Solvent-partitioned fractions from <Emphasis Type="Italic">Ishige okamurae</Emphasis> extract inhibit MMP-2 and MMP-9 activities in human fibrosarcoma cells in vitro

Matrix metalloproteinases (MMPs) are endopeptidases that take significant roles in extracellular matrix degradation and therefore linked with several complications such as metastasis of cancer progression, oxidative stress, and hepatic fibrosis. Ishige okamurae, a brown alga, has been reported to possess various bioactivities including but not limited to antiviral, antimicrobial, and anti-inflammatory. In this study, the potential of I. okamurae against cancer cell invasion was evaluated through MMP inhibitory effect in HT1080 fibrosarcoma cells in vitro. I. okamurae crude extract was fractionated with organic solvents, namely water (H₂O), n-butanol (n-BuOH), 85% aqueous methanol (85% aq. MeOH), and n-hexane (n-Hex). The non-toxicity of fractions was confirmed by MTT assay. All fractions inhibited the enzymatic activities of MMP-2 and MMP-9 according to gelatin zymography assay. Cell migration was also inhibited by 85% aq. MeOH fraction, significantly. In addition, both gene and protein expressions of MMP-2 and MMP-9, and tissue inhibitor of MMPs (TIMPs) were evaluated by RT-PCR and Western blotting, respectively. Fractions suppressed the mRNA and protein levels of MMP-2 and MMP-9 while elevating the TIMP-1 and TIMP-2, H₂O fraction being the least effective while 85% aq. MeOH fraction the most. Collectively, 85% aq. MeOH fraction from brown algae could be a potential inhibitor of MMPs, suggestively considering presence of poly-unsaturated fatty acids in high amounts.     

Dual effects of isoflavonoids from Pueraria lobata roots on estrogenic activity and anti-proliferation of MCF-7 human breast carcinoma cells

Pueraria lobata root (PLR), well known as Kudzu root, has recently become commercially available in Western dietary supplements for menopausal symptoms. The scientific basis for its action has been attributed to the action of phytoestrogens. This study aimed to investigate the estrogen-like activity of isoflavonoids isolated from P. lobata root and their safety with respect to their effect on breast cancer cell proliferation. In an E-screen assay, crude MeOH extract of PLR significantly increased the proliferation of MCF-7 cells in a concentration-dependent manner. Among the four fractions obtained by solvent fractionation of MeOH extract, the n-BuOH fraction had significant estrogen-like activities at all concentrations tested. Phytochemical analysis of the n-BuOH fraction led to the isolation of 10 isoflavones (1–10), among which genistein (10) had significant estrogen-like activities at all concentrations tested. These activities were significantly enhanced by treatment with genistein and 17β-estradiol compared with 17β-estradiol alone, and this effect was mediated by decreased expression of estrogen receptor (ER)α and phospho-ERα in MCF-7 cells. In a cell cytotoxicity assay, genistein (10) exhibited significant cytotoxicity in both ER-positive MCF-7 and ER-negative MDA-MB-231 breast cancer cells. This cytotoxicity was characterized by the induction of apoptotic cells stained with annexin V conjugated with Alexa Fluor 488 and involved activation of mitochondria-independent and -dependent apoptosis pathways in MCF-7 cells. Our results demonstrated that genistein (10) has estrogen-like effects dependent on ER pathway activation and anti-proliferative effects mediated by the apoptosis pathway rather than the ER pathway in MCF-7 breast cancer cells.     

Antioxidant activity of the chemical constituents from the flower buds of <Emphasis Type="Italic">Magnolia denudata</Emphasis>

The scavenging activity of the flower buds of Magnolia denudata Desrousseaux on reactive oxygen species (ROS) was evaluated using 2′,7′-dichlorofluorescin diacetate (DCFH-DA) in HT 1080 cells. Methanol (MeOH) and dichloromethane (CH₂Cl₂) extracts inhibited dose-dependently generation of ROS in the cellular system. MeOH and CH₂Cl₂ extracts were combined and fractionated with n-hexane, 85% aqueous MeOH, and n-butanol (n-BuOH). Both n-hexane-soluble and 85% aqueous-soluble fractions showing strong radical-scavenging activity in the cellular system were further separated by diverse chromatographic methods to give five known lignans (1–5). All these compounds exhibited significant radical-scavenging effect on intracellular ROS in a dose-dependent manner. Their scavenging activity on various reactive oxygen species (ROS) was also evaluated using electron spin resonance (ESR) spin-trap techniques.     

