Biologically active components against Drosophila melanogaster from Podophyllum hexandrum

In the course of screening for novel naturally occurring insecticides from Chinese crude drugs, a dichloromethane extract of Podophyllum hexandrum was found to give an insecticidal activity against larvae of Drosophila melanogaster Meigen. From the extract, an insecticidal compound was isolated by bioassay-guided fractionation. The compound was identified as podophyllotoxin (1) by comparison of its spectroscopic characteristics with literature data. In bioassays for insecticidal activity, 1 showed a LC(50) value of 0.24 micromol/mL diet against larvae of D. melanogaster and a LD(50) value of 22 microg/adult against adults. Acetylpodophyllotoxin (1A), however showed slight insecticidal activity in both assays, indicating that the 4-hydroxyl group was an important function for enhanced activity of 1.     

Phenolic antioxidants from the heartwood of Acacia confusa

In the present study, the ethanolic extracts from the heartwood of Acacia confusa, a species indigenous to Taiwan, exhibit strong antioxidant effects. Among all the fractions from ethanolic extracts of heartwood, the EtOAc soluble fraction exhibits the best antioxidant activity. The 80% 1,1-diphenyl-2-picrylhydrazyl radical inhibitory activity by the EtOAc extract was observed at a concentration of 5 mug/mL, and at the same dosage there was a similar free radical scavenging activity for (-)-ascorbic acid and (+)-catechin, both of which are well-known antioxidants. In addition, the EtOAc extract also protects PhiX174 supercoiled DNA against strand scission induced by hydroxyl radical. Furthermore, following by column chromatography and reverse-phase high-performance liquid chromatorgrapy, 10 pure phenolic compounds, including three major antioxidants (3,7,8,3',4'-pentahydroxyflavone, 7,8,3',4'-tetrahydroxy-3-methoxyflavone, and 3,4,2',3',4'-pentahydroxy-trans-chalcone) and a new flavonoid (3,7,8,3'-tetrahydroxy-4'-methoxyflavone), were isolated from the ethanolic extracts of A. confusa heartwood.     

Insecticidal effect of phthalides and furanocoumarins from Angelica acutiloba against Drosophila melanogaster

Insecticidal activity of Angelica acutiloba extract and its constituents was investigated and compared with that of rotenone. Bioassay-guided isolation of the chloroform extract of A. acutiloba against larvae of Drosophila melanogaster afforded two phthalides, (Z)-butylidenephthalide (1) and (Z)-ligustilide (2), and two furanocoumarins, xanthotoxin (3) and isopimpinellin (4). The structures of these compounds were established by spectroscopic analysis. The isolated compounds 1, 2, 3, and 4 exhibited LC(50) values of 0.94, 2.54, 3.35, and 0.82 micromol/mL of diet concentration against larvae of D. melanogaster, respectively. Against both sexes (males/females, 1:1) of adults (5-7 days old), compound 1 showed the most potent activity with a LD(50) value of 0.84 microg/adult. Compound 1 is a more active insecticide than rotenone (LD(50) = 3.68 microg/adult) and has potential as a novel insect control agent. However, compound 2 was inactive against adults. The structure-activity relationship of phthalides isolated indicated that the aromaticity appeared to play an important role in the activity of both larvae and adults. To determine the insecticide mode of action for acute adulticidal activity, acetylcholinesterase (AChE) inhibitory activity was also investigated in vitro, and the result indicated that the acute adulticidal activity of compounds 3 and 4 was due to the inhibition of AChE.     

Sesamolinol glucoside, disaminyl ether, and other lignans from sesame seeds

The application of a procedure based on XAD-4 adsorption resin permitted the obtainment of an enriched polyphenolic extract from Sesamum indicum seeds. Chemical analysis of the obtained extract led to the identification of 12 lignans. Among them, 2 lignans, (+)-sesamolinol-4'-O-β-D-glucoside and disaminyl ether, are reported for the first time as natural compounds. Their structure has been determined by spectroscopic methods, mainly by the application of one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) techniques [heteronuclear multiple-quantum coherence (HMQC), heteronuclear multiple-bond correlation (HMBC), and nuclear Overhauser effect spectrometry (NOESY)] and mass spectroscopy. The isolated compounds were evaluated for their antimutagenic activity. Among the tested lignans, the most active lignan was found to be sesamolin, followed by sesamolinol and samin, against H(2)O(2). Additionally, some of the tested lignans showed desmutagenic activity against benzo[a]pyrene (BaP).     

