Natural Product Communications

Online ISSN: 1555-9475
Print ISSN: 1934-578X
Cellulase is a collective term that encompasses enzymes which catalyze reactions that participate in the degradation of insoluble cellulose to soluble carbohydrates. In the present study, production of extra cellular cellulases by a filamentous fungus, Aspergillus niger FCBP-02, was studied in solid-state fermentation (SSF) as well as in submerged fermentation (SmF). Trials were conducted to evaluate the effect of mutagenesis by UV irradiation (5-40 min) and ethyl methane sulfonate (EMS) treatment (50-300 microg mL(-1)) to obtain hyper active cellulase enzyme producers among the potential strains. The enzyme activity assays of parental and mutant strains clearly revealed significantly higher cellulase activity of mutant A-Ch-5.5 (96 Units mL(-1)), followed by A-UV-5.6 (71 Units mL(-1)) with respect to the wild strain of A. niger FCBP-02 (53.7 Units mL(-1)). The profile of genetic variability among wild and mutant derivatives was scrutinized through RAPD-PCR. The expression pattern of mutants exhibited that the mutants were isogenic variants of the wild type and the out performance of the mutants could be attributed to the change in genetic make up.
The dioxo-lignans of the arylnaphthalene-type named justicidone (2) and elenodione (3) were obtained from elenoside (1) through a short and efficient semisynthetic process. Justicidone (2), one of its synthetic precursors, 4-(benzo[d][1,3]dioxol-5-yl)-5,6,8-trimethoxy-3a,4-dihydronaphtho[2,3-c]furan-1(3H)-one (9), and the aglycone of elenoside (5) showed cytotoxic activity towards the HL-60 cell line (IC50 = 7.25 miroM, 5.41 microM and 2.06 miroM, respectively).
The five-step synthesis of the new 4H-anthra[1,2-b]pyran derivative 1 is reported. The key steps in this approach included a Marschalk alkylation of 1,4-dihydroxyanthraquinone followed by a Baker-Venkataraman reaction and then an acid-catalyzed cyclization of ring A to form the 4H-anthra[1,2-b]pyran system. Two compounds, the 4H-anthra[1,2-b]pyran 1 and the anthraquinone derivative 6 were evaluated for antimicrobial activity and showed moderate antialgal, antifungal, and antibacterial activities.
Pyrogallol and its derivatives are biologically active compounds, and pyrogallol also forms the basis of an increasingly important tetrameric supramolecular scaffold. Pyrogallol[4]arenes are tetrameric macrocycles that form from 1,2,3-trihydroxybenzene and aldehydes under acidic conditions. Pyrogallol was treated with two equivalents of pivaloyl chloride to form pyrogallyl dipivaloate. A mixture of regioisomers was invariably obtained and a rapid equilibrium was observed between the 1,2- and 1,3-diesters in polar solvents. A pure sample of solid pyrogallyl 1,2-dipivaloate was isolated and its crystal structure was obtained. The pure compound was shown to rearrange to mixtures similar to those isolated initially.
The structure of a new secondary metabolite from Streptomyces sp. was determined as 4-acetyl-1,3-dihydroimidazo[4,5-c]pyridin-2-one by synthesis of the natural product itself and of the regioisomeric 7-acetylimidazo[4,5-b]pyridine derivative. The former compound was prepared, in 28% overall yield, in a sequence of nitration, reduction, condensation, and Stille reaction of 4-aminopyridine, while the regioisomer was obtained in 5% overall yield by amination, nitration, reduction, condensation, and oxidation of 4-ethylpyridine.
The enantioselective synthesis of (S)-1-hydroxy-1,3,5-bisabolatrien-10-one 1 is here described. This sesquiterpene was prepared using (S)-3-(2-methoxy-4-methyl-phenyl)butan-1-ol as a chiral building block. Two different pathways were employed and both turned out to be high yielding, affording 1 in good chemical purity and without any racemization of the existing stereocenter. The spectroscopic data of the synthetic (S)-1 were in very good agreement with those reported for the natural compound, which was extracted from Juniperus formosana heartwood and from the leaves of J. chinensis. The positive sign of the measured optical rotation value of synthetic (S)-1 allows the unambiguous assignment of the absolute configuration of (+)-1 as the (S)-enantiomer. This finding corrects the previous configuration determination which indicated the opposite result. At last, since even (R)-3-(2-methoxy-4-methyl-phenyl)butan-1-ol is preparable in high enantiomer purity by mean of a different biocatalytic process, the formal synthesis of natural (R)-1 was also accomplished.
2-Acetyl-1-hydroxynaphthalene was converted into the title compound in three steps (bromination, substitution and methylation). 1-Methoxynaphthalene on bromination, substitution and acetylation, respectively, also yielded the target compound.
Effect of 1,5-AF on PPARγ, C/EBPα and C/EBPβ expression in cultured 3T3-L1 preadipocytes. 3T3-L1 preadipocytes were cultured with a 0-800 μg/mL solution of 1,5-AF for 2 days following initiation of differentiation. PPARγ (A), C/EBPα (B) and C/EBPβ (C) protein levels were detected by Western blot. The PPARγ, C/EBPα and C/EBPβ expression levels are represented relative to control levels. The results are representative of three separate experiments. As shown in Figures 3A and 3B, control cells accumulated lipid droplets. However, mature adipocytes treated with 1,5-AF accumulated smaller lipid droplets than control cells, indicating that 1,5-AF also inhibits the accumulation of intracellular triglyceride in mature 3T3-L1 adipocytes. As shown in Figures 4A and 4B, 1,5-AF treatment reduced the protein levels of major adipogenic transcription factors, including PPARγ, and C/EBPα in mature adipocytes.
