Browsing by master's degree program "Master's Programme in Pharmacy"

The objective of this research has been to investigate the management of alerts of Medicines Verification Systems in Europe. Verification of medicines according to Falsified Medicines Directive (FMD) came into force 9.2.2019. There is no standardized tool or system for the management of alerts, every Medicines Verification Organisation and manufacturer have had to find their own ways to manage the alerts. The research has been performed as a theme survey via questionnaire that has been sent to Medicines Verification Organisations in 30 European countries. Information to the questions of the questionnaire has also been gathered from the Internet pages that are mainly maintained by the Medicines Verification Organisations. This kind of method triangulation has been used in order to improve the reliability of the research. Answering rate of the survey was 17 %. By including the information gathered by method triangulation the overall yield percent of information in this study was 45 %. The information received via the questionnaire did not contradict with the public information. As conclusion, marketing authorization holders have been registered as users of the National Medicines Verification Systems or they have signed a contract with Medicines Verification Organisations. Marketing authorization holders are paying the costs of the Medicines Verification Systems. Penalties of FMD non-compliance are in use in part of the European countries. In the beginning of the implementation of the Medicines Verification System there has been stabilization periods in use which have already ended in half of the European countries. National competent authority is informed about system alerts typically in case of suspected falsification. In half of the European countries there is a separate computerized alert management system in use. Marketing authorization holder usually has access to the system. In some of the countries it is possible to integrate the system to the own serialization system of the marketing authorization holder. In six European countries there has been set a specific time for the alert investigation of the marketing authorization holder. Based on the results of this research the alert management system that covers the whole Europe that European Medicines Verification Organization is planning would really be needed. One common computerized system and common rules would ease up the alert management of all the stakeholders of the medicines verification.

The Finnish medicine reimbursement system is complex and several different conditions required by the Health Insurance Act (1224/2004) and the Social Insurance Institution of Finland (Kela) must be met in order to receive medicine reimbursement. The understanding of medicine reimbursement criteria from the perspective of medicine users has not been studied in Finland before, and little research has been done on the subject internationally. Medicine user-oriented research on the medicine reimbursement system, both in Finland and internationally, has largely focused on the financial opportunities of medicine users to purchase medicines and their opinions on the fairness of medicine reimbursements. The aim of this study was to obtain information on the understanding of medicine reimbursement criteria and the background factors affecting it, the implementation of price, generic substitution and medicine reimbursement counselling in pharmacies, seeking advice on medicine reimbursement, and the financial difficulties of buying prescription medicines. The material used in this study was from the population survey (n=1650), which examined the activation of price competition for pharmaceutical products and customers' expectations of pharmacy operations. The understanding of medicine reimbursement criteria, the price counselling provided in a pharmacy, the effect of financial challenges on the non-buying of medicines and the use of sources of advice related to the medicine reimbursement were described as frequency distributions. The effect of background factors on the understanding of medicine reimbursement criteria was compared using the chi-square test and logistic regression analysis. About a third (31%) of respondents told that they do not understand the basis on which medicine reimbursement is usually received for prescription medicines, and 13% had unclear why they had not been reimbursed for their prescription medicine in the past year. Especially younger age, low income, low medication use, depression and other mental health problems, lack of long-term illness, and insufficient medicine reimbursement counselling in a pharmacy were found to be associated with poorer understanding of medicine reimbursement. 72% of the participants in the study felt that they usually receive sufficient information about the prices of medicines and 61% about the reimbursement of medicines when buying prescription medicines from a pharmacy. Less than half (47%) of respondents felt that they have usually received sufficient information about how the amount of reimbursement for medicines is determined. Slightly over 70% of respondents said that they are usually told about the cheapest medicine available when buying a prescription medicine and/or are suggested to switch to a cheaper one. About 60% were usually told about the difference between the two interchangeable medicines. 88% of respondents would seek information about medicine reimbursement primarily from a pharmacy or pharmacy´s online services. About 3% of all respondents in the study had not bought a medicine prescribed by a doctor for financial reasons in the last six months. Based on the study, about a third of medicine users have remained unclear regarding medicine reimbursements, and not everyone feels that they have received sufficient counselling and information about medicine prices and medicine reimbursements when buying prescription medicines from a pharmacy. Counselling from a pharmacy was found to be related to understanding of medicine reimbursement criteria. Advise on the pricing, medicine reimbursement and generic substitution should continue to be actively provided to medicine users, so that the counselling meets the requirements of the law and the knowledge of the reimbursement system of medicine users can be improved. Based on the results of this study, counselling should be targeted in particular at younger, low-income and from mental health problems suffering medicine users, as well as those who are less familiar with reimbursement issues, for example due to low morbidity or medicine use.

