Dissertations / Theses on the topic 'Nasal dry powder inhaler'

The overall objectives of this research project were i) to develop and evaluate methods of characterizing nasal spray products using realistic nasal airway models as more clinically relevant in vitro tools and ii) to develop and evaluate a novel high-efficiency antibiotic nanoparticle dry powder formulation and delivery device. Two physically realistic nasal airway models were used to assess the effects of patient-use experimental conditions, nasal airway geometry and formulation / device properties on the delivery efficiency of nasal spray products. There was a large variability in drug delivery to the middle passages ranging from 17 – 57 % and 47 – 77 % with respect to patient use conditions for the two nasal airway geometries. The patient use variables of nasal spray position, head angle and nasal inhalation timing with respect to spray actuation were found to be significant in determining nasal valve penetration and middle passage deposition of Nasonex®. The developed test methods were able to reproducibly generate similar nasal deposition profiles for nasal spray products with similar plume and droplet characteristics. Differences in spray plume geometry (smaller plume diameter resulted in higher middle passage drug delivery) were observed to have more influence on regional nasal drug deposition than changes to droplet size for mometasone furoate formulations in the realistic airway models. Ciprofloxacin nanoparticles with a mean (SD) volume diameter of 120 (10) nm suitable for penetration through mucus and biofilm layers were prepared using sonocrystallization technique. These ciprofloxacin nanoparticles were then spray dried in a PVP K30 matrix to form nanocomposite particles with a mean (SD) volume diameter of 5.6 (0.1) µm. High efficiency targeted delivery of the nanocomposite nasal powder formulation was achieved using a modified low flow VCU DPI in combination with a novel breathing maneuver; delivering 73 % of the delivered dose to the middle passages. A modified version of the nasal airway model accommodating Transwell® inserts and a Calu-3 monolayer was developed to allow realistic deposition and evaluation of the nasal powder. The nanocomposite formulation was observed to demonstrate improved dissolution and transepithelial transport (flux = 725 ng/h/cm2) compared to unprocessed ciprofloxacin powder (flux = 321 ng/h/cm2).

Dry powder inhalers (DPI) are a common asthma treatment. Despite the number of commercial devices available, little is known about their internal operation: the process of fluidising a powder dose into an inhalation airflow. This PhD aims to investigate this process, and demonstrate that it can be modelled computationally. . Experimental work is described to record high speed video of the dose fluidisation from simplified DPls. Typical DPI powders such as lactose are tested, along with cohesionless glass spheres and aluminium flakes. Two distinct dose fluidisation mechanisms are identified, labelled 'fracture' and 'erosion'. Lactose exhibits a fracture mechanism -- large agglomerates are produced as the powder bed cracks along lines of weakness. Glass or aluminium particles exhibit an erosion mechanism: powder is entrained into the flow as individual particles from the bed surface. The recorded video is quantitatively analysed to determine fluidisation timescales and pressures. Shear cell test results show that predicting the mechanism of fluidisation is not possible using averaged bulk powder properties. This suggests any DPI model must include the fundamental particle interactions. The discrete element method (OEM) is introduced as a computational technique capable of predicting DPI behaviour from individual particle properties. The numerical accuracy of the method is assessed, showing that time integration is limited to a maximum of 2nd order accuracy due to discontinuities in particle contact forces. A sensitivity analysis shows inter-particle cohesion is the dominant factor affecting OEM predictions. OEM is used to create a simple model of the dose fluidisation that occurs within a DPI. The results are compared with real powder behaviour. OEM is shown to capture the realistic fluidisation of both lactose and glass powder doses. It is concluded that OEM is a promising technique to predict DPI behaviour, although further work is required to quantify inter--particle cohesive parameters

Dry powder inhalers (DPIs) are a common therapeutic modality for lung diseases such as asthma, but they are also used to treat systemic diseases such as diabetes. Advantages of DPIs include their portable design and low manufacturing costs. Another advantage of DPIs is their breath activation, which makes them popular among patients. In a passive DPI drug is only released when the patient inhales. When the patient inhales, air flows through the device. The flow of air entrains a dry powder formulation inside the device and carries it to the lung. Currently, no DPI exists which can deliver drug independent of the patient to the desired target site in the lung. This is because drug release depends on the patient’s inhalation manoeuvre. To maximize the effect of the treatment it is necessary to optimize DPIs to achieve drug delivery that (A) is independent of the inhalation manoeuvre and (B) is targeted to the correct site in the lung. Therefore, this thesis aims to apply numerical and experimental methods to optimize DPIs systematically. First, two clinically justifiable cost functions have been developed corresponding to the DPI design objectives (A) and (B). An Eulerian-Eulerian (EE) computational fluid dynamics (CFD) approach has then been used to optimize a DPI entrainment geometry. Three different optimized entrainment geometries have been found corresponding to three different therapeutic applications. Second, the CFD approach has been validated experimentally. This is the first experimental study to validate an EE CFD approach for DPI modelling. Third, a personalized medicine approach to DPI design has been proposed. The development of this approach makes it possible to achieve the design objectives for patients with highly different lung functions. Finally, an adaptive DPI with a variable bypass element has been developed. This DPI achieves design objectives (A) and (B) for patients with highly different lung functions with a single device. In contrast to the personalized medicine approach, there is no need to select the optimal amount of bypass, since the device adapts automatically.

Candidate active pharmaceutical ingredients (APIs) are routinely tested to determine such parameters as physical stability, chemical stability, and bioavailability. Preformulation analysis of APIs does not currently attemept to determine whether they will perform to an acceptable level once they have been formulated. In practice, the APIs are subjected to extensive in vitro testing of their performance in a formulation, combined with optimisation of the formulation. This formulation testing is both time-consuming and expensive. In the field of pulmonary drug delivery from dry powder inhalers (DPIs), the API has to be aerosolized effectively in order to penetrate the lunfs and reach its deposition target. In a conventional ternary DPI fromulation, the API is combined with carrier lactose and fine lactose particles. The inter-particle forces between these three components and the bulk properties of the formulation determine the structure of the formulation and the aerolization performance of the API. In this study, physicochemical properties of salbutamol base and several of its salts were investigated both quantitatively and qualitatively. The in vitro deposition characteristics of the formulated APIs were also determined. The relationship between these parameters and the deposition was analysed to establish if a rapid preformulation screening technique could be applied to the APIs with respect to predicting the deposition performance of the formulated API. A clear relationship between the deposition of the unformulated API and the formulated API was observed that could be exploited as a screening technique.

When it comes to dry powder inhalation (DPI), adhesive mixtures are the most widely used formulation type. Various techniques have been developed to generate inhaled drug particles and improve the delivery efficiency of DPI formulations. For dry powder inhaler formulations (DPIs), micronized drug powders are usually mixed with lactose carriers to improve powder handling during manufacturing and powder aerosol delivery during patient use. The performance of DPI systems is strongly dependent on several formulation factors, the construction of the delivery device and the inhalation technique. There is a growing interest in DPI in new medical areas such as vaccines and antibiotics which requires further development and challenges to ensure physical and aerosolization stability of DPI.  This project aims to discuss the development of inhalation therapy, the challenges during formulation processes, the mixing process and the use of excipients in pulmonary drug delivery in DPIs. Further, the project is covered by experiments based on the literature overview and performed at the Department of Pharmaceutical Biosciences at Uppsala University. Bulk density was measured on three series of adhesive mixtures with increasing amounts of fine particles. In two series, small amounts of Magnesium Stearate, 0,1% and 0,01% were added.

Dry powder inhalers (DPIs) are used for delivering drugs to the airways. In addition to the drug, the formulations often contain a coarse carrier, most commonly alpha lactose monohydrate. The presence of fine lactose particles in the formulation is known to improve the formulation performance. The active site, drug-fines agglomeration and increased cohesion theories have been suggested to explain improved DPI performance upon addition of fine excipient particles. This project aimed to investigate the validity of those theories. The viability of the active sites theory in explaining the improved DPI performance was investigated by studying the impact of loaded drug dose on the in vitro performance for formulation series prepared with coarse carriers with different surface characteristics. The formulations prepared with the rougher lactose carrier were seen to outperform the formulations prepared with the smoother carrier at all drug concentrations. These findings were concluded to be non-compatible with the active sites theory. The impact of addition of lactose fines with different size distributions on powder flow and fluidisation properties and in vitro performance was studied. Powder cohesion increased independent of size distribution of the fines, but did not necessarily correspond to improved performance. Therefore, the increased cohesion theory was concluded not to be the sole explanation for the improvement in DPI performance in the presence of lactose fines. Instead, the increase in performance could be preliminarily attributed to the formation of agglomerated systems. The formation and co-deposition of drug-fines agglomerates, and consequential improvement in the DPI performance was proved using morphologically directed Raman spectroscopy. The project also aimed to develop a universal model for predicting DPI performance based on the lactose properties for a wide range of carriers with different properties. No simple linear correlations between any the lactose properties and the final DPI performance were found. Therefore no single parameter can be used as a universal predictor for DPI performance. To establish more complex relationships, artificial neural networks were used for modelling the importance of different lactose properties in determining DPI performance. The proportion of fine lactose particles (<4.5 μm) was identified as the most important parameter. However, this parameter was capable of explaining only approximately half of the variation seen in the formulation performance. The current study showed that to obtain more accurate predictions for the purposes of quality-by-design approach, also other lactose properties need to be characterised.

Dry powder inhalers (DPIs) are well established as a means of delivering inhaled drugs to the lungs. Combination DPIs, containing two or more drugs in a single formulation, may improve patient compliance by simplifying medication regimens. The implications of co-formulation on the aerosolisation behaviour of the component drugs, however, are not fully understood. Using salmeterol xinafoate (SX) and fluticasone propionate (FP) as model drug powders, a systematic approach was undertaken to better understand the consequences of combination formulation on drug dispersion for inhaled delivery. A dry dispersion laser diffraction analysis was developed to characterise the inherent dispersibility of bulk powders as well as de-agglomeration of DPI blends. Eight inhaled drug/excipient powders displayed different dispersibility (represented by the DA50, the dispersing pressure to achieve 50 % de-agglomeration; 0.23-1.45 Bar) and cohesivity (represented by the critical primary pressure (CPP); 1.0-3.5 Bar). Upon co-formulation (in the absence of a carrier) SX deposition in the Next Generation Impactor (NGI) became less efficient as the FP content increased (e.g. fine particle fraction (FPF) 33-18% recovered dose (RD) for SX:FP ratios 1:0-1:8). However, FP dispersion was generally unaffected when blended with SX (FPF 26-29 % RD, SX:FP ratios 0:1-8:1). This was attributed to the greater adhesivity and cohesivity of SX and FP, respectively, and changes in bulk blend structure and dispersibility. Intra-batch heterogeneity in particle properties within a bulk powder were also studied using solid state, calorimetric and inverse gas chromatographic analysis. The NGI was used to isolate aerodynamic size fractions which displayed distinct physicochemical and aerosolisation properties. For example, FP sub-populations had better dispersibility (FPF 32 vs. 19 % RD, respectively) whereas an SX sub-population had higher bulk and surface disorder and poorer dispersibility (FPF 21 vs. 33 % RD, respectively) compared to the bulk powders. Upon co-formulation (in the absence of a carrier) the fractions responded differently in terms of their aerosolisation behaviour compared to the unfractionated powders, indicating heterogeneity in the response to co-formulation within a bulk powder. When formulated with a carrier, however, there were no modifications to the dispersion (i.e. FPF) of unfractionated SX or FP upon co-formulation at different SX:FP ratios, however, a fraction of SX co-formulated with FP in a DPI blend exhibited changes to SX dispersion that were comparable to those in the absence of a carrier. The effect of co-formulation on SX and FP dispersion, therefore, was found to depend on the drug ratio, properties of the powder/fraction employed and the presence of a carrier.

Thesis (Ph. D.)--University of North Carolina at Chapel Hill, 2007.
Title from electronic title page (viewed Dec. 18, 2007). "... in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Department of Molecular Pharmaceutics." Discipline: Molecular Pharmaceutics; Department/School: Pharmacy.

Dry powder inhalers (DPIs) are advantageous for delivering medication to the lungs for the treatment of respiratory diseases because of the stability of the powders, relative low cost, synchronization of inhalation and dose delivery, and many design options that can be used for optimization. However, currently marketed DPIs are very inefficient in delivering medications to the lungs. This study has developed multiple new high efficiency DPIs for use with spray dried excipient enhanced growth (EEG) powder formulations based on the following platforms: capsule-based for oral inhalation, high-dose for oral inhalation, inline with 3D rod array dispersion, and inline with capillary jet dispersion. The capsule-based DPIs for oral inhalation implemented a 3D rod array for aerosol dispersion with optimal designs producing mass median aerodynamic diameters (MMADs) in the range of 1.3-1.5 µm and emitted doses in the range of 79-81%. Keys to inhaler success were the orientation of the capsule and inclusion of the 3D rod array. For the high-dose oral inhaler, performance was similar to the optimized capsule-based devices, while aerosolizing a much larger mass of powder. Surprisingly, removal of the fluidized bed of spheres improved performance producing a simple high dose device containing only a single dose sphere. The inline device using the 3D rod array was effective in producing particles of approximately 1.5 µm, at flow rates consistent with high flow therapy using a 1 L ventilation bag as the delivery mechanism. Using a capillary jet as the dispersion mechanism, further advances were made to allow for both delivery using a low volume (LV) of air and delivery in low flow therapy. This easily adaptable platform was able to produce a high quality aerosol out of a nasal cannula with an ED greater than 60% and a size (~2 µm) that should produce minimal extrathoracic losses. In conclusion, this study demonstrates (i) the design and optimization of DPIs capable of delivering EEG aerosols to the lungs using oral inhalation, (ii) the ability to deliver EEG aerosols using N2L aerosol administration, and (iii) the design of a new flexible LV-DPI device that is easily adaptable to multiple patients and delivery platforms, which are greatly needed in clinical environments.

