Lychnophora Trichocarpha Research Articles

Eremantholide C from aerial parts of Lychnophora trichocarpha, as drug candidate: fraction absorbed prediction in humans and BCS permeability class determination.

Lychnophora trichocarpha (Spreng.) Spreng. ex Sch.Bip has been used in folk medicine to treat pain, inflammation, rheumatism and bruises. Eremantholide C, a sesquiterpene lactone, is one of the substances responsible for the anti-inflammatory and anti-hyperuricemic effects of L. trichocarpha. Considering the potential to become a drug for the treatment of inflammation and gouty arthritis, this study evaluated the permeability of eremantholide C using in situ intestinal perfusion in rats. From the permeability data, it was possible to predict the fraction absorbed of eremantholide C in humans and elucidate its oral absorption process. In situ intestinal perfusion studies were performed in the complete small intestine of rats using different concentrations of eremantholide C: 960μg/ml, 96μg/ml and 9.6μg/ml (with and without sodium azide), in order to verify the lack of dependence on the measured permeability as a function of the substance concentration in the perfusion solutions. Eremantholide C showed Peff values, in rats, greater than 5 × 10-5cm/s and fraction absorbed predicted for humans greater than 85%. These results indicated the high permeability for eremantholide C. Moreover, its permeation process occurs only by passive route, because there were no statistically significant differences between the Peff values for eremantholide C. The high permeability, in addition to the low solubility, indicated that eremantholide C is a biologically active substance BCS class II. The pharmacological activities, low toxicity and biopharmaceutics parameters demonstrate that eremantholide C has the necessary requirements for the development of a drug product, to be administered orally, with action on inflammation, hyperuricemia and gout.

Polymorphic characterization and implications on biopharmaceutics properties of potential anti-inflammatory drug candidate eremantholide C from Lychnophora trichocarpha (Brazilian Arnica)

To perform the polymorphic and physicochemical characterization of the potential anti-inflammatory drug, eremantholide C (EREC), as well as to evaluate the influence of these characteristics on its biopharmaceutics classification. Eremantholide C was obtained from chloroformic extract of Lychnophora trichocarpha and crystallized in two distinct solvents: chloroform (EREC 1) and ethyl acetate (EREC 2). To evaluate the polymorphism, EREC samples were submitted to melting point, purity, infrared spectroscopy, differential scanning calorimetry (DSC), X-ray powder diffraction, optical microscopy and scanning electron microscopy analysis. In addition, EREC samples crystallized after intrinsic dissolution study were submitted to DSC and X-ray powder diffraction analysis. EREC 1 showed fusion at 234.7-241.6°C, while EREC 2 showed fusion at 238.6-243.7°C. No polymorphic transitions were observed during the intrinsic dissolution experiment. A single sharp endothermic peak was obtained for the EREC samples. X-ray diffraction showed no crystallographic differences between the EREC samples. EREC 1 and EREC 2 showed birefringence under polarized light and indefinite morphology; however, the shape of the crystals was common to the two samples. Eremantholide C does not present classical or morphological polymorphism; therefore, there is no influence of crystalline transitions in the solubility and consequently in its biopharmaceutics classification and oral absorption process.

Development and validation of a stability indicating method for quantification of the sesquiterpene lactone eremantholide C from Lychnophora trichocarpha (Brazilian arnica)

Pharmacological activities as anti-inflammatory, anti-hyperuricemic, anti-gouty arthritis, antitumor and trypanocidal activities of the aerial parts from Lychnophora trichocarpha (Spreng.) Spreng. ex Sch.Bip., Asteraceae (Brazilian arnica) have already been proved. Eremantholide C is a sesquiterpene lactone and one of the active chemical constituents responsible for these activities presented by L. trichocarpha. Therefore, the aim of this work was to develop and validate a stability indicating HPLC method for eremantholide C. Eremantholide C stability was evaluated in L. trichocarpha ethanolic extract and in its isolated form. Analytical conditions employed C18 column, acetonitrile/water in gradient elution, flow of 0.8ml/min at 30°C. To correct for the loss of analyte during sample preparation the use of coumarin as an internal standard was necessary. The developed method provides good separation and resolution of the peaks, allowing quantification of eremantholide C, isolated or directly in the ethanolic extract, in internal standard presence. Validation results showed that this method is linear in the concentration range 2–180μg/ml, precise, accurate and specific. Stability studies showed that L. trichocarpha ethanolic extract and eremantholide C remain stable for 6 months when stored at room temperature and impermeable glass bottle, therefore they can be used safely and effectively within this period. While at 40°C there was stability loss, at 8°C a stability increase was observed for the extract and the isolated eremantholide C. Forced degradation studies showed that eremantholide C degraded under acidic and alkaline conditions and was stable for three days under neutral and oxidative conditions, and when exposed to high temperature. Thus, with the development of a stability indicative method and the application of it in eremantholide C stability studies, the results can guide the development of new products that adequately preserve the original features of the biologically active substance with quality, safety and efficacy.

