Trilobolide and its analogues belong to the guaianolide type of sesquiterpene lactones, which are characteristic and widely distributed within the families Asteraceae and Apiaceae. Certain guaianolides are receiving continuously increasing attention for their promising sarco-endoplasmic reticulum Ca2+-ATPase (SERCA)-inhibitory activity. However, because of their alkylation capabilities, they are generally toxic. Therefore, the search for compounds with significant immunobiological properties but with decreased cytotoxicities suitable for use in immune-based pharmacotherapy is ongoing. Therefore, we extended our previous investigation of the immunobiological effects of trilobolide to a series of structurally related guaianolides and germacranolides. To evaluate the relationship, we tested a series of selected derivatives containing α-methyl lactone or exomethylene lactone ring. For a wider comparison, we also included some of their glycosidic derivatives. We assessed the in vitro immunobiological effects of the tested compounds on nitric oxide (NO) production, cytokine secretion, and prostaglandin E2 (PGE2) release by mouse peritoneal cells, activated primarily by lipopolysaccharide (LPS), and evaluated their viability. The inhibitory effects of the apparently most active substance, 8-deoxylactucin, seem to be the most promising.
- Klíčová slova
- 8-deoxylactucin, 8-epiisoamberboin, germacranolides, guaianolides, immune-modulatory effects,
- MeSH
- butyráty MeSH
- cytokiny metabolismus MeSH
- dinoproston metabolismus biosyntéza MeSH
- furany MeSH
- laktony * farmakologie chemie MeSH
- lipopolysacharidy farmakologie MeSH
- myši MeSH
- oxid dusnatý * metabolismus MeSH
- peritoneální makrofágy účinky léků metabolismus MeSH
- seskviterpeny germakranové * farmakologie chemie MeSH
- seskviterpeny guajanové * farmakologie chemie MeSH
- seskviterpeny farmakologie chemie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- butyráty MeSH
- cytokiny MeSH
- dinoproston MeSH
- furany MeSH
- germacranolide MeSH Prohlížeč
- laktony * MeSH
- lipopolysacharidy MeSH
- oxid dusnatý * MeSH
- seskviterpeny germakranové * MeSH
- seskviterpeny guajanové * MeSH
- seskviterpeny MeSH
- trilobolide MeSH Prohlížeč
Saponins, a diverse group of natural compounds, offer an interesting pool of derivatives with biomedical application. In this study, three structurally related spirostanol saponins were isolated and identified from the leek flowers of Allium porrum L. (garden leek). Two of them were identical with the already known leek plant constituents: aginoside (1) and 6-deoxyaginoside (2). The third one was identified as new component of A. porrum; however, it was found identical with yayoisaponin A (3) obtained earlier from a mutant of elephant garlic Allium ampeloprasun L. It is a derivative of the aginoside (1) with additional glucose in its glycosidic chain, identified by MS and NMR analysis as (2α, 3β, 6β, 25R)-2,6-dihydroxyspirostan-3-yl β-D-glucopyranosyl-(1 → 3)-β-D-glucopranosyl-(1 → 2)-[β-D-xylopyranosyl-(1 → 3)]-β-D-glucopyranosyl]-(1 → 4)-β-D-galactopyranoside, previously reported also under the name alliporin. The leek native saponins were tested together with other known and structurally related saponins (tomatonin and digitonin) and with their related aglycones (agigenin and diosgenin) for in vitro cytotoxicity and for effects on NO production in mouse peritoneal cells. The highest inhibitory effects were exhibited by 6-deoxyaginoside. The obtained toxicity data, however, closely correlated with the suppression of NO production. Therefore, an unambiguous linking of obtained bioactivities of saponins with their expected immunobiological properties remained uncertain.
