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1st Edition 2021 - 2022
POLICY NEWSLETTER
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Table of contents
Dear readers,
4 Cancer medicine prices in 30 european countries
Before you lies the 1st edition of EPSA Students Science Publication (ESSP) Volume 9. ESSP is an EPSA Scientific Project which provides students with the opportunity to share their research findings, in abstract format, on an international level. The abstracts are reviewed by professionals from the European Federation for Pharmaceutical Sciences (EUFEPS) and provided with insightful feedback. Students are encouraged to improve their academic writing skills.
6 Antihyperglycemic potential of hemp extracts (Cannabis sativa, Cannabaceae) 8 Targeting autophagy complements the action of ibrutinib and venetoclax in chronic lymphocytic leukemia cells 10 New approach in determining gentamicin in plga-composites by high performance liquid chromatography (hplc) and photo diode array (pda) detection
This edition contains four abstracts from different fields of pharmaceutical sciences. You will gain insight into price differences of cancer medicines over the world and read about antihyperglycemic potential of cannabis. Furthermore, you can discover the action of ibrutinib and venetoclax in targeting autophagy complements. And a new approach in determining gentamicin in PLGA composites by high performance liquid chromatography and photodiode array detection will be discussed. The abstracts represent the effort and time the students have put in to conduct amazing research projects all over Europe. It was a pleasure to collaborate with the students and read their abstracts. Many thanks to students who gathered their courage and sent their research abstracts, and for sharing your work so that our pharmaceutical knowledge will broaden. I would like to express my gratitude to the European Federation for Pharmaceutical Sciences for the effort and time they have put into reviewing the abstracts, supporting in the making of this publication, and giving insightful feedback to the students. Without the publication department, this edition would not have been possible. Thank you for your hard and dedicated work. I hope you enjoy reading this. Yours in science, Yong Xin Cao EPSA Science Coordinator 2021/2022
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European Pharmaceutical Students’ Association
Cancer Medicine Prices in 30 European Countries Author: Neda Zare, Naheem Shazad, Umar Ali, Sheikh Malik, Besime Ozbek, Nancy Dike, Zaheer-Ud-Din Babar Scientific Coordinator: Professor Zaheer-Ud-Din-Babar Institution: Department of Pharmacy , University of Huddersfield, HD1 3DH, United Kingdom
INTRODUCTION: Cancer is one of the leading causes of death worldwide, and it accounted for almost 10 million deaths around the globe in 2020. The number of deaths attributed to cancer is increasing across the world. High pharmaceutical product prices constitute a significant challenge both in high and low-income countries.
Log ex-factory prices per unit indexed
AIM: We aimed to survey the prices of cancer associated with ex-factory unit price at the 5% significance level for Ibrance (p=0.001 medicines in 30 European countries. for location; p<0.001 for GDP per capita). MATERIALS AND METHODS: Based on our Medicine strength was significantly associated previous experience we exUsing the Wilhelmy with the ex-factory unit price at the 5% plate tensiometric method, the interfacial significance level for several drugs: Afinitor, tension between oil and water was measured Caprelsa, Keytruda, and Zytiga (p<0.001 in all with the addition of different concentrations cases). of selected emulsifiers. Emulsifiers were prepared based on the required HLB value, CONCLUSION: Our results suggest variation in namely Tween 40 HLB=15.6; mixtures of ex-factory price per unit of cancer medicines Tween 40 and Span 85 with HLB=7 and in 30 surveyed countries. The price data 12, and Solutol HS15 HLB=14-16. With were not available for all drugs in all surveyed the selected concentrations of emulsifiers, countries. Our findings may help policymakers emulsions with different proportions of oil to develop appropriate policy interventions to and water were prepared (1:3 oil:water, 1:3 promote price transparency and equity. water:oil, and 1:1). Emulsions were prepared using a rotor-stator homogenizer under the same conditions of 15000 rpm for 10min. Droplet size was monitored by an optical microscope, and viscosity by a cylindrical viscometer. The stability itself was determined organoleptically. 80
15 0
m g
ha Zy rd tig c a Zy 25 aps Af tig ul 0 in e a ito mg 50 ta r2 0 Im bl m . 5 et b gf Ca ruv ilm mg ica ta pr c b el o 14 le at sa t ed 10 0 m ta gh 0 bl m ar e gf ilm d ca t ps co ul at Af e e in Ca ito d ta pr r5 bl el Af et sa in ito mg Ta 300 t r1 ab gr le 0 iss mg t m f o g 40 ilm Ta co tab gr le iss mg at t ed f o 80 ilm co tab m l a et gf Ib ilm ted ra nc co tab e le at Ib 75 t ed ra m nc ta gh e bl 10 ar Ib et d 0 ra c m nc g h aps e ul 12 ar e d 5 ca Ke mg ha psu yt ru le rd d Ke a 50 cap su yt m le ru da g po w 25 de m r g/ m l4 m l
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Al ec en sa
