Bradley S. Simpson

In Vivo Activity of Benzoyl Ester Clerodane Diterpenoid Derivatives from Dodonaea polyandra

Four new benzoyl ester clerodane diterpenoids, 15,16-epoxy-8α-(benzoyloxy)methylcleroda-3,13(16),14-trien-18-oic acid (1), 15,16-epoxy-8α-(benzoyloxy)methyl-2α-hydroxycleroda-3,13(16),14-trien-18-oic acid (2), 15,16-epoxy-8α-(benzoyloxy)methyl-2-oxocleroda-3,13(16),14-trien-18-oic acid (3), and 15,16-epoxy-2α-benzoyloxycleroda-3,13(16),14-trien-18-oic acid (4), have been isolated from the leaves and stems of Dodonaea polyandra. The anti-inflammatory activities of compounds 1, 2, and 4 were evaluated by means of 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse ear edema. Compounds 2 and 4 exhibited maximum inhibition of inflammation (70-76%) at doses of 0.22 and 0.9 μmol/ear, respectively. Modest activity (~45% inhibition) was maintained at nanomole/ear doses.

Rare, seven-membered cyclic ether labdane diterpenoid from Dodonaea polyandra

Previous phytochemical studies on the leaf resin of dioecious plant species Dodonaea polyandra have identified the presence of furanoclerodane diterpenoids. As part of ongoing research on this species the chemical profile of an individual plant displaying male flowers was investigated. Repeated chromatographic separation of a resinous extract from the leaves of the plant yielded three labdane diterpenoids, 13,17-epoxy-13-methyl-15-oxo-labda-7-ene (1), 17-hydroxy-13-methyl-labda-7,13Z-diene-15-oic acid (2) and 13-methyl-17-oxo-labda-7,13Z-diene-15-oic acid (3) and a fourth known labdane diterpenoid (4) reported as being isolated from a natural source for the first time. Structural elucidation was carried out using conventional 1D and 2D NMR and mass spectrometry together with other complementary techniques (UV and IR). The leaf extract from this individual of D. polyandra with male flowers present displays a marked difference in the chemical composition of diterpenoids compared to previously studied extracts from the leaves of this species.

Dioecy in plants--is it an important factor for phytochemists to consider?

Plants with separate male and female flowers are termed dioecious. Dioecy is not rare, yet is a characteristic that could potentially impact the phytochemical and subsequent pharmacological properties of a species. This is a brief insight which highlights why the sex of the plant might be an important factor to consider for researchers within phytochemistry related fields.

Biological activity and LC-MS/MS profiling of extracts from the Australian medicinal plant Acacia ligulata (Fabaceae)

Abstract Acacia ligulata A.Cunn. ex Benth. (Fabaceae: Mimosoideae) is a native Australian plant used traditionally by Australian Aboriginal groups. This study was undertaken to investigate the bioactivity of A. ligulata extracts and to evaluate their chemical composition. Potential antibacterial, cytotoxic and enzyme inhibitory effects relevant to traditional medicinal and food uses of the species were examined and LC-MS/MS was performed to investigate the chemical composition. Antibacterial activity was observed for bark and leaf extracts with an MIC for the bark extract of 62.5 μg/mL against Streptococcus pyogenes. Pod extracts showed cytotoxic effects against cancer cells, with the highest activity against melanoma SK-MEL28 cells with IC50 values between 40.8 and 80.6 μg/mL. Further, the leaf and pod extracts also inhibited α-amylase EC-3.2.1.1 and α-glucosidase EC-3.2.1.20 with IC50 values between 9.7–34.8 and 12.6–64.3 μg/mL, respectively. The LC-MS/MS profiling indicated that several different saponins were present in the active extracts.

Arid awakening: new opportunities for Australian plant natural product research

The importance of plants and other natural reserves as sources for biologically important compounds, particularly for application in food and medicine, is undeniable. Herein we provide a historical context of the major scientific research programs conducted in Australia that have been aimed at discovering novel bioactive natural products from terrestrial plants. Generally speaking, the main approaches used to guide the discovery of novel bioactive compounds from natural resources have included random, ethnobotanical and chemotaxonomic strategies. Previous Australian plant natural product research campaigns appear to have lacked the use of a fourth strategy with equally high potential, namely the ecologically guided approach. In addition, many large studies have sampled plant material predominantly from tropical regions of Australia, even though arid and semi-arid zones make up 70% of mainland Australia. Therefore, plants growing in arid zone environments, which are exposed to different external stressors (e.g. low rainfall, high ultraviolet exposure) compared with tropical flora, remain an untapped reservoir of potentially novel bioactive compounds. Research of Australian arid zone plants that is ecologically guided creates a new opportunity for the discovery of novel bioactive compounds from plants (and potentially other biota) for application in health care, food and agricultural industries.

