Dissertations / Theses on the topic 'Acetazolamide'

A fotoestabilidade é uma propriedade das moléculas que, quando utilizada como parâmetro farmacêutico, descreve como um fármaco responde à exposição à luz (solar ou artificial). No presente trabalho, foi avaliada a fotoestabilidade dos fármacos loratadina (LORA) e acetazolamida (ACZ) e de complexos LORA-ciclodextrinas. O estudo de fotoestabilidade de LORA (Capítulo 2) indicou que o fármaco é estável quando no estado sólido, porém, ocorre surgimento de coloração intensa. Por outro lado, quando em solução, observou-se degradação do fármaco, com surgimento de vários fotoprodutos denominados F1 a F15, dentre os quais foi possível identificar cinco compostos: F4 (C13H10N), F10 (C14H10CIN), F8 (C20H18CIN2O), F9 (C19H18CIN2) e F14 (C17H14CIN). A validação do método analítico CLAE, utilizado para quantificação de LORA em especialidades farmacêuticas (comprimidos e xaropes) é descrita no Capítulo 3. Na avaliação da fotodegradação forçada de formulações líquidas contendo LORA, foram degradados até 50% do fármaco. As formulações sólidas apresentaram-se fotoestáveis, observando-se perda de menos de 5% do fármaco. Não foram encontrados produtos de fotodegradação nas formulações, quando analisadas tal qual, obtidas do mercado. Dessa forma, as embalagens primárias garantiram sua estabilidade. A complexação de LORA com ciclodextrinas (Capítulo 4) mostrou-se um recurso bastante interessante para melhorar a fotoestabilidade do fármaco, uma vez que, após 12 horas de irradiação luminosa, é possível recuperar até 99% deste, quando na forma de complexo com γ-CD na proporção 1:1. Finalmente, o Capítulo 5 traz o método CLAE desenvolvido e validado para avaliação da acetazolamida (ACZ), o qual mostrou-se adequado para a quantificação do fármaco, obtendo-se ótima linearidade, precisão, exatidão e seletividade. Segundo as condições do guia Q1B, a ACZ se manteve estável quando submetida à radiação luminosa utilizando meios aquosos e no estado sólido. No entanto, a fotoestabilidade da ACZ foi afetada na presença de metanol, sendo possível quantificar três impurezas.
Photostability is a property of molecules that, when used as a pharmaceutical parameter, can describe how a drug responds to exposure to light (either solar or artificial). In this study, the photostability of the drugs loratadine (LORA) and acetazolamide (ACZ), as well as LORA-cyclodextrin complexes, was evaluated. A study of the photostability of LORA (Chapter 2) indicated that the drug is stable in its solid form, however intense coloring does occur. On the other hand, when in solution form, degradation of the drug was observed, with the appearance of several photoproducts that we labled F1 to F15, among which it was possible to identify five compounds: F4 (C13H10N), F10 (C14H10CIN), F8 (C20H18CIN2O), F9 (C19H18CIN2) and F14 (C17H14CIN). The validation of the analytical method by HPLC, used for the quantification of LORA in pharmaceutical products (tablets and syrups) is detailed in Chapter 3. In the evaluation of forced photodegradation of liquid formulations containing LORA, up to 50% of the drug was degraded. The solid formulations proved to be photostable, with a loss of less than 5% of the drug. No photodegradation products were found in the formulations when they were analyzed \"as is\" (the way they were obtained from the commercial market). Accordingly, their primary packaging protected their stability. The complexation of LORA with cyclodextrins (Chapter 4) proved to be an effective resource for improving the photostability of the drug, since, after 12 hours of luminous radiation, it was possible to recover up to 99% of the drug, when in the complex form with γ-CD, in the proportion 1:1. Finally, Chapter 5 describes the HPLC method developed and validated for the evaluation of acetazolamide (ACZ), which proved to be adequate for the quantification of the drug, with the attainment of optimal linearity, precision, exactness and selectivity. According to the conditions of the Q1B guideline, ACZ was stable when subjected to luminous radiation using aqueous means and in its solid state. However, the photostability of ACZ was affected by the presence of methanol, and we were able to quantify three impurities.

Exposing Amyloid Beta 1-42 to neurons causes cell death. When carbonic anhydrase inhibitors (e.g. methazolamide or acetazolamide) are introduced along with 1-42 in a similar experiment, cell apoptosis is disrupted. However, when non-CA inhibitors are tested, (e.g. the indole derivative melatonin), the same disruption occurs. Are these carbonic anhydrase inhibitors acting on the same or a different pathway? One way to study the molecular mechanisms of these small molecule inhibitors is to modify their chemical structure. In this sense, when acetazolamide is methylated, apoptosis is resumed (Fossati et al., 2016). Finding a way to create N-methyl acetazolamide and N-methyl methazolamide through methylation procedures will lead to a better understanding of the pathways involved in neuronal apoptosis triggered by the Abeta peptide.

Context: Acetazolmaide (AZ) is used in the prophylactic treatment of altitude illnesses. AZ is also known to attenuate HPV and increase alveolar ventilation. Methazolamide (MZ) is an analog to AZ and its effects on ventilation and HPV are unknown. Objective. To determine if MZ can improve oxygenation and attenuate HPV to a similar extent as AZ in healthy humans exposed to poikilocapnic hypoxia. Design, Setting, Participants and Interventions: A randomized, placebo controlled, double-blinded trial was performed in healthy participants at the Cardiopulmonary Lab for Experimental and Applied Physiology. Prior to each of the three experimentation days, participants were administered one of three treatments (AZ, MZ, & placebo) at random for two days. Each treatment was separated by a 10-day washout period to avoid contamination from previous trials. During each trial, participants were exposed to poikilocapnic hypoxia (FIO2 ≈ 0.12) for 60 minutes. Primary Outcome Measures: Partial pressure of alveolar O₂ (PAO₂) represented oxygenation while pulmonary artery systolic pressure (PASP) and total pulmonary resistance (TPR) were chosen to represent the HPV response. Results: All participants (n = 11) completed all three trials. Change in Q̇ from baseline to hypoxia was not different between treatments. Change in PASP was significantly lower with the AZ (8.0 ± 0.7 mmHg) and MZ (9.0 ± 0.9 mmHg) treatments compared to placebo (PASP: 14.1 ± 1.3 mmHgP < 0.05). Change in PAO₂ was also decreased with both drug treatments (AZ: 54.8 ± 1.3 mmHg; MZ: 53.9 ± 1.3 mmHg) compared to placebo (48.5 ± 1.6 mmHg; P < 0.05). Conclusion: MZ attenuated HPV to the same degree as AZ. MZ also resulted in a similar improvement in PAO₂ as AZ during hypoxia compared to placebo. Trial Registration: This study was registered with the U.S. National Institutes of Health: NCT02760121 Funding: This project was funded by the Canadian Foundation for Innovation and by the Natural Science and Engineering Council of Canada.
Graduate Studies, College of (Okanagan)
Graduate

