RESUMEN
The susceptibility of lysosomal membranes in tumor cells to cationic amphiphilic drugs (CADs) enables CADs to induce lysosomal membrane permeabilization (LMP) and trigger lysosome-dependent cell death (LDCD), suggesting a potential antitumor therapeutic approach. However, the existence of intrinsic lysosomal damage response mechanisms limits the display of the pharmacological activity of CADs. In this study, we report that low concentrations of QS-21, a saponin with cationic amphiphilicity extracted from Quillaja Saponaria tree, can induce LMP but has nontoxicity to tumor cells. QS-21 and MAP30, a type I ribosome-inactivating protein, synergistically induce apoptosis in tumor cells at low concentrations of both. Mechanistically, QS-21-induced LMP helps MAP30 escape from endosomes or lysosomes and subsequently enter the endoplasmic reticulum, where MAP30 downregulates the expression of autophagy-associated LC3 proteins, thereby inhibiting lysophagy. The inhibition of lysophagy results in the impaired clearance of damaged lysosomes, leading to the leakage of massive lysosomal contents such as cathepsins into the cytoplasm, ultimately triggering LDCD. In summary, our study showed that coadministration of QS-21 and MAP30 amplified the lysosomal disruption and can be a new synergistic LDCD-based antitumor therapy.
Asunto(s)
Apoptosis , Autofagia , Lisosomas , Saponinas , Lisosomas/efectos de los fármacos , Lisosomas/metabolismo , Saponinas/farmacología , Apoptosis/efectos de los fármacos , Humanos , Autofagia/efectos de los fármacos , Línea Celular Tumoral , Animales , Sinergismo Farmacológico , Proteínas Inactivadoras de Ribosomas Tipo 1/farmacología , Ratones , Quillaja/química , Antineoplásicos/farmacologíaRESUMEN
Extracts of the Chilean soapbark tree, Quillaja Saponaria (QS) are the source of potent immune-stimulatory saponin compounds. This study compared the adjuvanticity and toxicity of QS-18 and QS-21, assessing the potential to substitute QS-18 in place of QS-21 for vaccine development. QS-18, the most abundant QS saponin fraction, has been largely overlooked due to safety concerns. We found that QS-18 spontaneously inserted into liposomes, thereby neutralizing hemolytic activity, and following administration did not induce local reactogenicity in a footpad swelling test in mice. With high-dose intramuscular administration, transient weight loss was minor, and QS-18 did not induce significantly more weight loss compared to a liposome vaccine adjuvant system lacking it. Two days after administration, no elevation of inflammatory cytokines was detected in murine serum. In a formulation including cobalt-porphyrin-phospholipid (CoPoP) for short peptide sequestration, QS-18 did not impact the formation of peptide nanoparticles. With immunization, QS-18 peptide particles induced higher levels of cancer neoepitope-specific and tumor-associated antigen-specific CD8+ T cells compared to QS-21 particles, without indication of greater toxicity based on mouse body weight. T cell receptor sequencing of antigen-specific CD8+ T cells showed that QS-18 induced significantly more T cell transcripts. In two murine cancer models, vaccination with QS-18 peptide particles induced a similar therapeutic effect as QS-21 particles, without indication of increased toxicity. Antigen-specific CD8+ T cells in the tumor microenvironment were found to express the exhaustion marker PD-1, pointing to the rationale for exploring combination therapy. Taken together, these data demonstrate that QS-18, when formulated in liposomes, can be a safe and effective adjuvant to induce tumor-inhibiting cellular responses in murine models with potential to facilitate or diminish costs of production for vaccine adjuvant systems. Further studies are warranted to assess liposomal QS-18 immunogic, reactogenic and toxicological profiles in mice and other animal species.
