In-silico Assessment of the effects of Hyptis suaveolens as a natural source of Acetylcholine esterase (AchE )- targeting insecticide

In-silico Assessment of the effects of Hyptis suaveolens as a natural source of Acetylcholine esterase (AchE )- targeting insecticide | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Short Report In-silico Assessment of the effects of Hyptis suaveolens as a natural source of Acetylcholine esterase (AchE )- targeting insecticide Caroline Amuche Okoli, Titilayo Omolara Johnson This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6328543/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Developing effective insecticides is crucial to the global fight against vector-borne diseases, epidemics, and pandemics. Acetylcholinesterase (AChE), a serine hydrolase responsible for regulating acetylcholine levels in various organisms, is one of the most common targets of synthetic insecticides such as organophosphates and carbamates. However, widespread exposure to these chemicals has led to a significant decline in insect sensitivity, necessitating higher effective doses and consequently increasing human exposure to their toxic effects. As a result, the search for safer and more effective insecticides derived from natural sources has gained significant attention. This study aimed to computationally evaluate the AChE inhibitory potential of bioactive compounds from Hyptis suaveolens (L.) Poit, a medicinal plant widely used in traditional medicine across tropical regions. Using in silico approaches, 39 bioactive compounds were assessed through molecular docking, ligand and protein preparation, and toxicity prediction, utilizing computational tools such as Lotus, Chimera, PyRx, and ProTox3. Docking analysis revealed that Hyptis suaveolens compounds exhibited binding affinities ranging from − 10.5 to -3.8 kcal/mol against AChE. The standard reference ligand, 9-(3-phenylmethylamino)-1,2,3,4-tetrahydroacridine (C₂₀H₂₀N₂), had a docking score of -9.3 kcal/mol. Among the tested compounds, LTS0222826 exhibited the strongest binding affinity (-10.5 kcal/mol), followed by LTS0163613 (-9.7 kcal/mol), while LTS0238624 showed the weakest binding affinity (-3.8 kcal/mol). Additionally, LTS0163613 and LTS0107905 exhibited the least predicted toxicological effects on human physiological systems. Notably, both compounds showed no adverse effect on the respiratory system and other key biological functions when administered orally. This suggests their potential safety as natural insecticides and insect repellents, supporting their further exploration as alternatives to synthetic chemical insecticides. Acetylcholinesterase Binding affinity Hyptis suaveolens (L.) Poit Drosophila melanogaster Figures Figure 1 Figure 2 Figure 3 Figure 4 Background Acetylcholine (ACh) is one of the major neurotransmitters involved in neurotransmission in both vertebrates and invertebrates [ 1 ] and the regulation of its activity by AChE-catalysed hydrolysis is essential for the normal functioning and/or survival of these animals. Acetylcholinesterase is a cholinergic enzyme that catalyses the breakdown of acetylcholine to acetic acid and choline, thereby terminating nerve transmission at the synapses [ 2 ]. Inhibition of AchE activity forms the basis for the action of many insecticides including organophosphates and carbamates [ 3 ]. Inhibition of AChE by these insecticides causes the accumulation of acetylcholine in the synapses, making acetylcholine receptors to be permanently open and resulting in some toxicological outcomes [ 2 , 4 ]. The cholinergic stimulation induced by an AChE inhibitor (AChEI) may cause hyperactivity of excitable tissues, leading to fatal consequences like muscle paralysis, coma, and death. There is also, overwhelming production of oxygen/nitrogen derived free radicals leading to down-regulation of the antioxidant defense system [ 5 ] thereby exposing the insects to the toxic effects of the insecticides [ 5 , 6 ]. However, several insects and pests are becoming increasingly resistance to several insecticides including AChE-targeting insecticides. This is attributed to extensive use of chemical insecticides [ 7 – 9 ]. This has reduced the efficacy of these chemicals and increased human exposure to the toxic adverse effects of the chemicals [ 10 ]. These and other factors, such as environmental pollution raises the need for natural eco- friendly alternatives. Insect repellent plants are considered safe and environmentally friendly alternatives for controlling insects, which are vectors to several infectious human diseases. Some of these plants are sometimes used as ornamental plants around residential houses to repel insects and as indigenous traditional medicines in most of tropical areas [ 11 ] one of such plants is Hyptis suaveolens , commonly known as “wild hops”. It belongs to the family of Lamiaceae [ 12 ]. It is used traditionally in ethnomedicine for the treatment of various ailments including diabetes, fever, eczema, flatulence, cancers and headaches and there were no reports on adverse side effects on health. H. suaveolens (L) Poit has recently been shown to possess insecticidal and insect-repellent ability towards several insects, including mosquitoes [ 13 ], houseflies [ 14 ], sand flies [ 15 ] and fruit flies (Drosophila melanogaster) [ 16 ], hence, it might be a source of effective and non-toxic natural insecticides. Therefore, using Drosophila melanogaster as a model insect, we investigated the insecticidal and AChE inhibitory potentials of H. suaveolens (L) Poit and its bioactive compounds through in silico approach. Previously characterized bioactive compounds of H. suaveolens (L) Poit were screened for inhibitory activity against DmAChE through molecular docking analysis to identify possible AChEIs among the compounds. We also investigated the possible toxic effects of these compounds on humans. Materials and Methods In silico analysis of the AChE inhibitory potentials of bioactive compounds of Hyptis suaveolens (L) Poit. Ligand preparation Thirty-one bioactive compounds of Hyptis suaveolens (L) Poit were obtained from previous reports and Ethnobotanical Database [ 10 , 11 ][ 17 ]. The