Development and characterization of a spray-dried inhalable ternary combination for the treatment of Pseudomonas aeruginosa biofilm infection in cystic fibrosis.

Cystic fibrosis (CF) is an inherited lung disease characterised by the accumulation of thick layers of dried mucus in the lungs which serve as a nidus for chronic infection. Pseudomonas aeruginosa is the predominant cause of chronic lung infection in cystic fibrosis. The dense mucus coupled with biofilm formation hinder antibiotic penetration and prevent them from reaching their target. Mucoactive agents are recommended in the treatment of CF in combination with antibiotics. In spite of the extensive research in developing novel drug combinations for the treatment of lung infection in CF, to our knowledge, there is no study that combines antibiotic, antibiofilm and mucoactive agent in a single inhaled dry powder formulation. In the present study, we investigate the possibility of adding a mucoactive agent to our previously developed ciprofloxacinquercetin (antibiotic-antibiofilm) dry powder for inhalation. Three mucoactive agents, namely mannitol (MAN), N-acetyl-L-cysteine (NAC) and ambroxol hydrochloride (AMB), were investigated for this purpose. The ternary combinations were prepared via spray drying without the addition of excipients. All ternary combinations conserved or improved the antibacterial and biofilm inhibition activities of ciprofloxacin against P. aeruginosa (ATCC 10145). The addition of AMB resulted in an amorphous ternary combination (SD-CQA) with superior physical stability as indicated by DSC and nonambient XRPD. Furthermore, SD-CQA displayed better in vitro aerosolization performance (ED ∼ 71 %; FPF ∼ 49 %) compared to formulations containing MAN and NAC (ED ∼ 64 % and 44 %; FPF ∼ 44 % and 29 %, respectively). In conclusion, a ternary drug combination powder with suitable aerosolization, physical stability and antibacterial/antibiofilm properties was prepared by a single spray drying step.

Quality by design - Spray drying of ciprofloxacin-quercetin fixed-dose combination intended for inhalation.

Spray drying is a well-suited technique for producing fixed-dose drug combinations. There has been a growing interest in utilizing spray drying to engineer carrier-free inhalable drug particles. The aim of this study was to understand and optimise the spray drying process of a ciprofloxacin-quercetin fixed dose combination intended for pulmonary administration. A 24-1 fractional factorial design and multivariate data analysis was used to identify important process parameters and investigate correlations with particle characteristics. The independent variables were solute concentration along with the processing parameters: solution flow rate, atomizing air flow rate and inlet temperature. The dependent variables included particle size distribution, yield and residual moisture content (RMC). Correlations between dependent and independent variables were further investigated via principal component analysis. Overall, solution flow rate, atomizing air flow rate and inlet temperature were found to affect the particle size D(v,50) and D(v,90) while the solute concentration and the atomizing air flow rate mainly affected the span. The inlet temperature was the most important parameter affecting the RMC and the yield. The formulation with optimized independent variables had a D(v,50) and span values of 2.42 µm and 1.81 with excellent process yield >70% and low RMC i.e. 3.4%. The optimized formulation was further investigated for its in vitro aerosolization performance using next generation impactor (NGI); it exhibited high emitted dose (ED > 80%) and fine particle fractions (FPF > 70%) for both drugs.

Development and characterization of a spray-dried inhalable ciprofloxacin-quercetin co-amorphous system.