Good availability of medicines means that authorised medicines are placed on the market and not in a short supply. Drug shortages have increasingly become a common problem that has compromised the continuous availability of medicines. Drug shortages are caused by many complex factors, such as capacity constraints, manufacturing difficulties, business decisions, availability of raw materials, and sudden increase in demand. Drug shortages can cause adverse effects, medication errors, allergies, and delays in necessary treatments. There have been studies that have explained the reasons behind medicines shortages. However, more information is needed especially from the perspective of markets in Finland. The aim of this study was to reinforce and deepen the knowledge concerning the availability of medicines in Finland. The aim was to identify the group of medicines that are more exposed in short supply than others and define the root causes of medicine shortages. In addition, the aim was to form an overview of the availability of medicines in Finland. The study was based on a Finnish medicine agency's registry. Data were collected retrospectively from materials that contained shortage notifications from marketing authorisation holders and mandatory reserve supplies permissions for exemption to maintain lower stock levels. The study was dated between the years 2017 and 2020. More detailed data from the medicine shortage was only available from the year 2020 because of changes in procedures. The study also contained data from the register for all marketed and non-marketed medicinal products with marketing authorisation. The data classified with ATC-codes, because it is used internationally and thus makes the study to comparative to other studies. Data were analysed with cross-tabulation and frequency distributions. The study addressed that drug shortages were reported of medicines that are commonly used in Finland, such as nervous and cardiovascular system drugs. These medicines covered almost half of the annual shortage notifications. The number of drug shortage notifications has increased annually by approximately 40 percent. Correspondingly mandatory reserve supplies permission for an exemption to maintain lower stock levels were reported most on nervous system drugs and anti-infectives for systemic use. The number of annual permissions remained quite constant. The amount of the permissions increased 14 percent between the years 2017 and 2018 and a further 6 percent to the year 2019. However, the amount of permissions increased 26 percent in the year 2020. There is clearly recognized the effect of the COVID-19 pandemic when considered the rate of shortage notifications and mandatory reserve supplies permissions. The main reasons for the shortages were capacity constraints (32%) and increased demand (21%). Most of the drugs in short supply were drugs with national (33%) or decentralised (30%) marketing authorisation procedures. Broadly were able to state that the availability of medicines was at an acceptable level. 62 percent of all medicinal products with marketing authorisation were placed on the market. In addition, only 29 percent of older drugs with marketing authorisation accept during the years 1996 to 2003 were withdrawn from the market. In future, more large-scale studies are needed based on this study to improve the system that maintains the continuous and high-quality medicine distribution. Most important is to create a better tracking system and co-operation between national and international officials.