canonical SMILES of 10 compounds of Hyptis suaveolens (L) Poit from LOTUS database were converted to PDB format using Chimera 1.19 while the structure data file (SDF) format of standard ligand 9-(3-phenylmethylamino)-1,2,3,4-tetrahydroacridine (C₂₀H₂₀N₂), was obtained from Protein Database. The SDF format of compounds and standard ligand were uploaded to PyRx software and converted to PDBQT format using the OpenBabel plugin. The output files were minimized to obtain the minimum energy for the ligand docking. Protein preparation The crystal structure of Drosophila melanogaster Acetylcholinesterase (DmAChE) with PDB ID: 6XYY was obtained from the Protein Data Bank (PDB) repository. Using UCSF Chimera 1.19, the protein structure obtained was prepared via the protein preparation wizard panel. During the preparation processes, hydrogen atoms were added, bond orders were allocated, disulfide bonds were generated, the side chains and loops that are missing were replaced using prime. Water molecules located outside 3.0 Å of the heteroatoms were removed and the protein structure was minimized using OPLS3e and optimized using PROPKA [ 18 , 19 ]. The prepared proteins and ligands were uploaded to the PyRx software for molecular docking analysis. Molecular docking Molecular docking of the prepared ligands and proteins were performed using AutoDock vina in the PyRx workspace. Using blind docking, the ligands were docked into the binding pocket of the target protein. Grid space was set by targeting important amino acid residues selected through literature [ 20 ] and from UniProtKB Toxicity prediction The highest-ranking compound based on binding affinity against the target protein was selected for toxicity testing. The toxicity potential of the highest-ranking compounds based on binding affinity against DmAChE were selected to analyze their potential toxicity on human physiological systems using ProTox 3- prediction of toxicity software. Results Table 1 shows the bioactive compounds of Hyptis suaveolens (L) Poit obtained from Ethnobotanical Database. Thirty-nine (39) bioactive compounds of Hyptis suaveolens (L) Poit were virtually extracted from the plant using LOTUS database. Molecular docking analysis of bioactive compounds of Hyptis suaveolens (L) Poit against Drosophila melanogaster acetylcholine esterase (DmAChE) Hyptis suaveolens (L) Poit compounds exhibited various levels of binding affinities with Gibbs free energy (ΔG kcal/mol) ranked from the best target (– 10.5 kcal/mol) to worst (− 3.8 kcal/mol). Five (5) compounds of Hyptis suaveolens (L) Poit with the best docking scores against DmAChE. were selected with Gibbs free energy ranging from – 10.5 kcal/mol to -7.9 kcal/mol while the standard ligand (9-(3-phenylmethylamino)-1,2,3,4-tetrahydroacridine (C 20 H 20 N 2 ) showed a docking score of – 9.3 kcal/mol. Among the Hyptis suaveolens (L) Poit compounds, LTS0222826 scored highest with docking score of – 10.5 kcal/mol followed by LTS0163613 with a docking score of -9.7 kcal/mol while LTS0238624 scored lowest (-3.8 kcal/mol) (Table 1 ) Toxicity potentials of some compounds of Hyptis suaveolens (L) Poit Three compounds of Hyptis suaveolens (L) Poit (LTS0222826, LTS0107905 and LTS0163613) showed the least predicted toxicity effect on human physiological systems when consumed orally. LTS0107905 and LTS0163613 have no toxicity effect on the respiratory system (Table 2 and Figs. 2 – 4 ). Discussion Insecticide development is a very important aspect of the global fight against epidemics and pandemics. Insect-borne diseases like malaria and dengue hemorrhagic fever are among the deadliest infections worldwide [ 21 ] accounting for over 700 000 deaths yearly [ 22 ] and one of the ways of controlling them is through the use of insecticides. Acetylcholinesterase (AChE) is one of the most common targets of synthetic insecticides and pesticides, such as organophosphates and carbamate [ 23 ]. This enzyme is a serine hydrolase and is responsible for regulating the levels of acetylcholine in a variety of organisms, from mammals to insects [ 24 ]. Due to its extensive “attack”, there has beeen drastic reduction in the sensitivity of insects to organophosphate and carbamate insecticides. This has elevated the effective doses of these chemicals, leading to increased human exposure to their toxic adverse effects [ 10 ]. Hence the search for new, safe and effective insecticides of natural origin is currently being promoted as a good alternative [ 25 ]. This study has not only provided a scientific basis for the alternative use of Hyptis suaveolens (L) Poit as an insecticidal/insect-repellent plant, but it has further revealed its potential as a source of AChE-targeting agents and possible toxicological effects when consumed orally by humans In this study, during a search in the Protein Data Bank for Dm Acetylcholinesterase PDB_ID_6XYY, a structure a co-crystalized ligand (standard ligand (9-(3-phenylmethylamino)-1,2,3,4 - tetrahydroacridine ( C 20 H 20 N 2 ) to the protein was found. Also, to identify potential AChE inhibitors in Hyptis suaveolens (L) Poit, molecular docking analysis of some previously characterized bioactive compounds of the plant was conducted against DmAChE. The 39 compounds showed varying levels of binding affinities with docking scores ranging from – 10.5 to − 3.8 kcal/mol against the enzyme. The ten top-scoring Hyptis suaveolens (L) Poit, compounds were listed in Table 1 . The compound, LTS0222826 scored highest with docking score of – 10.5 kcal/mol followed by LTS0163613 with a docking score of - 9.7 kcal/mol. The 2D and 3D models of the protein-ligand interactions were not captured in this study. However, using PyRx virtual screening software showed that the 2 compounds were in close proximity or occupied the same binding pocket as the co-crystallized ligand (1qon) on the protein. This is in line with the report by Gaddaguti, et al. (2012) [ 26 ]. These interactions, therefore, make them potential inhibitors of DmAChE and possible contributors to the insecticidal or repellant activities of Hyptis suaveolens (L) Poit. This present study, therefore, identifies the compounds- LTS0222826 and LTS0163613 as the probable