Asunto(s)
Adyuvantes Inmunológicos , Vacunas contra el Cáncer , Liposomas , Quillaja , Animales , Vacunas contra el Cáncer/administración & dosificación , Vacunas contra el Cáncer/inmunología , Quillaja/química , Adyuvantes Inmunológicos/administración & dosificación , Femenino , Ratones Endogámicos C57BL , Vacunas de Subunidad/administración & dosificación , Vacunas de Subunidad/inmunología , Ratones , Saponinas de Quillaja , Citocinas , Saponinas/administración & dosificación , Saponinas/farmacología , Línea Celular Tumoral , Vacunas de Subunidades ProteicasRESUMEN
BACKGROUND: Malaria is preventable yet causes >600 000 deaths annually. RTS,S, the first marketed malaria vaccine, has modest efficacy, but improvements are needed for eradication. METHODS: We conducted an open-label, dose escalation phase 1 study of a full-length recombinant circumsporozoite protein vaccine (rCSP) administered with adjuvant glucopyranosyl lipid A-liposome Quillaja saponaria 21 formulation (GLA-LSQ) on days 1, 29, and 85 or 1 and 490 to healthy, malaria-naive adults. The primary end points were safety and reactogenicity. The secondary end points were antibody responses and Plasmodium falciparum parasitemia after homologous controlled human malaria infection. RESULTS: Participants were enrolled into 4 groups receiving rCSP/GLA-LSQ: 10â µg × 3 (n = 20), 30â µg × 3 (n = 10), 60â µg × 3 (n = 10), or 60â µg × 2 (n = 9); 10 participants received 30â µg rCSP alone × 3, and there were 6 infectivity controls. Participants experienced no serious adverse events. Rates of solicited and unsolicited adverse events were similar among groups. All 26 participants who underwent controlled human malaria infection 28 days after final vaccinations developed malaria. Increasing vaccine doses induced higher immunoglobulin G titers but did not achieve previously established RTS,S benchmarks. CONCLUSIONS: rCSP/GLA-LSQ had favorable safety results. However, tested regimens did not induce protective immunity. Further investigation could assess whether adjuvant or schedule adjustments improve efficacy. CLINICAL TRIALS REGISTRATION: NCT03589794.
Asunto(s)
Adyuvantes Inmunológicos , Anticuerpos Antiprotozoarios , Lípido A , Liposomas , Vacunas contra la Malaria , Malaria Falciparum , Plasmodium falciparum , Proteínas Protozoarias , Humanos , Vacunas contra la Malaria/inmunología , Vacunas contra la Malaria/administración & dosificación , Vacunas contra la Malaria/efectos adversos , Malaria Falciparum/prevención & control , Malaria Falciparum/inmunología , Adulto , Plasmodium falciparum/inmunología , Proteínas Protozoarias/inmunología , Femenino , Masculino , Adyuvantes Inmunológicos/administración & dosificación , Adulto Joven , Lípido A/análogos & derivados , Lípido A/administración & dosificación , Lípido A/inmunología , Anticuerpos Antiprotozoarios/sangre , Anticuerpos Antiprotozoarios/inmunología , Quillaja/química , Adolescente , Vacunas Sintéticas/inmunología , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/efectos adversos , Persona de Mediana Edad , GlucósidosRESUMEN
Modification of the 3-glucuronic acid (GlcA) residue from the Quillaja saponin (QS) adjuvants by N-acylation, yields derivatives with linear alkylamides that show structural and functional changes. Structural, since the relatively unreactive added hydrophobic alkyl chains may modify these glycosides' conformation and micellar structure. Functional, because altering the availability of proposed pharmacophores, like fucose (Fucp) and aldehyde groups, to interact with their cellular receptors, may change these glycosides' adjuvanticity. While deacylated QS (DS-QS) adjuvants bias the response toward a sole anti-inflammatory Th2 immunity against an antigen, their N-alkylated derivatives carrying octyl to dodecylamide residues, modify that response to a pro-inflammatory Th1 immunity. As shown by their IgG2a/IgG1 titer ratios, which are higher than those for Th2 immunity. A result of the fact that in mice, the IgG2a levels are dependent on the direct influence of secreted interferon-γ (IFN-γ), a crucial Th1 cytokine. But addition of the longer and more lipophilic tetradecylamide group, yields derivatives that like DS-QS induce Th2 immunity, as shown by their low IgG2a/IgG1 ratio. Results that imply that changes in these analogs' conformation and micellar structure, would affect the immunomodulatory properties or adjuvanticity of N-acylated DS-QS. Physical changes that may alter the availability of groups like Fucp, to bind to its presumed dendritic cells' lectin receptor DC-SIGN; an essential step in the stimulation of Th2 immunity. Structural properties that in an aqueous environment, would depend on these glycosides' balance of their hydrophilic and lipophilic moieties (HLB), and the interactions of the newly introduced alkyl chain with the native QS' lipophilic triterpene aglycone and hydrophilic oligosaccharide chains. A situation that would explain these new derivatives' qualitative and quantitative changes in adjuvanticity.
