Novel Nano-Composite Particles For Controlled Release Drugs Via Inhalation: Liposomal Nanocrystalline Ciprofloxacin

This thesis evaluates the feasibility of developing inhalable dry powders of ciprofloxacin nanocrystals inside liposomes (CNL) by spray drying for controlled drug release. Spray drying was optimized for the production of CNL powder using an experimental design methodology followed by testing the storage stability of the optimized formulations. Dry powders of CNL were successfully produced either by freeze-thaw liposomal liquid formulations followed by spray drying or by a simple one-step process of spray drying. Regardless of the production pathway, sucrose to lipids at 2:1 w/w ratio was an excellent lyoprotectant for liposomes during processing. Drug encapsulation efficiency (EE) was above 79% with mostly a single drug nanocrystal residing inside each liposomal vesicle. The powders composed of spherical particles with a median diameter of ~ 1 µm and a respirable fraction (% wt. < 5 µm) above 58%. A prolonged drug release over 12 hours was achieved in vitro by CNL powders. The optimization of spray drying was performed using the Box–Behnken design. The study independent variables were the protectant type (sucrose, trehalose or lactose), protectant amount, drying temperature, and spray gas flow. Liposomal particle size, drug EE, liposomal zeta potential, and nanocrystal dimensions were the dependant variables. Sucrose and lactose were superior protectants to liposomes during spray drying compared to trehalose. The protectant amount significantly affected the characteristics of CNL powders. The optimized powders were stored for six months at room temperature (RT) and either 4 or 20% relative humidity (RH) for the stability study. Sucrose CNL powder exhibited consistent liposomal integrity, aerosol performance, and controlled release of ciprofloxacin over six months of storage at 4 %RH. However, storage of the powder at 20 %RH for the same period caused sucrose crystallization and consequently a significant change in the physicochemical properties. Lactose CNL maintained superior aerosol performance irrespective of the storage RH. However, liposomal instability occurred at both RHs within the first month of storage. In conclusion, the CNL powders were suitable for aerosol delivery and amenable to a once-daily treatment regimen. The optimum processing conditions utilized 57% w/w sucrose, 80°C inlet temperature, and atomization rate of 742 L/hr. This research suggests that CNL powder formulation with sucrose as a stabilizing excipient must be stored under a relatively low humidity environment of 4 %RH and controlled RT storage to achieve at least a six-month shelf-life

Technegas, A Universal Technique for Lung Imaging in Nuclear Medicine: Technology, Physicochemical Properties, and Clinical Applications

Technegas was developed in Australia as an imaging radioaerosol in the late 1980s and is now commercialized by Cyclomedica, Pty Ltd. for diagnosing pulmonary embolism (PE). Technegas is produced by heating technetium-99m in a carbon crucible for a few seconds at high temperatures (2750 °C) to generate technetium–carbon nanoparticles with a gas-like behaviour. The submicron particulates formed allow easy diffusion to the lung periphery when inhaled. Technegas has been used for diagnosis in over 4.4 m patients across 60 countries and now offers exciting opportunities in areas outside of PE, including asthma and chronic obstructive pulmonary disease (COPD). The Technegas generation process and the physicochemical attributes of the aerosol have been studied over the past 30 years in parallel with the advancement in different analytical methodologies. Thus, it is now well established that the Technegas aerosol has a radioactivity aerodynamic diameter of <500 nm and is composed of agglomerated nanoparticles. With a plethora of literature studying different aspects of Technegas, this review focuses on a historical evaluation of the different methodologies’ findings over the years that provides insight into a scientific consensus of this technology. Also, we briefly discuss recent clinical innovations using Technegas and a brief history of Technegas patents

Storage stability of inhalable, controlled-release powder formulations of ciprofloxacin nanocrystal-containing liposomes