Isolation of diverse bioactive compounds from Euphorbia balsamifera: Cytotoxicity and antibacterial activity studies

Antibacterial and cytotoxic activities of Euphorbia balsamifera, fractions and pure compounds were evaluated. The cytotoxic assays for HCT116, HePG2 and MCF7 showed a significant IC₅₀: 54.7 and 76.2 µg/mL of non-polar fraction “n-hexane” against HCT116 and HePG2, respectively. Antibacterial results revealed that plant fractions exhibited significant potential against the tested pathogens than the total extract where n-butanol and ethyl acetate fractions showed significant antibacterial activity (P < 0.05) against tested bacterial strains. Isolation and structure determination of compounds from n-hexane and n-butanol fractions were performed. From n-hexane fraction, 29-nor-cycloartanol (1), lanost-8-en-3-ol (2a), cycloartanol (2b) and kampferol-3,4'-dimethyl ether (3) were isolated and structurally identified, along with 24 compounds were tentatively identified by GC–MS. From the polar n-butanol fraction, 4-O-β-D-glucopyranosyl-2-hydroxy-6-methoxyacetophenone (4), 4-O-α-L-rhamnosyl-(1 → 6)-β-D-glucopyranosyl-2-hydroxy-6methoxy-acetophenone (5), quercetin-3-O-glucopyranoside (6) and isoorientin (7) were assigned. Structures of the obtained compounds were determined by nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry. Except compounds 1 and 5, all reported compounds announced antibacterial efficiency. Compound 2 showed selectively the highest activity against Enterococcus faecalis (22 ± 0.13 mm), meanwhile 4-O-β-D-glucopyranosyl-2-hydroxy-6-methoxyacetophenone (4) showed broadly the highest antibacterial activity with MIC of 1.15–1.88 mg/mL against the test Gram-positive and Gram-negative bacteria. Cytotoxic assays indicated that kampferol-3,4'-dimethyl ether (3) exhibited the highest activity with matching IC₅₀ values to doxorubicin; 111.46, 42.67 and 44.90 µM against HCT116, HePG2 and MCF7, respectively, however, it is toxic on retina normal cell line RPE1.     

A novel tyrosine kinase activity in the cotton leafworm,Spodoptera littoralis

A rapid, highly-specific ELISA for tyrosine kinases readily detects such activity in crude extracts prepared from rat mammary epithelial and fibroblastic cells that have been stimulated with epidermal growth factor or basic fibroblast growth factor. Tyrosine kinase activity is also found in extracts of pupae of the cotton leafworm, Spodoptera littoralis. Both the mammalian and the insect tyrosine kinases are ATP-dependent. Both cytosol and membrane-associated (Triton-X-100-soluble) fractions of Spodoptera littoralis pupae contain tyrosine kinase activity. The growth factor-stimulated tyrosine kinase activities in extracts of growth factor-stimulated rat mammary cells are inhibited by genistein and an analogue of erbstatin: methyl 2,5-dihydroxycinnamate. However, the tyrosine kinase activities present in pupae of Spodoptera littoralis are not sensitive to these inhibitors, suggesting that the major tyrosine kinases of Spodoptera littoralis pupae may be distinct from the growth factor-stimulated mammalian tyrosine kinases.     

