RESUMO
Lignin is endowed with antioxidant activity due to its diverse chemical structure. It is necessary to explore the relationship between antioxidant activity and the chemical structure of the lignin to develop its high-value utilization. Herein, we employed maleic acid (MA) as a hydrotropic agent to preferably isolate the lignin from distinct herbaceous sources (wheat straw and switchgrass) under atmospheric pressure conditions. The resultant acid hydrotropic lignin (AHL) isolated from wheat straw exhibited high radical scavenging rates, up to 98% toward DPPH and 94% toward ABTS. Further investigations indicated that during the MA hydrotropic fractionation (MAHF) process, lignin was carboxylated by MA at γ-OH of the side-chain, providing additional antioxidant activity from the carboxy group. It was also found that the radical scavenging rate of AHL has a positive correlation with carboxyl, phenolic hydroxyl contents, and the S-G (syringyl-guaiacyl) ratio, which could be realized by increasing the MAHF severity. Overall, this work underlies the enhancement origin of the antioxidant property of lignin, which will facilitate its application in biological fields as an efficient, cheap, and renewable antioxidant additive.
Assuntos
Antioxidantes , Biomassa , Fracionamento Químico , Lignina , Maleatos , Triticum , Lignina/química , Lignina/isolamento & purificação , Antioxidantes/química , Antioxidantes/farmacologia , Antioxidantes/isolamento & purificação , Triticum/química , Fracionamento Químico/métodos , Maleatos/química , Sequestradores de Radicais Livres/química , Sequestradores de Radicais Livres/farmacologia , Sequestradores de Radicais Livres/isolamento & purificação , Panicum/químicaRESUMO
Torsemide is a long acting pyridine sulfonylurea diuretic. Torsemide hydrochloride is widely used now, there are only a few organic acid salts reported. Cocrystallization with organic acids is an effective way to improve its solubility. Here, we reported maleate and phthalate of torsemide, in which the organic acid lost a proton transferring to the pyridine of torsemide, and torsemide interacted with organic acid through N+ - Hâ¯O- hydrogen bond to form salts crystal. Surprisingly, maleate showed a clear "spring" pattern in apparent solubility, whereas phthalate had a "spring-parachute" effect. Both crystalline salts kept a higher solubility than torsemide without falling. The "spring-parachute" effect of crystalline salts promoted rapid dissolution of torsemide and kept a high concentration, thereby increasing its bioavailability.
Assuntos
Cristalização , Sais , Solubilidade , Torasemida , Torasemida/química , Cristalização/métodos , Sais/química , Ligação de Hidrogênio , Diuréticos/química , Maleatos/química , Disponibilidade BiológicaRESUMO
3D bioprinting, a significant advancement in biofabrication, is renowned for its precision in creating tissue constructs. Collagen, despite being a gold standard biomaterial, faces challenges in bioink formulations due to its unique physicochemical properties. This study introduces a novel, neutral-soluble, photocrosslinkable collagen maleate (ColME) that is ideal for 3D bioprinting. ColME was synthesized by chemically modifying bovine type I collagen with maleic anhydride, achieving a high substitution ratio that shifted the isoelectric point to enhance solubility in physiological pH environments. This modification was confirmed to preserve the collagen's triple-helix structure substantially. Bioprinting parameters for ColME were optimized, focusing on adjustments to the bioink concentration, extrusion pressure, nozzle speed, and temperature. Results demonstrated that lower temperatures and smaller nozzle sizes substantially improved the print quality of grid structures. Additionally, the application of intermittent photo-crosslinking facilitated the development of structurally robust 3D multilayered constructs, enabling the stable fabrication of complex tissues. Cell viability assays showed that encapsulated cells within the ColME matrix maintained high viability after printing. When compared to methacrylated gelatin, ColME exhibited superior mechanical strength, resistance to enzymatic digestion, and overall printability, positioning it as an outstanding bioink for the creation of durable, bioactive 3D tissues.
