ABSTRACT
The peptides produced enzymatically from various plants have shown various biological activities including cytotoxicity. Different types of cytotoxic peptides have been reported from the seeds and leaves of Violaceae, Rubiaceae and Annonaceae families. In this study, we report purification and characterization of peptide(s) showing cytotoxic activity against A549 and HeLa cancer cell lines from the seeds of Polyalthia longifolia (Annonaceae). Seed proteins of P. longifolia were extracted and hydrolyzed using trypsin. The enzyme hydrolysate was applied on to a Sephadex G10 column and eluted using Tris-HCl buffer (pH 7.5). Two fractions F1 and F2 were obtained, of which F2 showed significant cytotoxic activity against lung (A549) cancer cells at 10 µg/mL and cervical (HeLa) cancer cell lines at 30 µg/mL, as revealed by the MTT assay. DNA fragmentation was observed in the tested cancer cell lines treated with F2 peptide at a concentration of 10 µg/mL and 30 µg/mL, respectively. Further, increased number of apoptotic cells was observed in sub-G0 phase of cell cycle of A549 and HeLa cell lines, when treated with 10 µg/mL and 30 µg/mL of F2, as revealed by the flow cytometric analyses. FTIR spectrum of F2 peptide detected the presence of stretching vibrations of carboxylic acid OH residue with peak at 3420 cm-1 and carbonyl (C=O) groups at 1636 cm-1, respectively. RP-HPLC analysis of F2 peptide showed a single peak at a retention time of 12.8 min detected at 280 nm, depicting the purity of F2 to be more than 90%. LC-ESI-MS/MS analysis showed the average theoretical mass of F2 to be 679.8 using m/z ratios. In conclusion, the findings suggest that F2 peptide is an effective inducer of apoptosis of cancer cells, thus offers an important strategy in the development of cancer therapeutics.
Subject(s)
Apoptosis/drug effects , Cell Cycle/drug effects , Cell Proliferation/drug effects , Chromatography, High Pressure Liquid , HeLa Cells , Humans , Lung Neoplasms/drug therapy , Lung Neoplasms/pathology , Peptide Fragments/isolation & purification , Peptide Fragments/pharmacology , Polyalthia/chemistry , Seeds/chemistry , Spectrometry, Mass, Electrospray Ionization , Spectroscopy, Fourier Transform Infrared , Tandem Mass Spectrometry , Tumor Cells, CulturedABSTRACT
<p><b>OBJECTIVE</b>To investigate the chemical constituents of the branches and leaves of Polyalthia nemoralis.</p><p><b>METHOD</b>The compounds were isolated and purified by silica gel, macroporous adsorption resin and Sephadex LH-20 column chromatographic methods. Their chemical structures were elucidated on the basis of physicochemical properties and spectral data.</p><p><b>RESULT</b>Fourteen compounds were isolated and identified as syringic acid (1), 3-methoxy-4-hydroxycinnamic acid (2), vanillic acid (3), 4-hydroxybenzoic acid (4), mauritianin (5), (+)-xylopinidine (6), (+)-oblongine(7), (+)-tembetarine (8), eythritol (9), D-mannitol (10), ethyl-beta-D-glucopyranoside (11), (+)-magnoflorine (12), stepharanine (13), (2S, 4R)-4-hydroxy-2-piperidine-carboxylic acid (14), respectively.</p><p><b>CONCLUSION</b>All the compounds were isolated from the genus Polyalthia for the first time; compounds 6 and 13 showed inhibitation activities against multi tumor cell lines.</p>
Subject(s)
Humans , Alkaloids , Chemistry , Pharmacology , Antineoplastic Agents, Phytogenic , Chemistry , Pharmacology , Aporphines , Chemistry , Pharmacology , Cell Line, Tumor , Cell Survival , Chromatography, Agarose , Methods , Coumaric Acids , Chemistry , Pharmacology , Gallic Acid , Chemistry , Pharmacology , Kaempferols , Chemistry , Pharmacology , Parabens , Chemistry , Pharmacology , Plant Extracts , Chemistry , Pharmacology , Plant Leaves , Chemistry , Plant Stems , Chemistry , Polyalthia , Chemistry , Vanillic Acid , Chemistry , PharmacologyABSTRACT
