Antinociceptive and in vitro anticancer activity of microencapsulated extracts and vouacapans isolated from Pterodon pubescens Benth

Orientadores: Mary Ann Foglio, João Ernesto de Carvalho, Rodney Alexandre Ferreira, RodriguesDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: As plantas medicinais têm sido fonte de agentes medicinais há milhares de anos para o tratamento de um amplo espectro de doenças. Entre as alternativas que podem ser utilizadas para preservar as características do material vegetal, destaca-se a encapsulação em micropartículas. Estudos prévios com espécie Pterodon pubescens Benth. (sucupira) avaliaram a eficácia (ação antiinflamatória e antinociceptiva) e a segurança (eventuais efeitos toxicológicos) dos extratos, frações ativas e compostos isolados de P. pubescens. No presente trabalho, avaliou-se o benefício do processo de microencapsulação por spray drying para preservação das características químicas, aliado a ações biológicas in vitro e in vivo dos extratos e compostos isolados. Extratos brutos produzidos com sementes provenientes de Minas Gerais (EBMG), Bahia (EBBA), Mato Grosso (EBMT) ou São Paulo (EBSP) apresentaram perfil químico distinto, demonstrando a variabilidade química da espécie de diferentes procedências. A partir dos extratos EBMG e EBSP foram isolados os vouacapanos C1 e C2, ambos com atividade antinociceptiva, sendo que a atividade de C1 esteve mais relacionada a componente de dor neurogênica. C1 e os extratos EBBA, EBSP e EBMT foram microencapsulados por spray drying e, juntamente com as amostras livres, armazenados em câmara climática com umidade e temperatura controladas para o estudo de estabilidade. Os resultados dos ensaios de atividade anticâncer in vitro em células tumorais humanas e atividade antinociceptiva em modelos com camundongos, demonstraram que a técnica de microencapsulação por spray drying preservou a ação, tanto do composto isolado quanto dos extratos demonstrando ser uma alternativa útil para prolongar o tempo de estocagem, principalmente do extrato bruto.As vantagens apresentadas pelo processo de microencapsulação dos produtos obtidos a partir de P. pubescens, motivam a continuidade dos estudos de padronização com esta espécie, para viabilizar um produto que atenda a parâmetros de reprodutibilidade, eficácia e segurança.Abstract: Over the years natural products, especially plants, have demonstrated to play a dominant role for drug discovery to treat human diseases. Micro encapsulation is an important approach to preserve natural products characteristics. Previous studies with Pterodon pubescens Benth., (sucupira) evaluated the efficacy (anti-inflammatory and antinociceptive) and safety (possible toxicological effects) of the extracts, active fractions and compounds isolated from P. pubescens. In the present study we evaluated the benefit of the microencapsulation process by spray drying to preserve the chemical characteristic monitored by in vitro and in vivo biological activities. Crude extracts obtained from Minas Gerais (EBMG), Bahia (EBBA), Mato Grosso (EBMT) or São Paulo (EBSP) seeds showed different quimical profiles. Vouacapans C1 and C2, isolated to EBMG and EBSP crude extracts, demonstrated antinociceptive activity, C1 activity related to neurogenic pain. The crude extracts EBBA, EBSP, EBMT and compound C1 were microencapsulated by spray drying and, along with control samples, stored in a climatic chamber with controlled humidity and temperature for the accelerated stability studies. The test results of in vitro anticancer activity in human tumor cells and antinociceptive activity in mice models, showed that the microencapsulation technique by spray drying preserve both the isolated compound and extracts. This technique is a useful alternative to extend storage time, especially of the crude extract. The advantages presented by P. pubescens microencapsulated products prompt to continue studies with this species, for the production of a product that meets reproductively, efficacy and safety parameters.MestradoFarmacologia, Anestesiologia e TerapeuticaMestre em Odontologi

Chitosan-tripolyphosphate Nanoparticles As Arrabidaea Chica Standardized Extract Carrier: Synthesis, Characterization, Biocompatibility, And Antiulcerogenic Activity

