<p> Nuclear medicine requires the use of radio labelled pharmaceuticals in order to carry out imaging and therapeutic protocols.1 Unfortunately, traditional radiolabelling approaches used in radiopharmaceutical synthesis often generate multiple products, which require exhaustive HPLC purification prior to use.2 Chromatographic purification reduces radiochemical yields, increases exposure, and can, in certain cases, preclude the use of shorter-lived isotopes.</p> <p> In light of the limitations of current radio labelling methods, we endeavored to develop a versatile and efficient radiolabelling strategy that would avoid the need for HPLC purification. To this end, the compounds to be labelled were first bound to a highly fluorinated stannylated precursor, which, when reacted with a radiohalogen, generated the corresponding radiolabelled compound. Unlike other halodestannylation reactions, however, the radiolabelled compound could be isolated from the stannylated precursor by elution through a fluorous Sep-Pak.</p> <p> As a model system, tris(perfluorohexylethyl)tin-3-benzoic acid (1), was synthesised and labelled. Compound 1 was prepared through a novel reaction which involved treatment of bromotris[(2-perfluorohexyl)ethyl]tin with the organozinc reagent 3-(ethoxycarbonyl)phenylzinc. Reaction of compound 1 with [18F]F2, followed by fluorous Sep-Pak purification, generated the corresponding labelled 3-[18F]fluorobenzoic acid in 27 min, in 30% radiochemical yield, and having a specific activity of 1966 mCi/mol. Alternatively, reaction of compound 1 with Na125I provided the corresponding product, 3-[125I[iodobenzoic acid, in <1 hr, in 75% radiochemical yield, and greater than 99% radiochemical purity. Prior tests clearly showed that any excess or unreacted substrate was fully removed from the product using a fluorous Sep-Pak.</p> <p> In addition to the initial validation studies, new synthetic methods were developed as a means of preparing more complex "fluorous" substrates. A coupling methodology was developed which permitted synthesis of a fluorous "tagged" benzamide, through reaction of 1 in the presence of HBTU with N,N-dimethylethylenendiamine. Subsequent labelling using F2 and I2 has been shown to generate the corresponding labelled benzamides, which are important agents for imaging melanoma and dopamine receptors.3 Another relevant radiopharmaceutical precursor, tris[(2-perfluorohexyl)ethyl]tin-3-benzylamine (2), was synthesized though reaction of bromotris[(2-perfluorohexyl)ethyl]tin with 1-(3-bromobenzyl)-2,2,5,5-tetramethyl-1,2,5-azadisilolidine. Compound 2 was successfully coupled to the chemotactic peptide, GFLM(f), and the product subsequently labelled with iodine. Compound 2 was also used to prepare the corresponding benzylguanidine (3), an important precursor to m-iodobenzylguanidine, which is used for imaging and therapy of neural crest tumors.4 Initial labelling results show that reaction of 3 with NaI and an oxidant generates the corresponding labelled m-iodobenzylguanidine.</p><p> Results suggest that the fluorous synthesis method will offer several advantages over traditional radiolabelling strategies. The radiolabelled products are generated in high yield, through rapid and facile reactions that avoid the need for HPLC purification.</p> / Thesis / Master of Science (MSc)

Magister Pharmaceuticae - MPharm / The purpose of this research thesis, is to evaluate the appropriateness of the current regulatory guidelines on the manufacture of medicinal products within the radiopharmaceutical industry. The manufacture of radiopharmaceuticals is governed by two main regulatory bodies. The regulatory authority responsible for the governance of the pharmaceutical product has the primary objective of ensuring the safety, efficacy and purity of the products manufactured, while the nuclear regulator has the responsibility of ensuring that the products used and manufactured are done so in a safe and responsible manner. The main problem faced by this industry is one where the requirements between the two regulators are, in some instances, in direct conflict of each other. The methodology employed in this study included the review of current regulatory guidelines applicable to the manufacture of orthodox pharmaceuticals, compared to its applicability to the radiopharmaceutical industry. This review showed that the regulatory guidelines, when adopted without industry specific concessions, resulted in an industry which is not as appropriately regulated as orthodox medicines are. It is therefore concluded that, in order for the radiopharmaceutical manufacturing industry to be as appropriately regulated as the orthodox pharmaceutical industry, industry-specific guidelines are required to be developed by the regulatory authorities and adopted by the manufacturing industry.

