The goal of this study was to develop novel radioprotective agents targeting the intrinsic apoptotic pathway and thus decreasing the radiation-induced damage. For that purpose, we designed, synthesized and analyzed ten new compounds based on the 1-(4-(2-hydroxyethyl)piperazin-1-yl)-3-phenoxypropan-2-ol leading structure. The cytotoxicity of the newly synthesized substances was tested in vitro on cell lines derived from different progenitor cells by WST-1 proliferation assay. MTT test was utilized to assess half-maximal inhibitory concentrations and maximum tolerated concentrations of novel compounds in A-549 cells. Screening for radioprotective properties was performed using flow-cytometry in MOLT-4 cells exposed to 60Co ionizing gamma radiation. Selected candidates underwent in vivo testing in C57Bl/6 J mice having a positive impact on their immunological status. In summary, we report here promising compounds with radioprotective effect in vivo.
- Klíčová slova
- 1-(2-hydroxyethyl)piperazine derivative, In vitro, Ionizing radiation, Mice, Radioprotection, Synthesis,
- MeSH
- apoptóza účinky léků MeSH
- knihovny malých molekul chemická syntéza chemie farmakologie MeSH
- lidé MeSH
- molekulární struktura MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- proliferace buněk účinky léků MeSH
- propanoly chemická syntéza chemie farmakologie MeSH
- radioprotektivní látky chemická syntéza chemie farmakologie MeSH
- simulace molekulového dockingu MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- knihovny malých molekul MeSH
- propanoly MeSH
- radioprotektivní látky MeSH
The increasing risk of acute large-scale exposure of ionising irradiation on the population underlines the necessity of developing effective radioprotective and mitigating agents. The aim of this work was to investigate the effect of sodium orthovanadate pre-treatment on mice exposed to high doses of gamma rays (from 5 to 13 Gy). The determination of median lethal dose within 30 days confirmed that orthovanadate applied to total-body-irradiated mice intra-peritoneally has a radioprotective but not a mitigating effect. With orthovanadate pre-treatment, the composition of cellularity in the bone marrow improved substantially and the main lymphocyte populations restored during the first month after irradiation. These findings contribute to 'gap-filling' in radioprotective effects and demonstrate the importance of haematological parameters in radiation-response prediction.
- MeSH
- B-lymfocyty účinky léků MeSH
- buňky NK účinky léků MeSH
- celotělové ozáření * MeSH
- inhibitor p21 cyklin-dependentní kinasy metabolismus MeSH
- ionizující záření MeSH
- kostní dřeň účinky léků účinky záření MeSH
- lymfocyty účinky léků účinky záření MeSH
- makrofágy metabolismus MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nádorový supresorový protein p53 metabolismus MeSH
- průtoková cytometrie MeSH
- radioprotektivní látky farmakologie MeSH
- T-lymfocyty účinky léků MeSH
- vanadáty farmakologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- Cdkn1a protein, mouse MeSH Prohlížeč
- inhibitor p21 cyklin-dependentní kinasy MeSH
- nádorový supresorový protein p53 MeSH
- radioprotektivní látky MeSH
- Trp53 protein, mouse MeSH Prohlížeč
- vanadáty MeSH
We report the design, synthesis and biological evaluation of 17 novel 8-aryl-2-morpholino-3,4-dihydroquinazoline derivatives based on the standard model of DNA-PK and PI3K inhibitors. Novel compounds are sub-divided into two series where the second series of five derivatives was designed to have a better solubility profile over the first one. A combination of in vitro and in silico techniques suggested a plausible synergistic effect with doxorubicin of the most potent compound 14d on cell proliferation via DNA-PK and poly(ADP-ribose) polymerase-1 (PARP-1) inhibition, while alone having a negligible effect on cell proliferation.
