈递。这种生物进化的抗原呈递机制导致强烈的b细胞反应和b细胞记忆，这对有效的疫苗接种至关重要。因此，本多学科建议的目标1将发展第一个。brucei小分子疫苗接种平台，利用氧可酮抗原片段的有序呈现来修饰在brucei上转基因表达的分选酶“标签”。然后用灭活的布氏体（没有潜在致病性）接种大鼠模型，并优化选择性抗氧可酮抗体的产生和时间过程。目标2将关注该方法的“技术证明”验证。这将通过优化大鼠抗羟考酮疫苗接种的有效性，通过药代动力学（例如，血液羟考酮水平降低）和药效学（阻断羟考酮诱导的镇痛和静脉注射羟考酮自我给药）参数进行监测。因此，这种针对滥用小分子（以羟考酮为原型）的尖端疫苗接种策略的发展，在临床转化和药物滥用研究方面具有相当大的未来潜力。

DESCRIPTION (provided by applicant): Abuse and addiction to prescription opioids (short-acting MOP-r agonists such as oxycodone) have reached epidemic proportions in the US. Therapeutic options to block the progression of trajectory from abuse to addiction are limited. The concept of vaccination against drugs of abuse has therefore emerged for such a preventive purpose. However small molecules such as drugs of abuse are not highly immunogenic, therefore standard approaches include conjugation of the antigen epitope (e.g., oxycodone) to a protein immunogen. Effective vaccination is postulated to result in sequestration (and thus inactivation) of the drug of abuse in the bloodstream, through the action of a sufficient population of high-affinity antibodies. However, standard approaches have a number of technical limitations that can limit their reliability and effectiveness. The goal of this CEBRA proposal is therefore to develop a mechanistically novel vaccination platform based on the orderly and high-density presentation of antigen epitopes in the coat of the extracellular protozoan parasite Trypanosoma brucei (T. brucei). This biologically-evolved antigen-presentation mechanism results in strong B-cell responses and B-cell memory, crucial to effective vaccination. Aim 1 of this multi- disciplinary proposal will therefore develop the first . brucei platform for small molecule vaccination, utilizing orderly presentation of oxycodone antigen moieties to decorate sortase "tags" transgenically expressed on the coat of T. brucei. Vaccination with inactivated T. brucei (having no potential for pathogenicity) will then be modeled in rats, and the production and time course of selective anti-oxycodone antibodies will be optimized. Aim 2 will then focus on the "proof-of-technology" validation of this approach. This will be done through optimization of the effectiveness of anti-oxycodone vaccination in rats, monitored by pharmacokinetic (e.g., decrease in blood oxycodone levels) and pharmacodynamic (blockade of oxycodone-induced analgesia and of i.v. oxycodone self-administration) parameters. The development of this cutting-edge vaccination strategy for abused small molecules (with oxycodone as a prototype) therefore has considerable future potential for clinical translation, and for drug abuse research.