Synthesis, Spectroscopic Characterization, and In Silico Evaluation of 3-Methyl-6-anilino-1,2,4-triazine as a Promising Antidiabetic Agent

Synthesis, Spectroscopic Characterization, and In Silico Evaluation of 3-Methyl-6-anilino-1,2,4-triazine as a Promising Antidiabetic Agent | 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 Research Article Synthesis, Spectroscopic Characterization, and In Silico Evaluation of 3-Methyl-6-anilino-1,2,4-triazine as a Promising Antidiabetic Agent Prafulkumar R Valand, Jayshreeben N Patel This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6651760/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 The triazine scaffold remains a cornerstone in the development of biologically active molecules due to its structural versatility and pharmacological potential. In this study, we report the synthesis of 3-methyl-6-anilino-1,2,4-triazine, a brown crystalline compound obtained through the condensation of 3-methyl-6-amino-1,2,4-triazine with aniline hydrochloride. Structural elucidation was carried out via IR, 1H NMR, 13C NMR, and mass spectrometry, confirming the proposed framework. The compound was subjected to SwissADME analysis, which indicated favorable pharmacokinetic properties, high gastrointestinal absorption, and compliance with Lipinski's rule. Molecular docking was performed against PPAR-γ (PDB ID: 3VI8), showing a binding energy of -7.6 kcal/mol and key hydrogen bonding interactions. These findings suggest that the compound could serve as a viable lead for the development of new antidiabetic agents. 1 2 4-triazine PPAR-γ molecular docking antidiabetic drug-likeness SwissADME Introduction Diabetes mellitus is a Pandemic disease that has struck each and every corner of the world. According to Indian Council of Medical Research-Indian Diabetes Study (ICMR), a national diabetes study, India Currently has an estimated 77 million people with diabetes, which makes it the second most affected in the world after China. This is set to increase to over 100 million by 2030( 1 ). The Prevalence of diabetes among adults has reached approximately 20% in Urban and approximately 10% in rural populations in India ( 2 ). Various classes of antidiabetic drugs including insulin and oral hypoglycemic agents (OHA) are currently used in the treatment of diabetes which acts by different mechanism to reduce blood glucose levels to maintain optimal glycemic control ( 3 , 4 ). The United Kingdom Prospective Diabetes Study showed intensive blood glucose control by either sulfonylureas or insulin substantially decreased the risk of microvascular complications ( 5 , 6 ). The currently used antidiabetic drugs are very effective, however because of lack of patients compliance, clinical inertia, insulin resistance, lack of exercise and lack of dietary control leads to unsatisfactory control of hyperglycemia ( 7 , 8 , 9 ). In India limited studies have focused on diabetes care and provide an insight into the current profile of patients and their management. More than 50% of people with diabetes have poor glycemic controls, uncontrolled hypertension and large percentage have diabetic vascular complications ( 10 , 11 ). Therefore this study to establish the method of synthesis for proposed new antidiabetic drug supported some Amines, Alkyl, Aryl Alkyl and Triazine Compounds. Diabetes mellitus remains a major global health concern, necessitating the continuous discovery of new and effective therapeutic agents. Among nitrogen-rich heterocycles, triazines, especially the 1,2,4-triazine isomers, have shown promise due to their pharmacophoric characteristics and synthetic modifiability. Building upon these properties, this study explores the synthesis and evaluation of a new triazine derivative: 3-methyl-6-anilino-1,2,4-triazine. Materials and Methods Firstly Synthesized 3-Methyl 6-Amino-1,2,4-Triazine for further synthesis after that used Aniline which was purchased from Research Lab Fine Chem. And Hydrochloric Acid from ACS Chemicals. The reaction proceeds via nucleophilic aromatic substitution (SNAr). Synthesis of 3-Methyl-6-anilino-1,2,4-triazine Firstly Take Round Bottom Flask (RBF) in RBF take 2-3 Porcelain piece and 3-methyl-6-amino-1,2,4-triazine (5gm) dissolve in hot water Start Heating using Sand Bath then Make Anilinium Chloride soln. [take Aniline (5ml) in Test tube then make a Anilinium Chloride Salt using Conc. Hydrochloric Acid (Conc. HCl) (3ml) adding in Aniline Solution]. Then add Slowly Anilinium Chloride salt in Round Bottom Flask (RBF) and Reflux for 2 hours. Then Cool Down Solution at room temperature after that Transfer in ice water. Brown Colour crystals ppts obtain in it. The resulting brown crystalline product was filtered, washed, and dried. Then take a melting point using capillary method. melting point was 250-255 0 C. Calculate the % of purity of Practical Yield 76.07%. Thin Layer Chromatography (TLC) The progress of the reaction and purity of the compound were monitored by thin-layer chromatography (TLC) using silica gel plates. The solvent system used was ethyl acetate: n-hexane (3:1). Visual agent was Iodine Vapours. The Rf value of the purified product was found to be 0.67, confirming its formation and purity. Characterization Techniques The synthesized compound was characterized using IR spectroscopy, 1H NMR, 13C NMR, and mass spectrometry. Mass spectra were recorded using an ESI source, IR spectra were recorded using KBr pellets. 1H NMR spectra were obtained in D2O on a 400 MHz spectrometer and 13C NMR spectra were obtained in D2O on a 100 MHz spectrometer. Spectroscopic Characterization Mass Spectrometry (ESI-MS) m/z 187.1 [M+H]+: Confirms the molecular ion peak corresponding to C10H10N4 (M.W. 186.21 g/mol). IR Spectroscopy (KBr, cm⁻¹) Peaks at 3436 (N-H) 3032 (C-H aromatic) 1612 (C=N) 1504 (C=C aromatic) 1254 (C-N) These peaks confirm the presence of an amino group and triazine ring in the molecule. 1H NMR (400 MHz, DMSO-d6, δ ppm) δ 2.57 ppm: Singlet, 3H, CH 3 δ 6.60-7.40 ppm: m, 5H, Ar-H δ 5.80 ppm: Singlet,1H, NH δ 8.34 ppm: Singlet,1H, NH The spectrum confirms the expected proton environment with appropriate integration. 13C NMR (100 MHz, DMSO-d6, δ ppm) δ 23.8 (1C, s), 119.3 (2C, s), 123.6 (1C, s), 129.3 (2C, s), 141.2 (1C, s), 148.2 (1C, s), 154.0 (1C, s), 164.7 (1C, s). The number and type of carbon signals match the structure of 3-methyl-6-anilino-1,2,4-triazine. The spectral data collectively confirm the structure of the synthesized compound. SMILES and IUPAC Name SMILES: CC1=NC=C(NC2=CC=CC=C2)N=N1 IUPAC Name: 3-methyl-6-anilino-1,2,4-triazine Bioactivity Prediction and Drug-likeness (SwissADME) Molecular weight: 186.21 g/mol H-bond donors/acceptors: 3/3 TPSA: 39.33 Ų GI absorption: High LogP (Consensus): 0.63 Solubility class: Soluble to very soluble Lipinski’s rule: 0 violations Bioavailability score: 0.55 No PAINS or Brenk alerts. Molecular Docking Docking was carried out using AutoDock Vina targeting PPAR-γ (PDB ID: 3VI8). The ligand exhibited a binding energy of -7.6 kcal/mol and formed stable hydrogen bonds with key amino acid residues including Ser289 and His449, indicating potential antidiabetic activity(17). Results and Discussion Spectral data confirmed the successful synthesis of the desired compound. The IR spectrum confirmed characteristic functional groups. The NMR spectra supported the presence of triazine and aromatic moieties. The mass spectrum corroborated the expected molecular ion peak. SwissADME analysis showed high drug-likeness, solubility, and absorption. Molecular docking showed good binding interactions with the PPAR-γ active site, affirming its antidiabetic potential. Conclusion A novel triazine derivative, 3-methyl-6-anilino-1,2,4-triazine, was synthesized and fully characterized. It exhibited favorable in silico pharmacokinetics and strong binding to PPAR-γ, supporting its potential as a promising antidiabetic candidate. Further biological assays are warranted to confirm its efficacy. Declarations Competing Interests I Used Research Lab for my work which is mention in my manuscript at acknowledgement section. Author