Planta Med 2023; 89(08): 790-799
DOI: 10.1055/a-1828-2479
Biological and Pharmacological Activity
Original Papers

In Vitro and In Silico Evaluation of ACE2 and LOX Inhibitory Activity of Origanum Essential Oils and Carvacrol[ # ]

1   Department of Pharmacognosy, Faculty of Pharmacy, Anadolu University, Eskişehir, Turkey
2   Faculty of Pharmacy, Eastern Mediterranean University, Famagusta, N. Cyprus, Mersin 10, Turkey
,
3   Department of Medical Biochemistry, Faculty of Medicine, Girne American University, Kyrenia, Mersin 10, Turkey
,
4   Department of Pharmacognosy, School of Pharmacy, Istanbul Medipol University, Beykoz, Istanbul, Turkey
5   Graduate School of Health Sciences, Anadolu University, Eskişehir, Turkey
,
6   Department of Pharmaceutical Microbiology, School of Pharmacy, Istanbul Medipol University, Beykoz, Istanbul, Turkey
7   Institute of Sciences, Istanbul University, Istanbul, Turkey
,
2   Faculty of Pharmacy, Eastern Mediterranean University, Famagusta, N. Cyprus, Mersin 10, Turkey
,
1   Department of Pharmacognosy, Faculty of Pharmacy, Anadolu University, Eskişehir, Turkey
,
2   Faculty of Pharmacy, Eastern Mediterranean University, Famagusta, N. Cyprus, Mersin 10, Turkey
,
8   Department of Pharmacognosy, Faculty of Pharmacy, Near East University, Nicosia, N. Cyprus, Mersin 10, Turkey
› Author Affiliations
Supported by: Anadolu University Scientific Research Projects Commission BAP 2005S058

Abstract

Origanum spp. are used both for culinary purposes and for their biological activities. In this study, commercial Origanum majorana, Origanum minutiflorum, Origanum vulgare, and Origanum onites essential oils and their prominent constituent carvacrol were evaluated for their in vitro and in silico angiotensin-converting enzyme 2 and lipoxygenase enzyme inhibitory potentials. The essential oils were analysed by gas chromatography-flame ionisation detection and gas chromatography-mass spectrometry, where carvacrol was identified as the major component (62 – 81%), confirming the quality. In vitro enzyme inhibition assays were conducted both with the essential oils (20 µg/mL) and with carvacrol (5 µg/mL). The comparative values of angiotensin-converting enzyme 2 percent inhibition for O. majorana, O. minutiflorum, O. vulgare, and O. onites essential oils were determined as 85.5, 79.1, 74.3, and 42.8%, respectively. As a result of the enzyme assays, carvacrol showed 90.7% in vitro angiotensin-converting enzyme 2 inhibitory activity. The in vitro lipoxygenase inhibition of the essential oils (in the same order) was 89.4, 78.9, 81.1, and 73.5%, respectively, where carvacrol showed 74.8% inhibition. In addition, protein–ligand docking and interaction profiling was used to gain structural and mechanistic insights into the angiotensin-converting enzyme 2 and lipoxygenase inhibitory potentials of major Origanum essential oil constituents. The in silico findings agreed with the significant enzyme inhibition activity observed in vitro. Further in vivo studies are suggested to confirm the safety and efficacy of the oils.

