The interaction of proton pump inhibitors with cytochromes P450.

An understanding of the role of human cytochromes (CYPs) P450 in the metabolism of proton pump inhibitors (PPIs) is important in assuring the safety of these drugs. Species' differences in the metabolism of PPIs tend to overestimate the risk of some forms of toxicity in humans. Therefore, information on metabolism by human enzymes is essential for the prediction of drug interactions, or the lack of them. Although extrapolation from population mean data to the individual patient should be made with care, available data indicate that the use of PPIs, omeprazole, lansoprazole and particularly pantoprazole, should be relatively free of clinically-significant drug interactions. This finding may reflect the level of therapeutic dosages for PPIs, and their short elimination half-lives with respect to exposure to drug- metabolizing enzymes. Concern about low-grade induction of CYPIA- mediated procarcinogen metabolism suggests a need for vigilance on the long-term use of PPIs.     

Pharmacogenetic diagnostics of cytochrome P450 polymorphisms in clinical drug development and in drug treatment

The current use and future perspectives of molecular genetic characterisation of cytochrome P450 enzymes (CYP) for drug development and drug treatment are summarised. CYP genes are highly polymorphic and the enzymes play a key role in the elimination of the majority of drugs from the human body. Frequent variants of some enzymes, CYP2A6, 2C9, 2C19 and 2D6, should be analysed in participants of clinical trials whenever these enzymes may play a role. It is suggested that a CYP genotype certificate is handed out to the volunteers or patients to avoid replicate analyses, and to allow that this information is available for future research and also for treatment with eventually needed drugs. Guidelines on what CYP alleles have to be analysed in drug development, as well as on analytical validation and CYP genotype data handling will be required. Treatment with several drugs may be improved by prior genotyping. The concepts and problems of CYP genotype-based clinical dose recommendations are presented and illustrated for selected drugs. The requirement for prospective trials on the medical and economic benefits of routine CYP genotyping is emphasised. Specific operationally defined recommendations dependent on genotype are a prerequisite for such studies and this review presents tentative CYP genotype-based dose recommendations systematically calculated from published data. Because of the multiplicity of factors involved, these doses will not be the optimal doses for each given individual, but should be more adequate than doses generally recommended for an average total population. Those CYP alleles and polymorphically metabolised drugs which are currently most interesting in drug development and drug treatment are reviewed, and more complete information is available from websites cited in this article.     

Roles of human cytochrome P450 enzymes involved in drug metabolism and toxicological studies

Multiple forms of cytochrome P450 (P450 or CYP) enzymes play important roles in the oxidation of structurally diverse xenobiotics and endobiotics. Interindividual variations in the level and activity of P450 enzymes were investigated in the human liver microsomes. Although the total P450 content was higher in Caucasian samples than in Japanese ones, the relative levels (percentage of total P450) of individual forms of P450 determined immunochemically were not very different. CYP3A (about 30% of total P450) and CYP2C (about 20%) enzymes were major forms. Different P450 enzymes in the human liver play major roles in a variety of drug oxidations and the hepatic contents of these P450 forms could be affective to determine which P450 enzymes play major roles in drug metabolism in individual humans. Recently recombinant P450 enzymes from different sources, e.g., microsomes of human lymphoblastoid cells, of yeast, and insect cells infected with baculovirus systems, and Escherichia coli membranes containing coexpressed P450 and reductase, have been widely used for drug metabolism research. However, the marker activities or kinetic parameters of human P450 enzymes reported are not always similar. Cytochrome b5 can enhance the activities of recombinant P450 systems in some cases using different mechanisms. These differences in activities may be a critical factor for understanding the roles of human P450 enzymes involved in drug metabolism. This review provides useful information for the study of drug biotransformation in humans and for the basis of drug toxicities and carcinogenesis.     