This study was designed to determine the effect of caffeine on the teratogenicity of acetazolamide in susceptible and resistant strains of inbred mice. The highly susceptible C57BL/6J and more resistant SWV strain were used. Pregnant C57BL/6J and SWV mice were treated with caffeine, low or high dose acetazolamide, or a combination of both agents during the sensitive period of development. Untreated and vehicle-treated groups served as controls. Individual fetuses were examined for gross morphological abnormalities and skeletal variations.Findings1. A highly significant (P<.001) increase in fetal malformations, especially right forelimb ectrodactyly, was evident in C57BL/6J litters exposed on day 9 of gestation to both agents when contrasted with those exposed to either agent alone. Both frequency and severity of ectrodactyly was potentiated by caffeine.2. The SWV strain was resistant to the interaction of caffeine and acetazolamide when treated on day 9 of gestation. However, a highly significant (P<.001) increase in the rate of fetal malformation was found in litters whose dams were treated with high dose acetazolamide and caffeine on day 8 of gestation. The most common malformations observed were exencephaly and umbilical hernia.3. No differences in maternal mortality, fetal weight, litter size, or embryo mortality could be attributed to treatment in either strain.4. Skeletal examination of the number of ossified cervical and caudal vertebral centra revealed a reduction in ossification among C57BL/6J litters exposed to high dose acetazolamide or acetazolamide plus caffeine. These same centers of ossification were mildly affected by treatment in the SWV strain. In both strains the first cervical vertebrae (Cl) appeared to provide the most sensitive index of teratogenic exposure.ConclusionsThis study provides evidence that a subteratogenic dose of caffeine can potentiate the teratogenic effect of acetazolamide in both C57BL/6J and SWV mice. However, strain associated, and therefore genetically based, differences in sensitivity were confirmed. Skeletal examinations provided evidence that treatment with both agents delayed fetal development in the more susceptible C57BL/6J strain, while reduction of ossification was less evident in the SWV strain. Thus, this parameter also reflects the greater resistance of the SWV strain to the interaction of acetazolamide and caffeine. Finally, these experiments support the idea that chemical interactions may, in part, be responsible for many birth defects of unknown etiology--a claim worthy of further investigation.
Department of Biology