Spray drying is an increasingly used particle engineering technique for the production of dry powders for inhalation. However, the amorphous nature of most spray-dried particles remains a big challenge affecting both the chemical and the physical stability of the dried particles. Here, we study the possibility of producing co-amorphous ciprofloxacin-quercetin inhalable particles with improved amorphous stability compared to the individual amorphous drugs. Ciprofloxacin (CIP), a broad-spectrum antibiotic, was co-spray dried with quercetin (QUE), a compound with antibiofilm properties, from an ethanol-water co-solvent system at 2:1, 1:1 and 1:2 M ratios to investigate the formation of co-amorphous CIP-QUE particles. Differential scanning colorimetry (DSC) and X-ray powder diffraction (XRPD) were used for solid-state characterization; dynamic vapor sorption (DVS) was used for investigating the moisture sorption behaviour. The intermolecular interaction was studied via solution-state nuclear magnetic resonance (NMR) and Fourier transform infrared (FTIR) spectroscopy; the miscibility of the drugs was predicted via free energy calculations based on the Flory-Huggins interaction parameter (χ). A next generation impactor (NGI) was used to study the in vitro aerosol performance of the spray-dried powders. The physicochemical characteristics such as particle size, density, morphology, cohesion, water content and saturation solubility of the spray-dried powders were also studied. The co-spray-dried CIP-QUE powders prepared at the three molar ratios were predominantly amorphous. However, differences were observed between sample types. It was found that at a molar ratio of 1:1, CIP and QUE form a single co-amorphous system. However, increasing the molar ratio of either drug results in the formation of an additional amorphous phase, formed from the excess of the corresponding drug. Despite these differences, DVS showed that elevated humidity had a much lower influence on all three co-amorphous systems compared with the individual amorphous drugs. In vitro aerosolization study showed co-deposition of the two drugs from CIP-QUE powders with a desirable aerosol performance (ED âˆ¼ 72-94%; FPF âˆ¼ 48-65%) whereas QUE-only amorphous powder had an ED of 36% and a FPF of 22%. In summary, spray-dried CIP-QUE combinations resulted in co-amorphous systems with boosted stability and improved aerosol performance with the 1:1 M ratio exhibiting the greatest improvement.

Developing ciprofloxacin dry powder for inhalation: A story of challenges and rational design in the treatment of cystic fibrosis lung infection.

Cystic fibrosis (CF) is an inherited multisystem disease affecting the lung which leads to a progressive decline in lung function as a result of malfunctioning mucociliary clearance and subsequent chronic bacterial infections. Pseudomonas aeruginosa is the predominant cause of lung infection in CF patients and is associated with significant morbidity and mortality. Thus, antibiotic therapy remains the cornerstone of the treatment of CF. Pulmonary delivery of antibiotics for lung infections significantly reduces the required dose and the associated systemic side effects while improving therapeutic outcomes. Ciprofloxacin is one of the most widely used antibiotics against P. aeruginosa and the most effective fluoroquinolone. However, in spite of the substantial amount of research aimed at developing ciprofloxacin powder for inhalation, none of these formulations has been commercialized. Here, we present an integrated view of the diverse challenges associated with delivering ciprofloxacin dry particles to the lungs of CF patients and the rationales behind recent formulations of ciprofloxacin dry powder for inhalation. This review will discuss the challenges in developing ciprofloxacin powder for inhalation along with the physiological and pathophysiological challenges such as ciprofloxacin lung permeability, overproduction of viscous mucus and bacterial biofilms. The review will also discuss the current and emerging particle engineering approaches to overcoming these challenges. By doing so, we believe the review will help the reader to understand the current limitations in developing an inhalable ciprofloxacin powder and explore new opportunities of rational design strategies.

Leucine as an excipient in spray dried powder for inhalation.

Leucine is a promising excipient with several applications in the development of inhalable spray-dried powder of high- and low-dose drugs. The addition of leucine has exhibited significant enhancing effects on the aerosolization and physical stability of the produced particles. Here, we focus not only on the applications of leucine in inhalable spray-drying powders, but also on the underlying mechanisms by which the formulation and processing parameters dictate the behavior of leucine during the drying process and, therefore, its functionalities within the dried powder. Additionally, we highlight the current regulatory status of leucine. Such insights are important for more efficient utilization of leucine in the future, both for dry powder inhaler formulations and other pharmaceutical applications.

Investigation into the Solid-State Properties and Dissolution Profile of Spray-Dried Ternary Amorphous Solid Dispersions: A Rational Step toward the Design and Development of a Multicomponent Amorphous System.

