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N an O Te Ch N O Lo Gy-B Ase D S Trate Gie S Fo R R E Ctal a N Ti-H IV P Re Rute Sofia Nunes Rute Sofia HIV Pre Nanotechnology Anti Rectal Rute Sofia Nunes Rute Sofia Nunes Rute Sofia HIV Pre Nanotechnology Rectal Anti Rute Sofia Nunes Rute Sofia Nunes Rute Sofia Nanotechnology Rute Sofia Nunes Sofia Nunes DOUTORAMENTO CIÊNCIAS BIOMÉDICAS INSTITUTO DE CIÊNCIAS BIOMÉDICAS ABEL SALAZAR ABEL BIOMÉDICAS CIÊNCIAS DE INSTITUTO INSTITUTO DE CIÊNCIAS BIOMÉDICAS ABEL SALAZAR ABEL BIOMÉDICAS CIÊNCIAS DE INSTITUTO INSTITUTO DE CIÊNCIAS BIOMÉDICAS ABEL SALAZAR ABEL BIOMÉDICAS CIÊNCIAS DE INSTITUTO INSTITUTO DE CIÊNCIAS BIOMÉDICAS ABELSALAZAR ABELSALAZAR BIOMÉDICAS BIOMÉDICAS CIÊNCIAS DE CIÊNCIAS DE INSTITUTO INSTITUTO DE CIÊNCIAS BIOMÉDICAS ABEL SALAZAR BIOMÉDICAS CIÊNCIAS DE INSTITUTO INSTITUTO DE CIÊNCIAS BIOMÉDICAS ABEL SALAZAR ABEL BIOMÉDICAS CIÊNCIAS DE INSTITUTO INSTITUTO DE CIÊNCIAS BIOMÉDICAS ABEL SALAZAR ABEL BIOMÉDICAS CIÊNCIAS DE INSTITUTO Nanotechnology- Based Strategies for - Exposure Prophylaxis Exposure Prophylaxis Exposure Prophylaxis Exposure Prophylaxis Exposure Exposure Prophylaxis Exposure - HIV Pre HIV HIV Pre HIV Pre HIV Rectal Anti- HIV Pre- Exposure - . Based Strategies for Rectal A Based Strategies Strategies for for Rectal Rectal A A Based Strategies for Rectal A Based Strategies for Rectal A Based Based Strategies for Rectal A - Exposure Prophylaxis ExposureProphylaxis ExposureProphylaxis ExposureProphylaxis Prophylaxis Nanotechnology Nanotechnology Rute Sofia Nunes - - Based Strategies for Based Strategies Strategies for for Based Strategies for nti nti - D .ICBAS .ICBAS 2018 2018 Rute Sofia Gonçalves Nunes Nanotechnology - Based Strategies for Rectal Anti - HIV Pre - Exposure Prophylaxis Tese de Candidatura ao grau de Doutor em Ciências Biomédicas submetida ao Instituto de Ciências Biomédicas Abel Salazar da Universidade do Porto. Orientador Doutor Bruno Filipe Carmelino Cardoso Sarmento Categoria – Investigador Auxiliar/Professor Auxiliar Afiliação – i3S - Instituto de Investigação e Inovação em Saúde, INEB - Instituto de Engenharia Biomédica, Universidade do Porto & IUCS - Instituto Universitário de Ciências da Saúde Co - orientador Doutor José Alberto Gonçalves das Neves Categoria – Inves tigador Pós - Doutorado Afiliação – i3S - Instituto de Investigação e Inovação em Saúde & INEB - Instituto de Engenharia Biomédica Tutor Doutor Pedro Lopes Granja Categoria – Investigador Principal/Professor Associado Afiliação – i3S - Instituto de Investigação e Inovação em Saúde, INEB - Instituto de Engenharia Biomédica & Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto The work presented in this thesis was developed at: Nanomedicines & Translational Drug Delivery i3S - Instituto de Investigação e Inovação em Saúde and INEB - Instituto Nacional de Engenharia Biomédica Universidade do Porto, Porto, Portugal Rua Alfredo Allen, 208 4200 - 135 Porto, Portugal www.i3s.up.pt | www.ineb.up.pt Financial Support Rute Nunes was supported by a national PhD grant (SFRH/BD/96519/2013) from Fundação para a Ciência e Tecnologia (FCT), Portugal. This work is a result of the project NORTE - 01 - 0145 - FEDER - 000012, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). This work was finance d by FEDER - Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020 - Operacional Programme for Competitiveness and Internationalisation (POCI), Portugal 2020, and by Portuguese funds through FCT - Fundação para a Ciência e a Tecnologia/Mi nistério da Ciência, Tecnologia e Ensino Superior in the framework of the project "Institute for Research and Innovation in Health Sciences" (POCI - 01 - 0145 - FEDER - 007274), and also Partnership Agreement PT2020 UID/QUI/50006/2013 - POCI/01/0145/FEDER/007265. Gilead Sciences, Lda., Portugal is also acknowledged for support under the framework of the Programa Gilead GÉNESE (ref. PGG/002/2016). Publications Ao abrigo do disposto do n.º 2, alínea a) do artigo 31.º do Decreto - Lei n.º 115/2013 de 7 de Agosto, fazem parte integrante desta tese de doutoramento os seguintes trabalhos já publicados ou submetidos para publicação: Melo M*, Nunes R* , das Neves J, Sarmento B. Rectal administration of nanosystems: from drug delivery to diagnostics. Submitted for publication . * Equal contribution Nunes R , Araújo F, Barreiros L, Bártolo I, Segundo MA, Taveira N, et al. Non - covalent PEG coating of nanoparticle drug carriers improves the local pharmacokinetics of rectal anti - HIV microbicides. Submitted for publication . Nunes R , Araújo F, Tavares J, Sarmento B, das Neves J. Surface modification with polyethylene glycol en hances colorectal distribution and retention of nanoparticles. Eur J Pharm Biopharm. 