pharmjournalРазработка и регистрация лекарственных средствDrug development & registration2305-20662658-5049LLC «CPHA»10.33380/2305-2066-2024-13-2-1772pharmjournal-1834Research ArticleФАРМАЦЕВТИЧЕСКАЯ ТЕХНОЛОГИЯPHARMACEUTICAL TECHNOLOGYАлгоритм разработки комбинированной фармацевтической субстанции растительного происхождения с антимикробным действием: от научного поиска до лекарственной формыAlgorithm for the development of a multicomponent pharmaceutical substance of plant origin with antimicrobial action: from science search to dosage formhttps://orcid.org/0000-0001-7179-8062ФилатовВ. А.FilatovV. A.

119192, г. Москва, Ломоносовский пр-т., д. 27, корп. 1

27/1, Lomonosovsky prospect, Moscow, 119192

filatovviktor097@gmail.comhttps://orcid.org/0000-0003-0385-3474КулякО. Ю.KulyakO. Yu.

119192, г. Москва, Ломоносовский пр-т., д. 27, корп. 1

27/1, Lomonosovsky prospect, Moscow, 119192

https://orcid.org/0000-0003-0068-2788КалениковаЕ. И.KalenikovaE. I.

119192, г. Москва, Ломоносовский пр-т., д. 27, корп. 1

27/1, Lomonosovsky prospect, Moscow, 119192

Федеральное государственное бюджетное образовательное учреждение высшего образования «Московский государственный университет имени М. В. Ломоносова» (МГУ имени М. В. Ломоносова)РоссияLomonosov Moscow State UniversityRussian Federation20242405202413294105Copyright © Филатов В.А., Куляк О.Ю., Каленикова Е.И., 20242024Филатов В.А., Куляк О.Ю., Каленикова Е.И.Filatov V.A., Kulyak O.Y., Kalenikova E.I.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.pharmjournal.ru/jour/article/view/1834Введение

Введение. Несмотря на доказанную клиническую эффективность противогрибковых и противовоспалительных лекарственных препаратов для профилактики и лечения себорейного дерматита, актуален поиск новых мишеней и разработка новых субстанций с благоприятным действием на микрофлору кожи головы, низким риском развития резистентности бактерий и побочных эффектов.

Цель

Цель. Поэтапная разработка антимикробной комбинированной фармацевтической субстанции растительного происхождения: от научного поиска перспективных субстанций, анализа их состава методом ГХ-МС, оценки in silico степени сродства отдельных компонентов к патогенетическим мишеням, выбора оптимального состава комбинированной субстанции по результатам тестирования in vitro антимикробного действия до создания препарата на ее основе в форме лекарственного шампуня для терапии себорейного дерматита.

Материалы и методы

Материалы и методы. Объекты исследования: эфирное масло листьев чайного дерева, 1,8-цинеол, α-(-)-бисаболол и комбинированная субстанция на их основе в массовом соотношении 1 : 1 : 1. Методы: молекулярный докинг (AutoDock, версия 4.2), прогноз фармакологической активности (Phyto4Health), ТСХ, ГХ-МС, оценка антимикробной активности in vitro.

