Korean J Pain.  2025 Apr;38(2):163-176. 10.3344/kjp.24301.

The analgesic and anti-inflammatory effects of a combined preparation based on the blunt-nosed viper’s venom and oregano essential oil

• Parsghyan LM
• Moghrovyan AV
• Poghosyan SSP
• Babajanyan MA
• Gaboyan MA
• Voskanyan AV
• Darbinyan AA

• KMID: 2566648
• DOI: http://doi.org/10.3344/kjp.24301

Abstract

Background
To relieve acute and inflammatory pain, preparations of plant and animal origin have been used. The present work aimed to study the analgesic and anti-inflammatory effectiveness of a combined preparation based on viper venom and essential oil. Determining effective routes of exposure, optimal doses, the duration of action of the preparation, and possible mechanisms of their action were the areas of interest.
Methods
Experiments were carried out on murine. Essential oil content was determined by gas chromatography– mass spectrometry equipment. The formalin, carrageenan, and hot plate tests were used. Certain methods for determining side effects were used as well. To determine the participation of cannabinoid and opioid receptors in the antinociceptive action of combined preparation, SR144528 and naloxone were used.
Results
The treatment of the ointment version of the preparation reduced inflammatory pain by more than 68% and decreased the volume of inflammatory edema by up to 36%. The involvement of cannabinoid receptors in the analgesic mechanism of the ointment was approximately 73%, and, for the opioid receptors, about 64%. Physiologically significant side effects were not observed.
Conclusions
The active components of the ointment are principally different in their mechanism of action and make it possible to relieve pain and inflammation both through the blockade of pain receptors of afferent nociceptive neurons (venom) as well as via cannabinoid and opioid receptors (essential oil).

Keyword

Acute Pain; Analgesics; Opioid; Cannabinoids; Drug Compounding; Inflammation; Oils; Volatile; Toxicology; Viper Venoms

Figure

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  Fig. 1 Effect of MLO + OVEO solution against formalin-induced pain according pretreatment period. The data represent mean ± SEM (n = 6 for each group). MLO: Macrovipera lebetina obtusa, OVEO: Origanum vulgare L. essential oil, SEM: standard error of mean.

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  Fig. 2 Comparison of the analgesic effect of MLO + OVEO with the effect of standard analgesics. The data represent mean ± SEM (n = 6 for each group). MLO: Macrovipera lebetina obtusa, OVEO: Origanum vulgare L. essential oil, SEM: standard error of mean.

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  Fig. 3 The dynamics of anti-inflammatory effect of liqiud preparation on carrageenan-induced edema. The “0” level means volume of intact paw taken as 100%. Carrageenan – carrageenan induced edema; Diclofenac – Diclofenac solution, IP; MLO + OVEO solution, IP. Data represent mean ± SEM (n = 6 for each group). MLO: Macrovipera lebetina obtusa, OVEO: Origanum vulgare L. essential oil, SEM: standard error of mean, ns: not significant, IP: intraperitoneal. ****P < 0.0001.

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  Fig. 4 Effect of the combined preparation and standard painkillers in hot plate test in mice. The data represent mean ± SEM (n = 6 for each group). MLO: Macrovipera lebetina obtusa, OVEO: Origanum vulgare L. essential oil, SEM: standard error of mean, ns: not significant.

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  Fig. 5 The toxicity LD50 of the combined solution (IP injection). The therapeutic dose was taken as arbitrary units = 1. The 1, 2-, 3-, 4-, and 5-times higher doses were tested in 5 experimental groups (n = 6 for each group). MLO: Macrovipera lebetina obtusa, OVEO: Origanum vulgare L. essential oil, IP: intraperitoneal.

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  Fig. 6 The analgesic effect of the ointment for the entire period of pain development during formalin biphasic action (A) and in the second phase (B). The data represent mean ± SEM (n = 6 for each group). MLO: Macrovipera lebetina obtusa, OVEO: Origanum vulgare L. essential oil, SEM: standard error of mean. **P < 0.01; ****P < 0.0001.

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  Fig. 7 Reduction of carrageenan-induced inflammatory edema by ointments. The “0” level means volume of intact paw taken as 100%. Carrageenan – carrageenan-induced edema; Diclofenac – diclofenac ointment; Ointment – ointment with OVEO + MLO; MLO ointment – ointment with MLO venom alone, OVEO ointment – ointment with OVEO alone. Data represent mean ± SEM (n = 6 for each group). MLO: Macrovipera lebetina obtusa, OVEO: Origanum vulgare L. essential oil, SEM: standard error of mean, ns: not significant. ***P < 0.001; ****P < 0.0001.

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  Fig. 8 Effect of the IPL injection of SR144528 on the antinociceptive activity of ointment in the FT during the entire period of pain development (A) and separately in different phases (B). The data represent mean ± SEM (n = 6 for each group). IPL: intraplantar, FT: formalin test, SEM: standard error of mean, ns: not significant. **P < 0.01; ***P < 0.001; ****P < 0.0001.

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  Fig. 9 Effect of the IPL injection of naloxone on the antinociceptive activity of ointment in the FT during the entire period of pain development (A) and separately in different phases (B). The data represent mean ± SEM (n = 6 for each group). IPL: intraplantar, FT: formalin test, SEM: standard error of mean. **P < 0.01; ****P < 0.0001.

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  Fig. 10 Skin irritation assessment. (A) Before treatment; (B) 7th day after daily treatment of ointment (n = 6 for each group).

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  Fig. 11 The intradermal dynamics of the main component of OVEO, β-caryophyllene, as a penetration enhancer. OVEO: Origanum vulgare L. essential oil, PE: penetration enhancer. Adapted from the article of Tang et al. (Eur J Pharm Sci 2023; 183: 106401) [36].

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