RESUMEN

Normal skin is the first line of defense in the human body. A burn injury makes the skin susceptible to bacterial infection, thereby delaying wound healing and ultimately leading to sepsis. The chances of biofilm formation are high in burn wounds due to the presence of avascular necrotic tissue. The most common pathogen to cause burn infection and biofilm is Pseudomonas aeruginosa. The purpose of this study was to create a microemulsion (ME) formulation for topical application to treat bacterial burn infection. In the present study, tea tree oil was used as the oil phase, Tween 80 and transcutol were used as surfactants, and water served as the aqueous phase. Pseudo ternary phase diagrams were used to determine the design space. The ranges of components as suggested by the design were chosen, optimization of the microemulsion was performed, and in vitro drug release was assessed. Based on the characterization studies performed, it was found that the microemulsion were formulated properly, and the particle size obtained was within the desired microemulsion range of 10 to 300 nm. The I release study showed that the microemulsion followed an immediate release profile. The formulation was further tested based on its ability to inhibit biofilm formation and bacterial growth. The prepared microemulsion was capable of inhibiting biofilm formation.

Asunto(s)

Antibacterianos , Biopelículas , Quemaduras , Sistemas de Liberación de Medicamentos , Emulsiones , Pseudomonas aeruginosa , Biopelículas/efectos de los fármacos , Quemaduras/tratamiento farmacológico , Quemaduras/microbiología , Pseudomonas aeruginosa/efectos de los fármacos , Sistemas de Liberación de Medicamentos/métodos , Antibacterianos/administración & dosificación , Antibacterianos/farmacología , Tamaño de la Partícula , Liberación de Fármacos , Tensoactivos/química , Polisorbatos/química , Aceite de Árbol de Té/administración & dosificación , Aceite de Árbol de Té/química , Aceite de Árbol de Té/farmacología , Química Farmacéutica/métodos , Humanos

RESUMEN

The influence of adding surfactants on the performance of high-solid anaerobic digestion of horticultural waste was extensively investigated in batch systems. Adding Tween series and polyethylene glycol series non-ionic surfactants had positive effects on biogas production, resulting in 370.1 mL/g VS and 256.6 mL/g VS with Tween 60 and polyethylene glycol 300 at a surfactant-to-grass mass ratio of 0.20, while the biogas production of anaerobic digestion without surfactants was 107.54 mL/g VS. The optimal and economically feasible choice was adding Tween 20 at a ratio of 0.08 g/g grass in high-solid anaerobic digestion. A kinetics model reliably represented the relationship between surfactant concentration and biogas production. The mechanism of surfactants working on lignocellulose was investigated. The improvement in high-solid anaerobic digestion by adding surfactants was attributed to the interaction between lignocelluloses and surfactants and the extraction of biodegradable fractions from the porous structure. An economic analysis showed that adding Tween 20 was likely to make a profit and be more feasible than adding Tween 60 and polyethylene glycol 300. This study confirms the enhancement in biogas production from horticultural waste by adding non-ionic surfactants.

Asunto(s)

Biocombustibles , Lignina , Tensoactivos , Tensoactivos/química , Anaerobiosis , Lignina/química , Polisorbatos/química , Polietilenglicoles/química , Biodegradación Ambiental , Reactores Biológicos , Cinética

RESUMEN

Protein-surfactant interaction is a dynamic interplay of electrostatic and hydrophobic forces that ensues from the folding of a protein. We employ impedance spectroscopy (IS), a label-free method, to investigate the unfolding and refolding of human serum albumin (HSA), a globular plasma protein, in the presence of two surfactants: polysorbate-20 (Tween-20), a nonionic surfactant, and sodium dodecyl sulfate (SDS), an anionic surfactant. The equivalent electrical analog circuit was predicted from impedance spectra of HSA in an aqueous solution at physiological pH and room temperature, focusing on varying the concentration of codissolved surfactants. A change in the dielectric constant (ε') and ionic conductivity (κ) is observed by comparing the surfactant-treated protein samples to the bare surfactant solutions to assess the conformational changes induced by surfactants in HSA. Far-UV circular dichroism analysis revealed a decrease in α-helices and an increase in ß-sheets and random coils upon SDS addition, which were reversed by Tween-20. Dynamic light scattering supported the findings by measuring changes in the hydrodynamic diameter (dh) of HSA. Unfolding and refolding of HSA with surfactants were also observed through photoluminescence spectroscopy by examining the microenvironment surrounding the single tryptophan (W) within the protein, and the thermodynamic parameters were obtained using the modified Stern-Volmer equation. Our research explores the intriguing domain of protein-surfactant interactions, offering insights with promising applications across diverse biological processes and IS as a suitable alternative technique for investigating protein conformational changes by studying the electrical response of the samples.