The formulation of oral amorphous solid dispersions (ASD) includes the use of excipients to improve physical stability and enhance bioavailability. Combinations of excipients (polymers and surfactants) are often employed in pharmaceutical products to improve the delivery of poorly water-soluble drugs. However, additive interactions in multicomponent ASD systems have not been extensively studied and may promote crystallization in an unpredictable manner, which in turn may affect the physical stability and dissolution profile of the product. The main aim of this study was to understand the effect of different surfactant and polymer combinations on the solid-state properties and dissolution behavior of ternary spray-dried solid dispersions of dipyridamole and cinnarizine. The surfactants chosen for this study were sodium dodecyl sulfate and poloxamer 188, and the model polymers used were polyvinylpyrrolidone K30 and hydroxypropyl methylcellulose K100. The spray-dried ternary dispersions maintained higher supersaturation compared to either the crystalline drug equilibrium solubility or their respective physical mixtures. However, rapid and variable dissolution behavior was observed for different formulations. The maximum supersaturation level was observed with drug-polymer-polymer ternary dispersions. On the other hand, incorporating the surfactant into binary (drug-polymer) and ternary (drug-polymer-polymer) ASDs adversely affected the physical stability and dissolution by promoting crystallization. On the basis of these observations, a thorough investigation into the impact of combinations of additives on amorphous drug crystallization during dissolution and stability studies is recommended in order to develop optimized formulations of supersaturating dosage forms.

Understanding the generation and maintenance of supersaturation during the dissolution of amorphous solid dispersions using modulated DSC and 1H NMR.

In this study, the dissolution behaviour of dipyridamole (DPM) and cinnarizine (CNZ) spray-dried amorphous solid dispersions (ASDs) using polyvinyl pyrrolidone (PVP) and polyacrylic acid (PAA) as a carrier matrix were evaluated and compared. The drug concentrations achieved from the dissolution of PVP and PAA solid dispersions were significantly greater than the equilibrium solubility of crystalline DPM and CNZ in phosphate buffer pH 6.8 (PBS 6.8). The maximum drug concentration achieved by dissolution of PVP and PAA solid dispersions did not exceed the theoretically calculated apparent solubility of amorphous DPM and CNZ. However, the degree of supersaturation of DPM and CNZ increased considerably as the polymer weight fraction within the solid dispersion increased. In addition, the supersaturation profile of DPM and CNZ were studied in the presence and absence of the polymers. PAA was found to maintain a higher level of supersaturation compared to PVP. The enhanced drug solution concentration following dissolution of ASDs can be attributed to the reduced crystal growth rates of DPM and CNZ at an equivalent supersaturation. We have also shown that, for drugs having high crystallization tendency and weak drug-polymer interaction, the feasible way to increase dissolution might be increase the polymer weight fraction in the ASD. Solution 1H NMR spectra were used to understand dissolution mechanism and to identify drug-polymer interaction. The change in electron densities of proton attached to different groups in DPM and CNZ suggested drug-polymer interaction in solution. The relative intensities of peak shift and nature of interaction between drug and polymer in different systems are different. These different effects suggest that DPM and CNZ interacts in a different way with PVP and PAA in solution which goes some way towards explaining the different polymeric effect, particularly in terms of inhibition of drug recrystallization and dissolution of DPM and CNZ ASDs. These results established that the different drug/polymer interactions in the solid state and in solution give rise to the variation in dissolution profile observed for different systems.

Theoretical and experimental investigation of drug-polymer interaction and miscibility and its impact on drug supersaturation in aqueous medium.

