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Chemical profiling of Buddleja globosa was performed by high-performance liquid chromatography coupled to electrospray ionization (HPLC-DAD-ESI-IT/MS) and quadrupole time-of-flight high-resolution mass spectrometry (HPLC-ESI-QTOF/MS). The identification of 17 main phenolic compounds in B. globosa leaf extracts was achieved. Along with caffeoyl glucoside isomers, caffeoylshikimic acid and several verbascoside derivatives (β-hydroxyverbascoside and β-hydroxyisoverbascoside) were identified. Among flavonoid compounds, the presence of 6-hydroxyluteolin-7-O-glucoside, quercetin-3-O-glucoside, luteolin 7-O-glucoside, apigenin 7-O-glucoside was confirmed. Campneoside I, forsythoside B, lipedoside A and forsythoside A were identified along with verbascoside, isoverbascoside, eukovoside and martynoside. The isolation of two bioactive phenolic compounds verbascoside and forsythoside B from Buddleja globosa (Buddlejaceae) was successfully achieved by centrifugal partition chromatography (CPC). Both compounds were obtained in one-step using optimized CPC methodology with the two-phase solvent system comprising ethyl acetate-n-butanol-ethanol-water (0.25: 0.75: 0.1: 1, v/v). Additionally, eight Natural Deep Eutectic Solvents (NADESs) were tested for the extraction of polyphenols and compared with 80% methanol. The contents of verbascoside and luteolin 7-O-glucoside after extraction with 80% methanol were 26.165 and 3.206 mg/g, respectively. Among the NADESs tested in this study, proline- citric acid (1:1) and choline chloride-1, 2- propanediol (1:2) were the most promising solvents. With these NADES, extraction yields for verbascoside and luteolin 7-O-glucoside were 51.045 and 4.387 mg/g, respectively. Taken together, the results of this study confirm that CPC enabled the fast isolation of bioactive polyphenols from B. globosa. NADESs displayed higher extraction efficiency of phenolic and therefore could be used as an ecofriendly alternative to classic organic solvents.

Buddleja globosa Hope (Buddlejaceae) is a native species cultivated in Chile, Peru and Argentina. In folk medicine, this plant is frequently used for the treatment of different wounds (internal and external), as well as intestinal and liver diseases. There is growing scientific evidence on the growing conditions for Buddleja globosa and how its cultivation could be optimized. Methods for its vegetative propagation have also been successfully elaborated. The leaves contain phenylpropanoids, iridoids, terpenes and flavonoids. Ethnopharmacological validation studies suggest that phenylpropanoids and flavonoids are the main and most important compounds in defining the therapeutic activity of Buddleja globosa. This chemical composition explains its pharmacological activity as antioxidant, anti-inflammatory, antimicrobial and, in particular, its wound healing effects. Due to its undeniable potential as a domesticated, cultivated medicinal plant and its remarkable chemical composition, it is still necessary to develop its properly standardized, clinically validated phytopharmaceutical products.

Over the last few years, the green synthesis of nanoparticles (NPs) using plant extracts has emerged as a promising methodology for the fabrication of metallic NPs (especially silver, copper, and gold NPs), as it involves an easy, fast, low-cost, and environmentally friendly bioprocess. However, many factors affect the sizes and morphologies of NPs biosynthesized by this method, including the nature of the plant extract, among others. Therefore, the green synthesis of metal NPs with defined stability, size, and morphology distribution remains under evaluation. In the present study, we propose aqueous extracts from the endemic-medicinal plant Budleja globosa (“Matico”) as an efficient bioproduct for the green synthesis of silver NPs (AgNPs). Experimental results indicate that room temperature, low concentrations of leaf extracts of B. globosa, and silver nitrate salt were sufficient to biosynthesize AgNPs with uniform size (16 nm) and shape distribution (spherical).

Six native plants of South America traditionally consumed in the Patagonian region (southern Argentina and Chile), namely: Adesmia boronioides Hook. f., Apium australe Thouars, Buddleja globosa Hope, Drimys andina (Reiche) R. Rodr. & Quezada, Dysphania multifida L. and Solidago chilensis Meyen were investigated to determine the nutraceutical properties of infusions of their aerial parts. The infusions were characterized in terms of their antioxidant activity, phenolic and flavonoid content, profile of phenolic compounds, general toxicity and cytotoxicity on two different human cell lines: T84 (derived from colon cancer) and HTR8/SVneo (not derived from cancer). Twenty-nine compounds, mainly phenolic acids and flavonoids, were identified. This is the first analysis of phenolic compounds in infusions from native plants of Patagonia. D. andina, B. globosa and S. chilensis showed high levels of antioxidants, even higher than those of Green Tea. The content of phenolic compounds correlated significantly with the antioxidant activity of the samples analyzed. The toxicity test indicated that the use of A. australe, B. globosa and D. multifida seems safe, but a moderate consumption is suggested for A. boronioides, D. andina and S. chilensis until more exhaustive and long-term results are available. Moreover, A. boronioides and S. chilensis showed anticancer potential due to their antiproliferative activity on human cancer cell lines. Fil: Gastaldi, Bruno. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de la Patagonia "San Juan Bosco". Facultad de Ciencias Naturales - Sede Esquel; Argentina Fil: Marino, Gabriela Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina Fil: Assef, Yanina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Centro de Investigación Esquel de Montaña y Estepa Patagóica. Universidad Nacional de la Patagonia "San Juan Bosco". Facultad de Ciencias Naturales - Sede Esquel. Centro de Investigación Esquel de Montaña y Estepa Patagónica; Argentina Fil: Silva Sofrás, Fresia Melina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de la Patagonia "San Juan Bosco"; Argentina Fil: Catalan, Cesar Atilio Nazareno. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Química del Noroeste. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química del Noroeste; Argentina Fil: González, Silvia Beatriz. Universidad Nacional de la Patagonia "San Juan Bosco". Facultad de Ciencias Naturales - Sede Esquel; Argentina