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior
As doenÃas periodontais sÃo as principais causas de perda de dentes em adultos. O fator etiolÃgico preponderante para desencadear a periodontite à o acumulo do biofilme dental com predominÃncia de bactÃrias anaerÃbicas gram negativas. Apesar de ser de origem bacteriana a resposta inflamatÃria induzida a partir de fatores do hospedeiro pode influenciar na progressÃo e nas caracterÃsticas clÃnicas da doenÃa periodontal. A perda Ãssea alveolar à tÃpica na evoluÃÃo desta patologia e dependente da atividade osteoclÃstica. Por sua vez, um dos mecanismos de reabsorÃÃo Ãssea por estas cÃlulas à atravÃs da produÃÃo de prÃtons liberados nos vacÃolos de reabsorÃÃo pela atividade da anidrase carbÃnica. Tem sido relatado que inibidores desta enzima, como acetazolamida, tem aÃÃo supressora sobre osteoclastos. O objetivo deste estudo foi analisar o efeito da acetazolamida (ACTZ) na perda Ãssea induzida no modelo de periodontite experimental em ratos, bem como a dosagem de biomarcadores sÃricos do processo inflamatÃrio periodontal e anÃlise histolÃgica do periodonto. AnÃlises de seguranÃa tambÃm foram realizadas atravÃs de hemograma completo, dosagens de TGP, TGO, gama-GT e histopatologia de ÃrgÃos como: fÃgado, rins, baÃo, coraÃÃo e pulmÃo. Para avaliaÃÃo da perda Ãssea alveolar 50 ratos receberam ligaduras de nylon na regiÃo do segundo molar superior esquerdo. Os animais foram divididos em seis grupos: Grupo veÃculo (propilenoglicol) (n=9), grupo controle positivo (alendronato 0,08 mg/Kg) (n=7), grupo controle negativo (salina) (n=9) e grupos que receberam acetazolamida via I.P. nas doses (8,3, 25 e 75 mg/Kg [grupos ACTZ8,3, ACTZ25 e ACTZ75, respectivamente] n= 7, 9 e 9). ApÃs 11 dias, os ratos foram mortos. A perda Ãssea alveolar foi avaliada macroscopicamente atravÃs da Ãrea de exposiÃÃo de raiz. Em outros quatro grupos, ACTZ25 (n=5) e veÃculo (n=5), o periodonto foi analisado histologicamente apÃs 11 e 17 dias de ligadura por coloraÃÃo em HE. Biomarcadores sÃricos foram dosados (IL-1, IL-4, fractalkine, CINC-2, CINC-3, LIX, GM-CSF, βNGF, VEGF e CNTF) atravÃs da tÃcnica de microarray antes e depois da ligadura em trÃs grupos; ACTZ25, Veiculo e Sham (cirurgia simulada). No grupo de ACTZ25 foi realizada a anÃlise de seguranÃa (alpha=5%). Macroscopicamente, o grupo alendronato (controle positivo) apresentou a menor perda Ãssea, seguido do grupo ACTZ75 e ACTZ25 (p<0.05) quando comparados aos controles veÃculo e salina que nÃo diferiram entre si (p>0.05). Histologicamente, o grupo ACTZ apresentou a menor reabsorÃÃo de osso e cemento (p<0,05) apÃs 17 dias de ligadura, nÃo houve diferenÃa entre o grupo ACTZ25 e o veÃculo quanto ao infiltrado inflamatÃrio (P>0,05) . As maiores concentraÃÃes de IL-4 e CNTF foram observadas no grupo ACTZ25 (p< 0,05) quando comparado ao grupo veÃculo. As anÃlises de seguranÃa demonstraram que acetazolamida na dosagem de 25 mg/Kg foi bem tolerada sem alteraÃÃes significativas do hemograma, nas enzimas hepÃticas e histopatologia dos ÃrgÃos pesquisados. ConcluÃmos que a acetazolamida pode proteger o periodonto da reabsorÃÃo Ãssea induzida por ligadura em ratos e pode estar associada a mediadores envolvidos com o reparo, como a IL-4 e CNTF, alÃm de ser bem tolerada.
Periodontal diseases are the main cause of tooth loss in adults. The preponderant etiologic factor for the onset of periodontitis is the accumulation of dental biofilm with predominance of anaerobic gram negative bacteria. In spite of being of bacterial origin, the inflammatory response induced by factors in the host may influence the progression and clinical characteristics of periodontal disease. Alveolar bone is typical in the development of this pathology and is dependent on osteoclastic activity. In turn, one of the mechanisms of bone resorption by these cells is through the production of protons released in the resorption vacuoles by carbonic anhydrase activity. It has been reported that carbonic anhydrase inhibitors, such as acetazolamide, have a suppressive action on osteoclasts. The aim of this study was to analyze the effect of acetazolamide (ACTZ) on bone loss induced in the experimental periodontitis model in rats, as well as the dosage of seric biomarkers in the periodontal inflammatory process and histologic analysis of the periodontium. Safety analyses were also performed by means of a complete hemogram, biochemical tests for TGP, TGO, gama-GT and histopathology of organs such as: the liver, kidneys, spleen, heart and lungs. To evaluate alveolar bone loss, 50 rats received nylon ligatures in the maxillary left second molar region. The animals were divided into six groups: Vehicle Group (propylenoglycol) (n=9), Positive Control Group (alendronate 0.08 V (n=7), Negative Control Group (saline) (n=6) and Groups that received acetazolamide I.P. in the following doses: (8.3, 25 and 75 mg/Kg [Groups ACTZ8,3, ACTZ25 and ACTZ75 respectively] n= 7, 9 and 9). After 11 days, the rats were sacrificed. Alveolar bone loss was macroscopically evaluated by means of the area of root exposure. In another four groups, ACTZ25 (n=5) and vehicle (n=5), the periodontium was histologically analyzed after 11 and 17 days of ligature, by HE staining. Seric biomarkers were dosed (IL-1, IL-4, fractalkine, CINC-2, CINC-3, LIX, GM-CSF, βNGF, VEGF and CNTF) by means of the microarray technique before and after ligature in three groups; ACTZ25, Vehicle and Sham (simulated surgery). In the ACTZ25 group, safety analysis was performed (alpha=5%). Macroscopically, the alendronate Group (positive control) presented the lowest bone loss, followed by Groups ACTZ75 and ACTZ25 (p<0.05) when compared with the vehicle and saline controls, which did not differ between them (p>0.05). Histologically, the ACTZ group presented the lowest bone and cement resorption (p<0.05) after 17 days of ligature, and there was no difference between the ACTZ25 and vehicle groups with regard to inflammatory infiltrate (P>0.05). The highest concentrations of IL-4 and CNTF were observed in Group ACTZ25 (p< 0.05) when compared with the vehicle group. The safety analyses demonstrated that acetazolamide at the dose of 25 mg/Kg was well tolerated without significant alterations in the hemogram, hepatic enzymes and histopathology of the researched organ. It was concluded that acetazolamide may protect the periodontium from bone resorption induced by ligature in rats, and may be associated with mediators involved in repair, such as IL-4 and CNTF, in addition to being well tolerated.

The study was designed to determine what type of potentiation,.if any, occurred between caffeine and acetazolamide. Caffeine (75 mg/kg) and/or acetazolamide (200, 1000, or 1500 mg/kg) were administered to pregnant C57BL/6J dams on day 9 of gestation. Fetuses were removed on the eighteenth day of gestation via cesarean section and examined for gross morphological malformations using a Bausch & Lomb SKV1070P dissecting microscope. Treatment with HD-ACZM and HD+CAFF resulted in a reduction of fetal weight. Maternal exposure to MD-ACZM and HD-ACZM caused a statistically significant (P < .001) and dose-dependent increase in the percent of C57BL/6J fetuses with ectrodactyly along with increased severity of the defects displayed (relative to controls). An increase in the number of ectrodactylous fetuses and the severity of defects was also observed in all groups administered caffeine and acetazolamide, reaching statistical significance in the MD+CAFF and HD+CAFF groups (P < .001). Because potentiation of the teratogenic effects of acetazolamide was exhibited only in the MD+CAFF vs. MD-ACZM groups, the type of potentiation occurring between caffeine and acetazolamide can not be determined.
Department of Physiology and Health Science

Acetazolamide (AZA), the only drug approved for treatment of hydrocephalus, is effective in only 25-30% of patients while its effect on fluid flow in the choroid plexus (CP) is unknown. The drug reversibly inhibits Aquaporin 4 (AQP4), the most highly expressed 'water pore' in the brain, and it is postulated that it reduces cerebrospinal fluid (CSF) production by modulating AQP1 (mostly found in the apical membrane of the CP). In this study, we sought to elucidate the effect of AZA on AQP1 and fluid flow in CP. Primary CP culture from p10 Sprague-Dawley rats and TRCSF-B cell line were grown on Transwell permeable supports, treated with 100micrometers] AZA or 100micrometers] Vinpocetine (previously shown to increase AQP1 levels), and tested by: a) Fluid assays using TRITC-labeled Dextran to assay direction and extent of fluid flow; b) Immunoblot, Immunocytochemistry (ICC), and RT-PCR for AQP1 expression. Immnoblots and ICC analyses showed that AQP1 protein levels decrease in a delayed manner (lowest at 12 hours) with AZA treatment. The reduction in AQP1 protein was transient and preceded by a reduction in mRNA levels (lowest at 6 hours). Transwell fluid assays indicate a shift in fluid flow at 2 hours, prior to the changes in AQP1 mRNA or protein. Alteration of fluid flow by AZA (in both primary culture and TR-CSFB) is similar to Vinpocetine's effect in primary culture. Together with drug-induced alterations in AQP1 levels, these data suggest independent mechanisms behind fluid flow and AQP1 expression.
ID: 030423005; System requirements: World Wide Web browser and PDF reader.; Mode of access: World Wide Web.; Thesis (M.S.)--University of Central Florida, 2010.; Includes bibliographical references (p. 48-52).
M.S.
Masters
Burnett School of Biomedical Sciences
Medicine
Biotechnology