Complete assignments of NMR data and assessment of trypanocidal activity of new eremantholide C derivatives.

Chemical transformations of eremantholide C (1), a sesquiterpene lactone that was isolated from Lychnophora trichocarpha Spreng. led to five new derivatives: 1',2'- epoxyeremantholide C (2), 5-n-propylamine-4,5-dihydro-1',2'-epoxyeremantholide C (3), 5-n-propylammonium-4,5-dihydro-1',2'-epoxyeremantholide C chloride (4), 5-n-propylammonium-4,5-dihydroeremantolide C chloride (5) and 16-O-ethyleremantholide C (6). The structures of all these derivatives were assigned on the basis of IR, MS, 1H and 13C NMR data by 1D and 2D techniques. Eremantholide C and the derivatives 2, 4 and 5 were evaluated against trypomastigotes Y and CL strains of Trypanosoma cruzi. Eremantholide C completely inhibited the growth of both the parasites strains while all derivatives were partially active against the CL strain and inactive against the Y strain.

Improvement of pharmacokinetic profile and efficacy of sesquiterpene lactone in nanocarriers in the experimental Chagas disease in mice

Chagas disease (CD), present in Latin America, is neglected and autochthone in other Continents [1]. Objetive: (i) evaluate the pharmacokinetic of LYC, a sesquiterpene lactone extracted from Lychnophora trichocarpha and LYC-NC formulated in polymeric nanocapsules (NC) and (ii) its efficacy in Swiss mice infected with Trypanosoma cruzi strains of different patterns of drugs resistance. NC was produced [2] at different concentrations of LYC and LYC-NCs were fully characterized, rending high percentages of encapsulation [3,4] and showing high stability. A new fast and precise bioanalytical method was developed (HPLC-UV detection), validated following FDA and National Agency of Sanitary Vigilance/Brazil guidelines [5] and further applied to LYC pharmacokinetic evaluation. It was observed an increasing in AUC (14 fold) and t1/2 (2,4 fold) and reduction of the Clearance (124 fold) of LYC when associated to PLA-PEG-NC x PLC-NC. The efficacy of free LYC, LYC-NC, BZ and controls was studied in mice inoculated with 1.0 × 104 trypomastigotes (acute phase) or 500 (chronic phase) of the CL, Y, Colombiana, VL10 T. cruzi strains. The cure rate in mice infected with all strains, acute and chronic phases, treated by IV/oral routes decreases in the following order: LYC-PLA-PEG-NC>LYC-PCL-NC>LYC free>BZ and was 0% for all controls. Cure rate of 88% was found with LYC-PLAPEG-NC treatment by oral route for resistant T. cruzi strain in chronic model (VL10 strain). This study is pioneer in the demonstration of better efficacy of LYC in vivo, when associated with NC, compared with BZ treatment of experimental CD, in both phases of infection with T. cruzi strains totally resistant to BZ/Nifurtimox. Furthermore, PLA-PEG-NC improved the pharmacokinetic parameters of LYC, increasing its plasma AUC and mean residence time, which contributes to the improvement of the efficacy in mice and showing the great potential of LYC for further clinical studies of CD treatment.

In Vitro Antitumor Activity of Sesquiterpene Lactones from Lychnophora trichocarpha

The sesquiterpene lactones lychnopholide and eremantholide C were isolated from Lychnophora trichocarpha Spreng. (Asteraceae), which is a plant species native to the Brazilian Savannah or Cerrado and popularly known as arnica. Sesquiterpene lactones are known to present a variety of biological activities including antitumor activity. The present paper reports on the evaluation of the in vitro antitumor activity of lychnopholide and eremantholide C, in the National Cancer Institute, USA (NCI, USA), against a panel of 52 human tumor cell lines of major human tumors derived from nine cancer types. Lychnopholide disclosed significant activity against 30 cell lines of seven cancer types with IC100 (total growth concentration inhibition) values between 0.41 µM and 2.82 µM. Eremantholide C showed significant activity against 30 cell lines of eight cancer types with IC100 values between 21.40 µM and 53.70 µM. Lychnopholide showed values of lethal concentration 50% (LC50) for 30 human tumor cell lines between 0.72 and 10.00 µM, whereas eremantholide C presented values of LC50 for 21 human tumor cell lines between 52.50 and 91.20 µM. Lychnopholide showed an interesting profile of antitumor activity. The α-methylene-γ-lactone present in the structure of lychnopholide, besides two α,β- unsaturated carbonyl groups, might be responsible for the better activity and higher cytotoxicity of this compound in relation to eremantholide C.

