Estudo químico de Hedyosmum brasiliense Miq., Chloranthaceae

Tese (doutorado) - Universidade Federal de Santa Catarina, Centro de Ciências da Saúde, Programa de Pós-Graduação em Farmácia, Florianópolis, 2015.Hedyosmum brasiliense Miq., Chloranthaceae é um arbusto dioico e endêmico no Brasil. Pertencente à família Chloranthaceae, usa-se popularmente como calmante, anti-inflamatório e afrodisíaco na forma de chá ou macerado em vinho branco. O presente trabalho é uma continuação dos estudos anteriores que teve como objetivo estudar fitoquimicamente as frações diclorometano, acetato de etila e aquosa do extrato etanólico e também o extrato aquoso desta espécie. Cinco sesquiterpenoides (um inédito) e onze fenólicos foram isolados e caracterizados neste trabalho. Das frações diclorometano, acetato de etila e aquosa foram isoladas e caracterizadas seis substâncias que estão sendo descritas pela primeira vez para Hedyosmum brasiliense (aromadendrane-4?,10a-diol, ácido rosmarínico, rosmarinato de etila, canferol, canferol-3-?-O-glucuronídeo e oresbiusina) e três substâncias (3,4-dihidroxibenzaldeído, ácido 4-hidroxi-3-metoxibenzoíco e escopoletina) que já foram descritas previamente para Hedyosmum brasiliense. Entre estas substâncias podoandina, aromadendrano-4?,10a-diol e onoseriolídeo foram avaliadas contra o efeito induzido pelo peptídeo ß-amiloide 1-42 (Aß1-42) na tarefa de esquiva passiva, um modelo de Alzheimer em camundongos. Do extrato aquoso a lactona sesquiterpênica inédita 1-?-acetoxyeudesma-3,7(11)  dien  8,12  olíde foi isolada e caracterizada. Ainda, pelo método desenvolvido por cromatografia de partição centrífuga, ácido rosmarínico foi isolado em uma etapa única. Além disso, ácido isorínico, urolignosídeo, (7S,8R)-5-metoxidiidrodesidrodiconiferil álcool-4-O-ß-D-glicopiranosídeo, canferol, canferol-3-?-O-D-glucuronídeo, podoandina, onoseriolídeo e 15-hidroxi-isogermafurenolídeo foram isolados e identificados. O extrato aquoso e o ácido rosmarínico foram avaliados contra Mycobacterium tuberculosis. As substâncias foram elucidadas através de espectroscopia de ressonância magnética nuclear e espectrometria de massas, em comparação com dados já publicados. A metodologia desenvolvida por cromatografia líquida acoplada à espectrometria de massas com ionização por eletrospray proporcionou a quantificação do ácido rosmarínico (188 mg/g do extrato aquoso), onoseriolídeo (1,7 mg/g do extrato aquoso) e podoandina (1,9 mg/g do extrato). Quanto aos testes biológicos, a podoandina, aromadendrano-4?,10a-diol e onoseriolídeo reverteram significativamente o efeito induzido pelo peptídeo ß-amiloide 1-42 (Aß1-42) na tarefa de esquiva passiva, um modelo de Alzheimer em camundongos. Ainda a podoandina, onoseriolídeo e 15-acetoxi-isogermafurenolídeo relaxaram os anéis aórticos e corpo cavernoso. Entretanto, a avaliação do extrato aquoso e ácido rosmarínico contra