AChE-targeting agents and potential insecticides in Hyptis suaveolens (L) Poit. Though, additional computational and experimental studies need to be performed to further optimize and develop this hypothesis. Additionally, LTS0163613 and LTS0107905 exhibited the least predicted toxicological effects on human physiological systems. Notably, both compounds showed no adverse effect on the respiratory system and other key biological functions when administered orally. This suggests their potential safety as natural insecticides and insect repellents, supporting their further exploration as alternatives to synthetic chemical insecticides. Conclusion This present study, identifies LTS0222826 and LTS0163613 as the probable AChE-targeting agents and potential insecticides in Hyptis suaveolens (L) Poit. Also, LTS0163613 and LTS0107905 exhibited the least predicted toxicological effects on human physiological systems. Both showed no toxicological effect on the respiratory system among other systems and processes in the human body when consumed orally. This could affirm the safety of these compounds as natural insecticide and insect repellent, Notably, both compounds showed no adverse effect on the respiratory system and other key biological functions when administered orally. This suggests their potential safety as natural insecticides and insect repellents, supporting their further exploration as alternatives to synthetic chemical insecticides. Limitation of the study Additional computational and experimental studies are needed in order to further optimize and develop the hypothesis established in this study. Ethics declarations Not applicable Data availability Data will be made available on request. Declarations Ethics declarations Not applicable Data availability Data will be made available on request. Conflict of interest: There was no conflict of interest. Funding: This study was funded by the authors. Authors’ Contributions CAO conceived the study, participated in the in-silico analysis, interpretation and drafting of the original manuscript. TOJ mentored CAO and supervised the work, interpretated the results, reviewed and edited the manuscript. References Cupi ´ ´c D, Miladinovic S, Borozan S, Ivanovi´c. Involvement of cholinesterases in oxidative stress induced by chlorpyrifos in the brain of Japanese quail. Poult Sci. 2018;97:1564–71. Johnson TO, Abolaji AO, Omale S, Longdet YI, Kutshik JR, Oyetayo OB, et al. Benzo[a]pyrene and Benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide induced locomotor and reproductive senescence and altered biochemical parameters of oxidative damage in Canton-S Drosophila melanogaster. Toxicol Rep. 2021;8:571–80. Harel M, Kryger G, Rosenberry TL, Mallender WD, Lewis T, Fletcher RJ, et al. Three-dimensional structures of Drosophila melanogaster acetylcholinesterase and of its complexes with two potent inhibitors. Protein Sci. 2000;9:1063–72. Menozzi P, Shi MA, Lougarre A, Tang ZH, Fournier D. Mutations of acetylcholinesterase which confer insecticide resistance in Drosophila melanogaster populations. BMC Evol Biol. 2004;4:1–7. Milatovic D, Gupta RC, Aschner M. Anticholinesterase toxicity and oxidative stress, Sci. World J. 2006;6:295–310. https://doi.org/10.1100/tsw.2006.38 . Cupi ´ ´c D, Miladinovic S, Borozan S, Ivanovi´c. Involvement of cholinesterases in oxidative stress induced by chlorpyrifos in the brain of Japanese quail. Poult Sci. 2018;97:1564–71. Moustafa MAM, Awad M, Amer A, Hassan NN, Ibrahim E-DS, Ali HM, et al. Insecticidal activity of lemongrass essential oil as an eco-friendly agent against the black Cutworm Agrotis ipsilon (Lepidoptera: Noctuidae). J Adv Pharm Technol Res. 2021;2:737. https://doi.org/10.3390/insects12080737 . Shah G, Shri R, Panchal V, Sharma N, Singh B, Mann AS. Scientific basis for the therapeutic use of Cymbopogon citratus, stapf (Lemon grass). J Adv Pharm Technol Res. 2011;2:3–8. https://doi.org/10.4103/2231-4040.79796 . Shaikh MN, Suryawanshi YC, Mokat DN. Volatile profiling and essential oil yield of Cymbopogon citratus (DC.) stapf treated with Rhizosphere fungi and some important fertilizers. J Essent Oil Bear Plants. 2019;22:477–83. Trang A, Pb K. Physiology, Acetylcholinesterase. StatPearls [Internet]. StatPearls Publishing, Treasure Island (FL); 2021. Ellman GL, Courtney KD, Andres V, Featherstone RM. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol. 1961;7:88–90. Ashok Joseph R, Vijay V, Pandiyarajan, Petchimuthu K. Comparison Of Chemical Composition of the Essential Oil of Hyptis suaveolens (L.) Poit Leaves from Different Regions of Tamil Nadu. IJPSR, 2(11) 2822–4. Ojewumi ME, Banjo MG, Oresegun MO, Ogunbiyi TA, Ayoola AA, Awolu OO, et al. Analytical investigation of the extract of lemon grass leaves in repelling mosquito. Int J Pharm Sci Res. 2017;8:2048–55. https://doi.org/10.13040/IJPSR.0975-8232.8(5).1000-09 . Chintalchere JM, Dar MA, Raut KD, Pandit RS. Bioefficacy of lemongrass and tea tree essential oils against house fly, Musca domestica, Proc. Natl. Acad. Sci. India B Biol. Sci. 91 (2021) 307–318. https://doi.org/10.1007/s40011-020-01220- z Kimutai A, Ngeiywa M, Mulaa M, Njagi PGN, Ingonga J, Nyamwamu LB, et al. Repellent effects of the essential oils of Cymbopogon citratus and Tagetes minuta on the sandfly, Phlebotomus duboscqi. BMC Res Notes. 2017;10:1–9. https://doi.org/10.1186/s13104-017-2396-0 . Aljedani DM. Effects of some insecticides (Deltamethrin and malathion) and lemongrass oil on fruit fly (drosophila melanogaster). Pakistan J Biol Sci. 2021;24:477–91. Lotus- the. natural products occurrence database. https://lotus.naturalproducts.net/ Olsson MHM, SØndergaard CR, Rostkowski M, Jensen JH. PROPKA3: consistent treatment of internal and surface residues in empirical p K a prediction. J Chem Theor Comput. 2011;7:525–37. https://doi.org/10.1021/ct100578z . Iwaloye O, Elekofehinti OO, Momoh AI, Babatomiwa K, Ariyo EO. In silico molecular studies of natural compounds as possible anti-Alzheimer’s agents: ligand-based design. Netw Model Anal Heal Inf Bioinform. 