Asunto(s)
Saponinas , Acilación , Adyuvantes Inmunológicos/química , Animales , Inmunoglobulina G , Ratones , Quillaja/química , Saponinas de QuillajaRESUMEN
Triterpene saponins of the genus Quillaja (Quillajaceae) are known for their immunoadjuvant, hypocholesterolemic, and anti-inflammatory activity. Plant cell cultures are useful for the study of saponin metabolism and industrial production of these bioactive compounds. While structurally related phytosterols are primary metabolites essential to growth and development, saponins are responsive to pathogen and abiotic stress, fulfilling roles in plant specialized metabolism. For cell culture production of saponins, phytosterols may be considered a competing pathway which relies on a common pool of cytosolic isoprenoid precursors.Understanding the metabolic allocation of resources between these two related pathways is key to maximizing saponin production in in vitro production systems. Sterols and saponins naturally occur in multiple conjugated forms, which complicate separation and quantification. The acid hydrolysis of conjugated sterols and saponins to their free forms is a useful technique to simplify their analysis by gas chromatography. Here we provide the workflow for the quantification of free sterols and sapogenins in cell cultures of Quillaja brasiliensis .
Asunto(s)
Fitosteroles , Sapogeninas , Saponinas , Triterpenos , Técnicas de Cultivo de Célula , Cromatografía de Gases y Espectrometría de Masas , Quillaja/química , Saponinas/química , EsterolesRESUMEN
An immunoadjuvant preparation (named Fraction B) was obtained from the aqueous extract of Quillaja brasiliensis leaves, and further fractionated by consecutive separations with silica flash MPLC and reverse phase HPLC. Two compounds were isolated, and their structures elucidated using a combination of NMR spectroscopy and mass spectrometry. One of these compounds is a previously undescribed triterpene saponin (Qb1), which is an isomer of QS-21, the unique adjuvant saponin employed in human vaccines. The other compound is a triterpene saponin previously isolated from Quillaja saponaria bark, known as S13. The structure of Qb1 consists of a quillaic acid residue substituted with a ß-d-Galp-(1â2)-[ß-d-Xylp-(1â3)]-ß-d-GlcpA trisaccharide at C3, and a ß-d-Xylp-(1â4)-α-l-Rhap-(1â2)-[α-l-Arap-(1â3)]-ß-d-Fucp moiety at C28. The oligosaccharide at C28 was further substituted at O4 of the fucosyl residue with an acyl group capped with a ß-d-Xylp residue.
Asunto(s)
Saponinas , Triterpenos , Adyuvantes Inmunológicos/química , Humanos , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Quillaja/química , Saponinas/química , Triterpenos/químicaRESUMEN
Nanoadjuvants that combine immunostimulatory properties and delivery systems reportedly bestow major improvements on the efficacy of recombinant, protein-based vaccines. Among these, self-assembled micellar formulations named ISCOMs (immune stimulating complexes) show a great ability to trigger powerful immunological responses against infectious pathogens. Here, a nanoadjuvant preparation, based on saponins from Quillaja brasiliensis, was evaluated together with an experimental Zika virus (ZIKV) vaccine (IQB80-zEDIII) and compared to an equivalent vaccine with alum as the standard adjuvant. The preparations were administered to mice in two doses (on days zero and 14) and immune responses were evaluated on day 28 post-priming. Serum levels of anti-Zika virus IgG, IgG1, IgG2b, IgG2c, IgG3 were significantly increased by the nanoadjuvant vaccine, compared to the mice that received the alum-adjuvanted vaccine or the unadjuvanted vaccine. In addition, a robust production of neutralizing antibodies and in vitro splenocyte proliferative responses were observed in mice immunized with IQB80-zEDIII nanoformulated vaccine. Therefore, the IQB80-zEDIII recombinant preparation seems to be a suitable candidate vaccine for ZIKV. Overall, this study identified saponin-based delivery systems as an adequate adjuvant for recombinant ZIKV vaccines and has important implications for recombinant protein-based vaccine formulations against other flaviviruses and possibly enveloped viruses.