RESULTS: Austria, Germany, Sweden, and Spain ranked high priced countries, while Bulgaria and the UK were at the lower end. The difference between the highest-priced country (Germany) and the lowest-priced countries (Czech Republic & Sweden) varies between 24.98% and 170% for the same medicine. Inferential statistics revealed that location and GDP per capita were significantly
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Drugs
Figure 1. Boxplot of drug prices (ex-factory price per unit) indexed (price in the lowest priced country=€10), as of May 2017 to April 2019, in 30 European countries. The box displays the interquartile range (IQR); the bottom and top of the box are the 25th and 75th percentiles (the 1st and 3rd quartiles, respectively), and the band near the middle of the box is the median. The lines describe the bottom and top whiskers.
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Questions & answers Please, tell us a little bit more about yourself. I am a fourth-year pharmacy student at the University of Huddersfield. My passion is healthcare and anything that can relieve people from worrying about accessing treatment when they’re ill. I migrated to England in 2015, and after two years of college, I succeeded to earn a place at Huddersfield University. It was my dream to be a pharmacist, and since I started my course, the more I know, the more I fell in love with it. I studied nursing in Iran, and I witnessed how people can struggle to access medicines that are already there, but the price is the main burden to them. This applies to many countries worldwide. Tell us a bit more about your research and its significance. My dissertation was on cancer medicine prices in 30 European countries. You might be surprised to know the same cancer medicine with the same brand from the same company has a price difference between countries up to 170%. In conclusion, this massive variation and the high price of cancer medicines are a burden to access treatment in some countries. As the number of cancer incidences increases, this issue can get more severe and affect more people. We tried to show this based on evidence and data available to inform policymakers of the significance of their decision about pricing cancer medicines. What was the biggest challenge while carrying out the research and how did you overcome that? Our data were from 30 European countries in their original currency from different periods
from May 2017 to April 2019 inclusive; equalizing these currencies was a challenge. Also, for some medicines, data were not available in all 30 countries, such as Malta. This was resolved by consulting various studies and methodological approaches, and the best way to go about it. In your opinion, what is the benefit of joining ESSP and what advice do you have for students undertaking research in the future? Each research that you conduct is a voice that you want to be heard. ESSP is an excellent platform for students to make this happen and this is an excellent opportunity to represent their university. For me getting my abstract published with ESSP is like hard work paid back, and it’s a fantastic experience to have. For those who are researching the future, I would say first: - Conduct an extensive literature review. - Listen to the expert advice and trust your supervisor’s guidance. - Make your voice to be heard and your hard work to be seen and use all the supports and opportunities you have
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European Pharmaceutical Students’ Association
ANTIHYPERGLYCEMIC POTENTIAL OF HEMP EXTRACTS (Cannabis sativa, Cannabaceae) Author: Marija Perić Scientific Coordinator: Nebojša Kladar, PhD, Assistant Professor; Nebojša Salaj, MPharm, Junior Researcher Institution: University of Novi Sad, Faculty of Medicine, Department of Pharmacy
INTRODUCTION: Hemp (Cannabis sativa, Cannabaceae) is a plant species which is primarily known for its high content of cannabinoids, while phenolics and flavonoids present in hemp have been less studied. Diabetes mellitus type 2 (DM2) is a metabolic disorder characterized by chronic hyperglycemia and shows significant increase of incidence in the world. Phenolics and flavonoids possess antioxidant effect, but there is also possibility to exert antihyperglycemic effect by reduction of dietary carbohydrate digestion, and consequent absorption, through the inhibition of key enzymes involved in their intestinal metabolism – α-amylase and α-glucosidase. AIM: The aim of the conducted study was to examine antihyperglycemic potential (inhibition of α-amylase and α-glucosidase) of hemp aqueous and ethanolic extracts in vitro and perform their preliminary and detailed chemical characterization. MATERIAL AND METHODS: Aqueous and ethanol extracts were preliminary and detailed chemically characterized by spectrophotometric and high performance liquid chromatography methods. The potential of the extracts to inhibit the activity of α-amylase and α-glucosidase was also estimated. RESULTS: Preliminary chemical characterization showed high content of phenolic and flavonoid compounds in the obtained extracts. The results of detailed chemical characterization showed the absence of gallic acid, quercetin and rutin and the presence of caffeic acid, chlorogenic acid and trans-cinnamic acid. The aqueous extracts showed higher potential to inhibit α-amylase compared to the ethanol extracts, which were more potent inhibitors of α-glucosidase.
