BRINGING PHARMACY KNOWLEDGE AND STUDENTS TOGETHER

Volume 3 | Edition 1 | October 2015 www.epsa-online.org EPSA Students' Science Publication

INTRODUCTION

Dear friends,

Another year has passed and it is time for the first edition of the EPSA Students' Science Publication (ESSP) of this academic year. The ESSP is a project that allows pharmacy students from around the Europe to publish their scientific research and outcomes. All of their abstracts are professionally reviewed by the European Federation for Pharmaceutical Sciences (EUFEPS). The project thus allows the students insight in the process of writing and publishing abstracts and, by extension, scientific articles. Every student also has the opportunity to present themselves and their motivation for conducting the research. Hopefully reading this edition inspires more students to firstly, try your hand at research and secondly, write and submit an abstract.

This edition contains four different abstract, covering various topics. You will learn more about 'Biomedical Potential of - (Bert.) Bertoni', 'Celebrating One Century and Thinking About the Future: Preliminary Findings for a New MPharm Syllabus at Lisbon Faculty of Pharmacy', 'Synthesis of a Novel 14-membered Oxo-thia-aza Macrocyclic Compound' and 'The Influence of Acute Pain on Osmotic Pressure and Cortisol Concentration in Tears'.

I would like to thank EUFEPS and the EPSA Public Relations Department for their hard work on this edition and all the students who collaborated on it. This would not have been possible without all of you. Looking forward to the future editions!

Eva Shannon Schiffrer, Science Coordinator 2015-2016

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TABLE OF CONTENT

Synthesis of a novel 14-membered oxo-thia-aza macrocyclic compound Abstract ...... 4 Q&A of the author ...... 5

Biomedical potential of Stevia - Stevia rebaudiana (Bert.) Bertoni Abstract ...... 6 Q&A of the author ...... 7

Celebrating one century and thinking about the future: preliminary findings for a new MPharm syllabus at Lisbon Faculty of Pharmacy Abstract ...... 8 Q&A of the author ...... 9

The influence of acute pain on osmotic pressure and cortisol concentration in tears Abstract ...... 10 Q&A of the author ...... 11

European Federation for Pharmaceutical Sciences (EUFEPS) Information ...... 12 Upcoming conferences ...... 13

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SYNTHESIS OF A NOVEL 14-MEMBERED OXO-THIA-AZA MACROCYCLIC COMPOUND João Franco Machado, Judite Costa, Maria F. Cabral Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Av. Professor Gama Pinto, 1649-003 Lisbon – Portugal

Background: Macrocyclic compounds have been targets of a widespread interest in the last decade due to their similarity to the countless analogue scaffolds found in a variety of biological systems. The unique structural, kinetic and thermodynamic features of macrocycles, as well as their favourable drug- like properties, have provided a motivation for further study and has had an enormous impact on the fields of medicine, pharmacy, chemistry and biology. However, this type of is yet under-exploited as new, effective drugs and research continues to be fairly limited.

Objective: Following our previous studies, we report the design, synthesis and structural characterization of a novel oxo-thia-aza macrocyclic compound, the dioxo-[14]aneN3S (1-thia-4,8,12-triazacyclotetradecane-3,13-dione).

Materials and Methods: Dioxo-[14]aneN3S was synthesised by reacting dimethyl thiodiglycolate, previously obtained by Fischer-Speier esterification technique (η = 89.6 %), with dipropylenetriamine in a large excess of dry methanol at 40 ºC, under nitrogen atmosphere for 9 days. This procedure was based on the Tabushi methodology for macrocyclization, which consists of the condensation of the dimethyl of an α,ω-dicarboxylic acid with a commercially available polyethylenepolyamine. The pure product was then structurally characterized by infrared and nuclear magnetic resonance (1H, 13C, COSY, NOESY, HSQC, HMBC) spectroscopies and also by mass spectrometry.

Results and Conclusion: The improvements in the reaction conditions introduced to the Tabushi general method (controlled temperature, N2 atmosphere, increased reaction time, and use of a dry solvent in a large excess) contributed to achieving - after purification by column chromatography - the desired macrocycle with a very high yield (η = 76.8 %), compared to the related compounds synthesised by Tabushi methodology reported in literature (η = 10

~ 20 %). The spectroscopic and spectrometric analyses of dioxo-[14]aneN3S allowed to confirm its chemical structure as well prove its high level of purity.