Dry Powder Inhalers (DPIs) have great potential in pulmonary drug delivery; the granular powder, used as active ingredient in DPIs, is ozone friendly and the operation of DPIs ensures coordination between dose release and patient inhalation. However, the powder fluidisation mechanisms are poorly understood which leads to low efficiency of DPIs with 10-35 % of the dose reaching the site of action. The main aim of this thesis is to study the hydrodynamics of powder fluidisation in DPIs, using experimental and computational approaches. An experimental test rig was developed to replicate the process of transient powder fluidisation in an impinging air jet configuration. The powder fluidisation chamber was scaled up resulting in a two dimensional particle flow prototype, which encloses 3.85 mm glass beads. Using optical image processing techniques, individual particles were detected and tracked throughout the experimental time and domain. By varying the air flow rate to the test section, two particle fluidisation regimes were studied. In the first fluidisation regime, the particle bed was fully fluidised in less than 0.25 s due to the strong air jet. Particle velocity vectors showed strong convective flow with no evidence of diffusive motion triggered by inter-particle collisions. In the second fluidisation regime, the particle flow experienced two stages. The first stage showed strong convective flow similar to the first fluidisation regime, while the second stage showed more complex particle flow with collisional and convective flow taking place on the same time and length scales. The continuum Two Fluid Model (TFM) was used to solve the governing equations of the coupled granular and gas phases for the same experimental conditions. Sub-models for particle-gas and particle-particle interactions were used to complete the model description. Inter-particle interactions were resolved using models based on the kinetic theory of granular flow for the rapid flow regime and models based on soil mechanics for the frictional regime. Numerical predictions of the first fluidisation regime showed that the model should incorporate particle-wall friction and minimise diffusion, simultaneously. Ignoring friction resulted in fluidisation timing mismatch, while increasing the diffusion resulted in homogenous particle fluidisation in contrast to the aggregative convective fluidisation noticed in the experiments. Numerical predictions of the second fluidisation regime agreed well with the experiments for the convection dominated first stage of flow up to 0.3 s. However, later stages of complex particle flow showed qualitative discrepancies between the experimental and the computational approaches suggesting that current continuum granular models need further development. The findings of the present thesis have contributed towards better understanding of the mechanics of particle fluidisation and dense multiphase flow in DPI in particular, and particle bed fluidisation using impinging air jet in general. The use of TFM for predicting high speed convective granular flows, such as those in DPIs, is promising. Further studies are needed to investigate the form of particle-particle interactions within continuum granular flow models.

Malgré les progrès réalisées en matière de traitement et de diagnostic, le cancer du poumon demeure le plus répandu et le plus mortel dans le monde. La chimiothérapie conventionnelle, associant un composé de platine (cisplatine ou carboplatine) à un autre agent antinéoplasique est utilisée à quasiment tous les stades. Comme celle-ci est administrée par voie intraveineuse (IV), elle entraîne des effets secondaires systémiques importants dont certains sont dose- limitant (DLT) comme la néphrotoxicité pour le cisplatine ou la myélotoxicité pour le doublet carboplatine-paclitaxel. Par conséquent, ces agents sont administrés selon des cycles bien espacés pendant lesquels les tissus se rétablissent, et ce incluant la tumeur ;conduisant à une repopulation tumorale. En effet, une corrélation significative a été établie entre la concentration de platine dans les tumeurs pulmonaires et l’efficacité du traitement. Le but de ce travail était d’évaluer le potentiel de combiner une poudre sèche pour inhalation (CIS-DPI-50) avec le traitement de chimiothérapie IV, afin d’exposer la tumeur à l’agent cytotoxique de manière continue.La première partie de ce travail a permis de développer le CIS-DPI-50. Afin d’éviter qu’une haute concentration en cisplatine ne soit complètement solubilisée une fois dans les poumons, et afin d’assurer une exposition suffisante, il était essentiel de développer des formulations à libération contrôlée et à rétention pulmonaire suffisante. Ceci consistait en l’optimisation d’une formulation à base de microparticules lipidiques solides (CIS-DPI-TS) préalablement développée par Levet et al. Cette formulation a été reproduite afin d’évaluer son efficacité chez des souris greffées avec le modèle M109-HiFR (0.5 mg/kg, trois fois par cycle pendant deux cycles) et a démontré une survie similaire au CIS-IV (1.5 mg/kg, une fois par cycle pendant deux cycles). Cela a été effectué en (i) utilisant des excipients de grade pharmaceutique, reconnus comme sûrs (GRAS) (49,5 % (w/w) d’HCO et 0,5 % (w/w) de TPGS) selon un processus facilement transposable, et (ii) en augmentant la libération initiale afin d’améliorer la réponse antitumorale. Le CIS-DPI-50 a montré une performance aérodynamique prometteuse à des débits d’air différents (100 et 40 L/min) avec une fraction de particules fines par rapport à la dose délivrée (FPF_d) de 86 ± 1 % et de 74 ± 1 %, respectivement. La reproductibilité du procédé a été démontrée sur 3 lots différents et la stabilité maintenue pendant les 6 mois de stockage avec une FPF_d variant de 81,0 ± 0,6 % au T0 à 81 ± 2 % après 6 mois. Ceci était lié à (i) la stabilisation de la forme β de HCO et de l’état cristallin du cisplatine, et (ii) à la faible teneur en solvant résiduel (< 0,2 % w/w). De plus, cette formulation était caractérisée par une libération initiale plus marquée qu’avec CIS-DPI-TS ainsi que par des propriétés de libération contrôlée in vitro puisque 48 ± 2% ont été dissous en 2 h, vs. 35 ± 11 % pour CIS-DPI-TS et 76 ± 5 % pour les microcristaux de cisplatine non enrobés. Cela a été confirmé in vivo et a prouvé que le changement vers HCO a diminué le Tmax dans le sang de 120 min pour CIS-DPI- TS à 1 min pour le CIS-DPI-50. De plus, la rétention pulmonaire a été maintenue pendant 4 heures avec une aire sous la courbe (AUC) dans les poumons de 4 611 ± 932 ng.min.mg1 vs. 6 072 ng.min.mg-1 pour CIS-DPI-TS. Par conséquent, cette formulation a été choisie pour la suite des investigations.La deuxième partie de ce travail visait tout d’abord à évaluer la biodistribution après l’administration de CIS-DPI-50 à 0.5 mg/kg chez des souris greffées avec le modèle LLC1- Luc. Suite à cette administration, une exposition plus soutenue et dix fois plus élevée a été retrouvée dans les tumeurs par rapport au tissu sain (AUC0-∞ de 10 683 ± 5 826 ng.min.mg-1, vs. 1 071 ± 825 ng.min.mg-1, respectivement). Le deuxième objectif était de sélectionner le schéma d’administration du CIS-DPI-50 le plus adapté à sa combinaison avec la chimiothérapie IV. Le CIS-DPI-50 a été administrée 5 fois par cycle pendant deux cycles à 0,3, 0,5 et 1 mg/kg, ou à 0,5 mg/kg 3 fois par cycle pendant deux cycles. Après le premier cycle de traitement, aucune différence en termes de concentrations en platine n’a été observée dans les tumeurs ou dans les organes sains entre les groupes traités de manière répétée et ceux administrés une seule fois. Cependant, un cycle plus tard, toutes les concentrations en platine ont augmenté dans les organes sains et diminué dans les tumeurs. Ceci était lié à une augmentation de la taille tumorale d’un facteur de 23 entre les deux cycles (533 ± 23 mg vs. 23 ± 3 mg), ainsi qu’à la dégradation de l’état général des animaux. De plus, aucun des schémas n’a démontré une toxicité pulmonaire ou une efficacité. Cette efficacité limitée était liée à la faible sensibilité du modèle LLC1-Luc au cisplatine. Par conséquent, les schémas caractérisés par la plus faible dose cumulée (0,5 mg/kg trois fois par cycle et 0,3 mg/kg cinq fois par cycle) ont été sélectionnés afin d’évaluer leur efficacité chez des souris greffées avec le modèle M109-HiFR-Luc2. Une réduction significative de la taille tumorale dans les groupes traités (p < 0,0001) par rapport au groupe non traité a été observée ;confirmant la réponse de ce modèle au cisplatine. Cependant, aucune différence en termes de croissance tumorale (tendances similaires), de proportion de répondeurs (33 % pour les deux groupes) ou de survie (31 jours pour les groupes traités vs. 23 jours pour le groupe non traité) n’a été rapportée entre ces deux groupes. Par conséquent, le schéma le moins fréquent a été choisi pour éviter une éventuelle accumulation de platine et une atteinte rénale aigue (AKI).La troisième partie de ce travail avait pour but d’étudier la tolérance pulmonaire et rénale du CIS-DPI-50, du CIS-IV et de leurs combinaisons. Les résultats de quantification des cytokines pro-inflammatoires (TNF-α, IL-6, IL-1β) dans le fluide de lavage bronchoalvéolaire (BALF) ont montré une meilleure tolérance pour le CIS-DPI-50 par rapport au CIS-IV. Les neutrophiles granulocytes (NT-GRA) ont augmenté proportionnellement à la dose pour tous les groupes traités avec le CIS-DPI-50. Ces augmentations étaient réversibles une semaine plus tard uniquement pour les monothérapies et le groupe combiné, dont les administrations ont été espacées de 24h. Compte tenu des résultats d’inflammation et de cytotoxicité, l’ajout de CIS- DPI-50 au CIS-IV à sa dose maximale tolérée (MTD) semblait avoir un plus grand impact que si CIS-DPI-50 était ajouté à une dose IV réduite de 25%. Les résultats de quantification des biomarqueurs AKI plasmatiques (NGAL, cystatine C et créatinine) ont augmentés lorsque les deux monothérapies ont été administrées à leur DMT le même jour ou 24 heures plus tard. Par conséquent, la MTD du CIS-IV devait être réduite de 25% et les administrations séparées de 24h pour préserver la tolérance. L’efficacité de ce schéma a été évaluée sur des souris greffées avec le modèle M109-HiFR-Luc2 en combinant le CIS-DPI-50 au doublet IV cisplatine- paclitaxel. Malgré le fait que ces résultats n’étaient pas significativement différents, des tendances intéressantes en termes de réduction de la croissance tumorale, de survie (31 jours pour le groupe combiné vs. 26 pour le doublet IV, vs. 21 jours pour le groupe non traité) et de proportion de répondeurs (67 % pour le groupe combiné, vs. 50 % pour le doublet IV) ont été observés pour le groupe combiné.Comme les différentes adaptations ont probablement pu entraver le potentiel des combinaisons, il était intéressant d’étudier l’association de CIS-DPI-50 avec un doublet moins néphrotoxique (carboplatine-paclitaxel), et qui nécessiterait éventuellement moins d’ajustements. Compte tenu de la DLT du carboplatine et du paclitaxel, l’évaluation de la myélotoxicité était incluse dans cette étude. Les résultats ont montré que l’ajout de CIS-DPI-50 au doublet carboplatine- paclitaxel IV le même jour à leur MTD ont induit une augmentation du nombre de globules blancs et de cellules totales dans le BALF, une proportion plus élevée de NT-GRA dans le BALF et une anémie régénérative plus précoce qu’avec le doublet IV. Ces effets étaient réversibles. La stratégie de réduction de la dose IV de 25 % et la séparation des administrations par 24h ont permis d’éviter le développement d’une anémie régénérative et/ou l’augmentation de globules blancs ou du nombre de cellules totales dans le BALF par rapport aux doublets IV. De plus, toutes les combinaisons ont induit une cytotoxicité non réversible tout en étant mieux tolérées que celles avec le CIS-IV. Leurs efficacités devraient donc être testées sur des modèles de cancer pulmonaire murin seuls ou en combinaison avec l’immunothérapie (inhibiteurs de checkpoint).Ce travail a démontré la faisabilité de combiner une modalité de traitement locorégionale avec les traitements de chimiothérapie conventionnelle par voie IV à base de cisplatine et de carboplatine contre les tumeurs pulmonaires. Ceci a été effectué en optimisant les combinaisons afin d’éviter des toxicités pulmonaire, rénale et hématologique (pour la chimiothérapie à base de carboplatine) tout en démontrant une tendance vers une efficacité (pour le doublet cisplatine- paclitaxel) dans un modèle préclinique agressif. Par conséquent, ces résultats ouvrent la voie à plusieurs autres possibilités de combinaisons (traitements localisés et inhibiteurs de checkpoint) qui doivent être investiguées afin de sélectionner les indications pour lesquelles ce traitement serait le plus efficace.
Doctorat en Sciences biomédicales et pharmaceutiques (Pharmacie)
info:eu-repo/semantics/nonPublished

Chronic respiratory diseases such as asthma, COPD, pulmonary fibrosis are more prevalent throughout the world. For some of these diseases there is no cure, the current treatment options manages the symptoms and acute exacerbation. The new approach to find a curative therapy for respiratory diseases is by targeting the cellular / molecular pathways that either cause the disease or has the potential cure the disease. It becomes important to target the respiratory system in treating these diseases to increase the delivered dose and reduce the unwarranted adverse effects. Dry powder inhaler (DPI) is a targeted drug delivery dosage form commonly used to target the airways to treat respiratory diseases. There are two components to dry powder inhaler product – powdered drug formulation and inhaler device; a unified performance of the two is essential for a successful product. In this study, dry powder aerosol of novel drug compounds that targets the underlying cellular and molecular mechanism are developed for the first time. Advanced organic closed mode spray drying technique was used to the produce microparticulate/ nanoparticulate formulations. The formulation of the novel compounds involved utilizing sugar based excipients. Each formulation that was produced was comprehensively characterized in the solid state. The safety of these formulations were tested in in vitro human pulmonary cell lines. The in vitro aerosol dispersion of the spray dried drugs were tested using three FDA approved human inhaler devices. The influence of the inhaler device resistance and spray drying process conditions on the aerosol dispersion was evaluated. Preliminary testing of the formulations in in vivo animal models shows promising results in treating chronic respiratory diseases with these superior aerosol formulations.

