Colonic metabolism of ranitidine: implications for its delivery and absorption

The aim of this study was to assess the in vitro stability of ranitidine to colonic bacteria by utilising a batch culture fermentation system to simulate the conditions of the colon. Three quantities of ranitidine, 100, 200 and 500 mg, in the form of the hydrochloride salt, were introduced into individual 100 ml fermenters consisting of buffer medium inoculated with freshly voided human faeces (10% w/v). Control experiments were also run in parallel using equivalent drug quantities in buffer medium without the presence of faeces. Samples were removed at pre-determined time intervals over a 24 h period and were subsequently analysed by high-performance liquid chromatography (HPLC) for drug concentration. A selection of the samples removed from the fermenters was also analysed by conventional UV spectroscopy and mass spectrometry. Subsequent to an initial dissolution phase in the fermentation system, a marked decline in ranitidine concentration was noted over time, thereby suggesting degradation and metabolism of the drug by colonic bacteria. No such decline in concentration was noted in the control buffer systems. The rate and extent of metabolism was rapid and complete within 12 and 24 h for the 100 mg and 200 mg samples, respectively, although the largest sample size, 500 mg, was only partly metabolised over the course of the experiment. UV and mass spectrometry analysis indicated that metabolism occurred via cleavage of an N-oxide bond within the molecule with the resultant loss of an oxygen atom, although further metabolic reactions are possible. Such metabolism may in part be responsible for the poor bioavailability of ranitidine from the colon.     

Interactions between pesticides and components of pesticide formulations in an in vitro neurotoxicity test

Organophosphate (OP) pesticides are often used in combination with one another and with the components of formulations. Evidence already exists for interactions in the neurotoxic effects of OPs through interference with metabolism, but there is also potential for interactions related directly to cell damage. The purpose of this work was to investigate this possibility for OPs and the components of one of their common formulations in vitro. NB2a neuroblastoma cells were induced to differentiate in the presence of the OPs diazinon and chlorpyrifos, in combination with a commercial formulation (identified as Commercial Formulation 1) of the compounds and, independently, the components of that formulation. The compounds were tested in pairs in various proportions and the resulting inhibition of neurite outgrowth was measured by light microscopy and quantitative image analysis. Interactions were determined in terms of enhanced or reduced effects of the paired compounds in comparison with the expected additive effects estimated from the effects of each compound on its own. Synergism was detected between combinations of: 10 microM chlorpyrifos and 500 nM pyrethrum; chlorpyrifos and one of the solvents (regular spirit) found in Commercial Formulation 1. All other combinations of OPs and products were additive in their neurotoxicity. The data suggest that exposure to multiple OP-containing pesticide formulations may lead to synergistic neurotoxicity by a direct mechanism at the cellular level.     

Percutaneous absorption and metabolism of dinitrochlorobenzene in vitro

Dinitrochlorobenzene (DNCB) absorption through mouse and rat dorsal skin, pig ear skin and human abdominal skin in vitro was determined, and local metabolism to the glutathione conjugate was related to glutathione transferase activities and glutathione status in the skin. Absorption studies were conducted using skin mounted in a flow-through diffusion cell with tissue culture medium as receptor fluid. DNCB applied to the surface of skin in acetone penetrated through 26-day-old rat skin better than through the skin of the other species investigated. The amounts of absorption through pig and human skin and conjugation formation were similar. In general, occlusion resulted in increased penetration of DNCB but no change in conjugation. Human skin showed the highest gluta-thione-S-transferase activity towards DNCB, followed by 26-day-old rat, pig, mouse and neonatal rat skin. Levels of glutathione were highest in mouse skin, followed by neonatal rat, 26-day-old rat, pig and human skin, with pig and human skin showing similar levels. These studies indicated that the glutathione level in skin was the determining factor influencing the degree of DNCB conjugation during percutaneous absorption, and this was greatly depleted during percutaneous penetration of DNCB.     

Interactions between food components and drugs. Part 4: Influence of pectins and bile salts on propranolol absorption

Influence of dietary fiber components on drug absorption was studied in vitro using artificial membranes and mucosa preparations from guinea pig in 2-compartment model systems (permeation model and equilibrium dialysis). Well defined pectin preparations with different structural properties were used as food components and propranolol (P) as basic model drug. The retardation of drug was increased with decreased degree of esterification (DE) of pectin. Pectins with a blockwise distribution of free carboxyl groups possessed a more intensive effect than pectins with a random arrangement. It was found that P transport across the artificial lipid membrane was significantly decreased by pectins with a blockwise (DE <- 54%) or statistical (DE = 36%) distribution of free COOH. Pectins with lower molecular weight giving low viscosities in the medium showed only a small effect on permeation of the drug. Furthermore, the influence of bile acids without and with pectins on P absorption was studied. The bile salts did only influence P transport when they were applied above the critical micellar concentration (CMC). P transport across the lipid membranes increased slightly when pectins were additionally used to the bile salts above the CMC. Transport of P across the guinea pig mucosa was less than the permeation through the artificial lipid membranes. However, the transport of P across the mucosa was significantly reduced by glycocholic acid (GC), by pectin BL-3 as well as by BL-3 and GC in the same way as found using the artificial lipid membranes.     

