Study on cholinomimetic properties of 2-methoxy-5-trifluoromethylbenzaldehyde as a modulator of contractile activity of smooth muscles of rat caecum
T. Fedirko
, V. Bdzhola
, S. Starosyla
, I. Voiteshenko
, V. Hitchenko, K. Pisnya, K. Zhyshkevych, A. Musіienko
, O. Nyporko
, O. Tsymbalyuk
Abstract: Objective: To evaluate the pharmacological and kinetic effects of 2 methoxy 5 trifluoromethylbenzaldehyde(MTMBA), predicted in silico to activate muscarinic acetylcholine receptors (mAChRs), on the contractile activity of rat caecum smooth muscles. Materials and methods: Experiments were conducted on Wistar rat caecum circular smooth muscle preparations using tenzometric registration in isometric and isotonic modes. Mechanokinetic analysis was performed via the Kosterin-Burdyga method to determine normalized maximal velocities of contraction (Vnc) and relaxation (Vnr). The effects of MTMBA (10-9–10-6 M) were evaluated on acetylcholine-induced contractions, including selective blocking with M2 (AF-DX 116) and M3 (4-DAMP) antagonists. Results: MTMBA (10⁻⁹–10⁻⁶ M) significantly enhanced tonic components of acetylcholine induced contractions, indicating activation of M2 mAChRs. At 10⁻⁹–10⁻⁸ M, MTMBA also increased phase components, likely via M3 mAChR excitation. Across all concentrations, MTMBA altered Vnc and Vnr. The activating effect at 10⁻⁸ M was abolished by AF DX 116 (M2 antagonist) but persisted with 4 DAMP (M3 antagonist), confirming M2 selectivity. Conclusion: MTMBA enhanced acetylcholine induced contractions of rat caecum smooth muscles, demonstrating cholinomimetic properties. At 10⁻⁸ M, it acted as a selective agonist of M2 mAChRs, while at higher concentrations (10⁻⁷–10⁻⁶ M) additional cellular targets may contribute to modulation of intestinal smooth muscle excitability.
Series on Biomechanics, Vol.40, No. 2 (2026), 45-58
DOI: 10.7546/SB.40.02.05.2026
Keywords: acetylcholine; agonist; contraction; Muscarinic acetylcholine receptors; smooth muscles
| References: (click to open/close) | [1] Ishii, M., Kurachi, Y., 2006. Muscarinic acetylcholine receptors. Current Pharmaceutical Design 12, 3573–3581.
[2] Seeger, T., Fedorova, I., Zheng, F., Miyakawa, T., Koustova, E., Gomeza, J., Basile, A. S., Alzheimer, C., Wess, J., 2004. M2 muscarinic acetylcholine receptor knock-out mice show deficits in behavioral flexibility, working memory, and hippocampal plasticity. The Journal of Neuroscience 24, 10117–10127.
[3] Uggenti, C., De Stefano, M. E., Costantino, M., Loreti, S., Pisano, A., Avallone, B., Talora, C., Magnaghi, V., Tata, A. M., 2014. M2 muscarinic receptor activation regulates Schwann cell differentiation and myelin organization. Developmental Neurobiology 74, 676–691.
[4] Cai, Y. Q., Chen, S. R., Han, H. D., Sood, A. K., Lopez-Berestein, G., Pan, H. L., 2009. Role of M2, M3, and M4 muscarinic receptor subtypes in the spinal cholinergic control of nociception revealed using siRNA in rats. Journal of Neurochemistry 111, 1000–1010.
[5] Bernardini, N., Roza, C., Sauer, S. K., Gomeza, J., Wess, J., Reeh, P. W., 2002. Muscarinic M2 receptors on peripheral nerve endings: a molecular target of antinociception. The Journal of Neuroscience 22, RC229.
[6] Chen, S. R., Wess, J., Pan, H. L., 2005. Functional activity of the M2 and M4 receptor subtypes in the spinal cord studied with muscarinic acetylcholine receptor knockout mice. The Journal of Pharmacology and Experimental Therapeutics 313, 765–770.
[7] Lucianò, A. M., Perciballi, E., Fiore, M., Del Bufalo, D., Tata, A. M., 2020. The Combination of the M2 Muscarinic Receptor Agonist and Chemotherapy Affects Drug Resistance in Neuroblastoma Cells. International Journal of Molecular Sciences 21, 8433.