Aquaporin (AQP) water channels are critical components of the urine concentrating mechanism in mammalian kidneys, and to-date, seven homologs of this channel have been found to play a role in renal physiology. In particular, Aquaporin-1 (AQP1) is located along the proximal tubule (PT), thin descending limb of the loop of Henle (tDLLH), and descending vasa recta (DVR). Also important in the kidneys are α- and β-intercalated cells (ICs), which regulate pH and are found in the distal tubule (DT) and collecting duct (CD). In the AQP1-knockout mouse model, CDs have been shown to demonstrate increased expression of α-ICs, suggesting compensatory increases in bicarbonate reabsorption and proton secretion. Two important pH modulating mechanisms in α-ICs are the carbonic anhydrase catalyzed interconversion of carbon dioxide and water to bicarbonate, and luminal and abluminal chloride/bicarbonate antiporter activity. The purpose of this study is to investigate differences in renal pH regulation between wild-type (WT) and AQP1-knockout (KO) mice. In particular, we aim to measure the r1 MRI relaxivities of Gd-DTPA, Gd-DOTA-4AmP, and Gd.3a at 7 Tesla in vitro, employ Dynamic Contrast-Enhanced Magnetic Resonance Imaging (DCE-MRI) in vivo in order to generate co-registered maps of renal functional parameters in wild-type and AQP1-knockout mice, and investigate the pharmacodynamics of acetazolamide and DIDS in wild-type and AQP1-knockout mice. We have compared wild-type and AQP1-knockout mice treated with saline, acetazolamide (a carbonic 9 anhydrase inhibitor), 4,4-diisothiocyano stilbene-2,2-disulfonic acid (DIDS, a Cl-/HCO3- exchange inhibitor), and untreated mice. We have imaged tissue pH in these mice by DCE-MRI using standard and pH-sensitive MRI contrast agents. Enhancement in these images varies based on both tissue pH and local concentration of the contrast agent. In order to generate in vivo pH maps of mice, a previously published dual-bolus technique using two contrast agents, one pH-insensitive and one pH-sensitive, was employed. Three MRI contrast agents, Gd-DTPA, Gd-DOTA-4AmP and Gd.3a, of which the latter two are pH-sensitive, were utilized. The pH-insensitive Gd-DTPA has renal pharmacokinetics comparable to the other gadolinium chelates, and thus, was utilized as a marker of the contribution of local concentration to image enhancement in the images acquired using the pH-sensitive agents. pH measurements of wild-type animals under baseline and drug-treated conditions will be presented, the applicability of a two-compartment model for describing renal physiology will be assessed, and hypotheses for relating differences in pH regulation to functional consequences of AQP1 deficiency will be discussed.

