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Medication adherence is a key precondition of the effectiveness of evidence-based therapies. However, in real-life settings, non-adherence to medication is still very common. This leads to profound health and economic consequences at both individual and public health levels. The problem of non-adherence has been extensively studied in the last 50 years. Unfortunately, with more than 130,000 scientific papers published on that subject so far, we are still far from finding an ultimate solution. This is, at least partly, due to fragmented and poor-quality research that has been conducted in this field sometimes. To overcome this deadlock, there is a need to stimulate the adoption of best practices in medication adherence-related research in a systematic way. Therefore, herein we propose the establishment of dedicated medication adherence research Centres of Excellence (CoEs). These Centres could not only conduct research but could also create a profound societal impact, directly serving the needs of patients, healthcare providers, systems and economies. Additionally, they could play a role as local advocates for good practices and education. In this paper, we propose some practical steps that might be taken in order to establish such CoEs. We describe two success stories, i.e., Dutch and Polish Medication Adherence Research CoEs. The COST Action “European Network to Advance Best practices & technoLogy on medication adherencE” (ENABLE) aims to develop a detailed definition of the Medication Adherence Research CoE in the form of a list of minimal requirements regarding their objectives, structure and activities. We hope that it will help to create a critical mass and catalyse the setup of regional and national Medication Adherence Research CoEs in the near future. This, in turn, may not only increase the quality of the research but also raise the awareness of non-adherence and promote the adoption of the best medication adherence-enhancing interventions.

Pulmonary drug delivery has been used for decades to treat local diseases like asthma. When using the pulmonary route to deliver drugs, several important lung features are being used, such as a large surface area available for absorption, high organ vascularization, and a thin blood-alveolar barrier. Pulmonary drug delivery systems on the market are formulations with a rapid release, which leads to a high drug concentration initially and a prompt decline in concentration shortly thereafter. This could cause unfavourable adverse effects or toxicity to the lung tissue at the onset of the release and could also result in decreased efficacy. To overcome these challenges, there is a need to develop controlled release drug delivery systems to improve the therapeutic effectiveness of inhaled drugs. When a drug is inhaled, the drug particles will deposit in the lung, and the drug needs to dissolve in the lung fluids before the drug is available for uptake locally or in the systemic circulation. The absorption inhaled drug thus depends on the dissolution of the drug particles in the lung fluid. As a result, it could be possible to prolong the duration of the drug effect, by prolonging the time it takes for the dissolution of the drug particles. Due to this, in vitro methods analysing the dissolution of the drug particles in the lung are of high relevance for the development of novel pulmonary drug delivery systems. It is therefore of high importance that the dissolution profiles that are measured are well understood. The overall aim of this thesis was to evaluate and characterise an in vitro dissolution method (Transwell system) for assessment of novel pulmonary drug delivery systems, with a focus on future controlled release systems. A developed mechanistic model was used to analyse experimental dissolution data and to predict which process was the rate limiting step in the obtained profiles. The developed mechanistic model provided the same rank order as the Weibull fit, however the model provided additional detailed understanding of the used dissolution process and setup. In addition, two novel controlled release drug delivery systems, mesoporous silica particles and hyaluronic based hydrogels, were successfully analysed using this in vitro dissolution system. Both delivery systems showed a promising aerosolization and control over the release profiles. Finally, the micellar contribution to diffusion of poorly soluble inhaled drugs during in vitro dissolution was defined and validated using the obtained in vitro dissolution profiles. Physiologically based biopharmaceutics modelling tools were successfully established for Bud, BDP and FP using the diffusivity values taking into account the micellar contribution of the surfactant.

Colonic mucus plays a key role in colonic drug absorption. Mucus permeation assays could therefore provide useful insights and support rational formulation development in the early stages of drug development. However, the collection of native colonic mucus from animal sources is labor-intensive, does not yield amounts that allow for routine experimentation, and raises ethical concerns. In the present study, we developed an in vitro porcine artificial colonic mucus model based on the characterization of native colonic mucus. The structural properties of the artificial colonic mucus were validated against the native secretion for their ability to capture key diffusion patterns of macromolecules in native mucus. Moreover, the artificial colonic mucus could be stored under common laboratory conditions, without compromising its barrier properties. In conclusion, the porcine artificial colonic mucus model can be considered a biorelevant way to study the diffusion behavior of drug candidates in colonic mucus. It is a cost-efficient screening tool easily incorporated into the early stages of drug development and it contributes to the implementation of the 3Rs (refinement, reduction, and replacement of animals) in the drug development process.

