RESUMO
The aim of this study was to examine the antitumour effects of plant phenolic acids, gallic acid (GA) and ellagic acid (EA), on human promyelocytic leukaemia sensitive HL60 cell line and its resistant sublines exhibiting two MDR phenotypes: HL60/VINC (overexpressing P-glycoprotein) and HL60/MX2 (characterized by the presence of mutated α isoform of topoisomerase II). Both studied compounds exerted comparable cytotoxic activities towards sensitive HL60 cells and their MDR counterparts. It was also found that GA and EA modulated the cellular level of reactive oxygen species in a dose-dependent and time-dependent manner. Furthermore, it was demonstrated that GA (IC90 ) and EA (IC50 and IC90 ) significantly increased the percentage of sub-G1 subpopulation of all studied leukaemia cells causing oligonucleosomal DNA fragmentation. Both compounds used at IC90 triggered mainly the apoptotic death of these cells. However, GA had no effect on the activity of caspase-3 as well as caspase-8 in sensitive HL60 cells and their MDR counterparts. In contrast, EA provoked a significant activation of these caspases in all studied leukaemia cells. It was also found that lysosomes were not involved in triggering programmed death of sensitive HL60 and MDR cells by GA and EA.
Assuntos
Antineoplásicos/uso terapêutico , Resistência a Múltiplos Medicamentos/efeitos dos fármacos , Ácido Elágico/uso terapêutico , Ácido Gálico/uso terapêutico , Células HL-60/efeitos dos fármacos , Leucemia Promielocítica Aguda/tratamento farmacológico , Polifenóis/uso terapêutico , Antineoplásicos/farmacologia , Ácido Elágico/farmacologia , Ácido Gálico/farmacologia , Humanos , Leucemia Promielocítica Aguda/patologia , Polifenóis/farmacologiaRESUMO
Gongronema latifolium Benth (Asclepiadaceae) is an edible-green-leafy vegetable with known medicinal value. A chemical investigation of the 80% methanolic extract of the leaves led to the isolation of a new pregnane glycoside: iloneoside (3-O-[6-deoxy-3-O-methyl-ß-D-allopyranosyl-(1â14)-ß-D-oleandropyranosyl]-11,12-di-O-tigloyl-17ß-marsdenin), together with four known constituents. Their chemical structures were determined by spectroscopic analysis. The isolates were tested for their in vitro growth inhibitory activity against human leukemia HL-60 cells. Iloneoside was the most active and gave apoptotic response. Molecular docking analysis demonstrated that iloneoside could be accommodated within hot spots of anti-apoptotic protein Bcl-2. These results suggest G. latifolium as a reliable source of potent anticancer compounds.
Assuntos
Antineoplásicos/isolamento & purificação , Apocynaceae/química , Glicosídeos/isolamento & purificação , Folhas de Planta/química , Pregnanos/isolamento & purificação , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Células HL-60/efeitos dos fármacos , Humanos , Simulação de Acoplamento Molecular , Estrutura Molecular , Extratos Vegetais/químicaRESUMO
OBJECTIVE: Mesenchymal stem cells (MSCs) are multipotent stromal cells that can differentiate into a variety of cell types. They control the process of hematopoiesis by secreting regulatory cytokines and growth factors and by the expression of important cell adhesion molecules for cell-to-cell interactions. This investigation was intended to examine the effect of bone marrow (BM)-derived MSCs on the differentiation of HL-60 cells according to morphological evaluation, flow cytometry analysis, and gene expression profile. MATERIALS AND METHODS: The BM-MSCs were cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum (FBS). After the third passage, the BM-MSCs were irradiated at 30 Gy. To compare how the HL-60 cells differentiated in groups treated differently, HL-60 cells were cultured in RPMI-1640 and supplemented with 10% FBS. The HL-60 cells were seeded into six-well culture plates and treated with all-trans-retinoic acid (ATRA), BM-MSCs, or BM-MSCs in combination with ATRA, while one well remained as untreated HL-60 cells. The expression levels of the granulocyte subset-specific genes in the HL-60 cells were assayed by real-time polymerase chain reaction. RESULTS: Our results revealed that BM-MSCs support the granulocytic differentiation of the human promyelocytic leukemia cell line HL-60. CONCLUSION: Based on the results of this study, we concluded that BM-MSCs may be an effective resource in reducing or even preventing ATRA's side effects and may promote differentiation for short medication periods. Though BM-MSCs are effective resources, more complementary studies are necessary to improve this differentiation mechanism in clinical cases.