Antiproliferative Aporphine Alkaloids from Litsea glutinosa and Ethnopharmacological Relevance to Kuuku I’yu Traditional Medicine

Australian Aboriginal people have a long history of relying on plants for the treatment of various ailments and illnesses. Our ongoing collaborative research project initiated by Chuulangun Aboriginal Corporation (Cape York, Australia) has recently focussed on revealing whether Kuuku I’yu plant medicines possess anticancer-related activities and the chemistry responsible for this. Here, we present results from a study of the plant Litsea glutinosa, used traditionally for the treatment of gastrointestinal disorders. Four known aporphine alkaloids N-methylactinodaphnine (1), boldine (2), N-methyllaurotetanine (3), and isoboldine (4) were isolated by activity-guided fractionation and tested for cytotoxicity against HT29, SKMEL28, and primary human keratinocytes. Compound 1 was the most cytotoxic and this observation may be explained by the presence of a 1,2-methylenedioxy group. In silico docking revealed that a plausible mechanism for the observed cytotoxicity is the stabilization of a topoisomerase II (β) DNA–enzyme complex. The ethnopharmacological relevance of this study is discussed in the context of researching and using traditional knowledge in biomolecular discovery.

Isolation and Structural Characterization of Echinocystic Acid Triterpenoid Saponins from the Australian Medicinal and Food Plant Acacia ligulata

The Australian plant Acacia ligulata has a number of traditional food and medicinal uses by Australian Aboriginal people, although no bioactive compounds have previously been isolated from this species. Bioassay-guided fractionation of an ethanolic extract of the mature pods of A. ligulata led to the isolation of the two new echinocystic acid triterpenoid saponins, ligulatasides A (1) and B (2), which differ in the fine structure of their glycan substituents. Their structures were elucidated on the basis of 1D and 2D NMR, GC-MS, LC-MS/MS, and saccharide linkage analysis. These are the first isolated compounds from A. ligulata and the first fully elucidated structures of triterpenoid saponins from Acacia sensu stricto having echinocystic acid reported as the aglycone. Compounds 1 and 2 were evaluated for cytotoxic activity against a human melanoma cancer cell line (SK-MEL28) and a diploid fibroblast cell line (HFF), but showed only weak activity.

Quantification of salivary cortisol from captive dingoes (Canis dingo) in relation to age, sex, and breeding season: implications for captive management

Captive-breeding programs can play a key role in the conservation of threatened species such as the Australian dingo (Canis dingo). It is important to determine whether holding and rearing practices impose stressors that impact negatively on program outcomes and the health and wellbeing of the captive population. Despite evidence that chronic stress has significant welfare implications, our understanding of ‘stress’ in either wild or captive dingoes remains limited. In a first attempt to rectify this, we report salivary cortisol concentrations in juvenile and adult dingoes held in a captive colony. Dingo puppies (n = 8, M = 0.484 ± 0.09 μg dL–1) were found to have higher concentrations than adults (n = 12, M = 0.106 ± 0.031 μg dL–1) (P < 0.0001). Concentrations in adult females (n = 6, M = 0.113 ± 0.030 μg dL–1) and males (n = 6, M = 0.099 ± 0.033 μg dL–1) did not significantly differ (P = 0.4740). Our preliminary findings also suggest that during the annual breeding season, males (but not females) have elevated levels of corticosteroids. Establishing a reference range for cortisol concentrations is vital for researchers and wildlife carers attempting to measure stressors in both captive and wild dingo populations. This study provides useful insight into the influence of time of day, development, and seasonality on cortisol concentrations. Suggestions for future research and implications of routine cortisol evaluation to aid better management practices are also discussed.

In vitro metabolism of the anti-inflammatory clerodane diterpenoid polyandric acid A and its hydrolysis product by human liver microsomes and recombinant cytochrome P450 and UDP-glucuronosyltransferase enzymes

Abstract 1. The metabolism of the anti-inflammatory diterpenoid polyandric acid A (PAA), a constituent of the Australian Aboriginal medicinal plant Dodonaea polyandra, and its de-esterified alcohol metabolite, hydrolysed polyandric acid A (PAAH) was studied in vitro using human liver microsomes (HLM) and recombinant UDP-glucuronosyltransferase (UGT) and cytochrome P450 (CYP) enzymes. 2. Hydrolysis of PAA to yield PAAH occurred upon incubation with HLM. Further incubations of PAAH with HLM in the presence of UGT and CYP cofactors resulted in significant depletion, with UGT-mediated depletion as the major pathway. 3. Reaction phenotyping utilising selective enzyme inhibitors and recombinant human UGT and CYP enzymes revealed UGT2B7 and UGT1A1, and CYP2C9 and CYP3A4 as the major enzymes involved in the metabolism of PAAH. 4. Analysis of incubations of PAAH with UDP-glucuronic acid-supplemented HLM and recombinant enzymes by UPLC/MS/MS identified three glucuronide metabolites. The metabolites were further characterised by β-glucuronidase and mild alkaline hydrolysis. The acyl glucuronide of PAAH was shown to be the major metabolite. 5. This study demonstrates the in vitro metabolism of PAA and PAAH and represents the first systematic study of the metabolism of an active constituent of an Australian Aboriginal medicinal plant.