Human Uterus Carcinoma Research Articles

Medicinal Importance of Flavonoid “Eupatorin” in the Health Sectors: Therapeutic Benefit and Pharmacological Activities Through Scientific Data Analysis

Background: Herbal medicines have been used in the health sector for the treatment of human disorders from the ancient time till the modern age. Plant based herbal products have an important place in medicine due to their health beneficial potential and pharmacological activities. Traditional medicines have been used in developing country, such as India to the developed countries such as the USA and Europe. Traditional medicines have been used in the health sectors for the treatment of numerous types of human disorders. Phytoconstituents play an important role in the growth, reproduction and development of the human being. Plant products have been used for the treatment of skin disorders, wounds, and various forms of inflammatory diseases. Flavonoids are a colorful component of fruits, seeds, vegetables, nuts and plants. Flavonoids have been used for the treatment of human disorders due to their vast pharmacological activities. Methods: Scientific information on eupatorin has been collected from different literature sources and analyzed in order to know the health benefit of eupatorin in medicine. All the scientific information presented here is collected from various databases such as Google, PubMed, and Science Direct etc. Pharmacological activity data of eupatorin have been collected from various literature databases and analyzed here to know the beneficial health potential of eupatorin in medicine. However, important analytical tools for the separation, isolation and identification of eupatorin have also been discussed in the present paper. Results: Eupatorin (39,5-dihydroxy-49,6,7-trimethoxyflavone) is a flavone class flavonoidal compound found to be present in the Orthosiphon stamineus, Salvia mirzayanii, Lantana montevidensis, Lantana montevidensis and Tanacetum vulgare. Eupatorin have been used against bronchitis, stomach disorders and various forms of cancer. Eupatorin have a potential role in various cancerous disorders, such as human uterus carcinoma, human gastric adenocarcinoma, breast carcinoma, melanoma and colon carcinoma. Pharmacological data analysis revealed the effectiveness of eupatorin in human cancer and various types of inflammatory disorders. However, the effectiveness of eupatorin against microorganism and enzymes were also presented in the present work with their herbicidal and vasorelaxation potential in the medicine. Eupatorin has a valuable role in the development of adjuvant chemotherapeutic agents. Conclusion: Present review will be beneficial to the researcher to understand the physiological importance of eupatorin in medicine for the development of newer and effective medicine.

Antiproliferative and Antioxidant Activities of Two Extracts of the Plant Species Euphorbia dendroides L.

Background: These days, the desire for naturally occurring antioxidants has significantly increased, especially for use in foodstuffs, cosmetics, and pharmaceutical products, to replace synthetic antioxidants that are regularly constrained due to their carcinogenicity. Methods: The study in hand aimed to appraise the antioxidant effect of two Euphorbia dendroides extracts using reducing power, anti-peroxidation, and DPPH (1,1 Diphenyl 2 Pycril Hydrazil) scavenging essays, in addition to the anticancer activity against two tumor cell lines, namely C6 (rat brain tumor)cells, and Hela (human uterus carcinoma)cell lines. Results: The results indicated that the ethyl acetate extract exhibited antiradical activity of 29.49%, higher than that of n-butanol extract (18.06%) at 100 µg/mL but much lower than that of gallic acid (78.21%).The ethyl acetate extract exhibits better reducing capacity and lipid peroxidation inhibitory activity compared to n-butanol extract but less than all tested standards. Moreover, the ethyl acetate extract was found to have an antiproliferative activity of more than 5-FU (5-fluoro-Uracil) against C6 cells at 250 µg/mL with IC50 and IC75 of 113.97, 119.49 µg/mL, respectively, and good cytotoxic activity against the Hela cell lines at the same concentration. The HPLC-TOF-MS (high performance liquid chromatography-Time-of-flight-Mass Spectrometry) analyses exposed the presence of various compounds, among which Gallic and Chlorogenic acids functioned as major compounds. Conclusions: The two extracts exhibited moderate anticancer abilities and behaved somewhat as average antioxidant agents. Based on the total phenolics and flavonoids contents, as well as HPLC results, it could be concluded that antiproliferative and antioxidant activities depend upon the content of different phenolics and flavonoids.