The aim of the work was to design, manufacture, and characterize targeted multi-component dry powder aerosols of (non-destructive) mucolytic agent (mannitol), antimicrobial drug (tobramycin or azithromycin), and lung surfactant mimic phospholipids (DPPC:DPPG=4:1 in molar ratio). The targeted dry powder for inhalation formulation for deep lung delivery with a built-in rationale of specifically interfering several disease factors of chronic infection diseases in deep lungs such as cystic fibrosis, pneumonia, chronic bronchitis, and etc. The dry powder aerosols consisting of selected chemical agents in one single formulation was generated by using spray drying from organic solution. The physicochemical properties of multi-component dry powder inhaler (DPI) formulation were characterized by a number of techniques. In addition, the in vitro aerosol dispersion performance, storage stability test, and in vitro drug release of selected spray-dried (SD) multi-component systems were conducted. The physicochemical study revealed that multi-component aerosol particles possessed essential particle properties suitable for deep lung delivery. In general, the multi-component particles (typically 0.5 to 2 µm) indicated that the designed SD aerosol particles could potentially penetrate deep lung regions (such as respiratory bronchiolar and alveolar regions) by sedimentation and diffusion, respectively. The essential particle properties including narrow size distribution, spherical particle and smooth surface morphologies, and low water content (or water vapor sorption) could potentially minimize interparticulate interactions. The study of in vitro aerosol dispersion performance showed that majority of SD multi-component aerosols exhibited low values (less than 5µm) of MMAD, high values (approximately above 30% up to 60.4%) of FPF, and high values (approximately above 90%) of ED, respectively. The storage stability study showed that azithromycin–incorporated multi-component aerosol particles stored at 11 and 40% RH with no partial crystallization were still suitable for deep lung delivery. Compared to SD pure azithromycin particles, the azithromycin-incorporated multi-component particles exhibited an enhanced initial release. The targeted microparticulate and nanoparticulate multi-component dry powder aerosol formulations with essential particle properties for deep lung pulmonary delivery were successfully produced by using spray drying from organic solution. The promising experimental data suggest the multi-component formulations could be further investigated in in vivo studies for the purpose of commercialization.

The present work aimed at assessing the dose emission and aerodynamic particle size characteristics of formoterol fumarate from Atimos Modullite, a metered dose inhaler (MDI) and Foradil Aeroliser, Easyhaler, and Oxis Turbuhaler dry powder inhalers (DPI) at different inhalation flow rates and volumes using in vitro methodology. Recognised methods have been adopted and validated to generate the results. The in vitro characteristics of formoterol were measured according to standard pharmacopeial methodology with adaptation to simulate routine patient use. The dose emission from the Atimos Modulite was determined using inhalation volumes of 4 and 2 L and inhalation flows of 10, 28.3, 60, and 90 L/min. The %nominal dose emitted was consistent between the various flow rates and inhalation volumes of 4 and 2L. The particle size distribution was measured using an Anderson Cascade Impactor (ACI) combined with a mixing inlet valve to measure particle size distribution at inhalation flow rates below 30 L/min. The particle size distribution of formoterol from Atimos Modulite was measured using inhalation flows of 15, 28.3, 50, and 60 L/min with and without different spacers, Aerochamber and Volumatic. The mean fine particle dose (%nominal dose) through an Atimos without spacer were 53.52% (2.51), 54.1% (0.79), 53.37% (0.81), 50.43% (1.92) compared to Aerochamber 63.62% (0.44), 63.86% (0.72), 64.72% (0.47), 59.96% (1.97) and Volumatic 62.40% (0.28),63.41% (0.52), 64.71% (0.61), 58.43% (0.73), respectively. A small decrease in the fine particle dose was observed as the inhalation flow increased, but this was not significant. The respective mean mass aerodynamic diameter (MMAD) increased as the flow rate was increased from 15 of 60 L/min. Results also suggests that the use of spacers provides better lung deposition for patients with problems using MDI. The dose emission from the Foradil Aeroliser was determined using inhalation volumes of 4 and 2 L, at inhalation flows of 10, 15, 20, 28.3, 60, and 90 L/min plus two inhalations per single dose. The %nominal dose emitted using 2 L inhalation volume was approximately half when compared to results obtained using inhalation volume of 4 L. A significantly (p<0.001) higher amount of drug was also emitted from Easyhaler® at inhalation volume of 4 L through flow rates of 10, 20, 28.3, 40, and 60 L/min compared 2 L. Similar results were observed through Oxis Turbuhaler at inhalation flow rates of 10, 20, 28.3, 40, and 60 L/min. Comparative studies were also carried out to evaluate the particle size distribution of formoterol through the DPIs. The nominal fine particle dose through Aeroliser using inhalation flows of 10, 20, 28.3, 60 and 90 L/min were 9.23%, 14.70 %, 21.37%, 28.93%, and 39.70% for the 4 L and 4.17%, 5.55%, 7.28%, 8.41%, and 11.08% for the 2 L, respectively. The respective MMAD significantly (p<0.001) decreased with increasing flow rates. Aeroliser performance showed significant (p<0.001) increase in the % nominal fine particle dose for two inhalations compared to one inhalation at both 4 and 2 L. The Easyhaler was measured using inhalation flows of 10, 20, 28.3, 40, 60 L/min. The nominal fine particle dose were 19.03%, 27.09%, 36.89%, 49.71% and 49.25% for the 4 L and 9.14%, 15.44%, 21.02%, 29.41%, 29.14% for the 2 L, respectively. The respective MMAD significantly (p<0.001) decreased with increasing flow rates. Easyhaler performance at both 4 and 2 L showed no significant differences between one and two inhalations at low flow rates (10, 20, 28.3), but this was significant (p<0.05) at higher flow rates (40 and 60 L/min). The Oxis Turbuhaler was also measured using inhalation flows of 10, 20, 28.3, 40, 60 L/min. The nominal fine particle dose were 12.87%, 24.51%, 28.25%, 34.61%, 40.53% for the 4 L and 8.55%, 15.31%, 21.36%, 19.53%, 22.31% for the 2 L, respectively. Turbuhaler performance showed significant (p<0.05) differences between one and two inhalations at varying flow rates 2 L inhalation volumes, but not at 4 L. The use of Foradil Aeroliser delivers small particles as the Oxis Turbuhaler using two inhalations hence delivering formoterol deep into the lungs. Also, this thesis shows that high flow resistance of Turbuhaler will indeed influence the ability of patients with severe asthma or children to use the system. Beside, Easyhaler produced the highest drug delivery to the lungs, thus, making it a more desirable system to use, especially for children and asthma sufferers.

A asma é definida como uma doença inflamatória crônica de caráter multifatorial, caracterizada pela obstrução reversível das vias aéreas, denso infiltrado inflamatório e hiper-reatividade brônquica a estímulos externos. Clinicamente, a doença é marcada por sintomas episódicos de dispneia, sibilo, tosse seca e sensação de aperto no peito. A terapia convencional da asma compreende o uso de anti-inflamatórios e broncodilatadores. A budesonida é um glicocorticoide esteroide e é dos fármacos mais utilizados na terapêutica da asma. No entanto, a budesonida apresenta baixa biodisponibilidade oral e o uso prolongado pode levar a efeitos adversos graves como afinamento da pele e supressão adrenocortical. No desenvolvimento de novas formulações, a avaliação da toxicidade é de extrema importância. Por conseguinte, o uso de cultura celular é de grande valia no desenvolvimento de protocolos para avaliação da toxicidade de novas formulações. Adicionalmente, a nanotecnologia é uma ferramenta importante para resolver problemas de biodisponibilidade e para contornar efeitos adversos da terapêutica convencional. Desta forma, o objetivo desta tese foi desenvolver um novo sistema nanoestruturado na forma de pó seco para inalação (Dry powders inhalers – DPI), obtido por aspersão contendo budesonida encapsulada, visando o tratamento da asma aguda e crônica. Essa proposta foi baseada na obtenção de um sistema pulverulento nanoestruturado com tamanho reduzido e controlado, visando a entrega pulmonar da budesonida. Na etapa de pré-formulação foi realizado um estudo fatorial avaliando diferentes métodos de preparação das nanocápsulas e os adjuvantes de secagem utilizados. As análises de tamanho de partícula, da formulação selecionada (nanocápsulas contendo budesonida e secas por aspersão com leucina) mostraram um tamanho reduzido e adequado para a administração pulmonar (2,7 μm). A morfologia demonstrou que estas partículas possuem um tamanho reduzido, forma esférica e superfície irregular, características importantes para a administração pulmonar. Quando analisada a distribuição pulmonar in vitro, em Impactador de Andersen, a formulação apresentou uma fração de partículas finas (Fine Particle Fraction – FPF) de 28%. Analisando os resultados dos experimentos em modelos de asma aguda e crônica induzidos por ovalbumina, os resultados da mecânica respiratória e função pulmonar mostraram uma diminuição na resistência e na elastância pulmonar, quando a budesonida nanoencapsulada foi utilizada, quando comparada com uma formulação comercial de budesonida, nas duas doses utilizadas (0,5 e 1,0 mg/Kg). Esse tratamento com nanocápsulas também mostrou eficiência na redução da inflamação, pela redução do número de leucócitos totais no fluido de lavagem bronco alveolar (Broncho Alveolar Lavage Fluid – BALF) e, principalmente, redução significativa no número dos eosinófilos no infiltrado pulmonar. Corroborando esses resultados, a quantificação da eotaxina – 1 e das citocinas pró-inflamatórias foram reduzidas, quando comparadas ao tratamento comercial. A análise histopatológica mostrou que quando o tratamento com as nanocápsulas foi utilizado, a produção de muco foi reduzida, bem como a produção de fibrose sub-epitelial, sugerindo um possível efeito sobre o remodelamento tecidual. Os resultados de toxicidade utilizando linhagem celular epitelial pulmonar (H441) mostrou uma redução na toxicidade da budesonida, quando encapsulada nas nanopartículas, tanto na forma de suspensão como na forma pulverulenta. Essa redução da toxicidade foi de 75% e de 50%, na dose de 100 μg/mL, para a suspensão e para o DPI, respectivamente. O conjunto dos resultados obtidos mostrou a potencial aplicabilidade da budesonida nanoencapsulada para o tratamento da asma, utilizando esse novo sistema DPI.
Asthma is characterized as a chronic inflammatory disease developed by multifactorial aspects such as genetic predisposition and exposure to environmental factors such as pollution, smoke and microorganisms. The conventional asthma therapy comprises the use of bronchodilators and anti-inflammatory. Budesonide is a glucocorticoid and is the most frequently used therapy in the treatment of asthma. However, this drug has low oral bioavailability and long term use may lead to adverse effects such as skin thinning and adrenal suppression. The evaluation of the toxicity of new formulation has critical role in the pharmaceutical development. The use of cell culture experiments can help this aspect. Additionally, nanotechnology is an important tool to solve problems regarding bioavailability and to circumvent adverse effects of conventional therapy. The aim of this work was to develop a nanostructured system as dry powder inhaler (DPI) containing budesonide loaded, obtained by spray-drying, targeting the treatment of acute and chronic asthma. This proposal was based on obtaining a nanostructured powder system with reduced and controlled size, aiming an alternative to treatment of asthma. A factorial study comparing different methods to produce the nanocapsules as well as the type of drying adjuvants was performed. The particle size of the selected formulation was 2.7 μm, an adequate reduced size suitable for pulmonary administration. The morphology of these particles showed a small size, spherical shape and irregular surface. All these characteristics are important for pulmonary administration. When analyzed the in vitro pulmonary distribution of the DPI, using an Andersen Cascade Impactor, showed a fine particle fraction (FPF) of 28%. Analyzing the results of the biological experiments, the mechanical respiratory and pulmonary function showed a decrease in lung elastance and resistance when budesonide was used nanoencapsulated compared with a commercial formulation of budesonide in two doses (0.5 and 1.0 mg / kg). Both treatments also showed nanocapsules efficiency in reduction of inflammation by reducing the total of leukocytes in the bronchial alveolar lavage fluid (BALF) and especially significant reduction in eosinophil infiltration in the lung tissue. Corroborating with these results, the quantification of eotaxin - 1 and proinflammatory cytokines was reduced when compared to commercial budesonide treatment. Histopathological analysis showed that when treatment with the nanocapsules was used, mucus production was reduced and reversed the phenomena of airway remodeling. The cytotoxicity assay by Alamar blue using the bronchial epithelium cell line (H441) showed a reduction on the toxicity of budesonide when the nanocapsules were used even in suspension or in the DPI. The cytotoxicity reduction were 75 and 50%, at 100 μg/mL, for the suspension and the DPI, respectively. All these results show that budesonide-loaded nanocapsules in dry powder inhaler is a promising approach for the treatment of asthma.