Asunto(s)

Espectroscopía Dieléctrica , Tensoactivos , Humanos , Tensoactivos/química , Desplegamiento Proteico , Polisorbatos/química , Albúmina Sérica/química , Dodecil Sulfato de Sodio/química , Replegamiento Proteico/efectos de los fármacos , Dicroismo Circular , Pliegue de Proteína

RESUMEN

The clinical usage of docetaxel (DTX) is severely hindered by the dose-limiting neutropenia and peripheral neurotoxicity of polysorbate 80-solubilized DTX injection, and there are no alternative formulations until now. In this study, we developed a new liposomal formulation of DTX to reduce its toxicities, accompanying with the greatly improved antitumor activity. The DTX was encapsulated into liposomes in the form of hydrophilic glutathione (GSH)-conjugated prodrugs using a click drug loading method, which achieved a high encapsulation efficiency (∼95 %) and loading capacity (∼30 % wt). The resulting liposomal DTX-GSH provided a sustained and efficient DTX release (∼50 % within 48 h) in plasma, resulting in a greatly improved antitumor activities as compared with that of polysorbate 80-solubilized DTX injection in the subcutaneous and orthotopic 4T1 breast tumor bearing mice. Even large tumors > 500 mm3 could be effectively inhibited and shrunk after the administration of liposomal DTX-GSH. More importantly, the liposomal DTX-GSH significantly decreased the neutropenia and peripheral neurotoxicity as compared with that of polysorbate 80-solubilized DTX injection at the equivalent dose. These data suggested that the liposomal DTX-GSH might become a superior alternative formulation to the commercial DTX injection.

Asunto(s)

Antineoplásicos , Docetaxel , Glutatión , Liposomas , Ratones Endogámicos BALB C , Docetaxel/administración & dosificación , Docetaxel/farmacocinética , Docetaxel/farmacología , Docetaxel/química , Animales , Ratones , Glutatión/química , Femenino , Antineoplásicos/administración & dosificación , Antineoplásicos/farmacocinética , Antineoplásicos/farmacología , Antineoplásicos/química , Línea Celular Tumoral , Profármacos/administración & dosificación , Profármacos/química , Profármacos/farmacología , Taxoides/administración & dosificación , Taxoides/farmacología , Taxoides/farmacocinética , Taxoides/química , Polisorbatos/química , Neutropenia/inducido químicamente , Neutropenia/tratamiento farmacológico

RESUMEN

Protein therapeutics, particularly antibodies, depend on maintaining their native structures for optimal function. Hydrophobic interfaces, such as the air-water interface, can trigger protein aggregation and denaturation. While completely avoiding such interfacial exposures during manufacturing and storage is impractical, minimizing them is crucial for enhancing protein drug stability and extending shelf life. In the biologics industry, surfactants like polysorbates are commonly used as additives (excipients) to mitigate these undesirable interfacial exposures. However, polysorbates, the most prevalent choice, have recognized limitations in terms of polydispersity, purity, and stability, prompting the exploration of alternative excipients. The present study identifies poly(N-isopropylacrylamide)-poly(ethylene glycol) (PNIPAM-PEG) block copolymers as a promising alternative to polysorbates. Due to its stronger affinity for the air-water interface, PNIPAM-PEG significantly outperforms polysorbates in enhancing protein stability. This claim is supported by results from multiple tests. Accelerated dynamic light scattering (DLS) experiments demonstrate PNIPAM-PEG's exceptional efficacy in preserving IgG stability against surface-induced aggregation, surpassing conventional polysorbate excipients (Tween 80 and Tween 20) under high-temperature conditions. Additionally, circular dichroism (CD) spectroscopy results reveal conformational alterations associated with aggregation, with PNIPAM-PEG consistently demonstrates a greater protective effect by mitigating negative shifts at λ â‰… 220 nm, indicative of changes in secondary structure. Overall, this study positions PNIPAM-PEG as a promising excipient for antibody therapeutics, facilitating the development of more stable and effective biopharmaceuticals.

