Novel Vaccine That Blunts Fentanyl Effects and Sequesters Ultrapotent Fentanyl Analogues
Novel Vaccine That Blunts Fentanyl Effects and Sequesters Ultrapotent Fentanyl Analogues
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DOI：
10.1021/acs.molpharmaceut.0c00497
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发表时间：
2020-09-08
影响因子：
4.9
通讯作者：
Matyas, Gary R.
中科院分区：
文献类型：
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作者：
Barrientos, Rodell C.;Bow, Eric W.;Matyas, Gary R.
主动免疫是一种新兴的潜在模式，可在阿片类药物流行病中对抗致命的过量用药。在这项研究中，我们描述了一种有效的芬太尼疫苗的设计，合成，配方和动物试验。该疫苗制剂由与破伤风类毒素（TT）缀合的新型芬太尼半抗原组成，并用含有吸附在氢氧化铝上的单磷酰脂质A的脂质体作为佐剂。接头和半抗原N-苯基- N-（1-（4-（3-（三苯甲基硫基）丙酰胺基）苯乙基）哌啶-4-基）丙酰胺分别使用胺-N-羟基琥珀酰亚胺-酯和硫醇-马来酰亚胺反应化学顺序缀合至TT。缀合是容易的、有效的和可再现的，其中蛋白质回收率>98%并且半抗原密度为每个载体蛋白分子30-35个。在小鼠中，免疫诱导针对半抗原的高且稳健的抗体终点滴度，其数量级>10（6）。抗血清在体外结合芬太尼、卡芬太尼、环丙基芬太尼、对氟芬太尼和呋喃基芬太尼，抗体-药物解离常数在0.36-4.66 nM范围内。未观察到与纳洛酮、纳洛酮、美沙酮或丁丙诺啡的交叉反应性。在体内，免疫将芬太尼的抗伤害性剂量-反应曲线转移到更高的剂量。总的来说，这些临床前结果展示了针对芬太尼的潜在疫苗的所需特性，并证明了免疫对抗芬太尼诱导效应的可行性。
Active immunization is an emerging potential modality to combat fatal overdose amid the opioid epidemic. In this study, we described the design, synthesis, formulation, and animal testing of an efficacious vaccine against fentanyl. The vaccine formulation is composed of a novel fentanyl hapten conjugated to tetanus toxoid (TT) and adjuvanted with liposomes containing monophosphoryl lipid A adsorbed on aluminum hydroxide. The linker and hapten N-phenyl- N-(1-(4-(3-(tritylthio) propanamido)phenethyl)piperidin-4-yl) propionamide were conjugated sequentially to TT using amine-N-hydroxysuccinimide-ester and thiol-maleimide reaction chemistries, respectively. Conjugation was facile, efficient, and reproducible with a protein recovery of >98% and a hapten density of 30-35 per carrier protein molecule. In mice, immunization induced high and robust antibody endpoint titers in the order of >10(6) against the hapten. The antisera bound fentanyl, carfentanil, cyclopropyl fentanyl, para-fluorofentanyl, and furanyl fentanyl in vitro with antibody-drug dissociation constants in the range of 0.36-4.66 nM. No cross-reactivity to naloxone, naltrexone, methadone, or buprenorphine was observed. In vivo, immunization shifted the antinociceptive dose-response curve of fentanyl to higher doses. Collectively, these preclinical results showcased the desired traits of a potential vaccine against fentanyl and demonstrated the feasibility of immunization to combat fentanyl-induced effects.