Of mice and men: Correlation and clinical relevance of animal models and non‐clinical tests for inhaled pharmaceuticals
Daniela Traini1,2
1Respiratory Technology, Woolcock Institute of Medical Research, Sydney, NSW 2037, Australia
2Macquarie Medical School, Department of Biological Sciences, Faculty of Medicine, Health and Human Sciences, Macquarie University, NSW 2109, Australia
Summary: With the new FDA Modernization Act of 2022, the agency now has the option to approve drugs and biomolecules that are tested in only non‐clinical studies, including those that use cells and lab‐grown tissues, such as organs on a chip, or computer models, before being tested in human clinical trials. Although this is probably going to be a slow change and not a tsunami that will transform the drug approval process overnight, now is probably the time to start reviewing the status quo on what are the reasons behind current inhalation drug development preclinical models, what alternative models are available and how the research community is considering this field moving forward. For many years, we have been stuck in a particular conceptual mode of doing research, but with this amendment in policy change will arise as the weight of the evidence for alternative tools becomes more robust and uniformly accepted and validated. In this paper, a review of current and novel research on this topic will be presented, and the author provide possible future directions in this exciting field.
Inhaler Technique mastery; focus on LEARNING rather than TRAINING
Mark Sanders1 & Darragh Murnane1
1University of Hertfordshire, College Lane, Hatfield, AL10 9AB, UK
Summary: Inhaler technique errors are a widespread problem across all countries and all types of inhalers. Some patients receive training, but many do not. Where training is provided it is often only simple verbal counselling. The emphasis, in numerous guidance documents has been on training, not on learning. By focusing on learning we have identified some principles of Educational Theory that relate to healthcare. We have undertaken an audit of inhaler training tools and reviewed related clinical evidence to examine how widely the principles are applied, and what benefits are derived. Learning styles differ among individual patients, some people favour stimuli that are visual, others aural, or reading, or kinesthetic, or a mix of these modalities. Additionally, positive reinforcement can shape behaviour and real‐time feedback can help guide actions. Most inhaler training tools only engage a limited range of learning stimuli, more can be done. Re‐design of inhalers and inhaler training tools to incorporate proven principles of Educational Theory could help to enable more patients to learn to use their devices correctly, and to reduce the incidence of inhaler technique errors.
Digital technologies to improve adherence: Limitations and opportunities
Rob Horne
Centre for Behavioural Medicine, UCL School of Pharmacy, University College London
Summary: The proliferation of smart digital technologies has the potential to improve adherence and reduce the global burden of respiratory disease. Traditionally, such technologies have been used to address practical barriers to adherence, making treatment engagement as easy as possible. Although helpful, this has a limited effect (the so‐called ‘app trap’) because much of non‐adherence is intentional: the patient is able to adhere but does not want to. For adherence support to be most effective, it should address barriers to both motivation (I don't want to) and ability (I can't) to adhere. In this talk, we will apply evidence from behavioural science to show how people make decisions about treatment. The Perceptions and Practicalities ApproachTM (PAPA), a framework endorsed by NICE, is a way to understand the psychology of adherence. To improve adherence, interventions need to address perceptual barriers (e.g., patient beliefs, preferences and emotions) influencing motivation and practical barriers (e.g., capacity and resources) influencing ability to adhere to treatment. This presentation will show how PAPATM‐based digital and non‐digital interventions have significantly increased patient beliefs about the need for their medicine, reduced patient concerns about taking their medicine and increased medication adherence. We will also apply the 3C's of intervention design: Content, Channel and Context. Digital technologies offer an important Channel for the delivery of adherence support, but we need to couple technological expertise with behavioural science to optimise content (what apps say and do) and Context (the environment in which they are delivered).
European Respiratory Society Clinical Research Collaboration CONNECT. Moving multiple digital innovations towards connected respiratory care: Addressing the over‐arching challenges of whole systems implementation
Hilary Pinnock
The University of Edinburgh, Doorway 3, Medical School, Teviot Place, Edinburgh EH8 9AG, UK
Summary: Digital health is a priority for the European Respiratory Society (ERS) that can offer patients tailored, seamless (self‐)management support including remote access to clinical advice; enabling clinicians to monitor and tailor care; providing healthcare systems with data to target resources and monitoring/intervening in health events. Accelerated by the pandemic, the focus is shifting from pilot and effectiveness studies in specific clinical contexts to sustainable implementation of digital healthcare. However, successful implementation is complex requiring consideration of implications for patients, professionals and organisations, as well as an understanding of the political, regulatory, socio‐economic context. The Clinical Research Collaboration CONNECT, launched by ERS in April 2023, has developed a global multidisciplinary network of 900 colleagues in 80 countries focussed on implementing digital respiratory healthcare.
We aim to bridge the gap between disease and/or location‐focussed digital initiatives and implementation of sustainable, equitable, connected digital healthcare in routine clinical respiratory practice within European (and global) healthcare systems. CONNECT will enable knowledge exchange, create a repository of available respiratory technologies, promote standardised approaches to implementation research in digital healthcare, publish position paper(s) on barriers and enablers to deployment at scale, and plan research that will inform, develop and evaluate implementation of digital respiratory healthcare.
Inhalable RNA formulations based on pulmonary surfactant and repurposed cationic amphiphilic drugs
Koen Raemdonck
Ghent Research Group on Nanomedicines, Laboratory of General Biochemistry and Physical Pharmacy, Department of Pharmaceutics, Faculty of Pharmaceutical Sciences, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium.
Summary: The clinical approval of the first RNA interference (RNAi)‐based medicinal product Onpattro® (patisiran) in 2018 and the more recent clinical success of the SARS‐CoV‐2 messenger RNA (mRNA)‐based vaccines (Comirnaty® and Spikevax®) underscore the potential of RNA‐based drugs. To be functional, RNA drugs require delivery into the cytosol of target cells. Lipid nanoparticles (LNPs) have become a well‐established platform to package, protect and deliver various nucleic acid cargos. The lung is increasingly recognized as an attractive target organ for RNA delivery. However, in spite of many advantages of RNA inhalation therapy, to date no innovative RNA formulations are available for application in the lung. Moreover, the intracellular delivery efficiency of state‐of‐the‐art LNPs remains relatively low. This presentation will describe the repurposing of two distinct cationic amphiphiles, i.e. both low molecular weight cationic amphiphilic drugs (CADs) as well as the lung‐related surfactant protein B (SP‐B), to improve cellular delivery of RNA drugs for inhalation therapy.
Formulation and Chemistry Strategies to Overcome Pulmonary Drug Delivery Barriers
David Cipolla1, Adam J. Plaunt1, Tam L. Nguyen1, Michel R. Corboz1 & Vladimir S. Malinin1
1Insmed Incorporated, 700 US Highway 202/206, Bridgewater, NJ 08807, USA
Summary: The Drug Delivery to the Lung audience is cognizant of the challenges in developing inhaled therapeutics to treat lung disease. While there has been much focus on the optimization of the formulation and device technologies to better target delivery of a therapeutic to the lung, this presentation will discuss the biological barriers to pulmonary drug delivery and provide case studies of relatively novel formulation and chemistry strategies that can be utilized to overcome those barriers. The formulation and chemistry technologies include the use of nanoparticle formulations, including liposomes, as well as prodrug strategies to modify the disposition in the lung.
Enoximone and cyclodextrins: novel formulations for the treatment of pulmonary pathologies
C. Migone1, B. Grassiri1, L. Vizzoni1,2, Y. Zambito1, A. M. Healy2,3, C. Ehrhardt2,3, P. Roncucci4, B. Ferro4 & A. M. Piras1
1Department of Pharmacy, University of Pisa, Via Bonanno Pisano 33, Pisa/56126, Italy.
2Department of Life Sciences, University of Siena, Via P.A. Mattioli 4, Siena, 53100, Italy.
3SSPC, The Science Foundation Ireland Research Centre for Pharmaceuticals, Ireland.
4School of Pharmacy and Pharmaceutical Sciences, Trinity College Dublin, Dublin 2, Ireland.
5Departments of Anesthesia and Critical Care, Spedali Riuniti Livorno Estav Nordovest, Livorno/57121, Italy
Summary: Enoximone (Enox), is a phosphodiesterase 3 inhibitor (PD3), available as intravenous formulation (i.v.‐Enox), named Perfan®. Recently, i.v‐ENOX showed promising clinical results in the treatment of acute respiratory distress syndrome caused by COVID‐19. However, i.v.‐Enox has a pH of 12 and forms crystals in the ampole. The aim of this study was to develop new formulations suitable for pulmonary administration of Enox and to improve the drug's lung bioactivity. Firstly, a solution for nebulisation, based on Enox and hydroxypropyl‐β‐cyclodextrin (HPβCD) as a stabilising agent was developed (HPβCD/Enox) and characterised in terms of aerodynamic particle size distribution and biopharmaceutical characteristics. The evaluation of generated aerosol indicated a better peripheral lung distribution of Enox/HPβCD compared to i.v‐Enox. Biological evaluations showed that Enox has direct effect on human lung epithelial NCI‐H441 cells increasing intracellular cAMP levels, providing protection from oxidative stress when administered in presence of cyclodextrin. The second part of the work concerned the development of a powder formulation for Enox administration via dry powder inhaler (DPI). Spray dried formulations were lactose‐free and contained β‐cyclodextrin (βCD) derivatives to ensure Enox water solubility. Three cyclodextrins showing higher degrees of complex formation compared with HPβCD were evaluated: βCD, methyl‐β‐cyclodextrin (MβCD) and sulfobutyl ether β‐cyclodextrin (SBEβCD). In vitro deposition profiles were determined using a Next Generation Impactor (NGI) and indicated suitability for pulmonary drug delivery. Biopharmaceutical studies revealed that the delivery of Enox by DPI was particularly effective in increasing intracellular cAMP levels. Concluding, DPI formulations could provide an excellent starting point to improve Enox pulmonary bioactivity.
Plume Geometry: examination of processing schemes and inter‐analyst variability…We Can Do Better!
Christopher J. Gruenloh, Karen M. Schmidt, Jenna C. Caragiulo and Karina S. McKinney
PPD, Part of Thermo Fisher Scientific, 8556 Research Way, Middleton, WI 53562, USA
Summary: Spray pattern (SP) and plume geometry (PG) are spray characterization techniques used during the development of pressurized metered‐dose inhalers (pMDIs) and nasal sprays. SP is employed for routine quality control testing and both tests are required to evaluate test and innovator products in the generic approval pathway. PG is generally thought to not accurately reflect the performance of either device or formulation and is perceived as variable and subjective. One also needs to recognize as well as reconcile differences in the performance of SP and PG testing with a time‐averaged image submitted for processing the former whereas a single “snapshot” image is typically used for the latter. While today's SP test has become much less subjective through use of images captured by high‐speed cameras with laser light illumination and automated software processing routines, questions and perceptions remain for PG, where the analyst must select the frame processed, the spray angle origin and the placement of “arms” to indicate the outer limits of the plume. The objective of this study was to examine the repeatability of PG measurements across a series of 4 commercially available pMDI products with 3 analysts processing. Using the same data sets, the impacts of time‐averaging vs. snapshot images as well as different techniques to define the origin and outer edges of the plume were investigated. Data indicates that both inter‐analyst and product variability in PG processing can be greatly reduced through use of a time‐averaged image with an intensity‐based methodology to define the plume's outer edges.
A Multi‐Physics Theoretical and CFD Approach, To Predict Low‐GWP pMDI Spray Characteristics, Inside USP‐IP Geometry
1Kindeva Drug Delivery Limited, Derby Road, Loughborough, Leicestershire, LE11 5SF, UK
2Wolfson School of MEME, Loughborough University, Ashby Road, Loughborough, LE11 3TU, UK
Summary: This paper presents the development of a comprehensive mathematical model aimed at predicting pressurized metered‐dose inhaler (pMDI) unwanted drug deposition, using low global warming potential (GWP) propellant. The model incorporates various factors such as the geometric dimensions of pMDI's actuator/valve and the properties of the formulation such as propellant, excipient, and drug load. The model successfully predicts the drug deposition on the actuator and the USP‐IP, and, hence, the delivered fraction to the NGI/ACI. These predictions are validated through laboratory testing.
Steady or not? Using realistic, transient aerosol parameters to predict lung deposition
Maximilian J. Grill1, Benjamin Heine2, Jakob Richter1, Carolin Hein2, Johann Dietsche2, Amed Njoya2, Karl‐Robert Wichmann1, Jonas Biehler1, Kei W. Müller1
Summary: The output of nebulizers is commonly assumed to be constant over the breathing cycle. Accordingly, time‐averaged values are used to estimate aerosol deposition in the lungs. However, recent measurements show that especially nebulizers with aerosol storage chamber yield a highly unsteady aerosol output rate (uAOR), the so‐called “aerosol bolus”. Here, we developed the first method to measure droplet sizes over a breathing cycle, thereby extending transient aerosol characterization. We studied two nebulizers, one with (PARI eFlow) and one without (Philips InnoSpire Go) storage chamber. For the eFlow, the peak in uAOR was 320% of what is continuously produced by the aerosol head, compared to only 128% for the InnoSpire Go, clearly demonstrating different levels of unsteadiness. In addition, both systems exhibited a substantial change in droplet sizes over the breathing cycle. Because uAOR and droplet sizes are known to have a strong influence on lung deposition, we used measured transient parameter values as input for a recently proposed and validated, high‐fidelity in silico model for pulmonary drug delivery. Compared to the InnoSpire Go, simulation results for the eFlow indicated a 28% reduction in exhalation losses, a 58% reduction of deposition in the upper airways, and a 3‐fold increase in aerosol mass being deposited in the alveolar region in each breathing cycle. This outcome can be explained by finer droplet sizes and the bolus effect, i.e., the availability of more aerosol mass both in total and especially early in the inhalation. As a result, the eFlow enables much higher overall efficiency and much shorter treatment times.
Reducing Time Requirements for Morphologically Directed Raman Spectroscopy Based In Vitro Bioequivalence Studies via Advanced Optimisation of Analysis Methods
Benjamin Stafford1, Lily Richards1, Mark Parry1 & Mervin Ramjeeawon1
1Intertek Melbourn, Unit 4, Anglian Business Park, Royston, Hertfordshire, SG8 5TW, United Kingdom
Summary:In vitro only bioequivalence (IVBE) studies have formed part of increasing numbers of abbreviated new drug applications (ANDAs) for generic nasal drug products (OINDPs) over the past decade. Morphologically directed Raman spectroscopy (MDRS) can selectively measure the particle size and morphology of separate species within multi‐component blends and suspensions, allowing it to characterise the size of a drug, and infer the dissolution characteristics to a level where clinical endpoint studies may be entirely bypassed when demonstrating bioequivalence for certain products. This capability has resulted in high demand from generics developers for MDRS to be built into their testing programs. However, a typical MDRS measurement takes many hours to complete, such that in most cases only one sample per day, per system is analysed. Compared to other in vitro techniques, this limitation leads to unfavourably large time requirements for full IVBE studies which usually mandate a minimum of 60 replicates be performed. This study sought to investigate if the MDRS time requirements for an IVBE could feasibly be reduced whilst still maintaining confidence in the final outcome. An MDRS method to measure the size of API particles within mometasone furoate monohydrate nasal suspensions was optimised such that microscopy‐only analysis could selectively measure a PSD of the API, which was proved to be successful by later applying Raman chemical identifications. This outcome presents an opportunity to reduce the number of full MDRS replicates that need to be performed during an IVBE, allowing for a large reduction in time requirements. The remainder would be comprised of faster microscopy‐only based analysis, for which the high degree of API selectivity and agreement with MDRS has been demonstrated.
Development of pulmonary formulations with gallium siderophores against Aspergillus fumigatus lung infections
B. Grassiri1,2, M. Piatek3,4, L. More O'Ferrall3,5, S. Esin6, J. A. Sake2, G. Batoni6, C. Ehrhardt2, D. M. Griffith3,5, K. Kavanagh3,4, A. M. Piras1, A. M. Healy2,3
1University of Pisa, Department of Pharmacy, 56126 Pisa, Italy
2School of Pharmacy and Pharmaceutical Sciences, Trinity College Dublin, Dublin 2, Ireland
3SSPC, The Science Foundation Ireland Research Centre for Pharmaceuticals, Ireland
4Department of Biology, Maynooth University, Maynooth, Ireland
5Department of Chemistry, Royal College of Surgeons in Ireland, Dublin 2, Ireland
6Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56123, Pisa, Italy
Summary: The lack of available drugs for the treatment of fungal infections, along with the emergence of drug‐resistant strains, results in millions of deaths each year [1]. The World Health Organization has categorised Aspergillus fumigatus (A. fumigatus) in the critical priority group of fungal pathogens associated with serious risk of mortality and morbidity in humans. Pulmonary aspergillosis is particularly severe in patients with cystic fibrosis (CF) in which it is clearly associated with airway damage and lung function decline. In this work, the novel gallium‐siderophore complex, GaS1, was selected as the most active against A. fumigatus (ATCC strain 26933), while having low toxicity in NCI‐H441 distal lung epithelial cells. A formulation study was performed, resulting in a solution for nebulisation and three different dry powder inhaler (DPI) formulations, prepared by spray‐drying GaS1 with different concentrations of L‐leucine. Physicochemical characterisation of all the formulations was undertaken (i.e. SEM, PXRD, laser diffraction for the powders, and osmolality for the solution) and their aerodynamic deposition profiles were determined using the next generation impactor (NGI). All formulations showed suitable aerodynamic characteristics for GaS1 delivery to the lungs. Ultimately, the GaS1 based solution for nebulisation (namely GaS1‐sol) was assessed against clinical lung isolates of A. fumigatus within an in vitro lung infection model. This model was also employed to replicate fungal infections in the context of cystic fibrosis (CF) airway conditions, utilizing clinically isolated conidia, the NCI‐H441 cell line, and an artificial sputum medium (ASM). A marked dose dependent antifungal activity was observed, confirming the future potential lung delivery of gallium‐siderophores for fungal infection treatments.
Predicting Systemic Exposure to Inhaled Drugs using a Simplified Regional Deposition and Pharmacokinetic Method
Scott Tavernini1, Dino J. Farina2, Warren H. Finlay1 & Andrew R. Martin1
1Aerosol Research Lab of Alberta, 116 Street and 85 Avenue, Edmonton, T6G 2R3, Canada
Summary: Alternative in vitro characterization techniques have received significant attention recently due to the resource burden associated with cascade impactor measurements. One such alternative method uses specialized filters to enable direct measurement of expected tracheobronchial and alveolar doses of test aerosol penetrating an extrathoracic airway. Use of these regional deposition estimates as pharmacokinetic (PK) model inputs has not yet been explored. A PK model using two lung compartments was built to compute systemic exposure to budesonide based on previously measured in vitro deposition. The simplified model was compared to estimates arising from cascade impactor measurements and detailed modelling of deposition and disposition on a generational basis. Selection of an appropriate mucociliary clearance rate for the simplified model was achieved by matching overall tracheobronchial clearance between the two models. Despite the different methodologies, predicted PK profiles were in very close agreement. This demonstrates the ability to estimate systemic exposure to a compound of interest using fully in vitro regional deposition estimates.
Enhanced Lung Delivery of mRNA Using Nebulized Lipid Nanoparticles
1Hovione FarmaCiencia S.A., R&D Inhalation & Advanced Drug Delivery, Estrada do Lumiar, Campus do Lumiar, Edifício R, 1649‐038 Lisbon, Portugal
2Hovione FarmaCiencia SA, R&D Analytical Development, Estrada do Lumiar, Campus do Lumiar, Edifício R, 1649‐038 Lisbon, Portugal
3CICECO – Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810‐193 Aveiro, Portugal
4LAQV‐REQUIMTE, Department of Chemistry, NOVA School of Science and Technology, NOVA University Lisbon, 2829‐516 Caparica, Portugal
Summary: The present study successfully achieved the development and formulation of messenger RNA (mRNA) encapsulated in lipid nanoparticles (LNPs) for lung delivery through nebulization. tRNA‐LNPs and mRNA‐LNPs were produced via microfluidics and were dialyzed prior to nebulization. The formulation screening stage used transfer RNA (tRNA) as a model molecule and different formulations containing sodium chloride (NaCl), phosphate buffered saline (PBS), Poloxamer 188, Tween 80, sucrose, and/or arginine were evaluated. Among the tested conditions, the formulation with PBS and Tween 80 demonstrated the maintenance of tRNA‐LNPs colloidal stability upon nebulization when a dilution ratio of 1:10 was applied. The previous formulation was used for mRNA‐LNPs production and it was reported no loss of mRNA integrity throughout the product's lifecycle, effectively enduring mechanical stress during production, dialysis, and nebulization. The nebulized aerosol droplets, generated using a vibrating mesh nebulizer, displayed appropriate aerodynamic properties, with particle sizes ranging from 1 to 5 μm, thus ensuring effective lung deposition. Moreover, the nebulization of the formulation resulted in a Fine Particle Fraction of Emitted Dose (FPFED) of 51% and an Emitted Dose (ED) of 92% of the encapsulated mRNA.
Amorphous Spray Dried Microparticles for Nasal Delivery: Tackling Solubility Challenges while Targeting Systemic Nasal Absorption
Patricia Henriques1,2, Cláudia Costa1, António Serôdio1, Ana Fortuna2,3, Slavomíra Doktorovová1
1Hovione FarmaCiência SA, Lumiar, 1649‐038, Portugal 2Faculty of Pharmacy, University of Coimbra, 3000‐548 Coimbra, Portugal 3Coimbra Institute for Biomedical Imaging and Translational Research (CIBIT), Institute of Nuclear Sciences Applied to Health (ICNAS), University of Coimbra, 3000‐548 Coimbra, Portugal
Summary: Dry powder formulations for nasal drug delivery can be manufactured by spray drying, a technology that allows particle size control and the generation of amorphous solid dispersions for drug solubility enhancement. Targeting the turbinate area of the nasal cavity is relevant for drugs intended for systemic delivery that benefit from the fast absorption and quick onset of action provided by the vascularised nasal mucosa. Despite the current knowledge on spray dried powders for nasal delivery, the impact of different polymers on drug release performance is still not clear, as well as the deposition profile of this formulation strategy. Herein, amorphous spray dried microparticles were evaluated regarding their supersaturation behaviour and drug release performance, comparing with the corresponding physical blends. The lead formulation was additionally assessed regarding nasal deposition profile using the Alberta Idealized Nasal Inlet, while evaluating the impact of the angle of actuation from horizontal plane and inhalation flow rate. The results showed enhanced drug release performance for the amorphous formulations, where the polymer that sustained supersaturation for a longer period of time resulted in higher drug release. Nasal deposition studies with an angle of actuation of 45° and inhalation flow rate of 15 L/min showed high mass balance of 97% and high turbinate deposition of 55%. Overall, the results support that spray drying is an adequate strategy to manufacture microparticles with high turbinate deposition. Moreover, the combined performance evaluation strategy using dissolution, drug release and nasal deposition methodologies was adequate to characterise and select lead formulation candidates.
Developing a Combined Calu‐3 Barrier and Smooth Muscle Model of the Airways Utilising a Perfused Microphysiological System
Adeel A. Ahmed1, Emily L. Richardson2, Tomasz Kostrzewski2 & Darragh Murnane1
1University of Hertfordshire, Hatfield, AL10 9AB, UK
2CN Bio Innovations Limited, Cambridge, CB4 0WN, UK
Summary Successfully incorporating medium perfusion into the standard insert‐based air‐liquid (ALI) Calu‐3 model has resulted in a more physiologically representative pre‐clinical In‐Vitro model. The perfusion led to a tight epithelial monolayer working as an effective barrier to paracellular permeability and was maintained for up to 28 days. This tight barrier was established 7 days quicker than the static equivalent. qPCR analysis showed an increase in ZO1 and Claudin‐1 expression in the perfused model between days 7 and day 14, showing barrier formation was still occurring between the two timepoints. Using immunofluorescent staining, ZO1 was also shown to be significantly increased in expression in the perfused model at day 7 compared to the static model. Significantly, at day 7 the expression of the OCT1 cation transporter was greater in the perfused model than in the static model. Concurrently, airway smooth muscle cells were cultured in the basolateral face of an insert membrane to examine the feasibility of a micro‐physiological airways tissue co‐culture model. Cell population expansion and functionality was demonstrated using alpha‐smooth muscle actin staining with microscopy revealing the formation of a differentiated smooth muscle architecture. Culturing Calu‐3 under perfused conditions resulted in faster formation of a permeability barrier than static. Furthermore, bronchial smooth muscle cells were successfully cultured on both the underside of the insert and the base of the well plate. Together, the construction of these models will allow more advanced and predictive models suitable for bioavailability studies and disease modelling.