Asunto(s)

Resinas Acrílicas , Excipientes , Polietilenglicoles , Estabilidad Proteica , Polietilenglicoles/química , Excipientes/química , Resinas Acrílicas/química , Estabilidad de Medicamentos , Productos Biológicos/química , Inmunoglobulina G/química , Polisorbatos/química , Agregado de Proteínas

RESUMEN

Tolterodine tartrate (TOT) is a selective anti-muscarinic drug to treat urinary urgency and overactive urinary bladder (OAB) occurring in children, renal disease and elderly patients. Oral delivery is associated with several adverse effects. We addressed HSPiP and QbD (quality by design)-oriented TOT loaded cationic nanoemulsions for transdermal delivery. Hansen solubility parameters (HSP) screened excipients based on theoretical solubility whereas, QbD optimized cationic nanoemulsions (CNE-TOT-6). Formulation characteristic parameters were desirable to execute targeted in vitro drug release and ex vivo permeation profiles. In vitro hemolysis was conducted at varied concentrations whereas, histopathological study supported the safety aspect of CNE-TOT6. A comparative bioavailability was carried out in a rat model. Capmul PG8 (CAP), tween 80, and PEG 400 (polyethylene glycol 400) were screened based on HSP and experimental solubility data. QbD suggested optimized content of CAP, tween 80, and PEG 400 to achieve the lowest value of size (184 nm), maximum % entrapment efficiency (87.2 %), high zeta potential (+32.6 mV), optimum viscosity (47.19 cP), and high extrudability (96 %) as compared to its gel. High gel consistency slowed down the drug release and permeation flux as compared to CNE-TOT6 suspension. Hemocompatible CNE-TOT6 increased pharmacokinetic parameters as compared to the control and gel without causing skin toxicity after application. Thus, HSPiP and QbD oriented cationic nanoemulsions are promising carriers to treat overactive urinary bladder.

Asunto(s)

Administración Cutánea , Disponibilidad Biológica , Liberación de Fármacos , Emulsiones , Antagonistas Muscarínicos , Polietilenglicoles , Polisorbatos , Absorción Cutánea , Solubilidad , Tartrato de Tolterodina , Animales , Masculino , Tartrato de Tolterodina/administración & dosificación , Tartrato de Tolterodina/farmacocinética , Antagonistas Muscarínicos/administración & dosificación , Antagonistas Muscarínicos/farmacocinética , Antagonistas Muscarínicos/química , Antagonistas Muscarínicos/toxicidad , Polisorbatos/química , Polietilenglicoles/química , Ratas , Excipientes/química , Nanopartículas/química , Cationes/química , Caprilatos/química , Hemólisis/efectos de los fármacos , Ratas Wistar , Sistemas de Liberación de Medicamentos/métodos , Piel/metabolismo , Ratas Sprague-Dawley , Glicéridos , Polímeros , Glicoles de Propileno

RESUMEN

Therapeutic monoclonal antibody (mAb) products for intravenous (IV) administration generally require aseptic compounding with a commercially available diluent. When the administration site is located away from the preparation site, the prepared dosing solution may need to be transported in a vehicle. The impact of vehicle transportation on the product quality of mAbs needs to be evaluated to define safe handling and transportation conditions for dosing solutions. The design and execution of actual vehicle transportation studies require considerable resources and time. In this study, we systematically developed three different laboratory equipment-based methods that simulate vehicle transportation stresses: orbital shaker (OS), reciprocating shaker (RS), and vibration test system (VTS)-based simulation methods. We assessed their feasibility by comparing the impact on product quality caused by each simulated method with that caused by actual vehicle transportation. Without residual polysorbate 80 (PS80) in the mAb dosing solution, transportation via a cargo van led to a considerable increase in the subvisible particle counts and did not meet the compendial specifications for the light obscuration method. However, the presence of as low as 0.0004%w/v (4 ppm) PS80 in the dosing solution stabilized the mAb against vehicle transportation stresses and met the compendial specifications. Vehicle transportation of an IV bag with headspace resulted in negligible micro air bubbles and foaming in both PS80-free and PS80-containing mAb dosing solutions. These phenomena were found to be comparable to the VTS-based simulated method. However, the OS- and RS-based simulated methods formed significantly more micro air bubbles and foaming in an IV bag with headspace than either actual vehicle transportation or the VTS-based simulated method. Despite the higher interfacial stress (micro air bubbles and foaming) in the dosing solution created by the OS- and RS-based simulated methods, 0.0004%w/v (4 ppm) PS80 in the dosing solution was found to be sufficient to stabilize the mAb. The study shows that under appropriate simulated conditions, the OS-, RS-, and VTS-based simulated methods can be used as practical and meaningful models to assess the impact and risk of vehicle transportation on the quality of mAb dosing solutions.

