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In Vitro Metabolism by Aldehyde Oxidase Leads to Poor Pharmacokinetic Profile in Rats for c-Met Inhibitor MET401

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Abstract

Background and Objectives

MET401 is a potent and selective c-Met inhibitor with a novel triazolopyrimidine scaffold. The aim of this study was to determine the pharmacokinetic profile of MET401 in preclinical species, and to identify the metabolic soft spot and enzyme involved, in order to help medicinal chemists to modify the compound to improve the pharmacokinetic profile.

Methods

A metabolite identification study was performed in different liver fractions from various species. Chemical inhibition with selective cytochrome P450 (CYP) and molybdenum hydroxylase inhibitors was carried out to identify the enzyme involved. The deuterium substitution strategy was adopted to reduce metabolism. Pharmacokinetic studies were performed in rats to confirm the effect.

Results

Although M-2 is a minor metabolite in liver microsomal incubations, it became the predominant metabolite in incubations with liver S9, cytosol, hepatocytes and rat pharmacokinetic study. M-2 was synthesized enzymatically and the structure was identified as a mono-oxidation on the triazolopyrimidine moiety. The M-2 formation was ascribed to aldehyde oxidase (AO)-mediated metabolism based on the following evidence—M-2 production was NADPH independent, pan-CYP inhibitor 1-aminobenzotriazole and xanthine oxidase inhibitor allopurinol did not inhibit M-2 formation, and AO inhibitors menadione and raloxifene inhibited M-2 formation. The deuterated analog MET763 demonstrated an improved pharmacokinetic profile with lower clearance, longer terminal half-life and double oral exposure compared with MET401 in rats.

Conclusions

These results indicate that the main metabolic pathway of MET401 is AO-mediated metabolism, which leads to poor in vivo pharmacokinetic profiles in rodents. The deuterium substitution strategy could be used to reduce AO-mediated metabolism liability.

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References

  1. Cañadas I, Rojo F, Arumí-Uría M, Rovira A, Albanell J, Arriola E. C-MET as a new therapeutic target for the development of novel anticancer drugs. Clin Transl Oncol. 2010;12:253–60.

    Article  CAS  PubMed  Google Scholar 

  2. Liu X, Newton RC, Scherle PA. Developing c-MET pathway inhibitors for cancer therapy: progress and challenges. Trends Mol Med. 2010;16:37–45.

    Article  CAS  PubMed  Google Scholar 

  3. Parikh PK, Ghate MD. Recent advances in the discovery of small molecule c-Met Kinase inhibitors. Eur J Med Chem. 2018;143:1103–38.

    Article  CAS  PubMed  Google Scholar 

  4. Comoglio PM, Trusolino L, Boccaccio C. Known and novel roles of the MET oncogene in cancer: a coherent approach to targeted therapy. Nat Rev Cancer. 2018;18:341–58.

    Article  CAS  PubMed  Google Scholar 

  5. Cui JJ. Targeting receptor tyrosine kinase MET in cancer: small molecule inhibitors and clinical progress. J Med Chem. 2014;57:4427–53.

    Article  CAS  PubMed  Google Scholar 

  6. Tiedt R, Degenkolbe E, Furet P, Appleton BA, Wagner S, Schoepfer J, et al. A drug resistance screen using a selective MET inhibitor reveals a spectrum of mutations that partially overlap with activating mutations found in cancer patients. Cancer Res. 2011;71:5255–64.

    Article  CAS  PubMed  Google Scholar 

  7. Zhan Z, Peng X, Liu Q, Chen F, Ji Y, Yao S, et al. Discovery of 6-(difluoro(6-(4-fluorophenyl)-[1,2,4]triazolo[4,3-b][1,2,4]triazin-3-yl)methyl)quinoline as a highly potent and selective c-Met inhibitor. Eur J Med Chem. 2016;116:239–51.

    Article  CAS  PubMed  Google Scholar 

  8. Zhao F, Zhang J, Zhang L, Hao Y, Shi C, Xia G, et al. Discovery and optimization of a series of imidazo[4,5-b]pyrazine derivatives as highly potent and exquisitely selective inhibitors of the mesenchymal–epithelial transition factor (c-Met) protein kinase. Bioorg Med Chem. 2016;24:4281–90.

    Article  CAS  PubMed  Google Scholar 

  9. Ye L, Wu J, Yang J, Chen W, Luo Y, Zhang Y. Design, synthesis and molecular docking analysis of some novel 7-[(quinolin-6-yl)methyl] purines as potential c-Met inhibitors. Med Chem Res. 2015;24:3327–33.

