Lung Delivery Research Articles

Zwitterionic Polymer-Functionalized Lipid Nanoparticles for the Nebulized Delivery of mRNA.

Lipid nanoparticles (LNPs) have great potential to enable inhaled delivery of mRNA to treat pulmonary diseases. However, this potential has been limited by the challenge of nebulizing the LNPs. Nebulization of LNPs can cause LNPs to aggregate and release encapsulated mRNA, limiting their delivery efficacy. To overcome this challenge, LNPs are developed whereby the PEG-lipid is fully replaced with a zwitterionic polymer (ZIP)-lipid conjugate to greatly enhance the nebulizer stability. LNPs formulated with ZIP-lipids (ZIP-LNPs) were stable to nebulization across a wide range of formulation parameters. The optimized ZIP-LNP formulation, containing reduced cholesterol content relative to traditional PEG-lipid LNPs, demonstrated improved inhaled mRNA delivery in both healthy and mucoobstructed mouse lungs. Repeat administration of the optimized ZIP-LNP formulation was well tolerated and did not result in pulmonary inflammation. This study demonstrates the potential of zwitterionic polymer-lipid conjugates for improving the performance of inhaled mRNA-LNP formulations.

Prediction of Transport and Deposition of Porous Particles in the Respiratory System Using Eulerian-Lagrangian Approach.

Deep lung delivery is crucial for respiratory disease treatment. Although nano and submicron particles exhibited a good deposition efficiency in deep regions of the lung, powder nonuniformity and particle agglomeration reduce their efficiency. Inhalation of porous particles (PPs) can overcome the mentioned challenges due to their larger size and low-density. The present study numerically investigates the deposition and penetration efficiency of orally inhaled PPs. A revised drag coefficient was implemented for PP transport. A realistic mouth-throat to the fifth generation of the lung was reconstructed from CT-scan images. A dilute suspension of uniformly distributed particles was considered at three inhalation flow rates (15, 30, and 45 L/min). Governing equations of the flow field and particle transport are solved using an Eulerian-Lagrangian approach. The results demonstrate that inhaling PPs significantly reduces the total and regional deposition of particles. There was also a critical porosity value under moderate and high inhalation flow rates for large PPs. Below this critical value, PP deposition efficiency substantially decreases. Additionally, it was also found that under low inhalation flow rates, the impact of porosity value is negligible. Almost 95% of the PPs penetrate the lower branches. These findings provide particle engineers and pharmaceutics with profound insight into developing novel inhalation techniques and drug delivery methods for deep lung delivery.

Advanced Spray-Dried Inhalable Microparticles/Nanoparticles of an Innovative Mitophagy Activator for Targeted Lung Delivery: Design, Comprehensive Characterization, Human Lung Cell Culture, and In Vitro Aerosol Dispersion Performance.

Urolithin A (UA) has demonstrated the ability to stimulate mitophagy and enhance mitochondrial and cellular health in skeletal muscles in humans after oral administration. It is hypothesized that targeted delivery of UA as inhaled dry powders to the lungs will enhance mitochondrial health through mitochondrial biogenesis. This study aimed to engineer inhalable excipient-free powders of UA as dry powder inhalers (DPIs) for targeted pulmonary delivery. The particles were designed by particle engineering from dilute organic solutions of UA using the state-of-the-art spray drying technology in a closed mode. Comprehensive physicochemical characterization and advanced microscopy techniques were conducted to examine phase behavior, molecular properties, and particle properties, which are necessary for the rational design of advanced pulmonary inhalation aerosols. Molecular fingerprinting was conducted by using attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy and Raman spectroscopy. Chemical imaging and mapping were conducted using confocal Raman microscopy (CRM) and IR microscopy. The advanced spray-dried (SD) excipient-free powders were successfully produced at different spraying pump feed rates and exhibited favorable molecular and particle properties. The excipient-free SD powders exhibited outstanding in vitro aerosol dispersion performance with an FDI-approved human DPI device (Neohaler) and correlated with the spray drying pump rate. In vitro, cell viability of various human pulmonary cells from different lung regions demonstrated biocompatibility and safety at different doses of UA. The transepithelial electrical resistance (TEER) assay shows that UA maintains cell membrane integrity and barrier tightness, indicating its potential for safe and effective localized drug delivery without long-term adverse effects. These results demonstrated that UA has favorable physicochemical and in vitro properties for inhalation and can be successfully engineered into excipient-free inhalable microparticles/nanoparticles as DPIs.

