Solid Lipid Nanoparticles Based on Babassu Oil and Copaiba Oleoresin: A Promising Approach for Prostate Cancer Therapy.

Solid lipid nanoparticles (SLNs) represent promising nanostructures for drug delivery systems. This study successfully synthesized SLNs containing different proportions of babassu oil (BBS) and copaiba oleoresin (COPA) via the emulsification-ultrasonication method. Before SLN synthesis, the identification and quantification of methyl esters, such as lauric acid and ß-caryophyllene, were performed via GC-MS analysis. These methyl esters were used as chemical markers and assisted in encapsulation efficiency experiments. A 22 factorial design with a center point was employed to assess the impact of stearic acid and Tween 80 on particle hydrodynamic diameter (HD) and polydispersity index (PDI). Additionally, the effects of temperature (8 ± 0.5 °C and 25 ± 1.0 °C) and time (0, 7, 15, 30, 40, and 60 days) on HD and PDI values were investigated. Zeta potential (ZP) measurements were utilized to evaluate nanoparticle stability, while transmission electron microscopy provided insights into the morphology and nanometric dimensions of the SLNs. The in vitro cytotoxic activity of the SLNs (10 µg/mL, 30 µg/mL, 40 µg/mL, and 80 µg/mL) was evaluated using the MTT assay with PC-3 and DU-145 prostate cancer cell lines. Results demonstrated that SLNs containing BBS and COPA in a 1:1 ratio exhibited a promising cytotoxic effect against prostate cancer cells, with a percentage of viable cells of 68.5% for PC-3 at a concentration of 30 µg/mL and 48% for DU-145 at a concentration of 80 µg/mL. These findings underscore the potential therapeutic applications of SLNs loaded with BBS and COPA for prostate cancer treatment.

Evaluation of Silybin Nanoparticles against Liver Damage in Murine Schistosomiasis mansoni Infection.

Silybin (SIB) is a hepatoprotective drug known for its poor oral bioavailability, attributed to its classification as a class IV drug with significant metabolism during the first-pass effect. This study explored the potential of solid lipid nanoparticles with (SLN-SIB-U) or without (SLN-SIB) ursodeoxycholic acid and polymeric nanoparticles (PN-SIB) as delivery systems for SIB. The efficacy of these nanosystems was assessed through in vitro studies using the GRX and Caco-2 cell lines for permeability and proliferation assays, respectively, as well as in vivo experiments employing a murine model of Schistosomiasis mansoni infection in BALB/c mice. The mean diameter and encapsulation efficiency of the nanosystems were as follows: SLN-SIB (252.8 ± 4.4 nm, 90.28 ± 2.2%), SLN-SIB-U (252.9 ± 14.4 nm, 77.05 ± 2.8%), and PN-SIB (241.8 ± 4.1 nm, 98.0 ± 0.2%). In the proliferation assay with the GRX cell line, SLN-SIB and SLN-SIB-U exhibited inhibitory effects of 43.09 ± 5.74% and 38.78 ± 3.78%, respectively, compared to PN-SIB, which showed no inhibitory effect. Moreover, SLN-SIB-U demonstrated a greater apparent permeability coefficient (25.82 ± 2.2) than PN-SIB (20.76 ± 0.1), which was twice as high as that of SLN-SIB (11.32 ± 4.6) and pure SIB (11.28 ± 0.2). These findings suggest that solid lipid nanosystems hold promise for further in vivo investigations. In the murine model of acute-phase Schistosomiasis mansoni infection, both SLN-SIB and SLN-SIB-U displayed hepatoprotective effects, as evidenced by lower alanine amino transferase values (22.89 ± 1.6 and 23.93 ± 2.4 U/L, respectively) than those in control groups I (29.55 ± 0.7 U/L) and I+SIB (34.29 ± 0.3 U/L). Among the prepared nanosystems, SLN-SIB-U emerges as a promising candidate for enhancing the pharmacokinetic properties of SIB.

