Made available in DSpace on 2014-12-17T14:16:26Z (GMT). No. of bitstreams: 1 EdilenePL_DISSERT.pdf: 1151865 bytes, checksum: 7a412187753da50d4030404745c567f7 (MD5) Previous issue date: 2010-02-26 / According to the global framework regarding new cases of tuberculosis, Brazil appears at the 18th place. Thus, the Ministry of Health has defined this disease as a priority in the governmental policies. As a consequence, studies concerning treatment and prevention have increased. Fixed-dose combination formulations (FDC) are recognized as beneficial and are recommended by WHO, but they present instability and loss on rifampicin bioavailability. The main purpose of this work was to carry out a pre-formulation study with the schedule 1 tuberculosis treatment drugs: rifampicin, isoniazid, pyrazinamide and ethambutol and pharmaceutical excipients (lactose, cellulose, magnesium stearate and talc), in order to develop an FDC product (150 mg of rifampicin + 75 mg of isoniazid + 400 mg of pyrazinamide + 250 mg of ethambutol). The studies consisted of the determination of particle size and distribution (Ferret s diameter) and shape through optical microscopy, as well as rheological and technological properties (bulk and tapped densities, Hausner Factor, Carr s Index, repose angle and flux rate) and interactions among drugs and drug excipient through thermal analysis (DSC, DTA, TG and your derivate). The results showed that, except isoniazid, the other drugs presented poor rheological properties, determined by the physical characteristics of the particles: small size and rod like particles shape for rifampicin; rectangular shape for pyrazinamide and ethambutol, beyond its low density. The 4 drug mixture also not presented flowability, particularly that one containing drug quantity indicated for the formulation of FDC products. In this mixture, isoniazid, that has the best flowability, was added in a lower concentration. The addition of microcrystalline cellulose, magnesium stearate and talc to the drug mixtures improved flowability properties. In DSC analysis probable interactions among drugs were found, supporting the hypothesis of ethambutol and pyrazinamide catalysis of the rifampicin-isoniazid reaction resulting in 3- formylrifamycin isonicotinyl hydrazone (HYD) as a degradation product. In the mixtures containing lactose Supertab? DSC curves evidenced incompatibility among drugs and excipient. In the DSC curves of mixtures containing cellulose MC101?, magnesium stearate and talc, no alterations were observed comparing to the drug profiles. The TG/DTG of the binary and ternary mixtures curves showed different thermogravimetrics profiles relating that observed to the drug isolated, with the thermal decomposition early supporting the evidences of incompatibilities showed in the DSC and DTA curves / De acordo com o quadro mundial da tuberculose, o Brasil ocupa o 18? lugar em n?mero de casos novos, assim o Minist?rio da Sa?de definiu a doen?a como prioridade entre as Pol?ticas Governamentais de Sa?de. Desde ent?o se intensificaram os estudos relacionados ao tratamento e preven??o desta doen?a. As formula??es em dose fixa combinada (DFC) s?o reconhecidas como ben?ficas e apoiadas pela OMS, mas apresentam problemas de instabilidade e queda na biodisponibilidade da rifampicina. O objetivo principal desse trabalho foi realizar estudo de pr?-formula??o com os f?rmacos que integram o esquema 1 para o tratamento da tuberculose: rifampicina, isoniazida, pirazinamida e etambutol e excipientes farmac?uticos (lactose, celulose, estearato de magn?sio e talco), visando o desenvolvimento de um produto em dose fixa combinada (150 mg de rifampicina + 75 mg de isoniazida + 400 mg de pirazinamida + 250 mg de etambutol). Os estudos consistiram na determina??o do tamanho e distribui??o das part?culas (di?metro de Ferret) e forma por microscopia ?ptica, al?m das propriedades reol?gicas e tecnol?gicas (densidades aparente e de compacta??o, Fator de Hausner, ?ndice de Carr, ?ngulo de repouso e velocidade de escoamento) e das intera??es entre os f?rmacos e f?rmacos-excipientes por an?lise t?rmica (DSC, DTA, TG e sua derivada). Os resultados mostraram que ? exce??o da isoniazida, os demais f?rmacos apresentaram propriedades reol?gicas pobres, determinadas pelas caracter?sticas f?sicas das part?culas: tamanho reduzido e forma de agulhas da rifampicina; forma retangular da pirazinamida e etambutol, al?m da baixa densidade deste ?ltimo. A mistura dos quatro ativos tamb?m n?o apresentou fluxo, especialmente a prepara??o contendo a quantidade de f?rmacos preconizada para a formula??o de produtos em dose fixa combinada, uma vez que nessa mistura, a isoniazida, que possui o melhor fluxo, foi adicionada a uma concentra??o menor. A adi??o de celulose microcristalina, estearato de magn?sio e talco ?s misturas dos f?rmacos, melhorou as propriedades de fluxo. Nas an?lises por DSC foram encontradas prov?veis intera??es entre as subst?ncias ativas, refor?ando a hip?tese de cat?lise por etambutol e pirazinamida para a rea??o entre rifampicina e isoniazida que resulta no produto de degrada??o 3-(isonicotinoilhidrazinometil)rifamicina. Nas curvas de DSC das misturas contendo lactose Supertab? foi evidenciada a ocorr?ncia de incompatibilidade entre os f?rmacos e o excipiente. As curvas de DSC das misturas contendo celulose MC101?, estearato de magn?sio e talco n?o apresentaram altera??es em rela??o ao perfil dos f?rmacos. As curvas de TG/DTG das misturas bin?rias e tern?rias apresentaram perfis termogravim?tricos diferentes em rela??o ao observado para os f?rmacos isoladamente, com in?cio da decomposi??o t?rmica antecipada, dando suporte as evid?ncias de incompatibilidades encontradas nas curvas de DSC e DTA

