Established biopharmaceuticals such as antibodies and derivatives thereof are relatively large. In cancer therapy, this creates a steep drug concentration gradient within tumors, leaving cells far from blood vessels effectively untreated. Continuous pseudo treatments should foster the development of drug resistance and might lead to eventual disease relapse. Drug concentration gradients can be operationalized as tissue penetration efficiencies, which are functions of molecular size. However, small particles are also subject to potent renal clearance, collapsing the therapeutic window beyond clinical applications. In this master’s thesis, spatial bispecificity was engineered into a single albumin binding domain (ABD). Resulting ABD derived affinity proteins (ADAPTs) are saved from urinary excretion by the grace of HSA, but in the more static microenvironment of tumors, following HSA dissociation, they are capable of tissue penetration efficiencies bestowed only upon smaller particles. To this end, phage display was used to raise ADAPTs against the cancer associated proteins human epidermal growth factor receptor 2 (HER2) and carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6), but also the inflammation marker C-reactive protein (CRP). Via Sanger sequencing, 9 variants were picked for protein production and characterization, among which two spatially bispecific binders were found. ADAPTs were also evaluated for aggregation tendencies, structural conformity to library design, and thermal stability. One ADAPT, binding HER2, passed all tests of initial characterizations. Deep sequencing was used to analyze selection output, from which many more binders should be screened in future experiments. / Etablerade bioläkemedel liksom antikroppar och deras derivat är relativt stora protein. Som cancerterapeutiska skapar de således branta koncentrationsgradienter utgående från tumörpenetrerande blodkärl. Detta riskerar att lämna vissa cancerceller utanför det terapeutiska fönstret. Det svaga selektionstryck som således verkar i tumörperiferin fostrar cancerceller till att utveckla resistens mot detsamma. Koncentrationsgradienten beror på proteinets vävnadspenetrarande förmåga, vilken är en funktion av proteinets storlek. Mindre proteiner borde därmed lättare ackumuleras i hela tumören och förebygga resistensutveckling. Problemet med små proteiner är deras mycket korta halveringstid i serum, en följd av relativt obehindrad filtrering ut i urinen via njurarna. I det här examensarbetet utvecklades rumsbispecifika bindare av cancerassocierade protein med hjälp av fagdisplayselektioner från ett proteinbibliotek baserat på en enda albuminbindande domän (ABD). Resulterande ABD deriverade affinitetsprotein (ADAPT) undkommer ovan nämnda filtrering tack vare sin naturligt starka interaktion med humant serumalbumin (HSA). I den mer långsamt flödande tumörmikromiljön tillåts ADAPTerna efter albumindissociation sedan utöva en bland bioläkemedel överlägsen vävnadspenetration. Tre parallella selektionsspår utfördes mot de cancerassocierade målproteinerna human epidermal growth factor receptor 2 (HER2) och carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6) samt den utsöndrade inflammationsmarkören C-reaktivt protein (CRP). Via Sangersekvensering kunde flera kandidater identifieras. Bland 6 karakteriserade ADAPTer uppvisade samtliga hög HSA-affinitet, tre konstaterades interagera specifikt med sitt målprotein, och två verkade binda även rumsbispecifikt. ADAPTer utvärderades även för sin benägenhet att bilda aggregat, strukturell överensstämmelse med experimentell design, och värmestabilitet. Endast en bindare, mot HER2, klarade sig genom alla prövningar som proteinkarakteriseringen innebar utan underkänt. Även en högparallel sekvensering utav selektionsresultat utfördes, men utanför de tidsramar som tillät ytterligare karakterisering.

No description available.

