RESUMO
As part of our ongoing studies involving the discovery of new natural prototypes with antiprotozoal activity against Trypanosoma cruzi from Brazilian plant species, the chromatographic fractionation of hexane extract from leaves of Nectandra barbellata afforded one new pseudo-disesquiterpenoid, barbellatanic acid. The structure of this compound was elucidated by NMR and HR-ESIMS data analysis. Barbellatanic acid displayed a trypanocidal effect with IC50 of 13.2 µM to trypomastigotes and no toxicity against NCTC cells (CC50 > 200 µM), resulting in an SI value higher than 15.1. The investigation of the lethal mechanism of barbellatanic acid in trypomastigotes, using both fluorescence microscopy and spectrofluorimetric analysis, revealed a time-dependent permeation of the plasma membrane. Based on these results, this compound was incorporated in cellular membrane models built with lipid Langmuir monolayers. The interaction of barbellatanic acid with the models was inferred by tensiometric, rheological, spectroscopical, and morphological techniques, which showed that this compound altered the thermodynamic, viscoelastic, structural, and morphological properties of the film. Taking together, these results could be employed when this prodrug interacts with lipidic interfaces, such as protozoa membranes or liposomes for drug delivery systems.
Assuntos
Antiprotozoários , Tripanossomicidas , Trypanosoma cruzi , Tripanossomicidas/farmacologia , Tripanossomicidas/química , Antiprotozoários/farmacologia , Membrana Celular , Folhas de PlantaRESUMO
Biseugenol (1), a neolignan with antiprotozoal activity against Trypanosoma cruzi, was partially methylated, and the compound obtained - methyl biseugenol (2) - had its activity evaluated against the extracellular (trypomastigotes) and intracellular (amastigotes) forms of T. cruzi. It was observed that both compounds 1 and 2 exhibited similar effects against trypomastigotes (IC50 of 11.7 and 16.2 µM, respectively), whereas compound 2 displayed higher activity against amastigotes (IC50 = 8.2 µM) in comparison with biseugenol (IC50 = 15.4 µM). Additionally, reduced toxicity against NCTC cells for compound 2 was observed (CC50 > 200 µM), differently from compound 1 with CC50 = 58.0 µM. Aiming to understand better the molecular mechanism of the biological action of compound 2, the prodrug was incorporated into cellular membrane models constituted of Langmuir monolayers of the lipids dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylethanolamine (DPPE), dipalmitoylphosphatidylserine (DPPS), and dipalmitoylphosphatidylglycerol (DPPG). The lipid-drug interaction was inferred through tensiometry, surface potential, infrared spectroscopy (PM-IRRAS), and Brewster angle microscopy (BAM). The prodrug expanded DPPC and DPPG monolayers and condensed DPPE ones, as well as presented characteristic behaviors regarding the chemical structure of the lipid considering expansion-compression curves, surface potential-area isotherms, and stability of previously compressed monolayers to relevant-biological surface pressures. PM-IRRAS indicated a molecular disorder for DPPC and DPPS alkyl chains in the presence of the drug. BAM revealed the presence of domains in the DPPG and DPPE monolayers, which was probably induced by the prodrug. These data suggest, in general, that the lipid composition modulates the interaction of compound 2, whose results are expected to correlate to its trypanocidal activity, which involves the plasma membrane of T. cruzi as the primary target, i.e., the first barrier that the compound should encounter to interact with the microorganism.
Assuntos
Pró-Fármacos , Metilação , Membrana Celular/química , 1,2-Dipalmitoilfosfatidilcolina/química , Propriedades de SuperfícieRESUMO
Lysicamine, an alkaloid with tumorigenic activity, was incorporated in cell membrane models made of lipid Langmuir monolayers. Dipalmitoylphosphocholine (DPPC), dioleoylphosphocholine (DOPC), and palmitoyloleoylcholine (POPC) represented non-tumorigenic cell membranes, and dipalmitoylphosphoserine (DPPS), dioleoylphosphoserine (DOPS), and palmitoyloleoylserine (POPS), tumorigenic ones. The monolayers were characterized by tensiometry, infrared spectroscopy, and Brewster Angle Microscopy (BAM). No significant shifts of the isotherms were observed for the saturated lipids (DPPC and DPPS), while for the others (DOPC, POPS, DOPS, and POPS), more significant changes were observed not only in the compression isotherms but also in the surface pressure-time curve for pre-compressed monolayers. The molecular organization, as well as the morphology of the drug-lipid monolayers, could be inferred with infrared spectroscopy and BAM. While the first revealed that the alkyl chain ordering changed upon lysicamine incorporation, the second showed how the drug could distinctly change the state of aggregation of molecular domains at the air-water interface. In conclusion, lysicamine could interact distinctly with each lipid at the air-water interface, showing the dependence not only on the lipid polar groups but also on the level of unsaturation of the alkyl chains.