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Q&A OF THE AUTHOR

General information about João Franco Machado 22 years old, Sintra, Portugal, Faculty of Pharmacy, University of Lisbon, [email protected]

Why did you select this topic for your research? I am a trainee volunteer at the Research Institute for Medicines, where I have been developing a research work in the field of coordination chemistry of macrocyclic compounds through three years. During this time, I found out that macrocyclic ligands have a great potential to be developed as very promising therapeutic agents due to their unique physicochemical proprieties and also to their range on medical applications, which had motivated me to design and study novel and effective macrocycles.

How do you feel about having coped with the challenges that research brings? When I started the research project that I am now reporting, I already had a considerable amount of experience which turned out to facilitate my research work. However, each project is a completely new challenge, because when we are designing a novel macrocycle, it is possible to figure out that we can’t properly synthesise it or that it doesn´t show the proprieties we are expecting. But at the end, and despite all difficulties we can face, the feeling of getting a novel and promising pharmaceutical compound is very enthusiastic!

What skills did you develop as a researcher during your research period? During my research period I have not only improved my knowledge in the different fields of chemistry, but I also developed a set of valuable skills, such as patience, perseverance, dedication, hardworking, time management, adaptability, improvising, and critical thinking.

Would you like to share anything else with the students in Europe? Any recommendations? The most important advice I can give is to never give up, even if some of your objectives seem to be quite unlikely to accomplish. Remember that the dedication leads to the success and at the end, you will experience the most rewarding feeling you can’t even imagine!

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BIOMEDICAL POTENTIAL OF STEVIA - STEVIA REBAUDIANA (BERT.) BERTONI Ana Begić and Josip Barilar, Faculty of Pharmacy and Biochemistry, Zagreb, Croatia

Introduction: Stevia rebaudiana (Bert.) Bertoni (stevia) is a native plant to South America that is spread worldwide by cultivation due to its use as a natural sweetener, which is linked to the content of . In this study, for the first time in Croatia, pharmacobotanical, phytochemical and biological studies of stevia were conducted, in order to evaluate its potential biomedical application possibilities in prevention and treatment.

Aim: The aim of this study was to evaluate micromorphological and anatomical characteristics of the leaves and young stems, phytochemical composition, antioxidant effect, reducing power and total antioxidant capacity, ability to inhibit acetylcholinesterase and inhibitory effect on Src tyrosine kinase.

Materials and Methods: The plant material - Stevia rebaudiana (Bert.) Bertoni was cultivated in Croatia and collected in August 2012, just before flowering, and air-dried at room temperature. Identification of plant material was carried out at the Department of Pharmacognosy, Faculty of Pharmacy and Biochemistry. The presence of flavonoids, phenolic acids, tannins, , triterpenoids, sterols and essential oils were detected in methanolic leaf extract by thin layer chromatography. Contents of total polyphenols, tannins, flavonoids and phenolic acids in stevia leaves were determined spectrophotometrically. Antioxidant activities of ethanolic leaf extract were examined by different spectrophotometric methods in comparison with polyphenolic components and trolox as a reference. Effect of stevia and its individual bioactive components on Src tyrosine kinase was investigated by appropriate in vitro assay and compared with staurosporine.

Results and Conclusion: Our results showed strong antioxidant potential of stevia, mostly attributed to the phenolic compounds contained. Acetylcholinesterase (AChE) inhibitory activity of ethanolic leaf extract was evaluated in comparison with chlorogenic acid and galantamine as a reference. Stevia showed ability to inhibit AChE exceeding a 50% effect at 1000 μg/mL. It was also found that chlorogenic acid contributes significantly to its anti-AChE activity. This research showed that stevia leaves contain a significant amount of various bioactive components which demonstrated strong antioxidant activity and ability to inhibit acetylcholinesterase and Src tyrosine kinase. Obtained results suggested a great potential of stevia in prevention and treatment of various diseases and that is not just a plant that serves as means of sweetening food and pharmaceutical products. 6 EPSA Students' Science Publication Volume 3 | Edition 1 | October 2015

Q&A OF THE AUTHOR

General information about Ana Begić and Josip Barilar Both 25 years, Zagreb, Croatia, ana. [email protected], [email protected], University of Zagreb, Faculty of Pharmacy and Biochemistry

Why did you select this topic for your research? The stevia plant caught our attention when we realized that it has other beneficial biomedical properties besides being only a natural sweetener. As the population was using it more in the food industry, we wanted to prove that stevia also has a potential of prevention and treatment of various diseases.