In vitro and in situ caries-preventive effect of a new combined fluoride and calcium experimental nanocomposite solution

Abstract To assess the in vitro and in situ effect of experimental combined fluoride and calcium nanocomposite solutions on dental caries prevention. Nanocompound mesoporous silica (MS) with calcium (Ca) and sodium fluoride (NaF) - (MSCaNaF); MS with NaF (MSNaF), NaF solution (positive control), and deionized water (negative control - CG) were studied. The specimens (n=130) were submitted in vitro to a multispecies biofilm in the presence of 2% sucrose. After 24 h and 48 h, the culture medium pH, the percent of surface mineral loss (%SML), and lesion depth (ΔZ) were analyzed. In the in situ study, 10 volunteers participated in four phases of 7-days each. The products were applied on the specimens (n=240) before 20% sucrose solution drips. The polysaccharides (SEPS and IEPS), %SML and roughness (Sa) were evaluated. There was an in vitro decrease in pH values in 24h and 48h, compared to baseline. The MSCaNaF and MSNaF groups obtained lower values of %SML and ΔZ (p < 0.05) than CG and NaF after 24h and were similar to NaF after 48h (p<0.05). In situ results showed similar SEPS and IEPS among all groups after 48h. An after 7-days, the nanocomposites had similar values (p>0.05), while NaF was similar to CG (p>0.05). After 48h, the MSCaNaF and MSNaF reduced the %SML (p<0.05). After 7-days, both experimental nanocomposites were similar to NaF (p>0.05). Regarding Sa, MSCaNaF was better than NaF for both periods (p<0.05). The nanocomposites controlled the in vitro and in situ enamel demineralization, mainly in the initial periods.

Copaiba Oil Resin Exerts an Additive Effect to Babassu Oil on Behavioral Changes in Human Endometriotic Cell Cultures.

BACKGROUND: Current drugs for the treatment of endometriosis are not able to completely cure the condition, and significant side effects hinder the continuation of treatment. Therefore, it is necessary to search for new drug candidates. In the present paper, the use of plant extracts is highlighted. Babassu oil and Copaiba oil resin have several therapeutic properties. We investigated the in vitro effects of two nanoemulsions containing oil extracted from Babassu (Orbignya speciosa) nuts (called SNEDDS-18) and/or oil resin extracted from Copaiba trunk (Copaifera langsdorffii) (called SNEDDS-18/COPA) on cultured human eutopic endometrium stromal cells from endometrial biopsies of patients without (CESC) and with (EuESC) endometriosis as well as human stromal cells from biopsies of endometriotic lesions (EctESC). METHODS: CESC, EuESC, and EctESC were taken and treated with SNEDDS-18 and SNEDDS-18/COPA to evaluate their effects on cytotoxicity, cell morphology, proliferation, and signaling pathways. RESULTS: After 48 h of incubation with SNEDDS-18 and SNEDDS-18/COPA, cell viability and proliferation were inhibited, especially in EctESC. The lowest concentration of both nanoemulsions reduced cell viability and proliferation and broke down the cytoskeleton in EctESCs. After 24 h of treatment a decrease in IL-1, TNF-α, and MCP-1 was observed, as well as an increase in IL-10 production. CONCLUSIONS: Both nanoemulsions can affect endometriotic stromal cell behaviors, thus revealing two potential candidates for new phytotherapeutic agents for the management of endometriosis.

In vitro effect of experimental nanocomposites solutions on the prevention of dental caries around orthodontic brackets

Abstract This study aimed to evaluate the in vitro effect of a single application of experimental nanocomposite solutions on the prevention of dental caries around orthodontic brackets. The specimens were exposed to mesoporous silica (MS) nanocomposites containing fluoride by association with titanium tetrafluoride (TiF4) or sodium fluoride (NaF). Nanocomposites also could contain calcium and groups were described as MSCaTiF4, MSTiF4, MSCaNaF, MSNaF, and controls (TiF4, and NaF). Specimens were subjected to the formation of a multispecies biofilm to generate a cariogenic challenge. After 24h, both pH and total soluble fluoride concentration of the culture medium were assessed. Mineral loss was evaluated by percentage of surface mineral loss (%SML), mineral volume variation (ΔZ) of inner enamel and polarized light microscopy (PL). Linear (Ra) and volumetric (Sa) surface roughness and scanning electronic microscopy (SEM) were used to assess enamel topography. Statistical analyses were conducted considering p<0.05. MSNaF had the highest value of culture medium pH after cariogenic challenge, similarly to MSTiF4. All nanocomposite solutions released less fluoride than their controls NaF and TiF4 (p<0.05). All nanocomposite solutions presented lower %SML compared to their respective control groups (p<0.05). Lower Ra, Sa and ΔZ were observed for experimental groups compared to TiF4 (p<0.05). The results were confirmed by PL and SEM analysis. The experimental nanocomposite solutions contributed for lower enamel demineralization around orthodontic brackets.

