{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T01:55:18Z","timestamp":1760234118811,"version":"build-2065373602"},"reference-count":49,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2021,3,25]],"date-time":"2021-03-25T00:00:00Z","timestamp":1616630400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004281","name":"Narodowe Centrum Nauki","doi-asserted-by":"publisher","award":["Preludium (Grant No. 2018\/31\/N\/NZ7\/02867)"],"award-info":[{"award-number":["Preludium (Grant No. 2018\/31\/N\/NZ7\/02867)"]}],"id":[{"id":"10.13039\/501100004281","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>About 70 million people suffer from epilepsy\u2014a chronic neurodegenerative disease. In most cases, the cause of the disease is unknown, but epilepsy can also develop as the result of a stroke, trauma to the brain, or the use of psychotropic substances. The treatment of epilepsy is mainly based on the administration of anticonvulsants, which the patient must most often use throughout their life. Despite significant progress in research on antiepileptic drugs, about 30% of patients still have drug-resistant epilepsy, which is insensitive to pharmacotherapy used so far. In our recent studies, we have shown that 4-alkyl-5-aryl-1,2,4-triazole-3-thiones act on the voltage-gated sodium channels and exhibit anticonvulsant activity in an MES (maximal electroshock-induced seizure) and 6Hz test in mice. Previous studies have shown their beneficial toxic and pharmacological profile, but their effect on a living organism during chronic use is still unknown. In the presented study, on the basis of the previously conducted tests and the PAMPA (parallel artificial membrane permeability assay) BBB (blood\u2013brain barrier) test, we selected one 1,2,4-triazole-3-thione derivative\u2014TP-315\u2014for further studies aimed at assessing the impact of its chronic use on a living organism. After long-term administration of TP-315 to Albino Swiss mice, its effect on the functional parameters of internal organs was assessed by performing biochemical, morphological, and histopathological examinations. It was also determined whether the tested compound inhibits selected isoforms of the CYP450 enzyme system. On the basis of the conducted tests, it was found that TP-315 does not show nephrotoxic nor hepatotoxic effects and does not cause changes in hematological parameters. In vitro tests showed that TP-315 did not inhibit CYP2B6, CYP2D6, CYP3A4, or CYP3A5 enzymes at the concentration found in the serum of mice subjected to long-term exposure to this compound.<\/jats:p>","DOI":"10.3390\/ijms22073358","type":"journal-article","created":{"date-parts":[[2021,3,25]],"date-time":"2021-03-25T14:51:12Z","timestamp":1616683872000},"page":"3358","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Effect of Chronic Administration of 5-(3-chlorophenyl)-4-Hexyl-2,4 -Dihydro-3H-1,2,4-Triazole-3-Thione (TP-315)\u2014A New Anticonvulsant Drug Candidate\u2014On Living Organisms"],"prefix":"10.3390","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8082-535X","authenticated-orcid":false,"given":"Anna","family":"Makuch-Kocka","sequence":"first","affiliation":[{"name":"Department of Pharmacology, Faculty of Health Sciences, Medical University of Lublin, 20-093 Lublin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marta","family":"Andres-Mach","sequence":"additional","affiliation":[{"name":"Isobolographic Analysis Laboratory, Institute of Rural Health, 20-090 Lublin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Miros\u0142aw","family":"Zagaja","sequence":"additional","affiliation":[{"name":"Isobolographic Analysis Laboratory, Institute of Rural Health, 20-090 Lublin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5816-4521","authenticated-orcid":false,"given":"Anna","family":"\u015amiech","sequence":"additional","affiliation":[{"name":"Sub-Department of Pathomorphology and Forensic Veterinary Medicine, Department and Clinic of Animal Internal Diseases, University of Life Sciences in Lublin, 20-612 Lublin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6660-1384","authenticated-orcid":false,"given":"Magdalena","family":"Pizo\u0144","sequence":"additional","affiliation":[{"name":"Department of Analytical Chemistry, Faculty of Pharmacy, Medical University of Lublin, 20-093 Lublin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jolanta","family":"Flieger","sequence":"additional","affiliation":[{"name":"Department of Analytical Chemistry, Faculty of Pharmacy, Medical University of Lublin, 20-093 Lublin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0891-5419","authenticated-orcid":false,"given":"Judyta","family":"Cielecka-Piontek","sequence":"additional","affiliation":[{"name":"Department of Pharmacognosy, Faculty of