Green Propolis: A Review of Pharmaceutical Patents with Potential Therapeutic Applications.

Green propolis is a resin produced from Baccharis dracunculifolia, which has as its main compounds flavonoids, derivatives of cinnamic acids, such as artepillin C and baccarin. This resin has antibacterial, antifungal, anti-inflammatory, antioxidant, and anticancer activities. This review aimed to analyze pharmaceutical patents containing green propolis in various formulations. The search was conducted in the National Institute of Industrial Property (INPI), the Patent Search of Spain and Latin America (Latipat-Espacenet), the World Intellectual Property Organization (WIPO), and Google Patents, with a combination of the keywords green propolis, green propolis extract, pharmacology, and pharmaceutical product. Primary research identified 60 patents, from which 22 were selected after applying the inclusion criteria. The selected patents referred to products with pharmacological activities, from cancer treatment to food supplements and included innovations for improved controlled release of the green propolis compounds. Most of the documents concerned the preparation and/or formulation of the green propolis extract, followed by innovative extraction methods, treatment and systemic use, and finally by topical use and quality control techniques and procedures. Thus, the reviewed patents of green propolis provided valuable insights into the pharmaceutical applications of green propolis, showing its potential in diverse formulations and treatments.

In vivo anti-inflammatory activity of BACCHARIN from BRAZILIAN green PROPOLIS.

Baccharin is one of the major compounds found in Brazilian green propolis and its botanical source, Baccharis dracunculifolia. Considering the biological effects of propolis and B. dracunculifolia, this study aims to evaluate the analgesic and anti-inflammatory potential of baccharin. The neurodepressor potential was performed by the open field test, analgesia by mechanical stimulation with Dynamic Plantar Aesthesiometer, and by thermal stimulation with Hargreaves apparatus. In addition, the anti-inflammatory potential was achieved by the paw edema assay, histopathological evaluation, and NF-kB expression. Doses of 2.5, 5, and 10 mg/kg of baccharin were evaluated. After euthanasia, plantar tissue was collected and prepared for histology. As a result, analgesic activity was observed at a dose of 10 mg/kg of baccharin in thermal stimulation under an inflammatory process and anti-inflammatory potential at a dose of 5 mg/kg of baccharin from the second hour in the paw edema test. A decrease in cellular infiltrate and down-modulation of NF-kB, besides the reduction of edema in the histopathology was observed. There was no evidence of kidney and liver toxicity and neurodepressive potential at the doses tested. Thus, baccharin has a promising anti-inflammatory effect possibly associated with antiedematogenic activity by inhibiting mediators such as prostaglandins, inhibiting the migration of polymorphonuclear cells, and modulating NF-kB expression.

Baccharis dracunculifolia DC Consumption Improves Nociceptive and Depressive-like Behavior in Rats with Experimental Osteoarthritis.

Osteoarthritis (OA) persistently activates nociceptors, leading to chronic pain, which is often accompanied by the comorbid development of emotional impairments (anxiety and depression), an effect associated with microgliosis. Baccharis dracunculifolia DC (Asteraceae), a Brazilian edible plant, is an important source of active compounds with anti-inflammatory abilities. Thus, we evaluated its ability to reverse OA-induced nociceptive and emotional-like impairments in osteoarthritic ovariectomized female rats using the kaolin/carrageenan (K/C) model. Four weeks after OA induction, mechanical hyperalgesia was confirmed, and the treatment started. Control animals (SHAMs) were treated with phosphate-buffered saline (PBS), while arthritic animals (ARTHs) either received PBS or B. dracunculifolia 50 mg/kg (Bd50) and 100 mg/kg (Bd100), via gavage, daily for five weeks. At the end of the treatment, anxiety-like behavior was assessed using the Open Field Test (OFT), anhedonia was assessed using the Sucrose Preference Test (SPT), and learned helplessness was assessed using the Forced Swimming Test (FST). After occision, microglia were stained with IBA-1 and quantified in brain sections of target areas (prefrontal cortex, amygdala, and periaqueductal grey matter). Treatment with B. dracunculifolia extract reversed OA-induced mechanical hyperalgesia and partly improved depressive-like behavior in OA animals' concomitant to a decrease in the number of M1 microglia. Our findings suggest that B. dracunculifolia extracts can potentially be used in the food industry and for the development of nutraceuticals and functional foods.

