Sesquiterpene Polygodial from Drimys brasiliensis (Winteraceae) Down-Regulates Implant-Induced Inflammation and Fibrogenesis in Mice.

Drimys brasiliensis (Winteraceae) has been investigated in traditional medicine for its anti-inflammatory properties to treat gastric ulcers and allergic and respiratory system diseases as well as for cancer treatment. In this work, we investigate the ability of the sesquiterpene polygodial, isolated from D. brasiliensis stem barks, to modulate the chronic inflammatory response induced by polyester-polyurethane sponge implants in C57BL/6J mice. Daily treatment with polygodial inhibited the macrophage content in the implants as determined by the activity of the N-acetyl-ß-d-glucosaminidase enzyme as well as decreased the levels of CXCL1/KC and CCL2/JE/MCP-1 pro-inflammatory chemokines and the presence of mast cells along the formed fibrovascular tissue. Similarly, the deposition of a new extracellular matrix (total collagen and type I and III collagen fibers) as well as the production of the TGF-ß1 cytokine were attenuated in implants treated with polygodial, showing for the first time its antifibrogenic capacity. The hemoglobin content, the number of newly formed vessels, and the levels of VEGF cytokine, which were used as parameters for the assessment of the neovascularization of the implants, did not change after treatment with polygodial. The anti-inflammatory and antifibrogenic effects of polygodial over the components of the granulation tissue induced by the sponge implant indicate a therapeutic potential for the treatment of inflammatory diseases associated with the development of fibrovascular tissue.

Anti-diabetic and lipid-lowering effects of drimane sesquiterpenoids isolated from Zygogynum pancheri.

Recently, it has been shown that drimane-type sesquiterpenoids isolated from Zygogynum pancheri, a species native to New Caledonia, possessed significant α-amylase inhibitory activities. To further explore their antidiabetic potential, we investigated the effect of 1ß-O-(E-cinnamoyl)-6α-hydroxy-9epi-polygodial (D) and 1ß-E-O-p-methoxycinnamoyl-bemadienolide (L), two of the most active compounds of the series, on diabetic model rats. Compounds D and L (2 mg kg/day) were daily and orally administrated for 30 days to streptozotocin (STZ) (150 mg/kg) induced male diabetic Wistar rats. Animals were allocated into five groups of six rats. Comparatively to diabetic rats, treatments with D and L compounds were able to significantly (P < 0.05) decrease Fasting Blood Glucose (FBG) (70.15%, 71.02%), serum total cholesterol (46.27% and 39.38%), triglycerides (56.60% and 58.15%), creatinine (37.31% and 36.49%) and uric acid levels (67.76% and 69.68%), respectively. Compounds D and L also restored the altered plasma enzyme (aspartate aminotransferase, AST (47.83% and 43.20%), alanine aminotransferase, ALT (49.76% and 48.35%, alkaline phosphatase, ALP (72.78% and 73.21%)) and lactate dehydrogenase, LDH (47.95% and 53.93%) levels to near normal, respectively. Administration of Glymepiride, significantly (p < 0.05) reduced FBG (73.94%) in STZ induced diabetic rats. Additionally, the compounds D and L exhibited inhibitory effects in vivo on lipase activity of diabetic rats (54.83% and 52.25%), respectively. The outcomes of this study suggested that these two drimanes could be considered as efficient hypoglycemic, hypolipidemic and antiobesity agents for diabetes management and its complications.

Gold nanoparticle-assisted enhancement in bioactive properties of Australian native plant extracts, Tasmannia lanceolata and Backhousia citriodora.