Assuntos
Bioimpressão , Maleatos , Impressão Tridimensional , Animais , Maleatos/química , Bovinos , Sobrevivência Celular/efeitos dos fármacos , Colágeno/química , Reagentes de Ligações Cruzadas/química , Processos Fotoquímicos , Engenharia Tecidual , Materiais Biocompatíveis/química , Materiais Biocompatíveis/síntese química , Tinta , Alicerces Teciduais/química , Humanos , Colágeno Tipo I/químicaRESUMO
There has been a significant volume of work investigating the design and synthesis of new crystalline multicomponent systems via examining complementary functional groups that can reliably interact through the formation of noncovalent bonds, such as hydrogen bonds (H-bonds). Crystalline multicomponent molecular adducts formed using this approach, such as cocrystals, salts, and eutectics, have emerged as drug product intermediates that can lead to effective drug property modifications. Recent advancement in the production for these multicomponent molecular adducts has moved from batch techniques that rely upon intensive solvent use to those that are solvent-free, continuous, and industry-ready, such as reactive extrusion. In this study, a novel eutectic system was found when processing albendazole and maleic acid at a 1:2 molar ratio and successfully prepared using mechanochemical methods including liquid-assisted grinding and hot-melt reactive extrusion. The produced eutectic was characterized to exhibit a 100 °C reduction in melting temperature and enhanced dissolution performance (>12-fold increase at 2 h point), when compared to the native drug compound. To remove handling of the eutectic as a formulation intermediate, an end-to-end continuous-manufacturing-ready process enables feeding of the raw parent reagents in their respective natural forms along with a chosen polymeric excipient, Eudragit EPO. The formation of the eutectic was confirmed to have taken place in situ in the presence of the polymer, with the reaction yield determined using a multivariate calibration model constructed by combining spectroscopic analysis with partial least-squares regression modeling. The ternary extrudates exhibited a dissolution profile similar to that of the 1:2 prepared eutectic, suggesting a physical distribution (or suspension) of the in situ synthesized eutectic contents within the polymeric matrix.
Assuntos
Polímeros , Solubilidade , Análise dos Mínimos Quadrados , Polímeros/química , Química Farmacêutica/métodos , Maleatos/química , Composição de Medicamentos/métodos , Temperatura Alta , Ligação de Hidrogênio , Tecnologia de Extrusão por Fusão a Quente/métodos , Cristalização/métodosRESUMO
This study introduces a new NMR-based methodology for identification (ID) and quantification (purity, strength) assays of widely used amino acids. A detailed analysis of four amino acids and their available salts was performed with both a high-field (600â¯MHz) and a benchtop (60â¯MHz) NMR instrument. To assess sensitivity constraints, samples for 1H NMR analysis were initially prepared using only 10â¯mg of analyte and 1â¯mg of maleic acid (MA) as an internal calibrant (IC) and secondary chemical shift reference. The characteristic dispersion of the peak patterns indicating the presence or absence of a counterion (mostly chloride) was conserved at both high and low-field strength instruments, showing that the underlying NMR spectroscopic parameters, i.e., chemical shifts and coupling constants, are independent of the magnetic field strength. However, as the verbal descriptions of 1H NMR spectra are challenging in the context of reference materials and pharmaceutical monographs, an alternative method for the identification (ID) of amino acids is proposed that uses 13C NMR patterns from multiplicity-edited HSQC (ed-HSQC), which are both compound-specific and straightforward to document. For ed-HSQC measurements, the sample amount was increased to 30â¯mg of the analyte and several acquisition parameters were tested, including t1 increments used in the pulse program, number of scans, and repetition time. Excellent congruence with deviations <0.1â¯ppm was achieved for the 13C chemical shifts from 1D 13C NMR spectra (150â¯MHz) vs. those extracted from ed-HSQC (15â¯MHz traces). Finally, all samples of amino acid candidate reference materials were quantified by 1H qNMR (abs-qHNMR) at both 600 and 60â¯MHz. At high field, both IC and relative quantitations were performed, however, with the low-field instrument, only the IC method was used. The results showed that the analyzed reference material candidates were generally highly pure compounds. To achieve adequately low levels of uncertainty for such high-purity materials, the sample amounts were increased to 100â¯mg of analytes and 10â¯mg of the IC and replicates were analyzed for selected amino acids.