2018;130:200 - 6. Nunes R , Silva C, Chaves L. Tissue - based in vitro and ex vivo models for intestinal permeability studies. In: Sarmento B, editor. Concepts and Models for Drug Permeability Studies: Woodhead Publishing; 2016. p. 203 - 36. Nunes R , Sarmento B, das Neves J. Formulation and delivery of anti - HIV rectal microbicides: Advances and challenges. J Control Release. 2014;194c:278 - 94. Nunes R , Sousa C, Sarmento B, das N eves J. Nanotechnology - based systems for microbicide development. In: das Neves J, Sarmento B, editors. Drug Delivery and Development of Anti - HIV Microbicides. Singapore: Pan Stanford; 2014. p. 415 - 58. Acknowledgments “Aqueles que passam por nós, não vão sós, não nos deixam sós. Deixam um pouco de si, levam um pouco de nós.” Antoine de Saint - Exupéry Foram várias as pessoas que ao longo deste percurso, directa ou indirectamente, ajudaram na concretização deste trabalho. Deixo aqui o meu agradecimento e gratidão a todas elas. No entanto, não poderia deixar de dedicar um agradecimento especial às seguintes pessoas: Ao Professor Doutor Bruno Sarmento, o meu Orientador, pela oportunidade de fazer parte do seu grupo de Investigação, por estar sempre presente mesmo quando fisicamente ausente, pelo esforço e dedicação diários para garantir o suporte das nossas actividades científicas. Agradeço a sua compreensão, a sua paciência, a sua objectividade na abordagem das questões e a sua orientação ci entífica. Ao Doutor José das Neves, o meu Co - orientador, a quem um Muito Obrigada nunca será suficiente para agradecer toda a dedicação, empenho e paciência que teve durante todo este percurso. O pensamento crítico, o questionar constante (mais do que obt er respostas), a busca incessante pelo conhecimento e a discussão de ideias serão inequivocamente uma herança da sua orientação. Obrigada por todos os ensinamentos, por toda a ajuda, por todos os sensatos conselhos científicos, por toda a confiança deposit ada em mim e no meu trabalho. Consigo aprendi que suficiente não satisfaz, bom não é suficiente e muito bom tem de ser perfeito. “Se não for para fazer bem, é melhor não fazer.” Certamente, graças à sua orientação, cresci enquanto investigadora. Ao Profe ssor Doutor Pedro Granja, o meu Tutor, pela oportunidade de fazer parte integrante do INEB. Ao Professor Doutor Nuno Taveira, pela sua ajuda, pela disponibilidade e entusiasmo com que me recebeu no seu laboratório. A toda a sua equipa de investigação pelo bem receber e acolher e pela constante animação. Um agradecimento especial à Doutora Inês Bártolo, por toda a sua ajuda, apoio e disponibilidade na realização dos ensaios de actividade anti - retroviral. Ao Professor Vítor Seabra, pel a sua inteira disponibilidade e por ter proporcionado todas as condições para a realização de ensaios in vivo na CESPU, Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da Saúde. À Professora Doutora Marcela Segundo, da Faculdade d e Farmácia da Universidade do Porto, por prontamente ter aceitado fazer no seu laboratório os doseamentos do fármaco por HPLC - MS/MS. Obrigada a toda a sua equipa de investigação por me fazer sentir em casa. Um agradecimento especial à Doutora Luísa Barreir os, por todo o tempo e ajuda dedicados a esta parte do trabalho. À Dr. Virgínia Gonçalves, do IINFACTS, CESPU, pela amizade, e por todo o apoio prestado nas experiências de cromatografia líquida de alta eficiência, bem como em questões de logística de laboratório. Ao Doutor Carlos Fernandes, da Faculdade de Ciências da Universidade do Porto, pela ajuda imprescindível nas experiências de ressonância magnética nuclear de protões. À Doutora Fabíola Moutinho, do INEB, pela ajuda e prontidão nas experiências de medição de ângulo de contacto. A todos os membros da “BS Team” sem excepção, os que passaram e os que ainda cá estão, o meu Muito Obrigada pela troca de experiências, pela ajuda, pelos bons momentos e sorrisos. Convosco a viagem tornou - se mais fácil! O meu agradecimento especial à minha companheira de viagem, Ana Costa, por todos os momentos partilhados, por todos os sorrisos cúmplices, por todas as confidências, por todas as gargalhadas escancaradas nos momentos menos oportunos, por todos os desabafos nos momentos de frustração e desânimo, por todos os pôr - do - sol após mais um dia de trabalho sem resultados animadores. Ao Pedro Fonte, pela forma afável como nos acolheu no IINFACTS, CESPU, pela sua constante disponibilidade para ajudar, pela sua boa - disposição e pela partilha dos seus conhecimentos. À Cass ilda Reis, pela amizade, pela sua capacidade de bem ouvir e sensatez nos seus conselhos. Obrigada pela leveza que trouxeste a este percurso, por me mostrares que “o copo está sempre meio cheio”. Agradeço toda a ajuda e disponibilidade, quer pessoal
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