During and after medical treatment, pharmaceutical compounds as well as their metabolites and conjugates are excreted from the users through urine and feces. The pharmaceuticals end up in wastewater treatment plants, which are not designed to deal with this kind of organic micro-pollutant. Eventually the pharmaceuticals end up in the environment where they can have adverse physiological and behavioral effects on aquatic life and could contribute to the spread of antibiotic resistance among microorganisms. Adsorption to activated carbon is an established method for removal of pharmaceuticals from wastewater. It is however quite expensive and it is of interest to identify cost-effective alternatives. One possible alternative is bark, which is a common by-product from forest industry and has a complex microstructure and high porosity compared to many other naturally occurring materials. In order to investigate the potential of using bark to remove pharmaceuticals from municipal wastewater four column filters were built, two with activated carbon and two with bark. They were used in an experiment conducted at Kungsängsverket, the largest wastewater treatment plant in Uppsala municipality. The objectives were to assess pharmaceutical concentrations in treated wastewater at Kungsängsverket and to compare the performance of bark and activated carbon filters under different loading rates. During this time the filters were run at different loading rates and two different types of bark was used. 24 common pharmaceuticals from different therapeutic groups were targeted. The pharmaceutical concentrations measured at Kungsängsverket were generally low, but mean concentrations of five pharmaceuticals (atenolol, metoprolol, furosemide, hydrochlorothizide and diclofenac) exceeded 250 ng/l. Out of these, four have been shown to have adverse effects on aquatic life and it would be preferable if they were not released into the recipient. Bark was not as good at removing pharmaceuticals from wastewater as activated carbon was, but decent removal rates were achieved for several compounds. The removal rates of either filter type did not seem to be significantly impacted by variations in loading rate or bark size. The concentrations of a few compounds increased after treatment with the bark filters and the reason for this is not clear. One possibility is interference from other organic substances in the wastewater or the bark, but determining the reason for this increase should be a priority for any further research on the subject. Another problem encountered during the project that is likely to pose a problem for future implementation is that the bark filters were very sensitive to clogging. Running the filters at full scale would require frequent back-washing which would be a disadvantage from both economical and practical reasons.

The occurrence of pharmaceuticals in environment originating from human consumption has received increased scientific attention during the last decades due to concerns regarding their combined environmental effects in aquatic and terrestrial environments, in flora and biota and by extent in human health. In this review, I summarized the existing knowledge on the entire life cycle of pharmaceutical substances, from their exposure (sources) and fate to their effects on the natural environment. Since the negative effects of several drugs along with the environmental damage they entail are now known, it can be suggested that pharmaceutical companies make greener pharmaceutical products to reduce these effects to the terrestrial and aquatic environment. The present review could provide suggestions to improve the pharmaceutical environmental management globally, such as methodologies for monitoring systems, that need to be put in place for consistent data collection. Another area of research that is important is the release of pharmaceutical compounds in manufacturing plants as well as from landfill effluent. Finally, one more area with need for further research is green chemistry which could reduce or even eliminate the potential hazards of pharmaceutical compounds that enter the environment, irrespective to the source of entry.

5.Abstract Title: Legal aspects of a parallel trade in drugs within the EU Key words: single market, parallel trade, price regulation, pharmaceuticals Parallel trade is one of the most distinctive benefits of the European single market. In a simplified way, parallel trade is legal importing of drug from one country where it is legally marketed into a second country where the marketing authorization holder also markets that product, but without the consent of the marketing authorization holder. Price regulation of medical products makes parallel trade of drugs questionable. Final price for customer is not result of an independent decision of pharmaceutical company but it is given by relevant authority often regarding the spending power of citizens. Wholesalers are taking advantage of these price differences when they are buying cheap drugs in low-price countries and selling them with profit in countries where drug price are higher. In theory this practice should deliver significant saving for patient and national healthcare insurance system in destination countries due to lower price of parallel traded drugs. However many studies show minimum impact on drug price in these countries since wholesalers retain great part of the financial benefits. On the other hand, negatives of parallel trade are evident. Drug...