RESUMO Este estudo teve como objetivo avaliar o efeito in vitro de uma única aplicação de soluções experimentais de nanocompósitos na prevenção de cárie dentária em braquetes ortodônticos. Os espécimes foram expostos a nanocompósitos de sílica mesoporosa (MS) contendo fluoreto por associação com tetrafluoreto de titânio (TiF4) ou fluoreto de sódio (NaF). Os nanocompósitos também podem conter cálcio e os grupos foram descritos como MSCaTiF4, MSTiF4, MSCaNaF, MSNaF e controles (TiF4 e NaF). Os espécimes foram submetidos à formação de um biofilme multiespécie para gerar um desafio cariogênico. Após 24h, o pH e a concentração de flúor solúvel total do meio de cultura foram avaliados. A perda mineral foi avaliada pela porcentagem de perda mineral superficial (% SML), variação do volume mineral (ΔZ) do esmalte interno e microscopia de luz polarizada (PL). A rugosidade superficial linear (Ra) e volumétrica (Sa) e a microscopia eletrônica de varredura (MEV) foram utilizadas para avaliar a topografia do esmalte. As análises estatísticas foram realizadas considerando p <0,05. MSNaF apresentou o maior valor de pH do meio de cultura após o desafio cariogênico, semelhante ao MSTiF4. Todas as soluções de nanocompósitos liberaram menos flúor do que seus controles NaF e TiF4 (p <0,05). Todas as soluções de nanocompósitos apresentaram% SML menor em comparação com seus respectivos grupos de controle (p <0,05). Ra, Sa e ΔZ menores foram observados para os grupos experimentais em comparação ao TiF4 (p <0,05). Os resultados foram confirmados por análises PL e SEM. As soluções experimentais de nanocompósitos contribuíram para a menor desmineralização do esmalte ao redor dos braquetes ortodônticos.

Comparative effects of cpp-acp and xylitol f-varnishes on the reduction of tooth erosion and its progression

Abstract Evaluated the effect of CPP-ACP/NaF and xylitol/NaF varnishes in reduce erosion and progression of erosion. Forty enamel blocks were divided into four groups (n=10): G1=CPP-ACP/NaF varnish (MI varnishTM); G2=xylitol/NaF varnish (Profluorid®); G3=NaF varnish (Duraphat®, positive control) and G4=deionized water (MilliQ®, negative control). Samples were immersed in Sprite ZeroTM (pH 2.58, 4x/day, 3 days), in between immersions, the specimens stayed in artificial saliva. After 3 days of erosion, the eroded area was divided in two (half of one received an additional varnish layer while the other half repeated the same 3-day erosion cycle). The 3D, non-contact profilometry technique was used to determinate tooth structure loss (TSL) and surface roughness (SR). Scanning electron microscopy (SEM) and 3D images were utilized to evaluate the topography of the samples. Mann-Whitney, one-way ANOVA and Tukey tests were used (significance level of 0.05%). SEM and 3D images were descriptively evaluated. After 3 or 6 days of erosion, all tested varnishes were better than G4 (p<0.05) for TSL and SR. In addition, G1 had lower values for TSL than G3 (p<0.05) after 3 days of erosion. Under SEM and 3D images observation, all groups presented porosity, irregularities and depressions on the surface enamel after 3 and 6 days of erosion, more pronounced in G4. An application of topical NaF varnishes was effective in reducing TSL and enamel roughness after erosion challenges, being the CCP-ACP/NaF varnish more effective than NaF varnish and water after 3 days of erosion.

Resumo Avaliou-se o efeito dos vernizes CPP-ACP/NaF e xilitol/NaF na redução da erosão e progressão da erosão. Quarenta blocos de esmalte foram divididos em quatro grupos (n=10): G1=verniz CPP-ACP/NaF (verniz MITM); G2=verniz xilitol/NaF (Profluorid®); G3=verniz NaF (Duraphat®, controle positivo) e G4=água desionizada (MilliQ®, controle negativo). As amostras foram imersas em refrigerante Sprite ZeroTM (pH 2.58, 4x/dia, 3 dias), entre imersões, os espécimes ficaram em saliva artificial. Após 3 dias de erosão, a área erodida foi dividida em duas (metade recebeu uma camada adicional de verniz, enquanto a outra metade repetiu o mesmo ciclo de erosão de 3 dias). A técnica de perfilometria 3D de não contato foi utilizada para determinar a perda de estrutura dentária (PED) e rugosidade superficial (RS). Microscopia eletrônica de varredura (MEV) e imagens em 3D foram utilizadas para avaliar a topografia das amostras. Testes de Mann-Whitney, One-way ANOVA e Tukey foram utilizados (nível de significância de 0,05%). Imagens do MEV e 3D foram avaliadas descritivamente. Após 3 ou 6 dias de erosão, todos os vernizes testados foram melhores que G4 (p<0,05) para PED e RS. Além disso, o G1 apresentou menores valores de PED do que o G3 (p<0,05) após 3 dias de erosão. Observando as imagens em MEV e 3D, todos os grupos apresentaram porosidade, irregularidades e depressões no esmalte superficial após 3 e 6 dias de erosão, sendo mais pronunciados no G4. Uma aplicação tópica de vernizes fluoretados foi eficaz na redução da rugosidade e PED do esmalte após desafios de erosão. Além disso, o grupo CPP-ACP/NaF teve melhor desempenho na redução da PED quando comparado ao verniz de NaF e a água, após 3 dias de erosão.