Effect of 1,5-AF on PPARγ and C/EBPα expression in cultured 3T3-L1 mature adipocytes. Fully differentiated 3T3-L1 cells were treated with a 0-800 μg/mL solution of 1,5-AF for 2 days. PPARγ (A) and C/EBPα (B) protein levels were detected by Western blot. The PPARγ and C/EBPα expression levels are represented relative to control levels. The results are representative of three separate experiments.  
Effect of 1,5-AF on intracellular ROS levels and AMPK phosphorylation in cultured 3T3-L1 preadipocytes. (A) Intracellular ROS levels. The 3T3-L1 preadipocytes were cultured for 3 h after initiation of differentiation with (a) 0 or (b) 800 μg/mL 1,5-AF. Phosphorylated AMPK (B) and AMPK (C) protein levels were detected by Western blot. 3T3-L1 preadipocytes were cultured for 2 days after initiation of differentiation with a 0-800 μg/mL solution 1,5-AF. The results are representative of three separate experiments.
Effect of 1,5-AF on Oil Red O staining and GPDH activity in cultured 3T3- L1 mature adipocytes. Fully differentiated 3T3-L1 cells were treated with a 0-800 μg/mL solution of 1,5-AF for 3 days. (A) Photograph of Oil Red O stained cells. (a) 0 μg/mL (Control), (b) 400 μg/mL and (c) 800 μg/mL 1,5-AF. (B) Lipid accumulation in cultured 3T3-L1 mature adipocytes. (C) GPDH activities in cultured 3T3-L1 mature adipocytes. The results are presented as the mean ± SD of four experiments. *p<0.05, **p<0.01 versus control cells.  
1,5-Anhydro-D-fructose (1,5-AF) is a monosaccharide that shares a structural similarity to glucose. 1,5-AF is found in fungi, algae, Escherichia coli and rat liver and is produced by the degradation of starch and glycogen, which is catalyzed by the enzyme alpha-1,4-glucan lyase. However, the physiological role of 1,5-AF in mammalian tissues is not well understood. Here, we investigated the anti-obesity potential of 1,5-AF on adipogenesis in 3T3-L1 adipocytes. 1,5-AF caused a significant decrease in GPDH activity in 3T3-L1 preadipocytes and mature adipocytes without eliciting cytotoxicity, and inhibited cellular lipid accumulation through down-regulation of transcription factors such as PPARgamma and C/EBPalpha. 1,5-AF also induced dose-dependent phosphorylation of AMP-activated protein kinase (AMPK), a cellular energy sensor. However, the total AMPK protein content remained unchanged. Furthermore, 1,5-AF increased the levels of reactive oxygen species, an important upstream signal for AMPK activation in 3T3-L1 adipocytes. Our results show that 1,5-AF exerts anti-obesity action in vitro and suggest that 1,5-AF is potentially a novel preventative agent for obesity and other metabolic diseases.
A new pyrano[4,3-c][2]benzopyran-1,6-dione derivative antioxidant, named phelligridin K (1), was isolated from the methanol extract of the fruiting bodies of Fomitiporia ellipsoidea, together with the known compounds, (E)-4-(3,4-dihydroxyphenyl)but-3-en-2-one (2), hispidin (3), inoscavin C (4), and inonoblin B (5). Their structures were elucidated by means of extensive NMR and MS data analysis. Compounds 1, 4 and 5 exhibited significant scavenging activity against the ABTS radical cation.
A new 1,6-benzoxazocine-5-one alkaloid has been isolated as its butyl acetal derivative (1) along with peristrophine from the n-BuOH and EtOAc fractions of the crude MeOH extract of the aerial parts of Peristrophe lanceolaria growing in Thailand. The structures of these compounds were elucidated on the basis of their spectroscopic data. These compounds were isolated for the first time from P. lanceolaria. The EtOAc and n-BuOH fractions also possessed significant antioxidant activity with IC50 values of 57 and 50 μg/mL, respectively (DPPH method), whereas 1 had an IC50 value of 23 μg/mL.
Rabbit liver carboxylesterase (rCE) was evaluated as the catalyst for the enantioselective hydrolysis of (+/-)-3-endo-acetyloxy-1 ,8-cineole [(+/-)-4], which yields (1S,3S,4R)-(+)-3-acetyloxy-1,8-cineole [(+)-4] and (1R,3R,4S)-(-)-3-hydroxy-1,8-cineole [(-)-3]. Enantioselective asymmetrization of meso-3,5-diacetoxy-1,8-cineol (5) gives (1S,3S,4R,5R)-(-)-3-acetyloxy-5-hydroxy-1,8-cineole (6), with high enantioselectivity. rCE has been chosen to perform both experiments and molecular modeling simulations. Docking simulations combined with molecular dynamics calculations were used to study rCE-catalyzed enantioselective hydrolysis of cineol derivatives. Both compounds were found to bind with their acetyl groups stabilized by hydrogen bond interactions between their oxygen atoms and Ser221.
Comparison of observed (a) (1R,4R)-(+)-2 and (b) (1S,4S)-(-)2, and (c) calculated DFT B3LYP/DGDZVP VCD spectra of (1S,4S)-2. 
MMFF94 and DFT B3LYP/6-31G(d) relative energy, and DFT population for the most stable conformers of MTPA ester 6R and 6S. 
