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MOP-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，为防止滥用药物而进行疫苗接种的概念便应运而生。然而，小分子如滥用药物不是高度免疫原性的，因此标准方法包括抗原表位（例如，羟考酮）与蛋白质免疫原的结合。有效的疫苗接种被认为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（从而灭活）。然而，标准方法有一些技术限制，可能会限制其可靠性和有效性。因此，CEBRA提案的目标是开发一种基于细胞外原生动物寄生虫布氏锥虫（Trypanosoma brucei，T. brucei）。这种生物进化的抗原呈递机制导致强烈的B细胞应答和B细胞记忆，这对有效的疫苗接种至关重要。因此，这个多学科建议的目标1将发展第一个.布氏梭菌平台用于小分子疫苗接种，利用氧可酮抗原部分的有序呈递来修饰转基因表达在布氏梭菌外壳上的分选酶“标签”。布鲁塞。接种灭活T.然后在大鼠中建立布鲁氏菌（没有致病性的潜力）模型，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将侧重于这种方法的“技术证明”验证。这将通过优化大鼠中抗羟考酮疫苗接种的有效性来完成，通过药代动力学（例如，血液羟考酮水平降低）和药效学（羟考酮诱导的镇痛和静脉内羟考酮自我给药的阻断）参数。因此，这种针对滥用小分子的尖端疫苗接种策略（以羟考酮为原型）的开发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.