Korean J Pain.  2022 Apr;35(2):173-182. 10.3344/kjp.2022.35.2.173.

Anti-nociceptive effects of dual neuropeptide antagonist therapy in mouse model of neuropathic and inflammatory pain

• Kim MS
• Kim BY
• Saghetlians A
• Zhang X
• Okida T
• Kim SY

• KMID: 2527768
• DOI: http://doi.org/10.3344/kjp.2022.35.2.173

Abstract

Background
Neurokinin-1 (NK1) and calcitonin gene-related peptide (CGRP) play a vital role in pain pathogenesis, and these proteins’ antagonists have attracted attention as promising pharmaceutical candidates. The authors investigated the antinociceptive effect of co-administration of the CGRP antagonist and an NK1 antagonist on pain models compared to conventional single regimens.
Methods
C57Bl/6J mice underwent sciatic nerve ligation for the neuropathic pain model and were injected with 4% formalin into the hind paw for the inflammatory pain model. Each model was divided into four groups: vehicle, NK1 antagonist, CGRP antagonist, and combination treatment groups. The NK1 antagonist aprepitant (BIBN4096, 1 mg/kg) or the CGRP antagonist olcegepant (MK-0869, 10 mg/ kg) was injected intraperitoneally. Mechanical allodynia, thermal hypersensitivity, and anxiety-related behaviors were assessed using the von Frey, hot plate, and elevated plus-maze tests. The flinching and licking responses were also evaluated after formalin injection.
Results
Co-administration of aprepitant and olcegepant more significantly alleviated pain behaviors than administration of single agents or vehicle, increasing the mechanical threshold and improving the response latency. Anxiety-related behaviors were also markedly improved after dual treatment compared with either naive mice or the neuropathic pain model in the dual treatment group. Flinching frequency and licking response after formalin injection decreased significantly in the dual treatment group. Isobolographic analysis showed a meaningful additive effect between the two compounds.
Conclusions
A combination pharmacological therapy comprised of multiple neuropeptide antagonists could be a more effective therapeutic strategy for alleviating neuropathic or inflammatory pain.

Keyword

Analgesics; Anxiety; Calcitonin Gene-Related Peptide Receptor Antagonists; Hyperalgesia; Neuralgia; Neurokinin-1 Receptor Antagonists; Neuropeptides; Nociception; Pain

Figure

• 

  Fig. 1 Change of mechanical threshold measured with von Frey test after administrating aprepitant (APR) and olcegepant (OLC). (A) The mechanical threshold in naive mice increased significantly with co-administration of both drugs intraperitoneally. (B) More remarkable improvement of mechanical threshold was observed in the dual-treated neuropathic pain model compared to single-treated groups or vehicle. The error bars indicate standard deviation. *P < 0.05 compared with vehicle.

• 

  Fig. 2 Effects on the thermal hypersensitivity. (A) Combination treatment by aprepitant (APR) and olcegepant (OLC) significantly attenuated the thermal hypersensitivity over time in naive mice. (B) There was a significant increase in response latency for thermal hypersensitivity in the neuropathic pain model in dual treatment groups. There were no changes in response latency in either a single treatment group or vehicle. The error bars indicate standard deviation. *P < 0.05 compared with vehicle.

• 

  Fig. 3 Effects on formalin-induced inflammatory pain. (A) The number of flinching significantly decreased in the dual treatment group from 30 minutes after formalin injection compared to either single or vehicle administration. (B) Licking cumulative time in the dual treatment group was significantly shorter than in other groups in phase II. APR: aprepitant, OLC: olcegepant. The error bars indicate standard deviation. *P < 0.05 compared with vehicle.

• 

  Fig. 4 Effect of co-administration on anxiety-like behavior measured by the elevated plus-maze test. (A) In both naive mice and neuropathic pain models, the co-administration of aprepitant (APR) and olcegepant (OLC) markedly increased the time spent in the open arms compared with the mice treated by single compound or vehicle. (B) In the dual treatment group, naive mice or neuropathic pain model significantly spent less time than the single compound or vehicle treated mice in the close arms. (C) The total arm entries didn’t show significant differences among groups, either naive mice or neuropathic pain model. The error bars indicate standard deviation. *P < 0.05 compared with vehicle or single compound groups.

• 

  Fig. 5 Dose-response effects of the compounds analyzed by the flinching number after formalin injection. Data are presented as the percent of pain inhibition effect (% PIE). (A) Olcegepant (OLC). (B) Aprepitant (APR). (C) Combination treatment. The error bars indicate standard deviation. *P < 0.05 compared with vehicle.

• 

  Fig. 6 Isobologram for the interaction of olcegepant and aprepitant. The diagonal line connecting two 50% effective dose (ED50) points is the theoretical line of additivity. The experimental ED50 point was decreased significantly below the line of additivity, indicating a meaningful additive effect.

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