RESUMEN

The flowers of the species of Malpighiaceae in the Neotropical Region are relatively uniform in their morphology due to their dependence on oil-collecting bees as their main pollinators. However, many species of the genus Galphimia seem to have acquired a different floral syndrome, lacking markedly zygomorphic flowers and developed elaiophores in the calyx. Likewise, these species present anthers with great development, probably in response to the selection of pollinators that collect pollen. Galphimia australis incorporated some of these traits but also retained some residual characteristics typical of species pollinated by oil bees. This leads to many questions on how these flowers ensure their pollination. Inquiring about the reduction or modification of these characteristics allows us to understand how G. australis achieves a different pollination syndrome. In this research, we carry out a detailed morphological and anatomical study of the flowers and pollen grain devolvement of G. australis and floral visitors were observed and captured. Results were analyzed in order to determine how this species changed from the oil-floral syndrome, typical of neotropical Malpighiaceae, to one syndrome with pollen as the main reward.

Asunto(s)

Galphimia , Malpighiaceae , Animales , Abejas , Polinización/fisiología , Malpighiaceae/fisiología , Flores/anatomía & histología , Polen/fisiología

RESUMEN

The anti-depressive and anxiolytic effect of galphimine B (isolated from Galphimia glauca) has been demonstrated by researchers. Therefore, it is necessary to explore extraction techniques that produce materials with adequate quality for pharmaceutical applications. In this work, supercritical extractions of galphimines from Galphimia glauca were performed in the presence of carbon dioxide. Pressure, temperature, particle diameter, and flow rate effects were examined to explore the conditions with the highest yield and the concentration profile of galphimines in the studied interval. The identification of the nor-seco triterpenoids and galphimine B and E was carried out by HPLC analyses. The mathematical modeling of the extraction curves was attained by the approaches proposed by Sovová and Papamichail et al. According to results, the highest yield 2.22% was obtained at 323.15 K, 326 µm, 3 L/min, and 33.75 MPa. Meanwhile, the content of galphimine B in the extract was, on average, 19.5 mg·g-1.

Asunto(s)

Dióxido de Carbono/química , Cromatografía con Fluido Supercrítico/métodos , Galphimia/química , Extractos Vegetales/química , Cromatografía Líquida de Alta Presión , Triterpenos/química

RESUMEN

Both DNA barcoding and phylogenetic data of the studied botanical material suggested the existence a new population of Galphimia glauca. Their leaves afforded three new nor-3,4-seco-friedelanes named galphimines M-O, together with known galphimines D, E, G, and I. Galphimines M and N possess bicyclic orthoacetates which are the first examples of orthoesters found in the Malpighiaceae family, while galphimine O has a 27,20-δ-lactone moiety. The structures elucidation followed from spectroscopic means and the absolute configuration followed from single crystal X-ray diffraction analyses. Tests for antibacterial and antifungal activities of galphimines N and M showed no promising effects.

Asunto(s)

Galphimia/química , Triterpenos/química , Cristalografía por Rayos X , Modelos Moleculares , Conformación Molecular , Triterpenos/aislamiento & purificación

RESUMEN

Galphimia glauca is a plant that is endemic to Mexico and has been commonly used since pre-Hispanic times to treat various illnesses, including central nervous system disorders and inflammation. The first studies investigating a natural population of G. glauca in Mexico showed that the plant has anxiolytic and sedative activities in mice and humans. The plant's bioactive compounds were isolated and identified, and they belong to a family of nor-secofriedelanes called galphimines. The integration of DNA barcoding and thin-layer chromatography analysis was performed to clarify whether the botanical classification of the populations in the study, which were collected in different regions of Mexico, as G. glauca was correct or if the populations consist of more than one species of the genus Galphimia. We employed six DNA barcodes (matK, rbcL, rpoC1, psbA-trnH, ITS1 and ITS2) that were analyzed individually and in combination and then compared each other, to indicate differences among the studied populations. In the phylogenetic analysis, ITS1 and ITS2 markers as well as the combination of all DNA regions were the most efficient for discriminating the population studied. The thin-layer chromatography analysis exhibited four principal chemical profiles, one of which corresponded to the populations that produced galphimines. DNA barcoding was consistent and enabled us to differentiate the populations that produce galphimines from those that do not. The results of this investigation suggest that the studied populations belong to at least four different species of the genus Galphimia. The phylogenetic analysis and the thin-layer chromatography chemical profiles were convenient tools for establishing a strong relationship between the genotype and phenotype of the studied populations and could be used for quality control purposes to prepare herbal medicines from plants of the genus Galphimia.