Inhalable Advanced Co-Spray Dried Microparticles/Nanoparticles of a Novel RhoA/Rho Kinase Inhibitor with Lung Surfactant Biomimetic Phospholipids for Targeted Lung Delivery.

Co-spray dried inhalable powder formulations of fasudil monohydrochloride salt (FMCS) and inhalable lung surfactant-based nanocarriers composed of synthetic phospholipids, zwitterionic DPPC (1,2-palmitoyl-sn-glycero-3-phosphocholine) and anionic DPPG (1,2-dipalmitoyl-sn-glycero-3-[phosphor-rac-1-glycerol]) sodium salt, were designed and optimized using organic solution advanced spray drying. FMCS can potentially be used for the treatment of various complex pulmonary diseases with this current work focusing on pulmonary arterial hypertension. Comprehensive physicochemical characterization, electron and optical microscopy imaging, thermal analysis, molecular fingerprinting spectroscopy, in vitro aerosol dispersion performance with human dry powder inhaler (DPI) devices, in vitro membrane permeation and drug release, and in vitro human cellular studies were conducted. Well-defined, small, and smooth nanoparticles/microparticles in the solid state were engineered at different molar ratios of FMCS/DPPC/DPPG (25:75, 50:50, and 75:25) and successfully produced as inhalable powders having the properties necessary for targeted pulmonary delivery as dry powder inhalers. In vitro aerosol performance demonstrated excellent aerosol dispersion with different DPI devices. The phospholipid bilayer biophysical properties were confirmed and retained following cospray drying. Sustained release of fasudil drug and in vitro biocompatibility were demonstrated on human lung cells from different airway regions.

Characterization of pediatric extrathoracic aerosol deposition with air-jet dry powder inhalers

The use of air-jet dry powder inhalers (DPIs) offers a number of advantages for the administration of pharmaceutical aerosols, including the ability to achieve highly efficient and potentially targeted aerosol delivery to the lungs of children using the oral or trans-nasal routes of administration. To better plan targeted lung delivery of pharmaceutical aerosols with these inhalers, more information is needed on the extrathoracic (ET) depositional loss in pediatric subjects when using relatively small (e.g., 0.5–2 μm) particles and including oral or nasal device interfaces. The objective of this study was to implement validated computational fluid dynamics (CFD) models to characterize ET depositional loss during mouth-throat (MT) and nose-throat (NT) aerosol administration to pediatric subjects (2–10 years old) using an air-jet DPI platform across a range of initial small-particle aerosol sizes (0.41–13.65 μm) and inhalation flow rates (8–20 L/min).A new CFD model focused on small-particle aerosol depositional loss in existing pediatric airway models was developed and validated with existing in vitro data. The validated CFD model was then used to characterize depositional loss in the MT and NT regions of children using particle sizes, flow rates and interfaces consistent with air-jet DPIs.Successful validation of the CFD model for small-particle aerosol deposition was achieved through enhanced resolution of the near-wall transport conditions. Existing non-dimensional parameters were used to produce high quality single-curve deposition efficiency correlations with r2 values in the range of 0.95–0.97. A new method for predicting realistic polydisperse aerosol deposition using the developed correlations and an equivalent monodisperse particle diameter was also introduced. In conclusion, the newly developed correlations will be useful in planning the lung delivery of next-generation inhaled medications, where achieving both low ET loss and targeted airway deposition, perhaps with excipient enhanced growth technology, are critical factors.

Poly(malic acid) Nanoconjugates of Pyrazinoic Acid for Lung Delivery in the Treatment of Tuberculosis.

