Sensitivity of (1)H NMR analysis of rat urine in relation to toxicometabonomics. Part I: dose-dependent toxic effects of bromobenzene and paracetamol.

(1)H nuclear magnetic resonance (NMR) spectroscopy of rat urine in combination with pattern recognition analysis was evaluated for early noninvasive detection of toxicity of investigational chemical entities. Bromobenzene (B) and paracetamol (P) were administered at five single oral dosages between 2 and 500 mg/kg and between 6 and 1800 mg/kg, respectively. The sensitivity of the proposed method to detect changes in the NMR spectra 24 and 48 h after single dosing was compared with histopathology and biochemical parameters in plasma and urine. Both B and P applied at the highest dosages induced liver necrosis and markedly increased aspartate aminotransferase (AST) and alanine aminotransferase (ALT) plasma levels. At dosages of 125 mg/kg B and 450 mg/kg P, liver necrosis and changes in AST and ALT were less pronounced, while at lower dose levels these effects could not be detected. Changes in kidney pathology or standard urine biochemistry were not observed at any of these dosages. Evaluation of the total NMR dataset showed 80 signals to be sensitive for B and P dosing. Principal component analysis on the reduced dataset revealed that NMR spectra were significantly different at dosages above 8 mg/kg (B) and 110 mg/kg (P) at both sampling times. This implies a 4- to 16-fold increased sensitivity of NMR versus histopathology and clinical chemistry in recognizing early events of liver toxicity.

[1]  Procrustes rotation and pair-wise correlation: A parametric and a non-parametric method for variable selection , 2004 .

[2]  J. van der Greef,et al.  Evaluation of metabolite profiles as biomarkers for the pharmacological effects of thiazolidinediones in Type 2 diabetes mellitus patients and healthy volunteers. , 2007, British journal of clinical pharmacology.

[3]  D. Jollow,et al.  Bromobenzene-induced liver necrosis. Protective role of glutathione and evidence for 3,4-bromobenzene oxide as the hepatotoxic metabolite. , 1974, Pharmacology.

[4]  C. Rae,et al.  1H NMR spectroscopic survey of plasma and erythrocytes from selected marsupials and domestic animals of Australia. , 1991, Comparative biochemistry and physiology. B, Comparative biochemistry.

[5]  A. Wittwer,et al.  Identification of folate binding protein of mitochondria as dimethylglycine dehydrogenase. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[6]  E. Will,et al.  Use of the urinary protein creatinine index to assess proteinuria in renal transplant patients. , 1992, Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association.

[7]  W. Windig,et al.  Interpretation of sets of pyrolysis mass spectra by discriminant analysis and graphical rotation , 1983 .

[8]  K. Siamopoulos,et al.  Paraquat-induced renal injury studied by 1H nuclear magnetic resonance spectroscopy of urine. , 1998, Clinical chemistry.

[9]  J. van der Greef,et al.  Partial linear fit: A new NMR spectroscopy preprocessing tool for pattern recognition applications , 1996 .

[10]  C P Siegers,et al.  Relations between hepatotoxicity and pharmacokinetics of paracetamol in rats and mice. , 1978, Pharmacology.

[11]  Esther F. Schmid,et al.  Drug withdrawals and the lessons within. , 2006, Current opinion in drug discovery & development.

[12]  M. Reily,et al.  Metabonomics: evaluation of nuclear magnetic resonance (NMR) and pattern recognition technology for rapid in vivo screening of liver and kidney toxicants. , 2000, Toxicological sciences : an official journal of the Society of Toxicology.

[13]  Jimmy D Bell,et al.  Urinary levels of creatine and other metabolites in the assessment of polymyositis and dermatomyositis. , 2003, Rheumatology.

[14]  J. Lindon,et al.  Pharmaco-metabonomic phenotyping and personalized drug treatment , 2006, Nature.

[15]  P. G. Kistemaker,et al.  Discriminant analysis by double stage principal component analysis , 1983 .

[16]  S. Narayanan,et al.  Creatinine: a review. , 1980, Clinical chemistry.

[17]  J C Lindon,et al.  Application of pattern recognition methods to the analysis and classification of toxicological data derived from proton nuclear magnetic resonance spectroscopy of urine. , 1991, Molecular pharmacology.

[18]  J. Griffin,et al.  Metabolic profiling of chronic cadmium exposure in the rat. , 2001, Chemical research in toxicology.

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

[20]  D. Ritchie,et al.  The new pre-preclinical paradigm: compound optimization in early and late phase drug discovery. , 2001, Current topics in medicinal chemistry.