Nanoparticles Loaded with a New Thiourea Derivative: Development and In vitro Evaluation Against Leishmania amazonensis.

BACKGROUND: Leishmaniasis is a neglected tropical disease caused by protozoa of the genus Leishmania. Current treatments are restricted to a small number of drugs that display both severe side effects and a potential for parasites to develop resistance. A new N-(3,4-methylenedioxyphenyl)-N'- (2-phenethyl) thiourea compound (thiourea 1) has shown promising in vitro activity against Leishmania amazonensis with an IC50 of 54.14 µM for promastigotes and an IC50 of 70 µM for amastigotes. OBJECTIVE: To develop a formulation of thiourea 1 as an oral treatment for leishmaniasis, it was incorporated into Nanoparticles (NPs), a proven approach to provide long-acting drug delivery systems. METHODS: Poly (D,L-Lactic-co-Glycolic Acid) (PLGA) polymeric NPs containing thiourea 1 were obtained through a nanoprecipitation methodology associated with solvent evaporation. The NPs containing thiourea 1 were characterized for Encapsulation Efficiency (EE%), reaction yield (% w/w), surface charge, particle size and morphology by Transmission Electron Microscopy (TEM). RESULTS: NPs with thiourea 1 showed an improved in vitro leishmanicidal activity with a reduction in its cytotoxicity against macrophages (CC50>100 µg/mL) while preserving its IC50 against intracellular amastigotes (1.46 ± 0.09 µg/mL). This represents a parasite Selectivity Index (SI) of 68.49, which is a marked advancement from the reference drug pentamidine (SI = 30.14). CONCLUSION: The results suggest that the incorporation into NPs potentiated the therapeutic effect of thiourea 1, most likely by improving the selective delivery of the drug to the phagocytic cells that are targeted for infection by L. amazonensis. This work reinforces the importance of nanotechnology in the acquisition of new therapeutic alternatives for oral treatments.

The use of a new calcium mesoporous silica nanoparticle versus calcium and/or fluoride products in reducing the progression of dental erosion

Abstract Objective There is increasingly common the consumption more times a day of foods and acidic drinks in the diet of the population. The present study aimed to evaluate and compare the effects of a calcium mesoporous silica nanoparticle single application of other calcium and/or fluoride products in reducing the progression of dental erosion. Methodology Half of the eroded area was covered of 60 blocks of enamel, after which the block was submitted to the following treatments: (Ca2+-MSN), casein phosphopeptide-amorphous calcium phosphate (CPP-ACP); CPP-ACP/F-(900 ppm F−); titanium tetrafluoride (TiF4 1%) (positive control); sodium fluoride (NaF 1.36%) (positive control); and Milli-Q® water (negative control) before being submitted to a second erosive challenge. A surface analysis was performed via a three-dimensional (3D) noncontact optical profilometry to assess the volumetric roughness (Sa) and tooth structure loss (TSL) and and through scanning electron microscopy (MEV). An analysis of variance (ANOVA) and Tukey's test were performed. Results Regarding Sa, all experimental groups exhibited less roughness than the control (p<0.05). The TSL analysis revealed that the Ca2+-MSN and NaF groups were similar (p>0.05) and more effective in minimizing tooth loss compared with the other groups (p<0.05). Conclusions The Ca2+-MSN and NaF treatments were superior compared with the others and the negative control.

Comparative Effect of Calcium Mesoporous Silica Versus Calcium and/or Fluoride Products on the Reduction of Erosive Tooth Wear and Abrasive Enamel Lesion

