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Abstract
Evobrutinib is a highly selective, covalent, central nervous system-penetrant Bruton’s tyrosine kinase (BTK) inhibitor, currently in Phase III trials for the treatment of relapsing multiple sclerosis. One major circulating metabolite of evobrutinib has been previously identified as the racemic dihydro-diol M463-2 (MSC2430422) in a Phase I human mass balance study.
Phenotyping experiments were conducted to confirm the metabolic pathway of evobrutinib to M463-2. Ratio of the enantiomers was determined by enantioselective liquid chromatography with tandem mass spectrometry analysis of plasma samples from humans and preclinical species. Drug-drug interaction (DDI) characterisation, evaluation of pharmacological activity on BTK, and off-target screening experiments followed assessing safety of the metabolite.
The biotransformation of evobrutinib to M463-2 was determined to be a two-step process with a CYP-mediated oxidation acting to form an epoxide intermediate, which was further hydrolysed by soluble and mitochondrial epoxide hydrolase. Only the (S)-enantiomer was determined to be a major metabolite, the (R)-enantiomer was minor. In vitro studies demonstrated the (S)-enantiomer lacked clinically relevant pharmacological activity, off-target effects and DDIs.
The biotransformation of evobrutinib to its major metabolite has been elucidated, with the major (S)-enantiomer being shown to pose no on/off target or DDI risks.
Acknowledgments
Merck Healthcare KGaA, Darmstadt, Germany was involved in the experimental design, collection, analysis, and interpretation of the data, and the development of this manuscript. Medical writing and editorial support were provided by Jack Lochray of Bioscript Group Ltd, Macclesfield, UK, and supported by Merck Healthcare KGaA, Darmstadt, Germany (CrossRef Funder ID: 10.13039/100009945). The authors would like to thank Karthik Venkatakrishnan for reviewing the manuscript and providing valuable input, Birikiti Kidane for conducting some of the phenotyping experiments, Hammock Lab from University of California Davis for supporting with the epoxide hydrolase enzymes, Ursula Boschert and Yasemin Begum Alankus for the support and monitoring of the kinase profiling study, and Maximillian Kranz and Matthias Bader for supporting with the metabolite profiling and identification studies.
Disclosure statement
H Scheible, H Schieferstein, K Pusecker, U Gradhand, S Gopalakrishnan, K Iqbal and K Georgi are or were employees of Merck Healthcare KGaA, Darmstadt, Germany.
R Schmidt, J Dong, R Jones, J Bolleddula and M Dyroff are or were employees of EMD Serono Research & Development Institute, Inc., Billerica, MA, USA, an affiliate of Merck KGaA. C Meli is an employee of Merck Ltd., Piedmont, Italy, an affiliate of Merck KGaA.
Data availability statement
Data are available upon reasonable request. Any requests for data by qualified scientific and medical researchers for legitimate research purposes will be subject to Merck Healthcare KGaA, Darmstadt, Germany’s Data Sharing Policy. All requests should be submitted in writing to Merck Healthcare KGaA, Darmstadt, Germany’s data sharing portal https://www.merckgroup.com/en/research/our-approach-to-research-and-development/healthcare/clinical-trials/commitment-responsible-data-sharing.html. When Merck Healthcare KGaA, Darmstadt, Germany has a co-research, co-development, or co-marketing or co-promotion agreement, or when the product has been out-licensed, the responsibility for disclosure might be dependent on the agreement between parties. Under these circumstances, Merck Healthcare KGaA, Darmstadt, Germany will endeavour to gain agreement to share data in response to requests.