RESUMO
A major purpose of exploratory metabolic profiling is for the identification of molecular species that are statistically associated with specific biological or medical outcomes; unfortunately, the structure elucidation process of unknowns is often a major bottleneck in this process. We present here new holistic strategies that combine different statistical spectroscopic and analytical techniques to improve and simplify the process of metabolite identification. We exemplify these strategies using study data collected as part of a dietary intervention to improve health and which elicits a relatively subtle suite of changes from complex molecular profiles. We identify three new dietary biomarkers related to the consumption of peas (N-methyl nicotinic acid), apples (rhamnitol), and onions (N-acetyl-S-(1Z)-propenyl-cysteine-sulfoxide) that can be used to enhance dietary assessment and assess adherence to diet. As part of the strategy, we introduce a new probabilistic statistical spectroscopy tool, RED-STORM (Resolution EnhanceD SubseT Optimization by Reference Matching), that uses 2D J-resolved 1H NMR spectra for enhanced information recovery using the Bayesian paradigm to extract a subset of spectra with similar spectral signatures to a reference. RED-STORM provided new information for subsequent experiments (e.g., 2D-NMR spectroscopy, solid-phase extraction, liquid chromatography prefaced mass spectrometry) used to ultimately identify an unknown compound. In summary, we illustrate the benefit of acquiring J-resolved experiments alongside conventional 1D 1H NMR as part of routine metabolic profiling in large data sets and show that application of complementary statistical and analytical techniques for the identification of unknown metabolites can be used to save valuable time and resources.
Assuntos
Malus/metabolismo , Ácidos Nicotínicos/análise , Cebolas/metabolismo , Pisum sativum/metabolismo , Ramnose/análise , Biomarcadores/análise , Biomarcadores/metabolismo , Espectroscopia de Ressonância Magnética , Malus/química , Estrutura Molecular , Ácidos Nicotínicos/metabolismo , Cebolas/química , Pisum sativum/química , Ramnose/análogos & derivados , Ramnose/metabolismoRESUMO
Most chronic diseases impairing current human public health involve not only the human genome but also gene-environment interactions, and in the latter case the gut microbiome is an important factor. This makes the classical single drug-receptor target drug discovery paradigm much less applicable. There is widespread and increasing international interest in understanding the properties of traditional Chinese medicines (TCMs) for their potential utilization as a source of new drugs for Western markets as emerging evidence indicates that most TCM drugs are actually targeting both the host and its symbiotic microbes. In this review, we explore the challenges of and opportunities for harmonizing Eastern-Western drug discovery paradigms by focusing on emergent functions at the whole body level of humans as superorganisms. This could lead to new drug candidate compounds for chronic diseases targeting receptors outside the currently accepted "druggable genome" and shed light on current high interest issues in Western medicine such as drug-drug and drug-diet-gut microbial interactions that will be crucial in the development and delivery of future therapeutic regimes optimized for the individual patient.
Assuntos
Descoberta de Drogas , Genoma Humano , Medicina Tradicional Chinesa , Metagenoma , Animais , Doença Crônica/tratamento farmacológico , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/uso terapêutico , Trato Gastrointestinal/microbiologia , Humanos , Metagenoma/efeitos dos fármacos , Biologia de SistemasRESUMO
In traditional Chinese medicine (TCM), multicomponent and principally plant-derived drugs are used for disease prevention, symptom amelioration and treatment in a personalized manner. Because of their complex composition and consequent multiple targets and treatment objectives, the application of omics techniques and other integrative approaches seems inherently appropriate and even necessary for the demonstration of their potential preclinical and clinical safety and efficacy. This perspectives article provides proposals for the application of omics methods to the investigation of complex herbal products (CHP),(1) including Chinese herbal medicines (CHM), both in vitro and in vivo, for preclinical and clinical toxicity, pharmacokinetics, pharmacodynamics and efficacy tests. Ultimately, such approaches could aid regulatory scrutiny and potential acceptance, although currently there is no regulatory requirement of omics-based data in any submitted dossier to any regulatory agency, including for conventional drugs and CHP. However, it has been acknowledged that such studies are being increasingly performed, and almost surely will eventually be included into regulatory submission dossiers, possibly initially as supplementary materials. Specifically for CHM and CHP, omics can play a role both in determining product composition and its variability and in monitoring biological effects in carefully selected platforms. Predicting the future is difficult, but it seems possible that regulatory acceptance of omics techniques and a systems biology approach for the study of TCM, CHM and CHP will not be long delayed. It is expected that current studies and plans employing omics techniques and other integrative approaches will prove to be positive and informative.
