Efecto citotóxico de diterpenos derivados del ácido ent-kaureno aislados de Coespeletia moritziana (Sch. Bip. ex Wedd.) Cuatrec / Cytotoxic effect of ent-kaurene acid-derived diterpenes isolated from Coespeletia moritziana (Sch. Bip. Ex Wedd.) Cuatrec

The objective of the work was to study the cytotoxic effect of ent-kaurene acid derivatives obtained from Coespeletia moritziana (Sch. Bip. Ex Wedd.) Cuatrec., After analysis by GC/MS, IR and NMR. Isolating: kaurenic acid (I), grandifloric acid (II), 15-α-hydroxy kaurenic acid (III), 15 α-acetoxy-kaur 16-en-19-oic acid (IV), Kaurenol (V); and by hemisynthesis: 15,16-epoxy-17-acetoxy-kauran 19-oic acid (VI), 15-oxo-ent-kaur-16-en-19-oic acid (VIII), ester 2,3,4,6 -15-oxo-kaur-16-en-19-oic acid acetyl α-D-pyranosyl tetra-tetra (VII). Cytotoxicity was tested in human cancer cell lines: uterus (HeLa), lung (A-549), breast (MCF-7), African green monkey kidney non-tumor line (Vero) and human peripheral blood mononuclear cells (CMPS). Compound (I) was active against HeLa, A-549 and Vero. Compounds (II and VIII) showed moderate and good (IC50 ≤ 9 µM) cytotoxicity, respectively, against the five cell lines. Compound (V) showed moderate activity against A-549 and compound (VII), slight cytotoxicity against HeLa and A-549. Results that show the cytotoxic specificity of the isolated kaurenes and derivatives of Coespeletia moritzianaand their therapeutic potential.

El objetivo del trabajo fue estudiar el efecto citotóxico de derivados del ácido ent-kaureno obtenidos de Coespeletia moritziana (Sch. Bip. ex Wedd.) Cuatrec., previo análisis mediante GC/MS, IR y RMN. Aislandose: ácido kaurénico(I), ácido grandiflorénico (II), ácido 15-α-hidroxi kaurénico(III), ácido 15 α-acetoxi-kaur 16-en-19-oico (IV), Kaurenol (V); y por hemisíntesis: ácido 15,16-epoxi-17-acetoxi-kauran 19-oico (VI), ácido15-oxo-ent-kaur-16-en-19-oico (VIII), éster 2,3,4,6-tetra acetil α-D-piranosilo del ácido 15-oxo-kaur-16-en-19-oico (VII). La citotóxicidad fue ensayada en líneas celulares cancerosas humanas: útero (HeLa), pulmón(A-549), mama (MCF-7), línea no tumoral de riñón de mono verde africano (Vero) y células mononucleares humanas de sangre periférica (CMPS). El compuesto (I) resultó activo frente a HeLa, A-549 y Vero. Los compuestos (II y VIII), mostraron moderada y buena (IC50≤9µM) citotoxicidad respectivamente, frente a las cinco líneas celulares. El compuesto (V) presentó moderada actividad frente a A-549 y el (VII), leve citotoxicidad frente a HeLa y A-549. Resultados que evidencian la especificidad citotóxica de los kaurenos aislados y derivados de Coespeletia moritzianay su potencial terapéutico.

Astragalus species: Insights on its chemical composition toward pharmacological applications.

Astragalus L. is widely distributed throughout the temperate regions of Europe, Asia, and North America. The genus is widely used in folk medicine and in dietary supplements, as well as in cosmetics, teas, coffee, vegetable gums, and as forage for animals. The major phytoconstituents of Astragalus species with beneficial properties are saponins, flavonoids, and polysaccharides. Astragalus extracts and their isolated components exhibited promising in vitro and in vivo biological activities, including antiaging, antiinfective, cytoprotective, antiinflammatory, antioxidant, antitumor, antidiabesity, and immune-enhancing properties. Considering their proven therapeutic potential, the aim of this work is to give a comprehensive summary of the Astragalus spp. and their active components, in an attempt to provide new insight for further clinical development of these xenobiotics. This is the first review that briefly describes their ethnopharmacology, composition, biological, and toxicological properties.

Phytochemicals with Added Value from Morella and Myrica Species.

Terrestrial plants, due to their sessile nature, are highly exposed to environmental pressure and therefore need to produce very effective molecules that enable them to survive all the threats. Myrica and Morella (Myricaceae) are taxonomically close genera, which include species of trees or shrubs with edible fruits that exhibit relevant uses in traditional medicine. For instance, in Chinese or Japanese folk medicine, they are used to treat diarrhea, digestive problems, headache, burns, and skin diseases. A wide array of compounds isolated from different parts of Myrica and/or Morella species possess several biological activities, like anticancer, antidiabetic, anti-obesity, and cardio-/neuro-/hepatoprotective activities, both in vitro and in vivo, with myricanol, myricitrin, quercitrin, and betulin being the most promising. There are still many other compounds isolated from both genera whose biological activities have not been evaluated, which represents an excellent opportunity to discover new applications for those compounds and valorize Morella/Myrica species.