Abstract Objective: To evaluate the effect of a calcium nanocompound on the reduction of erosive tooth wear and abrasion. Material and Methods: Bovine enamel specimens (BE), were randomly assigned to the following groups (n = 10): G1 = Calcium mesoporous silica nanoparticles (Ca2+MSNs); G2 = casein phosphopeptide-amorphous calcium phosphate (CPP-ACP, 2% CPP-ACP, GC®); G3 = casein phosphopeptide-amorphous calcium fluoride phosphate (CPP-ACFP, 2% CPP-ACP + 900 ppm F-, GC®); G4 = sodium fluoride NaF (900 ppm F-, positive control); and G5 = distilled and deionized water (negative control). Each product was applied to the exposed area for one minute, three times per day for three consecutive days, and followed by the immersion of the specimens in Sprite Zero™ - a low-pH solution (2.58) for five minutes (Coca-Cola™). After the first and last erosive challenges of the day, the specimens were submitted to abrasion in a toothbrush machine for 15 seconds (200 g/BE). The specimens were analysed using 3D non-contact optical profilometry, with tooth structure loss (TSL) measurements and scanning electron microscopy (SEM). TSL values were analysed by Kruskal-Wallis and Mann-Whitney tests (p<0.05). Results: There were no significant differences between G1 (10.95 µm) and G3 (10.80 µm) treatments for TSL values; however both resulted in significantly reduced TSL values compared with the G5 (16.00 µm) (p<0.05). The G4 (12.26 µm) showed no statistically significant difference when compared to the G5 (16.00 µm). The groups G1 and G3 presented higher surface preservation than the G5. Conclusion: Ca2+MSNs was effective for reducing tooth surface loss caused by erosive tooth wear and abrasion.

Translational impact of novel widely pharmacological characterized mofezolac-derived COX-1 inhibitors combined with bortezomib on human multiple myeloma cell lines viability.

A set of novel diarylisoxazoles has been projected using mofezolac (1) as a lead compound to investigate structure-inhibitory activity relationships of new compounds and the cyclooxygenases (COXs) catalytic activity. Mofezolac was chosen because is the most potent and selective reversible COX-1 inhibitor [COX-1 IC50 = 0.0079 µM and COX-2 IC50 > 50 µM, with a selectivity index (SI) in favor of COX-1 higher than 6300]. Seventeen new compounds were synthesized in fair to good yields and evaluated for their COXs inhibitory activity and selectivity. SIs ranged between 1 and higher than 1190.3,4-Bis(4-methoxyphenyl)-5-vinylisoxazole (22) has the highest SI with COX-1 IC50 = 0.042 µM and COX-2 IC50 > 50 µM. 1 and 22 were superior to aspirin in inhibiting platelet aggregation (IC50 = 0.45, 0.63 and 1.11 µM, respectively) in human platelet rich plasma (hPRP) assay. They did not induce blood coagulation and hemolysis, and are neither genotoxic nor mutagen. 1 and 22 slightly increase bortezomib cytotoxic effect on multiple myeloma (MM) cell lines (NCI-H929 and RPMI-8226) and affects MM cell cycle and apoptosis when co-administered with the proteasome inhibitor bortezomib, a drug clinically used to treat plasma cell neoplasms including MM. In addition, structure-based binding mode of 1 and 22, through Fingerprints for Ligands and Proteins (FLAG) calculation, allowed to explain the one order of magnitude difference between COX-1 IC50 values of the two compounds. Specifically, the higher inhibitory potency seems due to the formation of a H-bond between COX-1 S530 and the carboxyl, present in 1 and absent in 22.

Development of a new formulation of roflumilast for pulmonary drug delivery to treat inflammatory lung conditions.

The aim of this study was to develop roflumilast dry powder inhaler (DPI) formulations by spray drying using hydroxypropyl-ß-cyclodextrin (HPßCD) and to determine their suitability for pulmonary delivery. Different feed solution concentrations, solvent systems and spray drying parameters were used to obtain the formulations which were characterized using X-ray powder diffraction, thermal analysis, scanning electron microscopy, particle size distribution, bulk and tapped density, specific surface area, dynamic vapour sorption, in vitro deposition properties using a Next Generation Impactor (NGI) and transepithelial permeability. Microparticles spray dried from ethanol were wrinkled and amorphous, exhibiting high glass transition temperatures while those from methanol:n-butyl acetate consisted of irregularly shaped porous particles partially crystalline. All formulations presented low density, particle size and residual solvent content exhibiting high depositon in the lower stages of the NGI. Mass median aerodynamic diameters (MMADs) were in the range of 3.32-4.49 µm, with high fine particle fractions (FPF < 5 µm). Stability studies demonstrated no significant modifications in the solid-state nature and in the aerolisation performance of the selected formulation which presented a Papp of 8.73 × 10-6 ±â€¯4.70 × 10-7 cm/s. The developed roflumilast DPI formulations have potential therapeutic applications in the treatment of lung diseases.

Synthesis and evaluation of the cytotoxic activity of Furanaphthoquinones tethered to 1H-1,2,3-triazoles in Caco-2, Calu-3, MDA-MB231 cells.