Identification of potential and selective collagenase,gelatinase inhibitors from Crataegus pinnatifida

Four oligomeric procyanidins were isolated from the MeOH extracts of the leaves of Crataegus pinnatifida (Rosaceae). Investigation of collagenase and gelatinase inhibitory natural components afforded four oligomeric procyanidins. Of these, 2 and 3 exhibited collagenase inhibitory activity (IC₅₀) at a concentration of less than 1 μM, and 3 showed gelatinases A and B inhibitory activity (IC₅₀) at 0.4 and 2.3 μM, respectively.     

A prenylated xanthone and antiproliferative compounds from leaves of Pentadesma butyracea

A new prenylated xanthone, butyraxanthone F (1) together with 20 known compounds were isolated from the leaves of Pentadesma butyracea. Their identification was achieved by spectroscopic means and comparison of the data with literature or by direct comparison on TLC with authentic samples. To the best of our knowledge, this is the first report of 2-hydroxy-2,3-dihydrosqualene (17) from a natural source. Compounds 2–6 and 11–16 were found for the first time in this genus. The antiproliferative activity of the crude extract, fractions and some isolated compounds was evaluated against three human cancer cell lines, BGC-823, Hela and A549. The crude extract, fractions B and C showed weak activities. Globuxanthone (2) and 30-epi-cambogin (7) were found to be the most active compounds on the three cancer cells. The effect of 2 on Hela cells and that of 7 on BGC-823 cells were noteworthy. The antiproliferative potential of allanxanthone A (3), ergosta-4,6,8(14),22-tetraen-3-one (14) and 2-hydroxy-2,3-dihydrosqualene (17) against BGC-823 and Hela cells could be classified as moderate-to-weak.     

Macrocyclic diterpenes resensitizing multidrug resistant phenotypes

Herein, collateral sensitivity effect was exploited as a strategy to select effective compounds to overcome multidrug resistance in cancer. Thus, eleven macrocyclic diterpenes, namely jolkinol D (1), isolated from Euphorbia piscatoria, and its derivatives (2–11) were evaluated for their activity on three different Human cancer entities: gastric (EPG85-257), pancreatic (EPP85-181) and colon (HT-29) each with a variant selected for resistance to mitoxantrone (EPG85-257RN; EPP85-181RN; HT-29RN) and one to daunorubicin (EPG85-257RD; EPP85-181RD; HT-29RD). Jolkinol D (1) and most of its derivatives (2–11) exhibited significant collateral sensitivity effect towards the cell lines EPG85-257RN (associated with P-glycoprotein overexpression) and HT-29RD (altered topoisomerase II expression). The benzoyl derivative, jolkinoate L (8) demonstrated ability to target different cellular contexts with concomitant high antiproliferative activity. These compounds were previously assessed as P-glycoprotein modulators, at non-cytotoxic doses, on MDR1-mouse lymphoma cells. A regression analysis between the antiproliferative activity presented herein and the previously assessed P-glycoprotein modulatory effect showed a strong relation between the compounds that presented both high P-glycoprotein modulation and cytotoxicity.     

UHPLC-MS/MS-GNPS based phytochemical investigation of Equisetum arvense L. And evaluation of cytotoxicity against human melanoma and ovarian cancer cells

Equisetum arvense L. is widely used as a traditional medicine for the management of inflammation and cancer. In the present study, phyto-chemical analysis of E. arvense was carried out and its cytotoxic potential against human melanoma (MDA-MB-435) and ovarian cancer cells (OVCAR3) was evaluated. Phyto-chemical profile of E. arvense methanolic extract and its fractions was established employing UHPLC-MS/MS and Global Natural Product Social molecular networking. Cytotoxic activity was evaluated using absorbance assay (CellTiter-Blue® Cell Viability Assay). Overall, 22 compounds were identified in the crude extract and polarity-based fractions of E. arvense. Flavonoids, flavonoid-O-glycosides and phenolic acids were found to be the major classes of phyto-chemicals. In addition, the crude extract of E. arvense and its fractions were found active against the tested cell lines. The highest anti-cancer activity against OVCAR3 cells was exhibited by the n-hexane fraction. These results indicated that E. arvense is rich in flavonoids and might be used for the development of anti-cancer drugs against melanoma and ovarian cancers.     