    Article  CAS  Google Scholar 

  10. Ye L, Ou X, Tian Y, Yu B, Luo Y, Feng B, et al. Indazoles as potential c-met inhibitors: design, synthesis and molecular docking studies. Eur J Med Chem. 2013;65:112–8.

    Article  CAS  PubMed  Google Scholar 

  11. Jia H, Dai G, Weng J, Zhang Z, Wang Q, Zhou F, et al. Discovery of (S)-1-(1-(Imidazo[1,2-a]pyridin-6-yl)ethyl)-6-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazine (volitinib) as a highly potent and selective mesenchymal-epithelial transition factor (c-Met) inhibitor in clinical development for tre. J Med Chem. 2014;57:7577–89.

    Article  CAS  PubMed  Google Scholar 

  12. Cui JJ, Shen H, Tran-Dubé M, Nambu M, McTigue M, Grodsky N, et al. Lessons from (S)-6-(1-(6-(1-methyl-1H-pyrazol-4-yl)-[1,2,4]triazolo[4,3-b]pyridazin-3-yl)ethyl)quinoline (PF-04254644), an inhibitor of receptor tyrosine kinase c-Met with high protein kinase selectivity but broad phosphodiesterase family inhibition leading to myocardial regeneration in rats. J Med Chem. 2013;56:6651–65.

    Article  CAS  PubMed  Google Scholar 

  13. Pirouzpanah S, Saieed P, Rashidi MR, Reza RM, Delazar A, Abbas D, et al. Inhibitory effects of Ruta graveolens L. extract on guinea pig liver aldehyde oxidase. Chem Pharm Bull (Tokyo). 2006;54:9–13.

    Article  Google Scholar 

  14. Kadam RS, Iyer KR. Isolation of liver aldehyde oxidase containing fractions from different animals and determination of kinetic parameters for benzaldehyde. Indian J Pharm Sci. 2008;70:85–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Linder CD, Renaud NA, Hutzler JM. Is 1-aminobenzotriazole an appropriate in vitro tool as a nonspecific cytochrome P450 inactivator? Drug Metab Dispos. 2009;37:10–3.

    Article  CAS  PubMed  Google Scholar 

  16. Bjornsson TD, Callaghan JT, Einolf HJ, Fischer V, Gan L, Grimm S, et al. The conduct of in vitro and in vivo drug-drug interaction studies: a PhRMA perspective. J Clin Pharmacol. 2003;43:443–69.

    Article  CAS  PubMed  Google Scholar 

  17. Johns DG. Human liver aldehyde oxidase: differential inhibition of oxidation of charged and uncharged substrates. J Clin Invest. 1967;46:1492–505.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Massey V, Komai H, Palmer G, Elion GB. On the mechanism of inactivation of xanthine oxidase by allopurinol and other pyrazolo[3,4-d]pyrimidines. J Biol Chem. 1970;245:2837–44.

    CAS  PubMed  Google Scholar 

  19. Sahi J, Khan KK, Black CB. Aldehyde oxidase activity and inhibition in hepatocytes and cytosolic fractions from mouse, rat, monkey and human. Drug Metab Lett. 2008;2:176–83.

    Article  CAS  PubMed  Google Scholar 

  20. Zhang JW, Xiao W, Gao ZT, Yu ZT, Zhang J-YJ. Metabolism of c-Met kinase inhibitors containing quinoline by aldehyde oxidase, electron donating, and steric hindrance effect. Drug Metab Dispos. 2018;46:1847–55.

    Article  CAS  PubMed  Google Scholar 

  21. Torres RA, Korzekwa KR, McMasters DR, Fandozzi CM, Jones JP. Use of density functional calculations to predict the regioselectivity of drugs and molecules metabolized by aldehyde oxidase. J Med Chem. 2007;50:4642–7.

    Article  CAS  PubMed  Google Scholar 

  22. Dalvie D, Sun H, Xiang C, Hu Q, Jiang Y, Kang P. Effect of structural variation on aldehyde oxidase-catalyzed oxidation of zoniporide. Drug Metab Dispos. 2012;40:1575–87.

    Article  CAS  PubMed  Google Scholar 

  23. Xu Y, Li L, Wang Y, Xing J, Zhou L, Zhong D, et al. Aldehyde oxidase mediated metabolism in drug-like molecules: a combined computational and experimental study. J Med Chem. 2017;60:2973–82.