Phytochemicals of Chrysophthalmum montanum (DC.) Boiss. Roots and Their Antiproliferative Activities Against HeLa and C6 Cell Lines

Pre-fractionation and contemporary spectroscopic analysis provides important clues with regard to synergistic and antagonistic effects as well as biological activities in complex multi-mixtures, especially natural products. Phytochemical contents of the Chrysophthalmum montanum root extracts were analyzed by gas chromatography–mass spectrometry and high performance liquid chromatography–time of flight/mass spectrometry. Furthermore, biological activity of root extract was investigated by antiproliferative efficiencies against human uterus carcinoma and rat brain tumor cells in vitro. The biological activity tests were carried out as dose-dependent assay using a multiwell spectrophotometer. 5-Fluorouracil was used as standard anticancer drug. Total 61 lipophilic components were identified quantitatively from the sub-fraction of n-hexane in which saturated fatty acids (33.06%), unsaturated fatty acids (21.03%) and other organic components (43.35%). Twenty three hydrophilic components, 16 phenolic and 7 flavonoid compounds, were quantified in the hydrophilic sub-fractions obtained from the C. montanum roots. Among the hydrophilic components, the vanillic acid was analyzed as 46.463 ± 0.167 mg compound/g dry extract in the sub-fraction dichloromethane. The linoleic acid was analyzed as a substantial component (8.13%) in the sub-fraction n-hexane. The highest antiproliferative activities were obtained from the sub-fraction dichloromethane against HeLa cells (IC50 = 36.53 μg/mL) and the sub-fraction n-hexane against C6 cells (IC50 = 11.98 μg/mL). No cytotoxic effect was observed from these sub-fractions.

Bioactivity–guided isolation of flavonoids from Cynanchum acutum L. subsp. sibiricum (willd.) Rech. f. and investigation of their antiproliferative activity

Cynanchum acutum L. subsp. sibiricum (Willd.) Rech. f. was extracted with hexane, acetone, methanol and water individually. A sample was heated in water then extracted with ethyl acetate. Among the extracts, the ethyl acetate extract exhibited the most antiproliferative activity, so isolation of bioactive compounds was carried out from this extract. A new compound, kaempferol-3-O-β-xylopyranosyl-(1-2)-β-rhamnopyranoside (1) along with five known compounds, quercetin-3-O-β-xyloside (2), kaempferol-3-O-β-glucoside (3), quercetin-3-O-β-glucoside (4), kaempferol-3-O-β-rhamnopyranoside (5), and kaempferol-3-O-β-d-neohesperidoside (6) were isolated from ethyl acetate extract. The structures were elucidated by spectroscopic techniques, basically 1D NMR, 2D NMR and LC-TOF/MS. Antiproliferative effects of isolated compounds were determined by xCELLigence using the HeLa (human uterus carcinoma) cell lines. Compound 2 and compound 5 revealed the good antiproliferative activity against HeLa cell lines.

Synthesis and in vitro anticancer evaluation of 1,4-phenylene-bis-pyrimidine-2-amine derivatives

A series of 1,4-phenylene-bis-chalcones 3a-3h were synthesized by the reaction of terephthalaldehyde with substituted arylketones in this study. The novel 1,4-phenylene-bis-pyrimidine-2-amine derivatives 5a-5h were obtained by the addition of guanidine hydrochloride to 1,4-phenylene-bis-chalcone 3a-3h in ethanolic KOH under reflux conditions. The structure of the compounds was explained by means of IR, $^{1}$H NMR, $^{13}$C NMR, and elemental analyses. The anticancer activities of 3a-3h and 5a-5h were investigated against rat brain tumor cells and human uterus carcinoma in vitro$.$ Activity tests were performed as dose-dependent assays at eight concentrations. The positive control was 5-fluorouracil (5-FU). Compounds 3c and 3d were examined and they showed high activities as compared to 5-FU against C6 (rat brain tumor) and HeLa (human uterus carcinoma) cells. The anticancer activity of 5h was better than that of 5-FU at high concentrations cell-selectively against C6 cells.