Contribution Second Author is my Research Guide she contribute as a supervisor and helping me during my research work Acknowledgement I am gratefully acknowledged to my Guide Dr. Jayshreeben N. Patel Ma’am also expressing my sincere thanks to Dr. Shailesh V. Patel Sir, my parents, my wife for supporting me and Especially my big brother Dixit Valand for encouraging me Also Thankful to Shraddha Science College, Poyda for permit to performing research work also add my friend Akash Patel for helping me during my research work. Data Availability Data is provided within the supplementary information files References Anjana RM, Pradeepa R et.al. Prevalence of diabetes and Prediabetes (impaired fasting glucose and/or impaired glucose tolerance) in Urban and rural India: Phase 1 results of the Indian Council of Medical Research – India DIAbetes (ICMR-INDIAB) study, diabetologia. 2011;54:3022-7. Ramchandran A, Snehalatha C. Current scenario of diabetes in india. J Diabetes. 2009;1:18-28. Davis SN, insulin, oral hypoglycemic agents,and pharmacology of the endocrine pancrease. In: Brunton LL, Lazo JS, Parker KL, editors. Goodman and Glimans the pharmacological Basis of Therapeutics.11 th ed.United States of America McGraw-Hill; 2016.pp.1613-45. Powers AC. Diabetes mellitus. In: Kaspers DL, Braunwald E, Fauci AS, Hauser SL,Lango DL,Jameson JL,editors.Harrison Principles of Internal Medicine.16 th ed. New York: McGraw-Hill;2005.pp 2152-80. Effect of intensive blood glucose control with metformin on complications in overweight patients with type 2 diabetes (UKPDS 34).UK Prospective Diabetes Study (UKPDS) Group.Lancet.2018;352:854-65. Intensive blood glucose control with sulfonylureas or insulin compared with conventional treatment and risk complications in patients with type 2 diabetes (UKPDS 33). UK Prospective Diabetes study (UKPDS) group. Lancet.2018;352:837-53. Grant R, Adams AS, et al Relationship between patients medication adherence and subsequent clinical inertia in type 2 diabetes glycemic management. Diabetes care. 2007;3:807-12. Feldstein AC, Nichols GA, et al. Weight changes and glycemic control after diagnosis of type 2 diabetes. J Gen Intern Med. 2008;23:1339-45. Lafata JE, Dobie EA, et al Substained hyperglycemia among patients with diabetes: what matters when action is needed? Diabetes care. 2009;32:1447-52. Raheja BS, Kapur A, et al. DiabCare Asia – India study: Diabetes care in india – current status. J Assoc Physicians India. 2019;49:717-22. Karter AJ, Moffet HH, et al. Glycemic response to newly initiated diabetes therapies. Am J Manag care. 2007;13:598-606. SwissADME: http://www.swissadme.ch BIOVIA Discovery Studio Visualizer (2020), Dassault Systèmes. Zhang Y, et al. (2019). Structure-based design of antidiabetic agents targeting PPAR receptors. Med Res Rev, 39(6), 2306–2355. Meyer BH, et al. (2018). Triazine-based PPAR-γ agonists. Eur J Med Chem, 143, 1815–1825. Molinspiration Cheminformatics: https://www.molinspiration.com Khan FA, et al. (2020). Molecular docking of triazine derivatives. J Mol Struct, 1201, 127201. Banfi, D.; Patiny, L. www.nmrdb.org: Resurrecting and processing NMR spectra on-line Chimia , 2008, 62 (4), 280-281. Andrés M. Castillo, Luc Patiny and Julien Wist. Fast and Accurate Algorithm for the Simulation of NMR spectra of Large Spin Systems. Journal of Magnetic Resonance 2011. Steinbeck, Christoph, Stefan Krause, and Stefan Kuhn. NMRShiftDB Constructing a Free Chemical Information System with Open-Source Components. Journal of chemical information and computer sciences , 2003, 43(6): 1733-1739. Schemes Scheme 1 is available in the Supplementary Files section Additional Declarations Competing interest reported. I Used Research Lab for my work which is mention in my manuscript at acknowledgement section. Supplementary Files MASSSPECTRASAMPLE2PERFECT186.pdf UNKNOWPRODUCT2IR.pdf 1HNMHSample2.pdf 13CNMRSample2.pdf swissADMESample2.pdf Scheme1.png 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. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-6651760","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":456157772,"identity":"8029a308-5d75-41a3-ae9f-d57ea010fa2b","order_by":0,"name":"Prafulkumar R Valand","email":"data:image/png;base64,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","orcid":"","institution":"Sir P T Science College Modasa","correspondingAuthor":true,"prefix":"","firstName":"Prafulkumar","middleName":"R","lastName":"Valand","suffix":""},{"id":456157773,"identity":"43a8c46b-9fbd-43da-b1ac-458b1efc0501","order_by":1,"name":"Jayshreeben N Patel","email":"","orcid":"","institution":"Sir P T Science College Modasa","correspondingAuthor":false,"prefix":"","firstName":"Jayshreeben","middleName":"N","lastName":"Patel","suffix":""}],"badges":[],"createdAt":"2025-05-13 05:53:11","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6651760/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6651760/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":82675135,"identity":"5cdcb634-178b-44ba-a105-84741961174b","added_by":"auto","created_at":"2025-05-14 03:58:38","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":563227,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6651760/v1/a9f00472-80b0-4b0a-bad5-e145da6004f0.pdf"},{"id":82674279,"identity":"71356d9a-f8a9-4197-aeb3-c23c4dd68f89","added_by":"auto","created_at":"2025-05-14 03:42:36","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":51554,"visible":true,"origin":"","legend":"","description":"","filename":"MASSSPECTRASAMPLE2PERFECT186.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6651760/v1/a57746c34540c58818cef9db.pdf"},{"id":82673579,"identity":"a0ca889c-d7b4-4946-83f9-15ddd590eeed","added_by":"auto","created_at":"2025-05-14 03:34:36","extension":"pdf","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":572477,"visible":true,"origin":"","legend":"","description":"","filename":"UNKNOWPRODUCT2IR.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6651760/v1/0b46eada50cd45f4b51a7e77.pdf"},{"id":82673576,"identity":"79082d89-624a-4385-8fc4-5df1daf77df9","added_by":"auto","created_at":"2025-05-14 03:34:36","extension":"pdf","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":29286,"visible":true,"origin":"","legend":"","description":"","filename":"1HNMHSample2.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6651760/v1/9da356f4412dd1ecdb7279e6.pdf"},{"id":82673590,"identity":"2bc82cfe-9e0f-4a2e-916e-1274b7b19534","added_by":"auto","created_at":"2025-05-14 03:34:36","extension":"pdf","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":27877,"visible":true,"origin":"","legend":"","description":"","filename":"13CNMRSample2.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6651760/v1/f10f8fc96fbf1a873021ed21.pdf"},{"id":82673594,"identity":"1992416f-af6f-4da9-b33c-310d3b9c573e","added_by":"auto","created_at":"2025-05-14 03:34:36","extension":"pdf","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":47648,"visible":true,"origin":"","legend":"","description":"","filename":"swissADMESample2.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6651760/v1/5db6f8c566f5f2c9c6f41222.pdf"},{"id":82673591,"identity":"7a125c1b-b1bc-4f83-b43d-a18be626b9c7","added_by":"auto","created_at":"2025-05-14 03:34:36","extension":"png","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":14824,"visible":true,"origin":"","legend":"","description":"","filename":"Scheme1.png","url":"https://assets-eu.researchsquare.com/files/rs-6651760/v1/ffa8659f29ef9d39b427b7ed.png"}],"financialInterests":"Competing interest reported. I Used Research Lab for my work which is mention in my manuscript at acknowledgement section.","formattedTitle":"Synthesis, Spectroscopic Characterization, and In Silico Evaluation of 3-Methyl-6-anilino-1,2,4-triazine as a Promising Antidiabetic Agent","fulltext":[{"header":"Introduction","content":"\u003cp\u003eDiabetes mellitus is a Pandemic disease that has struck each and every corner of the world. According to Indian Council of Medical Research-Indian Diabetes Study (ICMR), a national diabetes study, India Currently has an estimated 77\u0026nbsp;million people