RESUMO
New therapeutics such as antisense oligonucleotides, small interfering RNA and peptide-drug conjugates are taking great relevance in the pharmaceutical industry due to their specificity of action and their improved safety profile. However, they could present bioavailability issues due to their hydrophilic nature, such as BCS class III drugs. Therefore, the formation of ion pairs of these type of molecules allows modifying their physicochemical characteristics such as polarity and lipophilicity leading to improved permeability. By carrying out a tailored synthesis, it is possible to obtain complexes with greater stability and better performance in vitro and in vivo, where their correlation with physicochemical properties continues to be a growing field of research. Moreover, ionic liquids (IL), which are substances that melt below 100 °C, have enabled modifying various drug properties, showing promising results in vitro-in vivo, especially when they are included in suitable drug delivery systems, such as nanoparticles, microparticles, self-emulsifying drug delivery systems, and transdermal patches, among others. The drug-IL is formed from the therapeutic agent and a counterion, mainly by ionic interactions, and resulting in a wide variety of derivatives with different properties. However, the pharmaceutical field is limited to the use of some excipients or GRAS (generally recognized as safe) substances, so the search for new counterions is of great interest. In this article, we have compiled key indexes that can be obtained from databases to guide the search for suitable counterions, together with different drug delivery system strategies to choose the most appropriate formulation according to the non-parenteral route of administration selected. Intellectual property advancements in the field are also presented and analyzed.
Assuntos
Portadores de Fármacos/administração & dosagem , Portadores de Fármacos/metabolismo , Composição de Medicamentos/métodos , Sistemas de Liberação de Medicamentos/métodos , Líquidos Iônicos/administração & dosagem , Líquidos Iônicos/metabolismo , Animais , Vias de Administração de Medicamentos , Portadores de Fármacos/síntese química , Humanos , Interações Hidrofóbicas e Hidrofílicas/efeitos dos fármacos , Líquidos Iônicos/síntese química , Permeabilidade/efeitos dos fármacosRESUMO
11-Beta hydroxysteroid dehydrogenase type 1 (11ß-HSD1) regulates cortisol levels mainly in adipose, hepatic and brain tissues. There is a relationship between the high activity of this enzyme and the development of obesity and metabolic disorders. The inhibition of 11ß-HSD1 has been shown to attenuate the development of type 2 diabetes mellitus, insulin resistance, metabolic syndrome and other diseases mediated by excessive cortisol production. In this work, fifteen benzothiazole derivatives substituted with electron-withdrawing and electron-donating groups were designed to explore their affinity for 11ß-HSD1 using in silico methods. The results show that (E)-5-((benzo[d]thiazol-2-ylimino)(methylthio)methylamino)-2-hydroxybenzoic acid (C1) has good physicochemical properties and favorable interactions with 11ß-HSD1 through hydrogen bonding and hydrophobic interactions in the catalytic site formed by Y183, S170 and Y177. Furthermore, C1 was synthesized and evaluated in vitro and ex vivo using clobenzorex (CLX) as a reference drug in obese Zucker rats. The in vitro results showed that C1 was a better inhibitor of human 11ß-HSD1 than CLX. The ex vivo assay results demonstrated that C1 was capable of reducing 11ß-HSD1 overexpression in mesenteric adipose tissue. Therefore, C1 was able to decrease the activity and expression of 11ß-HSD1 better than CLX.
Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , Benzotiazóis/química , Benzotiazóis/síntese química , Inibidores Enzimáticos/química , Inibidores Enzimáticos/síntese química , Anfetaminas/farmacologia , Animais , Benzotiazóis/farmacologia , Domínio Catalítico/efeitos dos fármacos , Diabetes Mellitus Tipo 2/tratamento farmacológico , Diabetes Mellitus Tipo 2/metabolismo , Inibidores Enzimáticos/farmacologia , Humanos , Ligação de Hidrogênio/efeitos dos fármacos , Interações Hidrofóbicas e Hidrofílicas/efeitos dos fármacos , Masculino , Simulação de Acoplamento Molecular , Obesidade/tratamento farmacológico , Obesidade/metabolismo , Ratos , Ratos ZuckerRESUMO
This study evaluated the influence of the extract of Eugenia uniflora in adhesion to human buccal epithelial cells (HBEC) biofilm formation and cell surface hydrophobicity (CSH) of Candida spp. isolated from the oral cavity of kidney transplant patients. To evaluate virulence attributes in vitro, nine yeasts were grown in the presence and absence of 1000 µg/mL of the extract. Adhesion was quantified using the number of Candida cells adhered to 150 HBEC determined by optical microscope. Biofilm formation was evaluated using two methodologies: XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) and crystal violet assay, and further analyzed by electronic scan microscopy. CSH was quantified with the microbial adhesion to hydrocarbons test. We could detect that the extract of E. uniflora was able to reduce adhesion to HBEC and CSH for both Candida albicans and non-Candida albicansCandida species. We also observed a statistically significant reduced ability to form biofilms in biofilm-producing strains using both methods of quantification. However, two highly biofilm-producing strains of Candida tropicalis had a very large reduction in biofilm formation. This study reinforces the idea that besides growth inhibition, E. uniflora may interfere with the expression of some virulence factors of Candida spp. and may be possibly applied in the future as a novel antifungal agent.
Assuntos
Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Extratos Celulares/química , Eugenia/química , Antifúngicos/química , Biofilmes/efeitos dos fármacos , Candida albicans/patogenicidade , Adesão Celular/efeitos dos fármacos , Extratos Celulares/farmacologia , Células Epiteliais/química , Células Epiteliais/efeitos dos fármacos , Humanos , Interações Hidrofóbicas e Hidrofílicas/efeitos dos fármacos , Transplante de Rim/efeitos adversos , Boca/efeitos dos fármacos , Mucosa Bucal/química , Propriedades de Superfície/efeitos dos fármacos , Fatores de Virulência/químicaRESUMO
Resveratrol can also inhibit the activation of proinflammatory mediators and cytokines at the early gene expression stage. It is well known that lectins are sugar-binding proteins that act as both pro- and anti-inflammatory molecules. Thus, the objective of this work was to verify the binding of a polyphenol compound with a lectin of Canavalia maritima (ConM) based on their ability to inhibit pro-inflammatory processes. To accomplish this, ConM was purified and crystallized, and resveratrol was soaked at 5mM for 2h of incubation. The crystal belongs to the monoclinic space group C2, the final refinement resulted in an Rfactor of 16.0% and an Rfree of 25.5%. Resveratrol binds in the rigid ß-sheet through H-bonds and hydrophobic interaction with amino acids that compose the fifth and sixth ß-strands of the rigid ß-sheet of ConM. The ConM and resveratrol inhibited DPPH oxidation, showing synergic activity with the most effective ratio of 2:3 and carbohydrate binding site is not directly related to antioxidant activity. It is the interaction between ConM and resveratrol that indicates the synergism of these two molecules in acting as free radicals scavengers and in reducing the inflammatory process through the inhibition of many pro-inflammatory events.
Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Lectinas de Plantas/química , Lectinas de Plantas/metabolismo , Polifenóis/química , Polifenóis/metabolismo , Estilbenos/química , Estilbenos/metabolismo , Sítios de Ligação , Compostos de Bifenilo/química , Canavalia , Cristalografia por Raios X , Sequestradores de Radicais Livres/farmacologia , Glicosilação/efeitos dos fármacos , Ligação de Hidrogênio/efeitos dos fármacos , Interações Hidrofóbicas e Hidrofílicas/efeitos dos fármacos , Simulação de Acoplamento Molecular , Picratos/química , Ligação Proteica/efeitos dos fármacos , Estrutura Secundária de Proteína , Quercetina/farmacologia , ResveratrolRESUMO
Staphylococcus aureus is an opportunistic pathogen often multidrug-resistant that not only causes a variety of human diseases, but also is able to survive on biotic and abiotic surfaces through biofilm communities. The best way to inhibit biofilm establishment is to prevent cell adhesion. In the present study, subinhibitory concentrations of the bacteriocins bovicin HC5 and nisin were tested for their capability to interfere with the adhesion of S. aureus to polystyrene. Subinhibitory dosages of the bacteriocins reduced cell adhesion and this occurred probably due to changes in the hydrophobicity of the bacterial cell and polystyrene surfaces. After treatment with bovicin HC5 and nisin, the surfaces became more hydrophilic and the free energy of adhesion (∆G(adhesion)) between bacteria and the polystyrene surface was unfavorable. The transcriptional level of selected genes was assessed by RT-qPCR approach, revealing that the bacteriocins affected the expression of some important biofilm associated genes (icaD, fnbA, and clfB) and rnaIII, which is involved in the quorum sensing mechanism. The conditioning of food-contact surfaces with bacteriocins can be an innovative and powerful strategy to prevent biofilms in the food industry. The results are relevant for food safety as they indicate that bovicin HC5 and nisin can inhibit bacterial adhesion and consequent biofilm establishment, since cell adhesion precedes biofilm formation.
Assuntos
Aderência Bacteriana/efeitos dos fármacos , Bacteriocinas/farmacologia , Interações Hidrofóbicas e Hidrofílicas/efeitos dos fármacos , Nisina/farmacologia , Poliestirenos , Staphylococcus aureus/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Genes Bacterianos , Humanos , Percepção de Quorum/genética , Staphylococcus aureus/metabolismoRESUMO
Smallpox was one of the most devastating diseases in the human history and still represents a serious menace today due to its potential use by bioterrorists. Considering this threat and the non-existence of effective chemotherapy, we propose the enzyme thymidylate kinase from Variola virus (VarTMPK) as a potential target to the drug design against smallpox. We first built a homology model for VarTMPK and performed molecular docking studies on it in order to investigate the interactions with inhibitors of Vaccinia virus TMPK (VacTMPK). Subsequently, molecular dynamics (MD) simulations of these compounds inside VarTMPK and human TMPK (HssTMPK) were carried out in order to select the most promising and selective compounds as leads for the design of potential VarTMPK inhibitors. Results of the docking and MD simulations corroborated to each other, suggesting selectivity towards VarTMPK and, also, a good correlation with the experimental data.
Assuntos
Modelos Moleculares , Núcleosídeo-Fosfato Quinase/química , Varíola/prevenção & controle , Vírus da Varíola/enzimologia , Sequência de Aminoácidos , Aminoácidos/metabolismo , Sítios de Ligação , Bromodesoxiuridina/metabolismo , Humanos , Ligação de Hidrogênio/efeitos dos fármacos , Interações Hidrofóbicas e Hidrofílicas/efeitos dos fármacos , Simulação de Acoplamento Molecular , Dados de Sequência Molecular , Núcleosídeo-Fosfato Quinase/antagonistas & inibidores , Inibidores de Proteínas Quinases/química , Inibidores de Proteínas Quinases/farmacologia , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Termodinâmica , Vírus da Varíola/efeitos dos fármacosRESUMO
OBJECTIVE: This study evaluated the hydrophobicity of dentin surfaces that were modified through chemical silanization with octadecyltrichlorosilane (OTS). MATERIAL AND METHODS: An in vitro experimental study was performed using 40 human permanent incisors that were divided into the following two groups: non-silanized and silanized. The specimens were pretreated and chemically modified with OTS. After the chemical modification, the dentin hydrophobicity was examined using a water contact angle measurement (WCA). The effectiveness of the modification of hydrophobicity was verified by the fluid permeability test (FPT). RESULTS AND CONCLUSIONS: Statistically significant differences were found in the values of WCA and FPT between the two groups. After silanization, the hydrophobic intraradicular dentin surface exhibited in vitro properties that limit fluid penetration into the sealed root canal. This chemical treatment is a new approach for improving the sealing of the root canal system.
