Jalah Rashmi, Torres Oscar B, Mayorov Alexander V, Li Fuying, Antoline Joshua F G, Jacobson Arthur E, Rice Kenner C, Deschamps Jeffrey R, Beck Zoltan, Alving Carl R, Matyas Gary R
†Laboratory of Adjuvant and Antigen Research, US Military HIV Research Program, Walter Reed Army Institute of Research, 503 Robert Grant Avenue, Silver Spring, Maryland 20910, United States.
‡U.S. Military HIV Research Program, Henry M. Jackson Foundation for the Advancement of Military Medicine, 6720A Rockledge Drive, Bethesda, Maryland 20817, United States.
Bioconjug Chem. 2015 Jun 17;26(6):1041-53. doi: 10.1021/acs.bioconjchem.5b00085. Epub 2015 Jun 5.
Vaccines against drugs of abuse have induced antibodies in animals that blocked the biological effects of the drug by sequestering the drug in the blood and preventing it from crossing the blood-brain barrier. Drugs of abuse are too small to induce antibodies and, therefore, require conjugation of drug hapten analogs to a carrier protein. The efficacy of these conjugate vaccines depends on several factors including hapten design, coupling strategy, hapten density, carrier protein selection, and vaccine adjuvant. Previously, we have shown that 1 (MorHap), a heroin/morphine hapten, conjugated to tetanus toxoid (TT) and mixed with liposomes containing monophosphoryl lipid A [L(MPLA)] as adjuvant, partially blocked the antinociceptive effects of heroin in mice. Herein, we extended those findings, demonstrating greatly improved vaccine induced antinociceptive effects up to 3% mean maximal potential effect (%MPE). This was obtained by evaluating the effects of vaccine efficacy of hapten 1 vaccine conjugates with varying hapten densities using two different commonly used carrier proteins, TT and cross-reactive material 197 (CRM197). Immunization of mice with these conjugates mixed with L(MPLA) induced very high anti-1 IgG peak levels of 400-1500 μg/mL that bound to both heroin and its metabolites, 6-acetylmorphine and morphine. Except for the lowest hapten density for each carrier, the antibody titers and affinity were independent of hapten density. The TT carrier based vaccines induced long-lived inhibition of heroin-induced antinociception that correlated with increasing hapten density. The best formulation contained TT with the highest hapten density of ≥30 haptens/TT molecule and induced %MPE of approximately 3% after heroin challenge. In contrast, the best formulation using CRM197 was with intermediate 1 densities (10-15 haptens/CRM197 molecule), but the %MPE was approximately 13%. In addition, the chemical synthesis of 1, the optimization of the conjugation method, and the methods for the accurate quantification of hapten density are described.
针对滥用药物的疫苗已在动物体内诱导产生抗体，这些抗体通过将药物隔离在血液中并阻止其穿过血脑屏障来阻断药物的生物学效应。滥用药物分子太小，无法诱导抗体产生，因此需要将药物半抗原类似物与载体蛋白结合。这些结合疫苗的疗效取决于几个因素，包括半抗原设计、偶联策略、半抗原密度、载体蛋白选择和疫苗佐剂。此前，我们已经表明，与破伤风类毒素（TT）偶联并与含有单磷酰脂质A的脂质体[L（MPLA）]作为佐剂混合的海洛因/吗啡半抗原1（MorHap），部分阻断了海洛因对小鼠的镇痛作用。在此，我们扩展了这些发现，证明疫苗诱导的镇痛作用有了显著改善，平均最大潜在效应（%MPE）高达3%。这是通过使用两种不同的常用载体蛋白TT和交叉反应物质197（CRM197）评估不同半抗原密度的半抗原1疫苗偶联物的疫苗疗效来实现的。用这些与L（MPLA）混合的偶联物免疫小鼠，诱导出400 - 1500μg/mL的非常高的抗-1 IgG峰值水平，该抗体与海洛因及其代谢物6-乙酰吗啡和吗啡都能结合。除了每种载体的最低半抗原密度外，抗体滴度和亲和力与半抗原密度无关。基于TT载体的疫苗诱导了对海洛因诱导的镇痛作用的长期抑制，这与半抗原密度的增加相关。最佳配方包含半抗原密度≥30个半抗原/TT分子的TT，在海洛因激发后诱导的%MPE约为3%。相比之下，使用CRM197的最佳配方是中等密度的1（10 - 15个半抗原/CRM197分子），但%MPE约为13%。此外，还描述了1的化学合成、偶联方法的优化以及半抗原密度的准确定量方法。