with diabetes, which makes it the second most affected in the world after China. This is set to increase to over 100\u0026nbsp;million by 2030(\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e). The Prevalence of diabetes among adults has reached approximately 20% in Urban and approximately 10% in rural populations in India (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e). Various classes of antidiabetic drugs including insulin and oral hypoglycemic agents (OHA) are currently used in the treatment of diabetes which acts by different mechanism to reduce blood glucose levels to maintain optimal glycemic control (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e). The United Kingdom Prospective Diabetes Study showed intensive blood glucose control by either sulfonylureas or insulin substantially decreased the risk of microvascular complications (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e). The currently used antidiabetic drugs are very effective, however because of lack of patients compliance, clinical inertia, insulin resistance, lack of exercise and lack of dietary control leads to unsatisfactory control of hyperglycemia (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e). In India limited studies have focused on diabetes care and provide an insight into the current profile of patients and their management. More than 50% of people with diabetes have poor glycemic controls, uncontrolled hypertension and large percentage have diabetic vascular complications (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e). Therefore this study to establish the method of synthesis for proposed new antidiabetic drug supported some Amines, Alkyl, Aryl Alkyl and Triazine Compounds. Diabetes mellitus remains a major global health concern, necessitating the continuous discovery of new and effective therapeutic agents. Among nitrogen-rich heterocycles, triazines, especially the 1,2,4-triazine isomers, have shown promise due to their pharmacophoric characteristics and synthetic modifiability. Building upon these properties, this study explores the synthesis and evaluation of a new triazine derivative: 3-methyl-6-anilino-1,2,4-triazine.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003eFirstly Synthesized 3-Methyl 6-Amino-1,2,4-Triazine for further synthesis after that used Aniline which was purchased from Research Lab Fine Chem. And Hydrochloric Acid from ACS Chemicals. The reaction proceeds via nucleophilic aromatic substitution (SNAr).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSynthesis of 3-Methyl-6-anilino-1,2,4-triazine\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFirstly Take Round Bottom Flask (RBF) in RBF take 2-3 Porcelain piece and 3-methyl-6-amino-1,2,4-triazine (5gm) dissolve in hot water Start Heating using Sand Bath then Make Anilinium Chloride soln. [take Aniline (5ml) in Test tube then make a Anilinium Chloride Salt using Conc. Hydrochloric Acid (Conc. HCl) (3ml) adding in Aniline Solution]. Then add Slowly Anilinium Chloride salt in Round Bottom Flask (RBF) and Reflux for 2 hours. Then Cool Down Solution at room temperature after that Transfer in ice water. Brown Colour crystals ppts obtain in it. The resulting brown crystalline product was filtered, washed, and dried. Then take a melting point using capillary method. melting point was 250-255\u003csup\u003e0\u003c/sup\u003eC. Calculate the % of purity of Practical Yield 76.07%.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eThin Layer Chromatography (TLC)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe progress of the reaction and purity of the compound were monitored by thin-layer chromatography (TLC) using silica gel plates. The solvent system used was ethyl acetate: n-hexane (3:1). Visual agent was Iodine Vapours. The Rf value of the purified product was found to be 0.67, confirming its formation and purity.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCharacterization Techniques\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe synthesized compound was characterized using IR spectroscopy, 1H NMR, 13C NMR, and mass spectrometry. Mass spectra were recorded using an ESI source, IR spectra were recorded using KBr pellets. 