Thesis topic concerns the issue of information security in corporate environments. Literature search includes information obtained by studying articles and literature in the field of information security. Resources were selected with a focus on the security risks, security technologies and legislative regulation. Attention is focused on technology that supports monitoring of communication flows in the data network. Overview of traffic operating a data network provides important information for the prevention or investigation of security incidents. Monitoring also serves as a source of information for the planning of the network infrastructure. It can detect faults or insufficient transmission capacity. The practical part is dedicated to implementation of the monitoring system in the real corporate networks. Part of the experience is the analysis of the network structure and choice of appropriate tools for actual implementation. When selecting tools, you can use the scoring method of multicriterial analysis options. The integration of the monitoring system is also the configuration of active network elements. Subsequent analysis of network traffic provides information about the most active users, most used applications or on the sources and targets of data transmitted. It provides a source of valuable information that can be used in case of failure on the network or security incident. The conclusion is a summary of the results and workflow.

Avec les progrès technologiques réalisés au cours de ces dernières décennies, la complexité et les vitesses de fonctionnement des circuits intégrés ont beaucoup été augmentées. Bien que ces évolutions aient permis de diminuer les dimensions et les tensions d’alimentations des circuits, la compatibilité électromagnétique (CEM) des composants a fortement été dégradée. Identifiée comme étant un verrou technologique, la CEM est aujourd’hui l’une des principales causes de « re-design » des circuits car les problématiques liées aux mécanismes de génération et de couplage du bruit ne sont pas suffisamment étudiées lors de leur conception.Ce manuscrit présente donc une méthodologie visant à étudier la propagation du bruit à travers les circuits intégrés par mesures et par simulations. Afin d’améliorer nos connaissances sur la propagation d’interférences électromagnétiques (IEM) et les mécanismes de couplage à travers les circuits, nous avons conçu un véhicule de test développé dans la technologie SMOS8MV® 0,25 µm de Freescale Semiconductor. Dans ce circuit, plusieurs fonctions élémentaires telles qu’un bus d’E/S et des blocs numériques ont été implémentées. Des capteurs de tensions asynchrones ont également été intégrés sur différentes alimentations de la puce pour analyser la propagation des perturbations injectées sur les broches du composant (injection DPI) et sur les conducteurs permettant d’alimenter ce dernier (injection BCI). En outre, nous proposons différents outils pour faciliter la modélisation et les simulations d’immunité des circuits intégrés (extraction des modèles de PCB, approches de modélisation des systèmes d’injection, méthode innovante permettant de prédire et de corréler les niveaux de tension/ de puissance injectés lors de mesures d’immunité conduite, flot de modélisation). Chaque outil et méthode de modélisation proposés sont évalués sur différents cas test. Enfin, pour évaluer notre démarche de modélisation, nous l’appliquons sur un bloc numérique de notre véhicule de test et comparons les résultats de simulations aux différentes mesures internes et externes réalisées sur le circuit / With technological advances in recent decades, the complexity and operating speeds of integrated circuits have greatly increased. While these developments have reduced dimensions and supply voltages of circuits, electromagnetic compatibility (EMC) of components has been highly degraded. Identified as a technological lock, EMC is now one of the main causes of circuits re-designs because issues related to generating and coupling noise mechanisms are not sufficiently studied during their design. This manuscript introduces a methodology to study propagation of electromagnetic disturbances through integrated circuits by measurements and simulations. To improve our knowledge about propagation of electromagnetic interferences (EMI) and coupling mechanisms through integrated circuits, we designed a test vehicle developed in the SMOS8MV® 0.25µm technology from Freescale Semiconductor. In this circuit, several basic functions such as I/O bus and digital blocks have been implemented. Asynchronous on-chip voltage sensors have also been integrated on different supplies of the chip to analyze propagation of disturbances injected on supply pins and wires of the component (DPI and BCI injection). In addition, we propose various tools to facilitate modeling and simulations of Integrated Circuit’s immunity (PCB model extraction, injection systems modeling approaches, innovative method to predict and correlate levels of voltage / power injected during conducted immunity measurements, modeling flow). Each tool and modeling method proposed is evaluated on different test cases. To assess our modeling approach, we finally apply it on a digital block of our test vehicle and compare simulation results to various internal and external measurements performed on the circuit

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.

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.

