Determination of antioxidant, DNA protection, enzyme inhibition potential and molecular docking studies of a biomarker ursolic acid in Nepeta species.

Ursolic acid (UA), which has many biological properties such as anti-cancer, anti-inflammatory and antioxidant, and regulates some pharmacological processes, has been isolated from the flowers, leaves, berries and fruits of many plant species. In this work, UA was purified from the methanol-chloroform crude extract of Nepeta species (N. aristata, N. baytopii, N. italica, N. trachonitica, N. stenantha) using a silica gel column with chloroform or ethyl acetate solvents via bioactivity-guided isolation. The most active sub-fractions were determined under bioactivities using antioxidant and DNA protection activities and enzyme inhibitions. UA was purified from these fractions and its structure was elucidated by NMR spectroscopy techniques. The highest amount of UA was found in N. stenantha (8.53 mg UA/g), while the lowest amount of UA was found in N. trachonitica (1.92 mg UA/g). The bioactivities of UA were evaluated with antioxidant and DNA protection activities, enzyme inhibitions, kinetics and interactions. The inhibition values (IC50) of α-amylase, α-glucosidase, urease, CA, tyrosinase, lipase, AChE, and BChE were determined between 5.08 and 181.96 µM. In contrast, Ki values of enzyme inhibition kinetics were observed between 0.04 and 0.20 mM. In addition, Ki values of these enzymes for enzyme-UA interactions were calculated as 0.38, 0.86, 0.45, 1.01, 0.23, 0.41, 0.01 and 2.24 µM, respectively. It is supported that UA can be widely used as a good antioxidant against oxidative damage, an effective DNA protector against genetic diseases, and a suitable inhibitor for metabolizing enzymes.Communicated by Ramaswamy H. Sarma.

In vitro anticancer, antioxidant and enzyme inhibitory potentials of endemic Cephalaria elazigensis var. purpurea with in silico studies.

In this study, the therapeutic potential and phytochemical composition of ethanolic extract of Cephalaria elazigensis var. purpurea (CE), an endemic species, were investigated. For this purpose, the antiproliferative effect of CE on the MCF-7 human breast cancer cell line was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and its effectiveness on colony formation and cell migration was analyzed with clonogenic assay and wound healing assay, respectively. In addition, the cell death detection ELISA (CDDE) assay was conducted to determine the pro-apoptotic capacity of CE. The IC50 value of the CE was determined as 324.2 ± 14.7 µg/mL. Furthermore, upon 1000 µg/mL CE treatment, there was 4.96-fold increase in the population of cells undergoing apoptosis compared to the untreated control cells. The antioxidant activity tests were performed by DPPH free radical, ABTS cation radical, ferric-ion reducing power (FRAP) and ferrous-ion chelating power (FCAP) assays. Antioxidant activity values for the DPPH, ABTS and FRAP assays were found to be 125.6 ± 6.3, 34.09 ± 0.1 and 123.4 ± 4.2 µmol TE/mg DE, respectively. We further determined the effect of CE ethanolic extract against acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) enzymes. CE plays an effective inhibitory role in AChE and BuChE (AChE: IC50: 10.54 µg/mL, BuChE: IC50: 6.84 µg/mL) respectively. Further, molecular docking stuy was conducted to understand the nature of the all compound against AChE an BChE. It is revealed that α-Linolenic acid shows lowest binding energy (-7.90 kcal/mol) towards AChE, on the other side, Linoleic acid shows good binding affinity (-7.40 kcal/mol) for BChE.Communicated by Ramaswamy H. Sarma.

Three new iridoids from the aerial parts of Nepeta teucriifolia Willd. and antiproliferative activities of extracts.

The aerial parts of Nepeta teucriifolia Willd. were extracted with the solvents of different polarities. The antiproliferative activities of the extracts were evaluated against rat brain tumor (C6) and human cervix carcinoma (HeLa) cell lines. The phytochemical screening of the extracts was performed with TOF-LC/MS. The CH2Cl2 and EtOAc extracts showed considerable antiproliferative activities against HeLa cells at higher concentration (250 µg mL-1). The CH2Cl2 extract was found more active than the others on both cells. The phytochemical studies of the active extract led to the isolation of three new iridoids, teucriifolian A-C (1-3). The structure elucidations of the new compounds were performed using HPLC-TOF/MS, 1D and 2D NMR techniques. The compounds 1-3 were evaluated in terms of their antiproliferative activities against HeLa and C6 cells, respectively. The results indicated that only 2 had moderate antiproliferative activity against HeLa cells at 250 µg mL-1.