Seasonal variation in phenolic composition, antibacterial and antioxidant activities of Ulva rigida (Chlorophyta) and assessment of antiacetylcholinesterase potential

The phenolic composition and antibacterial and antioxidant activities of the green alga Ulva rigida collected monthly for 12 months were investigated. Significant differences in antibacterial activity were observed during the year with the highest inhibitory effect in samples collected during spring and summer. The highest free radical scavenging activity and phenolic content were detected in U. rigida extracts collected in late winter (February) and early spring (March). The investigation of the biological properties of U. rigida fractions collected in spring (April) revealed strong antimicrobial and antioxidant activities. Ethyl acetate and n-hexane fractions exhibited substantial acetylcholinesterase inhibitory capacity with EC₅₀ of 6.08 and 7.6 μg mL^?1, respectively. The total lipid, protein, ash, and individual fatty acid contents of U. rigida were investigated. The four most abundant fatty acids were palmitic, oleic, linolenic, and eicosenoic acids.     

Bioactivity-based analysis and chemical characterization of anti-inflammatory compounds from Curcuma zedoaria rhizomes using LPS-stimulated RAW264.7 cells

Inflammation is not only a self-defense response of the innate immune system, but also the pathogenesis mechanism of multiple diseases such as arthritis, neurodegeneration, and cancer. Curcuma zedoaria Roscoe (Zingiberaceae), an indigenous plant of India, has been used traditionally in Ayurveda and folk medicine. As part of our ongoing efforts to screen traditional medicinal plants exhibiting pharmacological potential and to characterize the compounds involved, we examined the anti-inflammatory effects of the MeOH extract of C. zedoaria rhizomes using lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophage cells and found that MeOH extract inhibited the synthesis of nitric oxide (NO) in a dose-dependent manner (IC₅₀: 23.44 ± 0.77 μg/mL). In our efforts to characterize the compounds responsible for these anti-inflammatory effects, bioactivity-guided fractionation of the MeOH extract and chemical investigation of its active hexane-soluble fraction led to the successful isolation of five sesquiterpenes (1–5), the structures of which were elucidated by NMR spectroscopic analysis and LC/MS analysis. Among them, curcuzedoalide (5) exhibited potent inhibitory effects on NO synthesis (IC₅₀: 12.21 ± 1.67 μM) and also suppressed pre-inflammatory protein expression of iNOS and COX-2. Curcuzedoalide (5) was thus determined to be a contributor to the anti-inflammatory effect of C. zedoaria rhizomes and could be a potential candidate for therapeutic applications.     

Design,synthesis, and biological evaluation of novel aminopyrimidinylisoindolines as AXL kinase inhibitors

A novel series of aminopyrimidinylisoindoline derivatives 1a-w having an aminopyrimidine scaffold as a hinge region binding motif were designed and synthesized. Among them, six compounds showed potent inhibitory activities against AXL kinase with IC₅₀ values of submicromolar range. Especially, compound 1u possessing (4-acetylpiperazin-1-yl)phenyl moiety exhibited extremely excellent efficacy (IC₅₀?=?<0.00050?μM). Their in vitro antiproliferative activities were tested over five cancer cell lines. Most compounds showed good antiproliferative activities against HeLa cell line. The kinase panel profiling of 50 different kinases and the selected inhibitory activities for the representative compound 1u were carried out. The compound 1u exhibited excellent inhibitory activities (IC₅₀?=?<0.00050, 0.025, and 0.050?μM for AXL, MER, and TYRO3, respectively) against TAM family, together with potent antiproliferative activity against MV4-11 cell line (GI₅₀?=?0.10?μM) related to acute myeloid leukemia (AML).     