Tuberculosis is an ancient infectious disease and a leading cause of death globally. Preclinical research is important for defining drugs and regimens which should be carried forward to human studies. This thesis aims to characterize the population pharmacokinetics and exposure-response relationships of anti-tubercular drugs alone and in combinations, and to suggest experimental designs for preclinical settings. The population pharmacokinetics of rifampicin, isoniazid, ethambutol and pyrazinamide were described for the first time in two mouse models. This allowed for linking the population pharmacokinetic model to the Multistate Tuberculosis Pharmacometric (MTP) model for biomarker response, which was used to characterize exposure-response relationships in monotherapy. Pharmacodynamic interactions in combination therapies were quantitatively described by linking the MTP model to the General Pharmacodynamic Interaction (GPDI) model, which provided estimates of single drug effects together with a quantitative model-based evaluation framework for evaluation of pharmacodynamic interactions among drugs in combinations. Synergism (more than expected additivity) was characterized between rifampicin and ethambutol, while antagonism (less than expected additivity) was characterized between rifampicin and isoniazid in combination therapies. The new single-dose pharmacokinetic design with enrichened individual sampling was more informative than the original design, in which only one sample was taken from each mouse in the pharmacokinetic studies. The new oral zipper design allows for informative pharmacokinetic sampling in a multiple-dose administration scenario for characterizing pharmacokinetic-pharmacodynamic relationships, with similar or lower bias and imprecision in parameter estimates and with a decreased total number of animals required by up to 7-fold compared to the original design. The optimized design for assessing pharmacodynamic interactions in the combination therapies, which was based on EC20, EC50 and EC80 of the single drug, provided lower bias and imprecision than a conventional reduced four-by-four microdilution checkerboard design at the same total number of samples required, which followed the 3Rs of animal welfare. In summary, in this thesis the population pharmacokinetic-pharmacodynamic models of first-line drugs in mice were characterized through linking each population pharmacokinetic model to the MTP model. Pharmacodynamic interactions were quantitatively illustrated by the MTP-GPDI model. Lastly, experimental designs were optimized and recommended to both pharmacokinetic and pharmacodynamic studies for preclinical settings.

FundaÃÃo de Amparo à Pesquisa do Estado do Cearà / Baixas concentraÃÃes sÃricas de drogas antituberculose podem ser causa da resistÃncia de Mycobacterium tuberculosis Ãs drogas utilizadas para tratamento, a qual à mais freqÃente em pacientes em tratamento irregular, mas pode ser verificada em pacientes em tratamento diretamente observado. Os objetivos desse estudo foram avaliar a permeabilidade intestinal e a biodisponibilidade de rifampicina, isoniazida e pirazinamida em pacientes com tuberculose pulmonar ativa. Realizou-se estudo transversal controlado. No perÃodo entre julho de 2004 e dezembro de 2005 foram selecionados 56 pacientes consecutivos com tuberculose pulmonar ativa, atendidos no Centro de SaÃde Carlos Ribeiro em Fortaleza, Cearà e 29 controles sadios recrutados entre profissionais de saÃde, seus familiares e vizinhos para avaliaÃÃo da permeabilidade intestinal. Foram coletadas informaÃÃes sociodemogrÃficas e exames bioquÃmicos e realizou-se o teste de lactulose/manitol de todos. Os primeiros 30 casos de tuberculose e os 29 controles dessa amostra foram selecionados para a avaliaÃÃo da biodisponibilidade sÃrica de rifampicina, isoniazida e pirazinamida em amostras coletadas duas horas e seis horas depois da ingestÃo. A mÃdia de idade dos casos foi de 39,2  15 anos e a dos controles 34  11,0 (p=0,61). Eram do sexo masculino 71 % dos casos e 66% dos controles (p=0,57). O peso mÃdio dos casos (52,4  6,3 kg) foi significativamente menor do que o dos controles (71,1  14,0 kg) (p=0,014). Verificou-se menor excreÃÃo de lactulose entre os casos (mediana de 0,1721%; variando de 0,0-5.0139%) do que entre os controles (0,4301%; variando de 0,0-2,064) (p=0,049%); a excreÃÃo de manitol entre os casos foi 17,9910% (1,9567-71,5446%) e entre controles foi de 24,3899% (1,3883-63,0539%) (p=0,147); a relaÃÃo lactulose/manitol foi de 0,0095 (0,0-0,0759%) entre os casos e 0,0136 (0,0-0,0136%) entre os controles (p=0,018). A concentraÃÃo sÃrica mÃxima de rifampicina teve mÃdia de 1,46  0,72 Âg/ml nos casos e de 6,69  3,07 Âg/ml nos controles (p<0,001); a de isoniazida foi 2,62  1,53 Âg/ml entre os casos e 1,98  0,76 Âg/ml entre os controles (p=0,057) e a de pirazinamda foi de 44,10  10,40 Âg/ml entre os casos e 36,32  12,02 Âg/ml entre os controles (p=0,007). Quatro (13,3%) casos nÃo chegaram a alcanÃar os limites mÃnimos das concentraÃÃes normais esperadas de nenhuma das drogas de primeira linha para o tratamento da tuberculose; 21 (70,0%) alcanÃaram essa concentraÃÃo sÃrica apenas para uma droga (pirazinamida) e cinco (16,7%) apenas para duas drogas (pirazinamida e isoniazida). Nenhum caso alcanÃou as concentraÃÃes sÃricas normais esperadas para as trÃs drogas, simultaneamente. Em conclusÃo, observou-se reduÃÃo da absorÃÃo paracelular entre os pacientes com tuberculose bem como mà absorÃÃo intestinal de rifampicina e isoniazida. Estes resultados sugerem a necessidade da avaliaÃÃo de medidas para reduzir a mà absorÃÃo intestinal de drogas e evitar o retardo ou a impossibilidade de cura da doenÃa, bem como o risco de multirresistÃncia / Reduced antituberculosis drugs concentrations are associated with Mycobacterium tuberculosis resistance, mostly in patients in irregular but also in directly observed treatment. This study aims to evaluate intestinal permeability and bioavailability of rifampin (R), isoniazid (I) and pyrazinamide (P) in patients with active pulmonary tuberculosis. A controlled cross sectional study evaluated intestinal permeability from 56 consecutive active pulmonary tuberculosis (TB) patients who attended Carlos Ribeiro Health Unit in Fortaleza, CearÃ, northeast of Brazil, from July 2004 to December 2005. The Thirty who first came were selected to have R, I and P serum concentration dosed. Twenty nine healthy controls were select among health professionals, their relatives and neighboring. To access intestinal permeability lactulose/manitol (L/M) test was performed by HPLC in urine samples collected during five hours. To access bioavailability two blood samples were collect at 2 and 6 hours after drug ingestion. Demographic information was recorded from all volunteers. Mean age was 39.2  15 years in cases and 34  11.0 in controls (p=0.61); 71.4% cases and 65.5% controls were men (p=0.57). Lactulose urinary excretion was significantly lower in TB patients (median 0.1721%; range 0.0-5.0139) than in controls (median 0.4301%; range 0.2125-2.064) (p=0.0194); median manitol excretion in cases was 17.9910% (1.9567-71.5446) and in controls, 24.39894% (range 1.3883-63.0539) (p=0.147) and the lactulose/manitol ratio was of 0.0095 (range 0.0-0.0759) in cases and 0.0153 (0.0-0.0136) in controls (p=0.0698). Maximum means seric rifampin concentration in cases was1.46  0.72 Âg/ml and in controls, 3.07  6.69Âg/ml (p<0.001); maximum means seric isoniazid concentration was 2.62  1.53 Âg/ml in cases and 0.76  1.98 Âg/ml in controls (p=0.057); maximum means seric pirazinamide concentratio was 44.10  10.40 Âg/ml in cases and 12.02  36.32 Âg/ml in controls (p=0.007). Four cases (13.3%) had all tested drugs serum concentrations under expected normal; 21/30 (70.0%) had expected normal concentrations only for one drug (pyrazinamide) and 5/30 (16.7%) for 2 drugs only (pirazinamide and isoniazid). None of the cases had expected concentration levels for all 3 drugs, simultaneously. In conclusion, there was lesion in the functional intestinal barrier and malabsorption for rifampin and isoniazid in active pulmonary TB patients suggesting it is necessary a deeper evaluation of measures to reduce malabsorption, since only these drugs are used during last the four months of treatment

This study firstly reports the development and characterization of PVP-AgNPs, PANSA and PVPAgNPs/ PANSA nanocomposite on gold. AFM and TEM analyses revealed highly electroactive nanocomposites whose morphogy and properties were essential for the immobilization of CYP2E1. Secondly, the development and characterization of Au/PVPAgNPs/ PANSA/CYP2E1, Au/PVP-AgNPs/PANSA/SA-CYP2E1 and Au/PVPAgNPs/ PANSA/EG-CYP2E1 nanobiosensors are reported. AFM studies displayed globular morphologies with large roughness for the enzyme modified electrodes as opposed to those electrodes without enzymes. Finally, the biotransformation of standard solutions of TB drugs (isoniazid, ethambutol, pyrazinamide and rifampicin) in pH 7.4, 0.1 M phosphate buffer solution is reported. The biotransformations of the TB drugs were successfully studied using cyclic voltammetry (CV), square wave voltammetry (SWV), differential voltammetry (DPV) and steady state amperometry under aerobic conditions. Very good detection limits were obtained for the standard solutions of TB drugs and were found to be in the micromolar range. The detection limit values for the individual TB drugs were 0.55 μM (isoniazid), 0.7 μM (ethambutol), 0.054 μM (pyrazinamide) and 0.05 μM (rifampicin). The detection limit results showed that the nanobiosensors were more sensitive and suitable for the determination of the respective drugs in plasma and serum.