Assuntos
Comunicação Celular , Diferenciação Celular , Granulócitos/metabolismo , Células HL-60/metabolismo , Células-Tronco Mesenquimais/metabolismo , Biomarcadores , Diferenciação Celular/efeitos dos fármacos , Técnicas de Cocultura , Granulócitos/citologia , Células HL-60/efeitos dos fármacos , Humanos , Imuno-Histoquímica , Imunofenotipagem , Tretinoína/farmacologiaRESUMO
Considering the pressing need for new drugs to treat sleeping sickness and Nagana disease, Mentha crispa essential oil, its principal constituent rotundifolone, and four related p-menthane-type monoterpenes (two stereoisomers of limonene epoxide, perillyl alcohol, and perillyl aldehyde) were investigated for their activity against bloodstream forms of Trypanosoma brucei. The general cytotoxicity of the compounds was determined with human myeloid HL-60 cells. The effect of the M. crispa essential oil and the monoterpenes on the growth of parasite and human cells was evaluated in cell cultures with the resazurin viability assay. Of all of the compounds tested, M. crispa essential oil, rotundifolone, and perillyl aldehyde showed the highest trypanocidal activities with 50â% growth inhibition (GI50) and minimum inhibitory concentration values of 0.3 µg/mL and 1 µg/mL, respectively. In contrast, HL-60 cells were considerably less sensitive to the compounds with minimum inhibitory concentration values of 100 µg/mL and GI50 values ranging between 3.4 to 13.8 µg/mL. As a consequence of this, GI50 and minimum inhibitory concentration ratios of cytotoxic to trypanocidal activity (selectivity index) of these three compounds were promising with values of 11-45 and 100, respectively. These results indicate that the p-menthane-type monoterpenes rotundifolone and perillyl aldehyde are interesting lead candidates for further rational antitrypanosomal drug development.
Assuntos
Mentha/química , Monoterpenos/farmacologia , Óleos Voláteis/farmacologia , Tripanossomicidas/farmacologia , Trypanosoma brucei brucei/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos/métodos , Células HL-60/efeitos dos fármacos , Humanos , Testes de Sensibilidade Microbiana , Óleos Voláteis/química , Estereoisomerismo , Relação Estrutura-Atividade , Tripanossomicidas/químicaRESUMO
BACKGROUND: Abietane diterpenes have attracted much attention because they display a wide range of biological activities, including antitumor activities. These compounds are the most diverse of the diterpenoids isolated from species of Plectranthus. Naturally occurring diterpene parvifloron D is the main phytochemical constituent of Plectranthus ecklonii. To examine the therapeutic potential of the plant, we evaluated whether parvifloron D displays cytotoxicity against human tumor cells. METHODS: The cytotoxicity was analyzed by colorimetric 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay. Apoptosis was evaluated by fluorescent microscopy, transmission electron microscopy, flow cytometric analysis of annexin V-FITC and propidium iodide-stained cells and DNA fragmentation. Protein expression and processing and release of mitochondrial proteins were analyzed by Western blot. Caspase activity was determined using colorimetric substrates. The membrane potential and intracellular reactive oxygen species were detected by flow cytometry. RESULTS: Parvifloron D displays strong cytotoxic properties against leukemia cells (HL-60, U-937, MOLT-3 and K-562) and in particular P-glycoprotein-overexpressing K-562/ADR cells, but has only weak cytotoxic effects on peripheral blood mononuclear cells (PBMCs). Overexpression of the protective mitochondrial proteins Bcl-2 and Bcl-xL did not confer resistance to parvifloron D-induced cytotoxicity. Growth inhibition of HL-60 cells that was triggered by parvifloron D was found to be caused by a rapid induction of apoptotic cell death. This apoptosis was prevented by the non-specific caspase inhibitor z-VAD-fmk, and by the selective caspase-9 inhibitor z-LEHD-fmk. Cell death induced by parvifloron D was found to be (i) associated with the dissipation of the mitochondrial membrane potential and the release of cytochrome c, (ii) amplified by inhibition of extracellular signal-regulated kinases (ERKs) 1/2 signaling and (iii) caused by a mechanism dependent on intracellular reactive oxygen species generation. CONCLUSION: Parvifloron D is a potent cytotoxic compound against several human tumor cells and also a fast and potent apoptotic inducer in leukemia cells.
Assuntos
Abietanos/farmacologia , Apoptose/efeitos dos fármacos , Plectranthus/química , Antineoplásicos Fitogênicos/farmacologia , Caspases/metabolismo , Células HL-60/efeitos dos fármacos , Humanos , Células K562/efeitos dos fármacos , Leucócitos Mononucleares/efeitos dos fármacos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Estrutura Molecular , Espécies Reativas de Oxigênio/metabolismoRESUMO
PURPOSE: The 'two-faced' character of reactive oxygen species (ROS) plays an important role in cancer biology by acting as secondary messengers in intracellular signaling cascades, enhancing cell proliferation and survival, thereby sustaining the oncogenic phenotype. Conversely, enhanced generation of ROS can trigger an oxidative assault leading to a redox imbalance translating into an apoptotic cell death. Intrinsically, cancer cells have higher basal levels of ROS which if supplemented by additional oxidative insult by pro-oxidants can be cytotoxic, an example being Malabaricone-A (MAL-A). MAL-A is a plant derived diarylnonanoid, purified from fruit rind of the plant Myristica malabarica whose anti-cancer activity has been demonstrated in leukemic cell lines, the modality of cell death being apoptosis. This study aimed to compare the degree of effectiveness of MAL-A in leukemic vs. solid tumor cell lines. METHODS: The cytotoxicity of MAL-A was evaluated by the MTS-PMS cell viability assay in leukemic cell lines (MOLT3, K562 and HL-60) and compared with solid tumor cell lines (MCF7, A549 and HepG2); further studies then proceeded with MOLT3 vs. MCF7 and A549. The contribution of redox imbalance in MAL-A induced