RAMOS, Tercio Carneiro. Avaliação de eficácia e segurança da acetazolamida na doença periodontal experimental. 2010. 121 f. Tese (Doutorado em Farmacologia) - Universidade Federal do Ceará. Faculdade de Medicina, Fortaleza, 2010.
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Periodontal diseases are the main cause of tooth loss in adults. The preponderant etiologic factor for the onset of periodontitis is the accumulation of dental biofilm with predominance of anaerobic gram negative bacteria. In spite of being of bacterial origin, the inflammatory response induced by factors in the host may influence the progression and clinical characteristics of periodontal disease. Alveolar bone is typical in the development of this pathology and is dependent on osteoclastic activity. In turn, one of the mechanisms of bone resorption by these cells is through the production of protons released in the resorption vacuoles by carbonic anhydrase activity. It has been reported that carbonic anhydrase inhibitors, such as acetazolamide, have a suppressive action on osteoclasts. The aim of this study was to analyze the effect of acetazolamide (ACTZ) on bone loss induced in the experimental periodontitis model in rats, as well as the dosage of seric biomarkers in the periodontal inflammatory process and histologic analysis of the periodontium. Safety analyses were also performed by means of a complete hemogram, biochemical tests for TGP, TGO, gama-GT and histopathology of organs such as: the liver, kidneys, spleen, heart and lungs. To evaluate alveolar bone loss, 50 rats received nylon ligatures in the maxillary left second molar region. The animals were divided into six groups: Vehicle Group (propylenoglycol) (n=9), Positive Control Group (alendronate 0.08 V (n=7), Negative Control Group (saline) (n=6) and Groups that received acetazolamide I.P. in the following doses: (8.3, 25 and 75 mg/Kg [Groups ACTZ8,3, ACTZ25 and ACTZ75 respectively] n= 7, 9 and 9). After 11 days, the rats were sacrificed. Alveolar bone loss was macroscopically evaluated by means of the area of root exposure. In another four groups, ACTZ25 (n=5) and vehicle (n=5), the periodontium was histologically analyzed after 11 and 17 days of ligature, by HE staining. Seric biomarkers were dosed (IL-1, IL-4, fractalkine, CINC-2, CINC-3, LIX, GM-CSF, βNGF, VEGF and CNTF) by means of the microarray technique before and after ligature in three groups; ACTZ25, Vehicle and Sham (simulated surgery). In the ACTZ25 group, safety analysis was performed (alpha=5%). Macroscopically, the alendronate Group (positive control) presented the lowest bone loss, followed by Groups ACTZ75 and ACTZ25 (p<0.05) when compared with the vehicle and saline controls, which did not differ between them (p>0.05). Histologically, the ACTZ group presented the lowest bone and cement resorption (p<0.05) after 17 days of ligature, and there was no difference between the ACTZ25 and vehicle groups with regard to inflammatory infiltrate (P>0.05). The highest concentrations of IL-4 and CNTF were observed in Group ACTZ25 (p< 0.05) when compared with the vehicle group. The safety analyses demonstrated that acetazolamide at the dose of 25 mg/Kg was well tolerated without significant alterations in the hemogram, hepatic enzymes and histopathology of the researched organ. It was concluded that acetazolamide may protect the periodontium from bone resorption induced by ligature in rats, and may be associated with mediators involved in repair, such as IL-4 and CNTF, in addition to being well tolerated.
As doenças periodontais são as principais causas de perda de dentes em adultos. O fator etiológico preponderante para desencadear a periodontite é o acumulo do biofilme dental com predominância de bactérias anaeróbicas gram negativas. Apesar de ser de origem bacteriana a resposta inflamatória induzida a partir de fatores do hospedeiro pode influenciar na progressão e nas características clínicas da doença periodontal. A perda óssea alveolar é típica na evolução desta patologia e dependente da atividade osteoclástica. Por sua vez, um dos mecanismos de reabsorção óssea por estas células é através da produção de prótons liberados nos vacúolos de reabsorção pela atividade da anidrase carbônica. Tem sido relatado que inibidores desta enzima, como acetazolamida, tem ação supressora sobre osteoclastos. O objetivo deste estudo foi analisar o efeito da acetazolamida (ACTZ) na perda óssea induzida no modelo de periodontite experimental em ratos, bem como a dosagem de biomarcadores séricos do processo inflamatório periodontal e análise histológica do periodonto. Análises de segurança também foram realizadas através de hemograma completo, dosagens de TGP, TGO, gama-GT e histopatologia de órgãos como: fígado, rins, baço, coração e pulmão. Para avaliação da perda óssea alveolar 50 ratos receberam ligaduras de nylon na região do segundo molar superior esquerdo. Os animais foram divididos em seis grupos: Grupo veículo (propilenoglicol) (n=9), grupo controle positivo (alendronato 0,08 mg/Kg) (n=7), grupo controle negativo (salina) (n=9) e grupos que receberam acetazolamida via I.P. nas doses (8,3, 25 e 75 mg/Kg [grupos ACTZ8,3, ACTZ25 e ACTZ75, respectivamente] n= 7, 9 e 9). Após 11 dias, os ratos foram mortos. A perda óssea alveolar foi avaliada macroscopicamente através da área de exposição de raiz. Em outros quatro grupos, ACTZ25 (n=5) e veículo (n=5), o periodonto foi analisado histologicamente após 11 e 17 dias de ligadura por coloração em HE. Biomarcadores séricos foram dosados (IL-1, IL-4, fractalkine, CINC-2, CINC-3, LIX, GM-CSF, βNGF, VEGF e CNTF) através da técnica de microarray antes e depois da ligadura em três grupos; ACTZ25, Veiculo e Sham (cirurgia simulada). No grupo de ACTZ25 foi realizada a análise de segurança (alpha=5%). Macroscopicamente, o grupo alendronato (controle positivo) apresentou a menor perda óssea, seguido do grupo ACTZ75 e ACTZ25 (p<0.05) quando comparados aos controles veículo e salina que não diferiram entre si (p>0.05). Histologicamente, o grupo ACTZ apresentou a menor reabsorção de osso e cemento (p<0,05) após 17 dias de ligadura, não houve diferença entre o grupo ACTZ25 e o veículo quanto ao infiltrado inflamatório (P>0,05) . As maiores concentrações de IL-4 e CNTF foram observadas no grupo ACTZ25 (p< 0,05) quando comparado ao grupo veículo. As análises de segurança demonstraram que acetazolamida na dosagem de 25 mg/Kg foi bem tolerada sem alterações significativas do hemograma, nas enzimas hepáticas e histopatologia dos órgãos pesquisados. Concluímos que a acetazolamida pode proteger o periodonto da reabsorção óssea induzida por ligadura em ratos e pode estar associada a mediadores envolvidos com o reparo, como a IL-4 e CNTF, além de ser bem tolerada.

碩士
嘉南藥理科技大學
生物科技系暨研究所
98
The purpose of this study was the preparation and investigation of drug containing polymer gel beads. Different beads were obtained by polyelectrolyte complexation of chitosan, sodium alginate and calcium chloride to form interpenetration network (IPN) primary particles. The secondary particles were accomplished on the surface chitosan of the primary particles via the ionic-crosslinking with sodium tripolyphosphate. The effect of drug entrap ratio, drug distribute density, total pore volume, morphology and physical properties on drug released behavior were investigated. The results showed that the drug can prolong about 9 hr at pH2 and its release rate was consistent with T. Higuchi model.

碩士
大仁科技大學
製藥科技研究所
96
Abstract Nowadays, health foods or scientific herb medicines which contain Lutein, rhodiola rosea and acanthopanax can be generally seen. These medicines are indicated to possess antioxidational material or activity in numerous researches. In the research of pharmacokinetics which relates with lutein and acetazolamide, it has been discovered that the maximum absorbing ratio will be increased if acetazolamide is used with lutein. Which demonstrated lutein may decrease the metabolism of acetazolamide. In this research, acetazolamide will be utilized with rhodiola rosea and acanthopanax which are the traditional medicines have antioxidational efficiency. The experiment of antioxidational analysis, such as DPPH and Xanthine Oxidase, will be performed to discuss the usage of the mixture of acetazolamide, rhodiola rosea and acanthopanax, and to clarify if the function of antioxidational efficiency could be increased. In addition, to discuss influences of this mixture to the liver and the kidney, in order to develop new clinical application of the utilizing of acetazolamide.