Asunto(s)

Anticuerpos Monoclonales , Transportes , Anticuerpos Monoclonales/química , Anticuerpos Monoclonales/administración & dosificación , Transportes/métodos , Composición de Medicamentos/métodos , Estudios de Factibilidad , Polisorbatos/química , Administración Intravenosa

RESUMEN

The hydrolysis of polysorbate surfactants in large molecule drug product formulations caused by residual host cell proteins presents numerous stability concerns for pharmaceuticals. The fatty acids (FA) released by polysorbate hydrolysis can nucleate into particulates or challenge the conformational stability of the proteinaceous active pharmaceutical ingredient (API). The loss of intact polysorbate may also leave the Drug Product (DP) vulnerable to interfacial stresses. Polysorbate 20 and 80 are available in several different quality grades (Multi-compendial, Super Refined, Pure Lauric Acid (PLA)/Pure Oleic Acid (POA)). All variations of polysorbate as well as three alternative surfactants: Brij L23, Brij O20 and Poloxamer 188 were compared for their ability to protect against air-water interfacial stresses as well as their risk for developing particulates when in the presence of lipoprotein lipase (LPL) (Pseudomonas). Results show a meaningful difference in the timing and morphology of FA particle formation depending on the type of polysorbate used. All grades of polysorbate, while susceptible to hydrolysis, still offered sufficient protection to interfacial stresses, even when hydrolyzed to concentrations as low as 0.005 % (w/v). Alternative surfactants that lack an ester bond were resistant to lipase degradation and showed good protection against shaking stress.

Asunto(s)

Ácidos Grasos , Polisorbatos , Tensoactivos , Polisorbatos/química , Tensoactivos/química , Ácidos Grasos/química , Ácidos Grasos/metabolismo , Hidrólisis , Esterasas/metabolismo , Estabilidad de Medicamentos , Tamaño de la Partícula , Ácidos Láuricos/química

RESUMEN

The appearance of an extrudate formulation was monitored during hot-melt extrusion (HME) continuous manufacturing over 3 days. The formulation matrix consisted of a polymeric component, copovidone, and a low molecular weight surfactant, polysorbate 80. Based on studies prior to the continuous manufacturing, the desired appearance of the target extrudate is translucent. Although process parameters such as feed rate and screw speed were fixed during the continuous manufacturing, the extrudate appearance changed over time from turbid to translucent. For root-cause investigation, the extrudates were analyzed offline by differential scanning calorimetry (DSC) and advanced polymer chromatography (APC™). Although the polysorbate 80 content of both turbid and translucent extrudates was within target, the glass transition temperature of the turbid extrudate was 2 °C above expected value. The observed turbidity was traced to lot-to-lot variability of the polysorbate 80 used in the continuous manufacturing, where APC™ analysis revealed that the relative content of the low molecular weight component varied from 23% to 27% in correlation with the evolution from turbid to translucent extrudates. This work stresses the importance of taking feeding material variability into account during continuous manufacturing.

Asunto(s)

Rastreo Diferencial de Calorimetría , Polisorbatos , Tensoactivos , Polisorbatos/química , Tensoactivos/química , Pirrolidinas/química , Tecnología de Extrusión de Fusión en Caliente/métodos , Compuestos de Vinilo/química , Excipientes/química , Temperatura de Transición , Química Farmacéutica/métodos

RESUMEN

We applied computing-as-a-service to the unattended system-agnostic miscibility prediction of the pharmaceutical surfactants, Vitamin E TPGS and Tween 80, with Copovidone VA64 polymer at temperature relevant for the pharmaceutical hot melt extrusion process. The computations were performed in lieu of running exhaustive hot melt extrusion experiments to identify surfactant-polymer miscibility limits. The computing scheme involved a massively parallelized architecture for molecular dynamics and free energy perturbation from which binodal, spinodal, and mechanical mixture critical points were detected on molar Gibbs free energy profiles at 180 °C. We established tight agreement between the computed stability (miscibility) limits of 9.0 and 10.0 wt% vs. the experimental 7 and 9 wt% for the Vitamin E TPGS and Tween 80 systems, respectively, and identified different destabilizing mechanisms applicable to each system. This paradigm supports that computational stability prediction may serve as a physically meaningful, resource-efficient, and operationally sensible digital twin to experimental screening tests of pharmaceutical systems. This approach is also relevant to amorphous solid dispersion drug delivery systems, as it can identify critical stability points of active pharmaceutical ingredient/excipient mixtures.

Asunto(s)

Excipientes , Polisorbatos , Excipientes/química , Polisorbatos/química , Vitamina E/química , Tensoactivos/química , Pirrolidinas/química , Simulación de Dinámica Molecular , Termodinámica , Tecnología de Extrusión de Fusión en Caliente/métodos , Compuestos de Vinilo