CONCLUSION: Both types of hemp extracts contain high amounts of phenolic and flavonoid compounds, but there are some content variations of these extracts. The aqueous extracts are more potent α-amylase inhibitors, while the ethanolic extracts show higher potential to inhibit α-glucosidase in the used in vitro test-systems, when compared to the ability of acarbose to inhibit aforementioned enzyme. However, future in vivo research is required for further understanding of the antihyperglycemic activity mechanisms of secondary metabolites found in examined hemp extracts.
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Questions & answers Please, tell us a little bit more about yourself. My name is Marija and I am a 5th year Pharmacy student. There are so many reasons why I have chosen pharmacy, but the most important is being able to help people of all ages every day and also be there to support them about their therapy. I am interested in biochemistry, toxicology and clinical pharmacy and I really enjoy working in the lab, spending time with my family and friends and listening to music. Tell us a bit more about your research and its significance. For the past few years hemp has been in the spotlight because of its anticancer effect, but we wanted to discover other effects. The reason why we have chosen Diabetes Mellitus type 2 is the increase of its incidence in the world. The significance of our research is that we have found out that hemp extracts are showing some antihyperglycemic potential so we are hoping we have motivated colleagues to continue research on this topic. What was the biggest challenge while carrying out the research and how did you overcome that? The biggest challenge while carrying out the research was staying in lab every other day for so many hours despite of all my students responsibilities and private ones, but I have founded a purpose and it was all worth it. Everything about this research was beautiful and meaningful experience.
In your opinion, what is the benefit of joining ESSP and what advice do you have for students undertaking research in the future? In my opinion, the benefit of joining ESSP is having an opportunity to present your research and be able to get in touch with colleagues. My advice for students is just be curious and never give up because you will never know what is waiting for you in the end. Maybe you will change the future.
