<p>Antibody-drug conjugates (ADCs) combine the cytotoxicity of traditional chemotherapy with the site-specificity of antibodies by conjugating payloads to antibodies with immunoaffinity. However, the conjugation alters the physicochemical properties of antibodies, increasing the risks of various types of degradation. The effects of common risk factors such as pH, temperature, and light on the stability of ADCs differ from their effects on monoclonal antibodies (mAb) due to these altered physicochemical properties. </p> <p>To date, ADC researchers have developed linkers with improved <i>in vivo</i> stability, and begun to understand the deconjugation mechanisms <i>in vivo</i>. In contrast, the <i>in vitro</i> stability of ADCs has not gained comparable attention. All nine of the U.S. FDA approved ADCs are lyophilized to minimize the potential for degradation. However, there are few studies on the solid-state stability of ADCs. To evaluate lyophilized solids, pharmaceutical development relies heavily on accelerated stability studies, which take months to determine the best formulation. Characterization methods that are often used orthogonally with accelerated studies include Fourier-transform infrared spectroscopy (FT-IR), Raman spectroscopy, near-infrared spectroscopy (NIR), differential scanning calorimetry (DSC), and x-ray powder diffraction (XRPD). Results from these methods are often poorly correlated with stability, however. Thus, stability evaluation of solid-state ADC products, and other recombinant protein drugs, is often a bottleneck in their development.</p> <p>To provide knowledge on how to improve the <i>in vitro</i> stability of lyophilized ADC formulations, the solid-state stability of ADC formulations with varying risk factors was studied in this dissertation project. The first study investigated interactions between an ADC and excipients in terms of solid-state stability enhancement. The second study investigated the process-driven instability of ADCs during lyophilization using various concentrations of ADCs. The first two studies incorporate a new method called solid-state hydrogen/deuterium exchange coupled with mass spectrometry (ssHDX-MS) as an analytical predictor of solid-state stability. The last study investigated the effects of pH on the stability of labile hydrazones, as a model for common linker chemistry used in ADCs. </p>

En el Perú, la minería es una de las actividades importantes en el desarrollo del crecimiento económico del país, sin embargo, el uso indiscriminado de los recursos naturales y el impacto ambiental, debido a la extracción del mineral, obligó a las entidades reguladoras como el ANA, OEFA, OSINERGMIN a estandarizar los procesos de extracción y operación. Esto contempla estabilizar ambientalmente los componentes que se encuentran dentro del proceso de extracción del mineral, además, de estabilizar, física, química e hidrológicamente a aquellos que, una vez agotado el recurso (mineral), se transforman en un pasivo que deben ser incorporados de manera estable al medio ambiente (cierre de pasivo). La presente tesis propone evaluar la condición de estabilidad actual del depósito de desmonte de roca N8-PO. La evaluación se realizará usando programación lineal a través del método Simus que recoge los parámetros más importantes para cada condición de estabilidad actual, analizado en el pasivo, el cual será comparada con los factores de seguridad requeridos por las entidades reguladoras y las buenas prácticas de construcción. Para el análisis de estabilidad física se recogerán los resultados del análisis estático y pseudo estático, para el análisis de estabilidad geoquímica, se recogerán los resultados de los ensayos ABA, finalmente, para el análisis de estabilidad hidrológica, se tendrá en cuenta el estudio hidrológico, además de la existencia y condición de estructuras hidráulicas presentes. Los resultados de la evaluación indicarán la condición de estabilidad del pasivo. Por último, los resultados recomendarán también, posibles alternativas de cierre a usar. / In Peru, mining is one of the important activities in the development of the country's economic growth, however, the indiscriminate use of natural resources and the environmental impact, due to mineral extraction, forced regulatory entities such as ANA, OFEA, OSINERGMIN to standardize the extraction and operation processes. This includes not only environmentally stabilizing the components that are within the mineral extraction process, but also, physically, chemically and hydrologically stabilizing those who, once the (mineral) resource has been depleted, become a responsibility that must Stably incorporate into the environment (passive closure). This thesis proposes to evaluate the current stability condition of the N8-PO rock dump and compare with the results of the evaluation that will be carried out with two additional components, of the same characteristics, located near the component under study. The evaluation will be carried out through a program that uses a numerical method that collects the most important parameters for each stability condition, current, analyzed in the dump, which will be compared with the safety factors required by regulatory entities and Good construction practices. In the analysis of the physical stability the results of the static and pseudostatic analysis will be collected, for the analysis of the geochemical stability, the results of the ABA tests will be collected, finally, for the analysis of the hydrological stability, the studies will be taken Hydrology account, as well as the existence and condition of the hydraulic structures present. The evaluation results will indicate the stability condition of the mining component. Finally, the results will also recommend possible closure alternatives that will be used to achieve the desired stability. / Tesis