cytotoxicity was confirmed by pre-incubating cells with an antioxidant, N-acetyl-L-cysteine (NAC) or a thiol depletor, buthionine sulfoximine (BSO). MAL-A induced redox imbalance was quantitated by flow cytometry, by measuring the generation of ROS and levels of non protein thiols using dichlorofluorescein diacetate (CM-H2DCFDA) and 5-chloromethylfluorescein diacetate (CMFDA) respectively. The activities of glutathione peroxidase (GPx), superoxide dismutase, catalase (CAT), NAD(P)H dehydrogenase (quinone 1) NQO1 and glutathione-S-transferase GST were measured spectrophotometrically. The mitochondrial involvement of MAL-A induced cell death was measured by evaluation of cardiolipin peroxidation using 10-N-nonyl acridine orange (NAO), transition pore activity with calcein-AM, while the mitochondrial transmembrane electrochemical gradient (∆ψ(m)) was measured by JC-1, fluorescence being acquired in a flow cytometer. The apoptotic mode of cell death was evaluated by double staining with annexin V-FITC and propidium iodide (PI), cell cycle analysis by flow cytometry and caspase-3 activity spectrophotometrically. The expression of Nrf2 and HO-1 was examined by western blotting. RESULTS: MAL-A demonstrated a higher degree of cytotoxicity in three leukemic cell lines whose IC50 ranged from 12.70 ± 0.10 to 18.10 ± 0.95 µg/ml, whereas in three solid tumor cell lines, the IC50 ranged from 28.10 ± 0.58 to 55.26 ± 5.90 µg/ml. This higher degree of cytotoxicity in MOLT3, a leukemic cell line was due to a higher induction of redox imbalance, evident by both an increased generation of ROS and concomitant depletion of thiols. This was confirmed by pre-incubation with NAC and BSO, wherein NAC decreased MAL-A induced cytotoxicity by 2.04 fold while BSO enhanced MAL-A cytotoxicity and decreased the IC50 by 5.60 fold. However, in solid tumor cell lines (MCF7 and A549), NAC minimally decreased MAL-A induced cytotoxicity, and BSO increased the IC50 by 1.96 and 2.39 fold respectively. Furthermore, the generation of ROS by MAL-A increased maximally in MOLT3 as the fluorescence increased from 44.28 ± 7.85 to 273.99 ± 32.78, and to a lesser degree in solid tumor cell lines, MCF7 (44.28 ± 14.89 to 207.97 ± 70.64) and A549 (37.87 ± 3.24 to 147.12 ± 38.53). In all three cell lines there was a concomitant depletion of thiols as in MOLT3, the GMFC decreased from 340.65 ± 60.39 to 62.67 ± 11.32, in MCF7 (277.82 ± 50.32 to 100.39 ± 31.93) and in A549 (274.05 ± 59.13 to 83.15 ± 21.43). In MOLT3 as compared to MCF7 and A549, decrease in the activities of GPx, CAT, NQO1 and GST was substantially greater. In all cell lines, the MAL-A induced redox imbalance translated into triggering of initial mitochondrial apoptotic events. Here again, MAL-A induced a higher degree of cardiolipin peroxidation in MOLT3 (67.01%) than MCF7 and A549 (29.15% and 44.30%), as also down regulated the mitochondrial transition pore activity from baseline to a higher extent, GMFC being 48.05 ± 2.37 to 10.70 ± 3.97 (MOLT3), 43.55 ± 3.36 to 15.36 ± 0.60 (MCF7) and 39.58 ± 0.4 to 12.65 ± 1.56 (A549). Perturbation of mitochondrial membrane potential evident by a decrease in the ratio of red/green (J-aggregates/monomers) was 134 fold (14.73/0.11) in MOLT3, 45 fold in MCF7 (20.72/0.46) and 34 fold in A549 (22.01/0.64). The extent of apoptosis using a similar concentration of MAL-A was maximal in MOLT3, wherein a 105 fold increase in annexin V binding was evident (0.83 ± 0.51 to 87.08 ± 9.85%) whereas it increased by 43.11 fold in MCF7 (0.69 ± 0.30 to 29.75 ± 11.79%) and 47.52 fold in A549 (0.61 ± 0.31 to 28.99 ± 17.21%). MAL-A induced apoptosis was also associated with a higher degree of caspase-3 activity in MOLT3 vs. MCF7 or A549 which translated into halting of cell cycle progression, evident by an increment in the sub-G0/G1 population [19.26 fold in MOLT3 (0.95 ± 0.45 vs. 18.30 ± 1.90%), 11.01 fold in MCF7 (0.97 ± 0.37 vs. 10.68 ± 0.69%) and 8.58 fold in A549 (1.06 ± 0.45 vs. 9.10 ± 1.05%)]. MAL-A effectively inhibited Nrf2 and HO-1, more prominently in MOLT3. Furthermore, the decreased expression of Nrf2 in MOLT3 correlated with the decreased activities of NQO1 and GST, suggesting that targeting of the Nrf2 anti-oxidant pathway could be considered. CONCLUSION: Taken together, MAL-A a pro-oxidant compound is likely to be more effective in leukemias, meriting further pharmacological consideration.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Resorcinóis/farmacologia , Apoptose/efeitos dos fármacos , Células HL-60/efeitos dos fármacos , Células Hep G2/efeitos dos fármacos , Humanos , Células K562/efeitos dos fármacos , Células MCF-7/efeitos dos fármacos , Myristicaceae/química , OxirreduçãoRESUMO
Quercus suber L. cork contains a diversity of phenolic compounds, mostly low molecular weight phenols. A rising number of reports support with convergent findings that polyphenols evoke pro-apoptotic events in cancerous cells. However, the literature related to the anti-cancer bioactivity of Q. suber L. cork extractives (QSE) is still limited. Herein, we aim to describe the antitumor potential displayed by cork extractives obtained by different extraction methods in the human promyelocytic leukaemia cells. In order to quantify the effects of QSE on cancer cells viability, phosphatidylserine exposure, caspase-3 activity, mitochondrial membrane potential and cell cycle were evaluated. The results indicated that the QSE present a time-dependent and dose-dependent cytotoxicity in the human promyelocytic leukaemia cells. Such a noxious effect leads these leukaemia cells to their death through apoptotic processes by altering the mitochondrial outer membrane potential, activating caspase-3 and externalizing phosphatidylserine. However, cells cycle progression was not affected by the treatments. This study contributes to open a new way to use this natural resource by exploiting its anti-cancer properties. Moreover, it opens new possibilities of application of cork by-products, being more efficient in the sector of cork-based agriculture. Copyright © 2015 John Wiley & Sons, Ltd.