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Natural products using plants have received considerable attention because of their potential to treat various diseases. Arrabidaea chica (Humb. & Bonpl.) B. Verlot is a native tropical American vine with healing properties employed in folk medicine for wound healing, inflammation, and gastrointestinal colic. Applying nanotechnology to plant extracts has revealed an advantageous strategy for herbal drugs considering the numerous features that nanostructured systems offer, including solubility, bioavailability, and pharmacological activity enhancement. The present study reports the preparation and characterization of chitosan-sodium tripolyphosphate nanoparticles (NPs) charged with A. chica standardized extract (AcE). Particle size and zeta potential were measured using a Zetasizer Nano ZS. The NP morphological characteristics were observed using scanning electron microscopy. Our studies indicated that the chitosan/sodium tripolyphosphate mass ratio of 5 and volume ratio of 10 were found to be the best condition to achieve the lowest NP sizes, with an average hydrodynamic diameter of 150 +/- 13 nm and a zeta potential of +45 +/- 2 mV. Particle size decreased with AcE addition (60 +/- 10.2 nm), suggesting an interaction between the extract's composition and polymers. The NP biocompatibility was evaluated using human skin fibroblasts. AcE-NP demonstrated capability of maintaining cell viability at the lowest concentrations tested, stimulating cell proliferation at higher concentrations. Antiulcerogenic activity of AcE-NP was also evaluated with an acute gastric ulcer experimental model induced by ethanol and indomethacin. NPs loaded with A. chica extract reduced the ulcerative lesion index using lower doses compared with the free extract, suggesting that extract encapsulation in chitosan NPs allowed for a dose reduction for a gastroprotective effect. The AcE encapsulation offers an approach for further application of the A. chica extract that could be considered a potential candidate for ulcer-healing pharmaceutical systems.1038973909Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)FAPESP [2009/16484-1, 2010/07781-0, 2009/51020-6, 2011/09127-8

Chitosan-tripolyphosphate Nanoparticles As Arrabidaea Chica Standardized Extract Carrier: Synthesis, Characterization, Biocompatibility, And Antiulcerogenic Activity.

Natural products using plants have received considerable attention because of their potential to treat various diseases. Arrabidaea chica (Humb. & Bonpl.) B. Verlot is a native tropical American vine with healing properties employed in folk medicine for wound healing, inflammation, and gastrointestinal colic. Applying nanotechnology to plant extracts has revealed an advantageous strategy for herbal drugs considering the numerous features that nanostructured systems offer, including solubility, bioavailability, and pharmacological activity enhancement. The present study reports the preparation and characterization of chitosan-sodium tripolyphosphate nanoparticles (NPs) charged with A. chica standardized extract (AcE). Particle size and zeta potential were measured using a Zetasizer Nano ZS. The NP morphological characteristics were observed using scanning electron microscopy. Our studies indicated that the chitosan/sodium tripolyphosphate mass ratio of 5 and volume ratio of 10 were found to be the best condition to achieve the lowest NP sizes, with an average hydrodynamic diameter of 150±13 nm and a zeta potential of +45±2 mV. Particle size decreased with AcE addition (60±10.2 nm), suggesting an interaction between the extract's composition and polymers. The NP biocompatibility was evaluated using human skin fibroblasts. AcE-NP demonstrated capability of maintaining cell viability at the lowest concentrations tested, stimulating cell proliferation at higher concentrations. Antiulcerogenic activity of AcE-NP was also evaluated with an acute gastric ulcer experimental model induced by ethanol and indomethacin. NPs loaded with A. chica extract reduced the ulcerative lesion index using lower doses compared with the free extract, suggesting that extract encapsulation in chitosan NPs allowed for a dose reduction for a gastroprotective effect. The AcE encapsulation offers an approach for further application of the A. chica extract that could be considered a potential candidate for ulcer-healing pharmaceutical systems.103897-390

Riverhood: political ecologies of socionature commoning and translocal struggles for water justice

[EN] Mega-damming, pollution and depletion endanger rivers worldwide. Meanwhile, modernist imaginaries of ordering `unruly waters and humans' have become cornerstones of hydraulic-bureaucratic and capitalist development. They separate hydro/social worlds, sideline river-commons cultures, and deepen socio-environmental injustices. But myriad new water justice movements (NWJMs) proliferate: rooted, disruptive, transdisciplinary, multi-scalar coalitions that deploy alternative river-society ontologies, bridge South-North divides, and translate river-enlivening practices from local to global and vice-versa. This paper's framework conceptualizes `riverhood' to engage with NWJMs and river commoning initiatives. We suggest four interrelated ontologies, situating river socionatures as arenas of material, social and symbolic co-production: `river-as-ecosociety', `river-as-territory', `river-as-subject', and `river-as-movement'. globalThis work was supported by the ERC European Research Council under the European Union's Horizon 2020 research and innovation programme [Riverhood, Grant Number 101002921]; see also www.movingrivers.org.Boelens, R.; Escobar, A.; Bakker, K.; Hommes, L.; Swyngedouw, E.; Hogenboom, B.; Huijbens, EH.... (2023). Riverhood: political ecologies of socionature commoning and translocal struggles for water justice. The Journal of Peasant Studies. 50(3):1125-1156. https://doi.org/10.1080/03066150.2022.21208101125115650

