Závěrečná zpráva o řešení grantu Agentury pro zdravotnický výzkum MZ ČR
Nestr.
Nespecifické střevní záněty (např. Crohnova choroba) jsou onemocnění se stále narůstající incidencí, jejichž současná farmakoterapie není zcela efektivní. Existuje ovšem celá řada přírodních aktivních látek s pozitivním vlivem při terapii střevních zánětů, které jsou intenzivně zkoumány. Prozatím ale nebyla testována možná terapeutická synergie mezi těmito látkami a glukany, přírodními sacharidy s dobře zdokumentovaným imunomodulačním účinkem. Glukanové částice mohou sloužit jako nosiče biologicky aktivních látek, což zjednoduší aplikaci ve vodě nerozpustných látek do organismu a zároveň skýtá možnost razantního zesílení protizánětlivého účinku. Na základě předchozích in vivo a klinických studií byly vytipovány chemicky různorodé přírodní látky, které jsou slibné pro terapii chronických zánětlivých onemocnění a lze je enkapsulovat do glukanových částic. Cílem projektu je pomocí in vitro a in vivo studií ověřit, zda je vhodné použít glukanové mikročástice jako nosiče těchto protizánětlivých látek k terapii střevních zánětů a optimalizovat složení kompozitu glukan-aktivní látka.; Inflammatory Bowel Diseases (IBD), e.g. Crohn’s Disease, are disorders with constantly increasing incidence and not completely effective pharmacotherapy. Number of active natural compounds with positive effect in therapy of IBD is intensively studied. Therapeutic synergy between them and glucans - natural saccharides with well-documented immunomodulatory effect - was not tested so far. Glucan microparticles can serve as carriers for biologically active compounds to simplify the administration of water insoluble drugs into organism and to open the possibility of significant amplification of anti-inflammatory effect as well. Based on previous in vivo and clinical trials, chemically diverse natural compounds, which may be encapsulated into glucan particles, were chosen as promising for therapy of chronic inflammatory disorders. The aim of project is to prove via in vitro and in vivo studies the suitability of utilization the glucan microparticles as vectors of these anti-inflammatory compounds in the therapy of IBD and to optimize the composition of glucan-active compound composite.
- MeSH
- antiflogistika MeSH
- biologické faktory MeSH
- glukany terapeutické užití MeSH
- idiopatické střevní záněty farmakoterapie MeSH
- intravitální mikroskopie MeSH
- lidé MeSH
- mikropartikule MeSH
- nosiče léků MeSH
- techniky in vitro MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- hodnotící studie MeSH
- Konspekt
- Farmacie. Farmakologie
- NLK Obory
- gastroenterologie
- farmacie a farmakologie
- NLK Publikační typ
- závěrečné zprávy o řešení grantu AZV MZ ČR
A series of sixteen ring-substituted N-arylcinnamanilides, previously described as highly antimicrobially effective against a wide spectrum of bacteria and fungi, together with two new derivatives from this group were prepared and characterized. Moreover, the molecular structure of (2E)-N-(2-bromo-5-fluorophenyl)-3-phenylprop-2-enamide as a model compound was determined using single-crystal X-ray analysis. All the compounds were tested for their anti-inflammatory potential, and most tested compounds significantly attenuated the lipopolysaccharide-induced NF-κB activation and were more potent than the parental cinnamic acid. (2E)-N-[2-Chloro-5-(trifluoromethyl)phenyl]-3-phenylprop-2-enamide, (2E)-N-(2,6-dibromophenyl)- 3-phenylprop-2-enamide, and (2E)-N-(2,5-dichlorophenyl)-3-phenylprop-2-enamide demonstrated the highest inhibition effect on transcription factor NF-κB at the concentration of 2 µM and showed a similar effectiveness as the reference drug prednisone. Several compounds also decreased the level of TNF-α. Nevertheless, subsequent tests showed that the investigated compounds affect neither IκBα level nor MAPKs activity, which suggests that the N-arylcinnamanilides may have a different mode of action to prednisone. The modification of the C(2,5)' or C(2,6)' positions of the anilide core by rather lipophilic and bulky moieties seems to be preferable for the anti-inflammatory potential of these compounds.
