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In medicines APIs are most oftenly at solid form. Crystal forms are more stable than amorphic solid form. Crystals are hold together by intermolecular interactions. Strongest and most common intermolecular interaction in crystals is hydrogen bond. Crystallisation is affected by thermodynamics and kinetics. Same phenomena effect also dissolving of crystals. New APIs often have a poor water solubility which makes them difficult to use. Cocrystals are one way to improve physical characteristics of molecules and most of all solubility. In co-crystals two different solid molecules are crystalliced in a same crystall lattice. Itraconatzole is an API with a poor water solubility. Itraconatzole can form cocrystals with many bicarbocsylicacids. The smallest bicarbocsylicacid that had formed co-crystal with itraconazole is malonic acid. The purpose of the experiment was to grow itraconazole malonic acid co-crystal, which is big enough for single crystall x-ray diffraktion. With SXRD it is possible to find out how molecyles are placed in a crystall lattice. For SXRD the single crystal is not allowed to have a single mistake in its lattice. Itraconazole and malonic acid were dissolved to 1,2-dichloroethane-2-butanone and tetrahydrofurane-chloroform for growing up a single crystal. Crystallisation methods used were evaporation of solvent, adding antisolvent and cooling down of solution. Formed crystalls were analysed with DSC, raman, XRD and TGA. It was succeeded to crystallise itraconazole malonic acid co-crystals with used methods. The formed crystalls were needlelike and packed in small drifts. Any crystall big enough for SXRD was not succeeded to grow up with the methods used. Growing up a bigger co-crystal needs the use of new methods or optimation of the ones used in this experiment.

The pet medication industry is growing but there are still challenges especially in feline medication. Palatable flavours, efficient taste masking technologies and easily administrable dosage forms are needed to facilitate feline medication. Based on the literature review, there is only little information about cat's preference to individual flavours. The methods for palatability testing should be improved to achieve reliable results. Most common taste masking technologies are flavouring and tablet coating. In experimental section different flavours for taste masking were studied. Five main flavours were selected: phenylalanine, leucine and methionine as possibly good flavours and arginine and denatonium benzoate as bad flavours. In preformulation experiments tableting characteristics, thermal behaviour and crystal structure of flavours were analysed. The aim was also to study their possible incompatibilities with tablet excipients. The main compatilibility study method was X-ray powder diffraction (XRPD), but differential scanning calorimetry (DSC) was also used. Excipient povidone (PVP) was incompatible with nearly all of the main flavours. The use of lactose as an excipient was excluded because of the risk of the Maillard reaction. In tableting studies a tablet mass containing microcrystalline cellulose (MCC), calcium hydrogen phosphate dihydrate, mannitol, hydroxypropyl cellulose (HPC), crospovidone, talc and sodium stearyl fumarate was produced. Minitablets of diameter 3 mm without any flavours were compressed. Also minitablets with flavours phenylalanine and denatonium benzoate were compressed. Minitablets complied with the European Pharmacopoeia tests for uniformity of mass, disintegration and friability. However, characterization and handling of minitablets was found to be challenging due to very small size of the tablets. Minitablets are a promising technology for facilitating feline medication in the future.

Voltage-gated sodium channels play an essential role in the function of the nervous system as they are responsible for producing action potentials. Abnormal activity of sodium channels is in connection to several diseases such as epilepsy and chronic pain. Voltage-gated sodium channel blockers which are selective towards a specific isoform could provide more efficient and better tolerated drugs to treat these diseases when compared to the drugs used today. Clathrodin is an alkaloid isolated from Caribbean sea sponge Agelas clathrodes. Bioactivity studies have shown that clathrodin changes the function of voltage-gated sodium channels. The aim of this study was to synthesize two kinds of structure analogs of clathrodin and study their structure-activity relationship towards different isoforms of voltage-gated sodium channels. The study is part of the MAREX project (Exploring Marine Resources for Bioactive Compounds: From Discovery to Sustainable Production and Industrial Applications) funded by the European Union. Intention of the project is to find new bioactive compounds in marine organisms. A four-step route was developed for synthesizing 2-aminobenzothiazole analogs. A three-step route was developed as well but the last step seemed to be problematic for some of the compounds. The three-step route provided new compounds as intermediates and some of them were sent to tests for activity. Synthesis of 1H-pyrrole-2-carboxamide analogs of clathrodin failed. 4,5-dihydrooxazole was recognized as a problem as it was formed as a result of a cyclization reaction when bromination was tried on the intermediate. The formed structure was used in synthesizing 2-(1H-pyrrol-2-yl)-4,5-dihydrooxazole analogs of clathrodin. These reactions failed to give any final products which could have been tested for activity. Eight synthesized compounds were sent to tests for activity. Results were received from two of them and they showed no activity towards the voltage-gated sodium channels in 10 µM concentrations. Discussion about structure-activity relationship is not possible based on two compounds only.