Tyrosinase inhibitor from black rice bran

The inhibitor of tyrosinase activity in black rice bran was investigated. The methanol extract from black rice bran was re-extracted with hexane, chloroform, ethyl acetate, or water. The ethyl acetate extract had the most potent inhibition against tyrosinase activity by 80.5% at a concentration of 0.4 mg/mL. Inhibitory compound in the ethyl acetate fraction was isolated by silica gel column chromatography, and identified as protocatechuic acid methyl ester (compound 1) by GC, GC-MS, IR, and 1H and 13C NMR spectroscopy. Compound 1 inhibited 75.4% of tyrosinase activity at a concentration of 0.50 micromol/mL. ID(50) (50% inhibition dose) value of compound 1 was 0.28 micromol/mL. To study the structure-activity relationship, protocatechuic acid (2), vanillic acid (3), vanillic acid methyl ester (4), isovanillic acid (5), isovanillic acid methyl ester (6), veratric acid (7), and veratric acid methyl ester (8) were also assayed.     

Glabrene and isoliquiritigenin as tyrosinase inhibitors from licorice roots

Tyrosinase is known to be a key enzyme in melanin biosynthesis, involved in determining the color of mammalian skin and hair. Various dermatological disorders, such as melasama, age spots, and sites of actinic damage, arise from the accumulation of an excessive level of epidermal pigmentation. The inadequacy of current therapies to treat these conditions as well as high cytotoxicity and mutagenicity, poor skin penetration, and low stability of formulations led us to seek new whitening agents to meet the medical requirements for depigmenting agents. The inhibitory effect of licorice extract on tyrosinase activity was higher than that expected from the level of glabridin in the extract. This led us to test for other components that may contribute to this strong inhibitory activity. Results indicated that glabrene and isoliquiritigenin (2',4',4-trihydroxychalcone) in the licorice extract can inhibit both mono- and diphenolase tyrosinase activities. The IC(50) values for glabrene and isoliquiritigenin were 3.5 and 8.1 microM, respectively, when tyrosine was used as substrate. The effects of glabrene and isoliquiritigenin on tyrosinase activity were dose-dependent and correlated to their ability to inhibit melanin formation in melanocytes. This is the first study indicating that glabrene and isoliquiritigenin exert varying degrees of inhibition on tyrosinase-dependent melanin biosynthesis, suggesting that isoflavenes and chalcones may serve as candidates for skin-lightening agents.     

Antifeedant and mosquitocidal compounds from Delphinium x cultorum cv. Magic fountains flowers

Six volatile compounds, ethylmethylbenzene (1), 1-isopentyl-2,4, 5-trimethylbenzene (2), 2-(hex-3-ene-2-one)phenylmethyl ketone (3), E and Z isomers of 3-butylidene-3H-isobenzofuran-1-one (4 and 5), and 2-penten-1-ylbenzoic acid (6), were isolated from the mosquitocidal hexane extract of Delphinium x cultorum cv. Magic Fountains flowers. In addition, the ethyl acetate extract, which displayed corn earworm antifeedant activity, yielded 4-hydroxybenzoic acid (7) and bis(4-hydroxyphenyl)methanol (8). However, compounds 7 and 8 were not biologically active.     

Action mechanism of an angiotensin I-converting enzyme inhibitory peptide derived from chicken breast muscle

In a previous study, we isolated the inhibitory peptide (P4 peptide, Gly-Phe-Hyp-Gly-Thr-Hyp-Gly-Leu-Hyp-Gly-Phe) for angiotensin I-converting enzyme (ACE) from chicken breast muscle extract possessing hypotensive activity for spontaneously hypertensive rats (SHRs). This study was performed to elucidate the peptide's action mechanisms of inhibiting ACE. Intravenous administration of synthetic P4 peptide resulted in significant drops in the blood pressures of SHRs. As Dixon plots indicate, the P4 peptide showed high affinity toward ACE (K(i) = 11.48 microM) and only 10% of the total amount of the P4 peptide was decomposed. The analyses of the relationship between the ACE inhibitory activity and structure of the P4 peptide clarified that Hyp-Gly-Leu-Hyp-Gly-Phe showed a stronger activity (IC50 = 10 microM) than the P4 peptide (IC50 = 46 microM). When Phe at the C-terminus of the P4 peptide was deleted, IC50 changed to 25000 microM, indicating that Phe at the C-terminus of the peptide is very important for ACE inhibitory activity.     

