Insect antifeedants from tropical plants II: structure of zumsin

A novel A-seco limonoid was isolated from methanolic extract of Croton jatrophoides and designated as zumsin. This compound showed potent antifeedant activity against two lepidopteran larvae, pink bollworm, Pectinophora gossypiella (PC(50) = 1 microg/cm(2), PC(95) = 8 microg/cm(2)), and fall armyworm, Spodoptera frugiperda (PC(50) = 2 microg/cm(2), PC(95) = 16 microg/cm(2)). The structure of zumsin was determined as 1 using a variety of spectroscopic methods including nuclear magnetic resonance, mass spectrometry, and circular dichroism. The structure consists of an A'-B trans-fused ring while dumsin (2), a constituent of the same source, maintains an A'-B cis-fused ring, and suggests two unique biosynthetic processes after A ring oxidative expansion.     

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.     

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.     

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.     

Insecticidal sesquiterpene from Alpinia oxyphylla against Drosophila melanogaster

In the course of screening for novel naturally occurring insecticides from Chinese crude drugs, an MeOH extract of Alpinia oxyphylla was found to possess insecticidal activity against larvae of Drosophila melanogaster Meigen. From the extract, an insecticidal compound was isolated by bioassay-guided fractionation and identified as nootkatone (1) by GC, GC-MS, and (1)H and (13)C NMR spectroscopy. In bioassays for insecticidal activity, 1 showed an LC(50) value of 11.5 micromol/mL of diet against larvae of D. melanogaster and an LD(50) value of 96 microg/adult against adults. Epinootkatol (1A), however, showed slight insecticidal activity in both assays, indicating that the carbonyl group at the 2-position in 1 was the important function for enhanced activity of 1.     

Biologically active carbazole alkaloids from Murraya koenigii

The bioassay guided fractionation of the acetone extract of the fresh leaves of Murraya koenigii resulted in the isolation of three bioactive carbazole alkaloids, mahanimbine (1), murrayanol (2), and mahanine (3), as confirmed from their (1)H and (13)C NMR spectral data. Compound 2 showed an IC(50) of 109 microg/mL against hPGHS-1 and an IC(50) of 218 microg/mL against hPGHS-2 in antiinflammatory assays, while compound 1 displayed antioxidant activity at 33.1 microg/mL. All three compounds were mosquitocidal and antimicrobial and exhibited topoisomerase I and II inhibition activities.     

Phenolic constituents and antioxidant properties of Xanthosoma violaceum leaves

An extract of Xanthosoma violaceum leaves was subjected to a polyphenol profile determination, including total polyphenols, and antioxidant activity evaluation. Analysis of the extract resulted in the isolation of a new flavone C-glycoside, apigenin 6-C-beta-D-glucopyranosyl-8-C-beta-D-apiofuranoside (1), as well as known flavone C-glycosides, including vitexin (2), isovitexin (3), isovitexin 4'-O-rhamnopyranoside (4), apigenin 6-C-[beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranoside] (5), and apigenin 6,8-diC-beta-D-glucopyranoside (6). The antioxidant activity of the extract was assessed by means of two different in vitro tests: bleaching of the stable 1,1-diphenyl-2-picrylhydrazyl radical (DPPH test) and peroxidation induced by the water-soluble radical initiator 2,2'-azobis(2-amidinopropane) hydrochloride, on mixed dipalmitoylphosphatidylcholine/linoleic acid unilamellar vesicles (LP-LUV test). In both tests used, the extract and a fraction II showed a significant antioxidant/free-radical scavenging effect (fraction II, EC(50) = 11.6 microg/mL) in comparison to alpha-tocopherol (EC(50) = 10.1 microg/mL).     

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.     