[8] Español, A. J., Salem, A., Di Bari, M., Cristofaro, I., Sanchez, Y., Tata, A. M., Sales, M. E., 2020. The metronomic combination of paclitaxel with cholinergic agonists inhibits triple negative breast tumor progression. Participation of M2 receptor subtype. PLOS ONE 15, e0226450.
[9] Pacini, L., De Falco, E., Di Bari, M., Coccia, A., Siciliano, C., Ponti, D., Pastore, A. L., Petrozza, V., Carbone, A., Tata, A. M., Calogero, A., 2014. M2 muscarinic receptors inhibit cell proliferation and migration in urothelial bladder cancer cells. Cancer Biology & Therapy 15, 1489–1498.
[10] Liao, F., Zheng, Y., Cai, J., Fan, J., Wang, J., Yang, J., Cui, Q., Xu, G., Tang, C., Geng, B., 2015. Catestatin attenuates endoplasmic reticulum induced cell apoptosis by activation type 2 muscarinic acetylcholine receptor in cardiac ischemia/reperfusion. Scientific Reports 5, 16590.
[11] Dolejší, E., Janoušková, A., Jakubík, J., 2024. Muscarinic Receptors in Cardioprotection and Vascular Tone Regulation. Physiological Research 73 (Suppl 1), S389–S400.
[12] Alom, F., Miyakawa, M., Matsuyama, H., Nagano, H., Tanahashi, Y., Unno, T., 2018. Possible antagonistic effects of the TRPC4 channel blocker ML204 on M2 and M3 muscarinic receptors in mouse ileal and detrusor smooth muscles and atrial myocardium. The Journal of Veterinary Medical Science 80, 1407–1415.
[13] Alom, F., Matsuyama, H., Nagano, H., Fujikawa, S., Tanahashi, Y., Unno, T., 2019. Involvement of transient receptor potential melastatin 4 channels in the resting membrane potential setting and cholinergic contractile responses in mouse detrusor and ileal smooth muscles. The Journal of Veterinary Medical Science 81, 217–228.
[14] Griffin, C. S., Bradley, E., Dudem, S., Hollywood, M. A., McHale, N. G., Thornbury, K. D., Sergeant, G. P., 2016. Muscarinic Receptor Induced Contractions of the Detrusor are Mediated by Activation of TRPC4 Channels. The Journal of Urology 196, 1796–1808.
[15] Sá-Caputo, D., Taiar, R., Martins-Anjos, E., Seixas, A., Sartório, A., Sanudo, B., Sonza, A., Amaral, V., Lacerda, A., Gomes-Neto, M., Moura-Filho, O., Oliveira, L., Bachur, J., Bernardo-Filho, M., 2023. Does the mechano-biomodulation vibration lead to biological responses on human beings. Series on Biomechanics 37, 3–17.
[16] Goletiani, C., Nebieridze, N., Mantskava, M., Zhvania, M., Songulashvili, D., 2021. Therapeutic effect of discriminated electric stimuli on the bursts in dissociated cortical culture neural networks. Series on Biomechanics 35, 32–41.
[17] Zholos, A. V., Dryn, D. O., Melnyk, M. I., 2023. General anaesthesia-related complications of gut motility with a focus on cholinergic mechanisms, TRP channels and visceral pain. Frontiers in Physiology 14, 1174655.
[18] Guerriero, C., Manfredelli, M., Matera, C., Iuzzolino, A., Conti, L., Dallanoce, C., De Amici, M., Trisciuoglio, D., Tata, A. M., 2023. M2 Muscarinic Receptor Stimulation Induces Autophagy in Human Glioblastoma Cancer Stem Cells via mTOR Complex-1 Inhibition. Cancers 16, 25.
[19] Intachai, K., C Chattipakorn, S., Chattipakorn, N., Shinlapawittayatorn, K., 2018. Revisiting the Cardioprotective Effects of Acetylcholine Receptor Activation against Myocardial Ischemia/Reperfusion Injury. International Journal of Molecular Sciences 19, 2466.
[20] Vukova, T., Apostolova, S., Georgieva, I., Tzoneva, R., Antonova, N., 2024. Effect of mechanical stress and blood flow on the adhesion of white blood cells to the endothelium in terms of inflammatory processes. Series on Biomechanics 38, 65–73.