Farnesol, a component of plant-derived honeybee-collected resins, shows JH-like effects in Apis mellifera workers.

Farnesol, a sesquiterpene found in all eukaryotes, precursor of juvenile hormone (JH) in insects, is involved in signalling, communication, and antimicrobial defence. Farnesol is a compound of floral volatiles, suggesting its importance in pollination and foraging behaviour. Farnesol is found in the resin of Baccharis dracunculifolia, from which honeybees elaborate the most worldwide marketable propolis. Bees use propolis to seal cracks in the walls, reinforce the wax combs, and as protection against bacteria and fungi. The introduction within a honeybee hive of a compound with potential hormonal activity can be a challenge to the colony survival, mainly because the transition from within-hive to outside activities of workers is controlled by JH. Here, we tested the hypothesis that exogenous farnesol alters the pacing of developing workers. The first assays showed that low doses of the JH precursor (0.1 and 0.01 µg) accelerate pharate-adult development, with high doses being toxic. The second assay was conducted in adult workers and demonstrated bees that received 0.2 µg farnesol showed more agitated behaviour than the control bees. If farnesol was used by corpora allata (CA) cells as a precursor of JH and this hormone was responsible for the observed behavioural alterations, these glands were expected to be larger after the treatment. Our results on CA measurements after 72 h of treatment showed bees that received farnesol had glands doubled in size compared to the control bees (p < 0.05). Additionally, we expected the expression of JH synthesis, JH degradation, and JH-response genes would be upregulated in the treated bees. Our results showed that indeed, the mean transcript levels of these genes were higher in the treated bees (significant for methyl farnesoate epoxidase and juvenile hormone esterase, p < 0.05). These results suggest farnesol is used in honeybees as a precursor of JH, leading to increasing JH titres, and thus modulating the pacing of workers development. This finding has behavioural and ecological implications, since alterations in the dynamics of the physiological changes associated to aging in young honeybees may significantly impact colony balance in nature.

Experimental evidence for a hidden network of higher-order interactions in a diverse arthropod community.

Transcending pairwise interactions in ecological networks remains a challenge.1,2,3,4,5 Higher-order interactions (HOIs), the modulation of a pairwise interaction by a third species,6 are thought to play a particularly important role in stabilizing coexistence and maintaining species diversity.7,8,9,10,11,12 However, HOIs have so far only been demonstrated in models9,10,11,12,13,14 or isolated experimental systems including only a few interacting species.7,8,15 Their ubiquity and importance at a community level in the real world remain unknown. We hypothesized that a complex network of HOIs could be constantly modifying pairwise interactions and shaping ecological communities and that consequently the outcome of pairwise interactions would be a product of many influences from distinct sources. Using field experiments, we tested how multiple interactions within a diverse arthropod community associated with the tropical shrub Baccharis dracunculifolia D.C. (Asteraceae) were modified by the removal of ant species or live or hatched insect galls (a non-trophic engineering effect) of the dominant galler species. We revealed an extensive hidden network of HOIs modifying each other and the "visible" pairwise interactions. Most pairwise interactions were affected indirectly by the manipulation of non-interacting taxonomic groups. The pervasiveness of these interaction modifications challenges pairwise approaches to understanding interaction outcomes and could shift our thinking about the structure and persistence of ecological communities. Investigating coexistence mechanisms involving interaction modulation by HOIs may be key to elucidating the underlying causes of the stability and persistence of ecological communities.

Chemical composition, antifungal and antioxidant activity of essential oils from Baccharis dracunculifolia and Pogostemon cablin against Fusarium graminearum.

Fusarium graminearum Schw. is associated with diseases in seeds and seedlings, mainly in maize, and causes damage to crops and food production. We determined the chemical compositions of essential oils from Pogostemon cablin (Blanco) Benth and Baccharis dracunculifolia DC., and evaluated their antioxidant and antifungal activity against F. graminearum. The oils were applied to the fungus using disc diffusion technique at concentrations of 8.0, 4.0, 2.0 and 1.0 µL mL-1. Fungistatic effects of P. cablin oil were detectable at 8.0 µL mL-1, with 80.0% inhibition of fungal growth. This oil contains mono and sesquiterpenes that may be toxic to fungal cell structures. P. cablin oil also had antioxidant activity in free radical sequestration experiments, where as oil from B. dracunculifolia had limited effects. We conclude that essential oil from P. cablin has greater anti-phytopathogenic and antioxidant activity than that from B. dracunculifolia.