Toxicological evaluation of ethanolic extract of Lychnophora trichocarpha, Brazilian arnica

The species of the genus Lychnophora, Asteraceae, are popularly known as "arnica" and are native from Brazilian savana (Cerrado). They are widely used in Brazilian folk medicine as anti-inflammatory, to treat bruise, pain, rheumatism and for insect bites. For evaluation of acute toxicity, the ethanolic extract was given to albino female and male mice. In open-field test, the extract of Lychnophora trichocarpha (Spreng.) Spreng. (0.750 g/kg) induced a significant inhibition of the spontaneous locomotor activity and exploratory behavior of the animals were observed 1 and 4 h after administration. In traction test, the same dose reduced the muscular force 1 h after administration. The exploratory behavior reduced significantly in the group that received 0.50 g/kg, 1 and 4 h after administration of the extract. The animals that received the doses of 0.25, 0.50 and 0.75 g/kg did not show any change of blood biochemical parameters comparing to control group and showed some histopathological changes such as congestion and inflammation of kidney and liver. The dose of 1.5 g/kg caused the most serious signs of toxicity. Histopathological changes observed was hemorrhage in 62.5% and pulmonary congestion in 100% of the animals. Brain and liver congestion was found in 62.5% of the animals.

Pharmacological basis for use of Lychnophora trichocarpha in gouty arthritis: Anti-hyperuricemic and anti-inflammatory effects of its extract, fraction and constituents

Ethnopharmacological relevanceThe ethanolic extract of Lychnophora trichocarpha Spreng. is used in Brazilian folk medicine to treat bruise, pain and inflammatory diseases. Aim of the studyThe present study aimed at investigating whether ethanolic extract of L. trichocarpha, its ethyl acetate fraction and its main bioactive compounds could be useful to treat gouty arthritis by countering hyperuricemia and inflammation. Materials and methodsL. trichocarpha ethanolic extract (LTE), ethyl acetate fraction from ethanolic extract (LTA) and isolated compounds were evaluated for urate-lowering activity and liver xanthine oxidase (XOD) inhibition in oxonate-induced hyperuricemic mice. Anti-inflammatory activity in monosodium urate crystal-induced paw oedema, an experimental model of gouty arthritis, was also investigated. ResultsCrude ethanolic extract and its ethyl acetate fraction showed significant urate-lowering effects. LTE was also able to significantly inhibit liver xantine oxidase (XOD) activity in vivo at the dose of 250mg/kg. Luteolin, apigenin, lupeol, lychnopholide and eremantholide C showed the anti-hyperuricemic activities among tested compounds. Apigenin also showed XOD inhibitory activity in vivo. Luteolin, lychnopholide, lupeol and eremantholide C, in turn, did not shown significant inhibitory activity towards this enzyme, indicating that this mechanism is not likely to be involved in urate-lowering effects of those compounds. LTE, LTA, lupeol, β-sitosterol, lychnopholide, eremantholide, luteolin and apigenin were also found to inhibit monosodium urate crystals-induced paw oedema in mice. ConclusionsEthanolic extract of Lychnophora trichocarpha and some of its bioactive compounds may be promising agents for the treatment of gouty arthritis since they possesses both anti-hiperuricemic and anti-inflammatory properties.

Anti‐Inflammatory Sesquiterpene Lactones from Lychnophora trichocarpha Spreng. (Brazilian Arnica)

The aerial parts of Lychnophora trichocarpha Spreng. (Asteraceae) are used macerated in water or ethanol to treat inflammation, pain, rheumatism, contusions, bruises and insect bites in Brazilian traditional medicine. In this study, anti-inflammatory activity of ethanol extract from aerial parts of L. trichocarpha and its ethyl acetate fraction was investigated. Sesquiterpene lactones, lychnopholide (Lyc) and eremantholide C (EreC), isolated of ethyl acetate fraction, were also assayed for in vitro and in vivo anti-inflammatory activity. Topical treatment with ointments containing ethanol extract, its ethyl acetate fraction and sesquiterpene lactones significantly reduced carrageenan-induced mice paw oedema. In vitro assays demonstrated that Lyc inhibited interferon -γ/lipopolysaccharide -stimulated nitric oxide (NO) production in J774A.1 macrophages and increased production of IL-10 anti-inflammatory cytokine. The reduction of tumor necrosis factor-α (TNF-α) production by EreC was accompanied by an increased production of IL-10 in a concentration-dependent manner in J774A.1 macrophages. The anti-inflammatory effect of Lyc seems to involve the inhibition of production of NO and increased production of IL-10. The mechanism of the effect of EreC on the reduction of carrageenan-induced paw oedema may be attributed to inhibition of production of TNF-α and stimulation of IL-10 production. The results corroborate the use of ethanol extract from Lychnophora trichocarpha in folk medicine for anti-inflammatory action and indicate that the topical route is suitable for use.