The subtilisin-like macrocyclase PatGmac is produced by the marine cyanobacterium Prochloron didemni. This enzyme is involved in the last step of the biosynthesis of patellamides, a cyanobactin type of ribosomally expressed and post-translationally modified cyclic peptides. PatGmac recognizes, cleaves, and cyclizes precursor peptides after a specific recognition motif comprised of a C -terminal tail with the sequence motif-AYDG. The result is the native macrocyclic patellamide, which has eight amino acid residues. Macrocyclase activity can be exploited by incorporating that motif in other short linear peptide precursors, which then are formed into head-to-tail cyclized peptides. Here, we explore the possibility of using PatGmac in the cyclization of peptides larger than the patellamides, namely, the PawS-derived peptide sunflower trypsin inhibitor-1 (SFTI-1) and the cyclotide kalata B1. These peptides fall under two distinct families of disulfide constrained macrocyclic plant peptides. They are both implicated as scaffolds for drug design due to their structures and unusual stability. We show that PatGmac can be used to efficiently cyclize the 14 amino acid residue long SFTI-1, but less so the 29 amino acid residue long kalata B1.

In this work, simulations of intestinal peristalsis are performed to investigate the intraluminal transport of macromolecules (MMs) and permeation enhancers (PEs). Properties of insulin and sodium caprate (C10) are used to represent the general class of MM and PE molecules. Nuclear magnetic resonance spectroscopy was used to obtain the diffusivity of C10, and coarse-grain molecular dynamics simulations were carried out to estimate the concentration-dependent diffusivity of C10. A segment of the small intestine with the length of 29.75 cm was modeled. Peristaltic speed, pocket size, release location, and occlusion ratio of the peristaltic wave were varied to study the effect on drug transport. It was observed that the maximum concentration at the epithelial surface for the PE and the MM increased by 397 % and 380 %, respectively, when the peristaltic wave speed was decreased from 1.5 to 0.5 cm s−1. At this wave speed, physiologically relevant concentrations of PE were found at the epithelial surface. However, when the occlusion ratio is increased from 0.3 to 0.7, the concentration approaches zero. These results suggest that a slower-moving and more contracted peristaltic wave leads to higher efficiency in transporting mass to the epithelial wall during the peristalsis phases of the migrating motor complex.

Schizophrenia is a severe psychiatric disorder affecting approximately 20 million people worldwide. The disease consists of positive symptoms e.g. hallucinations, negative symptoms such as anhedonia, and cognitive deficits, e.g. impaired episodic memory. Most of the currently available treatment options for schizophrenia only target the positive symptoms, possess severe side effects and do not work for a large group of patients. In this thesis, the unique antipsychotic drug lumateperone and adjunctive treatment of sodium nitroprusside (SNP) to sub-maximal doses of conventional antipsychotic drugs are investigated in preclinical tests as novel treatment options for schizophrenia In paper I we showed that SNP enhances the antipsychotic-like effect of a sub-effective dose of risperidone in the conditioned avoidance response (CAR) test in rats. Moreover, by using microdialysis we showed that SNP significantly enhances risperidone-induced dopamine release in the rat medial prefrontal cortex (mPFC) but not in the nucleus accumbens, indicating that adjunct SNP could be used to improve the efficacy of antipsychotic drugs, while reducing their dose and subsequently lower the risk of side effects. In paper II we used microdialysis combined to the behavioral novel object recognition test in rats to show that the release of both dopamine and norepinephrine is increased in the ventral hippocampus in response to a novel object, suggesting that dopamine and norepinephrine may play a crucial role in recognition memory. In paper III we showed that SNP significantly enhanced the antipsychotic-like effect of sub-effective doses of olanzapine in the CAR test, but not of clozapine, this could be explained by the developed tolerance towards clozapine after repeated administrations. In paper IV we used enzyme-coated microelectrode arrays to show that lumateperone significantly increased cortical glutamate release in the mPFC of anaesthetized rats. By using electrophysiology, we also show that lumateperone facilitates NMDA and AMPA-mediated currents in a dopamine D1 dependent manner in layer V/VI pyramidal neurons in the mPFC of rats. Moreover, lumateperone increases dopamine release in the mPFC of freely moving rats as shown by using microdialysis. These mechanisms may improve cognitive deficits and contribute to the clinically demonstrated antidepressant effects of lumateperone. Taken together, these results show that lumateperone is a promising novel treatment option for schizophrenia, and that adjunct SNP treatment may allow for improved efficacy at maintained or even reduced dosage of conventional antipsychotic medication.