Green nanotechnology plays a significant role in developing effective treatment strategies for numerous diseases. The biological synthesis of metal nanoparticles (M-NPs) possesses suitable alternatives than chemical techniques. Using plant extract to synthesis M-NPs is an eco-friendly, non-toxic, and cost-effective that are suitable for biological applications and efforts are directed to explore the efficacy of these materials in cancer management. In this study, gold nanoparticles (Au-NPs) were synthesised by following a one-step green synthesis, a reaction between HAuCl4 and biological molecules present in Tasmannia lanceolata leaf extract as a sole agent for both reduction and stabilisation. The characterisation techniques confirmed the successful synthesis of Au-NPs. TEM photograph revealed spherical shape nanoparticles with an average size of 7.10 ± 0.66 nm. The in-vitro cytotoxicity of Au-NPs was performed by analysing the percentage inhibition of cell viability using Resazurin assay on human liver cancer (HepG2), melanoma cancer (MM418 C1) and breast cancer (MCF-7) cell lines and compared with Au-NPs synthesised by using Backhousia citriodora leaf extract. The results showed that biosynthesised Au-NPs displayed greater inhibitory activity towards MCF-7 cancer cells proliferation compared to HepG2 and MM418 cancer cells. In addition, synthesised Au-NPs@ Tasmannia lanceolata leaf extract indicated higher inhibitory activity towards cancer cells compared to Au-NPs@ Backhousia citriodora leaf extract.

Distinct Drimane Chemotypes in Tasmanian Mountain Pepper (Tasmannia lanceolata): Differences in the Profiles of Pungent Leaf Phytochemicals Associated with Altitudinal Cline.

This study assesses whether the distinct altitudinal cline in leaf morphology (decreased leaf width and length with increased altitude) in Tasmanian mountain pepper (Tasmannia lanceolata) is associated with changes in the leaf chemistry of wild populations from different ecological landscapes and altitudes. The presence of distinct pungent drimane sesquiterpenoid chemotypes was identified: subalpine woodland and wet sclerophyll forest chemotypes. Isolation studies and analysis of extracts revealed that wet sclerophyll forest T. lanceolata populations featured polygodial as the principal terpenoid, with profiles similar to the commercial cultivars sampled. In contrast, the subalpine woodland populations contained the drimane sesquiterpenoids 1ß-acetoxy-9-deoxyisomuzigadial and 3ß-acetoxydrimenin and the conspicuous absence of the pungent principle polygodial.

Inhibition of the growth of human dermatophytic pathogens by selected australian and asian plants traditionally used to treat fungal infections.

OBJECTIVE: Syzygium australe (H.L. Wnddl. ex. Link) B. Hyland, Syzygium luehmannii (F. Muell.) L.A.S. Johnson, Syzygium jambos L. (Alston), Terminalia ferdinandiana Exell. and Tasmannia lanceolata (Poir.) A.C.Sm. are used in traditional Australian Aboriginal and Asian healing systems to treat a variety of pathogenic diseases including fungal skin infections, yet they are yet to be examined for the ability to inhibit the growth of human dermatophytes. MATERIALS AND METHODS: The fungal growth inhibitory activity of extracts produced from selected Australian and Asian plants was assessed against a panel of human dermatophytes by standard disc diffusion and liquid dilution MIC methods. The toxicity of the extracts was evaluated by Artemia lethality and MTS HDF cell viability assays. The phytochemistry of the most promising extracts were examined by GC-MS headspace analysis and some interesting compounds were highlighted. RESULTS: The aqueous and methanolic extracts of all plant species were good antifungal agents, inhibiting the growth of all of the dematophytes tested. The methanolic S. australe (SA) and S. luehmannii (SL) extracts were particularly potent fungal growth inhibitors. MIC values of 39 and 53µg/mL were recorded for the methanolic SL fruit extract against T. mentagrophytes and T. rubrum respectively. Similar MICs were also noted for the methanolic SL leaf extract (88 and 106µg/mL respectively). The methanolic SL leaf extract was a particularly good fungal growth inhibitor, with MIC values≤100µg/mL against the reference C. albicans strain (96µg/mL), E. floccosum (53µg/mL), and T. mentagrophytes (88µg/mL). This extract also produced MICs≤200µg/mL against all other fungal species/strains tested. Similarly good activity was seen for the methanolic S. australe leaf and fruit extracts, as well as the S. lehmannii fruit and S. jambos leaf extracts, with MIC values 100-500µg/mL. Interestingly, these extracts had low toxicity and high therapeutic indices, indicating their suitability for clinical use. GC-MS headspace analysis highlighted several monoterpenoids and sesquiterpenoids in the methanolic SA and SL extracts. T. ferdinandiana and T. lanceolata extracts also had promising antifungal activity, albeit with substantially higher MICs. CONCLUSION: Whilst multiple extracts inhibited fungal growth, the methanolic S. australe and S. luehmannii leaf extracts and the S. luehmannii fruit extracts showed particularly potent activity against each of these dermatophytes, indicating that they are promising leads for the development of anti-dermatophytic therapeutics.