Assuntos
Pesquisa Biomédica/métodos , Medicamentos de Ervas Chinesas , Regulamentação Governamental , Medicina Herbária/métodos , Legislação de Medicamentos , Fitoterapia , Tecnologia Farmacêutica/métodos , Animais , Qualidade de Produtos para o Consumidor , Medicamentos de Ervas Chinesas/efeitos adversos , Medicamentos de Ervas Chinesas/uso terapêutico , Medicina Herbária/legislação & jurisprudência , Humanos , Medicina Tradicional Chinesa , Plantas Medicinais , Projetos de Pesquisa , Tecnologia Farmacêutica/legislação & jurisprudênciaRESUMO
Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) provides localized information about the molecular content of a tissue sample. To derive reliable conclusions from MSI data, it is necessary to implement appropriate processing steps in order to compare peak intensities across the different pixels comprising the image. Here, we review commonly used normalization methods, and propose a rational data processing strategy, for robust evaluation and modeling of MSI data. The approach includes newly developed heuristic methods for selecting biologically relevant peaks and pixels to reduce the size of a data set and remove the influence of the applied MALDI matrix. The methods are demonstrated on a MALDI MSI data set of a sagittal section of rat brain (4750 bins, m/z = 50-1000, 111 × 185 pixels) and the proposed preferred normalization method uses the median intensity of selected peaks, which were determined to be independent of the MALDI matrix. This was found to effectively compensate for a range of known limitations associated with the MALDI process and irregularities in MS image sampling routines. This new approach is relevant for processing of all MALDI MSI data sets, and thus likely to have impact in biomarker profiling, preclinical drug distribution studies, and studies addressing underlying molecular mechanisms of tissue pathology.
Assuntos
Processamento de Imagem Assistida por Computador/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Animais , Biomarcadores/metabolismo , Encéfalo/patologia , Avaliação Pré-Clínica de Medicamentos , Processamento de Imagem Assistida por Computador/normas , Análise de Componente Principal , RatosRESUMO
The effects of sample preparation and chromatographic method differences on the classification and recovery of metabolic biomarkers from UPLC-MS measurements on urine samples of humans exposed to different dietary interventions have been investigated. Eight volunteers consumed three high-fat meals (rich in saturated, monounsaturated and polyunsaturated fatty acids, respectively) in randomized order with a washout period in between. For each participant, urine samples were obtained prior to and at three timed intervals after each meal. Samples were processed either by dilution (1 : 4) or by liquid-liquid extraction and then run under two different gradient conditions. For each analysis method, a total of 96 observations (eight participants, four time points, three diets) were measured. The total ion count chromatograms were analyzed using partial-least-squares discriminant analysis. All three dietary classes could be discriminated irrespective of sample preparation and chromatographic method. However, the main discriminating metabolites varied according to sample preparation, indicating that sample treatment and chromatographic conditions influence the ability to extract biomolecular information. Diluted samples showed higher m/z compounds (ca 400 u) while liquid-liquid extraction samples showed low m/z at the same retention time span. Optimized methods for metabolite identification (e.g. organic acids) were statistically inferior to global screening for mixed compound identification, confirming that multiple compound class-based metabolic profiles are likely to give superior metabonomic (diagnostic) classification, although great care has to be taken in the interpretation in relation to matrix effects.