Cancer is still one of the most common causes of death world-wide and in parallel there is a need to update the repertoire of therapies that withstand resistance of recurrent cancers. Since the introduction of antibody therapies as anti-cancer pharmaceuticals, recognized as immunotherapy in health care, it has been an increasing field in cancer therapy, as a more targeted treatment compared to chemotherapy. Despite the great success, immunotherapy rely on parenteral administration, partly due to poor tissue penetration. If the treatment is administered intravenously, specialized personnel is required, in addition to that it can be inconvenient for the patient. Also, pharmaceuticals based on antibodies often require costly production steps which yields a high-priced treatment. To approach this problem, researchers have developed small affinity domains with the aim to increase tissue penetration while keeping a high specificity to its target. Albumin Binding Domain Derived Affinity Protein (ADAPT) is an example of a small affinity domain of only 7 kDa, which is based on albumin binding domain (ABD) from the streptococcal protein G. Recently, it was shown that the ADAPTs can be further engineered to bind albumin and another relevant target protein of interest simultaneously, which suggests a tolerable half-life in patient serum, alternative administration routes and lower production costs compared to antibody treatments. Furthermore, less side effects are expected due to higher specificity compared to chemotherapy. This work presents the characterization of novel ADAPT proteins that the target the cancer relatedproteins C-C motif ligand 7 (CCL7), vascular endothelial growth factor A (VEGF-A) and carcinoembryonic antigen related cell adhesion molecule 5 (CEACAM5). The new constructs were produced recombinantly in Escherichia coli (E. coli) and purified using affinity chromatography. Moreover, the results demonstrate bispecific binding with high affinity towards serum albumin and CCL7 and CEACAM5 respectively, while the ADAPT variants targeting VEGF-A remain to be further developed. Lastly, the importance of different amino acids for structural and binding properties of one CEACAM5 binder are stated. It reveals that the target binding relies on hydrophobic interactions which also can be connected to its poor structural attributes. Accordingly, this project adds new insights about the ADAPTs which can be useful in research towards future clinical applications aimed to improve cancer treatments.

Detta är ett arbete som analyserar risk- och sårbarhetsanalyser från fyra olika län i Sverige utifrån Béné et al. (2012) teori om resiliens och förmågorna absorbera, anpassa och transformera. Teorin beskriver även begreppet sårbarhet i relation till resiliens och inkluderas därför i den tematiska analysen av risk- och sårbarhetsanalyserna. Arbetet berör det omdiskuterade begreppet resiliens, dess innebörd och applicering inom risk- och krishantering. Undersökningen visar hur olika risk- och sårbarhetsanalyser kan beskriva dessa förmågor och att utformningen av risk- och sårbarhetsanalyserna påverkar hur synliga dessa förmågor är i beskrivningen. Jämförelsen av länen visar att det finns olika förmågor utifrån beskrivningarna som delvis beror på utformningen av risk- och sårbarhetsanalyserna och vilka förutsättningar och erfarenheter som länen har. Länen skilde sig även åt i beskrivningarna av sårbarhet och hur medborgarna beaktades i relation till risker och hot inom länet. Slutsatser som kan dras utifrån arbetet är bland annat att förmågorna absorbera, anpassa och transformera till viss mån beskrivs i risk- och sårbarhetsanalyserna och att principerna i teorin och ramverket som Béné et al. (2012) beskriver också återfinns i olika delar av risk- och sårbarhetsanalyserna. / This is an essay that analyses risk and vulnerability assessments from four different counties in Sweden based on Béné et al. (2012) theory of resilience and the abilities to absorb, adapt and transform. The theory also describes the concept vulnerability in relation to resilience and is included in the thematic analysis of the risk and vulnerability assessments. The work concerns the disputed concept of resilience, its meaning and application in risk and crisis management. The study shows how different risk and vulnerability assessments can describe these abilities and that the design of risk and vulnerability assessments affects how visible these abilities are in the description. The comparison of the counties shows that there are different abilities based on the descriptions, partly due to the design of risk and vulnerability assessment, the conditions and experiences that the counties have. The counties also distinguished themself in the descriptions of vulnerability and how citizens were taken into account in relation to risks and threats within the county. Conclusions that could be drawn from this work is for instance that the abilities to absorb, adapt and transform can be found in the risk and vulnerability assessment and that the principles in the theory and framework that Béné et al. (2012) describes also can be found in various parts of the risk and vulnerability assessments.