Результаты и обсуждение

Результаты и обсуждение. По результатам литературного поиска были выбраны 3 перспективных вещества для разработки комбинированной субстанции растительного происхождения: эфирное масло листьев чайного дерева, 1,8-цинеол и α-(-)-бисаболол. На основании молекулярного докинга предсказано таргетированное действие компонентов эфирного масла листьев чайного дерева, 1,8-цинеола и α-(-)-бисаболола на домены в составе ABC-транспортеров мембраны микроорганизмов, участвующих в патогенезе себорейного дерматита, и обоснована возможность их применения для терапии. Разработана комбинированная субстанция на основе эфирного масла листьев чайного дерева, 1,8-цинеола и α-(-)-бисаболола в массовом соотношении 1 : 1 : 1. Проведена оценка подлинности субстанции методами ТСХ и ГХ-МС, и в ее составе количественно выявлены 15 терпенов с преобладанием терпинен-4-ола (16,98 %), 1,8-цинеола (25,63 %) и α-(-)-бисаболола (27,67 %). Установлена синергетическая и таргетированная антимикробная активность субстанции в отношении Staphylococcus epidermidis, Staphylococcus aureus, Candida albicans и Malassezia furfur в сравнении с бензалкония хлоридом, кетоконазолом и климбазолом. На основе изученной субстанции разработан состав нового лекарственного шампуня, обладающего выраженным противогрибковым эффектом (более 99 %) в отношении M. furfur без видимого подавления нормальной микрофлоры. Для новой субстанции растительного происхождения и лекарственного шампуня оценены показатели качества в соответствии с требованиями Государственной фармакопеи.

Заключение

Заключение. Разработана субстанция растительного происхождения, обладающая синергетической и таргетированной антимикробной активностью и представляющая интерес для дальнейшего изучения в качестве лекарственного средства и активной фармацевтической субстанции новых продуктов для лечения себорейного дерматита.

Introduction

Introduction. Despite the proven clinical efficacy of antifungal and anti-inflammatory drugs for the prevention and treatment of seborrheic dermatitis, the search for new targets and the development of new substances with a beneficial effect on the scalp microflora, with a low risk of antimicrobial resistance and adverse effects, are relevant.

Aim

Aim. Development of the antimicrobial multicomponent pharmaceutical substance of plant origin in stages: from a literature search for promising substances, analysis of their composition by GC-MS, in silico evaluation of the affinity of individual components to pathogenetic targets, selection of the optimal composition of a multicomponent substance based on the results of in vitro research of antimicrobial action and the making of a medical dosage form based on it – a medicinal shampoo for the treatment of seborrheic dermatitis.

Materials and methods

Materials and methods. Objects of research: tea tree essential oil, 1,8-cineole, α-(-)-bisabolol and the multicomponent substance based on them. Methods: molecular docking (AutoDock version 4.2), prediction of pharmacological activity (Phyto4Health), TLC, GC-MS, study of antimicrobial activity in vitro.

Results and discussion

Results and discussion. Based on the results of a literature search, 3 promising substances were selected for the development of a multicomponent plant-based substance: tea tree essential oil, 1,8-cineole and α-(-)-bisabolol. Molecular docking predicted the targeted activity of the phytochemicals of tea tree essential oil, 1,8-cineole and α-(-)-bisabolol on the domains of ABC-transporters of microorganisms involved in the pathogenesis of seborrheic dermatitis and justified the possibility of use for therapy. The multicomponent substance has been developed based on tea tree essential oil, 1,8-cineole and α-(-)-bisabolol in a mass ratio of 1 : 1 : 1. The qualitative composition of the substance was assessed by TLC and GC-MS methods, and 15 terpenes were quantitatively identified in its composition with a predominance of terpinen-4-ol (16.98 %), 1,8-cineole (25.63 %) and α-(-)-bisabolol (27.67 %). The synergistic antimicrobial activity of the substance has been established against S. epidermidis, S. aureus, C. albicans and M. furfur in comparison with benzalkonium chloride, ketoconazole and climbazole. The composition of a new medical shampoo based on the investigated substance has been developed, which has a pronounced antifungal effect (more than 99.0 %) against M. furfur without visible suppression of normal microflora. For the novel substance of plant origin and medical shampoo, quality parameters were assessed in accordance with the Russian Pharmacopoeia of XIV edition.

Conclusion

Conclusion. A substance of plant origin with synergistic and targeted antimicrobial activity has been developed. It has an interest for further study as a drug and API for new products for the treatment of seborrheic dermatitis.