Asunto(s)
Adyuvantes Inmunológicos , ISCOMs/inmunología , Quillaja/química , Saponinas/inmunología , Proteínas del Envoltorio Viral/inmunología , Vacunas Virales/inmunología , Virus Zika/inmunología , Adyuvantes Inmunológicos/administración & dosificación , Animales , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/inmunología , ISCOMs/administración & dosificación , Inmunogenicidad Vacunal , Linfocitos/inmunología , Ratones , Dominios Proteicos , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Saponinas/química , Proteínas del Envoltorio Viral/química , Proteínas del Envoltorio Viral/genética , Vacunas Virales/administración & dosificaciónRESUMEN
QS-21 is a triterpene glycoside saponin found in the bark of the Chilean soap bark tree Quillaja saponaria. It is a highly potent vaccine adjuvant that is included in two approved vaccines and has shown promise in numerous other vaccine candidates in the research and clinical pipelines. One major hurdle to the widespread use of this adjuvant is the difficulty of obtaining it in high yield and purity. Previously reported purification approaches either showed suboptimal purity and/or yield, lacked efficiency, or had strict requirement on the composition of the starting material. Here, we report the development of a new two-step orthogonal chromatographic process, consisting of a polar reversed-phase (RP) chromatography step followed by a hydrophilic interaction chromatography (HILIC) step, for purifying QS-21 from a commercially available Quillaja saponaria bark extract with high yield and > 97% purity. This process makes available a simple and efficient method for obtaining highly pure QS-21 from saponin-enriched bark extract.
Asunto(s)
Cromatografía/métodos , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/normas , Saponinas/aislamiento & purificación , Saponinas/normas , Chile , Extractos Vegetales/química , Quillaja/químicaRESUMEN
A series of studies was carried out to determine the anticoccidial effects of a product derived from plant material sourced from Quillaja saponaria and Yucca schidigera. These plants are known to contain high concentrations of triterpenoid and steroidal saponins, substances that are known to display an array of biological effects. Battery tests involving individual Eimeria acervulina, Eimeria maxima, and Eimeria tenella infections and graded levels of a quillaja/yucca combination (QY) (0, 200, 250, and 300 ppm) were conducted. Body weight gain, coccidial lesion scores, and total oocysts per gram of feces (OPG) were used to evaluate anticoccidial effects. In addition, three floor pen trials evaluated the effects of 250 ppm QY in the control coccidial infections. The first pen trial measured the effects of 250 ppm QY, both alone and in combination with 66 ppm salinomycin (Sal), in a 2 3 2 factorial treatment arrangement. Two additional 42-day pen studies assessed the effects 250 ppm QY in birds vaccinated for coccidiosis. Data from the three battery trials indicated that at doses of 250 ppm QY or more, weight gain was improved, E. acervulina and E. tenella lesion scores were reduced, and OPG was lowered. In general, OPG was reduced by about 50% across all species by 250 and 300 ppm QY. Results of the pen study indicated that 250 ppm QY and Sal, when fed individually, reduced OPG and lesion scores and improved final performance. However, when QY and Sal were administered concurrently, further significant reductions in OPG occurred. The final performance of broilers vaccinated for coccidiosis was also improved at 250 ppm QY, as was OPG at both 21 and 28 days. Thus, at QY doses of 250 ppm or more, anticoccidial activity was evident but lacked the potency exhibited by many standard anticoccidials. When combined with either Sal or a live coccidiosis vaccine, QY improved the anticoccidial effects and performance of these anticoccidial methods.