Assuntos
Apoptose/efeitos dos fármacos , Leucemia Mieloide/patologia , Potencial da Membrana Mitocondrial , Extratos Vegetais/farmacologia , Quercus/química , Caspase 3/metabolismo , Ciclo Celular , Células HL-60/efeitos dos fármacos , Humanos , Fenóis/química , Fenóis/farmacologia , Fosfatidilserinas/metabolismo , Extratos Vegetais/químicaRESUMO
BACKGROUND: Natural products are one of the most important sources of drugs used in pharmaceutical therapeutics. Screening of several natural products in the search for novel anticancer agents against human leukemia HL-60 cells led us to identify potent apoptosis-inducing activity in the essential oil fraction from Artemisia capillaris Thunb. flower. METHODS: The cytotoxic effects of extracts were assessed on human leukemia HL-60 cells by XTT assay. Induction of apoptosis was assessed by analysis of DNA fragmentation and nuclear morphological change. The plant name was checked with the plant list website (http://www.theplantlist.org). RESULTS: A purified compound from the essential oil fraction from Artemisia capillaris Thunb. flower that potently inhibited cell growth in human leukemia HL-60 cells was identified as capillin. The cytotoxic effect of capillin in cells was associated with apoptosis. When HL-60 cells were treated with 10(-6) M capillin for 6 h, characteristic features of apoptosis such as DNA fragmentation and nuclear fragmentation were observed. Moreover, activation of c-Jun N-terminal kinase (JNK) was detected after treatment with capillin preceding the appearance of characteristic properties of apoptosis. Release of cytochrome c from mitochondria was also observed in HL-60 cells that had been treated with capillin. CONCLUSION: Capillin induces apoptosis in HL-60 cells via the mitochondrial apoptotic pathway, which might be controlled through JNK signaling. Our results indicate that capillin may be a potentially useful anticancer drug that could enhance therapeutic efficacy.
Assuntos
Apoptose/efeitos dos fármacos , Di-Inos/farmacologia , Mitocôndrias/efeitos dos fármacos , Óleos Voláteis/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Artemisia/química , Citocromos c/metabolismo , Fragmentação do DNA , Flores/química , Células HL-60/efeitos dos fármacos , Humanos , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Óleos de Plantas/farmacologiaRESUMO
Three new brasilane-type sesquiterpenoids, brasilanes A-C (1-3), together with two new alkane derivatives, colisiderin A (4) and 7(E),9(E)-undecandiene-2,4,5-triol (5), were isolated from cultures of the basidiomycete Coltricia sideroides. Their structures were elucidated by NMR and MS data analyses. The absolute configuration of 4 was determined by TDDFT ECD calculations while brasilane-type sesquiterpenoids were isolated from cultures of mushroom for the first time. Compounds 2 and 4 showed weak cytotoxicities against HL-60 and SW480, respectively.
Assuntos
Basidiomycota/química , Sesquiterpenos/química , Agaricales/química , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Linhagem Celular Tumoral/efeitos dos fármacos , Células HL-60/efeitos dos fármacos , Humanos , Estrutura Molecular , Sesquiterpenos/isolamento & purificaçãoRESUMO
Epigallocatechin gallate (EGCG), ellagic acid (EA) and rosmarinic acid (RA) are natural polyphenols exerting cancer chemopreventive effects. Ribonucleotide reductase (RR; EC 1.17.4.1) converts ribonucleoside diphosphates into deoxyribonucleoside diphosphates being essential for DNA replication, which is why the enzyme is considered an excellent target for anticancer therapy. EGCG, EA, and RA dose-dependently inhibited the growth of human HL-60 promyelocytic leukemia cells, exerted strong free radical scavenging potential, and significantly imbalanced nuclear deoxyribonucleoside triphosphate (dNTP) concentrations without distinctly affecting the protein levels of RR subunits (R1, R2, p53R2). Incorporation of (14)C-cytidine into nascent DNA of tumor cells was also significantly lowered, being equivalent to an inhibition of DNA synthesis. Consequently, treatment with EGCG and RA attenuated cells in the G0/G1 phase of the cell cycle, finally resulting in a pronounced induction of apoptosis. Sequential combination of EA and RA with the first-line antileukemic agent arabinofuranosylcytosine (AraC) synergistically potentiated the antiproliferative effect of AraC, whereas EGCG plus AraC yielded additive effects. Taken together, we show for the first time that EGCG, EA, and RA perturbed dNTP levels and inhibited cell proliferation in human HL-60 promyelocytic leukemia cells, with EGCG and RA causing a pronounced induction of apoptosis. Due to these effects and synergism with AraC, these food ingredients deserve further preclinical and in vivo testing as inhibitors of leukemic cell proliferation.
