title: Mitigation of reactive metabolite formation for a series of 3-amino-2-pyridone inhibitors of Bruton's tyrosine kinase (BTK).
abstract: Reactive metabolites have been putatively linked to many adverse drug reactions including idiosyncratic toxicities for a number of drugs with black box warnings or withdrawn from the market. Therefore, it is desirable to minimize the risk of reactive metabolite formation for lead molecules in optimization, in particular for non-life threatening chronic disease, to maximize benefit to risk ratio. This article describes our effort in addressing reactive metabolite issues for a series of 3-amino-2-pyridone inhibitors of BTK, e.g. compound 1 has a value of 459pmol/mg protein in the microsomal covalent binding assay. Parallel approaches were taken to successfully resolve the issues: establishment of a predictive screening assay with correlation association of covalent binding assay, identification of the origin of reactive metabolite formation using MS/MS analysis of HLM as well as isolation and characterization of GSH adducts. This ultimately led to the discovery of compound 7 (RN941) with significantly reduced covalent binding of 26pmol/mg protein.
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States. Electronic address: yan.lou@gmail.com.
authors.forename: Yan
authors.initials: Y
authors.lastname: Lou
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Francisco
authors.initials: F
authors.lastname: Lopez
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Yongying
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Xiaochun
authors.initials: X
authors.lastname: Han
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Chris
authors.initials: C
authors.lastname: Brotherton
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Roland
authors.initials: R
authors.lastname: Billedeau
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Steve
authors.initials: S
authors.lastname: Gabriel
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Shelly
authors.initials: S
authors.lastname: Gleason
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: David M
authors.initials: DM
authors.lastname: Goldstein
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Ramona
authors.initials: R
authors.lastname: Hilgenkamp
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Buelent
authors.initials: B
authors.lastname: Kocer
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Lucja
authors.initials: L
authors.lastname: Orzechowski
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Jenny
authors.initials: J
authors.lastname: Tan
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Peter
authors.initials: P
authors.lastname: Wovkulich
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Bo
authors.initials: B
authors.lastname: Wen
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: David
authors.initials: D
authors.lastname: Fry
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Paola
authors.initials: P
authors.lastname: Di Lello
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Lucy
authors.initials: L
authors.lastname: Chen
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Fang-Jie
authors.initials: FJ
authors.lastname: Zhang
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Fretland
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Anjali
authors.initials: A
authors.lastname: Nangia
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Tian
authors.initials: T
authors.lastname: Yang
authors.affiliation: Hoffmann-La Roche Ltd., pRED, Pharma Research & Early Development, 340 Kingsland Street, Nutley 07110, United States.
authors.forename: Timothy D
authors.initials: TD
authors.lastname: Owens
mesh_terms: D000077329:Agammaglobulinaemia Tyrosine Kinase; D005978:Glutathione / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D008861:Microsomes / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011728:Pyridones / Q000737:chemistry* / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D047428:Protein Kinase Inhibitors; D011728:Pyridones; D011505:Protein-Tyrosine Kinases; D000077329:Agammaglobulinaemia Tyrosine Kinase; D005978:Glutathione
keywords: BTK; Bruton’s tyrosine kinase; Covalent binding; Glutathione adduct; Reactive metabolite