Antinociceptivna i protuupalna svojstva vodeno-etanolnog ekstrakta pokožice grožđa vrste Vitis labrusca izolirane iz otpada vinske industrije

Research background. Extracts from grape pomace, including the wine, show many biological effects such as antioxidant and anti-inflammatory activities. Unfortunately, winemakers discard the bagasse, so the waste is not exploited, although it contains bioactive compounds with antioxidant and anti-inflammatory properties. The work aims to analyze the hydroethanolic extract of peels from Vitis labrusca agro-industrial waste and to evaluate its antinociceptive and anti-inflammatory properties. This study is relevant for reusing a residue and adding value to the grape economic chain. Experimental approach. A representative sample of pomace was obtained and the peels were used to produce the extract. The phenolic compounds were determined by mass spectrometry in multiple reaction monitoring mode and Folin-Ciocalteu colorimetric method, using gallic acid as standard. The biological analyses were carried out using mice orally treated with crude extract at doses of 30, 100 and 300 mg/kg. We evaluated mechanical hyperalgesia by the von Frey method, thermal heat hyperalgesia using a hot plate at 55 °C, paw edema using a pachymeter, and neutrophil recruitment by measurement of myeloperoxidase activity. The nephrotoxicity and hepatotoxicity were evaluated by biochemical analyses using blood samples that were collected after the Vitis labrusca administration. Results and conclusions. In all wet winemaking residues peel mass fraction was 75%, and in dry residues 59%. We identified nine anthocyanins (3-O-glucosides: peonidin, delphinidin, petunidin and malvidin; 3-p-coumaroyl-glucosides: cyanidin, peonidin, petunidin and malvidin, and malvidin-3,5-diglucoside), five flavonoids (apigenin-7-glucoside, luteolin-7-glucoside, quercetin-3-galactoside, isorhamnetin-3-glucoside and myricetin-3-rutinoside), and mass fraction of phenolic compounds, expressed as gallic acid equivalents, was 26.62 mg/g. In vivo assays showed that Vitis labrusca extract at mass fractions 100 and 300 mg/kg reduced carrageenan-induced mechanical and thermal hyperalgesia, 50% of the paw edema, and neutrophil recruitment. In addition, there were no indications of nephrotoxicity and hepatotoxicity. Our extract obtained from winemaking residue has analgesic and anti-inflammatory properties, related at least in part to the presence of phenolic compounds, and it is not toxic to renal and hepatic tissues. Novelty and scientific contribution. This bio-product can be used as an alternative to synthetic anti-inflammatory agents with the same pharmacological potential and fewer side effects. We demonstrated that Vitis labrusca winemaking waste can be used for the production of antinociceptive and anti-inflammatory products (nutraceutical, pharmaceutical and cosmetics) without toxicity, contributing to the environmental economy.Pozadina istraživanja. Ekstrakt komine grožđa, kao i vino, ima mnoga biološka svojstva, poput antioksidacijskog i protuupalnog učinka. Nažalost, proizvođači vina odbacuju neiskorišteni trop iako sadržava bioaktivne spojeve s antioksidacijskim i protuupalnim svojstvima. Svrha je ovoga rada bila ispitati antinociceptivna i protuupalna svojstva vodeno-etanolnog ekstrakta pokožice grožđa vrste Vitis labrusca iz agroindustrijskog otpada. Značaj je ovoga istraživanja u tome što se korištenjem otpada daje dodatna ekonomska vrijednost grožđu u uzgojnom lancu. Eksperimentalni pristup. Ekstrakt je dobiven iz pokožica izdvojenih iz reprezentativnog uzorka komine. Udjel fenolnih spojeva određen je praćenjem višestrukih reakcija pomoću masene spektrometrije i metodom Folin-Ciocalteu, uz galnu kiselinu kao standard. Biološka aktivnost ekstrakata ispitana je na miševima koji su hranjeni sirovim ekstraktom u dozama od 30, 100 i 300 mg/kg. Ispitani su sljedeći parametri: mehanička hiperalgezija pomoću von Frey filamenata, toplinska hiperalgezija na vrućoj ploči pri 55 °C, edem šape pomoću pomičnog mjerila i aktivnost mijeloperoksidaze kao pokazatelj aktivacije neutrofila. Nefrotoksičnost i hepatotoksičnost su ispitane biokemijskim pretragama uzoraka krvi miševa hranjenih ekstraktom grožđa vrste Vitis labrusca. Rezultati i zaključci. Maseni udjel pokožica u svim uzorcima otpada vinske industrije iznosio je 75 % mokre tvari i 59 % suhe tvari. Identificirali smo devet različitih antocijanina (3-O-glukozide peonidin, delfinidin, petunidin i malvidin; 3-p-kumaroil-glukozide cijanidin, peonidin, pe¬tunidin i malvidin, te malvidin-3,5-diglukozid), pet flavonoida (apigenin-7-glukozid, luteolin-7-glukozid, kvercetin-3-galaktozid, izorhamnetin-3-glukozid i miri¬cetin-3-rutinozid), a maseni udjel fenolnih spojeva, izražen kao ekvivalent galne kiseline, bio je 26,62 mg/g. Ispitivanja in vivo pokazala su da su ektrakti grožđa vrste Vitis labrusca masenog udjela 100 i 300 mg/kg smanjili mehaničku i toplinsku hiperalgeziju kod miševa nakon injekcije karagenana, reducirali edem šape za 50 % i smanjili broj neutrofila. Osim toga, nije bilo pokazatelja nefrotoksičnosti i hepatotoksičnosti. Ekstrakt dobiven iz otpada vinske industrije ima analgetska i protuupalna svojstva, djelomično zbog toga što sadržava fenolne spojeve, a nije toksičan za tkiva bubrega i jetre. Novina i znanstveni doprinos. Dobiveni se bioproizvod može upotrijebiti kao alternativa sintetičkim protuupalnim agensima, s istim farmakološkim potencijalom a manje nuspojava. Pokazali smo da se vinski otpad grožđa vrste Vitis labrusca može upotrijebiti za proizvodnju antinociceptivnih i protuupalnih proizvoda (nutraceutičkih, farmaceutskih i kozmetičkih) koji nemaju toksični učinak, te na taj način pridonijeti zaštiti okoliša