Ethnopharmacological relevance Leaf extracts of Buddleja globosa (Buddlejaceae) are used in Chilean folk medicine for wound healing. The anti-inflammatory (topic and per os), analgesic (per os) effects and the antioxidant activity of Buddleja globosa were for the first time reported by us. Aim of the study Assess the antinociceptive activity of the methanol sequential and global extracts using complementary chemical and thermal models of pain, characterize pharmacologically the antinociception induced, evaluate seasonal influence to support Buddleja globosa medicinal use. Materials and methods Global methanol, sequential methanol and ethanol (leaves collected in autumn and summer) extracts were evaluated for oral and topic analgesia in tail flick, formalin and writhing models, verbascoside and 7-O-luteolin glucoside were assayed in tail flick and writhing. Ibuprofen was used as reference. For characterization of induced antinociception, naltrexone, naltrindole, tropisetron, nor-binaltorphimine, prazosin, yohimbine, atropine, and N-nitro- l -arginine methyl ester were used as antagonists and inhibitors drugs. Results Seasonal influence was observed since autumn extract resulted less active. Extracts showed a dose-dependent antinociceptive activity in all assays, the highest effects were obtained for the formalin and writhing test. Verbascoside was more active than ibuprofen in the writhing test (67.6% and 50.0% at equimolar doses) and showed similar effects in the tail flick (topic and oral) near 25% at equivalent doses – ED25 or EC25 – to ibuprofen. Luteolin 7-O-glucoside was slightly more active in the tail flick test and nearly half active than verbascoside in the writhing assay. Effectiveness was higher for the sequential than for global alcoholic extracts, and can be increased by selective blocking of opioid receptors. Global methanol extract seems modulated only by naltrexone. Conclusions Analgesic effect of Buddleja globosa is here demonstrated validating its use in traditional medicine. Season influence is important to be considered.

In vitro tests are now widely employed in ethnopharmacological research because of ethical reasons and their usefulness in bioactive-guided fractionation and determination of active compounds. For many disease conditions, a variety of in vitro tests can now be employed as the biochemical mechanisms underlying disease and healing processes are understood. Approaches to the in vitro investigations of wound healing processes are exemplified by studies on extracts of Buddleja species and three Ghanaian species Spathodea campanulata, Commelina diffusa and Secamone afzelii. Most studies have been carried out on Buddleja officinalis or Buddleja globosa. The extracts have been shown to have anti-inflammatory and antioxidant properties due to flavonoids, triterpenoids, diterpenoids and caffeic acid derivatives. There appears to a slight effect on proliferation of fibroblasts at lower concentrations, but this was not significant, and higher concentrations appeared to be cytotoxic. Novel findings are the ability of Buddleja globosa leaf extracts to induce differentiation in keratinocytes and to alter the profile of proteins produced by cultured fibroblasts. Extracts also had some effect on lattice contraction. The three Ghanaian species examined show a mixture of antimicrobial and antioxidant activities. The evolution over recent years of tests for wound healing, from in vivo tests to cell-based systems and chemical reactions and on to investigations into effects on secondary messengers and protein expression, is described.

An aqueous extract of Buddleja globosa leaves, used traditionally in Chile for wound healing, was tested for the ability to stimulate growth of fibroblasts in vitro and for antioxidant activity in the same fibroblast cell system challenged with hydrogen peroxide. Low concentrations of the extract gave an increase in fibroblast growth which was not statistically significant but cytotoxicity was observed at concentrations greater than 50 microg/ml. The extract showed strong antioxidant effect and fractionation led to the isolation of three flavonoids and two caffeic acid derivatives, each of which was shown to contribute to the antioxidant effect at concentrations below 10 microg/ml. These activities would accelerate the healing of wounds.

