Investigation of Bemethyl Biotransformation Pathways by Combination of LC–MS/HRMS and In Silico Methods
Bemethyl is an actoprotector, an antihypoxant, and a moderate psychostimulant. Even though the therapeutic effectiveness of bemethyl is well documented, there is a gap in knowledge regarding its metabolic products and their quantitative and qualitative characteristics. Since 2018, bemethyl is includ...
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doaj-b9c94e55f1a3473684752a65c31dcbb72021-08-26T13:53:40ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-08-01229021902110.3390/ijms22169021Investigation of Bemethyl Biotransformation Pathways by Combination of LC–MS/HRMS and In Silico MethodsDaria A. Belinskaia0Elena I. Savelieva1Georgy V. Karakashev2Olga I. Orlova3Mikhail A. Leninskii4Nataliia S. Khlebnikova5Natalia N. Shestakova6Alexandra R. Kiskina7Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, Pr. Torez 44, 194223 St. Petersburg, RussiaSechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, Pr. Torez 44, 194223 St. Petersburg, RussiaSechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, Pr. Torez 44, 194223 St. Petersburg, RussiaSechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, Pr. Torez 44, 194223 St. Petersburg, RussiaSechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, Pr. Torez 44, 194223 St. Petersburg, RussiaSechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, Pr. Torez 44, 194223 St. Petersburg, RussiaSechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, Pr. Torez 44, 194223 St. Petersburg, RussiaSechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, Pr. Torez 44, 194223 St. Petersburg, RussiaBemethyl is an actoprotector, an antihypoxant, and a moderate psychostimulant. Even though the therapeutic effectiveness of bemethyl is well documented, there is a gap in knowledge regarding its metabolic products and their quantitative and qualitative characteristics. Since 2018, bemethyl is included to the Monitoring Program of the World Anti-Doping Agency, which highlights the challenge of identifying its urinary metabolites. The objective of the study was to investigate the biotransformation pathways of bemethyl using a combination of liquid chromatography-high-resolution mass spectrometry and in silico studies. Metabolites were analyzed in a 24 h rat urine collected after oral administration of bemethyl at a single dose of 330 mg/kg. The urine samples were prepared for analysis by a procedure developed in the present work and analyzed by high performance liquid chromatography–tandem mass spectrometry. For the first time, nine metabolites of bemethyl with six molecular formulas were identified in rat urine. The most abundant metabolite was a benzimidazole–acetylcysteine conjugate; this biotransformation pathway is associated with the detoxification of xenobiotics. The BioTransformer and GLORY computational tools were used to predict bemethyl metabolites in silico. The molecular docking of bemethyl and its derivatives to the binding site of glutathione S-transferase has revealed the mechanism of bemethyl conjugation with glutathione. The findings will help to understand the pharmacokinetics and pharmacodynamics of actoprotectors and to improve antihypoxant and adaptogenic therapy.https://www.mdpi.com/1422-0067/22/16/9021bemethyl2-(ethylthio)benzimidazoleactoprotectordopingrat urinemetabolite |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Daria A. Belinskaia Elena I. Savelieva Georgy V. Karakashev Olga I. Orlova Mikhail A. Leninskii Nataliia S. Khlebnikova Natalia N. Shestakova Alexandra R. Kiskina |
spellingShingle |
Daria A. Belinskaia Elena I. Savelieva Georgy V. Karakashev Olga I. Orlova Mikhail A. Leninskii Nataliia S. Khlebnikova Natalia N. Shestakova Alexandra R. Kiskina Investigation of Bemethyl Biotransformation Pathways by Combination of LC–MS/HRMS and In Silico Methods International Journal of Molecular Sciences bemethyl 2-(ethylthio)benzimidazole actoprotector doping rat urine metabolite |
author_facet |
Daria A. Belinskaia Elena I. Savelieva Georgy V. Karakashev Olga I. Orlova Mikhail A. Leninskii Nataliia S. Khlebnikova Natalia N. Shestakova Alexandra R. Kiskina |
author_sort |
Daria A. Belinskaia |
title |
Investigation of Bemethyl Biotransformation Pathways by Combination of LC–MS/HRMS and In Silico Methods |
title_short |
Investigation of Bemethyl Biotransformation Pathways by Combination of LC–MS/HRMS and In Silico Methods |
title_full |
Investigation of Bemethyl Biotransformation Pathways by Combination of LC–MS/HRMS and In Silico Methods |
title_fullStr |
Investigation of Bemethyl Biotransformation Pathways by Combination of LC–MS/HRMS and In Silico Methods |
title_full_unstemmed |
Investigation of Bemethyl Biotransformation Pathways by Combination of LC–MS/HRMS and In Silico Methods |
title_sort |
investigation of bemethyl biotransformation pathways by combination of lc–ms/hrms and in silico methods |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1661-6596 1422-0067 |
publishDate |
2021-08-01 |
description |
Bemethyl is an actoprotector, an antihypoxant, and a moderate psychostimulant. Even though the therapeutic effectiveness of bemethyl is well documented, there is a gap in knowledge regarding its metabolic products and their quantitative and qualitative characteristics. Since 2018, bemethyl is included to the Monitoring Program of the World Anti-Doping Agency, which highlights the challenge of identifying its urinary metabolites. The objective of the study was to investigate the biotransformation pathways of bemethyl using a combination of liquid chromatography-high-resolution mass spectrometry and in silico studies. Metabolites were analyzed in a 24 h rat urine collected after oral administration of bemethyl at a single dose of 330 mg/kg. The urine samples were prepared for analysis by a procedure developed in the present work and analyzed by high performance liquid chromatography–tandem mass spectrometry. For the first time, nine metabolites of bemethyl with six molecular formulas were identified in rat urine. The most abundant metabolite was a benzimidazole–acetylcysteine conjugate; this biotransformation pathway is associated with the detoxification of xenobiotics. The BioTransformer and GLORY computational tools were used to predict bemethyl metabolites in silico. The molecular docking of bemethyl and its derivatives to the binding site of glutathione S-transferase has revealed the mechanism of bemethyl conjugation with glutathione. The findings will help to understand the pharmacokinetics and pharmacodynamics of actoprotectors and to improve antihypoxant and adaptogenic therapy. |
topic |
bemethyl 2-(ethylthio)benzimidazole actoprotector doping rat urine metabolite |
url |
https://www.mdpi.com/1422-0067/22/16/9021 |
work_keys_str_mv |
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