Acaricidal activity of Paeonia suffruticosa root bark-derived compounds against Dermatophagoides farinae and Dermatophagoides pteronyssinus (Acari: Pyroglyphidae)

The acaricidal activities of materials derived from the root bark of Paeonia suffruticosa against adults of Dermatophagoides farinae and Dermatophagoides pteronyssinus were examined using direct contact and fumigation bioassays and compared with those of benzyl benzoate, dibutyl phthalate, and N,N-diethyl-m-toluamide (deet), widely used acaricides. The active constituents of Paeonia root bark were identified as paeonol and benzoic acid by spectroscopic analyses. On the basis of 24-h LD50 values, the acaricidal activities of paeonol (7.82 microg/cm3) and benzoic acid (6.58 microg/cm3) against adult D. farinae were comparable to that of benzyl benzoate (7.72 microg/cm3) but higher than those of deet (36.34 microg/cm3) and dibutyl phthalate (33.92 microg/cm3). Against adult D. pteronyssinus, the acaricidal activities of paeonol (7.08 microg/cm3) and benzyl benzoate (7.22 microg/cm3) were comparable to that of benzyl benzoate (7.14 microg/cm3). Deet and dibutyl phthalate were less effective. In fumigation tests with both mite species, paeonol and benzoic acid were much more effective in closed containers than open ones, indicating that the effect of these compounds was largely a result of action in the vapor phase. Neither benzyl benzoate, deet, nor dibutyl phthalate exhibited fumigant toxicity. Paeonia root bark-derived materials, particularly paeonol and benzoic acid, merit further study as potential acaricides or lead compounds for the control of D. farinae and D. pteronyssinus.     

Procyanidin oligomers from Japanese quince (Chaenomeles japonica) fruit inhibit activity of MMP-2 and MMP-9 metalloproteinases

The influence of procyanidin extract from Japanese quince fruit on the activities of matrix metalloproteinases MMP-2 and MMP-9 secreted to culture medium by human peripheral blood mononuclear cells (PBMC) and by human leukemia HL-60 cells was investigated by gelatin zymography. The extract proved to be an effective inhibitor of the enzymes activities (for MMP-2 and MMP-9 secreted by PBMC IC50 = 16-19 microg extract/mL and 22-25 microg extract/mL, respectively). To identify the most effective components of the extract it was fractionated by means of column chromatography on TSKgel Toyopearl HW-40 (S) bed. The obtained fractions were analyzed by TLC, HPLC, and MALDI-TOF MS. Their antioxidant activity was measured as cation radicals ABTS(.+) scavenging efficiency. The fractions VIII-XIV containing oligomers from trimer to hexamer (and probably higher oligomers) appeared to be the most effective inhibitors of MMP-2 and MMP-9 activity (IC50 value close to 4.6 microg total polyphenols/mL). To the best of our knowledge, it is the first report on gelatinase-inhibitory activity of Japanese quince fruit polyphenol extract. We conclude that polyphenols from Japanese quince can be used in cancer chemoprevention, although further studies are needed to elucidate the mechanisms underlying their biological activities.     

Insect antifeedants from tropical plants: structures of dumnin and dumsenin

Two novel A-seco limonoids, dumnin and dumsenin, were isolated from the methanolic extract of Croton jatrophoides by bioassay-guided fractionation, and the structures were determined by nuclear magnetic resonance, circular dichroism, and mass spectrometry experiments. These compounds showed potent antifeedant activity (PC(50) 相似文献   

Acaricidal activity of clove bud oil compounds against Dermatophagoides farinae and Dermatophagoides pteronyssinus (Acari: Pyroglyphidae)

The acaricidal activity of clove (Eugenia caryophyllata) bud oil-derived eugenol and its congeners (acetyleugenol, isoeugenol, and methyleugenol) against adults of Dermatophagoides farinae and Dermatophagoides pteronyssinus was examined using direct contact application and fumigation methods and compared with those of benzyl benzoate and N,N-diethyl-m-toluamide (DEET). Responses varied according to compound, dose, and mite species. On the basis of LD(50) values, the compound most toxic to D. farinae adults was methyleugenol (0.94 microg/cm(2)) followed by isoeugenol (5.17 microg/cm(2)), eugenol (5.47 microg/cm(2)), benzyl benzoate (9.22 microg/cm(2)), and acetyleugenol (14.16 microg/cm(2)). Very low activity was observed with DEET (37.59 microg/cm(2)). Against D. pteronyssinus adults, methyleugenol (0.67 microg/cm(2)) was much more effective than isoeugenol (1.55 microg/cm(2)), eugenol (3.71 microg/cm(2)), acetyleugenol (5.41 microg/cm(2)), and benzyl benzoate (6.59 microg/cm(2)). DEET (17.85 microg/cm(2)) was least toxic. These results indicate that the lipophilicity of the four phenylpropenes plays a crucial role in dust mite toxicity. The typical poisoning symptom of eugenol and its congeners was a similar death symptom of the forelegs extended forward together, leading to death without knockdown, whereas benzyl benzoate and DEET caused death following uncoordinated behavior. In a fumigation test with both mite species, all four phenylpropenes were much more effective in closed containers than in open ones, indicating that the mode of delivery of these compounds was largely due to action in the vapor phase. Eugenol and its congeners merit further study as potential house dust mite control agents or as lead compounds.     