- Klíčová slova
- Cancer, Chemosensitization, DNA-dependent protein kinase, NU7441, Phosphatidylinositol 3-kinase, Poly(ADP-ribose) polymerase-1,
- MeSH
- apoptóza účinky léků MeSH
- buňky HT-29 MeSH
- chinazolinony chemická syntéza farmakologie toxicita MeSH
- doxorubicin farmakologie MeSH
- inhibitory enzymů chemická syntéza farmakologie toxicita MeSH
- jaderné proteiny antagonisté a inhibitory MeSH
- lidé MeSH
- morfoliny chemická syntéza farmakologie toxicita MeSH
- myši MeSH
- outbrední kmeny zvířat MeSH
- poly(ADP-ribosa)polymerasa 1 antagonisté a inhibitory MeSH
- proliferace buněk účinky léků MeSH
- proteinkinasa aktivovaná DNA antagonisté a inhibitory MeSH
- protinádorové látky farmakologie MeSH
- racionální návrh léčiv MeSH
- synergismus léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- chinazolinony MeSH
- doxorubicin MeSH
- inhibitory enzymů MeSH
- jaderné proteiny MeSH
- morfoliny MeSH
- PARP1 protein, human MeSH Prohlížeč
- poly(ADP-ribosa)polymerasa 1 MeSH
- PRKDC protein, human MeSH Prohlížeč
- proteinkinasa aktivovaná DNA MeSH
- protinádorové látky MeSH
This review summarizes recent progress in understanding the role of p53-upregulated mediator of apoptosis (PUMA) in molecular pathways with respect to its potential therapeutic applications. Particular emphasis is given to the PUMA´s role in ionizing radiation-induced signalling as radiotoxicity of normal tissue is mediated mostly via apoptosis. PUMA and its p53-dependent and p53- independent induction are described and potential use as a new target for the development of radioprotective agents is suggested. Further implications, including targeting PUMA to prevent and treat cardiovascular and neurodegenerative diseases, are also discussed together with an overview of other therapeutic applications. Finally, basic chemical structures for the development of novel PUMA modulators such as pifithrine derivatives, kinase inhibitors or modulators of Bcl-2 protein family are described.
- Klíčová slova
- PUMA, brain, hearth, ionizing radiation, radioprotection, small molecule inhibitor.,
- MeSH
- apoptóza účinky léků genetika MeSH
- cílená molekulární terapie metody MeSH
- kardiovaskulární látky farmakologie terapeutické užití MeSH
- kardiovaskulární nemoci farmakoterapie patologie MeSH
- lidé MeSH
- nádorový supresorový protein p53 antagonisté a inhibitory genetika metabolismus MeSH
- nádory genetika radioterapie MeSH
- neurodegenerativní nemoci farmakoterapie patologie MeSH
- neuroprotektivní látky farmakologie terapeutické užití MeSH
- poškození DNA účinky léků účinky záření MeSH
- proteiny regulující apoptózu antagonisté a inhibitory metabolismus MeSH
- protoonkogenní proteiny antagonisté a inhibitory metabolismus MeSH
- radiační poranění prevence a kontrola MeSH
- radioprotektivní látky farmakologie terapeutické užití MeSH
- signální transdukce účinky léků genetika MeSH
- tolerance záření účinky léků MeSH
- vazba proteinů účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- BBC3 protein, human MeSH Prohlížeč
- kardiovaskulární látky MeSH
- nádorový supresorový protein p53 MeSH
- neuroprotektivní látky MeSH
- proteiny regulující apoptózu MeSH
- protoonkogenní proteiny MeSH
- radioprotektivní látky MeSH
- TP53 protein, human MeSH Prohlížeč
AIM: DNA damage response plays an eminent role in patients' response to conventional chemotherapy and radiotherapy. Its inhibition is of great interest as it can overcome cancer cell resistance and reduce the effective doses of DNA damaging agents. Results & methodology: We have focused our research on phosphatidylinositol 3-kinase-related kinases and prepared 35 novel compounds through a scaffold hopping approach. The newly synthesized inhibitors were tested on a panel of nine cancer and one healthy cell lines alone and in combination with appropriate doses of doxorubicin. CONCLUSION: Five novel compounds 4f, 10b, 15g, 7e and 15f in combination with doxorubicin showed significant antiproliferative effect on seven cancer cell lines while not affecting the cell growth alone.