Assuntos
Antineoplásicos/farmacologia , Catequina/análogos & derivados , Cinamatos/farmacologia , Citarabina/farmacologia , Depsídeos/farmacologia , Ácido Elágico/farmacologia , Trifosfato de Adenosina/química , Catequina/farmacologia , Proliferação de Células/efeitos dos fármacos , DNA/biossíntese , Sinergismo Farmacológico , Sequestradores de Radicais Livres/farmacologia , Células HL-60/efeitos dos fármacos , Humanos , Estrutura Molecular , Inibidores da Síntese de Ácido Nucleico/farmacologia , Nucleotídeos de Timina/química , Ácido RosmarínicoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Arrabidaea chica (Bignoniacea) has been used in popular medicine in Brazil to treat inflammation, skin diseases and leukemia. This work aimed to investigate the anti-inflammatory and antitumoral activities of the A. chica aqueous (AE) and ethanol (EE) extracts. MATERIALS AND METHODS: The murine sponge model was used to evaluate the anti-inflammatory and antiangiogenic activities of AE and EE. Accumulation of neutrophil and macrophage in the implants were determined by assaying myeloperoxidase and N-acetyl-glucosaminidase activities and the neovascularization evaluated by the amount of hemoglobin present in the implant using the Drabkin method. The antitumoral activity was evaluated using the MTT colorimetric method against Jurkat, HL60 and MCF-7 cells. Semi-purified fractions F1-F4 from the EE extract were obtained by a liquid-liquid solvent extraction method and their in vitro anti-proliferative effects were also investigated. RESULTS: Ethanol and aqueous extracts of A. chica decreased neutrophil accumulation and hemoglobin content in the sponge implants without altering the level of cytokines (IL-2, IL- 4, IL-5, IFN-γ, TNF-α and VEGF) and the albumin/globulin ratio in the serum of treated animals. There was no sign of toxicity (clinical, laboratory or histopathology). The ethanol extract presented antiproliferative activity (IC50 21.5-36.3 µg/mL) against HL60 and Jurkat cell lineages and proapoptotic activity at 50 µg/mL in HL60 cells. The fraction F1 also demonstrated significant antiproliferative activity (IC50 38.5 µg/mL) and proapoptotic activity against HL60 cells in a dose dependent manner. CONCLUSIONS: Aqueous and ethanol extracts of A. chica attenuate the inflammatory and angiogenic components of the subcutaneous fibrovascular tissue induced by the synthetic matrix in mice. In addition, the ethanol extract from Arrabidaea chica and its fraction F1 presented in vitro antiproliferative activity and could be useful for developing potential chemopreventive substances.
Assuntos
Anti-Inflamatórios/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Bignoniaceae , Neovascularização Patológica/tratamento farmacológico , Extratos Vegetais/farmacologia , Animais , Bignoniaceae/química , Proliferação de Células/efeitos dos fármacos , Citocinas/biossíntese , Células HL-60/efeitos dos fármacos , Humanos , Células Jurkat/efeitos dos fármacos , Células MCF-7/efeitos dos fármacos , Masculino , Camundongos , Folhas de Planta/química , Fator de Necrose Tumoral alfa/biossíntese , Fator A de Crescimento do Endotélio Vascular/biossínteseRESUMO
N-Formyl peptide receptors (FPRs) are G protein-coupled receptors (GPCRs) that play critical roles in inflammatory reactions, and FPR-specific interactions can possibly be used to facilitate the resolution of pathological inflammatory reactions. We here report the synthesis and biological evaluation of six pairs of chiral ureidopropanamido derivatives as potent and selective formyl peptide receptor-2 (FPR2) agonists that were designed starting from our lead agonist (S)-3-(1H-indol-3-yl)-2-[3-(4-methoxyphenyl)ureido]-N-[[1-(5-methoxy-2-pyridinyl)cyclohexyl]methyl]propanamide ((S)-9a). The new compounds were obtained in overall yields considerably higher than (S)-9a. Several of the new compounds showed agonist properties comparable to that of (S)-9a along with higher selectivity over FPR1. Molecular modeling was used to define chiral recognition by FPR2. In vitro metabolic stability of selected compounds was also assessed to obtain preliminary insight on drug-like properties of this class of compounds.