Enantiomerically pure (1S,4S)-(-)-3-oxo-1,8-cineole (-)-2 and (1R,4R)-(+)-3-oxo-1,8-cineole (+)-2 were prepared for the first time and their absolute configurations assigned by vibrational circular dichroism (VCD) measurements. Thus, treatment of cineole 1 with chromyl acetate gave rac-2 which after sodium borohydride reduction and acetylation provided racemic 3-endo-acetyloxy-1,8-cineole, rac-4. Enantioselective hydrolysis using porcine liver esterase (PLE) gave a mixture of 3-endo-hydroxy-1,8-cineole (-)-3 and 3-endo-acetyloxy-1,8-cineole (+)-4. After chromatographic separation, (-)-3 was oxidized to (+)-2, while (+)-4 was hydrolysed to (+)-3 and then oxidized to (-)-2. The absolute configuration of either ketone 2 was established by VCD spectroscopy in combination with density functional theory (DFT) calculations at the B3LYP/DGDZVP level of theory, from where it followed that the (+)-2 enantiomer corresponds to (1R,4R)-1,3,3-trimethyl-5-oxo-2-oxabicyclo[2.2.2]octane and the (-)-2 enantiomer to the (1S,4S) molecule which is also in agreement with the absolute configuration deduced by the Mosher method for the starting chiral alcohols. Some literature inconsistencies are clarified. In addition, the enantiomerically pure monoester (1S,3S,4R,5R)-(-)-3-acetyloxy-5-hydroxy-1,8-cineole 6 and the ketoester (1R,3S,4R)-(+)-3-acetyloxy-5-oxo-1,8-cineole 7 were prepared from meso-diacetate 5 by enantioselective asymmetrization also using PLE.
Chiral HPLC analyses on OJ-H column, eluted isocratically with n-heptaneEtOH, 95:5, at a flow rate of 1.0 mL/min. The elution was monitored at 210 nm. Graph A: Synthetic mixture of 3R and 3S; Graph B: falcarinol isolated from Patinaca sativa roots; Graph C: Synthetic mixture of 3R and 3S spiked with the isolated falcarinol from P. sativa roots. 
Falcarinol (9Z-heptadeca-1,9-diene-4,6-diyn-3-ol; (1) is a polyacetylene commonly found in several plant families. The absolute configuration of naturally occurring 1 is not clear and contradictory results have been reported in the literature. Determination of the absolute configuration of 1 from Pastinaca sativa L. was carried out. Isolation of 95% pure 1 was performed via successive fractionation and preparative-HPLC. A racemic mixture comprised of 3R-1 and 3S-1 was synthesized in order to confirm the absolute configuration of the isolated natural product using chiral HPLC. Based on a combination of chiral HPLC and specific rotation, 1 present in P. saliva was found to have a 3R absolute configuration (i.e. (3R, 9Z)-heptadeca-1,9-diene-4,6-diyn-3-ol).
Key NOESY correlations to determine the structure of compound 1.
Two novel cyclopeptides, xylarotides A (1), and B (2), and one novel nonenolide, xylarolide (3), along with two known compounds, coriloxin (4), and 2-hydroxy-3-methoxy-5-methyl-p-benzoquinone (5) were isolated from the fungal strain Xylaria sp. 101. This strain was isolated from the fruiting body of Xylaria sp. collected in Gaoligong Mountain, Yunnan Province. The chemical structures were elucidated by spectroscopic analyses, including 1D- and 2D-NMR spectroscopic experiments, and on the basis of HR-Q-TOF mass spectrometry. Antibacterial assays of 1 - 3 were carried out; no effects on the growth of the tested bacteria and yeast were observed.
Phytochemical investigation of the stems of Eurycoma longifolia Jack led to the isolation of two new canthin-6-one alkaloids, 4,9-dimethoxycanthin-6-one (1) and 10-hydroxy-11-methoxycanthin-6-one (2), and a new tirucallane-type triterpenoid, 23,24,25-trihydroxytirucall-7-en-3,6-dione (3), along with 37 known compounds. Among these, an oxasqualenoid (4) was isolated as a natural product for the first time. The structures of the isolates were elucidated by spectroscopic and mass spectrometric means. All the isolates were evaluated for their cytotoxic activity against a HT-1080 human fibrosarcoma cell line. Among them, 9,10-dimethoxycanthin-6-one (14, IC50 = 5.0 microM), 10-hydroxy-9-methoxycanthin-6-one (15, IC50 = 7.2 microM), dihydroniloticin (18, IC50 = 8.2 microM), and 14-deacetyleurylene (34, IC50 = 3.2 microM) displayed stronger activity than the positive control 5-FU (IC50 = 9.2 microM).
Chemical investigation of the leaves of Putranjiva roxburghii has resulted in the isolation of two new triterpenoids, putralone, a novel 10alpha-hydroxy-25-nor D:A friedo-olean-9(11)-en-3 one and 3beta-acetoxy-cycloart-24-en-23-one, along with a rare hopanoid, adian-5-en-3beta,29-diol. Other known triterpenoids isolated from this plant are 3beta-acetoxy-adian-5-ene, putrol, putrone, putranjivadione, roxburghonic acid, friedelin, friedlan-3alpha-ol, oleanolic acid and erythrodiol. Interestingly, putralone is the first example of a naturally occurring nor friedo-oleanane triterpenoid having a hydroxyl functionality at the C-10 position.
NOESY correlations of compound 10. 
13 C NMR spectroscopic data of compounds 3-8 in CDCl3. 
Scheme 1: Cyclization of cnicin (2). 
Scheme 2: Synthesis of compound 15. 
The acid-catalyzed cyclization of mono epoxides of cnicin acetonide (3) was investigated. Several 6,12-eudesmanolides were obtained, and their stereochemistry established by extensive spectroscopic analyses. Chemical correlations also led to the assignment of the absolute configuration of 1beta,10alpha-epoxy-salonitenolide (13), a previously isolated natural product. The cytotoxic activities of some compounds were determined against A549 and MCF-7 tumor cell lines. The esterified germacranolides 2-6 were selectively cytotoxic against the MCF-7 breast cancer cell line.