Thesis (MScMedSc)--Stellenbosch University, 2010. / ENGLISH ABSTRACT: Radiolabelled compounds have been widely used as investigative tools for psychiatric disorders using positron emission tomography (PET) or single photon emission tomography (SPECT) of the brain. In particular 123I-5-IR91150, a serotonin (5-HT) 2a antagonist, has been used for imaging the serotonergic system. The current study developed optimal radiolabelling and purification methods in our laboratory with the objective that it can provide 123I- 5-I-R91150 in sufficient quantity and of acceptable pharmaceutical quality for human use. Unlabelled R91150 was obtained from Janssen Pharmaceutica (Beerse, Belgium). Carrier free [123I]Iodine was produced by iThemba LABS, South Africa, via the 127I(p,5n)123Xe-123I reaction, providing Na[123I] in 0.05 N sodium hydroxide with a specific activity of 4000-6000 MBq/ml. A direct electrophilic radioiodination method of labelling was used in this study for labelling 123-I-5-IR91150 in glacial acetic acid. After radiolabelling, the product was purified using two different methods, namely a high performance liquid chromatography (HPLC) purification method and a solid phase extraction (SPE) method. The analyses of the purified product for both methods were done using HPLC. Methods were tested to reduce the volume of the purified product using C8 or C18 solid phase extraction cartridges. The average labelling efficiencies for SPE and HPLC purification methods were 76% ± 13.6% and 52% ± 11.2% respectively. The yields of 123I-5-I-R91150 were about 80%. Sep-Pak C8 and C18 were both unable to concentrate the HPLC purified product. Products from both purification methods were sterile and pyrogen free. Both SPE and HPLC purification methods have been shown to provide products meeting most criteria set for this study. However, both methods have advantages and disadvantages. The SPE purification method provided higher labelling efficiency and a much lower product volume. The stability of this product is however of concern as some free iodide was detected. If this purification method is used, the product should therefore be administered as soon as possible after completion of analysis. After HPLC purification, the undiluted product remained stable up to 4.15 hours after production but the product volume was relatively high, and purification time-consuming. In order to obtain a useful patient dose, labelling would have to start with at least 740 MBq 123I and the labelled product should be collected in fractions of 5 ml or less in order to obtain a fraction of sufficiently high specific activity. It was concluded that radiolabeling R91150 is possible at our institution, but that an improved HPLC system would be of value for routine production of a pure and safe product. / AFRIKAANSE OPSOMMING: Radioaktief gemerkte verbindings word baie gebruik as ondersoekmiddel vir psigiatriese afwykings met behulp van positron emissive tomografie (PET) of enkelfoton emissie tomografie (SPECT) van die brein. Die verbinding 123I-5-IR91150, ‘n serotonien (5-HT) 2a antagonis, is beskryf vir beelding van die serotonerge sisteem. Die huidige studie het ondersoek ingestel na optimale metodes vir radioaktiewe merking en suiwering vir ons laboratorium met die doel om 123I-5-I-R91150 in genoegsame hoeveelhed en van aanvaarbare farmaseutiese gehalte geskik vir menslike gebruik te verskaf. R91150 is van Janssen Pharmaceutica (Beerse, België) verkry. Draervry [123I]jodium is deur iThemba LABS, Suid-Afrika, via die 127I(p,5n)123Xe-123I reaksie geproduseer, om Na[123I] in 0.05 N natriumhidroksied met spesifieke aktiwiteit van 4000-6000 MBq/ml te lewer. ‘n Direkte elektrofiliese radioiodineringsmetode is in hierdie studie gebruik om 123-I-5-I-R91150 in ysasynsuur te merk. Na radioaktiewe merking is die radioaktiewe produk deur twee verskillende metodes gesuiwer, naamlik ‘n HPLC metode en ‘n soliede fase ekstraksie (SPE) metode. Vir beide metodes is die produk deur middel van HPLC analiseer. Metodes is getoets om die volume van die gemerkte produk met C8 of C18 SPE kolommetjies te verminder. Die gemiddelde bindingsdoeltreffendheid vir die SPE en HPLC suiweringsmetodes was 76% ± 13.6% en 52% ± 11.2% onderskeidelik. Die opbrengs van 123I-5-I-R91150 was ongeveer 80%. Sep-Pak C8 en C18 kon beide nie gebruik word om die HPLC gesuiwerde produk te konsentreer nie. Produkte van beide suiweringsmetodes was steriel en pirogeenvry. Daar is getoon dat beide suiweringsmetodes produkte lewer wat aan die meeste kriteria wat in hierdie studie gestel is, voldoen. Beide metodes het egter voor- en nadele. Die SPE suiweringsmetdode het tot hoër bindingsdoeltreffendheid gelei, asook ‘n baie laer produkvolume. Daar is egter ‘n mate van kommer oor die stabiliteit van die produk aangesien vry radiojodied waargeneem is. Indien hierdie suiweringsmetode gebruik word, moet die produk dus so gou as moontlik na voltooiing van analise toegedien word. Na HPLC suiwering was die onverdunde produk tot 4.15 uur na produksie stabiel maar die produkvolume was relatief hoog en suiwering tydrowend. Om ‘n bruikbare pasiëntdosis te verkry moet merking met ten minste 740 MBq 123I begin en die gemerkte produk moet na suiwering in fraksies van 5 ml of minder versamel word om ‘n fraksie met geskikte spesifieke aktiwiteit te verkry. Die gevolgtrekking is gemaak dat radioaktiewe merking van R91150 by ons instelling moontlik is, maar dat ‘n verbeterde HPLC sisteem vir roetineproduksie van ‘n suiwer en veilige produk van waarde sou wees.

