RESUMO
Antimicrobial peptides have appeared to be promising candidates for therapeutic purposes due to their broad antimicrobial activity and non-toxicity. Histatin-5 (Hst-5) is a notable salivary antimicrobial peptide that exhibited therapeutic properties in the oral cavity. Oral mucositis is an acute inflammation of the oral cavity, following cancer therapy. The current treatment methods of oral mucositis have low effectiveness. The aim of this study was to design, formulate and characterize a mucoadhesive gel delivery system for Hst-5 usage in the treatment of oral mucositis. Carbopol 934 and hydroxypropyl methylcellulose (HPMC) have been used in the development of a Hst-5 mucoadhesive gel that was optimized by using Box-Behnken design. The optimized formulation was evaluated in-vitro, based on mucoadhesive strength, viscoelasticity, spreadability, release rate, peptide secondary structure analysis, antimicrobial activity, and storage stability. The efficacy of Hst-5 gel was assessed in vivo in a chemotherapy-induced mucositis model. The results showed a sustained release of Hst-5 from the new formulation. Hst-5 gel exerted antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Candida albicans. The histopathological, immunohistochemical and statistical analysis showed that the Hst-5 gel had wound healing activity in vivo. The findings of this study indicate that the mentioned compound possesses promising potential as a novel and efficient therapeutic agent in managing oral mucositis. Moreover, the results suggest that the compound is commercially feasible for further development and utilization.
Assuntos
Mucosite , Estomatite , Histatinas , Estomatite/tratamento farmacológico , Candida albicans , Escherichia coliRESUMO
The oral cavity is one of the most important routes for local and systemic drug delivery, as it has a large surface, high permeability, and rich blood supply. Oral mucosal drug delivery has some advantages, such as enhancing bioavailability, preventing first-pass metabolism, reducing dose frequency, and non-invasiveness. In recent years, notable oral mucoadhesive patents were introduced to the pharmaceutical field, which indicates promising potentials for therapeutic purposes. Oral mucosal drug delivery can play a key role to deliver the biological drugs, such as antimicrobial peptides. This article gives an overview of oral mucoadhesive drug delivery systems and provides basic principles for the researchers to overcome the problems associated with the formulation design.
Assuntos
Sistemas de Liberação de Medicamentos , Mucosa Bucal , Disponibilidade Biológica , Mucosa Bucal/metabolismo , Permeabilidade , Preparações Farmacêuticas/metabolismoRESUMO
High porous particles with specific aerodynamic properties were processed by the spray freeze-drying (SFD) method. Comprehensive knowledge about all aspects of the SFD method is required for particle engineering of various pharmaceutical products with good flow properties. In this review, different types of the SFD method, the most frequently employed excipients, properties of particles prepared by this method, and most recent approaches concerning SFD are summarized. Generally, this technique can prepare spherical-shaped particles with a highly porous interior structure, responsible for the very low density of powders. Increasing the solubility of spray freeze-dried formulations achieves the desired efficacy. Also, due to the high efficiency of SFD, by determining the different features of this method and optimizing the process by model-based studies, desirable results for various inhaled products can be achieved and significant progress can be made in the field of pulmonary drug delivery.
Assuntos
Química Farmacêutica , Administração por Inalação , Química Farmacêutica/métodos , Liofilização/métodos , Tamanho da Partícula , Pós/químicaRESUMO
BACKGROUND: Effect of nifedipine on pressure ulcer (PU) healing has not been evaluated in the human subjects yet. STUDY QUESTION: In this study, the effect of topical application of nifedipine 3% ointment on PU healing in critically ill patients was investigated. STUDY DESIGN: This was a randomized, double-blind, placebo-controlled clinical. MEASURES AND OUTCOMES: In this study, 200 patients with stage I or II PU according to 2-digit Stirling Pressure Ulcer Severity Scale were randomized to receive topical nifedipine 3% ointment or placebo twice daily for 14 days. Changes in the size and stage of the ulcers were considered as primary outcome of the study. The stage of the ulcers at baseline and on day 7 and day 14 of study was determined by using 2-digit stirling scale. In addition, the surface area of the wounds was estimated by multiplying width by length. RESULTS: In total, 83 patients in each group completed the study. The groups were matched for the baseline stage and size of PUs. Mean decrease in the stage of PU in the nifedipine group was significantly higher than the placebo group on day 7 (-1.71 vs. -0.16, respectively, P < 0.001) and day 14 (-0.78 vs. -0.09, respectively, P < 0.001). Furthermore, the mean decrease in the surface area of PU was significantly higher in the nifedipine group compared with the placebo group on day 7 (-1.44 vs. -0.32, respectively, P < 0.001) and day 14 (-2.51 vs. -0.24, respectively, P < 0.001) of study. CONCLUSIONS: Topical application of nifedipine 3% ointment for 14 days significantly improved the healing process of stage I or II PUs in critically ill patients.
