Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
Mol Pharm. 2012 Feb 6;9(2):281-9. doi: 10.1021/mp200376c. Epub 2011 Dec 23.
Ghrelin, an enteric peptide hormone linked to the pathophysiology of obesity has been a therapeutic target of great interest over the past decade. Many research efforts have focused on the antagonism of ghrelin's endogenous receptor GHSR1a, which is found along ascending vagal afferent fibers, as well as in the arcuate nucleus of the hypothalamus. Additionally, peptidic inhibitors of ghrelin O-acyltransferase, the enzyme responsible for the paracrine activation of ghrelin, have recently been studied. Our research has taken an alternative immunological approach, studying both active and passive vaccination as a means to sequester ghrelin in the periphery, with the original discovery in rat of decreased feed efficiency and adiposity, as well as increased metabolic activity. Using our previous hapten designs as a stepping-stone, three monoclonal antibodies (JG2, JG3, and JG4) were procured against ghrelin and tested in vivo. While mAb JG4 had the highest affinity for ghrelin, it failed to attenuate the orexigenic effects of food deprivation on energy metabolism or food intake in mice. However, animals that were administered a combination of JG3:JG4 (termed a doublet) or JG2:JG3:JG4 (termed a triplet) demonstrated higher heat dispersion and rate of respiration (higher CO(2) emission and O(2) consumption) during a 24 h fast refeed. Mice administered the triplet cocktail of JG2:JG3:JG4 also demonstrated decreased food intake upon refeeding as compared to control animals. Recently, Lu and colleagues reported that a passive approach using a single, high affinity N-terminally directed monoclonal antibody did not abrogate the effects of endogenous ghrelin. Our current report corroborates this finding, yet, refutes that a monoclonal antibody approach cannot be efficacious. Rather, we find that a multiple monoclonal antibody (oligoclonal) approach can reproduce the underlying logic to previously reported efficacies using active vaccinations.
胃饥饿素是一种与肥胖的病理生理学相关的肠肽激素,在过去十年中一直是一个非常有吸引力的治疗靶点。许多研究都集中在拮抗胃饥饿素的内源性受体 GHSR1a 上,该受体存在于上升的迷走传入纤维中,也存在于下丘脑弓状核中。此外,最近还研究了胃饥饿素 O-酰基转移酶的肽类抑制剂,该酶负责胃饥饿素的旁分泌激活。我们的研究采用了一种替代的免疫学方法,研究主动和被动免疫接种作为在外周隔离胃饥饿素的一种手段,最初在大鼠中发现了摄食量减少和肥胖,以及代谢活性增加。我们利用之前的半抗原设计作为垫脚石,获得了针对胃饥饿素的三种单克隆抗体(JG2、JG3 和 JG4),并在体内进行了测试。虽然 mAb JG4 对胃饥饿素具有最高的亲和力,但它未能减弱食物剥夺对能量代谢或食物摄入的食欲刺激作用。然而,给予 JG3:JG4(称为二联体)或 JG2:JG3:JG4(称为三联体)组合的动物在 24 小时禁食后再喂养时表现出更高的热量散发和呼吸率(更高的 CO2 排放和 O2 消耗)。给予 JG2:JG3:JG4 三联体鸡尾酒的小鼠在再喂养时的食物摄入量也比对照动物减少。最近,Lu 及其同事报道,使用单一的、高亲和力的 N 端定向单克隆抗体的被动方法并不能消除内源性胃饥饿素的作用。我们目前的报告证实了这一发现,但反驳了单克隆抗体方法不能有效的观点。相反,我们发现,使用多克隆抗体(多克隆)方法可以复制之前使用主动免疫接种报告的疗效的基本逻辑。