Assuntos
Cromatografia Líquida/métodos , Gorduras na Dieta , Ácidos Graxos Monoinsaturados/urina , Ácidos Graxos Insaturados/urina , Ácidos Graxos/urina , Espectrometria de Massas/métodos , Metabolômica , Adolescente , Humanos , Masculino , Período Pós-Prandial , Adulto JovemRESUMO
The first application of high field NMR spectroscopy (800 MHz for (1)H observation) to human hepatic bile (as opposed to gall bladder bile) is reported. The bile sample used for detailed investigation was from a donor liver with mild fat infiltration, collected during organ retrieval prior to transplantation. In addition, to focus on the detection of bile acids in particular, a bile extract was analysed by 800 MHz (1)H NMR spectroscopy, HPLC-NMR/MS and UPLC-MS. In the whole bile sample, 40 compounds have been assigned with the aid of two-dimensional (1)H-(1)H TOCSY and (1)H-(13)C HSQC spectra. These include phosphatidylcholine, 14 amino acids, 10 organic acids, 4 carbohydrates and polyols (glucose, glucuronate, glycerol and myo-inositol), choline, phosphocholine, betaine, trimethylamine-N-oxide and other small molecules. An initial NMR-based assessment of the concentration range of some key metabolites has been made. Some observed chemical shifts differ from expected database values, probably due to a difference in bulk diamagnetic susceptibility. The NMR spectra of the whole extract gave identification of the major bile acids (cholic, deoxycholic and chenodeoxycholic), but the glycine and taurine conjugates of a given bile acid could not be distinguished. However, this was achieved by HPLC-NMR/MS, which enabled the separation and identification of ten conjugated bile acids with relative abundances varying from approximately 0.1% (taurolithocholic acid) to 34.0% (glycocholic acid), of which, only the five most abundant acids could be detected by NMR, including the isomers glycodeoxycholic acid and glycochenodeoxycholic acid, which are difficult to distinguish by conventional LC-MS analysis. In a separate experiment, the use of UPLC-MS allowed the detection and identification of 13 bile acids. This work has shown the complementary potential of NMR spectroscopy, MS and hyphenated NMR/MS for elucidating the complex metabolic profile of human hepatic bile. This will be useful baseline information in ongoing studies of liver excretory function and organ transplantation.
Assuntos
Ácidos e Sais Biliares/química , Bile/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Fígado/metabolismo , Espectroscopia de Ressonância Magnética/métodos , Espectrometria de Massas/métodos , Cromatografia Líquida/métodos , Fígado Gorduroso/metabolismo , Humanos , Metaboloma , Modelos Químicos , Reprodutibilidade dos TestesRESUMO
The time-related metabolic responses to l-arginine (ARG)-induced exocrine pancreatic toxicity were investigated using single ip doses of 1,000 and 4,000 mg/kg body weight over a 7 day experimental period in male Sprague-Dawley rats. Sequential timed urine and plasma samples were analyzed using high resolution (1)H NMR spectroscopy together with complementary clinical chemistry and histopathology analyses. Principal components analysis (PCA) and orthogonal projection on latent structures discriminant analysis (O-PLS-DA) were utilized to analyze the (1)H NMR data and to extract and identify candidate biomarkers and to construct metabolic trajectories post ARG administration. Low doses of ARG resulted in virtually no histopathological damage and distinct reversible metabolic response trajectories. High doses of ARG caused pancreatic acinar degeneration and necrosis and characteristic metabolic trajectory profiles with several distinct phases. The initial trajectory phase (0-8 h) involved changes in the urea cycle and transamination indicating a homeostatic response to detoxify excess ammonia generated from ARG catabolism. By 48 h, there was a notable enhancement of the excretion of the gut microbial metabolites, phenylacetylglycine (PAG), 4-cresol-glucuronide and 4-cresol-sulfate, suggesting that compromised pancreatic function impacts on the activity of the gut microbiota giving potential rise to a novel class of surrogate extragenomic biomarkers of pancreatic injury. The implied compromise of microbiotal function may also contribute to secondary hepatic and pancreatic toxic responses. We show here for the first time the value of metabonomic studies in investigating metabolic disruption due to experimental pancreatitis. The variety of observed systemic responses suggests that this approach may be of general value in the assessment of other animal models or human pancreatitis.