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European Pharmaceutical Students’ Association
TARGETING AUTOPHAGY COMPLEMENTS THE ACTION OF IBRUTINIB AND VENETOCLAX IN CHRONIC LYMPHOCYTIC LEUKEMIA CELLS Authors: Alma Tana Jakoš Djordjevič Scientific coordinator: prof. dr. Irena Mlinarič-Raščan, Damjan Avsec Institution: University of Ljubljana, Faculty of Pharmacy, Chair of Clinical Biochemistry
INTRODUCTION: Chronic lymphocytic leukemia (CLL) is the most common leukemia in the Western world. In the recent years, BTK inhibitor ibrutinib, PI3K inhibitor idelalisib, and Bcl-2 antagonist venetoclax revolutionized the treatment of CLL. However, these therapies present with several drawbacks, such as severe adverse events and emergence of resistance. Therefore, the demand for novel targets is high. One of such targets is autophagy, a protective and degradative cell process that plays an important but still fairly undefined role in CLL. AIM: To elucidate the antileukemic potential of mechanism of cell death. The mean EC50 values of MRT68921 in patient-derived CLL targeting autophagy in CLL. cells (n = 13) were 3.6 μM and 2.9 μM after MATERIAL AND METHODS: Autophagy 24 and 48 hour treatment, respectively, reporter cell line THP-1 Difluo hLC3 and demonstrating that MRT68921 acts in a imaging flow cytometry were used to monitor concentration and time-dependent manner (p autophagic flux. Autophagy was blocked using = 0.0036). Moreover, compared to peripheral ULK1/2 inhibitor MRT68921. Cytotoxic and blood mononuclear cells MRT68921 was synergistic effects of MRT68921 and targeted more cytotoxic to CLL cells (p = 0.0013), therapy in primary CLL cells (the study was indicating it acts selectively cytotoxic towards approved by Slovenian National Medical malignant B cells. Ethics Committee; 0120-136/2019/4) were evaluated using PrestoBlue assay. To identify CONCLUSION: In the present study we the mechanisms of synergistic effects, PI demonstrated that autophagy inhibition uptake and cell cycle assay were performed with ULK1/2 inhibitor MRT68921 acts selectively cytotoxic towards malignant B using flow cytometry. cells and augments the action of ibrutinib RESULTS: Using autophagy reporter cells, and venetoclax. Thus, we recognized we showed that ibrutinib and venetoclax but autophagy as an important target and not idelalisib induce autophagy. Blocking the inhibition of autophagy as a promising induction of autophagy with ULK1/2 inhibitor new approach for the treatment of CLL MRT68921 augmented the cytotoxicity of ibrutinib and venetoclax in CLL cell line MEC-1 and primary CLL cells. Furthermore, MRT68921 alone was able to cause cell cycle arrest at G2/M phase, but in combination with ibrutinib and venetoclax, it acted synergistically cytotoxic through a caspase-dependent mechanism, indicating that apoptosis is the
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Questions & answers Please, tell us a little bit more about yourself. My name is Alma and I finished my Master’s degree in Pharmacy at University of Ljubljana in April 2021. I am currently living in Brussels, where I am doing a one-year Masters in International Business Economics and Management. In the near future, I plan to use my knowledge in business to realise some of my scientific research projects that I care about most. Tell us a bit more about your research and its significance. In our research, we recognized autophagy as a new therapeutic target in chronic lymphatic leukaemia (CLL) and inhibitors of autophagy as a potential new drug for the treatment of CLL. The results of our research thus represent an encouraging first step for further research into autophagy inhibitors in the treatment of CLL. Our findings are relevant to patients for whom existing therapy is inappropriate or ineffective. The prevalence of CLL is rising due to our aging population, making this research ever more important. What was the biggest challenge while carrying out the research and how did you overcome that? The biggest challenge was the all-consuming nature of the experiments. After I treated the cells with different substances, I had to wait for precise intervals of between 24 and 72 hours to take measurements. The smallest mistake or contamination meant I had to do the experiments all over again and prolong results by a matter of days.
In your opinion, what is the benefit of joining ESSP and what advice do you have for students undertaking research in the future? I think that ESSP is a great starting point for young researchers who are starting their professional career. After investing a great deal of time and energy in your research, it is incredibly rewarding to be able to share it with a wider audience and achieve international recognition. My advice to future students is to choose a research project they are really passionate about and to take advantage of opportunities to share their work and be heard.