Tuberculosis (TB) remains a major global infection, and TB treatments could be improved by site-specific targeting with delivery systems that allow tissue and cell uptake. To increase the drug concentration at the target sites following lung delivery, polymeric nanoconjugates based on biodegradable poly(malic acid) were designed. Pyrazinoic acid (POA), the active moiety of pyrazinamide─a first-line antituberculosis drug─was covalently bound to poly(malic acid) using a hydrophobic linker at mole ratios of 25%, 50%, and 75%. Three linkers, hexanediol, octanediol, and decanediol, were considered. Independently of the linker or ratio, all the conjugates were able to self-assemble, forming nanoconjugates (NCs) in water with 130-190 nm in diameter. Pyrazinoic acid could be released in a controlled manner without any burst release effect. Its kinetics can be adjusted by modifying the grafting ratio and linker length. No cytotoxicity was observed on RAW 264.7 macrophages up to ∼14 μg/mL of POA. In addition, the nanoconjugates were efficiently taken up by these cells over 5 h. Thanks to their high loading capacity and modulable release profiles, these nanoconjugates hold great promise for more effective treatment of tuberculosis.

Cross-Linked Thiolated Hydroxypropil-β-Cyclodextrin for Pulmonary Drug Delivery.

The effectiveness of cross-linked HP-β-CD-SH nanoparticles (HP-β-CD-SH-NP) was compared to transient nanoaggregates in enhancing the activity of dexamethasone (DMS) and olive leaf extracts (OLE). DMS, a poorly soluble drug commonly used in lung treatments, and OLE, known for its antioxidant properties, were chosen. Drug-loaded HP-β-CD-SH-NP were prepared and nebulized onto a lung epithelial Air-Liquid Interface (ALI) model, assessing drug permeation and activity. HP-β-CD-SH with 25% thiolation was synthesized via microwave reaction, forming 150 nm nanoaggregates and stabilized 400 nm HP-β-CD-SH-NP. All carriers showed good complexing ability with DMS and OLE and were biocompatible in the lung ALI model. HP-β-CD-SH promoted DMS absorption, while stabilized HP-β-CD-SH-NP protected against oxidative stress. HP-β-CD-SH is promising for lung delivery, especially as stabilized nanoaggregates, offering versatile administration for labile molecules like natural extracts.

Inhalable drug-loaded silk fibroin carriers for pulmonary drug delivery.

The design and development of engineered micro and nano-carriers offering superior therapeutic performance compared to traditional delivery forms, are crucial in pharmaceutical research. Aerosolization and inhalation of carriers with improved solubility/stability of insoluble drugs, has huge potential for targeted drug delivery (DD) in various pulmonary diseases. Indeed, dedicated carriers must meet specific dimensional rules for proper lung delivery. Particles between 2-10 μm in size are normally deposited in the tracheobronchial region, while particles of 0.5-2 μm may be properly deposited in the alveoli. In this work, we report the development of inhalable nanostructured carries made of a 'green' bio-inspired polymer from aqueous solutions, i.e. silk fibroin (SF), efficiently loaded with a hydrophobic drug, i.e. the thyroid hormone levothyroxine (L-T4), a drug for the treatment of idiopathic pulmonary fibrosis. The aim is to optimize a standard method for the synthesis of SF-based nanocarriers with controlled size and shape, where a fine control of their geometrical properties is aimed at efficiently controlling the pulmonary DD. L-T4 loaded SF particles were synthesized through a one-pot co-precipitation method. Optimized systems were determined by varying the chemo-physical parameters during the synthesis. Ethylenediaminetetraacetic acid (EDTA) was used to remove CaCO3 cores. The proposed synthesis routes have led to two SF structures, whose structural heterogeneity and nanostructured morphology have been demonstrated using fluorescence microscopy, DLS, SEM and EDX. Two systems with varying shape and size have been obtained: (i) a flat disk-like SF structure with an irregular surface and an in-plane length of about 1-2 μm; (ii) solid SF nanospheres, obtained using ethylene glycol as additive, showing two size populations (main diameters of 0.5 μm and 1.7 μm). Solid nanospherical systems, in particular, show a tendency to arrange into agglomerates that, when loosely bound into smaller particles, can facilitate the delivery at the alveoli. Both formulations exhibited similar drug loading efficiencies, evaluated by HPLC analysis. However, SF nanospheres showed greater in vitro drug release after 24 hours. The demonstrated control over the characteristics imparted to the proposed DD systems may be critical to select the most suitable size/shape to achieve high rates of delivery to the appropriate lung compartment.