- Klíčová slova
- ATM kinase *, DNA damage response *, DNA-PK *, LY294002 *, cancer cells chemosensitization *, doxorubicin *,
- MeSH
- doxorubicin farmakologie MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádory farmakoterapie MeSH
- proliferace buněk účinky léků MeSH
- protinádorové látky chemie farmakologie MeSH
- purinony chemie farmakologie MeSH
- pyrimidinony chemie farmakologie MeSH
- pyrroly chemie farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- doxorubicin MeSH
- protinádorové látky MeSH
- purinony MeSH
- pyrimidinony MeSH
- pyrroly MeSH
Scoulerine is an isoquinoline alkaloid, which indicated promising suppression of cancer cells growth. However, the mode of action (MOA) remained unclear. Cytotoxic and antiproliferative properties were determined in this study. Scoulerine reduces the mitochondrial dehydrogenases activity of the evaluated leukemic cells with IC50 values ranging from 2.7 to 6.5 µM. The xCELLigence system revealed that scoulerine exerted potent antiproliferative activity in lung, ovarian and breast carcinoma cell lines. Jurkat and MOLT-4 leukemic cells treated with scoulerine were decreased in proliferation and viability. Scoulerine acted to inhibit proliferation through inducing G2 or M-phase cell cycle arrest, which correlates well with the observed breakdown of the microtubule network, increased Chk1 Ser345, Chk2 Thr68 and mitotic H3 Ser10 phosphorylation. Scoulerine was able to activate apoptosis, as determined by p53 upregulation, increase caspase activity, Annexin V and TUNEL labeling. Results highlight the potent antiproliferative and proapoptotic function of scoulerine in cancer cells caused by its ability to interfere with the microtubule elements of the cytoskeleton, checkpoint kinase signaling and p53 proteins. This is the first study of the mechanism of scoulerine at cellular and molecular level. Scoulerine is a potent antimitotic compound and that it merits further investigation as an anticancer drug.
- MeSH
- aktivace enzymů účinky léků MeSH
- apoptóza účinky léků MeSH
- berberinové alkaloidy chemie farmakologie MeSH
- estery chemie MeSH
- fosforylace účinky léků MeSH
- kaspasy metabolismus MeSH
- kontrolní body buněčného cyklu účinky léků MeSH
- kyseliny karboxylové chemie MeSH
- lidé MeSH
- membránový potenciál mitochondrií účinky léků MeSH
- mikrotubuly účinky léků metabolismus MeSH
- nádorové buněčné linie MeSH
- proliferace buněk účinky léků MeSH
- protinádorové látky chemie farmakologie MeSH
- signální transdukce účinky léků MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zlomy DNA účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- berberinové alkaloidy MeSH
- discretamine MeSH Prohlížeč
- estery MeSH
- kaspasy MeSH
- kyseliny karboxylové MeSH
- protinádorové látky MeSH
Fifteen Amaryllidaceae alkaloids (1-15) of various structural types were isolated by standard chromatographic methods from fresh bulbs of Narcissus poeticus cv. Pink Parasol. The chemical structures were elucidated by MS, and 1D and 2D NMR spectroscopic analyses, and by comparison with literature data. Narcipavline (5) and narcikachnine (6) are reported here for the first time. In their structure are combined two basic structural types of Amaryllidaceae alkaloids (galanthamine- and galanthindole-structural types), which represent a new structural type of these compounds. Alkaloids isolated in sufficient amounts were evaluated for their human erythrocytic acetylcholinesterase, and human serum butyrylcholinesterase (HuBuChE) inhibition activity using Ellman's method. Z-Gly-Pro-p-nitroanilide was used as substrate in the prolyl oligopeptidase (POP) assay. Untested alkaloids were also screened for their cytotoxic activity against a small panel of human cancer cells, which spanned cell lines from different tissue types. In parallel, MRC-5 human fibroblasts were employed to determine overall toxicity against noncancerous cells. Some compounds were evaluated for their antiprotozoal activity. The newly isolated alkaloid narcipavline (5) showed interesting HuBuChE inhibition activity (IC50 = 24.4 ± 1.2 µM), and norlycoramine (11) demonstrated promising POP inhibition (IC50 = 0.21 ± 0.01 mM).