The thesis reviews current patent strategies of original pharmaceutical companies and their tangled role in the fabric of European pharmaceutical innovation and competition. It addresses several components of the European pharmaceutical industry such as regulatory framework, patent filing and dispute strategies and competition law. It argues that patent law is embedded in a broader competition law framework however plays on a separate field where it governs primarily the entry to its exclusive space by market actors. However it asserts that competition law should serve as a time referee for the patent law playfield and check if the abusive prolongation of exclusive patent position does not occur. The thesis deliberates that in view of ever rising number of patent applications, abuse of the patent system may become symptomatic to the system. The Commission data presented in the Final Report on the pharmaceutical sector inquiry are again inspected. Although data should be used with caution, it revealed a good quantitative base for assessment of a system which seemingly becomes more entropic, complex and susceptible to abuse. Therefore the underlying principles in both patent and competition law should be upheld more strongly than ever. It is the principle of fairness that should have normative force...

Pharmaceuticals are group of organic substances with significant worldwide consumption in human and veterinary medicine. These compounds may be metabolized in the organism, but in some cases they remain unchanged and both are usually excreted via renal excretion in the native form or as metabolites. Large quantities of pharmaceuticals and their metabolites contaminate municipal wastewater. The wastewater treatment plants are unable to remove these substances completely, so they contaminate surface water, groundwater and soil as well. Due to the biological activity of pharmaceuticals, long - term effect may cause bacterial resistance, endocrine influence, DNA and renal damages in non-target organisms. The phytoextraction and the translocation of radiolabeled diclofenac with 125 I were experimentally studied by using of in vitro cultivated plants Helianthus annuus and Zea mays. Efficiency od phytoextraction was monitored as decrease of radioactivity of tested substance [125 I]diclofenac in Murashige-Skoog cultivation medium. Both species are able to extract tested substance during 8 to 10 days of cultivation, with efficiency approximately 85 % using Zea mays and 79 % using Helianthus annuus. Better extraction ability of diclofenac was observed at Helianthus annuus - 80 mg/ kg of dry weight compared...

Cette thèse de doctorat porte sur la compréhension des aspects concurrentiels du marché de l’innovation incrémentale en France en s’appuyant sur une approche empirique. A cette fin, l’analyse porte plus particulièrement sur l’innovation incrémentale et les dynamiques concurrentielles des médicaments appelés en France « similaires ». La littérature anglophone retient les expressions « me-too » ou « follow-on » pour définir les produits pharmaceutiques à faible valeur ajoutée et qui ont des caractéristiques anatomiques, thérapeutiques et chimiques proches des molécules précédemment mises sur le marché. Le premier chapitre vise notamment l’étude sur les parts de marché des médicaments « follow-on » et le lien avec la variable stratégique définie par l’ordre d’arrivée sur le marché. Dans ce chapitre de thèse, on vérifie que les premiers follow-on d’une classe thérapeutique ont un avantage concurrentiel en ce qui concerne la capacité des capturer et de maintenir des parts de marché pendant une longue période. En outre, les parts de marché sont positivement corrélés avec l’habilité de la firme de fixer un prix supérieur au premier médicament de la classe. Cet avantage stratégique des premiers entrants est aussi intrinsèquement lié aux caractéristiques qualitatives des produits puisque nous avons pu constater que les parts de marché des médicaments sont directement corrélés avec le niveau d’innovation du produit. Le deuxième chapitre de la thèse apporte des éléments sur la nature concurrentielle en termes de prix du marché de l’innovation pharmaceutique incrémentale. Deux variables dépendantes mesurant le prix ont été considérées : le prix du produit calculé en coût du traitement journalier et le prix du médicament divisé par la moyenne des prix des médicaments similaires dans la classe. Nous vérifions que les derniers entrants ont une tendance à avoir un prix inférieur aux premières molécules « follow-on ». Cela implique une caractérisation des derniers entrants ayant un moindre pouvoir de négociation avec le régulateur autour du prix, notamment en raison d’une qualité innovatrice intrinsèque plus faible. Le troisième chapitre présente une analyse empirique autour de la confrontation des deux marchés émergents et d’importance majeure pour la régulation des systèmes de santé : le marché des génériques et le marché des médicaments « follow-on ». L’analyse se concentre sur les aspects de