A lung‐to‐heart nanomedicine for chronic heart failure
Eride Quarta1,2, Paolo Colombo1,2, Ruggero Bettini1, Daniele Catalucci3,4, Claudio De Luca5, Paul Steendijk6, Barry Borlaug7, Fabio Sonvico1, Alessio Alogna8,9 & Francesca Buttini1
1Food and Drug Department, University of Parma, 43124 Parma, Italy;2PlumeStars s.r.l., Parma, 43125, Italy
3Institute of Genetic and Biomedical Research (IRGB), National Research Council of Italy,Milan Unit, Milan, 20138, Italy; 4IRCCS Humanitas Research Hospital, Milano, 20089, Italy;
5NanoPhoria S.r.l, Milano, 20123, Italy;
6Department of Cardiology, Leiden University Medical Center, Leiden, 2311G The Netherlands;
7The Department of Cardiovascular Medicine, Mayo Clinic, Rochester, MN 55906, USA;
8Herzzentrum der Charité, Department of Cardiology, Angiology and Intensive Care Medicine, Campus Virchow‐Klinikum, Augustenburgerplatz 1, 13353 Berlin, Germany; 93DZHK (German Centre for Cardiovascular Research), partner site; Berlin, 10785, Germany;
Summary: Peptides are highly selective molecules with the potential for a direct impact at the cardiomyocyte level. Yet, a strategy for effective delivery of therapeutic peptides to the heart is lacking. The aim of the present study was to assess tolerability and efficacy of an inhalable lung‐to‐heart nano‐in‐micro technology (LungToHeartNIM) for cardiac‐specific targeting of a mimetic peptide (MP), first in class for modulating impaired L‐type calcium channel (LTCC) trafficking, in a clinically relevant porcine model of heart failure (HF). HF with reduced ejection fraction (HFrEF) was induced in Göttingen minipigs by tachypacing over 6 weeks. In a setting of overt HFrEF (left ventricular ejection fraction (LV EF) 30 ± 8%), animals were randomized, and treatment started after 4 weeks of tachypacing. HFrEF animals inhaled either a dry powder composed of mannitol‐based microparticles embedding biocompatible MP‐loaded calcium phosphate nanoparticles (dpCaP‐MP) with high fraction of extra‐fine particles (<2μm) of 47.2 ± 0.7% or dpCaP without MP. Efficacy was evaluated by serial echocardiography and invasive hemodynamics. DpCaP‐MP inhalation restored systolic function, as shown by an absolute LV EF increase over the treatment time of 17 ± 6%, while reversing cardiac remodelling and reducing pulmonary congestion. The effect was recapitulated ex vivo in cardiac myofibrils from treated HF animals. The treatment was well tolerated, and no adverse events occurred. The overall tolerability of LungToHeartNIM along with the beneficial effects of the LTCC‐modulator point towards a game‐changing treatment for HFrEF patients, while demonstrating the effective delivery of a therapeutic peptide to the diseased heart.
Impact of protein aggregation on the immunogenicity of a human monoclonal antibody following pulmonary administration in mice
Sohaib Mahri1, Céline Cassiers1, Gracin Sandra2, Francine Uwambayinema5, François Huaux5, Mariam Ibrahim3, Sian Piper4, Antonio Llinas2, Markus Fridén2,6, Rita Vanbever1
1Université catholique de Louvain (UCLouvain), Louvain Drug Research Institute (LDRI), Advanced Drug Delivery & Biomaterials (ADBB), Brussels 1200, Belgium.
2AstraZeneca; Pharmaceutical Technology & Development; Inhalation Product Development; Gothenburg, Sweden
3AstraZeneca; R&D Biopharmaceuticals; Biopharmaceuticals Development; Gaithersburg, US
4AstraZeneca; R&D Biopharmaceuticals; Bioscience Asthma, Early R&I; Cambridge, UK
5UCLouvain, Institut de Recherche Expérimentale et Clinique, Louvain Centre for Toxicology and Applied Pharmacology, Brussels, Belgium
6Uppsala University, Department of Pharmacy, Uppsala, Sweden
Summary: Spray drying is commonly used to produce dry powder formulations for inhalation. However, this process subjects proteins to stresses likely resulting in the formation of protein aggregates. These aggregates can cause a loss of activity and an increased risk of immunogenicity. The impact of aggregates on the immunogenicity of therapeutic proteins delivered via the pulmonary route is not well understood. Therefore, our study aimed to specifically assess the effect of different levels of aggregates on the immunogenicity of the human monoclonal antibody NIP228 after repeated intratracheal administration in mice. NIP228 was labelled with NHS‐rhodamine and then aggregate‐free rhod‐NIP228 solution (No_agg) was spray‐dried both with and without 1% polysorbate 80 surfactant, to generate powders with high and low levels of NIP228 aggregates (referred to as Hi_agg and Lo_agg, respectively). Following repeated intratracheal administration of the No_agg, Lo_agg, or Hi_agg in mice, anti‐NIP228 antibody (ADA) levels in both sera and bronchoalveolar lavage fluids (BAL) were measured. The incorporation of PS80 significantly reduced the aggregate formation associated with spray drying and increased FPF <5 μm (78% for Hi_agg vs 87% for Lo_agg ). Interestingly, the level of aggregates did not significantly impact the magnitude of the anti‐drug antibody response in sera or BAL and the pulmonary route induced a stronger immune response compared to subcutaneous administration.
1Department of Pharmaceutical Sciences, University of Vienna, Josef‐Holaubek‐Platz 2, 1090 Vienna, Austria
2Bundesanstalt für Materialforschung und ‐prüfung / BAM Unter den Eichen 87 12205 Berlin, Germany
3Centre for Microbiology and Environmental Science Systems, University of Vienna, Josef‐Holaubek‐Platz 2, 1090 Vienna, Austria
Summary: Due to the difficulty of quantifying plastics in the environment, there is currently a lack of data on inhalation exposure to micro‐ and nanoplastics in indoor and outdoor aerosols. New and creative methodologies are required to address this data gap. Over the past year, we have taken our glass twin stage impinger on a grand tour of the continent, collecting aerosol samples from different locations off the Croatian coastline, in the Austrian countryside as well as in indoor climbing halls. It was hypothesized that certain exposure environments, such as inhalation of sea spray aerosol on the coast or dust from indoor climbing halls, will be hot spots of elevated exposure to rubbers, plastics and their additives. The development of a methodology for the quantification of respirable plastics and additives based on liquid impingement combined with granular fluid bed filtration is critically discussed. Preliminary results from our indoor sampling campaign revealed that the respirable fraction of air in climbing halls contained elevated concentrations particulates as well as additives derived from rubber shoes soles, as compared to control samples.
Climate change, air pollution, and their well‐documented harmful effects on human (lung) health
Zorana Jovanovic Andersen1
1Department of Public Health, University of Copenhagen, Øster Farimagsgade 5,1353 Copenhagen, Denmark.
Summary: Air pollution is the leading environmental risk factor for lung disease [1]. Long‐term exposure to air pollution from birth and throughout lifetime, cases inflammation and oxidative stress in the lung and suppresses immune system, slowing the development of lung function in children, and accelerating lung function decline in adults, and increases risk of a range of respiratory diseases including asthma, chronic obstructive respiratory disease (COPD), lung cancer, and acute lower respiratory infections. Short‐term exposure to air pollution, over several hours or days, can trigger symptoms in chronic respiratory patients including cough, wheezing, breathlessness, that may require need for medication, doctor care, hospitalization and even result in death. The World Health Organization (WHO) has warned that climate change is the biggest global threat to humanity in the 21st century [2]. Climate change is a major threat to lung patients, causing global warming and increasing frequency and duration of heatwaves, more frequent and extreme weather events, including wind‐ and dust storms, droughts, wildfires and related air pollution episodes, severe rainstorms and flooding, as well as introduction of new viruses and aeroallergens to new areas and prolonged aeroallergen seasons. All of these will have significant impacts on lung patients, alone or in combination with air pollution exposure. Continued education of respiratory health professionals on air pollution and climate change related impacts on lung health is crucial in helping them mitigate harmful impacts on their patients.
Do In‐vitro dissolution rates differ between beclomethasone dipropionate particles generated from solution‐based pMDIs formulated with HFA134a, HFA152a and HFO1234ze(E) propellants?
Hui Xin Ong1,2, Nirmal Marasini1, Lingzhe Rao3, Daniel Duke3, Damon Honnery3, Stephen W. Stein4, Benjamin Myatt5, Phil Cocks5 & Paul Young1,6
1Respiratory Technology, Woolcock Institute of Medical Research, Glebe, Sydney, NSW 2037, Australia
2Macquarie Medical School, Faculty of Medicine, Healthy and Human Sciences, Macquarie University, Sydney, NSW 2109, Australia
3Laboratory for Turbulence Research in Aerospace & Combustion (LTRAC), Department of Mechanical & Aerospace Engineering, Monash University, Melbourne, Australia
4Kindeva Drug delivery, 11200 Hudson Road, Woodbury, MN 55129
5Kindeva Drug Delivery, Charnwood Campus, 10 Bakewell Road, Loughborough, United Kingdom, LE11 5RB
6Department of Marketing, Macquarie Business school, Macquarie University, Sydney, NSW 2109, Australia
Summary: The switch from Hydrofluoroalkane 134a (HFA134a), with relatively high global warming potential (GWP), traditionally used in pressurised metered‐dose inhalers (pMDI) to low GWP propellants HFO1234ze(E) and HFA152a is currently underway, posing numerous scientific questions and challenges to formulation development. This study aims to investigate the morphological properties of maturated beclomethasone dipropionate (BDP) particles generated from solution‐based pMDIs formulated with three different propellants and the impact of these characteristics on in‐vitro drug dissolution. To achieve this, a modified Andersen Cascade Impactor (ACI) that incorporates SnapwellTM inserts on a 3D‐printed ACI plate was used to deposit and study the aerosol particles of specific aerodynamic size ranges produced from the three different pMDI formulations. The Snapwell insert was then used as a micro‐dissolution apparatus. The dissolution rate of the drug particles over four hours showed the following rank order: HFA134a > HFO1234ze(E) > HFA152a. This could be attributed to the morphologies of the drug particles found on stage 5 of the ACI where HFA134a produced maturated particles with the smallest geometric size of both the corrugated/rough (1.45 μm) and smooth/spherical particles (0.78 μm) with a higher percentage (53%) of rough/irregular particles in comparison with HFO1234ze(E) and HFA152a formulations. From these initial findings, a thorough investigation into the morphology and dissolution performance in combination with detailed physicochemical profiles and aerosol performance of the particles is required to understand the effect of the new propellants when formulating next the generation low GWP pMDIs.
Lactic Acid Bacteria Dry Powder Inhaler as Promising Tool for Pseudomonas aeruginosa Growth Inhibition in Pulmonary Dysbiosis
Stefania Glieca1, Eride Quarta1, Benedetta Bottari1, Elena Bancalari1, Erika Scaltriti2, Martina Tambassi2, Simona Bertoni1, Lisa Flammini1, Fabio Sonvico1 & Francesca Buttini1
1Food and Drug Department, University of Parma, Parco Area delle Scienze 27/a, Parma, 43125, Italy
2Risk Analysis and Genomic Epidemiology Unit, Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia‐Romagna, Strada dei Mercati, 13/A, Parma, 43126, Italy
Summary: The aim of the project was the development and characterization of inhalation powders containing a probiotic (Lactiplantibacillus plantarum, Lacticaseibacillus rhamnosus or Lactobacillus acidophilus) for the interaction with the lung microbiota and for the containment of P. aeruginosa (PA) growth in patients with cystic fibrosis. By optimizing a spray drying process applying a design of experiment, it was possible to maintain the viability of the probiotics, which decreased by only one order of magnitude after the process in comparison with the bacterial viability in the stock suspension.
Probiotic powders showed a high respirability (FPF approximately 50%) suitable for lung deposition. Interestingly, Lpb. plantarum powder showed a bactericidal activity on the three different PA strains while the other two probiotic powder were bacteriostatic. The effect could be explained by the fact that the Lpb. plantarum is less impacted by the spray drying process, and for this reason it has a greater ability to adapt to the culture medium, to grow and acidify it up to a pH of 3.7.
The work represents a promising starting point for considering a probiotic dry powder inhaler as a tool to contain the growth of pathogenic lung microflora. This approach could be beneficial during the suspension of inhaled antibiotic therapy in the treatment regimen of cystic fibrosis.
Repurposing bacterial lysates: an inhalation approach for respiratory infection management
Joana Pinto‐da‐Silva1, Melibea Berzosa2, Alberto Delgado‐López3, Carlos Gamazo3 & Ana Grenha1,4
1Centre for Marine Sciences, Universidade do Algarve, Campus Gambelas, Faro, 8005‐139, Portugal
2Wellcome Wolfson Institute for Experimental Medicine, Queen's University Belfast, Belfast, BT9 5AL, United Kingdom;
3Department of Microbiology, University of Navarra, Pamplona, 31008, Spain
4Faculdade de Ciências e Tecnologia, Universidade do Algarve, Campus Gambelas, Faro, 8005‐139, Portugal
Summary: Bacterial lysates (BL) are commonly used as immunomodulators in respiratory tract infection prevention. However, there is conflicting evidence for clinical efficacy, with the typical practice of oral administration limiting the potential to generate adequate lung protection. To improve the prospect of BL delivery inducing a beneficial immune response, a novel approach involving inhalation is proposed. Spray‐dried locust bean gum (LBG) microparticles (MP) were used to encapsulate BL and confer suitable properties for lung delivery. Furthermore, as a galactomannan, LBG will facilitate targeting of antigen presenting cells in the lungs through interaction with mannose surface receptors. The preservation of BL antigenicity was confirmed by direct agglutination using specific antibodies. A release assay was performed upon dry powder which was aerosolized onto a membrane in contact with a dissolution medium to represent lung conditions. A sustained release was observed with 80% release of BL after 6 h, reaching 85% release over 24 h. These data encourage the approach of repurposing BL for inhalation, with the aim of preventing respiratory infections.
Mucoadhesion and cytotoxicity of powders for nasal drug delivery
Alison B. Lansley, Xiaoqian Ding, Oluwatosin S. Adelowo & Matthew Ingram
Biomaterials and Drug Delivery Research and Enterprise Group, University of Brighton, Lewes Road, Brighton, BN2 4GJ, UK
Summary: The mucoadhesiveness and cytotoxicity of a range of powders was studied to test the hypothesis that mucoadhesive powders can be used safely in the nasal cavity to aid in the retention of the nasal formulations and improve the effectiveness of nasal products.
The mucoadhesiveness of a range of powders (hydroxypropyl methyl cellulose (HPMC 400 and 4000), pectin, carboxymethylcellulose, sodium salt (CMC) and colloidal microcrystalline cellulose (cMCC)) was measured using a Texture Analyser and their toxicity to a human bronchial epithelial cell line (16HBE14o‐) was measured using an MTT assay.
The mucoadhesiveness of the powders studied tended to decrease in the following order with HPMC 400 > HPMC 4000 > CMC > Pectin > cMCC > no sample. Generally, HPMC 400, HPMC 4000 and CMC were more mucoadhesive than cMCC (P < 0.05). The cytotoxicity of the powders appeared to depend on the volume of cell culture medium available to hydrate the powder. Apart from cMCC, which was not cytotoxic in the presence of 0, 50 and 100 μL medium, all powders were toxic in the absence of medium (p < 0.05). All powders, apart from pectin, were non‐toxic in the presence of 50 and 100 μL medium.
The mucoadhesiveness and cytotoxicity of different polymers, when applied in powdered form, is variable. Therefore, the choice of powdered polymer is important in optimising the retention of powdered nasal formulations while avoiding toxicity to the nasal epithelium.
Bacteria on the flight: local and international destinations?
Thomas Gutsmann1,2, Khanneh Wadinga Fomba3 & Christian Nehls1
1Research Center Borstel – Leibniz Lung Center, Division of Biophysics, Borstel, Germany
2Centre for Structural Systems Biology, Hamburg, Germany
3Leibniz Institute for Tropospheric Research, Department Chemistry of the Atmosphere, Leipzig, Germany
Summary: Dust storms can transport mineral dust (MD) from desert soil through the atmosphere to regions far from the dust origin. This impacts not only the quality of life, but also the health of millions of people in the “dust belt” and beyond: 330,000 deaths worldwide were associated with MD exposure in 2010 [1]. The climate change promotes a desertification and thereby an increase in global MD exposure. Respiratory, cardiovascular and allergic diseases can be observed in affected individuals. However, non‐pathogenic and pathogenic microbes are also transported with the dust [2]. These not only spread to new regions but may also lead to direct infections.
The “Dust‐Risk” consortium takes a highly interdisciplinary approach with direct public health relevance and aims to link the composition and properties of MD and associated microbes with its health risk. MD and health data have been collected in the Cape Verde Islands, which serve as a model region.
In our project part, we use atomic force microscopy and scanning electron microscopy to visualize and characterize the collected MD. With force spectroscopy, we determine the strength and type of molecular bonds between MD and different bacterial species that have been identified on the MD. Samples from different sampling sites are compared to distinguish between local dust and desert dust, and between microbes of local, marine, and desert dust origin.
Biophysical experiments help to understand how bacteria bind to MD and wrap in it to travel long distances through the atmosphere. This knowledge will be complemented by physicochemical, microbiological, toxicological, modeling, clinical and epidemiological data from further project parts to generate a composition‐based dust‐health‐risk‐index. The Cape Verde weather service will use this risk index to provide precise and specific advice and warnings to the population.
Evaluation of the anticancer activity of solid dispersions of chloramphenicol derivative with commonly used inhaled excipients
Arwa Al Khatib1,2, Mohamed El Tanani2, Muhammad Yaqoob3, Hisham Al‐Obaidi1
1Department of Pharmacy; School of Chemistry, Food and Pharmacy, University of Reading
2Pharmacological and Diagnostic Research Centre, Faculty of Pharmacy, Al‐Ahliyya Amman University, Amman 19328, Jordan
3Interaction Chem Pharm Ltd, Reading RG2 0QX, UK
Summary: The purpose of this study is to investigate the anti‐lung cancer effect of chloramphenicol derivatives such as N‐phenyl 2,2 dichloroacetamide (PDA). A design of experiment (DoE) approach was used to form amorphous solid dispersions with mass ratios of three commonly used excipients (l‐leucine, trehalose, and mannitol) ranging from 30% to 5% using three fluid nozzle spray drying. The effects of each excipient on IC50 in A549 cell lines were assessed. The anti‐proliferative assays of the free PDA showed a concentration‐dependent trend. Physicochemical analysis using EDX‐SEM analysis, confirmed that the drug was deposited uniformly on the surface. However, it was shown that the spray‐dried particles varied in their surface characteristics depending on the different ratios of drug and excipients used. Bulk uniformity was confirmed using Raman mapping in which it showed that the drug was uniformly distributed among the spray‐dried particles. In situ particle analysis using focused beam reflectance measurement (FBRM) showed that these particles had biphasic dissolution, indicating the potential to cause a fast release of the drug followed by a slower release.
Effects of cyclodextrins and human serum albumin on solubilising and aerosolising cannabidiol
Waiting Tai1, Jonathon Carl Arnold2, Hak‐Kim Chan1 & Philip Chi Lip Kwok1
1Advanced Drug Delivery Group, Sydney Pharmacy School, Faculty of Medicine and Health, The University of Sydney, NSW 2006, Australia
2Lambert Initiative for Cannabinoid Therapeutics, Brain and Mind Centre, The University of Sydney, NSW 2050, Australia 3Discipline of Pharmacology, Sydney Pharmacy School, Faculty of Medicine and Health, The University of Sydney, NSW 2006, Australia
Summary: Inhaled cannabidiol (CBD) has attracted much interest due to its promising therapeutic potential. The major formulation challenge is its low aqueous solubility, so solubilisers are required. In this study, the effects of three cyclodextrins (881 μM β‐cyclodextrin, methyl‐β‐cyclodextrin, and 2‐hydroxypropyl‐β‐cyclodextrin) and 15 μM human serum albumin on CBD solubility were evaluated by the shake‐flask method. The highest CBD saturated concentration was achieved by methyl β‐cyclodextrin at 14.6 ± 0.2 μg/mL, followed by human serum albumin at 9.7 ± 0.1 μg/mL. Methyl‐β‐cyclodextrin and human serum albumin were then spray freeze dried with CBD to obtain amorphous Formulations M and H, respectively. The aerosol performance of the two formulations was determined using the Next Generation Impactor. Although the mass median aerodynamic diameter of Formulation M was significantly smaller than that of Formulation H (3.7 ± 0.1 μm vs 4.2 ± 0.3 μm), but both had comparable geometric standard deviations (2.3 ± 0.0 vs 2.4 ± 0.1). With about 90% CBD emitted from both formulations, the fine particle fraction <5 μm of Formulation M was significantly higher than that of Formulation H (53.5 ± 1.5% vs 30.2 ± 1.00%). Since methyl‐β‐cyclodextrin achieved higher CBD solubility and better powder dispersion performance, it may be used to deliver other cannabinoids.
In Vivo Biodistribution of Inhaled Colistin‐Endolysin Combination
Pengfei Zhang1, Ping Zeng1 & Sharon S. Y. Leung1
1School of Pharmacy, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong SAR
Summary: The rapid emergence of multidrug‐resistant (MDR) bacteria, particularly the gram‐negative (G‐ve) ones, has significantly increased the medical challenges in managing their associated pneumonia. Combination therapy of colistin with bacteriophage‐derived endolysins (lysins) has recently attracted significant attention to combat infections caused by MDR G‐ve bacteria. To maximize the synergy of the combination treatment, it is essential to maintain the two agents at an optimal mixing ratio at the infection sites. In the present study, we investigated the lung retention time of colistin and LysAB2 lysin in healthy BALB/c mice via intratracheal (IT) aerosolization and intravenous (IV) injection. Anaesthetized mice were dosed with fluorescence‐labelled colistin (with sulfo‐cy5) and LysAB2 (with sulfo‐cy7). At designated time points (1, 2, 4, 8, and 24 h), the distribution of both agents was monitored through fluorescence measurement with an in‐vivo animal imaging system (IVIS). Our results showed colistin and LysAB2 shared similar biodistribution profiles regardless of the administration routes. Compared with IV injection, colistin and LysAB2 delivered via IT aerosolization had apparently higher and longer lung exposure. The promising data suggest that the optimal mixing ratio of the two agents from in‐vitro setting can be applied for in‐vivo experiments with minimum adjustment.