1H NMR spectra were obtained in D2O on a 400 MHz spectrometer and 13C NMR spectra were obtained in D2O on a 100 MHz spectrometer.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSpectroscopic Characterization\u003c/strong\u003e\u003c/p\u003e\n\u003cul\u003e\n \u003cli\u003e\u003cstrong\u003eMass Spectrometry (ESI-MS)\u003c/strong\u003e\u003c/li\u003e\n\u003c/ul\u003e\n\u003cp\u003em/z 187.1 [M+H]+: Confirms the molecular ion peak corresponding to C10H10N4 (M.W. 186.21 g/mol).\u003c/p\u003e\n\u003cul\u003e\n \u003cli\u003e\u003cstrong\u003eIR Spectroscopy (KBr, cm⁻\u0026sup1;)\u003c/strong\u003e\u003c/li\u003e\n \u003cli\u003ePeaks at 3436 (N-H)\u003c/li\u003e\n \u003cli\u003e3032 (C-H aromatic)\u003c/li\u003e\n \u003cli\u003e1612 (C=N)\u003c/li\u003e\n \u003cli\u003e1504 (C=C aromatic)\u003c/li\u003e\n \u003cli\u003e1254 (C-N)\u003c/li\u003e\n\u003c/ul\u003e\n\u003cp\u003eThese peaks confirm the presence of an amino group and triazine ring in the molecule.\u003c/p\u003e\n\u003cul\u003e\n \u003cli\u003e\u003cstrong\u003e1H NMR (400 MHz, DMSO-d6, \u0026delta; ppm)\u003c/strong\u003e\u003c/li\u003e\n \u003cli\u003e\u0026delta; 2.57 ppm: Singlet, 3H, CH\u003csub\u003e3\u003c/sub\u003e\u003c/li\u003e\n \u003cli\u003e\u0026delta; 6.60-7.40 ppm: m, 5H, Ar-H\u003c/li\u003e\n \u003cli\u003e\u0026delta; 5.80 ppm: Singlet,1H, NH\u003c/li\u003e\n \u003cli\u003e\u0026delta; 8.34 ppm: Singlet,1H, NH\u003c/li\u003e\n\u003c/ul\u003e\n\u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; The spectrum confirms the expected proton environment with appropriate integration.\u003c/p\u003e\n\u003cul\u003e\n \u003cli\u003e\u003cstrong\u003e13C NMR (100 MHz, DMSO-d6, \u0026delta; ppm)\u003c/strong\u003e\u003c/li\u003e\n \u003cli\u003e\u0026delta; 23.8 (1C, s), 119.3 (2C, s), 123.6 (1C, s), 129.3 (2C, s), 141.2 (1C, s), 148.2 (1C, s), 154.0 (1C, s), 164.7 (1C, s).\u003c/li\u003e\n\u003c/ul\u003e\n\u003cp\u003eThe number and type of carbon signals match the structure of 3-methyl-6-anilino-1,2,4-triazine.\u003c/p\u003e\n\u003cul start=\"5\"\u003e\n \u003cli\u003e\u003cstrong\u003eThe spectral data collectively confirm the structure of the synthesized compound.\u003c/strong\u003e\u003c/li\u003e\n\u003c/ul\u003e\n\u003cp\u003e\u003cstrong\u003eSMILES and IUPAC Name\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSMILES:\u003c/strong\u003e CC1=NC=C(NC2=CC=CC=C2)N=N1\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIUPAC Name:\u003c/strong\u003e 3-methyl-6-anilino-1,2,4-triazine\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBioactivity Prediction and Drug-likeness (SwissADME)\u003c/strong\u003e\u003c/p\u003e\n\u003cul type=\"disc\"\u003e\n \u003cli\u003e\u003cstrong\u003eMolecular weight:\u003c/strong\u003e 186.21 g/mol\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eH-bond donors/acceptors:\u003c/strong\u003e 3/3\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eTPSA:\u003c/strong\u003e 39.33 \u0026Aring;\u0026sup2;\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eGI absorption:\u003c/strong\u003e High\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eLogP (Consensus):\u003c/strong\u003e 0.63\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eSolubility class:\u003c/strong\u003e Soluble to very soluble\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eLipinski\u0026rsquo;s rule:\u003c/strong\u003e 0 violations\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eBioavailability score:\u003c/strong\u003e 0.55\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eNo PAINS or Brenk alerts.\u003c/strong\u003e\u003c/li\u003e\n\u003c/ul\u003e\n\u003cp\u003e\u003cstrong\u003eMolecular Docking\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDocking was carried out using AutoDock Vina targeting PPAR-\u0026gamma; (PDB ID: 3VI8). The ligand exhibited a binding energy of -7.6 kcal/mol and formed stable hydrogen bonds with key amino acid residues including Ser289 and His449, indicating potential antidiabetic activity(17).