Assuntos
Aminoácidos , Espectroscopia de Ressonância Magnética , Aminoácidos/análise , Aminoácidos/química , Espectroscopia de Ressonância Magnética/métodos , Padrões de Referência , Calibragem , Espectroscopia de Prótons por Ressonância Magnética/métodos , Maleatos/química , Maleatos/análiseRESUMO
PURPOSE: This two-treatment, four-period, double-blind, randomized controlled crossover trial assessed the ability of two denture adhesives, both applied with a thin nozzle in a continuous application pattern, to prevent food infiltration beneath partial dentures. METHODS: Participants with mandibular partial dentures and a history of food particle infiltration were enrolled. All participants used both an optimized calcium/zinc partial salt of polyvinyl methyl ether/maleic acid (PVM/MA) denture adhesive and a calcium/sodium partial salt of PVM/MA test denture adhesive, twice each, throughout four study periods, according to a randomly assigned sequence. At each visit, participants underwent two assessments: once with no denture adhesive (baseline) and once with denture adhesive, 1 hour after adhesive application. For each assessment, participants ate one-half of the top of a poppy seed muffin, and a dental professional counted the seeds retained on the denture and mucosa, which was the primary variable. The change-from-baseline comparison was made for each treatment separately using a paired t-test or Wilcoxon Signed Rank test depending on the normality of the data. A between-treatment comparison for the change from baseline was performed using a crossover ANCOVA with treatment and period as fixed effects and participant as a random effect. The baseline poppy seed count was used as a covariate. RESULTS: 30 participants were enrolled; 29 completed the trial. Both denture adhesives achieved statistically significantly fewer retained seeds versus baseline (P< 0.001). The calcium/zinc adhesive reduced the seed count from baseline by 85.9% (6.18 vs 0.86), and the calcium/ sodium adhesive reduced seed count by 76.6% (6.04 vs 1.43). Comparing the two denture adhesives, the reduction in seed count from baseline was statistically significantly greater for the calcium/zinc adhesive versus the calcium/sodium formulation (P= 0.008). CLINICAL SIGNIFICANCE: These results support the recommendation of denture adhesive use for the prevention of food infiltration beneath partial dentures, with optimized calcium/zinc denture adhesive showing the greatest prevention benefit.
Assuntos
Adesivos , Estudos Cross-Over , Humanos , Masculino , Feminino , Idoso , Método Duplo-Cego , Maleatos/química , Pessoa de Meia-Idade , Alimentos , Zinco , Prótese Parcial , PolietilenosRESUMO
Styrene-maleic acid (SMA) copolymer has received much attention for its excellent solubilization characteristics. In this work, SMA copolymer brush-based chromatographic stationary phases were exploited and developed for the first time. First, SMA copolymer brush was in situ grown on the surface of spherical silica via living/controlled reversible addition-fragmentation chain transfer (RAFT) polymerization method. Subsequently, as a proof-of-concept demonstration, the copolymer was esterified by diethylene glycol mono-2-ethylhexyl ether (DGME) and 2-(2-ethylhexyloxy) ethanol (EHOE), respectively. The obtained Sil-SMA-DGME and Sil-SMA-EHOE copolymer-brush chromatographic stationary phases were characterized by transmission electron microscopy, Fourier transform infrared spectrometer, X-ray photoelectron spectroscopy, and thermogravimetric analysis, respectively. The chromatographic retention mechanism indicated that both the two packed columns exhibited hydrophilic/reverse mixed-mode retention modes. The maximum column efficiency was up to 71,000 N/m. The chromatographic separation performance evaluation indicated that the novel kind of stationary phases had excellent separation capabilities for hydrophilic, hydrophobic compounds and phospholipid standards. In addition, by combination with mass spectrometry identification, the Sil-SMA-DGME column was further exploited for separation and identification of phospholipids in human lung cancer cells. Totally, 9 classes including 186 phospholipid species were successfully identified. The results demonstrated the promising application prospects of the novel kind of SMA copolymer-brush chromatographic stationary phases.