l’intensité de la compétition dans le marché des médicaments brevetés similaires et son impact sur la pénétration des versions génériques de ces derniers. Le constat est que l’intensité de la compétition de médicaments similaires est positivement et significativement corrélée avec l’introduction des génériques. Plus de produits « follow-on » entrainerait donc une baisse encore plus importante des prix des génériques des derniers « follow-on » dans la classe thérapeutique. L’approche essentiellement empirique de cette thèse doctorale permet ainsi de mieux comprendre les déterminants et la dynamique de ce marché relativement émergent et qui suscite des nombreux débats au sein de la communauté scientifique. Enfin nous terminons par une brève conclusion générale fondé autour des résultats de cette recherche permettent d’affirmer que dans un marché régulé comme celui qui prévaut en France, l’intensité de la compétition, engendré notamment par l’arrivé sur le marché des médicaments « follow-on », peut avoir des conséquences positives sur les aspects concurrentiels du secteur du médicament...... / The dynamics of pharmaceutical markets have been constantly changing last years. The development of the so called “follow-on” or “me-too” drugs has been in the centre of a major debate concerning the ability of innovation in the health sector. These drugs are characterized by having a minor level of innovation and do not add any therapeutic value in relation to the previous drugs launched in the market. This doctoral dissertation proposes three empirical essays concerning competition aspects in the market of incremental innovation in France. The first chapter focuses on the impact of entry order on “follow-on” drugs competition in the French market between years 2001 and 2007. More precisely, this study examines the effects on market share of first entrants in the follow-on drug market and how this possible competitive advantage changes over time. Our results are coherent with theoretical microeconomic issues concerning the importance of being first. We find evidence that first movers in the follow on drug market have the ability to capture and maintain greater market share for a long period of time. The hierarchical market position of follow on drugs does not seem to be affected by generic drugs emergence. From a dynamic perspective, our analysis shows that market share is positively correlated with the ability of follow on drugs to set prices higher than the average follow-on drug price in a specific therapeutic class (ATC) which means that market power remains considerably important for first movers. Finally we found that the optimum level of innovation to maximize market share is the highest one.The second chapter examines the relationship between entry order of follow-on drugs and their prices on the French pharmaceutical market. We used a representative panel data of 1047 follow-on drug formulations distributed in 118 ATC classes set over 2001-2007. Two measures of prices were used in the econometric specifications: the absolute price and the relative price of the follow-on drugs. The former concerns simply the absolute price of the drug in daily costs and the latter is the price of the drug relative to the average price of the follow-on drugs in the class. Both prices are calculated on the basis of manufacturer prices. These different indicators give us similar results for the impact of entry order on prices but they are differently correlated with market share. Moreover, our study analyses the potential impact of several variables on prices of pharmaceutical incremental innovation such as firm size, innovation and intensity of competition. Our results suggest that big firms have more ability to negotiate higher prices and that the number of follow-on drugs in the class and the emergence of generic competition may help decrease general prices in the ATC class. We have not found any relationship between prices and innovation in the French pharmaceutical market.The third chapter investigates the potential relationship between follow-on drugs dissemination and generic drug market emergence. It explores the structural determinants of off-patented drugs development at the therapeutic class level with a focus on explanatory variables that reflect the intensity of competition amongst similar interchangeable drugs. We found that generic market growth is greater in therapeutic classes where the number of similar drugs is high and the average brand price is low. This could be due to the fact that brand name drugs reduce their prices to keep market share when generic drugs enter the market. We study also the generic to follow-on brand price ratio at the individual drug level and we found that generic prices of later follow-on drugs are closer to the price of the brand name than generics of first follow-on movers. Our results are coherent with the fact that intensity of competition in the follow-on drugs may help reduce prices not only in the patented drug markets but also in the off-patented sector.