Structure-Pungency Relationships and TRP Channel Activation of Drimane Sesquiterpenes in Tasmanian Pepper (Tasmannia lanceolata).

Sensory-guided fractionation of extracts of Tasmanian pepper berries revealed 20 drimane sesquiterpens, among which polygodial, warburganal, and 1ß-acetoxy-9-deoxy-isomuzigadial exhibited the lowest pungency threshold concentrations on the tongue surface (0.6-2.8 nmol/cm2) and elicited a dose-dependent calcium influx into mTRPA1 expressing CHO cells with the lowest EC50 values (4.5 ± 1.0 to 16.7 ± 7.5 µmol/L) and a good correlation to oral pungency thresholds (R2 = 0.986, linear regression). Calcium imaging assays demonstrated these chemosensates to induce a calcium influx into cultured trigeminal neurons prepared from wildtype (TRPA1+/+) mice, whereas no calcium influx was observed in neurons from TRPA1 knockout mice (TRPA1-/-), thus confirming the α,ß-unsaturated 1,4-dialdehyde structure to be the required structural motif for a low oral puncency thresholds and activation of the Transient Receptor Potential Channel A1 (TRPA1). Time-resolved NMR experiments confirmed the pungency mediating mechanism for electrophilic drimane sesquiterpene dialdehydes to be different from that found for other electrophilic pungent agents like isothiocyanates, which have been shown to undergo a covalent binding with cysteine residues in TRPA1. Instead, the high-impact chemosensates polygodial, warburganal, and 1ß-acetoxy-9-deoxy-isomuzigadial showed immediate reactivity with the ε-amino group of lysine side chains to give pyrrole-type conjugates, thus showing evidence for TRPA1 activation by covalent lysine modification.

Practical isolation of polygodial from Tasmannia lanceolata: a viable scaffold for synthesis.

Polygodial, a valuable sesquiterpene dialdehyde featuring an epimerizable stereocenter was efficiently extracted and isolated in gram-scale quantities (3.3% w/w) from Tasmannia lanceolata (Tasmanian native pepper) via a recently developed rapid pressurised hot water extraction (PHWE) technique that utilises an unmodified household espresso machine. This method was compared to the maceration of T. lanceolata under a range of conditions. Polygodial was used to achieve semi-syntheses of closely related sesquiterpene natural products drimendiol, (-)-drimenol, (+)-euryfuran, and some non-natural derivatives.

Antioxidant and α-amylase inhibitory activities of extract and isolates from Zygogynum pancheri subsp. arrhantum.

One new sesquiterpenoid (5R(*),8R(*),9R(*),10R(*))-cinnamolide (8), and seven known compounds, 5-hydroxy-7-methoxyflavonone (1), 8-hydroxy-3-(4'-hydroxyphenyl)-6,7-(2″,2″-dimethylchromene)-tetralone (2), 8-hydroxy-3-(3',4'-dihydroxyphenyl)-6,7-(2″,2″-dimethylchromene)-tetralone (3), 1ß-E-O-p-methoxycinnamoyl-bemadienolide (4), 1ß-O-(E-cinnamoyl)-6α-hydroxy-9-epi-polygodial (5), 1ß-O-(E-cinnamoyl)-6α-hydroxypolygodial (6), and 1ß-O-E-cinnamoylpolygodial (7) were isolated from the ethyl acetate extract of barks of Zygogynum pancheri subsp. arrhantum (Winteraceae). The structures of these molecules were assigned predominantly based on spectral data. The structure of compound 8 was confirmed by X-ray crystallographic analysis. Compounds 2 and 3 exhibited significant antioxidant activity, whereas compounds 1 and 4-7 showed significant α-amylase inhibitory activity.

