The human PXR (pregnane X receptor), a master regulator of drug metabolism, has essential roles in intestinal homeostasis and abrogating inflammation. Existing PXR ligands have substantial off-target toxicity. Based on prior work that established microbial (indole) metabolites as PXR ligands, we proposed microbial metabolite mimicry as a novel strategy for drug discovery that allows exploiting previously unexplored parts of chemical space. Here, we report functionalized indole derivatives as first-in-class non-cytotoxic PXR agonists as a proof of concept for microbial metabolite mimicry. The lead compound, FKK6 (Felix Kopp Kortagere 6), binds directly to PXR protein in solution, induces PXR-specific target gene expression in cells, human organoids, and mice. FKK6 significantly represses pro-inflammatory cytokine production cells and abrogates inflammation in mice expressing the human PXR gene. The development of FKK6 demonstrates for the first time that microbial metabolite mimicry is a viable strategy for drug discovery and opens the door to underexploited regions of chemical space.
- MeSH
- cytokiny MeSH
- kultivované buňky MeSH
- lidé MeSH
- ligandy MeSH
- molekulární mimikry * MeSH
- myši MeSH
- organoidy MeSH
- pregnanový X receptor chemie MeSH
- střeva MeSH
- zánět MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
Novel methylindoles were identified as endobiotic and xenobiotic ligands of the human aryl hydrocarbon receptor (AhR). We examined the effects of 22 methylated and methoxylated indoles on the transcriptional activity of AhRs. Employing reporter gene assays in AZ-AHR transgenic cells, we determined full agonist, partial agonist, or antagonist activities of tested compounds, having substantially variable EC50, IC50, and relative efficacies. The most effective agonists (EMAX relative to 5 nM dioxin) of the AhR were 4-Me-indole (134%), 6-Me-indole (91%), and 7-MeO-indole (80%), respectively. The most effective antagonists of the AhR included 3-Me-indole (IC50; 19 μM), 2,3-diMe-indole (IC50; 11 μM), and 2,3,7-triMe-indole (IC50; 12 μM). Reverse transcription polymerase chain reaction analyses of CYP1A1 mRNA in LS180 cells confirmed the data from gene reporter assays. The compound leads, 4-Me-indole and 7-MeO-indole, induced substantial nuclear translocation of the AhR and enriched binding of the AhR to the CYP1A1 promoter, as observed using fluorescent immunohistochemistry and chromatin immunoprecipitation assays, respectively. Molecular modeling and docking studies suggest the agonists and antagonists likely share the same binding pocket but have unique binding modes that code for their affinity. Binding pocket analysis further revealed that 4-methylindole and 7-methoxyindole can simultaneously bind to the pocket and produce synergistic interactions. Together, these data show a dependence on subtle and specific chemical indole structures as AhR modulators and furthermore underscore the importance of complete evaluation of indole compounds as nuclear receptor ligands.
- MeSH
- buňky Hep G2 MeSH
- cytochrom P-450 CYP1A1 metabolismus MeSH
- indoly farmakologie MeSH
- lidé MeSH
- ligandy MeSH
- messenger RNA metabolismus MeSH
- nádorové buněčné linie MeSH
- promotorové oblasti (genetika) účinky léků MeSH
- receptory aromatických uhlovodíků agonisté antagonisté a inhibitory MeSH
- reportérové geny účinky léků MeSH
- transkripční faktory bHLH agonisté antagonisté a inhibitory MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
Triazole antimycotic itraconazole contains in its structure three chiral centres; therefore, it forms eight stereoisomers. Commercial preparations of itraconazole are a mixture of four cis-diastereoisomers. There is much evidence that efficacy, adverse effects, and toxicity of chiral drugs may be stereospecific. Therefore, we have prepared 4 pure cis-diastereoisomers of itraconazole and investigated their effects on transcriptional activities of xenoreceptors aryl hydrocarbon receptor AhR and pregnane X receptor PXR. Gene reporter assays showed that itraconazole dose-dependently activated both AhR and PXR, and the activation of AhR but not of PXR was enantiospecific. Itraconazole diastereoisomers transformed AhR and PXR into their DNA-binding forms, as demonstrated by electromobility shift assays. Cytochrome P450 CYP1A1 mRNA and protein were induced by itraconazole diastereoisomers in human hepatoma cells HepG2, human skin cells HaCaT, and in primary human hepatocytes. The expression of CYP3A4 in human intestinal LS180 cells was not influenced by itraconazole, but we observed downregulation of CYP3A4 in human hepatocytes. Collectively, we show that itraconazole is a dual activator of AhR and PXR, with differential effects on the target genes for xenoreceptors. The enantiospecific pattern was observed only in gene reporter assays for AhR. The data presented here might be of toxicological and clinical importance.
