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Previous issue date: 2016-02-15 / Nuclear Medicine enables, among other things, the visualization, characterization and quantification of biochemical processes, metabolism, biomarkers and receptors in vivo by molecular imaging. One of the methods of functional imaging available in this area is Positron Emission Tomography (PET), which depends on the administration of a radiopharmaceutical in the studied organism. One disease that can be studied by PET scan is multiple sclerosis (MS) through the access to the TSPO (Translocator protein) receptors density. The radiopharmaceutical [11C]-(R) -PK11195 has high affinity and selectivity for these receptors, which are increased in activated microglia cells in the case of MS. Therefore, the objective of this study was to develop and to validate the production process (synthesis, purification and quality control) of the radiotracer [11C]-(R)-PK11195 in the Radiopharmaceutical Production Center of the Brain Institute of Rio Grande do Sul (BraIns), allowing performing PET scans for the study of MS. The radioisotope was produced in the 16 MeV PETrace cyclotron (GE Healthcare) as [11C]CO2 via the nuclear reaction 14N (p, ?)11C after the bombardment of a gaseous target containing N2 and 0.5% of O2 with protons. The production was performed in the equipment TRACERlab FX-C Pro (GE Healthcare) to meet the requirements of Good Manufacturing Practices (GMP). The radiopharmaceutical quality control was carried out at the BraIns Quality Control Laboratory and the final product was tested for Identity and Radionuclidic Purity, Radiochemical Purity, Specific Activity, Residual Solvents, pH, Sterility, the Filtration Membrane Integrity and Bacterial Endotoxins. The synthesis was performed through the methylation of the precursor (R)-[N-desmethyl]PK11195 using [11C]CH3I (methyl iodide labeled with carbon-11) under the defined optimal conditions: 400 uL of dimethyl sulfoxide (DMSO), approximately 1 mg of precursor, approximately 10 mg of potassium hydroxide (KOH) at 40 ?C for 1 minute. The reaction mixture was then neutralized with 1 mol.L-1 hydrochloric acid and diluted with mobile phase (76% acetonitrile and 24% Milli-Q water). The diluted mixture was injected into a semi-preparative HPLC system for purification. Solid phase extraction (through Sep-Pak C18 cartridges - Waters) was used to remove the solvent. Product elution and formulation was conducted with 0.7 mL of ethanol and 6.3 ml of 0.9% saline (sterile and pyrogen free). Sterilization was performed by filtration on a 0.22 ?m pore membrane and, as well as the filling process, was carried out under laminar flow. The process steps could be optimized and fitted to the equipments and laboratories structure. Radiochemical yield of the process in relation to the [11C ]CH3I start activity was 15.5 ? 2.4 % ( 7471.5 ? 1283.8 MBq ; n = 39 ) . All batches were approved according to the specifications defined and tested in the quality control, reaching Radiochemical Purity levels greater than 99% and a Specific Activity of 39.7 ? 11.1 GBq / micromol , 10 minutes before the injection. All analytical methods were validated and met the requirements established by current regulations. Three consecutive batches were produced and approved by the quality control, so the process could be considered validated. The developed process was suitable for production of adequate doses of the radiopharmaceutical [11C]-(R)-PK11195 for clinical use at BraIns. The production of the radiopharmaceutical labeled with carbon-11 at the institute provides an additional tool for improving health services in the country and to improve the life quality of the patients. / A medicina nuclear possibilita, dentre outras coisas, a visualiza??o, caracteriza??o e quantifica??o de processos bioqu?micos, metab?licos, biomarcadores e receptores, in vivo, atrav?s da imagem molecular. Dentre as modalidades de imagem funcional dispon?veis nesta ?rea est? a Tomografia por Emiss?o de P?sitrons (PET), que