Inhaled Lacticaseibacillus rhamnosus‐derived postbiotics as novel strategy to tackle Pseudomonas aeruginosa lung infection in cystic fibrosis
1Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, Via S. Zeno 35‐39, Pisa, 56123, Italy
2Department of Pharmacy, University of Pisa, Via Bonanno 33, Pisa, 56126, Italy
Summary: Probiotics or their metabolic products, also referred to as postbiotics, have recently gained interest as potential antimicrobials, given the rise of multidrug‐resistant pathogens and the loss of efficacy of many antibiotics in the clinic. This study aimed to investigate the antimicrobial activity of cell‐free supernatants from Lacticaseibacillus rhamnosus (CFS‐LR) against Pseudomonas aeruginosa strains isolated from the sputum of cystic fibrosis (CF) patients. In addition, we aimed at developing a liquid CFS formulation for aerosol administration, in view of future implementation of CFS as inhaled therapy to treat or prevent P. aeruginosa lung‐infections. When tested in an in‐vitro P. aeruginosa lung infection model, CFS‐LR exhibited a marked antibacterial effect reducing P. aeruginosa viable count of approximately 2 Logs within 7h of incubation at the dilution 1:8, while causing complete eradication of P. aeruginosa at the dilution of 1:4. Cytotoxicity assessment of CFS‐LR against monolayers of NCI‐H441 human distal lung epithelial cells demonstrated that the dilution of 1:8 was well tolerated, while marked levels of cell‐death were observed at the 1:4 dilution. The aerodynamic distribution of nebulized CFS‐LR was evaluated by a twin‐stage impinger, using an Aerogen® mesh nebulizer. As much as 76% of the nebulized CFS‐LR solution was deposited in the second stage of the model suggesting a satisfactory distribution in the profound airways. Though CFS‐LR lyophilization caused partial reduction of the bactericidal potential, marked levels of antimicrobial activity were retained. In conclusion, these findings support the study of CFS aerosol delivery as an innovative strategy to control P. aeruginosa lung‐infections.
Pulmonary delivery of Lactobacillus rhamnosus GG with vibrating mesh and jet nebulisers
Alex Seungyeon Byun1, Luis Vitetta1, Hak‐Kim Chan1 & Philip Chi Lip Kwok1
1School of Pharmacy, Faculty of Medicine and Health, The University of Sydney, New South Wales 2006, Australia
Summary: Probiotics are commonly used to improve gastrointestinal symptoms and maintain health. Studies have frequently demonstrated the positive effects that oral probiotics exert not only in the gastrointestinal tract but also the respiratory tract. There is sufficient evidence to support that oral probiotics can reduce the incidence and severity of upper respiratory tract infections, thereby reducing antimicrobial use. In the past decade, some animal studies have examined the effects of intranasal probiotics on the respiratory tract, but nebulised probiotics have not yet to be explored. Therefore, this study compared the dose output and aerosol performance of probiotics nebulised from vibrating mesh and jet nebulisers. The vibrating mesh nebuliser (VMN) consistently produced a higher output than the jet nebuliser (JN) for Lactobacillus rhamnosus GG in both phosphate‐buffered saline (PBS) and normal saline (0.9% w/v sodium chloride in water) at 23.18 ± 0.99% and 10.43 ± 3.99%, respectively. For both nebulisers, probiotic in PBS outperformed its saline counterpart. A similar trend was observed with the fine particle fraction as it was higher for PBS (12.95 ± 0.96% for VMN; 11.21 ± 0.18% for JN) than normal saline (7.51 ± 2.01% for VMN; 7.03 ± 2.48% for JN) for both nebulisers, suggesting that aerosol performance of nebulised probiotics may also depend on the medium in which the probiotics were suspended in. Therefore, this study demonstrated that the type of nebuliser as well as the vehicle of the probiotic suspension affected the effectiveness of nebulisation.
Novel Inhalable Formulations for Targeted Delivery of Antibiotics to the Lung: A Response to the Global Tuberculosis Commitment
Sharareh Salar‐Behazdi1,2, Carolina Corzo1, Eleonore Fröhlich1,3, Dirk Lochmann4, Sebastian Reyer4, Andreas Zimmer2
1Research Center Pharmaceutical Engineering GmbH, Inffeldgasse 13, 8010, Graz, Austria
2Institute of Pharmaceutical Sciences, Department of Pharmaceutical Technology and Biopharmaceutics, University of Graz, Universitätsplatz 3, 8010, Graz, Austria
3Center for Medical Research, Medical University of Graz, Stiftingtalstraße 24, 8010, Graz, Austria
Summary: This work is a response to the significant increasing demand on the inhalable antibiotic drugs for efficient localized delivery to the lung. We aimed to engineer safe and novel lipid‐microparticles loaded with rifampicin via spray‐drying, to be administered as a carrier‐free dry powder for inhalation. Despite the well‐known strong technical limitations of spray‐drying of lipids, a completely lipid‐based formulation was successfully developed, using diglycerol full ester of behenic acid and glyceryl stearate citrate as emulsifier.
Exceptional thermal and solid‐state properties of lipid as a homogeneous system with high melting temperature, low super‐cooling (ΔT = Tm‐Tc) and absence of polymorphism were the key factors to avoid softening of material during spray drying and to gain a high yield of 83%. The achieved mass median aerodynamic diameter and median geometric size of particles were 2.36 μm and 2.05 μm, respectively. Those properties together with the negative surface (‐50.03 mV) of particles were defined as critical factors for deposition in the deep lung and targeted delivery of rifampicin to alveolar macrophages. The fine particle fraction was 79.5%. The in vitro uptake of particles into THP‐1 macrophages was confirmed by fluorescence imaging and by determination of the rifampicin content inside the alveolar macrophages. The in vitro safety of particles was confirmed based on cytotoxicity studies on alveolar and bronchial epithelial cells (A549 and Calu‐3 cell lines, respectively), as well as on alveolar macrophages (THP‐1 cells).
This study paves the way to extend the use of lipid‐based formulations for advanced targeted delivery to the lung.
Design of in vitro lung mucus‐microbiota model to assess efficacy of inhaled antibiotics
Mahmoud H. Abu Elella1, Simon Andrews2, Glyn Barrett2, Vitaliy Khutoryanskiy1, Hisham Al‐Obaidi1
1School of Pharmacy, University of Reading, Reading, RG6 6UR, UK
2School of Biological Sciences, University of Reading, Reading, RG6 6UR, UK
Summary: Over a few decades, many scientific experiments worldwide consumed huge number of animals for the testing of various inhaled drugs. Among them, several studies require to scarify animals to extract mucosal tissues for examining the action of inhaled powders. Recently, designing alternative methods instead of animal models is an attractive strategy. For instance, some synthetic 3D polymeric hydrogels have similarities to the chemical structure of the mucus that lines‐mucosal tissues. Here, we aim to design mucus‐mimicking hydrogels, simulating lung‐microbiota layer, and adapted a Andersen cascade impactor (ACI) to assess the deposition of aerosolised ciprofloxacin powders. Ciprofloxacin powders were prepared using two and three fluid nozzle spray dryers. Moreover, via free‐radical polymerisation technique, hydrogels were synthesised by copolymerising N‐acryloyl‐glucosamine with 2‐hydroxyethylmethacrylate. In our model, we used E. coli, as a bacterial model, to simulate the lung's mucus‐microbiota layer and modified Gamble's solution with dipalmitoylphosphatidyl choline, and citrate as simulated lung fluid (SLF). The findings revealed successful preparation of highly porous mucosa‐mimetic hydrogels via the elucidation of the surface structure with Cryo‐SEM techniques. After mixing the hydrogels with SLF, the in vitro mucus‐microbiota‐ACI model was validated by administering lactose. The deposition results showed minimal differences in deposition between a control and modified ACI, and bacteria were completely eradicated for samples treated with ciprofloxacin, demonstrating the diffusion of deposited ciprofloxacin within porous hydrogel layers.
Advancing nasal formulation testing: What equipment to use for delivered dose assessment?
Niklas Baltz & Regina Scherließ
Department of Pharmaceutics and Biopharmaceutics, Kiel University, Gutenbergstraße 76, 24118 Kiel, Germany
Summary: Uniformity of delivered dose is crucial in nasal products. Regulatory guidelines require to demonstrate uniformity, but the choice of uniformity assessment method is left open. This study evaluated different equipment for determining the delivered dose of nasal formulations and aimed to identify a suitable setup for all types of formulations, as a step towards selecting standard universal method(s) for assessing nasal delivered dose. Three formulations, a multidose solution (Pollicrom), a multidose suspension (MometaHexal) and a single dose powder (Baqsimi), were tested. Setups including the Dosage Unit Sampling Apparatus (DUSA), glass expansion chamber, Nasal Spray Dose Collector (NSDC), custom‐made setups, and the Kiel Nasal Inlet (KNI) were evaluated. Measurements were performed with and without airflow, as applicable. Setups without airflow showed high dose recovery. The KNI exhibited high dose recovery regardless of airflow, while DUSA and the custom‐made frit setup showed limitations with leakage and formulation loss under airflow. While all setups operated without airflow were suitable for measuring dose uniformity of the nasal solution and suspension, testing with airflow could be important for nasal powders and pressurised metered dose sprays to retain the dose in the corpus of the equipment. Future studies will focus on multidose nasal powders and pressurised metered dose sprays to extend this work. This study provides insights into equipment selection for assessing delivered dose in multidose nasal sprays and single dose nasal powders.
Potential use of cannabinoids to combat against methicillin‐resistant Staphylococcus aureus pneumonia: A pre‐formulation study
Pancy Tsz Hei Kwong1, Theerthankar Das2, Jonathon Carl Arnold3, Hak‐Kim Chan1 & Philip Chi Lip Kwok1
1Advanced Drug Delivery Group, Sydney Pharmacy School, Faculty of Medicine and Health, The University of Sydney, NSW 2006, Australia
2Ingham Institute for Applied Medical Research, Microbiology and Infectious Diseases, School of Medicine, Western Sydney University, NSW 2170, Australia
3Lambert Initiative for Cannabinoid Therapeutics, Brain and Mind Centre, The University of Sydney, NSW 2050, Australia
Summary: Methicillin‐resistant Staphylococcus aureus (MRSA) is a major cause of hospital‐acquired lung infections. MRSA is likely to form a biofilm in the lungs, rendering the infection more difficult to eliminate which often results in treatment failure. Due to the challenges in treating MRSA pneumonia, the administration of high doses of antibiotics frequently leads to severe adverse effects. In this study, potentially more tolerable, non‐psychoactive cannabinoids – cannabidiol (CBD), cannabigerol (CBG), and cannabinol (CBN) were investigated for their activity against a MRSA isolate cultured in a biofilm. CBN was discovered to be the most potent candidate with anti‐MRSA biofilm activity, shown by reductions in biofilm biomass and bacterial viability. In particular, CBN exhibited efficacy in killing more than 80% of the bacteria in the biofilm at 1 μg/mL, whereas CBD and CBG were less effectively, with 10% and 60% reduction in viability, respectively. Only CBN demonstrated significant biofilm dispersal effect at all concentrations. With their penetration through the extracellular matrix and potent antibiofilm activity against MRSA, cannabinoids present an exciting opportunity for the development of inhalable formulations to treat bacterial pneumonia.
In Silico Modelling to Aid Product Development Strategies for Low Global Warming Potential Metered Dose Inhalers
Neha Patel1 & Alexander Richford1
1Kindeva Drug Delivery Ltd., Derby Road, Loughborough, Leicestershire, LE11 5SF, United Kingdom
Summary: The current expectation is that new metered dose inhalers (MDIs) containing existing drug substances re‐formulated with alternative low global warming potential (GWP) propellants (HFA‐152a and HFO‐1234ze) will have similar in‐vitro and/or pharmacokinetic (PK) performance to the already‐approved HFA‐134a and HFA‐227 reference products. In‐silico PK modelling provides a development tool for predicting which new product configurations are most likely to be comparable to existing products. Preludium™ has been used to calculate lung regional deposition from clinically relevant in‐vitro cascade data, which in turn has been used to develop an inhalation PK model for salbutamol products that can predict PK plasma profiles, differentiate product strengths and different inhalation dosage forms. An exponential error model has then been applied to the PK model to simulate expected between (BSV) and within subject variability (WSV).
In‐silico simulation has been used to compare various low GWP MDI test product configurations in terms of predicted PK performance. Using the exponential error model, it is possible to simulate variance that could be expected in a PK study and establish which test product configuration is most likely to achieve PK bioequivalence (BE) compared to a reference product. This may be particularly useful for the development of bioequivalent new low GWP MDI products.
A Novel Microdosing Device for Precise Dosing in Inhalation Therapy
A. Pietropaolo1,2, A. M. Healy2,3, C. Ehrhardt2, F. Sonvico1
1University of Parma, Department of Pharmacy, 43121 Parma, Italy
2School of Pharmacy and Pharmaceutical Sciences, Trinity College Dublin, Dublin 2, Ireland
3SSPC, The Science Foundation Ireland Research Centre for Pharmaceuticals, Ireland
Summary: This study aimed to evaluate the novel SAK‐1 microdosing device, which was designed to accurately dose small powder quantities, e.g. to fill capsules and other receptacles. Manual weighing methods commonly used in laboratories for microdosing often suffer from precision errors and are time‐consuming procedures. However, industrial scale instruments that are designed for this purpose can be impractical in a laboratory setting due to the large quantities of powders required and the high cost of tooling for additional dosing volumes, which is particularly true for drum dosing systems. In the current project, the SAK‐1 device was assessed for its accuracy and reproducibility in dosing four different grades of lactose powder into DPI capsules. Particle size analysis was carried out and powder flow characteristics were evaluated to provide insight into the critical powder parameters which could potentially affect the SAK‐1's performance. The results highlight differences in powder properties, including particle size distribution, with median particle size ranging from 11 μm (for InhaLac® 400) to 138 μm (for InhaLac 120). Flow characteristics also varied between different lactose samples. Basic Flowability Energy (BFE) ranged from 85 mJ (for InhaLac 400) to 559 mJ (for InhaLac 230). InhaLac 400 had the highest measured cohesion and compressibility and presented challenges during manual weighing due to its high cohesion, making it less flowable and more difficult to handle. Additionally, InhaLac 400 exhibited a tendency to form aggregates, leading to powder clumping. These issues were not encountered when using the SAK‐1 microdosing device. Capsules were filled to three targeted fill volumes for the four lactose grades. The relative standard deviations (RSDs) for the measured fill weights ranged from 0.5% to 2.6%. This indicates a high level of microdosing precision, despite variable particle size and flow characteristics of the dosed powders. The fill weight RSD may be influenced by a number of manual operator parameters and the chemical and physical characteristics of the powder. Due to the complexity of the factors affecting the filling process, it was not possible to establish a direct correlation between a single micromeritic parameter and the precision of the filling operation. In conclusion, the SAK‐1 microdosing device offers a promising solution for precise and efficient powder dosing in laboratory settings, particularly for dry powder inhalation formulations, with the added benefit that it can also simulate drum dosing systems, thus offering a seamless upgrade path as the project develops.
Development of a Direct in Vessel Aerosol Collection Apparatus for Dissolution Testing
Mårten Svensson1, Lena Heintz1 & Kjell Fransson2
1Emmace Consulting AB, Scheelevägen 22, 223 63 Lund, Sweden
2AB FIA, Scheelevägen 6, 223 63 Lund, Sweden
Summary: Dissolution testing of inhaled drugs has received significant increased interest with respect to regulatory science over the last ten years. However, a simple and standardised dissolution apparatus with discriminatory power is still not widely available to industry or academia. An aerosol collection apparatus was designed to capture directly into a liquid medium in a standard dissolution vessel, to avoid potential issues known when using a filter surface for collection. The new apparatus demonstrated good agreement with an existing paddle over disk dissolution method once normalised for collected dose, but further work will be needed to ensure more efficient total dose capture and therefore lower losses to the exhaust filter.
A device screening method for predicting delivery of inhaled mRNA via a nebuliser
Imco Sibum1, Bernhard Mullinger1 and Nicolas A. Buchmann1
1Resyca BV, Colosseum 23, Enschede, 7521 PV, the Netherlands
Summary: A device screening method is presented for initial comparison of full‐length mRNA lung delivery via nebulisation route. To this end, aerosol data was collected based on manufacturer information and subsequently used in deposition modelling. Several nebuliser systems with different aerosolisation technologies were included into the study. Aerosol data shows that the MMAD was slightly larger for the soft mist spray chip technology as compared to the vibrating mesh devices. However, deposition modelling results indicated that lung deposition is higher for the soft mist nebuliser. Using mRNA post nebulisation integrity data, reported for these three devices, predicted full‐length mRNA lung deposition could be calculated. This calculation shows that the Pulmospray™ results in 51.3% mRNA lung deposition compared to eFlow's 2.3% and Solo's 11.7 %, as percentage of fill dose. Deposition of full‐length mRNA is critical, as only full‐length mRNA results in the wanted biological‐active protein and efficient full‐length mRNA deposition depend on the choice of the delivery device.
The use of microfluidic method to prepare PEG12KL4/siRNA complexes for pulmonary delivery
Cheng MA1,2, Michael YT CHOW1,2 & Jenny KW LAM1,2
1UCL School of Pharmacy, University College London, 29‐39 Brunswick Square, London, WC1N 1AX, UK
2Department of Pharmacology and Pharmacy, LKS Faculty of Medicine, The University of Hong Kong, 21 Sassoon Road, Hong Kong SAR
Summary: Small interfering RNA (siRNA) has huge potential to treat pulmonary diseases, but the lack of a reproducible method to prepare the siRNA delivery system limits the application of siRNA therapy. Previous studies showed that PEG12KL4, a synthetic cationic peptide, is a promising vector for delivering RNA to the lung with excellent transfection efficiency. This study aimed to develop a reproducible and scalable method to prepare PEG12KL4/siRNA complexes by adopting a microfluidic approach for mixing the peptide and siRNA. The objectives were to (i) understand how the inlet flow rate ratio of peptide to siRNA and the total outlet flow rate affect the particle size distribution of the complexes; and (ii) compare the transfection efficiency of PEG12KL4/siRNA complexes prepared by microfluidic method and conventional pipette mixing method on human lung epithelial cells. The results showed that a low inlet flow rate ratio facilitated the formation of complexes with smaller particle sizes and better reproducibility, whereas the total outlet flow rate used in this study did not have any significant impact on the particle size. In addition, the transfection study of the PEG12KL4/siRNA complexes prepared with the microfluidic approach showed a similar gene silencing efficiency as those prepared by the pipette mixing method, suggesting that the microfluidic approach is a feasible method in producing PEG12KL4/siRNA complexes.
Impact of the Aptar Digital Health (ADH) respiratory platform on adult asthma healthcare cost
1Aptar Digital Health, 245 Main Street, 2nd Floor, Cambridge, Massachusetts 92142, USA
2Penn State University College of Medicine, 500 University Drive, Hershey, Pennsylvania 17033, USA
3Clyptek, Inc., 323 NE 34th Street # 912A, Fort Lauderdale, Florida 33308, USA
4Aptar Digital Health, 22 Quai Gallieni, 92150 Suresnes, France
5Aptar Pharma, 250 NY‐303, Congers, New York 10920, USA
Summary: Asthma is a common chronic respiratory disease impacting 334 million people globally (4% of the world's population), and resulting in over 400,000 deaths annually.1,2,3 In the United States (US), the total annual economic cost of asthma is $105 billion, including $65 billion in direct medical cost, $37 billion in asthma‐related mortality, and $4 billion in absenteeism cost (inflated to 2022 cost based on Bureau of Labor Statistics (BLS) medical care consumer price index (CPI)).4,5 Total annual health care costs (inpatient hospitalizations, emergency room visits, outpatient visits, and medications) for asthma patients with exacerbations is $14,359 and without exacerbations is $7,802 (inflated to 2022 values based on BLS medical care CPI).6,7 Across all asthma types and severities, 12.5% and 8.4% of asthmatics in the US and UK, respectively, experienced ≥1 exacerbations annually.8 Medication adherence is estimated at 50% worldwide.9 Improved controller medication adherence and Asthma Control Test (ACT) scores are correlated with decreased asthma exacerbations.10,11,12 Digital therapeutics (DTx) are utilized to improve controller medication adherence and ACT scores, which correspond to a potential reduction in exacerbations, and to decrease rescue medication usage, healthcare utilization, and societal and healthcare costs. A prospective, single cohort observational study evaluated 104 adult patients utilizing the Aptar Digital Health (ADH) respiratory platform, a DTx, for 90 days. Results demonstrated improved controller medication adherence (compared to global average), ACT scores, and decreased rescue medication usage. Improvements in controller medication adherence correlated with cost savings due to reduction in exacerbations.
A Novel Mathematical Model to Predict The Particle Size Distribution of pMDI Aerosols
H Chizari1, H.K. Versteeg2, W. Malalasekera2 & B. Gavtash1
1Kindeva Drug Delivery, Derby Road, Loughborough, LE11 5SF, UK
2Wolfson School of MEME, Loughborough University, Ashby Road, Loughborough, LE11 3TU, UK
Summary: This paper introduces a fundamental method to estimate droplet size distribution in aerosols from pressurised metered dose inhalers (pMDIs). Accurate droplet size representation is vital for estimating drug deposition. Using the instability equation by Reitz et al., we calculate droplet size based on fast‐growing wave modes. The slip velocity, a crucial parameter, is adjustable in our model. Comparing predictions with experimental data shows good agreement for HFA134a and HFA227ea aerosols with minor slip velocity adjustments. This model is essential for predicting droplet distribution and calculating drug deposition in the USP‐IP throat.
Assessment of Aerosol Characterisation and Delivery Performance of a Solution Using Different Configurations of a Breath‐Actuated Mesh Nebuliser
Edgar H. Cuevas Brun, Ciou‐Ting Wang, Yen‐Ting Chen, Chih‐Tsen Liu, Yu‐Hsuan Lin & Yuan‐Ming Hsu
HCmed Innovations Co. Ltd., Rm. B, 10F., No.319, Sec.2, Dunhua S. Rd., Taipei City, 10669, Taiwan
Summary: Delivery performance of a formulation using a mesh nebuliser is dependent on the physical chemical properties of the liquid substance. Several factors such a viscosity and surface tension can influence the performance of a delivery system affecting its effectiveness. Therefore, counting with a set of customisation approaches within a nebuliser platform can allow to explore and compute the outcome for parameters with different configurations. These customisation approaches could extend not only to hardware modifications, but also firmware changes within the algorithm of the nebuliser. The latter one being of particular significance for newer smart breath‐actuated mesh nebulisers that are equipped with novel functions. In this study, a commonly nebulised bronchodilator was used to explore the delivery performance of the AdheResp smart breath‐actuated mesh nebuliser platform, focusing on a two‐stage study to demonstrate the influence of customisation capabilities. The first stage consisted of a hardware adjustment, by testing a selection of mesh pore sizes and recording its impact on droplet size distribution. At the second stage, firmware modifications in triggering span during inhalation were also tested to collect data on the delivery performance and implications for each configuration that generated information about the correlation between triggering span and treatment time, and its later influence on delivery rate through delivered dose assessment. Consequently, the overall scope of the study presents the benefits of an exploratory work at the development stage of a new drug‐nebuliser combination product, aiming to identify the most suitable pair for the targeted delivery requirements.
Inhaler Design Considerations for Humidity Exposed Spray Dried Vaccine Powders
Varsha Singh1, Mikael Ekström2, Simon Karlsson2, Orest Lastow2, Myrna Dolovich3,4 & Michael Thompson1
1Department of Chemical Engineering, McMaster University, Hamilton, Ontario, L8S 4L7, Canada.
Summary: With the advancement of viral vaccines as thermally stable powders for improved stockpiling and distribution, a timely opportunity arose to evaluate the design of a commercially available dry powder inhaler, IcoOne (Iconovo AB) for their administration. In this study, the original design was compared to three modified designs for their performance in dispersing two well‐characterized dry powder vaccine formulations offering a prolonged period of bioactivity. The two formulations tested in this study, mannitol/dextran (MD) and leucine/lactose/trehalose (LLT), were chosen because of their novelty as next‐generation vaccines and differing cohesive natures in the presence of moisture. The inhaler designs were compared for dispersion of freshly spray dried placebo powders vs powders exposed to 50% relative humidity. The efficiency of the inhalers was analyzed based on aerosol performance using Next Generation Impactor (NGI), and uniformity of the emitted dose. The original (commercial) design was found to outperform the modified versions, dispersing both fresh and humidity affected powders efficiently.