Anti-complementary activity-guided fractionation of the ethanolic extract of Prunella vulgaris spikes led to the isolation of a new, unusual delta11(12) triterpene, 3beta,13beta-dihydroxyolic-11-ene-28-oic acid (1), along with thirteen known triterpenes (2-14). The structure of the new compound was established by 1D and 2D NMR spectroscopic analysis. All the isolates were evaluated for their anti-complementary activity against the classical pathway (CP) and alternative pathway (AP). Eight triterpenes (1-8) showed anti-complementary activity against CP and AP, with CH50 and AP50 values of 0.15-0.37 mg/mL, and 0.29-0.53 mg/mL, respectively. Mechanism study using complement-depleted sera showed that oleanolic acid (2) acted selectively on C1q, C5, and C9 components, while 2alpha-hydroxy oleanolic acid (3) interacted with C1q, C3, C5 and C9, and 2alpha,3alpha-dihydroxyolic-12-ene-28-oic acid (4) blocked C1q, C2, C3, C5 and C9.
Three new natural furanones, pulvinulin A (1), graminin C (2), and cis-gregatin B (3), together with the known fungal metabolites, graminin B (4) and 10-norparvulenone (5), were isolated from Pulvinula sp. 11120, an endophytic fungal strain occurring in healthy foliage of Cupressus arizonica (Arizona cypress). The structures of 1 and 2 were elucidated by the analysis of their spectroscopic data and chemical interconversions, and that of 3 was determined by comparison with data for synthetic cis-gregatin B. Comparison of spectroscopic data of 4 and 5 with those reported identified them as graminin B and 10-norparvulenone, respectively. Metabolites 1-4 exhibited antibacterial activity against E. coli.
Two new angucyclines, saccharothrixmicines A (2) and B (3), together with three known diketopiperazines 4-6 were isolated from the actinomycete Saccharothrix espanaensis An 113 associated with the marine mollusk Anadara broughtoni. Their structures were determined by HRESI-MS and 1D and 2D NMR. Compounds 2 and 3 differ in aglycone and glycosidic bond type. 2 is an alpha-L-6-deoxyaltrose-phenylglycoside of a benz[a]anthraquinone aglycone, while 3 is an O-glycoside of the same sugar linked to C-7 of the known angucyclinone (1). A saccharothrixmicine-containing fraction exhibited activity towards Candida albicans and Xanthomonas sp. pv. badrii whereas the diketopiperazines showed antibiotic activities against Vibrio alginolyticus and Vibrio parahaemolyticus.
Limonoids and triterpenes are the largest groups of secondary metabolites and have notable biological activities. Meliaceae and Rutaceae are known for their high diversity of metabolites, including limonoids, and are distinguished from other families due to the frequent occurrence of such compounds. The increased interest in crop protection associated with the diverse bioactivity of these compounds has made these families attractive in the search for new allelopathic compounds. In the study reported here we evaluated the bioactivity profiles of four triterpenes (1-4) and six limonoids (5-10) from Meliaceae and Rutaceae. The compounds were assessed in a wheat coleoptile bioassay and those that had the highest activities were tested on the standard target species Lepidinum sativum (cress), Lactuca sativa (lettuce), Lycopersicon esculentum (tomato) and Allium cepa (onion). Limonoids showed phytotoxic activity and 5α,6β,8α, 12α- tetrahydro-28-norisotoonafolin (10) and gedunin (5) were the most active, with bioactivity levels similar to, and in some cases better than, those of the commercial herbicide Logran. The results indicate that these products could also be allelochemicals involved in the ecological interactions of these plant species.
The composition of 16 samples of leaf oil from Corsican Juniperus thurifera was investigated by integrated techniques, GC, GC-MS and 13C NMR. K-means partitioning and PCA analysis of the data allowed the definition of a main group (14 samples) dominated by limonene (mean = 52.2%, SD = 6.4) and alpha-pinene (mean = 7.2%, SD = 3.8). Limonene and beta-elemol (up to 19.7%) were identified as the major components of two atypic samples.
Both enantiomers of 12-methyl-13-tridecanolide{(R)-(+)-1, (S)-(-)-1} and 14-methyl-15-pentadecanolide (muscolide) {(R)-(+)-2, (S)-(-)-2} were synthesized from either (S)-(+)- or (R)-(-)-3-bromo-2-methyl-1-propanol 8 as a chiral building block.
Epimeric (20R,23R)- and (20R,23S)-23-hydroxycholestane steroids were synthesized. Their structures were elucidated by extensive 1H and 13C NMR spectroscopy and application of the Mosher's method. All proton and carbon signals of the side chains were assigned. Based on these assignments spectral data allow the determination of the C-23 stereochemistry of (20R)-23-hydroxycholestane side chains of the new natural steroids by comparison with spectra of the obtained model compounds. As a result, the C-23 configuration of two steroid compounds from the starfishes Lethasterias nanimensis chelifera and Lethasterias fusca was established.
This study was designed for simultaneous determination of 13 chemical marker compounds, namely, protocatechuic acid, chlorogenic acid, caffeic acid, liquiritin, hesperidin, apigetrin, rosmarinic acid, oxypeucedanin hydrate, byakangelicin, apigenin, glycyrrhizin, nobiletin, and 6-gingerol in Gwakhyangjeonggi-san (GJS: Huoxiang-zhengqi-san in Chinese). A quantitative analytical method was developed based on HPLC-PDA with validation in terms of precision, accuracy, and repeatability, and successfully employed for quality evaluation of GJS samples with the help of chemometric techniques such as principal component analysis (PCA) and hierarchical clustering analysis (HCA). The correlation coefficient for the linear regression was > 0.9994. The intra-day and inter-day precision was < 3.0% of the relative standard deviation (RSD) value, and the recovery was in the range 92.5-107.0%, with RSD values < 4.0%, and the repeatability was < 3.0% of RSD. Variations in the quantity were observed in GJS products from different origins, which were classified by PCA and HCA. The quantitative and chemometric analyses indicate the necessity for consistency in GJS production for the purpose of quality control.
Partial HSQC spectrum of resin sample V1. P = pinoresinol, L = lariciresinol, La= lariciresinol acetate. 