Assuntos
Cavidade Pulpar/efeitos dos fármacos , Dentina/efeitos dos fármacos , Interações Hidrofóbicas e Hidrofílicas/efeitos dos fármacos , Tratamento do Canal Radicular/métodos , Silanos/química , Infiltração Dentária , Cavidade Pulpar/química , Dentina/química , Permeabilidade da Dentina/efeitos dos fármacos , Humanos , Microscopia Eletrônica de Varredura , Reprodutibilidade dos Testes , Camada de Esfregaço , Propriedades de Superfície/efeitos dos fármacos , Fatores de Tempo , Raiz Dentária/química , Raiz Dentária/efeitos dos fármacosRESUMO
Objective: This study evaluated the hydrophobicity of dentin surfaces that were modified through chemical silanization with octadecyltrichlorosilane (OTS). Material and Methods: An in vitro experimental study was performed using 40 human permanent incisors that were divided into the following two groups: non-silanized and silanized. The specimens were pretreated and chemically modified with OTS. After the chemical modification, the dentin hydrophobicity was examined using a water contact angle measurement (WCA). The effectiveness of the modification of hydrophobicity was verified by the fluid permeability test (FPT). Results and Conclusions: Statistically significant differences were found in the values of WCA and FPT between the two groups. After silanization, the hydrophobic intraradicular dentin surface exhibited in vitro properties that limit fluid penetration into the sealed root canal. This chemical treatment is a new approach for improving the sealing of the root canal system.
Assuntos
Humanos , Cavidade Pulpar/efeitos dos fármacos , Dentina/efeitos dos fármacos , Interações Hidrofóbicas e Hidrofílicas/efeitos dos fármacos , Tratamento do Canal Radicular/métodos , Silanos/química , Infiltração Dentária , Cavidade Pulpar/química , Permeabilidade da Dentina/efeitos dos fármacos , Dentina/química , Microscopia Eletrônica de Varredura , Reprodutibilidade dos Testes , Camada de Esfregaço , Propriedades de Superfície/efeitos dos fármacos , Fatores de Tempo , Raiz Dentária/química , Raiz Dentária/efeitos dos fármacosRESUMO
BACKGROUND: The pathogenesis related protein PR10 (TcPR-10), obtained from the Theobroma cacao-Moniliophthora perniciosa interaction library, presents antifungal activity against M. perniciosa and acts in vitro as a ribonuclease. However, despite its biotechnological potential, the TcPR-10 has the P-loop motif similar to those of some allergenic proteins such as Bet v 1 (Betula verrucosa) and Pru av 1 (Prunus avium). The insertion of mutations in this motif can produce proteins with reduced allergenic power. The objective of the present work was to evaluate the allergenic potential of the wild type and mutant recombinant TcPR-10 using bioinformatics tools and immunological assays. METHODOLOGY/PRINCIPAL FINDINGS: Mutant substitutions (T10P, I30V, H45S) were inserted in the TcPR-10 gene by site-directed mutagenesis, cloned into pET28a and expressed in Escherichia coli BL21(DE3) cells. Changes in molecular surface caused by the mutant substitutions was evaluated by comparative protein modeling using the three-dimensional structure of the major cherry allergen, Pru av 1 as a template. The immunological assays were carried out in 8-12 week old female BALB/c mice. The mice were sensitized with the proteins (wild type and mutants) via subcutaneous and challenged intranasal for induction of allergic airway inflammation. CONCLUSIONS/SIGNIFICANCE: We showed that the wild TcPR-10 protein has allergenic potential, whereas the insertion of mutations produced proteins with reduced capacity of IgE production and cellular infiltration in the lungs. On the other hand, in vitro assays show that the TcPR-10 mutants still present antifungal and ribonuclease activity against M. perniciosa RNA. In conclusion, the mutant proteins present less allergenic potential than the wild TcPR-10, without the loss of interesting biotechnological properties.