- Klíčová slova
- Alzheimer’s disease, Amaryllidaceae, Antiplasmodial activity, Cytotoxicity, Narcissus poeticus cv. Pink Parasol,
- MeSH
- alkaloidy chemie izolace a purifikace farmakologie MeSH
- buňky A549 MeSH
- buňky HT-29 MeSH
- butyrylcholinesterasa metabolismus MeSH
- cholinesterasové inhibitory chemie izolace a purifikace farmakologie MeSH
- HeLa buňky MeSH
- inhibitory růstu chemie izolace a purifikace farmakologie MeSH
- Jurkat buňky MeSH
- kořeny rostlin MeSH
- lidé MeSH
- MFC-7 buňky MeSH
- myši MeSH
- Narcissus * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- alkaloidy MeSH
- butyrylcholinesterasa MeSH
- cholinesterasové inhibitory MeSH
- inhibitory růstu MeSH
BACKGROUND: Haemanthamine (HA) and sodium butyrate (NaB) are promising candidates for chemotherapy as a treatment for cancer. PURPOSE: We aimed to determine the anticancer potential of HA and NaB, alone and in combination, in A2780 ovarian cancer cells and concurrently investigated anticancer potential in contrast to non-cancer human MRC-5 fibroblasts. METHODS: Antiproliferative effects were determined by WST-1 assay and by Trypan blue exclusion staining. Cell cycle distributions were studied by flow cytometry and protein levels were determined by Western blotting. RESULTS: The combination of HA and NaB caused a significant decrease in the proliferation of A2780 cells compared to the stand-alone treatment of cells by HA or NaB. This effect was less pronounced in non-cancer MRC-5 fibroblasts. In the later intervals, the number of A2780 living cells was strongly decreased by treatment using a combination of NaB and HA. This simultaneous application had no considerable effect in MRC-5 fibroblasts. The combination of NaB and HA led to the suppression of cells in the G1 phase and caused an accumulation of cells in the S and G2 phase in comparison to those treated with NaB and HA alone. Treatment of cells with NaB alone led to the activation of proteins regulating the cell cycle. Notably, p21WAF1/Cip1 was upregulated in both A2780 and MRC-5 cells, while checkpoint kinases 1 and 2 were activated via phosphorylation only in A2780 cells. Unexpectedly, NaB in combination with HA suppressed the phosphorylation of Chk2 on threonine 68 and Chk1 on serine 345 in A2780 cells and downregulated p21WAF1/Cip1 in both tested cell lines. The sensitization of cells to HA and NaB treatment seems to be accompanied by increased histone acetylation. NaB-induced acetylation of histone H3 and H4 and histone acetylation increased markedly when a combination of NaB and HA was applied. Whereas the most prominent hyperacetylation after HA and NaB treatment was observed in A2780 cells, the acetylation of histones occurred in both cell lines. CONCLUSION: In summary, we have demonstrated the enhanced activity of HA and NaB against A2780 cancer cells, while eliciting no such effect in non-cancer MRC-5 cells.
- Klíčová slova
- Cell death, Haemanthamine, Histone acetylation, Ovarian carcinoma cells, Sodium butyrate,
- MeSH
- acetylace MeSH
- aktivace transkripce účinky léků MeSH
- alkaloidy amarylkovitých farmakologie MeSH
- buněčné dělení účinky léků MeSH
- buněčný cyklus účinky léků MeSH
- checkpoint kinasa 1 metabolismus MeSH
- checkpoint kinasa 2 metabolismus MeSH
- fenantridiny farmakologie MeSH
- fosforylace MeSH
- histony metabolismus MeSH
- inhibitor p21 cyklin-dependentní kinasy metabolismus MeSH
- kyselina máselná farmakologie MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádory vaječníků patologie MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- alkaloidy amarylkovitých MeSH
- CDKN1A protein, human MeSH Prohlížeč
- checkpoint kinasa 1 MeSH
- checkpoint kinasa 2 MeSH
- CHEK1 protein, human MeSH Prohlížeč
- CHEK2 protein, human MeSH Prohlížeč
- fenantridiny MeSH
- hemanthamine MeSH Prohlížeč
- histony MeSH
- inhibitor p21 cyklin-dependentní kinasy MeSH
- kyselina máselná MeSH
In this study, twenty-two Amaryllidaceae alkaloids were screened for their anticancer potential. All isolates were evaluated for antiproliferative activities on a panel of 17 human cell types of different tissue origin using WST-1 assay. In addition, we determined the antiproliferative effect with a real-time cell analysis xCELLigence system. Thereafter, to evaluate the barely known in vivo anticancer potential of the most potent molecule haemanthamine, a preliminary study was performed using an Ehrlich tumor-bearing mice model. The results showed that haemanthamine, lycorine and haemanthidine exerted the highest antiproliferative activity. The mean growth percent (GP) value after a single-dose 10 μM treatment was for haemanthamine 21%, for lycorine 21% and for haemanthidine 27% that of untreated control cells (100%). Furthermore, haemanthamine, lycorine and haemanthidine exhibited significant cytotoxicities against all the tested cell lines with individual IC50 values in the micromolar range. Dynamic real-time measures of impedance by xCELLigence indicated that these three compounds suppress cell proliferation after 10 h of treatment at a concentration of 10 μM or higher. Regrettably, in a follow-up in vivo antitumor activity study, haemanthamine showed no statistically significant reduction in the tumor size with no prolongation of survival time of Ehrlich tumor-bearing mice. Taken together, these results provide a new clue and guidance for exploiting Amaryllidaceae alkaloids as anticancer agents.