The Lipid Mediator Resolvin D1 Reduces the Skin Inflammation and Oxidative Stress Induced by UV Irradiation in Hairless Mice

UV irradiation-induced oxidative stress and inflammation contribute to the development of skin diseases. Therefore, targeting oxidative stress and inflammation might contribute to reduce skin diseases. Resolvin D1 (RvD1) is a bioactive metabolite generated during inflammation to actively orchestrate the resolution of inflammation. However, the therapeutic potential of RvD1 in UVB skin inflammation remains undetermined, which was, therefore, the aim of the present study. The intraperitoneal treatment with RvD1 (3-100 ng/mouse) reduced UVB irradiation-induced skin edema, myeloperoxidase activity, matrix metalloproteinase 9 activity, and reduced glutathione depletion with consistent effects observed with the dose of 30 ng/mouse, which was selected to the following experiments. RvD1 inhibited UVB reduction of catalase activity, and hydroperoxide formation, superoxide anion production, and gp91phox mRNA expression. RvD1 also increased the Nrf2 and its downstream targets NQO1 and HO-1 mRNA expression. Regarding cytokines, RvD1 inhibited UVB-induced production of IL-1β, IL-6, IL-33, TNF-α, TGF-β, and IL-10. These immuno-biochemical alterations by RvD1 treatment had as consequence the reduction of UVB-induced epidermal thickness, sunburn and mast cell counts, and collagen degradation. Therefore, RvD1 inhibited UVB-induced skin oxidative stress and inflammation, rendering this resolving lipid mediator as a promising therapeutic agent

Pectin/hydroxypropylmethylcellulose matrix tablets designed for colon-specific delivery containing Quercetin

O principal objetivo deste trabalho foi desenvolver sistemas matriciais obtidos da mistura pectina/ HPMC e lactose/HPMC como polímeros capazes de modular a liberação colônica de quercetina. Os comprimidos matriciais preparados utilizando-se os polímeros pectina e HPMC foram analisados pelo teste de dissolução in vitro, por período de 8 h. Os resultados demonstraram um período de latência de aproximadamente 4 h para início da liberação de quercetina, tempo necessário para que o fármaco alcance o cólon. Considerando os resultados, a formulação 2 (pectina/HPMC) foi capaz de resistir à liberação da quercetina em meio ácido, permitindo que o fármaco alcance o local de ação na concentração desejada. Desta forma, sistemas matriciais de liberação cólon-específica contendo quercetina podem ser uma estratégia promissora para o tratamento local da DII.The purpose of the study was to develop matrix systems acquired by the mixture pectin/HPMC and lactose/HPMC like polymers able to modulate the colonic release of quercetin. The matrix tablets prepared by using the polymers pectin and HPMC were analyzed through the in vitro dissolution test for eight hours. The results showed a latent period of approximately four hours before the beginning of quercetin release, the necessary time so that the drug reaches the colon. By considering the results, the formulation 2 (pectin/HPMC) was able to resist to the quercetin release in acid environment which allowed the drug to reach the action place in desired concentration. Thus, matrix systems of colon-specific release that contain quercetin can be a successful strategy to the local treatment of DII.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Pectin/hydroxypropylmethylcellulose matrix tablets designed for colon-specific delivery containing Quercetin

