A combined experimental and mathematical approach for molecular-based optimization of irinotecan circadian delivery

Circadian timing largely modifies efficacy and toxicity of many anticancer drugs. Recent findings suggest that optimal circadian delivery patterns depend on the patient genetic background. We present here a combined experimental and mathematical approach for the design of chronomodulated administration schedules tailored to the patient molecular profile. As a proof of concept we optimized exposure of Caco-2 colon cancer cells to irinotecan (CPT11), a cytotoxic drug approved for the treatment of colorectal cancer. CPT11 was bioactivated into SN38 and its efflux was mediated by ATP-Binding-Cassette (ABC) transporters in Caco-2 cells. After cell synchronization with a serum shock defining Circadian Time (CT) 0, circadian rhythms with a period of 26 h 50 (SD 63 min) were observed in the mRNA expression of clock genes REV-ERBα, PER2, BMAL1, the drug target topoisomerase 1 (TOP1), the activation enzyme carboxylesterase 2 (CES2), the deactivation enzyme UDP-glucuronosyltransferase 1, polypeptide A1 (UGT1A1), and efflux transporters ABCB1, ABCC1, ABCC2 and ABCG2. DNA-bound TOP1 protein amount in presence of CPT11, a marker of the drug PD, also displayed circadian variations. A mathematical model of CPT11 molecular pharmacokinetics-pharmacodynamics (PK-PD) was designed and fitted to experimental data. It predicted that CPT11 bioactivation was the main determinant of CPT11 PD circadian rhythm. We then adopted the therapeutics strategy of maximizing efficacy in non-synchronized cells, considered as cancer cells, under a constraint of maximum toxicity in synchronized cells, representing healthy ones. We considered exposure schemes in the form of an initial concentration of CPT11 given at a particular CT, over a duration ranging from 1 to 27 h. For any dose of CPT11, optimal exposure durations varied from 3h40 to 7h10. Optimal schemes started between CT2h10 and CT2h30, a time interval corresponding to 1h30 to 1h50 before the nadir of CPT11 bioactivation rhythm in healthy cells

Amine Functionalization of Bacterial Cellulose for Targeted Delivery Applications

Bacterial cellulose (BC), produced by acetic acid bacteria Gluconacetobacter xylinus, is ideal for delivery and related biomedical functions. It is FDA approved for wound dressings and internal applications, non-toxic to endothelial cells and has little effect on blood profiles. Conjugation of therapeutics to BC can be accomplished through the available alcohol groups of the anhydroglucose units (AGU), making targeted delivery possible. Amine was introduced to BC through a reaction involving epichlorohydrin and ammonium hydroxide. The chemical structure was analyzed using infrared spectroscopy and quantified through pH titration. Conjugation of amine to BC was demonstrated through fluorescein-5’-isothiocyanate (FITC) and bromocresol green (BCG) attachment. Due the its large molecular size, the protein horseradish peroxidise (HRP) was conjugated to aminated-BC through a bis(sulfosuccinimidyl)suberate (BS3) linker to reduce steric congestion on the BC surface. Hydrogen peroxide was used to hydrolyze BC to create nanocrystalline cellulose (NCC-BC) with dimensions capable of intracellular delivery. Amine was introduced to NCC-BC and the chemical structure was analyzed using infrared spectroscopy and quantified through pH titration. HRP was optimized to demonstrate protein attachment, while avidin-HRP was used to demonstrate the ability of maximizing protein loading. An avidin-biotin glucose oxidase and avidin-biotin β-galactosidase complex was conjugated to aminated NCC-BC to demonstrate the application of a drug carrier of therapeutic proteins

Innovation in diabetic care : from patient-centered care to public policies to reduce the impact of diabetes