As space exploration vehicles travel deeper into space, their distance to earth increases.The increased communication delays and ground personnel costs motivatea migration of the vehicle health management into space. A way to achieve thisis to use a diagnosis system. A diagnosis system uses sensor readings to automaticallydetect faults and possibly locate the cause of it. The diagnosis system usedin this thesis is a model-based reasoning tool called RODON developed by UptimeSolutions AB. RODON uses information of both nominal and faulty behavior ofthe target system mathematically formulated in a model.The advanced diagnostics and prognostics testbed (ADAPT) developed at theNASA Ames Research Center provides a stepping stone between pure researchand deployment of diagnosis and prognosis systems in aerospace systems. Thehardware of the testbed is an electrical power system (EPS) that represents theEPS of a space exploration vehicle. ADAPT consists of a controlled and monitoredenvironment where faults can be injected into a system in a controlled manner andthe performance of the diagnosis system carefully monitored. The main goal of thethesis project was to build a model of the ADAPT EPS that was used to diagnosethe testbed and to generate decision trees (or trouble-shooting trees).The results from the diagnostic analysis were good and all injected faults thataffected the actual function of the EPS were detected. All sensor faults weredetected except faults in temperature sensors. A less detailed model would haveisolated the correct faulty component(s) in the experiments. However, the goal wasto create a detailed model that can detect more than the faults currently injectedinto ADAPT. The created model is stationary but a dynamic model would havebeen able to detect faults in temperature sensors.Based on the presented results, RODON is very well suited for stationary analysisof large systems with a mixture of continuous and discrete signals. It is possibleto get very good results using RODON but in turn it requires an equally goodmodel. A full analysis of the dynamic capabilities of RODON was never conductedin the thesis which is why no conclusions can be drawn for that case.

During the last couple of years there has been a major digitalization of the financial sectorwhich FinTech have largely contributed to. FinTech does not solely contribute withpossibilities, but it also inflicts risks within the financial system when adding challengesrelated to IT-security. These risks pose new operational risks within the financial sector.Therefore, the purpose of this study is to examine as well as analyse different variables thatexplain the perceived trust and risks when using these digital financial services. This isbecause customers play a vital role in keeping their services as well as maintaining financialstability within the FinTech corporations. To examine the subject in this study a deductiveapproach is applied which focuses on previous empirical research and models likeTechnology Acceptance Model (TAM), Elaboration Likelihood Model (ELM) as well asBehavioral finance. These models aim to explain how customers accept new digitalphenomenon. Furthermore, the study performs a quantitative research method through a webbased survey questionnaire. The study examines the independent variables usage, perceivedability to adapt and perceived risk in relation to the dependent factor perceived trust. Thestudy’s sample consists of 185 survey responses which were analyzed through descriptive andinferential statistics. The hypotheses were tested using a multiple regression analysis wherethe independent variables and control variables were analyzed against the dependent variable.The empirical findings showed that customers' perceived ability to adapt to FinTech servicesdid not have a statistically significant effect on perceived trust. Furthermore, perceived riskand usage had a statistically proven influence on perceived trust. The control variables for thisstudy were age and gender where neither had a statistical association to the dependentvariable. Lastly, the empirical findings of the study can be useful for FinTech companiessince it proves that risks need to be minimized to increase perceived trust / Det har skett en stor digitalisering av den finansiella sektorn under de senaste åren därFinTech och dess tjänster har haft en betydande roll. Dock är inte de innovativa digitalatjänsterna som FinTech bidrar till enbart möjligheter, utan det utsätter även den finansiellasektorn för risker. På grund av digitaliseringen medför dessa risker nya operationella riskerinom den finansiella sektorn. Studiens syfte är därav att undersöka och analysera de variablersom förklarar det upplevda förtroendet och riskerna som finns vid användningen av digitalafinansiella tjänster. Studiens fokus ligger på kunderna då de har en nyckelroll i bevarandet avdessa tjänster. Studien har sin grund i ett deduktivt tillvägagångssätt med fokus på tidigareempirisk forskning på området, men även olika modeller som Technology Acceptance Model(TAM), Elaboration Likelihood Model (ELM) samt Behavioral finance som syftar till attförklara hur kunden accepterar nya digitala fenomen. För att undersöka detta tillämpas enkvantitativ metod genom användningen av en webbaserad surveyundersökning. Urvalet bestårav 185 svar som därefter analyserades genom deskriptiv statistik och inferentiell statistik.Detta skedde i form av en multipel regressionsanalys som analyserade studiens tre hypoteserdär studiens tre oberoende variabler och kontrollvariabler analyserades emot studiensberoende variabel. Resultaten visade på att risk och användning hade ett statistiskt sambandmed det upplevda förtroendet medan den egna upplevda förmågan inte hade det. Studiensresultat kan vara användbara för FinTech-företag då resultaten visar att risker måste minskasför att kundförtroendet ska öka.