RESUMEN

BACKGROUND: Cutaneous Leishmaniasis (CL) remains a significant public health concern, particularly in the tropical and subtropical regions. Present treatment options for CL such as Fluconazole (FLZ) face limitations, including low solubility and bioavailability. This study aimed to address these challenges by investigating the use of nano-emulsions (NEs) to enhance the efficacy of FLZ against Leishmania major(L.major). MATERIALS AND METHODS: FLZ-NEs were formulated with oleic acid, Tween-20, and ethanol using low-energy emulsification at various surfactant/co-surfactant ratios. Subsequently, a comprehensive analysis was conducted to assess the physicochemical characteristics of the samples. This analysis encompassed stability, zeta potential, pH, viscosity, refractive index, and droplet size. We then studied the anti-parasitic properties of these optimized FLZ-NEs both in vitro and in vivo. RESULTS: The selected nano-emulsion (NE) formulation (2 % oleic acid, 20 % Tween 20, 10 % ethyl alcohol) showcased desirable properties like small droplet size (10.51 ± 0.24 nm), low dispersity (0.19 ± 0.03), and zeta potential value (- 0.41 ± 0.17 mV), key for stability and targeted drug delivery. This optimal formulation translated into remarkable efficacy. In vitro, FLZ-NEs demonstrated a threefold increase in their ability to combat promastigotes and a remarkable thirtyfold increase in their ability to combat amastigotes. Additionally, they demonstrated a ninefold advantage in their ability to specifically target parasites within infected macrophages, thereby attacking the infection site. These promising in vitro results translated into improved outcomes in vivo. Compared to other chemicals studied, FLZ-NE-treated mice showed decreased disease severity, weight growth, and quicker ulcer healing. It was further supported by histopathological research, which showed reduced tissue damage linked to Leishmania infection. CONCLUSION: These findings show the potential of nanotechnology-based drug delivery in improving anti-leishmanial treatment.

Asunto(s)

Emulsiones , Fluconazol , Leishmania major , Leishmaniasis Cutánea , Ratones Endogámicos BALB C , Leishmania major/efectos de los fármacos , Animales , Leishmaniasis Cutánea/tratamiento farmacológico , Leishmaniasis Cutánea/parasitología , Ratones , Fluconazol/administración & dosificación , Fluconazol/farmacología , Femenino , Nanopartículas/química , Ácido Oléico/química , Antiprotozoarios/farmacología , Antiprotozoarios/administración & dosificación , Antiprotozoarios/química , Composición de Medicamentos , Polisorbatos/química , Tamaño de la Partícula , Sistemas de Liberación de Medicamentos/métodos , Tensoactivos/química , Administración Tópica

RESUMEN

The aim of this study was to develop eye-drops with cefuroxime (CEF) sodium or vancomycin (VAN) hydrochloride, antibiotics that are instable in water. Anhydrous self-emulsifying oils (SEO) are proposed as a carrier and antibiotics are suspended. In the contact with tear fluid, the formulation should transform into emulsion, with fast dissolution of an antibiotic. CEF or VAN (5% w/w) was suspended in SEO carriers prepared by dissolving surfactants (Tween 20 or Span 80 5% w/w) in Miglyol, castor oil, or olive oil. Formulations with or without sodium citrate (2% w/w) were compared. Six-months or 1-year stability tests were carried out at 40 °C. The content of CEF and VAN was evaluated using HPLC and the potency of the antibiotic was assessed with agar diffusion method. In contact with water, drug particles suspended in SEO dissolved rapidly and o/w emulsion was formed. After 1-year at 40 °C, the content of degradation products was at most 0.5% in CEF and 4.0% in VAN formulations. The agar diffusion assay has shown that CEF and VAN loaded into SEO retained its potency against the sensitive microorganisms comparable to an aqueous solution. Therefore, SEO can be used as a novel carrier for the active substances which may not require improved solubility or absorption but need to be protected from moisture. This is a formulation that can be produced on industrial scale, with no limitation of stability or drug concentration.

Asunto(s)

Antibacterianos , Estabilidad de Medicamentos , Emulsiones , Soluciones Oftálmicas , Antibacterianos/administración & dosificación , Antibacterianos/química , Antibacterianos/farmacocinética , Emulsiones/química , Soluciones Oftálmicas/química , Hidrólisis , Aceite de Ricino/química , Cefuroxima/química , Cefuroxima/administración & dosificación , Cefuroxima/farmacocinética , Vancomicina/química , Vancomicina/administración & dosificación , Tensoactivos/química , Química Farmacéutica/métodos , Suspensiones , Agua/química , Solubilidad , Polisorbatos/química , Aceite de Oliva/química , Hexosas/química , Portadores de Fármacos/química