Interactions of engineered nanomaterials in physiological media and implications for in vitro dosimetry

Abstract

In vitro toxicity assays are efficient and inexpensive tools for screening the increasing number of engineered nanomaterials (ENMs) entering the consumer market. However, the data produced by in vitro studies often vary substantially among different studies and from in vivo data. In part, these discrepancies may be attributable to lack of standardisation in dispersion protocols and inadequate characterisation of particle–media interactions which may affect the particle kinetics and the dose delivered to cells. In this study, a novel approach for preparation of monodisperse, stabilised liquid suspensions is presented and coupled with a numerical model which estimates delivered dose values. Empirically derived material- and media-specific functions are presented for each media–ENM system that can be used to convert administered doses to delivered doses. The interactions of ENMs with a variety of physiologic media were investigated and the importance of this approach was demonstrated by in vitro cytotoxicity assays using THP-1 macrophages.     

Interactions between cyclosporin and lipid-lowering drugs: implications for organ transplant recipients

Dyslipidaemia is more frequent in solid organ transplant recipients than in the general population, primarily as a result of immunosuppressive drug treatment. Both cyclosporin and corticosteroids are associated with dyslipidaemic adverse effects. In order to reduce the overall cardiovascular risk in these patients, lipid-lowering drugs have become widely used, especially HMG-CoA reductase inhibitors (statins). Cyclosporin, as well as most statins (lovastatin, simvastatin, atorvastatin and pravastatin) are metabolised by cytochrome P450 (CYP)3A4, so a bilateral pharmacokinetic interaction between these drugs is theoretically possible. However, results from several studies show that statins do not induce increased systemic exposure of cyclosporin. A small (but not clinically relevant) reduction in systemic exposure of cyclosporin has actually been shown in many studies. Cyclosporin-treated patients on the other hand show several-fold higher systemic exposure of all statins, both those that are metabolised by CYP3A4 and fluvastatin (metabolised by CYP2C9). Therefore, the mechanism for this interaction does not seem to be solely caused by inhibition of CYP3A4 metabolism, but it is probably also a result of inhibition of statin-transport in the liver, at least in part. Other lipid-lowering drugs, such as fibric acid derivatives, bile acid sequestrants, probucol, fish oils and orlistat are also used in solid organ transplant recipients. Most of them do not interact with cyclosporin, but there are reports indicating that both probucol and orlistat may reduce cyclosporin bioavailablility to a clinically relevant degree. There is no information on possible interaction effects of cyclosporin on the pharmacokinetics of lipid-lowering drugs other than statins, but it is not likely that any clinical relevant interference exists with fish oil, orlistat, probucol or bile acid sequestrants.     

Interactions between psychotropics, anaesthetics and electroconvulsive therapy: implications for drug choice and patient management

Despite many predictions that electroconvulsive therapy (ECT) would be replaced by pharmacotherapy, ECT has remained an invaluable adjunct in the management of severe psychiatric disease. Both pharmacotherapy and ECT continue to be used extensively, and will frequently be administered concurrently. The majority of patients requiring ECT will need anaesthesia; therefore, interactions could conceivably occur between the psychotropic drugs, ECT and the anaesthetic agents utilised. In managing an anaesthetic for ECT the effects of the anaesthetic agents and other medications on seizure intensity are important determinants influencing outcome. With regard to the antidepressants, tricyclic antidepressants (TCAs) and ECT can be combined safely and beneficially. More care is required when ECT is administered in the setting of a monoamine oxidase inhibitor (MAOI), especially the older irreversible varieties and in patients recently placed on MAOI therapy. Of the anticonvulsants and mood stabilisers, lithium and ECT given concurrently add significant risk of delirium and/or organic syndromes developing. Possible concerns with valproate, carbamazepine, lamotrigine, gabapentin and topiramate are that they may inhibit seizure activity. Additionally, carbamazepine may prolong the action of suxamethonium (succinylcholine). The combination of antipsychotics and ECT is well tolerated, and may in fact be beneficial. As regards the anxiolytics, benzodiazepines have anticonvulsant properties that might interfere with the therapeutic efficacy of ECT. CNS stimulants on the other hand may prolong seizures as well as produce dysrhythmias and elevate blood pressure. Calcium channel antagonists should be used with great care to avoid significant cardiovascular depression. The anaesthesiologist should therefore remain vigilant at all times, as untoward responses during ECT might occur suddenly due to interactions between psychotropics, anaesthetic agents and/or ECT.     