Assuntos
Amidas/química , Avaliação Pré-Clínica de Medicamentos/métodos , Receptores de Formil Peptídeo/agonistas , Receptores de Lipoxinas/agonistas , Amidas/síntese química , Animais , Cálcio/metabolismo , Técnicas de Química Sintética , Estabilidade de Medicamentos , Células HL-60/efeitos dos fármacos , Humanos , Camundongos Endogâmicos BALB C , Microssomos Hepáticos/efeitos dos fármacos , Ativação de Neutrófilo/efeitos dos fármacos , Ratos , Receptores de Formil Peptídeo/química , Receptores de Lipoxinas/química , Especificidade da Espécie , EstereoisomerismoRESUMO
The 20R- and 20S-isomers of 25-hydroxy-2-methylene-vitamin D3 and 3-desoxy-1α,25-dihydroxy-2-methylene-vitamin D3 have been synthesized. Two alternative synthetic routes were devised for preparation of the required A-ring synthons, starting from the chiral compound derived from the (-)-quinic acid and, alternatively, from the commercially available achiral precursor, monoprotected 1,4-cyclohexanedione. The A-ring dienynes were coupled by the Sonogashira process with the respective C,D-ring fragments, the enol triflates derived from the protected (20R)- or (20S)-25-hydroxy Grundmann ketones. All four compounds possessed significant in vivo activity on bone calcium mobilization and intestinal calcium transport. The presence of a 2-methylene group increased intestinal calcium transport activity of all four analogues above that of the native hormone, 1α,25-dihydroxyvitamin D3. In contrast, bone calcium mobilization was equal to that produced by 1α,25-dihydroxyvitamin D3 in compounds having a (20S)-configuration or diminished to one-tenth that of 1α,25-dihydroxyvitamin D3 in compounds with a (20R)-configuration.
Assuntos
Calcifediol/química , Cálcio/metabolismo , 25-Hidroxivitamina D3 1-alfa-Hidroxilase/metabolismo , Animais , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/metabolismo , Calcifediol/farmacologia , Calcitriol/análogos & derivados , Calcitriol/química , Técnicas de Química Sintética , Avaliação Pré-Clínica de Medicamentos/métodos , Células HL-60/efeitos dos fármacos , Humanos , Mucosa Intestinal/metabolismo , Intestinos/efeitos dos fármacos , Masculino , Estrutura Molecular , Ratos Sprague-Dawley , Receptores de Calcitriol/metabolismo , Relação Estrutura-AtividadeRESUMO
Trypanosoma brucei is the causative agent of sleeping sickness, a fatal disease prevalent in sub-Saharan Africa. The few currently available drug treatments are dated and face problems with toxicity and resistance. For these reasons, there is an urgent need for the development of new chemotherapies for the treatment of sleeping sickness. In this study, we investigated the trypanocidal activity of bitter melon extract. Recently, it has been shown that bitter melon extracts display cytotoxic activity towards different cancer cell lines. However, agents exhibiting anti-tumour activity are usually also inhibiting the growth of T. brucei. Treatment of bloodstream forms of T. brucei with extracts prepared from Chinese and Indian bitter melon varieties resulted in a decrease in cell proliferation. In contrast, human myeloid leukaemia HL-60 cells were 3-6 times less sensitive to the extracts than trypanosomes. Initial fractionation of bitter melon extracts indicated that the trypanocidal activity of the extract is associated with at least two different classes of substances: one class of larger molecular weight compounds (>3 kDa) causing rapid lysis of trypanosomes and one class of smaller molecular weight compounds (<3 kDa) inducing accumulation of the parasites in the G(2)-M phase of the cell cycle. Together, the results suggest that bitter melon is a promising source for trypanocidal agents which could be used as lead compounds for the development of novel anti-sleeping sickness drugs.
Assuntos
Momordica charantia/química , Extratos Vegetais/farmacologia , Tripanossomicidas/farmacologia , Trypanosoma brucei brucei/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Citometria de Fluxo , Células HL-60/efeitos dos fármacos , Humanos , Testes de Sensibilidade Parasitária , Extratos Vegetais/toxicidade , Tripanossomicidas/toxicidade , Trypanosoma brucei brucei/crescimento & desenvolvimentoRESUMO
Fatty acid synthase (FAS) has been proposed to be a new drug target for the development of anticancer agents because of the significant difference in expression of FAS between normal and tumour cells. Since a n-hexane-soluble extract from Ginkgo biloba was demonstrated to inhibit FAS activity in our preliminary test, we isolated active compounds from the n-hexane-soluble extract and evaluated their cytotoxic activity in human cancer cells. Three ginkgolic acids 1-3 isolated from the n-hexane-soluble extract inhibited the enzyme with IC(50) values 17.1, 9.2 and 10.5 µM, respectively, and they showed cytotoxic activity against MCF-7 (human breast adenocarcinoma), A549 (human lung adenocarcinoma) and HL-60 (human leukaemia) cells. Our findings suggest that alkylphenol derivatives might be a new type of FAS inhibitor with cytotoxic activity.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Inibidores Enzimáticos/farmacologia , Ácido Graxo Sintases/antagonistas & inibidores , Ginkgo biloba/química , Salicilatos/farmacologia , Adenocarcinoma/tratamento farmacológico , Adenocarcinoma/patologia , Adenocarcinoma de Pulmão , Antineoplásicos Fitogênicos/química , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Ensaios de Seleção de Medicamentos Antitumorais , Inibidores Enzimáticos/química , Feminino , Células HL-60/efeitos dos fármacos , Hexanos/química , Humanos , Concentração Inibidora 50 , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/patologia , Estrutura Molecular , Extratos Vegetais/química , Folhas de Planta/química , Salicilatos/química , Salicilatos/isolamento & purificaçãoRESUMO