Assuntos
Arginina/toxicidade , Metabolismo , Modelos Biológicos , Pancreatite/induzido quimicamente , Animais , Biomarcadores/sangue , Biomarcadores/urina , Humanos , Fígado/metabolismo , Fígado/patologia , Masculino , Ressonância Magnética Nuclear Biomolecular , Pancreatite/metabolismo , Pancreatite/patologia , Distribuição Aleatória , Ratos , Ratos Sprague-DawleyRESUMO
Gut microbiome-host metabolic interactions affect human health and can be modified by probiotic and prebiotic supplementation. Here, we have assessed the effects of consumption of a combination of probiotics (Lactobacillus paracasei or L. rhamnosus) and two galactosyl-oligosaccharide prebiotics on the symbiotic microbiome-mammalian supersystem using integrative metabolic profiling and modeling of multiple compartments in germ-free mice inoculated with a model of human baby microbiota. We have shown specific impacts of two prebiotics on the microbial populations of HBM mice when co-administered with two probiotics. We observed an increase in the populations of Bifidobacterium longum and B. breve, and a reduction in Clostridium perfringens, which were more marked when combining prebiotics with L. rhamnosus. In turn, these microbial effects were associated with modulation of a range of host metabolic pathways observed via changes in lipid profiles, gluconeogenesis, and amino-acid and methylamine metabolism associated to fermentation of carbohydrates by different bacterial strains. These results provide evidence for the potential use of prebiotics for beneficially modifying the gut microbial balance as well as host energy and lipid homeostasis.
Assuntos
Genoma/genética , Intestinos/microbiologia , Lactobacillus/genética , Lactobacillus/metabolismo , Modelos Animais , Probióticos , Biologia de Sistemas , Animais , Peso Corporal , Ceco/metabolismo , Ácidos Graxos/metabolismo , Fezes/microbiologia , Feminino , Genoma/efeitos dos fármacos , Humanos , Lactente , Intestinos/efeitos dos fármacos , Fígado/metabolismo , Espectroscopia de Ressonância Magnética , Camundongos , Probióticos/farmacologiaRESUMO
Detection and classification of in vivo drug toxicity is an expensive and time-consuming process. Metabolic profiling is becoming a key enabling tool in this area as it provides a unique perspective on the characterization and mechanisms of response to toxic insult. As part of the Consortium on Metabonomic Toxicology (COMET) project, a substantial metabolic and pathological database was constructed. We chose a set of 80 treatments to build a modeling system for toxicity prediction using NMR spectroscopy of urine samples (n=12935) from laboratory rats (n=1652). The compound structures and activities were diverse but there was an emphasis on the selection of hepato and nephrotoxins. We developed a two-stage strategy based on the assumptions that (a) adverse effects would produce metabolic profiles deviating from those of normal animals and (b) such deviations would be similar for treatments having similar physiological effects. To address the first stage, we developed a multivariate model of normal urine, using principal components analysis of specially preprocessed 1H NMR spectra. The model demonstrated a high correspondence between the occurrence of toxicity and abnormal metabolic profiles. In the second stage, we extended a density estimation method, "CLOUDS", to compute multidimensional similarities between treatments. Crucially, the technique allowed a distribution-free estimate of similarity across multiple animals and time points for each treatment and the resulting matrix of similarities showed segregation between liver toxins and other treatments. Using the similarity matrix, we were able to correctly identify the target organ of two "blind" treatments, even at sub-toxic levels. To further validate the approach, we then applied a leave-one-out approach to predict the main organ of toxicity (liver or kidney) showing significant responses using the three most similar matches in the matrix. Where predictions could be made, there was an error rate of 8%. The sensitivities to liver and kidney toxicity were 67 and 41%, respectively, whereas the corresponding specificities were 77 and 100%. In some cases, it was not possible to make predictions because of interference by drug-related metabolite signals (18%), an inconsistent histopathological or urinary response (11%), genuine class overlap (8%), or lack of similarity to any other treatment (2%). This study constitutes the largest validation to date of the metabonomic approach to preclinical toxicology assessment, confirming that the methodology offers practical utility for rapid in vivo drug toxicity screening.