Comparing The Aerosol Front Velocity Of Dry Powder Inhalers With The Corresponding Velocity Of Pressurised Metered Dose Inhalers And Soft Mist Inhalers
Summary: The core task of an inhaler is to generate a pharmaceutical aerosol. Observing the aerosol leaving the mouthpiece helps the engineer, the doctor and finally the patient. The latter benefits from optimized efforts by the healthcare professionals, providing better inhalers and detailed instructions as well as training. In the past, only aerosol plumes leaving active inhalers, e. g. pressurised metered dose inhalers (pMDIs) and soft mist inhalers (SMIs) have been investigated. This study adds the air flow which is generated by the patient during intended use. Dry powder inhalers (DPIs) were included. Experimental: For simplicity, rates of 30, 60, and 90 L/min of constant air flow were selected. Instead of sealing the mouthpieces by the patients' lips now the inhalers were enclosed and sealed to the enclosure where the lips are located. These enclosures were filled with air at the selected flow rate. Results: At 10 cm distance of the mouthpiece, the Respimat SMI had the slowest front velocity (0.99 m/s) and the longest spray duration (1447 ms) compared with pMDIs (velocity: 3.65–5.09 m/s; duration: 227–270 ms) and DPIs (velocity: 1.43–4.60 m/s; duration: 60–757 ms). With increasing flow rates, SMI aerosol velocity increased (maximum 2.63 m/s), the duration was not affected; pMDI aerosol velocity and duration were unaffected, and DPI aerosol velocity tended to increase, with a more variable impact on duration. Limitations: pushing air instead of sucking modifies the air density. This is of low relevance as the inhalers are operated at ambient pressure approximately.
Assessment of the Delivery Performance of a Novel Soft Mist Inhaler Using a Pre‐Filled Syringe Based Container Closure System
Pragathi Gurumurthy1, Knut Sommerer2, Imco Sibum1, Nicolas A. Buchmann1 & Bernhard Müllinger1
1Resyca BV, Colosseum 23, Enschede, 7521 PV, The Netherlands
2Actarmo Medical GmbH, Robert‐Koch‐Allee 29, Gauting, 82131, Germany
Summary: Novel drug developments (e.g. biologics) are primarily delivered as water‐based liquid formulations. Soft Mist Inhalers (SMIs) deliver the water based liquid formulations through a slow‐moving cloud of aerosol that is generated solely by mechanical energy. Resyca's novel soft mist inhaler with a pre‐filled syringe (PFSI) delivers high doses of liquid drugs that can be aseptically filled. This inhaler has high delivery efficiency leading to a high lung deposition. The particle size distribution measured by laser diffraction gives a VMD of 4.27 μm with a high FPF of 65.20% at 30 L/min. The average emitted dose measured gravimetrically at 30 L/min over multiple devices was 30.41 ± 0.24 mg. The regional respiratory deposition calculated from the obtained aerosol data shows a high lung deposition of 62.88%.
Influence of Fluid Physiochemical Properties on Aerosolization Performance of a Novel Soft Mist Inhaler
Varsha Komalla1, Imco Sibum3, Bernhard Muellinger3, Wietze Nijdam4, Vishal Chaugule5, Julio Soria5, Nicolas A. Buchmann3, Hui Xin Ong1,2, and D. Traini1,2
1Respiratory Technology, Woolcock Institute of Medical Research, Sydney, NSW 2037, Australia
2Macquarie Medical School, Department of Biological Sciences, Faculty of Medicine, Health and Human Sciences, Macquarie University, NSW 2109, Australia
3Resyca B.V., Enschede, the Netherlands
4Medspray B.V., Enschede, the Netherlands
5Laboratory for Turbulence Research in Aerospace and Combustion (LTRAC), Department of Mechanical and Aerospace Engineering, Monash University, Clayton Campus, Melbourne, VIC 3800, Australia
Summary: The purpose of this work is to provide a better understanding of the effects of different levels of excipients affecting fluid properties and inhalation flow rates on the aerosol performance of the novel Soft Mist Inhaler (SMI), PulmosprayTM. Water for injection (WFI) was used as a solvent and the excipients included were varying concentrations of ethanol (5 to 50% v/v), glycerol (5 to 30% w/v), sodium chloride (0.3 to 9% w/v) and span 20 (0.01 and 0.05% v/v). The excipients were evaluated to elucidate the impact of these additions to the solution formulation properties on inhalation aerosol variables (Particle Size Distribution, PSD, Dv10, Dv50 and Dv90) at two different flow rates (15 and 30 L/min). In general, an increase in the concentration of any excipients added increased osmolality. The addition of Span 20 and ethanol individually to WFI resulted in a decrease in surface tension, and the addition of glycerol increased viscosity. In summary, the characteristic of aerosol generation and the performance of the SMI did not change significantly over the range of tested surface tension (75 – 28mN/m), viscosity (1 – 2.4mPas) and osmolality, thus showing the robust aerosol delivery of the SMI over a relevant range of rheological properties used in inhalation solutions.
Solving AeroChamber alert whistle inconsistencies
Mark Sanders1, Aurelien Martin1 & Darragh Murnane1
1University of Hertfordshire, College Lane, Hatfield, AL10 9AB, UK
Summary: The incidence of inhaler errors, some of which are critical, is high. Spacers are used as a means of reducing throat deposition and mitigating inhaler error. Some spacers such as Aerochamber Plus include an alert whistle to indicate a subject inhaling too fast. In the current study the flow rate at which the Aerochamber Plus whistle was activated was found to vary for different pMDIs, creating the potential to confuse. The range of whistle response was from 30 LMin−1 (Atrovent) to 140 LMin−1 (Flutiform), and is related to device resistance, higher resistance pMDIs sounded at a lower flow rate than low resistance pMDIs. This problem was attributed to the location of the whistle in the chamber where air enters through the pMDI and the whistle, the balance of which is determined by device resistance. The problem can be solved by moving the whistle to the mouthpiece area which then produces consistent whistle activation across all pMDIs.
Evaluating the Impact of a Volume and Flow Resistance Compensator on Flow Rate Rise‐Time Profiles in Dose Uniformity Testing and Aerodynamic Particle Size Distribution Measurement Apparatus for a Dry Powder Inhaler
Ben Bradley1, Clair Brooks1 & Mark Copley1
1Copley Scientific Ltd, Colwick Quays Business Park, Road No 2, Nottingham, NG4 2JY, UK
Summary: Differences in the internal volume of dose uniformity sampling apparatus (DUSA) relative to the cascade impactor set‐ups used for aerodynamic particle size distribution (APSD) measurement are potentially problematic in dry powder inhaler (DPI) testing due to the influence of inspiratory flow rate rise‐time on measured characteristics. A variable volume and flow resistance compensator (VRC) enables the matching of flow rate rise‐time profiles to address this issue, thereby helping to ensure mass balance in compendial APSD measurement methods. Flow rate rise‐time with a DPI DUSA was successfully matched to both the Andersen Cascade Impactor (ACI) and Next Generation Impactor (NGI) to illustrate this approach. The results demonstrate the utility of a VRC for laboratories seeking to minimize sources of variability in DPI testing.
Experimental Evaluations of Internal Losses in ‘Miller’ Mixing Inlet (MI): Part 2 – Dry Powder Inhaler (DPI) Testing for Aerodynamic Particle Size Distribution (APSD)
4Intertek Melbourn, Melbourn, Cambridge, SG8 6DN, UK
Summary: The ‘Miller’ design of mixing inlet (MI) enables a cascade impactor to operate at constant flow rate whilst the inhaler‐on‐test can be evaluated at varying flow rates. We report an evaluation of the losses within this inlet with three separate evaluations; each evaluation conducted with one or more passive DPIs. A similar evaluation with a pressurised metered dose inhaler and nebulizer was presented at DDL‐2022 as Part 1 (Cooper et al.). All evaluations were undertaken with fixed flow rates of compressed air added to the MI to enable a Next Generation Impactor (NGI) to operate at different fixed flow rates without affecting the flow rate from the DPI‐on‐test. The detailed configurations and test conditions are described in the abstract. Internal MI losses were DPI‐dependent, in the worst case, being close to 5%, but more commonly values were in the range from 1 to 4% of the API mass recovered per actuation.
Pulmonary delivery of lysostaphin with a vibrating mesh nebulizer and soft mist inhaler
Ping Zeng1, Pengfei Zhang1, Ho Wan Chan2, Shing Fung Chow2, Jenny Ka Wing Lam2,3, Margaret Ip4 & Sharon Shui Yee Leung1
1School of Pharmacy, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, 999077, China
2Department of Pharmacology and Pharmacy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, 999077, China
3Department of Pharmaceutics, UCL School of Pharmacy, University College London, 29‐39 Brunswick Square, WC1N 1AX, UK
4Department of Microbiology, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, 999077, China
Summary: Methicillin‐resistant Staphylococcus aureus (MRSA) has become a leading causative pathogen of nosocomial pneumonia with an alarming in‐hospital mortality rate of 30%. Last resort antibiotic, vancomycin, has been increasingly used to treat MRSA infections, but the rapid emergence of vancomycin‐resistant strains urges the development of alternative treatment strategies against MRSA‐associated pneumonia. The bacteriolytic enzyme, lysostaphin, targeting the cell wall peptidoglycan of S. aureus has been considered as a promising alternative for MRSA infections. Its proteinaceous nature is likely benefit from direct delivery to the lungs, but the challenges for successful pulmonary delivery of lysostaphin depend on a suitable inhalation device and a formulation with sufficient storage stability. In this study, the applicability of a vibrating‐mesh nebulizer (Aerogen® Solo) and a soft mist inhaler (SMI, Respimat®) was investigated. Both devices were capable of aerosolizing lysostaphin solution into inhalable droplets and caused minimum antibacterial activity loss. In addition, lysostaphin stabilised with phosphate‐buffered saline and 0.1% Tween 80 was proved to have acceptable stability for at least 12 months when stored at 4°C. These promising data encourage further clinical development of lysostaphin for management of MRSA‐associated lung infections.
Assessment of pulmonary delivery of Plasminogen for ARDS local treatment
L. Vizzoni1,2, C. Migone1, B. Grassiri1, F. Mori3, R. Crea3, S. Esin4, G. Batoni4, B. Ferro5, A.M. Piras1
1University of Pisa, Dept. of Pharmacy, Pisa, Italy; 2University of Siena, Dept. of Life Sciences, Siena, Italy
3Kedrion S.p.A., Loc. Bolognana, Gallicano, Lucca, Italy; 4University of Pisa, Dept. of Translational Research and New Technologies in Medicine and Surgery, Pisa, Italy; 5Spedali Riuniti Livorno, Azienda USL Toscana Nord Ovest, Italy
Summary: Acute respiratory distress syndrome (ARDS) is a severe complication of lung injuries, related to bacterial, fungal and viral infections. ARDS is dramatically correlated to patient mortality and no effective treatment is presently available. ARDS involves hypercoagulation, fibrin deposition in the development of an obstructing hyaline membrane, and deep lung inflammation. Plasminogen (PLG) is component of the fibrinolytic system and contributes to inflammation regulatory processes. Pulmonary delivery of PLG, as an off‐label administration of PLG‐OMP eye drops (plasminogen‐based orphan medicinal product) is here proposed as a readily available opportunity for ARDS treatment. The nebulisation was performed using either jet, ultrasonic or mesh nebulisers. Several investigations were performed on the nebulised samples, such as protein aggregation/fragmentation, PLG glycoform distribution, droplets size and aerodynamic distribution as well as enzymatic activity maintenance. Thanks to the excellent efficacy of the mesh aerosolised samples, additional in vitro studies were performed. Mesh nebulised PLG demonstrated good penetration through artificial airway mucus, low cytotoxicity and poor permeation across an Air‐Liquid Interface cellular model of pulmonary epithelium. The results suggest for a good safety profile, excluding high systemic absorption but good mucus diffusion. Most importantly, the aerosolised PLG reversed the effects of LPS‐activated macrophage cells, demonstrating an immunomodulating activity of PLG treatment in an already induced inflammatory state. Repeated‐dose toxicity studies in mice confirmed the tolerability of PLG‐OMP through intra‐tracheal administration. In vivo studies on LPS induced ARDS murine model indicate efficacy in avoiding the most important pathological signs of acute ARDS.
Inhaled biologics: from nebules to SMIs – Dornase alfa in MRX004
Dylan Antoniak1, Philippe Rogeuda1, Adam Stuart1, Graham Purkins1, Chris Vernall2, Daniel Hallisey2, Samuel O'Connell2 & Michelle Lambert2
2Intertek, Saxon Way, Melbourn, SG8 6DN, United Kingdom
Summary: This paper aims to demonstrate that soft mist inhalers (SMIs) are suitable for the delivery of biologics, using dornase alfa nebules (as available in Pulmozyme®) as an example via MRX004 (Merxin Ltd). This testing was performed in collaboration with Intertek and has demonstrated the ability to deliver dornase alfa through a combination of droplet size distribution (DSD) testing via laser diffraction, aerodynamic particle size distribution (APSD) via next generation impactor (NGI), and enzymatic activity via fluorescence tracking. The results show that the MRX004‐produced aerosol has a fine particle fraction (FPF) of 59.4%, with a fine particle dose (FPD) of 125.6 μg. There was only a minimal decrease in enzymatic activity from the native nebule solution, with 90% activity retained, demonstrating the ability of MRX004 to successfully deliver dornase alfa, a biologic. MRX004 is a portable, handheld inhaler that may offer a convenient and effective delivery platform for biologics.
Transitioning pMDIs to SMIs: salbutamol as a case study
Dylan Antoniak1, Philippe Rogueda1, Adam Stuart1 & Graham Purkins1
Summary: There is significant potential benefit in moving from pressurised metered dose inhalers (pMDIs) to soft mist inhalers (SMIs); SMIs are easy to use, may deliver a higher dose to the deep lung and do not require propellant. This paper examines the delivery of salbutamol as a case study to demonstrate how a pMDI can be transitioned to an SMI. It compares the pMDIs Ventolin® (GSK) and Salamol® (Teva) with an SMI – MRX004 (Merxin Ltd) loaded with a commercial 5 mg/mL aqueous salbutamol nebule solution (Aliud). Aerodynamic particle size distribution (APSD) by next generation impactor (NGI), delivered dose (DD), and droplet size distribution (DSD) by laser diffraction were analysed and compared. The results show that the slower plume velocity of MRX004 compensates for its higher median mass aerodynamic diameter (MMAD) – 3.2 μm compared with 2.4 μm for Ventolin® and 2.1 μm for Salamol® – leading to MRX004 to have lower throat deposition, higher fine particle fraction (FPF) and a fine particle dose (FPD) close to that of the pMDIs, despite a much lower total DD than the pMDIs. The FPF was 63.2% for MRX004, 33.6% for Ventolin® and 40.7% for Salamol®; the DD was 56 μg for MRX004, 128 μg for Ventolin® and 108 μg for Salamol®; and the FPD was 35.1 μg for MRX004, 41.4 μg for Ventolin® and 44 μg for Salamol®. MRX004 has a much smaller metering chamber than the pMDIs. MRX004 has a significantly lower deposition in the throat than the pMDIs. When compared to the DSD results, the Dv(50) measurements were 4.7 μm, 2.1 μm and 4.1 μm for MRX004, Ventolin® and Salamol® respectively. This shows that the high‐velocity spray of a pMDI in comparison to the slow mist produced by MRX004 results in impaction that reduces the efficiency of the delivery to the deeper parts of the lungs. These results suggest that MRX004, an SMI, can potentially match or better the performance of pMDIs with additional tuning of the device and formulation. This performance match linked with the lack of propellant makes an SMI option preferable for a low carbon footprint future.
Pathway of Construction and Characterization of a Salbutamol Sulphate HFA152a pMDI Suspension
Giulia Carretta1, Caterina Antiga1, Alberto Colombo2, Giovanni Motta2, Massimo Carrara2, Francesca Buttini1
1Food and Drug Department, University of Parma, Parco Area delle Scienze 27/A, 43124 Parma, Italy.
2RxPack s.r.l, Via Mognago 47, 23848, Oggiono (LC), Italy.
Summary: The work illustrates the development of a suspension of Salbutamol Sulphate (SS) in HFA152a using an open chamber design valve manufactured by RxPack, commercially known as KHFA. Ethanol was used as a co‐solvent and the product target dose was 122 μg of SS (corresponding to 100 μg/shot of salbutamol base). The object of this work was not to make a product copy of the Ventolin® but to develop a good performing product and to be aware of how each factor investigated affects the performance of the system.
Preliminary studies were conducted with a full factorial DoE evaluating the impact of Critical Material Attributes (CMAs), such as ethanol content, valve metering volume, and actuator orifice diameter (AOD), on the critical quality attributes of the products in terms of Emitted Dose (ED) and Fine Particle Fraction (FPF). These initial studies led to identify the lead product consisting of: 50 μL valve, 0.3 mm AOD and 6% ethanol content. The lead product proves to have good respirability thus obtaining FPF values of 45.5% and MMAD of 2.55 μm. So, in conclusion, always paying attention to respecting both the environment (e.g. by making more sustainable choices that lead to less use of propellant and ethanol), and the characteristics that make an inhalation product effective, it is possible to obtain a pMDI with good inhalation characteristics using this new eco‐friendly propellant HFA152a.
Therapeutic effect of an inhaled levodopa dry powder formulation on off episodes in patients with Parkinson's disease
Lara de Jong1,2, Floris Grasmeijer2,3, Marianne Luinstra1, A.F. Aalbers1, L. Wijma‐Vos1, E. D'Angremont4, A.A.E. van der Meulen5, A.W.F. Rutgers5, L. Steenhuis6, P. Hagendoorn2, T. van Laar7 and Erik Frijlink2
1Department of Clinical Pharmacy, Martini Hospital Groningen, Van Swietenplein 1, Groningen, 9700 RM, the Netherlands. 2Department of Pharmaceutical Technology and Biopharmacy, University of Groningen, Antonius Deusinglaan 1, Groningen, 9713 AV, the Netherlands. 3PureIMS B.V., Ceintuurbaan Noord 152, Roden, 9301 NZ, the Netherlands 4Department of Psychiatry, University Medical Centre Groningen, Hanzeplein 1, Groningen, 9713 GZ, The Netherlands. 5Department of Neurology and Clinical Neurophysiology, Martini Hospital Groningen, Van Swietenplein 1, Groningen, 9700 RM, the Netherlands. 6Department of Pulmonary Diseases, Martini Hospital Groningen, Van Swietenplein 1, Groningen, 9700 RM, the Netherlands. 7Department of Neurology, University Medical Centre Groningen, Hanzeplein 1, Groningen, 9713 GZ, the Netherlands.
Summary: Limited treatment options are available with a rapid onset of action to counter off episodes in Parkinson's disease (PD) patients. Therefore, an inhalable formulation of levodopa administered with the single‐use inhaler Cyclops™ is investigated. The pharmacodynamics (efficacy), pharmacokinetics and safety of the levodopa dry powder inhalation formulation was determined in an open‐label crossover trial. Nine participants in an off episode received one dose of inhaled levodopa (90 mg) or one dose (100/25 mg) of levodopa/benserazide orodispersible tablet on two consecutive days in randomised order. A timed finger‐tapping test, Timed Up & Go test and Movement Disorders Society Unified Parkinson's Disease Rating Scale III score were performed predose and on set time points up to 90 minutes postdose as a measure for motor function. In addition, blood samples were taken for a pharmacokinetic evaluation. Lung function was measured as a safety check upon admission and again upon discharge from the clinic. Inhaled levodopa from Cyclops™ had a clinically meaningful effect on improvement of off episodes in PD patients within 20 minutes. The motor function examinations showed a similar clinical improvement after pulmonary and oral administration despite the lower levodopa plasma concentrations after pulmonary administration. Inhaled levodopa was absorbed faster, reaching its peak plasma‐concentration 6 minutes after dosing, as opposed to 28 minutes for oral doses. The levodopa inhalation powder was well tolerated. The overall results of this study suggest that the tested levodopa inhalation powder may be particularly beneficial as an alternative rescue therapy for PD patients with motor fluctuations.
Aerosol Dynamics of Sodium Chloride at High Relative Humidity
Yiliang Lance Jiang1, Gareth Hardwell2, Richard Friend2 & Jonathan P. Reid1
1University of Bristol, School of Chemistry, Cantock's Cl, Bristol, BS8 1TS, United Kingdom
Summary: Inhalers are often prescribed to patients suffering lung or systemic diseases. The size of the particle actuated from the inhalers determines the site of deposition of the therapeutic ingredients. Therefore, an accurate prediction of the drug deposition profile is crucial for maximizing the efficiency of a formulation. Relative humidity (RH) can have a considerable effect on the size distribution of an inhaled medication upon entering the respiratory system. A Tandem Aerodynamic Particle Sizers (TAPS) approach has been developed to measure concurrently the size distribution of the same plume of aerosol at two varied RHs. A Python script was written to fit an optimal size distribution to the experimental data. In this study, sodium chloride have been tested. Comparing the single particle properties, obtained using a comparative kinetics electrodynamic balance (CK‐EDB) with plume dynamics show that they are consistent with predictions from a thermodynamic framework. Further study is required to measure particles with different hygroscopicity and mixtures of particles. Factors such as water content and temperature could also be explored either experimentally or using computational models to further optimize the drug deposition profile.
Development of a Prototype Patient Respiratory Management Mobile Health Application Using Validated Tools to Assess User Experience
Summary: We describe the development of a mobile health (mhealth) application specifically for respiratory disease that would offer a unique opportunity to support individuals with asthma/COPD by providing real‐time monitoring, education, and personalized interventions. The use of the Node.Health and MAUQ validated questionnaires around the areas of usability and ease‐of‐use provides a systematic approach to the iterative development for future versions of the application which will be ultimately become a foundation to which therapeutic devices can be linked to.
Characterization of PARI's new VORTEX® facemasks for babies and children
Andrea Winzen1, Luisa Roßmann1, Yvonne Burmeister2, Paula Riedel2 & Karin Steinführer3
Summary: Two new facemasks were developed for PARI's valved holding chamber VORTEX® to effectively improve aerosol therapy for babies and young children. The mask design was selected to satisfy established requirements for successful inhalation treatment. A detailed analysis of the target group facial anatomy was performed via 3D face scans as basis for geometric and material design choices for the two masks. From the face geometries, basic features were derived and transferred into the contour of the mask. A soft and flexible sealing lip was implemented to allow a tight seal of the mask on children's faces and simultaneously increase comfort during therapy. Dead space volume, transparency, and weight were additionally considered. To evaluate the compliance of the new VORTEX® masks to the defined requirements, measurement of dead space volume and sealing were performed. It showed that the dead space volume was significantly reduced under clinically relevant application forces. Additionally, the leakage rate of the two new masks proved to be very low, (i.e., approximately 2.5% in the entire range of application forces (4 N to 16 N)) indicating a potentially robust drug delivery, even at varying pressures induced by parents on children's faces. This might be beneficial in terms of adherence. For similar reasons, transparency was implemented into the design to allow parents optical access to the children's mouth and nose. The novel VORTEX® masks presented in this study were shown to fulfil major requirements for the effective treatment of babies and children.
On the quest to develop clinically meaningful assessment of the aerosolization performance and respirable dose of nebulization devices
Lois Slator, Jesus Calvo‐Castro & Darragh Murnane
School of Life and Medical Sciences, University of Hertfordshire, College Lane, Hatfield, AL10 9AB, UK
Summary By establishing a test methodology that more accurately emulates the in vivo conditions of aerosol delivery during tidal breathing, a clinically meaningful assessment of the respirable dose may be achieved. This aims to improve the correlation between in vitro and in vivo data, ultimately enhancing the relevance of in vitro testing. The use of the Nephele mixing inlet is proposed as a method to introduce a simulated breathing pattern into test systems for aerosol particle size diameter, currently only measured under continuous extraction. Previously published testingi indicated an observed decrease in the fine particle dose from a continuous mesh nebuliser was due to a decrease in the amount of drug delivered rather than a change in particle size. The same test setup has been used to evaluate the fine particle fraction and ex‐throat drug dose collected and calculate the fine particle dose for three more types of nebuliser; a continuous jet, a breath‐enhanced jet and a breath‐activated mesh nebuliser. The fine particle dose of the breath‐enhanced jet nebuliser was most affected by the simulated breathing test set‐up, with the ex‐throat collected dose reduced from 377 to 239 μg, although the fine particle fractions were similar (60.4% and 57.1%). This resulted in a decrease in fine particle dose from 227 μg under continuous conditions to 136 μg under simulated breathing conditions.