RESUMEN

In recent years, there has been a growing interest in regulating lipid digestion through the construction of various interfacial structures. In the present work, a series of complex interfacial structures were designed by combining Tween 80 in the aqueous phase and lecithin in the oil phase at different concentration ratios. The emulsification properties, the roles in regulating lipid digestion, and the interfacial dilatational rheological properties of the composite emulsifying systems were characterized. The results showed that the combination of Tween 80 and lecithin at different ratios could effectively modulate the rate of lipid digestion. The polyoxyethylene chains of Tween 80 formed a network, that provided a spatial obstacle for the adsorption of bile salts and lipases. Thus, Tween 80 significantly delayed the lipid digestion. The introduction of lecithin gradually replaced Tween 80 molecules at the interface, thus providing space for the adsorption of bile salts and lipases. In addition, as the ratio of lecithin concentration to Tween 80 increased, lecithin gradually became the dominant factor in the interfacial properties. As a result, the rate of lipid digestion was accelerated. Therefore, by compounding different ratios of lecithin and Tween 80, a series of emulsions with different lipid digestion rates were obtained. This research provides a basis for rationally designing food emulsions according to specific needs.

Asunto(s)

Ácidos y Sales Biliares , Emulsiones , Lecitinas , Lipasa , Polisorbatos , Polisorbatos/química , Lecitinas/química , Adsorción , Lipasa/química , Lipasa/metabolismo , Ácidos y Sales Biliares/química , Reología , Digestión , Metabolismo de los Lípidos , Nanoestructuras/química , Lípidos/química , Agua/química

RESUMEN

None of transitional lipid-based drug delivery systems (LBDDS) includes compositions containing one lipid and one water-soluble surfactant that form stable microemulsions. The conversion of liquid LBDDS to solid LBDDS has been limited by low drug loading. Previously, we have developed drug solid microemulsions containing one lipid and TPGS (a water-soluble surfactant) that achieved high drug loading and remarkably increased oral bioavailability. This study aimed to test if binary lipid systems (BLS), composed of one lipid and one water-soluble surfactant that form stable self-emulsifying microemulsions, is not an exclusive but widely applicable type of LBDDS for other lipids and surfactants and evaluate the influences of chemical structures of lipids and surfactants on microemulsions and solid microemulsions. We systemically identified new BLS by using a library of lipids and surfactants. Propylene glycol diesters and glycerol triesters were favorable for forming stable microemulsions with Tween 80, Cremophor EL, or TPGS. To the best of our knowledge, this is the first report exploring and confirming that the BLS is a new addition to traditional LBDDS, provides a promising option for researchers, and has the potential to increase drug loading to facilitate the development of solid microemulsions.

Asunto(s)

Sistemas de Liberación de Medicamentos , Emulsiones , Lípidos , Polietilenglicoles , Polisorbatos , Solubilidad , Tensoactivos , Vitamina E , Agua , Tensoactivos/química , Sistemas de Liberación de Medicamentos/métodos , Lípidos/química , Polisorbatos/química , Polietilenglicoles/química , Agua/química , Vitamina E/química , Glicerol/análogos & derivados

RESUMEN

There is increasing pharmaceutical interest in deep eutectic solvents not only as a green alternative to organic solvents in drug manufacturing, but also as liquid formulation for drug delivery. The present work introduces a hydrophobic deep eutectic solvent (HDES) to the field of lipid-based formulations (LBF). Phase behavior of a mixture with 2:1 M ratio of decanoic- to dodecanoic acid was studied experimentally and described by thermodynamic modelling. Venetoclax was selected as a hydrophobic model drug and studied by atomistic molecular dynamics simulations of the mixtures. As a result, valuable molecular insights were gained into the interaction networks between the different components. Moreover, experimentally the HDES showed greatly enhanced drug solubilization compared to conventional glyceride-based vehicles, but aqueous dispersion behavior was limited. Hence surfactants were studied for their ability to improve aqueous dispersion and addition of Tween 80 resulted in lowest droplet sizes and high in vitro drug release. In conclusion, the combination of HDES with surfactant(s) provides a novel LBF with high pharmaceutical potential. However, the components must be finely balanced to keep the integrity of the solubilizing HDES, while enabling sufficient dispersion and drug release.

Asunto(s)

Composición de Medicamentos , Liberación de Fármacos , Interacciones Hidrofóbicas e Hidrofílicas , Lípidos , Simulación de Dinámica Molecular , Solubilidad , Solventes , Tensoactivos , Solventes/química , Tensoactivos/química , Lípidos/química , Composición de Medicamentos/métodos , Polisorbatos/química , Ácidos Láuricos/química , Química Farmacéutica/métodos , Sulfonamidas/química , Sulfonamidas/administración & dosificación , Aceites/química

