Department of Pharmacology, College of Medicine, The University of Arizona, Tucson, AZ, United States.
Department of Molecular Pathobiology, College of Dentistry, New York University, New York, NY, United States.
Pain. 2023 Jul 1;164(7):1473-1488. doi: 10.1097/j.pain.0000000000002850. Epub 2022 Dec 19.
Vascular endothelial growth factor A (VEGF-A) is a pronociceptive factor that causes neuronal sensitization and pain. We reported that blocking the interaction between the membrane receptor neuropilin 1 (NRP1) and VEGF-A-blocked VEGF-A-mediated sensory neuron hyperexcitability and reduced mechanical hypersensitivity in a rodent chronic neuropathic pain model. These findings identified the NRP1-VEGF-A signaling axis for therapeutic targeting of chronic pain. In an in-silico screening of approximately 480 K small molecules binding to the extracellular b1b2 pocket of NRP1, we identified 9 chemical series, with 6 compounds disrupting VEGF-A binding to NRP1. The small molecule with greatest efficacy, 4'-methyl-2'-morpholino-2-(phenylamino)-[4,5'-bipyrimidin]-6(1H)-one, designated NRP1-4, was selected for further evaluation. In cultured primary sensory neurons, VEGF-A enhanced excitability and decreased firing threshold, which was blocked by NRP1-4. In addition, NaV1.7 and CaV2.2 currents and membrane expression were potentiated by treatment with VEGF-A, and this potentiation was blocked by NRP1-4 cotreatment. Neuropilin 1-4 reduced VEGF-A-mediated increases in the frequency and amplitude of spontaneous excitatory postsynaptic currents in dorsal horn of the spinal cord. Neuropilin 1-4 did not bind to more than 300 G-protein-coupled receptors and receptors including human opioids receptors, indicating a favorable safety profile. In rats with spared nerve injury-induced neuropathic pain, intrathecal administration of NRP1-4 significantly attenuated mechanical allodynia. Intravenous treatment with NRP1-4 reversed both mechanical allodynia and thermal hyperalgesia in rats with L5/L6 spinal nerve ligation-induced neuropathic pain. Collectively, our findings show that NRP1-4 is a first-in-class compound targeting the NRP1-VEGF-A signaling axis to control voltage-gated ion channel function, neuronal excitability, and synaptic activity that curb chronic pain.
血管内皮生长因子 A（VEGF-A）是一种致痛因子，可导致神经元敏化和疼痛。我们曾报道，阻断膜受体神经纤毛蛋白 1（NRP1）与 VEGF-A 之间的相互作用，可阻断 VEGF-A 介导的感觉神经元过度兴奋，并减轻啮齿动物慢性神经病理性疼痛模型中的机械性超敏反应。这些发现确定了 NRP1-VEGF-A 信号轴可作为治疗慢性疼痛的靶点。在针对 NRP1 细胞外 b1b2 口袋的约 480K 小分子的计算机筛选中，我们鉴定出 9 个化学系列，其中 6 个化合物可破坏 VEGF-A 与 NRP1 的结合。效力最大的小分子 4′-甲基-2′-吗啉-2-（苯氨基）-[4,5′-联嘧啶]-6(1H)-酮，命名为 NRP1-4，被选为进一步评估的对象。在培养的原代感觉神经元中，VEGF-A 增强了兴奋性并降低了放电阈值，而 NRP1-4 可阻断这一作用。此外，VEGF-A 处理可增强 NaV1.7 和 CaV2.2 电流和膜表达，而 NRP1-4 共同处理可阻断这种增强作用。NRP1-4 减少了 VEGF-A 介导的脊髓背角中自发性兴奋性突触后电流频率和幅度的增加。NRP1-4 不与 300 多种 G 蛋白偶联受体和受体（包括人类阿片受体）结合，表明其具有良好的安全性。在 spared nerve injury 诱导的神经病理性疼痛大鼠中，鞘内给予 NRP1-4 可显著减轻机械性痛觉过敏。静脉给予 NRP1-4 可逆转 L5/L6 脊神经结扎诱导的神经病理性疼痛大鼠的机械性痛觉过敏和热痛觉过敏。总之，我们的研究结果表明，NRP1-4 是一种新型化合物，可靶向 NRP1-VEGF-A 信号轴，控制电压门控离子通道功能、神经元兴奋性和突触活动，从而抑制慢性疼痛。