This research was aimed at the development and validation of new in vitro methods capable of predicting in vivo drug deposition from dry powder inhalers, DPIs, in lung-normal human adults. Three physical models of the mouth, throat and upper airways, MT-TB, were designed and validated using the anatomical literature. Small, medium and large versions were constructed to cover approximately 95% of the variation seen in normal adult humans of both genders. The models were housed in an artificial thorax and used for in vitro testing of drug deposition from Budelin Novolizer DPIs using a breath simulator to mimic inhalation profiles reported in clinical trials of deposition from the same inhaler. Testing in the model triplet produced results for in vitro total lung deposition (TLD) consistent with the complete range of drug deposition results reported in vivo. The effect of variables such as in vitro flow rate were also predictive of in vivo deposition. To further assess the method’s robustness, in vitro drug deposition from 5 marketed DPIs was assessed in the “medium” MT-TB model. With the exception of Relenza Diskhaler, mean values for %TLD+SD differed by only < 2% from their literature in vivo. The relationship between inhaler orientation and in vitro regional airway deposition was determined. Aerosol drug deposition was found to depend on the angle at which an inhaler is inserted into the mouth although the results for MT deposition were dependent on both the product and the formulation being delivered. In the clinic, inhalation profiles were collected from 20 healthy inhaler naïve volunteers (10M, 10F) before and after they received formal inhalation training in the use of a DPI. Statistically significant improvements in Peak Inhalation Flow Rate (PIFR) and Inhalation Volume (V) were observed following formalized training. The shapes of the average inhalation profiles recorded in the clinic were found to be comparable to the simulated profiles used in the in vitro deposition studies described above. In conclusion, novel in vitro test methods are described that accurately predict both the average and range of aerosol airway drug deposition seen from DPIs in the clinic.

An issue of inhalation therapy is a complex topic, actively discussed in last decades, and its progress in various scientific fields is more than required. First part of this thesis brings a theoretical introduction into principles of aerosol therapy and into the requirements resulting from them. Commonly available technologies of inhalers and nebulisers for medical usage, parameters that determinate their effectivity are briefly described. Usage mistakes influencing the effectivity of inhalation are discussed, as well. Second part deals with experimental measurements of aerosol that selected inhalers generate. It also describes difficulties connected with the methods of these measurements, with sampling and following analyses. Gained results are compared with an available literature.

Les tumeurs pulmonaires, qu’elles soient primaires (principalement représentées par le cancer du poumon non-à-petites cellules) ou secondaires (métastases), causent la mort de plusieurs centaines de milliers de personnes par an à travers le monde. Malgré les modalités de traitements existantes, un plateau thérapeutique a été atteint avec un taux de survie à 5 ans de maximum 15%. Actuellement, il est connu que le cancer du poumon non-à-petites cellules ainsi que les métastases sont intrinsèquement résistants à l’apoptose.

Pour apporter des réponses aux principales problématiques rencontrées avec l’administration systémique de la chimiothérapie conventionnelle qui est principalement constituée d’agents pro-apoptotiques, nous avons développé des formulations à base d’agents antinéoplasiques aux propriétés anticancéreuses non pro-apoptotiques qui sont destinées à être administrées de manière localisée par la voie inhalée. Il faut savoir que l’inhalation est la voie d’administration préférentielle des principales affections respiratoires telles que l’asthme, la bronchopneumonie chronique obstructive et la mucoviscidose.

La première partie de ce travail a consisté à produire et à évaluer des formulations à base de témozolomide destinées à être administrées chez la souris porteuse de pseudo-métastases pulmonaires (issues d’un mélanome expérimental, le modèle B16F10), soit via la voie intraveineuse (iv) conventionnelle soit via la voie inhalée à l’aide d’un dispositif endotrachéal approprié. La suspension pour inhalation a été produite à l’aide de technique de réduction de taille et a été stabilisée à l’aide de phospholipides compatibles avec la voie pulmonaire. L’activité anticancéreuse in vitro a été vérifiée pour le témozolomide formulé sous forme de suspension pour inhalation et de solution intraveineuse par rapport à du témozolomide non formulé sur des lignées de cellules cancéreuses de cancer humain NSCLC A549, de glioblastome humain T98G et de mélanome murin B16F10. Cette dernière lignée a été utilisée pour générer les pseudo-métastases pulmonaires chez la souris en injectant les cellules de mélanomes dans la voie systémique via la veine caudale. La reproductibilité de la dose et de l’aérosol générés à partir de la suspension pour inhalation à l’aide du dispositif d’administration endotrachéal et la déposition des gouttelettes dans les poumons de la souris ont pu être respectivement évaluées avec précision par une méthode de quantification du témozolomide qui a été validée par nos soins, par des techniques de diffraction laser et par des techniques de microscopie à fluorescence et d’analyse d’images histologiques. Enfin, l’activité antitumorale in vivo et la tolérance des traitements conventionnels ou localisés ont été vérifiées chez la souris porteuse de ces pseudo-métastases pulmonaires B16F10. Les résultats ont montré que le dispositif endotrachéal utilisé permettait de produire des doses et des aérosols reproductibles et de déposer les gouttelettes d’aérosol profondément dans les poumons des souris. De plus, lors de l’étude in vivo, les traitements administrés étaient bien tolérés et la dose de témozolomide administré sous forme de suspension pour inhalation à l’aide du dispositif endotrachéal avait permis d’obtenir une efficacité antitumorale similaire à une dose similaire de témozolomide administrée par la voie iv conventionnelle. De plus, 11% (3/27) de souris « long-survivantes » avaient été observées avec le groupe traité par inhalation trois fois par semaine pendant trois semaines consécutives et les poumons de ces long-survivants avaient présenté une éradication quasi complète des tumeurs pulmonaires. Ce phénomène n’avait pas été observé dans les groupes de souris traitées de manière conventionnelle.

Ensuite, la seconde partie de notre travail a consisté en l’élaboration du témozolomide sous forme de poudres sèches pour inhalation destinées à être délivrées à l’aide d’un dispositif à poudre sèche à usage humain. Pour ce faire, nous avons développé les poudres sèches pour inhalation à l’aide de techniques de réduction de taille pour microniser la poudre de départ et d’atomisation pour évaporer le solvant et élaborer un enrobage autour des particules micronisées. La nature de l’enrobage était soit hydrophile soit lipophile. Ensuite les caractéristiques physicochimiques telles que les propriétés thermiques, les propriétés cristallines, la distribution de taille particulaire et la morphologie des formulations de poudre sèche pour inhalation ont été évalués à l’aide de techniques appropriées telles que la calorimétrie différentielle à balayage, la diffraction des rayons X sur poudre, la diffraction de la lumière laser et la microscopie électronique à balayage. Les profils de déposition pulmonaire et de dissolution ont été respectivement déterminés in vitro à l’aide de l’essai de la pharmacopée européenne utilisant l’impacteur à cascade multi-étages et d’un test de dissolution adapté aux formes pulmonaires. Les quatre formulations élaborées présentaient des caractéristiques physicochimiques intéressantes pour la stabilité à long-terme de la substance active et des formulations. De plus, deux formulations de poudre sèche pour inhalation (les formulations F1 et F2) présentaient des propriétés aérodynamiques tout à fait attrayantes avec une fraction minimale de poudre déposée au niveau du tractus respiratoire supérieure et une fraction maximale de poudre déposée au niveau du tractus respiratoire inférieur où se localisent les tumeurs pulmonaires. De plus, l’ensemble des formulations ont montré que la fraction sélectionnée des particules fines des poudres sèches pour inhalation libérait 75% du témozolomide dans le liquide simulant le fluide pulmonaire endéans les dix premières minutes du test de dissolution in vitro adapté aux formes pulmonaires.

Enfin, nous avons comparé l’efficacité et la tolérance in vivo d’une de nos formulations de poudre sèche de témozolomide pour inhalation administrée soit sous forme de suspension, soit sous forme de poudre sèche, à l’aide du dispositif endotrachéal approprié chez la souris porteuse de pseudo-métastases pulmonaires. L’uniformité de la dose délivrée par les différents dispositifs a été évaluée à l’aide d’une technique quantitative validée. Les résultats de cette étude ont montré qu’en administrant une formulation de poudre sèche sous forme d’un mélange de poudres plutôt que sous forme d’une suspension liquide, les doses en témozolomide à administrer pour obtenir une efficacité comparable était au moins deux fois moins élevées. Cependant, le dispositif endotrachéal pour les formulations de poudre présentait plus de variabilité au niveau de la dose délivrée que le dispositif endotrachéal pour les formulations liquides ce qui induit une variabilité dans les doses délivrées. Pour clôturer ce travail, nous avons appliqué certaines techniques que nous avons développées pour le témozolomide à une nouvelle molécule de synthèse, le trivanillate polyphénolique 13c, qui montre des propriétés anticancéreuses intéressantes dans le cadre des tumeurs pulmonaires. En effet, une méthode quantitative a été développée et a été validée. Une étude de pré-formulation et des essais de formulation pour produire une suspension, des complexes d’inclusion et des microparticules lipidiques ont été entrepris avec de relativement bons résultats pour les complexes d’inclusion élaborés avec des gamma cyclodextrines qui permettaient d’augmenter la solubilité dans l’eau de la molécule de 13c d’un facteur d’au moins 1,5×106. De plus, les particules de 13c montraient la particularité de se solubiliser dans un mélange dichlorométhane/éthanol (1 :1 v/v) ce qui nous a permis d’élaborer des microparticules lipidiques pour lesquelles les propriétés de mouillage devront être améliorées dans l’avenir./

Primary lung tumors, mainly represented by non-small-cell lung cancers (cancers NSCLC), or secondary lung tumors (metastasis) cause the death of hundred thousand human beings worldwide. Despite the therapeutic modalities used, the five-year survival rate reaches only 15%. Nowadays, it is known that cancers NSCLC and metastasis are intrinsically resistant to apoptosis.

To overcome the main problems occurring with the systemic delivery of conventional chemotherapy which are mainly constituted of non-specific and non selective pro-apoptotic agents, we have developed some formulations based on non pro-apoptotic antineoplasic drugs which are designed to be delivered by a localized administration, the inhalation. Indeed, inhalation is the preferential route to treat the main pulmonary affections such as asthma, chronic obstructive pulmonary disease or cystic fibrosis.

The first part of this work consisted to produce and evaluate temozolomide-based formulations designed to be delivered to mice bearing pulmonary pseudo-metastases (using a experimental melanoma, the B16F10 model), either by the conventional intravenous (iv) route or by inhalation using an endotracheal device appropriate to mice. The suspension for inhalation was produced by means of a high pressure homogenizing technique using phospholipids compatible with the lungs to stabilize the suspension. The in vitro anticancer activity was evaluated for both temozolomide-based formulations in comparison with non-formulated temzolomide on three cancer cell lines, a human NSCLC cancer cells (A549), a human glioblastoma cancer cells (T98G) as positive control and a murine melanoma cancer cells (B16F10). The latter was used to generate lung tumors in mice by injecting the melanoma cells by iv. Reproducibility of delivered dose and generated aerosol by the endotracheal device from the suspension for inhalation and the deposition of droplets in the mouse lungs were precisely evaluated by means of a validated HPLC determination method, a laser diffraction technique and fluorescent microscopy and histological image analysis, respectively. Then, the tolerance and the antitumor efficacy of iv or inhaled temozolomide-based treatments were evaluated on mice bearing pulmonary pseudo-metastases B16F10. The results showed that endotracheal device produced reproducible doses and aerosols and that the aerosol droplets were deposited deeply in the mouse lungs. Moreover, the temozolomide-based treatments were well tolerated in terms of weight evolution and the inhaled based-temozolomide treatments were able to get the same antitumor efficacy in terms of median survival rate as the conventional iv based-temozolomide treatments delivered at a same frequency. Moreover with the group treated by inhalation three times a week during three consecutive weeks, 11% (3/27) mice survived with an almost complete eradication of lung tumors which was not observed with the groups treated by conventional route.

Then, the second part of our work consisted to produce temozolomide-based dry powders for inhalation able to be delivered with a dry powder inhaler for human use. We developed the dry powders for inhalation using a high-pressure homogenizing technique to micronize temozolomide particles and then spray-drying technique to coat temozolomide microparticles. The coating was either hydrophilic or lipophilic. Then, the physicochemical characteristics such as thermal or crystalline properties, the particle size distribution and the particle morphology were evaluated for the four dry powders for inhalation by means of differential scanning calorimetry, x-ray powder diffraction, laser light scattering and scanning electron microscopy, respectively. The in vitro pulmonary deposition and dissolution were respectively determined by European pharmacopeia assay for the aerodynamic assessment of fine particles using a multi-stage liquid impinger and by dissolution test optimized for inhaler products. The four formulations produced presented physicochemical properties promoting long-term stability of temozolomide and formulations.Moreover, two of them (dry powder without coating or with a thin lipid coating) showed attractive aerodynamic properties with a minimal fraction of powder deposited in the oropharyngeal and tracheal zones and maximal fraction deposited in the lungs (almost 50% of the nominal dose) where the lung tumors are localized. Moreover, fine particle fraction of all formulations showed a fast release and dissolution of temozolomide with more than 75% of temozolmide dissolved within 10 minutes in the simulated lung fluid during the in vitro dissolution test optimized for dry powders for inhalation.