Ceramide,cerebroside and triterpenoid saponin from the bark of aerial roots of Ficus elastica (Moraceae)

Three compounds, ficusamide (1), ficusoside (2) and elasticoside (3), were isolated from the bark of aerial roots of Ficus elastica (Moraceae), together with nine known compounds, including four triterpenes, three steroids and two aliphatic linear alcohols. The chemical structures of the three compounds were established by extensive 1D and 2D NMR spectroscopy, mass spectrometry and by comparison with published data. The growth inhibitory effect of the crude extract and isolated compounds was evaluated against several microorganisms and fungi. The cytotoxicity against human cancer cell lines was also assessed. Ficusamide (1) displayed a moderate in vitro growth inhibitory activity against the human A549 lung cancer cell line and a strong activity against Staphylococcus saprophyticus, while elasticoside (3) showed a potent activity on Enterococcus faecalis.     

Antioxidant activity of the halophyte Limonium tetragonum and its major active components

In this study, the antioxidant potentials of crude extracts and solvent-partitioned fractions of Limonium tetragonum were assessed by measuring their ability to scavenge intracellular reactive oxygen species (ROS) generated in HT-1080 cells. Following activity-oriented separation, four flavonol glycosides were isolated as active principles and their chemical structures were determined by 2 D NMR and by comparison with reported spectral data. The isolated compounds (1?C4) were evaluated for their antioxidant capacity using three different activity tests; degree of occurrence of intracellular ROS, lipid peroxidation in HT-1080 cells and the extent of oxidative damage of genomic DNA purified from HT-1080 cells. All compounds exhibited significantly inhibited the generation of intracellular ROS and lipid peroxidation in HT-1080 cells, and significantly inhibited DNA oxidation. In addition, direct free radical scavenging effects of these compounds were investigated using the electron spin resonance (ESR) spin-trap technique.     

Separation and purification of bioactive botrallin and TMC-264 by a combination of HSCCC and semi-preparative HPLC from endophytic fungus Hyalodendriella sp. Ponipodef12

Two dibenzo-α-pyrones, botrallin (1) and TMC-264 (2) were preparatively separated from crude ethyl acetate extract of the endophytic fungus Hyalodendriella sp. Ponipodef12, which was isolated from the hybrid ‘Neva’ of Populus deltoides Marsh × P. nigra L. using a combination of high-speed counter-current chromatography (HSCCC) and semi-preparative HPLC. Botrallin (1) with 74.73 % of purity and TMC-264 (2) with 82.29 % of purity were obtained through HSCCC by employing a solvent system containing n-hexane–ethyl acetate–methanol–water at a volume ratio of 1.2:1.0:0.9:1.0. It was the first time for TMC-264 (2) to be isolated from this fungus. TMC-264 (2) showed strong antimicrobial and antinematodal activity, and botrallin (1) exhibited moderate inhibitory activity on acetylcholinesterase.     

Ent-kaurane diterpenes isolated from n-hexane extract of Baccharis sphenophylla by bioactivity-guided fractionation target the acidocalcisomes in Trypanosoma cruzi

BackgroundIn the present work the bioactivity-guided fractionation of n-hexane extract from aerial parts of Baccharis sphenophylla (Asteraceae) against trypomastigote forms of Trypanosoma cruzi was performed.PurposeTo evaluate the antitrypanosomal potential of diterpenes ent‑kaurenoic (1), grandifloric (2). and 15β-tiglinoyloxy‑ent-kaurenoic (3) acids, isolated from n-hexane extract from aerial parts of B. sphenophylla, and elucidate their mechanism of action against T. cruzi.Methods/Study designn-Hexane and MeOH extracts from aerial parts of B. sphenophylla were prepared and caused, respectively, 100% and 50% of death of trypomastigote forms of T. cruzi. Based on these results, the n-hexane extract was subjected to bioactivity-guided fractionation procedures to afford three related ent‑kaurane diterpenoids (1–3). Based on spectrofluorometric assays and flow cytometry analysis, the mechanism of action of compounds 1 and 3 was investigated.ResultsCompounds 1 and 3, isolated from n-hexane extract from aerial parts of B. sphenophylla, showed potent activity against parasites with EC₅₀ values of 10.6 μM (SI > 18.8) and 2.4 μM (SI = 34.8), respectively. On the other hand, compound 2 was inactive against trypomastigotes. In mechanism of action studies using the fluorescent probe SYTOX Green, the plasma membrane permeability was unaltered after treatment with compounds 1 and 3, but compound 1 induced a depolarization of the plasma membrane electric potential (ΔΨp). No substantial alterations were observed in the mitochondria after treatment with compound 3, but a transient hyperpolarization of the mitochondrial membrane potential (ΔΨm) by compound 1. Despite the increased ATP levels induced by compounds 1 and 3, no alterations of ROS and Ca²⁺ levels were registered. However, both compounds promoted a time-dependent alkalinization of the acidocalcisomes, probably contributing to an osmotic imbalance of the cell. In silico physicochemical studies of compounds 1–3 suggested that lipophilicity and molecular complexity may play an important role in the antitrypanosomal activity. Moreover, no pan-assay interference compounds (PAINS) alerts were detected for compounds 1–3.ConclusionObtained data indicated that the isolated ent‑kaurane diterpenes from n-hexane extract from aerial parts of B. sphenophylla, especially compound 3, could be considered interesting prototypes for further modifications aiming the discovery of new hits against T. cruzi.     