DNA-origamit ovat pitkästä DNA-juosteesta laskostettuja nanorakenteita, jotka voidaan suunnitella monen muotoisiksi ja kokoisiksi. DNA-origamit ovat muodostuneet luontaisesti biohajoavasta materiaalista, joten ne sopivat hyvin biolääketieteellisiin tarkoituksiin, kuten lääkekantajiksi. Aiemmissa tutkimuksissa on kuitenkin huomattu, että ihmiskehon entsyymit voivat hajottaa DNA-origamin rakenteen. DNA-origamien lipidipäällystäminen on yksi keino suojata rakennetta tällaiselta entsymaattiselta hajoamiselta. Tässä tutkimuksessa DNA-origameja päällystettiin kationisilla DOTAP (1,2-dioleyyli-3-trimetyyliammonium-propaani) lipideillä. Tutkimuksen tavoitteena oli tutkia ja optimoida päällystämisprosessia. Tutkimuksessa testattiin erilaisten inkubaatioaikojen ja -lämpötilojen sekä ionisten olosuhteiden vaikutusta muodostuvaan lipidipäällystykseen. Lipidipäällystettyjä DNA-origameja altisteettiin sonikaatiolle ja ekstruusiolle, jotta havaittaisiin, voisivatko nämä menetelmät häiritä muodostunutta lipidipäällystettä. Neutraalia DOPC (1,2-dioleoyyli-sn-glysero-3-fosfokoliini) lipidiä käytettiin tutkimaan, kuinka erilaiset lipidikombinaatiot vaikututtavat päällystyksen muodostumiseen. Työssä käytettyjä tutkimusmenetelmiä olivat dynaaminen valonsirontamenetelmä, agaroosigeelielektroforeesi ja transmissio-elektronimikroskopia. Kokeissa lipidipäällystäminen onnistui ionittomissa olosuhteissa ja 30 minuutin inkubaatioaika huoneenlämmössä oli riittävä päällystyksen muodostumiselle. DNA-origami-partikkelit olivat täysin päällystettyjä, kun käytettiin stoikiometristä suhdetta nDOTAP/nORIGAMI ≈ 20 000. Tällöin lipidipäällystetyt DNA-origamit muodostivat mikrometrien kokoisia rykelmiä. Kun käytettiin stoikiometristä suhdetta nDOTAP/nORIGAMI ≈ 10 000, TEM kuvissa oli havaittavissa päällystämättömiä ja satojen nanometrien kokoisia päällystettyjä DNA-origameja. Kylpysonikaation havaittiin häiritsevän aggregoituneita lipidi-DNA-partikkeleita muodostaen pienempiä partikkelirykelmiä. Kun käytettiin DOPC lipidejä, huomattiin, että ne eivät muodostaneet yhtenevää lipidipäällystettä DNA-origamien ympärille. Kokeissa ionisten olosuhteiden havaittiin epästabiloivat DNA-origamien päällystysprosessia, joka johti partikkelien kokojakaantuman lisääntymiseen. Tässä työssä DOTAP lipidejä käytettiin päällystämään DNA-origameja ja lipidimäärää kasvattamalla saatiin muodostettua monesta lipidikerroksesta muodostuneita lipidi-DNA-komplekseja. Sonikaation havaittiin olevan potentiaalinen menetelmä partikkelikoon pienentämiseen. Muita menetelmiä, kuten NTA-analyysia (engl. Nanoparticle Tracking Analysis), tarvitaan kuitenkin vahvistamaan muodostuneiden lipidi-DNA-partikkeleiden koosta saatuja tuloksia.

MDMA is an illegal stimulant known for its empathy-enhancing effects. Its positive effects are mainly based on increasing the concentrations of monoamines such as serotonin (5-HT), dopamine (DA) and norepinephrine (NE). In addition to its positive effects, MDMA can cause adverse effects such as hyperthermia and neurotoxicity. Especially with long-term use, MDMA can cause serotonergic and dopaminergic neurotoxicity. In addition, there are also indications of MDMA-induced neurotoxicity in systems where gamma-aminobutyric acid (GABA) functions as the main neurotransmitter. Glutamate decarboxylase (GAD) 67 is an enzyme that synthesizes GABA from glutamate and is a specific marker for GABAergic cells. The amygdala is a nucleus in the brain that regulates anxiety and fear response. In addition to GABAergic interneurons, there are also glutamatergic cells in the basolateral nucleus (BLA) of the amygdala, however in the central nucleus (CeA) there are only GABAergic cells. Disturbances in the GABAergic system can predispose to psychiatric diseases such as anxiety. The aim of thisstudy was to investigate the effects of MDMA (20 mg/kg) on the number of GAD67-positive cells in two nuclei of the mouse amygdala, BLA and CeA. In addition, this study aimed to examine the importance of the dose (4 or 16 injections) for neurotoxicity and the duration of the effects (2, 7 or 30 days). Adolescent wild type mice were divided into 12 groups according to the treatment (MDMA or saline), dose and timepoint. After euthanasia, the brain sections at the level of the amygdala were collected and stained with an immunohistochemical method and imaged using a confocal microscope. This study showed that MDMA reduced the number of GAD67-positive cells in the BLA when mice were given a total of 4 injections. This effect lasted up to 30 days. In contrast, MDMA did not reduce the number of GAD67-positive cells in the BLA in mice that were given 16 injections. Also, MDMA did not decrease the number of GAD67-positive cells in the CeA, regardless of dose. Statistical significance could have been improved, for example, by using more mice or analysing more sections from each individual animal. It is important to continue studying the effects of MDMA to better treat and prevent its adverse effects. In addition, increased understanding would urge users to exercise caution when using MDMA.