Antes de se iniciar os estudos clínicos de uma nova droga, é necessário realizar uma bateria de testes de segurança, para avaliar o risco humano. Os radiofármacos como qualquer outra nova droga, devem ser testados levando em conta sua especificidade, duração de tratamento e principalmente a toxicidade de ambas as partes, a molécula não marcada e a sua radioatividade em si, além das impurezas provindas da radiólise. Órgãos regulatórios como o Food and Drug Administration-EUA (FDA) e a Agência de Medicina Européia (EMEA), estabelecem guias para a regulamentação de produção e pesquisas de radiofármacos, No Brasil a produção de radiofármacos não era regulamentada até o final de 2009, quando foram estabelecidas pela Agência Nacional de Vigilância Sanitária (ANVISA) as resoluções nº 63, que visa as Boas Práticas de Fabricação de Radiofármacos e a nº 64 que visa o registro do radiofámaco. Para a obtenção do registro de radiofármacos são necessárias a comprovação da qualidade, segurança, eficácia e especificidade do medicamento. Para a segurança dos radiofármacos devem ser apresentados estudos de toxicidade aguda, subaguda e crônica como também a toxicidade reprodutiva, mutagênica e carcinogênica. Hoje o IPEN-CNEN/SP produz um dos radiofámacos mais importantes da medicina nuclear, o 18F-FDG, que é utilizado em muitas aplicações clínicas, em particular no diagnóstico e estadiamento de tumores. O objetivo deste trabalho foi avaliar a toxicidade sistêmica (aguda/subaguda) do radiofármaco 18F- FDG em um sistema teste in vivo, conforme preconiza a RDC nº 64, que servirá de modelo para os protocolos de toxicidade dos radiofármacos produzidos no IPEN. Os ensaios realizados foram: os testes de toxicidade aguda e de toxicidade subaguda, estudos de biodistribuição do 18F-FDG, ensaio cometa e toxicidade reprodutiva. Na toxicidade aguda, ratos sadios foram injetados com 18F- FDG e observados durante 14 dias enquanto na toxicidade subaguda os animais foram observados durante 28 dias. Os resultados não mostraram nenhuma evidência de toxicidade na exposição ao 18F-FDG na toxicidade aguda e na subaguda. A biodistribuição demonstrou resultados semelhantes aos da literatura, onde a bexiga é o órgão que mais recebe radiação. O ensaio cometa mostrou que a radiação do radiofármaco não foi significativa para gerar danos no DNA. Na toxicidade reprodutiva, casais de ratos expostos ao 18F-FDG geraram filhotes completamente normais e saudáveis. Por fim, o 18F-FDG não evidenciou nenhuma toxicidade. / Before starting clinical trials of a new drug, it is necessary to perform a battery of safety tests for assessing human risk. Radiopharmaceuticals like any new drug must be tested taking into account its specificity, duration of treatment and especially the toxicity of both parties, the unlabeled molecule and its radionuclide, apart from impurities emanating from radiolysis. Regulatory agencies like the Food and Drug Administration - USA (FDA) and the European Medicine Agency (EMEA), establish guidelines for the regulation of production and research of radiopharmaceuticals. In Brazil the production of radiopharmaceuticals was not regulated until the end of 2009, when were established by the National Agency for Sanitary Surveillance (ANVISA) resolutions No. 63, which refers to the Good Manufacturing Practices of Radiopharmaceuticals and No. 64 which seeks the registration of record radiopharmaceuticals. To obtain registration of radiopharmaceuticals are necessary to prove the quality, safety, efficacy and specificity of the drug . For the safety of radiopharmaceuticals must be presented studies of acute toxicity, subacute and chronic toxicity as well as reproductive, mutagenic and carcinogenic. Nowadays IPEN-CNEN/SP produces one of the most important radiopharmaceutical of nuclear medicine, the 18F-FDG, which is used in many clinical applications, particularly in the diagnosis and staging of tumors. The objective of this study was to evaluate the systemic toxicity (acute/ subacute) radiopharmaceutical 18F-FDG in an in vivo test system, as recommended by the RDC No. 64, which will serve as a model for protocols toxicity of radiopharmaceuticals produced at IPEN . The following tests were performed: tests of acute and subacute toxicity, biodistribution studies of 18F-FDG, comet assay and reproductive toxicity. In acute toxicity, healthy rats were injected with 18F-FDG and observed for 14 days while in subacute toxicity animals were observed for 28 days. The results showed no evidence of toxicity at exposure 18F-FDG in acute and subacute toxicity. The biodistribution showed similar results to the literature, where the bladder is the organ that receives the most radiation. The comet assay showed that the radiation from the radiopharmaceutical was not significant to generate DNA damage. In reproductive toxicity in coupled rats exposed to 18F-FDG generated completely normal and healthy puppies. Finally, the 18F-FDG did not show any toxicity.