How do you feel about having coped with the challenges that research brings? During a student research, confrontation with different challenges and problems are fairly common.

One should always try to approach difficulties rationally and try to use different methods and procedures to minimize the risk of getting bad results. Critical thinking in the process of research is crucial.

What skills did you develop as a researcher during your research period? During our research period we've learned to be more tolerant and more respective of people who dedicate their lives to science. We've also learned that a round bottom flask should not be placed on a flat surface, because you will lose the material you've been working on for weeks.

Would you like to share anything else with the students in Europe? Any recommendations? We'd like to advise them not to stick their heads in books and blindly believe what is written. In order to become a successful scientist it is of great importance to develop creativity because creativity is, along with knowledge, a core to scientific endeavour.

7 EPSA Students' Science Publication CELEBRATING ONE CENTURY AND THINKING ABOUT THE FUTURE: PRELIMINARY FINDINGS FOR A NEW MPHARM SYLLABUS AT LISBON FACULTY OF PHARMACY Madalena Plácido, Joana Catarino, Diogo Raimundo and Diana Ferreira Faculty of Pharmacy of the University of Lisbon, Lisbon, Portugal Supervisors: Afonso Cavaco, Maria Henriques and Andreia Bruno Others that contributed to the abstract: Diogo Simões and Francisca Lopes Introduction: The current curriculum of the Master degree in Pharmacy (MPharm) of the University of Lisbon is under an accreditation process during 2015. Induced by this evaluation, the Committee for the Centennial Celebrations of the Students’ Association (CENTAEFFUL) aimed to collect input from several parties involved in pharmacy education that could contribute to the improvement of the syllabus towards pharmacy labour opportunities. Methodology: The research project was divided into three phases. Phase one consisted of two focus groups, with six pharmaceutical professionals in each. The second phase was an online survey answered by a national sample of pharmacy students and pharmacists. The answers were collected through open and closed questions, including Likert scales to measure the agreement with sentences stated by professionals from the focus groups. The third phase was comprised of the presentation and discussion of the survey results at an Educational Forum (EF). The EF consisted of a two day debate using an amplification of survey results fed by live presentations of five experts in educational matters. The actual EF conclusions were accomplished through small group discussions between pharmacists, students and professors, complemented by a plenary session opened to the public. Results: The need to include internships within the first four years of the degree was mentioned in the focus groups. It was mentioned that having practitioners outside the university teaching some classes would be beneficial for students. Concerning the online survey, there were 473 answers. The majority of the respondents (51.8%) either ‘agreed’ or ‘strongly agreed’ that the current MPharm curriculum is not completely adapted to practice. Moreover, 77.8% strongly ‘agreed’ that students would benefit from internships within the first four years of the degree. EF conclusions supported the opinions previously mentioned and reinforced the need to introduce new mandatory subjects, such as non-prescription medicines pharmacotherapy, clinical trials and seminars about soft-skills. Introducing one more optional subject and increasing the use of real case studies during classes was also suggested. Conclusions: Most participants agreed with the need to adapt MPharm to the competencies required in the pharmaceutical labour market. Throughout the project several suggestions were able to be collected, which then were submitted to the Scientific Council, in order to support the changes that needed to be reflected for the proposal to be submitted for accreditation. Acknowledges: We would like to acknowledge the Pedagogical Council, the local and national Students’ Association (AEFFUL and APEF) and all professionals and students that contributed to this project. We are also grateful for the support of Merck Sharp & Dohme Portugal. EPSA Students' Science Publication Volume 3 | Edition 1 | October 2015

Q&A OF THE AUTHOR

General information about Madalena Plácido During 2014 a Committee for the Centennial Celebrations of the Students’ Association (CENTAEFFUL) was formed. This Committee aimed to analyse all Students’ Association journey: its past, present and future challenges. Focused on a more educational and scientific aspect, a Bureau for Education was born.