Changes in plasma biomarkers following treatment with cabozantinib in metastatic castration-resistant prostate cancer: a post hoc analysis of an extension cohort of a phase II trial

BACKGROUND: Cabozantinib is an orally available inhibitor of tyrosine kinases including VEGFR2 and c-MET. We performed a post hoc analysis to find associations between select plasma biomarkers and treatment response in patients (pts) with metastatic castration resistant prostate cancer (mCRPC) who received cabozantinib 100 mg daily as part of a phase 2 non-randomized expansion cohort (NCT00940225). METHODS: Plasma samples were collected at baseline, 6 weeks and at time of maximal response from 81 mCRPC pts with bone metastases, of which 33 also had measurable soft-tissue disease. Levels of 27 biomarkers were measured in duplicate using enzyme-linked immunosorbent assay. Spearman correlation coefficients were calculated for the association between biomarker levels or their change on treatment and either bone scan response (BSR) or soft tissue response according to RECIST. RESULTS: A BSR and RECIST response were seen in 66/81 pts (81 %) and 6/33 pts (18 %) respectively. No significant associations were found between any biomarker at any time point and either type of response. Plasma concentrations of VEGFA, FLT3L, c-MET, AXL, Gas6A, bone-specific alkaline phosphatase, interleukin-8 and the hypoxia markers CA9 and clusterin significantly increased during treatment with cabozantinib irrespective of response. The plasma concentrations of VEGFR2, Trap5b, Angiopoietin-2, TIMP-2 and TIE-2 significantly decreased during treatment with caboznatinib. CONCLUSIONS: Our data did not reveal plasma biomarkers associated with response to cabozantinib. The observed alterations in several biomarkers during treatment with cabozantinib may provide insights on the effects of cabozantinib on tumor cells and on tumor micro-environment and may help point to potential co-targeting approaches

Arrabidaea chica Verlot : controled release formulations with aplications for ulcer lesions

Orientadores: Mary Ann Foglio, João Ernesto de Carvalho, Rodney Alexandre Ferreira RodriguesTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: A espécie Arrabidaea chica, conhecida como crajiru, faz parte da relação nacional de plantas medicinais de interesse ao SUS. Estudos desenvolvidos no CPQBA-UNICAMP comprovaram que o extrato bruto de A. chica possui ação cicatrizante, antiulcerogênica e antioxidante. Este trabalho teve como objetivo delinear e avaliar sistemas farmacêuticos contendo o extrato padronizado de A. chica, para o tratamento de lesões de pele e mucosa. Foram obtidas nanopartículas de quitosana pelo método de reticulação. Dentre os parâmetros avaliados estipulou-se como melhores condições para produção de nanopartículas o emprego do tripolifosfato como reticulante nas proporções de 1:5 (massa) e 1:10 (volume) em relação a quitosana. Para as aplicações tópicas, as nanopartículas de A. chica foram veiculadas em esponjas, filmes absorvíveis de álcool polivinílico ou hidrogéis de ácido hialurônico. Estudos in vitro utilizando fibroblastos humanos demonstraram a atividade das nanopartículas de A. chica na proliferação e migração celular. Estudos in vivo empregando ratos e hamsters forneceram parâmetros para padronização de modelos de mucosite gastrointestinal e oral, respectivamente. A atividade das nanopartículas em úlceras de mucosa foi evidenciada nos modelos de úlcera gástrica induzidas por etanol ou indometacina, com redução das lesões ulcerativas de 76% e 58%, respectivamente, quando comparadas ao grupo controle negativo. O efeito cicatrizante de filmes absorvíveis ou hidrogéis incorporados com as nanopartículas de A. chica foram avaliados em modelo de úlcera dérmica em ratos. A contração da área da ferida chegou a 79% nos animais tratados com os filmes carregados com 0,5 mg de nanopartículas e 85% de contração nos animais que receberam hidrogéis contendo 1,5 mg de nanopartículas. Os resultados demonstraram que a produção de nanopartículas de A. chica e dos sistemas de transporte foi viável, caracterizando-se como uma alternativa válida para a veiculação do extrato de A. chica, além de propiciar a redução da dose necessária para a atividadeAbstract: Arrabidaea chica, known as crajiru, is part of the national Unified Health System from Brazil (SUS) list of medicinal plants of interest. Studies conducted at CPQBA- UNICAMP showed that the species demonstrated wound healing, antiulcerogenic and antioxidant properties. This study aimed the evaluation of pharmaceutical systems employing standardized extract for treatment of skin and mucosal wounds. Chitosan nanoparticles were obtained using the crosslink method. The protocol that showed best conditions to produce nanoparticles, among those tested, used tripolyphosphate as a cross-linker, in 1:5 (mass) and 1:10 (volume) proportions compared to chitosan. For topical applications, A. chica nanoparticles were incorporated into sponges, polyvinyl alcohol absorbable films or hyaluronic acid hydrogels. In vitro studies with human fibroblasts showed the activity of A. chica nanoparticles in cell proliferation and migration. In vivo studies with rats and hamsters provided the standardization parameters for gastrointestinal and mucositis models respectively. A chica nanoparticles demonstrated inhibition of mucosal ulcerations of gastric ulcer induced by ethanol or indomethacin models, improving ulcerative wound healing by 76% and by 58% respectively, in comparison to negative control groups. Those same samples incorporated into absorbable films or hydrogels were evaluated on wound healing activity using a rat dermal ulcer model. Animals treated with loaded films with 0.5 mg of nanoparticles achieved 79% in wound area contraction, whereas animals receiving hydrogels containing 1.5 mg of nanoparticles achieved 85%. These results showed that the production of A. chica nanoparticles and transport systems were practicable, demonstrating to be a valid alternative for incorporation of A. chica extracts, allowing decrease of the required dose to achieve the desired effectDoutoradoFarmacologia, Anestesiologia e TerapeuticaDoutora em Odontologi