[21] Xu, J., Wang, Q., Hübner, H., Hu, Y., Niu, X., Wang, H., Maeda, S., Inoue, A., Tao, Y., Gmeiner, P., Du, Y., Jin, C., Kobilka, B. K., 2023. Structural and dynamic insights into supra-physiological activation and allosteric modulation of a muscarinic acetylcholine receptor. Nature Communications 14, 376.
[22] Kaoullas, M. G., Thal, D. M., Christopoulos, A., Valant, C., 2024. Ligand bias at the muscarinic acetylcholine receptor family: Opportunities and challenges. Neuropharmacology 258, 110092.
[23] Sapundzhi, F., Georgiev, S., Lazarova, M., Popstoilov, M., 2025. Application of artificial intelligence and machine learning to study the structurebiological activity relationship. Series on Biomechanics 39, 74–84.
[24] Kruse, A. C., Kobilka, B. K., Gautam, D., Sexton, P. M., Christopoulos, A., Wess, J., 2014. Muscarinic acetylcholine receptors: novel opportunities for drug development. Nature Reviews Drug Discovery 13, 549–560.
[25] Myslivecek, J., 2022. Multitargeting nature of muscarinic orthosteric agonists and antagonists. Frontiers in Physiology 13, 974160.
[26] Santiago, L. J., Abrol, R., 2019. Understanding G Protein Selectivity of Muscarinic Acetylcholine Receptors Using Computational Methods. International Journal of Molecular Sciences 20, 5290.
[27] Nyporko, A., Tsymbalyuk, O., Voiteshenko, I., Starosyla, S., Protopopov, M., Bdzhola, V., 2023. Computer-aided design of muscarinic acetylcholine receptor M3 inhibitors: Promising compounds among trifluoromethyl containing hexahydropyrimidinones/thiones. Molecular Informatics 42, 2300006.
[28] Nyporko, A., Tsymbalyuk, O., Voiteshenko, I., Starosyla, S., Protopopov, M., Bdzhola, V., 2023. New promising agents against COPD and asthma among the amides of 1-oxo-3-phenyl-isochroman-6-carboxylic acid. Biophysical Bulletin 50, 36–47.
[29] Tsymbalyuk, O., Voiteshenko, I., Starosyla, S., Bdzhola, V., Prykhodko, A., Fedirko, T., Savchenko, O., Bakhina, A., Nyporko, O., 2023. 8-[(4-benzylpiperazin-1-yl)methyl]-3-(2-chlorophenyl)-7-hydroxy-chromen-4-one is an activator of contractile activity of intestinal smooth muscles with reversible M2 cholinomimetic properties. Studia Biologica 17, 15–36.
[30] Burdyga, Th. V., Kosterin, S. A., 1991. Kinetic analysis of smooth muscle relaxation. General Physiology and Biophysics 10, 589–598.
[31] Tsymbalyuk, O., Veklich, T., Rodik, R., Maliuk, O., Karakhim, S., Kalchenko, V., Kosterin, S., 2024. Modulation of myometrium contractive activity and intracellular Ca2+ homeostasis by calix4arene С-1130 selective inhibitor of the sodium pump. Series on Biomechanics 38, 71–89.
[32] Tanahashi, Y., Komori, S., Matsuyama, H., Kitazawa, T., Unno, T., 2021. Functions of Muscarinic Receptor Subtypes in Gastrointestinal Smooth Muscle: A Review of Studies with Receptor-Knockout Mice. International Journal of Molecular Sciences 22, 926.
[33] Kang, H., Kim, J., Park, C. H., Jeong, B., So, I., 2024. Direct modulation of TRPC ion channels by Gα proteins. Frontiers in Physiology 15, 1362987.
[34] Sakamoto, T., Unno, T., Kitazawa, T., Taneike, T., Yamada, M., Wess, J., Nishimura, M., Komori, S., 2007. Three distinct muscarinic signalling pathways for cationic channel activation in mouse gut smooth muscle cells. The Journal of Physiology 582, 41–61.
[35] Cohen-Armon, M., 2023. Are Voltage Sensors Really Embedded in Muscarinic Receptors?. International Journal of Molecular Sciences 24, 7538.
[36] Tauber, M., Ben-Chaim, Y., 2024. Voltage Sensors Embedded in G Protein-Coupled Receptors. International Journal of Molecular Sciences 25, 5295.
|
|
| Date published: 2026-07-17
(Price of one pdf file: 25.00 EUR)
