Login Register Cart 0
Generic placeholder image

Drug Metabolism and Bioanalysis Letters

Editor-in-Chief

ISSN (Print): 2949-6810
ISSN (Online): 2949-6829

Back
Journal Home About Journal Editorial Board Current Issue Volumes/Issues
Subscribe
Research Article

Species-specific Bioactivation of Morpholines as a Causative of Drug Induced Liver Injury Observed in Monkeys

Author(s): Mithat Gunduz*, Upendra A. Argikar, Amanda L. Cirello, Alan P. Brown, Simone Bonazzi and Markus Walles

Volume 17, Issue 1, 2024

Published on: 01 December, 2023

Page: [13 - 22] Pages: 10

DOI: 10.2174/0118723128260455231104180653

Price: $65

Abstract

Background: Everolimus, an allosteric mechanistic target of rapamycin (mTOR) inhibitor, recently demonstrated the therapeutic value of mTOR inhibitors for Central Nervous System (CNS) indications driven by hyperactivation of mTOR. A newer, potent brain-penetrant analog of everolimus, referred to as (1) in this manuscript [(S)-3-methyl-4-(7-((R)-3-methylmorpholino)-2- (thiazol-4-yl)-3H-imidazo[4,5-b]pyridin-5-yl)morpholine,(1)] catalytically inhibits mTOR function in the brain and increases the lifespan of mice with neuronal mTOR hyperactivation.

Introduction: Early evaluation of the safety of 1 was conducted in cynomolgus monkeys in which oral doses were administered to three animals in a rising-dose fashion (from 2 to 30 mg/kg/day). 1 produced severe toxicity including the evidence of hepatic toxicity, along with non-dose proportional increases in drug exposure. Investigations of cross-species hepatic bioactivation of 1 were conducted to assess whether the formation of reactive drug metabolites was associated with the mechanism of liver toxicity.

Methods: 1 contained two morpholine rings known as structural alerts and can potentially form reactive intermediates through oxidative metabolism. Bioactivation of 1 was investigated in rat, human and monkey liver microsomes fortified with trapping agents such as methoxylamine or potassium cyanide.

Results: Our results suggest that bioactivation of the morpholine moieties to reactive intermediates may have been involved in the mechanism of liver toxicity observed with 1. Aldehyde intermediates trappable by methoxylamine were identified in rat and monkey liver microsomal studies. In addition, a total of four cyano conjugates arising from the formation of iminium ion intermediates were observed and identified. These findings may potentially explain the observed monkey toxicity. Interestingly, methoxylamine or cyano adducts of 1 were not observed in human liver microsomes.

Conclusion: The bioactivation of 1 appears to be species-specific. Circumstantial evidence for the toxicity derived from 1 point to the formation of iminium ion intermediates trappable by cyanide in monkey liver microsomes. The cyano conjugates were only observed in monkey liver microsomes, potentially pointing to cause at least the hepatotoxicity observed in monkeys. In contrast, methoxylamine conjugates were detected in both rat and monkey liver microsomes, with only a trace amount in human liver microsomes. Cyano conjugates were not observed in human liver microsomes, challenging the team on the drugability and progressivity of 1 through drug development. The mechanisms for drug-induced liver toxicity are multifactorial. These results are highly suggestive that the iminium ion may be an important component in the mechanism of liver toxicity 1 observed in the monkey.

Keywords: Potassium cyanide, iminium ion intermediate, methoxylamine, aldehyde intermediate, liver microsomes, bioactivation, morpholine.