Identification and quantitative determination of individual components of resin collected on the trunk of 28 Cedrus atlantica trees, grown in Corsica, has been carried out using 13C NMR spectroscopy. Eight resin acids bearing either the pimarane or abietane skeleton, two monoterpene hydrocarbons and four oxygenated neutral diterpenes have been identified, as well as three lignans, scarcely found in resins. Three groups could be distinguished within the 28 resin samples. The nine samples of Group I had their composition dominated by diterpene acids (33.7-45.8%), with abietic acid (6.2-18.7%) and isopimaric acid (5.1-12.6%) being the major components. The four samples of Group II contained resin acids (main components) and lignans in moderate amounts (up to 10.3%). Conversely, lignans (38.8-63.8%) were by far the major components of the 15 samples of Group III. Depending on the sample, the major component was pinoresinol (18.1-38.9%), lariciresinol (17.2-33.7%) or lariciresinol 9'-acetate (16.9-29.1%). Finally, due to the high biological interest in lignans, a rapid procedure, based on 1H NMR spectroscopy, was developed for quantification of lignans in resins of C. atlantica.
Ozonolysis of (+)-manool (4) in different solvents. 
14,15,17-Trinorlabdan-8,13-dione 6 was efficiently synthesized via ozonolysis of(+)-manool (4) followed by treatment with aqueous NaOH in the presence of tetra-n-butylammonium bromide as catalyst. This protocol has the advantages of high yield, mild conditions and simple procedure. Utilizing this strategy, the first enantiospecific synthesis of 13,14-dihydroxy-8,11,13-podocarpatrien-7-one (1), a constituent of Taiwania cryptomerioides and Celastrus paniculatus, was achieved starting from (+)-manool (4) after a four-step sequence in 24% overall yield.
Radiotherapy is an important form of treatment for a wide range of cancers, but it can damage DNA and cause adverse effects. We investigated if the diplacone analogs of P. tomentosa were radio-protective in a human lymphoblastoid cell line (AHH-1). Four geranylated flavonoids, diplacone, 3'-O-methyl-5'-hydroxydiplacone, 3'-O-methyl-5'-O-methyldiplacone and 3'-O-methyldiplacol, were tested for their antioxidant and radio-protective effects. Diplacone analogs effectively scavenged free radicals and inhibited radiation-induced DNA strand breaks in vitro. They significantly decreased levels of reactive oxygen species and cellular DNA damage in 2 Gy-irradiated AHH-1 cells. Glutathione levels and superoxide dismutase activity in irradiated AHH-1 cells increased significantly after treatment with these analogs. The enhanced biological anti-oxidant activity and radioprotective activity of diplacone analogs maintained the survival of irradiated AHH-1 cells in a clonogenic assay. These data suggest that diplacone analogs may protect healthy tissue surrounding tumor cells during radiotherapy to ensure better control of radiotherapy and allow higher doses of radiotherapy to be employed.
The essential oil (EO) of M. aquatica L. growing wild in Corsica was isolated by dry vapor distillation and submitted to combined analysis by column chromatography over silica gel, GC(RI), GC-MS and 13C NMR spectroscopy. The composition was dominated byoxygenated monoterpenes and characterized by the occurrence of menthofuran (50.7%) as the major component. In parallel, seven laboratory-distilled oil samples isolated from individual plants collected in Corsica were analyzed by GC(RI) and 13C NMR spectroscopy. Onlyquantitative differences were observed between the samples. Beside the usual terpenes, various p-menthane lactones (mintlactone, isomintlactone, hydroxymintlactone, menthofurolactone and epimenthofurolactone) have been identified in all the oil samples.
Different techniques have been utilized to determine the composition of Artemisia absinthum (wormwood) essential oil. The oil was fractionated on a silica gel column and each fraction analyzed by GC(RI), GC-MS and ¹³ C NMR. This allowed the identification, for the first time in A. absinthium, of two diterpenes, 9-geranyl-p-cymene and 9-geranyl-α-terpinene, and two homoditerpenes, 9-(15,16-dihydro-15-methylene)-geranyl-p-cymene and 9-(15,16-dihydro-15-methylene)-geranyl-α-terpinene. Chemical variability of A. absinthium essential oils from plants collected in the surroundings of Vilnius city over several years (1999-2007) was also shown. Chemical composition was determined by GC and GC-MS. Thujones ( cis+trans, 10.2-36.3%) and trans-sabinyl acetate (9.8-39.2%) were the two predominant constituents of almost all the investigated oils (13 out of 15 samples). The third major compound was myrcene (5.1-9.2%, in four samples), β-pinene (5.4-10.4%, in 5), linalool (4.7% in one), trans-sabinol (6.4%, in one) and 1,8-cineole (5.2-7.1%, in two). In one oil, the prevailing components were thujones ( cis+trans, 11.2%), trans-sabinene hydrate (11.0%) and trans-sabinyl acetate (8.8%), while another sample was characterized by a large quantity of trans-sabinyl acetate (55.2%) and the absence of thujones.
Two samples (leaves and twigs) of Abies alba Miller from Corsica were extracted using supercritical CO2 and their chemical compositions were compared with those of the essential oils obtained from the same batch of plant material. In total 45 components were identified using combined analysis by GC (RI), GC-MS and 13C NMR. It was observed that the contents of monoterpenes (mainly represented by limonene, alpha-pinene and camphene) were significantly lower in the supercritical fluid extract (SFE) than in the essential oil (EO). Conversely, the proportions of sesquiterpenes were much higher in CO2 extracts than in essential oils (around 30% vs 4%). Cis-abienol, a diterpene alcohol, was identified only in SFE, and the proportions of this constituent (7.5% and 17.3%) were determined using quantitative 13C NMR since it was under estimated using the standard conditions of GC.