Lupane-type saponins from leaves of Acanthopanax sessiliflorus and their inhibitory activity on pancreatic lipase

Three known saponins, chiisanoside, 11-deoxyisochiisanoside, and isochiisanoside, and one novel saponin, 3,4-seco-4(23),20(29)-lupadiene-3,28-dioic acid 28-O-alpha-l-rhamnopyranosyl (1-->4)-beta-d-glucopyranosyl (1-->6)-beta-d-glucopyranoside, referred to as sessiloside, were isolated from a hot water extract of Acanthopanax sessiliflorus leaves. All of these saponins were lupane-type triterpene triglycosides, and their concentrations were 4.1, 1.0, 0.5, and 0.4% (w/w) of the total extract, respectively. Sessiloside and chiisanoside inhibited pancreatic lipase activity in vitro, and addition of the saponin-rich fraction to a high-fat diet suppressed the body weight gain of mice. The possibility of application of the lupane-type saponins from A. sessiliflorus leaves to the treatment of obesity is discussed.     

Antifungal effects of different organic extracts from Melia azedarach L. on phytopathogenic fungi and their isolated active components

Extracts from different parts of Melia azedarach L. were studied as potential antifungal agents for selected phytopathogenic fungi. In a serial agar dilution method, hexanic and ethanolic extracts from fruit, seed kernels, and senescent leaves exhibited fungistatic activity against Aspergillus flavus,Diaporthe phaseolorum var. meridionales, Fusarium oxysporum, Fusarium solani, Fusarium verticillioides, and Sclerotinia sclerotiorum. Both hexanic extract from senescent leaves and ethanolic extract from seed kernel were highly effective on all tested fungi, with minimum inhibitory concentration (MIC) values ranging from 0.5 to 25 mg/mL and 0.5 to 5 mg/mL, respectively. In addition, all of the above-mentioned extracts showed fungicidal activity on these fungi, with ethanolic seed kernel extract being the most active. Three compounds displaying activity against F. verticillioides were isolated from the ethanolic seed kernel extract and were characterized as vanillin (1), 4-hydroxy-3-methoxycinnamaldehyde (2), and (+/-)-pinoresinol (3), with MICs of 0.6, 0.4, and 1.0 mg/mL, respectively. These compounds also showed a synergistic effect when combined in different concentrations, needing four times less concentration to reach complete inhibition in the growth of F. verticillioides.     

Antioxidative activity of carbazoles from Murraya koenigii leaves.

The antioxidative properties of the leaves extracts of Murraya koenigii using different solvents were evaluated based on the oil stability index (OSI) together with their radical scavenging ability against 1-1-diphenyl-2-picrylhydrazyl (DPPH). The methylene chloride (CH(2)Cl(2)) extract and the ethyl acetate (EtOAc) soluble fraction of the 70% acetone extract significantly prolonged the OSI values comparable to those of alpha-tocopherol and BHT. Five carbazole alkaloids were isolated from the CH(2)Cl(2) extract and their structures were identified to be euchrestine B (1), bismurrayafoline E (2), mahanine (3), mahanimbicine (4), and mahanimbine (5) based on (1)H and (13)C NMR and mass (MS) spectral data. The OSI value of carbazoles at 110 degrees C decreased in the order 1 and 3 > alpha-tocopherol > BHT > 2 > 4, 5 and control. It is assumed that compounds 1 and 3 contributed to the high OSI value of the CH(2)Cl(2) extract of M. koenigii. The DPPH radical scavenging activity for these carbazoles was in the order ascorbic acid > 2 > 1, 3 and alpha-tocopherol > BHT > 4 and 5.     

Insect antifeedants from Munronia henryi: structure of munroniamide

A novel A,B-seco-tetranortriterpenoid lactam, named munroniamide (1), along with three known ceramides (2-4), was isolated from the methanolic extract of the whole bodies of Munronia henryi. The structure of 1 was established using spectroscopic methods. Compound 2 exhibited significant antifeeding activity, and 1 exhibited moderate activity, whereas 3 and 4 showed negative activity against Pieris brassicae L.     