This study firstly reports the development and characterization of PVP-AgNPs, PANSA and PVPAgNPs/ PANSA nanocomposite on gold. AFM and TEM analyses revealed highly electroactive nanocomposites whose morphogy and properties were essential for the immobilization of CYP2E1. Secondly, the development and characterization of Au/PVPAgNPs/ PANSA/CYP2E1, Au/PVP-AgNPs/PANSA/SA-CYP2E1 and Au/PVPAgNPs/ PANSA/EG-CYP2E1 nanobiosensors are reported. AFM studies displayed globular morphologies with large roughness for the enzyme modified electrodes as opposed to those electrodes without enzymes. Finally, the biotransformation of standard solutions of TB drugs (isoniazid, ethambutol, pyrazinamide and rifampicin) in pH 7.4, 0.1 M phosphate buffer solution is reported. The biotransformations of the TB drugs were successfully studied using cyclic voltammetry (CV), square wave voltammetry (SWV), differential voltammetry (DPV) and steady state amperometry under aerobic conditions. Very good detection limits were obtained for the standard solutions of TB drugs and were found to be in the micromolar range. The detection limit values for the individual TB drugs were 0.55 μM (isoniazid), 0.7 μM (ethambutol), 0.054 μM (pyrazinamide) and 0.05 μM (rifampicin). The detection limit results showed that the nanobiosensors were more sensitive and suitable for the determination of the respective drugs in plasma and serum.

Philosophiae Doctor - PhD / This study firstly reports the development and characterization of PVP-AgNPs, PANSA and PVPAgNPs/ PANSA nanocomposite on gold. AFM and TEM analyses revealed highly electroactive nanocomposites whose morphogy and properties were essential for the immobilization of CYP2E1. Secondly, the development and characterization of Au/PVPAgNPs/ PANSA/CYP2E1, Au/PVP-AgNPs/PANSA/SA-CYP2E1 and Au/PVPAgNPs/ PANSA/EG-CYP2E1 nanobiosensors are reported. AFM studies displayed globular morphologies with large roughness for the enzyme modified electrodes as opposed to those electrodes without enzymes. Finally, the biotransformation of standard solutions of TB drugs (isoniazid, ethambutol, pyrazinamide and rifampicin) in pH 7.4, 0.1 M phosphate buffer solution is reported. The biotransformations of the TB drugs were successfully studied using cyclic voltammetry (CV), square wave voltammetry (SWV), differential voltammetry (DPV) and steady state amperometry under aerobic conditions. Very good detection limits were obtained for the standard solutions of TB drugs and were found to be in the micromolar range. The detection limit values for the individual TB drugs were 0.55 μM (isoniazid), 0.7 μM (ethambutol), 0.054 μM (pyrazinamide) and 0.05 μM (rifampicin). The detection limit results showed that the nanobiosensors were more sensitive and suitable for the determination of the respective drugs in plasma and serum. / South Africa

Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-05-13T18:09:13Z No. of bitstreams: 1 fabiodesouzafernandes.pdf: 24065820 bytes, checksum: da2a7ed64d78294c49d6e226495eda40 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-06-27T21:20:25Z (GMT) No. of bitstreams: 1 fabiodesouzafernandes.pdf: 24065820 bytes, checksum: da2a7ed64d78294c49d6e226495eda40 (MD5) / Made available in DSpace on 2016-06-27T21:20:25Z (GMT). No. of bitstreams: 1 fabiodesouzafernandes.pdf: 24065820 bytes, checksum: da2a7ed64d78294c49d6e226495eda40 (MD5) Previous issue date: 2016-02-29 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Na Química Medicinal, dentre as várias técnicas de planejamento racional para a síntese de novas moléculas bioativas, a hibridação é umas das mais usadas. A união de dois ou mais compostos biologicamente ativos, no intuito de se criar um novo fármaco que conserve as propriedades biológicas das moléculas base e/ou crie uma nova atividade biológica para o composto híbrido, faz desta técnica um caminho rápido, eficaz e oportuno no descobrimento de novos candidatos a fármacos. Sinteticamente, dentre os vários métodos de junção de duas ou mais moléculas a Reação de Mannich está entre as metodologias aplicadas para isto. Este primeiro capítulo trata da síntese N-bases de Mannich derivadas da pirazinamida, que resultou na obtenção de dezenove compostos inéditos. Em um primeiro momento, foi realizada a tentativa de síntese de N-bases de Mannich derivadas da pirazinamida, utilizando três diferentes aminoálcoois Nalquilados. O desenvolvimento da metodologia mostrou que a reação era dependente do pH, tendo este que ser básico. No entanto, a baixa conversão da reação, juntamente com a difícil purificação do produto, uma vez que o mesmo apresentava o mesmo fator de retenção da pirazinamida e alta