Os radiofármacos são fármacos que possuem em sua composição um radionuclídeo e são utilizados para diagnóstico e tratamento de diversas doenças na medicina nuclear. O principal objetivo desse trabalho foi avaliar o efeito da temperatura e umidade na estabilidade de alguns reagentes liofilizados ALBUMINA-TEC, CIS-TEC, DEX-500-TEC, DEX-70-TEC, DISI-TEC, DMSA-TEC, DTPA-TEC, FITA-TEC, PUL-TEC, MIBI-TEC, MDP-TEC, PIRO-TEC, e TIN-TEC. Foi utilizada uma câmara climática para expor amostras destes produtos a uma temperatura de 40 °C e 75% de umidade relativa por 48 horas, para simular uma condição de transporte. Foram avaliadas duas situações: após um determinado tempo de armazenamento em temperatura de 2 - 8 °C, amostras de RL foram expostas por 48 horas em 40 °C e 75% UR e em seguida foram analisadas enquanto outras amostras foram colocadas por 48 horas em 40 °C e 75% UR, mantidas em temperatura de 2 - 8 °C até a validade do radiofármaco e analisadas quanto à pureza radioquímica e biodistribuição. Os radiofármacos ALBUMINA-TEC, DEX-70-TEC, DEX-500-TEC, DISI-TEC, DMSA-TEC, DTPA-TEC, PIRO-TEC, MDP-TEC, MIBI-TEC e MDP-TEC apresentaram resultados de % PRq e biodistribuição que atenderam às especificações, enquanto que CIS-TEC, PUL-TEC e TIN-TEC apresentaram alguns valores de % PRq menor que o especificado. TIN-TEC apresentou % PRq de 84% em um de três lotes, enquanto PUL-TEC apresentou % PRq de 88% em um de quatro lotes analisados; CIS-TEC é um RL que ainda precisa melhor analisado pois os resultados fora do especificado no controle radioquímico e biológico foram obtidos com um produto fora da validade especificada. Estes resultados indicaram que a pureza radioquímica pode ser afetada pela condição de estudo, o que não foi confirmado pelo controle biológico. / Radiopharmaceuticals are pharmaceuticals that have in their composition a radionuclide and are used for diagnosis and treatment of various diseases in nuclear medicine. The main objective of this study was to evaluate the effect of temperature and humidity on the stability of some lyophilized reagents: ALBUMINA-TEC, CIS-TEC, DEX-500-TEC, DEX-70-TEC, DISI-TEC, DMSA-TEC, DTPA-TEC, FITA-TEC, PUL-TEC, MIBI-TEC, MDP-TEC, PIRO-TEC and TIN-TEC. A climatic chamber was used for exposing some samples of these products to a temperature of 40 °C and 75% RH for 48 hours to simulate a transport condition. Two situations were evaluated: after a certain storage time at 2 - 8 °C, LR samples were exposed for 48 hours at 40 °C and 75% RH and analyzed while other samples were placed for 48 hours at 40 °C and 75% RH, kept at 2 - 8 °C until the expiration time of each radiopharmaceutical and analyzed for radiochemical purity and biodistribution. ALBUMINA-TEC, DEX-70-TEC, DEX-500-TEC, DISI-TEC, DMSA-TEC, DTPA-TEC, PIRO-TEC, MDP-TEC, MIBI-TEC and MDP-TEC presented results of % RCP and biodistribution that met the specifications while CIS-TEC PUL-TEC and TIN-TEC presented some % RCP lower than specified. TIN-TEC presented % RCP of 84% in one of three lots while PUL-TEC presented % RCP of 88% in a one of four batches analyzed; CIS-TEC is a LR that further need to be analyzed because probable the out of specification results of radiochemical and biological controls were obtained with a product outside of the validity date. These results indicated that radiochemical purity, can be affected by study condition, which has not been confirmed by biological control.