- Klíčová slova
- Alkaloids, Amaryllidaceae, Antiproliferative activity, In vitro, In vivo,
- MeSH
- alkaloidy chemie farmakologie terapeutické užití MeSH
- Amaryllidaceae chemie metabolismus MeSH
- apoptóza účinky léků MeSH
- Ehrlichův tumor farmakoterapie mortalita patologie MeSH
- fytogenní protinádorové látky chemie farmakologie terapeutické užití MeSH
- Kaplanův-Meierův odhad MeSH
- lidé MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- proliferace buněk účinky léků MeSH
- screeningové testy protinádorových léčiv MeSH
- transplantace heterologní MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- alkaloidy MeSH
- fytogenní protinádorové látky MeSH
BACKGROUND: The search for new anticancer compounds is a crucial element of natural products research. PURPOSE: In this study the effects of naturally occurring homochelidonine in comparison to chelidonine on cell cycle progression and cell death in leukemic T-cells with different p53 status are described. METHODS: The mechanism of cytotoxic, antiproliferative, apoptosis-inducing effects and the effect on expressions of cell cycle regulatory proteins was investigated using XTT assay, Trypan blue exclusion assay, flow cytometry, Western blot analysis, xCELLigence, epi-fluorescence and 3D super resolution microscopy. A549 cells were used for xCELLigence, clonogenic assay and for monitoring microtubule stability. RESULTS: We found that homochelidonine and chelidonine displayed significant cytotoxicity in examined blood cancer cells with the exception of HEL 92.1.7 and U-937 exposed to homochelidonine. Unexpectedly, homochelidonine and chelidonine-induced cytotoxicity was more pronounced in Jurkat cells contrary to MOLT-4 cells. Homochelidonine showed an antiproliferative effect on A549 cells but it was less effective compared to chelidonine. Biphasic dose-depended G1 and G2/M cell cycle arrest along with the population of sub-G1 was found after treatment with homochelidonine in MOLT-4 cells. In variance thereto, an increase in G2/M cells was detected after treatment with homochelidonine in Jurkat cells. Treatment with chelidonine induced cell cycle arrest in the G2/M cell cycle in both MOLT-4 and Jurkat cells. MOLT-4 and Jurkat cells treated with homochelidonine and chelidonine showed features of apoptosis such as phosphatidylserine exposure, a loss of mitochondrial membrane potential and an increase in the caspases -3/7, -8 and -9. Western blots indicate that homochelidonine and chelidonine exposure activates Chk1 and Chk2. Studies conducted with fluorescence microscopy demonstrated that chelidonine and homochelidonine inhibit tubulin polymerization in A549 cells. CONCLUSION: Collectively, the data indicate that chelidonine and homochelidonine are potent inducers of cell death in cancer cell lines, highlighting their potential relevance in leukemic cells.
- Klíčová slova
- Apoptosis, Biphasic cell cycle arrest, Homochelidonine, Mitotic block,
- MeSH
- apoptóza účinky léků MeSH
- benzofenantridiny farmakologie MeSH
- berberinové alkaloidy farmakologie MeSH
- Chelidonium chemie MeSH
- Jurkat buňky MeSH
- kaspasy metabolismus MeSH
- kontrolní body buněčného cyklu účinky léků MeSH
- lidé MeSH
- membránový potenciál mitochondrií účinky léků MeSH
- nádorové buněčné linie účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- benzofenantridiny MeSH
- berberinové alkaloidy MeSH
- chelidonine MeSH Prohlížeč
- homochelidonine MeSH Prohlížeč
- kaspasy MeSH