The aim was to produce a tetraploid form of Buddleia globosa to facilitate introgression of yellow flower colour into B. davidii, which is naturally tetraploid. Protocols were established for the micropropagation of B. globosa and tetraploid plants were obtained by application in vitro of colchicine to pre-cultured excised nodal sections. Three concentrations of colchicine were applied (0.01%, 0.05% and 0.1% w/v) for 1, 2 or 3 days. At 0.01% tetraploids were produced only after 2 days of application. All other treatments produced at least one tetraploid. The colchicine technique was extremely effective: of 29 lines tested, 19 were tetraploid and 5 were mixoploid. The vegetative characteristics of these tetraploids are described and the flowering characteristics of the three that flowered.

Ethnopharmacological relevance Buddleja globosa , known as “matico”, is employed in Chile for wound healing. Aim of the study To validate the traditional use of the crude drug through in vivo and in vitro evaluation of the anti-inflammatory, analgesic and antioxidant properties of its extracts. Materials and methods Sequential hexane, dichloromethane, methanol and total methanol extracts were studied using bioguided fractionation. The following activities were investigated: analgesic (writhing test), oral and topic anti-inflammatory (paw- and ear-induced edema), free radical scavenging and antioxidant activities (1,1-diphenyl-2-picrylhydrazyl, DPPH, superoxide anion, lipid peroxidation and xanthine oxidase inhibition). Sodium naproxen, nimesulide, indomethacin were used as reference drugs for in vivo , quercetin and allopurinol for in vitro assays. Results A mixture of α- and β-amyrins was isolated from the hexane extract that showed 41.2% of analgesic effect at 600 mg/kg, inhibited by 47.7 and 79.0% the arachidonic acid (AA) and 12-deoxyphorbol-13-decanoate (TPA)-induced inflammation at 3 mg/20 μL/ear, respectively. A mixture of β-sitosterol, stigmasterol, stigmastenol, stigmastanol and campesterol was isolated from the fraction CD4-N and β-sitosterol-glycoside from the fraction CD5-N, reducing TPA-induced inflammation by 78.2 and 83.7% at 1 mg/20 μL/ear, respectively. The fraction CD4-N at 300 mg/kg also showed analgesic activity (38.7%). The methanol extract at 600 mg/kg per os showed anti-inflammatory effect (61.4%), topic anti-inflammatory (56.7% on TPA) and analgesic activity (38.5%). Verbascoside and luteolin-7- O -glucoside were the major components of the methanol extract; apigenin 7- O -glucoside was also detected. Inhibition of superoxide anion, lipoperoxidation, and DPPH bleaching effect was found in the methanol serial and global extracts. Conclusions The present report demonstrate the analgesic and anti-inflammatory properties of Buddleja globosa and validate its use in Chilean traditional medicine.

Numerous investigations exist about the alterations that oxygen free radicals can provoke on biomolecules; these modifications can be prevented and/or reversed by different antioxidants agents. On the other hand, 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), a stable nitrogen synthetic radical, is used to evaluate the antioxidant capacity of medicinal herbal products; however, the structural changes that this radical provoke on the herbal active principles are not clear yet. In this work, we compared the redox reactivity of oxygen free radicals and DPPH radical on phospholipids and protein thiol groups present in rat liver microsomes. Cu2+/ascorbate was used as generator system of oxygen free radical and as antioxidant, an extract of Buddleja globosa's leaves. Cu2+/ascorbate provoked microsomal lipid peroxidation, microsomal thiols oxidation and oxygen consumption; all of these phenomena were inhibited by B. globosa extract. On the other hand, DPPH was bleached in different extension by the herbal extract and phosphatidyl choline; beside, DPPH decreased microsomal thiols content, but this phenomenon were not prevented by the herbal extract. Furthermore, DPPH did not induce oxygen consumption and neither modified the oxygen consumption induced by Cu2+/ascorbate. Distinct redox mechanisms may explain the differences between the reactivity of DPPH and oxygen free radicals on biomolecules, which is discussed.

The aqueous extracts of three species of Buddleja which are used in traditional medicine for the treatment of liver ailments have shown an inhibitory effect against induced cytotoxicity of cultured hepatocytes. Fractionation of one of these extracts, from B. globosa leaves, resulted in the isolation of five iridoids, two phenylpropide glycosides, and a flavonoid glycoside. Two of the iridoids were found to be novel compounds, 7-p-methoxycinnamoylaucubin (1) and 7-p-methoxycinnamoylcatalposide (2). One of the phenylpropides, echinacoside, has not previously been reported from Buddleja. Testing of the isolated compounds in the cultured hepatocyte system indicated that the activity is most likely due to the flavonoid and phenylpropide constituents. Phenylpropide glycosides have not previously been shown to have a liver-protectant effect.

A chloroform extract of the stem bark of Buddleja globosa yielded a mixture of crystalline compounds which were found to be esters of long chain fatty acids and 4-hydroxycinnamyl alcohol or 2-(4-hydroxyphenyl ethanol. This type of ester has been proposed as forming the units of the complex polymer suberin but no individual monomers have been reported until now.