- Klíčová slova
- Allium porrum, NO production, aginoside, alliporin, cytotoxicity, leek flowers, steroid saponins,
- MeSH
- Allium chemie MeSH
- buněčné linie MeSH
- květy chemie MeSH
- lipopolysacharidy antagonisté a inhibitory farmakologie MeSH
- molekulární konformace MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- oxid dusnatý antagonisté a inhibitory biosyntéza MeSH
- peritoneální makrofágy účinky léků metabolismus MeSH
- saponiny chemie izolace a purifikace farmakologie MeSH
- spirostany chemie izolace a purifikace farmakologie MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- lipopolysacharidy MeSH
- oxid dusnatý MeSH
- saponiny MeSH
- spirostany MeSH
Sesquiterpene lactones are secondary plant metabolites with sundry biological effects. In plants, they are synthesized, among others, for pesticidal and antimicrobial effects. Two such compounds, archangelolide and trilobolide of the guaianolide type, are structurally similar to the well-known and clinically tested lactone thapsigargin. While trilobolide has already been studied by us and others, there are only scarce reports on the biological activity of archangelolide. Here we present the preparation of its fluorescent derivative based on a dansyl moiety using azide-alkyne Huisgen cycloaddition having obtained the two sesquiterpene lactones from the seeds of Laserpitium archangelica Wulfen using supercritical CO2 extraction. We show that dansyl-archangelolide localizes in the endoplasmic reticulum of living cells similarly to trilobolide; localization in mitochondria was also detected. This led us to a more detailed study of the anticancer potential of archangelolide. Interestingly, we found that neither archangelolide nor its dansyl conjugate did exhibit cytotoxic effects in contrast to the structurally closely related counterparts trilobolide and thapsigargin. We explain this observation by a molecular dynamics simulation, in which, in contrast to trilobolide, archangelolide did not bind into the sarco/endoplasmic reticular calcium ATPase cavity utilized by thapsigargin. Last, but not least, archangelolide exhibited anti-inflammatory activity, which makes it promising compound for medicinal purposes.
- MeSH
- Apiaceae chemie MeSH
- butyráty chemie farmakologie MeSH
- cytokiny imunologie MeSH
- furany chemie farmakologie MeSH
- kultivované buňky MeSH
- lidé MeSH
- molekulární struktura MeSH
- myši MeSH
- oxid dusnatý imunologie MeSH
- peritoneum cytologie MeSH
- potkani Wistar MeSH
- semena rostlinná chemie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- butyráty MeSH
- cytokiny MeSH
- furany MeSH
- oxid dusnatý MeSH
- trilobolide MeSH Prohlížeč
In this study we investigated the effect of five therapeutically used drugs and four natural polyphenolic compounds on the mechanism of oxidative burst of human neutrophils concerning their participation in the generation of reactive oxygen species (ROS). The compounds investigated decreased the oxidative burst of whole blood in the rank order of potency: N-feruloylserotonin > quercetin > curcumin > arbutin > dithiaden > carvedilol. The generation of intracellular reactive oxygen species in isolated neutrophils decreased in the same rank order, while carvedilol was ineffective. Scavenging of extracellular oxygen radicals followed the rank order of potency: N-feruloylserotonin > curcumin > quercetin > dithiaden. Arbutin and carvedilol had no effect. All compounds tested increased the activity of caspase-3 in cell-free system indicating a positive effect on apoptosis of neutrophils. Activation of protein kinase C was significantly decreased by dithiaden, curcumin, quercetin and N-feruloylserotonin. Carvedilol, dithiaden, quercetin and arbutin reduced activated neutrophil myeloperoxidase release more significantly compared with their less pronounced effect on superoxide generation The presented results are indicative of pharmacological intervention with neutrophils in pathological processes. Of particular interest was the effect of natural compounds. Intracellular inhibition of oxidative burst in isolated neutrophils by the drugs tested and natural antioxidants has to be further analysed since ROS play an important role in immunological responses of neutrophils.
- Klíčová slova
- human neutrophils, natural antioxidants, oxidative burst, therapeutical drugs,
- Publikační typ
- časopisecké články MeSH
Sesquiterpene lactone trilobolide is a sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) inhibitor, thus depleting the Ins(1,4,5)P3-sensitive intracellular calcium stores. Here, we describe a synthesis of a series of 6 trilobolide-steroids conjugates (estradiol, pregnene, dehydroepiandrosterone, and testosterone). We found that the newly synthesized Tb-based compounds possess different remarkable biological activities. Cancer cell cytotoxicity and preferential selectivity is represented in our study by a Tb-pregnene derivative. The most cytotoxic clickates of estradiol and pregnene were studied by FACS where impact on cell cycle and RNA synthesis was observed; live-cell microscopy revealed the impact on cell organelle morphology particularly endoplasmic reticulum, mitochondria and nucleus. Further, we have studied the estrogenic and androgenic properties of the clickate molecules using cell-based luciferase assays. Finally, antimycobacterial tests revealed that testosterone and estradiol derivatives potentiated the antimycobacterial activity up to IC50 of 10.6μM.