Assuntos
Maleatos , Dióxido de Silício , Maleatos/química , Dióxido de Silício/química , Humanos , Interações Hidrofóbicas e Hidrofílicas , Poliestirenos/química , Esterificação , Cromatografia Líquida de Alta Pressão/métodos , Polímeros/químicaRESUMO
Salts of weakly basic drugs can partially dissociate in formulations, to give basic drugs and counter acids. The aim of the present study was to clarify the effect of physicochemical properties on the basic drug-acid-polymer interactions and salt-polymer miscibility, and to explain the influence mechanism at the molecular level. Six maleate salts with different physicochemical properties were selected and PVA was used as the film forming material. The relationship between the physicochemical properties and the miscibility was presented with multiple linear regression analysis. The existence state of salts in formulations were determined by XRD and Raman imaging. The stability of salts was characterized by NMR and XPS. The intermolecular interactions were investigated by FTIR and NMR. The results showed that the salt-PVA miscibility was related to polar surface area of salts and Tg of free bases, which represented hydrogen bond interaction and solubility potential. The basic drug-acid-PVA intermolecular interactions determined the existence state and bonding pattern of the three molecules. Meanwhile, the decrease of the stability after formulation increased the number of free bases in orodispersible films, which in turn affected the miscibility with PVA. The study provided references for the rational design of PVA based orodispersible films.
Assuntos
Álcool de Polivinil , Solubilidade , Álcool de Polivinil/química , Administração Oral , Estabilidade de Medicamentos , Maleatos/química , Química Farmacêutica/métodos , Ligação de Hidrogênio , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios XRESUMO
Lignin, a natural pol2ymer with a complex structure that is difficult to separate, is prone to C-C bond condensation during the separation process. To reduce the condensation of lignin, here, a novel method is proposed for separating the components by using a combination of maleic acid (MA)/ozone (O3) to co-treat wheat straw. The removal of lignin, glucan, and xylan was 38.07 ± 0.2%, 31.44 ± 0.1%, and 71.98 ± 0.1%, respectively, under the conditions of ball-milling of wheat straw for 6 h, reaction temperature of 60 °C, and O3 holding time of 9 min. Lignin-rich solutions were collected to extract the dissolved lignin (DL) after washing the treated samples. The DL obtained under MA/O3 conditions had a carboxyl group (-COOH) content of 2.96 mmol/g. The carboxyl group of MA underwent esterification with the hydroxyl group (-OH) at the γ position of lignin and O3 reacted on the positions of the lignin side chain or the phenolic ring, resulting in a break in the side chain and the opening of the phenolic ring to introduce the carboxyl group. The 2D-HSQC-NMR results revealed that the phenolic ring-opening reaction of lignin in the presence of O3 was essentially free of ß-ß and ß-5 condensation bonds.
Assuntos
Biomassa , Lignina , Maleatos , Ozônio , Triticum , Lignina/química , Triticum/química , Ozônio/química , Maleatos/químicaRESUMO
Expanding the detection information of wearable smart devices in applications has practical implications for their use in daily life and healthcare. Damage and breakage caused by mechanical injuries and continuous use are unavoidable for polymer matrices so self-healing properties are expected to be conferred on flexible sensors to extend their life and durability. In addition, a good linearity of relative resistance change vs. strain (gauge factor, GF) facilitates the streamlined conversion of electrical signals to 3D information of human motion, whereas existing works on sensors neglect the quantitative analysis of signals. This letter reports a self-healable flexible electronic sensor based on hydrogen bonding and electrostatic interaction between maleic acid-grafted natural rubber (MNR), polyaniline (PANI), and phytic acid (PA). MNR is the flexible matrix and the template for aniline (ANI) polymerization, and PA acts as the dopant and crosslinking agent. The MNR-PANI-PA sensor shows easy self-healing at room temperature, enhanced mechanical behaviour (â¼2.5 MPa, 1000% strain), and excellent linearity (GF of 13.8 over 250% strain and GF of 32.0 over 250-100% strain). Due to the highly linear relationship between ΔR/R and bending angle, the electrical signals of human limb movement can output relevant information on bending angle and frequency. By constructing a sensing array, changes in the position and magnitude of applied pressure could also be detected in real-time. Based on these advantages, the MNR-PANI-PA composite sensor is expected to have potential applications in health monitoring, body motion detection, and electronic skins.