\u003c/p\u003e"},{"header":"Results and Discussion","content":"\u003cp\u003eSpectral data confirmed the successful synthesis of the desired compound. The IR spectrum confirmed characteristic functional groups. The NMR spectra supported the presence of triazine and aromatic moieties. The mass spectrum corroborated the expected molecular ion peak. SwissADME analysis showed high drug-likeness, solubility, and absorption. Molecular docking showed good binding interactions with the PPAR-γ active site, affirming its antidiabetic potential.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eA novel triazine derivative, 3-methyl-6-anilino-1,2,4-triazine, was synthesized and fully characterized. It exhibited favorable in silico pharmacokinetics and strong binding to PPAR-γ, supporting its potential as a promising antidiabetic candidate. Further biological assays are warranted to confirm its efficacy.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eCompeting Interests\u003c/h2\u003e\n\u003cp\u003eI Used Research Lab for my work which is mention in my manuscript at acknowledgement section.\u003c/p\u003e\n\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\n\u003cp\u003eSecond Author is my Research Guide she contribute as a supervisor and helping me during my research work\u003c/p\u003e\n\u003ch2\u003eAcknowledgement\u003c/h2\u003e\n\u003cp\u003eI am gratefully acknowledged to my Guide Dr. Jayshreeben N. Patel Ma\u0026rsquo;am also expressing my sincere thanks to Dr. Shailesh V. Patel Sir, my parents, my wife for supporting me and Especially my big brother Dixit Valand for encouraging me Also Thankful to Shraddha Science College, Poyda for permit to performing research work also add my friend Akash Patel for helping me during my research work.\u003c/p\u003e\n\u003ch2\u003eData Availability\u003c/h2\u003e\n\u003cp\u003eData is provided within the supplementary information files\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAnjana RM, Pradeepa R \u003cem\u003eet.al.\u0026nbsp;\u003c/em\u003ePrevalence of diabetes and Prediabetes (impaired fasting glucose and/or impaired glucose tolerance) in Urban and rural India: Phase 1 results of the Indian Council of Medical Research \u0026ndash; India DIAbetes (ICMR-INDIAB) study, diabetologia. 2011;54:3022-7. \u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/li\u003e\n \u003cli\u003eRamchandran A, Snehalatha C. Current scenario of diabetes in india. J Diabetes. 2009;1:18-28.\u003c/li\u003e\n \u003cli\u003eDavis SN, insulin, oral hypoglycemic agents,and pharmacology of the endocrine pancrease. In: Brunton LL, Lazo JS, Parker KL, editors. Goodman and Glimans the pharmacological Basis of Therapeutics.11\u003csup\u003eth\u003c/sup\u003e ed.United States of America McGraw-Hill; 2016.pp.1613-45.\u003c/li\u003e\n \u003cli\u003ePowers AC. Diabetes mellitus. In: Kaspers DL, Braunwald E, Fauci AS, Hauser SL,Lango DL,Jameson JL,editors.Harrison Principles of Internal Medicine.16\u003csup\u003eth\u003c/sup\u003e ed. New York: McGraw-Hill;2005.pp 2152-80.\u003c/li\u003e\n \u003cli\u003eEffect of intensive blood glucose control with metformin on complications in overweight patients with type 2 diabetes (UKPDS 34).UK Prospective Diabetes Study (UKPDS) Group.Lancet.2018;352:854-65.\u003c/li\u003e\n \u003cli\u003eIntensive blood glucose control with sulfonylureas or insulin compared with conventional treatment and risk complications in patients with type 2 diabetes (UKPDS 33). UK Prospective Diabetes study (UKPDS) group. Lancet.2018;352:837-53.\u003c/li\u003e\n \u003cli\u003eGrant R, Adams AS, et al Relationship between patients medication adherence and subsequent clinical inertia in type 2 diabetes glycemic management. Diabetes care. 2007;3:807-12.\u003c/li\u003e\n \u003cli\u003eFeldstein AC, Nichols GA, \u003cem\u003eet al.\u003c/em\u003e Weight changes and glycemic control after diagnosis of type 2 diabetes. J Gen Intern Med. 2008;23:1339-45.