Naphthoquinones and 1,2,3-triazoles are structural pharmacophore that is known to impart several cancer cells. This work shows a synthetic methodology to obtain hybrid molecules involving naphthoquinone and triazol scaffold as multiple ligands. A simple and efficient synthetic route was used to prepare a series of sixteen compounds being eight 2-(1-aryl-1H-1,2,3-triazol-4-yl)-2,3-dihydronaphtho[1,2 b]furan-4,5-diones and eight 2-(1-aryl-1H-1,2,3-triazol-4-yl)-2,3-dihydronaphtho[2,3-b]furan-4,9-diones. These compounds were tested in MDA-MB231, Caco-2 and Calu-3 human cancer cells, and among them 7a was the most selective compound on Caco-2 cells, the most sensitized cell line in this study. In silico study suggest that the blockage of topoisomerase I and IIα may be one of the mechanisms of action responsible for the cytotoxic effect of 7a in Caco-2 cells.

Development and characterization of poly(lactic-co-glycolic) acid nanoparticles loaded with copaiba oleoresin.

Copaiba oleoresin (CPO), obtained from Copaifera landgroffii, is described as active to a large number of diseases and more recently in the endometriosis treatment. In this work, poly(lactic-co-glycolic acid) (PLGA) nanoparticles containing CPO were obtained using the design of experiments (DOE) as a tool to optimize the production process. The nanoparticles optimized by means of DOE presented an activity in relation to the cellular viability of endometrial cells. The DOE showed that higher amounts of CPO combined with higher surfactant concentrations resulted in better encapsulation efficiency and size distribution along with good stability after freeze drying. The encapsulation efficiency was over 80% for all produced nanoparticles, which also presented sizes below 300 nm and spherical shape. A decrease in viability of endometrial stromal cells from ectopic endometrium of patients with endometriosis and from eutopic endometriotic lesions was demonstrated after 48 h of incubation with the CPO nanoparticles. The nanoparticles without CPO were not able to alter the cell viability of the same cells, indicating that this material was not cytotoxic to the tested cells and suggesting that the effect was specific to CPO. The results indicate that the use of CPO nanoparticles may represent a promising alternative for the treatment of endometriosis.

Development and validation of a stability-indicating HPLC method for topiramate using a mixed-mode column and charged aerosol detector.

The analysis of topiramate in the presence of its main degradation products is challenging due to the absence of chromophore moieties and their wide range of polarity. Mixed-mode chromatography has been used in such cases because it combines two or more modes of separation. Charged aerosol detector is also an alternative since its detection is independent of optical properties and analyte ionization. This study is aimed to develop and validate two new stability-indicating methods by high-performance liquid chromatography for the main degradation products of topiramate using mixed-mode chromatography and a charged aerosol detector. Method 1 employed an Acclaim Trinity P1® column (3.0 mm × 150 mm, 2.7 µm) with a mobile phase comprising of 80% ammonium acetate buffer (20 mM, pH 4.0) and 20% methanol at a flow rate of 0.5 mL/min at 35°C. Method 2 utilized a C18 Acclaim 120® column (4.6 mm × 250 mm; 5 µm) with ACN/water (50:50) at a flow rate of 0.6 mL/min at 50°C. Validation of the two methods demonstrated excellent performance with respect to linearity, precision, accuracy, and selectivity. The limits of detection for topiramate, fructose, sulfate, sulfamate, and compound A were 2.97, 12.08, 4.02, 13.91, and 3.94 µg/mL, respectively.

Fluoride Varnishes against Dental Erosion Caused by Soft Drink Combined with Pediatric Liquid Medicine

Abstract The present study evaluated the effect of NaF and CPP-ACP/NaF varnishes to reduce erosion produced by soft drink (SD) combined or not with pediatric liquid medicine. Enamel specimens were pre-treated with fluoride varnish, according to the following groups: NaF varnish (Duraphat®) or CPP-ACP/NaF varnish (MI varnishTM). Two types of erosive cycles were made: by soft drink erosion (SDE) or by pediatric liquid medicine plus soft drink erosion (PLM/SDE). Bovine enamel specimens were randomly assigned in six groups (n=10): G1=NaF + SDE; G2=CPP-ACP/NaF + SDE; G3=Distilled and deionized (DD) water + SDE; G4=NaF + PLM/SDE; G5=CPP-ACP/NaF + PLM/SDE and G6=DD water + PLM/SDE. Before treatments, the sample surface was divided in two areas (unexposed area-UA and exposed area-EA). The specimens were evaluated by 3D non-contact profilometry technique to determinate tooth structure loss (TSL) and surface roughness (Sa). Scanning electron microscopy (SEM) analysis was also performed. After SDE, G2 presented the lowest TSL values compared to G3 (p=0.008). G1 and G2 did not differ between them (p=0.203) and no groups differed among them despite Sa. Regarding TSL and Sa, G4 and G5 differed from G6 (p=0.0001), but not between them (p=1.00). Examining 3D and SEM images, the greatest differences between UA and EA were observed for G3 and G6. CPP-ACP/NaF varnish seems to be a promising treatment to reduce enamel loss from the erosion produced by a soft drink. Both varnishes also showed capacity to reduce TSL and Sa after erosion by soft drink combined to pediatric liquid medicine.