Novel inhalable and controlled release powder formulations of ciprofloxacin nanocrystals inside liposomes (CNL) were recently developed. In the present study, the storage stability of CNL powders consisting of lyoprotectant (i.e. sucrose or lactose), lipids, ciprofloxacin ( CIP ), and magnesium stearate or isoleucine was investigated. These powders were produced by spray drying, collected in a dry box at < 15% relative humidity (RH), then stored at room temperature and either 4 or 20 %RH. Liposomal integrity, CIP encapsulation efficiency (EE), in vitro drug release (IVR), aerosol performance, and solid-state properties were examined over six months. Sucrose CNL powder exhibited consistent liposomal integrity, aerosol performance, and controlled release of CIP over six months of storage at 4 %RH. However, storage of the powder at 20 %RH for the same period caused sucrose crystallization and consequently a significant drop in EE and aerosol performance (p-values <0.05), along with the IVR of CIP becoming similar to that of the non-crystalline CIP liposomal dispersions (f2 > 50). Lactose CNL maintained superior aerosol performance over the six months irrespective of the storage RH. However, liposomal instability occurred at both RHs within the first month of storage with a significant drop in EE and an increase in liposome size (p-values <0.05). Moreover, the IVR assay of CIP from lactose CNL showed a less controlled release and a substantial difference (f2 < 50) from its initial value after six months regardless of the storage RHs. In conclusion, dry powder inhalers of CNL were physiochemically stable in sucrose lyoprotectant when stored below 4 %RH at room temperature for six months

Modeling of a spray drying method to produce ciprofloxacin nanocrystals inside the liposomes utilizing a response surface methodology: Box-Behnken experimental design

Spray drying was previously used to modify the physical form of the encapsulated ciprofloxacin drug to produce ciprofloxacin nanocrystals inside the liposomes (CNL). The purpose of the present study was to optimize CNL powder production by evaluating the response surface via design of experiments (DoE). Using the Box–Behnken (BB) design, the study independent variables were the protectant type (sucrose, trehalose or lactose), protectant amount, drying temperature, and spray gas flow. Individual spray drying experiments were performed at various set points for each variable followed by characterization of the produced powders. Liposomal particle size, drug encapsulation efficiency (EE%), liposomal surface zeta potential, and nanocrystal dimensions were the design dependant variables. By applying the least square regression method on the experimental data, mathematical models were developed using the mathematical software package MATLAB R2018b. Model reliability and the significance of the model’s factors were estimated using analysis of variance (ANOVA). The generated CNL powders showed spherical to elliptical liposomal vesicles with particle sizes ranging from 98 to 159 nm. The EE (%) ranged from 30 to 95% w/w while the zeta potential varied between -3.5 and -10.5 mV. The encapsulated ciprofloxacin nanocrystals were elongated cylindrical structures with an aspect ratio of 4.0 - 7.8. Coefficients of determination ( R 2 > 0.9) revealed a good agreement between the predicted and experimental values for all responses except for the nanocrystal dimensions. Sucrose and lactose were superior to trehalose in protecting the liposomes during spray drying. The amount of sugar significantly affected the characteristics of the CNL powders ( p -value < 0.05). In conclusion, the DoE approach using BB design has efficiently modelled the generation of CNL by spray drying. The optimum processing conditions which produced high drug encapsulation (90%) after formation of nanocrystals and a vesicle size of ~125 nm utilized 57% (w/w) sucrose, an 80°C inlet temperature, and an atomization rate of 742 L/hr

Formation of ciprofloxacin nanocrystals within liposomes by spray drying for controlled release via inhalation