instabilidade impedindo qualquer funcionalização in situ inviabilizou qualquer possibilidade de continuação desta síntese. Desta forma, foi feita a substituição dos aminoálcoois N-aquilados por piperazinas N-substituídas, contendo cadeias lipofílicas, uma porção D-galactose, amidas lipofílicas, aminoalcoóis aromáticos e heteroaromáticos, sendo sintetizados 5 derivados inéditos da piperazina. A partir da reação de Mannich das piperazinas Nsubstituídas com a pirazinamida foram obtidas quatorze N-Bases de Mannich inéditas. Tanto as piperazinas N-substituídas como as N-bases de Mannich sintetizadas, foram submetidas à avaliação de suas atividades antibacterianas, antibiofilme e antituberculose, sendo que os resultados para atividade antibiofilme apresentaram bastante promissores. As lactamas são amidas cíclicas presentes em vários produtos naturais e em compostos sintéticos com atividade biológica. Devido a esta importância dentro da química medicinal, várias são as estratégias desenvolvidas para a síntese destes anéis heterocíclicos, com o objetivo de se conseguir uma síntese rápida, eficiente, estereoseletiva e utilizando a química verde para destes compostos. Dentre as várias metodologias sintéticas que permitem a obtenção estereoseletiva de Lactamas está à reação de Mannich-Acilação, inicialmente desenvolvida por Castagnoli em 1969 e aperfeiçoada recentemente pelo grupo de pesquisa do Professor Dr. Jared Shaw. Sendo assim, o segundo capítulo aborda desenvolvimento de uma metodologia a paritr da reação de Mannich-Acilação para a síntese de δ-lactamas como suporte para um estudo computacional sobre o mecanismo e estereoseletividade da reação. Para isso, a síntese do anidrido altamente reativo α-ciano glutárico, bem como, dos anidridos α- ciano glutárico substituídos com grupos metila ou fenila nas posições β ou γ foram realizadas. O escopo e as limitações da reação imina-anidrido com uma ampla variedade de iminas também foi investigada. As reações contendo aminas substituídas por grupos alquil volumosos e aromáticos foram mais diastereoseletivas. Já as reações envolvendo o uso dos anidridos α-ciano glutáricos não substituídos e contendo os grupos metila e fenila na posição β mostraram-se altamente diastereoseletivas, enquanto que para as reações envolvendo o anidrido α-ciano glutárico substituídos na posição γ houve uma perda da seletividade. Os estudos computacionais mostraram que a origem do estereocontrole da reação se dá na etapa de acilação, onde efeitos estéreos e trans anulares para o estado de transição na forma eclipsada justificam a formação do produto majoritário tendo uma relação syn entre os grupos carboxilato e fenil. Neste capítulo também foram exploradas algumas aplicações, no que diz respeito à utilização das δ-lactamas como possíveis inibidores da proteína FtsZ, em um estudo onde as δ-lactamas serviram como substratos na reação clássica de descarboxilação alilativa e na síntese dos produtos naturais Gelsidilam e Gelgamina B tendo a reação de Mannich-Acilação desenvolvida neste capítulo como uma das etapas chave. / In the Medicinal Chemistry, there are many techniques for development of new bioactive molecules. One way to do a rational planning of new drugs is using the molecular hybridization technique. In the molecular hybridization, the combination of two or more biologically active compounds in order to create a new pro-drug that preserves the biological properties or improve efficacy when compared to the parent drugs, makes this technique a quick way, effective and timely in the discovery of new drug candidates. Mannich reaction can be applicable to combine two or more fragments of drugs with biological activity. The first chapter will show the synthesis of new N-Mannich bases derived from pyrazinamide, which resulted in the synthesis of nineteen novel compounds. First of all, we tried to make the synthesis of new N-Mannich bases from of reaction between pyrazinamide, formaldehyde and N-alkylated amino alcohols. The development of this methodology showed that the reaction was pH dependent and it had to be basic. So we tried to control the pH of reaction using some bases, but the lower conversion of reaction together with the difficult purification of the product and the high instability led to an unsuccessful planning to get the products desired. Because of this, was made replacing of N-alkylated amino alcohols by N-substituted piperazines having lipophilic chain, D-galactose moiety, lipophilic amides, aromatic and heteroaromatic amino alcohols. With these new components we ran the Mannich reactions and was possible to synthesize fourteen novel N-Mannich bases. The N-substituted piperazines