The package leaflet (“leaflet”) is a technical document included in medicine packages to provide information about the medicinal product to the user. With the EU now encouraging the adoption of eHealth, it can be assumed that written medicine information would be included in the digitalisation process. Medicine users’ views on electronic forms of medicine information should be assessed before any changes can be made, but so far there is very little data on this. The aim of this study was to find out what kind of leaflet medicine users would prefer and how they would feel about an electronic leaflet. The main aim was to find out if there is a difference in preferences between different types of medicine users and between medicine users of different ages in the provision of a package leaflet. The study also sought to find out if the current leaflet is being read by medicine users. This study was conducted by carrying out a survey to pharmacy customers over the age of 16 collecting prescription medication(s) for themselves (n = 110). The data was collected at one retail pharmacy in Helsinki, Finland during July 2020. The data was analysed quantitatively. This study found that medicine users generally feel positively about an electronic leaflet (liked by 63%) and many are open to idea of an electronic leaflet (75%). The majority (88%) could see positives in using an electronic leaflet, regardless of leaflet preferences. The study did not find a difference between new and repeat medicine users in the preference for a particular leaflet format, but age is correlated with the preference for a particular leaflet type, with younger medicine users wanting an electronic leaflet as often as older medicine users want a printed leaflet. Having the leaflet appear in My Kanta pages after the medication has been dispensed was found to be the most popular way to receive an electronic leaflet. This study also found that there is a difference between new and repeat medicine users when it comes to reading the leaflet after a medication has been dispensed. With the current printed leaflet 81% of repeat medicine users and 38% of new medicine users do not read it. The most common reason given for not reading the leaflet was that the participant had read it before and did not feel the need to read it again. According to this study, medicine users, especially younger medicine users, feel positively about the idea of an electronic leaflet, which is encouraging for the future of an electronic leaflet. The results are in line with prior research, but also suggest that more medicine users feel positively about the idea of an electronic leaflet than before. The leaflet reading behaviours of medicine users also highlight the need for a system, where a medicine user can be alerted to any changes in the leaflet, which is something only an electronic system could do.

Heart failure is a global health issue that can result from various factors, one of which is myocardial infarction. The adult human heart has limited regenerative capacity to cover the loss of cardiomyocytes after myocardial infarction with new cardiomyocytes. The main responses to the loss of cardiomyocytes are fibrotic scar formation and the hypertrophy of remaining cardiomyocytes. Prolonged hypertrophy eventually leads to heart failure. Current treatments for heart failure only relieve the symptoms. Inducing cardiac regeneration could be one possible way to prevent and treat heart failure. Thus, to develop medical treatments that enhance the regenerative capacity, a comprehensive understanding of precise cellular mechanisms behind heart regeneration is crucial. The objective of this study was to establish a high-content analysis method for human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) utilizing the Cell Painting assay to identify and categorize morphological alterations induced by various compounds in hiPSC-CMs. To evaluate the morphological impacts, dozens or even hundreds of cell features were measured at the same time. hiPSC-CMs were exposed to two hypertrophy inducers, endothelin-1 and angiotensin II, and to doxorubicin, which is known to be a cardiotoxic compound. In addition, the effects of a GATA4- targeting compound, C-2021, on hiPSC-CMs were examined. C-2021, was expected to have antihypertrophic effect on the cells. Previously used methods, proBNP staining and qPCR, were used to validate the novel method. According to proBNP staining and qPCR, endothelin-1 induced cardiomyocyte hypertrophy greater than angiotensin II. Compound C-2021 did not show statistically significant antihypertrophic properties after hypertrophic stimuli, but some tendency the alleviate the hypertrophy was noticed. Moreover, by utilizing different data processing programs a novel analysis method was developed. With this method, the different treatment groups were clustered based on the morphological alterations caused by compounds exposures. The hiPSC-CMs exposed to endothelin-1, angiotensin II or doxorubicin showed a different morphological profile compared to the control group hiPSC-CMs. Compound C-2021 was also observed to affect cell morphology. However, the data analysis still needs improvements in order to detect which cell features these compounds affect.

Objectives To investigate distribution and causes of drug shortages in five selected countries with data from public shortage notification registers. Design Statistical retrospective analysis of national drug shortage registers Data Public shortage notification register data from the first nine months of 2020 in Finland, Sweden, Norway, Spain, and the United States, partly combined with national drug registers. Results Altogether 5132 shortage reports from Finland (n=1522), Sweden (n=890), Norway (n=800), Spain (n=814), and the United States (n=1106) published during the first nine months of 2020 were found in the study. More than half (54%) of the active ingredient level shortages (classified by ATC code) found occurred in only one country, and only 1% occurred in all five countries. On a country level, 19-41% of the shortages were found only in a given country. The distribution of shortages by ATC category and drug form was significantly different between countries, especially between the US and European countries. Injectables were found to have an especially high shortage risk in the US, much less so in European countries. On the other hand, some drug classes were in shortage almost exclusively in Europe but not in the US. Conclusions Although drug shortages are a growing global problem, drug shortages are rarely global, but typically occur only in some countries, while other countries have an uninterrupted supply of the same drug. Drug shortages should be seen not just as a problem of manufacturing disruptions, but as a question of equitable and effective distribution of drug supply on an international level. Price differences and other commercial issues could be factors behind variation of shortages found between countries.Considering the limited and probing nature of the study, further research of shortage register data is certainly warranted. International comparative register study is a meaningful and valid method for further understanding in this field.