Mycobacterium tuberculosis não mostrou nenhuma atividade. Abstract : Hedyosmum brasiliense Miq., Chloranthaceae is a dioecious shrub and endemic in Brazil. Belonging to the Chloranthaceae family, it is popularly used as a sedative, anti-inflammatory and aphrodisiac in the form of tea or macerated in white wine. The present work is the continuation of previous studies that has as its objective to study phytochemically the dichloromethane, ethyl acetate and water fractions of the ethanol extract and the aqueous extract of this species. Five sesquiterpenoids (one new) and eleven phenolic compounds were isolated and characterized in this work. Six substances that were isolated and characterized from the dichloromethane, ethyl acetate and aqueous fractions, are being described for the first time for this species (aromadendrane-4?, 10a-diol, rosmarinic acid, ethyl rosmarinate, kaempferol, kaempferol-3 -O -?-D -glucuronide and oresbiusin) and three substances (3,4-dihydroxybenzaldehyde, 4-hydroxy-3-methoxy benzoic acid and scopoletin) which have been previously described for this species. Among these substances, podoandin aromadendrane-4?, 10a-diol and onoseriolide were evaluated against the ß-amyloid peptide 1-42 (Aß1-42) in the passive avoidance task, a model of Alzheimer in mice. From the aqueous extract the new sesquiterpene lactone 1-?-acetoxyeudesma-3,7 (11) - dien  8,12 - olide was isolated and characterized. Furthermore, through a one-step method by centrifugal partition chromatography rosmarinic acid was isolated from the aqueous extract. Moreover, isorinic acid, urolignoside, (7S, 8R) -5-alcohol methoxydihydrodehydrodiconiferyl-4-O-ß-D-glucopyranoside, kaempferol, kaempferol-3-O-?-D-glucuronide, podoandin, onoseriolide and 15-hydroxy-isogermafurenolide were isolated and identified as already known substances. The aqueous extract and rosmarinic acid were evaluated against Mycobacterium tuberculosis, while podoandina, onoseriolide and 15-acetoxy-isogermafurenolide were evaluated. The compounds were elucidated by nuclear magnetic resonance spectroscopy and mass spectrometry and in comparison with previously reported data. A methodology based on liquid chromatography coupled to mass spectrometry with electrospray ionization, was used for the quantification of rosmarinic acid (188 mg/g of aqueous extract), onoseriolide (1.7 mg/g of aqueous extract) and podoandin (1.9 mg/g of the aqueous extract). For biological tests, podoandin aromadendrane-4?, 10a-diol and onoseriolide significantly ameliorated the damage caused by the ß-amyloid peptide 1-42 (Aß1-42) in the passive avoidance task, a model of Alzheimer in mice. Moreover, podoandin, onoseriolide and 15-acetoxy-isogermafurenolide relaxed aortic rings and corpus cavernosum. However, the evaluation of the aqueous extract and rosmarinic acid against Mycobacterium tuberculosis did not demonstrate any activity