    Article  CAS  PubMed  Google Scholar 

  24. Testa B, Krämer SD. The biochemistry of drug metabolism–an introduction: part 2. Redox reactions and their enzymes. Chem Biodivers. 2007;4:257–405.

    Article  CAS  PubMed  Google Scholar 

  25. Cleland WW. Isotope effects: determination of enzyme transition state structure. Methods Enzymol. 1995;249:341–73.

    Article  CAS  PubMed  Google Scholar 

  26. Alfaro JF, Joswig-Jones CA, Ouyang W, Nichols J, Crouch GJ, Jones JP. Purification and mechanism of human aldehyde oxidase expressed in Escherichia coli. Drug Metab Dispos. 2009;37:2393–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Shao L, Hewitt MC. The kinetic isotope effect in the search for deuterated drugs. Drug News Perspect. 2010;23:398–404.

    Article  CAS  PubMed  Google Scholar 

  28. Koniarczyk JL, Hesk D, Overgard A, Davies IW, McNally A. A general strategy for site-selective incorporation of deuterium and tritium into pyridines, diazines, and pharmaceuticals. J Am Chem Soc. 2018;140:1990–3.

    Article  CAS  PubMed  Google Scholar 

  29. Zhan Z, Peng X, Sun Y, Ai J, Duan W. Evaluation of deuterium-labeled JNJ38877605: pharmacokinetic, metabolic, and in vivo antitumor profiles. Chem Res Toxicol. 2018;31:1213–8.

    Article  CAS  PubMed  Google Scholar 

  30. Pryde DC, Dalvie D, Hu Q, Jones P, Obach RS, Tran TD. Aldehyde oxidase: an enzyme of emerging importance in drug discovery. J Med Chem. 2010;53:8441–60.

    Article  CAS  PubMed  Google Scholar 

  31. Kitamura S, Sugihara K, Ohta S. Drug-metabolizing ability of molybdenum hydroxylases. Drug Metab Pharmacokinet. 2006;21:83–98.

    Article  CAS  PubMed  Google Scholar 

  32. Lepri S, Ceccarelli M, Milani N, Tortorella S, Cucco A, Valeri A, et al. Structure–metabolism relationships in human-AOX: chemical insights from a large database of aza-aromatic and amide compounds. Proc Natl Acad Sci USA. 2017;114:E3178–87.

    Article  CAS  PubMed  Google Scholar 

  33. Sodhi JK, Wong S, Kirkpatrick DS, Liu L, Khojasteh SC, Hop CEC, et al. A novel reaction mediated by human aldehyde oxidase: amide hydrolysis of GDC-0834. Drug Metab Dispos. 2015;43:908–15.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Cruciani G, Milani N, Benedetti P, Lepri S, Cesarini L, Baroni M, et al. From experiments to a fast easy-to-use computational methodology to predict human aldehyde oxidase selectivity and metabolic reactions. J Med Chem. 2018;61:360–71.

    Article  CAS  PubMed  Google Scholar 

  35. Dittrich C, Greim G, Borner M, Weigang-Köhler K, Huisman H, Amelsberg A, et al. Phase I and pharmacokinetic study of BIBX 1382 BS, an epidermal growth factor receptor (EGFR) inhibitor, given in a continuous daily oral administration. Eur J Cancer. 2002;38:1072–80.

    Article  CAS  PubMed  Google Scholar 

  36. Zhang X, Liu H, Weller P, Zheng M, Tao W, Wang J, et al. In silico and in vitro pharmacogenetics: aldehyde oxidase rapidly metabolizes a p38 kinase inhibitor. Pharmacogenomics J. 2011;11:15–24.

    Article  CAS  PubMed  Google Scholar 

  37. Kaye B, Rance DJ, Waring L. Oxidative metabolism of carbazeran in vitro by liver cytosol of baboon and man. Xenobiotica. 1985;15:237–42.

    Article  CAS  PubMed  Google Scholar 

  38. Akabane T, Tanaka K, Irie M, Terashita S, Teramura T. Case report of extensive metabolism by aldehyde oxidase in humans: pharmacokinetics and metabolite profile of FK3453 in rats, dogs, and humans. Xenobiotica. 2011;41:372–84.

    Article  CAS  PubMed  Google Scholar 

  39. Calabrese CR, Loadman PM, Lim LS, Bibby MC, Double JA, Brown JE, et al. In vivo metabolism of the antitumor imidazoacridinone C1311 in the mouse and in vitro comparison with humans. Drug Metab Dispos. 1999;27:240–5.