Assuntos
Compostos de Anilina , Elastômeros , Movimento , Dispositivos Eletrônicos Vestíveis , Humanos , Elastômeros/química , Movimento/fisiologia , Condutividade Elétrica , Pressão , Ácido Fítico/química , Borracha/química , Maleatos/químicaRESUMO
Structural studies require the production of target proteins in large quantities and with a high degree of purity. For membrane proteins, the bottleneck in determining their structure is the extraction of the target protein from the cell membranes. A detergent that improperly mimics the hydrophobic environment of the protein of interest can also significantly alter its structure. Recently, using lipodiscs with styrene-maleic acid (SMA), copolymers became a promising strategy for the purification of membrane proteins. Here, we describe in detail the one-step affinity purification of potassium ion channels solubilized in SMA and sample preparation for future structural studies.
Assuntos
Maleatos , Poliestirenos , Canais de Potássio , Maleatos/química , Canais de Potássio/química , Canais de Potássio/metabolismo , Poliestirenos/química , Cromatografia de Afinidade/métodos , Estireno/química , Polímeros/química , Detergentes/química , HumanosRESUMO
The histamine H1 receptor (H1R) plays a pivotal role in allergy initiation and undergoes the necessity of devising a high-throughput screening approach centered on H1R to screen novel ligands effectively. This study suggests a method employing styrene maleic acid (SMA) extraction and His-tag covalent bonding to immobilize H1R membrane proteins, minimizing the interference of nonspecific proteins interference while preserving native protein structure and maximizing target exposure. This approach was utilized to develop a novel material for high-throughput ligand screening and implemented in cell membrane chromatography (CMC). An H1R-His-SMALPs/CMC model was established and its chromatographic performance (selectivity, specificity and lifespan) validated, demonstrating a significant enhancement in lifespan compared to previous CMC models. Subsequently, this model facilitated high-throughput screening of H1R ligands in the compound library and preliminary activity verification of potential H1R antagonists. Identification of a novel H1R antagonist laid the foundation for further development in this area.
Assuntos
Ensaios de Triagem em Larga Escala , Maleatos , Receptores Histamínicos H1 , Ligantes , Maleatos/química , Ensaios de Triagem em Larga Escala/métodos , Receptores Histamínicos H1/química , Receptores Histamínicos H1/metabolismo , Humanos , Histidina/química , Animais , Proteínas Imobilizadas/química , Proteínas Imobilizadas/metabolismo , Células CHO , Proteínas de Membrana/química , Proteínas de Membrana/metabolismo , Antagonistas dos Receptores Histamínicos H1/química , Poliestirenos/química , Cricetulus , Oligopeptídeos/químicaRESUMO
The construction and modification of a Graphene Oxide (GO) membrane, incorporating polyvinyl alcohol (PVA) cross-linked with maleic acid (MA) and supported by a nylon membrane, have been successfully completed. Systematic variations in PVA and MA concentrations were conducted to achieve membranes with favorable characteristics, stability, and excellent desalination performance. Optimization studies utilizing the Central Composite Design (CCD) revealed that the most optimal desalination results were obtained with 10 mL of PVA (0.1 mg mL-1) and 0.9 M of MA (GO-MA0.9-PVA10/Nylon membrane). Experimental findings demonstrated that the inclusion of PVA and MA resulted in an increased interlayer distance of GO and enhanced membrane stability. The addition of PVA increases GO membrane hydrophilicity, while the addition of MA reduces membrane hydrophilicity. The GO-MA0.9-PVA10/Nylon membrane exhibited the highest desalination performance, boasting a rejection value exceeding >99.9% and a permeance of 18.76 kg m-2.h-1 under 1% NaCl feed at a temperature of 50 °C. This membrane demonstrated consistent desalination performance stability over an extended period of up to 70 h. Moreover, it exhibited durability through 8 cycles of 24-h usage with washing treatment. In conclusion, the GO-MA0.9-PVA10/Nylon membrane is strongly recommended for practical applications, outperforming other membrane options based on the comprehensive evaluation of its stability and desalination efficiency.