2020;9:54. Bonikowski R, Switakowska P, Kula ´J. Synthesis, odour evaluation and antimicrobial activity of some geranyl acetone and nerolidol analogues. Flavour Fragr J. 2015;30:238–44. Manguin S, Mouchet J, Carnevale P. Main topics in entomology:Insects as disease vectors. RSC Green Chem 1–52 (2011). World Health Organisation. Vector-borne Diseases, 2020. Orhan I, Sener B, Choudhary MI, Khalid A. Acetylcholinesterase and butyrylcholinesterase inhibitory activity of some Turkish medicinal plants. J Ethnopharmacol. 2004;91:57–60. Dohi S, Terasaki M, Masakazu M. Acetylcholinesterase inhibitory activity and chemical composition of commercial essential oils. J Agric Food Chem. 2009;57:4313–8. Evans FE, Miller DW, Cairns T, Baddeley GV, Wenkert E. Structure analysis of proximadiol (Cryptomeridiol) by 13C NMR spectroscopy. Phytochemistry. 1982;21:937–8. Gaddaguti V, Mounika SJ, Sowjanya K, Rao T et al. Gc-Ms Analysis and In silico Molecular Docking Studies of Mosquito Repellent Compounds from Hyptis Suaveolens (L) Poit. Int J Bioassays 1(9) (2012). Tables Tables 1 and 2 are available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files Tables.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6328543","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Short Report","associatedPublications":[],"authors":[{"id":436413303,"identity":"42455a5a-1c68-43da-bd24-9a0ba7c7bdab","order_by":0,"name":"Caroline Amuche Okoli","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA3UlEQVRIie3RoQoCMRzH8Z8IWqZWRe4dDgZiEJ9l4+CewWAQBNNp9y3WxPaXgRb1HsDiNaPxmm6C2uZsgvumLXzY/hsQCv1iDBUSGABVCLuvTDwIDEm/JYB+7T+TFtNE53EuV3UURYlBpKihLy7Smc8Eie1JrqdIOEPKFTXTvovEOYtJTk5SaaRdc0OpiPViD3J8kE6Jmwc5ZJbQg7QZyBJ+ds6Sbe0sCTfEzBInfKlZzyXMiyWbohwPI5XvZVGOhtFil/Gr07xjwtwT9k9rbU9Sp+eq6ntKKBQK/Ud3JCpQ65USQh4AAAAASUVORK5CYII=","orcid":"","institution":"University of Jos","correspondingAuthor":true,"prefix":"","firstName":"Caroline","middleName":"Amuche","lastName":"Okoli","suffix":""},{"id":436413304,"identity":"f339be9d-31c9-4a9b-9624-068c528c8385","order_by":1,"name":"Titilayo Omolara Johnson","email":"","orcid":"","institution":"University of Jos","correspondingAuthor":false,"prefix":"","firstName":"Titilayo","middleName":"Omolara","lastName":"Johnson","suffix":""}],"badges":[],"createdAt":"2025-03-28 13:23:37","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6328543/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6328543/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":79655128,"identity":"03b0f0de-e2ec-4811-aebc-6994a7353f23","added_by":"auto","created_at":"2025-04-01 08:45:14","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":41965,"visible":true,"origin":"","legend":"\u003cp\u003eStructures of some compounds of Hyptis suaveolens (L.) Poit extracted from Lotus \u0026nbsp;\u0026nbsp;database\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6328543/v1/580882f1442820191b6300ce.png"},{"id":79655130,"identity":"f4624740-b19a-4ff9-9eb5-9a06908e5411","added_by":"auto","created_at":"2025-04-01 08:45:14","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":151101,"visible":true,"origin":"","legend":"\u003cp\u003eToxicity radar chart of LTS0222826\u003c/p\u003e\n\u003cp\u003e*\u003cstrong\u003e \u003c/strong\u003eThe toxicity radar chart is intended to quickly illustrate the confidence of positive toxicity results compared to the average of its class.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6328543/v1/7a20e4267028162b491d5612.png"},{"id":79655131,"identity":"f50a4799-f840-440f-9c05-7e4b561b1e61","added_by":"auto","created_at":"2025-04-01 08:45:15","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":151573,"visible":true,"origin":"","legend":"\u003cp\u003eToxicity radar chart of LTS0107905\u003c/p\u003e\n\u003cp\u003e*\u003cstrong\u003e \u003c/strong\u003eThe toxicity radar chart is intended to quickly illustrate the confidence of positive toxicity results compared to the average of its class.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6328543/v1/730ce5ae3f79901ccde6eda7.png"},{"id":79655139,"identity":"c5ff1ace-cfa1-441b-b293-dc9b93dfdef2","added_by":"auto","created_at":"2025-04-01 08:45:15","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":151322,"visible":true,"origin":"","legend":"\u003cp\u003eToxicity radar chart of LTS0163613\u003c/p\u003e\n\u003cp\u003e*\u003cstrong\u003e \u003c/strong\u003eThe toxicity radar chart is intended to quickly illustrate the confidence of positive toxicity results compared to the average of its class.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6328543/v1/a53a4d366b5169cfb294fc3e.png"},{"id":80384214,"identity":"9e448095-1c13-488f-a11e-b864fb8207ff","added_by":"auto","created_at":"2025-04-11 09:47:06","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":933563,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6328543/v1/fc71401d-2891-4ed3-8eef-c7dd95e1be83.pdf"},{"id":79655126,"identity":"f873776f-2801-4b93-8bd0-cee9c82fd18b","added_by":"auto","created_at":"2025-04-01 08:45:14","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":21296,"visible":true,"origin":"","legend":"","description":"","filename":"Tables.docx","url":"https://assets-eu.researchsquare.com/files/rs-6328543/v1/15f448a9d45901b9fd085d65.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"In-silico Assessment of the effects of Hyptis suaveolens as a natural source of Acetylcholine esterase (AchE )- targeting insecticide","fulltext":[{"header":"Background","content":"\u003cp\u003eAcetylcholine (ACh) is one of the major neurotransmitters involved in neurotransmission in both vertebrates and invertebrates [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e] and the regulation of its activity by AChE-catalysed hydrolysis is essential for the normal functioning and/or survival of these animals. Acetylcholinesterase is a cholinergic enzyme that catalyses the breakdown of acetylcholine to acetic acid and choline, thereby terminating nerve transmission at the synapses [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Inhibition of AchE activity forms the basis for the action of many insecticides including organophosphates and carbamates [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eInhibition of AChE by these insecticides causes the accumulation of acetylcholine in the synapses, making acetylcholine receptors to be permanently open and