разработкасеборейный дерматитчайное дерево18-цинеолбисабололантимикробное действиелекарственный шампуньдокингфармацевтический анализТСХГХ-МСseborrheic dermatitistea tree oileucalyptolbisabololantimicrobial activitymedical shampoodockingTLCGC-MSРезультаты работы получены с использованием оборудования ФГБОУ ВО «Московский государственный университет имени М. В. Ломоносова», ФГАОУ ВО «Казанский (Приволжский) федеральный университет» и ЦКП НО «Арктика» ФГАОУ ВО «Северный (Арктический) федеральный университет имени М. В. Ломоносова» при административной и финансовой поддержке ООО «Сплат Глобал» и Московского филиала АО «Скайлаб».The results of the research were obtained using the equipment of the Federal State Budget Educational Institution of Higher Education M. V. Lomonosov Moscow State University (Lomonosov MSU), Federal State Autonomous Educational Institution of Higher Education "Kazan (Volga Region) Federal University" and the Central Public Educational Institution "Arctic" of the Federal State Autonomous Educational Institution of Higher Education "Northern (Arctic) Federal University named M. V. Lomonosov" with the administrative and financial support of LLC "Splat Global" and the Moscow branch of JSC "Skylab".ReferencesKatiyar C., Gupta A., Kanjilal S., Katiyar S. Drug discovery from plant sources: An integrated approach. AYU (An International Quarterly Journal of Research in Ayurveda). 2012;33(1):10–19. DOI: 10.4103/0974-8520.100295.Katiyar C., Gupta A., Kanjilal S., Katiyar S. Drug discovery from plant sources: An integrated approach. AYU (An International Quarterly Journal of Research in Ayurveda). 2012;33(1):10–19. DOI: 10.4103/0974-8520.100295.Earm K., Earm Y. E. Integrative approach in the era of failing drug discovery and development. Integrative Medicine Research. 2014;3(4):211–216. DOI: 10.1016/j.imr.2014.09.002.Earm K., Earm Y. E. Integrative approach in the era of failing drug discovery and development. Integrative Medicine Research. 2014;3(4):211–216. DOI: 10.1016/j.imr.2014.09.002.Pinzi L., Rastelli G. Molecular Docking: Shifting Paradigms in Drug Discovery. International Journal of Molecular Sciences. 2019;20(18):4331. DOI: 10.3390/ijms20184331.Pinzi L., Rastelli G. Molecular Docking: Shifting Paradigms in Drug Discovery. International Journal of Molecular Sciences. 2019;20(18):4331. DOI: 10.3390/ijms20184331.Choeng W. K., Yeung C. K., Torsekar R. G., Suh D. H., Ungpakorn R., Widaty S., Azizan N. Z., Gabriel M. T., Tran H. K., Chong W. S., Shih I.-H., Dall’Oglio F., Micali G. Treatment of seborrhoeic dermatitis in Asia: a consensus guide. Skin Appendage Disorders. 2016;1(4):187–196. DOI: 10.1159/000444682.Choeng W. K., Yeung C. K., Torsekar R. G., Suh D. H., Ungpakorn R., Widaty S., Azizan N. Z., Gabriel M. T., Tran H. K., Chong W. S., Shih I.-H., Dall’Oglio F., Micali G. Treatment of seborrhoeic dermatitis in Asia: a consensus guide. Skin Appendage Disorders. 2016;1(4):187–196. DOI: 10.1159/000444682.Gupta A. K., Madzia S. E., Batra R. Etiology and management of Seborrheic dermatitis. Dermatology. 2004;208(2):89–93. DOI: 10.1159/000076478.Gupta A. K., Madzia S. E., Batra R. Etiology and management of Seborrheic dermatitis. Dermatology. 