Pharmacological and pre-clinical safety profile of rSIV.F/HN, a hybrid lentiviral vector for cystic fibrosis gene therapy.

Cystic fibrosis (CF) is caused by mutations in the CF Transmembrane Conductance Regulator (CFTR) gene. CFTR modulators offer significant improvements, but approximately 10% of patients remain nonresponsive or are intolerant. This study provides an analysis of rSIV.F/HN, a lentiviral vector optimized for lung delivery, including CFTR protein expression, functional correction of CFTR defects and genomic integration site analysis in preparation for a first-in-human clinical trial. Air-liquid interface cultures of primary human bronchial epithelial cells (HBEC) from CF patients (F508del/F508del), as well as a CFTR-deficient immortalized human lung epithelial cell line mimicking Class I (CFTR-null) homozygous mutations, were used to assess transduction efficiency. Quantification methods included a novel proximity ligation assay (PLA) for CFTR protein expression. For assessment of CFTR channel activity, Ussing chamber studies were conducted. The safety profile was assessed using integration site analysis and in vitro insertional mutagenesis studies. rSIV.F/HN expressed CFTR and restored CFTR-mediated chloride currents to physiological levels in primary F508del/F508del HBECs as well as in a Class I cells. In contrast, the latter could not be achieved by small-molecule CFTR modulators, underscoring the potential of gene therapy for this mutation class. Combination of rSIV.F/HN-CFTR with the potentiator ivacaftor showed a greater than additive effect. The genomic integration pattern showed no site predominance (frequency of occurrence ≤10%), and a low risk of insertional mutagenesis was observed in an in vitro immortalization assay. The results underscore rSIV.F/HN as a promising gene therapy vector for CF, providing a mutation-agnostic treatment option.

Development of a proliposomal pretomanid dry powder inhaler as a novel alternative approach for combating pulmonary tuberculosis

Multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) continue as public health concerns. Inhaled drug therapy for TB has substantial benefits in combating the causal agent of TB (Mycobacterium tuberculosis). Pretomanid is a promising candidate in an optional combined regimen for XDR-TB. Pretomanid has demonstrated high potency against M. tuberculosis in both the active and latent phases. Conventional spray drying was used to formulate pretomanid as dry powder inhalers (DPIs) for deep lung delivery using a proliposomal system with a trehalose coarse excipient to enhance the drug solubility. Co-spray drying with L-leucine protected hygroscopic trehalose in formulations and improved powder aerosolization. Higher amounts of L-leucine (40–50 % w/w) resulted in the formation of mesoporous particles with high percentages of drug content and entrapment efficiency. The aerosolized powders demonstrated both geometric and median aerodynamic diameters < 5 µm with > 90 % emitted dose and > 50 % fine particle fraction. Upon reconstitution in simulated physiological fluid, the proliposomes completely converted to liposomes, exhibiting suitable particle sizes (130–300 nm) with stable colloids and improving drug solubility, leading to higher drug dissolution compared to the drug alone. Inhalable pretomanid showed higher antimycobacterial activity than pretomanid alone. The formulations were safe for all broncho-epithelial cell lines and alveolar macrophages, thus indicating their potential suitability for DPIs targeting pulmonary TB.

Age-dependent changes in phagocytic activity: in vivo response of mouse pulmonary antigen presenting cells to direct lung delivery of charged PEGDA nanoparticles

