Use of metabonomics to identify impaired fatty acid metabolism as the mechanism of a drug-induced toxicity.

An increased diversity of therapeutic targets in the pharmaceutical industry in recent years has led to a greater diversity of toxicological effects. This, and the increased pace of drug discovery, leads to a need for new technologies for the rapid elucidation of toxicological mechanisms. As part of an evaluation of the utility of metabonomics in drug safety assessment, 1H NMR spectra were acquired on urine and liver tissue samples obtained from rats administered vehicle or a development compound (MrkA) previously shown to induce hepatotoxicity in several animal species. Multivariate statistical analysis of the urinary NMR data clearly discriminated drug-treated from control animals, due to a depletion in tricarboxylic acid cycle intermediates, and the appearance of medium chain dicarboxylic acids. High-resolution magic angle spinning NMR data acquired on liver samples exhibited elevated triglyceride levels that were correlated with changes in the urinary NMR data. Urinary dicarboxylic aciduria is associated with defective metabolism of fatty acids; subsequent in vitro experiments confirmed that MrkA impairs fatty acid metabolism. The successful application of metabonomics to characterize an otherwise ill-defined mechanism of drug-induced toxicity supports the practicality of this approach for resolving toxicity issues for drugs in discovery and development.

[1]  E. De Clercq,et al.  Inhibition of HIV infection by CXCR4 and CCR5 chemokine receptor antagonists. , 2001, Antiviral chemistry & chemotherapy.

[2]  J. Nicholson,et al.  NMR and pattern recognition studies on liver extracts and intact livers from rats treated with alpha-naphthylisothiocyanate. , 2002, Biochemical pharmacology.

[3]  R. Chalmers,et al.  Medium-chain acyl-CoA dehydrogenase deficiency: a 1H-n.m.r. spectroscopic study. , 1990, Biochemical Society transactions.

[4]  P. Divry,et al.  General (medium-chain) acyl-CoA dehydrogenase deficiency (non-ketotic dicarboxylic aciduria): quantitative urinary excretion pattern of 23 biologically significant organic acids in three cases. , 1983, Clinica chimica acta; international journal of clinical chemistry.

[5]  K. Tanaka,et al.  Hypoglycin and Jamaican vomiting sickness. , 1990, Progress in clinical and biological research.

[6]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[7]  John C. Lindon,et al.  Detection of in vivo biomarkers of phospholipidosis using NMR‐based metabonomic approaches , 2001 .

[8]  D. Rabier,et al.  Recognition and management of fatty acid oxidation defects: A series of 107 patients , 1999, Journal of Inherited Metabolic Disease.

[9]  C. J. Van der Schyf,et al.  Metabolic defects caused by treatment with the tetrahydropyridine analog of haloperidol (HPTP), in baboons. , 1997, Life sciences.

[10]  E Holmes,et al.  Nuclear magnetic resonance spectroscopic and principal components analysis investigations into biochemical effects of three model hepatotoxins. , 1998, Chemical research in toxicology.

[11]  S. Kolvraa,et al.  On the Biologic Origin of C6-C10-Dicarboxylic and C6-C10-ω-1-Hydroxy Monocarboxylic Acids in Human and Rat with Acyl-CoA Dehydrogenation Deficiencies: in Vitro Studies on the ω- and ω-1-Oxidation of Medium-Chain (C6-C12) Fatty Acids in Human and Rat Liver , 1983, Pediatric Research.

[12]  A. J. Gandolfi,et al.  In vitro cytotoxicity of allyl alcohol and bromobenzene in a novel organ culture system. , 1987, Toxicology and applied pharmacology.

[13]  I. Wilson,et al.  Use of Proton NMR for Determining Changes in Metabolite Excretion Profiles Induced by Dietary Changes in the Rat , 1997 .

[14]  M. Carrington,et al.  Genetics of HIV-1 infection: chemokine receptor CCR5 polymorphism and its consequences. , 1999, Human molecular genetics.

[15]  G. Ellman.  TISSUE SULPHYDRYL GROUPS , 1959 .