2004;208(2):89–93. DOI: 10.1159/000076478.Borda L. J., Wikramanayake T. C. Seborrheic Dermatitis and Dandruff: A Comprehensive Review. Journal of Clinical and Investigative Dermatology. 2015;3(2):10. DOI: 10.13188/2373-1044.1000019.Borda L. J., Wikramanayake T. C. Seborrheic Dermatitis and Dandruff: A Comprehensive Review. Journal of Clinical and Investigative Dermatology. 2015;3(2):10. DOI: 10.13188/2373-1044.1000019.Adalsteinsson J. A., Kaushik S., Muzumdar S., Guttman-Yassky E., Ungar J. An update on the microbiology, immunology and genetics of seborrheic dermatitis. Experimental Dermatology. 2020;29(5):481–489. DOI: 10.1111/exd.14091.Adalsteinsson J. A., Kaushik S., Muzumdar S., Guttman-Yassky E., Ungar J. An update on the microbiology, immunology and genetics of seborrheic dermatitis. Experimental Dermatology. 2020;29(5):481–489. DOI: 10.1111/exd.14091.Polonskaya A. S., Shatokhina E. A., Kruglova L. S. Seborrheic dermatitis: current ideas of the etiology, pathogenesis, and treatment approaches. 2020;19(4):451–458. (In Russ.) DOI: 10.17116/klinderma202019041451.Polonskaya A. S., Shatokhina E. A., Kruglova L. S. Seborrheic dermatitis: current ideas of the etiology, pathogenesis, and treatment approaches. 2020;19(4):451–458. (In Russ.) DOI: 10.17116/klinderma202019041451.Tao R., Li R., Wang R. Skin microbiome alterations in seborrheic dermatitis and dandruff: A systematic review. Experimental Dermatology. 2021;30(10):1546–1553. DOI: 10.1111/exd.14450.Tao R., Li R., Wang R. Skin microbiome alterations in seborrheic dermatitis and dandruff: A systematic review. Experimental Dermatology. 2021;30(10):1546–1553. DOI: 10.1111/exd.14450.Leong C., Chan J. W. K., Lee S. M., Lam Y. I., Goh J. P. Z., Ianiri G., Dawson T. L. Azole resistance mechanisms in pathogenic Malassezia furfur. Antimicrobial Agents and Chemotherapy. 2021;65(5):1975–2000. DOI: 10.1128/AAC.01975-20.Leong C., Chan J. W. K., Lee S. M., Lam Y. I., Goh J. P. Z., Ianiri G., Dawson T. L. Azole resistance mechanisms in pathogenic Malassezia furfur. Antimicrobial Agents and Chemotherapy. 2021;65(5):1975–2000. DOI: 10.1128/AAC.01975-20.Bukvić Mokos Z., Kralj M., Basta-Juzbašić A., Lakoš Jukić I. Seborrheic dermatitis: an update. Acta Dermatovenerologica Croatica. 2012;20(2):98–104.Bukvić Mokos Z., Kralj M., Basta-Juzbašić A., Lakoš Jukić I. Seborrheic dermatitis: an update. Acta Dermatovenerologica Croatica. 2012;20(2):98–104.Reuter J., Merfort I., Schempp C. M. Botanicals in dermatology: an evidence-based review. American Journal of Clinical Dermatology. 2010;11(4):247–267. DOI: 10.2165/11533220-000000000-00000.Reuter J., Merfort I., Schempp C. M. Botanicals in dermatology: an evidence-based review. American Journal of Clinical Dermatology. 2010;11(4):247–267. DOI: 10.2165/11533220-000000000-00000.Olisova O. Y., Snarskaya E. S., Gladko V. V., Burova E. P. Russian traditional medicine in dermatology. Clinics in Dermatology. 2018;36(3):325–337. DOI: 10.1016/j.clindermatol.2018.03.007.Olisova O. Y., Snarskaya E. S., Gladko V. V., Burova E. P. Russian traditional medicine in dermatology. Clinics in Dermatology. 2018;36(3):325–337. DOI: 10.1016/j.clindermatol.2018.03.007.Karimi A., Majlesi M., Rafieian-Kopaei M. Herbal versus synthetic drugs; beliefs and facts. Journal of Nephropharmacology. 