Asunto(s)

Galphimia/clasificación , Plantas Medicinales/clasificación , Animales , Ansiolíticos/aislamiento & purificación , Secuencia de Bases , Cromatografía en Capa Delgada , Código de Barras del ADN Taxonómico , ADN de Plantas/genética , Galphimia/química , Galphimia/genética , Genes de Plantas , Humanos , Hipnóticos y Sedantes/aislamiento & purificación , México , Ratones , Filogenia , Plantas Medicinales/química , Plantas Medicinales/genética , Especificidad de la Especie

RESUMEN

Galphimine-B (G-B), a compound isolated from Galphimia glauca, has been shown to possess important anxiolytic activity. In this study, we evaluated the effectiveness and tolerability of a G-B standardized extract (experimental treatment) that was administered daily for 10 weeks in patients with moderate or severe Generalized Anxiety Disorder (GAD). Alprazolam was used as control treatment and administered under the same conditions. A total of 167 patients were included. At the start of the study, the severe anxiety condition prevailed, with an average on the Hamilton Anxiety Scale of 35.1 ± 8.8 and 35.8 ± 8.1 points in the control and experimental groups, respectively. After the 10 weeks of administration, the average was reduced in the control group to 4.6 ± 6.5 points and in the experimental group to 3.5 ± 5.5 points. Therapeutic success in the control group was 85.7% and in the experimental group, 92.0%. A high proportion of patients (22.2%) treated with Alprazolam manifested daytime sleepiness, while in the group treated with the G-B standardized extract, daytime sleepiness was found in 4.7%. In conclusion, a G-B standardized extract demonstrated therapeutic effectiveness in patients with GAD, without exhibiting significant difference with Alprazolam, but showing fewer cases of daytime sleepiness. The trial was registered at http://clinicaltrials.gov by identifier: NCT03702803.

Asunto(s)

Trastornos de Ansiedad/tratamiento farmacológico , Galphimia/química , Extractos Vegetales/administración & dosificación , Triterpenos/administración & dosificación , Alprazolam/administración & dosificación , Trastornos de Ansiedad/patología , Método Doble Ciego , Femenino , Humanos , Masculino , Persona de Mediana Edad , Cuestionario de Salud del Paciente/normas , Extractos Vegetales/química , Triterpenos/química

RESUMEN

Galphimia glauca (Cav.) Kuntze is an important endemic plant species, which possesses many medicinal properties and has been used in the Mexican traditional medicine for its sedative, anxiolytic, anticonvulsant, antiasthmatic and antiallergic properties. The therapeutic properties of this plant are mainly due to the presence of diverse bioactive compounds such as flavonoids, triterpenoids, and phenolics. Several triterpenoids and flavonoids compounds have been isolated and identified. Modern studies have demonstrated many biological activities such as anti-inflammatory, antidiarrheal, gastroenteritis, antimalarial and cytotoxic activities. Nevertheless, many studies are restricted to the crude extract, and many bioactive compounds are yet to be identified and validated according to its traditional use. However, its commercial exploitation and use are highly limited due to the non-availability of enough plant material and lack of knowledge about its agronomical practices. Moreover, the misinterpretation and mislabeling of closely related species of the genus Galphimia Cav. as G. glauca or G. gracilis is a common problem for its rigorous scientific study and commercial exploitation. The present review provides comprehensive knowledge based on the available scientific literature. To the best of our knowledge, this is the first review on G. glauca. This comprehensive information will certainly provide a guide for the better understanding and utilization of G. glauca for its scientific and industrial exploitation.