碩士
國立臺灣大學
化學工程學研究所
96
The purpose of this study was to apply the continuous supercritical anti-solvent (SAS) process to treat the active pharmaceutical ingredients (APIs)：sulfamerazine and acetazolamide. The aims are to increase dissolution rates of micronized APIs, and to obtain polymorphous APIs after the SAS processes. In this research, the effects of the continuous SAS process parameters were discussed. The operating parameters resulted in various particle sizes, particle size distributions, crystal habits, and polymorphs of the APIs. These parameters included solvent, pressure, temperature, concentration of solution, and solution flow rate of solution. They were systematically studied and the optimum operating conditions were reported. The smallest average particle of sulfamerazine was obtained as 0.61±0.38µm under the following operating conditions：solvent = acetone, P=140bar, T=35℃, solution concentration=30% sat., solution flow rate=0.5mL/min. The crystal habit changed from the original irregular lump to irregular flake. The dissolution rate of sulfamerazine increased by 2.9 times after the SAS process. The smallest average particle size of acetazolamide was obtained as 0.36±0.12µm under the following operating conditions：solvent = ethyl acetate, P=100bar, T=35℃, solution concentration=90% sat., solution flow rate=1mL/min. The crystal habit changed from the original irregular lump to bar shape. The dissolution rate of acetazolamide increased by 4.4 times after the SAS process. The original acetazolamide was Form II while polymorph Form I was received from the SAS process using ethanol as the solvent.

碩士
國立陽明大學
急重症醫學研究所
107
Background: The mechanisms of acetazolamide (ACZ) in prophylaxis of acute mountain sickness (AMS) remains unclear. This study evaluated the changes in physiological variables of sleep and heart rate variability (HRV) in subjects with previous history of AMS underwent prophylactic treatment of ACZ. Methods: Nonacclimatized healthy subjects were transported using a bus from 555 m to 3150 m within 3 hours. Polysomnography (PSG) was performed 3 days before ascent (T0), for two consecutive nights at 3150 m (T1 and T2), and 2 days after descent (T3). HRV was measured before sleep and after awakening from T0 to T3. AMS was diagnosed using a self-reported Lake Louise score questionnaire. Subjects confirmed to have AMS were enrolled in this study. The physiological variables and HRV were compared in AMS subjects without (control group) and with prophylactic ACZ (prophylactic ACZ group). Results: Thirteen AMS subjects were enrolled. The PSG results were analyzed in 8 and HRV were analyzed in 9 of the 13 subjects. The prophylactic use of ACZ in the subjects with a history of AMS significantly improved sleep efficiency (p = 0.012) and awakening percentages (p = 0.017) at T1, significantly higher levels of arterial oxygen saturation (SaO2) and lower values of end-tidal carbon dioxide tension (PETCO2) at four time points. Furthermore, they had a higher rapid eye movement sleep percentage (p = 0.05) at T2. Prophylactic ACZ treatment significantly increased the normalized unit of high frequency (HFnu) at T1 after awakening (p = 0.028). Conclusion: Significantly higher quality of sleep, higher SaO2 during sleep and lower PETCO2 at high altitude were found in the subjects with a history of AMS using prophylactic ACZ before rapid ascent. ACZ may accelerate the acclimatization process for rapid ascents to high altitudes by increasing parasympathetic tone based on HRV analyses.

Increase in life quality is partially linked to the use of pharmaceuticals. These are used in several areas and represent a great advancement in disease treatment and population welfare. Nevertheless, the increase in pharmaceutical usage lead to an increase in the detection of these compounds in the water courses. This happens because wastewater treatment plants are unable to fully remove these contaminants. From the most detected classes in the environment are non-steroid anti-inflammatory drugs (NSAIDs). NSAIDs are used as analgesics, anti-inflammatory and antipyretic. An example of this class is salicylic acid (SA) which similar to this class acts by inhibiting the activity of cyclooxygenase (COX). Which, in its turn is responsible for the synthesis of inflammatory mediators. From an environmental point of view, another class of interest, is that of the diuretics, particularly the subgroup of carbonic anhydrase inhibitors. Pharmacologically, this class acts by inhibiting the activity of enzyme carbonic anhydrase (CA). This enzyme is responsible for acid-base balance in organisms as well as key processes such as carbon obtention in plants, calcium mobilization in arthropods among others. The goal of this work was to evaluate the effects of environmental realistic concentrations of ACZ and SA (individually and in mixture) in non-target species (namely Lemna gibba, Mytilus spp.,and Phorcus lineatus) using biomarkers (enzymes such as CA, COX), photosynthetic pigments and biometrics. Photosynthetic pigments from L. gibba showed ACZ capacity to inhibit their synthesis. Nevertheless, in mixture with SA, these effects were reversed, showing the phyto-protection capacity of SA. In organisms from the genus Mytilus spp. exposure to ACZ display the capacity of this pharmaceutical to inhibit CA, especially in gills. When in mixture, the decrease in COX activity suggest ACZ modulates SA excretion. Finally, in organisms from P. lineatus the here tested pharmaceuticals did not caused any straightforward pattern in evaluated enzymes. In conclusion, pharmaceuticals can have different effects in different organisms. Being pharmaceutical mixture here tested capable of being beneficial to plants (by reversed photosynthetic pigment reduction) or be harmful in mussels (increased inhibition of COX activity).
O aumento da qualidade e da esperança média de vida têm sido em parte relacionados com o uso de fármacos. Estes são usados nas mais variadas áreas de atividade humana, e representam um grande avanço no tratamento de doenças e no bem-estar das populações. Contudo, este aumento do uso de fármacos tem levado a um aumento da frequência de deteção dos mesmos em ecossistemas aquáticos. Isto deve-se, entre outras razões, à incapacidade das estações de tratamento de águas residuais de removerem na totalidade estes compostos. De entre as classes mais usadas e encontradas no ambiente, podem identificar-se os anti-inflamatórios não esteroides (AINEs), que correspondem a medicamentos largamente usados a nível global como analgésicos, anti-inflamatórios e antipiréticos. Um exemplo desta classe é o ácido salicílico. À semelhança dos restantes compostos da classe onde se insere, o ácido salicílico é um inibidor da enzima cicloxigenase (COX), que é responsável pela síntese de mediadores de inflamação e pirogénicos, nomeadamente prostaglandinas e tromboxanos. Do ponto de vista ambiental, existe uma outra classe de fármacos que é igualmente importante, mas para a qual não existe um manancial de dados muito extenso. Esta classe, a dos diuréticos, caracteriza-se não pela sua abundância, mas pela sua capacidade de afetar pontos críticos em vias metabólicas de organismos não alvo, ou pela possibilidade de modelar a toxicidade de outros fármacos. De entre os diuréticos mais importantes, destacam-se particularmente os do subgrupo dos inibidores da anidrase carbónica, enzima que é responsável pelo equilibro ácido-base nos organismos bem como outros processos chave, entre eles a fixação de carbono pelas plantas, e a disponibilidade de cálcio em artrópodes. O objetivo deste trabalho foi avaliar os efeitos de concentrações ambientalmente relevantes de acetazolamida (ACZ) e do ácido salicílico (SA) (individualmente e em mistura binárias) em organismos não-alvo (nomeadamente Lemna gibba, Mytilus spp. e Phorcus lineatus) usando marcadores bioquímicos (enzimas como a CA, COX), dados biométricos e pigmentos fotossintéticos. Dados de pigmentos fotossintéticos, nomeadamente clorofilas, determinados em L. gibba mostraram a capacidade da ACZ em inibir a sua síntese. No entanto em mistura com o SA, estes efeitos foram revertidos, mostrando o potencial de fito-proteção deste fármaco. Em indivíduos do género Mytilus, exposição a ACZ demonstrou a capacidade deste fármaco em inibir a CA principalmente nas brânquias. Por outro lado, a exposição à mistura de ambos os fármacos levaram a um aumento da inibição da COX no manto, sugerindo uma modulação pela ACZ na permanência e excreção do SA em organismos deste género. Finalmente, indivíduos da espécie Phorcus lineatus não registaram alterações nos padrões de biomarcadores avaliados. Em conclusão, os fármacos (nomeadamente SA e ACZ) podem ter efeitos muito diferentes em organismos distintos, sendo que a mistura dos fármacos aqui testados parece ser benéfica na planta (invertendo a redução dos conteúdos de pigmentos fotossintéticos causados pela exposição a ACZ) e prejudicial no mexilhão (a co-exposição aos dois compostos acentuou a inibição da COX). Assim podemos concluir que os efeitos causados por fármacos podem variar de organismo para organismo bem como na extensão dos danos causados pela interação/modulação de vários contaminantes. O que pode levar a alterações nos indivíduos o que pode provocar alterações na função ecológica das espécies.
Mestrado em Eco-toxicologia e Análise de Risco