2015;4(1):27–30.Karimi A., Majlesi M., Rafieian-Kopaei M. Herbal versus synthetic drugs; beliefs and facts. Journal of Nephropharmacology. 2015;4(1):27–30.Lynch N., Berry D. Differences in perceived risks and benefits of herbal, over-the-counter conventional, and prescribed conventional, medicines, and the implications of this for the safe and effective use of herbal products. Complementary Therapies in Medicine. 2007;15(2):84–91. DOI: 10.1016/j.ctim.2006.06.007.Lynch N., Berry D. Differences in perceived risks and benefits of herbal, over-the-counter conventional, and prescribed conventional, medicines, and the implications of this for the safe and effective use of herbal products. Complementary Therapies in Medicine. 2007;15(2):84–91. DOI: 10.1016/j.ctim.2006.06.007.Enioutina E. Yu., Teng L., Fateeva T. V., Brown J. C. S., Job K. M., Bortnikova V. V., Krepkova L. V., Gubarev M. I., Sherwin C. M. T. Phytotherapy as an alternative to conventional antimicrobials: combating microbial resistance. Expert Review of Clinical Pharmacology. 2017;10(11):1203–1214. DOI: 10.1080/17512433.2017.1371591.Enioutina E. Yu., Teng L., Fateeva T. V., Brown J. C. S., Job K. M., Bortnikova V. V., Krepkova L. V., Gubarev M. I., Sherwin C. M. T. Phytotherapy as an alternative to conventional antimicrobials: combating microbial resistance. Expert Review of Clinical Pharmacology. 2017;10(11):1203–1214. DOI: 10.1080/17512433.2017.1371591.Abers M., Schroeder S., Goelz L., Sulser A., St Rose T., Puchalski K., Langland J. Antimicrobial activity of the volatile substances from essential oils. BMC Complementary Medicine and Therapies. 2021;21(1):124. DOI: 10.1186/s12906-021-03285-3.Abers M., Schroeder S., Goelz L., Sulser A., St Rose T., Puchalski K., Langland J. Antimicrobial activity of the volatile substances from essential oils. BMC Complementary Medicine and Therapies. 2021;21(1):124. DOI: 10.1186/s12906-021-03285-3.Jain S., Arora P., Nainwal L. M. Essential Oils as Potential Source of Anti-dandruff Agents: A Review. Combinatorial Chemistry & High Throughput Screening. 2022;25(9):1411–1426. DOI: 10.2174/1386207324666210712094148.Jain S., Arora P., Nainwal L. M. Essential Oils as Potential Source of Anti-dandruff Agents: A Review. Combinatorial Chemistry & High Throughput Screening. 2022;25(9):1411–1426. DOI: 10.2174/1386207324666210712094148.Mączka W., Duda-Madej A., Górny A., Grabarczyk M., Wińska K. Can eucalyptol replace antibiotics? Molecules. 2021;26(16):4933. DOI: 10.3390/molecules26164933.Mączka W., Duda-Madej A., Górny A., Grabarczyk M., Wińska K. Can eucalyptol replace antibiotics? Molecules. 2021;26(16):4933. DOI: 10.3390/molecules26164933.Mustarichie R., Rostinawati T., Pitaloka D. A. E., Saptarini N. M., Iskandar Y. Herbal Therapy for the Treatment of Seborrhea Dermatitis. Clinical, Cosmetic and Investigational Dermatology. 2022;2022(15):2391–2405. DOI: 10.2147/CCID.S376700.Mustarichie R., Rostinawati T., Pitaloka D. A. E., Saptarini N. M., Iskandar Y. Herbal Therapy for the Treatment of Seborrhea Dermatitis. Clinical, Cosmetic and Investigational Dermatology. 2022;2022(15):2391–2405. DOI: 10.2147/CCID.S376700.Ayatollahi A., Firooz A., Lotfali E., Mojab F., Fattahi M. Herbal Therapy for the Management of Seborrheic Dermatitis: A Narrative Review. Recent Advances in Anti-Infective Drug Discovery. 