The OATP1B3 belongs to the organic anion transporting polypeptides (OATPs) encoded by the SLCO (solute carrier organic anion) genes which belongs to the SLC (solute carriers) gene superfamily. It is an influx transporter which is primarily expressed on the basolateral membrane of the hepatocytes. It transports many endogenous substrates as well as clinically important drugs such as thyroid hormones and statins into hepatocytes and thus participates in the first step of hepatic metabolism. Single nucleotide polymorphisms (SNPs) of the SLCO1B3 gene can affect the pharmacokinetics and pharmacodynamics of its substrates. The aim of this study was to set up and optimize an in vitro method to study the function and expression of the OATP1B3 transporter and its genetic variants. SNPs 334T> G (Ser112Ala), 699G> A (Met233Ile) and 767G> C (Gly256Ala) and stop codon TAA were introduced into the SLCO1B3 gene by site-directed mutagenesis. Recombinant baculovirus vectors containing the genetic information of OATP1B3 and its variants were used to transiently transfect the HEK293 cells. After optimizing the substrate incubation time and concentration, as well as the viral load and selecting the fluorescent substrate (8-FcA), the uptake assay was used to determine the transport activity of the OATP1B3 variants in HEK293 cells. The transport activity of the artificial WTP variant was also investigated in this study. The transport activities of the Ser112Ala, Met233Ile and Gly256Ala variants did not change significantly from the wild type although the transport activity of the Met233Ile variant appears to be slightly impaired. In turn the WTP variant was unable to transport 8-FcA. Based on this study the function of OATP1B3 variants can be studied using recombinant baculovirus to transiently transfect the HEK293 cells. 8-FcA can be used as a probe substrate in these studies. The results of this study confirm previous knowledge of the functioning of Ser112Ala, Met233Ile and Gly256Ala variants. More studies are needed about the effects of these variants on the transport of OATP1B3 drug substrates. Also studies about the location, cell membrane and total cell expression of the WTP variant are needed to evaluate reliably the reasons of its inactivity.

Evidence based medicines alongside with age-appropriate dosage forms are essential in enabling appropriate treatment for any patient group. Pediatric pharmacotherapy, however, is lacking age-appropriate dosage forms and research-based evidence regarding the dosing, efficacy, and safety of medicines. Orally administered drugs require manipulation to enable administration and are often used against the indications approved in the marketing authorization and summary of product characteristics (SmPC). This off-label use puts pediatric patients at risk for medicational errors and adverse drug reactions. The aim of this study was to investigate recent trends in oral dosage forms used in pediatric randomized controlled trials (RCTs), with emphasis on age appropriateness. The results could be utilized in developing evidence-based dosage forms, suitable for all pediatric patients aged 0-17 years, and manufacturable in a small scale in a hospital pharmacy. This study was conducted as a systematic review following the PRISMA Statement. The literature search was carried out from Pubmed and Scopus databases and it covered a five-year period of 2015-2020. References from the databases were entered to the Covidence systematic review platform. After removing duplicates 3333 articles were left for screening. Two independent researchers selected the articles first screening by title and abstract, and then by full text review. A qualitative content analysis was conducted on the included articles. Altogether 77 articles met the inclusion criteria. Dosage forms included were tablets (n=37), liquids (n=21), capsules (n=18) and multiparticulates (n=6). Majority of the dosage forms were conventional (n=49) compared to more advanced novel modified release and fixed-dose combination formulations (n=33). Based on our results, orally administered dosage forms used in the recent pediatric RCTs are still limited by poor acceptability, palatability, and the need to manipulate dosage forms to enable administration. These issues are similar to the ones related to the off-label use of medicine that compromise patient safety. Majority of the dosage forms included in our study were tablets, indicating a positive shift towards more safe and acceptable dosage form. Formulations were also evolving towards dispersible, extended-release and fixed-dose combinations that offer additional benefits for pediatric patients. The low number of children < 2 years old included in study populations and the poor acceptability profile reported by most studies limit our conclusions on an ideal age-appropriate dosage form. Further research is needed on unifying the guidelines used in pediatric drug development.