The thesis entitled “High-Resolution Charge Density Studies on Electronic Nature of Weak Interactions and Correlation of Molecular Conformation with Packing in Solid State” consist of five chapters. Chapter 1 is a brief introduction to the methodologies and techniques utilized in modelling electron densities and the topics relevant to the work. The subsequent four chapters are divided into two parts-Part A and Part B. Part A has two chapters that discusses the electronic nature of unexplored weak intermolecular interactionspnicogen bonding in nitrogen atom and hydrophobic interactions between methyl groups in molecular crystals. Part B also contains two chapters that investigates the symbiotic relation of molecular conformation and packing in solid state in two unique cases-hybridized induced polymorphism observed in sulfa drug acetazolamide and unusual asymmetry observed in overcrowded Octachloronaphthalene molecule Part A: Electronic Nature of Weak Interactions Chapter 2 discusses the electron density features of pnicogen bond between nitrogen as an electrophile and chlorine as a nucleophile from experimental and theoretical charge density analyses of 2-amino-5-nitropyridine and chloroacetic acid complex. The charge transfer nature of pnicogen bonding due to overlap between donor lone pair orbitals of Cl atom and antibonding N-C sigma star orbital has been demonstrated from gas phase NBO calculations. Presence of sigma hole on N atom is further confirmed from 3D deformation maps and electrostatic maps. Topological description from AIM analysis and energy estimation based on EML method proves this interaction to be weak in nature and comparable to the strength of carbon bonding, type II F•••F halogen bonding. Detailed Cambridge Structural Database (CSD) analysis reveals that planar N atoms have the maximum propensity to participate as electrophile in pnicogen bonding. Chapter 3 reports the frequent occurrence of methyl•••methyl hydrophobic interactions in the solid-state from CSD study with a detailed analysis of these interactions in a series of cocrystals of biologically active molecules such as caffeine, theophylline and tetramethylpyrazine using experimental X-ray charge density analysis, variable-temperature crystallography and solidstate NMR. The visualization of accurate electron density distribution in the interaction region reveals that they are stabilized by the minimized electrostatic repulsion and maximized dispersion forces. This chapter further proves methyl•••methyl HI as a group•••group interaction with a pronounced torsional vibration for the hydrophobic methyl groups which leads to a significant entropic contribution towards its stability. A characteristic C-13 ssNMR up-field chemical shift was found to be associated with these methyl•••methyl interactions in the crystal state. Part B: Correlation of Molecular Conformation with Packing Chapter 4 discusses a new type of polymorphism called hybridized induced polymorphism in connection with the unusual phenomenon of the formation of kinetic form as against the thermodynamic form on slow cooling of boiling aqueous solution of diuretic drug Acetazolamide. Experimental charge density analysis aided with ab initio calculations have investigated the local electron density at the amino region of both polymorphs. A series of crystallization experiments of AZM in aqueous medium were conducted. The boiling solution was ramped down at different rates of cooling; rapid cooling in liquid nitrogen, ambient cooling to room temperature, controlled cooling at (10°C/hr, 7°C/hr and 5°C/hr) to room temperature. PXRD analysis reveals the kinetic form occurs only when the cooling rate is quite slow (7°C/hr and 5°C/hr). The occurrence of both polymorphs from aqueous solution of AZM under different crystallization conditions is rationalized in terms of hybridization induced polymorphism. Chapter 5 investigates electron density distribution in an overcrowded aromatic molecule, Octachloronaphthalene (OCN) by charge density analysis to unravel several unexplored factors responsible for steric hindrance. The topological features of the enigmatic peri interactions contributing to steric overcrowding are qualified and quantified from experimental and theoretical charge density studies. A new facet in the fundamental understanding of peri interactions is revealed by NCI (Non-Covalent Interaction) analysis. The potential role of these interactions in deforming the molecular geometry and subsequent effect on aromaticity are substantiated from NICS (Nuclear Independent Chemical Shift) and QTAIM (Quantum Theory of Atoms in Molecules) calculations. The eye-catching dissimilarity in the out-of-plane twisting of OCN renders the molecule in an asymmetric geometry in the crystalline phase as compared to symmetric geometry in the optimized solvated phase. This is uniquely characterised by their molecular electrostatic potential (MESP) respectively and is explained in terms of conflict between two opposing forces- peri interactions and symbiotic intermolecular Cl•••Cl and Cl••• contacts.