Marknaden för proteinbaserade läkemedel, de så kallade biologiska läkemedlen, är idag en industri som omsätter miljarder. Ett vanligt sätt att utveckla dessa läkemedel på är med hjälp av monoklonala antikroppar då de kan binda till sitt mål med hög specificitet. Däremot begränsas denna teknik av en lång och dyr produktion som dessutom kräver däggdjursbaserade uttrycksystem. En alternativ teknik till de monoklonala antikropparna är att använda små proteiner som enkelt kan produceras i bakterier till en låg kostnad. Dock begränsas denna metod av de små proteinernas korta cirkuleringstid i blodet. I ett tidigare projekt, har ett litet protein vid namnet ABDderived affinity ProTein (ADAPT) på cirka 7 kDa, utvecklats för att kunna binda till både humant serumalbumin (HSA) för att förlänga cirkulationstiden i blodet och Interleukin 17c (IL17c) som är ett pro-inflammatorisk cytokin. Studien visade dock att ADAPT proteinet inte samtidigt kunde binda till de båda molekylerna tillräckligt effektivt. Syftet med denna uppsats är därför att undersöka om det nämnda proteinet kan optimeras genom så kallad multimering och/eller manipulering av bindningssätet för HSA i syfte att åstadkomma en effektiv och mer långvarig cirkulationstid i blodet samtidigt som det binder sig till sitt mål, IL17c. Tio nya versioner av ADAPT proteinet har utvecklats genom att klona och transformera proteiner till en högt producerande Escherichia coli (E. coli) stam. Proteinerna har sedan producerats och renats fram. Det kunde observeras att proteinerna hade den önskade renheten för att kunna karaktäriseras. Vidare var det möjligt att se att proteinerna hade sin önskade molekylvikt och erhöll sin förväntade struktur som en alfahelix. Proteinernas smältpunkter hade förbättrats eller var liknande jämfört med det ursprungliga proteinet. Dessutom kunde alla proteiner återgå till sin ursprungliga struktur efter upphettning. Utvärderingen av proteinernas bindningskapacitet, med original proteinet som referens, visade på en ökad affinitet till sitt mål, IL17c, för två dimerer och trimeren samt en jämförbar affinitet för två av monomererna med ett manipulerat bindingssäte till HSA. Interaktion till HSA var jämförbar med den ursprungliga ADAPT molekylen för alla nya varianter förutom monomererna med ett manipulerat bindingssäte och dimeren med två manipulerat bindingssäten till HSA. Evaluering av de nya proteinernas kapacitet att binda samtidigt till HSA och IL17c visade att det var gynnsamt med en dimereiserad molekyl då det skapade en distans mellan molekylerna och dess bindningssäten. Vidare kunde det också visas att ordningen som molekylerna interagerade med varandra påverkade proteinernas simultana bindning. / The market for protein-based drugs, or the so-called biopharmaceuticals, is a multibillion-dollar industry today. In the development of protein-based drugs it is common to use monoclonal antibodies (mAbs) due to their ability to bind to its target with high specificity. However, therapeutical development of mAbs is limited by its long and expensive production in mammalian expression system. An alternative to mAbs are the so-called alternative scaffolds which are small proteins that can be produced in bacteria at lower costs. Although a drawback with the latter proteins is their short serum half-life. A small scaffold protein, ABD-Derived Affinity ProTein (ADAPT) of approximate 7 kDa was earlier engineered to obtain bispecific affinity, to Human Serum Albumin (HSA), to extend its half-life, as well as to the pro-inflammatory cytokine, Interleukin 17c (IL17c). Unfortunately, it was shown that the simultaneous binding was not efficient enough for its desired purpose. The aim with this project was therefore to investigate if the previous mentioned binder could be optimized by multimerization and/or manipulation of the HSA binding site for an efficient half-life extension. By generating ten new designs of the ADAPT variants, it was observed that the new variants had stable alpha helical structures and an improved or similar melting temperature as the original variant. The evaluation of the target binding displayed an improved affinity to the target, IL17c, for two of the dimeric versions as well as for the trimer and a comparable affinity for two of the monomers with a manipulated HAS binding site. The interaction to HSA was comparable to the original ADAPT for all binders except from the monomers with impaired HSA binding and the dimer with two impaired HSA binding sites. The evaluation of the simultaneous binding showed that it was favored by dimerization when a distance between the two molecule and their binding surfaces was added. Moreover, it could also be seen that the order of binding events had an impact on the simultaneous binding.