# Dedicated to Professor Dr. Gerhard Franz on the occasion of his 85th birthday.


Supporting Information



Publication History

Received: 01 December 2021

Accepted after revision: 08 April 2022

Accepted Manuscript online:
19 April 2022

Article published online:
28 September 2022

© 2022. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 
  • References

  • 1 Baser KHC, Özek T, Tümen G, Sezik E. Composition of the essential oils of turkish Origanum species with commercial importance. J Essent Oil Res 1993; 5: 619-623
  • 2 García-Beltrán JM, Esteban MA. Properties and applications of plants of Origanum sp. Genus. SM J Biol 2016; 2: 1006
  • 3 Taleb MH, Abdeltawab NF, Shamma RN, Abdelgayed SS, Mohamed SS, Farag MA, Ramadan MA. Origanum vulgare L. essential oil as a potential anti-acne topical nanoemulsion–in vitro and in vivo study. Molecules 2018; 23: 2164
  • 4 Costa MF, Durço AO, Rabelo TK, Barreto RSS, Guimaraes AG. Effects of carvacrol, thymol and essential oils containing such monoterpenes on wound healing: A systematic review. J Pharm Pharmacol 2019; 71: 141-155
  • 5 Avola R, Granata G, Geraci C, Napoli E, Graziano ACE, Cardile V. Oregano (Origanum vulgare L.) essential oil provides anti-inflammatory activity and facilitates wound healing in a human keratinocytes cell model. Food Chem Toxicol 2020; 144: 111586
  • 6 Samavati L, Uhal BD. ACE2, much more than just a receptor for SARS-COV-2. Front Cell Infect Microbiol 2020; 10: 317
  • 7 Letko M, Marzi A, Munster V. Functional assessment of cell entry and receptor usage for SARS-Cov-2 and other lineage B betacoronaviruses. Nat Microbiol 2020; 5: 562-569
  • 8 Teralı K, Baddal B, Gülcan HO. Prioritizing potential ACE2 inhibitors in the COVID-19 pandemic: Insights from a molecular mechanics-assisted structure-based virtual screening experiment. J Mol Graph Model 2020; 100: 107697
  • 9 Archambault AS, Zaid Y, Rakotoarivelo V, Turcotte C, Tore E, Dubuc I, Flamand L. High levels of eicosanoids and docosanoids in the lungs of intubated COVID-19 patients. FASEB J 2021; 35: e21666
  • 10 Snodgrass RG, Brüne B. Regulation and functions of 15-lipoxygenases in human macrophages. Front Pharmacol 2019; 10: 719
  • 11 Erdogan A, Ozkan A. Investigatıon of antioxidative, cytotoxic, membrane-damaging and membrane-protective effects of the essential oil of Origanum majorana and its oxygenated monoterpene component linalool in human-derived Hep G2 cell line. Iran J Pharm Res 2017; 16: 24
  • 12 Sokmen A, Abdel-Baki AAS, Al-Malki ES, Al-Quarishy S, Abdel-Haleem HM. Constituents of essential oil of Origanum minutiflorum and its in vitro antioxidant, scolicidal and anticancer activities. J King Saud Univ Sci 2020; 32: 2377-2382
  • 13 Faleiro L, Miguel G, Gomes S, Costa L, Venancio F, Teixeira A, Pedro LG. Antibacterial and antioxidant activities of essential oils isolated from Thymbra capitata L. (Cav.) and Origanum vulgare L. J Agric Food Chem 2005; 53: 8162-8168
  • 14 Ramírez D, Caballero J. Is it reliable to take the molecular docking top scoring position as the best solution without considering available structural data?. Molecules 2018; 23: 1038
  • 15 Towler P, Staker B, Prasad SG, Menon S, Tang J, Parsons T, Pantoliano MW. ACE2 X-ray structures reveal a large hinge-bending motion important for inhibitor binding and catalysis. J Biol Chem 2004; 279: 17996-18007
  • 16 Sarac N, Ugur A. The in vitro antimicrobial activities of the essential oils of some Lamiaceae species from Turkey. J Med Food 2009; 12: 902-907
  • 17 Ozel MZ, Kaymaz H. Superheated water extraction, steam distillation and Soxhlet extraction of essential oils of Origanum onites . Anal Bioanal Chem 2004; 379: 1127-1133
  • 18 Blank DE, Oliveira Hübner S, Alves GH, Cardoso CAL, Freitag RA, Cleff MB. Chemical composition and antiviral effect of extracts of Origanum vulgare . Adv Biosci Biotechnol 2019; 10: 188-196
  • 19 Khanavi M, Norouzi M, Tabatabaee H, Noudeh AS, Safavi SB, Shafiee A. Chemical compositions and antiviral effects of the essential oil of Zataria multiflora Boiss. and Origanum majorana L. J Med Plants 2010; 9: 128-137
  • 20 Games E, Guerreiro M, Santana FR, Pinheiro NM, De Oliveira DA, Lopes FD, Prado CM. Structurally related monoterpenes p-cymene, carvacrol and thymol isolated from essential oil from leaves of Lippia sidoides Cham. (Verbenaceae) protect mice against elastase-induced emphysema. Molecules 2016; 21: 1390
  • 21 Mediouni S, Jablonski JA, Tsuda S, Barsamia A, Kessing C, Richard A, Valente ST. Oregano oil and its principal component, carvacrol, inhibit HIV-1 fusion into target cells. J Virol 2020; 94: e00147-20
  • 22 Pilau MR, Alves SH, Weiblen R, Arenhart S, Cueto AP, Lovato LT. Antiviral activity of the Lippia graveolens (Mexican oregano) essential oil and its main compound carvacrol against human and animal viruses. Brazilian J Microbiol 2011; 42: 1616-1624
  • 23 Javed H, Meeran MFN, Jha NK, Ojha S. Carvacrol, a plant metabolite targeting viral protease (Mpro) and ACE2 in host cells can be a possible candidate for COVID-19. Front Plant Sci 2021; 11: 2237