Then, we compared the in vivo antitumor efficacy and tolerance of one of dry powders for inhalation on mice bearing pulmonary pseudo-metastases B16F10. The dry powder for inhalation was administered either by dispersing it as a extemporaneous suspension able to be delivered by the endotracheal device for liquid forms or by mixing it with a spray-dried diluent able to be delivered by the endotracheal device for dry powders. The uniformity of delivered dose by the different endotracheal device was evaluated by a validated quantitative method. The results showed that the delivery of the powder mixture presented the same antitumor efficacy as the extemporaneous suspension but for a half dose of temozolomide. However, the endotracheal device for dry powders presented a higher variability of delivered dose than the endotracheal device for liquid forms.

Finally, we apply the pulmonary application on a polyphenol developed in the Faculty of Pharmacy, the molecule 13c, that showed very interesting in vitro anticancer properties against lung tumors. So, a quantitative method was developed and was validated. A preformulation studie was performed and formulation developements are on-going.

Doctorat en Sciences biomédicales et pharmaceutiques
info:eu-repo/semantics/nonPublished

Lung cancer is the deadliest cancer in the world, with a global 5-year survival rate of about 15%. Despite a notable impact of the latest improvements in prevention, screening, detection and staging, the efficacy of conventional treatments is not sufficient and has reached a therapeutic plateau. These conventional treatments involve a combination of surgery, radiotherapy (RT) and chemotherapy (CT). CT is used in almost all stages: in operable and inoperable stages to limit tumour cell invasion and in latest stages as a palliative treatment. Cisplatin is one of the most frequently used and most potent drugs available. It is administered by parenteral route at doses limited by its high and cumulative nephrotoxicity but also by other systemic toxicities (e.g. ototoxicity). Its administration therefore requires many precautions (long hydration procedure, surveillance of the renal function), which mobilize medical personnel. A major limitation of parental CT is the low concentration of drug that successfully reaches the tumour or the metastases. A potential additional modality could be aerosolized CT to localize lung cancer treatment. It has shown a relative local tolerance for cisplatin through preclinical and clinical studies in humans by means of nebulized solutions or liposomal formulations. As a local treatment, aerosolized CT has a clear pharmacokinetic (PK) advantage, as it can increase local exposure while decreasing systemic exposure. However, because CT drugs, such as cisplatin, are active at rather high doses (in the mg range), the duration of administration from nebulizers is very long as it depends on the drug solubility or on drug encapsulation into liposomes. They also pose a high risk of environmental contamination and require HEPA-filtrated hoods during the nebulization procedure. Of all the inhalation devices available to deliver high drug doses, dry powder inhalers (DPIs) were chosen in this work. These were chosen to circumvent the above issues by providing higher deposited doses, in very short timeframes, using a patient-driven device that could help limit environmental exposure to only very low levels of drug. DPI in general also have the advantage of being applicable to both poorly-water-soluble and to water-soluble anticancer drugs. However, because direct deposition of high quantities of anticancer drugs to the lung parenchyma could pose a high risk of local irritation and pulmonary adverse effects, controlled release (CR) of cisplatin from deposited particles in the lung parenchyma was needed. However, in the lungs, foreign undissolved particles are rapidly eliminated by means of naturally occurring clearance mechanisms, in particular macrophage uptake in the alveoli. Therefore, formulation strategies able to limit the particles clearance are needed to assure high lung residence of these CR particles. The formulation strategy of this work was to develop DPI formulation based on solid-lipid microparticles (SLM) able to (i) be deposited into the lung, (ii) control the release of cisplatin and (iii) escape macrophage uptake in order to remain in the lung long enough and at a concentration able to optimize the therapeutic index (i.e. increase the potential therapeutic effect and decrease the potential side effects).The primary objectives of the SLM-based DPI formulations were to (i) exhibit aerodynamic properties compatible with lung cancer patients abilities and cisplatin requirements (e.g. a high deposited fraction, high deagglomeration abilities under low airflow within a low-resistance DPI, deposition in the mg range), (ii) provide a CR matrix for cisplatin in vitro, (iii) be able to be retained into the lung long enough in vivo, (iv) using scalable production techniques and (v) using only potentially well-tolerated excipients.Cisplatin was initially reduced to microcrystals under high-pressure homogenization (HPH) cycles up to 20 000 psi. This procedure permitted uncoated particles with mean diameters below 1.0 μm to be obtained. To assess the cisplatin release abilities of the DPI formulations on the deposited fraction only, a new dissolution test was adapted. This test used a classical paddle apparatus from the pharmacopoeia and a Fast Screening Impactor (FSI). An excipient-free formulation, obtained from the spray dried suspension of cisplatin microcrystals (100% cisplatin) was initially produced. It was compared to a 95:5 cisplatin/tocopheryl polyethylene glycol succinate (TPGS) formulation, which exhibited a higher deposition ability (fine particle fraction (FPF) of 24.2 vs. 51.5% of the nominal dose, respectively). Both exhibited immediate release (IR), with 90% dissolved under 10 minutes.Solid lipid microparticle (SLM)-based formulations were then produced using the cisplatin microcrystalline suspension and various lipid excipients. Those had previously been screened for their ability to be spray dried following their solubilisation in heated isopropanol. The addition of a triglyceride, tristearin (TS), as the main lipid component and if necessary a polyethylene glycol (PEG) excipient-comprising fraction with TPGS or distearoyl phosphoethanolamine polyethylene glycol 2000 (DSPE-mPEG-2000) as a surface modifier, provided spray dried particles with interesting characteristics. These formulations, comprised of at least 50% cisplatin, exhibited high CR abilities in simulated lung fluid at 37°C for more than 24 h (as low as 56% released after 24 h) and a low burst-effect (as low as 24% and 16% after 10 minutes with and without PEGylated excipients, respectively). They also showed high aerodynamic properties, with a high FPF ranging from 37.3 to 50.3% w/w of the nominal dose and a low median mass aerodynamic diameter (MMAD) between 2.0 and 2.4 μm. The process also offered high production yields (> 60%).The best IR DPI formulation (evaluated on the FPF, i.e. cisplatin/TPGS 95:5) and the most promising CR formulations without (i.e. cisplatin/TS 50:50) and with PEGylated excipients (evaluated on CR abilities, i.e. cisplatin/TS/TPGS 50:49.5:0.5) were then administered to CD 1 mice, concurrently to endotracheal nebulization (EN) of a cisplatin solution. This was done using specific endotracheal devices, the Penn-Century Inc. DP-4M© Dry Powder Insufflatorn and for the cisplatin solution, the Microsprayer™ IA-1C©. They were compared to intravenous (IV) injection during a PK study over 48 hours. The administration of DPI formulations required the development of a spray dried diluent (Mannitol:Leucine 10:1) and specific dilution method (3D mixing for 4 hours and double-sieving) to be able to deliver precise and repeatable quantities of powder into the lungs of mice at 1.25 mg/kg dose. A PK study was carried out of the lungs, blood, kidneys, liver, mediastinum and spleen of the mice. The study used a developed and validated electrothermal atomic absorption spectrometry (ETAAS) method. Results showed that endotracheal administration of DPI formulations permitted the exposure of the lungs to cisplatin, expressed as the area under the curve (AUC) to be greatly increased while decreasing the systemic exposure. More precisely, the only formulation that exhibited prolonged lung retention was the one comprising PEGylated excipient (cisplatin/TS/TPGS 50:49.5:0.5), which was observed for ~7 hours. This lung retention was associated with smoother concentration vs. time profiles in blood (higher tmax and lower Cmax), which also confirmed its CR abilities in vivo as dissolved cisplatin is a highly permeable drug. The overall exposure, established by the AUC, helped calculate the target efficiency (Te: the ratio of AUC in the lungs to the sum of AUC in non-target organs) and the target advantage (Ta: ratio of AUC in the lungs by the tested route to the AUC in the lungs by the IV route). For instance, the Ta of the aforementioned formulation (cisplatin/TS/TPGS 50:49.5:0.5) was of 10.9, as compared to 1 for IV, 3.3 for EN, 2.6 for the IR DPI formulation (cisplatin/TPGS 95:5) and 3.7 for the non-PEGylated CR DPI formulation (cisplatin/TS 50:50). In the meantime, the Te for the same formulations were 1.6, 0.09, 1.1, 0.4 and 0.9, respectively, showing again the great efficiency of the inhaled route vs. the IV route in targeting the lungs. More importantly, it showed the added efficiency of the CR DPI formulation with lung retention abilities, provided by the addition of PEGylated excipients. In the last part of the work, maximum tolerated doses (MTD) of formulations were established. These showed that the best candidate, selected based on the PK results (CR DPI with lung retention abilities composed of cisplatin/TS/TPGS 50:49.5:0.5) had better overall tolerance than IR approaches (DPI formulation at cisplatin/TPGS 95:5 and EN of a cisplatin solution). More precisely, it was possible to double the administered dosage for the CR formulation (1.0 mg/kg) vs. the IR DPI and EN (both at 0.5 mg/kg) under a repeated administration scheme (3 times a week for 2 weeks).Moreover, an assessment of the lung tolerance of this best candidate was realized and compared to the IR DPI, EN and the IV route. It was done through analysis of the broncho-alveolar lavage fluid (BALF) 24 hours following a single administration at the pre-determined MTD. IL-1β, IL-6 and TNF-α cytokines were not increased following the administrations. No evidence of tissue damage or cytotoxicity could be observed through quantification of the protein content and of lactate dehydrogenase (LDH) activity. The only observations were a decrease in total cells and an increase in polynuclear neutrophils (PN) cells in the BALF, which was not observed by IV or following the administration of the vehicle of the CR formulation alone (i.e. PEGylated SLM and dry diluent). This increase was not directly linked to the formulation but rather to cisplatin, as it was observed in each cisplatin inhalation experiments, and not with the vehicle of the CR formulation, which was comparable to the non-treated mice.In parallel, we realized a survival study following the administration of the best DPI formulation candidate (cisplatin/TS/TPGS 50:49.5:0.5) vs. the IR DPI candidate (cisplatin/TPGS 95:5), both at their respective MTD under the aforementioned repeated dosing scheme. Cisplatin was administered to mice bearing a grafted orthotopic M109-HiFR lung tumour model, previously developed in the laboratory. The DPI formulations were evaluated against IV administration at each dose (0.5 and 1.0 mg/kg, respectively). This study first confirmed the lower toxicity of the CR approach, as the IR DPI formulation caused a much higher number of deaths during treatment of the grafted mice. The CR formulation administered at 1.0 mg/kg showed a higher survival than the negative control but a tumour response comparable to IV administered at half this dose (0.5 mg/kg). This unexpected outcome with regard to the PK results is explained by the fact that the tumour model is highly metastatic. Mice treated with inhaled formulations died due to distant tumour involvement, while those treated systemically died due to pulmonary tumour involvement. This led us to believe that this kind of treatment may have greater potential in combination, adjuvant to the parenteral route.This work helped establish the proof-of-concept of a cisplatin CR DPI formulation with an up-scalable process. The SLM approach confirmed that encapsulation of drugs exhibiting low solubility, such as cisplatin, was possible using highly hydrophobic excipients and that surface modification was mandatory to provide notable lung retention in vivo. The SLM approach showed good signs of tolerance during the exploratory study but still needs to be confirmed under a chronic scheme using other determinants such as histopathological analyses of the lung tissue. Moreover, comparison of the nephrotoxicity of formulations against that of the IV route should be conducted with appropriate and sensitive methods. Finally, the survival study of the CR DPI formulation showed mitigated results, partly because of the orthotopic model characteristics. This could be proof that inhaled CT has a role to play combined with classical systemic CT. This needs to be assessed in a further study.Le cancer du poumon est le cancer ayant le taux de mortalité le plus élevé au monde, avec un taux de survie global à 5 ans d'environ 15%. Malgré un impact notable des dernières améliorations en matière de prévention, de dépistage, et de classification du cancer du poumon, l'efficacité des traitements classiques n'est toujours pas suffisante et semble avoir atteint un plateau thérapeutique. Ces traitements classiques comprennent de la chirurgie, de la radiothérapie et de la chimiothérapie, le plus souvent en combinaison. La chimiothérapie est utilisée à presque tous les stades: dans les stades opérables et inopérables afin de limiter l'invasion par les cellules tumorales jusqu’aux derniers stades en tant que traitement palliatif. Le cisplatine est l'un des médicaments anticancéreux les plus fréquemment utilisés et les plus puissants actuellement disponibles. Il est administré par voie parentérale à des doses qui sont limitées par sa néphrotoxicité élevée et cumulative mais également par d'autres toxicités systémiques (par exemple, de l'ototoxicité). Son administration nécessite donc de nombreuses précautions (longue procédure d'hydratation, surveillance de la fonction rénale), ce qui mobilise fortement le personnel médical. Une limitation importante de la chimiothérapie parentérale est la faible concentration d’actif qui atteint avec succès la tumeur