2021;16(3):209–226. DOI: 10.2174/2772434416666211029113213.Ayatollahi A., Firooz A., Lotfali E., Mojab F., Fattahi M. Herbal Therapy for the Management of Seborrheic Dermatitis: A Narrative Review. Recent Advances in Anti-Infective Drug Discovery. 2021;16(3):209–226. DOI: 10.2174/2772434416666211029113213.Brophy J. J., Davies N. W., Southwell I. A., Stiff I. A., Williams L. R. Gas chromatographic quality control for oil of Melaleuca terpinen-4-ol type (Australian tea tree). Journal of Agricultural and Food Chemistry. 1989;37:1330–1335.Brophy J. J., Davies N. W., Southwell I. A., Stiff I. A., Williams L. R. Gas chromatographic quality control for oil of Melaleuca terpinen-4-ol type (Australian tea tree). Journal of Agricultural and Food Chemistry. 1989;37:1330–1335.Carson C. F., Hammer K. A., Riley T. V. Melaleuca alternifolia (Tea Tree) Oil: a review of antimicrobial and other medicinal properties. Clinical Microbiology Reviews. 2006;19(1):50–62. DOI: 10.1128/CMR.19.1.50-62.2006.Carson C. F., Hammer K. A., Riley T. V. Melaleuca alternifolia (Tea Tree) Oil: a review of antimicrobial and other medicinal properties. Clinical Microbiology Reviews. 2006;19(1):50–62. DOI: 10.1128/CMR.19.1.50-62.2006.Hammer K. A., Carson C. F., Riley T. V. Antifungal activity of the components of Melaleuca alternifolia (tea tree) oil. Journal of Applied Microbiology. 2003;95(4):853–860. DOI: 10.1046/j.1365-2672.2003.02059.x.Hammer K. A., Carson C. F., Riley T. V. Antifungal activity of the components of Melaleuca alternifolia (tea tree) oil. Journal of Applied Microbiology. 2003;95(4):853–860. DOI: 10.1046/j.1365-2672.2003.02059.x.Brand C., Ferrante A., Prager R. H., Riley T. V., Carson C. F., Finlay-Jones J. J., Hart P. H. The water-soluble components of the essential oil of Melaleuca alternifolia (tea tree oil) suppress the production of superoxide by human monocytes, but not neutrophils, activated in vitro. Inflammation Research. 2001;50(4):213–219. DOI: 10.1007/s000110050746.Brand C., Ferrante A., Prager R. H., Riley T. V., Carson C. F., Finlay-Jones J. J., Hart P. H. The water-soluble components of the essential oil of Melaleuca alternifolia (tea tree oil) suppress the production of superoxide by human monocytes, but not neutrophils, activated in vitro. Inflammation Research. 2001;50(4):213–219. DOI: 10.1007/s000110050746.Waldroup W., Scheinfeld N. Medicated shampoos for the treatment of seborrheic dermatitis. Journal of Drugs in Dermatology. 2008;7(7):699–703.Waldroup W., Scheinfeld N. Medicated shampoos for the treatment of seborrheic dermatitis. Journal of Drugs in Dermatology. 2008;7(7):699–703.Satchell A. C., Saurajen A., Bell C., Barnetson R. S. Treatment of dandruff with 5% tea tree oil shampoo. Journal of the American Academy of Dermatology. 2002;47(6):852–855. DOI: 10.1067/mjd.2002.122734.Satchell A. C., Saurajen A., Bell C., Barnetson R. S. Treatment of dandruff with 5% tea tree oil shampoo. Journal of the American Academy of Dermatology. 2002;47(6):852–855. DOI: 10.1067/mjd.2002.122734.Dhakad A. K., Pandey V. V., Beg S., Rawat J. M., Singh A. Biological, medicinal and toxicological significance of Eucalyptus leaf essential oil: a review. Journal of the Science of Food and Agriculture. 