Assuntos
Nifedipino , Úlcera por Pressão , Método Duplo-Cego , Humanos , Pomadas , Úlcera por Pressão/tratamento farmacológico , CicatrizaçãoRESUMO
Adapalene (ADA) is believed to be one of the topical treatments utilized commonly in case of acne. Nanostructured lipid carriers (NLCs) have been established as an effective carrier system with certain advantages, for instance increased solubility, drug targeting, controlled drug release, and stability of ADA. This study was conducted to obtain the formulation with a good therapeutic property. All formulations were formed by probe sonicator and its characterizations were analyzed. Finally, the therapeutic effects of 0.1% ADA-loaded nanostructured lipid carriers (NLC-ADA) were evaluated. This formulation had a great entrapment efficiency (EE) that illustrated a controlled drug release profile. A pilot clinical evaluation conducted on 15 patients (age 25.23 ± 12.24 years) with mild to moderate acne vulgaris lesions. The results demonstrated significant reduction in acne severity index and the number of inflammatory and noninflammatory lesions after 12 weeks of treatment (P-value .02, .04, and .01, respectively). Subjective results were confirmed with significant improvement in size and intensity of porphyrin production in pilosebaceous follicles (P-value = .03). The study demonstrated that the formulation was safe and revealed the proper improvement rate of acne lesions after 12 weeks.
Assuntos
Acne Vulgar , Nanoestruturas , Preparações Farmacêuticas , Acne Vulgar/diagnóstico , Acne Vulgar/tratamento farmacológico , Adapaleno , Adolescente , Adulto , Criança , Humanos , Lipídeos , Adulto JovemRESUMO
The great potential of hydroxypropyl beta-cyclodextrin (HPßCD), as a dried-protein stabilizer, has been attributed to various mechanisms namely water-replacement, vitrification and surfactant-like effects. Highlighting the best result in our previous study (weight ratio IgG: HPßCD of 1:0.4), herein we designed to evaluate the efficacy of upper (1:2) and lower (1:0.05) ratios of HPßCD in stabilization and aerosol properties of spray freeze-dried IgG. The protective effect of HPßCD, as measured by size exclusion chromatography (SEC-HPLC) was most pronounced at C3' and C3â³, IgG:trehalose:HPßCD ratios of 1:2:0.25 and 1:2:0.05 with aggregation rate constants of 0.46 ± 0.02 and 0.58 ± 0.01 (1/month), respectively. The secondary conformations were analyzed through Fourier transform infrared spectroscopy (FTIR) and all powders well-preserved with the lack of any visible fragments qualified through sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PPAGE). Scanning electron microscopy (SEM) and twin stage impinger (TSI) were employed to characterize the suitability of particles for further inhalation therapy of antibodies and the highest values of fine particle fraction (FPF) were achieved by C3' and C3â³, 56.43 and 48.12%. The powders produced at the current ratio 1:2:0.25 and 1:2:0.05 are superior to our previous examination with regards to manifesting lower aggregation and comparable FPF values.