Asunto(s)
Pollos , Coccidiosis/veterinaria , Coccidiostáticos/metabolismo , Eimeria/efectos de los fármacos , Enfermedades de las Aves de Corral/prevención & control , Quillaja/química , Saponinas/metabolismo , Yucca/química , Alimentación Animal/análisis , Animales , Coccidiosis/prevención & control , Coccidiostáticos/administración & dosificación , Dieta/veterinaria , Suplementos Dietéticos/análisis , Relación Dosis-Respuesta a Droga , Piranos/administración & dosificación , Distribución Aleatoria , Saponinas/administración & dosificaciónRESUMEN
Saponins from Quillaja saponaria have been commonly used as immunomodulatory adjuvants in foot-and-mouth disease vaccines (FMDVs). However, due to the lack of consensus over the possible exacerbation of local inflammatory responses in cattle and its economic impacts, their use has been discouraged by Brazilian authorities. A qualitative method intended to determine the presence of saponins from Q. saponaria bark extracts in FMDVs was developed and validated. Instrumental analysis was performed using an liquid chromatography (LC) coupled to a quadrupole-time-of-flight-mass spectrometry (TOF-MS) system. The method was validated according to the International Conference on Harmonization Harmonized Tripartite Guideline Q2 (R1) and Brazilian Ministry of Agriculture, Livestock and Food Supply Analytical Quality Assurance Guidelines. Validation parameters were determined and considered suitable to the established criteria. The validated method has been applied in routine analysis in the National Agricultural Laboratory at Rio Grande do Sul (LANAGRO-RS). All results obtained were in agreement with the vaccine's composition described by the manufacturer. The method is easy and adequate for analysis in routine laboratories. To the best of the authors' knowledge, this is the first report of a method which intends to investigate the presence of saponins from Q. saponaria bark extracts in veterinary vaccines.
Asunto(s)
Adyuvantes Inmunológicos/química , Cromatografía Liquida/métodos , Quillaja/química , Saponinas/análisis , Vacunas Virales/química , Animales , Fiebre Aftosa/prevención & control , Espectrometría de Masas/métodos , Corteza de la Planta/química , Extractos Vegetales/química , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
Replacing synthetic surfactants by natural alternatives when formulating nanoemulsions has gained attention as a sustainable approach. In this context, nanoemulsions based on sweet almond oil and stabilized by saponin from Quillaja bark with glycerol as cosurfactant were prepared by the high-pressure homogenization method. The effects of oil/water (O/W) ratio, total surfactant amount, and saponin/glycerol ratio on their stability were analyzed. The formation and stabilization of the oil-in-water nanoemulsions were analyzed through the evaluation of stability over time, pH, zeta potential, and particle size distribution analysis. Moreover, a design of experiments was performed to assess the most suitable composition based on particle size and stability parameters. The prepared nanoemulsions are, in general, highly stable over time, showing zeta potential values lower than -40 mV, a slight acid behavior due to the character of the components, and particle size (in volume) in the range of 1.1 to 4.3 µm. Response surface methodology revealed that formulations using an O/W ratio of 10/90 and 1.5 wt% surfactant resulted in lower particle sizes and zeta potential, presenting higher stability. The use of glycerol did not positively affect the formulations, which reinforces the suitability of preparing highly stable nanoemulsions based on natural surfactants such as saponins.
Asunto(s)
Emulsiones/química , Nanoestructuras/química , Corteza de la Planta/química , Quillaja/química , Saponinas/química , Tensoactivos/química , Dispersión Dinámica de Luz , Glicerol/química , Concentración de Iones de Hidrógeno , Tamaño de la Partícula , Aceites de Plantas/química , Agua/químicaRESUMEN
Knowledge of binary emulsifiers' influence on the formation and stability of emulsion-based products is still limited. The aim of this study was to investigate the emulsifying properties of Quillaja saponin-egg lecithin mixtures at different concentration ratios (r = 5:0, 4:1, 3:2, 2:3, 1:4, and 0:5) with total emulsifier concentration set to 0.5% or 1.0% (w/w). For this, oil-in-water emulsions (10% oil, pH 7) were prepared via high-pressure homogenization. Furthermore, emulsion stability against different environmental stresses was tested. All the binary emulsifier mixtures formed submicron sized emulsions upon homogenization. The most stable emulsions among the mixed emulsifiers were obtained at low Quillaja saponin concentration at r = 1:4 that showed similar physical stability over time to emulsions stabilized by Quillaja saponins and egg lecithin alone. The data suggested that the mixtures of Quillaja saponins and egg lecithins built mixed interfacial layers that were prone to changes over time. Emulsions stabilized by the binary mixtures were in general less stable against changes in pH and ionic strength than the emulsions stabilized by the individual emulsifiers. An exception were the emulsions at r = 1:4 that showed improved stability at pH 2 over the phase separated Quillaja saponin-stabilized emulsions at the same pH. Moreover, all the emulsions were heat stable up to 90 °C. On the other hand, none of the emulsions were stable upon freeze-thawing. These results increase our understanding of technofunctionality of binary emulsifier systems. PRACTICAL APPLICATION: Food-grade and natural emulsifier mixtures composed of Quillaja saponins and egg lecithin may be used in selected emulsion-based food or personal care product applications to replace synthetic surfactants due to issues with consumer acceptance and regulatory restrictions.