Synthesis and anticancer and cytotoxic effects of novel 1,4-phenylene-bis-N-thiocarbamoylpyrazole and 1,4-phenylene-bis-pyrazolylthiazole derivatives

Thiazolylpyrazoline derivatives were recently reported as potent anticancer agents. In this study, novel 1,4-phenylene-bis-N-thiocarbamoylpyrazoles (3a-h and 1,4-phenylene-bis-pyrazolylthiazoles (5a-h were prepared and screened for their anticancer activities against C6 (rat brain tumor cells) and HeLa (human uterus carcinoma). Anticancer activity studies were performed as a dose-dependent assay starting with eight concentrations. 5-Fluorouracil (5-FU) was used as a positive control. Compounds 3c, 3d, and 3h were examined and they revealed almost the same activities compared with 5-FU in terms of cell selectivity against C6 cells. Moreover, compounds 3a-h had lower cytotoxicity than 5-FU. The low cytotoxicity values of 3a-h as well as their high antiproliferative activity were encouraging, but further studies are required on the use of these molecules as anticancer drugs.

Synthesis, structural characterization, and antiproliferative/cytotoxic effects of a novel modified poly(maleic anhydride-co-vinyl acetate)/doxorubicin conjugate

Drug carrier, poly(maleic anhydride-co-vinyl acetate) (MAVA or poly[MA-co-VA]) copolymer, was traditionally synthesized by free radical chain polymerization reaction, in methyl ethyl ketone (MEK) organic media at 80 °C, using benzoyl peroxide (BPO) as the radicalic initiator. The purified copolymer was then modified with a chemotherapeutic agent, doxorubicin hydrochloride (DOX) at 75 °C for 72 h, using N-(3-dimethyl-aminopropyl)-N′-ethylcarbodiimide hydrochloride (EDAC) as the carboxylic acid-activating agent. Structural characterization of the MAVA and the modified MAVA/DOX conjugate was carried out by Fourier transform infrared (FTIR) and nuclear magnetic resonance (1H-NMR and 13C-NMR). Their molecular weights were determined by size-exclusion chromatography (SEC). The spectroscopic and SEC results confirmed that conjugated/modification reaction was successfully carried out. UV spectrophotometric measurements indicated that MAVA/DOX preserved its molecular stability in physiological body fluid, PBS (physiological pH 7.40 at 37 °C). Antiproliferative activities of MAVA/DOX were determined by BrdU cell proliferation ELISA assay using C6 (Rat Brain tumor cells) and HeLa (human uterus carcinoma) cell lines in vitro by comparing with free DOX agent (reference compound). Although MAVA showed low antiproliferative activity, both MAVA/DOX and DOX exhibited greater activity against HeLa and C6. Lactate dehydrogenase (LDH) leakage assay was performed for MAVA/DOX and DOX, which detected a non-toxic effect against C6 even at the highest dose (100 μg/mL). IC50 and IC75 values were also determined using ED50 plus v1.0. Molecular modeling at M06-L/6-31 + G(d,p)//AM1 level showed that the electron density in MAVA/DOX is more localized resulting a higher polarization and thereby a higher dipole moment which shed light on the solubility of MAVA/DOX conjugate.

Synthesis and anticancer activities of 1,4-phenylene-bis-N-acetyl- and N-phenylpyrazoline derivatives

Novel 1,4-phenylene-bis-N-acetyl- (3a–h) and bis-N-phenylpyrazoline derivatives (4a–h) were obtained by addition of hydrazine hydrate and phenylhydrazine to bis-chalcone derivatives (1a–h) in acetic acid and acetic acid/ethanol for 4 and 8 h in reflux conditions, respectively. The structures of the obtained bis-N-acetylpyrazoline and bis-N-phenylpyrazoline derivatives were characterized by nuclear magnetic resonance (NMR) and infrared (IR) spectroscopic methods and elemental analysis. Compounds 3a–h and 4a–h were investigated to evaluate their anticancer activities against C6 (rat brain tumor cells) and HeLa (human uterus carcinoma) in vitro using a dose-dependent assay from 5 to 100 μM with 5-fluorouracil (5-FU) as standard anticancer drug. Compound 3a showed higher cell-selective activity compared with 5-FU against HeLa cells. Compounds 3a–h (except 3d) were shown to have better activities than 5-FU against both cells, particularly at high concentration. Compound 4c showed higher cell-selective activity compared with 5-FU against C6 cells. Compound 3a may be particularly promising as an anticancer drug against HeLa cells.