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2022Introdução A diabetes, nomeadamente a diabetes tipo 2, é uma doença crónica, que necessita de cuidados de saúde em continuidade, de longa duração e muitas vezes multidisciplinares. A diabetes consome vastos recursos de saúde, financeiros e de assistência social, exigindo, em todos os níveis de cuidados de saúde, e às famílias um grande esforço. Sendo uma doença que exige capacitação do paciente, literacia, adesão ao tratamento e promoção de comportamentos saudáveis, é também o paradigma de uma doença onde a relação com os profissionais de saúde e o cuidado centrado no paciente são fatores fundamentais para seu controle. A saúde tem sido palco de muitas inovações, tanto tecnológicas, como também de gestão, prática clínica, farmacológicas, dispositivos vários e uso da informação. Os cuidados na diabetes e outras doenças crónicas estão na linha de frente do desenvolvimento, testagem e implementação destas inovações, o que iremos ilustrar ao longo deste trabalho. Objetivo Identificar e analisar como a inovação nos cuidados prestados aos doentes com diabetes tem vindo a alterar a capacidade de prestar cuidados centrados no doente, e demonstrar abertura a novas iniciativas e ao desenvolvimento de políticas públicas de gestão em saúde, financiamento, tecnologia/informação e sociais com vista providenciar melhores cuidados e reduzir os impactos desta doença. Neste contexto, é nosso objetivo descrever, conceptualizar e sistematizar como a inovação tem evoluído e influenciado os cuidados ao paciente diabético e identificar os respetivos impactos. Métodos Procedeu-se à revisão da literatura usando as bases de dados PubMed, Scopus e Word of Science, pesquisando a associação entre diabetes tipo 2 e inovação. Foram encontrados 254 artigos. A partir da seleção desses trabalhos foi efetuada uma busca manual de artigos a descrever, ilustrar e avaliar as inovações identificadas, num total de 69 artigos.Resultados No âmbito dos cuidados aos doentes com diabetes, foram identificados dezasseis processos de inovação com inegável relevância em quatro áreas – três casos de inovação em gestão, quatro financeiras, seis tecnológicas e três de ação social. Conclusão Constata-se que as diversas categorias de inovação estão interligadas e são complementares, possibilitando oferecer melhores cuidados centrados no paciente, ao mesmo tempo que vislumbram a necessidade de proceder ao “redesenho” dos sistemas e serviços de saúde. Admite-se que, no futuro, a sua adequada integração acarretaria melhores cuidados e permitiria reduzir o impacto da diabetes, podendo vir a ser o modelo a utilizar no manejo de outras doenças crónicas.Background Diabetes, namely type-2 diabetes, is a chronic disease that requires continuous, long-term, and often multidisciplinary medical care. Diabetes consumes a vast amount of health, financial and social care resources, demanding great efforts at all levels of health care and families. As a disease that requires patient training, literacy, adherence to treatment, and the promotion of healthy behaviors, it is also the paradigm of a disease where health professionals’ relationship and patient-centered care are key factors for its control. Health has been the scene of many innovations, both technological, as well as management, clinical practice, medication, a wide variety of devices, and information uses. Diabetic, and of other chronic diseases, is at the forefront of the development, testing, and implementation of these innovations, which we will illustrate throughout this work. Aim Identify and analyze how innovation in care provided to patients with diabetes has been changing the ability to provide patient-centered care, and open-up new initiatives and development of public policies in health, financing, technological/information and social management with the view to provide better care and reduce the impacts of this disease. In this context, we will describe, conceptualize and systematize how innovation has evolved and influenced diabetic patient care and identify the respective impacts. Methods A literature review was carried out using PubMed, Scopus, and Word of Science databases, investigating the association between type 2 diabetes and innovation. Two hundred and fifty-four articles were found. From the research, a manual search of articles aiming to describe, exemplify, and evaluate the innovations in diabetes care, was carried out, resulting in a total of 69 articles.Results Four main areas were identified within the scope of diabetes patients’ care, in which innovation processes are present with undeniable relevance – management, financial, technological, and social action, which will be analyzed on the course of this work. Conclusion It was possible to observe that the different categories of innovation are interconnected and complementary, making it possible to offer better patient-centered care, while at the same time envisioning the need to “redesign” of health systems and services. It is accepted that, in the future, their appropriate integration would lead to better care and reduce the impact of diabetes, as well as potentially become the model that could be used in the management of other chronic diseases

