Computational exploration of chemical space is crucial in modern cheminformatics research for accelerating the discovery of new biologically active compounds. In this study, we present a detailed analysis of the chemical library of potential glucocorticoid receptor (GR) ligands generated by the molecular generator, Molpher. To generate the targeted GR library and construct the classification models, structures from the ChEMBL database as well as from the internal IMG library, which was experimentally screened for biological activity in the primary luciferase reporter cell assay, were utilized. The composition of the targeted GR ligand library was compared with a reference library that randomly samples chemical space. A random forest model was used to determine the biological activity of ligands, incorporating its applicability domain using conformal prediction. It was demonstrated that the GR library is significantly enriched with GR ligands compared to the random library. Furthermore, a prospective analysis demonstrated that Molpher successfully designed compounds, which were subsequently experimentally confirmed to be active on the GR. A collection of 34 potential new GR ligands was also identified. Moreover, an important contribution of this study is the establishment of a comprehensive workflow for evaluating computationally generated ligands, particularly those with potential activity against targets that are challenging to dock.

• Klíčová slova
• chemical space, de novo design, glucocorticoid receptor, molecular generation,
• MeSH
• knihovny malých molekul * farmakologie   chemie   MeSH
• lidé MeSH
• ligandy MeSH
• receptory glukokortikoidů * metabolismus   chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• knihovny malých molekul * MeSH
• ligandy MeSH
• receptory glukokortikoidů * MeSH

The development of biologically active molecules based on molecular recognition is an attractive and challenging task in medicinal chemistry and the molecules that can activate/deactivate certain receptors are of great medical interest. In this contribution, selected pyrimidine/piperidine derivatives were synthesized and tested for the ability to activate/deactivate Aryl hydrocarbon receptor (AhR) and Glucocorticoid receptor (GR). Tested compounds are shown to activate the receptors but to much lesser extent than positive controls, dioxin and dexamethasone for Ahr and GR, respectively. However, some of them antagonized the positive controls action. Although further in vivo studies are needed to fully characterize the bioactivities of these compounds, the reported in vitro evidences demonstrate that they might be used as the modulators of AhR and GR activities.

• Klíčová slova
• Antagonists, Aryl hydrocarbon receptor, Biotransformation, Glucocorticoid receptor, Immunological response, Piperidines, Pyrimidines,
• MeSH
• buňky Hep G2 MeSH
• HeLa buňky MeSH
• lidé MeSH
• molekulární modely MeSH
• objevování léků MeSH
• piperidiny chemie   farmakologie   MeSH
• pyrimidiny chemie   farmakologie   MeSH
• receptory aromatických uhlovodíků agonisté   antagonisté a inhibitory   metabolismus   MeSH
• receptory glukokortikoidů agonisté   antagonisté a inhibitory   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• piperidiny MeSH
• pyrimidiny MeSH
• receptory aromatických uhlovodíků MeSH
• receptory glukokortikoidů MeSH

The effect of corticosterone injection and of acute and repeated stress on rat liver cytosol glucocorticoid receptor was studied to ascertain whether corticosterone-induced glucocorticoid receptor (GR) regulation also takes place in intact animals as it does in adrenalectomized ones. Adult male rats were exposed to six different stressors (swimming, 10 mg/kg histamine i.p., 500 mU/kg vasopressin s.c., heat, immobilization and cold) acutely or three times daily for 18 days (repeated stress). Each of the stressors applied acutely provoked a pronounced increase of plasma corticosterone with subsequent induction of hepatic tyrosine aminotransferase activity. Depletion of cytosol receptor was however only noticed after swimming and histamine injection. On the other hand, sustained hypersecretion of corticosterone evoked by repeated stress significantly reduced the number of GR in rat liver cytosol without any change in Kd. It is concluded that in the presence of intact adrenal glands cytosol receptors are more resistant to corticosterone-induced depletion than in their absence. Further, repeated stress causes down-regulation of GR in the liver, most probably by sustained corticosterone secretion, yet the effect of other stress factors cannot be excluded.