Este trabalho tem como objetivo investigar a concentração de impurezas radioativas gama emissoras presentes nas soluções dos radiofármacos produzidos no Instituto de Pesquisas Energéticas e Nucleares - IPEN em São Paulo. Para que este radiofármaco possa ser utilizado adequadamente, sua qualidade deve ser avaliada de acordo com os procedimentos estabelecidos de acordo com os \"Requisitos Gerais para a Competência de Laboratórios de Teste e Calibração\", ISO / IEC 17025: 2005 e pelas \"Boas Práticas de Fabricação\" (BPF), controladas pela ANVISA (Agência Nacional de Vigilância Sanitária), no Brasil. Para determinar a atividade, dos radiofármacos das impurezas gama emissoras, foi utilizado um espectrômetro gama de alta resolução em duas distâncias fonte-detector; uma de 18 cm e outra de 1,7 cm. Para a distância de 18 cm, o espectrômetro HPGe foi calibrado com energias gama entre 81 kev e 1408 kev, medindo ampolas seladas de 60Co, 133Ba, 137Cs e 152Eu, padronizadas no Laboratório de Metrologia Nuclear (LMN) do IPEN. Para impurezas com baixas atividades, utilizou a distância fontedetector de 1,7 cm. A esta distância, o efeito soma em cascata é muito elevado, tornando difícil a medição das ampolas de calibração padrão, com isso, a curva de eficiência do espectrômetro foi obtida por um código de simulação de Monte Carlo, desenvolvido no IPEN. Neste código, todos os detalhes do sistema de detecção são modelados e as curvas de resposta para raios X e raios gama são calculadas pelo código de transporte de radiação MCNPX. Os espectros gama foram analisados pelo programa Alpino, que aplica o método de integração numérica da área sob os fotopicos de absorção total. Para as impurezas gama emissoras não detectadas visualmente, os limites de detecção foram calculados a partir da taxa de contagem de fundo, sob a área do pico de interesse. As soluções radioativas analisadas foram 67Ga,99Mo, 99mTc, 111In, 131I, 153Sm, 177Lu e 201Tl. Os resultados da relação entre a atividade do radionuclídeo em análise e as impurezas identificadas apresentaram acordo com os certificados de análise dos fabricantes, assim como, com as especificações da ANVISA. / This work aims to investigate the concentration of radioactive impurities gamma emitters in the radiopharmaceutical solutions produced at Nuclear and Energy Research Institute -IPEN in São Paulo, So that this radiopharmaceutical may be used properly, its quality should be evaluated in accordance with the procedures established by quality control agencies, such as \"General Requirements for the Competence of Testing and Calibration Laboratories\", ISO/IEC 17025:2005 and the \"Good Laboratory Practice\" (GLP), controlled by ANVISA (National Agency Health Surveillance), in Brazil, requiring a confirmation of the values of impurities related at the certificates supplied by the manufacturers. To determine the activity, a high resolution gamma spectrometer were used in two source-detector distances. One was 18 cm and the other 1.7 cm. For the 18 cm distance, the high pure germanium spectrometer was calibrated in the energy range between 81 keV and 1408 keV by measuring sealed ampoules of 60Co, 133Ba, 137Cs and 152Eu, standardized at the Nuclear Metrology Laboratory (NML) of IPEN. For lower activity of the impurities, the distance source-detector of 1.7 cm was assumed. However, as at this distance, the sum coincidence effect is very high, making the measurement of the standard calibration ampoules difficult, the spectrometer efficiency curve was obtained by a Monte Carlo simulation code, developed at IPEN. In this code, all details of the detection system are modeled and the response curves for x-rays and gamma rays are calculated by the MCNPX radiation transport code. The gamma spectra were analyzed by Alpino code, which applies the method of numeric peak integration of the area under the photopeaks. For gamma emitter impurities, not visually detected, the decision threshold and the detection limits were calculated from the background count rate, under the peak area. The radiopharmaceutical solutions analyzed were 67Ga,99Mo, 99mTc, 111In, 131I, 153Sm, 177Lu and 201Tl. The results of impurities ratio for analyzed solutions are in accordance with the manufacturers´ certificate and with the ANVISA.