resulting in some toxicological outcomes [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. The cholinergic stimulation induced by an AChE inhibitor (AChEI) may cause hyperactivity of excitable tissues, leading to fatal consequences like muscle paralysis, coma, and death. There is also, overwhelming production of oxygen/nitrogen derived free radicals leading to down-regulation of the antioxidant defense system [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e] thereby exposing the insects to the toxic effects of the insecticides [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eHowever, several insects and pests are becoming increasingly resistance to several insecticides including AChE-targeting insecticides. This is attributed to extensive use of chemical insecticides [\u003cspan additionalcitationids=\"CR8\" citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. This has reduced the efficacy of these chemicals and increased human exposure to the toxic adverse effects of the chemicals [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. These and other factors, such as environmental pollution raises the need for natural eco- friendly alternatives. Insect repellent plants are considered safe and environmentally friendly alternatives for controlling insects, which are vectors to several infectious human diseases.\u003c/p\u003e \u003cp\u003eSome of these plants are sometimes used as ornamental plants around residential houses to repel insects and as indigenous traditional medicines in most of tropical areas [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e] one of such plants is \u003cem\u003eHyptis suaveolens\u003c/em\u003e, commonly known as \u0026ldquo;wild hops\u0026rdquo;. It belongs to the family of Lamiaceae [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. It is used traditionally in ethnomedicine for the treatment of various ailments including diabetes, fever, eczema, flatulence, cancers and headaches and there were no reports on adverse side effects on health. H. suaveolens (L) Poit has recently been shown to possess insecticidal and insect-repellent ability towards several insects, including mosquitoes [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e], houseflies [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], sand flies [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e] and fruit flies (Drosophila melanogaster) [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e], hence, it might be a source of effective and non-toxic natural insecticides.\u003c/p\u003e \u003cp\u003eTherefore, using Drosophila melanogaster as a model insect, we investigated the insecticidal and AChE inhibitory potentials of H. suaveolens (L) Poit and its bioactive compounds through in silico approach. Previously characterized bioactive compounds of H. suaveolens (L) Poit were screened for inhibitory activity against DmAChE through molecular docking analysis to identify possible AChEIs among the compounds. We also investigated the possible toxic effects of these compounds on humans.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003eIn silico analysis of the AChE inhibitory potentials of bioactive compounds of Hyptis suaveolens (L) Poit.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eLigand preparation\u003c/h2\u003e \u003cp\u003eThirty-one bioactive compounds of Hyptis suaveolens (L) Poit were obtained from previous reports and Ethnobotanical Database [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e][\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. The canonical SMILES of 10 compounds of Hyptis suaveolens (L) Poit from LOTUS database were converted to PDB format using Chimera 1.19 while the structure data file (SDF) format of standard ligand 9-(3-phenylmethylamino)-1,2,3,4-tetrahydroacridine (C₂₀H₂₀N₂), was obtained from Protein Database. The SDF format of compounds and standard ligand were uploaded to PyRx software and converted to PDBQT format using the OpenBabel plugin. The output files were minimized to obtain the minimum energy for the ligand docking.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eProtein preparation\u003c/h3\u003e\n\u003cp\u003eThe crystal structure of Drosophila melanogaster Acetylcholinesterase (DmAChE) with PDB ID: 6XYY was obtained from the Protein Data Bank (PDB) repository. Using UCSF Chimera 1.19, the protein structure obtained was prepared via the protein preparation wizard panel.\u003c/p\u003e \u003cp\u003eDuring the preparation processes, hydrogen atoms were added, bond orders were allocated, disulfide bonds were generated, the side chains and loops that are missing were replaced using prime. Water molecules located outside 3.0 \u0026Aring; of the heteroatoms were removed and the protein structure was minimized using OPLS3e and optimized using PROPKA [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. The prepared proteins and ligands were uploaded to the PyRx software for molecular docking analysis.\u003c/p\u003e\n\u003ch3\u003eMolecular docking\u003c/h3\u003e\n\u003cp\u003eMolecular docking of the prepared ligands and proteins were performed using AutoDock vina in the PyRx workspace. Using blind docking, the ligands were docked into the binding pocket of the target protein. Grid space was set by targeting important amino acid residues selected through literature [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e] and from UniProtKB\u003c/p\u003e\n\u003ch3\u003eToxicity prediction\u003c/h3\u003e\n\u003cp\u003eThe highest-ranking compound based on binding affinity against the target protein was selected for toxicity testing. The toxicity potential of the highest-ranking compounds based on binding affinity against DmAChE were selected to analyze their potential toxicity on human physiological systems using ProTox 3- prediction of toxicity software.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eTable \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e shows the bioactive compounds of Hyptis suaveolens (L) Poit obtained from Ethnobotanical Database. Thirty-nine (39) bioactive compounds of Hyptis suaveolens (L) Poit were virtually extracted from the plant using LOTUS database.