Feasibility Development of Salbutamol Sulphate Pressurized Metered Dose Inhalers using a Low Global Warming Potential Propellant 1,3,3,3‐Tetrafluoropropene (HFO1234ze)
Lei Mao, Nischal Pant, Danielle Sousa, Emily Humphreys, Danny Anderson, Ben Rinne
Recipharm Laboratories, Morrisville, North Carolina, 27560, USA
Summary: A feasibility study was conducted to develop salbutamol sulphate (SS) pressurized metered dose inhalers (pMDIs) using a low global warming potential (GWP) propellant 1,3,3,3‐Tetrafluoropropene (HFO1234ze) and novel Bespak valves developed by Recipharm to match Ventolin® performance. The formulations containing SS and HFO1234ze with and without ethanol (2%) were prepared manually using coated aluminium cans. The pMDIs were tested along with Ventolin® for through life delivered doses (DD), aerodynamic particle size distribution (APSD), spray pattern (SP), plume geometry (PG), assay, impurities, moisture and leak rate. The formulation containing 2% ethanol gave a consistent DD through inhaler life but a high induction port and low stage deposition when tested using the Next Generation Pharmaceutical Impactor (NGI). Formulations without ethanol showed variable DD through inhaler life. Investigation is ongoing to understand whether this was attributed to the formulations or inhaler handling during testing. NGI results showed that the formulations without ethanol gave the same percentage deposition on the induction port but slightly higher stage 3 and 4, and lower stage 5,6,7 deposition which can be corrected by adjusting the particle size of the micronized SS. The PG matched well to that of Ventolin® but the SP, i.e. Damax, Dmin and area were slightly smaller which can be rectified by modifying the actuator. Two novel valves showed a lower leak rate compared to the generic one used in the current propellent. The formulation containing 2% ethanol had a higher moisture content. No impurities were detected and SS assay values were close to the input. The formulations are being evaluated for stability.
Overview of the Nonclinical Safety Program for 1,1‐Difluoroethane (HFA‐152a) as a Low Global Warming Potential Metered Dose Inhaler Propellant
1Gradient, 600 Stewart Street, Suite 1900, Seattle, WA, USA
2Koura, Thornton Science Park, Pool Lane, Ince, Chester, CH2 4NU, United Kingdom
3Independent Consultant, Tijeras, NM, USA
Summary: Pressurized metered dose inhalers (pMDIs) deliver life‐saving medicines with more than 800 million pMDIs produced annually. 1,1‐Difluoroethane (HFA‐152a) is being developed as a pMDI propellant and as a more environmentally sustainable alternative to current hydrofluoroalkane (HFA) propellants, with an approximate 90% reduction in global warming potential. HFA‐152a is a gas at ambient temperature and pressure, has no intended or expected pharmacologic activity, and has been shown to be well tolerated and without adverse effects in a Phase 1 clinical study of HFA‐152a administered via a pMDI. In this paper, we summarize the results of a comprehensive nonclinical safety program for HFA‐152a to support clinical use in pMDIs. The nonclinical safety program showed that medical‐grade HFA‐152a caused no toxicity in mice, rats, or dogs, and no carcinogenicity in rats at delivered doses a minimum of 1800 times the anticipated clinical maximum daily exposure (MDE) for adults. HFA‐152a caused no developmental or reproductive toxicity in rats or toxicity in juvenile rats at delivered doses greater than 2400 times the MDE. Furthermore, the results of specialized safety pharmacology studies showed that clinical use of HFA‐152a would not pose a meaningful risk of respiratory sensitization or cardiac sensitization. Lastly, the results of bacterial mutagenicity, in vitro micronucleus, and in vivo micronucleus assays did not show HFA‐152a to be mutagenic or genotoxic. Overall, the current nonclinical safety program for medical‐grade HFA‐152a supports the safety of HFA‐152a as a pMDI propellant for inhaled drugs intended for all patient populations and chronic daily use.
Spray Pattern and Plume Geometry Measurements of Current and Low GWP Propellants HFA152a and HFO1234ze(E) pMDIs With and Without Ethanol
Holly Dowdle1, Benjamin Myatt1
1Kindeva Drug Delivery, Charnwood Campus, Loughborough, UK
Summary: As the pressurized metered dose inhaler industry is making the switch to lower global warming potential propellants, more understanding is needed on how the new propellants perform in pMDI products using existing characterisation techniques. This study characterises the spray pattern and plume geometry of propellant and propellant with 12% w/w ethanol formulations of the current and new propellants. Standard spray pattern tests at 30 mm, 60 mm and plume geometry in line with industry guidance were performed using an Oxford Lasers EnVision Pharma R&D system. These tests revealed that both HFA134a and HFA152a had the largest spray pattern areas at 30 mm and 60 mm, with HFA227ea having the smallest area, due to the differences in propellant saturated vapour pressure. The spray pattern areas were proportional to vapour pressure at 30 mm distance. However, for HFA152a the plume expanded at a greater rate than that of HFA134a, which could be due to the lower density of HFA152a. When adding ethanol to the propellants, the spray pattern area reduced for most of the propellants and the shot‐to‐shot variability reduced; this would indicate that adding ethanol stabilises formulations. These results present the need for formulators to understand more about the physical properties of the propellants, so that similar product performance can be achieved when it comes to reformulating with the new propellants.
Importance of the Tongue as an Impactor for Orally Inhaled Aerosols from a Pressurised Metered Dose inhaler (pMDI) With and Without a Valved Holding Chamber (VHC)
Mark W Nagel1, Cathy Doyle1, Jason A Suggett1 & Jolyon P Mitchell2
2Jolyon Mitchell Inhaler Consulting Services Inc., 1154 St. Anthony Rd., London, Ontario, N6H 2R1, Canada
Summary: We undertook a study comparing fine particle fraction (FPF<4.7μm) and fine particle mass <4.7 μm aerodynamic diameter (FPM<4.7μm) from a pMDI delivering salbutamol (100 μg/actuation) with and without a VHC. A series of four model adult oropharyngeal cavities (OCs) were used, in which the tongue size was progressively reduced in volume (100%, 60%, 30% and 0%). FPF<4.7μm for the pMDI alone increased significantly (1‐way ANOVA, p < 0.0001) with reduced tongue volume, from 16.7 ± 3.6% with the full tongue volume present, to 38.5 ± 6.5% when the tongue was completely removed. FPM<4.7μm increased from 12.9 ± 2.1 μg/actuation to 26.5 ± 3.8 μg/actuation (p < 0.0001). When the VHC was present, with a 2 s delay between actuation and inhalation the FPF<4.7μm remained relatively consistent across the different tongue volume conditions, and the FPM<4.7μm changed much less than with the MDI alone. We confirmed the importance of the tongue in controlling fine particle delivery beyond the oropharynx. When the VHC was absent, the data confirmed clinical guidance to insert the mouthpiece above the tongue before actuating and simultaneously inhaling the medication. If a VHC is present, we demonstrated consistent fine particle delivery irrespective of tongue volume, without the need to coordinate actuation with inhalation. Such changes in tongue position may better reflect the variability in human anatomy in inhaler use.
SoundCatcher: Prediction of Inhalation Profiles and MDI Actuation Synchronization using Acoustic Emission with Machine Learning
Ellinor Nilsson1, Mats Josefson2, Lisa Holmstén1, Patrik U. Andersson1, Lubomir Gradinarsky3, Johanna Fredriksson3 and Lars Karlsson1
3Innovation Strategies & External Liaison, Pharmaceutical Technology and Development, AstraZeneca, Gothenburg, Sweden
Summary: The combination of acoustic emission (AE) and machine learning (ML) has been shown to be a powerful tool in quality control related analysis as well as in patient adherence monitoring. In this study it has been used for monitoring of inhalation profile and of synchronization between actuation and inhalation of MDI devices. In this set‐up, pMDIs were used as model devices. For the inhalation profile experiments a lung simulator was used for simulation of adults' weak, medium and strong inhalation profiles. For the synchronization experiments a constant flow was used and device actuation was performed at different time points in relation to the flow onset. To challenge the ML algorithm, the experiments were also performed in a noisy environment to mimic a domestic environment. An image‐based ML model with Convolutional Neural Networks (CNN) was used to predict the inhalation profile and the synchronization between actuation of device and inhalation. By this approach it was possible to obtain accurate predictions
Optimising Actuator Geometries of Low GWP Propellant HFA‐152a Solution Formulations
Ameet Sule1, Sunita Sule1, John Howard1, Nisar Ahmed1, Robert Bootle1
1H&T Presspart Inhalation Product Technology Centre, Whitebirk Industrial Estate, Blackburn BB1 5RF, UK
Summary: Low Global Warming Potential (GWP) propellants are being explored for pressurised metered dose inhalers (pMDIs) to reduce the carbon footprint. The selection of propellant in a metered dose inhaler is a substantial change to the product that will require not only a good deal of consideration with regards to the formulation, but also the device componentry for example actuators.
Actuator geometries are critical parameters which influence the shape and size of the aerosol plume and can significantly affect the aerodynamic drug distribution, thereby affecting the overall efficacy of the product for the patient.
Through spray pattern testing, the plume characteristics are used as a comparative tool against other actuator configurations, including a reference device (RLD) and can help predict in‐vitro device performance.
The performance of three different solution formulations, with HFA‐152a as the propellant and ethanol as a co‐solvent, with different actuator geometries were evaluated by assessing the spray behaviour at two distances1. The results were compared against commercially available products with HFA‐134a formulations. Several actuator geometry configurations showed comparable performance to the RLD at one distance (p ≥ 0.05, n = 15); however, no configuration tested, showed p ≥ 0.05 at both distances.
Understanding fundamental formulation properties to demonstrate the feasibility of reformulating a salbutamol pMDI containing the low GWP propellant HFO1234ze(E)
Alex Slowey, Phil Cocks and Ben Myatt
Kindeva Drug Delivery, Charnwood Campus, Loughborough, UK
Summary: The most widely used salbutamol sulphate rescue medications are pMDIs containing propellant HFA134a, which has a global warming potential (GWP) of >1400. It is therefore economically and environmentally desirable to develop new pMDI products, including rescue medications, using low GWP propellant alternatives.
Kindeva Drug Delivery (Kindeva) has demonstrated that a low GWP salbutamol pMDI product could be developed that achieves comparable in‐vitro pharmaceutical performance to Ventolin™ Evohaler™. The propellant chosen for reformulation was HFO1234ze(E) primarily due to its low GWP of <1.
Initial investigations demonstrated that it was not possible to just change the propellant from HFA134a to HFO1234ze(E) due to high levels of drug deposition on the internal surfaces of the container closure system (CCS), which resulted in a low and variable delivered dose relative to Ventolin™ Evohaler™.
After the inclusion of small quantities of ethanol in the alternative product formulation, delivered dose was significantly improved and comparable DCU performance to Ventolin was achieved. However, it was recognised that differences in the properties of the two propellants and the inclusion of ethanol, would significantly affect atomisation behaviour and plume evolution of the reformulated test product.
Based on a fundamental understanding of the thermophysical properties of both propellants, in particular, HFO1234ze(E)/ethanol binary mixtures, key actuator dimensions (exit orifice diameter and jet length) were selected to enable the APSD performance of the reformulated test product to match that of Ventolin. The test product was confirmed to be physically and chemically stable following 13 weeks storage at accelerated conditions (40°C/75% relative humidity).
In conclusion, this study successfully demonstrated that a reformulated salbutamol sulphate pMDI product with low GWP propellant HFO1234ze(E), containing 5% w/w ethanol, resulted in comparable in‐vitro pharmaceutical performance to Ventolin Evohaler.
Device and formulation parameters influencing pressurised Metered Dose Inhaler (pMDI) aerosols using HFO1234ze propellant
Thorne, B. J. A.1, Vaccaro, G.2, Cotton, D. P. J.1, Kirton, S. B.2, Knowles, M.2, Lee, K. C.3, Murnane, D.2, Sapsford, A. I.1
1Recipharm, Bergen Way, King's Lynn, PE30 2JJ, U. K., 2University of Hertfordshire, College Lane, Hatfield, Hertfordshire, AL10 9AB, 3University of East London, Docklands Campus, University Way, London, E16 2RD
Summary: Next generation impactor (NGI) stage‐by‐stage performance was measured for two pressurised Metered Dose Inhaler (pMDI) device configurations for an ethanolic fluticasone propionate (FP) formulation using the low global warming potential (GWP) propellant HFO1234ze. This was compared with ex‐actuator aerosol droplet size distribution measurements captured using a Malvern Spraytec laser diffraction system for an FP solution formulation. The droplet size distributions immediately ex‐actuator were also predicted using our 1‐dimensional thermofluid mechanic model for both formulations. The simulation predicted only a minimal influence of ethanol on the initial droplet sizes, whereas actuator spray orifice diameter was a much more significant variable. The NGI data showed significant throat deposition for the solution formulation used in conjunction with the larger actuator spray orifice diameter, with a commensurate reduction in subsequent stage deposition. This suggests that the droplets received at the NGI throat had not evaporated fully. The laser diffraction data indicated incomplete evaporation of the solution emitted droplets at a distance of 150 mm from the pMDI actuator. An additional laser diffraction assessment of an FP suspension aerosol indicated complete evaporation of propellant by 150 mm. This highlights the importance of droplet evaporation kinetics on the ultimate destination of the dose in the body.
Physical Stability of Salbutamol Sulfate Suspensions in HFA‐134a and HFO‐1234ze(E)
Erik Boldt1, Gregory Smith1
1Honeywell International, 20 Peabody Street, Buffalo, NY 14210, USA
Summary: With the growing push to convert pressurized metered dose inhalers (pMDIs) to new low global warming potential (GWP) propellants, an understanding of the physical compatibility of active pharmaceutical ingredients (APIs) with the propellants is necessary for reformulation. Suspension formulations encounter several challenges that solution formulations do not, including destabilization through sedimentation, creaming, flocculation, and/or coalescence. Each of these can have negative effects on the formulation, causing performance issues. Therefore, a thorough understanding of the physical suspension is of the utmost importance.
This study examined the physical stability of salbutamol sulfate formulations in both a currently used propellant, hydrofluoroalkane (HFA)‐134a, and an alternative low‐GWP propellant hydrofluoroolefin (HFO)‐1234ze(E) through use of static multiple light scattering (SMLS) methods. Salbutamol sulfate suspensions in HFO‐1234ze(E) were more stable and had a lower sedimentation rate compared to HFA‐134a, though flocculation was more persistent. The addition of ethanol reduced flocculation and sedimentation rate in HFO‐1234ze(E), increasing the total stability over time.
Stability Evaluations of Salbutamol Sulphate Propellant‐Dispersible Tablets (Respitab™) for Formulation with HFA‐152a
Summary: Respitab™ is a novel approach for manufacturing pressurised metered dose inhalers (pMDIs) using propellant‐dispersible tablets. In this production process a tablet is dispensed into the canister, the metering valve crimped and the propellant pressure filled. This innovation circumvents manufacturing challenges associated with flammable gasses e.g. hydrofluoroalkane 152a (HFA‐152a) by reducing the number of essential operator interventions associated with production and cleaning during traditional single‐stage or two‐stage filling. Dispensing propellant‐dispersible tablets into canisters and crimping valves, independently of gassing provides greater flexibility in the manufacturing process. For example, active pharmaceutical ingredient (API) handling can be performed separately from propellant filling which as a consequence may be greatly simplified enabling significant savings in terms of investment in manufacturing equipment and modification of production facilities.
In this study Respitab propellant‐dispersible tablets containing salbutamol sulphate (SS) were dispensed into plain aluminium canisters and metering valves were crimped to seal. Canisters were subjected to accelerated stability conditions (up to 3 months at 40°C/75% RH, no foil overwrap) prior to addition of HFA‐152a. Following gassing, aerosol characteristics i.e. fine particle fraction (FPF) and dose content uniformity (DCU) through canister life were determined. The moisture level within the formulation was also assessed.
FPF and DCU data were consistent over 3 months' storage. DCU showed the mean metered dose was within acceptable limits i.e. all within ±35% of target and 90% of results within ±25% of target. FPF indicated consistent and efficient aerosol generation, with mean FPF values ranging from 62% to 66%.
The Effect of Elastomer Type on Stem‐Diaphragm Leakage from pMDIs filled with Propellant 152a
Howard Bleazard, Lee Hodges
Kindeva Drug Delivery Ltd, Upbrooks, Clitheroe, BB7 1NX
Summary: Leakage from a pressurised metered dose inhaler (pMDI) comes from two sources: the valve‐to‐canister crimp region and the metering valve's stem‐diaphragm interface. Both regions of the pMDI typically use elastomeric components to create an effective seal. Whilst the valve‐to‐can crimp is a static seal the stem‐diaphragm interface is dynamic, in that the stem slides through the diaphragm each time the pMDI is actuated.
Initial evaluation of propellants 152a and 1234ze on pressure fill metering valves designed for use on propellants 134a and 227 has shown a much‐increased leakage rate, especially so with propellant 152a. In the study summarised below, an evaluation of the effect of elastomer material on stem‐diaphragm leakage has been performed using placebo formulations comprising propellant 152a with and without ethanol. Three types of elastomer materials have been evaluated: nitrile, EPDM and chlorobutyl under two temperature regimes. Weight loss was determined by weighing units on an analytical balance initially and then again after increasing time periods (in a temperature‐controlled environment) ranging from 7 days to 6 months. An annual leakage rate was calculated by linear extrapolation.
Results indicate a clear difference in stem‐diaphragm leakage dependent on the type of elastomer used to manufacture the diaphragm component; in the worst‐case scenario of a 40°C storage regime and units filled with propellant 152a and 10% (w/w) ethanol, maximum leakage from a pMDI with a chlorobutyl diaphragm was 56 mg/year, compared to 225 mg/year with an EPDM diaphragm and 1076 mg/year with a nitrile diaphragm.
The delivery of CRISPR‐Cas9 targeting KRAS mutant in lung cancer
1Ludwig‐Maximilians‐University, Department of Pharmacy, Pharmaceutical Technology and Biopharmaceutics, Butenandtstraße 5‐13, Munich, 81377, Germany
Summary: CRISPR‐Cas9 has been proven to be an efficient and customizable genome editing tool that can be potentially used for the treatment of KRAS mutation in lung cancer. Here we report a cationic polymer system, namely lipid modified polyethylenimine (C14‐PEI), that can co‐deliver Cas9 mRNA and sgRNA efficiently to excise the mutated KRAS allele in lung cancer. C14‐PEI is prepared from epoxytetradecane and branched PEI 600 Da in a ring opening reaction. C14‐PEI shows a critical micelle concentration (CMC) at around 21 mg/L and can efficiently encapsulate mRNA as micelleplexes through electrostatic interaction. eGFP mRNA can be encapsulated by C14‐PEI with 99% encapsulation efficiency at the mass ratio of 8 (w/w 8), and it showed 130‐fold increased expression in A549 cells compared with the PEI control treatment group. And no significant toxicity was observed through the test of intracellular reduction of a tetrazolium salt. C14‐PEI showed high efficiency to co‐encapsulate Cas9 mRNA and sgRNA together based on agarose gel electrophoresis analysis. At the sgRNA to Cas9 mRNA molar ratio 10, the micelleplexes successfully mediate KRAS mutant cutting with 63% indel efficacy tested by T7EI
A New Small Molecule Effective Against Common Pathogenic Respiratory Viruses with Characteristics Optimal for Direct Delivery to the Respiratory Epithelium
David P. Fergenson1, Li Du2, David Hanzel3, Homer Boushey3,4, Jim Lin3, Maya Michon3, Anuradha F. Lingappa3, Kumar Paulvannan3, Shao Feng Yu3, John R. Greenland4, Satish Pillai2,4, Vishwanath R. Lingappa3,4 & Igor Gonda3
1Livermore Instruments Inc., 2038 Livingston Street, Suite B, Oakland, CA 94606, USA
2Vitalent Research Institute, 360 Speer Street, San Francisco, CA 94112, USA
3PRAV Biosciences, Inc.,670 5th Street, San Francisco, CA 94107, USA
4The University of California, San Francisco, CA 94143, USA
Summary: We present our in vitro finding that PAV‐104, a novel broad‐spectrum small molecule antiviral, is ideally administered to the apical surface of the lung epithelium as in the case of an inhaled formulation. The PAV‐104 chemotype exhibits nanomolar activity against the six viral families that cause 95% of human respiratory viral disease including SARS‐CoV‐2; shows minimal toxicity in rats at a dose achieving >100x EC50 blood levels for 24h with good lung exposure; and appears not to generate resistance in vitro [2]. In vitro evidence suggests that the broad effectiveness and low toxicity result from PAV‐104 targeting an allosteric site on a subset of the protein 14‐3‐3 that is retasked from an autophagy role in healthy cells to a viral replication role by many viruses; PAV‐104 appears to target only the disease‐associated conformation of the protein. Compounds of the PAV‐104 chemotype have shown efficacy in two animal models [2]. Given the obvious utility of a pan‐respiratory anti‐viral compound without the liability of resistance development, we sought to identify its optimal mode of administration. Here we show that the application of PAV‐104 at 50nM to the apical surface of primary human bronchial epithelial cells cultured at an air‐liquid interface (HBE@ALI) reduced viral load by >3 logs, significantly more than the 1 log drop achieved upon dosing at the same concentration from the basal surface. These findings suggest that PAV‐104 is a prime candidate for inhalation delivery.
Pulmonary Delivery of Hybrid Polymeric Nanoparticles for Enhanced mRNA Expression
Min Jiang1, Felix Sieber‐Schäfer1, Simone P. Carneiro1, Olivia M. Merkel1,2*
1Department of Pharmacy, Ludwig‐Maximilians‐University Munich, Butenandtstrasse 5‐13, Haus B, 81377 Munich, Germany
2Center for NanoScience (CeNS), Ludwig‐Maximilians‐University Munich, 80799 Munich, Germany
Summary: Recent advances in the field of mRNA vaccination have demonstrated significant advantages in activating immune responses and combating infectious diseases. However, many pathogens and viruses, including SARS‐CoV‐2, primarily infect the host through the respiratory tract. Conventional vaccines administered via non‐respiratory routes usually fail to induce protective mucosal immunity, which limits the elimination of pathogens and viruses at the first entry site and still results in the risk of acute infection. Therefore, we aimed to develop a pulmonary mRNA nanovaccine capable of achieving both crucial mucosal immunity and systemic immunity against infectious diseases. To accomplish this, we have innovatively designed a hybrid nanoplatform based on a non‐lipid system, integrating biodegradable poly(lactide‐co‐glycolide) (PLGA) and poly (β‐amino esters) (PBAEs) into nanoscale core‐shell particles for loading and delivering mRNA. The mRNA‐loaded hybrid nanoparticles had an average size of 198.8 ± 4.4 nm and exhibited a slightly positive charge in 10mM pH7.4 HEPES buffer. Through evaluation of the post‐transfection effects of eGFP‐mRNA on both dendritic cells and lung epithelial cells, we have confirmed the superiority of the nanohybrid system in mRNA delivery compared to the PBAE‐based nanopolyplex system used alone. This enhanced delivery efficiency can be attributed to the optimized structure, enhanced hydrophobicity and improved stability of the hybrid nanoparticles, which will be further explored in future research.