Os radiofármacos são fármacos que possuem em sua composição um radionuclídeo e são utilizados para diagnóstico e tratamento de diversas doenças na medicina nuclear. O principal objetivo desse trabalho foi avaliar o efeito da temperatura e umidade na estabilidade de alguns reagentes liofilizados ALBUMINA-TEC, CIS-TEC, DEX-500-TEC, DEX-70-TEC, DISI-TEC, DMSA-TEC, DTPA-TEC, FITA-TEC, PUL-TEC, MIBI-TEC, MDP-TEC, PIRO-TEC, e TIN-TEC. Foi utilizada uma câmara climática para expor amostras destes produtos a uma temperatura de 40 °C e 75% de umidade relativa por 48 horas, para simular uma condição de transporte. Foram avaliadas duas situações: após um determinado tempo de armazenamento em temperatura de 2 - 8 °C, amostras de RL foram expostas por 48 horas em 40 °C e 75% UR e em seguida foram analisadas enquanto outras amostras foram colocadas por 48 horas em 40 °C e 75% UR, mantidas em temperatura de 2 - 8 °C até a validade do radiofármaco e analisadas quanto à pureza radioquímica e biodistribuição. Os radiofármacos ALBUMINA-TEC, DEX-70-TEC, DEX-500-TEC, DISI-TEC, DMSA-TEC, DTPA-TEC, PIRO-TEC, MDP-TEC, MIBI-TEC e MDP-TEC apresentaram resultados de % PRq e biodistribuição que atenderam às especificações, enquanto que CIS-TEC, PUL-TEC e TIN-TEC apresentaram alguns valores de % PRq menor que o especificado. TIN-TEC apresentou % PRq de 84% em um de três lotes, enquanto PUL-TEC apresentou % PRq de 88% em um de quatro lotes analisados; CIS-TEC é um RL que ainda precisa melhor analisado pois os resultados fora do especificado no controle radioquímico e biológico foram obtidos com um produto fora da validade especificada. Estes resultados indicaram que a pureza radioquímica pode ser afetada pela condição de estudo, o que não foi confirmado pelo controle biológico. / Radiopharmaceuticals are pharmaceuticals that have in their composition a radionuclide and are used for diagnosis and treatment of various diseases in nuclear medicine. The main objective of this study was to evaluate the effect of temperature and humidity on the stability of some lyophilized reagents: ALBUMINA-TEC, CIS-TEC, DEX-500-TEC, DEX-70-TEC, DISI-TEC, DMSA-TEC, DTPA-TEC, FITA-TEC, PUL-TEC, MIBI-TEC, MDP-TEC, PIRO-TEC and TIN-TEC. A climatic chamber was used for exposing some samples of these products to a temperature of 40 °C and 75% RH for 48 hours to simulate a transport condition. Two situations were evaluated: after a certain storage time at 2 - 8 °C, LR samples were exposed for 48 hours at 40 °C and 75% RH and analyzed while other samples were placed for 48 hours at 40 °C and 75% RH, kept at 2 - 8 °C until the expiration time of each radiopharmaceutical and analyzed for radiochemical purity and biodistribution. ALBUMINA-TEC, DEX-70-TEC, DEX-500-TEC, DISI-TEC, DMSA-TEC, DTPA-TEC, PIRO-TEC, MDP-TEC, MIBI-TEC and MDP-TEC presented results of % RCP and biodistribution that met the specifications while CIS-TEC PUL-TEC and TIN-TEC presented some % RCP lower than specified. TIN-TEC presented % RCP of 84% in one of three lots while PUL-TEC presented % RCP of 88% in a one of four batches analyzed; CIS-TEC is a LR that further need to be analyzed because probable the out of specification results of radiochemical and biological controls were obtained with a product outside of the validity date. These results indicated that radiochemical purity, can be affected by study condition, which has not been confirmed by biological control.

O radiofármaco FLT-18F é um análogo do nucleosídeo timidina e um promissor marcador da proliferação tumoral para imagens em PET. A síntese deste radiofármaco não é simples e, muitas vezes, apresenta baixos rendimentos. Este radiofármaco já vem sendo estudado há alguns anos, porém, não há produção, nem estudos clínicos, no Brasil. O estudo do processo produtivo e a sua adequação às diretrizes de Boas Práticas de Fabricação (ANVISA) são de extrema importância. Este trabalho teve como objetivo estudar a síntese deste radiofármaco, avaliar os métodos de controle de qualidade que serão utilizados na rotina de produção futura, realizar estudos de citotoxicidade, estudos de biodistribuição e imagens PET em animais, contribuindo para o desenvolvimento e elaboração do protocolo de validação de processo e estabelecimento das metodologias analíticas a serem utilizadas durante a rotina de produção. Inicialmente, foi estudada a síntese e produção do produto FLT-18F, com a avaliação de três temperaturas diferentes de marcação, a m de vericar o comportamento do rendimento radioquímico e a estabilidade do produto nal. Os estudos de metodologia analítica compreenderam as análises de identicação radionuclídica, determinação dos pers cromatográcos, pureza radioquímica, solventes residuais e pH. Estudos in vitro do FLT- 18F de internalização e citotoxicidade também foram feitos. Nos estudos in vivo, avaliou-se a farmacocinética, biodistribuição em animais sadios e em animais com modelos tumorais, além de imagens PET/CT de animais com melanoma. O produto nal apresentou alta pureza radioquímica e mostrou-se estável por até 10 horas após a síntese, porém obteve-se um rendimento radioquímico relativamente baixo, conforme descrito na literatura. As metodologias analíticas testadas mostraram-se adequadas para o uso no controle de qualidade do FLT-18F. Nos estudos in vitro o FLT-18F apresentou uma signicativa porcentagem de ligação às células tumorais e a molécula não radiomarcada não foi considerada tóxica para estas células estudadas. A biodistribuição e as imagens apresentaram resultados compatíveis com o esperado. As contribuições para a validação de processo foram satisfatórias e auxiliarão na validação futura do processo produtivo do radiofármaco em estudo. / Radiopharmaceutical 18F-FLT is a thymidine nucleoside analogue and a promising tumor proliferation marker for PET images. The synthesis of this radiopharmaceutical is not simple, and often has low yields. This radiopharmaceutical has already been studied for some years; however, there is no production, nor are there clinical studies in Brazil. The study of the production process and its compliance with the guidelines of Good Manufacturing Practices (ANVISA) are of extreme importance. This study aimed to