Insomnia model selection for comparative evaluation of melatonin-based preparations

N.V. Demakova1, ORCID 0000-0002-2145-9289, 
M.A. Korotyhina1,
ORCID 0000-0002-2368-3268
Yu.I. Sysoev1–3, ORCID 0000-0003-4199-5318
A.V. Karshin4, ORCID 0000-0001-8145-9603
N.A. Anisimova1, ORCID 0000-0002-2620-2976
D.Yu. Ivkin1, ORCID 0000-0001-9273-6864
1Saint Petersburg State Chemical Pharmaceutical University of the Ministry of Health Care of Russia,
197376, Russian Federation, Saint Petersburg, Professora Popova str., 14;
2Institute of Translational Biomedicine, Saint Petersburg State University,
199034, Russian Federation, Saint Petersburg, Universitetskaya emb., 7–9;
3Pavlov Institute of Physiology, Russian Academy of Sciences (RAS),
199034, Russian Federation, Saint Petersburg , Makarova emb., 6;
4OOO «Farmamed»,194292, Russian Federation, Saint Petersburg, Domostroitelnaya st., 16, lit. E


Currently insomnia is prevalent among 30–35% of population. As it stands now the pharmaceutical market for insomnia drugs is practically not replenished, and therefore, it is urgent to create new, more effective and / or safer drugs.

The aim of the presented work is to select and test the optimal insomnia model for a comparative assessment of the effectiveness of melatonin preparations.

A number of existing Russian and foreign models of insomnia and methods of its study were assessed, including the use of such a database of medical and biological publications as PubMed. The considered models were divided into three large blocks: models associated with stress factors, pharmacological models and genetic ones.  Animals such as rabbits, rats, mice, Danio rerio and Drosophila sp. were used as test systems for studies on models of insomnia from the analyzed literature sources. As the most suitable for the study of the comparative pharmacodynamics of a new drug based on melatonin and herbal components in terms of the severity of the hypnotic effect, rodent cage change insomnia model was chosen. Its essence boils down to changing the usual cage in sexually mature males to a cage with stale bedding containing an olfactory trace of another gender-identical animal, which acts as a stress factor in the form of expectation of aggression and allows creating the necessary conditions for conducting an experiment.

The results of a study of a melatonin preparation with a phytocomplex containing extracts of raw materials of Valeriana officinalis, Leonurus cardiaca, Crataegus sanguinea, Mentha piperita (+ oleum Menthae piperitae) indicates a significant improvement in the ratio of the theta and delta rhythm indices of animals and the alpha rhythm index, which are close to the values of the control group animals. On the basis of the EEG study, it can be concluded that the tested combination drug is more effective in comparison with the reference melatonin. Rodent cage change insomnia model is an effective and easily reproducible model which can be used not only for comparative assessment of melatonin preparations efficacy, but also for other pharmacological agents for the treatment of insomnia.

Full text avaliable in Russain only 

Author’s Contribution

M.A. Korotyhina – carried out the experiment, discussed the results, worked with literature sources, wrote the manuscript, corrected the text in accordance with the requirements of the journal
N.V. Demakova – carried out the experiment, discussed the results, worked with literature sources, wrote the manuscript, corrected the text in accordance with the requirements of the journal
Yu.I. Sysoev – carried out the experiment, performed ECOG electrode implantation, analysed the data, discussed the results, wrote the manuscript
A.V. Karshin – provided the laboratory with tested substance of melatonin + phytocomplex, confirmed the experiment design, approved the final version of the article for publication
N.A. Anisimova – literature sources analysis
D.Yu. Ivkin – designed the experiment, summarized the material, wrote the manuscript, approved the final version of the article for publication


The study was performed without external funding


Conflict of interest

The authors declare no conflicts of interest.


