Lay Line Genomics, SpA, Roma, Italy.
PLoS One. 2012;7(3):e32212. doi: 10.1371/journal.pone.0032212. Epub 2012 Mar 5.
Most forms of chronic pain are inadequately treated by present therapeutic options. Compelling evidence has accumulated, demonstrating that Nerve Growth Factor (NGF) is a key modulator of inflammatory and nociceptive responses, and is a promising target for the treatment of human pathologies linked to chronic and inflammatory pain. There is therefore a growing interest in the development of therapeutic molecules antagonising the NGF pathway and its nociceptor sensitization actions, among which function-blocking anti-NGF antibodies are particularly relevant candidates.In this respect, the rat anti-NGF αD11 monoclonal antibody (mAb) is a potent antagonist, able to effectively antagonize rodent and human NGF in a variety of in vitro and in vivo systems. Here we show that mAb αD11 displays a significant analgesic effect in two different models of persistent pain in mice, with a remarkable long-lasting activity. In order to advance αD11 mAb towards its clinical application in man, anti-NGF αD11 mAb was humanized by applying a novel single cycle strategy based on the a priori experimental determination of the crystal and molecular structure of the parental Fragment antigen-binding (Fab). The humanized antibody (hum-αD11) was tested in vitro and in vivo, showing that the binding mode and the NGF neutralizing biological activities of the parental antibody are fully preserved, with even a significant affinity improvement. The results firmly establish hum-αD11 as a lead candidate for clinical applications in a therapeutic area with a severe unmet medical need. More generally, the single-cycle structure-based humanization method represents a considerable improvement over the standard humanization methods, which are intrinsically empirical and require several refinement cycles.
目前的治疗选择无法充分治疗大多数形式的慢性疼痛。大量确凿的证据表明，神经生长因子（NGF）是炎症和伤害性反应的关键调节剂，是治疗与慢性和炎症性疼痛相关的人类病理的有前途的靶点。因此，人们越来越关注开发拮抗肿瘤坏死因子途径及其伤害感受器敏化作用的治疗分子，其中阻断 NGF 通路的功能阻断性抗 NGF 抗体是特别相关的候选物。
在这方面，大鼠抗 NGF αD11 单克隆抗体（mAb）是一种有效的拮抗剂，能够在多种体外和体内系统中有效拮抗啮齿动物和人 NGF。在这里，我们显示 mAb αD11 在两种不同的小鼠持续性疼痛模型中显示出显著的镇痛作用，具有显著的长效活性。为了将 mAb αD11 推进到其在人类中的临床应用，通过应用基于母体片段抗原结合（Fab）晶体和分子结构的先验实验确定的新型单循环策略对抗 NGF αD11 mAb 进行了人源化。对人源化抗体（hum-αD11）进行了体外和体内测试，结果表明，母体抗体的结合模式和中和 NGF 的生物学活性得到了完全保留，甚至亲和力也得到了显著提高。这些结果牢固确立了 hum-αD11 作为在一个严重未满足医疗需求的治疗领域进行临床应用的候选药物。更普遍地说，基于单循环结构的人源化方法代表了对标准人源化方法的重大改进，标准人源化方法本质上是经验性的，需要几个改进循环。