Morphology of GL-15 cells in control conditions (A) and after 24 h exposure to 250 µg/mL AEP-1 after immunocytochemistry for the cytoskeleton proteins GFAP (red), and α-tubulin (green), and nuclear chromatin staining with Hoechst-33258 (blue).  
Nuclear morphology of GL-15 cells in (A) control conditions or after 72 h exposure to 250 µg/mL of (B) AEP-1 after chromatin staining with Hoechst-33258 (blue).  
Electrophoresis under alkaline conditions in low melting point agarose gel on the GL-15 cells in control condition and after 72 h exposure to AEP-1.  
Cytokine concentrations in culture medium of glial cells after 72 h experiment. Paired T-tests were performed to measure the significance of the difference of the mean between negative control (NC) and treated cultures (250 µg/mL). Only the cytokines with a percentage of coefficient of variation higher than 20% are shown.  
Plant secondary metabolites, such as, specifically, alkaloids and terpenes, may present psychoactive properties that modify the function of the central nervous system (CNS) and induce neurotoxicity. Neurotoxicity involves the response of glial cells, mainly astrocytes, which play a fundamental role in the control of homeostasis of the CNS. Some Erythroxylum species are indigenous to the state of Bahia in Brazil. This study investigated the cytotoxic activity of the diterpene AEP-1, extracted from the fruit of E. passerinum in a GL-15 cell line, astrocytic, glial cells model. The effects on cell viability, analyzed by the MTT assay, demonstrated a dose-dependent cytotoxic effect, with maximum effect at 500 μg/mL of AEP-1, and with a reduction of about 40 and 47% on cellular viability after 24 h and 72 h treatment, respectively. Evidence for induction of apoptosis by AEP-1 was first obtained when GL-15 glial cells were incubated with 250 μg/mL AEP-1 causing reniform and/or pyknotic nuclei and apoptotic bodies revealed by chromatin staining with Hoechst 33258. Increase in DNA fragmentation was also observed by comet assays in cells incubated with 500 μg/mL of AEP-1. Moreover, cells exposed to a sub toxic dose of AEP-1 (250 μg/mL) showed significant changes in morphology--contraction of the cytoplasm and expansion of cellular projections--signifying the presence of astrocytic cytoskeletal protein and glial fibrillary acidic protein (GFAP). These findings indicated astrocytic cells as the target for terpene AEP-1 and suggest the involvement of glial cells with psychoactive symptoms observed in humans and animals after consumption of fruits of plants of the genus Erythroxylum.
Recent published sequencing of fungal genomes has revealed that these microorganisms have a surprisingly large number of secondary metabolite pathways that can serve as potential sources for new and useful natural products. Most of the secondary metabolites and their biosynthesis pathways are currently unknown, possibly because they are produced in very small amounts and are thus difficult to detect or are produced only under specific conditions. Elucidating these fungal metabolites will require new molecular genetic tools, better understanding of the regulation of secondary metabolism, and state of the art analytical methods. This review describes recent strategies to mine the cryptic natural products and their biosynthetic pathways in fungi.
Potencies of DCTA/DKA hybrid inhibitors against recombinant IN K156A-, K159A-, and reference IN-mediated strand transfer. 
Increasingly non-conservative substitution of K156 and K159 confers attenuated 3'-end processing and strand transfer specific activities. 3'-End processing (black bars) and strand transfer (white bars) enzyme specific activities for recombinant reference IN, from HIV NL4-3 , and each mutant protein were determined. Specific activities were calculated via linear regression analysis of a plot of pmol IN versus fmol products generated per hour. Bars indicate the mean of triplicate assays; error bars indicate 1 SD. 
Representative strand transfer IC 50 assays-IN K159A confers high resistance to L-731,988. 100 nM Recombinant reference IN (A.), IN K159R (B.), IN K159A (C.), or IN K159D (D.) was incubated with 5 nM radiolabeled strand transfer oligonucleotide duplex substrate, U5V1P/V2, in increasing concentrations of L-731,988 at 37°C for 1 hour, in triplicate. Reactions were terminated by addition of EDTA to 50 mM, resolved via denaturing PAGE, then visualized and quantified via phosphorimager analysis. IC 50 values were calculated using CalcuSyn software for windows.-IN is substrate alone. +IN is reaction with IN and no inhibitor. (-) contains 25 μM L-tartaric acid, an inactive compound. (+) contains 25 μM L-CA, a concentration that completely inhibits INmediated strand transfer catalysis. Numbers above triplicates indicate concentrations of L-731,988 in μM. Solid arrows indicate unprocessed substrate and dashed vertical lines indicate strand transfer products. 
Human immunodeficiency virus (HIV) type I integrase (IN) active site, and viral DNA-binding residues K156 and K159 are predicted to interact both with strand transfer-selective IN inhibitors (STI), e.g. L-731,988, Elvitegravir (EVG), and the FDA-approved IN inhibitor, Raltegravir (RGV), and strand transfer non-selective inhibitors, e.g. dicaffeoyltartaric acids (DCTAs), e.g. L-chicoric acid (L-CA). To test posited roles for these two lysine residues in inhibitor action we assayed the potency of L-CA and several STI against a panel of K156 and K159 mutants. Mutagenesis of K156 conferred resistance to L-CA and mutagenesis of either K156 or K159 conferred resistance to STI indicating that the cationic charge at these two viral DNA-binding residues is important for inhibitor potency. IN K156N, a reported polymorphism associated with resistance to RGV, conferred resistance to L-CA and STI as well. To investigate the apparent preference L-CA exhibits for interactions with K156, we assayed the potency of several hybrid inhibitors containing combinations of DCTA and STI pharmacophores against recombinant IN K156A or K159A. Although K156A conferred resistance to diketo acid-branched bis-catechol hybrid inhibitors, neither K156A nor K159A conferred resistance to their monocatechol counterparts, suggesting that bis-catechol moieties direct DCTAs toward K156. In contrast, STI were more promiscuous in their interaction with K156 and K159. Taken together, the results of this study indicate that DCTAs interact with IN in a manner different than that of STI and suggest that DCTAs are an attractive candidate chemotype for development into drugs potent against STI-resistant IN.