and N-Mannich bases synthesized were evaluated by antibacterial, antibiofilm and antituberculosis activities. The results of antibiofilm and antibacterial activity for some hybrids were very promising. In the second chapter, was developed a methodology to the synthesis of δ- lactams from cycloaddition between imines and cyano glutaric anhydrides like a support to mechanism and diastereoselectivity study of reaction. For this was made the synthesis of β, γ-substituted and unsubstituted cyano anhydrides. The scope of reaction was studied using differents amines and aldehydes. The reactions that were ran with bulky alkyl-substituted amines or aromatic amines were more selective than methylamine. The reactions involving the use of α-cyano-glutaric anhydrides unsubstituted or containing methyl groups and phenyl in the β position showed high diastereoselectivity. Reactions using the γ substituted anhydrides had a poor diastereoselectivity. Computational studies showed that acylation step is a determinant factor to diastereoselectivity, where the eclipsed transition state to anti product has steric and trans anular effects. Some applications of -lactams were made in other studies involving Tethering technique, decarboxylative allylation and total synthesis of Gelsedilam and Gelegamine B.

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No. of bitstreams: 1 adrianaferreirafaria.pdf: 20849316 bytes, checksum: 511836cab839ebb39105bd682c987241 (MD5) Previous issue date: 2010-08-13 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O objetivo principal da tese em questão foi o desenvolvimento, otimização e validação de metodologias analíticas para análise de formulações farmacêuticas utilizadas no tratamento da tuberculose, as quais contêm como princípio ativo etambutol (ETB), isoniazida (ISO), rifampicina (RIF) e pirazinamida (PIR), isolados ou em associação. Inicialmente, duas metodologias analíticas para análise de ETB em comprimidos foram desenvolvidas: a primeira permitiu a análise simultânea de ETB e da impureza 2-amino-1-butanol por eletroforese capilar de zona (CZE) utilizando um eletrólito de corrida contendo 60,0 mmol L-1 de tampão ácido acético/acetato de sódio e 5,0 mmol L-1 de Cu2+, polaridade normal (+25 kV) e detecção UV em 262 nm. A segunda foi realizada por espectrofotometria com diluição dos padrões e amostras em 5,0 mmol L-1 de tampão ácido acético/acetato de sódio e 0,5 mmol L-1 de Cu2+ e detecção UV em 262 nm. A formação do complexo CuETB foi necessária em ambas as metodologias desenvolvidas, uma vez que ETB apresenta baixa absortividade molar. Algumas figuras de mérito, tais como, linearidade, repetitividade, exatidão, limite de detecção e limite de quantificação foram avaliadas para as duas metodologias. Os dois métodos foram comparados e não apresentaram diferenças significativas no intervalo de 95% usando o teste t não pareado com variância agrupada. Em um segundo momento, um novo método para análise simultânea de dose fixa combinada (DFC), contendo ETB, ISO, RIF e PIR por CZE com detecção UV foi desenvolvido, otimizado e validado. A condição experimental consistiu em um eletrólito contendo 50,0 mmol L-1 de tampão ácido acético/acetato de sódio e 12,5 mmol L-1 de Cu2+ e polaridade normal (+22 kV). Essa metodologia foi aplicada com sucesso na análise de 2-DFC (ISO e PIR) em comprimidos e 4-DFC (ETB, ISO, RIF e PIR) em sache. Finalmente, uma metodologia por CZE utilizando detecção UV para a análise simultânea de ISO, suas impurezas e produtos de degradação, como o ácido isonicotínico, isonicotinato de etila, 4-cianopiridina e isonicotinamida foi otimizada. A condição experimental consistiu em um eletrólito contendo 50,0 mmol L-1 de tampão ácido acético/acetato de sódio e 12,5 mmol L-1 de Cu2+, polaridade normal (+20 kV) e detecção UV em 270 nm. É importante ressaltar que planejamentos de experimentos, tais como, planejamento fatorial completo 32 e Box-Behnken 33 foram utilizados como ferramenta auxiliar no desenvolvimento e otimização das metodologias analíticas (CZE e espectofotométrica) e na avaliação da robustez para a análise de 4-DFC. / The main focus of this thesis was the development, optimization and validation of analytical methodologies for analysis of pharmaceutical preparations used for tuberculosis treatment, which contain as active ingredients, ethambutol (ETB), isoniazid (ISO), rifampicin (RIF) and pyrazinamide (PYR) isolated or in association. Two analytical methodologies for ETB analysis in tablets were developed initially: the first one took into account the capillary zone electrophoresis (CZE) approach for simultaneous analysis of ETB and its impurity 2-amino-1-butanol using