[16]  Richard S Paules,et al.  An overview of toxicogenomics. , 2002, Current issues in molecular biology.

[17]  J. Lindon,et al.  Metabonomics: a platform for studying drug toxicity and gene function , 2002, Nature Reviews Drug Discovery.

[18]  N. Maeda,et al.  Defects in the Generation of IFN-γ Are Overcome to Control Infection with Leishmania donovani in CC Chemokine Receptor (CCR) 5-, Macrophage Inflammatory Protein-1α-, or CCR2-Deficient Mice , 1999, The Journal of Immunology.

[19]  E Holmes,et al.  High-resolution magic angle spinning (1)H NMR spectroscopy of intact liver and kidney: optimization of sample preparation procedures and biochemical stability of tissue during spectral acquisition. , 2000, Analytical biochemistry.

[20]  J. Nicholson,et al.  High‐resolution 1H and 1H‐13C magic angle spinning NMR spectroscopy of rat liver , 2000, Magnetic resonance in medicine.

[21]  M. Hammer,et al.  Global survey of genetic variation in CCR5, RANTES, and MIP-1α: Impact on the epidemiology of the HIV-1 pandemic , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[22]  E Holmes,et al.  NMR and pattern recognition studies on the time-related metabolic effects of alpha-naphthylisothiocyanate on liver, urine, and plasma in the rat: an integrative metabonomic approach. , 2001, Chemical research in toxicology.

[23]  E Holmes,et al.  High‐resolution magic angle spinning 1H NMR spectroscopic studies on intact rat renal cortex and medulla , 1999, Magnetic resonance in medicine.

[24]  Elaine Holmes,et al.  Metabonomic applications in toxicity screening and disease diagnosis. , 2002, Current topics in medicinal chemistry.

[25]  J. Nicholson,et al.  A metabonomic approach to the investigation of drug-induced phospholipidosis: an NMR spectroscopy and pattern recognition study , 2000 .

[26]  S. Wold,et al.  Multi‐way principal components‐and PLS‐analysis , 1987 .

[27]  J. Farber,et al.  HIV's response to a CCR5 inhibitor: I'd rather tighten than switch! , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[28]  L. Cucchiarini,et al.  A very fast ion-pair reversed-phase HPLC method for the separation of the most significant nucleotides and their degradation products in human red blood cells. , 1987, Analytical biochemistry.

[29]  I. Wilson,et al.  Effect of diet on the urinary excretion of hippuric acid and other dietary-derived aromatics in rat. A complex interaction between diet, gut microflora and substrate specificity. , 1998, Xenobiotica; the fate of foreign compounds in biological systems.

[30]  Thomas Linke,et al.  Visualizing plant metabolomic correlation networks using clique-metabolite matrices , 2001, Bioinform..

[31]  H. Kröger,et al.  Methylation of nicotinamide in rat liver cytosol and its correlation with hepatocellular proliferation. , 1982, Biochimica et biophysica acta.

[32]  S. Connor,et al.  Antidiabetic Efficacy of BRL 49653, a Potent Orally Active Insulin Sensitizing Agent, Assessed in the C57BL/KsJ db/db Diabetic Mouse by Non‐invasive 1H NMR Studies of Urine , 1997, The Journal of pharmacy and pharmacology.

[33]  M. Reily,et al.  Effect of dexamethasone on the metabonomics profile associated with phosphodiesterase inhibitor-induced vascular lesions in rats. , 2002, Toxicology and applied pharmacology.

[34]  C. I. Pogson,et al.  The flux control coefficient of carnitine palmitoyltransferase I on palmitate beta-oxidation in rat hepatocyte cultures. , 1997, The Biochemical journal.

[35]  R. Mortishire-Smith,et al.  Temperature calibration of a high‐resolution magic‐angle spinning NMR probe for analysis of tissue samples , 2001 .

[36]  J. Lindon,et al.  The identification of novel biomarkers of renal toxicity using automatic data reduction techniques and PCA of proton NMR spectra of urine , 1998 .