Solute carriers consist of over 400 genes and 66 subfamilies and are the largest group of membrane bound transporters. They facilitate movement of many molecules through cells and organelles and are crucial for upholding many key functions in the human body. They are also involved in the pathophysiology of several diseases. The sodium-coupled neutral amino acid transporter family (SLC38) consists of 11 known transporters classified into system A and system N. They are responsible for influx and efflux of amino acids in different tissues, and the broad majority are expressed in various brain regions. The transporters are involved in gamma-aminobutyric acid (GABA)/glutamine metabolism, liver metabolism, the mammalian target of rapamycin pathway, among many other pathways and mechanisms. This study quantitatively investigates the mRNA expression of the transporters in human nerve cells from differentiation day 0 to 30, with the main focus being on the Golgi/ER localized SLC38A10. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to run quantitative assays on nerve cells from day 0, 5, 10, 15 and 30 in nerve cell differentiation, with histone variant H3.3 and Ribosomal Protein L13a as housekeeping genes. It was found that the genes SLC38A1, A2, A3, A8 and A10 had relatively similar expression patterns in the nerve cell tissues tested. No expression was detected for SLC38A5, A6, A9 and A11. SLC38A4 had no clear expression pattern in the nerve cell tissues, although expression was detected in some assays, which was not expected in nerve cells. SLC38A10 demonstrated the highest expression, and lowest expression levels were seen for SLC38A4. SLC38A1, A2, A3, A8 and A10 had highest expression in day 0 nerve cells. These results indicate that precursor neurons could be most suitable to use for knockout studies of SLC38A10, as well as for SLC38A1, A2, A3 and A8, as these cells demonstrated the highest levels of expression of these genes. Further studies are encouraged into the expression levels of the SLC38 transporters, their roles in the human body and disease pathophysiology, and, ultimately, their potential as drug targets.

BACKGROUND: Hereditary angioedema (HAE) is caused by low levels of or defects in C1 inhibitor. Although disease activity may be modified by prophylaxis, emergency treatment, treatment for comorbidities, and oral contraceptives, the extent of their use is unclear. OBJECTIVE: To investigate trends in the use of disease-specific and interfering drugs in patients with HAE compared with the general population in Sweden. METHODS: In a nationwide, longitudinal study, 239 patients with HAE and 2 383 controls were compared with the Prescribed Drug Register (2005e2019). These data reflect rates of dispensed prescriptions from pharmacies in Sweden. RESULTS: Attenuated androgens were used by approximately 10% of patients with HAE. The number of individuals treated with prophylactic plasma-derived C1 inhibitor increased during this period to reach almost 25% in men and 35% in women in 2019. Tranexamic acid was prescribed to 5% to 15% of patients, primarily children and young adults. Rates of prescriptions for icatibant, an emergency medication, showed a steady increase since its introduction in 2010, in particular among middle-aged women, suggesting poorly controlled disease. The use of di-uretics, calcium channel blockers, and gestagens was more common in patients with HAE than in controls, whereas angiotensin-converting enzyme inhibitors were rarely collected. CONCLUSIONS: Despite concerns regarding side effects, approximately 10% of patients with HAE received attenuated androgens for long-term prophylaxis. The common use of emergency medication also suggests poorly controlled disease in many patients, highlighting the need for increased focus on prophylactic treatment.

Background Discrepancies in medication lists are common and can contribute to drug-related problems. This study was performed before the implementation of the National Medication List in Sweden, an intervention expected to improve the accuracy of medication lists. Aim The aim of the study was to examine the number and type of discrepancies in the medication list from pharmacies in Sweden. The secondary aim was to describe the information sources Swedish patients used as their medication lists and how confident they were with the information. Method Structured interviews were conducted with patients at 13 community pharmacies in Sweden during the period October 5, 2020, to April 16, 2021. The printed medication list was reviewed together with the patient to identify any discrepancies and missing information. Results A total of 327 patients were included in the study (response rate 51%). The printed medication list from pharmacies was the most common information source for patients to know which medications to use. Two thirds (n = 215) of the patients had at least one discrepancy among their prescriptions and 32% (n = 106) were missing at least one prescription medication. Among all prescriptions (n = 2567) 10% (n = 264) were non-current prescriptions, 9% (n = 238) were duplicates and 3% (n = 88) had the wrong dose. The proportion of prescriptions with discrepancies differed between drug-groups. Conclusion The discrepancies described in this study can have serious consequences, and results provide a baseline for studies after the implementation of the National Medication List.