Under senare år har målinriktad terapi varit ett växande område inom cancerterapi som en mer målinriktad behandling än kemoterapi. Dessa behandlingar baseras främst på antikroppsbaserade läkemedel som är ganska stora och komplexa, vilket ökar den totala kostnaden för behandlingen. Därför måste man hitta en alternativ metod för både upptäckt och behandling för att hjälpa patienterna. Små affinitetsdomäner har skapats med målet att förbättra vävnadspenetrationen och samtidigt upprätthålla en hög grad av målspecificitet, vilket leder till färre biverkningar. Ett av exemplen på detta är Albumin Binding Domain-Derived Affinity Protein (ADAPT). Det har baserats på en av de albuminbindande domänerna (ABD) i streptokockproteinet G, med en storlek på 6,5 kDa. Nyligen modifierades ADAPT ytterligare för att samtidigt binda albumin och ett annat relevant målprotein av intresse, vilket tyder på en längre halveringstid i patientserum och möjliggör utveckling av nyare och terapeutiska läkemedel. I detta projekt presenteras den fjärde generationen av ADAPT-biblioteket som utformats för att ha förbättrad stabilitet. Selektioner med fagdisplay utfördes mot tre målproteiner: carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5), en biomarkör för kolorektalcancer, epithelial cell adhesion molecule (EpCAM), en markör för flera gastrointestinala karcinom och trophoblast cell-surface antigen 2 (Trop2) som är överuttryckt i trippel-negativ bröstcancer. Resultaten visar bindning till CEACAM5, EpCAM och Trop2, vilket har visats med monoklonal fag-ELISA. Bindningsaffiniteten, sekundärstrukturen hos de utvalda bindarna och den bispecifika bindningen till serumalbumin återstår att utvärdera ytterligare. Projektet visar således att de ADAPTs som valts ut mot målen CEACAM5, EpCAM och Trop2 har en enorm potential för framtida kliniska tillämpningar som syftar till utveckling av diagnostik och terapi för dessa cancerbiomarkörer. / In recent years, targeted therapy has been a growing field of cancer therapy as a more targeted treatment than chemotherapy. These treatments are primarily based on antibody-based pharmaceuticals which are quite large and complex, increasing the overall cost of the treatment. Thus, an alternative method of both detection and treatment needs to be found to aid patients. Small affinity domains have been created with the goal of enhancing tissue penetration while maintaining a high level of target specificity, leading to fewer side effects. One of the examples for these is the Albumin Binding Domain-Derived Affinity Protein (ADAPT). It has been based on one of the albumin binding domains (ABD) of the streptococcal protein G, with a size of 6.5 kDa. Recently, the ADAPTs were further modified to simultaneously bind albumin and another pertinent target protein of interest, suggesting a longer half-life in patient serum, and enabling the development of newer therapeutics. This project presents the 4th generation of the ADAPT library designed to have improved stability. Phage display selections were performed against three target proteins: carcinoembryonic antigen- related cell adhesion molecule 5 (CEACAM5), a biomarker for colorectal cancer, epithelial cell adhesion molecule (EpCAM), a marker for several gastrointestinal carcinomas and trophoblast cell-surface antigen 2 (Trop2) which is overexpressed in triple-negative breast cancer. The results demonstrate binding towards CEACAM5, EpCAM and Trop2, which has been shown by monoclonal phage ELISA. The binding affinity, secondary structure of the selected binders and bispecific binding towards serum albumin remain to be further assessed. The project thus reveals that the ADAPTs selected against the targets CEACAM5, EpCAM and Trop2 present a massive potential for future clinical applications aimed towards development of diagnostics and therapeutics for these cancer biomarkers.