Aerosol Characterization, Toxicokinetic and Biodistribution Investigations Supporting Efficient Delivery of KIT2014 Peptide to the Airways in Mice after Nose‐only Inhalation
Valentina Sala1 & Blane Stobbs2
1Kither Biotech Srl, Via Nizza 52, 10126 Torino, Italy
2Charles River Laboratories Edinburgh Ltd, Elphinstone Research Centre, EH33 2NE Tranent, United Kingdom
Summary: Proof‐of‐concept studies in vitro and in vivo have shown the therapeutic potential of KIT2014, a phosphoinositide 3‐kinase gamma mimetic cell‐permeable peptide, in the context of chronic obstructive respiratory diseases. Therefore, in accordance with international guidelines, inhalation studies in relevant species are needed to define the safety profile of KIT2014, to progress towards clinical development. This study enabled set up of the appropriate conditions to generate and characterize for the first time the aerosol features, biodistribution and bioavailability of respirable KIT2014 aerosol, suitable for nonclinical safety studies. The aerosol was generated by vibrating mesh nebulisers and administered to mice by nose‐only flow past exposure system. Study results were pivotal to ensure feasibility of in vivo inhalation studies, and paved the way to follow‐on GLP repeated dose studies.
Spray drying of mRNA lipoplexes to produce an inhalable dry powder formulation for mRNA therapeutics
Jana Schembera1, Ricarda Leister2, Constanze Müller2 and Regina Scherließ1
1Department of Pharmaceutics and Biopharmaceutics, Kiel University, Gutenbergstrasse 76, 24118 Kiel, Germany
Summary: This project aims to develop an inhalable dry powder platform for mRNA therapeutics. To create this dry powder formulation, we utilised lipoplexes containing mRNA and DOTAP:DOPE and conducted spray drying experiments using mannitol as a matrix. Lower outlet temperatures preserved transfection efficiency better than higher outlet temperatures. After redispersion of the spray dried powder, the z‐average and zeta potential indicated stable nanoparticles within the desired size range for all used outlet temperatures. The aerodynamic characterisation of the spray dried powder showed a moderate fine particle fraction (approx. 35%) using the RS01 DPI as device. However, a considerable amount of powder remained in the capsules and the throat. In a first attempt, lactose sweeper crystals were introduced as a potential solution to enhance particle detachment. The sweeper crystals did not affect the fine particle fraction in this study, while they tend to increase the delivered dose. Further research is required to improve the composition of the sweeper crystal blends and explore alternative strategies to tune aerodynamic performance.
Inhaled ceftazidime dry powder developed using spray freeze drying technique
Rishi M. Shah1,2, Anlin Lyu1, Greg F. Walker1, Rajesh Katare2,3 & Shyamal C. Das1
1School of Pharmacy, University of Otago, Dunedin 9054, New Zealand.
2Department of Physiology, University of Otago, Dunedin 9016, New Zealand.
3Department of Physiology, HeartOtago, School of Biomedical Sciences, University of Otago, Dunedin 9016, New Zealand.
Summary: Pulmonary delivery is an attractive approach to the management and treatment of lung infections. The delivery of antibacterial agents directly to the lungs provides a high drug concentration to the site of action and reduces systemic toxicity. However, the number of approved inhalation antibiotics is minimal, necessitating the urgent need for new inhaled antibacterial compounds. The reformulation of existing drugs is a promising way to develop new inhaled therapeutics. Ceftazidime, a potent broad‐spectrum antibiotic used to treat bacterial infections, was used as the model drug. This molecule exhibits poor aerosolisation properties and degradation. Among the several methods implemented for producing inhaled formulations, spray‐freeze drying has recently gained significant attention as a promising formulation approach which can enhance the aerosolisation of ceftazidime formulation. Spray‐freeze drying is a hybrid technique that produces highly porous and dispersible dry powders with relatively low aerodynamic diameters that are suitable for pulmonary administration. L‐leucine was employed as an aerosolisation enhancer. The formulations were characterized using scanning electron microscopy, infrared spectroscopy, water content, drug content and in vitro aerosolisation performance. SEM analysis suggests that the use of L‐leucine resulted in highly porous spherical particles. The powder had ⁓9% water content and a drug content of 94%. It was found that adding L‐leucine to the ceftazidime formulation enhanced its fine particle fraction by 1.2‐fold. This study demonstrates the advantage of L‐leucine in improving the aerosolisation performance of spray‐freeze‐dried particles.
Anti‐biofilm activity of aerosolized phage vs. ciprofloxacin using an in vitro lung deposition platform
1McMaster University, Hamilton, ON, L8S 4L8, Canada
2Department of Chemical Engineering
3Faculty of Health Sciences, Department of Medicine
4St. Joseph's Healthcare, Firestone Research Aerosol Laboratory, Hamilton, ON, L8N 4A6, Canada
5School of Biomedical Engineering
6Michael DeGroote Institute for Infectious Disease Research
7Farncombe Family Digestive Health Research Institute
Summary:Pseudomonas aeruginosa is a bacterial pathogen frequently implicated in recurring, antibiotic‐resistant airway infections. Aerosolized treatments for lower respiratory tract infections should include fine droplets <5 μm in aerodynamic size that can effectively penetrate and deposit at distal sites within the lungs. An in vitro platform composed of a six‐stage viable cascade impactor fitted with transwell inserts containing P. aeruginosa biofilms was used to model different sites of infection within the airways. Biofilms were subsequently exposed to size‐fractioned droplets of nebulized bacteriophage (phage) or the fluroquinolone antibiotic ciprofloxacin. Exposure to phage, regardless of the dose delivered by size‐fractioned droplets, resulted in significant reduction of free‐floating and adherent bacteria as compared to negative control samples. Conversely, antibacterial efficacy of ciprofloxacin exhibited a strong dose dependence. Only biofilms that received the highest dose of ciprofloxacin of 2.5 μg, delivered by ≥7 μm sized droplets, exhibited significant reduction in viability as compared to negative control samples. The superior performance of phage may be attributed to its self‐replicating nature, allowing for on‐site amplification from repeated cycles of infection, propagation, host cell lysis, and progeny virion release. Lacking this characteristic, intervention with nebulized ciprofloxacin proved to be dose restrictive. This study demonstrates the promise of inhaled delivery of lytic phages for tackling persistent deep lung infections as compared to conventional antibiotics.
Molecular Size of Linear and Branched Polyethylemine Influences the Transfection Efficacy of mRNA Polyplexes Aerosolised using a Vibrating Mesh Nebulizer
Abhimata Paramanandana, Hao Ying Li & Ben Forbes
King's College London, Institute of Pharmaceutical Science, London SE1 9NH, UK
Summary: Messenger ribonucleic acid (mRNA) therapeutics have gained considerable attention in recent years due to their wide range of potential medical applications. However, they face challenges in terms of drug delivery; notably their cellular uptake and stability. In this study, the production of a reporter mRNA sequence was optimised and the stability of nanoparticle polyplexes to aerolisation was evaluated by measuring transfection in cells. The goal was to explore the influence of delivery vectors on the stability and the efficiency of respirable mRNA therapeutics. Delivery by inhalation offers advantages for nucleic acid delivery including direct lung targeting and bypassing metabolic and other clearance mechanisms. The study used green fluorescent protein (GFP) mRNA as a model nucleic acid therapeutic and utilized linear and branched polyethyleneimine (PEI) polymers of different molecular weight for nanoparticle formulation. Plasmid DNA linearization was achieved using AfeI and PmeI restriction enzymes and in vitro transcription was effected using a MAXIScript™ Kit. Gel retardation assays confirmed effective mRNA complexation within the PEI polyplexes and evaluated physical polyplex stability after nebulization. Transfection studies on HEK 289T cells showed successful uptake of the polyplexes and expression of GFP. This study demonstrated that it is possible to formulate mRNA polyplexes that are physically stable to aerosolization using a vibrating mesh nebuliser, and used cell transfection to show that their biological effects are retained but diminished. These findings highlight the potential of inhalation‐based delivery as a promising strategy for lung‐targeted mRNA therapeutics.
Increases in Lipid Nanoparticle‐mRNA Particle Size upon Mesh Nebulisation Vary by Aerodynamic Fraction
Hao‐Ying Li1, Bhavini Savarn1, Luke Granger2, Francois Hallac3, Muattaz Hussain4, Abhimata Paramanandana1, Liya Chiu5, Yvonne Perrie4, Robin Shattock2, Charalampos Makatsoris3, Ben Forbes1
1Institute of Pharmaceutical Science, King's College London, London SE1 9NH, United Kingdom
2Department of Infectious Diseases, Section of Immunology of Infection, Imperial College London, London W2 1PG
3Department of Engineering, Faculty of Natural & Mathematical Sciences, King's College London, London WC2R 2LS
4Strathclyde Institute of Pharmacy and Biomedical Sciences, The University of Strathclyde, Glasgow G4 0RE
5Centillion Technology Ltd., 153 Cambridge Science Park, Cambridge CB4 0GN
Summary: In this study, messenger ribonucleic acid (mRNA) was synthesized by in vitro transcription (IVT) and used to prepare lipid nanoparticles (LNP) by combining the charged nucleic acid and lipid vector in a 1:8 ratio (N/P = 8). The LNP‐mRNA non‐viral delivery system was investigated for its suitability for mRNA delivery to the lungs by using a vibrating mesh nebuliser to aerosolize the LNP‐mRNA formulation. It was found that, in comparison with their properties before aerosolisation, the aerosolized LNP‐mRNA demonstrated increased particle size (120.3 nm vs. 211.1 nm), higher surface charge (6.12 mV vs. 13.94 mV) and enhanced encapsulation efficiency (EE, 90.9% vs. 99.9%). Collection and analysis of the aerosol fractions impacted on stages 2‐6 of Next Generation Impactor (NGI) showed the nanoparticles recovered from the later stages had increased particle size, while there was no obvious difference in the zeta potential and EE for LNP‐mRNA on different stages. Overall, these data showed that the aerosolized LNP‐mRNA had an average particle size of ∼200 nm, surface charge of ∼+13 mV, and EE >90% w/w. The vibrating mesh generated respirable aerosol droplets for pulmonary delivery of LNP‐mRNA, with a fine particle (< 5.0 μm) fraction of 49.3 % w/w and mass median aerodynamic diameter of 5.1 μm.
Evaluation of impurities, degradation products and foreign particles arising in an unwashed valved holding chamber (VHC) used with a pressurised metered dose inhaler (MDI)
Justine Bloch1,2 & Yannick Baschung1
1Drug Delivery and Physical Characterization, Solvias AG, Kaiseraugst, Switzerland
2Department of analytical chemistry, Université Claude Bernard Lyon 1, Lyon, France
Summary: We aimed to investigate the potential accumulation of impurities and degradation products that could arise from the misuse of a pressurised MDI (Symbicort® Rapihaler® Budesonide/Formoterol 160 ug/4,5 ug, AstraZeneca) when used in combination with a VHC (AeroChamber Plus®, Sandoz). To simulate real‐life scenarios, we subjected the spacer to six actuations per day over a three‐month period without any cleaning. Following this simulated usage, we distributed two doses in a collection tube through the VHC and extracted all components to identify any impurities. A noticeable increase in budesonide, formoterol and their degradation products were detected both in the VHC and in the collection tube during the extraction process. This finding emphasizes the importance of regular cleaning and maintenance to prevent substantial build‐up over time. The notable increase in impurities and APIs levels within the collection tube raises possible concerns about the potential impact on patient safety when cleaning is neglected for an extended period. These results underscore the necessity for vigilant cleaning practices and highlight the limitations of the VHC in preventing impurity transfer over extended periods of misuse and provide valuable insights into the behavior of MDI spacers under misuse conditions.
From formulation physicochemical properties to digital next generation impactors
Daniel Hardy1, Jianghan Tian1 & Jonathan Reid2
1Microsol, 20 Wenlock Road, London, N1 7UG, UK
2School of Chemistry, University of Bristol, Bristol BS8 1TS, U.K
Summary: Simulations of next generation impactor (NGI) deposition were performed in‐silico, utilising state‐of‐the‐art aerosol kinetics models and single‐particle measurements of hygroscopicity. Particle size evolution was captured for full aerodynamic particle size distributions (APSDs), and coupled with analytical models of filtration systems. This methodology was demonstrated for a soft mist inhaler aerosol with the potential to be applied to any formulation. Literature descriptions of inertial impactors were used to generate digital representations of an NGI. Experimental data of sodium chloride, salbutamol sulphate and tobramycin were used for the simulation of aqueous aerosols. The effect of formulation properties, in particular, hygroscopicity was shown to have an impact upon the size evolution of each aerosol. The effect of the gas phase relative humidity (RH) upon aerosol distributions was shown as was the corresponding effect upon NGI deposition. A larger amount of deposition is predicted at 10% ambient RH on the more distal late stages within the NGI than at 99% RH.
Investigating Drug Loading on Stainless‐Steel Mesh Filters used for Estimating Regional Lung Deposition
Alexander Huang1, Scott Tavernini1, Dino J. Farina2, Warren H. Finlay1, Andrew R. Martin1
1Aerosol Research Lab of Alberta, University of Alberta, 116 St & 85 Ave, Edmonton, T6G 2R3, Canada
2Proveris Scientific Corporation, 2 Cabot Rd, Hudson, MA 01749, United States
Summary: Our group has recently developed a filter‐based apparatus (FBA) for in vitro estimation of regional lung deposition for dry powder and pressurized metered dose inhalers. The FBA utilizes a set of specialized stainless‐steel mesh filters, positioned downstream of a mouth‐throat airway model (Alberta Idealized Throat, AIT), to collect and separate the tracheobronchial fraction of the lung dose. The work presented here evaluated potential loading effects of repeated actuations of inhalers through the apparatus on the filtration efficiency of the mesh filters. Varying doses of a fluticasone propionate dry powder inhaler (DPI) were actuated through the in vitro assembly while using realistic breathing profiles. Scanning electron microscopy (SEM) provided a qualitative assessment of active pharmaceutical ingredient (API) and carrier particle deposition onto the filters with increased loading, while ultraviolet (UV) spectroscopy provided a quantitative analysis of the amount of API collected on the filters. The mass of API collected on the filters, normalized by number of actuations, was unchanged between 2, 5, and 7 repeated actuations. This study therefore demonstrated the robustness of the FBA and its potential to contribute to in vitro evaluation of orally inhaled drug products (OIDPs).
Application of HPLC‐Mass Spectrometry (HPLC‐MS) in Rapid Inhaler Testing
Lei Mao, Joshua Hicks, Ryan Stack, Patricia Pinheiro, Francois Billard
Recipharm Laboratories Inc, 511 Davis Drive, Morrisville, NC, 27560, USA
Summary: Mass spectrometry (MS) detectors present a powerful option compared to UV detectors when attached to an HPLC instruments. MS‐based methods were developed at Recipharm and applied to the rapid analysis of active pharmaceutical ingredient (API) in sample solutions generated during inhaler testing. One method was developed for simultaneous analysis of a formoterol fumarate dihydrate and beclomethasone dipropionate mixture. The developed method was specific, linear, and demonstrated injection accuracy and precision with a run time of only 2.5 minutes. Application of rapid analysis with an MS detector was done comparatively from product sample solutions containing salbutamol sulphate (SS) generated during measurement of aerodynamic particle size distribution (APSD). The samples were collected from HFO1234ze pressurised Metered Dose Inhalers (pMDI). The collections were analysed using both an HPLC‐UV and an HPLC‐MS method. Assay results were equivalent between the two detectors and there was an overall 20% improvement in the run time and a 50‐fold improvement in sensitivity when using the MS detector.
A new paediatric airway model for in silico evaluation of regional aerosol deposition
Kineshta Pillay1, Warren H. Finlay1 & Andrew R. Martin1
1Aerosol Research Lab of Alberta, University of Alberta, 116 St&85 Ave, Edmonton, T6G 2R3, Canada
Summary: While there has been significant investigation into regional deposition of inhaled drugs in the airways of adults, less is known about where aerosols deposit in children's lungs. As there are challenges related to performing in vivo deposition measurements using radiolabelled particles in children, other methodologies are needed. To this end, computational models were developed to represent the lungs of girls and boys aged 8, 10 and 12 years. The models were symmetric and followed a single‐path numerical approach to calculate aerosol deposition in the airways. Total deposition efficiency calculated by these models was validated against existing in vivo data for children performing controlled breathing patterns and showed agreement for particles ranging from 1 ‐ 3 μm. Regional deposition estimates were further provided for particle deposition in children performing age‐adjusted tidal breathing manoeuvres. Differences in deposition between boys and girls were minimal, while age showed a moderate influence. Extrathoracic deposition increased with particle size, while tracheobronchial and alveolar deposition increased initially before peaking between 3 – 5 microns and decreasing thereafter. Comparison against existing regional deposition data from adults showed similar deposition patterns in the central and peripheral airways of adults and children, with lower deposition in these regions in children. It is anticipated that improved understanding of regional deposition in paediatric airways will assist in optimizing regional drug delivery to children in future work.
Solid state degradation of proteins in complex drug product formulation matrices
Gerard Masdeu1, Alisija Baliute1, Aida Edvardsson1, Jennifer Wright1 and Tatiana Starciuc1*
Summary: The scientific understanding and characterisation of pharmaceutical drugs are crucial for their success in development, especially nowadays with the emerging biologic ingredients in the inhaled drug market. The degradation pathways of protein surrounded by a solid matrix of excipients (e.g.,sugars) is complex and needs a multi‐angle characterization approach to be fully understood. Numerous publications describe the stabilisation mechanisms of proteins in the solid state, explaining how physical and chemical degradation occurs. However, there is no clear guidance on using and applying this knowledge to stress testing studies required by regulatory authorities. Herein, we suggest a novel approach to design the stress testing studies of a solid protein‐based formulation, i.e. using the glass transition temperature (Tg) of the solid matrix as incubation temperature. It is well known that the global mobility of the solid matrix, below Tg, is “frozen,” while local mobility is still possible. This will consequently reduce the protein degradation rate, due to its suggested potential link to the surrounding matrix mobility. Thus, samples in this study were incubated at temperatures above Tg, where both local and global mobility are possible. As a result, rapid degradation was achieved, evaluated by different analytical techniques in both solid and liquid state, observing secondary structure changes or higher aggregation propensity, among others. As a way forward, we suggest using the approach described in this work and challenge the scientific community to accept it.
Flow visualisations of the spray from a novel soft mist inhaler using planar laser illumination
Vishal Chaugule1, Wietze Nijdam2, Bernhard Muellinger3, Varsha Komalla4, Imco Sibum3, Hui Xin Ong4,5, Daniela Traini4,5, Nicolas A. Buchmann3, & Julio Soria1
1Laboratory for Turbulence Research in Aerospace and Combustion (LTRAC), Department of Mechanical and Aerospace Engineering, Monash University, Clayton Campus, Melbourne, VIC 3800, Australia
2Medspray B.V., Enschede, The Netherlands
3Resyca B.V., Enschede, The Netherlands
4Respiratory Technology, Woolcock Institute of Medical Research, Sydney, NSW 2037, Australia
5Macquarie Medical School, Faculty of Medicine, Health and Human Sciences, Macquarie University, NSW 2109, Australia
Summary: Soft mist inhalers are next‐generation inhaled drug delivery devices that use mechanical energy to convert a liquid formulation into a fine mist suitable for inhalation. Some soft mist inhalers create an inherently unstable liquid jet that spontaneously undergoes breakup to produce a low‐momentum aerosol cloud suitable for lung delivery. The PulmosprayTM is one such soft mist inhaler and the characteristics of the spray from its mouthpiece were qualitatively evaluated in this study using the planar laser illumination technique. The working liquid used was water for injection. The mouthpiece was provided with a steady liquid flow rate of 0.37 mL/min and steady airflow rates of 15, 30, 45 and 60 SLPM. An ensemble of single‐exposure images of the spray droplets illuminated by the laser sheet were recorded at 100 Hz using a high‐speed camera and are processed to obtain the mean spray image, for all the examined airflow rates. The macroscopic spray characteristics determined are the mean spray pattern, spray width and spray angle. Cross‐correlation analysis of single exposure images acquired at 8000 Hz was performed to determine the mean spray centreline velocity. The instantaneous spray snapshots depicted emergence of a turbulent spray for all examined airflow rates. A narrower spray was generated with increasing airflow rates, but the spray spread remained similar for the examined airflow rates greater than 30 L/min. There was a monotonic decrease in the spray centreline mean axial velocity with increasing downstream distance from the mouthpiece for the examined airflow rates.
Development and Optimisation of a Particle Sizing Methodology for Early Screening of DPI Carrier Based Formulations using Laser Diffraction.
James McNeill1, Jim Clay1, Olivia Brittain1
1Kindeva Drug Delivery Ltd Loughborough, Leicestershire, LE11 5RB, UK
Summary: Aerodynamic Particle Size Distribution (APSD) is recognised as a Critical Quality Attribute (CQA) in the in vitro characterisation of Orally Inhaled and Nasal Drug Products (OINDPs). The established instrumentation for measuring APSD of inhaled products is the multistage cascade impactor (CI). Use of laser diffraction for Particle Size Distribution (PSD) measurement offers an efficient and simplified alternative approach for early screening of Dry Powder Inhaler (DPI) formulations; however, the lack of component specificity may be a drawback. It is therefore essential to have a suitable separation strategy. In this study, a Next Generation Impactor (NGI), DPI preseparator and throat were interfaced with a Sympatec HELOS with INHALER adapter. This test configuration enabled a comparison of mean diameter (D50) from laser diffraction PSD data against mass median aerodynamic diameter (MMAD) data generated by NGI. The correlation between D50 and MMAD was improved when the standard DPI preseparator insert was coated with 1% w/v Brij® 35 in 20/80 %v/v ethanol/glycerol solution, and then further enhanced by replacing the standard insert with a 30 L/min, 5 μm cut‐off insert. Further development showed that using two stacked preseparators in series, each containing a coated 30 L/min, 5 μm cut‐off insert, generated the closest correlation between D50 and MMAD. This novel methodology could enable efficient early screening of formulations in DPI product development.
Investigating effects of rotation speed and flow rate on powder evacuation from a capsule for dry powder inhalers
1School of Aerospace, Mechanical and Mechatronic Engineering. The University of Sydney, NSW 2006, Australia
2School of Engineering, IIT Mandi, HP 175075, India 3School of Pharmacy, the University of Sydney, NSW 2006, Australia 4School of Material Science and Engineering, UNSW Sydney, NSW 2052, Australia
Summary: Capsule‐based dry powder inhalers (DPIs) have long been commercially available, but their performance remains largely unpredictable despite the prevalence of such devices. This is due, in part, to a lack of understanding of how different device designs affect powder dispersion and evacuation. One common design configuration is a spinning capsule that utilises airflow to rotate a capsule while collisions with the wall aid in dispersion. This study aimed to better understand these mechanisms by decoupling rotation speed from the flow rate and investigating their effect on powder evacuation from the capsule at various flow rates and rotation speed conditions. For this study, two flow rates, 30SLPM (inlet: 16.67 m/s) and 60SLPM (inlet: 33.33 m/s) and two rotation speed settings of 1500 RPM (Low case) and 3750 RPM (High case) were chosen. High‐speed backlit imaging further provided qualitative information about evacuation behaviour.
At 60 SLPM the rotation speed had no significant effect on the overall powder evacuation during the three second run time. Furthermore, holding the capsule stationary and in line with the flow at this speed also did not affect the amount of powder evacuated. Therefore, at this flow rate and for the formulations tested evacuation was independent of rotation speed. When tested at 30 SLPM, the lower rotation speed setting was found to improve evacuation significantly compared to the high rotation speed case. Further rotational speed cases and formulations should be analysed in combination with high‐speed microscopic imaging to gain a better understanding of dispersion behaviour.