Potential food additives from Carex distachya roots: identification and in vitro antioxidant properties

In the present study, the chemical composition and antioxidant properties of root methanol extract of Carex distachya Desf. (Cyperaceae) were assessed to use this plant as sources of food additives and nutraceuticals. The IC50 of the extract (4.2 microg/mL), derived from the DPPH radical scavenging capacity assay, was similar to those of ascorbic acid, alpha-tocopherol, and BHT. These results revealed a strong antioxidant activity because of the presence of an extraordinary quantity of bioactive phytochemicals. The phytochemical study of the root extract led to the isolation and identification of new and known polyphenols, most of them common constituents of plant foods. A total of 16 polyphenols, identified on the basis of spectroscopic data as 7 lignans, 4 phenylethanoids, 3 resveratrol derivatives, a monolignol, and a secoiridoid glucoside, were isolated. The tentative structural elucidation of the new metabolites 5'-O-beta-D-glucopyranosyloxy-3,3'-dimethoxy-7,9'-epoxylignan-4,8',9-triol and 3,5-bis-O-beta-D-glucopyranosyloxy-3'-methoxy-trans-stilben-4'-ol have been performed by a combined approach using ESI/TQ/MS techniques and 1D and 2D NMR experiments. All of the compounds have been tested for their antioxidant activity using six different antioxidant and radical scavenging tests. Interestingly, the extract contained high quantities of polyphenols, most of them reported as constituents of edible plants, such as grape and olive, suggesting that the methanol root extract of this plant could be used as a source of natural antioxidants useful as potential food additives.     

Larvicidal and adulticidal activity of alkylphthalide derivatives from rhizome of Cnidium officinale against Drosophila melanogaster

The insecticidal activity of the chloroform extract of Cnidium officinale rhizomes and its constituents was investigated against larvae and adults of Drosophila melanogaster and compared with that of rotenone. Bioassay-guided isolation of the chloroform extract of C. officinale resulted in the isolation and characterization of four alkylphthalides, cnidilide (1), (Z)-ligustilide (2), (3S)-butylphthalide (3), and neocnidilide (4). The structures of these compounds were established by spectroscopic analysis. The isolated compounds 2, 3, and 4 exhibited LC50 values of 2.54, 4.99, and 9.90 micromol/mL of diet concentration against larvae of D. melanogaster, respectively. Against both sexes (males/females, 1:1) of adults (5-7 days old), compound 3 showed the most potent activity of the compounds isolated with the LD50 value of 5.93 microg/adult, comparable to that of rotenone (LD50 = 3.68 microg/adult). Structure-activity relationships of phthalides isolated suggest that the presence of conjugation with the carbonyl group in the lactone ring appeared to play an important role in the larvicidal activity. Acetylcholinesterase (prepared from the adult heads of D. melanogaster) inhibitory activity was also investigated in vitro to determine the insecticide mode of action for the acute adulticidal activity.     

Melanin synthesis inhibitors from Balanophora fungosa

Tyrosinase, trypsin, and tryptase are known to play important roles in melanin production of human skin. This paper describes the study of the inhibitory effect of Balanophora fungosa on melanin. The 50% EtOH extract obtained from B. fungosa indicated an inhibitory effect on mushroom tyrosinase activity with an IC(50) value of 15 μg/mL. Bioassay-guided fractionation of the active extract resulted in the isolation of four known compounds. Their structures were identified as 1-O-(E)-caffeoyl-3-O-galloyl-4,6-(S)-HHDP-β-d-glucopyranose (1), 1-O-(E)-caffeoyl-3,4,6-tri-O-galloyl-β-d-glucopyranose (2), caffeoyl-β-d-glucopyranose (3), and abietin (4) on the basis of spectroscopic analyses and comparison of their spectral data with those in the literature. Compounds 1 and 2 prevented pigmentation of melanin in a three-dimensional cultured human skin model. Furthermore, compounds 1 and 2 indicated inhibitory activities against trypsin and tryptase.     

Antifeedant and insecticide properties of a limonoid from Melia azedarach (Meliaceae) with potential use for pest management

In the course of screening for novel naturally occurring insecticides from plants, the activity of the fruit extract of the Argentinian Melia azedarach L. (Meliaceae) and its recently described limonoid meliartenin were investigated. The antifeedant activity of the fruit extract was tested on a variety of herbivore and granivorous insects through choice tests. Sixteen of 17 species belonging to three orders consume significantly less food when treated with the extract. The bioactivity of the isolated active compound meliartenin and its interchangeable isomer 12-hydroxiamoorastatin (1) was further studied. In choice tests, compound 1 inhibited feeding of Epilachna paenulata Germ. (Coleoptera, Coccinellidae) larvae, with an ED(50) value of 0.80 microg/cm(2), comparable to that of azadirachtin (2) and lower than that of toosendanin (3) (0.72 and 3.69 microg/cm(2), respectively), both compounds used for comparison purposes. In no-choice tests, E. paenulata larvae reared on food treated with 1 or 2 ate less, gained less weight, and suffered greater mortality rates than control larvae. The activity of compound 1 was comparable to that of 2, with LD(50) values of 0.76 and 1.24 microg/cm(2), respectively, at 96 h. Shorter LT(50) values were recorded for 1 at 4 and 1 microg/cm(2) in comparison with 2. Thus, M. azedarach fruit extract and its active principle have interesting potential for use in pest control programs.     