During the past few decades the focus of the pharmacy profession has shifted from medicinal products towards ensuring the welfare of the patient. The concept of pharmaceutical care emphasizes that the role of the pharmacist is to ensure the quality and safety of pharmaceutical therapy in collaboration with the patient. The concept of clinical pharmacy, on the other hand, highlights that the pharmacist should take the responsibility of the efficiency, safety and cost efficiency of the patient's pharmaceutical treatment together with other health-care professionals. Patient centered pharmacy services have been increased in Finnish hospitals and health care-centers during the last 10 years, for example, in the form of pharmacy services provided on the wards. Previous studies have shown that Finnish hospital pharmacists want to develop and increase clinical pharmacy services in hospitals but feel they are not competent enough to manage them. To develop and increase the number of clinical pharmacy services it is important to ensure the pharmacy professionals have support to their continuing professional development. The General Level Framework (GLF) has been developed in the UK to support the professional development of pharmacy professionals and its value has been shown in various studies. The aim of this study was to give information of the state of Finnish hospital pharmacy and its development needs as perceived by Finnish hospital pharmacists, and to investigate how the GLF can be utilized in Finnish hospital pharmacy. The study was conducted using two different research methods: a semi-structured interview was designed to investigate hospital pharmacists' perceptions of hospital pharmacy, it's development needs and utilization of the GLF; in addition, hospital pharmacists selfassessed their clinical pharmacy related competencies using the GLF. All of the participants of the study were participants of a clinical pharmacy course by University of Helsinki. In total 11 hospital pharmacists took part in the interview. Also 41 pharmacists self-assessed their competencies in clinical pharmacy using the GLF: eight of them completed the self-assessment twice with a six month period between the assessments. The interviewed pharmacists felt that the pharmacy curriculum should focus more on the skills and competencies needed in hospital pharmacy. On the other hand, they felt that the tasks of hospital pharmacists did not necessarily allow them to use their actual knowledge of pharmacy. They perceived that the future of hospital pharmacy lies in services of clinical pharmacy, although they felt that they were not competent enough to manage them. The GLF self-assessment showed that the clinical competencies of the participants were average, and there was no change in the competence of the participants during the six month period. However, the interviewed pharmacists felt that the GLF can be used as a tool for support the hospital pharmacists' professional development and continuing professional development in clinical pharmacy. They also perceived that there is a need for further studies on the clinical pharmacy services and their benefits, and that the task distribution of health care professionals in hospitals must be re-considered. With these actions, the challenges hindering the development and increase of clinical pharmacy services in Finnish hospitals could be overcome. They described that the greatest challenges to overcome were prejudices against pharmacist working on the wards and a lack of resources. In order to develop and increase clinical pharmacy services in Finnish hospitals and other health-care organizations further reseacrh on the benefits of clinical pharmacy should be conducted. It must also be ensured that all health-care professionals and decision makers are aware of the studies already made about clinical pharmacy and its benefits. The skill-mix of health care professionals taking part in a patient's treatment must be reconsidered in order to ensure that the patient receives the best, most efficient and safest possible medicinal care. The GLF can be used as a tool to define the role of a clinical pharmacist in Finland. There is a need for more clinical pharmacy education so that Finnish pharmacists can feel competent enough to manage clinical pharmacy tasks. The GLF can be used as a tool to support the professional development and continuing professional development also in Finland.