[21]  John C. Lindon,et al.  Hepatotoxin-induced hypercreatinaemia and hypercreatinuria: their relationship to one another, to liver damage and to weakened nutritional status , 2004, Archives of Toxicology.

[22]  C. Laggner,et al.  Why drugs fail--a study on side effects in new chemical entities. , 2005 .

[23]  J. Lindon,et al.  An hypothesis for a mechanism underlying hepatotoxin-induced hypercreatinuria , 2003, Archives of Toxicology.

[24]  T. Williams,et al.  Identification of three protein targets for reactive metabolites of bromobenzene in rat liver cytosol. , 2000, Chemical research in toxicology.

[25]  T. Fan Metabolite profiling by one- and two-dimensional NMR analysis of complex mixtures , 1996 .

[26]  John C. Lindon,et al.  NMR-based metabonomic studies on the biochemical effects of commonly used drug carrier vehicles in the rat. , 2002 .

[27]  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.

[28]  D. Massart Chemometrics: A Textbook , 1988 .

[29]  J. T. W. E. Vogels A new method for classification of wines based on proton and carbon-13 NMR spectroscopy in combination with pattern recognition techniques. Chemometrics and Intelligent Laboratory Systems , 1993 .

[30]  Ben van Ommen,et al.  Profiles of Metabolites and Gene Expression in Rats with Chemically Induced Hepatic Necrosis , 2005, Toxicologic pathology.

[31]  Maurice M. Tatsuoka,et al.  Multivariate Statistics for Personnel Classification. , 1968 .

[32]  Matej Orešič,et al.  The Role of Metabolomics in Systems Biology , 2003 .

[33]  V. Govindaraju,et al.  Proton NMR chemical shifts and coupling constants for brain metabolites , 2000, NMR in biomedicine.

[34]  B. E. Walker,et al.  Serum transaminase levels after experimental paracetamol-induced hepatic necrosis. , 1975, Gut.

[35]  J. Trygg,et al.  Evaluation of the orthogonal projection on latent structure model limitations caused by chemical shift variability and improved visualization of biomarker changes in 1H NMR spectroscopic metabonomic studies. , 2005, Analytical chemistry.

[36]  Jeremy K. Nicholson,et al.  Biochemical characterisation of para-aminophenol-induced nephrotoxic lesions in the F344 rat , 2004, Archives of Toxicology.

[37]  J. Lindon,et al.  NMR‐based metabonomic approaches for evaluating physiological influences on biofluid composition , 2005, NMR in biomedicine.

[38]  R. Billings,et al.  Bromobenzene metabolism in vivo and in vitro. The mechanism of 4-bromocatechol formation. , 1990, Drug metabolism and disposition: the biological fate of chemicals.

[39]  J. Lindon,et al.  'Metabonomics': understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data. , 1999, Xenobiotica; the fate of foreign compounds in biological systems.

[40]  J R Gillette,et al.  Role of detoxifying enzymes in bromobenzene-induced liver necrosis. , 1973, The Journal of pharmacology and experimental therapeutics.

[41]  Bruce R. Kowalski,et al.  Measurement analysis by pattern recognition , 1975 .

[42]  A. Wittwer,et al.  Identification of the folate-binding proteins of rat liver mitochondria as dimethylglycine dehydrogenase and sarcosine dehydrogenase. Flavoprotein nature and enzymatic properties of the purified proteins. , 1981, The Journal of biological chemistry.

[43]  J. Joly,et al.  Effect of chronic ethanol administration on bromobenzene liver toxicity in the rat. , 1983, Toxicology and applied pharmacology.

[44]  A. Tas,et al.  Uniform procedure of (1)H NMR analysis of rat urine and toxicometabonomics Part II: comparison of NMR profiles for classification of hepatotoxicity. , 2007, Toxicological sciences : an official journal of the Society of Toxicology.

[45]  E Holmes,et al.  Chemometric models for toxicity classification based on NMR spectra of biofluids. , 2000, Chemical research in toxicology.

[46]  R. Goodacre,et al.  Metabolic Profiling: Its Role in Biomarker Discovery and Gene Function Analysis , 2003, Springer US.

[47]  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.

[48]  Pedro Cuatrecasas,et al.  Drug discovery in jeopardy. , 2006, The Journal of clinical investigation.

[49]  J. Tredger,et al.  Effects of ethanol ingestion on the hepatotoxicity and metabolism of paracetamol in mice. , 1985, Toxicology.

[50]  W. Hwu,et al.  Cloning of dimethylglycine dehydrogenase and a new human inborn error of metabolism, dimethylglycine dehydrogenase deficiency. , 2001, American journal of human genetics.

[51]  R. Niesink,et al.  Toxicology: Principles and Applications , 1996 .