Microbiological method for the determination of Amoxicillin potency in oral suspensions

O presente trabalho utiliza o método microbiológico de cilindros em placas para o doseamento da amoxicilina em suspensões orais. Usou-se, como microrganismo-teste, Staphylococcus aureus ATCC 25923 em meio de cultura Ágar Mueller-Hinton. Soluções de amoxicilina com 10, 15, 20, 30, 50 e 100 µg/mL foram empregadas para a curva de calibração. Cilindros foram preenchidos com 50 µL dessas soluções. Os diâmetros dos halos (n=2) foram de 21,5; 23,75; 25,5; 27; 31 e 33,75 milímetros (mm) com coeficientes de variação (CV) de 3,29; 1,49; 2,77; 0; 0 e 1,05%, respectivamente. Analisaram-se três suspensões comerciais de amoxicilina (referência, similar e genérico) com potência declarada de 250 mg/5mL. Soluções de 20 µg/mL, preparadas com as suspensões, apresentaram halos de inibição com médias (n=5) de 25,33; 25,33 e 25,17 mm. As concentrações determinadas foram de 251,6; 251,6 e 244,74 mg/5mL, o que corresponde a 100,6; 100,6 e 97,74% da potência declarada, com CV de 2,28; 2,28 e 1,15%, respectivamente. Concluiu-se que as amostras analisadas estavam dentro dos limites preconizados pela Farmacopéia Brasileira (90 a 120% da potência declarada). A partir dos resultados obtidos, observou-se que o método microbiológico de cilindros em placas é adequado e válido para o doseamento da potência da amoxicilina em suspensões orais.The present work reports a microbiological assay, applying the cylinder-plate method, for the determination of amoxicillin in oral suspension. Stains of Staphylococcus aureus ATCC 25923 were used as a test microorganism in Mueller-Hinton Agar. Amoxicillin solutions of 10, 15, 20, 30, 50 and 100 µg/mL were used for the calibration curve. The cylinders were fulfilled with 50 µL of these solutions. The halo diameters (n=2) were 21.5; 23.75; 25.5; 27; 31 and 33.75 mm and the coefficients of variation (CV) were 3.29; 1.49; 2.77; 0; 0 and 1.05%, respectively. Three commercial amoxicillin suspensions (reference, similar and generic) with declared potency of 250 mg/5mL were studied through the method above. Solutions of 20 µg/mL were then prepared and showed inhibition halos media (n=5) of 25.33; 25.33 and 25.17 mm. The concentrations were 251.6; 251.6 and 244.74 mg/5mL which correspond to 100.6; 100.6 and 97.74% of the declared potency, with CV of 2.28; 2.28 and 1.15%, respectively. It was concluded that the commercial samples used in this assay were situated on the limits indicated by the Brazilian Pharmacopoeia (90 to 120% of the declared potency). The results pointed that the microbiological assay, applying the cylinder-plate method, can be used for the determination of amoxicillin potency in oral suspensions.Colegio de Farmacéuticos de la Provincia de Buenos Aire