O principal objetivo deste trabalho foi desenvolver sistemas matriciais obtidos da mistura pectina/ HPMC e lactose/HPMC como polímeros capazes de modular a liberação colônica de quercetina. Os comprimidos matriciais preparados utilizando-se os polímeros pectina e HPMC foram analisados pelo teste de dissolução in vitro, por período de 8 h. Os resultados demonstraram um período de latência de aproximadamente 4 h para início da liberação de quercetina, tempo necessário para que o fármaco alcance o cólon. Considerando os resultados, a formulação 2 (pectina/HPMC) foi capaz de resistir à liberação da quercetina em meio ácido, permitindo que o fármaco alcance o local de ação na concentração desejada. Desta forma, sistemas matriciais de liberação cólon-específica contendo quercetina podem ser uma estratégia promissora para o tratamento local da DII.The purpose of the study was to develop matrix systems acquired by the mixture pectin/HPMC and lactose/HPMC like polymers able to modulate the colonic release of quercetin. The matrix tablets prepared by using the polymers pectin and HPMC were analyzed through the in vitro dissolution test for eight hours. The results showed a latent period of approximately four hours before the beginning of quercetin release, the necessary time so that the drug reaches the colon. By considering the results, the formulation 2 (pectin/HPMC) was able to resist to the quercetin release in acid environment which allowed the drug to reach the action place in desired concentration. Thus, matrix systems of colon-specific release that contain quercetin can be a successful strategy to the local treatment of DII.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Pectin/hydroxypropylmethylcellulose matrix tablets designed for colon-specific delivery containing Quercetin

O principal objetivo deste trabalho foi desenvolver sistemas matriciais obtidos da mistura pectina/ HPMC e lactose/HPMC como polímeros capazes de modular a liberação colônica de quercetina. Os comprimidos matriciais preparados utilizando-se os polímeros pectina e HPMC foram analisados pelo teste de dissolução in vitro, por período de 8 h. Os resultados demonstraram um período de latência de aproximadamente 4 h para início da liberação de quercetina, tempo necessário para que o fármaco alcance o cólon. Considerando os resultados, a formulação 2 (pectina/HPMC) foi capaz de resistir à liberação da quercetina em meio ácido, permitindo que o fármaco alcance o local de ação na concentração desejada. Desta forma, sistemas matriciais de liberação cólon-específica contendo quercetina podem ser uma estratégia promissora para o tratamento local da DII.The purpose of the study was to develop matrix systems acquired by the mixture pectin/HPMC and lactose/HPMC like polymers able to modulate the colonic release of quercetin. The matrix tablets prepared by using the polymers pectin and HPMC were analyzed through the in vitro dissolution test for eight hours. The results showed a latent period of approximately four hours before the beginning of quercetin release, the necessary time so that the drug reaches the colon. By considering the results, the formulation 2 (pectin/HPMC) was able to resist to the quercetin release in acid environment which allowed the drug to reach the action place in desired concentration. Thus, matrix systems of colon-specific release that contain quercetin can be a successful strategy to the local treatment of DII.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Antinociceptive Effect of Tephrosia sinapou Extract in the Acetic Acid, Phenyl-p-benzoquinone, Formalin, and Complete Freund’s Adjuvant Models of Overt Pain-Like Behavior in Mice

Tephrosia toxicaria, which is currently known as Tephrosia sinapou (Buc’hoz) A. Chev. (Fabaceae), is a source of compounds such as flavonoids. T. sinapou has been used in Amazonian countries traditional medicine to alleviate pain and inflammation. The purpose of this study was to evaluate the analgesic effects of T. sinapou ethyl acetate extract in overt pain-like behavior models in mice by using writhing response and flinching/licking tests. We demonstrated in this study that T. sinapou extract inhibited, in a dose (1–100 mg/kg) dependent manner, acetic acid- and phenyl-p-benzoquinone- (PBQ-) induced writhing response. Furthermore, it was active via intraperitoneal, subcutaneous, and peroral routes of administration. T. sinapou extract also inhibited formalin- and complete Freund’s adjuvant- (CFA-) induced flinching/licking at 100 mg/kg dose. In conclusion, these findings demonstrate that T. sinapou ethyl acetate extract reduces inflammatory pain in the acetic acid, PBQ, formalin, and CFA models of overt pain-like behavior. Therefore, the potential of analgesic activity of T. sinapou indicates that it deserves further investigation