Freeze-drying of protein pharmaceuticals is a procedure frequently used to obtain stability of the active pharmaceutical ingredientduring distribution and storage. It can be performed in pre-filled syringes, with a lubricous coating of silicone on the inside, to enable the piston moving. The coating changes the environment potentially affecting the features of the freeze-dried cake since the wetting behavior and adhesion to the inner wall is affected.This project aimed to investigate the effect of the siliconization of the cakes. Three different formulations were freeze-dried in nonsiliconized (NS) and siliconized vials using different siliconization protocols. Analysis was done using differential scanning calorimetry (DSC), thermal gravimetric analysis (TGA),scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and an embedding method, intended to give information about the cake’s shrinkage, cracking and pore-structure. The water content in the bottom of the cakes was consistently higher than in the top, a difference decreasing over time. Increased surface hydrophobicity lead to increased shrinkage of the cake’s volume and a decrease in fogging. The bottom of the protein cake in the vial siliconized with a commercial silicone emulsion consisted of pores with regularly equal pore size and thick pore walls, a structure not seen in any other cake. All cakes in the silicone emulsion siliconized vials had lower water content than the cakes in the vials using the other siliconization method. The XPS-analysis showed that the cakes in the emulsion siliconized vials contained silicon, indicating an excess of silicone when siliconizing and/or an unstable coating. / NextBioForm

O emprego da catálise de transferência de fase substituindo uma ou mais etapas da síntese de um fármaco, pode ser um recurso vantajoso sob o ponto de vista tecnológico. Empregado na síntese de importantes fármacos que são a dicicloverina, caramifeno eamiodarona, a catálise de transferência de fase revelou-se muito vantajosa na síntese dos dois primeiros. A dicicloverina e o caramifeno tiveram as suas rotas de obtenção verticalizadas, com bons resultados e custos bem mais baixos quando comparados com os atualmente adquiridos no exterior. Ficou evidente que a catálise de transferência de fase pode ser aplicada com sucesso na produção destes e outros fármacos em associação com técnicas já conhecidas com ótimos resultados. / The phase transfer catalysis application replacing one or more steps in the drug synthesis, can be of advantage in the technologycal approach. Applied to the synthesis of important drugs such as diciclomine, caramiphen,and amiodarone, the phase transfer catalysis resulted in a very advantageous method in the first two drugs. The diciclomine and the caramiphen, gets their obtention routes from raw materiaIs wi th good resul ts and low coasts, when compared wi th the drugs presently purchassed in foreign countries. It is clear that the phase transfer catalysis, can be applied succesfully in the manufacturing of these and other drugs, in conjuction with known techniques with good results.

Triclosan é um composto orgânico de baixa solubilidade que vem sendo utilizado em formulações de cremes dentais e faciais, xampu, sabonetes, embalagens de gêneros alimentícios e diversos tipos de materiais, tais como, adesivos, brinquedos, sapatos, selantes, tintas, colchão, roupas, pisos, toldos e rejuntes. O amplo uso deste composto deve-se à grande eficácia contra bactérias Gram negativas e Gram positivas. Por seu extenso uso, evidências da presença de Triclosan têm sido frequentemente relatadas em efluentes urbanos e industriais, águas superficiais e sedimentos de ambientes dulcícolas, estuarinos e marinhos, como também em organismos aquáticos como algas, peixes e mamíferos. Neste contexto, o presente estudo avaliou a toxicidade aguda e crônica de Triclosan para diferentes invertebrados marinhos de águas tropicais. Para tanto, ensaios de toxicidade aguda foram realizados com o copépodo Nitokra sp (mortalidade) e com o ouriço-do-mar Lytechinus variegatus (taxa de fertilização). Para a avaliação do efeito