Investigação fitoquímica do extrato etanólico de Hedyosmum brasiliense Miq., Chloranthaceae

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências da Saúde, Programa de Pós-graduação em Farmácia, Florianópolis, 2011Hedyosmum brasiliense é um arbusto aromático que pertence à família Chloranthaceae e é popularmente usado como calmante e antiinflamatório, para tratar frieiras, enxaquecas e doenças de ovário. Este trabalho visou continuar a análise fitoquímica desta espécie que é conhecida popularmente como "Cidrão". Foi possível isolar e identificar treze substâncias oriundas só das frações hexano e diclorometano do extrato etanólico. Dessas substâncias, quatro são inéditas na literatura e pertencem à classe das lactonas sesquiterpênicas 15-acetilisogermafurenolídeo (H622), 1-hidroxi-10,15-metilenopodoandina (HB 212), 15-hidroxi-isogermafurenolídeo (HB 21) e brasiliensolídeo (HL 33) que é uma lactona sesquiterpênica dimérica. Seis estão sendo descritas pela primeira vez para a espécie (monoglicerídeo de ácido esteárico, éster etílico do ácido linolênico, éster etílico do ácido linoléico, ácido 3-metoxi-4-hidroxibenzoico, escopoletina e vanilina), e três (podoandina, 13-hidroxi-8,9-diidroshizukanolídeo (13HDS) e ß- sitosterol) já foram descritas previamente para H. brasiliense. Quanto aos estudos farmacológicos, a podoandina mostrou atividade antidepressiva agindo sobre as vias adrenérgicas, dopaminérgicas e serotonérgicas; o extrato bruto e as lactonas, podoandina e 13HDS exibiram a atividade vasodilatadora e as lactonas sesquiterpênicas (H622, HB 212 e HB 21) não tiveram atividade antimicrobiana contra Mycobacterium tuberculosis. As substâncias foram identificadas pela espectroscopia de ressonância magnética nuclear de 1D, 2D, espectrometria de massas e dados da literatura.Hedyosmum brasiliense is an aromatic shrub that belongs to the Chloranthaceae family and is popularly used as a calmative and ntiinflammatory, for treating athlete's foot, migraines and diseases of the ovary. This work was done to continue the phytochemical analysis of this species which is popularly known as "Cidrão". It was possible to isolate and identify thirteen substances which were obtained from the hexane and dichloromethane fractions of the ethanol extract. Out of these substances, four are new and belong to the class of sesquiterpene lactones: 15-acetyl-isogermafurenolide (H622), 1-hydroxy-10 ,15-methylenepodoandin (HB 212), 15-hydroxy-isogermafurenolide (HB21) and brasiliensolide (HL 33) which is a dimeric sesquiterpene lactone. Six are being described for the first time for this species (stearic acid monoglyceride, linolenic acid ethyl ester, linoleic acid ethyl ester, 3-methoxy-4- hydroxybenzoic acid, scopoletin and vanillin), and three (podoandin, 13-hydroxy-8,9-dehydroshizukanolide (13HDS) and ß-sitosterol) were previously described for H. brasiliense. With regards to pharmacological studies, podoandin exhibited antidepressant activity by acting on adrenergic, dopaminergic and serotonergic pathways; the crude extract and the lactones podoandin and 13HDS exhibited vasodilator activity and the sesquiterpene lactones (H622, HB 212 and HB 21) showed no antimicrobial activity against Mycobacterium tuberculosis. The substances were identified by Nuclear magnetic resonance spectroscopy 1D, 2D, mass spectrometry and already published data

Effects of the topical application of an ethyl acetate fraction from Vernonia scorpioides on excisional wounds infected with Staphylococcus aureus in rats

We investigated the healing process on excisional wounds infected with Staphylococcus aureus in rats, treated with 50 µL of ethyl acetate III from Vernonia scorpioides (Lam.) Pers., Asteraceae, rifamycin diethylamide B 25 mg, or saline. The lesions were measured daily and after seven days were surgically removed and histologically processed. The results indicate a favorable action of the EAIII, demonstrated by the increased wound contraction, smaller area of necrotic tissue, good development of granulation tissue, extensive extracellular matrix deposition and epithelial regeneration. This sub-fraction was phytochemically investigated in parallel studies, revealing the presence of sesquiterpene lactones (glaucolides and hirsutinolides) such as diacethylpiptocarphol and related hirsutinolides, flavonoids and cinnamic acid derivatives and also a new polyacetylene, which have been previously published. Results support the effectiveness of V. scorpioides antimicrobial activity in infected wound healing in rats

Chitosan-Based Biomaterial, Calcium Hydroxide and Chlorhexidine for Potential Use as Intracanal Medication

The objective of this study was to develop a chitosan-based biomaterial with calcium hydroxide and 2% chlorhexidine for intracanal treatment application and, consequently, to diminish the number of microorganisms in the root canal system. The chitosan solution was prepared by dissolving it in 2% and 4% acetic acid (v/v) for 1 h at room temperature (25 °C) with magnetic agitation (430 rpm). Calcium hydroxide was obtained in two stages: the first was the synthesis of the calcium oxide—CaO, and the second was that of the calcium hydroxide—Ca(OH)2. The samples were developed using different concentrations of chitosan, calcium hydroxide, and chlorhexidine 2%. They were codified as Ca(OH)2 + Q2% (M1), Ca(OH)2 + Q4% (M2), Ca(OH)2 + Q2% + CLX (M3), Ca(OH)2 + Q4% + CLX (M4), Ca(OH)2 + Q2% + PEG (M5), and Ca(OH)2 + Q4% + PEG (M6). They were characterized through Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and rheological measurement, and the antimicrobial activity was evaluated in vitro. Characteristic absorption bands of the source materials used in this research were observed in the FTIR spectra. The X-ray diffraction technique indicated that the material has a semi-crystalline structure and that the presence of calcium hydroxide made the biomaterial more crystalline. The viscosity measurement showed a pseudoplastic behavior of the studied samples. The microbiologic analysis was positive for all samples tested, with bigger inhibition zones for the samples M3 and M4. As a result, we conclude that the formulation developed based on chitosan is promising and has potential to be an intracanal medication