Assuntos
Fosfatidilgliceróis , Água , Água/química , Propriedades de Superfície , Membrana Celular/química , 1,2-Dipalmitoilfosfatidilcolina/químicaRESUMO
Knowing how a bioactive compound interacts with cell membranes is important to understand its effect at the molecular level. In this sense, this work aimed to study the interaction of lysicamine, an alkaloid with action against lung cancer cell lines, with lipid monolayers as cell membrane models. We employed two lipid mixtures: the first composed of 35% DOPC, 30% DOPE, 20% sphingomyelin, and 15% cholesterol as healthy cell membranes models (MM1), and the second replacing DOPC with DOPS as cancer cells models (MM2). The interaction of lysicamine with the monolayers was evaluated using tensiometry, Brewster angle microscopy (BAM), and polarization-modulated infrared reflection-absorption spectroscopy (PM-IRRAS). Lysicamine had interfacial effects in both membrane models. For MM 1, it expanded the lipid monolayer and changed the interfacial rheological properties, increasing the in-plane elasticity of the films. PM-IRRAS spectra suggested a higher conformational disorder of the alkyl chains of the lipids. For MM 2, lysicamine also shifted the isotherms to higher areas, expanding the monolayers, but with no significant alteration in their interfacial rheological properties. PM-IRRAS spectra also suggested higher disorder in the orientation of the lipid alkyl chains upon lysicamine incorporation. For both models, BAM did not show alteration in interfacial aggregation upon drug incorporation. In conclusion, changes in some interfacial properties of membrane models caused by lysicamine depend on the monolayer composition, which can be associated with its bioactivity in cellular membranes.
Assuntos
Esfingomielinas , Água , Água/química , Espectrofotometria Infravermelho , Membrana Celular , Esfingomielinas/química , Propriedades de SuperfícieRESUMO
Dehydrodieugenol, a neolignan isolated from the Brazilian plant Nectandra leucantha (Lauraceae) with reported antiprotozoal and anticancer activity, was incorporated in Langmuir monolayers of selected lipids as cell membrane models, aiming to comprehend its action mechanism at the molecular level. The interaction of this compound with the lipids dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylethanolamine (DPPE), dipalmitoylphosphatidylserine (DPPS), and dipalmitoylphosphatidylglycerol (DPPG) was inferred through tensiometry, infrared spectroscopy, and Brewster angle microscopy. The interactions had different effects depending on the chemical nature of the lipid polar head, with expansion for DPPC monolayers, condensation for DPPE, and expansion (at low surface pressures) followed by the overlap of the isotherms (at high surface pressure values) for DPPS and DPPG. Effects caused by dehydrodieugenol in the negatively charged lipids were distinctive, which was also reflected in the hysteresis assays, surface potential-area isotherms, and rheological measurements. Infrared spectroscopy indicated that the drug interaction with the monolayer affects not only the polar groups, but also the acyl lipid chains for all lipids. These results pointed to the fact that the interaction of the drug with lipid monolayers at the air-water interface is modulated by the lipid composition, mainly considering the polar head of the lipids, as well as the hydrophobicity of the lipids and the drug. As negatively charged lipids pointed to distinctive interaction, we believe this can be related to the antiprotozoal and anticancer properties of the compound.
Assuntos
Lauraceae , Lignanas , Membrana Celular/química , Eugenol/análogos & derivados , Eugenol/análise , Lignanas/análiseRESUMO
Sakuranetin, a natural compound with activity in lipidic biointerfaces, was isolated from Baccharis retusa and studied with two models of lipid membranes: Langmuir monolayers and Molecular Simulation. For that, the mammalian lipid DPPC was chosen. Sakuranetin condensed the monolayers at high surface pressures, decreased the surface compressional modulus, reduced the molecular order of the acyl chains (diminution of all-trans/gauche conformers ratio), and increased the heterogeneity of the interface, forming aggregates. Molecular simulation data gave information on the bioactive compound's most favorable thermodynamic positions along the lipid monolayer, which was the lipid-air interface. These combined results lead to the conclusion that this lipophilic compound may interact with the lipidic layers, preferentially at the lipid-air interface, to minimize the free energy, and reaches this conformation disturbing the thermodynamic, structural, mechanical, rheological, and morphological properties of the well-packed DPPC monolayer.