ou les métastases. Une autre voie d’accès potentielle pourrait être la chimiothérapie inhalée pour traiter le cancer du poumon. Cette approche a montré une relativement bonne tolérance locale pour le cisplatine à travers différentes études précliniques et cliniques chez l'homme au moyen de solutions ou de formulations liposomales nébulisées. En tant que traitement via la voie pulmonaire, la chimiothérapie inhalée présente un avantage pharmacocinétique évident, car elle permet d’augmenter l'exposition locale tout en diminuant l'exposition systémique. Cependant, du fait que les médicaments chimiothérapeutiques, tels que le cisplatine, soient actifs à des doses relativement élevées (dans la gamme du mg), la durée d'administration à partir des nébuliseurs s’avère en pratique très longue car elle dépend principalement de la solubilité de l’actif ou de son encapsulation dans les liposomes. Les nébuliseurs présentent également un risque élevé de contamination de l'environnement et nécessitent de lourds appareillages (hottes filtrantes en particulier) pendant la procédure d’administration.Parmi tous les dispositifs d'inhalation existants, capables de délivrer des doses élevées de médicaments, les inhalateurs de poudre sèche (DPI) semblent être de bons candidats. Ceux-ci ont été choisis dans ce travail afin de contourner les problèmes énumérés ci-dessus, en fournissant des doses pulmonaires plus élevées, dans des délais très courts. De plus, ces dispositifs sont activés par le flux inspiratoire du patient, ce qui pourrait aider à limiter l'exposition environnementale à des niveaux très faibles. Les inhalateurs à poudre sèche présentent également l'avantage d'être utilisables à la fois avec des médicaments solubles et des médicaments peu solubles dans l’eau. Malgré tout, étant donné que la déposition directe de quantités élevées de médicaments chimiothérapeutiques dans le parenchyme pulmonaire pourrait présenter un risque élevé d'irritation et d'effets indésirables locaux, une libération contrôlée du cisplatine à partir de particules déposées dans le parenchyme pulmonaire s’avère nécessaire. Cependant, dans les poumons, ces particules non dissoutes d’origine étrangère sont rapidement éliminées par les mécanismes d’élimination, en particulier par la clairance par les macrophages au niveau des alvéoles. Par conséquent, des stratégies de formulation capables de limiter la clairance des particules sont nécessaires pour assurer une résidence pulmonaire élevée de ces particules à libération contrôlée.La stratégie de formulation de ce travail a donc consisté à développer une formulation pour inhalateur à poudre sèche à base de microparticules lipidiques solides capable de (i) être déposées dans le poumon, (ii) de contrôler la libération du cisplatine et (iii) de rester dans le poumon suffisamment longtemps dans le but d’optimiser l'indice thérapeutique (c'est-à-dire augmenter le potentiel thérapeutique du cisplatine et diminuer ses potentiels effets secondaires).Les objectifs principaux des formulations basées sur les microparticules lipidiques solides étaient (i) de présenter des hautes charges en cisplatine au sein des microparticules lipidiques tout en présentant des propriétés aérodynamiques compatibles avec la capacité pulmonaire des patients atteints de cancer du poumon (par exemple, une fraction déposée élevée et une capacité élevée à la désagglomération sous faible débit d'air dans un inhalateur de faible résistance), (ii) de fournir une matrice capable de libérer le cisplatine de manière contrôlée in vitro, (iii) d’être capable de rester dans le poumon suffisamment longtemps in vivo, tout cela (iv) en utilisant des techniques de production ayant une bonne capacité d’augmentation d’échelle et (v) de n’utiliser que des excipients potentiellement bien tolérés au niveau du poumon.Le cisplatine a été initialement réduit sous forme microcristalline à l’aide de cycles d'homogénéisation à haute pression jusqu'à 20 000 psi. Cette procédure a permis d'obtenir des particules non enrobées ayant un diamètre moyen inférieur à 1.0 μm. Afin d’évaluer les capacités de libération du cisplatine des formulations à partir de la fraction capable théoriquement de se déposer dans les poumons, un nouveau test de dissolution a été adapté à partir d’un appareil à palettes classique de la pharmacopée et d’un impacteur à cascade « Fast Screening Impactor ». Une formulation sans excipient, obtenue à partir de la suspension de cisplatine, soumise à la technique de séchage par l’atomisation (100% de cisplatine) a été produite comme point de départ. Celle-ci a ensuite été comparée à une formulation de cisplatine/tocophéryl polyéthylène glycol succinate (TPGS) (95:5), qui présentait une capacité de déposition pulmonaire in vitro (fraction de particules fines (FPF) de 24.2% pour la première et de 51.5% pour la deuxième, exprimée par rapport à la dose nominale). Toutes deux ont démontré des capacités de libération immédiate, avec 90% du cisplatin dissous en moins de 10 minutes.D’autres formulations, cette fois élaborées sous la forme de microparticules lipidiques solides ont ensuite été produites à partir de la suspension microcristalline de cisplatine et de divers excipients lipidiques. Ces microparticules avaient préalablement été testées pour leur aptitude à être séchées par atomisation après solubilisation des excipients dans de l'isopropanol chaud. L’ajout d’un triglycéride, la tristéarine (TS), comme excipient lipidique principal et également d’une fraction comprenant un excipient contenant du polyéthylène glycol (PEG), à l’aide de TPGS ou de distéaroyl phosphoéthanolamine polyéthylène glycol 2000 (DSPE-mPEG-2000) a montré des résultats intéressants. Ces formulations, ayant une teneur en cisplatine d’au moins 50%, ont présenté des aptitudes élevées pour la libération contrôlée dans le fluide pulmonaire simulé in vitro à 37 °C, et ce, pendant plus de 24 h (jusqu'à 56% libérées après 24 h) ainsi qu’un faible « burst-effect » (de seulement 24% et 16% après 10 minutes avec et sans excipients PEGylés, respectivement). Elles ont également montré des propriétés aérodynamiques élevées, avec une FPF élevée allant de 37.3 à 50.3% m/m par rapport à la dose nominale et un diamètre aérodynamique compris entre 2.0 et 2.4 μm. Le meilleur candidat à libération immédiate (évaluée sur base de la FPF, soit la formulation cisplatine/TPGS 95:5 m/m) et les formulations à libération contrôlée les plus prometteuses n’incluant pas d’excipients PEGylés (cisplatine/TS 50:50 m/m) et incluant des excipients PEGylés (évalués sur les capacités de libération contrôlée, c'est-à-dire la formulation cisplatin/TS/TPGS 50:49.5:0.5 m/m/m) ont ensuite été administrées à des souris CD-1, en comparaison d’une nébulisation endotrachéale d'une solution de cisplatine. Ceci a été fait à l’aide de dispositifs endotrachéaux dédiés aux poudres pour le DP-4M© « Dry Powder Insufflator » et aux solutions pour le Microsprayer™ IA-1C© de Penn-Century. Ces formulations ont été comparées à l'injection intraveineuse (IV) au cours d’une étude pharmacocinétique étendue sur 48 heures.L'administration de formulations de poudres sèches pour inhalation a nécessité le développement préalable d'un diluant par atomisation (Mannitol:Leucine 10:1 m/m) ainsi que d’une méthode de dilution des poudres (mélange tridimensionnel pendant 4 heures et suivi d’un double-tamisage) afin de pouvoir délivrer des quantités précises et répétables de poudre dans les poumons de souris à la dose d’1.25 mg/kg. Le suivi des paramètres pharmacocinétiques a ainsi pu être réalisé au niveau des poumons, du sang, des reins, du foie, du médiastin et de la rate des souris. Ceci a été fait à l’aide d’une méthode de spectrométrie d'absorption atomique électrothermique, qui a été préalablement développée et validée. Les résultats obtenus ont montré que l'administration endotrachéale de formulations de poudres sèches permettait d’augmenter fortement l'exposition des poumons par le cisplatine, exprimée en aire sous la courbe (AUC) tout en diminuant l'exposition systémique. Plus précisément, la seule formulation présentant une rétention pulmonaire prolongée était celle qui comprenait un excipient PEGylé (cisplatine/TS/TPGS 50:49.5:0.5 m/m/m), ce qui a été observé pendant environ 7 heures. Cette rétention pulmonaire a été associée à des profils de concentration en fonction du temps plus réguliers dans le sang (tmax supérieur et Cmax inférieur), ce qui a également confirmé ses capacités de libération contrôlée in vivo car la perméabilité de l’épithélium pulmonaire pour le cisplatine dissous s’est avérée très élevée. L'exposition globale établie à partir de l’AUC a permis de calculer l’efficacité de ciblage (Te: rapport de l'AUC mesurée dans les poumons et de la somme des AUC mesurées dans les organes non cibles) et l’avantage du ciblage (Ta: rapport de l’AUC mesuré dans les poumons suite à l’administration pulmonaire et de l'AUC mesurée dans les poumons suite à l’administration par la voie IV). Par exemple, le Ta de la formulation décrite ci-dessus (cisplatine/TS/TPGS 50:49.5:0.5 m/m/m) était de 10.9, comparativement à 1 pour l’IV, 3.3 pour la nébulisation endotrachéale, 2.6 pour la formulation de poudre sèche à libération immédiate (cisplatine/TPGS 95:5 w/w) et 3.7 pour la formulation de poudre sèche à libération contrôlée ne comprenant pas d’excipient PEGylé (cisplatine/TS 50:50). Dans le même temps, le Te mesuré pour les mêmes formulations était de 1.6, 0.09, 1.1, 0.4 et 0.9, respectivement, démontrant également le rendement élevé de la voie inhalée par rapport à la voie IV dans sa capacité à cibler les poumons. Plus important encore, ceci a démontré le grand avantage des capacités de rétention pulmonaire de la formulation à libération contrôlée comprenant un excipient PEGylé.Dans la dernière partie de ce travail, les doses maximales tolérées (DMT) des formulations ont été déterminées. Le meilleur candidat, choisi en fonction des résultats de pharmacocinétique (formulation à libération contrôlée ayant des capacités de rétention pulmonaire composé de cisplatine/TS/TPGS 50:49.5:0.5 m/m/m), avait une meilleure tolérance globale que les deux approches à libération immédiate testées (formulation de poudre sèche cisplatine/TPGS 95:5 et la nébulisation endotrachéale d'une solution de cisplatine). Plus précisément, il s’est avéré possible de doubler le dosage administré pour la formulation à libération contrôlée (1.0 mg/kg) par rapport à la poudre sèche à libération immédiate et à la nébulisation endotrachéale (toutes les deux à 0.5 mg/kg) suivant un schéma d'administration chronique (3 fois par semaine pendant 2 semaines). De plus, une évaluation de la tolérance pulmonaire de cette formulation à libération prolongée a été réalisée et comparée à la poudre sèche à libération immédiate, à la nébulisation endotrachéale et à la voie IV. Elle a été réalisée par analyse du liquide provenant du lavage broncho-alvéolaire, 24 heures après une administration unique à la dose maximale tolérée préalablement déterminée pour chaque formulation. Aucune augmentation des cytokines IL-1β, IL-6 et TNF-α n’a pu être détectée à la suite des administrations. Aucunes preuves de lésion tissulaire ou de cytotoxicité n'ont pu être observées au travers du dosage de la teneur en protéines totale et de l'activité de la lactate déshydrogénase. Les seules observations qui ont pu être faites ont été une diminution des cellules totales et une augmentation des polynucléaires neutrophiles dans le lavage broncho-alvéolaire, ce qui n'a pas été observé suite à l’administration IV ou après l'administration du véhicule de la formulation à libération contrôlée seul (c'est-à-dire les microparticules lipidiques solides PEGylées et le diluant). Cette augmentation ne semble pas liée aux microparticules lipidiques solides ou au diluent mais probablement à l’exposition pulmonaire au cisplatine, car cette augmentation a été observée pour chaque groupe inhalé contenant du cisplatine. Le cisplatine a ensuite été administré à des souris qui ont été greffées de manière orthotopique par une lignée murine de carcinome pulmonaire M109-HiFR, modèle préclinique préalablement développé au sein de notre laboratoire. Les formulations de poudres sèches ont été évaluées par rapport à l'administration IV à chaque dose testée (0.5 et 1.0 mg/kg, respectivement). Cette étude a d'abord confirmé la toxicité plus faible de l'approche à libération contrôlée, car la formulation à libération immédiate a causé un nombre beaucoup plus élevé de décès pendant le traitement des souris greffées. La formulation à libération contrôlée administrée à 1.0 mg/kg, a montré une survie plus élevée que le contrôle négatif, mais une réponse comparable à la dose IV administrée à la moitié de la dose (0.5 mg/kg). Ce résultat inattendu par rapport aux résultats de l’étude pharmacocinétique s'explique probablement par le fait que le modèle de tumeur utilisé est hautement métastatique. Les souris traitées avec des formulations inhalées sont mortes en raison de tumeurs secondaires distantes par rapport à la tumeur primaire implantée au niveau du poumon, alors que celles traitées par la voie systémique sont mortes en raison d’un envahissement tumoral pulmonaire. Cela nous amène à penser que ce type de traitement inhalé pourrait avoir un plus grand potentiel en combinaison à la voie parentérale. Ce travail a ainsi permis d’établir la preuve du concept de formulation à base de poudre sèche de cisplatine à libération contrôlée, en utilisant un processus de fabrication capable de subir une mise à l’échelle industrielle. L’utilisation de microparticules lipidiques solides a confirmé que l'encapsulation d’actifs présentant une certaine hydrophilie, comme le cisplatine, était possible en utilisant des excipients hautement hydrophobes et qu'une modification de leur surface était cependant obligatoire pour obtenir une rétention pulmonaire intéressante in vivo. Les microparticules lipidiques solides ont montré de bons signes de tolérance au cours de l'étude exploratoire, mais celle-ci doit encore être confirmée avec une administration chronique des poudres. Ceci doit être fait en suivant des paramètres supplémentaires, tels que des analyses histologiques du tissu pulmonaire. De plus, la comparaison de la néphrotoxicité des formulations avec celle mesurée par la voie IV doit être effectuée avec des méthodes appropriées et sensibles. Enfin, l'étude de survie de la formulation à libération prolongée a montré des résultats mitigés, en partie à cause des caractéristiques du modèle orthotopique de tumeur pulmonaire. Cependant, il semblerait que la chimiothérapie inhalée à un rôle important à jouer en combinaison avec la chimiothérapie systémique classique. Ceci doit être évalué dans une étude future.
Doctorat en Sciences biomédicales et pharmaceutiques (Pharmacie)
info:eu-repo/semantics/nonPublished