- MeSH
- antifungální látky chemie farmakologie MeSH
- buněčné linie MeSH
- hepatocyty účinky léků metabolismus MeSH
- inhibitory cytochromu P450 CYP3A chemie farmakologie MeSH
- itrakonazol chemie farmakologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- messenger RNA metabolismus MeSH
- nádorové buněčné linie MeSH
- receptory aromatických uhlovodíků metabolismus MeSH
- stereoizomerie MeSH
- steroidní receptory metabolismus MeSH
- systém (enzymů) cytochromů P-450 genetika metabolismus MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Dihydropyridine calcium channel blockers (CCBs) are used as anti-hypertensives and in the treatment of angina pectoris. Structurally, CCBs have at least one chiral center in the molecule, thereby existing in two or more different enantiomers. In the current paper we examined effects of benidipine, felodipine and isradipine enantiomers on the expression and enzyme activities of human xenobiotics-metabolizing cytochromes P450. All CCBs dose-dependently activated aryl hydrocarbon receptor (AhR) and pregnane X receptor (PXR), as revealed by gene reporter assays. Activation of AhR, but not PXR, was enantiospecific. Consistently, CCBs induced CYP1A1 and CYP1A2 mRNAs, but not protein, in human hepatocytes and HepG2 cells, with following pattern: benidipine (-)>(+), isradipine (-)>(+) and felodipine (+)>(-). All CCBs induced CYP2A6, CYP2B6 and CYP3A4 mRNA and protein in human hepatocytes, and there were not differences between the enantiomers. All CCBs transformed AhR in its DNA-binding form, as revealed by electromobility shift assay. Tested CCBs inhibited enzyme activities of CYP3A4 (benidipine (+)>(-); felodipine (-)>(+); isradipine (-)-(+)) and CYP2C9 (benidipine (-)>(+); felodipine (+)>(-); isradipine (-)>(+)). The data presented here might be of toxicological and clinical importance.
- MeSH
- blokátory kalciových kanálů farmakologie MeSH
- buněčné linie MeSH
- dihydropyridiny chemie farmakologie MeSH
- hepatocyty účinky léků enzymologie MeSH
- izoenzymy metabolismus MeSH
- lidé MeSH
- messenger RNA biosyntéza genetika MeSH
- nádorové buněčné linie MeSH
- receptory aromatických uhlovodíků účinky léků MeSH
- stereoizomerie MeSH
- systém (enzymů) cytochromů P-450 metabolismus MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- vztahy mezi strukturou a aktivitou MeSH
- xenobiotika metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Manifestace diabetes mellitus 1. typu u HIV pacientů je celosvětově raritní komplikací tohoto základního onemocnění. Ve většině případů diabetu asociovaného s HIV se jedná o diabetes mellitus 2. typu. Inzulínová rezistence je v popředí patogenetického mechanismu rozvoje diabetu u těchto nemocných. Doposud bylo rozpoznáno mnoho rizikových faktorů a mnohé jsou stále ve stadiu výzkumu. Především zánětlivý stav podmíněný infekcí, délka trvání HIV infekce, nižší počet CD4+ lymfocytů, vyšší virová nálož, koinfekce virem hepatitidy C (HCV) a zejména léčba proteázovými inhibitory jsou hlavními z nich. Článek přináší kazuistiku pacienta a bližší seznámení s touto problematikou.
Manifestation of the type 1 diabetes mellitus in HIV patients is a rare complication of this underlying disease worldwide. In most cases of the HIV-associated diabetes it is the type 2 diabetes mellitus. The insulin resistance is a prominent factor of pathogenetic mechanism of the diabetes in these patients. Many risk factors have been diagnosed so far and many are still under the research. Firstly, the inflammatory condition caused by the infection, the duration of HIV infection, a decrease in the number of CD4+ lymphocytes, higher viral load, co-infection with hepatitis C virus (HCV) and particularly the treatment with protease inhibitor treatment are the major ones. The paper brings a patient’s case report and a closer familiarization with the topic.
- MeSH
- diabetes mellitus 1. typu * diagnóza epidemiologie etiologie patofyziologie MeSH
- dospělí MeSH
- Hepacivirus imunologie patogenita MeSH
- HIV infekce * diagnóza epidemiologie komplikace MeSH
- hyperglykemie farmakoterapie MeSH
- inzulinová rezistence imunologie MeSH
- lidé MeSH
- lymfadenitida epidemiologie etiologie MeSH
- věk při počátku nemoci * MeSH
- virová nálož imunologie MeSH
- vysoce aktivní antiretrovirová terapie metody škodlivé účinky MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
- Klíčová slova
- žádost o dotaci, občanská společnost,
- MeSH
- financování vládou * MeSH
- lidé MeSH
- neziskové organizace MeSH
- postižení * MeSH
- sociální péče MeSH
- zaměstnání pro postižené MeSH
- Check Tag
- lidé MeSH
- MeSH
- dekontaminace metody normy zákonodárství a právo MeSH
- indikátory a reagencie MeSH
- lidé MeSH
- směrnice jako téma MeSH
- statistika jako téma MeSH
- sterilizace metody normy využití MeSH
- vládní organizace MeSH
- zákonodárství jako téma MeSH
- zdravotnická zařízení normy zákonodárství a právo MeSH
- zdravotnický odpad klasifikace zákonodárství a právo MeSH
- Check Tag
- lidé MeSH
- Geografické názvy
- Česká republika MeSH