Optical substrates for drug-metabolizing enzymes: Recent advances and future perspectives

Drug-metabolizing enzymes (DMEs), a diverse group of enzymes responsible for the metabolic elimination of drugs and other xenobiotics, have been recognized as the critical determinants to drug safety and efficacy. Deciphering and understanding the key roles of individual DMEs in drug metabolism and toxicity, as well as characterizing the interactions of central DMEs with xenobiotics require reliable, practical and highly specific tools for sensing the activities of these enzymes in biological systems. In the last few decades, the scientists have developed a variety of optical substrates for sensing human DMEs, parts of them have been successfully used for studying target enzyme(s) in tissue preparations and living systems. Herein, molecular design principals and recent advances in the development and applications of optical substrates for human DMEs have been reviewed systematically. Furthermore, the challenges and future perspectives in this field are also highlighted. The presented information offers a group of practical approaches and imaging tools for sensing DMEs activities in complex biological systems, which strongly facilitates high-throughput screening the modulators of target DMEs and studies on drug/herb?drug interactions, as well as promotes the fundamental researches for exploring the relevance of DMEs to human diseases and drug treatment outcomes.     

Prediction of interindividual variation in drug plasma levels in vivo from individual enzyme kinetic data and physiologically based pharmacokinetic modeling.

A strategy is presented to predict interindividual variation in drug plasma levels in vivo by the use of physiologically based pharmacokinetic modeling and human in vitro metabolic parameters, obtained through the combined use of microsomes containing single cytochrome P450 enzymes and a human liver microsome bank. The strategy, applied to the pharmaceutical compound (N-[2-(7-methoxy-1-naphtyl)-ethyl]acetamide), consists of the following steps: (1) estimation of enzyme kinetic parameters K(m) and V(max) for the key cytochrome P450 enzymes using microsomes containing individual P450 enzymes; (2) scaling-up of the V(max) values for each individual cytochrome P450 involved using the ratio between marker substrate activities obtained from the same microsomes containing single P450 enzymes and a human liver microsome bank; (3) incorporation into a physiologically based pharmacokinetic model. For validation, predicted blood plasma levels and pharmacokinetic parameters were compared to those found in human volunteers: both the absolute plasma levels as well as the range in plasma levels were well predicted. Therefore, the presented strategy appears to be promising with respect to the integration of interindividual differences in metabolism and prediction of the possible impact on plasma and tissue concentrations of drugs in humans.     

Human membrane transporter database: A web-accessible relational database for drug transport studies and pharmacogenomics

The human genome contains numerous genes that encode membrane transporters and related proteins. For drug discovery, development, and targeting, one needs to know which transporters play a role in drug disposition and effects. Moreover, genetic polymorphisms in human membrane transporters may contribute to interindividual differences in the response to drugs. Pharmacogenetics, and, on a genome-wide basis, pharmacogenomics, address the effect of genetic variants on an individual's response to drugs and xenobiotics. However, our knowledge of the relevant transporters is limited at present. To facilitate the study of drug transporters on a broad scale, including the use of microarray technology, we have constructed a human membrane transporter database (HMTD). Even though it is still largely incomplete, the database contains information on more than 250 human membrane transporters, such as sequence, gene family, structure, function, substrate, tissue distribution, and genetic disorders associated with transporter polymorphisms. Readers are invited to submit additional data. Implemented as a relational database, HMTD supports complex biological queries. Accessible through a Web browser user interface via Common Gateway Interface (CGI) and Java Database Connection (JDBC) (http://128.218.208.23/transporter/trans.html), HMTD also provides useful links and references, allowing interactive searching and downloading of data. Taking advantage of the features of an electronic journal, this paper serves as an interactive tutorial for using the database, which we expect to develop into a research tool.     