Tese de mestrado, Engenharia Farmacêutica, Universidade de Lisboa, Faculdade de Farmácia, 2018
The Pharmaceutical industry is increasingly investing in biopharmaceuticals. Moreover, alternative delivery routes are being investigated over the conventional parenteral administration route, which presents several inconveniences for the patients including discomfort, pain and the impossibility of self-administration in the majority of cases. Thus, pulmonary and nasal delivery are emerging as alternative administration routes for the treatment of systemic diseases and administration of biopharmaceuticals, both allowing a rapid absorption and onset of therapeutic action. Additionally, nasal route allows nose-to-brain delivery through the olfactory neuroepithelium. The present thesis is divided into two main studies. The first one aimed at the optimization of a Spray Drying (SD) Process using an Ultrasonic Nozzle (USN) 1 to produce dry powder inhalers (DPI) with a particle size between 1 and 5 μm. Although the physical characterization was within the expected for a DPI formulation of excipient A:excipient B (4:1), the aim of this study was not completely achieved since to obtain a powder with a particle size within the inhalable size range, the SD process would require extremely diluted feed solutions, leading to long run times. Thus, the optimization of the SD process was not possible. The second study aimed at the assessment of a SD process with a Two-Fluid Nozzle (TFN) and a USN 3 to produce nasal dry powder formulations with a small molecule as a model Active Principle Ingredient (API) and, ultimately, with an API with medical application through nasal route. A design of experiments was performed for each nozzle to assess which one proves to be the best candidate to produce dry powders for nasal delivery. The excipients chosen, excipient A and excipient C, proved to be suitable for dry nasal delivery with potential compatibility with biopharmaceuticals allowing its future delivery though this route. All powders were amorphous and had a particle size within the nasal size range of 10 – 45 μm. The aerodynamic performance showed a correlation between the powders particle size distribution (PSD) and the fraction retained in the nasal cavity. When comparing powders with the same volumetric median particle size (Dv50), the powder with a narrower PSD (lower value of span) was the one that presented a lower percentage of powder retained in the gastrointestinal tract and respiratory system. Thus, although the TFN could be used at a first screening phase, the USN 3 is the preferable atomizing system since it produces powders with a significant narrower PSD therefore maximizing deposition on the nasal area and minimizing the fraction of powder with potential to reach the deep lungs. Ultimately, this thesis showed the importance of the technology selected to produce dry powder formulations intended for nasal delivery since it could have impact on the deposition profile of the powders.
Atualmente, a indústria Farmacêutica tem vindo a investir cada vez mais na investigação e produção de biofármacos. Estas macromoléculas são, na maioria dos casos, administradas por via parenteral (intravenosa, intramuscular ou subcutânea). Contudo, têm vindo a ser investigadas vias de administração alternativas à parenteral pois esta apresenta vários inconvenientes para os pacientes, nomeadamente desconforto, dor e impossibilidade de autoadministração na maioria dos casos. Assim, vias de administração como a pulmonar e nasal surgem como alternativas para o tratamento de doenças sistémicas e para administração de biofármacos, permitindo uma rápida absorção e início da ação terapêutica. Adicionalmente, a via nasal permite a administração direta de fármacos para o sistema nervoso central através do neuroepitélio olfativo. A presente tese encontra-se dividida em dois estudos. O primeiro visou a otimização de um processo de secagem por atomização usando um atomizador ultrassónico 1 (USN) para produzir inaladores de pó seco (DPI) com um tamanho de partícula adequado para inalação, entre 1 e 5 μm. Embora para uma formulação de DPI de excipiente A:excipiente B (4:1) a caracterização física estivesse dentro do esperado, o objetivo deste estudo não foi completamente atingido uma vez que não foi possível otimizar o processo de secagem por atomização. Para obter um pó com tamanho de partícula dentro da gama de inalação, o processo de secagem exigiria soluções de alimentação extremamente diluídas, levando a experiências extremamente longas. Por outro lado, este processo apresentou baixa rentabilidade. Estas foram as principais razões pelas quais a presente tese mudou o foco da otimização de um processo de secagem por atomização visando a produção de inaladores de pó seco para administração pulmonar para a otimização de um processo para produção de pós para administração nasal. O segundo estudo teve como objetivo a comparação de dois sistemas de atomização (atomizador de dois fluidos e atomizador ultrassónico 3) para produzir formulações nasais de pó seco com uma molécula de síntese química modelo como princípio ativo e, numa última fase, com um princípio ativo com aplicação médica por via nasal. Para o atomizador de dois fluidos, de forma a comparar as propriedades dos pós obtidos, foi realizado um desenho de experiências onde os parâmetros variados foram a composição da formulação e o caudal de atomização. Para as variáveis, dois níveis foram estabelecidos. Dentro da formulação, a concentração de princípio ativo Y foi mantida constante a 0,01 %, o excipiente A variou entre 20,00 e 50,00% e o excipiente C entre 49,99 e 79,99 %. O nível mínimo de excipiente A foi definido com base na concentração mínima necessária para estabilizar a Cu, Zn-superóxido dismutase. Adicionalmente, a menor concentração do excipiente C foi estabelecida com base na suposição de que o aumento do tempo de residência da formulação na cavidade nasal é particularmente favorável quando a administração de macromoléculas é necessária. Em termos de parâmetros do processo, o caudal de atomização (Fatom) variou entre 0,27 × Fatom_ref e 0,43 × Fatom_ref, sendo que os restantes parâmetros foram mantidos constantes nas cinco condições estudadas. Um estudo com o atomizador ultrassónico 2 foi realizado visando uma primeira comparação entre as propriedades de pós produzidos por um atomizador de dois fluidos e um atomizador ultrassónico. A formulação escolhida apresentou o princípio ativo Y numa concentração de 0,01 %, o excipiente A a 20,00 % e o excipiente C a 79,99 %. Apesar do pó produzido ter apresentado um Dv50 fora da gama para administração nasal (47 μm), foi possível concluir que este tipo de atomizador produz pós com uma distribuição de tamanho de partículas mais estreita em comparação com o atomizador de dois fluidos, sendo a principal vantagem deste sistema de atomização. Para o atomizador ultrassónico 3, semelhante ao estudo realizado com o atomizador de dois fluidos, foi realizado um desenho de experiências onde os parâmetros variados foram a composição da formulação e o caudal de alimentação, de modo a comparar as propriedades dos pós obtidos e, em última análise, estabelecer o melhor sistema de atomização para a produção de pós para administração nasal. Para as variáveis, dois níveis foram estabelecidos. Dentro da formulação, a concentração do princípio ativo Y foi mantida constante a 0,01%, o excipiente A variou entre 20,00 - 50,00% e o excipiente C entre 49,99 - 79,99%. Em termos de parâmetros do processo, o caudal de alimentação (Ffeed) variou entre 0,53 × Ffeed_ref e 0,67 × Ffeed_ref, sendo que os restantes parâmetros foram mantidos constantes nas cinco condições estudadas. Todos os pós produzidos estavam num estado amorfo e apresentavam um tamanho de partícula dentro da gama para administração nasal, entre 10 a 45 μm. Comparando os pós produzidos com o atomizador de dois fluidos e com o atomizador ultrassónico 3, podemos concluir que os primeiros apresentaram uma diferença significativa no tamanho médio volumétrico de partícula (Dv50) consoante o valor estabelecido de caudal de atomização. Assim, quando o caudal de atomização foi maior (0,43 × Fatom_ref), o Dv50 foi menor (11 μm) e vice-versa. Contudo, o mesmo não foi observado nos pós produzidos com o atomizador ultrassónico 3, uma vez que o Dv50 apenas variou entre 37,1 e 40,4 μm para a gama de caudal de alimentação estabelecida (0,53 × Ffeed_ref a 0,67 × Ffeed_ref), sugerindo que a mesma deveria ter sido maior, de forma a que houvessem diferenças significativas no Dv50. O desempenho aerodinâmico in-vitro dos pós foi avaliado utilizando um Impactador em cascata de Andersen reduzido equipado com uma câmara de expansão de vidro de 5 L. Apenas três estágios foram utilizados: 0, 2 e F. O estágio 0 retém as partículas com tamanho de partícula aerodinâmico superior a 9,0 μm, enquanto que os estágios 2 e F retêm partículas com um tamanho de partícula aerodinâmico entre 4,7 a 9,0 μm e 0,4 a 4,7 μm, especificando a fração da dose emitida que pode ser retida nas passagens intranasais, no trato gastrointestinal e nos pulmões, respetivamente. O desempenho aerodinâmico mostrou uma correlação entre a distribuição do tamanho de partículas dos pós e a fração retida na cavidade nasal (R2 = 0,982; Q2 = 0,965). Adicionalmente, o valor do span foi considerado o fator crítico para este desempenho, na medida em que comparando pós com o mesmo Dv50, quanto mais estreita a distribuição do tamanho de partículas, menor a fração que atinge o trato gastrointestinal e o sistema respiratório. Assim, embora o atomizador de dois fluidos possa ser utilizado numa primeira fase de desenvolvimento, o atomizador ultrassónico 3 é o sistema de atomização preferível, pois produz pós com uma distribuição de tamanho de partículas mais estreita, maximizando a deposição na área nasal e minimizando a fração de pó com potencial para atingir os pulmões. Tendo como objetivo a produção de um pó com relevância médica para administração nasal (princípio ativo Z, indicado no tratamento de enxaquecas agudas), um último estudo foi realizado nas melhores condições de processo obtidas com o atomizador de dois fluidos. A formulação apresentou 1,0 % de princípio ativo Z e 99,0 % de excipiente C como agente mucoadesivo. O pó obtido apresentou-se num estado amorfo, tendo um Dv50 de 20 μm. Em termos de performance aerodinâmica, o pó apresentou uma fração retida no estágio 0 (cavidade nasal) de 92,0% e uma fração inferior a 3,0% nos estágios 2 e F (trato gastrointestinal e pulmões, respetivamente). Por fim, esta tese mostrou a importância da tecnologia selecionada para produzir formulações de pó seco destinadas a administração nasal, uma vez que a mesma poderá ter impacto no perfil de deposição dos pós produzidos.
Hovione FarmaCiência

The aim of this research was to develop and fully investigate a novel method of antibiotic drug delivery to the lung that will address problems with current therapeutic regimens for treatment of airway infections. To demonstrate the performance of prepared formulations, the design of suitable characterization methods were also aimed. A novel dissolution method for evaluating the in vitro dissolution behavior of inhalation formulations was developed. The membrane holder was designed to enclose previously air-classified formulations so that they could be uniformly tested in the dissolution apparatus. Dissolution procedures, the apparatus, the dose collection, the medium, and test conditions were developed and the dissolution behaviors of test compounds were evaluated by experimental and mathematical analysis. It was proved that the aerodynamic separation of formulation prior to dissolution assessment have a significant influence on the dissolution profiles. The optimized test method using the membrane holder was applied to evaluate in vitro dissolution profiles of the manufactured formulations of rifampicin (RF). The carrier/excipient-free RF dry powder formulation was investigated. The rifampicin dihydrate (RFDH) powders having MMAD of 2.2 um were prepared using a simple recrystallization process. The RFDH powders have a thin flaky structure, and this unique morphology provides improved aerosolization properties at maximal API loading. The manufactured RFDH formulation showed 80% drug release within 2 hours. To retard the release rate of RF, the prepared RFDH crystals were coated with hydrophobic polymer, PLA or PLGA, using spray-dryer equipped with multi-channel spray nozzles. The multi-channel spray nozzle used in this study has two separate nozzles for aqueous solution and one for gas fluid. The RFDH crystals and the coating solutions were sprayed through the two separate liquid nozzles at the same time. The coated RFDH formulations were prepared using multi-channel spray nozzles. The coated formulations contained at least 50% w/w of RF with no change of their flaky morphology. The initial RF release was lowered by coating; the lowest initial RF release was observed from the coated powders with PLA polymer as 32% among the coated formulations. Overall, the 80% of RF was released within 8 hours. The RFDH and coated RFDH formulations delivered via the pulmonary route would be anticipated to provide higher local (lung) drug concentrations than that of orally delivered powders. Particularly, the coated RFDH powders deposited in the alveolar region may prolong the drug residence time in the site of infections. Additionally, it was proved that the RFDH and coated RFDH formulations provided much better stability than the amorphous RF.
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碩士
大同大學
生物工程學系(所)
97
The respiratory tract is one of the oldest routes for the administration of drugs more than 4000 years ago. Treatment of asthma by inhaled sympathetic agents began in earnest in the 1950s, and now such ‘topical’ or targeted treatment with inhaled drugs is considered for treating many other lung diseases. The drugs delivery by inhalation can be delivered with very rapid action, low metabolism and high bioavailability; and decreased systemic effects than intravenous injection. Dry powder inhaler formulation and delivery devices development and improvement in recently. But Patients with respiratory failure are often unable to inhale powdered aerosol medications such as long-acting anticholinergic agent- Tiotropium, which are important treatments for chronic obstructive pulmonary disease and asthma. With a proper interface, powdered long-acting agonists and long-acting anticholinergics can be easily delivered via tracheostomy tube. It is important to improved continuity and quality of care. This experiment selected a proper interface to evaluated effect of simulating dry powder inhaler emitted through tracheostomy tube. We estimates may the powdered long-acting anticholinergics can deliveried to the lungs.