- Klíčová slova
- Click chemistry, Cytotoxicity, SAR, Steroid receptor, Steroids, Trilobolide,
- MeSH
- androgenní receptory metabolismus MeSH
- antibakteriální látky chemická syntéza chemie farmakologie MeSH
- buňky A549 MeSH
- butyráty chemie MeSH
- Candida účinky léků MeSH
- furany chemie MeSH
- HCT116 buňky MeSH
- lidé MeSH
- molekulární struktura MeSH
- nádorové buněčné linie MeSH
- receptory pro estrogeny metabolismus MeSH
- steroidní receptory metabolismus MeSH
- steroidy chemie MeSH
- syntetická chemie okamžité shody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- androgenní receptory MeSH
- antibakteriální látky MeSH
- butyráty MeSH
- furany MeSH
- receptory pro estrogeny MeSH
- steroidní receptory MeSH
- steroidy MeSH
- trilobolide MeSH Prohlížeč
Trilobolide (Tb) is a pharmacologically interesting sesquiterpene lactone isolated from Laser trilobum (L.) Borkh. Structural relation to a sarco/endoplasmic reticulum Ca2+-ATPase inhibitor thapsigargin bring promising prospects for Tb to be used in the development of new anti-cancer drugs. As long as there are still unanswered questions regarding its investigation, a need for novel analytical tools emerge. Since immunoassays serve as one of powerful tools within the investigation of natural products, the development of indirect competitive enzyme-linked immunosorbent assay (ELISA) utilizing coating based on avidin-biotin technology is described. In our set-up of ELISA, newly synthesized biotinylated Tb served as immobilized competitor. Tb-carboxymethyloxime-bovine serum albumin (BSA) and Tb-succinoyl-BSA conjugates were used separately for immunization of rabbits. Two sets of polyclonal antibodies (RAbs) were obtained. Antibodies against Tb-succinoyl-BSA conjugate (RAb No. 206) were chosen as the best. Under optimized conditions, limit of detection and 50% intercept of our ELISA were 849pg/mL and 8.89ng/mL, respectively. The cross-reactivity (CR) was tested on 10 structurally related compounds and CR did not exceed 6.1%. The reproducibility of the system is expressed as intra- and inter-assay coefficients of variation (9.7% and 11.4%, respectively). Based on conducted experiments, we proposed the use of ELISA for quantification of Tb in complex biological matrices such as plant extracts. A method was applied to analyze three extracts obtained from different parts of L. trilobum. Data obtained were compared to those acquired by UHPLC-MS/MS. The concordance between the methods (103-87%) showed the ability of ELISA to quantify Tb.
- Klíčová slova
- Avidin-biotin, ELISA, Immunoassay, Laser trilobum, Synthesis, Trilobolide,
- MeSH
- Apiaceae chemie MeSH
- butyráty analýza MeSH
- ELISA MeSH
- furany analýza MeSH
- imunoanalýza metody MeSH
- králíci MeSH
- molekulární struktura MeSH
- protilátky imunologie MeSH
- reprodukovatelnost výsledků MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- butyráty MeSH
- furany MeSH
- protilátky MeSH
- trilobolide MeSH Prohlížeč
Rheumatoid arthritis (RA) is a chronic inflammatory disease, leading to progressive destruction of joints and extra-articular tissues, including organs such as liver and spleen. The purpose of this study was to compare the effects of a potential immunomodulator, natural polyphenol N-feruloylserotonin (N-f-5HT), with methotrexate (MTX), the standard in RA therapy, in the chronic phase of adjuvant-induced arthritis (AA) in male Lewis rats. The experiment included healthy controls (CO), arthritic animals (AA), AA given N-f-5HT (AA-N-f-5HT), and AA given MTX (AA-MTX). N-f-5HT did not affect the body weight change and clinical parameters until the 14th experimental day. Its positive effect was rising during the 28-day experiment, indicating a delayed onset of N-f-5HT action. Administration of either N-f-5HT or MTX caused reduction of inflammation measured as the level of CRP in plasma and the activity of LOX in the liver. mRNA transcription of TNF-α and iNOS in the liver was significantly attenuated in both MTX and N-f-5HT treated groups of arthritic rats. Interestingly, in contrast to MTX, N-f-5HT significantly lowered the level of IL-1β in plasma and IL-1β mRNA expression in the liver and spleen of arthritic rats. This speaks for future investigations of N-f-5HT as an agent in the treatment of RA in combination therapy with MTX.