Effect of packaging materials and storage on major volatile compounds in three Australian native herbs.

Lemon myrtle, anise myrtle, and Tasmanian pepper leaf are commercial Australian native herbs with a high volatile or essential oil content. Packaging of the herbs in high- or low-density polyethylene (HDPE and LDPE) has proven to be ineffective in preventing a significant loss of volatile components on storage. This study investigates and compares the effectiveness of alternate high-barrier property packaging materials, namely, polyvinylidene chloride coated polyethylene terephthalate/casted polypropylene (PVDC coated PET/CPP) and polyethylene terephthalate/polyethylene terephthalate/aluminum foil/linear low-density polyethylene (PET/PET/Foil/LLDPE), in prevention of volatile compound loss from the three native herbs stored at ambient temperature for 6 months. Concentrations of major volatiles were monitored using gas chromatography-mass spectrometry (GC-MS) techniques. After 6 months of storage, the greatest loss of volatiles from lemon myrtle was observed in traditional LDPE packaging (87% loss) followed by storage in PVDC coated PET/CPP (58% loss) and PET/PET/Foil/LLDPE (loss of 23%). The volatile loss from anise myrtle and Tasmanian pepper leaf stored in PVDC coated PET/CPP and PET/PET/Foil/LLDPE packaging was <30%. This study clearly indicates the importance of selecting the correct packaging material to retain the quality of herbs with high volatile content.

Growth inhibition of human colon carcinoma cells by sesquiterpenoids and tetralones of Zygogynum calothyrsum.

Bioassay-guided phytochemical investigation of Zygogynum calothyrsum using the human colon carcinoma cell lines COLO205 and KM12 led to the isolation of three new drimane-type sesquiterpenoids, 1ß-p-hydroxy-E-cinnamoyldrimeninol (1), 1ß-p-hydroxy-E-cinnamoyl-5α-hydroxydrimeninol (2), and methyl ether of 1ß-p-hydroxy-E-cinnamoyl-12α-methoxydrimeninol (3). Also isolated was the known 1ß-p-coumaroyloxypolygodial (4) together with two new tetralones, 3'-deoxyisozygolone A (5) and calothyrlone A (9), three known tetralones, isozygolone A (6), zygolone A (7), and 4'-O-methylzygolone A (8), and a known cinnamolide (10). Compounds 1, 7, and 8 demonstrated higher cytotoxicity against COLO205 (GI50 18, 17, and 11 µM, respectively) and KM12 (GI50 14, 14, and 17 µM, respectively) than the other compounds.

Composition of native Australian herbs polyphenolic-rich fractions and in vitro inhibitory activities against key enzymes relevant to metabolic syndrome.

Polyphenolic-rich fractions obtained from three native Australian herbs: Tasmannia pepper leaf, anise myrtle and lemon myrtle were characterised with regards to their composition, antioxidant capacities and inhibitory activities against α-glucosidase, pancreatic lipase and angiotensin I-converting enzyme, using in vitro models. Ellagic acid and derivatives were the dominant compounds of anise myrtle and lemon myrtle fractions, accompanied by flavonoids (catechin, myricetin, hesperetin, and quercetin). Tasmannia pepper leaf fraction comprised chlorogenic acid and quercetin derivatives, exhibited the highest oxygen radical absorbance capacity and effectively inhibited α-glucosidase (IC(50): 0.83 mg/ml) and pancreatic lipase (IC(50): 0.60 mg/ml). Anise myrtle and lemon myrtle fractions had pronounced α-glucosidase-inhibitory activities (IC(50): 0.30 and 0.13 mg/ml, respectively) and were less effective against lipase. Enzyme-inhibitory activities showed various levels of correlation with the levels of total phenolics and antioxidant capacities, indicating a specificity of individual phenolic compounds present in the isolated fractions to complex with proteins.

Influence of copigment derived from Tasmannia pepper leaf on Davidson's plum anthocyanins.