RESUMEN

PURPOSE: Polysorbates are among the most used surfactants in biopharmaceutical products containing proteins. Our work aims to develop a high-throughput fluorometric assay to further diversify the analytical toolbox for quantification of PSs. METHOD: The assay leverages the micelle activated fluorescence signal from N-Phenyl-1-Naphthylamine (NPN). The development and optimization of assay parameters were guided by the pre-defined analytical target profile. Furthermore, NMR was used to probe the interaction between protein, PS80 and NPN in the measurement system and understand protein interference. RESULTS: All assay parameters including excitation and emission wavelengths, standard curve, NPN concentration, and incubation time have been optimized and adapted to a microplate format, making it compatible with automated solutions that will be pursued in the near future to drive consistency and efficiency in our workflows. The specificity, accuracy, and precision of the assay have been demonstrated through a case study. Furthermore, NMR results provided additional insight into the change of the interaction dynamics between PS80 and NPN as the protein concentration increases. The results indicate minimal interaction between the protein and PS80 at lower concentration. However, when the concentration exceeds 75 mg/mL, there is a significant interaction between the protein and PS-80 micelle and monomer. CONCLUSION: A high-throughput fluorometric assay has been developed for quantification of polysorbates in biopharmaceutical samples including in-process samples, drug substance and drug product. The assay reported herein could serve as a powerful analytical tool for polysorbate quantification and control, complementing the widely used liquid chromatography with charged aerosol detection method.

Asunto(s)

Colorantes Fluorescentes , Fluorometría , Ensayos Analíticos de Alto Rendimiento , Micelas , Polisorbatos , Polisorbatos/química , Polisorbatos/análisis , Colorantes Fluorescentes/química , Ensayos Analíticos de Alto Rendimiento/métodos , Fluorometría/métodos , Tensoactivos/química , Tensoactivos/análisis , 1-Naftilamina/análogos & derivados , 1-Naftilamina/química , Productos Biológicos/análisis , Productos Biológicos/química , Espectroscopía de Resonancia Magnética/métodos

RESUMEN

Monoclonal antibodies (mAb) and other biological drugs are affected by enzymatic polysorbate (PS) degradation that reduces product stability and jeopardizes the supply of innovative medicines. PS represents a critical surfactant stabilizing the active pharmaceutical ingredients, which are produced by recombinant Chinese hamster ovary (CHO) cell lines. While the list of potential PS-degrading CHO host cell proteins (HCPs) has grown over the years, tangible data on industrially relevant HCPs are still scarce. By means of a highly sensitive liquid chromatography-tandem mass spectrometry method, we investigated seven different mAb products, resulting in the identification of 12 potentially PS-degrading hydrolases, including the strongly PS-degrading lipoprotein lipase (LPL). Using an LPL knockout CHO host cell line, we were able to stably overexpress and purify the remaining candidate hydrolases through orthogonal affinity chromatography methods, enabling their detailed functional characterization. Applying a PS degradation assay, we found nine mostly secreted, PS-active hydrolases with varying hydrolytic activity. All active hydrolases showed a serine-histidine-aspartate/glutamate catalytical triad. Further, we subjected the active hydrolases to pH-screenings and revealed a diverse range of activity optima, which can facilitate the identification of residual hydrolases during bioprocess development. Ultimately, we compiled our dataset in a risk matrix identifying PAF-AH, LIPA, PPT1, and LPLA2 as highly critical hydrolases based on their cellular expression, detection in purified antibodies, active secretion, and PS degradation activity. With this work, we pave the way toward a comprehensive functional characterization of PS-degrading hydrolases and provide a basis for a future reduction of PS degradation in biopharmaceutical drug products.

Asunto(s)

Anticuerpos Monoclonales , Cricetulus , Hidrolasas , Células CHO , Animales , Anticuerpos Monoclonales/química , Hidrolasas/metabolismo , Polisorbatos/química , Productos Biológicos/metabolismo , Humanos

RESUMEN

Plant-based meat analogs are being developed to address environmental, sustainability, health, and animal welfare concerns associated with real meat products. However, it is challenging to mimic the desirable physicochemical, functional, and sensory properties of real meat products using plant-based ingredients. Emulsion gels consisting of lipid droplets embedded in biopolymer matrices are commonly used to create products with appearances, textures, and sensory attributes like meat products. In this study, the impact of soybean oil droplet characteristics (concentration, size, and charge) on the physicochemical properties of potato protein gels was studied. The oil droplets were either coated by a non-ionic surfactant (Tween 20) or a plant protein (patatin) to obtain different surface properties. The introduction of the oil droplets caused the protein gels to change from mauve to off-white, which was attributed to increased light scattering. Increasing the oil droplet concentration in the emulsion gels decreased their shear modulus and Young's modulus, which was mainly attributed to the fact that the oil droplets were less rigid than the surrounding protein network. Moreover, increasing the oil droplet size made this effect more pronounced, which was attributed to their greater deformability. Competitive adsorption of proteins and surfactants at the oi-water interface in the Tween emulsion promoted emulsion instability. This research highlights the complexity of the interactions between oil droplets and protein networks in emulsion gels. These insights are important for the utilization of emulsion gels in the formulation of plant-based foods with improved quality attributes.