Brine shrimp (Artemia salina Leach) bioassay of extracts from Lychnophoriopsis candelabrum and different Lychnophora species

The present study aimed to evaluate on Artemia salina the citotoxicity of twenty-two extracts from five species of the genus Lychnophora and one species of the genus Lychnophoriopsis. The extracts solubilized in DMSO and prepared at the final concentrations of 100, 250, 375, 500 and 600 μg mL-1 were added to tubes containing Artemia salina nauplii (10 units each) and filled to 5 mL total volume with artificial salt water. Lapachol and 5% DMSO were used as positive and negative controls, respectively. The samples were kept under light and dead larvae were counted after 24 hours of contact. LC50 was calculated by using Probit software. The crude ethanol extracts from five species showed low lethality in the following order: Lychnophora trichocarpha (LC50 = 672.38 μg mL-1) > Lychnophora pinaster (LC50 = 678.73 μg mL-1) > Lychnophora ericoides (LC50 = 738.09 μg mL-1) > Lychnophoriopsis candelabrum (LC50 = 812.57 μg mL-1) > Lychnophora passerina (LC50 = 921.78 μg mL-1). All tested extracts from L. candelabrum and chloroform extract from L. staavioides showed light toxicity on A. salina. Results indicated that there are substances with potential pharmacological activity in all tested species.

Complete assignments of 1H and 13C NMR data for trypanocidal eremantholide C oxide derivatives

The chemical transformations of eremantholide C (1), a trypanocidal sesquiterpene lactone isolated from Lychnophora trichocarpha Spreng., gave five new oxide derivatives: 3'-hydroxyeremantholide C (2), 1'-formyleremantholide C (3), 1'-carboxyeremantholide C (4), 1'-carbomethoxyeremantholide C (5) and sodium 1'-carboxylate of eremantholide C (6). The (1)H and (13)C NMR data of all these derivatives were assigned based on 1D and 2D techniques. The derivatives were evaluated against Y and CL strains of Trypanosoma cruzi. All of them were inactive against the Y strain. Compounds 2 and 5 displayed 100% activity on the CL strain while compounds 4 and 6 were partially active on the CL strain.

Antinociceptive and anti-inflammatory activities of ethanolic extracts of Lychnophora species

Extracts from Lychnophora species are traditionally used in Brazil as anti-inflammatory, and to treat bruise, pain and rheumatism. The ethanolic extract of aerial parts of five species of Lychnophoras and one specie of Lychnophoriopsis were examined for the antinociceptive (hot-plate and writhing tests) and anti-inflammatory (carrageenan-induced paw oedema test) activity in mice, by oral and topical routes, respectively. In the hot-plate test, the Lychnophora pinaster (0.75 g/kg) and Lychnophora ericoides (1.50 g/kg) extracts significantly increased the time for licking of the paws. The species Lychnophora passerina, Lychnophoriopsis candelabrum and Lychnophora pinaster, using the dose of 0.75 g/kg, and Lychnophora ericoides and Lychnophora trichocarpha in both doses evaluated (0.75 and 1.50 g/kg) significantly reduced the number of writhes induced by acetic acid. The administration of Lychnophora pinaster and Lychnophora trichocarpha ointments, in both concentrations evaluated (5 and 10%, w/w), and Lychnophora passerina and Lychnophoriopsis candelabrum, in the concentration of 10%, significantly reduced the paw oedema measured 3 h after carrageenan administration, suggesting, for the first time, an anti-inflammatory activity upon topical administration of these species. The present work comparatively demonstrated the antinociceptive and anti-inflammatory activities of some Brazilian Lychnophoras.

Xanthine oxidase inhibitory activity of Lychnophora species from Brazil (“Arnica”)

Twenty-two extracts from five Lychnophora species and one Lychnophoriopsis species, traditionally used in Brazil as analgesic, anti-inflammatory, and to treat bruise and rheumatism were examined for the inhibition of xanthine oxidase (XO), the enzyme that catalyses the metabolism of hypoxanthine and xanthine into uric acid. Sixteen extracts were tested. All of them were found to have excellent XO inhibitory activity, with inhibitions greater than 38% at 100 μg/mL in the assay mixture. The most active plants examined were Lychnophora trichocarpha, Lychnophora ericoides, Lychnophora staavioides and Lychnophoriopsis candelabrum, with inhibitions of 77%, 78%, 66% and 63% at 100 μg/mL, respectively, and IC 50 values of 6.16, 8.28, 33.97 and 37.70 μg/mL, respectively.