\u003c/p\u003e\n\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n \u003cdiv id=\"Sec12\" class=\"Section3\"\u003e\n \u003cp\u003e\u003cstrong\u003eMolecular docking analysis of bioactive compounds of Hyptis suaveolens (L) Poit against Drosophila melanogaster acetylcholine esterase (DmAChE)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eHyptis suaveolens (L) Poit compounds exhibited various levels of binding affinities with Gibbs free energy (\u0026Delta;G kcal/mol) ranked from the best target (\u0026ndash; 10.5 kcal/mol) to worst (\u0026minus;\u0026thinsp;3.8 kcal/mol). Five (5) compounds of \u003cem\u003eHyptis suaveolens (L) Poit\u003c/em\u003e with the best docking scores against DmAChE. were selected with Gibbs free energy ranging from \u0026ndash; 10.5 kcal/mol to -7.9 kcal/mol while the standard ligand (9-(3-phenylmethylamino)-1,2,3,4-tetrahydroacridine (C\u003csub\u003e20\u003c/sub\u003e H\u003csub\u003e20\u003c/sub\u003e N\u003csub\u003e2\u003c/sub\u003e) showed a docking score of \u0026ndash; 9.3 kcal/mol. Among the \u003cem\u003eHyptis suaveolens (L) Poit\u003c/em\u003e compounds, LTS0222826 scored highest with docking score of \u0026ndash; 10.5 kcal/mol followed by LTS0163613 with a docking score of -9.7 kcal/mol while LTS0238624 scored lowest (-3.8 kcal/mol) (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e)\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n \u003ch2\u003eToxicity potentials of some compounds of Hyptis suaveolens (L) Poit\u003c/h2\u003e\n \u003cp\u003eThree compounds of Hyptis suaveolens (L) Poit (LTS0222826, LTS0107905 and LTS0163613) showed the least predicted toxicity effect on human physiological systems when consumed orally. LTS0107905 and LTS0163613 have no toxicity effect on the respiratory system (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e and Figs. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e\u0026ndash;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u0026nbsp;\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eInsecticide development is a very important aspect of the global fight against epidemics and pandemics. Insect-borne diseases like malaria and dengue hemorrhagic fever are among the deadliest infections worldwide [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e] accounting for over 700 000 deaths yearly [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e] and one of the ways of controlling them is through the use of insecticides. Acetylcholinesterase (AChE) is one of the most common targets of synthetic insecticides and pesticides, such as organophosphates and carbamate [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. This enzyme is a serine hydrolase and is responsible for regulating the levels of acetylcholine in a variety of organisms, from mammals to insects [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. Due to its extensive \u0026ldquo;attack\u0026rdquo;, there has beeen drastic reduction in the sensitivity of insects to organophosphate and carbamate insecticides. This has elevated the effective doses of these chemicals, leading to increased human exposure to their toxic adverse effects [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Hence the search for new, safe and effective insecticides of natural origin is currently being promoted as a good alternative [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThis study has not only provided a scientific basis for the alternative use of Hyptis suaveolens (L) Poit as an insecticidal/insect-repellent plant, but it has further revealed its potential as a source of AChE-targeting agents and possible toxicological effects when consumed orally by humans\u003c/p\u003e \u003cp\u003eIn this study, during a search in the Protein Data Bank for Dm Acetylcholinesterase PDB_ID_6XYY, a structure a co-crystalized ligand (standard ligand (9-(3-phenylmethylamino)-1,2,3,4\u003cb\u003e-\u003c/b\u003etetrahydroacridine \u003cb\u003e(\u003c/b\u003eC\u003csub\u003e20\u003c/sub\u003e H\u003csub\u003e20\u003c/sub\u003e N\u003csub\u003e2\u003c/sub\u003e) to the protein was found.\u003c/p\u003e \u003cp\u003eAlso, to identify potential AChE inhibitors in Hyptis suaveolens (L) Poit, molecular docking analysis of some previously characterized bioactive compounds of the plant was conducted against DmAChE. The 39 compounds showed varying levels of binding affinities with docking scores ranging from \u0026ndash; 10.5 to \u0026minus;\u0026thinsp;3.8 kcal/mol against the enzyme. The ten top-scoring Hyptis suaveolens (L) Poit, compounds were listed in Table \u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The compound, LTS0222826 scored highest with docking score of \u0026ndash; 10.5 kcal/mol followed by LTS0163613 with a docking score of \u003cb\u003e-\u003c/b\u003e9.7 kcal/mol. The 2D and 3D models of the protein-ligand interactions were not captured in this study. However, using PyRx virtual screening software showed that the 2 compounds were in close proximity or occupied the same binding pocket as the co-crystallized ligand (1qon) on the protein.\u003c/p\u003e \u003cp\u003eThis is in line with the report by Gaddaguti, et al. (2012) [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. These interactions, therefore, make them potential inhibitors of DmAChE and possible contributors to the insecticidal or repellant activities of Hyptis suaveolens (L) Poit. This present study, therefore, identifies the compounds- LTS0222826 and LTS0163613 as the probable AChE-targeting agents and potential insecticides in Hyptis suaveolens (L) Poit. Though, additional computational and experimental studies need to be performed to further optimize and develop this hypothesis.\u003c/p\u003e \u003cp\u003eAdditionally, LTS0163613 and LTS0107905 exhibited the least predicted toxicological effects on human physiological systems. Notably, both compounds showed no adverse effect on the respiratory system and other key biological functions when administered orally. This suggests their potential safety as natural insecticides and insect repellents, supporting their further exploration as alternatives to synthetic chemical insecticides.