Assuntos
1,2-Dipalmitoilfosfatidilcolina , Lipídeos , 1,2-Dipalmitoilfosfatidilcolina/química , Membrana Celular/química , Flavonoides , Propriedades de Superfície , Termodinâmica , Água/químicaRESUMO
The present work evaluated the antiprotozoal activity of isolinderanolide E, isolated from the Brazilian plant Nectandra oppositifolia, against promastigote forms of Leishmania (Leishmania) amazonensis. The compound exhibited an EC50 value of 20.3 µM, similar to the positive control miltefosine (IC50 of 19.4 µM), and reduced toxicity to macrophages (CC50 > 200 µM). Based on these results, Langmuir monolayers of two unsaturated lipids: 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), were employed as a model of mammalian and parasite membranes, respectively, to study the interaction of isolinderanolide E at a molecular level. The films were characterized with tensiometry (surface pressure-area isotherms and surface pressure-time curves), infrared spectroscopy, and Brewster angle microscopy (BAM). This compound changed the profile of the isotherms leading to fluid DOPC and DOPE monolayers, which were not able to attain rigid states even with compression. Infrared spectroscopy showed that the bioactive compound decreases the trans/gauche ratio conformers related to the molecular conformational disorder. BAM showed the formation of specific aggregates upon drug incorporation. In conclusion, isolinderanolide E changes the thermodynamic, mechanical, structural, and morphological characteristics of the monolayer of these unsaturated lipids, which may be essential to understand the action at the molecular level bioactives in biointerfaces.
Assuntos
Antiprotozoários , Lauraceae , Animais , Antiprotozoários/farmacologia , Membrana Celular , Lipídeos/análise , Mamíferos , Propriedades de Superfície , TermodinâmicaRESUMO
A long-tail lactone, named isolinderanolide E, was obtained from Nectandra oppositifolia and incorporated in Langmuir monolayers of dipalmitoyl-phosphoethanolamine (DPPE) as a model of microbial membranes. The compound was dissolved in chloroform and mixed with DPPE to provide mixed solutions spread on the air-water interface. After solvent evaporation, mixed monolayers were formed, and surface pressure-area isotherms, dilatational rheology, Brewster angle microscopy (BAM), and infrared spectroscopy were employed to characterize the prodrug-membrane interactions. Isolinderanolide E expanded DPPE monolayers, denoting repulsive interactions. At 30 mN/m, the monolayer presented higher viscoelastic and in-plane elasticity parameters and an increased ratio of all-trans/gauche conformers of the alkyl chains, confirming molecular order. Morphology of the monolayer was analyzed by BAM, which revealed a more homogeneous distribution of Isolinderanolide E along the DPPE monolayer than the prodrug directly spread at the interface, which tends to aggregate. A molecular model proposing the molecular orientation of the amphiphilic drug is presented and explained by the distortion of the alkyl chains as well as by viscoelastic changes. In conclusion, the prodrug changes the thermodynamic, rheological, morphological, and structural properties of the DPPE monolayer, which may be essential to understand, at the molecular level, the action of bioactives in selected membrane models.
Assuntos
Anti-Infecciosos/metabolismo , Lactonas/metabolismo , Lauraceae/metabolismo , Membranas Artificiais , Modelos Químicos , Microscopia/métodos , Éteres Fenílicos/metabolismo , Reologia , Análise Espectral/métodos , TermodinâmicaRESUMO
In this paper, we studied how different hydrophilicity degrees of the polar groups of the lipids dipalmitoylphosphatidylcholine (DPPC) and dipalmitoyl phosphatidylethanolamine (DPPE) influence the interaction of the antibiotic peptide vancomycin (VC), affecting the physicochemical properties of the monolayers, including thermodynamic, rheological, structural and morphological ones. Lipid Langmuir monolayers were prepared at air-water interfaces with VC aqueous solution as subphase and characterized with tensiometry, Brewster angle microscopy, infrared spectroscopy, dilatational, and interfacial shear rheology. The presence of PC or PE groups as polar head groups of the phospholipid monolayers modulated the interaction of VC adsorbing from the aqueous subphase since for DPPC, vancomycin condenses the monolayer, making it less stable, fluid, and more disordered. In contrast, for DPPE, vancomycin expands the monolayer, making it more stable, keeping the compressibility, and leading to the formation of interfacial aggregates, which are not observed for DPPC. We concluded thatelectrostatic interactions induced the insertion of the peptide into the polar heads of the monolayers (DPPE), while hydrophobic interactions, in addition to ion-dipole interactions, induced the adsorption of the peptide onto the polar head of the monolayers (DPPC).
Assuntos
Água , Elasticidade , Fosfolipídeos , TermodinâmicaRESUMO
A known monoterpene, named γ-terpineol, was incorporated in mixed Langmuir monolayers composed of dipalmitoyl-phosphoethanolamine (DPPE) and peptidoglycans as a model of microbial membranes. Surface pressure and surface potential isotherms, dynamical surface rheology, Brewster angle microscopy (BAM), and infrared spectroscopy were employed to characterize the compound-membrane interactions. The compound expanded the monolayers denoting repulsive interactions. At 30 mN/m, the monolayer presented lower viscoelastic and in-plane elasticity parameters and an increased all-trans/gauche conformers ratio for the alkyl chains, confirming molecular order. The morphology of the monolayer was analyzed by BAM, which revealed a heterogeneous distribution of γ-terpineol along the mixed monolayer, which tends to segregate. In conclusion, the compound changes the thermodynamic, electric, rheological, morphological, and structural properties of the peptidoglycan-DPPE monolayer, which may be essential to understand, at the molecular level, the action of bioactives in selected membrane models.