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European Pharmaceutical Students’ Association
NEW APPROACH IN DETERMINING GENTAMICIN IN PLGA-COMPOSITES BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) AND PHOTO DIODE ARRAY (PDA) DETECTION Author: Simona Maria Bulea, Dr. Costel-Valentin-Manda, Dr. Maria-Viorica Ciocîlteu Institution: University of Medicine and Pharmacy in Craiova, Faculty of Pharmacy, Craiova, Romania
INTRODUCTION: Gentamicin is an aminoglycoside antibiotic (a mixture of four major compounds) used in the treatment of bacteria resistant to other antimicrobial agents involved in bone infections, but it has severe adverse effects (ototoxicity, nephrotoxicity). The inclusion into a polymer, polylactic-co-glycolic acid (PLGA) microspheres and the local use of the drug largely reduce these side effects. Determination of gentamicin in the previously synthesized PLGA-gentamicin composites is important to assess loading efficiency of the antibiotic. . AIM: The objective was to elaborate and 0.75 mL borate buffer and 0.50 mL FMOCvalidate a simple HPLC method appropriate in Cl solution (for 5 minutes). 200 μL were next terms of time analysis, robustness, precision treated identical with the calibration curve case. and accuracy. MATERIAL AND METHODS: The instrument used was a Thermo Finnigan HPLC System with a PDA detector. Separation was achieved using a C18 chromatographic column (dimensions 250 x 4.6 mm internal diameter and 5 μm particle size). Elution was performed in isocratic mode with acetonitrile: methanol: water (85:10:5, v/v) at 1 mL/min flow rate. The discrete wavelength used in the quantitation process was 265 nm. To perform the calibration curve, gentamicin standard solutions (0.05 - 1 mg/mL) were prepared in a borate buffer (pH 9.7). A derivatization reaction is needed to increase gentamicin detection. 1 mL standard solution was added 0.50 mL solution of 9-fluorenylmethylchloroformate (FMOC-Cl) as a derivatizing agent. 200 μL were added 800 μL mobile phase and 20 μL were injected into the HPLC. Gentamicin analysis in PLGA microspheres was performed on 14 mg composite. 1 mL borate buffer was added (kept for 12 h to achieve hydrolysis of PLGA). 0.25 mL of the resulting solution was filtered through a 0.45 μm porous membrane, then mixed with
RESULTS: Within this method, the regression equation for target concentration range was linear (correlation coefficient “R” was 0.998 and 0.996 for peaks 1 and 4, respectively). Run-time was short (15 minutes) in contrast to other HPLC methods. This is the reason peaks 2 and 3 are not fully separated. Precision and recovery were within ±5% limit. Gentamicin percentages in different microspheres were found between 0.64-2,85%. CONCLUSION: The presented method is suitable for gentamicin analysis in PLGAcomposites. Both validation data and sample analysis qualify this method to determine loading efficiency of gentamicin in the microspheres.
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Questions & answers
What was the biggest challenge while carrying out the research and how did you overcome that? The most important challenge was properly performing the derivatization reaction. Usually, this kind of reactions are incomplete and highly non-reproducible. The choice of pH is very important. Adjusting the pH value to 9.7 (with borate buffer) was necessary to ensure a better reaction between the amino- groups of gentamicin and FMOC-Cl. Investigation of reaction time was also important. Optimal time for the derivatization reaction was found to be 5 minutes. Longer reaction times did not provide any increase in chromatographic peak areas for the target compounds of gentamicin.
Please, tell us a little bit more about yourself. I am a final year student at the Faculty of Pharmacy in Romania. Also, I have been a volunteer in my local pharmacy student association since my first year of university .Since highschool I was passionate about chemistry and I knew that I wanted to bring a contribution to the health industry through research.In my leisure time I enjoy travelling, In your opinion, what is the benefit of joining painting and cycling. ESSP and what advice do you have for students Tell us a bit more about your research and its undertaking research in the future? Joining ESSP is a great opportunity for significance. This HPLC method was part of a more students to publish their research,start their complex study using various methods, which journey as scientists and to improve their involved synthesis and physico-chemical resume.My advice for students undertaking characterization of the resulted polylactic-co- research is to choose a subject they like,have glycolic acid (PLGA) composites. The double patience and perseverance and to seek emulsion synthesis method was chosen, advice from their mentor whenever they need being the most suitable for encapsulating it. water-soluble drugs, such as gentamicin. For qualitative analysis of the composites, Fourier Transformed Infrared (FT-IR) spectroscopy, Dynamic Light Scattering (DLS) and X-Ray Diffraction (XRD) methods were employed. For the quantitative assay of gentamicin, a new HPLC chromatographic method was developed, consisting in an initial treatment of gentamicin with a derivatizing agent (FMOCCl), followed by photo-diode array (PDA) detection of the derivative. Derivatization is required because of the lack of chromophores in gentamicin compounds. This method proved to be suitable for gentamicin quantitative analysis in the synthesized PLGA microspheres, according to validation data presented.
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European Pharmaceutical Students’ Association
The Science of Cosmetics October Edition 2021
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