  1. Bathory, E., & Tomopoulos, S. (2017). Sleep Regulation, Physiology and Development, Sleep Duration and Patterns, and Sleep Hygiene in Infants, Toddlers, and Preschool-Age Children. Current problems in pediatric and adolescent health care, 47(2), 29–42.
  2. Revel, F. G., Gottowik, J., Gatti, S., Wettstein, J. G., & Moreau, J. L. (2009). Rodent models of insomnia: a review of experimental procedures that induce sleep disturbances. Neuroscience and biobehavioral reviews, 33(6), 874–899.
  3. Morin, C. M., Drake, C. L., Harvey, A. G., Krystal, A. D., Manber, R., Riemann, D., & Spiegelhalder, K. (2015). Insomnia disorder. Nature reviews. Disease primers, 1, 15026.
  4. Стрыгин К.Н., Полуэктов М.Г. ИНСОМНИЯ. Медицинский Совет. 2017;(1S):52-58. [Strygin K.N., Poluektov M.G. INSOMNIYA. Medicinskij Sovet. 2017;(1S):52-58. (In Russ.)]
  5. Кемстач В.В., Коростовцева Л.С., Алёхин А.Н., и др. Исследования психофизиологических аспектов и этиопатогенеза инсомнии: российские и зарубежные подходы // Вестник Российского университета дружбы народов. Серия: Психология и педагогика. - 2020. - Т. 17. - №2. - C. 288-309. doi: 10.22363/2313-1683-2020-17-2-288-309 [Kemstach V.V., Korostovceva L.S., Alyohin A.N., i dr. Issledovaniya psihofiziologicheskih aspektov i etiopatogeneza insomnii: rossijskie i zarubezhnye podhody // Vestnik Rossijskogo universiteta druzhby narodov. Seriya: Psihologiya i pedagogika. - 2020. - V. 17. - №2. - p. 288-309. (In Russ.)]
  6. Bonnet, C., Léger, L., Baubet, V., Debilly, G., & Cespuglio, R. (1997). Influence of a 1 h immobilization stress on sleep states and corticotropin-like intermediate lobe peptide (CLIP or ACTH18-39, Ph-ACTH18-39) brain contents in the rat. Brain research, 751(1), 54–63.
  7. Rampin, C., Cespuglio, R., Chastrette, N., & Jouvet, M. (1991). Immobilisation stress induces a paradoxical sleep rebound in rat. Neuroscience letters, 126(2), 113–118.
  8. Chang, F. C., & Opp, M. R. (2002). Role of corticotropin-releasing hormone in stressor-induced alterations of sleep in rat. American journal of physiology. Regulatory, integrative and comparative physiology, 283(2), R400–R407.
  9. Grahnstedt, S., & Ursin, R. (1985). Platform sleep deprivation affects deep slow wave sleep in addition to REM sleep. Behavioural brain research, 18(3), 233–239.
  10. Ramm, P., & Smith, C. T. (1990). Rates of cerebral protein synthesis are linked to slow wave sleep in the rat. Physiology & behavior, 48(5), 749–753.
  11. Machado, R. B., Suchecki, D., & Tufik, S. (2006). Comparison of the sleep pattern throughout a protocol of chronic sleep restriction induced by two methods of paradoxical sleep deprivation. Brain research bulletin, 70(3), 213–220.
  12. Shinomiya, K., Shigemoto, Y., Okuma, C., Mio, M., & Kamei, C. (2003). Effects of short-acting hypnotics on sleep latency in rats placed on grid suspended over water. European journal of pharmacology, 460(2-3), 139–144.
  13. Shinomiya, K., Shigemoto, Y., Omichi, J. et al. Effects of three hypnotics on the sleep-wakefulness cycle in sleep-disturbed rats. Psychopharmacology 173, 203–209 (2004).
  14. Shinomiya, K., Fujimura, K., Kim, Y., & Kamei, C. (2005). Effects of valerian extract on the sleep-wake cycle in sleep-disturbed rats. Acta medica Okayama, 59(3), 89–92.
  15. Pawlyk, A. C., Morrison, A. R., Ross, R. J., & Brennan, F. X. (2008). Stress-induced changes in sleep in rodents: models and mechanisms. Neuroscience and biobehavioral reviews, 32(1), 99–117.
  16. Sanford, L. D., Fang, J., & Tang, X. (2003). Sleep after differing amounts of conditioned fear training in BALB/cJ mice. Behavioural brain research, 147(1-2), 193–202.
  17. Pawlyk, A. C., Jha, S. K., Brennan, F. X., Morrison, A. R., & Ross, R. J. (2005). A rodent model of sleep disturbances in posttraumatic stress disorder: the role of context after fear conditioning. Biological psychiatry, 57(3), 268–277.
  18. Huang, M. P., Radadia, K., Macone, B. W., Auerbach, S. H., & Datta, S. (2010). Effects of eszopiclone and zolpidem on sleep-wake behavior, anxiety-like behavior and contextual memory in rats. Behavioural brain research, 210(1), 54–66.