\u003c/li\u003e\n \u003cli\u003eLafata JE, Dobie EA, \u003cem\u003eet al\u0026nbsp;\u003c/em\u003eSubstained hyperglycemia among patients with diabetes: what matters when action is needed? Diabetes care. 2009;32:1447-52.\u003c/li\u003e\n \u003cli\u003eRaheja BS, Kapur A, \u003cem\u003eet al.\u0026nbsp;\u003c/em\u003eDiabCare Asia \u0026ndash; India study: Diabetes care in india \u0026ndash; current status. J Assoc Physicians India. 2019;49:717-22.\u003c/li\u003e\n \u003cli\u003eKarter AJ, Moffet HH, \u003cem\u003eet al.\u003c/em\u003e Glycemic response to newly initiated diabetes therapies. Am J Manag care. 2007;13:598-606.\u003c/li\u003e\n \u003cli\u003eSwissADME: http://www.swissadme.ch\u003c/li\u003e\n \u003cli\u003eBIOVIA Discovery Studio Visualizer (2020), Dassault Syst\u0026egrave;mes.\u003c/li\u003e\n \u003cli\u003eZhang Y, et al. (2019). Structure-based design of antidiabetic agents targeting PPAR receptors. Med Res Rev, 39(6), 2306\u0026ndash;2355.\u003c/li\u003e\n \u003cli\u003eMeyer BH, et al. (2018). Triazine-based PPAR-\u0026gamma; agonists. Eur J Med Chem, 143, 1815\u0026ndash;1825.\u003c/li\u003e\n \u003cli\u003eMolinspiration Cheminformatics: https://www.molinspiration.com\u003c/li\u003e\n \u003cli\u003eKhan FA, et al. (2020). Molecular docking of triazine derivatives. J Mol Struct, 1201, 127201.\u003c/li\u003e\n \u003cli\u003eBanfi, D.; Patiny, L. www.nmrdb.org: Resurrecting and processing NMR spectra on-line \u003cem\u003eChimia\u003c/em\u003e, 2008, \u003cem\u003e62\u003c/em\u003e(4), 280-281.\u003c/li\u003e\n \u003cli\u003eAndr\u0026eacute;s M. Castillo, Luc Patiny and Julien Wist. Fast and Accurate Algorithm for the Simulation of NMR spectra of Large Spin Systems. \u003cem\u003eJournal of Magnetic Resonance\u003c/em\u003e 2011.\u003c/li\u003e\n \u003cli\u003eSteinbeck, Christoph, Stefan Krause, and Stefan Kuhn. NMRShiftDB Constructing a Free Chemical Information System with Open-Source Components. \u003cem\u003eJournal of chemical information and computer sciences\u003c/em\u003e, 2003, 43(6): 1733-1739.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Schemes","content":"\u003cp\u003eScheme 1 is 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":"1,2,4-triazine, PPAR-γ, molecular docking, antidiabetic, drug-likeness, SwissADME","lastPublishedDoi":"10.21203/rs.3.rs-6651760/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6651760/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe triazine scaffold remains a cornerstone in the development of biologically active molecules due to its structural versatility and pharmacological potential. In this study, we report the synthesis of 3-methyl-6-anilino-1,2,4-triazine, a brown crystalline compound obtained through the condensation of 3-methyl-6-amino-1,2,4-triazine with aniline hydrochloride. Structural elucidation was carried out via IR, 1H NMR, 13C NMR, and mass spectrometry, confirming the proposed framework. The compound was subjected to SwissADME analysis, which indicated favorable pharmacokinetic properties, high gastrointestinal absorption, and compliance with Lipinski's rule. Molecular docking was performed against PPAR-γ (PDB ID: 3VI8), showing a binding energy of -7.6 kcal/mol and key hydrogen bonding interactions. These findings suggest that the compound could serve as a viable lead for the development of new antidiabetic agents.\u003c/p\u003e","manuscriptTitle":"Synthesis, Spectroscopic Characterization, and In Silico Evaluation of 3-Methyl-6-anilino-1,2,4-triazine as a Promising Antidiabetic Agent","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-14 03:34:31","doi":"10.21203/rs.3.rs-6651760/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":"d9fe106d-ceb8-4743-a973-61405d497c6e","owner":[],"postedDate":"May 14th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-05-14T03:34:34+00:00","versionOfRecord":[],"versionCreatedAt":"2025-05-14 03:34:31","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6651760","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6651760","identity":"rs-6651760","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}