A piperidine amide extracted from Piper longum L. fruit shows activity against Aedes aegypti mosquito larvae

Mosquito larvicidal activity of Piper longum fruit-derived materials against the fourth-instar larvae of Aedes aegypti was examined. A crude methanol extract of P. longum fruits was found to be active against the larvae, and the hexane fraction of the methanol extract showed a strong larvicidal activity of 100% mortality. The biologically active component of P. longum fruits was characterized as pipernonaline by spectroscopic analyses. The LC(50) value of pipernonaline was 0.25 mg/L. The toxicity of pipernonaline is comparable to that of pirimiphos-methyl as a mosquito larvicide. In tests with available components derived from P. longum, no activity was observed with piperettine, piperine, or piperlongumine.     

Antioxidant activity and characterization of volatile constituents of Taheebo (Tabebuia impetiginosa Martius ex DC)

Volatiles were isolated from the dried inner bark of Tabebuia impetiginosa using steam distillation under reduced pressure followed by continuous liquid-liquid extraction. The extract was analyzed by gas chromatography and gas chromatography-mass spectrometry. The major volatile constituents of T. impetiginosa were 4-methoxybenzaldehyde (52.84 microg/g), 4-methoxyphenol (38.91 microg/g), 5-allyl-1,2,3-trimethoxybenzene (elemicin; 34.15 microg/g), 1-methoxy-4-(1E)-1-propenylbenzene (trans-anethole; 33.75 microg/g), and 4-methoxybenzyl alcohol (30.29 microg/g). The antioxidant activity of the volatiles was evaluated using two different assays. The extract exhibited a potent inhibitory effect on the formation of conjugated diene hydroperoxides (from methyl linoleate) at a concentration of 1000 microg/mL. The extract also inhibited the oxidation of hexanal for 40 days at a level of 5 microg/mL. The antioxidative activity of T. impetiginosa volatiles was comparable with that of the well-known antioxidants, alpha-tocopherol, and butylated hydroxytoluene.     

Evaluation of insecticidal activity of diterpenes and lignans from Aristolochia malmeana against Anticarsia gemmatalis

The insecticidal activity of hexane extracts from the roots and leaves of Aristolochia malmeana was evaluated against Anticarsia gemmatalis larvae by topical application. Extract from the roots was the most active and caused 50% mortality in larvae at 308.4 microg/microL. From this extract, a clerodane diterpene, (-)-kolavenic acid, and three lignans, (-)-kusunokinin, (-)-hinokinin, and (8 S,8' R,9 S)-cubebin, were isolated by chromatography and partition procedures and then evaluated for their insecticidal activities either individually or in pairs. (-)-Kusunokinin showed higher activity against A. gemmatalis (LD10=9.3, LD50=230.1 microg/microL) than the crude extract, and its activity was dose-dependent, whereas the other constituents did not exhibit any significant activity. Together with (-)-kusunokinin and (-)-hinokinin, (-)-copalic acid, (-)-2-oxokolavenic acid, (-)- ent-6-beta-hydroxy-copalic acid, (8 R,8' R,9 R)- and (8 R,8' R,9 S)-cubebins, (-)-fargesin, and (-)-phillygenin were isolated from the hexane extract of the leaves. The compounds were identified on the basis of spectroscopic analysis.     