BackgroundCurrent needle-based vaccination for respiratory viruses is ineffective at producing sufficient, long-lasting local immunity in the elderly. Direct pulmonary delivery to the resident local pulmonary immune cells can create long-term mucosal responses. However, criteria for drug vehicle design rules that can overcome age-specific changes in immune cell functions have yet to be established.ResultsHere, in vivo charge-based nanoparticle (NP) uptake was compared in mice of two age groups (2- and 16-months) within the four notable pulmonary antigen presenting cell (APC) populations: alveolar macrophages (AM), interstitial macrophages (IM), CD103+ dendritic cells (DCs), and CD11b+ DCs. Both macrophage populations exhibited preferential uptake of anionic nanoparticles but showed inverse rates of phagocytosis between the AM and IM populations across age. DC populations demonstrated preferential uptake of cationic nanoparticles, which remarkably did not significantly change in the aged group. Further characterization of cell phenotypes post-NP internalization demonstrated unique surface marker expression and activation levels for each APC population, showcasing heightened DC inflammatory response to NP delivery in the aged group.ConclusionThe age of mice demonstrated significant preferences in the charge-based NP uptake in APCs that differed greatly between macrophages and DCs. Carefully balance of the targeting and activation of specific types of pulmonary APCs will be critical to produce efficient, age-based vaccines for the growing elderly population.Graphical

Development of a New Dry Powder Aerosol Synthetic Lung Surfactant Product for Neonatal Respiratory Distress Syndrome (RDS) – Part I: In Vitro Testing and Characterization

PurposeImproving the deep lung delivery of aerosol surfactant therapy (AST) with a dry powder formulation may enable significant reductions in dose while providing improved efficacy. The objective of Part I of this two-part study was to present the development of a new dry powder aerosol synthetic lung surfactant (SLS) product and to characterize performance based on aerosol formation and realistic in vitro airway testing leading to aerosol delivery recommendations for subsequent in vivo animal model experiments.MethodsA new micrometer-sized SLS excipient enhanced growth (EEG) dry powder formulation was produced via spray drying and aerosolized using a positive-pressure air-jet dry powder inhaler (DPI) intended for aerosol delivery directly to intubated infants with respiratory distress syndrome (RDS) or infant-size test animals.ResultsThe best-case design (D2) of the air-jet DPI was capable of high emitted dose (> 80% of loaded) and formed a < 2 µm mass median aerodynamic diameter (MMAD) aerosol, but was limited to ≤ 20 mg mass loadings. Testing with a realistic in vitro rabbit model indicated that over half of the loaded dose could penetrate into the lower lung regions. Using the characterization data, a dose delivery protocol was designed in which a 60 mg total loaded dose would be administered and deliver an approximate lung dose of 14.7–17.7 mg phospholipids/kg with a total aerosol delivery period < 5 min.ConclusionsA high-efficiency aerosol SLS product was designed and tested that may enable an order of magnitude reduction in administered phospholipid dose, and provide rapid aerosol administration to infants with RDS.

In Vitro Comparison of Aerosol Delivery in High-Frequency Assisted Airway Clearance Devices With Integrated Nebulizers.

High-frequency assisted airway clearance systems combine positive expiratory pressure or oscillatory positive airway pressure with integrated nebulizers to improve the delivery of aerosols and assist with airway clearance. This aerosol study evaluated lung delivery efficiency during positive expiratory pressure and oscillatory positive airway pressure therapy of 2 high-frequency assisted airway clearance/nebulizer systems. Aerosol delivery was evaluated during positive expiratory pressure therapy of 10 cm H2O and oscillatory positive airway pressure therapy of 20 cm H2O with the BiWaze Clear and the Volara high-frequency assisted airway clearance/nebulizer systems. The handset and nebulizer were attached to an anatomic upper-airway model via a mouthpiece and placed into a plethysmograph. A tracheal filter was placed to capture the inhaled aerosol. A vacuum filter entrained fugitive aerosols from the plethysmograph. After nebulization of technetium in 3.0 mL normal saline solution, the components were scanned by using scintigraphy and the decay-corrected radiation counts were referenced to the initial nebulizer technetium charges. Aerosol delivery during positive expiratory pressure therapy of 10 cm H2O resulted in higher lung deposition with the BiWaze Clear versus the Volara (28 vs 6.2%; P < .001; 95% CI 16.5-27.7), and higher fugitive losses (23.7 vs 2.8%; P = .004) and nebulizer losses (55 vs 3.3%; P < .001) with the Volara than with the BiWaze Clear. Aerosol delivery during oscillatory positive airway pressure of 20 cm H2O resulted in a higher lung deposition with the BiWaze Clear versus the Volara (16.3 vs 7.3%; P = .005; 95% CI 3.3-15) and higher fugitive (22.3 vs 3.8%; P = .02) and nebulizer (58.8 vs 7.2%; P = .004) losses with the Volara. There were no differences at the other locations during testing. The BiWaze Clear system showed greater delivery efficiency than did the Volara during positive expiratory pressure and oscillatory positive airway pressure. The high residual nebulizer dose and fugitive aerosol losses through the handset leak valve contributed to the lower delivery efficiency observed with the Volara. The nebulizer type, circuit design, and handset are important factors when targeting effective aerosol delivery to the lungs with high-frequency assisted airway clearance therapy.

