Department of Biomedical Science, University of Sheffield, Sheffield, UK.
Department of Pharmacology, School of Pharmacy, University of Reading, Whiteknights Campus, Reading, UK.
Life Sci Alliance. 2023 Apr 11;6(6). doi: 10.26508/lsa.202201631. Print 2023 Jun.
Chronic pain affects one in five people across human societies, with few therapeutic options available. Botulinum neurotoxin (BoNT) can provide long-lasting pain relief by inhibiting local release of neuropeptides and neurotransmitters, but its highly paralytic nature has limited its analgesic potential. Recent advances in protein engineering have raised the possibility of synthesising non-paralysing botulinum molecules for translation to pain sufferers. However, the synthesis of these molecules, via several synthetic steps, has been challenging. Here, we describe a simple platform for safe production of botulinum molecules for treating nerve injury-induced pain. We produced two versions of isopeptide-bonded BoNT from separate botulinum parts using an isopeptide bonding system. Although both molecules cleaved their natural substrate, SNAP25, in sensory neurons, the structurally elongated iBoNT did not cause motor deficit in rats. We show that the non-paralytic elongated iBoNT targets specific cutaneous nerve fibres and provides sustained pain relief in a rat nerve injury model. Our results demonstrate that novel botulinum molecules can be produced in a simple and safe manner and be useful for treating neuropathic pain.
慢性疼痛影响着全球五分之一的人群，然而目前可供选择的治疗方法却寥寥无几。肉毒杆菌神经毒素（BoNT）通过抑制神经肽和神经递质的局部释放，可以提供持久的疼痛缓解，但由于其高度致瘫的特性，限制了其在镇痛方面的潜力。近年来，蛋白质工程的进步提出了合成非致瘫肉毒杆菌分子以用于疼痛患者的可能性。然而，通过多个合成步骤来合成这些分子具有一定的挑战性。在这里，我们描述了一个简单的平台，用于安全生产用于治疗神经损伤引起的疼痛的肉毒杆菌分子。我们使用异肽键合系统，从两个单独的肉毒杆菌部分生产了两种异肽键合的 BoNT。尽管这两种分子都能在感觉神经元中切割其天然底物 SNAP25，但结构上拉长的 iBoNT 不会在大鼠中引起运动缺陷。我们表明，非致瘫的拉长 iBoNT 靶向特定的皮肤神经纤维，并在大鼠神经损伤模型中提供持续的疼痛缓解。我们的结果表明，可以以简单和安全的方式生产新型肉毒杆菌分子，并且对治疗神经病理性疼痛有用。