Certaines applications issues des biotechnologies sont actuellement controversées d’un point de vue sociotechnique. A l’analyse, il s’avère que les protagonistes de ces controverses utilisent des arguments évolutionnistes de façon récurrente à propos de questions très diverses et parfois de façon apparemment contradictoire. Afin de comprendre les raisons d’un tel usage, d’une part, nous avons procédé dans cette thèse à une analyse de discours des controverses relatives au diagnostic préimplantatoire (DPI) et aux OGM agricoles à partir d’essais et d’articles issus de revues spécialisées ou de vulgarisation. D’autre part, nous avons effectué une étude de l’histoire de l’évolutionnisme scientifique et non scientifique. Nous avons ainsi pu établir que se rejoue à travers ces controverses sociotechniques une partie des controverses structurantes de la pensée évolutionniste et que l’évolutionnisme constitue un cadre de représentations prégnant permettant de débattre collectivement des nouvelles techniques du vivant. / Some biotechnologies are currently socially debated. One may notice that the protagonists of the controversies make regular use of evolutionist warrants concerning very diverse subjects and sometimes in an apparently contradictory way. In order to understand this fact, on the one hand, we analysed controversies relating to preimplantation genetic diagnosis (PGD) and GMOs in agriculture on the basis of some essays, review papers and newspaper articles. On the other hand, we studied the history of scientific and non-scientific evolutionist theories. Thus, we established that some essential controversies of evolutionism were repeated into the social controversies about PGD and GMOs and that evolutionism represents a meaningful framework in which new biotechnologies can be collectively debated.

The aim of this thesis is to analyze various kinds of co-simulation techniques and to design integration these techniques into Lissom simulation platform. The first section of this thesis shows capabilities of external interfaces of simulation platforms for HDL languages VHDL, Verilog and SystemVerilog and of tool Matlab. The second section deals with design of synchronization mechanism among Lissom simulation platform and others simulation platforms using mentioned external interfaces. In the last part the testing of implemented solutions and evaluation of results is described.

No description available.

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.

Le calcul réparti est de plus en plus utilisé bien qu'il reste très mal maîtrisé. Cette thèse porte sur le Dpi-calcul, une extension simple du pi-calcul dans laquelle tous les processus sont placés dans des localités afin de décrire leur répartition. Dans ce calcul, les processus peuvent communiquer localement et migrer entre localités. À côté des canaux de communication et des localités, on identifie une nouvelle famille d'identifiants, les passeports, permettant un contrôle fin des migrations de processus : un processus doit disposer d'un passeport adéquat pour entrer dans une localité.<br /><br />Afin de structurer le calcul, on met en place un système de types qui associe un type à chaque identifiant pour vérifier qu'un processus n'utilise que les droits qu'il possède. L'ordre de sous-typage sur les types est étendu aux types de passeports suivant les localités d'origine des processus migrant. On démontre que cet ordre admet des bornes inférieures sous certaines conditions. On prouve également que les processus se conformant à cette politique de typage conservent cette propriété au cours de leurs réductions.<br /><br />On étudie aussi l'équivalence observationnelle : quand des processus exhibent-ils des comportements indiscernables pour un observateur ? En présence de passeports, il est indispensable d'imposer à l'observateur d'être loyal, c'est-à-dire d'exiger la possession de passeports pour observer les communications ayant lieu dans les localités correspondantes. Ces contraintes définissent une congruence dite barbue loyale. On développe ensuite un système de transitions étiquetées tel que la bisimilarité loyale engendrée coïncide avec cette congruence barbue loyale.