Design,synthesis and biological evaluation of novel chromeno[4,3-c]pyrazol-4(2H)-one derivates containing sulfonamido as potential PI3Kα inhibitors

A series of novel chromeno[4,3-c]pyrazol-4(2H)-one derivates contained sulfonamido were designed and synthesized, and their anticancer effects in vitro was evaluated to develop some new PI3Kα inhibitors. Most of desired compounds exhibited the better antiproliferative activities against four cancer cell lines than that of LY294002. Out of them, compound 4o displayed the potent antiproliferative activity and high selectivity against the PI3Kα protein and it can induce apoptosis of HCT116 in a dose-dependent manner. Western blot assay indicated that compound 4o obviously down-regulated expression of p-Akt (S473). Molecular docking was performed to clarify the possible binding mode between compound 4o and PI3Kα. All these results indicated that compound 4o could be a potential inhibitor of PI3Kα.     

Antioxidant properties ofErigeron annuus extract and its three phenolic constituents

The antioxidant activity of the extract ofErigeron annuus was assessed by means of two differentin vitro tests: bleaching of the stable 1,1-diphenyl-2-picrylhydrazyl radical (DPPH test) and the scavenging of authentic peroxynitrite in company with peroxynitrite generation from 3-morpholinosydnonimine (SIN-1). In both tests, the 85% aq. MeOH andn-BuOH soluble fractions of the crude extract showed a significant scavenging effect on peroxynitrite and DPPH radical in comparison to L-ascorbic acid. And bioassay-guided fractionation of then-BuOH soluble fraction led to the isolation of three compounds: Apigenin (1), quercetin-3-O-glucoside (2), and caffeic acid (3). The structures of the isolated compounds were elucidated on the basis of their spectroscopic data and their antioxidant activities were measured by determining their capacity to scavenge peroxynitrite and the DPPH radical.     

Design,synthesis, and biological evaluation of 4-phenoxyquinoline derivatives as potent c-Met kinase inhibitor

A series of novel 4-phenoxyquinoline derivatives containing 3-oxo-3,4-dihydro-quinoxaline moiety were synthesized and evaluated for their antiproliferative activity against five human cancer cell lines (A549, H460, HT-29, MKN-45 and U87MG) in vitro. Most of the tested compounds exhibited more potent inhibitory activities than the positive control foretinib. Compound 1b, 1s and 1t were further examined for their inhibitory activity against c-Met kinase. The most promising compound 1s (with c-Met IC₅₀ value of 1.42 nM) showed remarkable cytotoxicity against A549, H460, HT-29, MKN45 and U87MG cell lines with IC₅₀ values of 0.39 μM, 0.18 μM, 0.38 μM, 0.81 μM, respectively. Their preliminary structure-activity relationships (SARs) study indicated that the replacement of the aromatic ring with the cyclohexane improved their antiproliferative activity.