Assuntos
Química Farmacêutica/métodos , Avaliação Pré-Clínica de Medicamentos/instrumentação , Avaliação Pré-Clínica de Medicamentos/métodos , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Tecnologia Farmacêutica/métodos , Toxicologia/métodos , Animais , Humanos , Rim/efeitos dos fármacos , Fígado/efeitos dos fármacos , Espectroscopia de Ressonância Magnética/métodos , Masculino , Modelos Estatísticos , Probabilidade , Ratos , Ratos Sprague-Dawley , Sensibilidade e Especificidade , Fatores de TempoRESUMO
We have used a simplified gnotobiotic mouse model to evaluate the effects of single bacterial species, Lactobacillus paracasei NCC2461, on the metabolic profiles of intact intestinal tissues using high-resolution magic-angle-spinning 1H NMR spectroscopy (HRMAS). A total of 24 female gnotobiotic mice were divided into three groups: a control group supplemented with water and two groups supplemented with either live L. paracasei or a gamma-irradiated equivalent. HRMAS was used to characterize the biochemical components of intact epithelial tissues from the duodenum, jejunum, ileum, proximal, and distal colons in all animals and data were analyzed using chemometrics. Variations in relative concentrations of amino acids, anti-oxidant, and creatine were observed relating to different physiological properties in each intestinal tissue. Metabolic characteristics of lipogenesis and fat storage were observed in the jejunum and colon. Colonization with live L. paracasei induced region-dependent changes in the metabolic profiles of all intestinal tissues, except for the colon, consistent with modulation of intestinal digestion, absorption of nutrients, energy metabolism, lipid synthesis and protective functions. Ingestion of gamma-irradiated bacteria produced no effects on the observed metabolic profiles. 1H MAS NMR spectroscopy was able to generate characteristic metabolic signatures reflecting the structure and function of intestinal tissues. These signals acted as reference profiles with which to compare changes in response to gut microbiota manipulation at the tissue level as demonstrated by ingestion of a bacterial probiotic.
Assuntos
Mucosa Intestinal/metabolismo , Intestinos/efeitos dos fármacos , Lactobacillus , Ressonância Magnética Nuclear Biomolecular/métodos , Probióticos/farmacologia , Aminoácidos/análise , Aminoácidos/metabolismo , Animais , Metabolismo Energético , Feminino , Vida Livre de Germes , Inflamação/metabolismo , Intestinos/microbiologia , Lactobacillus/efeitos da radiação , Lipídeos/análise , Lipídeos/biossíntese , Camundongos , Camundongos Endogâmicos , Contração Muscular , Probióticos/efeitos da radiaçãoRESUMO
This minireview is based on a lecture given at the First Maga Circe Conference on metabolomics held at Sabaudia, Italy, in March 2006 in which the analytical and statistical techniques used in metabonomics, efforts at standardization and some of the major applications to pharmaceutical research and development are reviewed. Metabonomics involves the determination of multiple metabolites simultaneously in biofluids, tissues and tissue extracts. Applications to preclinical drug safety studies are illustrated by the Consortium for Metabonomic Toxicology, a collaboration involving several major pharmaceutical companies. This consortium was able, through the measurement of a dataset of NMR spectra of rodent urine and serum samples, to build a predictive expert system for liver and kidney toxicity. A secondary benefit was the elucidation of the endogenous biochemicals responsible for the classification. The use of metabonomics in disease diagnosis and therapy monitoring is discussed with an exemplification from coronary artery disease, and the concept of pharmaco-metabonomics as a way of predicting an individual's response to treatment is exemplified. Finally, some advantages and perceived difficulties of the metabonomics approach are summarized.