Asunto(s)

Galphimia/química , Extractos Vegetales/química , Extractos Vegetales/farmacología , Plantas Medicinales/química , Animales , Biotecnología , Ensayos Clínicos como Asunto , Evaluación Preclínica de Medicamentos , Ecosistema , Galphimia/clasificación , Humanos , México , Patentes como Asunto , Fitoquímicos/química , Extractos Vegetales/uso terapéutico , Plantas Medicinales/clasificación , Pruebas de Toxicidad

RESUMEN

The aim of this study was to obtain pharmacokinetic data for the anxiolytic compound galphimine-A (G-A) from Galphimia glauca. G-A is the most abundant anxiolytic compound in this plant, while Galphimine-E (G-E) is the most abundant galphimine, but inactive. G-E was transformed chemically into G-A. The pharmacokinetic study was carried out in ICR mice, which were orally administered a single 200 mg/kg dose of G-A. Samples of blood and brain were taken at different times after administration of G-A. Previously, we established the validation of methods for determining the concentration of G-A. The G-A was detected in plasma 5 min after oral administration, and its concentration reached 2.47 µg/mL. Data from concentration-time curves allowed us to establish the main pharmacokinetic parameters in two models: one- and/or two-compartment. C(max) values were 3.33 and 3.42 µg/mL respectively, likewise AUC(0→1440 min) were 1,951.58 and 1,824.95 µg/mL·min. The G-A in brain tissue was noted to cross the blood-brain barrier, reaching C(max) 2.74 µg/mL, T(max) 81.6 min, and then drop gradually to 0.32 µg/mL detected at 24 h. The presence of G-A in brain tissue, confirmed that this anxiolytic compound can access the target organ and acts directly on the CNS.

Asunto(s)

Ansiolíticos/química , Ansiolíticos/farmacocinética , Galphimia/química , Triterpenos/química , Triterpenos/farmacocinética , Administración Oral , Animales , Encéfalo/metabolismo , Cromatografía Líquida de Alta Presión , Estabilidad de Medicamentos , Ratones , Extractos Vegetales/química , Reproducibilidad de los Resultados

RESUMEN

ETHNOPHARMACOLOGY RELEVANCE: Galphimia glauca (Malpighiaceae) is a Mexican plant popularly used as a tranquilizer in the treatment of nervous system disorders, although it is also used to treat other common illnesses. AIM OF THE STUDY: The aim of this investigation is to find out if populations of Galphimia glauca collected in different regions and ecosystems in Mexico actually belong to the same species by using the contemporary technique of DNA barcodes. Our previous metabolic profiling study demonstrates that different collections of this plant obtained from various geographical areas exhibited diverse chemical profiles in terms of the active compounds named Galphimines. We expected the DNA barcodes apart from indicating the different species of Galphimia would indicate the active populations. MATERIALS AND METHODS: We employed matK, rpoC1 and rbcL DNA barcodes to indicate the different species. Furthermore to investigate the possible impact of the several different ecosystems where the seven populations were collected, thin layer chromatography was employed to create a partial chemical profile, which was then compared with the metabolic profiles obtained by (1)H-NMR and multivariate data analysis. RESULTS AND CONCLUSIONS: This study showed that the seven populations here analyzed contain at least three different species of the genus Galphimia, although each individual population is homogeneous. Interestingly our TLC analysis clearly showed that the active populations displayed a distinctively unique chemical profile. This work also showed that the use of DNA barcodes combined with chemical profile analysis is an excellent approach to solve the problems of quality control in the development of Galphimia-based medicines as well as for any breeding programs for this species.

Asunto(s)

ADN de Plantas/genética , Galphimia/genética , Filogenia , Ansiolíticos , Código de Barras del ADN Taxonómico , Hipnóticos y Sedantes , Análisis de los Mínimos Cuadrados , México , Hojas de la Planta , Proteínas de Plantas/genética