The aim of this thesis was to elucidate the role of carbonic anhydrase (CA) in the modulation of central pH/CO2-sensitive fictive breathing (measured using in vitro brainstem-spinal cord preparations) in leopard frogs (Rana pipiens) following exposure to chronic hypercapnia (CHC) and chronic hypoxia (CH). CHC caused an augmentation in fictive breathing compared to the controls (normoxic normocapnic). Addition of acetazolamide (ACTZ), a cell-permeant CA inhibitor, to the superfusate reduced fictive breathing in the controls and abolished the CHC-induced augmentation of fictive breathing. ACTZ had no effect on preparations taken from frogs exposed to CH. Addition of bovine CA to the superfusate did not alter fictive breathing in any group, suggesting that the effects of ACTZ were due to inhibition of intracellular CA. Taken together, these results indicate that CA is involved in central pH/CO2 chemoreception and the CHC-induced increase in fictive breathing in the leopard frog.

Dissertação de mestrado em Tecnologias do Medicamento, apresentada à Faculdade de Farmácia da Universidade de Coimbra
A deficiência visual que muitas vezes leva à cegueira está entre as doenças que causam maior taxa de morbidade. Os erros refrativos, o glaucoma, as cataratas e as infeções são alguns exemplos. O objetivo deste trabalho é criar um implante biodegradável obtido a partir de fibras coaxiais contendo o fármaco disperso – acetazolamida. O implante será inserido no segmento posterior do globo ocular do doente em contexto de cirurgia de ambulatório e o fármaco será libertado gradualmente, não necessitando de injeções periódicas, ao contrário das injeções intravítreas de esteroides. Durante este trabalho desenvolveram-se, em primeiro lugar, implantes sem revestimento, compostos por membranas fibrosas produzidas por electrospinning e enroladas na forma de pequenos cilindros. O fármaco foi imobilizado no interior das fibras durante a sua preparação, sendo estas fibras compostas por PCL (policaprolactona) e lutrol F127 (poli (oxietileno-b-oxipropileno-b-oxietileno)). Produziram-se fibras com uma estrutura uniaxial (compostas pela mistura de PCL, lutrol F127 e acetazolamida) e com uma estrutura coaxial (com um núcleo de lutrol F127 com acetazolamida e um revestimento de PCL). Os implantes preparados apresentaram uma velocidade de libertação de fármaco muito rápida, o que levou à preparação de novos implantes, incorporando as mesmas formulações que os primeiros, mas com a adição de um filme de revestimento, preparado por solvent casting e composto por PCL e lutrol F127 ou por PCL e luwax EVA 3 (poli (etileno-co-acetato de vinilo)). Os implantes e os seus constituintes (membranas fibrosas e filmes de revestimento) foram caracterizados por várias técnicas, tais como: SEM, FTIR-ATR, análise térmica simultânea e determinação de ângulos de contacto. Adicionalmente, procedeu-se à obtenção dos perfis de libertação in vitro da acetazolamida, em PBS e a 37ºC, utilizando um método espetrofotométrico para quantificar o fármaco libertado ao longo do tempo. Os resultados mostraram que o perfil de libertação da acetazolamida a partir dos implantes com revestimento é consideravelmente mais lenta e gradual, dado que estes sistemas apresentam uma barreira adicional à difusão do fármaco. No caso do revestimento contendo lutrol F127, polímero hidrofílico, ocorreu intumescimento do revestimento diminuindo a taxa de libertação do fármaco. No que se refere ao revestimento contendo luwax EVA 3, polímero hidrofóbico, a sua degradação por erosão permitiu diminuir a taxa de libertação do fármaco.
The visual impairment that often leads to blindness is among the diseases that cause higher morbidity rate. Refractive errors, glaucoma, cataracts and infections are some examples. The objective of this work is to create a biodegradable implant obtained from coaxial fibers containing the dispersed drug - acetazolamide. The implant is inserted into the posterior segment of the eye globe in the patient in ambulatory surgery context and the drug is released gradually, not requiring periodic injections, unlike intravitreal steroids injections. During this work uncoated implants were developed at first, they were composed of fibrous membranes produced by electrospinning and rolled in the shape of small cylinders. The drug was immobilized inside the fibers during their preparation and these fibers were composed of PCL (polycaprolactone) and lutrol F127 (poly (oxyethylene-b-oxypropylene-boxyethylene)). Fibers were produced with a uniaxial structure (consisting of the mixture of PCL, lutrol F127 and acetazolamide) and with a coaxial structure (lutrol F127 and acetazolamide as core and PCL as shell). The prepared implants exhibited a fast rate of drug release, which led to the preparation of new implants incorporating the same formulation as the first but with an additional coating film prepared by solvent casting and comprising PCL and lutrol F127 or PCL and luwax EVA 3 (poly (ethylene-co-vinyl acetate)). Implants and their constituents (fibrous membranes and coating films) were characterized by various techniques, such as: SEM, FTIR-ATR, simultaneous thermal analysis and determination of contact angles. In addition, in vitro release profiles of acetazolamide were obtained in PBS at 37° C using a spectrophotometric method for quantifying the drug released over time. The results have showed that the release profile of the acetazolamide of coated implant is considerably slower and more gradual, since the drug has to penetrate an additional barrier. In case of coating containing lutrol F127, an hydrophilic polymer, the swelling of the coating led to a slow rate of drug release. In what concerns the coating containing luwax EVA 3, an hydrophobic polymer, its degradation by erosion allowed a slow rate of drug release.