- MeSH
- arachidonátlipoxygenasy genetika metabolismus MeSH
- artritida experimentální farmakoterapie genetika patologie MeSH
- biologické markery MeSH
- C-reaktivní protein MeSH
- časové faktory MeSH
- cytokiny krev genetika metabolismus MeSH
- játra účinky léků metabolismus MeSH
- krysa rodu Rattus MeSH
- mediátory zánětu * MeSH
- methotrexát farmakologie MeSH
- modely nemocí na zvířatech MeSH
- orgánová specificita MeSH
- regulace genové exprese účinky léků MeSH
- serotonin analogy a deriváty farmakologie MeSH
- stupeň závažnosti nemoci MeSH
- transkriptom * MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- arachidonátlipoxygenasy MeSH
- biologické markery MeSH
- C-reaktivní protein MeSH
- cytokiny MeSH
- mediátory zánětu * MeSH
- methotrexát MeSH
- N-feruloylserotonin MeSH Prohlížeč
- serotonin MeSH
Our previous research on immunostimulatory properties of trilobolide and its structurally related natural analogues isolated from Laser trilobum (L.) Borkh., encouraged us to investigate structurally related guaianolides belonging to a specific group of sesquiterpene lactones with characteristic glycol moiety attached to the lactone ring. Ever increasing attention has been paid to certain guaianolides such as thapsigargin and trilobolide for their promising anti-inflammatory, anticancer, anti-infectious and SERCA inhibitory activities. However, due to their alkylation capabilities, they might be cytotoxic. Search for compounds with preserved immunobiological properties and decreased cytotoxicity led us to transform some of their structural features, particularly those related to their side chain functionality. For this reason, we prepared a series of over 20 various deacylated, acyl modified, or relactonized derivatives of trilobolide. The immunobiological effects were screened in vitro using the rat peritoneal cells primed with lipopolysaccharide. Secretion of interferon-γ (IFN-γ), interleukins (IL) IL-1β, IL-6 and tumour necrosis factor-α (TNF-α) were determined by ELISA, and nitric oxide (NO) production by Griess reagent. Relation between the molecular structure and immunobiological activity was investigated. Acetylation at 7-OH and 11-OH positions of the lactone ring, or acyl modification of the guaianolide functionalities (including relactonization) of trilobolide, led to inability to stimulate secretion of cytokines and production of NO. Interestingly, minor structural changes achieved by catalytic hydrogenation or hydrogenolysis retained the original immunoactivity of trilobolide. It can be concluded that several new chemically transformed sesquiterpene lactones resembling the immunobiological properties of trilobolide or thapsigargin were prepared and identified. The implication of the lactone vicinal diol (glycol) moiety, combined with other structure functionality, was confirmed as essential for immune properties of the trilobolide or thapsigargin type of guaianolides.
- Klíčová slova
- Cytokines, Guaianolides, Immunoactivity, Nitric oxide, Sesquiterpene lactones, Trilobolide derivatives,
- MeSH
- Apiaceae chemie MeSH
- butyráty chemie MeSH
- cytokiny metabolismus MeSH
- furany chemie MeSH
- krysa rodu Rattus MeSH
- kultivované buňky MeSH
- laktony chemie farmakologie MeSH
- molekulární struktura MeSH
- oxid dusnatý metabolismus MeSH
- peritoneální makrofágy účinky léků MeSH
- potkani Wistar MeSH
- seskviterpeny chemie farmakologie MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- butyráty MeSH
- cytokiny MeSH
- furany MeSH
- laktony MeSH
- oxid dusnatý MeSH
- seskviterpeny MeSH
- trilobolide MeSH Prohlížeč
- MeSH
- biologické přípravky MeSH
- dějiny 20. století MeSH
- dějiny 21. století MeSH
- kongresy jako téma * dějiny MeSH
- terpeny * MeSH
- Check Tag
- dějiny 20. století MeSH
- dějiny 21. století MeSH
- Publikační typ
- historické články MeSH
- úvodníky MeSH
- Názvy látek
- biologické přípravky MeSH
- terpeny * MeSH