Assuntos
Grafite , Membranas Artificiais , Álcool de Polivinil , Cloreto de Sódio , Purificação da Água , Grafite/química , Álcool de Polivinil/química , Purificação da Água/métodos , Cloreto de Sódio/química , Filtração/métodos , Maleatos/química , Salinidade , Interações Hidrofóbicas e Hidrofílicas , Nylons/químicaRESUMO
Incorporating starch, which is a potential biodegradable substitute for petroleum-based polymers, into conventional polymers is challenging owing to limitations in processability and weak-performing resulting materials. Herein, corn starch/polyvinyl alcohol (PVA) blend films (starch: PVA ratio of 50:50) were prepared via the solvent casting method using glycerol as a plasticizer and with varying concentrations of maleic acid as the crosslinking agent. Fourier transform infrared spectroscopy revealed the molecular interactions of the maleic acid crosslinker with the polymeric network of starch and PVA through an ester linkage. The properties of the films were strongly dependent on the maleic acid concentration. An increasing maleic acid concentration imparted hydrophobicity to the film; therefore, water swelling was significantly reduced, and water resistance was enhanced. The film containing 20 wt% maleic acid exhibited excellent barrier properties, with the lowest oxygen and water vapor transmission rates of 0.5 ± 0.2 cc/m2â day and 232.3 ± 5.4 g/m2â day, respectively. Moreover, the mechanical properties of the film improved with increasing crosslinking. This study demonstrates that the addition of maleic acid leads to an improvement in the overall performance of starch/PVA blend films. Therefore, maleic acid-crosslinked films can be used as barrier materials in food packaging applications.
Assuntos
Embalagem de Alimentos , Maleatos , Álcool de Polivinil , Amido , Álcool de Polivinil/química , Amido/química , Maleatos/química , Embalagem de Alimentos/métodos , Reagentes de Ligações Cruzadas/química , Interações Hidrofóbicas e Hidrofílicas , Água/química , Espectroscopia de Infravermelho com Transformada de Fourier , Permeabilidade , Plastificantes/química , VaporRESUMO
In the present study, a novel environment friendly dry method for preparation of guar gum maleate (GGM) with varying degrees of substitution (DS; 0.02-1.04) was optimized. GGM with a maximum DS of 1.04 was successfully synthesized using guar gum (GG) and maleic anhydride (MA) in proportion of 1: 1 at 80 °C with 4 h of reaction time. The activation energy for the reaction was determined to be 36.91 ± 3.61 kJ mol-1 with pre-exponential factor of 1392 min-1. Esterification of GG was confirmed by FT-IR and 13C NMR. Analysis using size exclusion chromatography (SEC) indicated a decrease in weight average molecular weight (Mw) of the polymer with an increase in polydispersity index (PDI) due to esterification. In comparison with GG, GGM displayed increased hydrophobicity and reduced thermal stability, as analysed by differential scanning calorimetry (DSC). Rheological studies of GGM revealed that initial apparent viscosity decreased with increasing DS. For the first time, the study offered valuable insights on GGM synthesis under dry solvent-less reaction conditions enabling simpler and scalable synthesis process.
Assuntos
Galactanos , Maleatos , Mananas , Gomas Vegetais , Gomas Vegetais/química , Galactanos/química , Mananas/química , Cinética , Maleatos/química , Peso Molecular , Viscosidade , Esterificação , Reologia , Temperatura , Técnicas de Química Sintética , Interações Hidrofóbicas e HidrofílicasRESUMO
We report a novel utilization of a pH modifier as a disproportionation retardant in a tablet formulation. The drug molecule of interest has significant bioavailability challenges that require solubility enhancement. In addition to limited salt/cocrystal options, disproportionation of the potential salt(s) was identified as a substantial risk. Using a combination of Raman spectroscopy with chemometrics and quantitative X-ray diffraction in specially designed stress testing, we investigated the disproportionation phenomena. The learnings and insight drawn from crystallography drove the selection of the maleate form as the target API. Inspired by the fumarate form's unique stability and solubility characteristics, we used fumaric acid as the microenvironmental pH modulator. Proof-of-concept experiments with high-risk (HCl) and moderate-risk (maleate) scenarios confirmed the synergistic advantage of fumaric acid, which interacts with the freebase released by disproportionation to form a more soluble species. The resultant hemifumarate helps maintain the solubility at an elevated level. This work demonstrates an innovative technique to mediate the solubility drop during the "parachute" phase of drug absorption using compendial excipients, and this approach can potentially serve as an effective risk-mitigating strategy for salt disproportionation.