RESUMEN

UNLABELLED: Galphimia glauca Cav. has demonstrated anxiolytic activity attributable to nor-seco-triterpenes denominated galphimines, the most active of which is galphimine-B. Galphimine-B inhibits ventral tegmental area dopaminergic neurons and interacts with the serotoninergic system of the dorsal hippocampus. A previous clinical study that administered a G. glauca herbal medicinal product for 4 weeks evidenced high percentages of therapeutic effectiveness and safety in patients with generalized anxiety disorder. Based on the previous findings, the goal of the present study was to evaluate the effectiveness, safety, and tolerability of G. glauca herbal medicinal product administered during 15 weeks in patients with generalized anxiety disorder. STUDY DESIGN: double-blind, randomized, lorazepam-controlled clinical trial. STUDY SUBJECTS: adult males and females, ambulatory, diagnosed with generalized anxiety disorder, with 20 or more points on the Hamilton anxiety scale, without data of depression, and without anxiolytic treatment in the previous month. Interventions were as follows. Experimental treatment: G. glauca herbal medicinal product in capsules containing the dry extract of G. glauca standardized in 0.175 mg of galphimine-B, one or two capsules twice a day, during 12 weeks plus 3 withdrawal weeks, and control treatment: lorazepam 0.5 mg with the same presentation and posology. PRIMARY OUTCOME: anxiolytic effectiveness (≥ 50 % reduction of initial Hamilton anxiety scale score). SECONDARY OUTCOMES: tolerability and safety. One hundred ninety-one patients initiated the study with 94 in the experimental group. One hundred four patients concluded the study, 51 of these in the experimental group. Anxiolytic effectiveness, measured as 0 in a negative case and as 1 in a positive case, was assessed 593 times in the experimental group and 631 in the control; the mean effectiveness observed was 0.686 ± 0.019 vs. 0.588 ± 0.019 (repeated-measures ANOVA; p = 0.0003). In the same way, G. glauca-herbal medicinal product diminished the score in the Hamilton anxiety scale to 11.51 ± 8.27 points and lorazepam to 12.40 ± 8.07 points (repeated-measures ANOVA; p = 0.05). The tolerability analysis, which comprised patients who concluded the treatment plus 11 patients who withdrew due to adverse reactions did not show differences between treatments (p = 0.35), nor did therapeutic safety demonstrate differences between groups (p = 0.21). There were no cases of tolerance, intoxication, dependence, or suppression syndrome. We concluded that G. glauca herbal medicinal product, standardized in 0.175 mg of galphimine-B and administered for 15 weeks to patients with generalized anxiety disorder, showed greater anxiolytic effectiveness than that obtained with lorazepam, with high percentages of therapeutic tolerability and safety.

Asunto(s)

Ansiolíticos/uso terapéutico , Trastornos de Ansiedad/tratamiento farmacológico , Galphimia/química , Lorazepam/uso terapéutico , Extractos Vegetales/uso terapéutico , Triterpenos/uso terapéutico , Adulto , Ansiolíticos/efectos adversos , Método Doble Ciego , Femenino , Humanos , Riñón/efectos de los fármacos , Riñón/metabolismo , Hígado/efectos de los fármacos , Hígado/metabolismo , Lorazepam/efectos adversos , Masculino , Persona de Mediana Edad , Extractos Vegetales/efectos adversos , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Hojas de la Planta/química , Tallos de la Planta/química , Plantas Medicinales/química , Triterpenos/efectos adversos , Triterpenos/química , Triterpenos/aislamiento & purificación

RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: Galphimia glauca has a long traditional use, and continues to be used in Mexico as a natural tranquilizer for the treatment of Central Nervous System disorders as well as for other illnesses. AIM OF THE STUDY: In 2005 the initial use of metabolic profiling to populations of Galphimia glauca resulted in two of the six collected populations being producers for galphimines, the markers for sedative and anxiolytic activities. The aim of this investigation was to confirm the previously established metabolic profile, as well as the previous in vivo results on mice. Additionally in this study we wanted to investigate potential anti-inflammatory properties. MATERIALS AND METHODS: Four years later, we collected samples in the five localities designated for the first-stage investigation in 2005, and in two new locations. Metabolic profiling was carried out by means of (1)H NMR spectroscopy and multivariate data analysis applied to crude extracts from wild plant specimens. HPLC analysis was performed to confirm and quantify the presence of galphimines. Two neuropharmacological in vivo assays on mice were employed to study anxiolytic (elevated plus maze test) and sedative (sodium pentobarbital-induced hypnosis model) activities in the extracts. Anti-inflammatory activity was determined by using the tetradecanoylphorbol acetate-induced mouse ear inflammation model (TPA). RESULTS AND CONCLUSIONS: The results for the 2009 collected species were similar to the 2005 collection, confirming the metabolic profiles and that galphimines are consistent good markers for CNS activity. Galloylquinic acid levels varied between the years without, as of yet, known effects. In vivo anti-inflammatory activity was similar for all plants and thus not linked with galphimines, requiring further studies to identify the active compound(s). Areas of collection affect neuropharmacological activities but not anti-inflammatory action.