« Previous Next »
Graphical Abstract

[1]
Ballou, L.M.; Lin, R.Z. Rapamycin and mTOR kinase inhibitors. J. Chem. Biol., 2008, 1(1-4), 27-36.
[http://dx.doi.org/10.1007/s12154-008-0003-5] [PMID: 19568796]
[2]
Zheng, Y.; Jiang, Y. mTOR inhibitors at a glance. Mol. Cell. Pharmacol., 2015, 7(2), 15-20.
[PMID: 27134695]
[3]
Borsari, C.; Keles, E.; Rageot, D.; Treyer, A.; Bohnacker, T.; Bissegger, L.; De Pascale, M.; Melone, A.; Sriramaratnam, R.; Beaufils, F.; Hamburger, M.; Hebeisen, P.; Löscher, W.; Fabbro, D.; Hillmann, P.; Wymann, M.P. 4-(Difluoromethyl)-5-(4-((3 R, 5 S)-3,5-dimethylmorpholino)-6-((R)-3-methylmorpholino)-1,3,5-triazin-2-yl)pyridin-2-amine (PQR626), a potent, prally available, and brain-penetrant mTOR inhibitor for the treatment of neurological disorders. J. Med. Chem., 2020, 63(22), 13595-13617.
[http://dx.doi.org/10.1021/acs.jmedchem.0c00620] [PMID: 33166139]
[4]
Han, X.; Goh, K.Y.; Lee, W.X.; Choy, S.M.; Tang, H.W. The importance of mTORC1-autophagy axis for skeletal muscle diseases. Int. J. Mol. Sci., 2022, 24(1), 297.
[http://dx.doi.org/10.3390/ijms24010297 ] [PMID: 36613741]
[5]
Bushee, J.L.; Argikar, U.A. An experimental approach to enhance precursor ion fragmentation for metabolite identification studies: application of dual collision cells in an orbital trap. Rapid Commun. Mass Spectrom., 2011, 25(10), 1356-1362.
[http://dx.doi.org/10.1002/rcm.4996] [PMID: 21504000]
[6]
Gunduz, M.; Argikar, U.A.; Baeschlin, D.; Ferreira, S.; Hosagrahara, V.; Harriman, S. Identification of a novel N-carbamoyl glucuronide: In vitro, in vivo, and mechanistic studies. Drug Metab. Dispos., 2010, 38(3), 361-367.
[http://dx.doi.org/10.1124/dmd.109.030650] [PMID: 20008038]
[7]
Andrade, R.J.; Chalasani, N.; Björnsson, E.S.; Suzuki, A.; Kullak-Ublick, G.A.; Watkins, P.B.; Devarbhavi, H.; Merz, M.; Lucena, M.I.; Kaplowitz, N.; Aithal, G.P. Drug-induced liver injury. Nat. Rev. Dis. Primers, 2019, 5(1), 58.
[http://dx.doi.org/10.1038/s41572-019-0105-0] [PMID: 31439850]
[8]
Stepan, A.F.; Walker, D.P.; Bauman, J.; Price, D.A.; Baillie, T.A.; Kalgutkar, A.S.; Aleo, M.D. Structural alert/reactive metabolite concept as applied in medicinal chemistry to mitigate the risk of idiosyncratic drug toxicity: A perspective based on the critical examination of trends in the top 200 drugs marketed in the United States. Chem. Res. Toxicol., 2011, 24(9), 1345-1410.
[http://dx.doi.org/10.1021/tx200168d] [PMID: 21702456]
[9]
Argikar, U.A.; Mangold, J.B.; Harriman, S.P. Strategies and chemical design approaches to reduce the potential for formation of reactive metabolic species. Curr. Top. Med. Chem., 2011, 11(4), 419-449.
[http://dx.doi.org/10.2174/156802611794480891] [PMID: 21320068]
[10]
Kalgutkar, A.S.; Dalvie, D. Predicting toxicities of reactive metabolite-positive drug candidates. Annu. Rev. Pharmacol. Toxicol., 2015, 55(1), 35-54.
[http://dx.doi.org/10.1146/annurev-pharmtox-010814-124720] [PMID: 25292426]
[11]