No / Sedimentation of non-homogeneous systems is the typical phenomenon indicating the physical instability as a key measure to the quality control of the preparation products. Currently, the determination methods for the sedimentation of non-homogeneous preparations are based on manual measurement or semi-quantitative observation, lacking of either automation or quantitative dynamic analysis. The purpose of this research was to realize automatic and quantitative monitoring of the sedimentation dynamics for non-homogenous systems as suspension, emulsions at laboratory level. Non-contact measurement method has been established to determine the sedimentation behaviors in a standard quartz tube for sedimentation, with internal diameter and height 23 mm and 215 mm, respectively, with controlled temperature and light intensity. As high performance camera has been equipped, the sedimentation images with high spatial and temporal resolution could be acquired, which can continuously capture sedimentation images with the resolution of 2048 x 2048 pixel at a maximum rate of 60 slides/s. All the pictures were processed to extract the luminance matrix top-down along the fixed vertical midline of each picture, which implied sedimentation characteristics of the system at the moment the picture was taken. Combining all the luminance matrixes along vertical middle lines of the pictures, a time-luminance matrix profile was obtained. Digital image processing techniques were used to eliminate interference and establish a three-dimensional surface model of the sedimentation dynamics. Then, the derivative mutation algorithm has been developed for the intelligent identification of sedimentation interface with threshold optimization so as to quantitatively analyze the sedimentation dynamics with visualization. The sedimentation curve and sedimentation dynamic equation of the non-homogeneous system were finally outputted by numerical fitting. The methodology was validated for great significance in determinations of small volume samples, parallel control multiple batches, and long period of time automatic measurement. (C) 2015 Elsevier B.V. All rights reserved.

Le développement d’un médicament pour une cible thérapeutique donnée passe par plusieurs étapes qui se résument en une étape de criblage, une phase préclinique et plusieurs phases cliniques. Ces étapes permettent de sélectionner une substance active et de démontrer son efficacité thérapeutique et sa sécurité toxicologique. Ces deux critères définissent la qualité du médicament qui, une fois démontrée, doit être garantie pendant toute sa durée de validité. La qualité est évaluée au moyen d’études de stabilité qui sont réalisées d’abord sur la matière première de la substance active au cours de la phase de pré-développement du médicament, ensuite sur le produit fini. La stabilité intrinsèque de la substance active concerne à la fois ses propriétés chimiques et ses propriétés physiques qui sont liées à la nature de la substance. L’étude de stabilité repose d’abord sur la caractérisation de ces propriétés, et ensuite sur l’étude de la sensibilité de la substance à l’égard des facteurs environnementaux pouvant modifier les propriétés intrinsèques de la substance. L’approche adoptée dans ce travail repose d’une part sur l’évaluation de la stabilité chimique c’est à dire de la réactivité chimique des substances à usage pharmaceutique au travers des études de pureté chimique et des études de dégradation forcée de ces substances en solution, et d’autre part, sur l’évaluation de la stabilité physique. Dans ce cadre, l’étude du polymorphisme cristallin revêt une grande importance, tout comme l’aptitude à la formation d’hydrates ou de solvates. Cette étude, basée sur la thermodynamique, consiste pour l’essentiel à construire un diagramme de phases pression-température permettant de définir les domaines de stabilité relative des différentes formes cristallines. Cinq substances actives, existant à l’état solide et entrant dans la composition de médicaments administrés par voie orale, ont été étudiées dans le cadre de ce travail. L’analyse chimique du tienoxolol, présentant un effet anti-hypertenseur, a montré qu’il est très sensible à l’hydrolyse et à l’oxydation. Sept produits de dégradation ont été identifiés pour ce produit dont un schéma probable de fragmentation a été établi. Des diagrammes de phases pression-température ont été construits pour le bicalutamide et le finastéride, médicaments du cancer de prostate, en utilisant une approche topologique basée simplement sur les données disponibles dans la littérature. Cette étude a montré que la relation thermodynamique (énantiotropie