OBJECTIVE@#To explore antiulcer and antimicrobial properties of methanolic extract of Polyalthia longifolia var. pendula.@*METHODS@#Gastroprotective potential of Polyalthia longifolia was studied on ethanol and ethanol/HCl induced ulcers at 2 different doses (270 and 540 mg/kg/body weight). Antimicrobial efficacy of Polyalthia longifolia (25 mg/mL) was also studied against six gram positive, seven gram negative bacteria and five fungi by agar well diffusion method. Minimum inhibitory concentration was determined by agar well diffusion method in two fold serial dilution, in the range of 97-25,000 μg/mL.@*RESULTS@#The reduction of ulcer index in Polyalthia longifolia treated animals was found to be statistically significant with respect to control animals. The Polyalthia longifolia exhibited ulcer protection activity in dose dependent manner and was also better than the standard. In antimicrobial activity, gram positive bacteria were more susceptible to Polyalthia longifolia than gram negative bacteria and fungal strains.@*CONCLUSIONS@#Results obtained confirm the antiulcer and antimicrobial potential of the Polyalthia longifolia.
Subject(s)
Animals , Female , Humans , Male , Rats , Anti-Bacterial Agents , Anti-Infective Agents , Bacteria , Disease Models, Animal , Ethanol , Fungi , Hydrochloric Acid , Plant Extracts , Plant Leaves , Chemistry , Polyalthia , Chemistry , Protective Agents , Rats, Wistar , Ulcer , Drug Therapy , MicrobiologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the active constituents of the branches and leaves of Polyalthia nemoralis.</p><p><b>METHOD</b>The compounds were isolated and purified by silica gel, macroporous adsorption resin and Sephadex LH-20 column chromatographic methods. Their chemical structures were elucidated on the basis of physicochemical properties and spectral data.</p><p><b>RESULT</b>Eight compounds were isolated and identified as: zincpolyanemin (1), nickel bis-(pyridine-N-oxide-2-thiolate) (2), cupric bis (pyridine-N-oxide-2-thiolate (3), 2-methanesulfonyl-pyridine (4), 2-pyridinethiolate N-oxide (5), 2,2'-dithiodipyridine (6), 2-thiohydroxypyridin-N-oxide-2-S-beta-D-gluco pyranoside (7) and pyridine-N-oxide (8), respectively.</p><p><b>CONCLUSION</b>Compounds 2, 4-6, 8 were new natural products. The bioassays in vitro against five human tumor cell lines with MTT method showed stronger cytotoxic activities (IC50 0.05-0.09 mg x L(-1)) for compounds 1-3 and 6, and weaker cytotoxic activities (IC50 5.49-7.71 mg x L(-1)) for compound 5.</p>
Subject(s)
Humans , Cell Line, Tumor , Cell Survival , Drugs, Chinese Herbal , Chemistry , Pharmacology , Magnetic Resonance Spectroscopy , Plant Leaves , Chemistry , Plant Stems , Chemistry , Polyalthia , Chemistry , Pyridines , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To investigate the aporphine alkaloids in the branches and leaves of Polyalthia nemoralis.</p><p><b>METHOD</b>The compounds were isolated and purified by silica gel and Sephadex LH-20 column chromatographies. Their chemical structures were elucidated on the basis of physicochemical properties and spectral data.</p><p><b>RESULT</b>Five aporphine alkaloids were isolated and identified as: bidebiline A (1), annobraine (2), lanuginosine (3), liriodenine (4), oxostephanosine (5), respectively.</p><p><b>CONCLUSION</b>For the first time, Compounds 2 and 5 were obtained from Polyalthia while 1, 3 and 4 isolated from this plant. The bioassays in vitro against five human tumor cell lines with MTT method showed moderate cytotoxic activities (IC50 1 mg x L(-1)) of compounds 3-5.</p>