Gambling disorder (GD) is a behavioral addiction characterized by persistent and recurrent gambling behavior that disrupts personal, social or professional life. Studies have revealed that GD shares many features with alcohol and substance use disorders, but little is known about potential unique features in GD and to what extent characteristics are shared. One shared feature is reward-related decision-making and individuals with GD display deficits in decision-making. The rat gambling task (rGT) has been developed to enable preclinical studies of reward-related decision-making and underlying neurobiological mechanisms. The aim of this thesis was to explore individual differences in decision-making strategies in the rGT and underlying behavioral phenotypes and neurobiology. Paper I: three groups with different decision-making strategies in the rGT were identified: the strategic, risky and safe group. The rGT strategies were shown to be stable over time, even after multiple interruptions and other behavioral testing. Rats with risky rGT strategies had higher voluntary alcohol intake but not elevated sexual behavior. Naltrexone treatment resulted in an overall lowered motivation in the rGT but had no effect on choice behavior. Paper II: individual differences in gambling strategies were found in the rGT and corresponding strategy groups were replicated from Paper I. Moreover, brain functional connectivity was assessed using resting-state functional Magnetic Resonance Imaging. Differences in rGT strategies were associated with connectivity in regions in or associated with brain reward networks. Paper III: levels of neurotransmitters and metabolites were explored using matrix-assisted laser desorption/ionization mass spectrometry imaging in selected brain regions. The strategy groups were revealed to differ in levels of neurotransmitters and metabolites in regions of importance for decision-making and reward. Paper IV: decision-making strategies in humans, using the Iowa gambling task, and in rats, using the rGT, were explored. Results showed that most humans and rats learned to favor the advantageous choices and showed similar variability in individual choice preferences during end performance. This thesis has provided new information about individual decision-making strategies in the rGT and associations with other reward-related behaviors as well as neurobiology. Characterization of the strategy groups indicates a shared underlying mechanism between rGT strategies and alcohol intake but not natural rewards. Neurobiological differences in regions important for reward processing were also revealed. Lastly, similar variability in individual choice preference was found in humans and rats and it is concluded that both clinical and preclinical research would benefit from more detailed analyses on individual variations in decision-making.

The gold standard for pharmacometrics modeling, along with its modeling format, is currently NONMEM. In order to use other software, there is often a manual step of converting a model from one format to another, which is both time-consuming and causes manual errors. This project aimed to solve this problem by creating a conversion- and validation tool from NONMEM to two formats: nlmixr2 and RxODE2. These are both, unlike NONMEM, freely available and integrated into R. This was done by integrating the two tools (conversion and validation) into the program Pharmpy, which can extract model information from NONMEM's model format. For conversion, the model was read into Pharmpy and then, part by part, converted to the respective model format. The associated validation compared the predictions of the respective programs to see if they differed significantly from each other. The project showed that this type of conversion is possible, but the programs showed a greater difference than expected. Part of this can be explained by a rounding of parameter values in Pharmpy, but further analysis also indicated fundamental differences in the determination of the predictions between the programs. Larger differences in predictions are for instance oftentimes equidistant from the actual observation, meaning the predictions are presumably calculated differently. While not disproving the converted model, smaller discrepancies between the programs would indicate a more confident validation. In summary, the developed tools are considered useful for models where Pharmpy supports parsing of the model. If not for complete conversion, then at least for partial conversion with manual correction, which is also an improvement over an entirely manual workflow.

Aims: Increasing concentrations of dietary sugar results in a linear accumulation of triglycerides in male Drosophila, while inducing a U-shaped obesity response in their offspring. Here, using a combination of proteomics and small RNA (sRNA) sequencing, we aimed at understanding the molecular underpinning in sperm for such plasticity.Results: Proteomic analysis of seminal vesicles revealed that increasing concentrations of dietary sugar resulted in a bell-shaped induction of proteins involved in metabolic/redox regulation. Using stains and in vivo redox reporter flies, this pattern could be explained by changes in sperm production of reactive oxygen species (ROS), more exactly mitochondria-derived H2O2. By quenching ROS with the antioxidant N-acetyl cysteine and performing sRNA-seq on sperm, we found that sperm miRNA is increased in response to ROS. Moreover, we found sperm mitosRNA to be increased in high-sugar diet conditions (independent of ROS). Reanalyzing our previously published data revealed a similar global upregulation of human sperm mitosRNA in response to a high-sugar diet, suggesting evolutionary conserved mechanisms.Innovation: This work highlights a fast response to dietary sugar in mitochondria-produced H2O2 in Drosophila sperm and identifies redox-sensitive miRNA downstream of this event.Conclusions: Our data support a model where changes in the sperm mitochondria in response to dietary sugar are the primary event, and changes in redox homoeostasis are secondary to mitochondrial ROS production. These data provide multiple candidates for paternal intergenerational metabolic responses as well as potential biomarkers for human male fertility. Funding Agencies|Swedish Research Council [2015-03141]; Ragnar Soderbergs foundation; Knut and Alice Wallenberg foundation [2015.0165]