• MeSH
• cytosol metabolismus   MeSH
• down regulace MeSH
• fyziologický stres metabolismus   MeSH
• inbrední kmeny potkanů MeSH
• játra metabolismus   MeSH
• kortikosteron metabolismus   fyziologie   MeSH
• krysa rodu Rattus MeSH
• receptory glukokortikoidů metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kortikosteron MeSH
• receptory glukokortikoidů MeSH

UNLABELLED: We have investigated the involvement of glucocorticoid on methamphetamine (MA) induced hyperpyrexia using a bio-telemetric system. A significant level of hyperpyrexia was observed in MA administered rats. In contrast, increase of body temperature was suppressed by adrenalectomy or by the administration of RU-486, an antagonist of the glucocorticoid receptor. These data suggest that the glucocorticoid receptor may be involved in hyperpyrexia induced by MA. KEYWORDS: methamphetamine - hyperpyrexia - glucocorticoid - corticosterone.

• MeSH
• adrenalektomie MeSH
• horečka chemicky indukované   MeSH
• kortikosteron krev   MeSH
• krysa rodu Rattus MeSH
• methamfetamin otrava   MeSH
• mifepriston farmakologie   MeSH
• potkani Wistar MeSH
• receptory glukokortikoidů metabolismus   MeSH
• stimulanty centrálního nervového systému otrava   MeSH
• tělesná teplota účinky léků   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kortikosteron MeSH
• methamfetamin MeSH
• mifepriston MeSH
• receptory glukokortikoidů MeSH
• stimulanty centrálního nervového systému MeSH

The glucocorticoid receptor (GR) is an important player in the life of a cell. This is underlined by a cohort of protein and nucleic acid structures interacting with the GR. Among many issues surrounding GR activity that are under active investigation, the role of microtubules (MTs) is still unclear. This article aims to evaluate the ayes and noes in favor of microtubule importance and then form a hypothesis on their function in GR activity.

• MeSH
• biologické modely MeSH
• buněčný cyklus fyziologie   MeSH
• fosforylace MeSH
• glukokortikoidy fyziologie   MeSH
• hormony fyziologie   MeSH
• lidé MeSH
• mikrotubuly fyziologie   MeSH
• receptory glukokortikoidů genetika   fyziologie   MeSH
• regulace genové exprese MeSH
• signální transdukce fyziologie   MeSH
• transkripční faktory fyziologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• glukokortikoidy MeSH
• hormony MeSH
• receptory glukokortikoidů MeSH
• transkripční faktory MeSH

Benzimidazole drugs lansoprazole and omeprazole are used for treatment of various gastrointestinal pathologies. Both compounds cause drug-drug interactions because they activate aryl hydrocarbon receptor and induce CYP1A genes. In the current paper, we examined the effects of lansoprazole and omeprazole enantiomers on the expression of key drug-metabolizing enzyme CYP3A4 in human hepatocytes and human cancer cell lines. Lansoprazole enantiomers, but not omeprazole, were equipotent inducers of CYP3A4 mRNA in HepG2 cells. All forms (S-, R-, rac-) of lansoprazole and omeprazole induced CYP3A4 mRNA and protein in human hepatocytes. The quantitative profiles of CYP3A4 induction by individual forms of lansoprazole and omeprazole exerted enantiospecific patterns. Lansoprazole dose-dependently activated pregnane X receptor PXR in gene reporter assays, and slightly modulated rifampicin-inducible PXR activity, with similar potency for each enantiomer. Omeprazole dose-dependently activated PXR and inhibited rifampicin-inducible PXR activity. The effects of S-omeprazole were much stronger as compared to those of R-omeprazole. All forms of lansoprazole, but not omeprazole, slightly activated glucocorticoid receptor and augmented dexamethasone-induced GR transcriptional activity. Omeprazole and lansoprazole influenced basal and ligand inducible expression of tyrosine aminotransferase, a GR-target gene, in HepG2 cells and human hepatocytes. Overall, we demonstrate here that omeprazole and lansoprazole enantiomers induce CYP3A4 in HepG2 cells and human hepatocytes. The induction comprises differential interactions of omeprazole and lansoprazole with transcriptional regulators PXR and GR, and some of the effects were enantiospecific. The data presented here might be of toxicological and clinical importance, since the effects occurred in therapeutically relevant concentrations.