How diverse is the chemistry and plant origin of Brazilian propolis?

Propolis is a honey bee product containing chiefly beeswax and resins originated from plant buds or exudates. Propolis resin exerts a diversity of biological activities, such as antitumoral, anti-inflammatory, antimicrobial, and defense of the hive against pathogens. Chemical standardization and identification of botanical sources is crucial for characterization of propolis. Types of Brazilian propolis are characteristic of geographical regions and respective biomes, such as savannas (Cerrado), mangroves, dry forest (Caatinga), rain forests (Amazon, Atlantic, and Interior forests), altitudinal fields ("Campos Rupestres"), Pantanal, and Araucaria forests. Despite the wide diversity of Brazilian biomes and flora, relatively few types of Brazilian propolis and corresponding resin plant sources have been reported. Factors accounting for the restricted number of known types of Brazilian propolis and plant sources are tentatively pointed out. Among them, the paper discusses constraints that honey bees must overcome to collect plant exudates, including the characteristics of the lapping-chewing mouthpart of honey bee, which limit their possibilities to cut and chew plant tissues, as well as chemical requirements that plant resins must fulfil, involving antimicrobial activity of its constituents and innocuity to the insects. Although much still needs to be done toward a more comprehensive picture of Brazilian propolis types and corresponding plant origins, the prospects indicate that the actual diversity of plant sources of honey bee propolis will remain relatively low.

Synthesis, antitumor activity and in silico analyses of amino acid derivatives of artepillin C, drupanin and baccharin from green propolis.

Breast cancer has the highest incidence and mortality in females, while prostate cancer has the second-highest incidence in males. Studies have shown that compounds from Brazilian green propolis have antitumor activities and can selectively inhibit the AKR1C3 enzyme, overexpressed in hormone-dependent prostate and breast tumors. Thus, in an attempt to develop new cytotoxic inhibitors against these cancers, three prenylated compounds, artepillin C, drupanin and baccharin, were isolated from green propolis to synthesize new derivatives via coupling reactions with different amino acids. All obtained derivatives were submitted to antiproliferative assays against four cancer cells (MCF-7, MDA MB-231, PC-3, and DU145) and two normal cell lines (MCF-10A and PNT-2) to evaluate their cytotoxicity. In general, the best activity was observed for compound6e, derived from drupanin, which exhibited half-maximal inhibitory concentration (IC50) of 9.6 ± 3 µM and selectivity index (SI) of 5.5 against MCF-7 cells.In silicostudies demonstrated that these derivatives present coherent docking interactions and binding modes against AKR1C3, which might represent a possible mechanism of inhibition in MCF-7 cells.

Antimicrobial activity of Baccharis dracunculifolia DC and its synergistic interaction with nisin against food-related bacteria.

The antimicrobial activities of Baccharis dracunculifolia DC essential oil (EO) and hydroalcoholic extract (HE) were evaluated. The EO showed broad antimicrobial activity and its synergistic combination with nisin was tested. Major components of EO were nerolidol, beta-pinene and D-limonene, while artepillin C, rutin and cafeic acid were major phenolics of HE. EO and HE were tested by agar diffusion assay against several strains of bacteria and yeasts, and mixed cultures of bacterial strains. The EO presented the largest spectrum of antimicrobial activity inhibiting all Gram-positive bacteria tested. Yeasts were not inhibited. The effect of EO against mixtures of sensitive and non-sensitive bacteria was tested on milk agar, being the inhibitory effect only observed on mixtures containing susceptible strains. The combination of EO and nisin at ½ MIC was evaluated on the growth curve of Staphylococcus aureus, Bacillus cereus, Listeria monocytogenes and Salmonella Enteritidis during 24 h at 37 °C. The combination EO-nisin was effective and no viable counts of B. cereus, L. monocytogenes and S. Enteritidis was observed, while the individual antimicrobials caused no inhibition. The counts of S. aureus were about 4 log CFU/mL lower in comparison with EO or nisin alone. B. dracunculifolia DC may be a potential source of natural antimicrobials, and its synergistic effect with nisin would reduce the working concentration, minimizing the organoleptic effects associated with this plant antimicrobial.