Interactions between corticotropin-releasing hormone and serotonin: implications for the aetiology and treatment of anxiety disorders

The amount of evidence for a role of aberrant serotoninergic neurotransmission in the aetiology of anxiety disorders, such as generalised anxiety and panic disorder, has been increasing steadily during the past several years. Although the picture is far from complete yet--partly due to the large number of serotonin (5-HT) receptors and the often-disparate effects of receptor agonists and antagonists in animal models of anxiety--SSRIs and the 5-HT1A agonist buspirone have now earned their place in the treatment of anxiety disorders. However, these drugs show--as they do in depressed patients--a delayed onset of improvement. Therefore, new therapeutical strategies are being explored. Corticotropin-releasing hormone (CRH), which plays a key role in the autonomic, neuroendocrine and behavioural responses to stress, is a strong anxiogenic neuropeptide and a promising candidate for therapeutical intervention in anxiety disorders. The neuroanatomical localisation of CRH, its congeners (the urocortins) and their receptors within the serotoninergic raphé nuclei suggests that interactions between the CRH system and 5-HT may play a role in fear and anxiety. In this chapter, I will discuss studies from my own and other laboratories showing that CRH and the urocortins influence several aspects of serotoninergic neurotransmission, including the firing rate of 5-HT neurones and the release and synthesis of this monoamine. Moreover, the interactions between CRH and 5-HT during psychologically stressful challenges will be discussed. Finally, I will review data showing that long-term alterations in the CRH system lead to aberrant functioning of serotoninergic neurotransmission under basal and/or stressful conditions. From this growing set of data the picture is emerging that the CRH system exerts a vast modulatory influence on 5-HT neurotransmission. An aberrant cross-talk between CRH and 5-HT may be of crucial importance in the neurobiology of anxiety disorders and represents, therefore, a promising goal for therapeutical intervention in these psychiatric diseases.     

Interactions between airway epithelial cells and dendritic cells: implications for the regulation of airway inflammation

Airway epithelial cells (AEC) and dendritic cells (DC) are situated in close proximity within the airway epithelium, and are the first cells to encounter inhaled pathogens, allergens and environmental pollutants. AEC and DC interact through the release of cytokines and other soluble mediators and through direct cell-cell contact, and these interactions are likely to play an important role in maintaining immune homeostasis. Increasing evidence indicates that both AEC and DC from asthmatic individuals exhibit distinct functional properties, compared with AEC and DC from healthy individuals. Both animal models, and novel co-culture models for directly studying human AEC/DC interactions are providing new insight into the cross-talk between these two important cells types, as a foundation for the development of new drug targets for use in asthma, cystic fibrosis and chronic obstructive pulmonary disease.     

Extrapolating In vitro Metabolic Interactions to Isolated Perfused Liver: Predictions of Metabolic Interactions between R-Bufuralol,Bunitrolol, and Debrisoquine

Drug-drug interactions (DDIs) are a great concern to the selection of new drug candidates. While in vitro screening assays for DDI are a routine procedure in preclinical research, their interpretation and relevance for the in vivo situation still represent a major challenge. The objective of the present study was to develop a novel mechanistic modeling approach to quantitatively predict DDI solely based upon in vitro data. The overall strategy consisted of developing a model of the liver with physiological details on three subcompartments: the sinusoidal space, the space of Disse, and the cellular matrix. The substrate and inhibitor concentrations available to the metabolizing enzyme were modeled with respect to time and were used to relate the in vitro inhibition constant (K_i) to the in vivo situation. The development of the liver model was supported by experimental studies in a stepwise fashion: (i) characterizing the interactions between the three selected drugs (R-bufuralol (BUF), bunitrolol (BUN), and debrisoquine (DBQ)) in microsomal incubations, (ii) modeling DDI based on binary mixtures model for all the possible pairs of interactions (BUF–BUN, BUF–DBQ, BUN–DBQ) describing a mutual competitive inhibition between the compounds, (iii) incorporating in the binary mixtures model the related constants determined in vitro for the inhibition, metabolism, transport, and partition coefficients of each compound, and (iv) validating the overall liver model for the prediction of the perfusate kinetics of each drug determined in isolated perfused rat liver (IPRL) for the single and paired compounds. Results from microsomal coincubations showed that competitive inhibition was the mechanism of interactions between all three compounds, as expected since those compounds are all substrates of rat CYP2D2. For each drug, the K values estimated were similar to their K_m values for CYP2D2 indicative of a competition for the same substrate-binding site. Comparison of the performance between the novel liver physiologically based pharmacokinetic (PBPK) model and published empirical models in simulating the perfusate concentration-time profile was based on the area under the curve (AUC) and the shape of the curve of the perfusate time course. The present liver PBPK model was able to quantitatively predict the metabolic interactions determined during the perfusions of mixtures of BUF-DBQ and BUN-DBQ. However, a lower degree of accuracy was obtained for the mixtures of BUF-BUN, potentially due to some interindividual variability in the relative proportion of CYP2D1 and CYP2D2 isoenzymes, both involved in BUF metabolism. Overall, in this metabolic interaction prediction exercise, the PBPK model clearly showed to be the best predictor of perfusate kinetics compared to more empirical models. The present study demonstrated the potential of the mechanistic liver model to enable predictions of metabolic DDI under in vivo condition solely from in vitro information.     