Asunto(s)

Emulsiones , Geles , Gotas Lipídicas , Proteínas de Plantas , Reología , Emulsiones/química , Geles/química , Gotas Lipídicas/química , Proteínas de Plantas/química , Tamaño de la Partícula , Aceite de Soja/química , Propiedades de Superficie , Productos de la Carne/análisis , Solanum tuberosum/química , Tensoactivos/química , Polisorbatos/química

RESUMEN

Octacosanol has various biological effects such as antioxidant, hypolipidemic and anti-fatigue. However, poor solubility has limited the application of octacosanol in food. The aim of this study was to prepare octacosanol nanoemulsions with better solubility, stability and safety and to investigate in vivo anti-fatigue effect. The food-grade formulation of the octacosanol nanoemulsions consisted of octacosanol, olive oil, Tween 80, glycerol and water with 0.1 %, 1.67 %, 23.75 %, 7.92 % and 66.65 % (w/w), respectively. The nanoemulsions had an average particle size of 12.26 ± 0.76 nm and polydispersity index of 0.164 ± 0.12, and showed good stability under different pH, cold, heat, ionic stress and long-term storage conditions. The results of animal experiments showed that the octacosanol nanoemulsions significantly prolonged the fatigue tolerance time, alleviated the fatigue-related biochemical indicators, and weakened the oxidative stress. Meanwhile, octacosanol nanoemulsions upregulated hepatic glycogen levels. Taken together, these findings suggested that octacosanol nanoemulsions have promising applications as anti-fatigue functional foods.

Asunto(s)

Emulsiones , Fatiga , Alcoholes Grasos , Emulsiones/química , Animales , Alcoholes Grasos/química , Alcoholes Grasos/farmacología , Fatiga/tratamiento farmacológico , Tamaño de la Partícula , Masculino , Agua/química , Estrés Oxidativo/efectos de los fármacos , Ratas , Antioxidantes/farmacología , Antioxidantes/química , Ratas Sprague-Dawley , Solubilidad , Hígado/efectos de los fármacos , Hígado/metabolismo , Glucógeno/metabolismo , Glucógeno/química , Polisorbatos/química , Polisorbatos/farmacología , Nanopartículas/química

RESUMEN

In this study, we aimed to develop a solid self-nanoemulsifying drug delivery system (S-SNEDDS) and a solid self-nanoemulsifying granule system (S-SNEGS) to enhance the solubility and oral bioavailability of celecoxib. This process involved the preparation of a liquid SNEDDS (L-SNEDDS) and its subsequent solidification into a S-SNEDDS and a S-SNEGS. The L-SNEDDS consisted of celecoxib (drug), Captex® 355 (Captex; oil), Tween® 80 (Tween 80; surfactant) and D-α-Tocopherol polyethylene glycol 1000 succinate (TPGS; cosurfactant) in a weight ratio of 3.5:25:60:15 to produce the smallest nanoemulsion droplet size. The S-SNEDDS and S-SNEGS were prepared with L-SNEDDS/Ca-silicate/Avicel PH 101 in a weight ratio of 103.5:50:0 using a spray dryer and 103.5:50:100 using a fluid bed granulator, respectively. We compared the two novel developed systems and celecoxib powder based on their solubility, dissolution rate, physicochemical properties, flow properties and oral bioavailability in rats. S-SNEGS showed a significant improvement in solubility and dissolution rate compared to S-SNEDDS and celecoxib powder. Both systems had been converted from crystalline drug to amorphous form. Furthermore, S-SNEGS exhibited a significantly reduced angle of repose, compressibility index and Hausner ratio than S-SNEDDS, suggesting that S-SNEGS was significantly superior in flow properties. Compared to S-SNEDDS and celecoxib powder, S-SNEGS increased the oral bioavailability (AUC value) in rats by 1.3 and 4.5-fold, respectively. Therefore, S-SNEGS wolud be recommended as a solid self-nanoemulsifying system suitable for poorly water-soluble celecoxib.

Asunto(s)

Disponibilidad Biológica , Celecoxib , Sistemas de Liberación de Medicamentos , Emulsiones , Ratas Sprague-Dawley , Solubilidad , Agua , Celecoxib/química , Celecoxib/farmacocinética , Celecoxib/administración & dosificación , Animales , Emulsiones/química , Administración Oral , Masculino , Agua/química , Ratas , Tamaño de la Partícula , Tensoactivos/química , Nanopartículas/química , Polisorbatos/química