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis present study, identifies LTS0222826 and LTS0163613 as the probable AChE-targeting agents and potential insecticides in Hyptis suaveolens (L) Poit. Also, LTS0163613 and LTS0107905 exhibited the least predicted toxicological effects on human physiological systems. Both showed no toxicological effect on the respiratory system among other systems and processes in the human body when consumed orally. This could affirm the safety of these compounds as natural insecticide and insect repellent, Notably, both compounds showed no adverse effect on the respiratory system and other key biological functions when administered orally. This suggests their potential safety as natural insecticides and insect repellents, supporting their further exploration as alternatives to synthetic chemical insecticides.\u003c/p\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eLimitation of the study\u003c/h2\u003e \u003cp\u003eAdditional computational and experimental studies are needed in order to further optimize and develop the hypothesis established in this study.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eEthics declarations\u003c/h2\u003e \u003cp\u003eNot applicable\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eData availability\u003c/h2\u003e \u003cp\u003eData will be made available on request.\u003c/p\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics declarations\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData will be made available on request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThere was no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was funded by the authors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCAO conceived the study, participated in the in-silico analysis, interpretation and drafting of the original manuscript. TOJ mentored CAO and supervised the work, interpretated the results, reviewed and edited the manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eCupi \u0026acute; \u0026acute;c D, Miladinovic S, Borozan S, Ivanovi\u0026acute;c. Involvement of cholinesterases in oxidative stress induced by chlorpyrifos in the brain of Japanese quail. Poult Sci. 2018;97:1564\u0026ndash;71.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJohnson TO, Abolaji AO, Omale S, Longdet YI, Kutshik JR, Oyetayo OB, et al. Benzo[a]pyrene and Benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide induced locomotor and reproductive senescence and altered biochemical parameters of oxidative damage in Canton-S Drosophila melanogaster. Toxicol Rep. 2021;8:571\u0026ndash;80.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHarel M, Kryger G, Rosenberry TL, Mallender WD, Lewis T, Fletcher RJ, et al. Three-dimensional structures of Drosophila melanogaster acetylcholinesterase and of its complexes with two potent inhibitors. Protein Sci. 2000;9:1063\u0026ndash;72.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMenozzi P, Shi MA, Lougarre A, Tang ZH, Fournier D. Mutations of acetylcholinesterase which confer insecticide resistance in Drosophila melanogaster populations. BMC Evol Biol. 2004;4:1\u0026ndash;7.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMilatovic D, Gupta RC, Aschner M. Anticholinesterase toxicity and oxidative stress, Sci. World J. 2006;6:295\u0026ndash;310. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1100/tsw.2006.38\u003c/span\u003e\u003cspan address=\"10.1100/tsw.2006.38\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCupi \u0026acute; \u0026acute;c D, Miladinovic S, Borozan S, Ivanovi\u0026acute;c. Involvement of cholinesterases in oxidative stress induced by chlorpyrifos in the brain of Japanese quail. Poult Sci. 2018;97:1564\u0026ndash;71.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMoustafa MAM, Awad M, Amer A, Hassan NN, Ibrahim E-DS, Ali HM, et al. Insecticidal activity of lemongrass essential oil as an eco-friendly agent against the black Cutworm Agrotis ipsilon (Lepidoptera: Noctuidae). J Adv Pharm Technol Res. 2021;2:737. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/insects12080737\u003c/span\u003e\u003cspan address=\"10.3390/insects12080737\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShah G, Shri R, Panchal V, Sharma N, Singh B, Mann AS. Scientific basis for the therapeutic use of Cymbopogon citratus, stapf (Lemon grass). J Adv Pharm Technol Res. 2011;2:3\u0026ndash;8. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.4103/2231-4040.79796\u003c/span\u003e\u003cspan address=\"10.4103/2231-4040.79796\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShaikh MN, Suryawanshi YC, Mokat DN. Volatile profiling and essential oil yield of Cymbopogon citratus (DC.) stapf treated with Rhizosphere fungi and some important fertilizers. J Essent Oil Bear Plants. 2019;22:477\u0026ndash;83.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTrang A, Pb K. Physiology, Acetylcholinesterase. StatPearls [Internet]. StatPearls Publishing, Treasure Island (FL); 2021.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEllman GL, Courtney KD, Andres V, Featherstone RM. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol. 1961;7:88\u0026ndash;90.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAshok Joseph R, Vijay V, Pandiyarajan, Petchimuthu K. Comparison Of Chemical Composition of the Essential Oil of Hyptis suaveolens (L.) Poit Leaves from Different Regions of Tamil Nadu. IJPSR, 2(11) 2822\u0026ndash;4.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOjewumi ME, Banjo MG, Oresegun MO, Ogunbiyi TA, Ayoola AA, Awolu OO, et al. Analytical investigation of the extract of lemon grass leaves in repelling mosquito. Int J Pharm Sci Res. 2017;8:2048\u0026ndash;55. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.13040/IJPSR.0975-8232.8(5).1000-09\u003c/span\u003e\u003cspan address=\"10.13040/IJPSR.0975-8232.8(5).1000-09\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChintalchere JM, Dar MA, Raut KD, Pandit RS. Bioefficacy of lemongrass and tea tree essential oils against house fly, Musca domestica, Proc. Natl. Acad. Sci. India B Biol. Sci. 91 (2021) 307\u0026ndash;318. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s40011-020-01220- z\u003c/span\u003e\u003cspan address=\"10.1007/s40011-020-01220- z\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKimutai A, Ngeiywa M, Mulaa M, Njagi PGN, Ingonga J, Nyamwamu LB, et al. Repellent effects of the essential oils of Cymbopogon citratus and Tagetes minuta on the sandfly, Phlebotomus duboscqi. BMC Res Notes. 2017;10:1\u0026ndash;9. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s13104-017-2396-0\u003c/span\u003e\u003cspan address=\"10.1186/s13104-017-2396-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAljedani DM. Effects of some insecticides (Deltamethrin and malathion) and lemongrass oil on fruit fly (drosophila melanogaster). Pakistan J Biol Sci. 2021;24:477\u0026ndash;91.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLotus- the. natural products occurrence database. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://lotus.naturalproducts.net/\u003c/span\u003e\u003cspan address=\"https://lotus.naturalproducts.net/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOlsson MHM, S\u0026Oslash;ndergaard CR, Rostkowski M, Jensen JH. PROPKA3: consistent treatment of internal and surface residues in empirical p K a prediction. J Chem Theor Comput. 2011;7:525\u0026ndash;37. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1021/ct100578z\u003c/span\u003e\u003cspan address=\"10.1021/ct100578z\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIwaloye O, Elekofehinti OO, Momoh AI, Babatomiwa K, Ariyo EO. In silico molecular studies of natural compounds as possible anti-Alzheimer\u0026rsquo;s agents: ligand-based design. Netw Model Anal Heal Inf Bioinform. 2020;9:54.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBonikowski R, Switakowska P, Kula \u0026acute;J. Synthesis, odour evaluation and antimicrobial activity of some geranyl acetone and nerolidol analogues. Flavour Fragr J. 2015;30:238\u0026ndash;44.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eManguin S, Mouchet J, Carnevale P. Main topics in entomology:Insects as disease vectors. RSC Green Chem 1\u0026ndash;52 (2011).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWorld Health Organisation. Vector-borne Diseases, 2020.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOrhan I, Sener B, Choudhary MI, Khalid A. Acetylcholinesterase and butyrylcholinesterase inhibitory activity of some Turkish medicinal plants. J Ethnopharmacol. 2004;91:57\u0026ndash;60.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDohi S, Terasaki M, Masakazu M. Acetylcholinesterase inhibitory activity and chemical composition of commercial essential oils. J Agric Food Chem. 2009;57:4313\u0026ndash;8.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEvans FE, Miller DW, Cairns T, Baddeley GV, Wenkert E. Structure analysis of proximadiol (Cryptomeridiol) by 13C NMR spectroscopy. Phytochemistry. 1982;21:937\u0026ndash;8.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGaddaguti V, Mounika SJ, Sowjanya K, Rao T et al. Gc-Ms Analysis and In silico Molecular Docking Studies of Mosquito Repellent Compounds from Hyptis Suaveolens (L) Poit. Int J Bioassays 1(9) (2012).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003eTables 1 and 2 are available in the Supplementary Files section.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Acetylcholinesterase, Binding affinity, Hyptis suaveolens (L.) Poit, Drosophila melanogaster","lastPublishedDoi":"10.21203/rs.3.rs-6328543/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6328543/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eDeveloping effective insecticides is crucial to the global fight against vector-borne diseases, epidemics, and pandemics. Acetylcholinesterase (AChE), a serine hydrolase responsible for regulating acetylcholine levels in various organisms, is one of the most common targets of synthetic insecticides such as organophosphates and carbamates. However, widespread exposure to these chemicals has led to a significant decline in insect sensitivity, necessitating higher effective doses and consequently increasing human exposure to their toxic effects. As a result, the search for safer and more effective insecticides derived from natural sources has gained significant attention. This study aimed to computationally evaluate the AChE inhibitory potential of bioactive compounds from Hyptis suaveolens (L.) Poit, a medicinal plant widely used in traditional medicine across tropical regions. Using in silico approaches, 39 bioactive compounds were assessed through molecular docking, ligand and protein preparation, and toxicity prediction, utilizing computational tools such as Lotus, Chimera, PyRx, and ProTox3. Docking analysis revealed that Hyptis suaveolens compounds exhibited binding affinities ranging from \u0026minus;\u0026thinsp;10.5 to -3.8 kcal/mol against AChE. The standard reference ligand, 9-(3-phenylmethylamino)-1,2,3,4-tetrahydroacridine (C₂₀H₂₀N₂), had a docking score of -9.3 kcal/mol. Among the tested compounds, LTS0222826 exhibited the strongest binding affinity (-10.5 kcal/mol), followed by LTS0163613 (-9.7 kcal/mol), while LTS0238624 showed the weakest binding affinity (-3.8 kcal/mol). Additionally, LTS0163613 and LTS0107905 exhibited the least predicted toxicological effects on human physiological systems. Notably, both compounds showed no adverse effect on the respiratory system and other key biological functions when administered orally. This suggests their potential safety as natural insecticides and insect repellents, supporting their further exploration as alternatives to synthetic chemical insecticides.\u003c/p\u003e","manuscriptTitle":"In-silico Assessment of the effects of Hyptis suaveolens as a natural source of Acetylcholine esterase (AchE )- targeting insecticide","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-01 08:45:10","doi":"10.21203/rs.3.rs-6328543/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"798115b8-3c48-4cb8-8c2c-93ec3c97b690","owner":[],"postedDate":"April 1st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-04-11T09:38:58+00:00","versionOfRecord":[],"versionCreatedAt":"2025-04-01 08:45:10","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6328543","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6328543","identity":"rs-6328543","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}