New Acetophenones and Chromenes from the Leaves of Melicope barbigera A. Gray.

The dichloromethane extract from leaves of Melicope barbigera (Rutaceae), endemic to the Hawaiian island of Kaua'i, yielded four new and three previously known acetophenones and 2H-chromenes, all found for the first time in M. barbigera. The structures of the new compounds obtained from the dichloromethane extract after purification by chromatographic methods were unambiguously elucidated by spectroscopic analyses including 1D/2D NMR spectroscopy and HRESIMS. The absolute configuration was determined by modified Mosher's method. Compounds 2, 4 and the mixture of 6 and 7 exhibited moderate cytotoxic activities against the human ovarian cancer cell line A2780 with IC50 values of 30.0 and 75.7 µM for 2 and 4, respectively, in a nuclear shrinkage cytotoxicity assay.

Abyssinone V, a prenylated flavonoid isolated from the stem bark of Erythrina melanacantha increases oxidative stress and decreases stress resistance in Caenorhabditis elegans.

OBJECTIVES: Recent studies showed that distinct extracts of Erythrina species used in the traditional medicine of sub-Saharan Africa are protective against stress conditions. However, the underlying molecular mechanisms as well as relevant compounds remain unclear. METHODS: We used the model organism Caenorhabditis elegans to investigate compounds isolated from the stem bark of Erythrina melanacantha (abyssinone V (1), abyssinon-4'O-methylether (2), sigmoidin B-4'O-methylether (3), glabranin (4), 8-prenylnaringenin (5), citflavanone (6), exiguaflavanone (7) and homoeriodictyol (8)). Antioxidative capacity in vitro (trolox equivalent antioxidative capacity assay) and modulation of oxidative stress in vivo (2', 7'-dichlorofluorescein assay) were investigated; stress resistance was analysed using the nucleic acid stain SYTOX green. KEY FINDINGS: None of the prenylated flavonoids caused protection against thermal stress; in contrast, most of the compounds (1, 4, 5, 8) decreased stress resistance. None of the compounds decreased the accumulation of reactive oxygen species, but abyssinone V (1) caused an increase in oxidative stress. In line with these results, none of these compounds showed radical-scavenging effects in vitro. CONCLUSIONS: The stem bark of E. melanacantha contains various prenylated flavonoids, but no compound protected C. elegans against stress conditions. In contrast, abyssinone V increases oxidative stress and reduces stress resistance in this model organism.

Insecticidal activity of the essential oil of Ligusticum mutellina roots.

The essential oil obtained from roots of different collections of Ligusticum mutellina was tested against 3rd instar armyworms, Pseudaletia unipuncta (Lepidoptera: Noctuidae), for insecticidal activity. The main compounds were isolated and their structures were elucidated using 2D-NMR techniques. Our collections contained dillapiole, ligustilide and myristicin as major compounds. The previously reported sarisan was not present, moreover its occurrence in L. mutellina should be revised based on our findings.

Prenylated flavanone derivatives isolated from Erythrina addisoniae are potent inducers of apoptotic cell death.

Extracts of Erythrina addisoniae are frequently used in the traditional medicine of Western Africa, but insufficient information about active compounds is available. From the stem bark of E. addisoniae, three (1, 2, 4) and three known (3, 5, 6) flavanones were isolated: addisoniaflavanones I and II, containing either a 2″,3″-epoxyprenyl moiety (1) or a 2″,3″-dihydroxyprenyl moiety (2) were shown to be highly toxic (MTT assay: EC50 values of 5.25±0.7 and 8.5±1.3 µM, respectively) to H4IIE hepatoma cells. The cytotoxic potential of the other isolated flavanones was weaker (range of EC50 values between 15 and >100 µM). Toxic effects of addisoniaflavanone I and II were detectable after 3h (MTT assay). Both compounds induced an apoptotic cell death (caspase-3/7 activation, nuclear fragmentation) in the hepatoma cells and, at high concentrations, also necrosis (membrane disruption: ethidium bromide staining). Formation of DNA strand breaks was not detectable after incubation with these compounds (comet assay). In conclusion, the prenylated flavanones addisoniaflavanones I and II may be of interest for pharmacological purposes due to their high cytotoxic and pro-apoptotic potential against hepatoma cells.

Lobatin B inhibits NPM/ALK and NF-κB attenuating anaplastic-large-cell-lymphomagenesis and lymphendothelial tumour intravasation.

An apolar extract of the traditional medicinal plant Neurolaena lobata inhibited the expression of the NPM/ALK chimera, which is causal for the majority of anaplastic large cell lymphomas (ALCLs). Therefore, an active principle of the extract, the furanoheliangolide sesquiterpene lactone lobatin B, was isolated and tested regarding the inhibition of ALCL expansion and tumour cell intravasation through the lymphendothelium. ALCL cell lines, HL-60 cells and PBMCs were treated with plant compounds and the ALK inhibitor TAE-684 to measure mitochondrial activity, proliferation and cell cycle progression and to correlate the results with protein- and mRNA-expression of selected gene products. Several endpoints indicative for cell death were analysed after lobatin B treatment. Tumour cell intravasation through lymphendothelial monolayers was measured and potential causal mechanisms were investigated analysing NF-κB- and cytochrome P450 activity, and 12(S)-HETE production. Lobatin B inhibited the expression of NPM/ALK, JunB and PDGF-Rß, and attenuated proliferation of ALCL cells by arresting them in late M phase. Mitochondrial activity remained largely unaffected upon lobatin B treatment. Nevertheless, caspase 3 became activated in ALCL cells. Also HL-60 cell proliferation was attenuated whereas PBMCs of healthy donors were not affected by lobatin B. Additionally, tumour cell intravasation, which partly depends on NF-κB, was significantly suppressed by lobatin B most likely due to its NF-κB-inhibitory property. Lobatin B, which was isolated from a plant used in ethnomedicine, targets malignant cells by at least two properties: I) inhibition of NPM/ALK, thereby providing high specificity in combating this most prevalent fusion protein occurring in ALCL; II) inhibition of NF-κB, thereby not affecting normal cells with low constitutive NF-κB activity. This property also inhibits tumour cell intravasation into the lymphatic system and may provide an option to manage this early step of metastatic progression.