Baccharin and p-coumaric acid from green propolis mitigate inflammation by modulating the production of cytokines and eicosanoids.

ETHNOPHARMACOLOGICAL RELEVANCE: Green propolis is produced by Apis mellifera honeybees using Baccharis dracunculifolia D.C. (Asteraceae) as substrate. This Southern Brazilian native plant and green propolis have been used in traditional medicine to treat gastric diseases, inflammation and liver disorders. AIM OF THE STUDY: Investigate the effects of baccharin (Bac) or p-coumaric acid (pCA) isolated from B. dracunculifolia D.C. (Asteraceae) over the inflammation induced by lipopolysaccharide (LPS) in vivo. MATERIALS AND METHODS: Inflammation was induced by LPS injection into air-pouches in mice, which were subsequently treated with Bac or pCA. Lavage fluid was collected from air pouches for the quantification of cellular influx via microscopy, and quantification of inflammatory mediators via colorimetric methods, ELISA and liquid chromatography-tandem mass spectrometry (LC-MS/MS). RESULTS: LPS-induced inflammation increased cellular influx and increased the levels of parameters related to vascular permeability and edema formation, such as nitric oxide (NO) and protein extravasation. Moreover, LPS increased the levels of cytokines and eicosanoids in the air-pouches. Importantly, both Bac and pCA suppressed the infiltration of neutrophils, production of NO and protein extravasation. Notably, the compounds promote differential regulation of cytokine and eicosanoid production. CONCLUSIONS: Our results suggest that Bac from green propolis directly affects inflammation by inhibiting the production of cytokines and eicosanoids, while pCA may exert direct, but also indirect effects on inflammation by stimulating the production of regulatory effectors such as interkeukin-10 in vivo.

Relationship between Baccharis dracunculifolia DC. and B. microdonta DC. (Asteraceae) by Their Different Seasonal Volatile Expression.

Baccharis dracunculifolia DC. and Baccharis microdonta DC. (Asteraceae) are woody species morphologically similar growing in Uruguay, where not taxonomists people often confuse them in field conditions. As the essential oil of B. dracunculifolia ('vassoura' oil) is highly prized by the flavor and fragrance industry, the correct differentiation of the two species is a key factor in exploiting them profitably and reasonably. To differentiate both Baccharis species, in this work their volatile expression profiles were studied as an alternative tool to determine authenticity and quality. Volatile organic compounds (VOCs) were monthly extracted during an entire year from aerial parts of wild populations by simultaneous distillation extraction (SDE), and studied by gas chromatography/mass spectrometry (GC/MS; identification) and conventional gas chromatography (GC-FID; component abundances determination). Enantioselective gas chromatography/mass spectrometry (Es-GC/MS) was applied in the search of parameters able to ensure genuineness of each species extract. Qualitative VOCs profiles were found to be similar for both species, being ß-pinene, limonene, spathulenol, caryophyllene oxide, and viridiflorol the main components. However, the abundance of those VOCs were two to ten times higher in B. dracunculifolia than in B. microdonta during the year of study. These Baccharis spp. showed species-specific patterns of VOCs expression according to the seasonality, and interestingly, oxygenated compounds (trans-pinocarveol and myrtenal) increased their abundances at full-flowering stages. The enantiomeric distribution of selected monoterpenes (α- and ß-pinenes, limonene, linalool, terpinen-4-ol, and α-terpineol) presented differential values for both Baccharis spp., meaning that Es-GC might be a useful tool for differentiating chemically both species in Uruguay for genuineness determination purposes.

Artepillin C: A comprehensive review of its chemistry, bioavailability, and pharmacological properties.