depende da administra??o de um radiof?rmaco no organismo estudado. Uma das doen?as pass?veis de estudo atrav?s do exame PET ? a esclerose m?ltipla (EM), atrav?s do acesso ? densidade de receptores TSPO (Prote?na Translocadora). O radiof?rmaco [11C]-(R)-PK11195 possui alta afinidade e seletividade por estes receptores, aumentados nas c?lulas ativadas da micr?glia nos casos de EM. Portanto, o objetivo deste trabalho foi desenvolver e validar o processo produtivo (s?ntese, purifica??o e controle de qualidade) do radiof?rmaco [11C]-(R)-PK11195 no Centro de Produ??o de Radiof?rmacos do Instituto do C?rebro do RS (InsCer), possibilitando a realiza??o de exames PET para o estudo da EM. O radiois?topo foi produzido no C?clotron PETrace 16 MeV (GE Healthcare) na forma de [11C]CO2 atrav?s da rea??o nuclear 14N(p,?)11C, por meio do bombardeamento com pr?tons do alvo gasoso contento N2 e 0,5 % de O2. A produ??o foi realizada no equipamento TRACERlab FX-C pro (GE Healthcare) atendendo ?s exig?ncias de Boas Pr?ticas de Fabrica??o (BPF). O controle de qualidade do radiof?rmaco foi realizado no Laborat?rio de Controle de Qualidade do InsCer e o produto acabado foi submetido ?s an?lises de Identidade e Pureza Radionucl?dica, Pureza Radioqu?mica, Atividade Espec?fica, Solventes Residuais, pH, Esterilidade, Integridade da Membrana Filtrante e Teor de Endotoxinas. A s?ntese foi realizada atrav?s da metila??o do precursor (R)-[N-desmetil]PK11195 utilizando o [11C]CH3I (iodeto de metila marcado com carbono-11) sob as condi??es ?timas definidas: 400 ?L de dimetilsulf?xido (DMSO), aproximadamente 1 mg de precursor, aproximadamente 10 mg de hidr?xido de pot?ssio (KOH), a 40 ?C durante 1 minuto. A mistura de rea??o foi posteriormente neutralizada com ?cido clor?drico 1 mol.L-1 e dilu?da com fase m?vel (acetonitrila 76 % e ?gua Milli-Q 24 %). A mistura dilu?da foi injetada em sistema CLAE semi-preparativo para purifica??o. Extra??o de fase s?lida (atrav?s de cartuchos SepPak C18 ? Waters) foi utilizada para remo??o do solvente. A elui??o e formula??o do produto foi realizada com 0,7 mL de etanol e 6,3 mL de solu??o salina 0,9 % (est?ril e livre de pirog?nios). A esteriliza??o foi realizada atrav?s de filtra??o em membrana de poro de 0,22 ?m e, assim como o envase, ocorreu em fluxo laminar. As etapas do processo foram otimizadas e adequadas aos equipamentos e estrutura dos laborat?rios. O rendimento radioqu?mico do processo em rela??o ? atividade de [11C]CH3I de partida foi de 15,5?2,4 % (7471,5?1283,8 MBq; n=39). Todos os lotes produzidos foram aprovados quanto ?s especifica??es definidas e testadas no controle de qualidade, atingindo Pureza Radioqu?mica maior que 99 % e uma Atividade Espec?fica de 39,7?11,1 GBq/?mol, 10 minutos antes da inje??o. Todas as metodologias anal?ticas foram validadas e atenderam os crit?rios estabelecidos pela norma vigente. Tr?s lotes consecutivos foram produzidos e aprovados no controle de qualidade e o processo p?de ser considerado validado. O processo desenvolvido se mostrou adequado para produ??o do radiof?rmaco [11C]-(R)-PK11195 em doses suficientes para realiza??o de exames em pacientes no InsCer. A produ??o do radiof?rmaco marcado com carbono-11 no instituto disponibiliza mais uma ferramenta para melhoria dos servi?os de sa?de no pa?s e para melhoria da qualidade de vida dos pacientes.
| Identifer | oai:union.ndltd.org:IBICT/oai:tede2.pucrs.br:tede/6855 |
| Date | 15 February 2016 |
| Creators | Alba, Marcos Vin?cius Fortes |
| Contributors | Jeckel, Cristina Maria Moriguchi |
| Publisher | Pontif?cia Universidade Cat?lica do Rio Grande do Sul, Programa de P?s-Gradua??o em Biotecnologia Farmac?utica, PUCRS, Brasil, Faculdade de Farm?cia |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da PUC_RS, instname:Pontifícia Universidade Católica do Rio Grande do Sul, instacron:PUC_RS |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | 2304961219518893267, 600, 600, 600, -8380654636843378116, 6997636413449754996 |
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