title: Metabolite identification of bentysrepinine (Y101), a novel anti-HBV agent in rats using a five-step strategy based on a combined workflow with two different platforms of liquid chromatography-tandem mass spectrometry.
abstract: Bentysrepinine (Y101), a derivative of repensine (a compound isolated from Dichondrarepens Forst), is a novel phenyalanine dipeptide inhibiting DNA-HBV and cccDNA activities and is currently under development for the treatment of hepatitis B virus (HBV)-infected hepatitis. Our previous study implied that there might be an existence of extensive metabolism of Y101 in rats. Therefore, it is necessary to perform metabolic profiling study to further evaluate its safety and drug-like properties. In this study, the metabolism of Y101 in rats was investigated by a convincible five-step strategy to characterize metabolites in plasma and that excreted into urine, bile and feces. The five-step strategy was realized by using an combined workflow on two different MS platforms, including various scan modes of liquid chromatography with hybrid quadruple-linear ion trap mass spectrometry (LC-QTRAP-MS/MS) and various post-acquiring data mining tools of liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (LC-QTOF-MS/MS). QTOF MS/MS was employed as a powerful complementary tool to enable high confidence of metabolites identification using its functions of accurate MS and MS/MS fragmentation. As a result, a total of 30 metabolites were detected, including 25 phase I and 5 phase II metabolites. Among them, four primary metabolites (M6-M9) were further identified by comparing with the authentic standards chemically synthesized. The possible metabolic pathways of Y101 in rats were proposed to be amide hydrolysis, monohydroxylation, dihydroxylation, N-oxidation, demethylation, methylation, glucosidation and glucuronidation. This is the first study of the metabolism of Y101 in rats. The five-step strategy was successfully used to systematically characterize metabolites of Y101 in rats, and it would be generally applied for metabolite identification of new drug candidate.
authors.affiliation: Tianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine, Institute of Radiation Medicine, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300192, China. Electronic address: fanhr99@163.com.
authors.forename: Huirong
authors.initials: H
authors.lastname: Fan
authors.affiliation: The Key Laboratory of Chemistry for Natural Products of Guizhou Province and Chinese Academy of Sciences, Guiyang 550002, China.
authors.forename: Zhanxing
authors.initials: Z
authors.lastname: Hu
authors.affiliation: WuXi AppTec Inc., Shanghai 200131, China.
authors.forename: Ruixing
authors.initials: R
authors.lastname: Li
authors.affiliation: Tianjin University, Tianjin 300072, China.
authors.forename: Shiqi
authors.initials: S
authors.lastname: Dong
authors.affiliation: State Key Laboratory of Drug Delivery Technology and Pharmacokinetics, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Gu
authors.affiliation: Shanghai AB Sciex Analytical Instrument Trading Co. Ltd, Shanghai 200335, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Drug Delivery Technology and Pharmacokinetics, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China.
authors.forename: Duanyun
authors.initials: D
authors.lastname: Si
authors.affiliation: The Key Laboratory of Chemistry for Natural Products of Guizhou Province and Chinese Academy of Sciences, Guiyang 550002, China.
authors.forename: Guangyi
authors.initials: G
authors.lastname: Liang
authors.affiliation: State Key Laboratory of Drug Delivery Technology and Pharmacokinetics, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China. Electronic address: liuchangxiao@163.com.
authors.forename: Changxiao
authors.initials: C
authors.lastname: Liu
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000032:analysis / Q000378:metabolism*; D001549:Benzamides / Q000032:analysis / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004151:Dipeptides / Q000032:analysis / Q000378:metabolism*; D005260:Female; D006509:Hepatitis B / Q000188:drug therapy; D006515:Hepatitis B virus / Q000187:drug effects; D008297:Male; D053858:Metabolic Networks and Pathways; D051381:Rats; D017207:Rats, Sprague-Dawley; D053719:Tandem Mass Spectrometry / Q000379:methods*; D057188:Workflow
chemical_list: D000998:Antiviral Agents; D001549:Benzamides; D004151:Dipeptides; C573540:N-(N-benzoyl-O-(2-dimethylaminoethyl)tyrosyl)phenylalaninol
keywords: Five-step strategy; Identification; LC-QTOF-MS/MS; LC-QTRAP-MS/MS; Metabolite; Y101