Gallic acid and gallic acid derivatives: effects on drug metabolizing enzymes

Gallic acid and its structurally related compounds are found widely distributed in fruits and plants. Gallic acid, and its catechin derivatives are also present as one of the main phenolic components of both black and green tea. Esters of gallic acid have a diverse range of industrial uses, as antioxidants in food, in cosmetics and in the pharmaceutical industry. In addition, gallic acid is employed as a source material for inks, paints and colour developers. Studies utilising these compounds have found them to possess many potential therapeutic properties including anti-cancer and antimicrobial properties. In this review, studies of the effects of gallic acid, its esters, and gallic acid catechin derivatives on Phase I and Phase II enzymes are examined. Many published reports of the effects of the in vitro effects of gallic acid and its derivatives on drug metabolising enzymes concern effects directly on substrate (generally drug or mutagen) metabolism or indirectly through observed effects in Ames tests. In the case of the Ames test an antimutagenic effect may be observed through inhibition of CYP activation of indirectly acting mutagens and/or by scavenging of metabolically generated mutagenic electrophiles. There has been considerable interest in the in vivo effects of the gallate esters because of their incorporation into foodstuffs as antioxidants and in the catechin gallates with their potential role as chemoprotective agents. Principally an induction of Phase II enzymes has been observed however more recent studies using HepG2 cells and primary cultures of human hepatocytes provide evidence for the overall complexity of actions of individual components versus complex mixtures, such as those in food. Further systematic studies of mechanisms of induction and inhibition of drug metabolising enzymes by this group of compounds are warranted in the light of their distribution and consequent ingestion, current uses and suggested therapeutic potential. However, it must be noted that numerous constituents of foodstuffs have been found to be potent modulators of xenobiotic metabolism and the net human health effects may depend on concentrations of individual components and individual genetic makeup.     

Cytochrome P450 reaction-phenotyping: an industrial perspective

It is now widely accepted that the fraction of the dose metabolized by a given drug-metabolizing enzyme is one of the major factors governing the magnitude of a drug interaction and the impact of a polymorphism on (total) drug clearance. Therefore, most pharmaceutical companies determine the enzymes involved in the metabolism of a new chemical entity (NCE) in vitro, in conjunction with human data on absorption, distribution, metabolism and excretion. This so called reaction-phenotyping, or isozyme-mapping, usually involves the use of multiple reagents (e.g., recombinant proteins, liver subcellular fractions, enzyme-selective chemical inhibitors and antibodies). For the human CYPs, reagents are readily available and in vitro reaction-phenotyping data are now routinely included in most regulatory documents. Ideally, the various metabolites have been definitively identified, incubation conditions have afforded robust kinetic analyses, and well characterized (high quality) reagents and human tissues have been employed. It is also important that the various in vitro data are consistent (e.g., scaled turnover with recombinant CYP proteins, CYP inhibition and correlation data with human liver microsomes) and enable an integrated in vitro CYP reaction-phenotype. Results of the in vitro CYP reaction-phenotyping are integrated with clinical data (e.g., human radiolabel and drug interaction studies) and a complete package is then submitted for regulatory review. If the NCE receives market approval, information on key routes of clearance and their associated potential for drug-drug interactions are included in the product label. The present review focuses on in vitro CYP reaction-phenotyping and the integration of data. Relatively simple strategies enabling the design and prioritization of follow up clinical studies are also discussed.     

"The rock always comes first": drug users' accounts about using formal health care

Illicit drug use remains a significant public health threat. The issues surrounding drug use are recognized by public health professionals as important for several reasons. The incidence and prevalence of drug use persists in spite of the extensive societal, interpersonal, and individual consequences. In addition, the chronic health issues and health care costs associated with drug use continue to spiral. A wide variety of quantitative studies have examined the extent of health care problems, access, cost, and health care satisfaction among illicit drug users. While these studies offer important information through survey formats, fewer studies focus on subjective constructions of health care management from the users' perspective. This article examines the elements of the decision-making process involved in accessing formal health care among chronic and injecting street drug users. Twenty-eight in-depth interviews provide the data for this analysis, which is part of a large quantitative study of 1,479 injecting and chronic drug users and nondrug users in Miami, Florida. By exploring the elements of health care access through the eyes of the drug users, researchers and treatment professionals may gain insights into new ways to improve health care access for this at-risk population.     

Determination of a Human Hepatic Microsomal Scaling Factor for Predicting in Vivo Drug Clearance

Purpose To determine a microsomal scaling factor for human liver suitable for prediction of in vivo drug clearance from in vitro data and to explore the role of inter-liver variability in this factor on the reported underprediction from microsomal parameters. Methods Cytochrome P450 (henceforth P450) content in whole homogenates and microsomes from 38 donor livers was used to determine a microsomal scaling factor. In a subset (n = 20) of these preparations, individual P450 enzymes were examined by Western blotting and selective probe activities were determined. Results The scaling factor from 38 livers averaged 40 mg microsomal protein per gram liver with a coefficient of variation of 31%. Western blotting experiments indicated that there was no P450 enzyme-specific trend in the distribution of individual P450 enzymes in liver microsomes relative to whole homogenate. Predictions based on an average scaling factor resulted in a satisfactory prediction of intrinsic clearance of three benzodiazepines similar to that obtained using individual factors for the same livers. Conclusion A value for human liver microsomal scaling of 40 mg microsomal protein per gram liver has been established. The reason for underprediction previously reported for 52 different drug substrates was not the use of an incorrect value for the scaling factor.     