Assuntos
2-Hidroxipropil-beta-Ciclodextrina/química , Química Farmacêutica , Excipientes/química , Imunoglobulina G/administração & dosagem , Administração por Inalação , Aerossóis , Cromatografia em Gel , Estabilidade de Medicamentos , Liofilização , Humanos , Imunoglobulina G/química , Tamanho da Partícula , Pós , Tensoativos/química , Trealose/químicaRESUMO
Erythropoietin (EPO), a hematopoietic factor, is one of the promising neuroprotective candidates in neurodegenerative disorders such as Alzheimer's disease (AD). Due to the high molecular weight, hydrophilicity and rapid clearance from circulation, EPO could not completely pass the blood-brain barrier in the case of systemic administration. To overcome this limitation, EPO-loaded Solid Lipid Nanoparticle (EPO-SLN) was developed in this study using a double emulsion solvent evaporation method (W1/O/W2). Glycerin monostearate (GMS), span®80/span®60, Dichloromethane (DCM) and tween®80 were chosen as lipid, internal phase surfactants, solvent, and external aqueous phase surfactant, respectively. After physicochemical evaluations, the effect of EPO-SLN on the beta-amyloid-induced AD-like animal model was investigated. In vivo evaluations, it was demonstrated that the memory was significantly restored in cognitive deficit rats treated with EPO-SLN compared to the rats treated with native drug using the Morris water maze test. In addition, EPO-SLN reduced the oxidative stress, ADP/ATP ratio, and beta-amyloid plaque deposition in the hippocampus more effectively than the free EPO. Hence, the designed SLN can be regarded as a promising system for safe and effective delivery of EPO in the AD.
Assuntos
Doença de Alzheimer/tratamento farmacológico , Eritropoetina/uso terapêutico , Aprendizagem em Labirinto/efeitos dos fármacos , Transtornos da Memória/tratamento farmacológico , Nanopartículas/uso terapêutico , Fármacos Neuroprotetores/uso terapêutico , Doença de Alzheimer/induzido quimicamente , Peptídeos beta-Amiloides , Animais , Modelos Animais de Doenças , Eritropoetina/administração & dosagem , Hipocampo/efeitos dos fármacos , Masculino , Nanopartículas/administração & dosagem , Fármacos Neuroprotetores/farmacologia , Ratos , Ratos Wistar , Resultado do TratamentoRESUMO
Colorectal cancer (CRC) is the third leading cause of cancer-related death worldwide. The prognosis and overall survival of CRC are known to be significantly correlated with the overexpression of PD-L1. Since combination therapies can significantly improve therapeutic efficacy, we constructed doxorubicin (DOX) conjugated and anti-PD-L1 targeting gold nanoparticles (PD-L1-AuNP-DOX) for the targeted chemo-photothermal therapy of CRC. DOX and anti-PD-L1 antibody were conjugated to the α-terminal end group of lipoic acid polyethylene glycol N-hydroxysuccinimide (LA-PEG-NHS) using an amide linkage, and PD-L1-AuNP-DOX was constructed by linking LA-PEG-DOX, LA-PEG-PD-L1, and a short PEG chain on the surface of AuNP using thiol-Au covalent bonds. Physicochemical characterizations and biological studies of PD-L1-AuNP-DOX were performed in the presence of near-infrared (NIR) irradiation (biologic studies were conducted using cellular uptake, apoptosis, and cell cycle assays in CT-26 cells). PD-L1-AuNP-DOX (40.0 ± 3.1 nm) was successfully constructed and facilitated the efficient intracellular uptake of DOX as evidenced by pronounced apoptotic effects (66.0%) in CT-26 cells. PD-L1-AuNP-DOX treatment plus NIR irradiation significantly and synergistically suppressed the in vitro proliferation of CT-26 cells by increasing apoptosis and cell cycle arrest. The study demonstrates that PD-L1-AuNP-DOX in combination with synergistic targeted chemo-photothermal therapy has a considerable potential for the treatment of localized CRC.