2017;98(3):833–848. DOI: 10.1002/jsfa.8600.Dhakad A. K., Pandey V. V., Beg S., Rawat J. M., Singh A. Biological, medicinal and toxicological significance of Eucalyptus leaf essential oil: a review. Journal of the Science of Food and Agriculture. 2017;98(3):833–848. DOI: 10.1002/jsfa.8600.Sikkema J., de Bont J. A., Poolman B. Interactions of cyclic hydrocarbons with biological membranes. Journal of Biological Chemistry. 1994;269(11):8022–8028.Sikkema J., de Bont J. A., Poolman B. Interactions of cyclic hydrocarbons with biological membranes. Journal of Biological Chemistry. 1994;269(11):8022–8028.Zengin H., Baysal A. H. Antibacterial and antioxidant activity of essential oil terpenes against pathogenic and spoilage-forming bacteria and cell structure-activity relationships evaluated by SEM microscopy. Molecules. 2014;19(11):17773–177798. DOI: 10.3390/molecules191117773.Zengin H., Baysal A. H. Antibacterial and antioxidant activity of essential oil terpenes against pathogenic and spoilage-forming bacteria and cell structure-activity relationships evaluated by SEM microscopy. Molecules. 2014;19(11):17773–177798. DOI: 10.3390/molecules191117773.Kamatou G. P. P., Viljoen A. M. A review of the application and pharmacological properties of α-bisabolol and α-bisabolol-rich oils. Journal of the American Oil Chemists' Society. 2010;87(1):1–7.Kamatou G. P. P., Viljoen A. M. A review of the application and pharmacological properties of α-bisabolol and α-bisabolol-rich oils. Journal of the American Oil Chemists' Society. 2010;87(1):1–7.De Lucca A. J., Pauli A., Schilcher H., Sien T., Bhatnagar D., Walsh T. J. Fungicidal and Bactericidal Properties of Bisabolol and Dragosantol. Journal of Essential Oil Research. 2011;23(3):47–54.De Lucca A. J., Pauli A., Schilcher H., Sien T., Bhatnagar D., Walsh T. J. Fungicidal and Bactericidal Properties of Bisabolol and Dragosantol. Journal of Essential Oil Research. 2011;23(3):47–54.Filatov V. A., Kulyak O. Yu., Kalenikova E. I. Chemical Composition and Antimicrobial Potential of a Plant-Based Substance for the Treatment of Seborrheic Dermatitis. Pharmaceuticals. 2023;16(3):328. DOI: 10.3390/ph16030328.Filatov V. A., Kulyak O. Yu., Kalenikova E. I. Chemical Composition and Antimicrobial Potential of a Plant-Based Substance for the Treatment of Seborrheic Dermatitis. Pharmaceuticals. 2023;16(3):328. DOI: 10.3390/ph16030328.Filatov V. A., Kulyak O. Yu., Kalenikova E. I. In vitro and in vivo antimicrobial activity of an active plant-based quadrocomplex for skin hygiene. Journal of Pharmacy & Pharmacognosy Research. 2022;10(5):905–921.Filatov V. A., Kulyak O. Yu., Kalenikova E. I. In vitro and in vivo antimicrobial activity of an active plant-based quadrocomplex for skin hygiene. Journal of Pharmacy & Pharmacognosy Research. 2022;10(5):905–921.Filatov V. A., Ilin E. A., Kulyak O. Yu., Kalenikova E. I. Development and Validation of a Gas Chromatography–Mass Spectrometry Method for the Analysis of the Novel Plant-Based Substance with Antimicrobial Activity. Antibiotics. 2023;12(10):1558. DOI: 10.3390/antibiotics12101558.Filatov V. A., Ilin E. A., Kulyak O. Yu., Kalenikova E. I. Development and Validation of a Gas Chromatography–Mass Spectrometry Method for the Analysis of the Novel Plant-Based Substance with Antimicrobial Activity. Antibiotics. 