In cancer therapy nanocarriers can be loaded with therapeutic or diagnostic agent and nucleic acid sequences. Targeting moieties can be attached to the nanocarrier for passive or active targeting or carrier can be labeled with radioactive isotope for imaging or radiotherapeutic purposes. Enclosing the drug in a nanocarrier may improve the molecule's physico-chemical properties, bioavailability, reduce side-effects, longer the circulation time and dosing interval, and improve uptake in the target tissues. Thus, the efficacy of chemo- or radiotherapeutic could be improved. It may lead to improved survival. Pro gradu investigates nanocarriers' role in cancer therapy. Regardless of research, continued for decades, only 2 (Europe) or 3 (United States) nanoparticle formulations are approved in cancer therapy. Major limiations are inefficient uptake in the target tissue, immunogenicity of nanoparticles and targeting ligands, and lability. The aim of this study was to investigate pre-targeting of 99mTc-labeled, PEGylated and biotinylated liposomes into human ovarian adenocarcinoma cells in vitro and in mice in vivo. Targeting moiety used was biotinylated cetuximab (Erbitux®), an antibody that binds into EGF-receptors, over-expressed in these cells. Pre-targeting was compared to active one step-targeting, with antibody attached to liposomes, and passive targeting. Development of more accurate imaging techniques has accelerated the investigation of targeted nanoparticles. Molecular imaging enables real-time tracking of nanoparticle distribution and metabolic changes. In literature review, SPECT and PET imaging in cancer therapy and nanoparticle research, will be discussed. These imaging methods overcome challenges in sensitivity and accuracy, faced by other imaging methods. In this study we also investigated the biodistribution of 99mTc-labeled liposomes in mice using NanoSPECT-CT-device. Activity in tumor, spleen and liver was quantified using InVivoScope-software and gamma counter and these results were compared. In in vitro study, pre-targeting method was 2,7and 3,5-times more efficient compared to the liposome controls in SKOV3 and SKOV3.ip1 cell lines, respectively. Although, one-step targeting formulation targeted the cells even better. In in vivo -study, i.p.-administered liposomes distributed into tumor more efficiently compared to i.v.-administered liposomes. I.p. pre-targeting method was 1,24-fold more efficient compared to passive targeting, considering the % ID / g tissue. However, %ID/organ in pre-targeting method was 5,9 % whereas passive targeting reached the value of 5,4 %. Conclusively, the difference between pre-targeting and passive targeting was modest. InVivoScope and gamma counter quantification results didn't correlate. Further investigation is needed and protocol optimization required in targetin liposomes into tumors.

Obesity is a health problem linked to Western lifestyle and it is becoming more general. The complexity of regulation of eating makes it difficult to regulate body weight, when multiple neural networks and regions of brain have overlapping functions regarding to energy gain. Sufficient amount of energy is vital for individuals surviving in their living environment. Cholinergic messaging in brain is wide and for example nicotine is known for its appetite reducing effect. Anorexigenic proopiomelanocortin neurons mediate the effect of nicotine. In nucleus accumbens and central nucleus of amygdala extracellular levels of acetylcholine rise during meal, which promotes satiety. Satiety inhibits eating behavior between meals. Amygdala is a part of limbic system and in earlier knowledge it was associated only to regulation of memory, conditioning and fear. Nowadays importance of amygdala in eating behavior research is rising, but most of the studies focus on the effect of cue in regulation of eating. Cholinergic messaging is vigorous in the amygdala and is received from opposite areas of brain between basolateral and central amygdala and therefore this master’s thesis examined the effect of cholinergic messaging in amygdala on regulation of eating behavior. C57BL/6JRcc male mice were stereotaxically implanted with guide cannulas either in the basolateral complex of amygdala (n=10) or central nucleus (n=13). After recovering and habituation to automated pellet dispenser mice were treated with nicotinic and muscarinic receptor agonists and antagonists and eating behavior was recorded for six hours. Nicotine, administered to central and basolateral part of amygdala, lowered the number of pellets mice ate. In central nucleus effect was dose dependent. Mecamylamine had time related effect on eating behavior in basolateral amygdala, but dose dependent response was seen only in cumulative results. Oxotremorine was the only compound which created statistically significant interaction between time and dose. Result was seen in both groups. Scopolamine reduced eating behavior in central nucleus and dose dependency was seen. In basolateral complex scopolamine had time related effect, similar to mecamylamine. The results suggest that amygdala regulates eating behavior even without cue.