• MeSH
• buňky Hep G2 MeSH
• cytochrom P-450 CYP3A genetika   metabolismus   MeSH
• hepatocyty účinky léků   metabolismus   MeSH
• kultivované buňky MeSH
• lanzoprazol škodlivé účinky   farmakologie   MeSH
• lidé MeSH
• omeprazol škodlivé účinky   farmakologie   MeSH
• pregnanový X receptor MeSH
• protivředové látky škodlivé účinky   farmakologie   MeSH
• receptory glukokortikoidů metabolismus   MeSH
• steroidní receptory metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• CYP3A4 protein, human MeSH Prohlížeč
• cytochrom P-450 CYP3A MeSH
• lanzoprazol MeSH
• omeprazol MeSH
• pregnanový X receptor MeSH
• protivředové látky MeSH
• receptory glukokortikoidů MeSH
• steroidní receptory MeSH

Glucocorticoids are widely used drugs in human pharmacotherapy. There is an increasing demand for tools allowing detection of the ligands for glucocorticoid receptor (GR), with regard to pre-clinical drug testing and environmental applications. We constructed human luciferase reporter gene cell line AZ-GR derived from HeLa human cervix carcinoma cells, which were stably transfected with reporter plasmid containing three copies of glucorticoid response element (GRE) upstream of luciferase reporter gene. We isolated five dexamethasone-responsive clones, and we further characterized two most responsive ones (AZ-GR). Dose-response analyses were performed with 22 different natural and synthetic steroids and the values of EC(50) were calculated. AZ-GR cells displayed high specificity and sensitivity to glucocorticoids, very low responsiveness to mineralocorticoids, but no responsiveness to estrogens, gestagens or androgens. Time-course analyses revealed that AZ-GR cells allow detection of GR activators soon after 14 h of the treatment (6-10-fold induction by 100 nM dexamethasone). Functionality of AZ-GR cells was not affected with cryopreservation. Generated reporter gene cell lines fully maintained responsiveness to glucocorticoids for 32 days in the culture and over 16 passages without significant alterations. The sensitivity of the assay allows high throughput format using 96-well plates. Collectively, we present here glucocorticoid-responsive stable reporter gene cell line that allows high throughput, rapid, sensitive and selective detection of GR activators, with possible use in pre-clinical research and environmental applications.

• MeSH
• glukokortikoidy farmakologie   MeSH
• HeLa buňky MeSH
• lidé MeSH
• luciferasy genetika   MeSH
• receptory glukokortikoidů metabolismus   MeSH
• reportérové geny genetika   MeSH
• rychlé screeningové testy * MeSH
• steroidy farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• glukokortikoidy MeSH
• luciferasy MeSH
• receptory glukokortikoidů MeSH
• steroidy MeSH

Androgen and glucocorticoid receptors have been recently described as key players in processes related to prostate cancer and mainly androgen receptor's inactivation was shown as an effective way for the prostate cancer treatment. Unfortunately, androgen deprivation therapy usually loses its effectivity and the disease frequently progresses into castration-resistant prostate cancer with poor prognosis. The role of the glucocorticoid receptor is associated with the mechanism of resistance; therefore, pharmacological targeting of glucocorticoid receptor in combination with antiandrogen treatment was shown as an alternative approach in the prostate cancer treatment. We introduce here the synthesis of novel 17α- and/or 21-ester or carbamate derivatives of hydrocortisone and evaluation of their biological activity towards androgen and glucocorticoid receptors in different prostate cancer cell lines. A 17α-butyryloxy-21-(alkyl)carbamoyloxy derivative 14 was found to diminish the transcriptional activity of both receptors (in single-digit micromolar range), with comparable potency to enzalutamide towards the androgen receptor, but weaker potency compared to mifepristone towards the glucocorticoid receptor. Lead compound inhibited proliferation and the formation of cell colonies in both androgen and glucocortiocid receptors-positive prostate cancer cell lines in low micromolar concentrations. Candidate compound 14 showed to interact with both receptors in cells and inhibited the translocation of receptors to nucleus and their activation phoshorylation. Moreover, binding to receptor's ligand binding domains was assessed by molecular modelling. Lead compound also induced the accumulation of cells in G1 phase and its combination with enzalutamide was shown to be more effective than enzalutamide alone.