Assuntos
Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Tecnologia Farmacêutica/métodos , Animais , Biomarcadores/análise , Avaliação Pré-Clínica de Medicamentos , Previsões , Humanos , Espectroscopia de Ressonância Magnética/métodos , Espectrometria de Massas/métodos , Preparações Farmacêuticas/química , Preparações Farmacêuticas/metabolismo , Farmacogenética , FarmacologiaRESUMO
Irritable Bowel Syndrome (IBS) is a common multifactorial intestinal disorder for which the aetiology remains largely undefined. Here, we have used a Trichinella spiralis (T. spiralis)-induced model of post-infective IBS, and the effects of probiotic bacteria on gut dysfunction have been investigated using a metabonomic strategy. A total of 44 mice were divided into four groups: an uninfected control group and three T. spiralis-infected groups, one as infected control and the two other groups subsequently treated with either Lactobacillus paracasei (L. paracasei) NCC2461 in spent culture medium (SCM) or with L. paracasei-free SCM. Plasma, jejunal wall and longitudinal myenteric muscle samples were collected at day 21 post-infection. An NMR-based metabonomic approach characterized that the plasma metabolic profile of T. spiralis-infected mice showed an increased energy metabolism (lactate, citrate, alanine), fat mobilization (acetoacetate, 3-D-hydroxybutyrate, lipoproteins) and a disruption of amino acid metabolism due to increased protein breakdown, which were related to the intestinal hypercontractility. Increased levels of taurine, creatine and glycerophosphorylcholine in the jejunal muscles were associated with the muscular hypertrophy and disrupted jejunal functions. L. paracasei treatment normalized the muscular activity and the disturbed energy metabolism as evidenced by decreased glycogenesis and elevated lipid breakdown in comparison with untreated T. spiralis-infected mice. Changes in the levels of plasma metabolites (glutamine, lysine, methionine) that might relate to a modulation of immunological responses were also observed in the presence of the probiotic treatment. The work presented here suggests that probiotics may be beneficial in patients with IBS.
Assuntos
Síndrome do Intestino Irritável/terapia , Lactobacillus/metabolismo , Probióticos/uso terapêutico , Trichinella spiralis , Triquinelose/metabolismo , Aminoácidos/sangue , Animais , Proteínas Sanguíneas/análise , Metabolismo Energético/fisiologia , Camundongos , Ressonância Magnética Nuclear Biomolecular , Triquinelose/sangueRESUMO
Stress in the form of moderate periods of maternal separation of newborn rats has been postulated to cause permanent changes in the central nervous system and diseases in later life. It is also considered that dietary supplementation with long chain polyunsaturated fatty acids (LC-PUFAs) can potentially ameliorate the effects of stress. The metabolic consequences of early life maternal separation stress were investigated in rats (2-14 days after birth), either alone or in combination with secondary acute water avoidance stress at 3-4 months of age. The effect of a LC-PUFA-enriched dietary intervention in stressed animals was also assessed. Systematic changes in metabolic biochemistry were evaluated using 1H nuclear magnetic resonance spectroscopy of blood plasma and multivariate pattern recognition techniques. The biochemical response to stress was characterized by decreased levels of total lipoproteins and increased levels of amino acids, glucose, lactate, creatine, and citrate. Secondary acute water avoidance stress also caused elevated levels of O-acetyl glycoproteins in blood plasma. LC-PUFAs dietary enrichment did not alter the metabolic response to stress, but did result in a modified lipoprotein profile. This work indicates that the different stressor types resulted in some common systemic metabolic responses that involve changes in energy and muscle metabolism, but that they are not reversible by dietary intervention.
Assuntos
Aprendizagem da Esquiva , Dieta , Gorduras Insaturadas na Dieta/administração & dosagem , Ácidos Graxos Insaturados/metabolismo , Estresse Psicológico , Aminoácidos/sangue , Animais , Animais Recém-Nascidos , Glicemia/análise , Ácido Cítrico/sangue , Creatinina/sangue , Ácidos Graxos Insaturados/sangue , Análise de Fourier , Glicoproteínas/sangue , Ácido Láctico/sangue , Lipoproteínas/sangue , Masculino , Modelos Biológicos , Ressonância Magnética Nuclear Biomolecular , Ratos , Ratos Long-EvansRESUMO
In this review, the background to the approach known as metabonomics is provided, giving a brief historical perspective and summarizing the analytical and statistical techniques used. Some of the major applications of metabonomics relevant to pharmaceutical Research & Development are then reviewed including the study of various influences on metabolism, such as diet, lifestyle, and other environmental factors. The applications of metabonomics in drug safety studies are explained with special reference to the aims and achievements of the Consortium for Metabonomic Toxicology. Next, the role that metabonomics might have in disease diagnosis and therapy monitoring is provided with some examples, and the concept of pharmacometabonomics as a way of predicting an individual's response to treatment is highlighted. Some discussion is given on the strengths and weaknesses, opportunities of, and threats to metabonomics.