Kalgutkar, A.S.; Didiuk, M.T. Structural alerts, reactive metabolites, and protein covalent binding: How reliable are these attributes as predictors of drug toxicity? Chem. Biodivers., 2009, 6(11), 2115-2137.
[http://dx.doi.org/10.1002/cbdv.200900055] [PMID: 19937848]
[12]
Kalgutkar, A.S. Liabilities associated with the formation of “hard” electrophiles in reactive metabolite trapping screens. Chem. Res. Toxicol., 2017, 30(1), 220-238.
[http://dx.doi.org/10.1021/acs.chemrestox.6b00332] [PMID: 27802597]
[13]
Bergström, M.A.; Isin, E.M.; Castagnoli, N., Jr; Milne, C.E. Bioactivation pathways of the cannabinoid receptor 1 antagonist rimonabant. Drug Metab. Dispos., 2011, 39(10), 1823-1832.
[http://dx.doi.org/10.1124/dmd.111.039412] [PMID: 21733882]
[14]
Zheng, J.; Xin, Y.; Zhang, J.; Subramanian, R.; Murray, B.P.; Whitney, J.A.; Warr, M.R.; Ling, J.; Moorehead, L.; Kwan, E.; Hemenway, J.; Smith, B.J.; Silverman, J.A. Pharmacokinetics and disposition of momelotinib revealed a disproportionate human metabolite-resolution for clinical development. Drug Metab. Dispos., 2018, 46(3), 237-247.
[http://dx.doi.org/10.1124/dmd.117.078899] [PMID: 29311136]
[15]
Bolleddula, J.; DeMent, K.; Driscoll, J.P.; Worboys, P.; Brassil, P.J.; Bourdet, D.L. Biotransformation and bioactivation reactions of alicyclic amines in drug molecules. Drug Metab. Rev., 2014, 46(3), 379-419.
[http://dx.doi.org/10.3109/03602532.2014.924962] [PMID: 24909234]
[16]
Lau, D.H.M.; Lewis, A.D.; Sikic, B.I. Association of DNA cross-linking with potentiation of the morpholino derivative of doxorubicin by human liver microsomes. J. Natl. Cancer Inst., 1989, 81(13), 1034-1038.
[http://dx.doi.org/10.1093/jnci/81.13.1034] [PMID: 2733045]
[17]
Gunduz, M.; Cirello, A.L.; Klimko, P.; Dumouchel, J.L.; Argikar, U.A. Genotoxicity of 4-(piperazin-1-yl)-8-(trifluoromethyl)pyrido[2,3-e][1,2,4] triazolo[4,3-a]pyrazine, a potent H4 receptor antagonist for the treatment of allergy: Evidence of glyoxal intermediate involvement. Drug Metab. Lett., 2017, 11(2), 144-148.
[PMID: 29110630]
[18]
Kenna, J.G.; Taskar, K.S.; Battista, C.; Bourdet, D.L.; Brouwer, K.L.R.; Brouwer, K.R.; Dai, D.; Funk, C.; Hafey, M.J.; Lai, Y.; Maher, J.; Pak, Y.A.; Pedersen, J.M.; Polli, J.W.; Rodrigues, A.D.; Watkins, P.B.; Yang, K.; Yucha, R.W. Can bile salt export pump inhibition testing in drug discovery and development reduce liver injury risk? An international transporter consortium perspective. Clin. Pharmacol. Ther., 2018, 104(5), 916-932.
[http://dx.doi.org/10.1002/cpt.1222] [PMID: 30137645]
[19]
Whitebread, S.; Fekete, A.; Jin, H.; Armstrong, D.; Urban, L. Inhibition of bile salt export pump (BSEP) in relation to systemic exposure: A risk factor for drug induced liver injury (DILI). J. Pharmacol. Toxicol. Methods, 2017, 88, 215.
[http://dx.doi.org/10.1016/j.vascn.2017.09.154]

Rights & Permissions Print Cite
Article Metrics
98
2
© 2024 Bentham Science Publishers | Privacy Policy