• Klíčová slova
• Androgen receptor, Castration resistant prostate cancer, Glucocorticoid receptor, Hydrocortisone derivatives, Prostate cancer, Transcriptional activity,
• MeSH
• androgenní receptory * genetika   chemie   MeSH
• androgeny farmakologie   MeSH
• antagonisté androgenů farmakologie   MeSH
• chemorezistence MeSH
• hydrokortison farmakologie   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory prostaty rezistentní na kastraci * farmakoterapie   metabolismus   MeSH
• nitrily farmakologie   MeSH
• receptory glukokortikoidů MeSH
• signální transdukce MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• androgenní receptory * MeSH
• androgeny MeSH
• antagonisté androgenů MeSH
• enzalutamide MeSH Prohlížeč
• hydrokortison MeSH
• nitrily MeSH
• receptory glukokortikoidů MeSH

Aryl hydrocarbon receptor (AhR) and glucocorticoid receptor (GR) play crucial role in the regulation of drug metabolizing enzymes and in many essential physiological processes. Cellular signaling by these receptors shares several functional and regulatory features. Here we investigated regulatory cross-talk between these two receptors. Human hepatoma cells (HepG2) were the model of choice. We analyzed the effects of dexamethasone (DEX) and dioxin (TCDD) on i) expression of AhR and GRalpha mRNAs; ii) levels of AhR and GR proteins; iii) transcriptional activities of AhR and GR in reporter assays; iv) 7-ethoxyresorufin-O-deethylase activity (EROD). We found that both DEX and TCDD affected AhR and GR mRNAs expression, proteins levels and transcriptional activities in HepG2 cells. These effects on cellular signaling by AhR and GR comprised up-/down-regulation of gene expression and ligand-dependent protein degradation. We conclude that interactive regulatory cross-talk between GR and AhR receptors in HepG2 cells defines possible implications in physiology and drug metabolism. Future research should be focused on the investigation of AhR-GR cross-talk in various normal human cells and tissues both in vitro and in vivo.

• MeSH
• cytochrom P-450 CYP1A1 metabolismus   MeSH
• dexamethason farmakologie   MeSH
• genetická transkripce účinky léků   MeSH
• glukokortikoidy farmakologie   MeSH
• hepatocelulární karcinom enzymologie   genetika   metabolismus   MeSH
• interakce mezi receptory a ligandy * MeSH
• lidé MeSH
• messenger RNA metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádory jater enzymologie   genetika   metabolismus   MeSH
• pilotní projekty MeSH
• polychlorované dibenzodioxiny farmakologie   MeSH
• receptory aromatických uhlovodíků agonisté   genetika   metabolismus   MeSH
• receptory glukokortikoidů agonisté   genetika   metabolismus   MeSH
• regulace genové exprese enzymů účinky léků   MeSH
• signální transdukce účinky léků   MeSH
• transfekce MeSH
• transkripční faktory bHLH MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• AHR protein, human MeSH Prohlížeč
• cytochrom P-450 CYP1A1 MeSH
• dexamethason MeSH
• glucocorticoid receptor alpha MeSH Prohlížeč
• glukokortikoidy MeSH
• messenger RNA MeSH
• NR3C1 protein, human MeSH Prohlížeč
• polychlorované dibenzodioxiny MeSH
• receptory aromatických uhlovodíků MeSH
• receptory glukokortikoidů MeSH
• transkripční faktory bHLH MeSH

We used 2,3,8,9-tetrachlorodibenzo-p-dioxin (TCDD) and dexamethasone (DEX) to examine their effects on aryl hydrocarbon receptor (AhR) and glucocorticoid receptor (GR) in HeLa cells. TCDD (5 nM), DEX (100 nM) and their combination down-regulated GR after 24 h. DEX reversed AhR mRNA increase and AhR protein decrease caused by TCDD. Since AhR-GR cross-talk occurs in cell-type and species-specific manner, the presented data may serve as the basis in the understanding of mechanisms underlying mutual interactions between AhR and GR.

• MeSH
• dexamethason farmakologie   MeSH
• down regulace účinky léků   MeSH
• glukokortikoidy farmakologie   MeSH
• HeLa buňky MeSH
• látky znečišťující životní prostředí toxicita   MeSH
• lidé MeSH
• messenger RNA účinky léků   metabolismus   MeSH
• nádory děložního čípku metabolismus   MeSH
• polychlorované dibenzodioxiny toxicita   MeSH
• receptory aromatických uhlovodíků účinky léků   metabolismus   MeSH
• receptory glukokortikoidů účinky léků   metabolismus   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• dexamethason MeSH
• glukokortikoidy MeSH
• látky znečišťující životní prostředí MeSH
• messenger RNA MeSH
• polychlorované dibenzodioxiny MeSH
• receptory aromatických uhlovodíků MeSH
• receptory glukokortikoidů MeSH