Cyclotides are an intriguing class of structurally stable circular miniproteins of plant origin with numerous potential pharmaceutical and agricultural applications. To investigate the occurrence of cyclotides in Sri Lankan flora, 50 medicinal plants were screened, leading to the identification of a suite of new cyclotides from Geophila repens of the family Rubiaceae. Cycloviolacin O2-like (cyO2-like) gere 1 and the known cyclotide kalata B7 (kB7) were among the cyclotides characterized at the peptide and/or transcript level together with several putative enzymes, likely involved in cyclotide biosynthesis. Five of the most abundant cyclotides were isolated, sequenced, structurally characterized, and screened in antimicrobial and cytotoxicity assays. All gere cyclotides showed cytotoxicity (IC50 of 2.0-10.2 mu M), but only gere 1 inhibited standard microbial strains at a minimum inhibitory concentration of 4-16 mu M. As shown by immunohistochemistry, large quantities of the cyclotides were localized in the epidermis of the leaves and petioles of G. repens. Taken together with the cytotoxicity and membrane permeabilizing activities, this implicates gere cyclotides as potential plant defense molecules. The presence of cyO2-like gere 1 in a plant in the Rubiaceae supports the notion that phylogenetically distant plants may have coevolved to express similar cytotoxic cyclotides for a specific functional role, most likely involving host defense.

The pharmaceutical development process starts with patient populations and their unmet therapeutic needs. Traditional pharmaceutical manufacturing of solid oral dosage forms is based on the strategy of one-size-fits-all. This is problematic, especially for patient populations with high patient-to-patient variability, as in pediatrics. Historically, pharmaceutical development has focused on the adult population, neglecting the therapeutic needs unique to children. As a result, there is a lack of age-appropriate formulations—available in acceptable dosage forms and suitable dosage strengths—for safe and efficient drug therapies for children. To address this, additive manufacturing, more commonly known as three-dimensional (3D) printing, has emerged as a flexible manufacturing platform for production of dosage forms based on patient needs. Interest in 3D printing for pharmaceutical production has grown rapidly; however, to date the research has mainly focused on water-soluble drugs not in need of more advanced drug delivery systems to enable oral absorption. The overall aim of this thesis was therefore to develop lipid-based drug delivery strategies for poorly soluble drugs to be 3D printed into personalized solid oral dosage forms. In the first part, an observational study was performed at a pediatric oncology ward, together with analysis of the age-appropriateness of the oral medications. Administration through enteral feeding tubes was identified as the main reason for manipulation of the solid dosage forms. Furthermore, active pharmaceutical ingredients requiring age-appropriate, personalized dosage forms were identified. In the next part, emulsion gels from emulsified lipid-based formulations stabilized by surfactants (surfactant-stabilized emulsion gels) or solid particles (Pickering emulsion gel) were developed to incorporate a poorly water-soluble model compound. The rheological properties of the emulsion gels were investigated. The developed emulsion gels were successfully 3D printed into solid oral dosage forms by semi-solid extrusion (SSE). In the last part, central quality control attributes, including both the printable formulation and the 3D-printed tablets, were studied in the 3D-printing method developed for one of the emulsion gels. The SSE 3D-printed tablets complied with standardized uniformity tests for both mass and drug content and demonstrated high dose accuracy and short-term storage stability. To conclude, a method for SSE 3D printing of emulsion gels into lipid tablets was developed with promising potential for personalized dosing of lipophilic drugs in a clinical setting.