Acaricidal activity of butylidenephthalide identified in Cnidium officinale rhizome against dermatophagoides farinae and dermatophagoides pteronyssinus (Acari: Pyroglyphidae)

The acaricidal activity of materials derived from the rhizome of Cnidium officinale against adults of Dermatophagoides farinae and Dermatophagoides pteronyssinus was examined using direct contact application and fumigation methods and compared with that of benzyl benzoate and N,N-diethyl-m-toluamide (DEET). The active constituent of the Cnidium rhizome was identified as butylidenephthalide by spectroscopic analyses. Responses varied with dose. On the basis of 24-h LD(50) values, the acaricidal activity of butylidenephthalide (6.77 microg/cm(2)) against D. farinae adults was comparable to that of benzyl benzoate (8.54 microg/cm(2)). Very low activity was observed with DEET (37.59 microg/cm(2)). Against D. pteronyssinus adults, butylidenephthalide (6.46 microg/cm(2)) and benzyl benzoate (6.68 microg/cm(2)) were equitoxic. DEET (17.98 microg/cm(2)) was relatively inactive. The typical poisoning symptom of butylidenephthalide was lethargy of treated mites, leading to death without knockdown, whereas benzyl benzoate and DEET caused death following uncoordinated behavior. In a fumigation test with both mite species, butylidenephthalide was much more effective in closed containers than open ones. Naturally occurring C. officinale rhizome-derived materials merit further study as potential house dust mite control agents or lead compounds.     

Metabolites from Aspergillus fumigatus, an endophytic fungus associated with Melia azedarach, and their antifungal, antifeedant, and toxic activities

Thirty-nine fungal metabolites 1-39, including two new alkaloids, 12β-hydroxy-13α-methoxyverruculogen TR-2 (6) and 3-hydroxyfumiquinazoline A (16), were isolated from the fermentation broth of Aspergillus fumigatus LN-4, an endophytic fungus isolated from the stem bark of Melia azedarach. Their structures were elucidated on the basis of detailed spectroscopic analysis (mass spectrometry and one- and two-dimensional NMR experiments) and by comparison of their NMR data with those reported in the literature. These isolated compounds were evaluated for in vitro antifungal activities against some phytopathogenic fungi, toxicity against brine shrimps, and antifeedant activities against armyworm larvae (Mythimna separata Walker). Among them, sixteen compounds showed potent antifungal activities against phytopathogenic fungi (Botrytis cinerea, Alternaria solani, Alternaria alternata, Colletotrichum gloeosporioides, Fusarium solani, Fusarium oxysporum f. sp. niveum, Fusarium oxysporum f. sp. vasinfectum, and Gibberella saubinettii), and four of them, 12β-hydroxy-13α-methoxyverruculogen TR-2 (6), fumitremorgin B (7), verruculogen (8), and helvolic acid (39), exhibited antifungal activities with MIC values of 6.25-50 μg/mL, which were comparable to the two positive controls carbendazim and hymexazol. In addition, of eighteen that exerted moderate lethality toward brine shrimps, compounds 7 and 8 both showed significant toxicities with median lethal concentration (LC(50)) values of 13.6 and 15.8 μg/mL, respectively. Furthermore, among nine metabolites that were found to possess antifeedant activity against armyworm larvae, compounds 7 and 8 gave the best activity with antifeedant indexes (AFI) of 50.0% and 55.0%, respectively. Structure-activity relationships of the metabolites were also discussed.     

Effects of grape cell culture extracts on human topoisomerase II catalytic activity and characterization of active fractions