    CAS  PubMed  Google Scholar 

  40. Schofield PC, Robertson IG, Paxton JW. Inter-species variation in the metabolism and inhibition of N-[(2′-dimethylamino)ethyl]acridine-4-carboxamide (DACA) by aldehyde oxidase. Biochem Pharmacol. 2000;59:161–5.

    Article  CAS  PubMed  Google Scholar 

  41. Fukiya K, Itoh K, Yamaguchi S, Kishiba A, Adachi M, Watanabe N, et al. A single amino acid substitution confers high cinchonidine oxidation activity comparable with that of rabbit to monkey aldehyde oxidase 1. Drug Metab Dispos. 2010;38:302–7.

    Article  CAS  PubMed  Google Scholar 

  42. Dick RA. Refinement of in vitro methods for identification of aldehyde oxidase substrates reveals metabolites of kinase inhibitors. Drug Metab Dispos. 2018;46:846–59.

    Article  CAS  PubMed  Google Scholar 

  43. Diamond S, Boer J, Maduskuie TP, Falahatpisheh N, Li Y, Yeleswaram S. Species-specific metabolism of SGX523 by aldehyde oxidase and the toxicological implications. Drug Metab Dispos. 2010;38:1277–85.

    Article  CAS  PubMed  Google Scholar 

  44. Lolkema MP, Bohets HH, Arkenau H-T, Lampo A, Barale E, de Jonge MJ, et al. The c-Met tyrosine kinase inhibitor JNJ-38877605 causes renal toxicity through species-specific insoluble metabolite formation. Clin Cancer Res. 2015;21:2297–304.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Zhao F, Zhang LD, Hao Y, Chen N, Bai R, Wang YJ, et al. Identification of 3-substituted-6-(1-(1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl)ethyl)quinoline derivatives as highly potent and selective mesenchymal-epithelial transition factor (c-Met) inhibitors via metabolite profiling-based structural optimization. Eur J Med Chem. 2017;134:147–58.

    Article  CAS  PubMed  Google Scholar 

  46. Su WG. Discovery, preclinical and early clinical evaluation of volitinib: a potent and selective c-Met kinase inhibitor. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research, April 5–9. San Diego: AACR; 2014. https://www.hmplglobal.com/en/publications/presentation-on-volitinib-hmpl-504-at-aacr-2014/

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Acknowledgements

We thank Novartis c-Met project team members, especially Global Discovery Chemistry Shanghai colleagues for their contributions.

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Author notes

  1. Jiang Wei Zhang

    Present address: InxMed, Shanghai, China

  2. Hai Bing Deng, Hong Ping Yu & Yao Chang Xu

    Present address: Abbisko Therapeutics, Shanghai, China

  3. Chun Ye Zhang

    Present address: Luoxin Biotechnology, Shanghai, China

  4. Jing Quan Dai

    Present address: Tayu Biotech, Beijing, China

  5. Qian Li

    Present address: Janssen (China) R&D Center, Shanghai, China

  6. Qian Gang Zheng

    Present address: EternBio, Shanghai, China

  7. Hui Xin Wan

    Present address: Ringene Pharmaceutical, Shanghai, China

  8. Feng He

    Present address: Hengrui Pharmaceutical, Shanghai, China

Authors and Affiliations

  1. Novartis Institutes for BioMedical Research, B4F2 Novartis Campus, No. 4218 Jinke Road, Shanghai, 201203, China

    Jiang Wei Zhang, Hai Bing Deng, Chun Ye Zhang, Jing Quan Dai, Qian Li, Qian Gang Zheng, Hui Xin Wan, Hong Ping Yu, Feng He, Yao Chang Xu, Sylvia Zhao & Ji Yue Jeff Zhang

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Correspondence to Jiang Wei Zhang or Ji Yue Jeff Zhang.

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All authors were or still are Novartis employee.

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All animal studies were in accordance with the ethical standards of the institution or practice at which the studies were conducted. All animal study procedures were approved by animal care and use committees before the studies were initiated.

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Zhang, J.W., Deng, H.B., Zhang, C.Y. et al. In Vitro Metabolism by Aldehyde Oxidase Leads to Poor Pharmacokinetic Profile in Rats for c-Met Inhibitor MET401. Eur J Drug Metab Pharmacokinet 44, 669–680 (2019). https://doi.org/10.1007/s13318-019-00557-9

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