A new environmentally benign and highly efficient catalytic system [pi-C5H5NC16H33]3[PO4(WO3)4]/H2O2/CH3COOC2H5 for the epoxidation of soybean oil displayed excellent activity and high recovery. The change of the catalyst during the reaction was investigated by elemental analysis, FT-IR and 31P NMR.
Four new triterpene glycosides cucumariosides H5 (1), H6 (2), H7 (3) and H8 (4) along with the known cucumarioside H (5) have been isolated from the Far Eastern sea cucumber Eupentacta fraudatrix. The structures of glycosides 1-4 were elucidated on the basis of spectral data (2D NMR and MS). Glycosides 1-4 belong to the group o f cucumariosides H having branched rare pentasaccharidecarbohydrate moieties with one sulfate group and 3-O-methyl-D-xylose as a terminal monosaccharide unit. Glycosides 1-3 and 5 differ from each other in structures of side chains of the aglycones, while cucumarioside H8 (4) has a novel aglycone with unprecedented 16(22)-epoxy-group, never found in the sea cucumbers glycosides. Glycosides 1-3, and 5 were cytotoxic against mouse lymphocytes and hemolytic against mouse erythrocytes. Glycoside 2 was less active in comparison with others.
Szentiamide (1) a new cyclic hexadepsipeptide was isolated from the culture broth of the entomopathogenic bacterium Xenorhabdus szentirmaii DSM 16338T. The structure was elucidated by analysis of one- and two-dimensional NMR spectra and high resolution mass spectrometry. The amino acids were determined to be D-leucine, L-threonine, D-phenylalanine, D-valine, L-tyrosine and L-tryptophane after hydrolysis and derivatization with D-FDVA [Nalpha-(2,4-dinitro-5-fluorophenyl)-D-valinamide].
Biotransformation of 16alpha,17-epoxy-ent-kaurane-19-oic acid (1) by Beauveria sulfurescens ATCC 7159-F led to the production of a new ent-kaurane diterpenoid, 7beta,17-dihydroxy-ent-kaur-15-en-19-oic acid (7), and four other ent-kauranes (8 - 11), all of which were identified as their methyl esters. Compounds 9 and 10 were found to be new stereoisomers. Structures of these were established by the extensive usage of their spectroscopic characteristics.
Seven new minor triterpene glycosides, cucumariosides A2 (1), A7 (2), A9 (3), A10 (4), A11 (5), A13 (6) and A14 (7) have been isolated from the Far Eastern sea cucumber Eupentacta fraudatrix. Structures of the glycosides were elucidated by 2D NMR spectroscopy and MS. Glycosides 1-7 belong to the group of cucumariosides A, having linear tetrasaccharide carbohydrate moieties without any sulfate group and possessing 3-O-methyl-D-xylose as a terminal monosaccharide unit. Glycosides 1, 2, 5-7 differ from each other in side chain structures in aglycone moieties, while cucumarioside A10 (4) has a 23,24,25,26,27-pentanorlanostane aglycone with 18(16)-lactone. Cucumarioside A9 (3), having an uncommon 18-hydroxy group, is the second representative of the unique metabolically active glycosides that are regarded as intermediates of glycoside biosynthesis in sea cucumbers. Cytotoxic activities of glycosides 1-7 and cucumarioside A8 (8) against mouse spleen lymphocytes and the cells of the ascite form of mouse Ehrlich carcinoma, along with hemolytic activity against mouse erythrocytes and antifungal activity were studied. Cucumariosides A2 (1), A8 (8) and A13 (6) demonstrated high hemolytic activities. Glycosides 1, 4 and 6 showed moderate cytotoxic activity. Only cucumarioside A8 (8), having an 18-oxymethylene group and a 24(25)-double bond, was very active in all the tests.
Three new 18-oxygenated ent-kaurane diterpenoids, isoleuconins A-C (1-3) and ten known diterpenoids were isolated from the aerial parts of Isodon leucophyllus. The structures were elucidated by 1D and 2D NMR spectroscopic analysis. All of the compounds were evaluated for their cytotoxicity. Rabdokunmin A (13) showed significant cytotoxicity against HT-29 cells, with an IC50 value of 6.2 microM.
Thirty three C19-diterpenoid alkaloids, twenty-two prepared from known C19-diterpenoid alkaloids and eleven isolated from Aconitum and Delphinium spp. were evaluated for their cardiac activity in the isolated bullfrog heart assay. Among them, eleven compounds exhibited cardiac activity, with average rate of amplitude increase in the range of 16-118%. Compound 7, mesaconine (17), hypaconine (25), and beiwutinine (26) exhibited strong cardiac activities relative to the reference drug. The structure-activity relationship data acquired indicated that an alpha-hydroxyl group at C-15, a hydroxyl group at C-8, an alpha-methoxyl or hydroxyl group at C-1, and a secondary amine or N-methyl group in ring A are important structure features necessary for the cardiac activities of the aconitine-type C19-diterpenoid alkaloids without any ester groups. In addition, an alpha-hydroxyl group at C-3 is also helpful for the cardiac activity of these alkaloids.
Treatment of a 7,17-seco-type C19-diterpenoid alkaloid (3), prepared from deltaline (8), with triethylamine in either DMF or TEG (triethylene glycol) at 120 degrees C provided two interesting compounds 6 and 7. The structures of compounds 3, 6, and 7 were established based on extensive interpretations of their 1D and 2D NMR data. Compound 6, a lycoctonine-type C19-diterpenoid alkaloid, can be transformed from alkaloid 3 via Grob fragmentation, Prins reaction, and intramolecular disproportionation. The mechanism of the formation of compound 6 was confirmed by deuteration experiments. Product 7 was formed through a pinacol-like rearrangement of alkaloid 3.