Comparing protein stabilisation effects of inositol to mannitol and trehalose in inhalable particles prepared by nano spray‐drying
Michael Y.T. Chow1 & Jenny K.W. Lam1
1UCL School of Pharmacy, University College London, 29‐39 Brunswick Square, London WC1N 1AX, United Kingdom
Summary: Pulmonary delivery of proteins offers enormous clinical potential, and it is desirable to formulate proteins into dry powders for enhanced storage stability. In this study, the potential of inositol as a stabilising agent was compared with mannitol and trehalose. Bovine serum albumin (BSA), as the model protein, was spray dried alone or with mannitol, (myo‐) inositol or trehalose at equal mass ratio, using a nano spray dryer equipped with a mesh nebuliser. All particles exhibited a size distribution suitable for inhalation, with BSA‐only particles having a median diameter of 3.84 μm. This reduced to 3.21‐3.36 μm after incorporating stabilising agents. Contrast to pure BSA particles that showed a smooth spherical morphology, the spherical particles containing inositol or trehalose exhibited wrinkled surfaces, a desirable feature to promote deagglomeration. Inositol also resulted in a less residual moisture content (3.3%) than trehalose (5.0%) or pure BSA (5.4%), yet higher than that with mannitol (2.4%). While gel electrophoresis did not reveal detectable protein degradation across all formulations, size exclusion chromatography showed a reduction in monomer content of 4.8% in the absence of stabilisers. With inositol or trehalose, the reduction in monomer content decreased to 1.3% and 1.2%, respectively, implying protein stabilisation. To conclude, protein particles with an inhalable particle size distribution have been successfully prepared by nano‐spray drying. Inositol showed comparable protein stabilising capacities to trehalose with less residual solvent, suggesting it as a promising excipient in the formulation of protein powders for pulmonary delivery.
QbD Assisted Development of Inhalable Spray‐Dried Procubosomal Powder Intended to be used for Lung Delivery
Akash Deep & Vikas Rana
Department of Pharmaceutical Sciences and Drug Research, Punjabi University, Patiala (Punjab), India
Summary: The primary objective of study was to optimize the parameters involved in development of inhalable spray‐dried procubosomal powder (SDPP) using the spray drying process, with an aim to improved aerosolization performance and yield of formulation. The development of gefitinib (GFT) loaded‐SDPP began with establishment of a quality target product profile and a thorough risk assessment by the Ishikawa diagram that identifies ten high‐risk parameters to guide formulation development. Subsequently, the Taguchi design was employed to systematically screen different formulation parameters (glyceryl monooleate: poloxamer P407, polyethylene glycol 200 content, stirring temperature, stirring speed, stirring duration, mannitol: leucine) and process parameters (inlet temperature, outlet temperature, aspirator flow rate) along with equipment configuration parameter (cyclone design). These parameters were assessed based on their individual effects on critical quality attributes, i.e., yield (%), device removal efficiency (DRE, %), aerosolization efficiency (AE, %), and fine particle fraction (FPFTSI, %) of GFT‐SDPP. The results revealed that optimized formulation achieved a satisfactory aerosolization performance, i.e., DRE of 94.2 ± 5.6%, AE of 52.2 ± 1.84%, FPFTSI of 63.8 ± 3.63% and yield of 32.5 ± 2.82%. This optimized formulation exhibited a mean median aerodynamic diameter (MMAD) of 3.29 ± 0.53 μm, enhanced FPFAC (60.83 ± 2.19%) and entrapment efficiency (96.2 ± 3.98%). SEM analysis indicated smooth spherical particles with somewhat wrinkled surface. The results of in‐vitro dissolution studies indicated the sustained release of GFT from SDPP with enhanced dissolution. In conclusion, this study successfully developed an inhalable SDPP formulation, demonstrating promising potential as targeted drug delivery system.
KuDa‐tob: an Inhaled tobramycin formulation to improve treatment of Pseudomonas aeruginosa chronic lung infections for cystic fibrosis patients
Julie Movellan1, Núria Blanco‐Cabra2,3, José María Marimón4, Eduard Torrents2,3, Damien Dupin1, Hans‐Jürgen Grande1,5 & Iraida Loinaz1,6
1CIDETEC, Basque Research and Technology Alliance (BRTA), Parque Científico y Tecnológico de Gipuzkoa, 20014 Donostia‐San Sebastián, Spain
2Bacterial Infections: Antimicrobial Therapies group, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), Barcelona, Spain.
3Bacterial Infections: Antimicrobial Therapies group, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), Barcelona, Spain.
4Microbiology Department, Donostia University Hospital ‐ Biodonostia Health Research Institute Donostia‐San Sebastian, Spain.
5University of the Basque Country (UPV/EHU), Advanced Polymers and Materials: Physics, Chemistry and Technology Department, Donostia‐San Sebastian, Spain
6Kusudama Therapeutics, Parque Científico y Tecnológico de Gipuzkoa, 20014 Donostia‐San Sebastián, Spain
Summary: Cystic fibrosis (CF) is characterized by abnormal mucus hydration due to a defective CFTR protein, leading to the production of difficult‐to‐clear mucus. This results in airflow obstruction, recurrent infections, and respiratory complications. Tobramycin is the first line of antibiotic treatment against chronic infections of Pseudomonas aeruginosa (PA). Administering the complete daily dose of tobramycin in a single inhalation per day may offer advantages over dividing the dose into two sessions, potentially reducing adaptive resistance, and improving treatment adherence and patient quality of life.
KuDa‐tob, a nanoformulation of tobramycin and dextran single chain nanoparticles platform (KuDa) as an excipient, was specifically developed to enhance the bioavailability of the antibiotic. The neutralization of the positive charges of the drug facilitates its diffusion through the mucus and biofilm, reaching the bacteria. In vitro studies have shown that KuDa maintains the antibiotic's efficacy and promotes its penetration into the biofilm.
Freeze‐dried KuDa‐tob can be reconstituted for nebulization and redispersed easily at high concentrations without affecting the droplets diameter. Nebulization of KuDa‐tob to mice showed similar efficacy between KuDa‐tob administered once daily double dose compared to tobramycin in solution administered twice a day single dose. These findings confirmed the potential of KuDa‐tob as a once‐daily treatment for chronic infections caused by PA for CF patients. Further research and evaluation are currently under progress to confirm its clinical effectiveness and safety.
Digital tool development ‐ Modelling Recrystallization of Amorphous Lactose.
1AstraZeneca Gothenburg, Pepparedsleden1, 431 84, Mölndal, Sweden
Summary: The need for more digital tools to support development and manufacturing has increased a lot in all types of industries in the last decades, the pharma industry is no exception. In this study, a modelling tool has been developed to support the process of recrystallization of amorphous powders. This is of particular interest for inhalation due to the impact of residual amorphous on stability or aerosolization performance of products.
In this work, the crystallization of amorphous lactose was simulated and compared with isothermal microcalorimetry, TAM (Thermal Activity Monitor) experiments. The driving force for recrystallization used the difference between the glass transition temperature (TG) of the amorphous lactose and the operating process temperature, T. The Gordon & Taylor equation is used to describe the composition dependence of the transition temperature. The Guggenheim–Anderson–de Boer (GAB) equations with parameters fitted for the different states of lactose are utilized in this study to estimate saturated moisture content at different humidity levels. The crystallization kinetics used are the ones proposed by Bronlund [1], namely Avrami‐Bronlund.
Development of inhalable dry powders combining replication and protease inhibitors of SARS‐CoV‐2
Tushar Saha1, Shubhra Sinha2,3, Rhodri Harfoot2, Miguel E. Quiñones‐Mateu2,4 & Shyamal C. Das1
1School of Pharmacy, University of Otago, Dunedin, 9016, New Zealand
2Department of Microbiology and Immunology, School of Biomedical Sciences, University of Otago, Dunedin, 9016, New Zealand
3Current address: Department of Physiology, University of Otago, Dunedin, New Zealand;
4Current address: Department of Pathology and Laboratory Medicine, Western University, London, ON N6A 3K7, Canada
Summary: Respiratory drug delivery is considered the most logical approach to treating respiratory tract infections, including severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2). Delivering antiviral agents directly to the respiratory tract can ensure effective drug concentrations at the infection site with less exposure to the systemic circulation causing less side effects. The combination of replication (remdesivir) and protease (disulfiram/ebselen) inhibitors of SARS‐CoV‐2 can block virus production synergistically in vitro. This data gives an important insight that, suitable drug combinations will not only offer better antiviral effects but also restrain the chance of upcoming drug resistance against SARS‐CoV‐2, as it is harder to develop drug resistance against multiple drugs than a single drug. Considering this, inhalable dry powders containing remdesivir, in combination with disulfiram or ebselen, were developed utilizing spray‐drying technology. The inhalable sized (1.6 − 2.9 μm) combinational dry powders exhibited spherical shapes and were crystalline in nature. The emitted dose and fine particle fraction of the prepared combinational dry powders varied between 80–88% and 55–58% respectively. Furthermore, the dry powders were non‐toxic to respiratory cell lines (CC50> 40 μM) and retained antiviral properties with EC50 values ranging from 4.43 – 8.04 μM.
Identifying determinants of capsule emptying
Mark Sanders1 & Darragh Murnane1
1University of Hertfordshire, College Lane, Hatfield, AL10 9AB, UK
Summary: Single dose capsule inhalers are a popular dry powder delivery system, and a number of advantages for the patient and manufacturer; there is a positive reinforcement rattle sound that implies delivery. The ERS/ISAM Task Force report (1) explains that the dose from a capsule DPI is emitted slower than that of a reservoir or blister DPI and that therefore inhalation volume is an important parameter for patients using capsule inhalers. A UH project has investigated what other parameters may be relevant determinants of efficient capsule emptying and found that duration of inhalation, flow rate of inhalation, power generated and device resistance all make a contribution to capsule emptying.
Impact of salts on spray dried inhaled dual site targeting mannitol powder for radiolabelling procedure
Han Cong Seow1,2, Michael Y.T. Chow1, Zhu Zilan1, Mridul Majumder3 & Jenny K.W. Lam1,2
1Department of Pharmaceutics, UCL School of Pharmacy, University College London, 29‐39 Brunswick Square, London, WC1N 1AX, UK
2Department of Pharmacology and Pharmacy, LKS Faculty of Medicine, The University of Hong Kong, 21 Sassoon Road, Pokfulam, Hong Kong SAR
3M2M Pharmaceuticals Ltd., Unit 125 (Gr Fl) Wharfedale Road Winnersh Triangle RG41 5RB, Berkshire, UK
Summary: Dual site targeting approach is an innovative aerosol drug delivery strategy to target both upper and lower respiratory tracts by intranasal administration. The idea is to produce powders of distinctly large (>10 μm) and small (<5 μm) sizes for nasal and lung deposition, respectively, and by manipulating the powder mixing ratio, their deposition profile can be controlled. To validate this concept, we aim to produce radiolabelled powders for a clinical deposition study. During radiolabelling, substantial amount of salts are added to stabilise the radioactivity. As high salt content may affect the spray drying process and the physicochemical properties of the powder, our main objective was to investigate the effect of salts on the powder produced by co‐spray drying mannitol with salts required for radiolabelling. It was found that the additional salt content decreased the production yield, increased the residual moisture and particle size of the powders. By blending the powders of different size, the particle size distribution varied linearly with the mixing ratio. This linear trend could still be observed in the powders containing 10% w/v salt. Overall, a dual site targeting powder formulation of mannitol for radiolabelling was developed for subsequent use in clinical deposition study to validate the dual targeting formulation approach as a promising strategy for delivering therapeutic agents to the nasal cavity and the lungs.
A new proposal for the development of sustainable generic DPI products based on respirable dose matching rather than being an exact copy of the reference
1Food and Drug Department, University of Parma, Italy
2Plastiape, a Berry Global company, Osnago, Italy
Summary: In contrast to the current strategy, which encourages generic manufactures to create “low performing” formulations in order to match the aerodynamic characteristics of out‐of‐date reference product, this work provides evidence in favour of an unconventional sustainable approach to create generics. Thanks to the advances in device and formulation, greater drug emitted, and respirable doses can be achieved with the possibility to load a lower nominal dose hence reducing the drug mass employed and the impact related to its synthesis and disposal.
The purpose of this project was to assess the reduction of the nominal dose of two types of exploratory generic formulations produced with two different technologies (by mixing in Turbula mixer and by spray drying process), coupled with RS01 inhaler as copies of the reference products. The reference products were Spiriva® Handihaler®, containing tiotropium bromide (TiB), and Serevent® Diskus®, containing salmeterol xinafoate (SX).
From the data obtained, it is possible to reduce the drug nominal dose of 30% and 50% for the TiB‐InhaLac150_RS01 and TiB‐Spray Dried (SD) with Leucine_RS01 formulations, compared to the reference product; rather, considering the SX‐InhaLac150_RS01 and SX‐SD with Leucine_RS01 formulations, it is possible to have a drug nominal dose reduction of 25% and 33% respectively.
This work represents a provocation to push us to think in a more “modern” and sustainable way but certainly it should be agreed by the regulatory authorities and the nominal dose reduction would have to be confirmed in vitro along with a clinical study showing efficacy and PK profile.
Minimal amounts of sodium chloride combined with leucine improve aerosol performance of spray‐dried vardenafil powders for inhalation
Qingzhen Zhang1,5, Zheng Wang1,2, Philip Hall1,3, Yan Zhao4, &Bin Dong4
1Department of Chemical and Environmental Engineering, University of Nottingham Ningbo China, Ningbo 315100, People's Republic of China
2Key Laboratory for Carbonaceous Wastes Processing and Process Intensification Research of Zhejiang Province, University of Nottingham Ningbo China, Ningbo 315100, People's Republic of China
3Nottingham Ningbo China Beacons of Excellence Research and Innovation Institute, Ningbo 315100, PR China
4Pharmaceutical Engineering, China Pharmaceutical University, Nanjing,211198 P.R. China
5Suzhou Inhal Pharma Co., Ltd, Suzhou, Jiangsu, 215000, China
Summary: This study aimed to develop highly dispersible vardenafil dry powder with good lung deposition by spray drying aqueous solution of vardenafil and leucine with 0%, 2.5%, 5% and 10% of sodium chloride (by weight) based on vardenafil concentration. The effect of concomitant use of leucine and minimal amount of sodium chloride on powder morphology and aerosolization performance were investigated under various inhalation air flow rates and powder filling amounts. The dispersibility in different capsule filling amount and air flow rate presented low variation with 2.5% sodium chloride formulation. This optimized dry powder formulation may potentially improve the onset of action, reliability, and safety for clinical application of vardenafil.
In Vitro and in Vivo Assessment of Nanoporous Amorphous Particles as a Potential Excipient for Inhaled Drug Products
Georgia A. Pilkington1,2, Bjarne Falk1, Konstantinos Paraskevas1, Ewa Selg3 and Per Gerde3 & Gary Pitcairn1,4
1Nanologica AB (publ), Södertälje, Sweden
2Division of Surface and Corrosion Science, KTH Royal Institute of Technology, Stockholm, Sweden
3Inhalation Sciences, Huddinge, Sweden
4GRP Consulting AB, Gothenburg, Sweden
Summary: Nanoporous amorphous silica particles (NAP) are being developed as potential excipient for creating dry powder formulations. As part of the development process, it was deemed necessary to understand how long the particles would remain in the lung as this could have potential safety implications for chronic use. In the present study, it was shown that the surface area of the particles influenced the how quickly the particles dissolved in vitro, and that faster in vitro dissolution resulted in faster particle clearance from the lungs of rats. The NAP manufacturing process was developed to successfully produce a particle with a high surface area (NAP‐543). Based on the in vitro/in vivo relationship observed, the expectation is that NAP‐543 would be cleared from lung within 6 hours and thus could potentially be suitable as an excipient for formulating drug products intended for once daily dosing. Further work is required to scale up the manufacturing process and, critically, demonstrate the safety of this novel excipient in preclinical studies.
Regional Deposition of Spray Dried Trehalose Powders in the Alberta Idealized Nasal Inlet
Kelvin Duong1, Maximilian Aisenstat1, John Z. Chen3, Brynn Murphy1, Scott Tavernini1, Kyle Kaufman1 Jing Zheng2, Randy Whittal2, Alana Gerhardt3, Christopher Fox3, Warren H. Finlay1, Reinhard Vehring1, & Andrew R. Martin1
1Department of Mechanical Engineering, University of Alberta, Edmonton, AB, T6G 2R3, CA
2Mass Spectrometry Facility, University of Alberta, Edmonton, AB, T6G 2R3, CA
3Access to Advanced Health Institute (AAHI), Seattle, WA, 98102, US
Summary: The present study compares the in vitro regional deposition of spray dried trehalose in the Alberta Idealized Nasal Inlet (AINI) using nominally small (predicted MMAD ∼5.5 μm) versus large (predicted MMAD ∼18 μm) primary particle sizes. An active, single dose device was used with automatic actuation to deliver the trehalose powders for both particle sizes at three different loaded doses, 10 mg, 20 mg, and 40 mg. A test flow rate of 7.5 L/min was maintained through the AINI. The interior of the AINI was coated prior to actuation to mitigate particle bounce. Deposition of trehalose in the four regions of the AINI (vestibule, turbinates, olfactory, and nasopharynx), pre‐separator, and filter (representing in vitro lung deposition) was determined for each combination of particle size and loaded dose using liquid chromatography coupled with mass spectrometry by multiple reaction monitoring.
In all cases, a high fraction of trehalose powder penetrated past the vestibule into the turbinates and nasopharynx. For all three doses of the small particle size powder, a small yet non‐negligible fraction of powder reached the filter, representing lung exposure. In the case of large particles, the fraction depositing on the filter was negligible, less than 0.7% of total recovered dose.
In summary, penetration of trehalose powder through the AINI was eliminated using larger primary particles, while high turbinate deposition was maintained.
Use of Rheology and Mucoadhesivity to Guide the Development of Nasal Mucus Simulants
Lucy Goodacre1, Jana Janorovic1, William Ganley2, Irene Rossi2, Cecile Dreiss1 & Ben Forbes1
1King's College London, Institute of Pharmaceutical Science, 150 Stamford Street, London, SE1 9NH
2Nanopharm Ltd, an Aptar Pharma Company, Franklin House, Grange Road, Cwmbran, South Wales, NP44 3WY
Summary: Respiratory mucus models (or simulants) are surprisingly little used for the development and quality control of nasally‐delivered drug products. Local and systemic exposure following nasal administration of medicines is determined by interactions with mucus at the site of deposition. However, mucus is seldom incorporated into experimental models used to evaluate product performance, e.g. deposition, absorption or dissolution studies. To guide the development of mucus simulants for use in such studies, our aim was to establish a suite of complementary analytical methods to characterise the bulk rheology, micro‐rheology and mucoadhesivity.
Putative artificial, synthetic and native mucus simulants were investigated, with an initial focus on an 15% w/w Pluronic® F127 gel as an example. Bulk rheology was measured using an ARES strain‐controlled rheometer and indicated gel‐like rheological properties (G’ 4.27 ± 1.48 kPa and G” 1.60 ± 0.21 kPa). Micro‐rheology was characterised by particle tracking using a Nikon Eclipse Ts2R microscope (diffusion coefficient 0.5814 μm2/s with 95% confidence interval 0.5763 ‐ 0.5867 μm2/s) and mucoadhesivity was measured using an Instron® 5500A series (median AUC 0.1757 N*mm; range 0.1299 ‐ 0.2476 N*mm). Collectively, these techniques provide a means of assessing relevant functional properties that can be used to characterise and guide the development of mucus simulants.
High speed video analysis for the characterisation of a jet for targeted nasal drug delivery
Lenaic Guimard1, Benjamin Ogier2 & Alain Regard1
1Nemera La Verpillière, 63‐65 Avenue Tony Garnier, Lyon, 69007 – Lyon, France
2Abylsen for Nemera La Verpillière, 63‐65 Avenue Tony Garnier, Lyon, 69007 – Lyon, France
Summary: Most of commercial nasal combination products are based on spray of liquid solutions and suspensions for local and systemic treatments. Regulatory guidelines are defining the characterisation methods of the spray required in the scope of the quality control of the combination products and in the scope of in vitro bioequivalence. Droplet size distribution, spray pattern and plume geometry are commonly measured using dedicated characterisation techniques and equipment.
With the development of treatments using the direct Nose‐to‐Brain pathway, targeting the upper nasal cavities and more specifically the olfactory cleft is key in the efficacy of such treatments. One way to target the olfactory cleft with liquid formulations or powder formulations is to use a narrow spray or jet‐like delivery.
When it comes to the characterisation of plume for a jet, the physical properties differ, and the use of spray characterisation equipment becomes not relevant. Nemera® has developed an equipment (hardware and software) to analyse the physical characteristics of a jet to support a nasal jet‐based device development.
The hardware is based on a high‐speed camera and an automated actuator precisely positioned on an optical bench. The video analysis software allows a characterisation of jet cone angle, deviation of this cone from device axis, width at given distances and jet front speed.
The hardware and software developed provide relevant data to support a jet‐based device development. Additional development and validation work will be performed to make this tool match the pharmaceutical industry requirements.
Targeted drug delivery to the olfactory cleft: In vitro deposition study using prototype jet nozzles.
Alain Regard1, Benjamin Ogier2 & Laura Berard1
1Nemera La Verpillière, 63‐65 Avenue Tony Garnier, Lyon, 69007 – Lyon, France
2Abylsen for Nemera La Verpillière, 63‐65 Avenue Tony Garnier, Lyon, 69007 – Lyon, France
Summary: Nasal treatments using nose to brain pathway are a promising way of addressing CNS diseases. One challenge of the nose to brain treatments remains the targeted administration of the drug formulation to the olfactory cleft. Nemera is exploring the ability of a narrow and straight jet to deliver drug to the olfactory cleft.
Seven prototype devices have been designed and manufactured with the aim to produce a highly stable jet. In vitro deposition tests have been performed with these prototype devices on two different nasal casts with a liquid placebo formulation. Qualitative data and quantitative data related to the deposition in the nasal cavity have been produced.
The results from these two casts are significantly different: high level of deposition in the olfactory area (65 to 88 %) and extremely low variability have been observed with the Athena nasal cast; with the FAN II nasal cast, the deposition levels are significantly lower with a high difference in the mean values and an important variability.
These contradictory results open a field of investigations regarding the effect of the potential gaps between the two experimental conditions such as different anatomies, differences in the method of extraction of the formulation from the casts and quality of the surface of the nasal casts.
This study shows how critical it is to confront different nasal cast models to assess the deposition performance of nasal devices.
In Vitro Characterisation of Octreotide Microspheres Formulated for Nose to Brain Delivery
1Nanopharm Ltd, an Aptar Pharma Company, Grange Road, Cwmbran, NP44 3WY, United Kingdom
2Biodexa Pharmaceuticals, Caspian Way, Cardiff, CF10 4DQ, United Kingdom
Summary: Octreotide is a metabolically stable analogue of a somatostatin, a potent inhibitor of secretory processes, and has promising results against the neurological condition idiopathic intracranial hypertension when delivered subcutaneously. Delivery of octreotide intranasally to the brain by bypassing the blood brain barrier and targeting the central nervous system through the olfactory region of the nasal cavity can increase its efficacy. Biodexa Pharmaceuticals have developed Q‐SpheraTM octreotide loaded poly(lactic‐co‐glycolic acid) (PLGA) microspheres that could be promising for nose‐to‐brain delivery by increasing residence time, controlling the release of the encapsulated drug and reducing systemic side effects. The aim of this study was to formulate and assess the suitability of delivering the Q‐SpheraTM for nose‐to‐brain with Aptar VP7 coupled with 232‐NE actuator. A suitable formulation for nasal delivery was prepared as indicated by the pH value of 5.05 and osmolality value of 491 mOsm/kg. No effect of shear stress on the microspheres was observed with the same PSD obtained before and after spraying. The microsphere formulation displayed a large median droplet size (134 μm) due to its higher viscosity (3.7 cps) in comparison to an octreotide nasal solution (66 μm). Despite the larger droplet size, the in vitro nasal cast deposition study indicated a significantly higher deposition in the nasal region of interest, the olfactory area, for the microspheres' nasal formulation, due to the narrower plume emitted. These results indicate that the microspheres can be successfully formulated for nasal administration showing spray characteristics that are suitable for nose‐to‐brain delivery.