BACKGROUND: We recently reported that the MeOH extract from bulbs of Odontioda Marie Noel 'Velano' exhibited diverse biological activities but most of the activity was concentrated into the EtOAc layer separated by sequential organic solvent extractions. In the present study, the EtOAc layer was subjected to silica-gel column chromatography for further separation into five fractions, and the cytotoxicity and apoptosis-inducing activity of the fractions against human normal oral and tumor cells was further investigated. MATERIALS AND METHODS: Cytotoxic activity was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. The 50% cytotoxic concentration (CC(50)) was determined by the dose-response curve. Tumor specificity (TS) was determined by the ratio of the mean CC(50) for normal cells to the one of tumor cell lines. DNA fragmentation was assayed by agarose gel electrophoresis, caspase-3/-7 activation was monitored by cleavage of substrates either spectrophotometrically or by western blot analysis. RESULTS: Among five fractions, the most hydrophobic fraction (Fr. 1) showed the highest cytotoxicity against all cell lines tested, followed by Fr. 2 >Fr. 3 >Fr. 4 >Fr. 5, in order of increasing polarity. Fr. 2 had the highest tumor-specificity, followed by Fr. 3, Fr. 4, Fr. 1 and Fr. 5. Fr. 1 induced caspase-3 activation more potently in promyelocytic leukemia HL-60 cells, than in oral squamous cell carcinoma (OSCC) HSC-2 cells, whereas it did not induce internucleosomal DNA fragmentation in either of these cell lines. CONCLUSION: The present study suggests that hydrophobic substances in the EtOAc extract of Odontioda Marie Noel 'Velano' exhibit tumor-specific cytotoxicity without inducing apoptosis in the HSC-2 OSCC cell line.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Carcinoma de Células Escamosas/patologia , Neoplasias Bucais/patologia , Orchidaceae/química , Extratos Vegetais/farmacologia , Acetatos , Antineoplásicos Fitogênicos/isolamento & purificação , Caspase 3/metabolismo , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral/efeitos dos fármacos , Cromatografia em Gel , Fragmentação do DNA/efeitos dos fármacos , DNA de Neoplasias/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Ativação Enzimática/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Células HL-60/efeitos dos fármacos , Células HL-60/enzimologia , Humanos , Interações Hidrofóbicas e Hidrofílicas , Raízes de Plantas/química , SolventesRESUMO
BACKGROUND: We recently reported that the MeOH extract of aerial parts and root of Rhinacanthus nasutus showed diverse biological activity, with most activity being concentrated into the EtOAc layer separated by sequential organic solvent extractions. In the present study, the EtOAc extracts were further separated by silica-gel column chromatography into five fractions (Frs. 1-5), and their cytotoxicity and apoptosis-inducing activity investigated. MATERIALS AND METHODS: Cytotoxic activity was determined by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) method. The 50% cytotoxic concentration (CC(50)) was determined from the dose-response curve. Tumor specificity (TS) was determined by the ratio of the mean CC(50) for normal cells to the one for tumor cell lines. DNA fragmentation was assayed by agarose gel electrophoresis. Caspase-3/-7 activation was monitored by cleavage of substrates either spectrophotometrically or by western blot analysis. RESULTS: Among five fractions of the EtOAc extract, Fr. 1, eluted with CHCl(3)-MeOH (50:1), showed the highest tumor specificity (TS=3.3) as compared with other fractions eluted at higher concentrations of MeOH in CHCl(3) (TS=1.0-2.8). Fr. 1 did not induce internucleosomal DNA fragmentation or induced only marginal level of caspase-3 activity in either human promyelocytic leukemia HL-60 cells and human oral squamous cell carcinoma (OSCC) cell lines HSC-2. CONCLUSION: The present study suggests that hydrophobic substances of EtOAc extract show tumor specific cytotoxicity by inducing little or no apoptosis.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Carcinoma de Células Escamosas/patologia , Magnoliopsida/química , Neoplasias Bucais/patologia , Extratos Vegetais/farmacologia , Acetatos , Antineoplásicos Fitogênicos/isolamento & purificação , Caspase 3/metabolismo , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral/efeitos dos fármacos , Cromatografia em Gel , Fragmentação do DNA/efeitos dos fármacos , DNA de Neoplasias/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Ativação Enzimática/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Células HL-60/efeitos dos fármacos , Células HL-60/enzimologia , Humanos , Interações Hidrofóbicas e Hidrofílicas , Raízes de Plantas/química , Caules de Planta/química , SolventesRESUMO
Through bioassay-guided fractionation, the EtOAc extract of a culture broth of the endophytic fungus Phoma species ZJWCF006 in Arisaema erubescens afforded a new α-tetralone derivative, (3S)-3,6,7-trihydroxy-α-tetralone (1), together with cercosporamide (2), ß-sitosterol (3), and trichodermin (4). The structures of compounds were established on the basis of spectroscopic analyses. Compounds 1, 2, and 3 were obtained from Phoma species for the first time. Additionally, the compounds were subjected to bioactivity assays, including antimicrobial activity, against four plant pathogenic fungi (Fusarium oxysporium, Rhizoctonia solani, Colletotrichum gloeosporioides, and Magnaporthe oryzae) and two plant pathogenic bacteria (Xanthomonas campestris and Xanthomonas oryzae), as well as in vitro antitumor activities against HT-29, SMMC-772, MCF-7, HL-60, MGC80-3, and P388 cell lines. Compound 1 showed growth inhibition against F. oxysporium and R. solani with EC50 values of 413.22 and 48.5 µg/mL, respectively. Additionally, compound 1 showed no cytotoxicity, whereas compound 2 exhibited cytotoxic activity against the six tumor cell lines tested, with IC50 values of 9.3 ± 2.8, 27.87 ± 1.78, 48.79 ± 2.56, 37.57 ± 1.65, 27.83 ± 0.48, and 30.37 ± 0.28 µM, respectively. We conclude that endophytic Phoma are promising sources of natural bioactive and novel metabolites.