Background: In older age polypharmacy is common due to a higher rate of chronic diseases. Taking multiple medications can result in drug related problems resulting in higher health care costs. OptiMEDs is an information and communications technology (ICT) guided medication review program that was created to improve the quality of the prescription of medicines in elderly. A pilot study of OptiMEDs was performed in 3 nursing homes in Belgium during 2019-2020. Aim: To update the explicit criteria behind OptiMEDs and to make the tool more specific. Method: Through a literature search of explicit criteria of potentially inappropriate prescribing (PIM-lists), newer versions published between 1st of January 2018 – 31st of October 2022 were sought. Redundancy of alerts was reduced by removing all the double alerts of PIMs. To minimize the number of alerts issued by the tool, several approaches to exemptions from and exclusion to PIMs were tested. To evaluate the improvement, results from the previous and new rule base were compared using the same dataset from the OptiMEDs feasibility trial. Results:OnlyonePIM-list(Beer’scriteria)hadanewversion,resultingin45PIMsbeingadded to the database and 33 PIMs were removed. Removal of the duplicates resulted in a database with 594 PIMs. With the OptiMEDs data set; The amount of patients having a PIM decreased with 25.1 % when removing EU(7)-list as a source. The amount of patient having at least one candidate for deprescribing decreased with 10.4 % when only keeping the deprescribing.org and RTCs as a source. Conclusion: The update probably made OptiMEDs more user-friendly as the number of PIMs decreased and the number of medications candidated for deprescribing decreased. However, an important further update is still needed, which is to include the clinical-based PIMs.

The gastrointestinal mucus is a hydrogel lining the luminal side of the gastrointestinal epithelium. Mucus is vital for gut homeostasis because it protects the epithelium from the noxious external environment. However, from a drug delivery perspective, drugs have to permeate through the mucus to reach the epithelium; therefore, mucus might pose a barrier to drug absorption. Most of the information about mucus derives from fundamental studies performed on rodents. However, information from larger preclinical animal species is highly warranted for improving study designs and guiding better interpretation of data from preclinical assessments. Furthermore, improved understanding of the nature of the gastrointestinal mucus would enable the development of in vitro mucus models with increased biorelevance. These could then be implemented in drug absorption assays to improve the (bio)predictability. Well-informed in vitro mucus models would enable quick and less variable screening of drug candidates in the early drug development stages. Finally, these models would contribute to reduction, refinement, and replacement (the 3Rs) of animal usage in the drug development process. This thesis aims to improve our understanding of the nature of gastrointestinal mucus and its impact on drug absorption. For this purpose, mucus from the complete gastrointestinal tract of pigs and dogs was characterized and the diffusion of physicochemically diverse FITC-dextrans through colonic mucus was studied, both ex vivo and in vitro. The characterization of the gastrointestinal mucus focused on properties relevant for drug absorption and revealed the physiological characteristics, composition, and structural profiles from the various gastrointestinal regions. The findings pointed towards substantial differences between small intestinal and colonic mucus in both species and served as the basis for developing artificial colonic mucus models for drug permeation assessments. Porcine and canine artificial mucus models were developed and validated against the respective native secretions in terms of structural properties and demonstrated their potential to capture the key diffusion patterns of FITC-dextrans observed in native colonic mucus. Overall, this work provided insights into key properties of mucus from large preclinical species and validated tools for the assessment of the impact of mucus on drug absorption.

