Background & Aims

Neuropathic pain is defined as a chronic disease caused by neurological injury or disease in peripheral and central nervous systems. Current analgesics are often inadequate due to the lack of efficacy and dose-limiting side effects [1]. The voltage-gated sodium channel Nav1.7 is primarily expressed in peripheral sensory neurons. Humans that lack the function of this channel are completely insensitive to pain [2]. Furthermore, several Nav1.7 inhibitors showed analgesic effect on neuropathic pain and erythromelalgia in clinical trials [3,4]. Nav1.7 inhibitor could therefore be a promising analgesic for a broad range of pain conditions. However, the development of Nav1.7 inhibitors has not yet been successfully progressed because small molecules are limited to have high selectivity due to high sequence similarity among Nav channel subtypes. Here, we present monoclonal antibodies which bind Nav1.7-specific extracellular domain and reduce pain behaviors and neuronal activity in spinal cord.

Methods

Mice were immunized with the Nav1.7 extracellular peptide-KLH conjugate. Monoclonal antibodies were established by hybridoma method and then humanized. Prepared antibodies (Antibody1 and 2) were evaluated about binding affinity to immunogen peptide by competitive ELISA and binding to Nav1.7 expressed in HEK cells by immunocytochemistry. Inhibitory activity of the antibodies on Nav1.7 sodium channel was measured using Nav1.7-expressing HEK cells and rat DRG neurons by manual patch clamp technique. The antibodies were injected intravenously in partial sciatic nerve ligation (pSNL) model rats to evaluate analgesic effect using von Frey filaments (n=8 to 10). Lastly, neuronal activity in dorsal horn of spinal cord in pSNL rats (n=6) was measured by in vivo electrophysiology after antibody1 i.v. injection (0.5, 5, and 15mg/kg) with/without stimulation on hindlimb by von Frey filaments. The effect of motor function by i.v. treatment of antibody1 (15mg/kg) was evaluated by rota-rod test.

Results

Prepared monoclonal antibodies had high affinity to immunogen peptide (IC50, antibody1: 1.52nM, antibody2: 0.73nM) with high selectivity among Nav channel subtypes (IC50, antibody 1 and 2: >1000nM) and had ability to bind Nav1.7 chanels expressed on HEK cells. The antibodies significantly inhibited current of Nav1.7 sodium channels expressed in HEK cells and firing in rat DRG neurons compared to negative control antibody. Next, intravenous injection of each antibody to pSNL rats showed the potent analgesic effect which lasted at least 96 hours. Lastly, neuronal firing in spinal cord was dose-dependently reduced by antibody1 treatment with/without stimulation on footpad. The efficacy on pain behavior and reduction of firing in spinal cord by treatment of antibody1 were highly correlated. The antibody1 did not affect motor function in rota-rod test.

Conclusions

These results suggest that our novel antibodies against Nav1.7 sodium channel would be promising analgesic drugs.

References

[1] Thouaye M. & Yalcin I. (2023). Neuropathic pain: From actual pharmacological treatments to new therapeutic horizons. Pharmacol Ther., 251, 108546.
[2] Dib-Hajj SD. et al. (2017). Sodium channels in pain disorders: pathophysiology and prospects for treatment. Pain. 158, S97-S107.
[3] Faber CG. et al. (2023). Efficacy and safety of vixotrigine in idiopathic or diabetes-associated painful small fibre neuropathy (CONVEY): a phase 2 placebo-controlled enriched-enrolment randomised withdrawal study. EClinicalMedicine. 59, 101971.
[4] Goldberg YP. et al. et al. (2012). Treatment of Na(v)1.7-mediated pain in inherited erythromelalgia using a novel sodium channel blocker. Pain. 153(1), 80-85.

Presenting Author

Sosuke Yoneda

Poster Authors

Sosuke Yoneda

PhD

Shionogi & Co., Ltd.

Lead Author

Daisuke Uta

University of Toyama

Lead Author

Kana Yasufuku

Shionogi & Co., Ltd.

Lead Author

Takuya Yamane

Shionogi & Co., Ltd.

Lead Author

Saho Yoshioka

Shionogi & Co., Ltd.

Lead Author

Keiko Takasu

Shionogi & Co., Ltd.

Lead Author

Takaya Izumi

Shionogi & Co., Ltd.

Lead Author

Sayaka Fujita

Shionogi & Co., Ltd.

Lead Author

Daiki Nakamori

Shionogi & Co., Ltd.

Lead Author

Shiori Kawasaki

Shionogi & Co., Ltd.

Lead Author

Tatsuya Takahashi

Shionogi & Co., Ltd.

Lead Author

Mai Yoshikawa

Shionogi & Co., Ltd.

Lead Author

Koichi Ogawa

Shionogi & Co., Ltd.

Lead Author

Erika Kasai

Shionogi & Co., Ltd.

Lead Author

Topics

  • Treatment/Management: Pharmacology: Monoclonal Antibodies, Biologics, and Biosimilars