Differences in absorption, distribution, metabolism and excretion of xenobiotics between the paediatric and adult populations

In children, the therapeutic benefits and potential risks associated with drug treatment may be different from those in adults and will depend on the exposure, receptor sensitivity and relationship between effect and exposure. In this paper, key factors undergoing maturational changes accounting for differences in drug metabolism and disposition in the paediatric population compared with adults are reviewed. Gastric and duodenal pH, gastric emptying time, intestinal transit time, secretion and activity of bile and pancreatic fluid, bacterial colonisation and transporters, such as P-glycoprotein (P-gp), are important factors for drug absorption, whereas key factors explaining differences in drug distribution between the paediatric population and adults are organ size, membrane permeability, plasma protein concentration and characteristics, endogenous substances in plasma, total body and extracellular water, fat content, regional blood flow and transporters such as P-gp, which is present not only in the gut, but also in liver, kidney, brain and other tissues. As far as drug metabolism is concerned, important differences have been found in the paediatric population compared with adults both for phase I enzymes (oxidative [e.g., cytochrome P450 (CYP)1A2, and CYP3A7 versus -3A4], reductive and hydrolytic enzymes) and phase II enzymes (e.g., N-methyltransferases and glucuronosyltransferases). Generally, the major enzyme differences observed in comparison with the adult age are in newborn infants, although for some enzymes (e.g., glucuronosyltransferases and other phase II enzymes) important differences still exist between infants and toddlers and adults. Finally, key factors undergoing maturational changes accounting for differences in renal excretion in the paediatric population compared with adults are glomerular filtration and tubular secretion. The ranking of the key factors varies according to the chemical structure and physicochemical properties of the drug examined, as well as to the characteristics of its formulation. It would be important to generate additional information on the developmental aspects of renal P-gp and of other renal transporters, as has been done and is still being done with the different -isozymes involved in drug metabolism.     

Switching antiretroviral regimes for the treatment of HIV: safety implications

ABSTRACT

Introduction: There are multiple reasons to switch from a virologically successful antiretroviral regimen. Some of them are related to toxicity. Lately, combination antiretroviral treatment (cART) switches have often been related to drug-drug interactions which may also eventually entail safety issues as well.

Areas covered: The purpose of this review is to analyze causes of switching between virologically successful cART regimes related to safety issues. The most relevant papers were selected and summarized.

Expert opinion: Switching cART has been a popular strategy to address safety issues throughout the antiretroviral era. The myriad of switching studies have paralleled the study and release into clinical practice of new antiretroviral drugs with different and often improved safety profiles. Most of them have been successful in improving antiretroviral toxicity while keeping HIV replication under control. However, it should be taken into account that, whenever a new drug is given, there is a possibility of new drug-related toxicity. Notwithstanding that, an increase in cART switching is foreseen, given the fact that we have a wide antiretroviral drug armamentarium and that people living with HIV are ageing and thus more prone to developing age-related co-morbidities whose therapies may entail new interactions and eventually new toxicities.     