Cytotoxic diterpenes from roots of Crossopetalum gaumeri, a Celastraceae species from Yucatan Peninsula.

Four new diterpenes, crossogumerins A-D (1-4) along with six known ones (5-10) were isolated from the root bark of Crossopetalum gaumeri, an endemic medicinal plant from the Yucatan Peninsula. Their structures were elucidated on the basis of 1D and 2D NMR techniques, including HMQC, HMBC, and ROESY experiments. Compounds 1-5, 8-10 were evaluated for cytotoxicity against HeLa (carcinoma of the cervix) and Hep-2 (lung carcinoma) human tumor cells lines and against normal Vero cells (African green monkey kidney) in lag and log phase of growth. Podocarpane diterpenes, crossogumerin B (2) and nimbiol (10), exhibited the highest activity against HeLa cells (IC50 values of 3.1 and 8.1 µM, respectively), but also selectivity on Vero cells (SI 22.6 and 7.5, respectively). The preliminary SAR studies suggest that an epoxy moiety in ring B and a hydrogen bond-donor group strategically positioned in the diterpene core are important requirements for cytotoxicity and selectivity.

Chemical composition and biological activity of Conyza bonariensis essential oil collected in Mérida, Venezuela.

The essential oil from aerial parts of Conyza bonariensis (L) Cronquist collected in Mérida was obtained by hydrodistillation and analysed by GC/MS. The major components were trans-beta-farnesene (37.8%), trans-ocimene (20.7%) and beta-sesquiphellandrene (9.8%). Cytotoxicity assay was also performed with the essential oil against HeLa (cervix carcinoma), A-459 (lung carcinoma) and MCF-7 (breast adenocarcinoma) human cell lines and against normal Vero cells (African green monkey kidney) with IC50 values ranging from 1.4 to 45.8 microg/mL. Additionally, the essential oil presented a significant bactericidal effect against Bacillus cereus, while a moderate activity was observed against Staphylococcus epidermidis and Candida albicans.

Withaferin A-related steroids from Withania aristata exhibit potent antiproliferative activity by inducing apoptosis in human tumor cells.

Six new withanolides (1-6) along with eleven known ones (7-17) were isolated from the leaves of Withania aristata. Their structures were elucidated on the basis of spectroscopic analysis, including 1D and 2D NMR techniques. Semisynthesis of the minority metabolites 7 and 15 from compounds 6 and 9, respectively, as starting material, was performed. The isolated compounds as well as three derivatives (7a, 9a and 9b) of withaferin A were evaluated for cytotoxicity against HeLa (carcinoma of the cervix), A-549 (lung carcinoma) and MCF-7 (breast adenocarcinoma) human cancer cell lines, and against normal Vero cells (African green monkey kidney). Five compounds from this series (8, 9a, 9b, 11 and 13) exhibited potent antiproliferative effects on the tumor cells, even higher than the well known anticancer agent, withaferin A (9). Phosphatidylserine externalization, chromatin condensation, and caspase-3 activation clearly indicated apoptosis as a mechanism of action. The structure-activity relationship revealed valuable information on the pharmacophore for withanolide-type compounds.

A new natural spiro heterocyclic compound and the cytotoxic activity of the secondary metabolites from Juniperus brevifolia leaves.

A new natural spiro compound 3,4-dehydrotheaspirone and the known arctiol [1ß,6α-dihydroxy-4(14)-eudesmene] were isolated from Juniperus brevifolia. Arctiol is reported for the first time in the Juniperus genus. Their structures were established by 1D, and 2D NMR and MS spectra. Antimicrobial and cytotoxic activities of 1 and several secondary metabolites 3,4,5,6,7,8,9,10,11,12 previously isolated by our group from J. brevifolia were evaluated and some SAR has been established. The 18-hydroxydehydroabietane (4) displayed great antiproliferative activity against cancer cell lines tested, namely HeLa, A-549 and MCF-7. Compound 4 also presented a significant bactericidal effect against Bacillus cereus at different concentrations tested.

Antibacterial properties of phenolic triterpenoids against Staphylococcus epidermidis.

Two phenolic triterpenoids, pristimerol (30 µg/mL) and 8- EPI-6-deoxoblepharodol (20 µg/mL), obtained by catalytic reduction of pristimerin, exhibited bacteriostatic action against Staphylococcus epidermidis. This activity was not dependent on the inoculum size and the growth phase although it showed a stronger effect when cells were growing actively. Addition of phenolic triterpenoids to S. epidermidis cultures in the log-phase of growth led to an inhibitory effect on incorporation and uptake of radiolabeled precursors thymidine, uridine, leucine, and N-acetyl-glucosamine after 30 min of treatment. Furthermore, a clear release of UV-absorbing material and leakage of intracellular potassium were also detected. These findings, coupled with the high lipophilicity of these molecules, shown by high ClogP values, suggest that 8-EPI and pristimerol are able to interact within the lipid bilayer and as a consequence cause functional alterations on the cytoplasmic membrane of S. epidermidis cells.