2023;12(10):1558. DOI: 10.3390/antibiotics12101558.Ionov N., Druzhilovskiy D., Filimonov D., Poroikov V. Phyto4Health: Database of Phytocomponents from Russian Pharmacopoeia Plants. Journal of Chemical Information and Modeling. 2023;63(7):1847–1851. DOI: 10.1021/acs.jcim.2c01567.Ionov N., Druzhilovskiy D., Filimonov D., Poroikov V. Phyto4Health: Database of Phytocomponents from Russian Pharmacopoeia Plants. Journal of Chemical Information and Modeling. 2023;63(7):1847–1851. DOI: 10.1021/acs.jcim.2c01567.Bellio P., Fagnani L., Nazzicone L., Celenza G. New and simplified method for drug combination studies by checkerboard assay. MethodsX. 2021;8:101543. DOI: 10.1016/j.mex.2021.101543.Bellio P., Fagnani L., Nazzicone L., Celenza G. New and simplified method for drug combination studies by checkerboard assay. MethodsX. 2021;8:101543. DOI: 10.1016/j.mex.2021.101543.Odds F. C. Synergy, antagonism, and what the chequerboard puts between them. Journal of Antimicrobial Chemotherapy. 2003;52(1):1. DOI: 10.1093/jac/dkg301.Odds F. C. Synergy, antagonism, and what the chequerboard puts between them. Journal of Antimicrobial Chemotherapy. 2003;52(1):1. DOI: 10.1093/jac/dkg301.Piovan A., Caniato R., Brun P., Costa V. D., Filippini R. Rapid and feasible TLC screening of tea tree oil commercial samples. Journal of Pharmacognosy and Phytochemistry. 2021;10(1):175–180. DOI: 10.22271/phyto.2021.v10.i1c.13304.Piovan A., Caniato R., Brun P., Costa V. D., Filippini R. Rapid and feasible TLC screening of tea tree oil commercial samples. Journal of Pharmacognosy and Phytochemistry. 2021;10(1):175–180. DOI: 10.22271/phyto.2021.v10.i1c.13304.Shaha A., Salunkhe V. R. Development and validation of a high performance thin layer chromatographic method for determination of 1,8-Cineole in Callistemon Citrinus. Pharmacognosy Research. 2014;6(2):143–147.Shaha A., Salunkhe V. R. Development and validation of a high performance thin layer chromatographic method for determination of 1,8-Cineole in Callistemon Citrinus. Pharmacognosy Research. 2014;6(2):143–147.Hendry E. R., Worthington T., Conway B. R., Lambert P. A. Antimicrobial efficacy of eucalyptus oil and 1,8-cineole alone and in combination with chlorhexidine digluconate against microorganisms grown in planktonic and biofilm cultures. Journal of Antimicrobial Chemotherapy. 2009;64(6):1219–1225. DOI: 10.1093/jac/dkp362.Hendry E. R., Worthington T., Conway B. R., Lambert P. A. Antimicrobial efficacy of eucalyptus oil and 1,8-cineole alone and in combination with chlorhexidine digluconate against microorganisms grown in planktonic and biofilm cultures. Journal of Antimicrobial Chemotherapy. 2009;64(6):1219–1225. DOI: 10.1093/jac/dkp362.Filatov V. A., Kulyak O. Yu., Kalenikova E. I. The Development of Medical Shampoo with a Plant-Based Substance for the Treatment of Seborrheic Dermatitis. Medical Sciences Forum. 2023;21(1):51. DOI: 10.3390/ECB2023-14084.Filatov V. A., Kulyak O. Yu., Kalenikova E. I. The Development of Medical Shampoo with a Plant-Based Substance for the Treatment of Seborrheic Dermatitis. Medical Sciences Forum. 2023;21(1):51. DOI: 10.3390/ECB2023-14084.

The authors declare that there are no conflicts of interest present.