Why did you select this topic for your research? The current curriculum of the Master degree in Pharmacy (MPharm) of the University of Lisbon is under an accreditation process during 2015. As members of Bureau for Education we wanted to take this opportunity to analyze the adequacy of the MPharm syllabus to the labor market through pharmacy students and professionals’ eyes.

How do you feel about having coped with the challenges that research brings? The main challenge was to define the right modus operandi for our research. Luckily we benefited from the help of professionals and faculty professors, to whom we are deeply grateful. Another challenge was the gap between academy and practice, which we tried to shorten through this investigation work.

What skills did you develop as a researcher during your research period? This kind of research allowed us to develop our critical thinking and data analysis abilities. We also improved our communication and organizational skills. Moreover, our awareness about the current MPharm syllabus and the labor market needs also increased.

Would you like to share anything else with the students in Europe? Any recommendations? Research is not only discovering new drugs or a cure for a disease. If you believe that you can make a difference in your own university, go ahead! Understand the opinion of professionals from different areas, professors and your colleagues. Certainly you will learn something new and contribute to improve your university.

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THE INFLUENCE OF ACUTE PAIN ON OSMOTIC PRESSURE AND CORTISOL CONCENTRATION IN TEARS Maja Lukić and Matea Ćurčić, Faculty of Pharmacy and Medical Biochemistry, University of Zagreb, Croatia

Introduction: Tears are far more than just a fluid on the surface of our eyes. Consisting of three distinct layers, they are essential for eye protection, lubrication and nourishment as well as maintaining health of the eyes and optical function of their surface and for providing clear vision. Furthermore, tears prevent eye infections by continually removing foreign matter thus, keeping the surface of the eyes smooth and clear. Tear production is controlled both by the central and autonomic nervous system. While basal tears’ production is under control of the accessory lacrimal glands, reflex lacrimation is mediated by the neural reflex arc that stimulates the secretion of the main lacrimal glands. Pain causes stress to the organism and therefore, production of stress mediators is stimulated. Cortisol is, along with DHEA, adrenalin and noradrenalin, one of the four main stress mediators. Although the origin of cortisol in tears is not known yet, it is assumed it might derive from the lacrimal gland. The final result of the dynamic changes of the lacrimal fluid is determined by measuring the osmolarity of the lacrimal fluid.

Aim: In this study, the applicability of tears in the diagnostics of acute stress was evaluated. The main objectives of the study were to evaluate cortisol values and osmotic concentration in tears before and after stimulus and whether a consistent answer to stress could be diagnosed in tears.

Materials and Methods: The study included 18 volunteers (10 women and 8 men) whose tears were collected and analysed at the Clinical Hospital Center 'Sestre Milosrdnice'. The tear samples were collected before and after nasal hair trimming. The concentration of cortisol was measured using ELISA whilst osmometry was used as a method to determine the osmolar concentration.

Results and Conclusion: Basal cortisol values in men showed good correlation with osmolarity in tears. After acute stress, this correlation was lost, which can be due to the higher sensitivity to pain in males. After stimulus, cortisol and osmolar concentration decreased due to the dilution caused by tear production. Osmolarity and cortisol showed a moderate correlation before the painful stimulus, which is completely lost after the stimulus. Pain stimulus at the root hairs activates the sympathetic nervous system and its possible effect on the secretion of glands.

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Q&A OF THE AUTHOR

General information about Maja Lukić 27 years old, Osijek, Republic of Croatia E-mail: [email protected] The faculty of Pharmacy and Medical Biochemistry (Graduated 2015), University of Zagreb

Why did you select this topic for your research? The idea of the topic came spontaneously but from the very beginning Matea and I knew we wanted to conduct an investigation on tears, as they as a sample are rarely used in clinical practice.

How do you feel about having coped with the challenges that research brings? From the very beginning, there were a lot of challenging aspects of our investigation. Firstly, tears collection and limited sample volume, as well as analyte measurement in a low concentration range. Moreover, there was no available literature on studies of human tears. Despite all, we did not give up and succedeed in obtaining our results.

What skills did you develop as a researcher during your research period? Above all, patience, organizational skills, accuracy and punctuality.

Would you like to share anything else with the students in Europe? Any recommendations? I strongly recommend conducting an investigation during their studies. In this way, you can gain great experience in the field of your study and through cooperation with your colleagues, gain new friends.