Pharmacy, Poznan University of Medical Sciences, 61-781 Pozna\u0144, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8162-8435","authenticated-orcid":false,"given":"Tomasz","family":"Plech","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Faculty of Health Sciences, Medical University of Lublin, 20-093 Lublin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,3,25]]},"reference":[{"key":"ref_1","first-page":"535","article-title":"Prevalence, type of epilepsy and use of antiepileptic drugs in primary care","volume":"60","year":"2015","journal-title":"Rev. Neurol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1097\/ACO.0000000000000376","article-title":"Recent advances in epilepsy management","volume":"29","author":"Ernst","year":"2016","journal-title":"Curr. Opin. Anaesthesiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.neurol.2015.11.003","article-title":"Epidemiology of epilepsy","volume":"172","author":"Behr","year":"2016","journal-title":"Rev. Neurol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"72","DOI":"10.21784\/IwP.2016.018","article-title":"Epilepsy-the Social Disease of the 21st Century","volume":"3","author":"Filipska","year":"2016","journal-title":"Innow. Piel\u0119gniarstwie Naukach Zdrowiu"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"314","DOI":"10.1056\/NEJM200002033420503","article-title":"Early identification of refractory epilepsy","volume":"342","author":"Kwan","year":"2000","journal-title":"N. Engl. J. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1159\/000222775","article-title":"Refractory epilepsy: A clinically oriented review","volume":"62","author":"Beleza","year":"2009","journal-title":"Eur. Neurol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1684\/epd.2020.1201","article-title":"How to understand and address the cultural aspects and consequences of diagnosis of epilepsy, including stigma","volume":"22","author":"Braga","year":"2020","journal-title":"Epileptic Disord."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Kapro\u0144, B., \u0141uszczki, J.J., Siwek, A., Karcz, T., Nowak, G., Zagaja, M., Andres-Mach, M., Stasi\u0142owicz, A., Cielecka-Piontek, J., and Kocki, J. (2020). Preclinical evaluation of 1,2,4-triazole-based compounds targeting voltage-gated sodium channels (VGSCs) as promising anticonvulsant drug candidates. Bioorg. Chem., 94.","DOI":"10.1016\/j.bioorg.2019.103355"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1007\/s43440-019-00044-7","article-title":"Anticonvulsant and neurotoxic effects of a novel 1,2,4-triazole-3-thione derivative (TPF-34) and its isobolographic interaction profile with classical antiepileptic drugs in mice","volume":"72","author":"Lepiech","year":"2020","journal-title":"Pharm. Rep."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1016\/j.ejmech.2012.11.026","article-title":"Synthesis, characterization and preliminary anticonvulsant evaluation of some 4-alkyl-1,2,4-triazoles","volume":"60","author":"Plech","year":"2013","journal-title":"Eur. J. Med. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"690","DOI":"10.1016\/j.ejmech.2014.09.034","article-title":"Studies on the anticonvulsant activity of 4-alkyl-1,2,4-triazole-3-thiones and their effect on GABAergic system","volume":"86","author":"Plech","year":"2014","journal-title":"Eur. J. Med. Chem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"993","DOI":"10.1080\/14756366.2020.1748026","article-title":"1,2,4-Triazole-based anticonvulsant agents with additional ROS scavenging activity are effective in a model of pharmacoresistant epilepsy","volume":"35","author":"Czarnomysy","year":"2020","journal-title":"J. Enzym. Inhib. Med. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"11279","DOI":"10.3390\/molecules190811279","article-title":"Studies on the anticonvulsant activity and influence on GABA-ergic neurotransmission of 1,2,4-triazole-3-thione- based compounds","volume":"19","author":"Plech","year":"2014","journal-title":"Molecules"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/j.ejphar.2012.06.023","article-title":"Effect of 4-(4-bromophenyl)-5-(3-chlorophenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione on the anticonvulsant action of different classical antiepileptic drugs in the mouse maximal electroshock-induced seizure model","volume":"690","author":"Plech","year":"2012","journal-title":"Eur. J. Pharm."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"970","DOI":"10.1016\/S1734-1140(12)70893-9","article-title":"Influence of 5-(3-chlorophenyl)-4-(4-methylphenyl)-2,-dihydro-3H-1,2,4-triazole-3-thione on the anticonvulsant action of four classical antiepileptic drugs in the mouse maximal electroshock-induced seizure model","volume":"64","author":"Plech","year":"2012","journal-title":"Pharm. Rep."