Some problems in dry powder inhaler formulation include low dose efficiency and changes in dispersibility during storage. For lung deposition particles should have aerodynamic size of 1 - 5µm. Poor dispersion of drug particles from carriers' surface is thought to be the main reason for low dose efficacy. A tertiary component of small particles has been generally added to formulation to improve fine particle dose. Small particles are usually manufactured by micronization. This may induce crystal defects and amorphous sites on the surface of crystals. Amorphous sites are metastable and they may crystallize during storing. Changes in particles crystallinity may have an action on efficiency and stability of dry powder inhalers. Conditioning is designated as stabilisation of particles surface by mixture of solvent vapour and inert gas. Vapour may also dissolve surface roughness. This is called deliquescence. Ostwald ripening is phenomenon whereby small particles dissolves and recrystallizes onto larger crystals. This can be extended for surface asperities. Amorphous materials have also better solubility than crystalline materials so amorphous sites may also dissolve and recrystallize onto crystalline surface. Amorphous sites may crystallize spontaneously by absorbing plasticizing agents from vapour phase or by influence of temperature. The purpose of this work was to study process variables in conditioning and their effect on modification of surface roughness and stabilization of micronized α-lactose monohydrate and test drug substance. The purpose was also to study how surface modification and stabilization effects on powders flowability and stability of dry powder inhaler. The dry powder inhaler contained two different vicinity of lactose and two different drug substances. Conditioning was based on evaporation of liquid from open surface. Studied process variables were temperature of powder, temperature of bath of liquid phase and flow rate of nitrogen gas. The aim of this study was to form a process design for conditioning of new substances, to improve powders flowability and to remove changes in fine particle dose during storage. Surface roughness was studied by laser diffraction analysis and specific surface area measurements and also by electron microscopy. Specific surface area was measured by nitrogen adsorption method. Stabilization of amorphous sites ware studied by dynamic vapour sorption. Flowability was measured by angle of repose and with FlowPro device. Fine particle dose was measured with next generator impactor device. The study showed that increasing the amount of solvent in vapour increases surface smoothness and stabilization. Also increase of temperature of sample increased stabilization. Influence of temperature on surface smoothness was not as clear. Changes in temperature may have altered adsorption and kinetic of crystallization of dissolved molecules. Flowability of lactose was significantly improved. Condition did not improve dry powder inhalers fine particle dose, but there was significant difference between different process conditions. This was concluded to be caused of surface modification. It was also shown that different process conditions affected on formulations stability.

Raman spectroscopy is based on vibrations that occur between the atoms of a compound. The overall structural energy is derived from the electronical energy as well as vibrational, rotational and translational energy. In Raman spectroscopy the vibrational and rotational energies are essential. Usually the excitation energy used in Raman spectroscopy can be either in the region of visible light or NIR. The sample absorbs the energy and energy is also scattered back to all possible directions. Elastic scattering is called the Rayleigh scattering. In that case the back-scattered photons have an equal energy as the original excitation energy. However, some of the scattering happens inelastically and it forms the basis of Raman-phenomena. If the detected photons have smaller energy than the original, it is called the Stokes scattering. If the energy is bigger, it is anti-Stokes scattering. Raman is typically very rare and weak phenomenon. The spectral features in Raman spectra consist of the intensities and energies of the back scattered photons. Raman spectroscopy provides very accurate and detailed structural information on the molecule. It is basically a label-free technique with minimal need for sample preparation and the measurements can also be carried out e.g. through container walls. Further, Raman is quite insensitive to hydrous samples and it is suitable to solutions and biological assessments. However, there are some drawbacks that are formed by the luminescence phenomena i.e. fluorescence. Strong fluorescent backgrounds can mask the relevant Raman features in spectra because Raman and fluorescence are competetive processes. For instance many drug molecules have such structures that they cause strong fluorescence. It is also one of the reasons that pharmaceutical applications and measurements have been partly limited due to this problem. There are applications to improve and enhance a Raman signal. For example resonance phenomena and SERS are favored. To solve the fluorescence-related problems there are also means; one can change the laser wavelenght, photobleach the sample or apply different kinds of data manipulation techniques to the spectral data achieved. There are drawbacks with these methods. They can be slow, complex, damage the samples and still insufficient fluorescence suppression is a problem. In this study a novel time-gated CMOS-SPAD detection technique is applied to non-fluorescent and fluorescent drug measurements. The new detection system has a programmable on-chip delay time and it is synchronized with a picosecond pulsed laser. The scattered photons can be measured in the time scale when they are simultaneously measured in traditional energy and intensity wise. Raman scattering occurs in the timescale of sub-picoseconds while the fluorescence phenomena happen typically in the order of nanoseconds. This time difference can be exploited effectively to suppress the fluorescence. In the literature review of this study the basis of vibrational spectroscopy is introduced - especially Raman spectroscopy. The techniques related, as well as the novel time-resolved technique are covered. Further, different kinds of applications in the field of Raman spectroscopy are reviewed, mainly pharmaceutics-related and biologically relevant applications. In the experimental work the focus was to compare a continuous-wave 785 nm laser setup coupled with the CCD-detector to the pulsed picosecond 523 nm laser coupled with the CMOS-SPAD-detector. The measurements were performed on different kinds of drugs, both non-fluorescent and fluorescent. The aim was to obtain information on the effectiveness of CMOS-SPAD-technique on fluorescence suppression for solid drugs and solutions. Secondary goals were to collect knowledge on the similarities and differences between the Raman setups used for solution measurements, to optimize and discuss the key elements of setups for solids and solutions and to show preliminarily the applicability of the CMOS-SPAD-system on fluorescent drug's solutions as well as find out the requirements related to quantitative assessments using Raman spectroscopy. In drug research there is also constant need for reliable in vitro cell assays. The assessments made in this study may prove useful to the future applications e.g. measurements with living cells. An effective fluorescence suppression was achieved to strong fluorescent backgrounds using the novel time-resolved CMOS-SPAD-detection system coupled with the pulsed picosecond 532 nm laser. The setup is potentially a convenient tool to overcome many fluorescence-related limitations of Raman spectroscopy for laboratory and process analytical technology (PAT) use in the pharmaceutical setting. The results achieved encourage to consider that with careful calibration and method validation there is potential for quantitative analysis, biopharmaceutical and biological applications e.g. in vitro cell studies where most Raman techniques suffer from strong fluorescence backgrounds. Other potential fields for future applications can be also considered.

