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

描述(申请人提供)：处方类阿片类药物(如羟考酮等短效MOP-r激动剂)的滥用和成瘾在美国已经达到流行的程度。阻止从滥用到成瘾的轨迹进展的治疗选择有限。因此，为了这一预防目的，出现了针对滥用药物的疫苗接种的概念。然而，像滥用药物这样的小分子并不具有很强的免疫原性，因此标准的方法包括将抗原表位(例如羟考酮)连接到蛋白质免疫原上。有效的疫苗接种被认为是通过足够数量的高亲和力抗体的作用，导致血液中滥用药物的隔离(从而使其失活)。然而，标准方法有许多技术限制，可能会限制其可靠性和有效性。因此，CEBRA建议的目标是开发一种机械上新颖的疫苗接种平台，该平台基于细胞外原生动物布氏锥虫(T.brucei)外壳抗原表位的有序和高密度呈现。这种生物进化的抗原呈递机制导致了强大的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，这项多学科提案的目标1将发展第一个目标。布鲁西平台用于小分子疫苗接种，利用羟考酮抗原部分的有序呈现来装饰转基因表达在布鲁氏锥虫外衣上的索酸酶“标签”。然后，将在大鼠身上模拟灭活的布鲁氏旋毛虫(无致病潜力)疫苗，并优化选择性抗羟考酮抗体的产生和时间过程。然后，目标2将专注于该方法的“技术验证”验证。这将通过优化抗羟考酮疫苗在大鼠中的有效性来实现，通过药代动力学(例如，血羟考酮水平的降低)和药效学(阻断羟考酮诱导的镇痛和静脉注射)来监测。羟考酮自我给药)参数。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开发在临床翻译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.