investigate the synthesis of this radiopharmaceutical, evaluate methods of quality control that will be used in future production routines, perform cytotoxicity studies, biodistribution studies and PET imaging in animals, thereby contributing to the development and elaboration of the process validation protocol and to the establishment of analytical methods to be used during production routines. Initially, we studied the synthesis and production of 18F-FLT, with the evaluation of three dierent temperatures of radiolabeling to check the behavior of the radiochemical yield and stability of the nal product. Studies of analytical methodology comprised the analysis of radionuclide identication, determination of chromatographic proles, radiochemical purity, residual solvents, and pH. In vitro studies of internalization and cytotoxicity were also carried out. In in vivo studies, we evaluated the pharmacokinetics, biodistribution in healthy animals and in animals with tumor models, in addition to PET/CT images in animals with melanomas. The nal product had high radiochemical purity and was stable for up to 10 hours after the synthesis, but got a relatively low radiochemical yield, as described in the literature. The tested analytical methods proved suitable for use in the quality control of 18F-FLT. In in vitro studies, 18F-FLT showed a signicant percentage of binding to tumor cells, and the nonradiolabeled molecule was not considered toxic for these studied cells. The biodistribution and images showed results that were consistent with expectations. Contributions to the validation process were satisfactory, and will assist in the future validation of the production process of the radiotracer under study.

O radiofármaco FLT-18F é um análogo do nucleosídeo timidina e um promissor marcador da proliferação tumoral para imagens em PET. A síntese deste radiofármaco não é simples e, muitas vezes, apresenta baixos rendimentos. Este radiofármaco já vem sendo estudado há alguns anos, porém, não há produção, nem estudos clínicos, no Brasil. O estudo do processo produtivo e a sua adequação às diretrizes de Boas Práticas de Fabricação (ANVISA) são de extrema importância. Este trabalho teve como objetivo estudar a síntese deste radiofármaco, avaliar os métodos de controle de qualidade que serão utilizados na rotina de produção futura, realizar estudos de citotoxicidade, estudos de biodistribuição e imagens PET em animais, contribuindo para o desenvolvimento e elaboração do protocolo de validação de processo e estabelecimento das metodologias analíticas a serem utilizadas durante a rotina de produção. Inicialmente, foi estudada a síntese e produção do produto FLT-18F, com a avaliação de três temperaturas diferentes de marcação, a m de vericar o comportamento do rendimento radioquímico e a estabilidade do produto nal. Os estudos de metodologia analítica compreenderam as análises de identicação radionuclídica, determinação dos pers cromatográcos, pureza radioquímica, solventes residuais e pH. Estudos in vitro do FLT- 18F de internalização e citotoxicidade também foram feitos. Nos estudos in vivo, avaliou-se a farmacocinética, biodistribuição em animais sadios e em animais com modelos tumorais, além de imagens PET/CT de animais com melanoma. O produto nal apresentou alta pureza radioquímica e mostrou-se estável por até 10 horas após a síntese, porém obteve-se um rendimento radioquímico relativamente baixo, conforme descrito na literatura. As metodologias analíticas testadas mostraram-se adequadas para o uso no controle de qualidade do FLT-18F. Nos estudos in vitro o FLT-18F apresentou uma signicativa porcentagem de ligação às células tumorais e a molécula não radiomarcada não foi considerada tóxica para estas células estudadas. A biodistribuição e as imagens apresentaram resultados compatíveis com o esperado. As contribuições para a validação de processo foram satisfatórias e auxiliarão na validação futura do processo produtivo do radiofármaco em estudo. / Radiopharmaceutical 18F-FLT is a thymidine nucleoside analogue and a promising tumor proliferation marker for PET images. The synthesis of this radiopharmaceutical is not simple, and often has low yields. This radiopharmaceutical has already been studied for some years; however, there is no production, nor are there clinical studies in Brazil. The study of the production process and its compliance with the guidelines of Good Manufacturing Practices (ANVISA) are of extreme importance. This study aimed to investigate the synthesis of this radiopharmaceutical, evaluate methods of quality control that will be used in future production routines, perform cytotoxicity studies, biodistribution studies and PET imaging in animals, thereby contributing to the development and elaboration of the process validation protocol and to the establishment of analytical methods to be used during production routines. Initially, we studied the synthesis and production of 18F-FLT, with the evaluation of three dierent temperatures of radiolabeling to check the behavior of the radiochemical yield and stability of the nal product. Studies of analytical methodology comprised the analysis of radionuclide identication, determination of chromatographic proles, radiochemical purity, residual solvents, and pH. In vitro studies of internalization and cytotoxicity were also carried out. In in vivo studies, we evaluated the pharmacokinetics, biodistribution in healthy animals and in animals with tumor models, in addition to PET/CT images in animals with melanomas. The nal product had high radiochemical purity and was stable for up to 10 hours after the synthesis, but got a relatively low radiochemical yield, as described in the literature. The tested analytical methods proved suitable for use in the quality control of 18F-FLT. In in vitro studies, 18F-FLT showed a signicant percentage of binding to tumor cells, and the nonradiolabeled molecule was not considered toxic for these studied cells. The biodistribution and images showed results that were consistent with expectations. Contributions to the validation process were satisfactory, and will assist in the future validation of the production process of the radiotracer under study.