Grape and its cell culture extracts are rich in flavonoids and stilbenes that are biologically active. The objective of this study was to evaluate possible inhibitory effects of grape (a Vitis hybrid Bailey Alicant A) cell culture extract and subfractions on human DNA topoisomerase II catalytic activity and to characterize constituents in the most potent fractions. At 5 microg/mL, grape cell crude extract and Toyopearl (TP) fractions 2-6 provided significantly greater inhibition of topoisomerase II catalytic activity than quercetin, a chemopreventive agent previously known as a topoisomerase catalytic inhibitor. The most potent topoisomerase II catalytic inhibitors from grape cell culture extracts in descending order of potency were TP fractions 4 and 6 (IC(50) = 0.28-0.29 microg/mL), TP-3 (IC(50) = 0.74 microg/mL), and crude extract (IC(50) = 1.02 microg/mL); each was significantly more potent than resveratrol (IC(50) = 18.0 microg/mL), another well-known chemopreventive topoisomerase II catalytic inhibitor. Using both high-performance liquid chromatography and liquid chromatography electrospray ionization mass spectrometry, constituents in TP-4 and TP-6 were characterized. These constituents included cyanidin-3,5-diglucoside, malvidin-3-acetylglucoside, peonidin-3-coumaryl-5-diglucoside, procyanidin B(1), procyanidin B(2), procyanidin B(5), procyanidin dimer digallate, procyanidin C(1), myricetin, and rutin, none of which have been previously characterized from grape cell cultures. The significant potency especially of TP-4 and TP-6 from grape cell cultures suggests that these fractions may have potential as chemopreventive agents.     

Estrogenic activity of glycitein, a soy isoflavone

Glycitein (4',7-dihydroxy-6-methoxyisoflavone) accounts for 5-10% of the total isoflavones in soy food products. The biological activity of this compound has not been reported to date, although numerous studies have been performed with the other soy isoflavones, daidzein and genistein. Glycitein was isolated from soy germ to 99% purity. Weaning female B6D2F1 mice were dosed with glycitein (3 mg/day), genistein (3 mg/day), and diethylstilbestrol (DES) (0.03 microg/day) in 5% Tween 80 by gavage for 4 days. A control group received an equal volume of 5% Tween 80 solution daily. The uterine weight increased 150% with glycitein (p < 0.001), 50% with genistein (p < 0. 001), and 60% with DES (p < 0.001) compared with the control group. DES, 17beta-estradiol, and three isoflavones (daidzein, genistein, and glycitein) were examined for their competitive binding abilities with 17beta-((3)H)estradiol to the estrogen receptor proteins of the B6D2F1 mouse uterine cytosol. The concentrations of each compound required to displace 50% of the ((3)H)estradiol at 5 nM in the competitive binding assay were 1.15 nM DES, 1.09 nM 17beta-estradiol, 0.22 microM genistein, 4.00 microM daidzein, and 3.94 microM glycitein. These data indicated that glycitein has weak estrogenic activity, comparable to that of the other soy isoflavones but much lower than that of DES and 17beta-estradiol.     

Ovicidal and adulticidal effects of Eugenia caryophyllata bud and leaf oil compounds on Pediculus capitis

The toxicity of Eugenia caryophyllata bud and leaf oil-derived compounds (acetyleugenol, beta-caryophyllene, eugenol, alpha-humulene, and methyl salicylate) and congeners of eugenol (isoeugenol and methyleugenol) against eggs and females of Pediculus capitis was examined using direct contact application and fumigation methods and compared with those of the widely used delta-phenothrin and pyrethrum. In a filter paper diffusion bioassay with female P. capitis, the pediculicidal activity of the Eugenia bud and leaf oils was comparable to those of delta-phenothrin and pyrethrum on the basis of LT(50) values at 0.25 mg/cm(2). At 0.25 mg/cm(2), the compound most toxic to female P. capitis was eugenol followed by methyl salicylate. Acetyleugenol, beta-caryophyllene, alpha-humulene, isoeugenol, and methyleugenol were not effective. Eugenol at 0.25 mg/cm(2) was as potent as delta-phenothrin and pyrethrum but was slightly less effective than the pyrethroids at 0.125 mg/cm(2). Against P. capitis eggs, methyl salicylate and eugenol were highly effective at 0.25 and 1.0 mg/cm(2), respectively, whereas little or no activity at 5 mg/cm(2) was observed with the other test compounds as well as with delta-phenothrin and pyrethrum. In fumigation tests with female P. capitis at 0.25 mg/cm(2), eugenol and methyl salicylate were more effective in closed cups than in open ones, indicating that the effect of the compounds was largely due to action in the vapor phase. Neither delta-phenothrin nor pyrethrum exhibited fumigant toxicity. The Eugenia bud and leaf essential oils, particularly eugenol and methyl salicylate, merit further study as potential P. capitis control agents or lead compounds.