Studies on the Antioxidant and Antiproliferative Potentials ofCirsium arvensesubsp.vestitum

The antioxidant (DPPH, superoxide anion radical scavenging and FRAP) and antiproliferative activities against Vero cells (african green monkey kidney), C6 cells (rat brain tumor cells) and HeLa cells (human uterus carcinoma) of Cirsium arvense (CA) of all parts [root (Kinghorn et al. 2004), stem (CAS), flower (CAF) and leaves] and isolated compounds were investigated in vitro. CAR was shown to have the highest antiproliferative activity against HeLa and C6 cell lines as compared to other parts. A flavon glucoside (linarin), a steroid glucoside (daucosterin), a steroid (stigmasterol) and a new benzymidazole molecule (E−6-(3-(5,6-dimethoxy-1H-benzo[d] imidazol-2-yl)allyloxy)−5-(hydroxymethyl)-tetrahydro-2H-pyran-2,3,4-triol) were isolated from the CAR. The structures of these molecules were determined by the 13C (carbon) and 1H (proton)- Nuclear Magnetic Resonance (NMR), Fourier Transform Infrared Spectroscopy (FT-IR), and Gas Chromatography-Mass Spectrometry (GC-MS) techniques. CAR for C6 cell line, CAS for HeLa and Vero cell line were observed the highest antiproliferative activity. In addition, CAR showed the stronger DPPH activity than α-tocopherol and BHT at high concentrations. The FRAP and superoxide anion radical scavenging activities of CAR (Fr-7) were found to be higher than BHT. Practical Applications Cirsium species are considered as agricultural weed among villagers in Turkey and are used as alternative medicinal plant for years. Studies have shown that these species are rich with regard to bioactive seconder metabolites. There is no phytochemical/bioactivity study about this plant until now. Our findings showed that the extracts of this plant possessed potent antioxidant activities. Four compounds, one of which is a new one to literature, were isolated and characterized. Finally, Cirsium arvense, being considered as an invader plant in agricultural fields, can be evaluated as a good source of antioxidant products.

Chemical constituents and antiproliferative effects of cultured Mougeotia nummuloides and Spirulina major against cancerous cell lines.

In this study, the effect of Mougeotia nummuloides and Spirulina major on Vero cells (African green monkey kidney), C6 cells (rat brain tumor cells) and HeLa cells (human uterus carcinoma) was investigated in vitro. The antiproliferative effect of the methanol extract of M. nummuloides and S. major compared with 5-fluorourasil (5-FU) and cisplatin was tested at various concentrations using the BrdU Cell Proliferation ELISA. Both M. nummuloides and S. major extracts significantly inhibited the proliferation of Vero, HeLa and C6 cancer cell lines with IC50 and IC75 values. The M. nummuloides extract exhibited higher activity than 5-FU and cisplatin on Vero and C6 cells at high concentrations. The S. major extract revealed better antifproliferative activity than standards against Vero cells at 500 μg/mL. The compounds of methanol extracts were determined by GC-MS after the silylation process. Trehalose, monostearin and 1-monopalmitin were detected as major products in the M. nummuloides extract where as in the S. major extract; monostearin, 1-monopalmitin and hexyl alcohol were the main constituents.

New bio-active azo-azomethine based Cu(II) complexes

In this study, new azo-azomethine ligands L1H–L3H and their Cu(II) complexes were synthesized and characterized by the analytical and spectroscopic methods such as elemental analyses, infrared, mass, 1H and 13C NMR spectra. Molecular structures of the azo-azomethine ligands were determined by single crystal X-ray diffraction studies. X-ray data revealed that the compound L1H adopts azo-enamine tautomer form in the solid state while the compounds L2H and L3H favor azo-imine form. The harmonic oscillator model of aromaticity (HOMA) indexes for rings of synthesized azo-azomethine ligands were calculated in order to investigate of azo-enamine tautomer in the solid state. In L2H, the central phenol ring shows non-aromatic character with HOMA index value of 0.4872 suggesting the keto-amine tautomer in the solid state. All three azo-azomethine ligands involve in an intramolecular hydrogen bonding (NH⋯O for L1H and OH⋯N resulting in a S(6) graph set motif. The effect of solvents with different polarities on keto-enamine/phenol-imine tautomers was studied by UV-Vis spectroscopy. In the structure of the Cu(II) complexes, the central metal atom is coordinated to two phenolate oxygen atoms and two imine nitrogen atoms of two azo-azomethine molecules in a distorted square-planar geometry. All the synthesized azo-azomethine ligands and their Cu(II) complexes were screened for their cytotoxicity against HeLa (human uterus carcinoma) cell lines. Finally, antioxidant capacities of the compounds were determined by different antioxidant tests such as, free radical scavenging activity, metal chelating activity, and reduction power activity.