Amorphous solid dispersions (ASDs) have the potential to offer higher apparent solubility and bioavailability of BCS class II drugs. Knowledge of the solid state drug-polymer solubility/miscibility and their mutual interaction are fundamental requirements for the effective design and development of such systems. To this end, we have carried out a comprehensive investigation of various ASD systems of dipyridamole and cinnarizine in polyvinylpyrrolidone (PVP) and polyacrylic acid (PAA) at different drug loadings. Theoretical and experimental examinations (by implementing binary and ternary Flory-Huggins (F-H) theory) related to drug-polymer interaction/miscibility including solubility parameter approach, melting point depression method, phase diagram, drug-polymer interaction in the presence of moisture and the effect of drug loading on interaction parameter were performed. The information obtained from this study was used to predict the stability of ASDs at different drug loadings and under different thermal and moisture conditions. Thermal and moisture sorption analysis not only provided the composition-dependent interaction parameter but also predicted the composition dependent miscibility. DPM-PVP, DPM-PAA and CNZ-PAA systems have shown molecular level mixing over the complete range of drug loading. For CNZ-PVP, the presence of a single Tg at lower drug loadings (10, 20 and 35%w/w) indicates the formation of solid solution. However, drug recrystallization was observed for samples with higher drug weight fractions (50 and 65%w/w). Finally, the role of polymer in maintaining drug supersaturation has also been explored. It has been found that drug-polymer combinations capable of hydrogen-bonding in the solution state (DPM-PVP, DPM-PAA and CNZ-PAA) are more effective in preventing drug crystallization compared to the drug-polymer systems without such interaction (CNZ-PVP). The DPM-PAA system outperformed all other ASDs in various stability conditions (dry-state, in the presence of moisture and in solution state), which was attributed to the drug's low crystallization tendency, the strong DPM-PAA interaction, the robustness of this interaction against moisture or water and the ability of PAA in maintaining DPM supersaturation.

An investigation into the crystallization tendency/kinetics of amorphous active pharmaceutical ingredients: A case study with dipyridamole and cinnarizine.

Amorphous drug formulations have great potential to enhance solubility and thus bioavailability of BCS class II drugs. However, the higher free energy and molecular mobility of the amorphous form drive them towards the crystalline state which makes them unstable. Accurate determination of the crystallization tendency/kinetics is the key to the successful design and development of such systems. In this study, dipyridamole (DPM) and cinnarizine (CNZ) have been selected as model compounds. Thermodynamic fragility (mT) was measured from the heat capacity change at the glass transition temperature (Tg) whereas dynamic fragility (mD) was evaluated using methods based on extrapolation of configurational entropy to zero [Formula: see text] , and heating rate dependence of Tg [Formula: see text] . The mean relaxation time of amorphous drugs was calculated from the Vogel-Tammann-Fulcher (VTF) equation. Furthermore, the correlation between fragility and glass forming ability (GFA) of the model drugs has been established and the relevance of these parameters to crystallization of amorphous drugs is also assessed. Moreover, the crystallization kinetics of model drugs under isothermal conditions has been studied using Johnson-Mehl-Avrami (JMA) approach to determine the Avrami constant 'n' which provides an insight into the mechanism of crystallization. To further probe into the crystallization mechanism, the non-isothermal crystallization kinetics of model systems were also analysed by statistically fitting the crystallization data to 15 different kinetic models and the relevance of model-free kinetic approach has been established. The crystallization mechanism for DPM and CNZ at each extent of transformation has been predicted. The calculated fragility, glass forming ability (GFA) and crystallization kinetics are found to be in good correlation with the stability prediction of amorphous solid dispersions. Thus, this research work involves a multidisciplinary approach to establish fragility, GFA and crystallization kinetics as stability predictors for amorphous drug formulations.

Polymeric Amorphous Solid Dispersions: A Review of Amorphization, Crystallization, Stabilization, Solid-State Characterization, and Aqueous Solubilization of Biopharmaceutical Classification System Class II Drugs.

Poor water solubility of many drugs has emerged as one of the major challenges in the pharmaceutical world. Polymer-based amorphous solid dispersions have been considered as the major advancement in overcoming limited aqueous solubility and oral absorption issues. The principle drawback of this approach is that they can lack necessary stability and revert to the crystalline form on storage. Significant upfront development is, therefore, required to generate stable amorphous formulations. A thorough understanding of the processes occurring at a molecular level is imperative for the rational design of amorphous solid dispersion products. This review attempts to address the critical molecular and thermodynamic aspects governing the physicochemical properties of such systems. A brief introduction to Biopharmaceutical Classification System, solid dispersions, glass transition, and solubility advantage of amorphous drugs is provided. The objective of this review is to weigh the current understanding of solid dispersion chemistry and to critically review the theoretical, technical, and molecular aspects of solid dispersions (amorphization and crystallization) and potential advantage of polymers (stabilization and solubilization) as inert, hydrophilic, pharmaceutical carrier matrices. In addition, different preformulation tools for the rational selection of polymers, state-of-the-art techniques for preparation and characterization of polymeric amorphous solid dispersions, and drug supersaturation in gastric media are also discussed.