Assuntos
Antibacterianos/metabolismo , Antifúngicos/metabolismo , Antineoplásicos/metabolismo , Arisaema/microbiologia , Ascomicetos/metabolismo , Endófitos/metabolismo , Antibacterianos/química , Antibacterianos/farmacologia , Antifúngicos/química , Antifúngicos/farmacologia , Antineoplásicos/química , Antineoplásicos/farmacologia , Ascomicetos/crescimento & desenvolvimento , Ascomicetos/isolamento & purificação , Benzofuranos/química , Benzofuranos/metabolismo , Benzofuranos/farmacologia , Linhagem Celular Tumoral/efeitos dos fármacos , Meios de Cultivo Condicionados/química , Endófitos/crescimento & desenvolvimento , Endófitos/isolamento & purificação , Fungos/efeitos dos fármacos , Células HL-60/efeitos dos fármacos , Células HT29/efeitos dos fármacos , Humanos , Medicina Tradicional Chinesa , Doenças das Plantas/microbiologia , Sitosteroides/química , Sitosteroides/metabolismo , Sitosteroides/farmacologia , Especificidade da Espécie , Tetralonas/química , Tetralonas/metabolismo , Tetralonas/farmacologia , Tricodermina/química , Tricodermina/metabolismo , Tricodermina/farmacologia , Xanthomonas/efeitos dos fármacosRESUMO
Scutellaria is a genus of Lamiaceae with known antiproliferative potentials. Scutellaria litwinowii Bornm. & Sint. ex Bornm. is one of the Iranian species of Scutellaria. Although there are widespread reports about the cytotoxic and antitumor effects of some species of this genus, research on the molecular mechanism responsible for the anticancer effects of S. litwinowii has not yet been conducted. In the current study, the apoptotic effects of S. litwinowii on 2 myeloid cell lines, apoptosis-proficient HL60 cells and apoptosis-resistant K562 cells, were analyzed. An increase in the activity of caspases-3, -8, and -9, poly (ADP ribose) polymerase (PARP) cleavage, detection of phosphatidylserine on the outer layer of cell membrane and sub-G1 peak in the flow cytometry histogram of treated cells, suggested the induction of apoptosis. S. litwinowii also increased the Bax/Bcl-2 ratio. It could be concluded that S. litwinowii induced apoptosis in both apoptosis-proficient and apoptosis-resistant leukemic cells and it might be considered as a novel candidate in the treatment of hematological malignancies.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Extratos Vegetais/farmacologia , Scutellaria/química , Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Caspase 8/metabolismo , Caspase 9/metabolismo , Ensaios de Seleção de Medicamentos Antitumorais , Células HL-60/efeitos dos fármacos , Humanos , Células K562/efeitos dos fármacos , Leucemia Promielocítica Aguda/tratamento farmacológico , Leucemia Promielocítica Aguda/patologia , Fosfatidilserinas/metabolismo , Extratos Vegetais/química , Poli(ADP-Ribose) Polimerases/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteína X Associada a bcl-2/metabolismoRESUMO
The objective of the present work was to evaluate the biological activities of the major bioactive compound, xanthatin, and other compounds from Xanthium strumarium (Asteraceae) leaves. Inhibition of bloodstream forms of Trypanosoma brucei brucei and leukaemia HL-60 cell proliferation was assessed using resazurin as a vital stain. Xanthatin was found to be the major and most active compound against T. b. brucei with an IC(50) value of 2.63 µg/mL and a selectivity index of 20. The possible mode of action of xanthatin was further evaluated. Xanthatin showed antiinflammatory activity by inhibiting both PGE(2) synthesis (24% inhibition) and 5-lipoxygenase activity (92% inhibition) at concentrations of 100 µg/mL and 97 µg/mL, respectively. Xanthatin exhibited weak irreversible inhibition of parasite specific trypanothione reductase. Unlike xanthatin, diminazene aceturate and ethidium bromide showed strong DNA intercalation with IC(50) values of 26.04 µg/mL and 44.70 µg/mL, respectively. Substantial induction of caspase 3/7 activity in MIA PaCa-2 cells was observed after 6 h of treatment with 100 µg/mL of xanthatin. All these data taken together suggest that xanthatin exerts its biological activity by inducing apoptosis and inhibiting both PGE(2) synthesis and 5-lipoxygenase activity thereby avoiding unwanted inflammation commonly observed in diseases such as trypanosomiasis.