UNLABELLED: Davidson's plum (Davidsonia pruriens, F. Muell.), a native to Australian rainforests, large, crimson-red fruit, which superficially resembles plum, has been commercially cultivated in Australia since 1990s. The current production volume exceeds market demands therefore this study was designed to evaluate the suitability of Davidson's plum extract as a source of anthocyanin-based food colorant. The stability of the Davidson's plum extract towards heat treatment at 95 °C was higher than that of commercial mulberry colorant, but inferior to colorants derived from red cabbage and purple sweetpotato. An addition of a variety of phenolic acids significantly increased color intensity indicating the formation of copigmentation complexes. Commercial chlorogenic acid as well as extract from a native Australian herb rich in chlorogenic acid, Tasmannia pepper leaf (Tasmannia lanceolata, R. Br.), were both tested in model soft drink solutions subjected to light irradiation and heat treatment. In both cases, the addition of the copigment resulted in a lasting increase in color intensity. In conclusion, Davidson's plum extract can successfully be utilized as a source of natural food color. Extract from Tasmania pepper leaf can be used as a co-pigment for Davidson's plum anthocyanins. PRACTICAL APPLICATION: The color properties of an anthocyanin colorant derived from the native Australian fruit Davidson's plum are comparable to those of mulberry, which is currently applied as a food colorant in Australian food products. Utilization of Davidson's plum fruit as a source of natural color will allow the industry to increase the range of natural pigments and will create new opportunities for the emerging native food industry.

Chemical composition and antibacterial activity of the essential oil of Drimys granadensis L.f. leaves from Colombia.

Drimys granadensis L.f., a native plant from Colombia, has been included in the Vademecum de Plantas Medicinales de Colombia by the Instituto de Investigación de Recursos Biológicos Alexander von Humboldt, due to its widespread use in the folk medicine for the treatment of gastrointestinal ailments. The chemical composition of the essential oil of Drimys granadensis obtained by hydrodistillation of the leaves was analyzed by GC and GC/MS analyses. A total of 85 components were identified, with the major compounds germacrene D (1, 14.7%), sclarene (9.5%), a-cadinol (7.3%), longiborneol acetate (2, 6.3%), drimenol (4.2%), (Z)-ß-ocimene (3, 4.2%), a-pinene (4, 3.2%), and ß-elemene (5, 2.7%). The essential oil was also tested against eight bacteria using the Kirby-Bauer disk-diffusion method. Most of the Gram-positive bacteria tested were susceptible to the D. granadensis essential oil, while the Gram-negative bacteria tested were not.

Cytotoxic sesquiterpenoids from Winteraceae of Caledonian rainforest.

One secobutanolide, two butanolides and six drimane sesquiterpenoids were isolated from the bark and leaves of Zygogynum pancheri and Zygogynum acsmithii (Winteraceae) along with six known drimanes, isodrimanial, 1beta-O-p-methoxy-E-cinnamoyl-bemadienolide, 7-ketoisodrimenin, drimenin, polygodial and 1beta-E-cinnamoyl-6alpha-hydroxypolygodial. Their structures were elucidated through analysis of spectroscopic data. Drimane sesquiterpenoids with a dialdehyde function exhibited significant inhibitory activities in the in vitro cytotoxic assays against KB, HL60 and HCT116 cancer cell lines.

Biologically active tetralones from New Caledonian Zygogynum spp.

Bioassay guided purification of the ethyl acetate extracts of the bark and leaves of five New Caledonian Zygogynum species (Winteraceae) led to the isolation and characterization of four phenyl-3-tetralones (3,4-dihydronaphthalen-1(2H)-one). Their structures were determined by various NMR techniques and chemical studies. The absolute configuration of the compounds was established by circular dichroism. The compounds showed binding affinity for peroxisome proliferator-activated receptor-gamma (PPAR-gamma) and significant inhibitory activity against KB cancer cell line.

Sources of antioxidant activity in Australian native fruits. Identification and quantification of anthocyanins.