Dry Powder inhalers (DPIs) have been one of the most promising developments in pulmonary drug delivery systems. In general, DPIs are more effective than systemic administrations and convenient to use. However, delivering high-dose antibiotics through a DPI is still a challenge because high powder load may need a very large inhaler or increase the incidence of local adverse effects. Spray drying has been increasingly applied to produce DPI formulations for high-dose antibiotics; nevertheless, many spray-dried particles are amorphous and physically unstable during storage, particularly under the humid environment.

My research focuses on addressing critical challenges in physical stability of DPIs for spray-dried high-dose antibiotics. The effects of moisture-induced crystallization on physical stability and aerosol performance of spray-dried amorphous Ciprofloxacin DPI formulations stored at different humidity conditions were studied. Our study not only provided a mechanistic understanding in the impact of crystallization on aerosol performance but also developed novel approaches for improving stability of spray-dried formulations used in DPI.

Our work has shown that recrystallization of amorphous spray-dried Ciprofloxacin led to significant changes in aerosol performance of DPIs upon storage, which cause critical quality and safety concerns. These challenges have been solved through co-spray-drying Ciprofloxacin with either excipient such as leucine or synergistic antibiotic like Colistin. Co-spray-drying Ciprofloxacin with Colistin not only improved physical and aerosol stability but also enhanced antibacterial activity which is a great advantage for treating ‘difficult to cure’ respiratory infections caused by multidrug resistant bacteria.

My research work is a sincere effort to maximize the utility and efficacy of high-dose DPI, an effective delivery tool for treating severe resistant bacterial respiratory infections.

Tese de mestrado, Engenharia Farmacêutica, Universidade de Lisboa, Faculdade de Farmácia, 2014
The present work describes the determination of the influence of specific raw material attributes on a drug product for inhalation performance. Each raw material (drug substance and sugar-based excipients) was characterized in terms of properties that can have an impact in the drug product formulation and performance, such as particle size distribution and dynamic powder properties. The formulations’ performance was determined through its aerodynamic particle size distribution (aPSD) by using a Next Generation Impactor. With this work the relationship between attributes such as the content of fine excipient (or ternary agent), the particle size distribution of each raw material, the powder’s flow energy, the surface of the drug substance (and hence the type of technology used for particle production and/or particle size reduction) was studied and determined to be of key importance. It was also found that these attributes have an influence in the drug product’s performance and so it becomes important to understand them in order to obtain product quality. Identifying and understanding the critical raw materials and their attributes’ relationship with each other and with process parameters will enable formulation scientists to better understand the process and design a quality product and manufacturing process that generate consistent quality product fit for purpose.
No presente estudo descreve-se a determinação da influência dos atributos das matérias-primas na performance de produtos formulados de inalação. Cada matéria-prima (substância ativa e excipientes) foi caracterizada relativamente a propriedades que poderão ter impacto na formulação de produtos finais e sua performance, tais como a distribuição do tamanho de partícula e propriedades de fluidização dos pós. A performance de cada formulação estudada foi determinada por distribuição do tamanho aerodinâmico de partícula (aPSD) através da utilização de um impactador em cascata, Next Generation Impactor. Com este estudo foi possível demonstrar que o conteúdo em excipientes de tamanho pequeno de partícula (ou agentes terciários), a distribuição do tamanho das partículas, a energia de fluidização do pó, o tipo de superfície do princípio ativo (e portanto o tipo de tecnologia utilizado para produção e/ou redução da partícula de princípio ativo) são parâmetros de extrema importância, que ajudam a compreender como a qualidade do produto final pode ser afetada. A identificação e compreensão das matérias-primas críticas e dos seus atributos, da relação que estas estabelecem entre si e com os parâmetros de processo, permitem aos formuladores perceber o processo e desenhá-lo de modo a que este produza consistentemente um produto de qualidade para o fim desejado.

Yes
Two different types of dry powder inhalers (Easyhaler® and RS01®) were used in this work to evaluate the ex vivo and in vitro performance of a budesonide inhaled formulation with recrystallised mannitol, commercial DPI-grade mannitol, or lactose. The aerodynamic performance of the budesonide formulation with recrystallised mannitol was superior when RS01® was used (FPF = 45.8%) compared to Easyhaler® (FPF = 14%). However, the aerodynamic profile was very poor in both devices when commercial mannitol was used. Interestingly, the aerosol performance of the marketed budesonide formulation significantly improved when RS01® was used compared to Easyhaler® (the original device for the formulation). Due to the significant increases in the surface energy of the commercial mannitol formulation, the aerodynamic performance of the formulation was very poor. This work demonstrates the impact of inhaler devices on the performance of inhaled formulations and considers the particle surface energy during formulation development.
The full-text of this article will be released for public view at the end of the publisher embargo on 14 Jun 2021.

No
Purpose The relative lung bioavailability of salbutamol sulfate particles produced using supercritical fluids (SEDS¿) and delivered by dry powder inhaler (DPI) was compared with the performance of a conventional micronized drug DPI using the same device design (Clickhaler¿, Innovata Biomed). Materials and Methods Twelve healthy volunteers and 11 mild asthmatic patients completed separate four-way randomised cross-over studies, assessing the relative bioavailability of salbutamol sulfate (urinary excretion method), formulated as SEDS¿ particles (three batches) and micronized particles (Asmasal¿ inhaler, UCB Pharma Ltd). Post-treatment improvements in patient lung function were assessed by measuring FEV1. Physicochemical evaluation of the three SEDS¿ batches revealed inter-batch differences in particle size and shape. Results There was no significant difference in the relative lung bioavailability of salbutamol and its bronchodilator response between the best performing SEDS¿ formulation and the Asmasal¿ inhaler in volunteers and patients, respectively. SEDS¿ salbutamol sulfate showing wafer like morphology gave greater fine particle dose, relative lung bioavailability and enhanced bronchodilation compared to other SEDS¿ batches containing elongated particles. Conclusions Active Pharmaceutical Ingredient (API) manufactured using supercritical fluids and delivered by DPI can provide similar lung bioavailability and clinical effect to the conventional micronized commercial product. Product performance is however notably influenced by inter-batch differences in particle characteristics.

Trabalho Final de Mestrado Integrado, Ciências Farmacêuticas, Universidade de Lisboa, Faculdade de Farmácia, 2019
O ser humano utiliza diversas vias para administração de fármacos para tratamentos de diversas patologias. Umas dessas vias é a via pulmonar, a qual o Homem já utiliza há milhares de anos, através da inalação de substâncias que vão exercer ação farmacológica no organismo. Um dos tipos de dispositivos que foram desenvolvidos são os Dry Powder Inhaler (DPI’s), ou seja inaladores de pó seco. Mas o fármaco utilizado neste tipo de dispositivos necessita de excipientes que permitirão uma melhor administração, resultando num efeito terapêutico mais eficiente. Estes excipientes têm uma função importante pois ditam se o fármaco é administrado eficazmente, o que influencia o seu efeito terapêutico. Nesta monografia será abordada a temática dos excipientes utilizados em DPI’s, mais concretamente, lactose, manitol, glucose monohidratada, sorbitol, trealose, eritritol, ciclodextrinas, PLGA e hidroxiapatita.
Humans use various routes for drug administration to treat various pathologies. One of these pathways is the pulmonary pathway, which humans have been using for thousands of years, through the inhalation of substances that will exert pharmacological action in the body. One of the types of devices that have been developed are Dry Powder Inhaler (DPI's), ie dry powder inhalers. But the drug used in such devices needs excipients that will allow better administration, resulting in a more efficient therapeutic effect. These excipients play an important role as they dictate whether the drug is administered effectively, which influences its therapeutic effect. This monograph will address the topic of excipients used in DPIs, namely lactose, mannitol, glucose monohydrate, sorbitol, trehalose, erythritol, cyclodextrins, PLGA and hydroxyapatite.

Advancements in pulmonary drug delivery technologies have improved the use of dry powder inhalation therapy to treat respiratory and systemic diseases. Despite remarkable improvements in the development of dry powder inhaler devices (DPIs) and formulations in the last few years, an optimized DPI system has yet to be developed. In this work, we hypothesize that Thin Film Freezing (TFF) is a suitable technology to improve inhalation therapies to treat lung and systemic malignancies due to its ability to produce brittle powder with optimal aerodynamic properties. Also, we developed a performance verification test (PVT) for the Next Generation Cascade Impactor (NGI), which is one of the most important in vitro characterization methods to test inhalation. In the first study, we used TFF technology to produce amorphous and brittle particles of rapamycin, and compared the in vivo behavior by the pharmacokinetic profiles, to its crystalline counterpart when delivered to the lungs of rats via inhalation. It was found that TFF rapamycin presented higher in vivo systemic bioavailability than the crystalline formulation. Subsequently, we investigated the use of TFF technology to produce triple fixed dose therapy using formoterol fumarate, tiotropium bromide and budesonide as therapeutic drugs. We investigated applications of this technology to powder properties and in vitro aerosol performance with respect to single and combination therapy. As a result, the brittle TFF powders presented superior properties than the physical mixture of micronized crystalline powders, such as excellent particle distribution homogeneity after in vitro aerosolization. Lastly, we developed a PVT for the NGI that may be applicable to other cascade impactors, by investigating the use of a standardized pressurized metered dose inhaler (pMDI) with the NGI. Two standardized formulations were developed. Formulations were analyzed for repeatability and robustness, and found not to demonstrate significant differences in plate deposition using a single NGI apparatus. Variable conditions were introduced to the NGI to mimic operator and equipment failure. Introduction of the variable conditions to the NGI was found to significantly adjust the deposition patterns of the standardized formulations, suggesting that their use as a PVT could be useful and that further investigation is warranted.
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Dissertação de Mestrado em Biotecnologia Farmacêutica apresentada à Faculdade de Farmácia
O presente trabalho foi baseado no desenvolvimento de um método analítico utilizando a abordagem de Analytical Quality by Design (AQbD). Esta abordagem é utilizada quando existe a necessidade de se obter um conhecimento completo do método e, ao mesmo tempo, controlar os possíveis fatores que podem influenciar os resultados no trabalho de rotina. O método analítico escolhido foi o Next Generation Impactor (NGI) que tem a capacidade de avaliar a distribuição aerodinâmica do tamanho de partícula de um aerossol. Este método foi escolhido com o objetivo de melhorar a sua robustez e, ao mesmo tempo, criar um fluxo de trabalho representativo das etapas essenciais no seu desenvolvimento.A parte experimental começou após a realização de uma análise de risco do método geral de NGI usando uma ferramenta chamada “Failure Mode and Effect Analysis”. Com todas as possíveis variáveis críticas identificadas, foi desenvolvido um método de NGI e foram definidas as condições ótimas para cada variável.As variáveis testadas foram: o número de cápsulas utilizadas em cada teste, tempo e tipo de agitação de cada componente, influência do revestimento e presença de perdas através do interstage. Ao longo de todos os testes, verificou-se que o balanço de massa esteve sempre dentro dos critérios definidos pela Food and Drug Administration exceto no teste realizado sem revestimento. As variáveis que tiveram influência considerável nos resultados obtidos estão relacionadas com a aplicação da solução de revestimento e com o tempo de agitação durante a recuperação. Testes adicionais foram realizados para avaliar a estabilidade de padrões e amostras.No final do trabalho, concluiu-se que a abordagem AQbD no desenvolvimento do NGI foi muito útil, uma vez que permitiu avaliar quais as variáveis críticas do método com maior impacto nos resultados obtidos.
The present work was based on the development of an analytical method using the approach of Analytical Quality by Design (AQbD). This approach is used when there is a need to reach a full knowledge of the method and, at the same time, to control the possible factors that can influence the results during routine work. The analytical method chosen was the Next Generation Impactor (NGI) that has the capacity of evaluating the aerodynamic particle size distribution (aPSD) of an inhalation product. This method was chosen in order to improve its robustness and, at the same time, to create a workflow representative of the essential steps on its development.The experimental part began after the accomplishment of a risk analysis of the general NGI method using a tool called “Failure Mode and Effect Analysis”. With all the possible critical variables identified, an NGI method was developed and the optimal settings for each variable were defined. The variables tested were: number of capsules discharged per test, time and type of agitation of each component, influence of coating and presence of interstage losses. Among all the tests, it was verified that the mass balance was always within the range recommended by Food and Drug Administration except on the test performed without coating. The variables that had a considerable influence on the results obtained were related with the application of the coating solution and with the time of agitation used during recovery step. Additional tests were performed to evaluate the stability of standards and sample solutions.At the end of the work, it was concluded that the AQbD approach was very useful on the NGI development since it allowed to evaluate which are the method variables with higher impact on the results obtained.