Submitted by SBI Biblioteca Digital (sbi.bibliotecadigital@puc-campinas.edu.br) on 2018-08-15T11:11:40Z No. of bitstreams: 1 Fabio Nunes Angelo.pdf: 3745845 bytes, checksum: c448700fa075a4f93f9dc10fb0233246 (MD5) / Made available in DSpace on 2018-08-15T11:11:40Z (GMT). No. of bitstreams: 1 Fabio Nunes Angelo.pdf: 3745845 bytes, checksum: c448700fa075a4f93f9dc10fb0233246 (MD5) Previous issue date: 2007-05-31 / Centro de Pesquisa e Desenvolvimento em Telecomunica??es (CPqD) / The subject of this thesis is an analysis on the effect of information granularity in adaptive hypermedia systems upon the response time in the interaction with the user. First the concept of adaptive hypermedia is brought into consideration, followed by the concept of information granularity itself. These concepts are then related to adaptive hypermedia system in order to measure the possible performance problems that can rise when information granularity is modified. Next, a methodology for performance assessment of computer systems is presented. This methodology is adapted and used to study the impact of refining information granularity upon the performance of an adaptive hypermedia system. Information is presented on the mean response time data that was collected and modeled in the form of mathematical equations, where the amount of adaptive application objects are related to average response time. An specific estimation tool was developed using these equations. The issue on performance versus optimal size for information granularity in adaptive hypermedia systems is discussed and.the conclusion suggests new directions for future work. / O assunto desta disserta??o ? uma an?lise sobre o impacto da varia??o da granularidade da informa??o em sistemas hiperm?dia adapt?veis em cima do tempo de resposta na intera??o com o usu?rio. Discute-se primeiramente o conceito de hiperm?dia adapt?vel, seguido pelo conceito de granularidade da informa??o. Estes conceitos s?o, ent?o, relacionados ao problema de desempenho de uma aplica??o hiperm?dia adapt?vel, quando ocorre mudan?a na granularidade da informa??o. Em seguida, apresenta-se uma metodologia para avalia??o de desempenho de sistemas computacionais. Atrav?s de algumas adapta??es, a mesma ? aplicada para estudar o impacto do refinamento da granularidade da informa??o no desempenho de um determinado sistema hiperm?dia adapt?vel. Apresentam-se os dados sobre tempo m?dio de resposta e modelam-se os mesmos em equa??es, nas quais relaciona-se a quantidade de objetos da aplica??o adapt?vel com o tempo m?dio de resposta. Uma ferramenta de estimativa desenvolvida a partir dessas equa??es ? apresentada. Discute-se at? que ponto a informa??o deve ser refinada nos sistemas hiperm?dia adapt?veis sem que seu desempenho seja afetado e conclui-se sugerindo uma linha de trabalhos futuros.