crônico, ensaios de toxicidade de curta duração (desenvolvimento embriolarval) foram realizados com o ouriço-do-mar L. variegatus e Perna perna. Além desses métodos, o ensaio do Tempo de Retenção do Corante Vermelho Neutro foi empregado com a finalidade de se avaliar os efeitos do Triclosan sobre a estabilidade da membrana lisossômica de hemócitos de P. perna. Na avaliação do efeito agudo, o valor médio da CL(I)50;96h encontrada para o copépodo foi de 0,20 mg.L-1 enquanto que o valor médio da CI(I)50;1h para ouriço-do-mar foi de 0,28 mg.L-1. Já na avaliação do efeito crônico, o valor médio da CI(I)50;24h para ouriço-do-mar foi de 0,14 mg.L-1 e para o molusco bivalve a média da CI(I)50;48h, foi de 0,13 mg.L-1. O efeito na estabilidade da membrana lisossômica de hemócitos de P. perna ocorreu em concentrações a partir de 12 ng.L-1. Estes resultados evidenciam o risco ecológico da introdução contínua desse composto em ambientes marinhos, e devem ser considerados para identificação de concentrações seguras e futura regulação do bactericida Triclosan na legislação ambiental nacional e internacional. / Triclosan is a low solubility organic compound that has been used in toothpastes, face cream, shampoos, soaps, food packages, and a variety of other materials such as stickers, toys, shoes, paints, clothes, tiles, awnings and grout. The reason for its intense use as biocide is its great efficacy against Gram-negative and Grampositive bacteria. Evidences of Triclosan presence in urban and industrial effluents, superficial waters and sediments from freshwater, estuarine, and marine environments, as well as aquatic organisms (algae, fishes, mammals) have been reported in the literature. In this context, the present study assessed the acute and chronic toxicity of Triclosan to different tropical marine invertebrates. Acute toxicity bioassays using the copepod Nitokra sp (mortality) and the sea-urchin Lytechinus variegatus (fertilization rate) were performed. Short-term chronic toxicity bioassays with Lytechinus variegatus and the bivalve mussel Perna perna were carried out in order to assess Triclosan chronic effects. Besides, the Neutral Red Retention Time assay was employed to evaluate the effect of Triclosan on the stability of lysosomal membrane of hemocytes of Perna perna. In the acute toxicity assays, the mean value of LC(I)50;96h obtained for the copepod was 0.20 mg L-1, whereas the mean value of IC(I)50;1h for the sea-urchin was 0.28 mg L-1. In the chronic toxicity assays, the mean value of IC(I)50;24h recorded for the seaurchin was 0.14 mg L-1, whilst for the bivalve mollusk the mean value of IC(I)50;48h was 0.13 mg L-1. The effect on the lysosomal membrane stability of Perna perna hemocytes started to occur from 12 ng L-1. The results evidence the ecological risk associated to the continuous introduction of Triclosan into marine aquatic environments and must be considered in the identification of safety concentrations and future regulation of this bactericide compound in national and international environmental legislation.

A crescente demanda por produtos farmacêuticos tem como consequência a necessidade de elaboração de novos procedimentos analíticos que, via de regra, apresentem características como: elevada frequência analítica, simplicidade, ausência de substâncias altamente tóxicas, e com baixo custo para o controle de qualidade dos fármacos lançados no mercado. Neste trabalho descreve-se o desenvolvimento de procedimentos para determinação de N-acetilcisteína (NAC), nimesulida (NMS), trimetoprima (TMP) e sulfametoxazol (SMX) em formulações farmacêuticas usando técnicas eletroanalíticas e métodos de separação. Na determinação da NAC foi usado um eletrodo de grafite pirolítico modificado com ftalocianina de cobalto (CoPc) em 0,1 mol L-1 de NaOH. Este sensor mostrou intensa catálise para NAC produzindo um aumento na corrente anódica de cerca de 700 vezes quando comparado com o eletrodo limpo. Um procedimento em FIA foi proposto usando este sensor, e a curva analítica foi linear no intervalo 5,0 × 10-5 a 1,0 × 10-3 mol L-1, com coeficiente de correlação maior que 0,999. O DPR de 1,1% (n = 13) foi obtido para uma solução contendo 5,0 × 10-4 mol L-1 de NAC. Além disso o método apresentou LD e LQ de 9,0 ×10-7 mol L-1 e 3,0 × 10-6 mol L-1, respectivamente. A determinação eletroanalítica de NMS foi conduzida em solução de 1,0 mol L-1 de NaOH usando eletrodo de carbono vítreo, meio em que não ocorre envenenamento da superfície do eletrodo. Desse modo, um método eletroanalítico usando voltametria de onda quadrada foi elaborado para análise de NMS. Nas condições ótimas de análise o método apresentou uma faixa linear no intervalo de 4,3 × 10-5 a 4,2 × 10-4 mol L-1, com limite de detecção de 3,2 × 10-6 mol L-1. A precisão do método foi avaliada em um mesmo dia de análise por sucessivas medidas voltamétricas (n = 10) em solução de hidróxido de sódio 1,0 mol L-1 contendo 1,0 × 10-4 mol L-1 de NMS, sem a necessidade de renovação da superfície do eletrodo. Os resultados para esta série de medidas foram avaliados e o DPR calculado foi somente 1,2%, indicando que nessas condições o eletrodo apresentou uma resposta estável além de não sofrer qualquer efeito de envenenamento. Para a determinação simultânea de TMP e SMX foi desenvolvido um método analítico empregando eletroforese capilar com detecção condutométrica sem contato (C4D). Usando uma solução 10 mmol L-1 lítio-fosfato (pH 7,1 ) como tampão de corrida, a faixa linear de trabalho obtida para ambos os analitos foi de 12,5 a 200 × 10-6 mol L-1 com R > 0,999. Nas condições de análise foram obtidos LDs de 1,1 × 10-6 mol L-1 para a TMP, e 3,3 × 10-6 mol L-1 para o SMX com um tempo total de análise de 2,6 minutos. / The growing demand for pharmaceutical products has created need for developing new analytical procedures that, as a rule, exhibit characteristics like, high sampling rate, simplicity, absence of highly toxic substances, and low cost for quality control of drugs launched in market. In this work, electroanalytical and capillary electrophoresis methods for determination of N-acetylcysteine (NAC), nimesulide (NMS), trimethoprim (TMP) and sulfamethoxazole (SMX) in pharmaceutical formulations were proposed. For NAC determinations a pyrolytic graphite electrode modified with CoPc in 0.1 mol L-1 NaOH was used. This new sensor showed to be catalytic for this compound producing an anodic current increase of 700 times when compared with bare electrode. A FIA procedure was proposed using this new sensor, and the method showed a linear response in a range of 5.0 × 10-5 to 1.0 × 10-3 mol L-1 with correlation coefficient better than 0.999. RSD of 1.1% (n = 13) was obtained for a solution containing 5.0 × 10-4 mol L-1 NAC. Furthermore the method presented DL and QL of 9.0 × 10-7 mol L-1 and 3.0 × 10-6 mol L-1, respectively. NMS electroanalytical determination was conducted in a 1.0 mol L-1 NaOH solution using a glassy carbon electrode. In strongly alkaline medium it was verified that NMS does not produce poisoning of the electrode surface. Thus an electroanalytical method using square wave voltammetry was proposed for analysis of NMS. In the optimal analysis conditions proposed method showed a linear response in a range of 4.3 × 10-5 to 4.2 × 10-4 mol L-1, with detection limit of 3.2 × 10-6 mol L-1. Accuracy of the method was evaluated on the same day by successive voltammetric measurements (n = 10) in a 1.0 mol L-1 sodium hydroxide solution containing 1.0 × 10-4 mol L-1 of NMS without the needing to renew electrode surface. Results for this series of measures were calculated and standard deviation was of only 1.2%, suggesting that in these conditions the electrode showed a stable response in addition to suffer no poisoning effects. For simultaneous determination of TMP and SMX an analytical method using capillary electrophoresis with contactless conductivity detection (C4D) was developed. Using a 10 mmol L-1 lithium-phosphate (pH 7.1) solution as running buffer, a linear response range for both analytes was obtained ranging through 12.5 to 200 × 10-6 mol L-1 with R> 0.999. Under these analysis conditions, DL of 1.1 × 10-6 mol L-1 for TMP, and 3.3 × 10-6 mol L-1 for SMX were obtained, with a total analysis time of only 2.6 minutes.