Oral inhalation of dacomitinib nanocarriers as a therapeutic strategy for non-small cell lung cancer.

Background: Development of an inhalable nanoformulation of dacomitinib (DMB) encapsulated in poly-(lactic-co-glycolic acid) nanoparticles (NPs) to improve solubility, facilitate direct lung delivery and overcome the systemic adverse effects.Methods: DMB-loaded poly-(lactic-co-glycolic acid) NPs were prepared using solvent evaporation and characterized for particle size, polydispersity index and zeta-potential. The NPs were evaluated for invitro drug release, aerosolization performance and in vitro efficacy studies.Results: The NPs showed excellent particle characteristics and displayed a cumulative release of ∼40% in 5days. The NPs demonstrated a mass median aerodynamic diameter of ∼3 μm and fine particle fraction of ∼80%. Further, in vitro cell culture studies showed improved cytotoxic potential of DMB-loaded NPs compared with free drug.Conclusion: The study underscores the potential of DMB-loaded NPs as a viable approach for non-small cell lung cancer treatment.

Quercetin Solid Lipid Microparticle Stability and Deposition in Rat Lungs: A Study of Surfactant Effect

This study aims to determine the effect of surfactant concentration on Quercetin SLM as a potential carrier of respiratory diseases, especially ones resulting from oxidative stress injury. Quercetin is a natural antioxidant with high activity. SLM was formed with 5% compritol 888 ATO as a lipid and different concentrations of poloxamer 188 as a surfactant. SLM was produced by a combination of emulsification and sonication involving freeze drying. SLM is characterized by organolepsis, morphology, yield, particle size, drug loading, and entrapment efficiency. The antioxidant activity of quercetin SLMs was tested using the ABTS method. SLMs are characterized as having round and smooth morphology, high yield (F1 88.53%; F2 91.44%; F3 92.87%); particle size (F1 1.81 um; F2 1.90 um; F3 1.94 um); high drug loading (F1 15.96%; F2 13.74%; F3 13.19%); and high entrapment efficiency (F1 96.53%; F2 87.94%; F3 87.48%). Increasing surfactant concentration did not produce a significant difference between formulas. Quercetin SLM showed high antioxidant activity (Quercetin 94.43%; F1 94.35%; F2 94.36%; F3 94.37%). SLM was stable at storage temperatures between 25°C and 40°C. The effect of surfactant can be seen on particle size, drug loading, and entrapment efficiency at 40°C. Results of in vivo deposition study indicated that all SLM formulas were able to deliver quercetin to the lungs. Increasing the concentration of surfactant in Quercetin SLMs made no difference to the lung deposition as confirmed by observations conducted at 1 hour and 4 hours. Quercetin SLM has the potential for lung delivery by inhalation.

Preparation and Evaluation of Inhalable Microparticles with Improved Aerodynamic Performance and Dispersibility Using L-Leucine and Hot-Melt Extrusion.

Dry-powder inhalers (DPIs) are valued for their stability but formulating them is challenging due to powder aggregation and limited flowability, which affects drug delivery and uniformity. In this study, the incorporation of L-leucine (LEU) into hot-melt extrusion (HME) was proposed to enhance dispersibility while simultaneously maintaining the high aerodynamic performance of inhalable microparticles. This study explored using LEU in HME to improve dispersibility and maintain the high aerodynamic performance of inhalable microparticles. Formulations with crystalline itraconazole (ITZ) and LEU were made via co-jet milling and HME followed by jet milling. The LEU ratio varied, comparing solubility, homogenization, and aerodynamic performance enhancements. In HME, ITZ solubility increased, and crystallinity decreased. Higher LEU ratios in HME formulations reduced the contact angle, enhancing mass median aerodynamic diameter (MMAD) size and aerodynamic performance synergistically. Achieving a maximum extra fine particle fraction of 33.68 ± 1.31% enabled stable deep lung delivery. This study shows that HME combined with LEU effectively produces inhalable particles, which is promising for improved drug dispersion and delivery.