  19. Amici, R., Domeniconi, R., Jones, C. A., Morales-Cobas, G., Perez, E., Tavernese, L., Torterolo, P., Zamboni, G., & Parmeggiani, P. L. (2000). Changes in REM sleep occurrence due to rhythmical auditory stimulation in the rat. Brain research, 868(2), 241–250.
  20. Khazan, N., & Sawyer, C. H. (1963). “Rebound” Recovery from Deprivation of Paradoxical Sleep in the Rabbit. Proceedings of the Society for Experimental Biology and Medicine, 114(2), 536–539.
  21. Zhang, Z. Q., Degejin, Geng, D., Zhang, Q., Tian, Y., Xi, Y., Wang, W. Q., Tang, H. Q., Xu, B., Lin, H. Y., & Sun, Y. K. (2016). Pharmacodynamic study on insomnia-curing effects of Shuangxia Decoction in Drosophila melanogaster. Chinese journal of natural medicines, 14(9), 653–660.
  22. McKenna, J. T., Gamble, M. C., Anderson-Chernishof, M. B., Shah, S. R., McCoy, J. G., & Strecker, R. E. (2019). A rodent cage change insomnia model disrupts memory consolidation. Journal of sleep research, 28(2), e12792.
  23. Iturra-Mena, A. M., Arriagada-Solimano, M., Luttecke-Anders, A., & Dagnino-Subiabre, A. (2018). Effects of prenatal stress on anxiety- and depressive-like behaviours are sex-specific in prepubertal rats. Journal of neuroendocrinology, 30(7), e12609.
  24. Datta, S., Patterson, E. H., Vincitore, M., Tonkiss, J., Morgane, P. J., & Galler, J. R. (2000). Prenatal protein malnourished rats show changes in sleep/wake behavior as adults. Journal of sleep research, 9(1), 71–79.
  25. Sylvester, L., Kapron, C. M., & Smith, C. (2000). In utero ethanol exposure decreases rapid eye movement sleep in female Sprague-Dawley rat offspring. Neuroscience letters, 289(1), 13–16.
  26. Alföldi, P., Rubicsek, G., Cserni, G. et al. Brain and core temperatures and peripheral vasomotion during sleep and wakefulness at various ambient temperatures in the rat. Pflugers Arch. 417, 336–341 (1990).
  27. Amici, R., Zamboni, G., Perez, E., Jones, C. A., Toni, I., I, Culin, F., & Parmeggiani, P. L. (1994). Pattern of desynchronized sleep during deprivation and recovery induced in the rat by changes in ambient temperature. Journal of sleep research, 3(4), 250–256.
  28. Landis, C. A., Levine, J. D., & Robinson, C. R. (1989). Decreased slow-wave and paradoxical sleep in a rat chronic pain model. Sleep, 12(2), 167–177.
  29. Ko, C. H., Koon, C. M., Yu, S. L., Lee, K. Y., Lau, C. B., Chan, E. H., Wing, Y. K., Fung, K. P., & Leung, P. C. (2016). Hypnotic effects of a novel anti-insomnia formula on Drosophila insomnia model. Chinese journal of integrative medicine, 22(5), 335–343.
  30. Veatch L. M. (2006). Disruptions in sleep time and sleep architecture in a mouse model of repeated ethanol withdrawal. Alcoholism, clinical and experimental research, 30(7), 1214–1222.
  31. Kubota, T., De, A., Brown, R. A., Simasko, S. M., & Krueger, J. M. (2002). Diurnal effects of acute and chronic administration of ethanol on sleep in rats. Alcoholism, clinical and experimental research, 26(8), 1153–1161.
  32. Wang, Q., Ren, X., Zhang, X., Wang, G., Xu, H., Deng, N., Liu, T., & Peng, Z. (2020). Therapeutic Effect of Berberine on Insomnia Rats by ErbB Signaling Pathway. Medical science monitor : international medical journal of experimental and clinical research, 26, e921831.
  33. Xu, Y., Li, X., Man, D., Su, X., & A, G. (2020). iTRAQ-based proteomics analysis on insomnia rats treated with Mongolian medical warm acupuncture. Bioscience reports, 40(5), BSR20191517.
  34. Ren, X. J., Wang, G. Y., Zhang, X. P., Wang, Q. Q., & Peng, Z. P. (2020). Sedative and Hypnotic Effects and Transcriptome Analysis of Polygala tenuifolia in Aged Insomnia Rats. Chinese journal of integrative medicine, 26(6), 434–441.
  35. Dringenberg, H. C., Hargreaves, E. L., Baker, G. B., Cooley, R. K., & Vanderwolf, C. H. (1995). p-chlorophenylalanine-induced serotonin depletion: reduction in exploratory locomotion but no obvious sensory-motor deficits. Behavioural brain research, 68(2), 229–237.