Este trabalho tem como objetivo investigar a concentração de impurezas radioativas gama emissoras presentes nas soluções dos radiofármacos produzidos no Instituto de Pesquisas Energéticas e Nucleares - IPEN em São Paulo. Para que este radiofármaco possa ser utilizado adequadamente, sua qualidade deve ser avaliada de acordo com os procedimentos estabelecidos de acordo com os \"Requisitos Gerais para a Competência de Laboratórios de Teste e Calibração\", ISO / IEC 17025: 2005 e pelas \"Boas Práticas de Fabricação\" (BPF), controladas pela ANVISA (Agência Nacional de Vigilância Sanitária), no Brasil. Para determinar a atividade, dos radiofármacos das impurezas gama emissoras, foi utilizado um espectrômetro gama de alta resolução em duas distâncias fonte-detector; uma de 18 cm e outra de 1,7 cm. Para a distância de 18 cm, o espectrômetro HPGe foi calibrado com energias gama entre 81 kev e 1408 kev, medindo ampolas seladas de 60Co, 133Ba, 137Cs e 152Eu, padronizadas no Laboratório de Metrologia Nuclear (LMN) do IPEN. Para impurezas com baixas atividades, utilizou a distância fontedetector de 1,7 cm. A esta distância, o efeito soma em cascata é muito elevado, tornando difícil a medição das ampolas de calibração padrão, com isso, a curva de eficiência do espectrômetro foi obtida por um código de simulação de Monte Carlo, desenvolvido no IPEN. Neste código, todos os detalhes do sistema de detecção são modelados e as curvas de resposta para raios X e raios gama são calculadas pelo código de transporte de radiação MCNPX. Os espectros gama foram analisados pelo programa Alpino, que aplica o método de integração numérica da área sob os fotopicos de absorção total. Para as impurezas gama emissoras não detectadas visualmente, os limites de detecção foram calculados a partir da taxa de contagem de fundo, sob a área do pico de interesse. As soluções radioativas analisadas foram 67Ga,99Mo, 99mTc, 111In, 131I, 153Sm, 177Lu e 201Tl. Os resultados da relação entre a atividade do radionuclídeo em análise e as impurezas identificadas apresentaram acordo com os certificados de análise dos fabricantes, assim como, com as especificações da ANVISA. / This work aims to investigate the concentration of radioactive impurities gamma emitters in the radiopharmaceutical solutions produced at Nuclear and Energy Research Institute -IPEN in São Paulo, So that this radiopharmaceutical may be used properly, its quality should be evaluated in accordance with the procedures established by quality control agencies, such as \"General Requirements for the Competence of Testing and Calibration Laboratories\", ISO/IEC 17025:2005 and the \"Good Laboratory Practice\" (GLP), controlled by ANVISA (National Agency Health Surveillance), in Brazil, requiring a confirmation of the values of impurities related at the certificates supplied by the manufacturers. To determine the activity, a high resolution gamma spectrometer were used in two source-detector distances. One was 18 cm and the other 1.7 cm. For the 18 cm distance, the high pure germanium spectrometer was calibrated in the energy range between 81 keV and 1408 keV by measuring sealed ampoules of 60Co, 133Ba, 137Cs and 152Eu, standardized at the Nuclear Metrology Laboratory (NML) of IPEN. For lower activity of the impurities, the distance source-detector of 1.7 cm was assumed. However, as at this distance, the sum coincidence effect is very high, making the measurement of the standard calibration ampoules difficult, the spectrometer efficiency curve was obtained by a Monte Carlo simulation code, developed at IPEN. In this code, all details of the detection system are modeled and the response curves for x-rays and gamma rays are calculated by the MCNPX radiation transport code. The gamma spectra were analyzed by Alpino code, which applies the method of numeric peak integration of the area under the photopeaks. For gamma emitter impurities, not visually detected, the decision threshold and the detection limits were calculated from the background count rate, under the peak area. The radiopharmaceutical solutions analyzed were 67Ga,99Mo, 99mTc, 111In, 131I, 153Sm, 177Lu and 201Tl. The results of impurities ratio for analyzed solutions are in accordance with the manufacturers´ certificate and with the ANVISA.