Artepillin C (ARC), a prenylated derivative of p-coumaric acid, is one of the major phenolic compounds found in Brazilian green propolis (BGP) and its botanical source Baccharis dracunculifolia. Numerous studies on ARC show that its beneficial health effects correlate with the health effects of both BGP and B. dracunculifolia. Its wide range of pharmacological benefits include antioxidant, antimicrobial, anti-inflammatory, anti-diabetic, neuroprotective, gastroprotective, immunomodulatory, and anti-cancer effects. Most studies have focused on anti-oxidation, inflammation, diabetic, and cancers using both in vitro and in vivo approaches. Mechanisms underlying anti-cancer properties of ARC are apoptosis induction, cell cycle arrest, and the inhibition of p21-activated kinase 1 (PAK1), a protein characterized in many human diseases/disorders including COVID-19 infection. Therefore, further pre-clinical and clinical studies with ARC are necessary to explore its potential as intervention for a wide variety of diseases including the recent pandemic coronaviral infection. This review summarizes the comprehensive data on the pharmacological effects of ARC and could be a guideline for its future study and therapeutic usage.

Baccharis dracunculifolia and Dalbergia ecastophyllum, Main Plant Sources for Bioactive Properties in Green and Red Brazilian Propolis.

Nowadays, propolis is used as a highly valuable product in alternative medicine for improving health or treating a large spectrum of pathologies, an ingredient in pharmaceutical products, and also as a food additive. Different vegetal materials are collected by honeybees and mixed with wax and other own substances in order to obtain the final product, called propolis. It is known as the bee product with the widest chemical composition due to the raw material collected by the bees. Different types are known worldwide: green Brazilian propolis (having Baccharis dracunculifolia as the major plant source), red Brazilian propolis (from Dalbergia ecastophyllum), European propolis (Populus nigra L.), Russian propolis (Betula verrucosa Ehrh), Cuban and Venezuelan red propolis (Clusia spp.), etc. An impressive number of scientific papers already demonstrate the pharmacological potential of different types of propolis, the most important activities being the antimicrobial, anti-inflammatory, antitumor, immunomodulatory, and antioxidant activities. However, the bioactive compounds responsible for each activity have not been fully elucidated. This review aims to collect important data about the chemical composition and bioactive properties of the vegetal sources and to compare with the chemical composition of respective propolis types, in order to determine the connection between the floral source and the propolis properties.

Chemical composition and antimycoplasma activity of a brown propolis from southern Brazil.

Extracts of a sample of brown propolis produced in the district of Itapará (Southern Brazil) were obtained with solvents with increasing polarity. The extracts were analyzed by RPHPLC-DAD-ESI-MS/MS and evaluated toward activity against Mycoplasma bovis, M. gallisepticum, M. genitalium, M. hominis, M. hyorinis, M. penetrans and M. pneumonieae. Typical components of "alecrim-do-campo" propolis (e.g. prenylated phenylpropanoids and caffeoyl-quinic acids) were characterized in the analyzed extraccts, in addition to several flavonols. Less polar extracts showed higher anti-mycoplasma activity (MIC value commonly 3.9 µg/mL) than alcoholic and aqueous extracts (MIC value often 7.8-250 µg/mL). The results indicate that Itapará propolis is a promising source for the development of therapeutic drugs.

Baccharis dracunculifolia (Asteraceae) essential oil displays anti-inflammatory activity in models of skin inflammation.

ETHNOPHARMACOLOGICAL RELEVANCE: Baccharis dracunculifolia (Asteraceae) is a commonly used plant in traditional medicine known as "alecrim-do-campo". Popularly it has been used as an immunostimulant, antibiotic, anti-inflammatory among other applications. So far, only a few studies have investigated the B. dracunculifolia anti-inflammatory effect and none has investigated the effectiveness of essential oil on skin diseases. AIM OF THE STUDY: The study aimed at evaluating the topical anti-inflammatory activity of B. dracunculifolia essential oil (BdEO) in mice models of acute and chronic skin inflammation. MATERIALS AND METHODS: BdEO was obtained from leaves and it was analyzed with Gas Chromatograph. Topical anti-inflammatory activity of BdEO (0.1, 0.3 and 1.0 mg/ear) was evaluated in Arachidonic Acid or TPA-induced acute and chronic skin inflammation in mice. Parameters such edema, cell migration and keratinocytes proliferation were evaluated. In addition, safety and a possible mechanism of action for BdEO essential oil were also investigated. RESULTS: Our results indicate that mainly terpenoids compounds compose BdEO. In addition, topical treatment with BdEO inhibited inflammatory parameters in both acute and chronic models of skin inflammation. This protective effect was associated with reduced edema formation, smaller cellular influx into the inflamed tissue and reduction of keratinocytes hyperproliferation. Although BdEO appears to exert its anti-inflammatory effect through a corticosteroid pathway, no local or systemic side effects were observed. CONCLUSION: Taken together, the present results showed that the essential oil obtained by hydrodistillation from B. dracunculifolia leaf samples exhibit remarkable topical anti-inflammatory properties. Therefore, our study demonstrated evidence for BdEO topical anti-inflammatory efficacy and safety, suggesting that it could be considered for developing of a new phytotherapeutic formulation as treatment for skin diseases.