Bioactivity-Guided Isolation of Antiproliferative Sesquiterpene Lactones from Centaurea solstitialis L. ssp. solstitialis.

Centaurea solstitialis L. ssp. solstitialis (CSS) has been used as medicine for various diseases. In this work, root, stem and flower parts of the plant were separately extracted with methanol to execute bioassay-guided isolation. Antiproliferative activities of each extracts on C6 cells (Rat Brain tumor cells) and HeLa cells (human uterus carcinoma) were investigated in vitro. The methanol extract of stem exhibited the most antiproliferative activity therefore isolation of active compounds was carried out for stem of the plant. Methanol extract of stem was boiled at 97 °C for 2 hours in water and then hexane and ethyl acetate were extracted sequentially. Solstitialin A 1 and 15-dechloro-15-hydroxychlorojanerin 2 were isolated from ethyl acetate extract by column chromatography and identified by spectroscopic techniques. Solstitialin A 1 was isolated from CSS and 15-dechloro-15-hydroxychlorojanerin 2 was isolated from Saussurea lipschitz and Rhaponticum pulchrum previously. These two compounds exhibited very high antiproliferative activity on C6 and HeLa cells. IC50 and IC75 values of compound 1 were obtained as 10.78 and 53.65 against C6 cell and as 48.78 and 68.52 against HeLa, respectively. IC50 and IC75 values of compound 2 were determined as 432.43 and 109.79 against C6 cell.

Design, synthesis, characterization, and antiproliferative activity of novel pyrazole-3-carboxylic acid derivatives

In this study several novel 4-substituted-1-(3-nitrophenyl)-5-phenyl-1H-pyrazole-3-carboxylic acid derivatives were synthesized. The structures of these pyrazole-3-carboxylic acid derivatives were characterized by FT-IR, 1H NMR, 13C NMR, and elemental analysis methods. Synthesized molecules were screened to evaluate their antiproliferative activities against Vero (African green monkey kidney), C6 (rat brain tumor), and HeLa (human uterus carcinoma) cells as in vitro. The tests were carried out as dose-dependent assay starting from 100 to 500 μg/cm3. Ethyl 1-(3-nitrophenyl)-5-phenyl-3-(phenylcarbamoyl)-1H-pyrazole-4-carboxylate exhibited the highest performance against HeLa, Vero, and C6 cells among the synthesized derivatives. Also, it showed better antiproliferative activity compared with 5-fluorouracil (5-FU) against Vero cell. Two compounds showed moderate antiproliferative activities against C6 cell line when compared with 5-FU. Also moderate antiproliferative activities against Vero and HeLa cell lines were found.

Antiproliferative activities of isolated flavone glycosides and fatty acids from Stachys byzantina

From the aerial parts of Stachys byzanthina C. Koch., a new flavone glycoside, was isolated for the first time in addition to known two flavone glycosides. Structures were established by conventional methods of analysis and confirmed by 1H, 13C NMR and mass spectral analysis. Antiproliferative activities of isolated compounds, crude extract and fractions, fatty acids (extracts of hexane and hexane:ethyl acetate, 9:1) of aerial parts of S. byzantina were investigated against Vero (African green monkey kidney), HeLa (human uterus carcinoma) and C6 (rat brain tumor) cells in vitro and compared with 5-fluorouracil (5-FU). Antiproliferative effect of the extract, isolated flavonoids and fatty acids were tested at 100, 250, 500 and 1000μg/mL using BrdU cell proliferation ELISA.