Assuntos
Anticorpos/uso terapêutico , Antígeno B7-H1/imunologia , Neoplasias Colorretais/tratamento farmacológico , Doxorrubicina/uso terapêutico , Ouro/química , Nanopartículas Metálicas/química , Fotoquimioterapia/métodos , Animais , Anticorpos/administração & dosagem , Apoptose/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Terapia Combinada/métodos , Sistemas de Liberação de Medicamentos/métodos , Camundongos , Polietilenoglicóis/química , Espécies Reativas de Oxigênio/metabolismo , Succinimidas/química , Ácido Tióctico/químicaRESUMO
Ciprofloxacin is a commonly prescribed antibiotic for treatment of pulmonary infections. Nanocarriers such as nanomicelles can increase the drug residence time in the lungs and enhance their antibacterial effects. Dry powder inhalers (DPIs) are the preferred pulmonary drug delivery system and preparation of an optimum nanoaggregate from nanomicelles by means of spray drying would be valuable. The two-level full factorial design was performed in 16 runs. The effects of carrier type, anti-adhesion agent type, carrier to nanoparticle ratio and anti-adhesion agent to carrier ratio on the size of the microparticles, their in vitro pulmonary deposition, and redispersibility were investigated. Its antibacterial effects against Pseudomonas aeruginosa, Klebsiella pneumoniae, and Streptococcus pneumoniae also were investigated. All independent variables were fitted into two-factorial interaction models. The optimum nanoaggregate was prepared using mannitol and L-phenylalanine with a D0.5 of 1.7 µm and 60% fine particles. The process had no negative effect on the stability or drug release profile of the nanomicelles. The antibacterial effects of ciprofloxacin against microorganisms increased significantly. This spray drying process could be used for preparation of an optimum DPI from polymeric nanomicelles. This formulation could increase the efficacy of ciprofloxacin for treatment of pulmonary infections.
Assuntos
Antibacterianos/administração & dosagem , Ciprofloxacina/administração & dosagem , Dessecação , Inaladores de Pó Seco , Excipientes/química , Micelas , Polímeros/química , Administração por Inalação , Antibacterianos/química , Antibacterianos/farmacologia , Infecções Bacterianas/tratamento farmacológico , Ciprofloxacina/química , Ciprofloxacina/farmacologia , Dessecação/métodos , Liberação Controlada de Fármacos , Humanos , Klebsiella pneumoniae/efeitos dos fármacos , Manitol/química , Nanopartículas/química , Tamanho da Partícula , Fenilalanina/química , Pós , Pseudomonas aeruginosa/efeitos dos fármacos , Streptococcus pneumoniae/efeitos dos fármacosRESUMO
PURPOSE: Spray-freeze drying (SFD) is a recently applied method to develop pharmaceutical powders. This study aimed to analyze the competence of Trehalose, Mannitol, Lactose, and Sorbitol instability and aerosolization of Immunoglobulin G (IgG) via SFD. METHODS: Induced soluble aggregates were quantified at 0 and 3 months, and 45 °C using size-exclusion chromatography. Conformation and thermogravimetric assessments were done by Fourier transform infrared spectroscopy and differential scanning calorimetry. Laser light scattering was performed to determine the particle sizes. Aerodynamic features were characterized by twin stage impinger and scanning electron microscopy. RESULTS: Although sugars/polyols preferably stabilized IgG following the process, storage stabilization was achieved in Trehalose, Trehalose-Lactose, Lactose, and Trehalose-Mannitol-based powders with soluble aggregates <5%. The conformation of antibody was preserved with ß sheet content from 66.28% to 76.37%. Particle sizes ranged from 5.23 to 8.12 µm. Mannitol exhibited the best aerodynamic behavior, fine particle fraction (FPF: 70%) but high degree of protein aggregation during storage. CONCLUSIONS: SFD could favorably stabilize antibody using Trehalose and its combination with Lactose and Mannitol, and also, Lactose alone. Sorbitol disturbed IgG powder recovery. Incorporation of other types of excipient is required for efficient respiratory delivery of IgG molecules.