Efficacy of Baccharis dracunculifolia in the treatment of diversion colitis in rats / Eficácia da Baccharis dracunculifolia no tratamento da colite de desuso em ratos

Abstract Rationale: Disuse colitis is frequent in our country and the most effective treatment is high cost and there is a need for effective and low cost therapy. Objective: To evaluate the efficacy of Baccharis dracunculifolia (field rosemary) in the treatment of exclusion colitis in rats. Method: Eighteen Wistar rats were anesthetized and submitted to colostomy; they were then distributed into two groups: Control Group, receiving intrarectal saline infusion (n = 8) and Group BD receiving intrarectal infusion ofBaccharis dracunculifolia extract (n = 10); after 21 days of treatment they were euthanized, the intestinal segment excluded from intestinal transit was resected and submitted to histopathological study, classifying the degree of inflammation and degree of vascular congestion from 0 to 3. Results: Mean inflammation was 2.7 in Control Group versus 2.1 in BD Group (p = 0.049), while mean vascular congestion was 2.3 and 2, respectively, in Control and BD groups (p = 0.1642). Conclusion: Intra-rectal infusion ofBaccharis dracunculifolia extract significantly minimized the inflammatory process in the exclusion colitis of rats submitted to colostomy, without altering the degree of vascular congestion.

Resumo Racional A colite de desuso é frequente em nosso meio e o tratamento de maior eficácia é de alto custo, havendo necessidade de se encontrar uma terapêutica eficaz e de baixo custo. Objetivo Avaliar a eficácia da Baccharis dracunculifolia (alecrim-do-campo) no tratamento da colite de exclusão em ratos. Método Utilizou-se 18 ratos Wistar, os quais foram anestesiados e submetidos à colostomia; em seguida distribuídos em 2 grupos: Grupo Controle, recebendo infusão intrarretal de solução salina (n = 8) e Grupo BD, recebendo infusão intrarretal de extrato de Baccharis dracunculifolia (n = 10); após 21 dias de tratamento foram submetidos a eutanásia, o segmento intesinal excluso de trânsito intestinal foi ressecado e submetido a estudo histopatológico classificando-se o grau de inflamação e grau de congestão vascular de 0 a 3. Resultados Verificou-se média de inflamação 2,7 no Grupo Controle vs. 2,1 no Grupo BD (p = 0,049), enquanto as médias de congestão vascular foram 2,3 e 2, respectivamente, nos grupos controle e BD (p = 0,1642). Conclusão A infusão intrarretal do extrato de Baccharis dracunculifolia minimizou significantemente o processo inflamatório na colite de exclusão de ratos submetidos à colostomia, sem alterar o grau de congestão vascular.

The Role of Baccharis dracunculifolia and its Chemical Profile on Green Propolis Production by Apis mellifera.

The botanical source of Brazilian green propolis (BGP) is Baccharis dracunculifolia DC, which interacts not only with Apis mellifera, but also with galling insects. In the last decade, because of green propolis´ important biologic activities, the international demand for BGP overcame the production capacity, consequently, new approaches are required to increase this production. Hence, the understanding of the chemical ecology interactions of B. dracunculifolia with galls and bees in field conditions may provide insights to increase BGP's production. A "bee pasture" experiment aiming to better understand this plant-insect interaction was therefore performed. For that, 48 B. dracunculifolia individuals, being 24 females and 24 males, were cultivated and investigated for the following parameters: (1) phenolic and volatile compounds in both B. dracunculifolia leaves and green propolis, (2) environmental variables, (3) visiting rate by bees, (4) time of resin collection, and (5) number of galls. Regression analyses by independent linear mixed-effect models were run to correlate phenolic and volatile compounds concentration with the environmental and field variables. Significant differences in chemical profile and field variables were observed between male and female plants. Male plants showed higher infestation by galling insects while female plants showed higher number of visiting bees, time of resin collection and terpenes concentration, contributing to the differences observed in the field. The obtained results suggest that increasing the percentage of female B. dracunculifolia plants in the field may attract more bees and therefore enhance propolis production.