an electrolyte consisting of 60.0 mmol L-1 of acetic acid/acetate buffer, 5.0 mmol L-1 of Cu2+, under normal polarity (+25 kV) and UV detection at 262 nm. The second one was performed by spectrophotometry using 5.0 mmol L-1 of acetic acid/acetate buffer and 0.5 mmol L-1 of Cu2+ for standards and sample dilutions under UV detection at 262 nm. The CuETB complex was necessary for the two methodologies, since ETB presents low molar absorptivity. Some merit figures such as linearity, repeatability, accuracy, limit of detection and limit of quantification were evaluated for both methodologies. These methodologies were compared and they did not present significant differences at the 95% confidence level using the parametric independent sample t test. In a second phase a novel method for the simultaneous analysis of fixed dose combination (FDC) containing ETB, ISO, RIF and PYR by CZE under UV detection was developed, optimized and validated. The experimental condition optimized consisted of an electrolyte containing 50.0 mmol L-1 of acetic acid/acetate buffer and 12.5 mmol L-1 of Cu2+ under normal polarity (+22 kV). This methodology was successfully applied to the analysis of 2-FDC (ISO and PYR) in tablet samples and 4-FDC (ETB, ISO, RIF and PYR) in a sachet sample. Finally, a CZE methodology using UV detection for simultaneous analysis of ISO, its impurities and degradation products such as isonicotinic acid, ethyl isonicotinate, 4-cyanopyridine and isonicotinamide was ptimized. The optimized experimental condition consisted of an electrolyte containing 50.0 mmol L-1 of acetic acid/acetate buffer and 12.5 mmol L-1 of Cu2+ under normal polarity (+20 kV) and UV detection at 270 nm. It is important to stress that design of experiment such as 32 full factorial design and 33 Box-Behnken design were used as auxiliary tools in the development and optimization of the analytical methodologies (CZE and spectrophotemetric) and robustness evaluation for 4-FDC analysis.

Die Mechanochemie findet zunehmend Verwendung für die Synthese neuer Verbindungen. Dennoch sind die beim Mahlen stattfindenden Prozesse weitestgehend unverstanden. Dahingehend wurde in dieser Arbeit eine Dreifachkopplung aus in situ Synchrotron-Röntgenbeugung, Raman-Spektroskopie und Thermographie entwickelt, um mechanochemische Reaktionen unter realistischen Bedingungen in Echtzeit zu verfolgen. Dadurch konnten tiefgreifende Einblicke in den Reaktionsverlauf und Temperaturverlauf beim Mahlen erhalten und neue metastabile Verbindungen isoliert werden. Für die Bildung pharmazeutischer Cokristalle diente Pyrazinamid als Modellsystem. Es konnten neue binäre und ternäre Verbindungen synthetisiert, detailliert charakterisiert und deren Kristallstruktur aufgeklärt werden. Die Abhängigkeit der Stabilität polymorpher Cokristalle von der Temperatur und den Synthesebedingungen konnte gezeigt werden. In Konkurrenzreaktionen konnten Trends hinsichtlich der bevorzugten Bildung eine bestimmten Cokristalls beobachtet werden. Mittels in situ Untersuchungen wurde der Einfluss zentraler Reaktionsparameter, wie die Mahlfrequenz, der Kugeldurchmesser, der eingesetzte Ausgangsstoff und die Zugabe von Lösungsmittel, auf die Induktions- und Reaktionszeit der Reaktion ermittelt. Basierend auf den gewonnenen Erkenntnissen konnte ein Diffusionsmechanismus für die mechanochemische Cokristallbildung abgeleitet werden. / Mechanochemistry is increasingly applied for the synthesis of new compounds. Still, the processes taking place during milling are far from being understood. In this thesis, a triple coupling of in situ synchrotron X-ray diffraction, Raman spectroscopy and thermography has been developed to follow mechanochemical reactions under realistic conditions in real time. This allowed deep insights into the reaction and temperature progression during milling and the isolation of new metastable compounds. For the formation of pharmaceutical cocrystals pyrazinamide served as a model system. New binary and ternary compounds were synthesized, characterized in detail and their crystal structure solved. The dependence of the stability of polymorphic cocrystals on temperature and synthesis conditions could be shown. In competitive reactions, trends regarding the preferred formation of a certain cocrystal have been observed. The influence of important reaction parameters, such as the milling frequency, the ball diameter, the starting material used and the addition of solvent, on the induction and reaction time of the reaction was determined by means of in situ investigations. Based on the gained knowledge, a diffusion mechanism for the mechanochemical cocrystal formation could be derived.