- MeSH
- antiflogistika chemická syntéza chemie farmakologie MeSH
- cinnamáty chemická syntéza chemie farmakologie MeSH
- krystalografie rentgenová MeSH
- lidé MeSH
- lipopolysacharidy škodlivé účinky MeSH
- molekulární modely MeSH
- molekulární struktura MeSH
- NF-kappa B metabolismus MeSH
- regulace genové exprese účinky léků MeSH
- signální transdukce účinky léků MeSH
- THP-1 buňky MeSH
- TNF-alfa metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Stilbenoids are important components of foods (e.g., peanuts, grapes, various edible berries), beverages (wine, white tea), and medicinal plants. Many publications have described the anti-inflammatory potential of stilbenoids, including the widely known trans-resveratrol and its analogues. However, comparatively little information is available regarding the activity of their prenylated derivatives. One new prenylated stilbenoid (2) was isolated from Artocarpus altilis and characterized structurally based on 1D and 2D NMR analysis and HRMS. Three other prenylated stilbenoids were prepared synthetically (9-11). Their antiphlogistic potential was determined by testing them together with known natural prenylated stilbenoids from Macaranga siamensis and Artocarpus heterophyllus in both cell-free and cell assays. The inhibition of 5-lipoxygenase (5-LOX) was also shown by simulated molecular docking for the most active stilbenoids in order to elucidate the mode of interaction between these compounds and the enzyme. Their effects on the pro-inflammatory nuclear factor-κB (NF-κB) and the activator protein 1 (AP-1) signaling pathway were also analyzed. The THP1-XBlue-MD2-CD14 cell line was used as a model for determining their anti-inflammatory potential, and lipopolysaccharide (LPS) stimulation of Toll-like receptor 4 induced a signaling cascade leading to the activation of NF-κB/AP-1. The ability of prenylated stilbenoids to attenuate the production of pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) was further evaluated using LPS-stimulated THP-1 macrophages.
- MeSH
- buněčné linie MeSH
- cyklooxygenasy metabolismus MeSH
- inhibitory enzymů farmakologie MeSH
- lidé MeSH
- lipoxygenasy metabolismus MeSH
- NF-kappa B antagonisté a inhibitory MeSH
- prenylace * MeSH
- signální transdukce účinky léků MeSH
- stilbeny farmakologie MeSH
- transkripční faktor AP-1 antagonisté a inhibitory MeSH
- zánět prevence a kontrola MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Quaternary benzophenanthridine alkaloids are known to have a wide range of biological effects, including antimicrobial, antifungal, anti-inflammatory, and antitumour activities. However, only sanguinarine and chelerythrine have been studied intensively. The aim of this study was to evaluate the anti-inflammatory potential of the five minor quaternary benzophenanthridine alkaloids sanguilutine, sanguirubine, chelirubine, chelilutine, and macarpine in vitro and to compare them with more thoroughly studied sanguinarine and chelerythrine. Before making cell-based assays, the cytotoxicity of the alkaloids was evaluated. The anti-inflammatory potential of the chosen alkaloids was evaluated as for their ability to modulate the lipopolysaccharide-induced secretion of tumour necrosis factor α (TNF-α) in the macrophage-like cell line THP-1. The cyclooxygenase (COX)-1 and COX-2 inhibitory activities were also measured. The results indicate that the presence of a methylenedioxy ring attached at carbon (C)7-C8 is important for reducing the secretion of TNF-α. Interestingly, this effect did not show a simple dependence on concentration. The selected alkaloids showed little or no anti-COX activity. The results obtained from the present experiments may provide additional information useful in understanding the structure-to-activity relationship of the quaternary benzophenanthridine alkaloids. The anti-inflammatory potential and the cytotoxic effect are driven by the presence of a methylenedioxy ring attached at C7-C8 and C2-C3, respectively.
A series of gold(I) triphenylphosphine (PPh3) complexes (1-9) involving 2-chloro-N6-(substituted-benzyl)adenine derivatives as N-donor ligands was synthesized and thoroughly characterized by relevant methods, including electrospray-ionization (ESI) mass spectrometry and multinuclear NMR spectroscopy. The anti-inflammatory and antiedematous effects of three representatives 1, 5 and 9 were evaluated by means of in vitro model based on the expression of pro- and anti-inflammatory cytokines and influence of the complexes on selected forms of matrix metalloproteinases secreted by LPS-activated THP-1 monocytes and in vivo model evaluating the antiedematous effect of the complexes in the carrageenan-induced rat hind-paw edema model. In addition to the pharmacological observations, the affected hind paws were post mortem subjected to histological and immunohistochemical evaluations. The results of both in vivo and ex vivo methods revealed low antiedematous and anti-inflammatory effects of the complexes, even though the in vitro model identified them as promising anti-inflammatory acting compounds. The reason for this discrepancy lies probably in low stability of the studied complexes in biological environment, as demonstrated by the solution interaction studies with sulfur-containing biomolecules (cysteine and reduced glutathione) using the ESI mass spectrometry.