Assuntos
Química Farmacêutica/métodos , Dissacarídeos/química , Imunoglobulina G/química , Tamanho da Partícula , Combinação de Medicamentos , Composição de Medicamentos , Estabilidade de Medicamentos , Excipientes/química , Liofilização/métodos , Manitol/química , Trealose/químicaRESUMO
The physical and structural stability of freeze-dried immunoglobulin G (IgG) were examined by applying trehalose and amino acids (glycine, phenylalanine, and serine). The efficacy of amino acids was statistically compared considering their side-chain characteristics. The amount of amino acids (X1) and trehalose (X2) was considered as independent variables. Size exclusion chromatography (SEC-HPLC) was utilized to calculate the soluble aggregates, as dependent variables. The amounts of excipients were optimized through the central composite design (CCD). The beta-sheet conformation of IgG was quantified by Fourier transform infrared spectroscopy (FTIR). Thermal behavior and molecular integrity of IgG were evaluated by differential scanning calorimetry (DSC) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Optimized formulations of powders were as follows: 24.5 mg serine-139.5 mg trehalose, 14 mg glycine-118 mg Trehalose, and 25 mg phenylalanine-139.5 mg trehalose. The amounts of soluble aggregates after processing were 0, 4.50, and 2.20%, respectively. The corresponding induced aggregates following storage conditions were 1.02, 7.0, and 3.70%. In all preparations, there were no detectable fragments. The native conformation of IgG was well preserved in the presence of amino acids. Excluding the glycine-based sample with minor endotherm at about 45°C, serine and phenylalanine incorporating powders were fully amorphous at examination temperatures. Trehalose was more potent than the amino acids in the stabilization of IgG. Serine was the most effective amino acid; phenylalanine and glycine were the next ones, respectively. Glycine crystallization was assumed to have accounted for low stabilization capability. The statistically synergistic phenomenon was only observed in the co-application of trehalose and phenylalanine. Graphical abstract.
Assuntos
Aminoácidos/química , Imunoglobulina G/química , Trealose/química , Composição de Medicamentos , Estabilidade de Medicamentos , Armazenamento de Medicamentos , Liofilização , Glicina/químicaRESUMO
The airways are verified as a relevant route to improve antibody therapeutic index with superior lung concentration but limited passage into systemic blood stream. The current research aimed to process spray-dried (SD) powder of Infliximab to assess the feasibility of respiratory delivery of antibody for local suppression of lung-secreted tumor necrosis factor α (TNFα). Molecular and structural stability of powders were determined through size exclusion chromatography (SEC-HPLC) and Fourier transform infrared (FTIR) spectroscopy. Particle properties were characterized by laser light scattering, twin stage impinger (TSI), and scanning electron microscopy (SEM). In vitro biological activity was quantified applying L-929 cell line. Ovalbumin (OVA)-challenged balb/c mice were employed to evaluate the anti-TNFα activity of antibody formulation as in vivo experimental model. SD sample consisting of 36 mg trehalose, 12 mg cysteine, and 0.05% of Tween 20 was selected with minimum aggregation/fragmentation rate constants of 0.07 and 0.05 (1/month) based on 1 and 2 months of storage at 40°C and relative humidity of 75%. Fine particle fraction (FPF) value of this formulation was 67.75% with desired particle size and surface morphology for respiratory delivery. EC50 was 8.176 and 6.733 ng/ml for SD Infliximab and Remicade®, respectively. SD antibody reduced TNFα (26.56 pg/ml) secretion in mouse lung tissue, more than 2 orders of magnitudes comparing positive control group (TNFα, 68.34 pg/ml). The success of antibody inhalation mainly depended on the spray drying condition, formulation components, and stability of antibody within aerosolization. Inhaled Infliximab could be a potential drug for local inhibition of lung inflammation.
Assuntos
Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/uso terapêutico , Infliximab/administração & dosagem , Infliximab/uso terapêutico , Administração por Inalação , Animais , Cromatografia em Gel , Estabilidade de Medicamentos , Inaladores de Pó Seco , Excipientes , Luz , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Eletrônica de Varredura , Ovalbumina , Tamanho da Partícula , Espalhamento de Radiação , Espectroscopia de Infravermelho com Transformada de Fourier , Fator de Necrose Tumoral alfa/antagonistas & inibidoresRESUMO
Freeze-dried immunoglobulin G (IgG) incorporating trehalose and human serum albumin (HSA) was statistically evaluated regarding the existence of synergism between additives on the stability profile. The levels of HSA (X1) and trehalose (X2) were independent variables. Aggregation following the process (Y1), after 2 and 3 months at 40°C (Y2) and (Y3), respectively, along with the rate constant of aggregation (Y4) were dependent variables. Aggregation and beta-sheet conformation were quantified through size-exclusion chromatography (SEC-HPLC) and Fourier transform infrared spectroscopy (FTIR). Central composite design (CCD) suggested the best formulation. The integrity and thermodynamic stability of optimized formulation were investigated through sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and differential scanning calorimetry (DSC). The calculated responses were Y1, 0-0.90%; Y2, 0.4-4.3%; Y3, 2.10-13.46%; and Y4, 0.16-0.69 1/month. The optimized formulation had 10 mg IgG, 86 mg trehalose, and 1 mg HSA with observed responses of Y1, 0.01%; Y2, 0.51%; Y3, 3.08%; and Y4, 0.33 1/month. The models were statistically well-fitted. The optimized formulation was amorphous during freeze-drying (FD), and no fragmentation was observed. Trehalose and HSA demonstrated statistical synergism. CCD was successfully employed to recommend the best ratio of stabilizers and achieve the maximum stabilization of IgG as a model freeze-dried antibody.