Assuntos
Química Farmacêutica , Composição de Medicamentos , Fumaratos , Solubilidade , Fumaratos/química , Concentração de Íons de Hidrogênio , Composição de Medicamentos/métodos , Química Farmacêutica/métodos , Análise Espectral Raman/métodos , Difração de Raios X/métodos , Comprimidos/química , Sais/química , Maleatos/química , Excipientes/química , Disponibilidade BiológicaRESUMO
Multidrug resistance (MDR) is a dominant challenge in cancer chemotherapy failure. The over-expression of breast cancer resistance protein (BCRP) in tumorous cells, along with its extensive substrate profile, is a leading cause of tumor MDR. Herein, on the basis of styrene maleic acid (SMA) polymer membrane protein stabilization strategy and surface plasmon resonance (SPR) biosensor, a novel high-throughput screening (HTS) system for BCRP inhibitors has been established. Firstly, LLC-PK1 and LLC-PK1/BCRP cell membranes were co-incubated with SMA polymers to construct SMA lipid particles (SMALPs). PK1-SMALPs were thus immobilized in channel 1 of the L1 chip as the reference channel, and BCRP-SMALPs were immobilized in channel 2 as the detection channel to establish the BCRP-SMALPs-SPR screening system. The methodological investigation demonstrated that the screening system was highly specific and stable. Three active compounds were screened out from 26 natural products and their affinity constants with BCRP were determined. The KD of xanthotoxin, bergapten, and naringenin were 5.14 µM, 4.57 µM, and 3.72 µM, respectively. The in vitro cell verification experiments demonstrated that xanthotoxin, bergapten, and naringenin all significantly increased the sensitivity of LLC-PK1/BCRP cells to mitoxantrone with possessing reversal BCRP-mediated MDR activity. Collectively, the developed BCRP-SMALPs-SPR screening system in this study has the advantages of rapidity, efficiency, and specificity, providing a novel strategy for the in-depth screening of BCRP inhibitors with less side effects and higher efficacy.
Assuntos
Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP , Maleatos , Proteínas de Neoplasias , Ressonância de Plasmônio de Superfície , Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/antagonistas & inibidores , Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/metabolismo , Ressonância de Plasmônio de Superfície/métodos , Proteínas de Neoplasias/antagonistas & inibidores , Proteínas de Neoplasias/metabolismo , Proteínas de Neoplasias/análise , Humanos , Maleatos/química , Maleatos/farmacologia , Animais , Ensaios de Triagem em Larga Escala/métodos , Suínos , Poliestirenos/química , Técnicas Biossensoriais/métodosRESUMO
Styrene-maleic acid (SMA) and similar amphiphilic copolymers are known to cut biological membranes into lipid nanoparticles/nanodiscs containing membrane proteins apparently in their relatively native membrane lipid environment. Our previous work demonstrated that membrane raft microdomains resist such disintegration by SMA. The use of SMA in studying membrane proteins is limited by its heterogeneity and the inability to prepare defined derivatives. In the present paper, we demonstrate that some amphiphilic peptides structurally mimicking SMA also similarly disintegrate cell membranes. In contrast to the previously used copolymers, the simple peptides are structurally homogeneous. We found that their membrane-disintegrating activity increases with their length (reaching optimum at 24 amino acids) and requires a basic primary structure, that is, (XXD)n, where X represents a hydrophobic amino acid (optimally phenylalanine), D aspartic acid, and n is the number of repeats of these triplets. These peptides may provide opportunities for various well-defined potentially useful modifications in the study of membrane protein biochemistry. Our present results confirm a specific character of membrane raft microdomains.