Semisynthesis and biological evaluation of abietane-type diterpenes. Revision of the structure of rosmaquinone.

The new aromatic diterpenes 7beta-O-benzylrosmanol (3), 7beta-O-benzyl-11,12-di-O-methylrosmanol (4), and 7alpha-thiophenylcarnosic acid (5) have been obtained by partial synthesis from carnosol (1), an abundant natural diterpene present in Salvia species. The structures of these compounds were established from their physical and spectroscopic data. The known diterpenes sagequinone methide A (6), 7beta-O-methylrosmanol (7), 7-O-methylrosmanol (8), and rosmaquinone B (9) were obtained from rosmanol (2). The spectroscopic data of these semisynthetic diterpenes were identical to data reported in the literature. In addition, the new semisynthetic isorosmaquinone (10) was obtained from isorosmanol (12). The proton resonances of rosmaquinone (11) are reassigned based on 2D NMR spectroscopy. These compounds, as well as eight known analogues, were evaluated for cytotoxic and antimicrobial activities.

Cytotoxic activity of diterpenes and extracts of Juniperus brevifolia.

The hexane, dichloromethane, acetone and ethanol extracts of the leaves, bark and wood of Juniperus brevifolia were subjected to screening against human tumour cells lines from the cervix (HeLa) and larynx (Hep-2) in two different stages of the cell cycle. The dichloromethane and the chloroform-soluble portions of the acetone extracts of the leaves were active against both tumour cell lines. Chemical investigation of one of the most active extracts (dichloromethane extract from leaves) by preparative chromatography afforded compounds 1-10. Among these, compounds 1-3 were isolated for the first time from the Juniperus genus (18-hydroxyferruginol, 6,7-dehydrototarol and 7alpha-hydroxytotarol) while 4 and 5 are rare as natural products corresponding to (E)- and (Z)-isomers of 8alpha-hydroxy-labda-13(16),14-dien-19-yl-coumarate, respectively. The structures of all compounds were established based on various spectral data. The cytotoxic activity was evaluated for the first time for compounds 1-5 against HeLa and Vero cell lines in lag and log phases of growth. Compound 5 was shown to be active and selective to HeLa in the log phase.

Cytotoxic activity of lignans from Hibiscus cannabinus.

The antimicrobial and cytotoxic activities of six lignans isolated from the core and bark acetone extracts of Hibiscus cannabinus have been investigated. Two compounds (2 and 3) showed strong cytotoxic activity against HeLa, Hep-2 and A-549 cell lines while compound 5 showed moderate activity on HeLa cells when they were in advanced stage of cellular division. The compounds did not exhibit antimicrobial activity.

Antimicrobial activity of 6-oxophenolic triterpenoids. Mode of action against Bacillus subtilis.

Zeylasteral and demethylzeylasteral are 6-oxophenolic triterpenoids isolated from the root of Maytenus blepharodes, which have antimicrobial activity against Gram-positive bacteria and the yeast Candida albicans. The time-kill curves for zeylasteral and demethylzeylasteral at concentrations twice their MICs, against Bacillus subtilis showed that the colony forming units were reduced in 3-log10 and > 4-log10 respectively. This reduction was dependent on inoculum size and the growth phase of cells, and was greater when the compounds were incorporated in the exponential phase, indicating a bacteriolytic effect. Treatment with both agents, particularly with zeylasteral (20 microg/mL) caused a reduction of optical density at 550 nm. With regard to the synthesis of DNA, RNA, protein and cell wall, the compounds exhibited the fastest inhibition against cell wall synthesis. Thus, the predisposition to lysis, the morphological changes seen by microscopy, and the complete inhibition in the incorporation the N-acetyl-d-[1 - 14C]glucosamine, suggest that the phenolic compounds compromise the cell wall synthesis and/or cytoplasmic membrane.

Antimicrobial terpenoids from the oleoresin of the Peruvian medicinal plant Copaifera paupera.

Twelve known diterpenes 1 - 11 and 13, and three known sesquiterpenes 14 - 16, along with a new C(20) - C(15) terpenoid 17, with a structure based on an unprecedented skeleton in which a labdane diterpene is linked to a monocyclic sesquiterpene by an ester bridge, were isolated from the oleoresin of the Peruvian medicinal plant Copaifera paupera (Herzog) Dwyer (Leguminosae). Their structures were elucidated on the basis of spectral analysis, including homo- and heteronuclear correlation NMR experiments (COSY, ROESY, HMQC and HMBC), and by comparison with data in the literature. The leishmanicidal, antimicrobial, cytotoxic, and aldose reductase inhibitory activities were studied. Compounds 1 and 11 showed significant antimicrobial activity (MIC < 10 microg/ml) against Gram-positive bacteria, comparable with cephotaxime used as control. Compound 2 exhibited moderate cytotoxic activity against four cancer cell lines.