Assuntos
Desenho de Fármacos , Imunoglobulina G/química , Albumina Sérica Humana/síntese química , Trealose/síntese química , Varredura Diferencial de Calorimetria/métodos , Combinação de Medicamentos , Estabilidade de Medicamentos , Liofilização/métodos , Humanos , Imunoglobulina G/administração & dosagem , Albumina Sérica Humana/administração & dosagem , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Trealose/administração & dosagemRESUMO
OBJECTIVE: To evaluate the effectiveness of topical pentoxifylline (PTX) on pressure ulcer (PU) healing in critically ill patients. METHOD: In this randomised, double blind, placebo-controlled clinical trial, patients with category I or II PUs were randomly assigned to receive either topical PTX 5% or a placebo twice daily for 14 days. Changes in PU characteristics (category and size) were assessed. The category of the PU was determined by the Stirling Pressure Ulcer Severity Scale (two-digit) at baseline (day zero), day seven and day 14 of treatment. PU length and width was measured with a disposable ruler and expressed as cm2. RESULTS: A total of 112 adult patients were enrolled in the study. Median PU size and score at day zero were 32 (10.00-69.33)cm2 and 1(1.00-2.00) respectively. In the PTX group, the mean differences (95% confidence interval, CI) of all PU scores and sizes decreased significantly across the intervals (day seven versus day zero, day 14 versus day zero, and day 14 versus day seven), compared with the placebo group Conclusion: The severity and size of PUs improved significantly in patients who received topical PTX 5% ointment twice a day for 14 days compared with those in the placebo group. Topical PTX may be considered as a potential option in the treatment of categories I and II PUs in critically ill patients.
Assuntos
Pentoxifilina/uso terapêutico , Úlcera por Pressão/tratamento farmacológico , Vasodilatadores/uso terapêutico , Administração Tópica , Método Duplo-Cego , Feminino , Humanos , Unidades de Terapia Intensiva , Masculino , Pessoa de Meia-IdadeRESUMO
OBJECTIVE: The objective of this study was to evaluate the effect of lipid structure on physicochemical properties of chitosan-fatty acid nanomicelles and prepare an optimum ciprofloxacin-loaded formulation from these conjugates which could enhance the antibacterial effects of drug against some important pathogens like P. aeruginosa. SIGNIFICANCE: Nowadays, resistance in infectious diseases is a growing worldwide concern. Nanocarriers can increase the therapeutic index and consequently reduce the antibiotic resistance. By site-specific delivery of drug, the adverse effects of broad-spectrum antibiotics such as ciprofloxacin would be reduced. METHODS: Fatty acid grafted chitosan conjugates were synthetized in the presence of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide. The effects of fatty acid type (stearic acid, palmitic acid, and linoleic acid) on physicochemical properties of conjugates were investigated. Ciprofloxacin was encapsulated in nanomicelles by thin film hydration method. Also, the preparation process was optimized with a central composite design. The antibacterial effect of optimum formulation against P. aeruginosa, K. pneumoniae, and S. pneumoniae species was determined. RESULTS: All conjugates were synthetized with high yield values and the substitution degrees ranged between 2.13 and 35.46%. Ciprofloxacin was successfully encapsulated in nanomicelles. The optimum formulation showed high drug loading (≈ 19%), with particle size of about 260 nm and a sustained release profile of ciprofloxacin. The minimum inhibitory concentrations of ciprofloxacin in optimum formulation against P. aeruginosa and K. pneumoniae species were 4 and 2 times lower in comparison with the free drug, respectively. CONCLUSIONS: The antibacterial effect of ciprofloxacin was improved by encapsulation of drug in chitosan nanomicelles.