Isolation, Identification, and Synthesis of a New Prenylated Cinnamic Acid Derivative from Brazilian Green Propolis and Simultaneous Quantification of Bioactive Components by LC-MS/MS.

A new cinnamic acid derivative, (E)-3-[4-hydroxy-3-((E)-3-formyl-2-butenyl)phenyl]-2- propenoic acid (20) has been isolated from the ethanol extract of Brazilian green propolis along with three known cinnamic acid derivatives, 3,4-dihydroxy-5-prenyl-(E)-cinnamic acid (4), capillartemisin A (6), and 2,2-dimethylchromene-6-(E)-propenoic acid (8), and a flavonoid, dihydrokaempferide (16) by liquid-liquid participation, a series of column chromatography and preparative HPLC. Their structures have been determined by spectroscopic analyses and chemical synthesis of compound 20. The simultaneous quantification of 20 constituents, including 10 cinnamic acid derivatives, 7 flavonoids, and 3 caffeoylquinic acid derivatives, has also been developed and validated using LC-MS/MS. The new compound 20 was shown to activate PPAR α but not PPAR ß or γ.

Natural deep eutectic solvents and aqueous solutions as an alternative extraction media for propolis.

Ethanolic extracts of propolis are consumed for their health benefits even though direct consumption of alcoholic extracts is not always ideal. Natural Deep Eutectic Solvents (NADES) can potentially extract similar compounds as alcoholic extracts while being better for direct consumption. Therefore, in this work alternative solvents for the extraction of green propolis including its biomarker artepillin C were examined. Sixteen NADES made from low toxicity chemicals, including the essential amino acid l-lysine, were explored along with twelve individual NADES components and honey, which showed similar physical-chemical properties to NADES. At 50 °C NADES made from choline chloride-propylene glycol or lactic acid proved to be equal or better than the benchmark EtOH:Water 7:3 (v/v). Alternatively, aqueous l-lysine appeared as a potential solvent for the preparation of aqueous propolis extracts. From these findings NADES, honey and aqueous l-lysine solutions all demonstrated the potential to replace ethanol or water for extracting green propolis.

Hydroalcoholic extract from Baccharis dracunculifolia recovers the gastric ulcerated tissue, and p-coumaric acid is a pivotal bioactive compound to this action.

Baccharis dracunculifolia is a medicinal plant native to southeastern Brazil and is the main botanical source used by bees (Apis mellifera) in the manufacture of green propolis and display similar gastroprotective action and chemical profile. This article reports the healing gastric ulcer activity of the hydroethanolic extract of B. dracunculifolia (HEBD) in an acetic acid-induced ulcer model. In addition to the extract, the isolated compounds ferulic acid, p-coumaric acid, caffeic acid, baccharin, and aromadendrin-4'-O-methyl ether were also assayed. HEBD at a dose of 300 mg/kg reduced the ulcerated area by 49.4% after treatment for 7 days, twice a day. Histological analyses revealed that the margins and base of the ulcer obtained significant regeneration, and periodic acid Schiff base staining showed a 78.2% increase in the mucin levels. The action on the enzymatic antioxidant system demonstrated an increased activity of superoxide dismutase and glutathione-S-transferase, in addition to raising glutathione reduced levels and myeloperoxidase activity. HEBD did not show cytotoxicity in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylterazole bromine test. In vitro, HEBD inhibited the H+ /K+ -ATPase enzyme and showed antioxidant activity in the 2,2 diphenyl-1-picryllydrazyl test. Regarding the isolated compounds, oral administration of p-coumaric acid (15 mg/kg) reduced the ulcerated area by 66.2%. The results suggest that HEBD recovers the gastric ulcerated tissue, raising mucus and antioxidant enzyme levels, and reducing the H+ /K+ -ATPase activity. In addition, the findings confirm that p-coumaric acid is a pivotal bioactive compound on the gastric healing effects elicited by HEBD. © 2019 BioFactors, 45(3):479-489, 2019.