A deuterated hapten was designed and synthesized that will be essential for a future study of residual hapten and stability of a hapten-protein conjugate. This hapten, 6-AmHap, was chosen for a heroin vaccine that is now slated for a Phase 1 clinical trial. A maleimide-thiol bioconjugation strategy was successfully applied to our heroin vaccine to connect the hapten 6-AmHap with an immunogenic carrier protein (tetanus toxoid, TT) through a trityl-protected 3-mercaptopropanamide linker. The antibodies induced by the vaccine have been found to have activity against several opioids, including heroin and its metabolites, and, importantly, leave alternate pain treatment medications such as methadone untouched. To the best of our knowledge, no other hapten for a heroin vaccine has been deuterated, yet this tool may prove to be of great importance in the study of residual hapten during product release and the long-term stability program of a hapten-protein conjugate as part of FDA regulatory requirements. Hydrocodone was the starting material for the synthesis of the deuterated 6-AmHap, with a stable amide at C6 and a 3-mercaptopropanamide linker attached at C3. The desired deuterated product was prepared as the disulfide, 3,3'-disulfanediylbis(N-((7S,7aR,12bS)-7-acetamido-3-[2 H3 ]methyl)-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)propanamide), that could be easily reduced to form the needed hapten, N-((4aR,7S,7aR,12bS)-7-acetamido-3-[2 H3 ]methyl]-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)-3-mercaptopropanamide.

中文翻译：

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用于测定海洛因疫苗药物产品中半抗原密度的氘代 6-AmHap 内标的合成

设计和合成了一种氘代半抗原，这对于未来研究残留半抗原和半抗原-蛋白质缀合物的稳定性至关重要。这种半抗原 6-AmHap 被选择用于海洛因疫苗，该疫苗现在计划进行 1 期临床试验。马来酰亚胺-硫醇生物偶联策略已成功应用于我们的海洛因疫苗，通过三苯甲基保护的 3-巯基丙酰胺接头将半抗原 6-AmHap 与免疫原性载体蛋白（破伤风类毒素，TT）连接起来。已发现由疫苗诱导的抗体对包括海洛因及其代谢物在内的几种阿片类药物具有活性，并且重要的是，不会触及美沙酮等替代疼痛治疗药物。据我们所知，海洛因疫苗没有其他半抗原被氘化，然而，该工具可能被证明在产品发布期间残留半抗原的研究以及作为 FDA 监管要求的一部分的半抗原-蛋白质偶联物的长期稳定性计划中具有重要意义。氢可酮是合成氘代 6-AmHap 的起始材料，在 C6 处具有稳定的酰胺，在 C3 处连接有 3-巯基丙酰胺接头。所需的氘代产物制备为二硫化物 3,3'-二硫烷二基双(N-((7S,7aR,12bS)-7-acetamido-3-[2 H3]methyl)-2,3,4,4a,5 ,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)propanamide)，可以很容易地还原形成所需的半抗原，N-((4aR,7S ,7aR,12bS)-7-acetamido-3-[2 H3 ]methyl]-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3,2-e ]isoquinolin-9-yl)-3-mercaptopropanamide。