The study of galactomannans with different molecular weights and their ability to form microparticles suitable for pulmonary delivery

The influence of locust bean gum (LBG) galactomannans (GMs) molecular weight (Mw) to assemble microparticulate systems was evaluated, and carriers for deep lung delivery were developed. A commercial batch of LBG with a mannose/galactose (M/G) ratio of 2.4 (batch 1) was used to study the influence of different microwave partial acid hydrolysis conditions on carbohydrate composition, glycosidic linkages, and aqueous solutions viscosity. The microwave treatment did not affect the composition, presenting 4-Man (36–42 %), 4,6-Man (27–35 %), and T-Gal (24–25 %) as the main glycosidic linkages. Depolymerization led to a viscosity reduction (≤0.005 Pa·s) with no major impact on polysaccharide debranching. The structural composition of the LBG galactomannans were further elucidated with sequence-specific proteins using carbohydrate microarray technologies. A second batch of LBG (M/G 3.3) was used to study the impact of GMs with different Mw on microparticle assembling, characteristics, and insulin release kinetics. The low-Mw GMs microparticles led to a faster release (20 min) than the higher-Mw (40 min) ones, impacting the release kinetics. All microparticles exhibited a safety profile to cells of the respiratory tract. However, only the higher-Mw GMs allowed the assembly of microparticles with sizes suitable for this type of administration.

In silico investigation of inhalation condition impacts on hygroscopic growth and deposition of salbutamol sulphate in human airways

The objective of this study is to explore the transport, size growth, and deposition of Salbutamol Sulphate (SS) using Computational Fluid Dynamics (CFD). A CT-based realistic model of human airways from the oral cavity to the 5th generation of the lung was utilized as the computational domain. Four Test Cases (TC) with varying temperature and relative humidity (RH) under two inspiratory waveforms were considered to completely evaluate the impact of inhalation conditions on particle growth. Salbutamol Sulphate (SS) is a β2-adrenergic agonist and has been extensively used for asthma treatment. A monodispersed distribution of SS particles with an initial diameter of 167 nm was considered at the mouth inlet based on pharmaceutical data. Results indicated that inhalation of saturated/supersaturated air (RH>100%) leads to significant hygroscopic growth of SS particles with a factor of 10. In addition, the deposition efficiency of SS particles under the Quick and Deep (QD) inhalation profile was enhanced as the flow temperature and humidity increased. However, the implementation of Slow and Deep (SD) inspiratory waveform revealed that the same particle size growth is achieved in the respiratory system with lower deposition efficiency in the mouth-throat (less than 3%) and tracheobronchial airway (less than 2.18%). For the escaped particles form the right lung, in the SD waveform under TC 3, the maximum particle size distribution was for 600 nm particles with 25% probability. In the left lung, 30% of the particles were increased up to 950 nm in size. For the QD waveform in TC 3 and TC4, the most frequent particles were 800 nm with 36% probability. This holds practical significance in the context of deep lung delivery for asthmatic patients with enhanced deposition efficiency and large particle size. The findings of the present study can contribute to the development of targeted drug delivery strategies for the treatment of pulmonary diseases using hygroscopic dry powder formulations.