Therapeutic Potential of Controlled Delivery Systems in Asthma: Preclinical Development of Flavonoid-Based Treatments

Asthma is a chronic disease with increasing prevalence and incidence, manifested by allergic inflammatory reactions, and is life-threatening for patients with severe disease. Repetitive challenges with the allergens and limitation of treatment efficacy greatly dampens successful management of asthma. The adverse events related to several drugs currently used, such as corticosteroids and β-agonists, and the low rigorous adherence to preconized protocols likely compromises a more assertive therapy. Flavonoids represent a class of natural compounds with extraordinary antioxidant and anti-inflammatory properties, with their potential benefits already demonstrated for several diseases, including asthma. Advanced technology has been used in the pharmaceutical field to improve the efficacy and safety of drugs. Notably, there is also an increasing interest for the application of these techniques using natural products as active molecules. Flavones, flavonols, flavanones, and chalcones are examples of flavonoid compounds that were tested in controlled delivery systems for asthma treatment, and which achieved better treatment results in comparison to their free forms. This review aims to provide a comprehensive understanding of the development of novel controlled delivery systems to enhance the therapeutic potential of flavonoids as active molecules for asthma treatment

Method validation and stability study of quercetin in topical emulsions

This study validated a high performance liquid chromatography (HPLC) method for the quantitative evaluation of quercetin in topical emulsions. The method was linear within 0.05 - 200 μg/mL range with a correlation coefficient of 0.9997, and without interference in the quercetin peak. The detection and quantitation limits were 18 and 29 ng/mL, respectively. The intra- and inter-assay precisions presented R.S.D. values lower than 2%. An average of 93% and 94% of quercetin was recovered for non-ionic and anionic emulsions, respectively. The raw material and anionic emulsion, but not non-ionic emulsion, were stable in all storage conditions for one year. The method reported is a fast and reliable HPLC technique useful for quercetin determination in topical emulsions

Quality Evaluation of Manipulated and Industrialized Simvastatin Formulations

Medicamentos de uso crônico, como a sinvastatina, podem representar ao usuário um custo elevado, propiciando a utilização de medicamentos manipulados como uma alternativa, no entanto a sua qualidade deve ser assegurada. Assim, o trabalho tem como objetivo verificar a qualidade de comprimidos referência (R), genéricos (G1-2) e similares (S1-2) e cápsulas manipuladas (M1-4) de sinvastatina avaliando- se peso médio, perfil de dissolução, doseamento e uniformidade de conteúdo. Os resultados da avaliação da qualidade dos comprimidos industrializados e das cápsulas manipuladas 1-3 estão de acordo com os parâmetros especificados nas Farmacopéias. No entanto, os resultados da uniformidade de conteúdo para a formulação M4 indicaram que 3 amostras apresentaram teor inferior a 75% com DPR de 10,30%. Ainda, a quantidade de sinvastatina liberada foi de 63,74% em 30 minutos não atendendo o recomendado pela Farmacopéia Americana. O trabalho mostrou a importância em assegurar a qualidade dos medicamentos comercializados com o interesse em evitar ineficácia terapêutica.Chronic drug treatments with simvastatin may be expensive to patients, favoring the use of manipulated pharmaceutical forms as an alternative, thus, quality must be ensured. Thus, the study addressed the quality of the reference (R), generics (G1 and G2) and similars (S1 and S2) industrialized tablets, and four pharmacy manipulated capsules (M1-4) of simvastatin by evaluating the average weight, dissolution profile, content of active substance and uniformity of dosage unity. All parameters tested were in accordance with the Pharmacopeias for all dosage forms, except for the uniformity of dosage unity of M4, which presented less than 75% of the expected simvastatin content with relative standard deviation of 10.30 % for three samples. Furthermore, the amount of released sinvastatin was 63.74% within 30 minutes, which is below the recommendation of the American Pharmacopeia. Thus, it was shown the importance to ensure the quality of commercially available medicines avoiding therapeutic inefficacy.Universidad Nacional de La Plat