Le développement d’un médicament est non seulement complexe mais les retours sur investissment ne sont pas toujours ceux voulus ou anticipés. Plusieurs médicaments échouent encore en Phase III même avec les progrès technologiques réalisés au niveau de plusieurs aspects du développement du médicament. Ceci se traduit en un nombre décroissant de médicaments qui sont commercialisés. Il faut donc améliorer le processus traditionnel de développement des médicaments afin de faciliter la disponibilité de nouveaux produits aux patients qui en ont besoin. Le but de cette recherche était d’explorer et de proposer des changements au processus de développement du médicament en utilisant les principes de la modélisation avancée et des simulations d’essais cliniques. Dans le premier volet de cette recherche, de nouveaux algorithmes disponibles dans le logiciel ADAPT 5® ont été comparés avec d’autres algorithmes déjà disponibles afin de déterminer leurs avantages et leurs faiblesses. Les deux nouveaux algorithmes vérifiés sont l’itératif à deux étapes (ITS) et le maximum de vraisemblance avec maximisation de l’espérance (MLEM). Les résultats de nos recherche ont démontré que MLEM était supérieur à ITS. La méthode MLEM était comparable à l’algorithme d’estimation conditionnelle de premier ordre (FOCE) disponible dans le logiciel NONMEM® avec moins de problèmes de rétrécissement pour les estimés de variances. Donc, ces nouveaux algorithmes ont été utilisés pour la recherche présentée dans cette thèse. Durant le processus de développement d’un médicament, afin que les paramètres pharmacocinétiques calculés de façon noncompartimentale soient adéquats, il faut que la demi-vie terminale soit bien établie. Des études pharmacocinétiques bien conçues et bien analysées sont essentielles durant le développement des médicaments surtout pour les soumissions de produits génériques et supergénériques (une formulation dont l'ingrédient actif est le même que celui du médicament de marque, mais dont le profil de libération du médicament est différent de celui-ci) car elles sont souvent les seules études essentielles nécessaires afin de décider si un produit peut être commercialisé ou non. Donc, le deuxième volet de la recherche visait à évaluer si les paramètres calculer d’une demi-vie obtenue à partir d'une durée d'échantillonnage réputée trop courte pour un individu pouvaient avoir une incidence sur les conclusions d’une étude de bioéquivalence et s’ils devaient être soustraits d’analyses statistiques. Les résultats ont démontré que les paramètres calculer d’une demi-vie obtenue à partir d'une durée d'échantillonnage réputée trop courte influençaient de façon négative les résultats si ceux-ci étaient maintenus dans l’analyse de variance. Donc, le paramètre de surface sous la courbe à l’infini pour ces sujets devrait être enlevé de l’analyse statistique et des directives à cet effet sont nécessaires a priori. Les études finales de pharmacocinétique nécessaires dans le cadre du développement d’un médicament devraient donc suivre cette recommandation afin que les bonnes décisions soient prises sur un produit. Ces informations ont été utilisées dans le cadre des simulations d’essais cliniques qui ont été réalisées durant la recherche présentée dans cette thèse afin de s’assurer d’obtenir les conclusions les plus probables. Dans le dernier volet de cette thèse, des simulations d’essais cliniques ont amélioré le processus du développement clinique d’un médicament. Les résultats d’une étude clinique pilote pour un supergénérique en voie de développement semblaient très encourageants. Cependant, certaines questions ont été soulevées par rapport aux résultats et il fallait déterminer si le produit test et référence seraient équivalents lors des études finales entreprises à jeun et en mangeant, et ce, après une dose unique et des doses répétées. Des simulations d’essais cliniques ont été entreprises pour résoudre certaines questions soulevées par l’étude pilote et ces simulations suggéraient que la nouvelle formulation ne rencontrerait pas les critères d’équivalence lors des études finales. Ces simulations ont aussi aidé à déterminer quelles modifications à la nouvelle formulation étaient nécessaires afin d’améliorer les chances de rencontrer les critères d’équivalence. Cette recherche a apporté des solutions afin d’améliorer différents aspects du processus du développement d’un médicament. Particulièrement, les simulations d’essais cliniques ont réduit le nombre d’études nécessaires pour le développement du supergénérique, le nombre de sujets exposés inutilement au médicament, et les coûts de développement. Enfin, elles nous ont permis d’établir de nouveaux critères d’exclusion pour des analyses statistiques de bioéquivalence. La recherche présentée dans cette thèse est de suggérer des améliorations au processus du développement