Assuntos
Alérgenos/imunologia , Antígenos de Plantas/imunologia , Cacau , Proteínas de Plantas/imunologia , Algoritmos , Alérgenos/química , Sequência de Aminoácidos , Animais , Antifúngicos/farmacologia , Antígenos de Plantas/química , Basidiomycota/citologia , Basidiomycota/efeitos dos fármacos , Lavagem Broncoalveolar , Cacau/química , Cacau/imunologia , Contagem de Células , Biologia Computacional , Bases de Dados de Proteínas , Feminino , Interações Hidrofóbicas e Hidrofílicas/efeitos dos fármacos , Imunoglobulina E/imunologia , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Viabilidade Microbiana/efeitos dos fármacos , Modelos Moleculares , Dados de Sequência Molecular , Proteínas Mutantes/química , Proteínas Mutantes/imunologia , Proteínas de Plantas/química , Proteínas de Plantas/farmacologia , Ribonucleases/metabolismo , Alinhamento de Sequência , Homologia Estrutural de ProteínaRESUMO
Benzaldehyde semicarbazone (BS) inhibited zymosan writhing response, carrageenan paw edema and both phases of formaldehyde nociceptive response. 2-hydroxybenzaldehyde semicarbazone (2-OHBS) and semicarbazide inhibited carrageenan paw edema and the second phase of formaldehyde nociceptive response. 2-OHBS inhibited zymosan writhing response. 3- and 4-OHBS did not show such activities. 2-OHBS showed the lowest LUMO energy, the highest contribution of the iminic carbon to LUMO energy, the highest positive charge on the iminic carbon, the highest negative charge on the iminic nitrogen and the highest susceptibility to hydrolysis. Hence semicarbazide may play important roles in 2-OHBS's activities. Inhibition of the first phase of formaldehyde response by BS could be attributed to its higher hydrophobicity and lower susceptibility to hydrolysis in comparison to 2-OHBS.
Assuntos
Analgésicos/química , Analgésicos/farmacologia , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Edema/tratamento farmacológico , Interações Hidrofóbicas e Hidrofílicas/efeitos dos fármacos , Dor/tratamento farmacológico , Semicarbazonas/síntese química , Semicarbazonas/farmacologia , Animais , Carragenina/toxicidade , Edema/induzido quimicamente , Formaldeído/toxicidade , Hidrólise/efeitos dos fármacos , Masculino , Camundongos , Estrutura Molecular , Atividade Motora/efeitos dos fármacos , Dor/induzido quimicamente , Medição da Dor , Ratos , Ratos Wistar , Semicarbazonas/químicaRESUMO
A gene for processing α-glucosidase I from a filamentous fungus, Aspergillus brasiliensis (formerly called Aspergillus niger) ATCC 9642 was cloned and fused to a glutathione S-transferase tag. The active construct with the highest production level was a truncation mutant deleting the first 16 residues of the hydrophobic N-terminal domain. This fusion enzyme hydrolyzed pyridylaminated (PA-) oligosaccharides Glc(3)Man(9)GlcNAc(2)-PA and Glc(3)Man(4)-PA and the products were identified as Glc(2)Man(9)GlcNAc(2)-PA and Glc(2)Man(4)-PA, respectively. Saturation curves were obtained for both Glc(3)Man(9)GlcNAc(2)-PA and Glc(3)Man(4)-PA, and the K (m) values for both substrates were estimated in the micromolar range. When 1 µM Glc(3)Man(4)-PA was used as a substrate, the inhibitors kojibiose and 1-deoxynojirimycin had similar effects on the enzyme; at 20 µM concentration, both inhibitors reduced activity by 50%.
