As organoids and organ-on-chip (OoC) systems move toward preclinical and clinical applications, there is an increased need for method validation. Using a liquid chromatography-mass spectrometry (LC-MS)-based approach, we developed a method for measuring small-molecule drugs and metabolites in the cell medium directly sampled from liver organoids/OoC systems. The LC-MS setup was coupled to an automatic filtration and filter flush system with online solid-phase extraction (SPE), allowing for robust and automated sample cleanup/analysis. For the matrix, rich in, e.g., protein, salts, and amino acids, no preinjection sample preparation steps (protein precipitation, SPE, etc.) were necessary. The approach was demonstrated with tolbutamide and its liver metabolite, 4-hydroxytolbutamide (4HT). The method was validated for analysis of cell media of human stem cell-derived liver organoids cultured in static conditions and on a microfluidic platform according to Food and Drug Administration (FDA) guidelines with regards to selectivity, matrix effects, accuracy, precision, etc. The system allows for hundreds of injections without replacing chromatography hardware. In summary, drug/metabolite analysis of organoids/OoCs can be performed robustly with minimal sample preparation.
- MeSH
- chromatografie kapalinová metody MeSH
- extrakce na pevné fázi MeSH
- hmotnostní spektrometrie metody MeSH
- játra * metabolismus MeSH
- kapalinová chromatografie-hmotnostní spektrometrie MeSH
- knihovny malých molekul analýza metabolismus chemie MeSH
- laboratoř na čipu MeSH
- léčivé přípravky metabolismus analýza MeSH
- lidé MeSH
- organoidy * metabolismus cytologie MeSH
- tolbutamid metabolismus analýza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- validační studie MeSH
In this review, research on the use of microalgae as an option for bioremediation purposes of pharmaceutical compounds is reported and discussed thoroughly. Pharmaceuticals have been detected in water bodies around the world, attracting attention towards the increasing potential risks to humans and aquatic biota. Unfortunately, pharmaceuticals have no regulatory standards for safe disposal in many countries. Despite the advances in new analytical techniques, the current wastewater treatment facilities in many countries are ineffective to remove the whole presence of pharmaceutical compounds and their metabolites. Though new methods are substantially effective, removal rates of drugs from wastewater make the cost-effectiveness ratio a not viable option. Therefore, the necessity for investigating and developing more adequate removal treatments with a higher efficiency rate and at a lower cost is mandatory. The present review highlights the algae-based removal strategies for bioremediation purposes, considering their pathway as well as the removal rate and efficiency of the microalgae species used in assays. We have critically reviewed both application of living and non-living microalgae biomass for bioremediation purposes considering the most commonly used microalgae species. In addition, the use of modified and immobilized microalgae biomass for the removal of pharmaceutical compounds from water was discussed. Furthermore, research considering various microalgal species and their potential use to detoxify organic and inorganic toxic compounds were well evaluated in the review. Further research is required to exploit the potential use of microalgae species as an option for the bioremediation of pharmaceuticals in water.
- MeSH
- biodegradace MeSH
- biomasa MeSH
- léčivé přípravky metabolismus MeSH
- lidé MeSH
- mikrořasy * metabolismus MeSH
- odpadní voda MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
V současné éře rozvíjející se precizní medicíny psychiatrie jako obor stále nedostatečně těží z rozšiřujících se technologických možností. Přitom disponuje tradicí průkopníka terapeutického monitorování léčiv (TDM) v medicíně a v současnosti má zpra- covány jedny z nejkomplexnějších doporučených postupů TDM mezi obory. Jejich progresivní vývoj však s sebou přinesl některé metodologické problémy, které vedou na jedné straně k opomíjení TDM v běžné klinické praxi a na druhé straně pak k jeho zbytečně mechanické aplikaci. Primárním cílem TDM není dosáhnout „doporučené“ koncentrace psychofarmaka v krvi, ale umožnit nalezení takového individuálního režimu podávání léčiva, který přinese pro pacienta minimum rizik a maximum profitu. Pro plnohodnotné využívání TDM v klinické praxi je důležité porozumět příležitostem i rizikům TDM a dokázat je aplikovat ve správnou chvíli správnému pacientovi se správným zadáním. Obsahem tohoto článku je seznámení se základními principy TDM, především farmakokinetickými základy, užívanými termíny, genezí doporučení, správnou aplikací metody a konečně s uplatněním výsledků v klinické praxi.
In the current era of evolving precision medicine, psychiatry as a field still does not sufficiently benefit from the expansion of technological opportunities. Yet, it has a tradition of pioneering therapeutic drug monitoring (TDM) in medicine and has developed some of the most comprehensive TDM guidelines among disciplines. However, its progressive development has brought with it some methodological problems that lead to the neglect of TDM in routine clinical practice on the one hand and its unnecessarily mechanical application on the other. The primary goal of TDM is not to achieve the „recommended“ blood concentration of a psychopharmaceutical, but to enable to find an individual drug regimen that will result in minimum risk and maximum benefit for the patient. To fully exploit TDM in clinical practice, it is important to understand the opportunities and risks of TDM and to be able to apply it at the right time to the right patient with the right task. The scope of this article is to introduce the basic principles of TDM, especially the pharmacokinetic basis, the terms used, the genesis of the recommendations, the correct application of the method and finally the application of the results in clinical practice.