d’un médicament en évaluant de nouveaux algorithmes pour des analyses compartimentales, en établissant des critères d’exclusion de paramètres pharmacocinétiques (PK) pour certaines analyses et en démontrant comment les simulations d’essais cliniques sont utiles. / Drug development is complex with results often differing from those anticipated or sought after. Despite technological advances in the many fields which are a part of drug development, there are still many drugs that fail in the late stages of clinical development. Indeed, the success rate of drugs reaching commercialization is declining. Improvements to the conventional drug process are therefore required in order to facilitate development and allow new medications to be provided more rapidly to patients who require them. The aim of this Ph.D. project was to explore and propose ways to improve this inefficient drug development process with the use of advanced modeling and clinical trial simulations. For the first part of this research, new algorithms available in ADAPT 5® were tested against other available algorithms in order to determine their potential strengths and weaknesses. The two new algorithms tested were the iterative two-stage and the maximum likelihood expectation maximization (MLEM) methods. Our results demonstrated that the MLEM algorithm was consistently better than the iterative two-stage algorithm. It was also comparable with the first order conditional estimate method available in NONMEM®, with significantly fewer shrinkage issues in the estimation of the variances. Therefore, these new tools were used for the clinical trial simulations performed during the course of this Ph.D. research. In order to calculate appropriate noncompartmental pharmacokinetic parameter estimates during the drug development process, it is essential that the terminal elimination half-life be well characterized. Properly conducted and analyzed pharmacokinetic studies are essential to any drug development plan, and even more so for generic and supergeneric (a formulation similar to the reference product, containing the same active ingredient; however differing from the original reference product it its delivery process) submission where they often are the only pivotal studies that need to be done to decide if a drug product is good or not. Thus, the purpose of the second part of the research was to determine if the pharmacokinetic (PK) parameters obtained from a subject whose half-life is calculated from a sampling scheme duration that is considered too short could bias the bioequivalence conclusions of a study and if these parameters should be removed from statistical analyses. Results demonstrated that subjects with a long half-life relative to the duration of the sampling scheme negatively influenced results when these were maintained in the analysis of variance. Therefore, these subjects should be removed from the analyses and guidelines to this effect are required a priori. Pivotal pharmacokinetic studies done within the drug development process should therefore follow this recommendation to make sure that the right decision be taken on a drug product formulation. This information was utilized with the clinical trial simulations that were subsequently performed in this research in order to ensure the most accurate conclusions. Finally, clinical trial simulations were used to improve the development process of a nonsteroidal anti-inflammatory drug. A supergeneric was being developed and results from a pilot study were promising. However, some results from the pilot study required closer attention to determine if the test and reference compounds were indeed equivalent and if the test compound would meet the equivalence criteria of the different required pivotal studies. Clinical trial simulations were therefore undertaken to address the multiple questions left unanswered by the pilot study and these suggested that the test compound would probably not meet the equivalence criteria. In addition, these results helped determine what modifications to the test drug would be required to meet the equivalence criteria. This research brought forward solutions to improve different aspects of the drug development process. Notably, clinical trial simulations reduced the number of studies that would have been done for a supergeneric, decreased the number of subjects unnecessarily exposed to a drug, lowered costs and helped established new criteria for the exclusion of subjects from analyses. Research conducted during this Ph.D. provided concrete ways to improve the drug development process by evaluating some newly available tools for compartmental analyses, setting standards stipulating which estimated PK parameters should be excluded from certain PK analyses and illustrating how clinical trial simulations are useful to throughout the process.