Selected native Australian fruits, muntries (Kunzea pomifera F. Muell., Myrtaceae), Tasmanian pepper berry (Tasmanian lanceolata R. Br., Winteraceae), Illawarra plum (Podocarpus elatus R. Br. ex Endl., Podocarpaceae), Burdekin plum (Pleiogynium timorense DC. Leenh, Anacardiaceae), Cedar Bay cherry (Eugenia carissoides F. Muell., Myrtaceae), Davidson's plum (Davidsonia pruriens F. Muell. var. pruriens, Davidsoniaceae), and Molucca raspberry (Rubus moluccanus var. austropacificus van Royen, Rosaceae), were evaluated as sources of antioxidants by 2,2-diphenyl-1-picrylhydrazyl and ferric reducing antioxidant power assays and compared with blueberry (Vaccinum spp. cv. Biloxi). The total reducing capacity of five fruits was 3.5-5.4-fold higher than that of blueberry, and the radical scavenging activities of muntries and Burdekin plum were 1.5- and 2.6-fold higher, respectively. The total phenolic level by Folin-Ciocalteu assay highly correlated with the antioxidant activity. Therefore, systematic research was undertaken to identify and characterize phenolic complexes. In the present study we report on the levels and composition of anthocyanins. The HPLC-DAD and HPLC/ESI-MS-MS (ESI = electrospray ionization) analyses revealed simple anthocyanin profiles of one to four individual pigments, with cyanidin as the dominating type. This is the first evaluation of selected native Australian fruits aiming at their utilization for the development of novel functional food products.

Pharmacological characterisation of the plant sesquiterpenes polygodial and drimanial as vanilloid receptor agonists.

This study was designed to assess the participation of transient receptor potential vanilloid 1 (TRPV1) in the biological effects induced by the plant-derived sesquiterpenes polygodial and drimanial. In rat isolated urinary bladder, polygodial and drimanial produced a tachykinin-mediated contraction that was inhibited by combination of NK(1) and NK(2) tachykinin receptor antagonists, SR 140333 and SR 48968. Furthermore, two different TRPV1 antagonists, capsazepine and ruthenium red prevented the contraction induced by both compounds. In addition, capsaicin, polygodial and drimanial displaced in a concentration-dependent manner the specific binding sites of [(3)H]-resiniferatoxin to rat spinal cord membranes, with a IC(50) values of 0.48, 4.2 and 3.2 microM, respectively. Likewise, capsaicin, polygodial and drimanial promoted an increase of [(45)Ca(2+)] uptake in rat spinal cord synaptosomes. In cultured rat trigeminal neurons, polygodial, drimanial and capsaicin were also able to significantly increase the intracellular Ca(2+) levels, effect that was significantly prevented by capsazepine. Together, the present results strongly suggest that the pharmacological actions of plant-derived sesquiterpenes polygodial and drimanial, seem to be partially mediated by activation of TRPV1. Additional investigations are needed to completely define the pharmacodynamic properties of these sesquiterpenes.

Protective effects of polygodial on gastric mucosal lesions induced by necrotizing agents in rats and the possible mechanisms of action.

The effects of polygodial isolated from the leaves of Tasmannia lanceolata on necrotizing agents-induced gastric lesions in rats were compared with capsaicin. Polygodial markedly inhibited the gastric mucosal lesions induced by several necrotizing agents, such as ethanol (ED(50)=0.029 mg/kg, p.o.), 0.6 M HCl (ED(50)=0.26 mg/kg, p.o.), and aspirin (ED(50)=0.38 mg/kg, p.o.), and partly inhibited the gastric mucosal lesions induced by indomethacin, but showed no significant effect on acid output in pylorus-ligated rats at doses of 0.05-0.5 mg/kg. The gastroprotection of polygodial was attenuated by pretreatment with indomethacin (10 mg/kg, s.c.), N(G)-nitro-L-arginine methyl ester (70 mg/kg, i.p.), N-ethylmaleimide (10 mg/kg, s.c.) and ruthenium red (3.5 mg/kg, s.c.). Polygodial (0.2 mg/kg, p.o.) increased the amount of reduced glutathione in gastric mucosa of ethanol-treated group. These results suggested that endogenous prostaglandins, nitric oxide, sulfhydryl compounds and vanilloid receptor-mediated effects are involved in the protective effect of polygodial.