RESUMEN
RATIONALE: Cystic fibrosis (CF) lung disease is caused by the loss of function of the cystic fibrosis transmembrane conductance regulator (CFTR) combined with hyperactivation of the epithelial sodium channel (ENaC). In the lung, ENaC is responsible for movement of sodium. Hyperactivation of ENaC, which creates an osmotic gradient that pulls fluid out of the airway, contributes to reduced airway hydration, causing mucus dehydration, decreased mucociliary clearance, and recurrent acute bacterial infections. ENaC represents a therapeutic target to treat all patients with CF independent of their underlying CFTR mutation. OBJECTIVES: To investigate the in vitro and in vivo efficacy of SPX-101, a peptide mimetic of the natural regulation of ENaC activity by short palate, lung, and nasal epithelial clone 1, known as SPLUNC1. METHODS: ENaC internalization by SPX-101 in primary human bronchial epithelial cells from healthy and CF donors was assessed by surface biotinylation and subsequent Western blot analysis. SPX-101's in vivo therapeutic effect was assessed by survival of ß-ENaC-transgenic mice, mucus transport in these mice, and mucus transport in a sheep model of CF. MEASUREMENTS AND MAIN RESULTS: SPX-101 binds selectively to ENaC and promotes internalization of the α-, ß-, and γ-subunits. Removing ENaC from the membrane with SPX-101 causes a significant decrease in amiloride-sensitive current. The peptide increases survival of ß-ENaC-transgenic mice to greater than 90% with once-daily dosing by inhalation. SPX-101 increased mucus transport in the ß-ENaC mouse model as well as the sheep model of CF. CONCLUSIONS: These data demonstrate that SPX-101 promotes durable reduction of ENaC membrane concentration, leading to significant improvements in mucus transport.
Asunto(s)
Regulador de Conductancia de Transmembrana de Fibrosis Quística/uso terapéutico , Fibrosis Quística/tratamiento farmacológico , Bloqueadores del Canal de Sodio Epitelial/uso terapéutico , Canales Epiteliales de Sodio/uso terapéutico , Depuración Mucociliar/efectos de los fármacos , Mucosa Respiratoria/efectos de los fármacos , HumanosRESUMEN
The purpose of this research was to prepare a dry powder vaccine formulation containing whole inactivated influenza virus (WIIV) and a mucoadhesive compound suitable for nasal delivery. Powders containing WIIV and either lactose or trehalose were produced by lyophilization. A micro-ball mill was used to reduce the lyophilized cake to sizes suitable for nasal delivery. Chitosan flakes were reduced in size using a cryo-milling technique. Milled powders were sieved between 45 and 125 microm aggregate sizes and characterized for particle size and distribution, morphology, and flow properties. Powders were blended in the micro-ball mill without the ball. Lyophilization followed by milling produced irregularly shaped, polydisperse particles with a median primary particle diameter of approximately 21 microm and a yield of approximately 37% of particles in the 45 to 125 microm particle size range. Flow properties of lactose and trehalose powders after lyophilization followed by milling and sieving were similar. Cryo-milling produced a small yield of particles in the desired size range (<10%). Lyophilization followed by milling and sieving produced particles suitable for nasal delivery with different physicochemical properties as a function of processing conditions and components of the formulation. Further optimization of particle size and morphology is required for these powders to be suitable for clinical evaluation.
Asunto(s)
Vacunas contra la Influenza/administración & dosificación , Administración Intranasal , Liofilización , Tamaño de la Partícula , Polvos , Trehalosa/administración & dosificaciónRESUMEN
The purpose of this research was to prepare a dry powder vaccine formulation containing whole inactivated influenza virus (VIIV) and a mucoadhesive compound suitable for nasal delivery. Powders containing WIIV and either lactose or trehalose were produced by lyophilization. A micro-ball mill was used to reduce the lyophilized cake to sizes suitable for nasal delivery. Chitosan flakes were reduced in size using a cryo-milling technique. Milled powders were sieved between 45 and 125 µm aggregate sizes and characterized for particle size and distribution, morphology, and flow properties. Powders were blended in the micro-ball mill without the ball. Lyophilization followed by milling produced irregularly shaped, polydisperse particles with a median primary particle diameter of ≈21 µm and a yield of ≈37% of particles in the 45 to 125 µm particle size range. Flow properties of lactose and trehalose powders after lyophilization followed by milling and sieving were similar. Cryo-milling produced a small yield of particles in the desired size range (<10%). Lyophilization followed by milling and sieving produced particles suitable for nasal delivery with different physicochemical properties as a function of processing conditions and components of the formulation. Further optimization of particle size and morphology is required for these powders to be suitable for clinical evaluation.
RESUMEN
Dry powder inhalers (DPIs) have been in use since the 1970s, but it is only within the past few years that their use has constituted > approximately 10% of the inhaler units sold worldwide. Similarly, active DPIs have been in development for more than a decade, but no active device has yet been approved. Oriel is developing an active DPI technology that uses a very simple physical design coupled with a complex knowledge of powder flow and dispersion characterisation. The DPI uses electromechanical vibration with frequencies determined through the analysis of powder flow properties. Results so far have shown highly reproducible, efficient performance. The technology lends itself to both unit-dose and multidose platforms in a targeted cost-effective DPI.