Chitosan-Based Biomaterial, Calcium Hydroxide and Chlorhexidine for Potential Use as Intracanal Medication

The objective of this study was to develop a chitosan-based biomaterial with calcium hydroxide and 2% chlorhexidine for intracanal treatment application and, consequently, to diminish the number of microorganisms in the root canal system. The chitosan solution was prepared by dissolving it in 2% and 4% acetic acid (v/v) for 1 h at room temperature (25 °C) with magnetic agitation (430 rpm). Calcium hydroxide was obtained in two stages: the first was the synthesis of the calcium oxide—CaO, and the second was that of the calcium hydroxide—Ca(OH)2. The samples were developed using different concentrations of chitosan, calcium hydroxide, and chlorhexidine 2%. They were codified as Ca(OH)2 + Q2% (M1), Ca(OH)2 + Q4% (M2), Ca(OH)2 + Q2% + CLX (M3), Ca(OH)2 + Q4% + CLX (M4), Ca(OH)2 + Q2% + PEG (M5), and Ca(OH)2 + Q4% + PEG (M6). They were characterized through Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and rheological measurement, and the antimicrobial activity was evaluated in vitro. Characteristic absorption bands of the source materials used in this research were observed in the FTIR spectra. The X-ray diffraction technique indicated that the material has a semi-crystalline structure and that the presence of calcium hydroxide made the biomaterial more crystalline. The viscosity measurement showed a pseudoplastic behavior of the studied samples. The microbiologic analysis was positive for all samples tested, with bigger inhibition zones for the samples M3 and M4. As a result, we conclude that the formulation developed based on chitosan is promising and has potential to be an intracanal medication

A novel N95 respirator with chitosan nanoparticles: mechanical, antiviral, microbiological and cytotoxicity evaluations

Abstract Background It is known that some sectors of hospitals have high bacteria and virus loads that can remain as aerosols in the air and represent a significant health threat for patients and mainly professionals that work in the place daily. Therefore, the need for a respirator able to improve the filtration barrier of N95 masks and even inactivating airborne virus and bacteria becomes apparent. Such a fact motivated the creation of a new N95 respirator which employs chitosan nanoparticles on its intermediate layer (SN95 + CNP). Results The average chitosan nanoparticle size obtained was 165.20 ± 35.00 nm, with a polydispersity index of 0.36 ± 0.03 and a zeta potential of 47.50 ± 1.70 mV. Mechanical tests demonstrate that the SN95 + CNP respirator is more resistant and meets the safety requisites of aerosol penetration, resistance to breath and flammability, presenting higher potential to filtrate microbial and viral particles when compared to conventional SN95 respirators. Furthermore, biological in vitro tests on bacteria, fungi and mammalian cell lines (HaCat, Vero E6 and CCL-81) corroborate the hypothesis that our SN95 + CNP respirator presents strong antimicrobial activity and is safe for human use. There was a reduction of 96.83% of the alphacoronavirus virus and 99% of H1N1 virus and MHV-3 betacoronavirus after 120 min of contact compared to the conventional respirator (SN95), demonstrating that SN95 + CNP have a relevant potential as personal protection equipment. Conclusions Due to chitosan nanotechnology, our novel N95 respirator presents improved mechanical, antimicrobial and antiviral characteristics