  • 24 Asif M, Saleem M, Saadullah M, Yaseen HS, Al-Zarzour R. COVID-19 and therapy with essential oils having antiviral, anti-inflammatory, and immunomodulatory properties. Inflammopharmacology 2020; 28: 1153-1161
  • 25 Kulkarni SA, Nagarajan SK, Ramesh V, Palaniyandi V, Selvam SP, Madhavan T. Computational evaluation of major components from plant essential oils as potent inhibitors of SARS-CoV-2 spike protein. J Mol Struct 2020; 1221: 128823
  • 26 da Silva JKR, Figueiredo PLB, Byler KG, Setzer WN. Essential oils as antiviral agents. Potential of essential oils to treat SARS-CoV-2 infection: An in-silico investigation. Int J Mol Sci 2020; 21: 3426
  • 27 Gilbert NC, Gerstmeier J, Schexnaydre EE, Börner F, Garscha U, Neau DB, Newcomer ME. Structural and mechanistic insights into 5-lipoxygenase inhibition by natural products. Nat Chem Biol 2020; 16: 783-790
  • 28 Brash AR. Lipoxygenases: Occurrence, functions, catalysis, and acquisition of substrate. J Biol Chem 1999; 274: 23679-23682
  • 29 Somvanshi RK, Singh AK, Saxena M, Mishra B, Dey S. Development of novel peptide inhibitor of Lipoxygenase based on biochemical and BIAcore evidences. Biochim Biophys Acta 2008; 1784: 1812-1817
  • 30 Tsolaki E, Eleftheriou P, Kartsev V, Geronikaki A, Saxena AK. Application of docking analysis in the prediction and biological evaluation of the lipoxygenase inhibitory action of thiazolyl derivatives of mycophenolic acid. Molecules 2018; 23: 1621
  • 31 Taraporewala IB, Kauffman JM. Synthesis and structure-activity relationships of anti-inflammatory 9,10-dihydro-9-oxo-2-acridine-alkanoic acids and 4-(2-carboxyphenyl)aminobenzenealkanoic Acids. J Pharm Sci 1990; 79: 173-178
  • 32 Bahmani M, Khaksarian M, Rafieian-Kopaei M, Abbasi N. Overview of the therapeutic effects of Origanum vulgare and Hypericum perforatum based on Iranʼs ethnopharmacological documents. J Clin Diag Res 2018; 12: FE01-FE04
  • 33 Landa P, Kokoska L, Pribylova M, Vanek T, Marsik P. In vitro anti-inflammatory activity of carvacrol: Inhibitory effect on COX-2 catalyzed prostaglandin E2 biosynthesis. Arch Pharmacal Res 2009; 32: 75-78
  • 34 Lima MDS, Quintans-Júnior LJ, De Santana WA, Kaneto CM, Soares MBP, Villareal CP. Anti-inflammatory effects of carvacrol: Evidence for a key role of interleukin-10. Eur J Pharmacol 2013; 699: 112-117
  • 35 Demirci F, Karadağ AE, Biltekin SN, Demirci B. In vitro ACE2 and 5-LOX inhibition of Rosmarinus officinalis L. essential oil and its major component 1, 8-cineole. Rec Nat Prod 2022; 16: 194-199
  • 36 Kim S, Chen J, Cheng T, Gindulyte A, He J, He S, Boltın EE. PubChem in 2021: New data content and improved web interfaces. Nucleic Acids Res 2021; 49: D1388-D1395
  • 37 Burley SK, Bhikadiya C, Bi C, Bittrich S, Chen L, Crichlow GV, Zhuravleva M. RCSB Protein Data Bank: Powerful new tools for exploring 3D structures of biological macromolecules for basic and applied research and education fundamental biology, biomedicine, biotechnology, bioengineering and energy sciences. Nucleic Acids Res 2021; 49: D437-D451
  • 38 Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblat DM, Meng EC, Ferrin TE. UCSF Chimera–a visualization system for exploratory research and analysis. J Comput Chem 2004; 25: 1605-1612
  • 39 Shapovalov MV, Dunbrack RL. A smoothed backbone-dependent rotamer library for proteins derived from adaptive kernel density estimates and regressions. Structure 2011; 19: 844-858
  • 40 Schellhammer I, Rarey M. TrixX: structure-based molecule indexing for large-scale virtual screening in sublinear time. J Comput Aided Mol Des 2007; 21: 223-238
  • 41 Henzler AM, Urbaczek S, Hilbig M, Rarey M. An integrated approach to knowledge-driven structure-based virtual screening. J Comput Aided Mol Des 2014; 28: 927-939
  • 42 Flachsenberg F, Meyder A, Sommer K, Penner P, Rarey M. A consistent scheme for gradient-based optimization of protein–ligand poses. J Chem Inf Model 2020; 60: 6502-6522
  • 43 Bietz S, Urbaczek S, Schulz B, Rarey M. Protoss: a holistic approach to predict tautomers and protonation states in protein-ligand complexes. J Cheminform 2014; 6: 1-12
  • 44 Bateman A. UniProt: A worldwide hub of protein knowledge. Nucleic Acids Res 2019; 47: D506-D515
  • 45 Di Tommaso P, Moretti S, Xenarios I, Orobitg M, Montanyola A, Chang JM, Notredame J. T-Coffee: A web server for the multiple sequence alignment of protein and rna sequences using structural information and homology extension. Nucleic Acids Res 2011; 39: W13-W17
  • 46 Robert X, Gouet P. Deciphering key features in protein structures with the new ENDscript server. Nucleic Acids Res 2014; 42: W320-W324
  • 47 Demirci F, Karadağ AE, Biltekin SN, Demirci B. In Vitro ACE2 and 5-LOX enzyme inhibition by menthol and three different mint essential oils. Nat Prod Commun 2021; 16: 1-5
  • 48 Baylac S, Racine P. Inhibition of 5-lipoxygenase by essential oils and other natural fragrant extracts. Int J Aromather 2003; 13: 138-142