Introduction and Hypothesis: Patients with overactive bladder (OAB) often undergo prolonged treatment with one or more oral OAB medications. OnabotulinumtoxinA (onabotA), a type A botulinum toxin, may provide an appropriate alternative to oral treatments in patients intolerant of or refractory to one or more oral OAB medications. The GRACE study demonstrated real-world benefits of onabotA treatment for OAB in patients refractory to oral medications. This exploratory post hoc analysis of data from the GRACE study aims to determine if treatment history impacts benefit from treatment with onabotA. Methods: This is a subanalysis of the GRACE study, a prospective observational study (NCT02161159) that enrolled patients with symptomatic OAB inadequately managed by at least one oral OAB medication. Patients had a treatment history of one or more anticholinergics (AC) and/or β-3 adrenoreceptor agonists (β-3) for relief of OAB; results were stratified according to treatment history. Patients in this analysis elected to discontinue oral medications upon treatment with onabotA. Safety was followed for 12 months in all patients that received at least 1 dose of onabotA; efficacy was determined over a 12-week period. Results: Compared to baseline levels, significant reductions in urinary incontinence (UI), urgency, micturition, and nocturia were noted as early as 1 week and were sustained at 12 weeks, regardless of the type and number of oral medications taken before treatment with onabotA. At 12 weeks post-onabotA, the mean change from baseline UI episodes/day for those with a treatment history of only one AC was −2.4 (n = 43, p ≤ 0.001); more than one AC, −2.4 (n = 52, p ≤ 0.001); one β-3, −3.3 (n = 12, p < 0.05); at least one AC and at least one β-3, −3.2 (n = 56, p ≤ 0.001). Pad and liner use was significantly decreased at 12 weeks post-onabotA across all treatment history groups. Reductions in diaper pant use varied, with less of a reduction in patients with a treatment history of more than one AC compared to patients with a history of at least one AC and one β-3 (p < 0.05) or those with a history of only one AC (p < 0.05). Overall, a total of 253/288 of patients (88%) reported improvements on the treatment benefit scale 12 weeks after treatment with onabotA, regardless of type and number of prior oral medications. In the population of patients that received at least one dose of onabotA (N = 504), 57 adverse events were reported in 38 patients (7.5%); 9 were serious (1.8%). Urinary retention was reported in 5 patients (1.0%); 1 was severe (0.2%). Symptomatic urinary tract infection was reported in 2 patients (0.4%). Conclusions: In this exploratory post hoc analysis of real-world data from the GRACE study, there were few significant differences in outcomes based on the type and number of prior oral medications. Thus, patients who are refractory to one or more oral OAB medications may benefit from earlier treatment with onabotA.

Acetylbenzylfentanyl, benzoylbenzylfentanyl, 3-fluoro-methoxyacetylfentanyl, and 3-phenylpropanoylfentanyl are fentanyl analogs that have been reported to the European Monitoring Centre for Drugs and Drug Addiction in recent years. The aim of this study was to identify metabolic pathways and potential biomarker metabolites of these fentanyl analogs. The compounds were incubated (5 mu M) with cryopreserved hepatocytes for up to 5 h in vitro. Metabolites were analyzed with liquid chromatography-quadrupole time of flight-high-resolution mass spectrometry (LC-QTOF-HRMS). The experiments showed that acetylbenzylfentanyl, benzoylbenzylfentanyl, and 3-phenylpropanoylfentanyl were mainly metabolized through N-dealkylation (forming nor-metabolites) and 3-fluoro-methoxyacetylfentanyl mainly through demethylation. Other observed metabolites were formed by mono-/dihydroxylation, dihydrodiol formation, demethylation, dehydrogenation, amide hydrolysis, and/or glucuronidation. The experiments showed that a large number of metabolites of 3-phenylpropanoylfentanyl were formed. The exact position of hydroxy groups in formed monohydroxy metabolites could not be established solely based upon recorded MSMS spectra of hepatocyte samples. Therefore, potential monohydroxy metabolites of 3-phenylpropanoylfentanyl, with the hydroxy group in different positions, were synthesized and analyzed together with the hepatocyte samples. This approach could reveal that the beta position of the phenylpropanoyl moiety was highly favored; beta-OH-phenylpropanoylfentanyl was the most abundant metabolite after the nor-metabolite. Both metabolites have the potential to serve as biomarkers for 3-phenylpropanoylfentanyl. The nor-metabolites of acetylbenzylfentanyl, benzoylbenzylfentanyl, and 3-fluoro-methoxyacetylfentanyl do also seem to be suitable biomarker metabolites, as do the demethylated metabolite of 3-fluoro-methoxyacetylfentanyl. Identified metabolic pathways and formed metabolites were in agreement with findings in previous studies of similar fentanyl analogs. Funding Agencies|Eurostars-2 Joint Programme (European Commission) [E! 113377]; European Unions Horizon 2020 research and innovation program; Swedens Innovation Agency VINNOVA [2019-03566]; Strategic Research Area in Forensic Sciences (Strategiomradet forensiska vetenskaper) at Linkoeping University

The blood–brain barrier (BBB) poses major challenges to drug delivery to the CNS. SFTI-1 and kalata B1 are cyclic cell-penetrating peptides (cCPPs) with high potential to be used as scaffolds for drug delivery. We here studied their transport across the BBB and distribution within the brain to gauge the potential of these two cCPPs as scaffolds for CNS drugs. In a rat model, SFTI-1 exhibited, for a peptide, high extent of BBB transport with a partitioning of unbound SFTI-1 across the BBB, Kp,uu,brain, of 13%, while only 0.5% of kalata B1 equilibrated across the BBB. By contrast, kalata B1, but not SFTI-1, readily entered neural cells. SFTI-1, but not kalata B1, could be a potential CNS delivery scaffold for drugs directed to extracellular targets. These findings indicate that differences between the BBB transport and cellular uptake abilities of CPPs are crucial in the development of peptide scaffolds.