Assuntos
Antibacterianos/farmacologia , Quitosana/química , Ciprofloxacina/farmacologia , Portadores de Fármacos/química , Composição de Medicamentos/métodos , Ácidos Graxos/química , Klebsiella pneumoniae/efeitos dos fármacos , Micelas , Testes de Sensibilidade Microbiana , Nanopartículas/química , Pseudomonas aeruginosa/efeitos dos fármacos , Streptococcus pneumoniae/efeitos dos fármacosRESUMO
We aimed to prepare spray-freeze-dried powder of IgG considering physicochemical stability and aerodynamic aspects. Spray-freeze drying (SFD) exposes proteins to various stresses which should be compensated by suitable stabilizers. The competence of cyclodextrins (CDs), namely beta-cyclodextrin (ßCD) and hydroxypropyl ßCD (HPßCD), at very low concentrations, was investigated in the presence of separate mannitol- and trehalose-based formulations. Spray-freeze-dried preparations were quantified in terms of monomer recovery and conformation by size exclusion chromatography (SEC-HPLC) and Fourier transform infrared (FTIR) spectroscopy, respectively. Differential scanning calorimetry (DSC) and X-ray diffractometry (XRD) were employed to identify the thermal characteristics of powders. Particle morphology was visualized by scanning electron microscopy (SEM). Aerodynamic behavior of powders was checked through an Anderson cascade impactor (ACI). Although all formulations protected antibody from aggregation during the SFD process (aggregation < 1%), mannitol-containing ones failed upon the storage (19.54% in the worst case). Trehalose-HPßCD incomparably preserved the formulation with fine particle fraction (FPF) of 51.29%. Crystallization of mannitol resulted in IgG destabilization upon storage. Although employed concentration of CDs is too low (less than 50:1 molar ratio to protein), they successfully served as stabilizing agents in SFD with perfect improvement in aerosol functionality. Graphical Abstract á .
Assuntos
Ciclodextrinas/química , Liofilização/métodos , Imunoglobulina G/química , Trealose/química , Aerossóis , Estabilidade de Medicamentos , Pós/químicaRESUMO
OBJECTIVES: Dry powder formulations are extensively used to improve the stability of antibodies. Spray drying is one of important methods for protein drying. This study investigated the effects of trehalose, hydroxypropyl beta cyclodextrin (HPBCD) and beta cyclodextrin (BCD) on the stability and particle properties of spray-dried IgG. METHODS: D-optimal design was employed for both experimental design and analysis and optimization of the variables. The size and aerodynamic behavior of particles were determined using laser light scattering and glass twin impinger, respectively. In addition, stability, ratio of beta sheets and morphology of antibody were analyzed using size exclusion chromatography, IR spectroscopy and electron microscopy, respectively. RESULTS: Particle properties and antibody stability were significantly improved in the presence of HPBCD. In addition, particle aerodynamic behavior, in terms of fine-particle fraction (FPF), enhanced up to 52.23%. Furthermore, antibody was better preserved not only during spray drying, but also during long-term storage. In contrast, application of BCD resulted in the formation of larger particles. Although trehalose caused inappropriate aerodynamic property, it efficiently decreased antibody aggregation. CONCLUSION: HPBCD is an efficient excipient for the development of inhalable protein formulations. In this regard, optimal particle property and antibody stability was obtained with proper combination of cyclodextrins and simple sugars, such as trehalose.
