Metabolism Study of Anamorelin, a GHSR1a Receptor Agonist Potentially Misused in Sport, with Human Hepatocytes and LC-HRMS/MS.
Prince Sellase GameliOmayema TaoussiGiuseppe BasileFrancesco P BusardòFrancesco Paolo BusardòPublished in: Metabolites (2023)
Anamorelin, developed for the treatment of cancer cachexia, is an orally active medication that improves appetite and food intake, thereby increasing body mass and physical functioning. It is classified as a growth hormone secretagogue and strictly monitored by the World Anti-Doping Agency (WADA), owing to its anabolic enhancing potential. Identifying anamorelin and/or metabolite biomarkers of consumption is critical in doping controls. However, there are currently no data available on anamorelin human metabolic fate. The aim of this study was to investigate and identify biomarkers characteristic of anamorelin intake using in silico metabolite predictions with GLORYx, in vitro incubation with 10-donor-pooled human hepatocytes, liquid chromatography-high-resolution tandem mass spectrometry (LC-HRMS/MS) analysis, and data processing with Thermo Scientific's Compound Discoverer. In silico prediction resulted in N -acetylation at the methylalanyl group as the main transformation (score, 88%). Others including hydroxylation at the indole substructure, and oxidation and N -demethylation at the trimethylhydrazino group were predicted (score, ≤36%). Hepatocyte incubations resulted in 14 phase I metabolites formed through N -demethylation at the trimethylhydrazino group, N -dealkylation at the piperidine ring, and oxidation at the indole and methylalanyl groups; and two phase II glucuronide conjugates occurring at the indole. We propose four metabolites detected as specific biomarkers for toxicological screening.
Keyphrases
- liquid chromatography
- tandem mass spectrometry
- mass spectrometry
- high resolution mass spectrometry
- endothelial cells
- ultra high performance liquid chromatography
- high resolution
- simultaneous determination
- ms ms
- growth hormone
- phase ii
- high performance liquid chromatography
- clinical trial
- induced pluripotent stem cells
- gas chromatography
- pluripotent stem cells
- solid phase extraction
- multiple sclerosis
- emergency department
- molecular docking
- liver injury
- healthcare
- open label
- risk assessment
- randomized controlled trial
- phase iii
- papillary thyroid
- big data
- study protocol
- drug induced
- double blind
- histone deacetylase
- artificial intelligence
- squamous cell