Assuntos
Proteínas de Membrana , Peptídeos , Animais , Humanos , Membrana Celular/metabolismo , Membrana Celular/química , Maleatos/química , Microdomínios da Membrana/metabolismo , Microdomínios da Membrana/química , Proteínas de Membrana/química , Proteínas de Membrana/metabolismo , Peptídeos/química , Poliestirenos/química , Linhagem CelularRESUMO
BACKGROUND: Hydroxyapatite and its derivatives have been used for a lot of applications. One of them is drug release studies. Due to its low adhesion strength and lack of the strength and durability required for load-carrying applications, there is a need to improve the properties of hydroxyapatite. For this aim, the most important factors are increasing pH sensitivity and preventing coagulation. Mixing it with multifunctional polymers is the best solution. OBJECTIVES: The main objectives are: 1- preparing poly(acrylamide-co-acrylic acid/maleic acid)- hydroxyapatite (PAm-co-PAA/PMA-HApt), 2- assessment of (PAm-co-PAA/PMA-HApt) and dox-loaded poly(acrylamide-co-acrylic acid/maleic acid) (Dox-(PAm-co-PAA/PMA-HApt)) composite hydrogels, and 3- elucidating the difference in behavior of drug release studies between hydroxyapatite (HApt) and poly(acrylamide-co-acrylic acid/maleic acid) composite hydrogels. METHODS: A composite of PAm-co-PAA/PMA-HApt was prepared by direct polymerization of acrylamide-co-acrylic acid/maleic acid in a suspension of HApt. The drug loading and release features of PAm-co-PAA/PMA-HApt and HApt were then investigated for doxorubicin (dox) release. Using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and thermogravimetric analysis (TG/DTA), this unique composite hydrogel has been physicochemically investigated. Also, a colorimetric assay was used to assess the in vitro biocompatible support and anticancer activity of HApt and the newly developed composite hydrogel XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide) assay. RESULTS: According to the results of drug release studies of this new material, it is pH sensitive, and PAm-co-PAA/PMA-HApt demonstrated a faster release than HApt at 37°C in the acidic solution of pH 4.5 than in the neutral solution of pH 7.4. The XTT assay outcomes also demonstrated the biocompatibility of PAm-co-PAA/PMA-HApt and HApt and the cytotoxic effect of dox-loaded PAm-co-PAA/PMA-HApt. CONCLUSION: It should be inferred that the drug release profile was improved at pH 4.5 by the newly produced pH-sensitive composite hydrogel.
Assuntos
Doxorrubicina , Liberação Controlada de Fármacos , Durapatita , Hidrogéis , Maleatos , Doxorrubicina/química , Doxorrubicina/farmacologia , Durapatita/química , Maleatos/química , Hidrogéis/química , Hidrogéis/síntese química , Portadores de Fármacos/química , Portadores de Fármacos/síntese química , Humanos , Resinas Acrílicas/química , Resinas Acrílicas/síntese química , Acrilamidas/química , Concentração de Íons de HidrogênioRESUMO
Extracellular vesicles (EVs) are nanosized biomolecular packages involved in intercellular communication. EVs are released by all cells, making them broadly applicable as therapeutic, diagnostic, and mechanistic components in (patho)physiology. Sample purity is critical for correctly attributing observed effects to EVs and for maximizing therapeutic and diagnostic performance. Lipoprotein contaminants represent a major challenge for sample purity. Lipoproteins are approximately six orders of magnitude more abundant in the blood circulation and overlap in size, shape, and density with EVs. This study represents the first example of an EV purification method based on the chemically-induced breakdown of lipoproteins. Specifically, a styrene-maleic acid (SMA) copolymer is used to selectively breakdown lipoproteins, enabling subsequent size-based separation of the breakdown products from plasma EVs. The use of the polymer followed by tangential flow filtration or size-exclusion chromatography results in improved EV yield, preservation of EV morphology, increased EV markers, and reduced contaminant markers. SMA-based EV purification enables improved fluorescent labeling, reduces interactions with macrophages, and enhances accuracy, sensitivity, and specificity to detect EV biomarkers, indicating benefits for various downstream applications. In conclusion, SMA is a simple and effective method to improve the purity and yield of plasma-derived EVs, which favorably impacts downstream applications.