Characterisation of dry powder inhaler formulations using atomic force microscopy.

Inhalation formulations are a popular way of treating the symptoms of respiratory diseases. The active pharmaceutical ingredient (API) is delivered directly to the site of action within the deep lung using an inhalation device such as the dry powder inhaler (DPI). The performance of the formulation and the efficiency of the treatment depend on a number of factors including the forces acting between the components. In DPI formulations these forces are dominated by interparticulate interactions. Research has shown that adhesive and cohesive forces depend on a number of particulate properties such as size, surface roughness, crystallinity, surface energetics and combinations of these. With traditional methods the impact of particulate properties on interparticulate forces could be evaluated by examining the bulk properties. Atomic force microscopy (AFM), however, enables the determination of local surface characteristics and the direct measurement of interparticulate forces using the colloidal probe technique. AFM is considered extremely useful for evaluating the surface topography of a substrate (an API or carrier particle) and even allows the identification of crystal faces, defects and polymorphs from high-resolution images. Additionally, information is given about local mechanical properties of the particles and changes in surface composition and energetics. The assessment of attractive forces between two bodies is possible by using colloidal probe AFM. This review article summarises the application of AFM in DPI formulations while specifically focussing on the colloidal probe technique and the evaluation of interparticulate forces.

Ordered DNA wrapping switches on luminescence in single-walled nanotube dispersions.

An extensive study of the time dependence of DNA wrapping in single-walled nanotube (SWNT) dispersions has been carried out, revealing a number of unusual phenomena. SWNTs were dispersed in water with salmon testes DNA and monitored over a three-month period. Between 20 and 50 days after the sample was first prepared, the SWNT photoluminescence (PL) intensity was observed to increase by a factor of 50. This increase was accompanied by a considerable sharpening of the van Hove absorption peaks. High-resolution transmission electron microscopy (HRTEM) images showed the progressive formation of a coating of DNA on the walls of the nanotubes over the three-month period. HRTEM and circular dichroism spectroscopy studies showed that the improvement in both the NIR PL intensity and the van Hove absorption peaks coincided with the completion of a monolayer coating of DNA on the SWNT walls. HRTEM images clearly showed the DNA wrapping helically around the SWNTs in a surprisingly ordered fashion. We suggest that the initial quenching of NIR photoluminescence and broadening of absorption peaks is related to the presence of protonated surface oxides on the nanotubes. The presence of an ordered DNA coating on the nanotube walls mediates both deprotonation and removal of the surface oxides. An extensive DNA coating is required to substantially restore the photoluminescence, and thus, the luminescence switch-on and subsequent saturation indicate the completion of the DNA-wrapping process. The temperature dependence of the PL switch-on, and thus of the wrapping process, was investigated by measuring as functions of temperature both the time before PL switch-on and the time required for the PL intensity to saturate. This allowed the calculation of the activation energies for both the process preceding PL switch-on and the process limiting the rise of PL intensity, which were found to be 31 and 41 kJ mol (-1), respectively. The associated entropies of activation were -263 and -225 J mol (-1) K (-1), respectively. These negative activation entropies suggest that the rate-limiting step is characterized by a change in the system from a less-ordered to a more-ordered state, consistent with the formation of an ordered DNA coating.

Spontaneous exfoliation of single-walled carbon nanotubes dispersed using a designed amphiphilic peptide.

We have observed concentration dependent exfoliation of single-walled carbon nanotubes dispersed in solutions of the synthetic peptide nano-1. As the nanotube concentration is reduced, the bundle diameters tend to decrease before saturating at <2.0 nm for concentrations below 6 x 10(-3) mg/mL. The fraction of individual nanotubes increases with decreasing concentration, saturating at approximately 95% at low concentration. This concentration dependent exfoliation happens even if the dispersions are not sonicated on dilution, albeit over a longer time scale. The populations both of individual nanotubes and of bundles are much higher than expected at high concentrations, indicating the presence of repulsive internanotube interactions stabilizing the dispersions.