The use of multi-national web surveys for comparative analysis: Lessons from the European Web Survey on Drugs

BackgroundMost comparative drug policy analyses utilise measures of drug use, often from general population surveys (GPS). However, the limitations of GPS are well-recognised, including the small numbers of people who use illicit drugs sampled. Web surveys offer a potential solution to such issues. Therefore EMCDDA conducted a study to assess the potential for using such surveys to supplement information obtained from GPS.MethodsThe European Web Survey on Drugs (EWSD) asked about use of cannabis, amphetamines, cocaine and MDMA in 14 countries from 2016 to 2018. Each participant country translated the questionnaire as necessary and devised its own sampling strategy. Individuals aged 18+, resident in the participant country, who had used one or more of the drugs covered by the survey in the past 12 months were included in the analysis. Participation was anonymous and voluntary.ResultsMore than 40,000 people completed the survey, with recruitment mostly through social media. Larger samples of users of all drug types than found in GPS were generally obtained. However, the respondent profiles differed markedly between countries, e.g. the proportion aged 18–24 ranged from 30% to 80%. The results relating to use showed both inter-country similarities and differences, e.g. mean daily amounts of cocaine used varied between countries but increases in amounts used with increased frequency of use were similar. Price data showed good external validity.ConclusionWeb surveys offer the possibility of collecting information from large numbers people who use illicit drugs quickly and cheaply and can fill important gaps in our knowledge of patterns of use, particularly by recreational users. However, they also have limitations. Standardising questionnaires and approaches to data cleaning and analysis facilitates comparisons between countries but obtaining comparable samples may be challenging. Multinational surveys need to balance standardisation of methods with responsiveness to differing country contexts; our collaborative model does this.     

Bioinformatics of proteases in the MEROPS database

Proteolytic enzymes represent approximately approximately 2% of the total number of proteins present in all types of organisms. Many of these enzymes are of medical importance, and those that are of potential interest as drug targets can be divided into the endogenous enzymes encoded in the human genome, and the exogenous proteases encoded in the genomes of disease-causing organisms. There are also naturally occurring inhibitors of proteases, some of which have pharmaceutical relevance. The MEROPS database provides a rich source of information on proteases and their inhibitors. Storage and retrieval of this information is facilitated by the use of a hierarchical classification system (which was pioneered by the compilers of the database) in which homologous proteases and their inhibitors are divided into clans and families.     

Predicting drug clearance from recombinantly expressed CYPs: intersystem extrapolation factors

1. Recombinantly expressed human cytochromes P450 (rhCYPs) have been underused for the prediction of human drug clearance (CL). 2. Differences in intrinsic activity (per unit CYP) between rhCYP and human liver enzymes complicate the issue and these discrepancies have not been investigated systematically. We define intersystem extrapolation factors (ISEFs) that allow the use of rhCYP data for the in vitro-in vivo extrapolation of human drug CL and the variance that is associated with interindividual variation of CYP abundance due to genetic and environmental effects. 3. A large database (n = 451) of metabolic stability data has been compiled and used to derive ISEFs for the most commonly used expression systems and CYP enzymes. 4. Statistical models were constructed for the ISEFs to determine major covariates in order to optimize experimental design to increase prediction accuracy. 5. Suggestions have been made for the conduct of future studies using rhCYP to predict human drug clearance.     