19-OH-(-)-eburnamonine 1 is a new indole alkaloid isolated from Bonafusia macrocalyx. A natural derivative from (-)-eburnamonine for which absolute configuration and conformation has been determined by making use of residual dipolar couplings enhanced NMR, circular dichroism spectra and high-level computations.
A novel access to 7,17-seco Cl9-diterpenoid alkaloids via vacuum pyrolysis of N-deethyl-8-acetyl derivatives is described.
Several species belonging to the Hyptis (Lamiaceae) genus represent an important source of bioactive constituents, which are reputed for their wide range of antimicrobial, anticancer and insecticidal activities. The volatile oils obtained from various parts of the Hyptis plants were found to be primarily composed of mono- and sesquiterpenes. Significant differences were observed in the percentage compositions of the major components, which allowed for differentiation among the species. Based on the dominant constituents, phylogenetic relationships were found to be common among some species: 1,8-cineole (H. fruticosa, H. goyazensis, H. martiusii and H. suaveolens); beta-caryophyllene (H. marrubioides, H. pectinata, H. spicigera and H. suaveolens); eugenol (H. recurvata and H. suaveolens); gamma-cadinene (H. glomerata and H. ovalifolia); p-cymene (H. mutabilis and H. pectinata); alpha-pinene (H. crenata and H. emoryi). The monoterpenes, alpha-pinene and p-cymene were detected at various concentrations in all the Hyptis oils investigated. This paper reviews the essential oil compositions of eighteen Hyptis plant species studied in various parts of the world covering the publications of 100 years (1909 to 2009) after the first article appeared in the literature.
Eichhornia crassipes (Mart.) Solms (Waterhyacinth), an aquatic perennial herb present throughout the world, has a myriad of metabolites. Phenalenone compounds and sterols have been isolated from this plant. Extracts, as well as pure compounds isolated from this plant, have been demonstrated to possess pharmacological activities. An account of the phytochemistry, pharmacological activities and several applications ofwaterhyacinth are included in this review.
A methanol extract of the leaves of Blumea balsamifera (L.) DC. (Asteraceae) afforded a new guaian-type sesquiterpene, epiblumeaene K (1), together with four known guaian-type sesquiterpenes (2-5), three known sesquiterpenes (6-8), and nine known flavonoids (9-17) by a combination of chromatography and preparative TLC techniques. Their structures were elucidated by extensive spectroscopic methods and comparison with the literature data. Among the isolated compounds, a known sesquiterpene, beta-caryophyllene 8R,9R-oxide (6), exhibited a significant PTP1B inhibitory activity in a dose-dependent manner, with an IC50 value of 25.8 microM (5.62 microg/mL).
Observed vibrational spectra of 3α,6β-di(1-methyl-1H-pyrrol-2- ylcarbonyloxy)tropane (2). VCD spectra of the (-)-(3R,6R) isomer (top) and the (+)-(3S,6S) isomer (center), and IR spectrum (identical for both isomers) (bottom).  
Observed VCD spectra of (-)-(2´S)-scopolamine (top) and (-)-(2´S)- hyoscyamine (bottom). tropane bicycle. As observed in the VCD spectra, neither shows the characteristic patters of tropanediol derivatives, further confirming the validity of the empirical rule used herein.  
Chiral resolution of (+/-)-3alpha,6beta-dicinnamoyloxytropane (1) and (+/-)-3alpha,6beta-di(1-methyl-1H-pyrrol-2-ylcarbonyloxy)tropane (2), prepared by esterification of (+/-)-3alpha,6beta-tropanediol (3), was achieved using an amylose-derived HPLC stationary phase and normal phase conditions. The corresponding vibrational circular dichroism (VCD) spectra provided the absolute configuration of the enantiomers as (-)-(3R,6R)-1, (+)-(3S,6S)-1, (-)-(3R,6R)-2 and (+)-(3S,6S)-2. In each case, characteristic VCD bands for the absolute configuration determination of the 3alpha,6beta-tropandiol esters were observed. While the absolute configuration of natural 1, previously isolated from Erythroxylum hypericifolium, could not be established due to the lack of literature optical rotation values, that of catuabine E, previously isolated from E. vacciniifolium, is now assigned as (-)-(3R,6R)-2 by comparison with the optical rotation values of the prepared samples and the reported rotation of the natural product.
The first synthesis of isochromene fused carbazols, (4aS, 13bR)-2,5,5-trimethyl-3,4,4a,5,8,13b-hexahydroisochromeno[3,4-b]carbazole (2) and its epi-isomer 3 by condensation of citral and 2-hydroxycarbazole using Ti(OEt)4 and MeAlC12 as catalysts is described.
Fungicidal activities of the synthesized compounds (% control).
A series of new strobilurin derivatives with a substituted pyrazole side chain (5 and 6) were designed and synthesized. The new derivatives were synthesized from substituted phenones as starting material via esterification, cyclization and condensation reactions. All compounds were identified by 1H NMR and IR spectral and elemental analyses. Preliminary biological evaluation showed that some of the compounds had good fungicidal activity against cucumber downy mildew (CDM) and wheat powdery mildew (WPM) at 6.25 and 1.56 mg L(-1), respectively. Some of the compounds showed insecticidal activity against armyworm (AW), green peach aphid (GPA) and culex mosquitoes at 600 mg L(-1).
Top-cited authors
William N Setzer
  • University of Alabama in Huntsville
Michael Wink
  • Universität Heidelberg
Alvaro Viljoen
  • Tshwane University of Technology
Wan Mohd Nuzul Hakimi Wan Salleh
  • Sultan Idris Education University (UPSI)
Debra Moriarity
  • University of Alabama in Huntsville