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EUROPEAN FEDERATION FOR PHARMACEUTICAL SCIENCES (EUFEPS)

The European Federation for Pharmaceutical Sciences is a voluntary federation of national associations and societies of pharmaceutical scientists, established in 1991 to advance research in the pharmaceutical sciences in Europe. This can be achieved by promoting cooperation between national, regional and European societies or associations which aim at the advancement of pharmaceutical sciences, and by promoting cooperation between and with other pharmaceutical organisations and between individual pharmaceutical scientists.

The Mission EUFEPS exists to help to meet the challenges and seize the opportunities created by the consolidations occurring both within Europe and globally, driven on by a combination of rapid advances in science and technology, economic pressures, and by political will. Within this frame, EUFEPS’s mission is to advance sciences for better medicines and health and serving the pharmaceutical sciences and innovative drug research in Europe. Spearheading a number of initiatives, EUFEPS works with its membership, throughout many nations in Europe.

European Dimension EUFEPS is unique being the only pan-European organisation that represents, under one umbrella, all the pharmaceutical sciences and pharmaceutical scientists engaged in drug research and development, drug regulation and drug policy making. The existence of such an umbrella platform facilitates the highly innovative, integrative and interdisciplinary approaches that are essential if we, in Europe, are to deliver to our citizens safe, effective, economic and timely medicines. The ultimate benefits are an improving health, quality of life, and wealth of our continent.

EUFEPS is recognised by the European Commission, as representing the integrative pharmaceutical sciences within Europe. EUEFPS is also recognised by the EMEA as a neutral scientific resource for independent opinions on draft regulatory guidelines, while EUFEPS works with other European organisations, such as EFPIA, to help identify and promote training to meet industrial needs. EUFEPS has the ambition to provide a forum for policy development in the pharmaceutical sciences, particularly in relations to the discovery and development of new drugs and their introduction into the market, but also as to medicines usage. This includes policies on and leadership development as to: research, education & training, profiling and regulatory affairs.

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In addition, EUFEPS plays an active and influential role also in the global arena. It is recognised by the USA FDA and it works actively with its sister organisation AAPS to develop co-sponsored meetings and workshops that run alternatively in Europe and the USA, and is developing links with Asian scientists. Through involvement with its Board of Pharmaceutical Sciences, EUFEPS is also working with FIP to advance the pharmaceutical sciences globally.

UPCOMING CONFERENCES ORGANISED OR CO- ORGANISED BY EUFEPS

8th International Symposium on Microdialysis Preclinical, Clinical, Analytical 25-27 May 2016, Uppsala, Sweden

EUFEPS Annual Meeting 2016 Clinical Outcome and Regulation of Advanced Drug Delivery Products 13-15 June 2016, Istanbul, Turkey

8th Pan-European Science Conference on QbD and PAT Sciences Inventing Tomorrow’s Development and Manufacturing 3-4 October 2016, Cork, Ireland

2nd Conference on the Global Bioequivalence Harmonisation Initiative Autumn 2016, Washington DC, United States of America

For more information see conferences at: www.eufeps.org

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WHAT DOES EPSA OFFER IN THE FIELD OF SCIENCE?

Next to the EPSA Students' Science Publication (ESSP) that you are reading now, EPSA also offers two other projects in the field of Science: the Science Day and the Science Excursion.

Science Day The Science Day is an annual educational event that takes place during the EPSA Annual Congress and is dedicated to students’ scientific work. Participants are given the opportunity to present the scientific research they have conducted orally and through a poster exhibition. This year fourteen students in total participated in one or two parts of the event. A jury, consisting of the European Federation for Pharma- ceutical Sciences, EUFEPS representative, a Pierre Fabre Lab. Representative and the EPSA Science Coordinator Successor, assessed the presentations. Since the competition this year was extremely tough, the jury agreed on two winners for the oral presentation.

Science Excursion The idea of the Science Excursion is to bring students closer to science and to broaden the Congress experience to an interactive and local surrounding. It is a project that is planned to be held at all major EPSA events.

The upcoming excursion will be held during the Autumn Assembly in the restored oldest pharmacy in the Santo Spirito Hospital in Malta. Participants will visit the pharmacy museum and witness a compounding demonstration.

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