Resumo O presente estudo avaliou o efeito dos vernizes de NaF e CPP-ACP/NaF na redução da erosão promovida por refrigerante e associada a um medicamento líquido pediátrico. Os espécimes de esmalte foram pré-tratados com verniz fluoretado, de acordo com o grupo de alocação: verniz NaF (Duraphat®) ou verniz CPP-ACP NaF (verniz MITM). Dois tipos distintos de desafio erosivo foram realizados: erosão com refrigerante (ER) ou erosão com medicamento líquido pediátrico e refrigerante (MLP/ER). Espécies de esmalte bovino foram aleatorizados em seis grupos (n=10): G1 = NaF + ER; G2 = CPP-ACP/NaF + ER; G3 = Água destilada e deionizada (DD) + ER; G4 = NaF + MLP/ER; G5 = CPP-ACP/NaF + MLP/ER e G6 = DD água + MLP/ER. Antes dos tratamentos, a superfície das amostras foi dividida em duas áreas (não exposta-NE e área exposta-AE). Os espécimes foram avaliados pela técnica de perfilometria 3D de não-contato para determinar a perda de estrutura dentária (PED) e a rugosidade superficial (RS). A microscopia eletrônica de varredura (MEV) também foi utilizada. Após ER, G2 apresentou os menores valores de PED comparado ao G3 (p=0,008). G1 e G2 não diferiram entre si (p=0,203) e não houve diferença entre os grupos no que diz respeito a RS. Os resultados de PED e RS para a MLP/ER mostraram que G4 e G5 diferiram de G6 (p=0,0001), mas não diferiram entre si (p=1,00). Examinando as imagens 3D da perfilometria e de MEV, as maiores diferenças entre UA e EA foram observadas para G3 e G6. O verniz CPP-ACP/NaF parece ser um tratamento promissor para reduzir a perda de esmalte por erosão produzida por refrigerante e ambos os vernizes mostraram capacidade em reduzir a PED e RS após erosão com medicamento líquido pediátrico associado a refrigerante.

Targeted Nanosystems to Prostate Cancer.

Prostate cancer remains an increasingly common malignancy worldwide. Many advances in drug development have been achieved for the conventional treatments; however, chemotherapeutic agents are distributed nonspecifically in the body where they affect both prostate cancer and healthy cells. Limited dose achievable within the prostate tumor and suboptimal treatment due to excessive toxicities reveal the importance of the development of more specific mechanisms and ways of drug targeting to prostate tumor. In this context, nanotechnology, molecular biology and biochemistry have been applied in the pharmaceutical area for development of new targeted drug delivery nanosystems in order to improve its pharmacokinetic profile, raise the effectiveness of treatment; reduce side effects due to the preferential accumulation in prostate cancer cells, causing low concentrations in healthy tissues; and/or increase the drug chemical stability for improving the prostate cancer therapeutic. Thus, in this review, we will discuss the molecular and biochemical basis of prostate cancer as well as the advantages and disadvantages of conventional clinical treatments, different types and basic characteristics of nanosystems; how these systems can be targeted to prostate cancer, show successful patent examples of prostate cancer targeted nanosystems and present perspectives for the next 10-20 years in this area.

Development and pharmacological evaluation of in vitro nanocarriers composed of lamellar silicates containing copaiba oil-resin for treatment of endometriosis.

In this work, newly developed nanocomposites based upon lamellar silicates are evaluated to determine their potential in controlling endometriosis. The preparation of the new nanocarriers is detailed, properties characterized and in vitro pharmacological evaluation performed. The nanocomposites in this study were obtained from the reaction of copaiba oil-resin (COPA) with the polymer polyvinylpyrrolidone (PVP K-30). COPA was selected due to its antiinflammatory and anticancer activities along with the organophilic derivatives of sodium montmorillonite, Viscogel B8, S7 and S4. The results indicated that it was feasible to obtain a good yield of a COPA nanocomposite using a simple process. Intercalation was confirmed by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC). In vitro release experiments demonstrated that COPA was released from the nanocomposite in a delayed fashion. Whereas, in vitro pharmacological studies showed a reduction in viability and proliferation of endometriotic cell cultures upon COPA nanocomposite treatment, suggesting that the system developed here can be a promising alternative therapy for the oral treatment of endometriosis.