ou monotropie) entre les formes cristallines sous conditions ordinaires peut être modifiée en fonction de la température et de la pression. Ce résultat est important pour la production des médicaments car il montre comment une telle information peut être obtenue par des mesures simples et accessibles aux laboratoires de recherche industrielle, sans que ces derniers soient contraints d’expérimenter sous pression. La méthode topologique de construction de diagramme de phases a été validée ensuite en la comparant à une méthode expérimentale consistant à suivre, par analyse thermique, des transitions de phases en fonction de la pression. La méthode expérimentale a été appliquée à deux composés, la benzocaine, anesthésique local, et le chlorhydrate de cystéamine, médicament utilisé pour les cystinoses. Les deux formes étudiées de benzocaine présentent une relation énantiotrope qui se transforme en relation monotrope à haute pression. Une nouvelle forme cristalline (forme III) du chlorhydrate de cystéamine a été découverte au cours de ce travail. La relation thermodynamique entre cette forme III et la forme I est énantiotrope dans tout le domaine de température et de pression. De plus, le chlorhydrate de cystéamine, classé hygroscopique, a fait l’objet d’une étude quantitative de sa sensibilité à l’eau, montrant qu’il devient déliquescent sans formation préalable d’hydrate (...) / The development of a drug for a given therapeutic target requires several steps, which can be summarized by drug screening, a preclinical phase and a number of clinical phases. These steps allow the selection of an active substance and a verification of its therapeutic efficacy and toxicological safety. The latter two criteria define the quality of the drug, which once demonstrated, must be guaranteed throughout its shelf life. Quality is assessed through stability studies that are carried out with the raw material of the active substance (preformulation phase) and with the final product. The intrinsic stability of the active substance depends on its chemical and physical properties and their characterization is the core of the stability studies, which in addition consists of sensitivity studies of the active pharmaceutical ingredient (API) for environmental factors that can modify the intrinsic properties of the substance. The approach presented in this work is based on the one hand on the assessment of the chemical stability, i.e. the reactivity of APIs through chemical purity studies and forced degradation in solution, and on the other hand on the assessment of the physical stability. For the latter, crystalline polymorphism is of great importance, as is the ability of the API to form hydrates or solvates. The study of crystalline polymorphism is based on the construction of pressure-temperature phase diagrams in accordance with thermodynamic requirements leading to the stability condition domains of the different crystalline forms. The stability behavior of five APIs used or meant for oral applications has been studied as part of this work. The chemical analysis of tienoxolol, an antihypertensive drug, has demonstrated its sensitivity for hydrolysis and oxidation. Seven degradation products were identified and patterns of fragmentation have been established. Pressure-temperature phase diagrams have been constructed for bicalutamide and finasteride, drugs against prostate cancer, using a topological approach based on data available in the literature. The study demonstrates that the thermodynamic relationship (enantiotropy or monotropy) between crystalline forms under ordinary conditions can change depending on the pressure. This is important for drug development as it demonstrates how stability information can be obtained by standard laboratory measurements accessible to industrial research laboratories without the necessity to carry out experiments under pressure. The topological approach for the construction of phase diagrams has subsequently been validated by measuring transition temperatures as a function of pressure. Experiments have been carried out with benzocaine, a local anesthetic, and with cysteamine hydrochloride, a drug used against cystinosis. Two crystalline forms were observed in the case of benzocaine. They exhibit an enantiotropic relationship that becomes monotropic at high pressure. For cysteamine hydrochloride, a new crystalline form (form III) was discovered. The thermodynamic relationship between the new form III and the known form I is enantiotropic for the entire temperature and pressure range. Cysteamine hydrochloride’s sensitivity to water has been studied, as it is hygroscopic. It has been demonstrated that it becomes deliquescent in the presence of water and no trace of a hydrate has been found. Finally, a study combining thermal and chromatographic methods showed that, under the effect of temperature, cysteamine hydrochloride turns into cystamine in the solid as well as in the liquid state, The latter is known to be an important impurity of cysteamine hydrochloride. In conclusion, the approach developed in this work allowed to characterize the stability properties of a number of APIs and to determine the factors that may change these properties and influence the intrinsic stability (...)