Asunto(s)
Lecitinas/química , Extractos Vegetales/química , Quillaja/química , Emulsionantes/química , Emulsiones/química , Concentración Osmolar , Tamaño de la Partícula , Saponinas de Quillaja/química , Tensoactivos/químicaRESUMEN
Saponins are known for their bioactive and surfactant properties, showing applicability to the food, cosmetic and pharmaceutical industries. This work evaluated the saponins effects on Kluyveromyces lactis ß-galactosidase activity and correlated these changes to the protein structure. Enzyme kinetic was evaluated by catalytic assay, protein structure was studied by circular dichroism and fluorescence, and isothermal titration calorimetry was used to evaluate the interactions forces. In vitro enzymatic activity assays indicated an increase in the protein activity due to the saponin-protein interaction. Circular dichroism shows that saponin changes the ß-galactosidase secondary structure, favoring its protein-substrate interaction. Besides, changes in protein microenvironment due to the presence of saponin was observed by fluorescence spectroscopy. Isothermal titration calorimetry analyses suggested that saponins increased the affinity of ß-galactosidase with the artificial substrate o-nitrophenyl-ß-galactoside. The increase in the enzyme activity by saponins, demonstrated here, is important to new products development in food, cosmetic, and pharmaceutical industries.
Asunto(s)
Kluyveromyces/enzimología , Saponinas/farmacología , beta-Galactosidasa/efectos de los fármacos , Calorimetría , Dicroismo Circular , Cinética , Nitrofenilgalactósidos/metabolismo , Corteza de la Planta/química , Estructura Secundaria de Proteína , Quillaja/química , Espectrometría de Fluorescencia , beta-Galactosidasa/metabolismoRESUMEN
Nanostructured lipid carriers (NLCs) are a type of colloidal delivery system that was developed in the pharmaceutical industry to combine the advantages and eliminate the shortcomings of oil-in-water (O/W) nanoemulsions and solid lipid nanoparticles (SLNs). The hydrophobic core of the particles within NLCs consists of a solidified fat phase with a partially disorganized structure, which inhibits morphological changes and bioactive expulsion. In the present study, we formulated NLCs using a hot-homogenization approach using fully hydrogenated soybean oil (HSO) as the lipid phase and quillaja saponins as a natural surfactant. The NLCs formed had a low viscosity and milky white appearance similar to that of O/W nanoemulsions. The fabrication conditions were optimized, including the number of passes through the microfluidizer, stirring conditions, cooling rate, and emulsifier level. Unlike bulk HSO, the emulsified form had to be supercooled substantially to promote crystallization of the lipid droplets, which was attributed to differences in nucleation behavior. The crystallization temperature decreased with increasing saponin concentration, which was probably because smaller droplets were formed at higher emulsifier levels. For instance, at 3, 6, 9, and 12â¯wt% saponin, the degree of supercooling was 10, 15, 18, and 18⯰C, while the mean particle diameter was 0.82, 0.53, 0.41, and 0.44⯵m, respectively. The melting and crystallization behavior of the NLCs was characterized using an optical microscope and differential scanning calorimetry (DSC), while the morphology of the NLCs was characterized using transmission electron microscopy (TEM). This analysis showed that the NLCs contained spherical particles with a crystallization temperature around 31⯰C. This information may be useful for formulating NLC from natural ingredients for application in the food and beverage industry.