Asunto(s)

Ansiolíticos/uso terapéutico , Antiinflamatorios/uso terapéutico , Galphimia , Hipnóticos y Sedantes/uso terapéutico , Extractos Vegetales/uso terapéutico , Animales , Ansiedad/tratamiento farmacológico , Conducta Animal/efectos de los fármacos , Cromatografía Líquida de Alta Presión , Inflamación/tratamiento farmacológico , Espectroscopía de Resonancia Magnética , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos ICR , Fitoterapia , Hojas de la Planta , Análisis de Componente Principal

RESUMEN

Galphimia glauca is popularly employed in Mexico for the treatment of central nervous system disorders. Pharmacological and phytochemical studies have resulted in the identification of the anxiolytic and sedative principle consisting of a mixture of nor-secofriedelanes, named the galphimine series (1 - 9). These active constituents were found in plants collected in the vicinity of a restricted region in Central Mexico, where this species is abundant. A metabolic profiling carried out by means of 1H-NMR spectroscopy and multivariate data analysis was applied to crude extracts from wild plant populations, collected from six different locations as a quality control assessment, in order to differentiate their chemical profile. Principal component analysis (PCA) of the 1H-NMR spectra revealed clear variations among the populations, with two populations out of the six studied manifesting differences, when the principal components PC-1 and PC-2 were analyzed. These two PCs permitted the differentiation of the various sample populations, depending on the presence of galphimines. This information consistently correlated with the corresponding HPLC analysis. The neuropharmacological effects of the crude extracts were evaluated by using ICR mice in the elevated plus maze, as well as the sodium pentobarbital-induced hypnosis models. Both assays demonstrated anxiolytic and sedative responses only among those sample populations which had previously been differentiated by PC-1. Partial least square regression-discriminant analysis (PLS-DA) also confirmed a strong correlation between the observed effects and the metabolic profiles of the plants. The overall results of this study confirm the benefits of using metabolic profiling for the in silico analysis of active principles in medicinal plants.

Asunto(s)

Ansiolíticos/química , Galphimia/química , Hipnóticos y Sedantes/química , Extractos Vegetales/química , Plantas Medicinales/química , Galphimia/metabolismo , Espectroscopía de Resonancia Magnética , México , Análisis Multivariante , Plantas Medicinales/metabolismo

RESUMEN

Galphimia glauca Cav. is a plant used in Mexican traditional medicine as a "nerve tranquilizer". Previous studies have demonstrated that the methanolic extract from this plant species possess an anxiolytic effect. Galphimine B (GB, a nor-seco-triterpene), is the active principle, with an innovative action mechanism. Against this background, a standardized herbal medicinal product was developed from the aqueous extract of G. glauca (GgHP). The present work compared the therapeutic effectiveness, safety, and tolerability of the new GgHP with lorazepam on patients with generalized anxiety disorder (GAD). By means of a controlled, randomized, double-blind clinical trial, outpatients of either sex who matched the DSM-IV diagnostic criteria with a score of > or = 19 points on the Hamilton Anxiety Scale (HAM-A) were included. The experimental group was treated orally with GgHP in capsules twice a day for 4 weeks. The control group received lorazepam (1 mg) under the same conditions and presentation. A total of 152 patients were included in the trial (72 in the experimental group). From the first week of treatment, GgHP showed important anxiolytic effectiveness, very similar to that produced with lorazepam. Both treatments showed therapeutic safety (no alterations on biochemical analysis of hepatic and renal function). Nevertheless, concerning side effects, GgHP evidenced a considerably higher tolerability than lorazepam.

Asunto(s)

Ansiolíticos/farmacología , Trastornos de Ansiedad/tratamiento farmacológico , Galphimia , Fitoterapia , Extractos Vegetales/farmacología , Administración Oral , Adolescente , Adulto , Anciano , Ansiolíticos/administración & dosificación , Ansiolíticos/uso terapéutico , Trastornos de Ansiedad/patología , Método Doble Ciego , Femenino , Humanos , Lorazepam/administración & dosificación , Lorazepam/farmacología , Lorazepam/uso terapéutico , Masculino , Persona de Mediana Edad , Extractos Vegetales/administración & dosificación , Extractos Vegetales/uso terapéutico , Resultado del Tratamiento

RESUMEN

The production of three triterpenoids from Galphimia glauca hairy root cultures, the sedative principle galphimine E (2), the recently described glaucacetalin A (3), and maslinic acid (6), was quantified by HPLC in the biomass and the culture medium. Batch cultures of the hairy root line VYT, obtained through infecting cotyledons with Agrobacterium rhizogenes ATCC 15 834, were grown for 41 days in shake flasks containing B5 medium without phytohormones. A maximum biomass of 11 g/L DW was obtained on day 33, while the doubling time was 6 days. Throughout the growth cycle fresh and dry weights as well as triterpene production were registered. Glaucacetalin A (3), excreted into the culture media, reached a maximum amount of 2.14 mg/L after 21 days while galphimine E (2) and maslinic acid (6) were recovered from the root biomasses reaching maximum concentrations of 0.11 and 0.43 mg/g, respectively, on day 39.