Antimutagenic activity of isoflavone from Pueraria lobata

A methanol extract from Pueraria lobata showed a suppressive effect on umu gene expression of the SOS response in Salmonella typhimurium TA1535/pSK1002 against the mutagen 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (furylfuramide). The methanol extract from P. lobata was re-extracted with hexane, dichloromethane, ethyl acetate, butanol, and water, respectively. A suppressive compound in the dichloromethane and ethyl acetate extract fractions was isolated by SiO(2) column chromatography and identified as tectorigenin (1) by EI-MS and (1)H and (13)C NMR spectroscopy. Compound 1 and its methylated derivative [7,4'-di-O-methyltectorigenin (2)] had the suppressive effects on umu gene expression of the SOS response in Salmonella typhimurium TA1535/pSK1002 against furylfuramide, 4-nitroquinoline-1-oxide, N-methyl-N'-nitrosoguanidine, and activated Trp-P-1, which do not require live metabolic activation by S9. These compounds also showed suppression of SOS-inducing activity against Trp-P-1 and AfB(1), which requires liver metabolizing enzymes. In addition to the antimutagenic activities of these compounds against furylfuramide, Trp-P-1 and activated Trp-P-1 were also assayed by an Ames test using S. typhimurium TA100.     

Antifungal activity of thiophenes from Echinops ritro

Extracts from 30 plants of the Greek flora were evaluated for their antifungal activity using direct bioautography assays with three Colletotrichum species. Among the bioactive extracts, the dichloromethane extract of the radix of Echinops ritro (Asteraceae) was the most potent. Bioassay-guided fractionation of this extract led to the isolation of eight thiophenes. Antifungal activities of isolated compounds together with a previously isolated thiophene from Echinops transiliensis were first evaluated by bioautography and subsequently evaluated in greater detail using a broth microdilution assay against plant pathogens Colletotrichum acutatum, Colletotrichum fragariae, Colletotrichum gloeosporioides, Botrytis cinerea, Fusarium oxysporum, Phomopsis viticola, and Phomopsis obscurans. 5'-(3-Buten-1-ynyl)-2,2'-bithiophen (1), alpha-terthienyl (2), and 2-[pent-1,3-diynyl]-5-[4-hydroxybut-1-ynyl]thiophene (5) at 3 and 30 microM were active against all three Colletotrichum species, F. oxysporum, P. viticola, and P. obscurans.     

Insecticidal fatty acids and triglycerides from Dirca palustris.

Five compounds, 1-5, were isolated from the seed hexane extract of Dirca palustris. Compounds 1-3 were triglycerides, and 4 and 5 were linoleic and oleic acids, respectively. Compounds 1-3 were not biologically active; however, 4 (linoleic acid) and 5 (oleic acid) were insecticidal against fourth instar Aedes aegyptii larvae and exhibited potent feeding deterrent activity against neonate larvae of Helicoverpa zea, Lymantria dispar, Orgyia leucostigma, and Malacosoma disstria.     

Isolation and identification of antiplatelet aggregatory principles from the leaves of Piper lolot

The methanolic extract of Piper lolot, having shown potent inhibitory activity on platelet aggregation induced by arachidonic acid (AA) and platelet activating factor (PAF), was subjected to activity-guided isolation to yield twelve new amide alkaloids, piperlotine A-L (1-12), along with twenty-nine known compounds. Their structures were elucidated on the basis of spectroscopic analysis. The isolated compounds were tested for their inhibitory activity on the rabbit platelet aggregation. The compounds piperlotine A (1), piperlotine C (3), piperlotine D (4), piperlotine E (5), 3-phenyl-1-(2,4,6-trihydroxyphenyl)propan-1-one (21), 3-(4-methoxyphenyl)-1-(2,4,6-trihydroxyphenyl)propan-1-one (22), 1-trans-cinnamoylpyrrolidine (24), sarmentine (26), pellitorine (27), methyl 3-phenylpropionate (32), and (10S)-10-hydroxypheophorbide a methyl ester (40) showed potent antiplatelet aggregation activity.