Asunto(s)
Emulsionantes/química , Emulsiones/química , Lípidos/química , Nanoestructuras/química , Saponinas de Quillaja/química , Coloides/química , Cristalización , Portadores de Fármacos/química , Microscopía Electrónica de Transmisión , Nanopartículas/química , Tamaño de la Partícula , Quillaja/química , Aceite de Soja/química , Tensoactivos , TemperaturaRESUMEN
High-order multiple emulsions are of great interest in both fundamental research and industrial applications as vehicles for their encapsulation capability of actives. In this work, we report a hierarchically multicompartmental highly stable triple emulsion by emulsifying and assembling of natural Quillaja saponin. Water-in-oil-in-(oil-in-water) (W2/O2/(O1/W1)) triple emulsion indicates that the compartmented system consisted of surfaced saponin-coated nanodroplets (SNDs) and dispersed oil globules, which in turn contained smaller aqueous droplets. The effects of formulation parameters, including lipophilic emulsifier content, oil fraction, and SND concentration, on the formation of multiple emulsions were systematically investigated. The assembly into fibrillar network of SNDs at the outer oil-water interface effectively protected the triple emulsion droplets against flocculation and coalescence, and strongly prevented the osmotic-driven water diffusion between the internal water droplets and the external water phase, thus contributing to superior stability during 180 days storage. All of these characteristics make the multicompartmentalized emulsions suitable to co-encapsulate a hydrophilic bioactive (gardenia blue) and two hydrophobic bioactives (eapsanthin and curcumin) in a single emulsion droplet hierarchically for the segregation and protection of multiple cargos. This approach offers a promising route toward accessing the next generation of functional deliveries and encapsulation strategies.
Asunto(s)
Curcumina/química , Sistemas de Liberación de Medicamentos/métodos , Extractos Vegetales/química , Quillaja/química , Saponinas/química , Composición de Medicamentos , Sistemas de Liberación de Medicamentos/instrumentación , Emulsionantes/química , Emulsiones/química , Glucósidos/química , Aceites/química , Tamaño de la Partícula , Agua/químicaRESUMEN
Delivery systems with multicompartmental structures that allow simultaneous delivery of several cargos are of great interest in both fundamental research and industrial applications. Here, we report a facile and easily scalable approach to fabricate multi-compartmentalized microdroplets for achieving programmed release of hydrophobic cargoes. Well-dispersed nanodroplets stabilized by natural Quillaja saponin served as an effective colloid stabilizer for fabricating microscale emulsion droplets with multicompartment architectures comprising many nanoscale droplets as a shell and single microscale core. Control of the number of nanodroplets allows accurate manipulation of the interface permeability for flexible and controllable release of volatile compounds (e.g., 2,3-butanedione, cis-3-hexen-1-ol, ethyl butyrate, d-limonene). More interestingly, the multicompartment microdroplets exhibited a higher flexibility for programmed release of different volatile compounds, as well as curcumin, during in vitro digestion by introducing cargos into the shell subcompartments or core microcompartment. The promising results highlight the power of this multi-compartmentalized system toward accessing a powerful platform for functional cargo delivery strategies.
Asunto(s)
Curcumina/química , Sistemas de Liberación de Medicamentos/métodos , Extractos Vegetales/química , Quillaja/química , Saponinas/química , Digestión , Sistemas de Liberación de Medicamentos/instrumentación , Emulsiones/química , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Tamaño de la PartículaRESUMEN
Immunoadjuvant Quillaja spp. tree saponins stimulate both cellular and humoral responses, significantly widening vaccine target pathogen spectra. Host toxicity of specific saponins, fractions and extracts may be rather low and further reduced using lipid-based delivery systems. Saponins contain a hydrophobic central aglycone decorated with several sugar residues, posing a challenge for viable chemical synthesis. These, however, may provide simpler analogs. Saponin chemistry affords characteristic interactions with cell membranes, which are essential for its mechanism of action. Natural sources include Quillaja saponaria barks and, more recently, Quillaja brasiliensis leaves. Sustainable large-scale supply can use young plants grown in clonal gardens and elicitation treatments. Quillaja genomic studies will most likely buttress future synthetic biology-based saponin production efforts.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Saponinas de Quillaja/farmacología , Quillaja/química , Adyuvantes Inmunológicos/síntesis química , Adyuvantes Inmunológicos/aislamiento & purificación , Animales , Humanos , Inmunidad Celular/efectos de los fármacos , Inmunidad Humoral/efectos de los fármacos , Estructura Molecular , Hojas de la Planta/química , Saponinas de Quillaja/síntesis química , Saponinas de Quillaja/aislamiento & purificación , Relación Estructura-ActividadRESUMEN
Saponins form a group of plant-produced glycosides with potential as biopesticide ingredients. The environmental fate of saponins has never been fully investigated. In the present study, we use QS-18, a specific saponin from Quillaja saponaria as an example, to quantify hydrolysis under different conditions of pH, temperature and water chemical composition. Saponin hydrolysis in buffer solutions was base-catalyzed and followed first-order kinetics. Thus, hydrolysis was slow at pH 5.1 with a half-life of 330 ± 220 d (26 °C), which increases to 0.06 ± 0.01 d at pH 10.0. Hydrolysis rates were highly sensitive to temperature with an activation energy of 56.9 ± 14.2 kJ mol-1 at pH 7.2. In strong contrast, hydrolysis in lake waters (pH 6.4-8.2) produced different patterns with a fast initial dissipation of 25 to 60% of the added saponin within the first five hours, followed by an extremely slow reaction with 25 to 75% unreacted saponin left after reaction times longer than 120 h. The fast dissipation followed by slow hydrolysis in lake water was hypothesized to be attributed to sorption and/or flocculation of saponins by inorganic nanoparticles and/or solutes in the lake water followed by inactivation of hydrolysis due to the sorption/flocculation. The present study demonstrates that saponins may hydrolyze slowly under acidic and cold conditions. In addition, it demonstrates that dissipation kinetics in natural waters may deviate substantially from the kinetics predicted based on laboratory experiments with "clean" buffered solutions. This emphasizes the need for a deeper understanding of the processes affecting the dissipation kinetics of potential toxins under natural conditions, as fate models based on laboratory derived kinetic data may be seriously flawed.