The germacranolide sesquiterpene lactone neurolenin B of the medicinal plant Neurolaena lobata (L.) R.Br. ex Cass inhibits NPM/ALK-driven cell expansion and NF-κB-driven tumour intravasation.

BACKGROUND: The t(2;5)(p23;q35) chromosomal translocation results in the expression of the fusion protein NPM/ALK that when expressed in T-lymphocytes gives rise to anaplastic large cell lymphomas (ALCL). In search of new therapy options the dichloromethane extract of the ethnomedicinal plant Neurolaena lobata (L.) R.Br. ex Cass was shown to inhibit NPM/ALK expression. PURPOSE: Therefore, we analysed whether the active principles that were recently isolated and found to inhibit inflammatory responses specifically inhibit growth of NPM/ALK+ ALCL, leukaemia and breast cancer cells, but not of normal cells, and the intravasation through the lymphendothelial barrier. METHODS: ALCL, leukaemia and breast cancer cells, and normal peripheral blood mononuclear cells (PBMCs) were treated with isolated sesquiterpene lactones and analysed for cell cycle progression, proliferation, mitochondrial activity, apoptosis, protein and mRNA expression, NF-κB and cytochrome P450 activity, 12(S)-HETE production and lymphendothelial intravasation. RESULTS: In vitro treatment of ALCL by neurolenin B suppressed NPM/ALK, JunB and PDGF-Rß expression, inhibited the growth of ALCL cells late in M phase, and induced apoptosis via caspase 3 without compromising mitochondrial activity (as a measure of general exogenic toxicity). Moreover, neurolenin B attenuated tumour spheroid intravasation probably through inhibition of NF-κB and CYP1A1. CONCLUSION: Neurolenin B specifically decreased pro-carcinogenic NPM/ALK expression in ALK+ ALCL cells and, via the inhibition of NF-kB signalling, attenuated tumour intra/extravasation into the lymphatics. Hence, neurolenin B may open new options to treat ALCL and to manage early metastatic processes to which no other therapies exist.

Metabolism of thymol and trans-anethole in larvae of Spodoptera litura and Trichoplusia ni (Lepidoptera: Noctuidae).

Metabolism of the monoterpenoid thymol and the phenylpropanoid trans-anethole, both constituents of essential oils, was investigated following topical and oral administration of the compounds to the tobacco cutworm (Spodoptera litura) and the cabbage looper (Trichoplusia ni). In both species and irrespective of route, administration of thymol resulted in the excretion of its 3-O-beta-glucoside, whereas trans-anethole was hydroxylated on the side chain methyl group. Both metabolites were isolated and their structures elucidated by interpretation of their mass and NMR spectra. Previous experiments indicated that trans-anethole synergized the toxicity of thymol in S. litura, but analyses of feces indicated that metabolism of thymol was not significantly suppressed when the two compounds were orally coadministered to T. ni larvae.

New diacylated 2-hydroxythymol derivatives from Melampodium divaricatum.

Melampodium divaricatum is a medicinal plant, which occurs in Central America. In a recent paper we reported the occurrence of acylated 2-hydroxy thymol glycosides as main constituents in this plant. This paper deals with the isolation of two new 2,5-dihydroxythymol ester derivatives. The formerly reported sesquiterpene lactone mikanokryptin was not found in our plant material.

Antiplasmodial activities of sesquiterpene lactones from Eupatorium semialatum.

Eupatorium semialatum, Plasmodium falciparum, Sesquiterpene Lactones Eupatorium semialatum is a member of the Asteraceae, which occurs in Guatemala. Previously, we reported the occurrence of sesquiterpene lactones of the eudesmanolide type as main constituents in the leaves. This paper deals with the isolation and identification of the first guaianolide found in E. semialatum. Since this plant is used against malaria and other diseases in the Guatemalan folk medicine, the main sesquiterpene lactones were tested for their activities against Plasmodium falciparum in vitro.

Prenylated pterocarpanes from Erythrina melanacantha.

The stem bark of Erythrina melanacantha ssp. melanacantha Taub. ex Harms. afforded one new and five known prenylated pterocarpanes (phaseollin, isoneorautenol, erybraedin A, phaseollidin, and homoedudiol). The new compound was identified as 2',3'-epoxy-homoedudiol [3-hydroxy-8-(3,3-dimethyl-oxiranyl-methyl)pterocarpane]. All structures were elucidated by interpretation of their 1D and 2D NMR spectra.

Pseudoguaianolide sesquiterpene lactones with high activities against the human malaria parasite Plasmodium falciparum.

Four sesquiterpene lactones of the pseudo-guaianolide type (helenalin, dihydrohelenalin and their acetates), have shown activities against asexual blood forms of Plasmodium falciparum in vitro. Their IC(50) values were situated in the range of 0.23 to 7.41 micro M. The activity found for helenalin was high, even compared to the activity found for artemisinin.