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No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Bruno_Regis_Lyrio_Ferraz.PDF: 3015137 bytes, checksum: d479e9335e2f200939b52075f9b84382 (MD5) Previous issue date: 2016 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Funda??o de Amparo a Pesquisa do Estado de Minas Gerais (FAPEMIG) / RESUMO Etionamida e pirazinamida s?o antibi?ticos ?teis no tratamento da tuberculose multirresistente. O presente trabalho descreve o desenvolvimento e valida??o de metodologias eletroanal?ticas para determina??o de etionamida e pirazinamida em formula??es farmac?uticas e em urina humana empregando um eletrodo de diamante dopado com boro e um eletrodo de carbono v?treo modificado comum filme de poli glicina. Durante o desenvolvimento de ambas as metodologias, a voltametria c?clica foi empregada para verificar a influ?ncia do pH, da velocidade de varredura e do eletr?lito suporte no comportamento eletroqu?mico de ambos os analitos, bem como foram calculados os n?meros de pr?tons e el?trons envolvidos em cada uma das rea??es eletroqu?micas. A voltametria de onda quadrada com os par?metros otimizados foi utilizada para construir curvas anal?ticas para a ETO e PZA. Para a ETO foi obtido um intervalo linear de 1,0 a 80,0 ?mol L?1, com LOD e LOQ iguais a 0,294 e 0,980 ?mol L?1, respectivamente. Para a PZA foi obtido um intervalo linear de 0,47 a 6,16 ?mol L?1, com LOD e LOQ iguais a 0,035 e 0,12 ?mol L?1, respectivamente. A precis?o foi avaliada pelo registro de voltamogramas no mesmo dia e em dias diferentes, obtendo-se desvios padr?es relativos, inferiores a 5,0% em ambos os m?todos. Os resultados dos estudos de interferentes mostraram que nenhuma das subst?ncias testadas interferiu de maneira significativa na determina??o de ambos os f?rmacos. Os m?todos desenvolvidos foram comparados estatisticamente com os protocolos oficiais da farmacopeia atrav?s do teste-t e do teste-F, e os resultados mostraram que os valores de t e F calculados foram menores do que os valores de t e F cr?ticos, indicando que n?o houve diferen?a estat?stica entre as m?dias. A exatid?o de ambos os m?todos foi avaliada tamb?m por estudos de adi??o e recupera??o, obtendo-se como resultados percentuais de recupera??o pr?ximos a 100% para ambos os m?todos. A valida??o das metodologias desenvolvidas foi realizada pela avalia??o dos par?metros anal?ticos como sensibilidade, seletividade, limite de detec??o, limite de quantifica??o, faixa linear, exatid?o e precis?o e os resultados obtidos foram satisfat?rios. Portanto, os m?todos desenvolvidos podem ser aplicados com sucesso na determina??o dos f?rmacos ETO e PZA em medicamentos e urina humana. / Disserta??o (Mestrado) ? Programa de P?s-gradua??o em Ci?ncias Farmac?uticas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2016. / ABSTRACT Ethionamide and pyrazinamide antibiotics are useful in the treatment of multidrugresistant tuberculosis. This work describes the development and validation of electroanalytical methodologies for determination of ethionamide and pyrazinamide in pharmaceutical formulation and human urine using boron-doped diamond electrode and poly glycine modified glassy carbon electrode, respectively. During the development of both methodologies, cyclic voltammetry was used to investigate the influence of pH, scan rate and the supporting electrolyte on the electrochemical behavior of both analytes, as well as the numbers of protons and electrons involved in each of the electrochemical reactions were calculated. Square wave voltammetry with optimized parameters were used to construct standard curves for ETO and PZA. For ETO a linear range from 1.0 to 80.0 ?mol L?1 was obtained with LOD and LOQ equal to 0.294 and 0.980 ?mol L?1, respectively. For PZA a linear range from 0.47 to 6.16 ?mol L?1was obtained with LOD and LOQ equal to 0.035 and 0.12 ?mol L?1, respectively. The precision was evaluated by voltammograms record on the same day and on different days, obtaining relative standard deviation less than 5.0% in both methods. The results of interfering studies showed that none of the tested substance interferes significantly in the determination of both drugs. The developed methods were statistically compared with the pharmacopoeia official protocols through the t-test and F-test, and the results showed that the calculated t and F values were lower than the critical t and F values indicating that there was no statistical difference between the averages. The accuracy of both methods was also evaluated by addition and recovery studies, obtaining results as percentage recovery close to 100% for both methods. The validation of the developed methodologies was carried out by the evaluation of analytical parameters such as sensitivity, selectivity, detection limit, quantification limit, linear range, accuracy and precision and the obtained results were satisfactory. Therefore, the developed methods can be applied successfully in the determination of ETO and PZA drugs in pharmaceuticals and human urine.