A number of therapeutic proteins are used for long in clinical practice. These include for example insulin, calcitonine, growth hormone, and parathyroid hormone for the treatment of various systemic disorders, as well as protein antigens in vaccine formulations. Due to the recent developments in biochemical engineering and in the comprehension of the physiopathology of many diseases, peptides and proteins are expected to become a drug class of increasing importance. Recently, novel biological drugs have for example been developed such as monoclonal antibodies, antibody fragments, soluble receptors, and receptor agonists or antagonists. These are mainly used for the treatment of auto-immune and inflammatory diseases (asthma, rheumatoid arthritis) and for the treatment of cancers. However, a major drawback of these biomolecules is the need to use parenteral administration. This is mainly due to the harsh pH conditions that proteins undergo by oral administration, leading to various physico-chemical degradations and loss of biological activity. <p><p>Pulmonary delivery of these proteins could constitute an alternative to parenteral delivery. Due to the very high surface area of the lungs, the low thickness of the alveolar epithelium and the high level of lung vascularisation, pulmonary administration can indeed provide fast systemic absorption of drugs, while avoiding hepatic first pass metabolism. On the other hand, drugs for local treatment can also be administered directly into the lung, which allows delivering high doses while limiting systemic side effects. Nevertheless, administration of drugs to the lungs requires some challenges to be taken up. It is indeed necessary to provide the drug as very small solid or liquid microparticles (1-5 µm) in order to reach the lungs. For solid microparticles, it is also needed to overcome the very high inter-particle interactions by using appropriate formulation strategies and by including deaggregation mechanisms in the inhalation device. Other issues are more specifically related to the pulmonary administration of proteins. These can indeed undergo physico-chemical degradations during processing, administration, and/or storage. Moreover, if systemic action is required, proteins will often need addition of an absorption enhancer to cross the alveolar epithelium because of their large molecular weight and hydrophilicity. <p><p>In this work, we developed formulations for pulmonary delivery of proteins using two model proteins. Insulin (5.8 kDa) was chosen as a model of small protein. It is also an application of systemic pulmonary delivery. On the other hand, an anti-IL13 monoclonal antibody fragment (54 kDa) was used as a model of larger protein. This molecule is currently in development for the treatment of asthma and provided an application for local pulmonary delivery. The formulation strategy was to produce dry powders using a combination of micronisation techniques (high speed and high pressure homogenisations), drying techniques (spray-drying, freeze-drying), and addition of lipid excipients. These lipid excipients were added as a coating around the protein particles and were expected to prevent protein degradations during processing and/or storage, essentially by avoiding contact with water. It could also improve the aerodynamic properties of the powders by modification of the surface properties of the particles and/or limitation of the capillary forces.<p><p>First, we evaluated insulin lipid-coated formulations and formulations without excipients, produced using high pressure homogenisation and spray-drying. In the case of lipid-coated formulations, a physiological lipid composition based on a mixture of cholesterol and phospholipids was used. We were able to obtain good aerodynamic features for the different formulations tested, with fine particle fractions between 46% and 63% versus 11% for raw insulin powder. These are high FPF values in comparison with those obtained for other protein formulations for inhalation currently under development, which often have an in vitro deposition of around 30%. Insulin presented a good stability in the dry state, even when no lipid coating was added.<p>The presence of a lipid coating of up to 30% (w/w) did not significantly improve the aerodynamic behaviour of the powders, but the coated formulations exhibited decreased residual moisture content after 3-month storage, which should be of interest for the long-term stability of the formulations. <p><p>In a second step, two of the developed insulin formulations were evaluated in a clinical study to determine whether the formulations give high deep lung deposition in vivo, and how insulin is absorbed into the systemic blood stream. This pharmaco-scintigraphic trial was performed on twelve type 1 diabetic patients using an uncoated formulation and a formulation coated with 20% (w/w) of lipids. The two formulations showed interesting features, with pharmacokinetic profiles that mimic the natural insulin secretion pattern. Bioavailability was within the ranges of two of the three dry powder insulins that have reached phase III clinical development. However, the formulation with a lipid coating exhibited a lower lung deposition in comparison with the uncoated formulation, which was not expected from the previous in vitro results. Additional in vitro experiments indicated that this lower performance was related to a decrease in the disaggregation efficiency of the powder at a sub-optimal inhalation flow-rate. An extensive training of the patients to the inhalation procedure could therefore improve the lung deposition of the coated formulation.<p><p>Finally, we developed and evaluated dry powder formulations of the anti-IL13 antibody fragment. These were produced using, successively, freeze-drying, high pressure homogenisation (HPH), and spray-drying. The influence of different types and concentrations of stabilising excipients was evaluated for each production step. Due to its more elaborated structure, the antibody fragment was found to be more sensitive than insulin to physico-chemical degradation, particularly during the HPH process, which led to different types of degradation products. These could partly be avoided by adding 50% sucrose during freeze-drying and 10% Na glycocholate or palmitic acid in the liquid phase during HPH (dispersing agents). However, the presence of a small fraction of insoluble aggregates could not be fully avoided. Further spray-drying of the suspensions in the presence of 10% Na glycocholate or palmitic acid led to the formation of a hydrophilic or hydrophobic coating around the particles, respectively. Na glycocholate was found to be particularly effective in protecting the antibody during spray-drying, which was found to be at least partly related to its ability to inhibit sucrose recrystallisation. However, the best formulation still presented a small fraction of insoluble aggregates (6%). The aerodynamic evaluation of the formulations showed FPFs that were compatible with lung deposition, with the formulation containing Na glycocholate presenting the highest FPF (42%). The formulation coated with palmitic acid presented a slightly lower FPF (35%). The aerodynamic properties of this formulation remained unchanged at a sub-optimal inspiratory flow rate, to the contrary of what was observed for the insulin formulation coated with 20% (w/w) cholesterol and phospholipids. Palmitic acid could therefore be of interest as a hydrophobic coating material, and provide long-term stability of protein drugs. <p>The work performed with the insulin and anti-IL13 molecules provided the proof-of-concept that it was possible to obtain dry powder protein formulations with appropriate aerodynamic properties and good overall physico-chemical stability, using simple production techniques and few selected excipients. The formulation strategy presented in this work could therefore be of interest for the future development of inhaled proteins for local or systemic applications. <p> / Doctorat en sciences pharmaceutiques / info:eu-repo/semantics/nonPublished