  36. Prober, D. A., Rihel, J., Onah, A. A., Sung, R. J., & Schier, A. F. (2006). Hypocretin/orexin overexpression induces an insomnia-like phenotype in zebrafish. The Journal of neuroscience : the official journal of the Society for Neuroscience, 26(51), 13400–13410.
  37. Ko, C. H., Koon, C. M., Yu, S. L., Lee, K. Y., Lau, C. B., Chan, E. H., Wing, Y. K., Fung, K. P., & Leung, P. C. (2016). Hypnotic effects of a novel anti-insomnia formula on Drosophila insomnia model. Chinese journal of integrative medicine, 22(5), 335–343.
  38. Belfer, S. J., Bashaw, A. G., Perlis, M. L., & Kayser, M. S. (2021). A Drosophila model of sleep restriction therapy for insomnia. Molecular psychiatry, 26(2), 492–507.
  39. Chesselet, M. F., Richter, F., Zhu, C., Magen, I., Watson, M. B., & Subramaniam, S. R. (2012). A progressive mouse model of Parkinson's disease: the Thy1-aSyn ("Line 61") mice. Neurotherapeutics : the journal of the American Society for Experimental NeuroTherapeutics, 9(2), 297–314.
  40. McDowell, K. A., Shin, D., Roos, K. P., & Chesselet, M. F. (2014). Sleep dysfunction and EEG alterations in mice overexpressing alpha-synuclein. Journal of Parkinson's disease, 4(3), 531–539.
  41. Medeiros, D. C., Lopes Aguiar, C., Moraes, M., & Fisone, G. (2019). Sleep Disorders in Rodent Models of Parkinson's Disease. Frontiers in pharmacology, 10, 1414.
  42. Sack, R. L., Auckley, D., Auger, R. R., Carskadon, M. A., Wright, K. P., Jr, Vitiello, M. V., Zhdanova, I. V., & American Academy of Sleep Medicine (2007). Circadian rhythm sleep disorders: part I, basic principles, shift work and jet lag disorders. An American Academy of Sleep Medicine review. Sleep, 30(11), 1460–1483.
  43. Schulz, P., & Steimer, T. (2009). Neurobiology of circadian systems. CNS drugs, 23 Suppl 2, 3–13.
  44. Jan, J. E., Reiter, R. J., Wasdell, M. B., & Bax, M. (2009). The role of the thalamus in sleep, pineal melatonin production, and circadian rhythm sleep disorders. Journal of pineal research, 46(1), 1–7.
  45. Шекунова Е.В., Ковалева М.А., Макарова М.Н., Макаров В.Г. ВЫБОР ДОЗЫ ПРЕПАРАТА ДЛЯ ДОКЛИНИЧЕСКОГО ИССЛЕДОВАНИЯ: МЕЖВИДОВОЙ ПЕРЕНОС ДОЗ // Ведомости Научного центра экспертизы средств медицинского применения. 2020. Т. 10. № 1. С. 19-28. [Shekunova E.V., Kovaleva M.A., Makarova M.N., Makarov V.G. VYBOR DOZY PREPARATA DLYA DOKLINICHESKOGO ISSLEDOVANIYA: MEZHVIDOVOJ PERENOS DOZ // Vedomosti Nauchnogo centra ekspertizy sredstv medicinskogo primeneniya. 2020. T. 10. № 1. S. 19-28. (In Russ.)]
  46. Paxinos G., Watson C.The rat brain in stereotaxic coordinates. 7th edition. Acad. Press. 2013.
  47. Жирмунская Е.А. Клиническая энцефалография. М.: МЭЙБИ, 1991. 118 с. [Zhirmunskaya E.A. Klinicheskaya encefalografiya. M.: MEJBI, 1991. 118 p. (In Russ.)]
  48. Федотова, И. Р. Исследование структуры гиппокампальной тета-активности и ее регуляции при ориентировочном поведении крысы : специальность 03.00.13 : диссертация на соискание ученой степени кандидата биологических наук / Федотова Ирина Ростиславовна. – Москва, 2000. – 182 с.[Fedotova, I. R. Issledovanie struktury gippokampal'noj teta-aktivnosti i ee regulyacii pri orientirovochnom povedenii krysy : special'nost' 03.00.13 : dissertaciya na soiskanie uchenoj stepeni kandidata biologicheskih nauk / Fedotova Irina Rostislavovna. – Moskva, 2000. – 182 p. (In Russ.)]
  49. Физиология сна: Учебное пособие для ветеринарных врачей, зооинженеров, студентов факультета ветеринарной медицины, зооинженерного факультета и слушателей ФПК / В.В. Ковзов, В.К. Гусаков, А.В. Островский. – Витебск: УО ВГАВМ, 2005. – 59 с. [Fiziologiya sna: Uchebnoe posobie dlya veterinarnyh vrachej, zooinzhenerov, studentov fakul'teta veterinarnoj mediciny, zooinzhenernogo fakul'teta i slushatelej FPK / V.V. Kovzov, V.K. Gusakov, A.V. Ostrovskij. – Vitebsk: UO VGAVM, 2005. – 59 p. (In Russ.)]
  50. Barbosa-Méndez, S., & Salazar-Juarez, A. (2020). Melatonin does not produce sedation in rats: A chronobiological study. Chronobiology international, 37(3), 353–374.

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