Asunto(s)
Sistemas de Liberación de Medicamentos/métodos , Enfermedades Pulmonares/tratamiento farmacológico , Nebulizadores y Vaporizadores , Tecnología Farmacéutica , Aerosoles , Sistemas de Liberación de Medicamentos/economía , Sistemas de Liberación de Medicamentos/instrumentación , Electricidad , Diseño de Equipo , Humanos , Nebulizadores y Vaporizadores/economía , Polvos , VibraciónRESUMEN
Recent findings indicate that the inhalation of large manufactured porous particles may be particularly effective for drug delivery. In this study, a mathematical model was employed to systematically investigate the effects of particle size, particle density, aerosol polydispersity, and patient ventilatory parameters on deposition patterns of inhaled drugs in healthy human lungs. Aerodynamically similar particles with densities of 0.1, 1.0, and 2.0 g/cm(3) were considered. Particle size distributions were defined with mass median aerodynamic diameters (MMADs) ranging from 1 to 3 microm and geometric standard deviations ranging from 1.5 to 2.5, representing particles in the respirable size range. Breathing rates of 30 and 60 L/min with tidal volumes of 500 to 3000 mL were assumed, simulating shallow to deep breaths from a dry powder inhaler. Particles with a high density and a small geometric diameter had slightly greater deposition fractions than particles that were aerodynamically similar, but had lower density and larger geometric size (typical of manufactured porous particles). This can be explained by the fact that particles with a small geometric diameter deposit primarily by diffusion, which is a function of geometric size but is independent of density. As MMAD increased, the effect of density on deposition was less pronounced because of the decreased efficiency of diffusion for large particles. These data suggest that polydisperse aerosols containing a significant proportion of submicron particles will deposit in the pulmonary airways with greater efficiency than aerodynamically similar aerosols comprised of geometrically larger porous particles.
Asunto(s)
Aerosoles/química , Modelos Biológicos , Administración por Inhalación , Aerosoles/administración & dosificación , Aerosoles/farmacocinética , Simulación por Computador , Sistemas de Liberación de Medicamentos/métodos , Humanos , Pulmón/metabolismo , Pulmón/fisiología , Modelos Químicos , Porosidad , Volumen de Ventilación Pulmonar/fisiologíaRESUMEN
A novel light emitting diode (LED) array-based light induced fluorescence (LIF) sensor is presented as an analytical methodology for at-line cleaning verification within the pharmaceutical industry. This sensor differs from conventional LIF sensors through the ability to dynamically control both the LED excitation array and detection parameters, enabling the exploitation of the optical power and detection sensitivity to rapidly detect trace concentrations of residual drug. This feature makes this sensor an ideal alternative to conventional cleaning verification analytical methodologies. In this study, the LIF sensor was validated as an analytical technique through the analysis of specificity, precision, linearity, limit of quantitation, and accuracy, with respect to solutions and swab extracts of a single pharmaceutical compound (Compound A). The validated system was then utilized for cleaning process optimization and subsequent routine cleaning process verification following three manufacturing campaigns. The LIF sensor enabled a significant improvement in the analysis time for quantitative detection of Compound A; individual swab and rinsate extracts were analyzed in less than 1 min. The results presented herein effectively demonstrate the ability of the novel LIF sensor to efficiently function as a valid at-line analytical methodology for cleaning verification.
Asunto(s)
Industria Farmacéutica/métodos , Fluorescencia , Luz , Tecnología Farmacéutica/métodos , Sensibilidad y EspecificidadRESUMEN
Key factors that contribute to the aerodynamic properties of aerosol particles are found in Stokes' law. These factors may be monitored or controlled to optimize drug delivery to the lungs. Predictions of the aerodynamic behavior of therapeutic aerosols can be derived in terms of the physical implications of particle slip, shape and density. The manner in which each of these properties have been used or studied by pharmaceutical scientists to improve lung delivery of drugs is readily understood in the context of aerosol physics. Additional improvement upon current aerosol delivery of particulates may be predicted by further theoretical scrutiny.
Asunto(s)
Sistemas de Liberación de Medicamentos/métodos , Pulmón/efectos de los fármacos , Polvos/administración & dosificación , Administración por Inhalación , Animales , Química Farmacéutica , Humanos , Nebulizadores y Vaporizadores , Tamaño de la Partícula , Polvos/química , Tecnología Farmacéutica/métodosRESUMEN
Intranasal (i.n.) vaccination represents an attractive non-invasive alternative to needle-based injection and provides superior protection at mucosal surfaces. However, new formulations are needed to improve efficacy and reduce the refrigerated storage and distribution requirements associated with standard liquid vaccines. Here, we describe a powder formulation of whole inactivated influenza virus and a novel i.n. delivery platform. The powder-formulated vaccine elicited a significant serum antibody response in rats that was at least as strong as that provided by the liquid vaccine administered i.n. or via intramuscular (i.m.) injection. Significant nasal IgA responses were also observed solely after i.n. delivery. This study demonstrates for the first time the generation of potent nasal mucosal and systemic immune responses using an i.n. delivered influenza vaccine powder and suggests an alternative approach to vaccination against influenza and other infectious diseases.