The oil-resin of the tropical rainforest tree Copaifera langsdorffii reduces cell viability, changes cell morphology and induces cell death in human endometriotic stromal cultures.

OBJECTIVES: The hormonal treatment for endometriosis frequently fails to completely eradicate endometriotic implants. A new therapeutic treatment is needed. This study investigates the in-vitro effect of Copaifera langsdorffii oil-resin on human eutopic and ectopic endometrium stromal cell cultures (EuESCs and EctESCs). METHODS: A nanocomposite system containing the copaiba oil-resin (NanoCOR) was developed and acute toxicity test was performed. Endometrial stromal cells (ESCs) from non-endometriotics controls (CESCs), EuESCs and EctESCs were isolated and treated with different concentrations of NanoCOR, at different time intervals to evaluate its effect on cell morphology, proliferation, viability, necrosis and apoptosis induction. KEY FINDINGS: When treated with 50 µg/ml of NanoCOR, the morphology of EctESCs changed, as the actin microfilaments were disorganized, disassembled or disrupted. Moreover, at 24 h of treatment with NanoCOR, the EctESCs viability was inhibited, and a significant number of these cells underwent apoptosis. In EuESCs, these effects were observed only at 48 h. Finally, the treatment of EctESCs with NanoCOR increased the lactate dehydrogenase release into the extracellular medium more than in EuESCs. CONCLUSIONS: Our data indicate that NanoCOR has a greater impact on the behaviour of human endometriotic stromal cells than on the eutopic endometrium stromal cells, supporting the idea that NanoCOR should be further investigated as a novel and valuable alternative to treat endometriosis.

Human thromboxane synthase: comparative modeling and docking evaluation with the competitive inhibitors Dazoxiben and Ozagrel.

Thromboxane synthase (TXAS) is a P450 epoxygenase that synthesizes thromboxane A2 (TXA2), a potent mediator of platelet aggregation, vasoconstriction and bronchoconstriction. This enzyme plays an important role in several human diseases, including myocardial infarction, stroke, septic shock, asthma and cancer. Despite of the increasing interest on developing TXAS inhibitors, the structure and activity of TXAS are still not totally elucidated. In this study, we used a comparative molecular modeling approach to construct a reliable model of TXAS and analyze its interactions with Dazoxiben and Ozagrel, two competitive inhibitors. Our results were compatible with experimental published data, showing feasible cation-π interaction between the iron atom of the heme group of TXAS and the basic nitrogen atom of the imidazolyl group of those inhibitors. In the absence of the experimental structure of thromboxane synthase, this freely available model may be useful for designing new antiplatelet drugs for diseases related with TXA2.

Assessment of analytical techniques for characterization of crystalline clopidogrel forms in patent applications

The aim of this study was to evaluate two important aspects of patent applications of crystalline forms of drugs: (i) the physicochemical characterization of the crystalline forms; and (ii) the procedure for preparing crystals of the blockbuster drug clopidogrel. To this end, searches were conducted using online patent databases. The results showed that: (i) the majority of patent applications for clopidogrel crystalline forms failed to comply with proposed Brazilian Patent Office guidelines. This was primarily due to insufficient number of analytical techniques evaluating the crystalline phase. In addition, some patent applications lacked assessment of chemical/crystallography purity; (ii) use of more than two analytical techniques is important; and (iii) the crystallization procedure for clopidogrel bisulfate form II were irreproducible based on the procedure given in the patent application.

Este trabalho tem como objetivo avaliar dois aspectos importantes em um pedido de patente de formas cristalinas de fármacos: (i) caracterização físico-química das formas cristalinas e (ii)o procedimento de preparo da forma II do fármaco clopidogrel, um blockbuster de vendas. Realizaram-se buscas em bancos de dados patentários on line. Os resultados mostraram que (i) a maioria dos pedidos de patente de formas cristalinas do clopidogrel não se adequam com proposta do INPI devido ao número insuficiente de técnicas analíticas utilizadas na caracterização da fase cristalina. Ainda, em alguns pedidos de patente não há a presença da avaliação da pureza química/cristalográfica; (ii) a importância de se utilizar mais de duas técnicas de avaliação e (iii) que não foi possível a reprodução da cristalização com o procedimento apresentado no pedido de patente.