Asunto(s)

Galphimia/metabolismo , Triterpenos/metabolismo , Biomasa , Cromatografía Líquida de Alta Presión , Medios de Cultivo/química , Técnicas de Cultivo , Estructura Molecular , Triterpenos/química , Triterpenos/aislamiento & purificación

RESUMEN

The anxiolytic effects of galphimine B (1), galphimine A (2), and galphimine E (3), natural nor-secofriedelanes isolated from Galphimia glauca, as well as derivatives obtained by acetylation (4), hydrogenation of the C-1/C-2 double bond (5), basic hydrolysis followed by hydrogenation of the C-1/C-2 double bond (6), and deacetylation (7) of galphimine E (3), were evaluated on ICR mice exposed to the elevated plus-maze test. This study also included the evaluation of a galphimines-rich fraction (GRF) with a known concentration of 1-3, obtained from the dry leaves of G. glauca. Intraperitoneal administration of 15 mg/kg of 1, 2, 6, and GRF (1 h before testing) caused an anxiolytic-like effect in the animals, increasing significantly (p <0.001) the percentage of time of permanence and the number of crossings toward the open arms of the plus-maze. No activity was detected after administration of compounds 3, 4, 5, and 7. These results showed that GRF had activity similar to the most active pure galphimines (1 and 2) and that, like for the spasmolytic activity previously reported, the main determining factor responsible for the anxiolytic activity of the compounds was the presence of free hydroxyl groups at C-4, C-6, and C-7 and the presence of the double bond in the A ring.

Asunto(s)

Ansiolíticos/aislamiento & purificación , Ansiolíticos/farmacología , Galphimia/química , Plantas Medicinales/química , Triterpenos/aislamiento & purificación , Triterpenos/farmacología , Animales , Ansiolíticos/química , Modelos Animales de Enfermedad , México , Ratones , Estructura Molecular , Relación Estructura-Actividad , Triterpenos/administración & dosificación , Triterpenos/química

RESUMEN

An infusion prepared with aerial parts from Galphimia glauca has been widely used in Mexican traditional medicine as a remedy for nervous excitement. The sedative activity of a methanolic extract from this plant has been demonstrated by neuropharmacological tests. This effect was attributed to the nor-secotriterpene named galphimine B (GB). In the present work, the anxiolytic and antidepressant-like effects of G. glauca methanolic extract (standardized on GB content, 8.3mg/g) were assayed by using the elevated plus-maze, light-dark test and the forced swimming paradigm, on ICR albino mice. This extract, administered orally, three times (24, 18 and 1h before the test), and in different doses (125, 250, 500, 1,000 and 2,000 mg/kg) was able to increase significantly (p<0.05) the number of entries, as well as the time spent in the open arms of the elevated plus-maze, indicating an anxiolytic-like effect. A similar effect was observed in the light-dark paradigm test, the time spent in the light box was increased in treated mice. Nevertheless, this treatment was unable to change any parameter in the forced swimming test. Altogether, these results suggest an anxiolytic-like effect to the methanolic standardized extract of G. glauca on ICR inbred mice.

Asunto(s)

Ansiolíticos/farmacología , Antidepresivos/farmacología , Galphimia/química , Extractos Vegetales/farmacología , Animales , Ansiolíticos/química , Antidepresivos/química , Relación Dosis-Respuesta a Droga , Masculino , Medicina Tradicional , México , Ratones , Ratones Endogámicos ICR , Componentes Aéreos de las Plantas , Extractos Vegetales/química

RESUMEN

An in vitro micropropagation protocol is described for Galphimia glauca Cav. (Malpighighiaceae). Multiple shoots were formed in vitro from axillary bud explants inoculated on MS medium supplemented with indole-3-acetic acid (IAA) and kinetin (KN) combinations. A maximum of 20 shoots was obtained from a single bud in a 60-day culture period. In vitro-grown shoots were successfully rooted and transferred to field conditions (90 % survival). The sedative triterpenoid galphimine-B (1) content of micropropagated plants transferred to field conditions was similar to that of wild plants. Our results suggest that the in vitro propagation protocol described here will have positive effects on conservation of natural resources as well as on adequate techniques for multiplication of an important Mexican medicinal plant.