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1016\/j.ejmech.2016.01.009","article-title":"The importance of triazole scaffold in the development of anticonvulsant agents","volume":"109","author":"Ayati","year":"2016","journal-title":"Eur. J. Med. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1080\/14756366.2017.1423068","article-title":"Recent developments on triazole nucleus in anticonvulsant compounds: A review","volume":"33","author":"Song","year":"2018","journal-title":"J. Enzym. Inhib. Med. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.ejps.2018.12.018","article-title":"Development of the 1,2,4-triazole-based anticonvulsant drug candidates acting on the voltage-gated sodium channels. Insights from in-vivo, in-vitro, and in-silico studies","volume":"129","author":"Siwek","year":"2019","journal-title":"Eur. J. Pharm. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"973","DOI":"10.1208\/s12248-017-0096-2","article-title":"Breaking bad: The structure and function of the blood-brain barrier in epilepsy","volume":"19","author":"Han","year":"2017","journal-title":"AAPS J."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"4943","DOI":"10.1016\/j.biomaterials.2011.03.031","article-title":"Targeting the brain with PEG-PLGA nanoparticles modified with phage-displayed peptides","volume":"32","author":"Li","year":"2011","journal-title":"Biomaterials"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Galdeano, C., Coquelle, N., Cieslikiewicz-Bouet, M., Bartolini, M., P\u00e9rez, B., Clos, M., Silman, I., Jean, L., Colletier, J.P., and Renard, P.Y. (2018). Increasing Polarity in Tacrine and Huprine Derivatives: Potent Anticholinesterase Agents for the Treatment of Myasthenia Gravis. Molecules, 23.","DOI":"10.3390\/molecules23030634"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"741","DOI":"10.7150\/ijms.20001","article-title":"Molecular mechanism of action and safety of 5-(3-chlorophenyl)-4-hexyl-2,4-dihydro-3H-1,2,4-triazole-3-thione\u2014A novel anticonvulsant drug candidate","volume":"14","author":"Paneth","year":"2017","journal-title":"Int. J. Med. Sci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1239","DOI":"10.1111\/j.1528-1167.2008.01561.x","article-title":"Antiepileptic drugs-best practice guidelines for therapeutic drug monitoring: A position paper by the subcommission on therapeutic drug monitoring, ILAE Commission on Therapeutic Strategies","volume":"49","author":"Patsalos","year":"2008","journal-title":"Epilepsia"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"971","DOI":"10.1248\/bpb.35.971","article-title":"Correlation between plasma ammonia level and serum trough concentration of free valproic acid in patients with epilepsy","volume":"35","author":"Itoh","year":"2012","journal-title":"Biol. Pharm. Bull."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.dld.2005.06.004","article-title":"Suspected drug-induced liver fatalities reported to the WHO database","volume":"38","author":"Bjornsson","year":"2006","journal-title":"Dig. Liver Dis."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1517\/14656566.9.2.285","article-title":"Oral valproic acid for epilepsy-long-term experience in therapy and side effects","volume":"9","author":"Gerstner","year":"2008","journal-title":"Expert Opin. Pharmacother."},{"key":"ref_27","first-page":"239","article-title":"Effects of Carbamazepine and Valproate on Serum Aspartate Aminotransferase, Alanine Aminotransferase and Gamma","volume":"17","author":"Hasanbegovic","year":"2017","journal-title":"Glutamyltransferase Child. Med. Arch."},{"key":"ref_28","first-page":"705","article-title":"Carbamazepine adverse drug reactions","volume":"11","author":"LLerena","year":"2008","journal-title":"Expert Rev. Clin. Pharmacol."},{"key":"ref_29","first-page":"233","article-title":"Drug-induced hepatotoxicity: A review","volume":"2","author":"Pandit","year":"2012","journal-title":"J. Appl. Pharm. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"642","DOI":"10.1111\/j.1365-2230.2006.02185.x","article-title":"Epidemiological study of severe cutaneous adverse drug reactions in a city district of China","volume":"31","author":"Li","year":"2006","journal-title":"Clin. Exp. Dermatol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/S0920-1211(02)00182-1","article-title":"Overtreatment in epilepsy: Adverse consequences and mechanisms","volume":"52","author":"Perucca","year":"2002","journal-title":"Epilepsy Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1385\/NCC:3:2:161","article-title":"Phenytoin poisoning","volume":"3","author":"Craig","year":"2005","journal-title":"Neurocrit Care"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"792","DOI":"10.1016\/S1474-4422(12)70153-9","article-title":"Adverse effects of antiepileptic drugs","volume":"11","author":"Perucca","year":"2012","journal-title":"Lancet Neurol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1046\/j.1365-2710.1999.00201.x","article-title":"Clinically significant pharmacokinetic drug interactions between antiepileptic drugs","volume":"24","author":"Tanaka","year":"1999","journal-title":"J. Clin. Pharm. Ther."