non-small cell lung cancer

Trabalho Final de Mestrado Integrado, Ciências Farmacêuticas, 2022, Universidade de Lisboa, Faculdade de Farmácia.O cancro é um problema mundial e, apesar dos avanços da ciência e medicina na compreensão da sua biologia e genética, é a segunda causa de morte. Durante muito tempo, o seu tratamento baseou-se na cirurgia, radioterapia e quimioterapia. Embora estas terapias tenham apresentado resultados positivos, apresenta taxas de sucesso variáveis devido à falta de especificidade e toxicidade sistémica inerente. A medicina de precisão introduz uma mudança de paradigma no campo médico, mostrando-se promissora no diagnóstico atempado, bem como na melhoria das condições de vida dos doentes com cancro. O objetivo é avaliar dados genéticos, estilo de vida e ambientais do paciente, com recurso a tecnologia de ponta, como testes moleculares e sequenciamento de última geração, para fornecer um diagnóstico preciso num estadio inicial, bem como um tratamento apropriado. Assim, a medicina de precisão pode permitir a prevenção de doenças e, ao mesmo tempo, reduzir os custos e efeitos colaterais da terapêutica. Atualmente, o campo com maior pesquisa e aplicação em medicina de precisão, é a oncologia. O cancro do pulmão é um exemplo proeminente do sucesso desta inovação no tratamento de tumores sólidos malignos. O cancro de pulmão é um grande problema de saúde pública, sendo a principal causa mundial de morte relacionada com o cancro. Isto deve-se a um diagnóstico em estadios avançados, em consequência da ausência de sintomas na fase inicial da doença, o que a torna muitas vezes incurável. Assim, a realização do perfil molecular é uma ferramenta ideal para o diagnóstico e deteção atempados. A caracterização molecular abrangente do cancro de pulmão expandiu a nossa perceção das origens celulares e vias moleculares afetadas em cada um dos subtipos. Além disso, muitas das alterações genéticas encontradas representam potenciais alvos terapêuticos (biomarcadores) para os quais novos fármacos estão em constante desenvolvimento. Esta monografia pretende dar uma visão geral da atual medicina de precisão no cancro, bem como as suas perspetivas futuras. Foi dado destaque ao cancro do pulmão, mais precisamente ao subtipo de cancro do pulmão de células não pequenas, devido à sua alta incidência e mortalidade.Cancer is a worldwide problem, and despite tremendous advances in science and medicine in the understanding of its biology and genetics, it is still a major cause of death. For a long time, surgery, radiotherapy, and chemotherapy were the most used approaches for cancer treatment. Even though these therapies have shown positive outcomes, variable success rates have been obtained due to their lack of specificity and inherent systemic toxicity. Precision medicine is a new, paradigm-shift approach in medical field that shows promise for improving many aspects of healthcare, both in terms of diagnosis and treatment of diseases. The goal of precision medicine is to use cutting-edge healthcare resources, such as molecular tests and next-generation sequencing to decipher patient's genomic, lifestyle, and environmental data, and thereby identify a precise diagnosis at an earlier stage, as wells as an appropriate treatment. In this way, precision medicine may allow the prevention of disease while also reducing the costs and side effects of therapy. Currently, oncology is the field with major research and application of precision medicine. A prominent example of the success of this innovation in treating solid tumour malignancies, is lung cancer. Lung cancer is a major public health concern, being the leading cause of cancer related mortality worldwide. The reason for this is the diagnosis at an advance stage, due to the absence of symptoms at early stages, which makes the disease often incurable. As such, molecular profiling isthe ideal tool for early diagnosis and detection. Comprehensive molecular characterization of lung cancer has expanded our understanding of the cellular origins and molecular pathways affected in each of the sub-types. Furthermore, many of these genetic alterations represent potential therapeutic targets (biomarkers) for which novel drugs are constantly under development. This monograph aims to give an overview of cancer precision medicine current state and future perspectives. A special focus was given to lung cancer, more precisely to the non-small cell lung cancer subtype, due to its high incidence and mortality