Fatty acid composition, enzyme inhibitory effect, antioxidant and anticancer activity of extract from Saponaria prostrata WILLD. subsp. anatolica HEDGE.

This study attempts to evaluate the antioxidant, enzyme inhibitory, and anticancer properties as well as fatty acid compositions of endemic Saponaria prostrata WILLD. subsp. anatolica HEDGE. The gas chromatography-mass spectrometry (GC-MS) was used to determine the fatty acid content of methanol: dichloromethane extract from S. prostrata subsp. anatolica (SPA). Enzymatic activity was measured against acetylcholinesterase, butyrylcholinesterase and α-glucosidase. DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging activity and Ferric reducing antioxidant power assay (FRAP) were conducted to antioxidant properties. The anticancer effect of SPA on human MCF-7 breast cancer and human HCT116 colorectal cancer cell line was evaluated by WST-1 cell viability assay, colony formation assay and wound healing assay. In addition, human VEGF Elisa method was used to determine the anti-angiogenic effect, and the quantitative real-time PCR (qRT-PCR) method on p53, Bax and Bcl-2 mRNA levels were used to evaluate apoptosis. While high amounts of palmitic acid (40.8%), linoleic acid (17.75%) and α-linolenic acid (16.84%) were detected in the SPA, the total amount of unsaturated fatty acid (51.34%) was higher than the total amount of saturated fatty acid (48.66%). SPA displayed the most promising acetylcholinesterase (AChE), butyrylcholinesterase (BuChE) and α-glycosidase (AG) inhibitory activities (AChE: IC50: 18.03 µg/mL, BuChE: IC50: 44.24 µg/mL and AG: IC50: 210.85 µg/mL). The half maximum inhibitory concentration (IC50) of SPA in MCF-7 and HCT116 cells was determined as 259.79 µg/mL and 97.24 µg/mL, respectively. In addition, it was determined that SPA suppresses colony formation and wound closure, and suppresses angiogenesis as well as triggering apoptosis at a significant level. It is true that endemic S. prostrata subsp. anatolica is a potential source of functional food ingredients, but more analytical and in vivo experiments are needed to explore further secondary metabolite diversity and pharmacological properties.

The cytotoxicity and antioxidant activity analysis of the isolated constituents and extracts from endemic Centaurea derderiifolia.

It is aimed to investigate the cytotoxic effects of the various extracts from the leaf and seed of Centaurea derderiifolia on the growth human cervical adenocarcinoma (HeLa) cells by xCELLigence method and to isolate the cytotoxic constituents. The results showed that subfractions 2 and 3 exhibited remarkable inhibitory effect against HeLa (IC50 < 10 µg/mL). The chloroform extract of leaf displayed the highest cytotoxic effect on HeLa cells (IC50 < 50 µg/mL) and was therefore subjected to a bioassay-guided multistep separation procedure. The pure compounds were elucidated by spectroscopic and mass-spectrometric analyses, including 1 D-, 2 D-NMR. In addition to cytotoxic effects of the isolated constituents, their antioxidant activities were also studied. On the other hand, subfraction 4 exhibited the highest DPPH radical scavenging activity (IC50 = 0.76 ± 0.03 mg/mL). ß-sitosterol-3-O-ß-d-glucopyranoside was isolated for the first time from this plant and three compounds from the bioactive subfractions were identified.

Phytochemical Screening, Antiproliferative and Antioxidant Properties of Various Extracts from Endemic Origanum acutidens.

AIM AND OBJECTIVE: Origanum acutidens (Hand.-Mazz.) Ietsw. is an endemic and perennial plant grown mainly in East Anatolia. Recently, natural plant products have attracted interest due to their safety and therapeutic effects. Therefore, the aim of this study was to investigate phytochemical contents and biological effects of Origanum acutidens. MATERIALS AND METHODS: The aerial parts of O. acutidens were extracted with water, ethyl acetate, nbutanol, and methanol/chloroform solvents. Phenolic compounds and other constituents of the extracts were analyzed by HPLC/TOF-MS. The Ethyl Acetate extract (EA) was fractionated by flash chromatography. The extracts and fractions were investigated for their antiproliferative activities on human cervical adenocarcinoma (HeLa) cell line by using BrdU ELISA assay. Antioxidant activities of the extracts and fractions were evaluated by complementary test systems, namely determination of total phenolic contents, metal chelating ability and DPPH radical scavenging assay. RESULTS: Among the extracts, Ethyl Acetate extract (EA) exhibited the highest antiproliferative activity (IC50 = 15.71 ± 0.04 µg/mL) on HeLa cells. It was therefore fractionated by flash chromatography to obtain 10 fractions which were investigated for their phenolic compounds and bioactivities. Rosmarinic acid was determined as the major component of EA and its fractions. EA exhibited higher antiproliferative activity against HeLa cell line than its fractions and 5-fluorouracil (5-FU) at the concentration of 100 µg/mL. EA and its fractions (F10, F6, F4, F7, F3, and F2) displayed higher radical scavenging activity compared to Butylated Hydroxytoluene (BHT). These effects may be attributed to the presence of rosmarinic acid in EA and its active fractions. CONCLUSION: This study demonstrated that O. acutidens is an essential natural source of polyphenols and a potent natural antioxidant and antiproliferative agent for food and pharmaceutical industries.