- MeSH
- adenin chemie MeSH
- antiflogistika chemie farmakologie terapeutické užití MeSH
- buněčné linie MeSH
- cytokiny metabolismus MeSH
- edém farmakoterapie MeSH
- ELISA MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
- krysa rodu rattus MeSH
- ligandy MeSH
- organofosforové sloučeniny chemie farmakologie terapeutické užití MeSH
- potkani Wistar MeSH
- techniky in vitro MeSH
- zlato chemie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Kinetin (N6-furfuryladenine) belongs to a group of plant growth hormones involved in cell division, differentiation and other physiological processes. One of the possible ways to obtain biologically active compounds is to complex biologically relevant natural compounds to suitable metal atoms. In this work, two structural groups of Zn(II) complexes [Zn(L(n))2Cl2]·Solv (1-5) and [Zn(HL(n))Cl3] · xL(n) (6-7); n=1-5, Solv=CH3OH for 1 and 2H2O for 2; x =1 for 6 and 2 for 7; involving a phytohormone kinetin and its derivatives (L(n)) were evaluated for their ability to modulate secretion of tumour necrosis factor (TNF)-α, interleukin (IL)-1β and matrix metalloproteinase (MMP)-2 in a lipopolysaccharide (LPS)-activated macrophage-like THP-1 cell model. The penetration of the complexes to cells was also detected. The mechanism of interactions of the zinc(II) complexes with a fluorescent sensor N-(6-methoxy-8-quinolyl)-p-toluene sulphonamide (TSQ) and sulfur-containing biomolecules (l-cysteine and reduced glutathione) was studied by electrospray-ionization mass spectrometry and flow-injection analysis with fluorescence detection. The present study showed that the tested complexes exhibited a low cytotoxic effect on the THP-1 cell line (IC50>40 µM), apart from complex 4, with an IC50=10.9 ± 1.1 µM. Regarding the inflammation-related processes, the Zn(II) complexes significantly decreased IL-1β production by a factor of 1.47-2.22 compared with the control (DMSO), but did not affect TNF-α and MMP-2 secretions. However, application of the Zn(II) complexes noticeably changed the pro-MMP-2/MMP-2 ratio towards a higher amount of maturated MMP-2, when they induced a 4-times higher production of maturated MMP-2 in comparison with the vehicle-treated cells under LPS stimulation. These results indicated that the complexes are able to modulate an inflammatory response by influencing secretion and activity of several inflammation-related cytokines and enzymes.
- MeSH
- aktivace makrofágů účinky léků MeSH
- aminochinoliny MeSH
- antiflogistika chemická syntéza farmakologie MeSH
- biologický transport MeSH
- chloridy chemie MeSH
- cystein chemie MeSH
- exprese genu účinky léků MeSH
- fluorescenční barviva MeSH
- glutathion chemie MeSH
- interleukin-1beta antagonisté a inhibitory genetika sekrece MeSH
- kationty dvojmocné MeSH
- kinetin chemie MeSH
- komplexní sloučeniny chemická syntéza farmakologie MeSH
- lidé MeSH
- lipopolysacharidy farmakologie MeSH
- makrofágy cytologie účinky léků sekrece MeSH
- matrixová metaloproteinasa 2 genetika sekrece MeSH
- nádorové buněčné linie MeSH
- TNF-alfa genetika sekrece MeSH
- tosylové sloučeniny MeSH
- viabilita buněk účinky léků MeSH
- zinek chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
In this study, four prenylated and geranylated flavonoids, cudraflavone B (1), pomiferin (2), osajin (3), and diplacone (4), were tested for their antioxidant and anti-inflammatory effects and to identify any potential relationships between chemical structure and antioxidant or anti-inflammatory properties. The selected flavonoids were examined in cell-free models to prove their ability to scavenge superoxide radicals, hydrogen peroxide, and hypochlorous acid. Further, the ability of the flavonoids to influence the formation of reactive oxygen species in the murine macrophage cell line J774.A1 was tested in the presence and absence of lipopolysaccharide (LPS). The ability of flavonoids to inhibit LPS-induced IκB-α degradation and COX-2 expression was used as a model for the inflammatory response. The present results indicated that the antioxidant activity was dependent on the chemical structure, where the catechol moiety is especially crucial for this effect. The most potent antioxidant activities in cell-free models were observed for diplacone (4), whereas cudraflavone B (1) and osajin (3) showed a pro-oxidant effect in J774.A1 cells. All flavonoids tested were able to inhibit IκB-α degradation, but only diplacone (4) also down-regulated COX-2 expression.
- MeSH
- antiflogistika farmakologie MeSH
- antioxidancia farmakologie MeSH
- benzopyrany chemie farmakologie MeSH
- cyklooxygenasa 2 účinky léků MeSH
- flavonoidy chemie farmakologie MeSH
- inhibitory cyklooxygenasy farmakologie MeSH
- isoflavony chemie farmakologie MeSH
- kinasa I-kappa B antagonisté a inhibitory MeSH
- lipopolysacharidy farmakologie MeSH
- makrofágy účinky léků MeSH
- myši MeSH
- oxid dusnatý biosyntéza MeSH
- prenylace MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