Asunto(s)

Galphimia/química , Galphimia/crecimiento & desarrollo , Hipnóticos y Sedantes/análisis , Triterpenos/análisis , Hipnóticos y Sedantes/química , Hipnóticos y Sedantes/aislamiento & purificación , Ácidos Indolacéticos/farmacología , Cinetina/farmacología , Estructura Molecular , Farmacognosia/métodos , Extractos Vegetales/química , Triterpenos/química , Triterpenos/aislamiento & purificación

RESUMEN

Preparative-scale recycling HPLC was used for the complete resolution of a complex mixture of nor-secofriedelanes into five major peaks (I-V) from the sedative methanolic extracts prepared from the aerial parts of Galphimia glauca. Argentation chromatography was used to show peaks I, II, IV, and V to be mixtures of isomers around the E-ring double bond, represented by the endocyclic C-20, C-21 double-bond isomers, galphimines A (3), B (1), D (4), and E (2), and the C-20, C-29 exocyclic forms, galphimines F-I (5-8). Galphimine C (9), isolated from peak III, corresponded to the C-19, C-20 double-bond isomer of the previously known major sedative constituent galphimine B. The characterization of all the new triterpenes (3-9) was performed primarily by high-field NMR spectroscopy. Comparison between experimental and calculated (1)H-(1)H vicinal coupling constants and the analysis of molecular mechanics structures revealed that the ring B of these compounds exists in a boatlike conformation. The absolute configuration for the stereogenic carbinol center at C-4 was established by the application of the Mosher ester derivatization technique carried out in NMR tubes.

Asunto(s)

Galphimia/química , Hipnóticos y Sedantes/aislamiento & purificación , Plantas Medicinales/química , Triterpenos/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Hipnóticos y Sedantes/química , Hipnóticos y Sedantes/farmacología , México , Modelos Moleculares , Conformación Molecular , Estructura Molecular , Resonancia Magnética Nuclear Biomolecular , Triterpenos/química , Triterpenos/farmacología

RESUMEN

Transformed root cultures of Galphimia glauca (Malpighiaceae) were established by infecting cotyledons and hypocotyls with Agrobacterium rhizogenes ATCC 15 834. Cotyledon-derived cell lines were grown in liquid B5 nutrient medium without phytohormones and have shown the typical hairy roots phenotype over two years of continuous subculturing. PCR analysis was used to confirm the integration of rol A and rol C genes into the plant genome. The transformed cultures synthesized three major norfriedelanes, the new glaucacetalins A-C (1-3), which were secreted into the nutrient medium. The structural elucidation of these in vitro produced metabolites was performed by the application of high resolution NMR techniques that proved them to be triterpenoids related to the known galphimines, the sedative principles of this plant species. These results suggest the possibility of further biotechnological exploration of sedative friedelane biosynthesis by in vitro plant organ cultures.

Asunto(s)

Galphimia/genética , Fitoterapia , Plantas Medicinales/genética , Rhizobium , Triterpenos/metabolismo , Técnicas de Cultivo de Célula , Galphimia/metabolismo , Humanos , Raíces de Plantas , Plantas Modificadas Genéticamente , Plantas Medicinales/metabolismo

Asunto(s)

Galphimia , Rinitis Alérgica Estacional

Asunto(s)

Humanos , Animales , Cobayas , Técnicas In Vitro , Dosis Mínimas , Investigación Homeopática Básica , Mecanismo de Acción del Medicamento Homeopático , Calorimetría , Técnicas de Cultivo , Intoxicación por Cadmio , Cisplatino/envenenamiento , Anuros , Tiroxina/farmacología , /farmacología , Factor de Crecimiento Epidérmico/farmacología , Fibroblastos , Cólera/terapia , Medicina de Hierbas , Galphimia/uso terapéutico , Rinitis Alérgica Estacional/terapia , Ensayos Clínicos como Asunto , Triticum , Argentum Nitricum/farmacología , Conformación Molecular , Adyuvantes Inmunológicos , Carcinógenos , Neoplasias/terapia