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-ß-xylopyranosyl-(1-2)-ß-rhamnopyranoside (1) along with five known compounds, quercetin-3-O-ß-xyloside (2), kaempferol-3-O-ß-glucoside (3), quercetin-3-O-ß-glucoside (4), kaempferol-3-O-ß-rhamnopyranoside (5), and kaempferol-3-O-ß-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.

Phytochemical analysis, antiproliferative and antioxidant activities of Chrozophora tinctoria: a natural dye plant.

CONTEXT: Chrozophora tinctoria (L.) A. Juss. (Euphorbiaceae) is known as 'dyer's-croton' and used to obtain dye substances. Recently, natural antioxidants and colorants have been of interest because of their safety and therapeutic effects. OBJECTIVE: This study investigates the antiproliferative and antioxidant activities of the various extracts and fractions from C. tinctoria and analyzes their phytochemical contents. MATERIALS AND METHODS: The aerial parts of C. tinctoria were extracted with water, ethyl acetate, n-butanol, and methanol/chloroform. Phenolic compounds and other constituents of the extracts were analyzed by HPLC/TOF-MS. The ethyl acetate extract (EA) was fractionated by flash chromatography. The extracts, fractions, and major phenolic compounds were investigated for their antiproliferative activities on human cervical adenocarcinoma (HeLa) cell line at the concentrations of 5-100 µg/mL by using BrdU ELISA assay during 24 h of incubation. DPPH radical scavenging activities (5-150 µg/mL) and total phenolic contents of the samples were also evaluated. RESULTS: 4-Hydroxybenzoic acid (268.20 mg/kg), apigenin-7-glucoside (133.34 mg/kg), and gallic acid (68.92 mg/kg) were the major components of EA. CT/E-F6 (IC50 = 64.59 ± 0.01 µg/mL) exhibited the highest antiproliferative activity. CT/E-F2 (IC50= 14.0 ± 0.0 µg/mL) and some fractions displayed higher radical scavenging activity compared to synthetic antioxidant BHT (IC50 = 23.1 ± 0.0 µg/mL). Among the main phenolics, gallic acid exhibited the highest antiproliferative and radical scavenging abilities (IC50 < 5 µg/mL). CONCLUSION: In this study, we have determined the biologically active fractions and their high effects may be attributed to the presence of gallic acid.

Phytochemical screening, anticancer and antioxidant activities of Origanum vulgare L. ssp. viride (Boiss.) Hayek, a plant of traditional usage.

BACKGROUND: A detailed phytochemical analysis of Origanum vulgare L. ssp. viride (Boiss.) Hayek was carried out and the antioxidant activities of five different crude extracts were determined. The antiproliferative activities of the extracts were determined using the xCELLigence system (Real Time Cell Analyzer). RESULTS: Differences between the essential oil and volatile organic compound profiles of the plant were shown. The main component of the essential oil was caryophyllene oxide, while the main volatile organic compounds were sabinene and eucalyptol as determined by HS-GC/MS. Phenolic contents of the extracts were determined qualitatively and quantitatively by HPLC/TOF-MS. Ten phenolic compounds were found in the extracts from O. vulgare and Origanum acutidens: rosmarinic acid (in highest abundance), chicoric acid, caffeic acid, p-coumaric acid, gallic acid, quercetin, apigenin-7-glucoside, kaempferol, naringenin and 4-hydroxybenzaldehyde. CONCLUSION: This study provides first results on the antiproliferative and antioxidant properties and detailed phytochemical screening of O. vulgare ssp. viride (Boiss.) Hayek.