Antiproliferative activity of pentadeca-(8E, 13Z) dien-11-yn-2-one and (E)-1,8-pentadecadiene from Echinacea pallida (Nutt.) Nutt. roots

Several species of Echinacea, a perennial plant which belongs to the Asteraceae family, possess medicinal properties and are currently used in phytotherapy. In the present study, antiproliferative activity of methanol extract and isolated structures of pentadeca-(8E, 13Z)-dien-11-yn-2-one 1 and (E)-1,8-pentadecadiene 2 from Echinacea pallida roots on C6 cells (Rat Brain tumor cells) and HeLa cells (human uterus carcinoma) was investigated in vitro. Antiproliferative effect of the extract, isolated compounds, and cisplatin were tested at 5, 10, 20, 30, 40, 50, 75, and 100 μg ml−1 using BrdU Cell Proliferation ELISA. The methanol extract and Compound 1 significantly inhibited proliferation of HeLa and C6 cancer cell lines.

Bioactive Volatile Content of the Stem and Root of Centaurea carduiformis DC. subsp. carduiformis var. carduiformis

Several species of Centaurea, biennial (or more rarely perennial) plants which belong to the Asteraceae family, possess medicinal properties and are currently used in phytotherapy. In the present study, antiproliferative activity of hexane extract from stems and roots of Centaurea carduiformis DC. subsp. carduiformis var. carduiformis (CCS and CCR) on Vero cells (African green monkey kidney), C6 cells (Rat Brain tumor cells), and HeLa cells (human uterus carcinoma) was investigated in vitro. Antiproliferative effect of the extract was tested at 500 μg/mL and 1000 μg/mL using BrDu Cell Proliferation ELISA. The hexane extract was significantly inhibited proliferation of Vero, HeLa, and C6 cancer cell lines with absorbance values. The extract of CCS and CCR showed the highest activity against the Vero, HeLa, and C6 cell lines at 500 μg/mL and 1000 μg/mL.

Synthesis, determination of the lipophilicity, anticancer and antimicrobial properties of some fused 1,2,4-triazole derivatives

3-Unsubstituted and 3-substituted-7-aryl-5 H-6,7-dihydroimidazo[2,1- c][1,2,4]triazoles ( 1– 14) were designed and obtained from biologically active 1-aryl-2-hydrazonoimidazolidines by cyclocondensation reaction with triethyl orthoformates ( 1– 4), phenoxyacetic acid derivatives ( 5– 13) and carbon disulfide ( 14), respectively. Their chemical structures were confirmed by IR, 1H NMR, 13C NMR, MS spectra and elemental analysis. In the high performance liquid chromatographic series of experiments, fourteen synthesized compounds ( 1– 14) were chromatographed on octadecyl silica adsorbent and their lipophilicity parameter (log k W) was determined using various aqueous systems: mixture of water and organic modifiers (methanol – MeOH, acetonitrile – MeCN or dioxane – DX). Compounds 7 and 12 were evaluated for their cytotoxic activity against three cancer cell lines: human Caucasian colon adenocarcinoma cell line – LS180 (ECACC 87021202), human uterus carcinoma cell line – SiHa (ECACC 85060701) and human breast carcinoma cell line – T47D (ECACC 85102201). Compound 12 was found to be the most effective in vitro against human colon adenocarcinoma cell line (LS180). Moreover, the distinctly marked lower cytotoxicity of compounds 7 and 12 against the normal cell line – human skin fibroblasts (HSF) and almost several-fold higher against the examined cancer cell lines was ascertained. The cytotoxic effect of imidazotriazole 7 was noticed on DNA structure of breast cancer cell line (T47D) by using the comet assay. Compound 7 in concentration of 29.3 μM was found to possess efficiency for DNA strand breakage. In particular, this led to cutting of the DNA strands and formation of small fragments of DNA – two higher and one lighter in comparison with control DNA. Moreover, significant viability decreases in the human leukaemic RPMI 8226 cells treated with different concentrations of imidazotriazoles 8– 12 were observed, suggesting their antiproliferative properties. Besides, three tested compounds ( 9, 13, 14) revealed significant antimicrobial activities with MIC values in the range of 30.9–44.0 μM. Compound 13 showed superior antibacterial activity to ampicillin and chloramphenicol in vitro, whereas 14 displayed superior antifungal activity to miconazole.

