Drug Design and Synthesis Section, Molecular Targets and Medications Discovery Branch, Intramural Research Program, National Institute on Drug Abuse and the National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Department of Health and Human Services, Bethesda, Maryland, USA.
Laboratory of Adjuvant and Antigen Research, U.S. Military HIV Research Program, Walter Reed Army Institute of Research, Silver Spring, Maryland, USA.
J Labelled Comp Radiopharm. 2020 Nov;63(13):564-571. doi: 10.1002/jlcr.3880. Epub 2020 Sep 28.
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-[ H ]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-[ H ]methyl]-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)-3-mercaptopropanamide.
设计并合成了一种氘代半抗原，这对半抗原-蛋白质缀合物的残留半抗原和稳定性的未来研究至关重要。这种半抗原，6-AmHap，被选为一种海洛因疫苗的候选物，目前正在进行 I 期临床试验。我们成功地将马来酰亚胺-巯基生物偶联策略应用于我们的海洛因疫苗中，通过三苯甲基保护的 3-巯基丙酰胺连接子将半抗原 6-AmHap 与免疫原性载体蛋白（破伤风类毒素，TT）连接起来。该疫苗诱导的抗体已被发现对几种阿片类药物具有活性，包括海洛因及其代谢物，重要的是，不会影响到其他替代疼痛治疗药物，如美沙酮。据我们所知，还没有其他海洛因疫苗的半抗原被氘代，但这个工具可能在研究产品释放过程中的残留半抗原和半抗原-蛋白质缀合物的长期稳定性计划方面非常重要，这是 FDA 监管要求的一部分。氢可酮是合成氘代 6-AmHap 的起始原料，C6 处有一个稳定的酰胺，C3 处有一个 3-巯基丙酰胺连接子。所需的氘代产物制备为二硫化物，3,3'-二磺酰基双（N-（（7S,7aR,12bS）-7-乙酰氨基-3-[ H ]甲基）-2,3,4,4a,5,6,7,7a-八氢-1H-4,12-甲烷苯并呋喃[3,2-e]异喹啉-9-基）丙酰胺），可轻松还原形成所需的半抗原，N-（（4aR,7S,7aR,12bS）-7-乙酰氨基-3-[ H ]甲基）-2,3,4,4a,5,6,7,7a-八氢-1H-4,12-甲烷苯并呋喃[3,2-e]异喹啉-9-基）-3-巯基丙酰胺。