Design of experiments using box behnken design in the development, characterization, mathematical modeling, and evaluation of lung targeted nebulized antiviral camostat mesylate loaded pegylated nanosuspension product

Box Behnken Design is a factorial statistical approach that helps to overcome limitations associated with time-consuming full factorial experimental design. This study used the Box Behnken design and Design of Experiments (DoE) to develop the nebulized antiviral Camostat Mesylate loaded pegylated nanoliposomal suspension product with the target profile specifications of particle size <250 nm, polydispersity index (PDI) between 0.1 and 0.3, and zeta potential (ZP) of 0±5 mV, % entrapment efficiency (%EE) between 40%w/v to 70 % w/v and % drug loading between 20%w/w to 50%w/w. Box Behnken design with a robust standard least squares model was employed to develop the pharma product. Camostat Mesylate loaded pegylated nanoliposomal suspension was developed using the scalable ethanol injection method. The three independent factors at three levels were concentrations of the lipids employed to develop the formulation. Applying holistic QbD during formulation development using Design of Experiments (DoE) ensures a science-based, risk-managed approach to developing high-quality pharmaceutical products. The novelty in this investigation is that, for the first time, nebulized antiviral Camostat mesylate-loaded Nanoliposomes with a size of <250 nm, size <250 nm, polydispersity index (PDI) between 0.1 and 0.3, and zeta potential (ZP) of 0±5 mV, % entrapment efficiency (%EE) between 40%w/v to 70 % w/v and % drug loading between 20%w/w to 50%w/w were developed using DoE, Box Behnken Design, mathematical modeling and Response Surface Methodology.Furthermore, the Camostat mesylate-loaded Nanoliposomes were evaluated for inhibiting SARS-CoV-2 injection. Based on our findings, we kept other concentrations, i.e., Camostat Mesylate concentration at 2.25 mg/mL, 0.9%w/v NaCl, and 90%v/v ethanol and process parameters, namely, stirring speed, stirring time, ethanol addition rate, syringe diameter constant. From the mathematical modeling and Response Surface Methodology, we observed that the particle size of the nanosuspension increased with an increase in concentrations of DPPC and cholesterol. At the same time, increasing concentrations of DPPC increased the % entrapment efficiency and % drug loading; an increase in the concentration of Cholesterol led to a decrease in both those parameters. DoE successfully developed Camostat Mesylate loaded pegylated nanoliposomes with a particle size of 188 ± 0.75 nm, 75.47 ± 5.87 % entrapment of Camostat Mesylate and 49.65 ± 3.86 % drug load of Camostat Mesylate in liposomal product using the ethanol-injection method meeting target profile. The in vitro extended release of Camo from the Camo-pegNLs was observed in 24 h. The nebulized Camo-pegNL product showed MMAD of 4.01 ± 0.21 μm, GSD of 1.34 ± 0.03, and %FPF of 29.53 ± 6.5 %. In conclusion, integrating nebulized product development, Box Behnken Design, enabled the successful development of nebulized Camostat Mesylate loaded pegylated nanoliposomal suspension product, demonstrating their suitability for lung delivery of the antiviral drug.

Cellular Distribution and Intracellular Localization of Different Sizes of Fluorescent Thiol-Organosilica Particles in Mouse Lungs.

We investigated the distribution of intratracheally administered thiol-organosilica (thiol-OS) particles in mouse lungs. Toward this end, single doses of thiol-OS particles containing fluorescein (140 nm in diameter) (F140) and rhodamine B (Rh) (Rh160, Rh280, Rh420, Rh640, and Rh1630 with diameters of 160, 280, 420, 640, and 1630 nm, respectively) were administered. After 24 h, fluorescence imaging revealed homogeneous fluorescence with a patchier pattern on the lung surface and no difference among the six particle sizes. Simultaneous dual administration of Rh and F140 particles did not reveal any size-dependent differences in the lung surface fluorescence. Fluorescence microscopy of the lung sections revealed a similar tissue distribution in the fluorescent areas of Rhs and F140. Some fluorescent areas showed one type of particle fluorescence or only one fluorescence. Cellular distribution of particles was observed in bronchoalveolar lavage cells and lung sections under a high magnification, and correlative light and electron microscopy revealed large cells with fluorescence corresponding to both particle types and small cells with fluorescence of individual particle types, indicating a cell-subset-dependent particle size effect. Rh280, Rh420, and Rh640 exhibited significant size effects and were taken up by alveolar macrophages. Extracellular particles were observed, indicating that saturation exceeded the particle dose threshold in the alveoli. F140 taken up by small and large macrophages colocalized with CD68, CD11c, and CD11b and correlated with CD11c. The size effect, intracellular localization, and extracellular distribution of particles provide insights into lung and systemic drug delivery.