Epithelial transport of Noscapine across cell monolayer and influence of absorption enhancers on in vitro permeation and bioavailability: implications for intestinal absorption

Abstract

The purpose of this study was to investigate the permeation of Noscapine (Nos) across the Caco-2 and Madin–Darby canine kidney (MDCK) cell monolayers and to evaluate the influence of absorption enhancers on in vitro and in vivo absorption of Nos. The bidirectional transport of Nos was studied in Caco-2 and MDCK cell monolayers at pH 5.0–7.8. The effect of 0.5% w/v chitosan (CH) or Captisol (CP) on Nos permeability was investigated at pH 5.0 and 5.8. The effect of 1–5% w/v of CP on oral bioavailability of Nos (150?mg/kg) was evaluated in Sprague–Dawley rats. The effective permeability coefficients (P_eff) of Nos across Caco-2 and MDCK cell monolayers was found to be in the order of pH 5.0?>?5.8?>?6.8?>?7.8. The efflux ratios of P_eff?<?2 demonstrated that active efflux does not limit the absorption of Nos. The use of CH or CP have shown significant (***, p?<?0.001) enhancement in P_eff of Nos across cell monolayer compared with the control group. The CP (1–5% w/v) based Nos formulations resulted in significant (***, p?<?0.001) increase in the bioavailability of Nos compared with Nos solution. The use of CP represents viable approach for enhancing the oral bioavailability of Nos and reducing the required dose.     

Kinetic impact of presystemic intestinal metabolism on drug absorption: experiment and data analysis for the prediction of in vivo absorption from in vitro data

Orally administered drugs suffer from attack by metabolic enzymes not only in the liver, but also in the gastrointestine during the absorption process across the intestinal tissue. Although kinetic study on hepatic metabolism has been done well, the intestinal metabolism has not been well focused on compared with hepatic metabolism. In order to emphasize the role of intestinal metabolism in drug absorption and bioavailability, I have reviewed the experimental methods for intestinal absorption and metabolism, and the data analysis. Since Klippert et al. reported the prediction of intestinal first-pass effect of phenacetin in the rat from enzyme kinetic data in 1982, several reports have showed a good prediction, but others have not. Although intestinal absorption is an integrated process of transport (transporters) and metabolism (metabolic enzymes), most of the researchers missed the pathway of intestinal drug absorption and applied the kinetic model effective on only systemic metabolism to presystemic intestinal metabolism for their analysis of intestinal metabolism of orally administered drugs. A kinetic model, which incorporated factors of membrane transport, metabolic activity and protein binding, was structured to compare the equations in the reported models. In conclusion, we need more studies including kinetic modeling and experiments to understand the impact of intestinal metabolism on drug absorption. That knowledge must lead to the construction of ADME in silico (e-ADME).     

Interactions of C12 surfactants with the skin: studies on enzyme release and percutaneous absorption in vitro

Using an in vitro penetration cell, it has been shown that enzymes (acid phosphatase, lactate dehydrogenase and N-acetylglucosaminidase) are released from rat-skin slices in response to contact with two irritant C12 surfactants, sodium laurate and sodium lauryl sulphate, but not with the non-irritant sodium lauroyl isethionate. About 3-5 hr contact of the stratum corneum with surfactant and a long incubation time (24 hr) were required for enzyme release. Adsorption and penetration of the two effective surfactants was also studied and the results for sodium lauryl sulphate suggested a relationship between enzyme release and adsorption of surfactant. However, no such simple relationship was observed for sodium laurate, emphasizing the complex nature of surfactant interactions with the skin.     

In vitro metabolism of the fungicide and environmental contaminant trans-bromuconazole and implications for risk assessment

trans-Bromuconazole is a chiral chemical representative of a class of triazole derivatives known to inhibit specific fungal cytochrome P-450 (CYP) reactions. Kinetic measurements and delineation of metabolic pathways for triazole chemicals within in vitro hepatic microsomes are needed for accurate risk assessment and predictive in vivo physiological modeling. The studies described here were conducted with rat liver microsomes to determine Michaelis-Menten saturation kinetic parameters (Vmax and KM) for trans-bromuconazole using both substrate depletion and product formation reaction velocities. Kinetic parameters determined for trans-bromuconazole depletion at varying protein levels incubated at physiological temperature 37 degrees C resulted in a KM value of 1.69 microM and a Vmax value of 1398 pmol/min/mg protein. The concomitant linear formation of two metabolites identified using liquid chromatography/time-of-flight mass spectrometry (LC/MS-TOF) and LC-MS/MS indicated hydroxylation of the trans-bromuconazole dichlorophenyl ring moiety. KM values determined for the hydroxylated metabolites were 0.87 and 1.03 microM, with Vmax values of 449 and 694 pmol/min/mg protein, respectively. Chemical inhibition assays and studies conducted with individual purified human recombinant enzymes indicated the CYP3A subfamily was primarily responsible for biotransformation of the parent substrate. Additionally, trans-bromuconazole was found to undergo stereoselective metabolism as evidenced by a change in the enantiomeric ratio (trans-/trans+) with respect to time.