Asunto(s)
Agentes de Control Biológico/química , Lagos/química , Quillaja/química , Saponinas/química , Floculación , Concentración de Iones de Hidrógeno , Hidrólisis , Cinética , Nanopartículas/química , Soluciones , TemperaturaRESUMEN
Thymol oil-in-water nanoemulsions as a potential natural alternative for synthetic antioxidant agents were developed. The nanoemulsions were formulated using Quillaja Saponin bio-surfactant and green solvents including high oleic sunflower oil (HOSO), tricaprylin (TC), and cinnamaldehyde (CA). The 4% thymol nanoemulsions containing TC and HOSO remained stable during long-term storage (at least 30â¯d). The antioxidant activity (AA) of free thymol and thymol nanoemulsions was compared with butylated hydroxytoluene (BHT) and ascorbic acid. The results obtained from DPPH, FRAP, and CUPRAC antioxidant assays showed a substantial improvement (pâ¯<â¯0.05) of the AA of free thymol through emulsification. The outcomes from the AA of the nanoemulsions in raw chicken breast meat measured by the TBARS assay revealed a significant improvement (pâ¯<â¯0.05) of the AA when thymol was encapsulated. These nanoemulsions may be applicable in the food industry as well as in cosmetic and health care products.
Asunto(s)
Antioxidantes/química , Emulsiones/química , Nanoestructuras/química , Saponinas de Quillaja/química , Tensoactivos/química , Timol/química , Animales , Rastreo Diferencial de Calorimetría , Pollos , Carne/análisis , Tamaño de la Partícula , Quillaja/química , Quillaja/metabolismo , SonicaciónRESUMEN
BACKGROUND: Quillaja saponaria Mol. bark contains a high concentration of triterpene saponins that have been used for centuries as a cleansing, antiinflammatory and analgesic agent in Chilean folk medicine. In earlier studies, in mice, both the anti-inflammatory as well as the antinociceptive effect of the major sapogenin, quillaic acid have been demonstrated (QA). OBJECTIVE: To determine the antihyperalgesic effect of QA one and seven days after itpl administration of complete Freund's adjuvant (CFA) in male mice using the hot plate test in the presence of complete Freund's adjuvant (HP/CFA) as an acute and chronic skeletal muscle pain model. METHODS: The present study evaluated the antihyperalgesic activity of QA against acute and chronic skeletal muscle pain models in mice using the hot plate test in the presence of complete Freund's adjuvant (HP/CFA), at 24 h (acute assay) and 7 days (chronic assay) , with dexketoprofen (DEX) as the reference drug. RESULTS: In acute and chronic skeletal muscle pain assays, QA at 30 mg/kg ip elicited its maximal antihyperalgesic effects (65.0% and 53.4%) at 24 h and 7 days, respectively. The maximal effect of DEX (99.0 and 94.1 at 24 h and 7 days, respectively) was induced at 100 mg/kg. CONCLUSION: QA and DEX elicit dose-dependent antihyperalgesic effects against acute and chronic skeletal muscle pain, but QA is more potent than DEX in the early and late periods of inflammatory pain induced by CFA.