Synthesis, DNA Interaction, and Cytotoxic Activity of a Novel Proflavine−Dithiazolidinone Pharmacophore

Five novel proflavine-dithiazolidinone derivatives 4a-4e have been designed and synthesized by the reaction of dialkyl acridin-3,6-diyl dithioureas 3a-3e with methyl bromoacetate. The binding affinity of dithiazolidinone hydrochlorides 5a-5e with calf thymus DNA and plasmid (pUC19) DNA was investigated by a variety of spectroscopic techniques including UV-vis, fluorescence, and CD spectroscopy. The effects of 5a-5e on the thermal denaturation profiles of calf thymus DNA were also studied. From spectrophotometric and spectrofluorimetric titrations, the binding constants for the pUC19 DNA-drug complexes were determined (K = 6.2-2.2 x 104 M-1). In vitro cytotoxic activities of compounds 5a-5e toward murine leukemia cell line L1210 and human uterus carcinoma HeLa cells were also examined. 2',2' '-[(Acridin-3,6-diyl)diimino]-3',3' '-dipropyl-1,3-dithiazolidin-4-one hydrochloride (5b) showed the highest activity against these cells with IC50 values of 6.3 microM and 12.9 microM over the course of 72 h.

The G-Quadruplex-Interactive Molecule BRACO-19 Inhibits Tumor Growth, Consistent with Telomere Targeting and Interference with Telomerase Function

Interference with telomerase and telomere maintenance is emerging as an attractive target for anticancer therapies. Ligand-induced stabilization of G-quadruplex formation by the telomeric DNA single-stranded 3' overhang inhibits telomerase from catalyzing telomeric DNA synthesis and from capping telomeric ends. We report here the effects of a 3,6,9-trisubstituted acridine compound, BRACO-19, on telomerase function in vitro and in vivo. The biological activity of BRACO-19 was evaluated in the human uterus carcinoma cell line UXF1138L, which has very short telomeres (2.7 kb). In vitro, nuclear human telomerase reverse transcriptase (hTERT) expression was drastically decreased after 24 hours, induction of cellular senescence and complete cessation of growth was seen after 15 days, paralleled by telomere shortening of ca. 0.4 kb. In vivo, BRACO-19 was highly active as a single agent against early-stage (68 mm(3)) tumors in a s.c. growing xenograft model established from UXF1138L cells, if given chronically at 2 mg per kg per day i.p. BRACO-19 produced growth inhibition of 96% compared with controls accompanied by partial regressions (P < 0.018). Immunostaining of xenograft tissues showed that this response was paralleled by loss of nuclear hTERT protein expression and an increase in atypical mitoses indicative of telomere dysfunction. Cytoplasmic hTERT expression and its colocalization with ubiquitin was observed suggesting that hTERT is bound to ubiquitin and targeted for enhanced degradation upon BRACO-19 treatment. This is in accord with a model of induced displacement of telomerase from the telomere. The in vitro and in vivo data presented here is consistent with the G-quadruplex binding ligand BRACO-19 producing an anticancer effect by inhibiting the capping and catalytic functions of telomerase.

Antiproliferative constituents in plants 14. Coumarins and acridone alkaloids from Boenninghausenia japonica NAKAI.

The MeOH extracts of the ground part and the root of Boenninghausenia japonica NAKAI showed inhibitory activity against tumor cell growth. Fractionation of the extracts has resulted in isolation of 1,3-dihydroxy-4-(2'-hydroxy-3'-hydroxymethyl-3',4'-epoxy-butyl)-N-methylacridone, 1,3-dihydroxy-4-[(Z)-3'-hydroxy-3'-methyl-buten-1'-yl]-N-methylacridone, 3-(1',1'-dimethylallyl)-7-hydroxy-8-methoxy-2H-1-benzopyran-2-one, casegravol, cis-casegravol, and edgeworin in addition to 9 compounds reported from B. japonica and B. albiflora. The isolates from this plant and some related compounds were tested for antiproliferative activity against human gastric adenocarcinoma (MK-1), human uterus carcinoma (HeLa), and murine melanoma (B16F10) cells.