Assuntos
Aspergillus/enzimologia , alfa-Glucosidases/metabolismo , Acetilglucosamina/metabolismo , Sequência de Aminoácidos , Aspergillus/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Ensaios Enzimáticos , Inibidores Enzimáticos/farmacologia , Vetores Genéticos/genética , Inibidores de Glicosídeo Hidrolases , Hidrólise/efeitos dos fármacos , Interações Hidrofóbicas e Hidrofílicas/efeitos dos fármacos , Cinética , Dados de Sequência Molecular , Oligossacarídeos/química , Oligossacarídeos/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , alfa-Glucosidases/químicaRESUMO
The antioxidant properties of the phenothiazine nucleus (PHT) associated with mitochondrial membranes and liposomes were investigated. PHT exhibited hydrophobic interaction with lipid bilayers, as shown by the quenching of excited states of 1-palmitoyl-2[10-pyran-1-yl)]-decanoyl-sn-glycero-3-phophocholine (PPDPC) incorporated in phosphatidylcholine/phosphatidylethanolamine/cardiolipin liposomes, observed even in high ionic strength; and by the spectral changes of PHT following the addition of mitochondrial membranes. Inserted into bilayers, 5 microM PHT was able to protect lipids and cytochrome c against pro-oxidant agents and exhibited spectral changes suggestive of oxidative modifications promoted by the trapping of the reactive species. In this regard, PHT exhibited the ability to scavenge DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) free radical. PHT was also able to protect rat liver mitochondria against peroxide- and iron-induced oxidative damage and consequent swelling. At the concentration range in which the antioxidant properties were observed, PHT did not cause alterations in the membrane structure and function. This study contributes to the comprehension of the correlation structure and function of phenothiazines and antioxidant properties.
Assuntos
Antioxidantes/farmacologia , Interações Hidrofóbicas e Hidrofílicas , Membranas Artificiais , Membranas Mitocondriais/efeitos dos fármacos , Fenotiazinas/farmacologia , Animais , DNA/farmacologia , Relação Dose-Resposta a Droga , Interações Hidrofóbicas e Hidrofílicas/efeitos dos fármacos , Bicamadas Lipídicas/química , Bicamadas Lipídicas/metabolismo , Lipossomos/química , Lipossomos/metabolismo , Fluidez de Membrana/efeitos dos fármacos , Fluidez de Membrana/fisiologia , Lipídeos de Membrana/metabolismo , Lipídeos de Membrana/farmacologia , Membranas Mitocondriais/química , Membranas Mitocondriais/metabolismo , Modelos Biológicos , Fenotiazinas/química , Fosfatidilcolinas/química , Fosfatidilcolinas/farmacologia , Proteínas/efeitos dos fármacos , Proteínas/metabolismo , RatosRESUMO
A major concern in the antiretroviral (ARV) treatment of HIV infections with protease inhibitors (PI) is the emergence of resistance, which results from the selection of distinct mutations within the viral protease (PR) gene. Among patients who do not respond to treatment with the PI nelfinavir (NFV), the D30N mutation is often observed. However, several reports have shown that D30N emerges with different frequencies in distinct HIV-1 genetic forms or subtypes. In the present work, we analyzed the binding of NFV and the Gag substrate CA/p2 to PR from HIV-1 subtypes B and C through molecular dynamics (MD) simulations. The wild-type and drug-resistant D30N mutants were investigated in both subtypes. The compensatory mutations N83T and N88D, observed in vitro and in vivo when subtype C acquires D30N, were also studied. D30N appears to facilitate conformational changes in subtype B PR, but not in that from subtype C, and this could be associated with disestablishment of an alpha-helical region of the PR. Furthermore, the total contact areas of NFV or the CA/p2 substrate with the mutant PR correlated with changes in the resistance patterns and replicative capacity. Finally, we observed in our MD simulations that mutant PR proteins show different patterns for hydrophobic/van der Waals contact. These findings suggest that different molecular mechanisms contribute to resistance, and we propose that a single mutation has distinct impacts on different HIV-1 subtypes.