- MeSH
- cesty eliminace léčiva MeSH
- farmakokinetika MeSH
- farmakologické účinky MeSH
- klinické laboratorní techniky metody MeSH
- léčivé přípravky analýza metabolismus MeSH
- lidé MeSH
- monitorování léčiv metody MeSH
- psychotropní léky * analýza farmakologie metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
- MeSH
- enzymy MeSH
- léčivé přípravky * metabolismus MeSH
- lékové interakce * MeSH
- lidé MeSH
- transferasy MeSH
- Check Tag
- lidé MeSH
Overcoming the skin barrier properties efficiently, temporarily, and safely for successful transdermal drug delivery remains a challenge. We synthesized three series of potential skin permeation enhancers derived from natural amino acid derivatives proline, 4-hydroxyproline, and pyrrolidone carboxylic acid, which is a component of natural moisturizing factor. Permeation studies using in vitro human skin identified dodecyl prolinates with N-acetyl, propionyl, and butyryl chains (Pro2, Pro3, and Pro4, respectively) as potent enhancers for model drugs theophylline and diclofenac. The proline derivatives were generally more active than 4-hydroxyprolines and pyrrolidone carboxylic acid derivatives. Pro2-4 had acceptable in vitro toxicities on 3T3 fibroblast and HaCaT cell lines with IC50 values in tens of μM. Infrared spectroscopy using the human stratum corneum revealed that these enhancers preferentially interacted with the skin barrier lipids and decreased the overall chain order without causing lipid extraction, while their effects on the stratum corneum protein structures were negligible. The impacts of Pro3 and Pro4 on an in vitro transepidermal water loss and skin electrical impedance were fully reversible. Thus, proline derivatives Pro3 and Pro4 have an advantageous combination of high enhancing potency, low cellular toxicity, and reversible action, which is important for their potential in vivo use as the skin barrier would quickly recover after the drug/enhancer administration is terminated.
- MeSH
- aplikace kožní MeSH
- hydroxyprolin metabolismus MeSH
- kožní absorpce * MeSH
- kůže metabolismus MeSH
- kyseliny karboxylové metabolismus MeSH
- léčivé přípravky metabolismus MeSH
- lidé MeSH
- organické látky metabolismus MeSH
- permeabilita MeSH
- prolin * metabolismus MeSH
- pyrrolidinony farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
One of the main contributors to pharmaceutical pollution of surface waters are non-steroidal anti-inflammatory drugs (NSAIDs) that contaminate the food chain and affect non-target water species. As there are not many studies focusing on toxic effects of NSAIDs on freshwater fish species and specially effects after dietary exposure, we selected rainbow trout (Oncorhynchus mykiss) as the ideal model to examine the impact of two NSAIDs - diclofenac (DCF) and ibuprofen (IBP). The aim of our study was to test toxicity of environmentally relevant concentrations of these drugs together with exposure doses of 100× higher, including their mixture; and to deepen knowledge about the mechanism of toxicity of these drugs. This study revealed kidneys as the most affected organ with hyalinosis, an increase in oxidative stress markers, and changes in gene expression of heat shock protein 70 to be signs of renal toxicity. Furthermore, hepatotoxicity was confirmed by histopathological analysis (i.e. dystrophy, congestion, and inflammatory cell increase), change in biochemical markers, increase in heat shock protein 70 mRNA, and by oxidative stress analysis. The gills were locally deformed and showed signs of inflammatory processes and necrotic areas. Given the increase in oxidative stress markers and heat shock protein 70 mRNA, severe impairment of oxygen transport may be one of the toxic pathways of NSAIDs. Regarding the microbiota, an overgrowth of Gram-positive species was detected; in particular, significant dysbiosis in the Fusobacteria/Firmicutes ratio was observed. In conclusion, the changes observed after dietary exposure to NSAIDs can influence the organism homeostasis, induce ROS production, potentiate inflammations, and cause gut dysbiosis. Even the environmentally relevant concentration of NSAIDs pose a risk to the aquatic ecosystem as it changed O. mykiss health parameters and we assume that the toxicity of NSAIDs manifests itself at the level of mitochondria and proteins.