Assuntos
Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Farmacocinética , Farmacologia , Biologia de Sistemas/métodos , Tecnologia Farmacêutica/métodos , Algoritmos , Animais , Biomarcadores/análise , Biotransformação , Líquidos Corporais/química , Líquidos Corporais/metabolismo , Avaliação Pré-Clínica de Medicamentos , Humanos , Espectroscopia de Ressonância Magnética/métodos , Espectrometria de Massas/métodos , Preparações Farmacêuticas/química , Preparações Farmacêuticas/metabolismo , Análise de Componente Principal , ProteômicaRESUMO
The utility of metabonomics in the evaluation of xenobiotic toxicity has been comprehensively assessed by the Consortium for Metabonomic Toxicology (COMET), formed between five major pharmaceutical companies and Imperial College London, UK. The main objectives were to assess methodologies, to generate a metabonomic database using (1)H nuclear magnetic resonance (NMR) spectroscopy of rodent urine and blood serum and to build a predictive expert system for target organ toxicity. The analytic and biologic variation that might arise through the use of metabonomics was evaluated and a high degree of robustness demonstrated. With the completion of 147 studies, the chief deliverables of a curated database of rodent biofluid NMR spectra and computer-based expert systems for the prediction of kidney or liver toxicity in rat and mouse based on the spectral data have been generated, and delivered to the sponsoring companies. The project, with its relatively modest resources, has met and exceeded all of its targets and was judged a resounding success by the sponsoring companies who are, in many cases, already enhancing and making use of the data in their in-house studies.
Assuntos
Avaliação Pré-Clínica de Medicamentos , Metabolismo/genética , Toxicologia/métodos , Universidades , Xenobióticos/toxicidade , Animais , Bases de Dados Factuais , Indústria Farmacêutica , Londres , Camundongos , Ressonância Magnética Nuclear Biomolecular , Ratos , Toxicologia/normas , Xenobióticos/sangue , Xenobióticos/urinaRESUMO
The role that metabonomics has in the evaluation of xenobiotic toxicity studies is presented here together with a brief summary of published studies. To provide a comprehensive assessment of this approach, the Consortium for Metabonomic Toxicology (COMET) has been formed between six pharmaceutical companies and Imperial College of Science, Technology and Medicine (IC), London, UK. The objective of this group is to define methodologies and to apply metabonomic data generated using (1)H NMR spectroscopy of urine and blood serum for preclinical toxicological screening of candidate drugs. This is being achieved by generating databases of results for a wide range of model toxins which serve as the raw material for computer-based expert systems for toxicity prediction. The project progress on the generation of comprehensive metabonomic databases and multivariate statistical models for prediction of toxicity, initially for liver and kidney toxicity in the rat and mouse, is reported. Additionally, both the analytical and biological variation which might arise through the use of metabonomics has been evaluated. An evaluation of intersite NMR analytical reproducibility has revealed a high degree of robustness. Second, a detailed comparison has been made of the ability of the six companies to provide consistent urine and serum samples using a study of the toxicity of hydrazine at two doses in the male rat, this study showing a high degree of consistency between samples from the various companies in terms of spectral patterns and biochemical composition. Differences between samples from the various companies were small compared to the biochemical effects of the toxin. A metabonomic model has been constructed for urine from control rats, enabling identification of outlier samples and the metabolic reasons for the deviation. Building on this success, and with the completion of studies on approximately 80 model toxins, first expert systems for prediction of liver and kidney toxicity have been generated.