Assuntos
Anticorpos/química , Ciclodextrinas/química , Pós/química , Trealose/química , beta-Ciclodextrinas/química , Administração por Inalação , Varredura Diferencial de Calorimetria , Química Farmacêutica , Dessecação , Estabilidade de Medicamentos , Excipientes/química , Tamanho da PartículaRESUMO
The influence of poly ethylene glycol (PEG) at different molecular weights (MWs) and ratios was studied on the stability of freeze-dried immune globulin G (IgG). PEGs (600-4000 Dalton) at concentrations of 0.5 and 5% W/V were applied in the presence of 40 and 60% W/W of trehalose to prepare freeze-dried IgG formulations. Size-exclusion chromatography, infra-red spectroscopy, differential scanning calorimeter, and gel electrophoresis were performed to characterize lyophilized samples. Pure IgG demonstrated the highest aggregation of 5.77 ± 0.10% after process and 12.66 ± 0.50% as well as 44.69 ± 0.50% upon 1 and 2 months of storage at 45 °C, respectively. 5% W/V of PEGs 4000 in combination with 40% W/W trehalose, significantly suppressed aggregation, 0.05 ± 0.01%, with minimum aggregation rate constant of 0.32 (1/month). The integrity of IgG molecules and secondary conformation were properly preserved in all formulations comparing native IgG. It could be concluded that appropriate concentration and MW of PEGs, prominently augmented stabilizing effect of trehalose on freeze-dried antibody through inserting additional supportive mechanisms of actions.
Assuntos
Liofilização/métodos , Imunoglobulina G/química , Polietilenoglicóis/química , Trealose/química , Varredura Diferencial de Calorimetria , Química Farmacêutica , Estabilidade de Medicamentos , Imunoglobulina G/metabolismo , Peso MolecularRESUMO
The present study aimed at preparation and optimization of stable freeze-dried immunoglobulin G (IgG) applying proper amount of antibody with efficient combination of trehalose and hydroxypropyl-ß-cyclodextrin (HPßCD). Response surface methodology was employed through a three-factor, three-level Box-Behnken design. Amounts of IgG (X1), trehalose (X2) and HPßCD (X3) were independent variables. Aggregation following process (Y1), after one month at 45 °C (Y2), upon two month at 45 °C (Y3) and beta-sheet content of IgG (Y4) were determined as dependent variables. Results were fitted to quadratic models (except for beta-sheet content), describing the inherent relationship between main factors. Optimized formulation composed of 55.85 mg IgG, 52.51 mg trehalose and 16.01 mg HPßCD was prepared. The calculated responses of the optimized formulation were as follows: Y1 = 0.19%, Y2 = 0.78%, Y3 = 1.88% and Y4 = 68.60%, respectively. The thermal analysis confirmed the amorphous nature of optimum formulation and the integrity of IgG was shown to be favorably preserved. Validation of the optimization study demonstrated high degree of prognostic ability. The DOE study successfully predicted the optimum values of antibody as well as stabilizers for desirable process and storage stabilization of freeze-dried IgG.
Assuntos
Anticorpos , Química Farmacêutica , 2-Hidroxipropil-beta-Ciclodextrina , Emulsões , Tamanho da Partícula , Estabilidade Proteica , TrealoseRESUMO
Pulmonary delivery of peptides remains an important, noninvasive route of administration that is attractive because it offers high bioavailability and patient compliance. Optimization of particle characteristics for deposition in the deep regions of the lung after inhalation and retention of peptide stability are key challenges to their delivery to the lungs. The present study investigated the use of spray freeze-drying to produce porous inhalable parathyroid hormone (1-34)-loaded microparticles suitable for pulmonary delivery. The influence of different excipients in the medium of water or citrate buffer on microparticles characteristics, peptide stability and its systemic delivery in rats were evaluated. Using leucine at 10% (w/w) and hydroxy propyl-ß-cyclodextrin (HPßCD) at 0.04% (w/w) in water or citrate medium preserved parathyroid hormone (1-34) stability by spray freeze-drying. Aerosol performance showed that leucine was more effective than HPßCD in producing inhalable microparticles. Nevertheless, there was no statistical difference between bioavailabilities of HPßCD containing formulations and leucine-containing formulations in the presence of citrate buffer; and even in the presence of water, HPßCD resulted in higher bioavailability compared to leucine. The high absolute bioavailability (up to 47.25%) of formulations could facilitate replacement of injected form of parathyroid hormone (1-34) by dry powder inhaler form.