The present study was conducted to harness spray drying technology as a novel method of producing Ciprofloxacin nanocrystals inside liposomes (CNL) for inhalation delivery. Liposomal ciprofloxacin dispersions were spray dried with sucrose as a lyoprotectant in different mass ratios (0.5:1, 1:1 and 2:1 sucrose to lipids), along with 2% w/w magnesium stearate and 5% w/w isoleucine as aerosolization enhancers. Spray drying conditions were: inlet air temperature 50 °C, outlet air temperature 33-35 °C, atomizer rate 742 L/h and aspirator 35 m3/h. After spray drying, the formation of ciprofloxacin nanocrystals inside the liposomes was confirmed by cryo- transmission electron microscopy. The physiochemical characteristics of the spray dried powder (particle size, morphology, crystallinity, moisture content, drug encapsulation efficiency (EE), in vitro aerosolization performance and drug release) were determined. The EE of the liposomes was found to vary between 44 and 87% w/w as the sucrose content was increased from 25 to 57% w/w. The powders contained partially crystalline particles with a volume median diameter of ~ 1 μm. The powders had low water content (~2 % wt.) and were stable at high relative humidity. Aerosol delivery using the Osmohaler® inhaler at a flow rate of 100 L/min produced an aerosol fine particle fraction (% wt. < 5 μm) of 58 - 64%. The formulation with the highest sucrose content (2:1 w/w sucrose to lipid) demonstrated extended ciprofloxacin release from liposomes (80% released within 7 hours) in comparison to the original liquid formulation (80% released within 2 hours). In conclusion, a stable and inhalable CNL powder with controlled drug release was successfully prepared by spray drying

Ciprofloxacin nanocrystals liposomal powders for controlled drug release via inhalation

This study was conducted to evaluate the feasibility of developing inhalable dry powders of liposomal encapsulated ciprofloxacin nanocrystals (LECN) for controlled drug release. Dry powders of LECN were produced by freeze-thaw followed by spray drying. The formulations contained sucrose as a lyoprotectant in different weight ratios (0.75:1, 1:1 and 2:1 sucrose to lipids), along with 2 % magnesium stearate and 5 % isoleucine as aerosolization enhancers. The powder physical properties (particle size, morphology, crystallinity, moisture content), in vitro aerosolization performance, drug encapsulation efficiency and in vitro drug release were investigated. The spray dried powders were comprised of spherical particles with a median diameter of ~ 1 μm, partially crystalline, with a low water content (~2 % mass) and did not undergo recrystallization at high relative humidity. When dispersed by an Osmohaler® inhaler at 100 L/min, the powders showed a high aerosol performance with a fine particle fraction (% wt. < 5 μm) of 66-70 %. After reconstitution of the powders in saline, ciprofloxacin nanocrystals were confirmed by cryoelectron microscopy. The drug encapsulation efficiency of the reconstituted liposomes was 71-79 % compared with the stock liquid formulation. Of the three formulations, the one containing a sucrose to lipids wt. ratio of 2:1 demonstrated a prolonged release of ciprofloxacin from the liposomes. In conclusion, ciprofloxacin nanocrystal liposomal powders were prepared that were suitable for inhalation aerosol delivery and controlled drug release

Nanoscale probing of liposome encapsulating drug nanocrystal using Atomic Force Microscopy-Infrared Spectroscopy

Use of liposomes encapsulating drug nanocrystals for the treatment of diseases like cancer and pulmonary infections is gaining attention. The potential therapeutic benefit of these engineered formulations relies on maintaining the physical integrity of the liposomes and the stability of the encapsulated drug. With the significant advancement in the microscopic and analytical techniques, analysis of the size and size distribution of these nanosized vesicles is possible. However, due to the limited spatial resolution of conventional vibrational spectroscopy techniques, the chemical composition of individual nanosized liposome cannot be resolved. To address this limitation, we applied atomic force microscopy infrared spectroscopy (AFM-IR) to assess the chemical composition of individual liposomes encapsulating ciprofloxacin in dissolved and nanocrystalline form. Spatially resolved AFMIR spectra acquired from individual liposomes confirmed the presence of peaks related to N-H bending vibration, C-N stretching and symmetric and asymmetric vibration of the carboxyl group present in the ciprofloxacin. Our results further demonstrated the effectiveness of AFM-IR in differentiating the liposome containing ciprofloxacin in dissolved or nano-crystalline form. Spectra acquired from dissolved ciprofloxacin had peaks related to the ionised carboxyl group, i.e. at 1576 cm-1 and 1392 cm-1, which were either absent or far weaker in intensity in the spectra of liposomal sample containing ciprofloxacin nanocrystals. These findings are highly significant for pharmaceutical scientists to ascertain the stability and physicochemical composition of individual liposomes and will facilitate the design and development of liposomes with greater therapeutic benefits