An investigation of actuation forces and performance of a novel, user‐independent nasal spray, based on user‐device interaction
Summary: Single dose nasal spray device users fall into a wide demographic, with varying abilities, experience and requirements. These differences, combined with a general need for accurate and reliable drug delivery, highlight the importance of universally appropriate user‐device interaction and the benefits of a platform device. The main purpose of the work described in this paper was to assess Recipharm's proprietary, novel unit dose (NasaDose™) nasal spray device capability with respect to consistent and appropriate actuation forces at two different potential drug formulation viscosities. For this device, all of the delivery force is spring‐driven, minimising the effort required, and force experienced, by the end user. As the device has a trigger‐based mechanism, the design must ensure conflicting requirements of user ability to actuate and robustness upon drop are met. User actuation capability was assessed by measuring the forces that they could comfortably apply to a model nasal spray, as part of a formative usability study. The data revealed a broad “comfortable actuation” force range of 11.1‐52.8 N (mean minimum and maximum for all study participants; relative standard deviation (RSD) 12.3% and 6.8%, respectively). Actuation force and metered shot weight (MSW) were measured after drop tests on two sets of device variants. No actuation of devices occurred when dropped, and there was negligible variation in MSW and slight variation in actuation force after dropping samples. This supports the hypothesis that Recipharm's proprietary device performs repeatably and reliably, and operates within an identified range of comfortably achievable user actuation forces.
Evaluation of the Performance of a Blister Unit Dose Nasal Powder Device
1Aptar Pharma, Route des Falaises, Le Vaudreuil, 27100, France
2Nanopharm Ltd, An Aptar Pharma Company, Grange Road, Cwmbran, NP44 3WY, United Kingdom
3TEKH Consulting Ltd, Queen Street, Deal, CT13 6EY, United Kingdom
Summary: A novel single dose nasal powder (UDSp) passive device incorporating a blister dosing chamber was developed. The assessment of the performance of the device was carried out by using a model spray dried formulation, which was filled semi‐automatically by drum technology using the Omnidose Drum TT. The device performance was tested applying three flow rates: 15, 30 and 45 L/min. Shot weight and emitted dose uniformity reported a very high percentage of powder mass and active ingredient emitted, 96% and 99% of the loaded powder, respectively, independently of the flow rate applied. In addition, the emitted particle size distribution results obtained showed a Dv50 of about 40 μm, suitable for nasal delivery with less than 2% of particles <10 μm, once again with no correlation with the flow rate applied. This was also reflected on the aerodynamic particle size distribution results, which reported 95% of the active deposited in the mouthpiece, induction port and stage 1. Generally, this novel UDSp blister device proved to be capable of delivering a consistent dose and a suitable particle size for nasal delivery with no dependency on flow rate employed to actuate the device. Therefore, this relatively simple device can potentially be easily employed for different applications, such as nasal vaccination. Next step in the device characterisation will be focused in determining its performance in combination with powders with different physical characteristics, exploring different filling processes and fill weights.
Investigation of the Performance of a Novel Unidose Powder Blister Device for Nasal Delivery
Cristina Rey‐Blanes1, Reanne Beaird1, Nathalie Hauchard2, Angelo Consalvo3, Thomas Daggs3, Paul Shields3, Gonçalo Farias2 & Irene Rossi1
1Nanopharm Ltd, An Aptar Pharma Company, Grange Road, Cwmbran, NP44 3WY, United Kingdom
2Aptar Pharma SAS, Route des Falaises, Le Vaudreuil, 27100, France
3Enteris BioPharma Inc, Fulton St, Boonton, New Jersey, 07005, United States
Summary: The performance of a novel unidose blister device (UDSp blister) for nasal powders was assessed. Three different powders were used: Lactose LH400, a spray dried Hypromellose/Mannitol formulation and a lyophilised/Resonant Acoustic Mixing formulation composed of N‐dodecylphosphocholine/β‐Cylodextrin (DPC/β‐CD). All powders had a distinct particle size (PSD) and rheological properties, which determined the fill weight obtained by vacuum drum filling. The assessment of powder rheology showed the DPC/β‐CD powder to be the most cohesive, possibly due to its composition and manufacturing process. This was reflected in the emitted mass, where the lyophilised formulation presented the lowest fraction emitted (<10% at 15 L/min) that increased (<50%) at 30 L/min. LH400 was impacted by fill weight, especially at 20 mg, where the blister space was not enough to allow for a correct release of the dose. The spray dried formulation presented the best results when combined with the UDSp blister device, especially for the higher fill weights, with no impact of the flow rate applied on the emitted fraction. Moreover, the emitted particle size distribution of the spray dried and LH400 showed that neither the flow rate nor the fill weight had an impact on the size of the particles delivered, which also remained comparable to the bulk PSD measured. This study highlighted the importance of formulation development, excipient selection and manufacturing process for intranasal powder delivery. Next steps will include further investigation and optimisation of the powders' composition, for example by evaluating model drugs, and manufacturing process for coupling with a passive nasal powder device.
The interplay of formulation viscosity with device type and inspiratory flow provides an opportunity to target regional nasal deposition
Tsz Yan Alice Fong1, Teresa Iley2, Mark Parry2 & Ben Forbes1
1King's College London, Institute of Pharmaceutical Science, London SE1 9NH, UK
2Intertek, Melbourn, UK
Summary: This study investigated the effect of formulation viscosity in two different device types on spray characteristics and regional deposition in the nasal cavity. The influence of inspiratory airflow on nasal deposition was also investigated. Low and high viscosity formulations of a model drug, caffeine, were produced by varying the concentration of carboxymethyl cellulose. The formulation was incorporated into a conventional pressure‐swirl actuator represented by Aptar® CPS and a novel Medspray Rayleigh nozzle® with a Medspray nasal soft mist device (low viscosity formulation) or a syringe (high viscosity formulation). The generated spray characteristics were assessed by measuring plume geometry, droplet size distribution, and deposition in a nasal cast operated at two different airflows, mimicking resting conditions and a ‘sniff’. Initial results indicated that the Medspray nasal device was unable to spray the high viscosity formulation, so the actuator nozzle was used with a syringe instead. Using a standard set‐up, plume geometry could only be measured with the Aptar device, with the higher viscosity formulation producing narrower and more circular plume, the droplet size was larger compared to the lower viscosity formulation. The spray from the higher viscosity formulation resulted in a lesser turbinates deposition; greater formulation fraction was recovered from the olfactory region compared to the lower viscosity formulation. Contrarily, the Medspray nozzle produced a smaller droplet size irrespective to formulation viscosity, which, like the Aptar device, deposited predominately in the turbinates. Greater airflow simulated ‘sniffing’, which facilitated greater turbinates deposition in the distal nasal cavity.
High precision determination of amorphous lactose formed during comminution processes
Ross Blezard1, Timo Roelofs1 & Pauline H.M. Janssen1,2
2Department of Pharmaceutical Technology and Biopharmacy, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands
Summary: The presence of amorphous lactose in commercially available lactose grades used as a carrier in dry powder inhalers has been the subject of much discussion. In order to understand if the presence of amorphous lactose can impact drug performance one must first be able to measure and control amorphous lactose precisely. When formed through comminution processes such as milling or micronisation, the amorphous lactose phase is particularly unstable, making precision determination challenging. In this study an existing Differential Scanning Calorimetry (DSC) method was optimized such that the precision of the method for comminution formed lactose was improved by approximately one order of magnitude. This enables more reliable measurements and subsequently better understanding of correlations between this specific property and drug product performance.
Development of innovative controlled released dry powders for inhalation based on budesonide for a once daily administration
Le Van Bui1, Elena Menchi1, Ismaël Hennia1, Karim Amighi1 & Nathalie Wauthoz1
1Unit of Pharmaceutics and Biopharmaceutics, Department of Pharmacotherapy and Pharmaceutics, Faculty of Pharmacy, Université libre de Bruxelles (ULB), Boulevard du Triomphe CP207, B‐1050 Brussels, Belgium. *equal contribution, Le.Bui@ulb.be
Summary:Introduction. Dry powder inhalers (DPIs) in the treatment of non‐communicable chronic respiratory diseases such as asthma and chronic obstructive pulmonary disease (COPD) are one of the most widely used devices used by the patients. However, the appropriate use or choice of inhalable medications can be challenging for patients and healthcare professionals. Poor adherence due to high frequency of administration, errors in inhalation technique and/or use of DPIs may cause the loss of asthma or COPD control. In the present study, we aim to develop a dry powder for inhalation (DPI) formulation based on controlled release of budesonide (BUD) using a lipid matrix based on hydrogenated castor oil (HCO) and tocopherol polyethylene glycol 1000 succinate (TPGS) to decrease the administration frequency. Methods. BUD formulations were produced using the spray‐drying particle engineering technique. The aerodynamic properties were assessed using the Next Generation Impactor (NGI) according to the European Pharmacopeia. The release properties of BUD were assessed using a paddle apparatus with a Fast Screening Impactor cassette for dissolution. Results. The formulations showed optimal aerodynamic properties with a fine particle fraction related to the nominal dose (FPFn) comprised between 52.5% to 53.5% as well as drug releases that decreased depending on the content of HCO (20 (% w:v) or 95 (% w:v)) and TPGS (5% (w:w) of the excipients). Conclusion. Our results suggest that the content of the lipid matrix in HCO and TPGS is responsible of the extended drug release in PBS – 0.02% Tween20 (w:v) and fine particle ratio (FPF).
Transition to renewable materials: environmental impact assessment of renewable polymers for conceptual multidose dry powder inhaler
Summary: Most inhalation devices are manufactured with polymers derived from crude oil, a depleting fossil fuel the use of which emits significant greenhouse gas (GHG) emissions. To achieve sustainability goals, like NetZero targets, it is critical to divert away from fossil fuels and accelerate the transition to renewable material sources, like biopolymers.
Biopolymers can come from a range of sources, typically from chemical processing of food waste, crop waste or non‐food competing crops, all of which are from agricultural origin. In recent years there have been an increasing number of ‘drop in’ renewable polymers suitable for manufacture of inhalers, with lower GHG emissions and reduced dependency on non‐renewable energy sources. However, due to agricultural and chemical processing they can often lead to greater environmental burden in other impact categories [1], which can severely impact the biodiversity of crucial habitats.
This cradle‐to‐gate life cycle assessment (LCA) conducted on a conceptual multidose dry powder inhaler (DPI) illustrates a comparison between a fossil based and renewable polymer device. By looking at production of the device only, moving to renewable materials can lead to GHG emissions reducing by 31%, but freshwater eutrophication increasing by 75%.
The need to transition to renewable materials is great but doing so can introduce unexpected and severe environmental impact. Use of LCA is crucial to unearthing valuable insight to engage with stakeholders across the supply chain to ensure reductions in carbon footprint is maximised and impacts like eutrophication are mitigated.
Automated Process Development on the Vacuum Drum Filling of DPI Formulations
Isabel Gallego1,2, Thomas Brinz1, Bernhard Wagner1 & Martin Sommerfeld2
1Syntegon Technology GmbH, Stuttgarter Str. 130, 71332 Waiblingen, Germany
2Institute of Process Engineering, Multiphase Flow Systems, Otto‐von‐Guericke‐University Magdeburg, Hoher Weg 7b, 06120 Halle (Saale), Germany
Summary: Automated process development (APD) combines design of experiments (DoE) with automated experiment execution. This method allows to reduce the number of experiments needed to optimize the process in a fast and efficient way. The filling of capsules for dry powder inhalation (DPI) applications using vacuum drum was investigated using the APD method. Due to the low amount of powder dosage for DPI applications, accurate fill weight and content uniformity are very important. The set of machine parameters with which the capsules are to be produced must be carefully chosen according to the formulation to be filled. A DoE was performed to investigate the effect of different factors (stirrer geometry, stirrer speed, stirrer direction, powder bed height and machine speed) on mean capsule fill weight and relative standard deviation (RSD) of the fill weight. Additionally, the content uniformity of the capsules was investigated, in which the coarse particles of a mixture (d50 = 136 μm) were previously dyed with methylene blue, to be able to measure their concentration in the capsules with spectrophotometry after each run. The concentration of coarse particles, as well as the RSD of the concentration were taken as two additional responses on the DoE. The results indicate that the direction and the interaction between direction and stirrer speed have the highest effect on the responses. It was concluded that when choosing the optimal set of parameters, it is necessary to consider not only minimizing the RSD of the fill weight, but also minimizing segregation, by analysing concentration and its RSD.
The role of mechanical properties in DPI Carriers
Zeredescht Majid1, Constanze Mueller2, Ricarda Leister2 & Regina Scherließ1
1Department of Pharmaceutics and Biopharmaceutics, Kiel University, Gutenbergstrasse 76, 24118 Kiel, Germany
Summary: The aerodynamic performance of interactive blends is known to be influenced by mixing time and energy, but reasons remain poorly understood. The leading hypothesis as to explain why the fine particle fraction (FPF) decreases over increasing mixing time is based on the effect of so‐called press‐on forces. Plastic deformation of particles is seen a requirement for press‐on forces to have an effect. If this hypothesis holds true, it follows that interactive powder mixtures comprising carriers with a higher propensity for plastic deformation would exhibit a higher sensitivity to these press‐on forces compared to mixtures with carriers possessing a lower tendency for plastic deformation (brittle materials). Building upon this notion, we explored the idea by developing model carriers of different mechanical properties using Eudragit RL and varying amounts of PEG 4000 as a plasticiser. By adjusting plasticiser content, we were able to control the deformability of the carriers. To assess deformability, we fabricated films from these microspheres and determined their Young's modulus under the assumption that the tensile deformation behaved similarly to the particles experiencing compressive stress. Utilising these model carriers, we created mixtures using a high shear mixer, took samples at different intervals of mixing time, and subsequently evaluated their aerodynamic performance. Through this systematic investigation, we successfully identified a correlation between the mechanical properties of the carrier and the maximum attainable FPF, as well as the decrease in FPF with mixing time.
Investigation and Resolution of Particle Re‐entrainment in NGI Analysis of a Spray Dried Formulation Emitted from a Dry Powder Inhaler
Eric Peterson1, Cameron Kadleck1, Jordan Oliver1, Adam Schneiderhan1, Maureen Kadleck1, Karina Joyce2, Max Allsworth2, Jacob Harker3
1Lonza AG, 63045 Corporate Place, Bend, Oregon 97701, United States of America
2Owlstone Medical, 183 Cambridge Science Park, Milton Rd, Cambridge CB4 0GJ, UK
Summary: Aerodynamic Particle Size Distribution (APSD) is a key metric for pharmaceutical products intended for inhalation delivery, primarily determined using multistage cascade impactors like the Next Generation Pharmaceutical Impactor (NGI). Particle re‐entrainment can severely affect measurement of the aerosol size profile, inhibiting design and engineering of respirable particles. In this study, several industry‐standard pan coating solutions displayed high degrees of re‐entrainment during NGI testing of a spray‐dried intermediate (SDI) designed as a lung cancer diagnostic. Instead of allowing the coating solution to dry before testing, loading the pans with an aqueous solution was found to greatly reduce re‐entrainment, enabling more accurate APSD determination. In our study, the commercially available high resistance RS01 Dry Powder Inhaler (DPI) was used to aerosolize a powder engineered to have an almost entirely respirable aerodynamic particle size distribution. The liquid loading method performed well with 10 or 40mg capsule fill‐weights, while more typical impactor adhesive coatings gave significantly different results, which were also dependent on capsule fill‐weight.
High drug load and performance formulation platform for DPIs
Susana Saldanha1, Filipe Vultos2, João Pires1 & Eunice Costa1
1Inhalation and Advanced Drug Delivery, R&D, Hovione FarmaCiência SA, Lumiar, 1649‐038, Portugal
2Analytical Development, R&D, Hovione FarmaCiência SA, Lumiar, 1649‐038, Portugal
Summary: The work reports the development of an enhanced inhalable pharmaceutical formulation for high dose delivery to the lungs that comprises spray dried crystalline particles coated with a force control agent. Carrier‐free dry powder formulations for inhalation comprise a widespread array of presentations for the delivery of active ingredients to the lungs. Composite particles manufactured by spray‐drying from a solution, for high load formulations, are well established and can lead to suitable aerodynamic performance. Nevertheless, these dry powders often require more challenging storage and handling conditions to ensure their stability due to the amorphous state of the API and of stabilizing excipients. Moreover, amorphous API can result in a faster dissolution which may not be suitable for specific indications. Consequently, Spaycoat, a formulation platform for high drug load formulations comprising crystalline API and relatively low percentage of excipients (by coating the API surface) can circumvent some of these drawbacks towards more effective medicines. The results demonstrate that crystalline active ingredient particles coated with a force control agent, such as Leucine, provides a solution for delivering high drug loads to the lungs by increasing more than 4 times the fine particle fraction (FPF) from 7% to 38% when comparing with the uncoated API particles. The results show that this formulation platform can be applied in a wide range of therapeutic indications that require high drug load inhalable powders. In addition, the combination of the formulation with a high performance device such as Sunriser®, can further improve the delivered dose efficiency.
Impact of the blending process of carrier‐based formulations on DPI performance
1Instituto Superior Técnico, DEQ, Lisboa, 1049‐001, Portugal
2NOVA School of Science and Technology, DQ, Caparica, 2829‐516, Portugal
3Analytical Development, R&D, Hovione FarmaCiência SA, Lumiar, 1649‐038, Portugal
4Inhalation and Advanced Drug Delivery, R&D, Hovione FarmaCiência SA, Lumiar, 1649‐038, Portugal
Summary: The pulmonary route has been highly explored for drugs administration due to the advantageous features of lungs. DPI systems encompass a thorough mixing process, where the blending settings influence the resulting formulation characteristics. Carrier‐based formulations are widespread solutions for delivery of active ingredients to the lungs in the form of powder. Nevertheless, understanding the cohesive‐adhesive balances remains a challenge for pulmonary therapy. To further study this balance, eleven formulations were prepared through different order of addition of the components and different blending energies, to evaluate the impact of these factors on product performance. Statistically, there is no model that could present a viable explanation for the variations observed. However, results demonstrated that greater blending energy improved uniformity and homogeneity. It was recognized that the mixture of an appropriate amount of an excipient's pre‐blend with the API did not impact significantly, in order to establish an accurate correlation. The average content of API in each formulation from BUA analysis was demonstrated to be higher in trials with also a greater Fine particle fraction. DSC data was also obtained for these blends and allowed establishing a correlation between the amount of carrier fines and the drug delivery of the formulation. Therefore, and in agreement with the experimental results, a blending energy of 0.09 mJ/carrier (mixing time of X + 10 min) and the initial addition of the largest portion of pre‐blend of excipients to the API (Condition A) revealed to be optimal conditions to obtain a desirable uniformity and enhanced DPI performance.
In‐silico evaluation of different carrier particle geometries in interactive powder mixtures
Melvin Wostry and Regina Scherließ
Department of Pharmaceutics and Biopharmaceutics, Kiel University, Gutenbergstrasse 76, 24118 Kiel, Germany
Summary: In this study, an in‐silico model for the evaluation of different geometries as potential carrier particles in interactive powder mixtures was tested. Firstly, a carrier particle representing either a sphere or more complex geometries was loaded with an API and secondly, a collision of the loaded carrier particle with a wall was simulated to mimic the redispersion of API particles inside the inhaler during inhalation. In the loading simulations differences in the loading efficiency for different geometries were significant. The performance of the sphere with its high mass was in the average, in comparison the complex geometry of the rollingknot with its small mass could load four times more API related to its own mass. In the second step the effect of a particle‐wall collision was tested. In multiple setups the strength of adhesive forces between API and carrier as well as the type of the collision were altered to result in different extent of API adhesion/detachment on the surface. It was shown that at higher collision angles of up to 60° the detachment rate of API particles decreased whereas with higher collision velocities the detachment rate increased. Lower carrier surface energies resulted in overall higher detachment rates. The tested geometries showed the same trends under the same collision conditions, but nevertheless, the efficiency of the API detachment varied with the different carrier geometries. It was also shown that some geometries' performances were more dependent on the varied parameters while others showed a more constant detachment of the API particles.
Investigating the Dispersion Capability of High Dose DPI Formulations Manufactured Through Isothermal Dry Particle Coating (iDPC™)
Rhys Jones1, Amandip Gill1, David Wyatt1,2, Afzal Mohammed1,2 & Jasdip S Koner1
1Aston Particle Technologies, Aston Triangle, Birmingham, B4 7ET, United Kingdom
2Aston University, School of Life & Health Sciences, Birmingham, B4 7ET, United Kingdom
Summary: Isothermal Dry Particle Coating (iDPC) technology is a proprietary powder blending/coating technology highly suitable for the development and manufacture of DPI formulations The object of this study was to investigate the delivery performance of concentrated dispersions (containing 75%w/w active material) in a range of in different size Monodose RS01™ devices and at different capsule fill volumes. The active materials investigated were micronised fluticasone propionate (FP) and micronised salmeterol xinafoate (SX) which were dispersed by iDPC separately in alpha‐lactose monohydrate (Lactohale® 100 (LH100)) and in spray‐dried mannitol (Pearlitol® 200 SD (P200SD) model carriers. Aerodynamic particle size distribution data for all formulations demonstrated the iDPC uniformly dispersed FP and SX across both carriers, with highly efficient delivery observed. Fine particle doses demonstrated linear increases when FP and SX was formulated with lactose for all delivery systems. A similar pattern was observed with mannitol based formulations for both FP and SX with size 2 and 3 capsules, with relative doses being higher than lactose. However at size 0 a large reduction in emitted dose was observed at 60% fill volume, FPF performance was high across the board, highlighting the capability of iDPC to develop highly efficient high dose DPI formulation. Lactose based formulations highlighted a negative correlation to fill volume, with higher volumes lowering FPF. Mannitol based formulations displayed no particular pattern, with FPF remaining high even at 60% fill volume where FPD performance was disrupted. This work demonstrates that when concentrated iDPC formulations are coupled to the appropriate carrier and Monodose RS01 architecture excellent DPI performance can be achieved.
Assessment of the Automatic Filling of Orbital™ Device Loaded with a Model High Payload Blend Manufactured by Isothermal Dry Particle Coating
Cristina Rey‐Blanes & Reanne Beaird1, Norma Ulrich2, Katarzyna Epeslidis2, Niall Doherty1, Antonia Zapata del Baño1, Jonathan Tournaire3, Jonathan Mulpas3, Jasdip Koner4, Rhys Jones4, Andrea Silvestri5, Irene Rossi1
1Nanopharm Ltd, An Aptar Pharma Company, Grange Road, Cwmbran, NP44 3WY, United Kingdom
3Aptar Pharma SAS, Route des Falaises, Le Vaudreuil, 27100, France
4Aston Particle Technologies, Aston Triangle, Birmingham, B4 7ET, United Kingdom
5Sterling, Via della Carboneria 30, Corciano (PG), 06073, Italy
Summary: Orbital™ is a unit dose dry powder inhaler capable of delivering high doses to the lungs over several inhalations. The assessment of the realistic performance of Orbital™ filled manually or automatically using the Drum Lab was carried out. Isothermal Dry Particle Coating (iDPC) was used to blend (50:50% w/w) of lactose LH200 and a model cohesive drug (Fluticasone Propionate), based on previous results obtained. Two puck configurations were employed: 0.6 and 0.9 mm with a single orifice, together with two different fill weights, 125 and 250 mg. Puck content uniformity performed on the drum filled pucks confirmed homogeneity of the filling. Generally, the in vitro realistic aerodynamic performance (DD and FPM) was higher for the drum filled samples, especially for the first inhalation act. In addition, larger puck orifice translated into a higher DD and FPM which was reflected by the higher mass deposited in the different lung generations of the in silico model (alveolar > tracheobronchial > bronchiolar), only when the 0.9 mm puck was automatically filled. Finally, the positive influence of drum filling determined a higher cumulative mass emitted after five inhalations with the highest value collected for the 0.9 mm orifice puck at both fill weights (> 65%). These results demonstrated that drum is a feasible filling technology for the Orbital™ device in combination with an iDPC formulation. The iDPC blend proved to be easily handled, regardless of its high payload and poor flow characteristics, reporting even better aerodynamic results when automatic filling was performed.