Lived fictions: moving from pharmacology to ethnography

While positivist science dominates the drug and alcohol research sector, alternative constructivist paradigms of inquiry into drug and alcohol use are also present. The passage of individual researchers through paradigm shifts is often difficult, and rarely documented. Different paradigms of inquiry produce different types of information. The use of the psychoactive drug methylenedioxymethamphetamine (MDMA or “Ecstasy”) is discussed in light of a multidisciplinary investigation into the pharmacology and use of MDMA. The role of the author in interpreting data from both positivist and constructivist paradigms is highlighted by documenting a researchers' changes in epistemology whilst undertaking multidisciplinary research. It is concluded that multidisciplinary research into drug and alcohol use could benefit from evaluation of the processes that frame how information from both research paradigms is interpreted and integrated.     

Cytochrome P450 polymorphisms and drug-induced interstitial lung disease

INTRODUCTION: With an increasing number of therapeutic drugs available for use, the list of drugs that are responsible for severe pulmonary disease also grows. Genetic polymorphism of drug-metabolizing enzymes, particularly of the cytochrome P450 superfamily of enzymes, influences individual drug efficacy and safety through the alteration of pharmacokinetics and disposition of drugs. AREAS COVERED: This review focuses on drug-induced interstitial lung disease, describes common patterns of pulmonary injury, discusses diagnosis and treatment, and details the prevalence and clinical significance of cytochrome P450 polymorphisms. EXPERT OPINION: Polymorphisms of cytochrome P450 genes can influence the metabolic activity of the subsequent enzymes, which in turn may lead to localized reactions and tissue damage, for example, in lung tissue. Pharmacogenomic techniques allow efficient analysis of risk factors and genotyping tests have the potential to optimize drug therapy. In the future, genotyping should be considered to identify patients who are at high risk of severe toxic responses in order to guide appropriate individual dosage.     

Metabolism of antiarrhythmics

Antiarrhythmics are a group of drugs that manage the irregular electrical activity of the heart. Their use in the clinic is made difficult by their narrow therapeutic index. The disposition of antiarrhythmics is dependent on many factors, such as administration route, stereoselectivity in the first-pass effect, inhibition of enzymes, polymorphisms, etc. Consequently, the pharmacological activity of drugs may be interindividually variable. Experiments using organ homogenates or hepatic microsome fractions were used for simulating the biotransformation of the drug in vivo. The classical approaches, such as correlation analysis, specifically the inhibitory effect, or induction of chemicals, and immunoinhibition, may be combined with the use of recombinant enzymes for identifying the enzymes involved in the drug metabolism. The fate of the antiarrhythmics may also be investigated in live animals. A species-dependent metabolism was often observed. The pre-treatment with chemicals, which influences the change (inhibition or induction) in the drug disposition, may provide insights into the enzymes involved in vivo. However, published data indicated that the data obtained from animals should not be extrapolated directly to humans. Nevertheless, animal models are useful for investigating the mechanism of clinical observations. The clinical use of the antiarrhythmics becomes complex, when the drug metabolism is genetically/phenotypically dependent and active metabolites are formed. Furthermore, the stereoselectivity may also modify the disposition and the pharmacodynamic profile of a therapeutic agent. Only the knowledge of the drug metabolism and the status of each individual may allow the use of antiarrhythmics safely.     

The value of nonhuman primates in drug abuse research

The use of nonhuman primates (NHP) is invaluable for drug abuse research. The laboratory animals most closely related to humans are NHP. The phylogeny, anatomy, physiology, neurochemistry, and behavior of NHP are more similar to humans than other laboratory species. There is now an extensive body of literature documenting the neuroanatomical, neurochemical, and neuropharmacological similarities between NHP and humans and the differences between NHP and other laboratory species in dopamine, norepinephrine, serotonin, opioid, and gamma aminobutyric acid systems. Comprehensive studies comparing pharmacokinetics in humans, monkeys, dogs, and rats have shown that data in monkeys are the most predictive of human pharmacokinetic parameters. The long life span and extended adolescent period for NHP permits intensive, long-term investigations and the use of within-subject experimental designs similar to those used in human laboratory studies. Within-subject designs require fewer subjects than standard between-group designs and permit the careful evaluation of individual differences. NHP have been used extensively in drug abuse research for over 40 years and have provided useful information on the behavioral processes associated with drug abuse and addiction as well as drug abuse liability in humans. This review focuses on important species differences between rodents and NHP and on the value of NHP in bridging the gap between rodents and humans to enhance the ability to generalize preclinical findings to human drug abuse.