Cortisol is a vital hormone for normal bodily functions. Both physical and mental stress, as well as many diseases like the Cushing syndrome are known to increase the human cortisol levels. These levels can be measured in many biological matrixes, such as saliva. Traditionally, these measurements have been done by using immunoassays or liquid chromatographic-mass spectrometric methods (LC-MS). However, in the last few years, ambient ionization techniques, which are quick and easy to use, have also proven suitable for quantitative analysis of compounds in biological matrixes. Thus, these techniques could offer an alternative to traditional methods in the analysis of cortisol from human saliva. The aim of this study was to investigate the suitability of desorption atmospheric pressure photoionization (DAPPI) for quantitative analysis of steroids in saliva. The investigated steroids were dehydroepiandrosterone (DHEA), cortisol and testosterone. Because of the low quantities of testosterone and DHEA in saliva, the study was mainly focused on cortisol analysis. In this study, the ionization mechanism for the steroids was observed to be proton transfer with every tested spray solvent (acetone, chlorobentzene and toluene). Even though the choice of spray solvent did not change the ionization mechanism, it affected the efficiency of ionization. In cortisol measurements acetone was observed to be the best solvent. The temperature of the microchip, as well as the UV-lamp used (dc- or rf-lamp), only affected the ionization slightly. In this study, measuring cortisol in non-pretreated saliva was not successful. However, solid phase extraction (SPE) method for the pretreatment of saliva was optimized with high recovery for cortisol (106 %). The detection limit for cortisol (50 nM) in water samples and the linear area of cortisol in both water and pretreated saliva samples (500 nM - 10 µM) were also determined. Poor repeatability of DAPPI-system was the main challenge in these measurements. The DAPPI-MS-method developed in this study is suitable for analyzing cortisol in pretreated saliva samples. However, without further development it is not sensitive enough to be used in quantitative analysis of cortisol in salivary levels.

COVID-19 pandemic has caused a global crisis and its effects have also been reflected to pharmaceutical supply in Finland. At the beginning of the crisis the effects were especially evident in the consumption of self-medication analgesics, prescription drugs and drugs related to respiratory diseases. In a global crisis, collaboration between the public, private and third sector is becoming increasingly important, and it is important to consider how to develop the capacity for collaboration between organizations in different sectors during a pandemic. The purpose of this study was to find out how the cross-sector collaboration between the public, private and third sector of the pharmaceutical supply in Finland was organized in the crisis caused by the COVID-19 pandemic, what was the role of the cross-sector collaboration and how the preparedness and crisis management of the drug supply could be improved. The study was conducted as a semi-structured interview survey and the interviewees were selected to cover the various sectors of Finnish pharmaceutical care as well as possible. The analysis was performed by the Gioia method and thematic design. Based on the study the organization of cross-sector collaboration was both operator- and authority-oriented and the legislation and environment in the drug supply created the framework for the crisis management. Both the authorities and the advocacy organizations can be described as having acted as hubs for organization. There was no clear crisis organization in drug supply, but different actors were involved in the crisis management at different stages of the crisis. The role of collaboration was emphasized in the sharing of information and resources and in joint solution of problems. The collaboration enabled foresight and preparedness, a focus on core tasks and crisis management, and mutual benefit. Lessons learned from the COVID-19 pandemic include the need to increase and intensify collaboration, increase crisis plans and crisis training, update the system of security of supply and mandatory reserve supplies, increase self-sufficiency, and increase overall governance. Cross-sectoral collaboration was seen as useful in crisis management of the crisis in the drug supply chain. The collaboration promotes the formation of a common picture of the situation and the flow of information from the field to decision-makers. Comparing the results of this study with the literature it can be said that the results partially support the previous literature. However, crisis management of the pharmaceutical supply chain from the organization of cross-sectoral collaboration point of view has not been studied in the past.