- MeSH
- antiflogistika nesteroidní metabolismus MeSH
- biologické markery metabolismus MeSH
- chemické látky znečišťující vodu * metabolismus MeSH
- diklofenak metabolismus MeSH
- dysbióza MeSH
- ekosystém MeSH
- epidemický výskyt choroby MeSH
- ibuprofen metabolismus toxicita MeSH
- kyslík metabolismus MeSH
- léčivé přípravky metabolismus MeSH
- messenger RNA metabolismus MeSH
- Oncorhynchus mykiss * metabolismus MeSH
- oxidační stres MeSH
- proteiny tepelného šoku HSP70 metabolismus MeSH
- reaktivní formy kyslíku metabolismus MeSH
- střevní mikroflóra * MeSH
- voda metabolismus MeSH
- zánět chemicky indukované MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Administration of drugs by inhalation is mainly used to treat lung diseases and is being investigated as a possible route for systemic drug delivery. It offers several benefits, but it is also fraught with many difficulties. The lung is a complex organ with complicated physiology and specific pharmacokinetic processes. Therefore, the exposure and subsequently efficacy of a drug after inhalation is affected by a number of factors. In this review, we summarize the main variables that may affect drug fate after inhalation delivery, such as physicochemical properties of the drug, pulmonary clearance and metabolism, pathophysiological factors and inhalation device. Factors that have impact on pharmacokinetic processes need to be considered during development as their correct setting can lead to new effective inhaled drugs.
- MeSH
- aplikace inhalační MeSH
- léčivé přípravky metabolismus MeSH
- lidé MeSH
- plíce * metabolismus MeSH
- systémy cílené aplikace léků * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
The diffusion of biologically active molecules is a ubiquitous process, controlling many mechanisms and the characteristic time scales for pivotal processes in living cells. Here, we show how a high static magnetic field (MF) affects the diffusion of paramagnetic and diamagnetic species including oxygen, hemoglobin, and drugs. We derive and solve the equation describing diffusion of such biologically active molecules in the presence of an MF as well as reveal the underlying mechanism of the MF's effect on diffusion. We found that a high MF accelerates diffusion of diamagnetic species while slowing the diffusion of paramagnetic molecules in cell cytoplasm. When applied to oxygen and hemoglobin diffusion in red blood cells, our results suggest that an MF may significantly alter the gas exchange in an erythrocyte and cause swelling. Our prediction that the diffusion rate and characteristic time can be controlled by an MF opens new avenues for experimental studies foreseeing numerous biomedical applications.
Capillary electrophoresis-frontal analysis (CE-FA) together with mobility shift affinity CE is the most frequently used mode of affinity CE for a study of plasma protein-drug interactions, which is a substantial part of the early stage of drug discovery. Whereas in the classic CE-FA setup the sample is prepared by off-line mixing of the interaction partners in the sample vial outside the CE instrument and after a short incubation period loaded into the capillary and analysed, in this work a new methodological approach has been developed that combines CE-FA with the mixing of interacting partners directly inside the capillary. This combination gives rise to a fully automated and versatile methodology for the characterization of these binding interactions besides a substantial reduction in the amounts of sample compounds used. The minimization of possible experimental errors due to the full involving of sophisticated CE instrument in the injection procedure, mixing and separation instead of manual manipulation is another fundamental benefit. The in-capillary mixing is based on the transverse diffusion of laminar flow profile methodology introduced by Krylov et al. using its multi-zone injection modification presented by Řemínek at al.. Actually, after the method optimization, the alternate introduction of six plugs of drug and six plugs of bovine serum protein in BGE, each injected for 3 s at a pressure of -10 mbar (-1 kPa) into the capillary filled by BGE, was found to be the best injection procedure. The method repeatability calculated as RSDs of plateau highs of bovine serum albumin and propranolol as model sample compounds were better than 3.44 %. Its applicability was finally demonstrated on the determination of apparent binding parameters of bovine serum albumin for basic drugs propranolol and lidocaine and acid drug phenylbutazone. The values obtained by a new on-line CE-FA methodology are in agreement with values estimated by classic off-line CE-FA, as well as with literature data obtained using different techniques.
Although oral drug delivery is the preferred administration route and has been used for centuries, modern drug discovery and development pipelines challenge conventional formulation approaches and highlight the insufficient mechanistic understanding of processes critical to oral drug absorption. This review presents the opinion of UNGAP scientists on four key themes across the oral absorption landscape: (1) specific patient populations, (2) regional differences in the gastrointestinal tract, (3) advanced formulations and (4) food-drug interactions. The differences of oral absorption in pediatric and geriatric populations, the specific issues in colonic absorption, the formulation approaches for poorly water-soluble (small molecules) and poorly permeable (peptides, RNA etc.) drugs, as well as the vast realm of food effects, are some of the topics discussed in detail. The identified controversies and gaps in the current understanding of gastrointestinal absorption-related processes are used to create a roadmap for the future of oral drug absorption research.
- MeSH
- aplikace orální MeSH
- gastrointestinální trakt metabolismus MeSH
- interakce mezi potravou a léky MeSH
- intestinální absorpce * MeSH
- léčivé přípravky chemie metabolismus MeSH
- lidé MeSH
- počítačová simulace MeSH
- příprava léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