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1111\/j.1365-2125.2005.02529.x","article-title":"Clinically relevant drug interactions with antiepileptic drug","volume":"61","author":"Perucca","year":"2005","journal-title":"Br. J. Clin. Pharmacol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1720","DOI":"10.2146\/ajhp070508","article-title":"Potential drug-drug interactions with antiepileptic drugs in medical recepients","volume":"65","author":"Dickson","year":"2008","journal-title":"Am. J. Health Syst. Pharm."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1046\/j.1528-1157.2002.13001.x","article-title":"The importance of drug interactions in epilepsy therapy","volume":"43","author":"Patsalos","year":"2002","journal-title":"Epilepsia"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"722","DOI":"10.1097\/01.ftd.0000179854.16846.67","article-title":"The pharmacokinetics and interactions of new antiepileptic drugs","volume":"27","author":"Bialer","year":"2005","journal-title":"Drug Monit."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1007\/BF03040747","article-title":"Arzneimittelwechselwirkungen mit antiepileptica","volume":"116","author":"Turnheim","year":"2004","journal-title":"Wien Klin. Wochenschr."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1021\/jf048413x","article-title":"Determination of pesticides in apple-based infant foods using liquid chromatography electrospray ionization tandem mass spectrometry","volume":"53","author":"Wang","year":"2005","journal-title":"J. Agric. Food Chem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/S0021-9673(01)01429-7","article-title":"Matrix effect and correction by standard addition in quantitative liquid chromatographic-mass spectrometric analysis of diarrhetic shellfish poisoning toxins","volume":"943","author":"Ito","year":"2002","journal-title":"J. Chromatogr. A"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"682","DOI":"10.1021\/jf034853x","article-title":"Quantitative determination of allicin in garlic: Supercritical fluid extraction and standard addition of alliin","volume":"52","author":"Rybak","year":"2004","journal-title":"J. Agric. Food Chem."},{"key":"ref_43","first-page":"319","article-title":"Analysis of 81 pesticides and metabolite residues in fruits and vegetables by diatomaceous earth column extraction and LC\/MS\/MS determination","volume":"17","author":"Tseng","year":"2009","journal-title":"J. Food Drug Anal."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.trac.2016.12.013","article-title":"The standard addition method revisited","volume":"89","author":"Andersen","year":"2017","journal-title":"Trends Anal. Chem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.jchromb.2012.10.011","article-title":"Analysis of new potential anticonvulsant compounds in mice brain tissue by SPE\/HPLC\/DAD","volume":"909","author":"Flieger","year":"2012","journal-title":"J. Chromatogr. B"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1080\/00032719.2014.938344","article-title":"Determination of 5-(3-Chlorophenyl)-4-hexyl-2,4-dihydro-3H-1,2,4-triazole-3-thione in Mouse Brain Tissue by Microwave-Assisted Extraction and High-Performance Liquid Chromatography with Fluorescence Detection","volume":"48","author":"Flieger","year":"2015","journal-title":"Anal. Lett."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1124\/jpet.103.060137","article-title":"Developmental expression of human hepatic CYP2C9 and CYP2C19","volume":"308","author":"Koukouritaki","year":"2004","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1111\/j.1365-2125.2009.03578.x","article-title":"Pharmacogenetics of CYP2C19: Functional and clinical implications of a new variant CYP2C19*17","volume":"69","author":"Girard","year":"2010","journal-title":"Br. J. Clin. Pharm."},{"key":"ref_49","unstructured":"ICH, Q2(R1) (2005). Validation of Analytical Procedures: Text and Methodology, International Conference on Harmonization."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/7\/3358\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:40:51Z","timestamp":1760161251000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/7\/3358"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,3,25]]},"references-count":49,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2021,4]]}},"alternative-id":["ijms22073358"],"URL":"https:\/\/doi.org\/10.3390\/ijms22073358","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2021,3,25]]}}}