Globally, hepatocellular carcinoma (HCC) is the most common form of liver cancer and a leading cause of cancer death. One important risk factor is liver cirrhosis and the disease progression is characterized by deposition of extracellular matrix proteins that form a fibrous network, which increases liver stiffness and may limit the effectiveness of different treatment strategies. The overall aim of this thesis was to investigate the anticancer drug doxorubicin (DOX) and its clinically relevant drug delivery systems from an in vitro perspective. The focus was on developing and using qualitative and quantitative methods to better understand formulation performance, intracellular uptake and molecular diffusion. The experimental in vitro findings were then translated to clinical scenarios using physiologically based pharmacokinetic (PBPK) modelling. The performance of clinically employed emulsion formulations containing DOX and the tumor accumulating oil Lipiodol® were evaluated in terms of their stability (Paper I). The most stable emulsion (> 72 h) was achieved when using an aqueous phase containing the contrast agent iohexol and with an aqueous to lipid phase ratio of 1:4 to assure formation of a water-in-oil emulsion. This was followed by a cell-based study (Paper II) where nanoformulated DOX was compared to DOX in solution in terms of tumor cell toxicity, intracellular DOX uptake and intracellular formation of the main active metabolite doxorubicinol (DOXol). DOX in solution was more potent in all investigated cell lines, where the most sensitive cells (HepG2) displayed IC50 values that were approximately 100 times lower than the most resistant cell line (SNU449). This was explained by the rapid intracellular uptake in HepG2 cells which was also confirmed with a complimentary miniaturized chip technique in Paper IV. In papers III and IV the focus was on molecular diffusion across biomimetic hydrogels mimicking tissue properties of cirrhotic liver and early stage HCC. The diffusion of DOX was significantly reduced in biomimetic gels as compared with more commonly used agarose gels, however the presence of human liver tumor cells did not significantly influence diffusion. Simulations using a developed PBPK and spatio-temporal tissue concentration model suggested that a liver tumor resembling SNU449 cells would not reach therapeutic exposure levels in a clinical scenario while the diffusion of DOX required further reduction by the tumor extracellular matrix in order to generate tumor concentration-time curves consistent with in vivo observations. This thesis contributes to an increased understanding of using DOX and its drug delivery systems as a treatment option for HCC. The approach of translating in vitro experimental data to clinical scenarios using modelling will grow in relevance as methods become more complex and data more bio-relevant.

The clinical support system Electronic Expert Support (EES) is available at all pharmacies in Sweden to examine electronic prescriptions when dispensing to prevent drug-related problems (DRPs). DRPs are common, and result in patient suffering and substantial costs for society. The aim of this research was to study the use of EES for the pediatric population (ages 0–12 years), by describing what types of alerts are generated for potential DRPs, how they are handled, and how the use of EES has changed over time. Data on the number and categories of EES analyses, alerts, and resolved alerts were provided by the Swedish eHealth Agency. The study shows that the use of EES has increased. The most common type of alert for a potential DRP among pediatric patients was regarding high doses in children (30.3% of all alerts generated). The most common type of alert for a potential DRP that was resolved among pediatrics was therapy duplication (4.6% of the alerts were resolved). The most common reason for closing an alert was dialogue with patient for verification of the treatment (66.3% of all closed alerts). Knowledge of which type of alerts are the most common may contribute to increased prescriber awareness of important potential DRPs.

Contains fulltext : 290623.pdf (Publisher’s version ) (Open Access) Blood pressure measurements form a critical component of adverse event monitoring for tyrosine kinase inhibitors, but might also serve as a biomarker for dose titrations. This study explored the impact of various sources of within-individual variation on blood pressure readings to improve measurement practices and evaluated the utility for individual- and population-level dose selection. A pharmacokinetic-pharmacodynamic modeling framework was created to describe circadian blood pressure changes, inter- and intra-day variability, changes from dipper to non-dipper profiles, and the relationship between drug exposure and blood pressure changes over time. The framework was used to quantitatively evaluate the influence of physiological and pharmacological aspects on blood pressure measurements, as well as to compare measurement techniques, including office-based, home-based, and ambulatory 24-h blood pressure readings. Circadian changes, as well as random intra-day and inter-day variability, were found to be the largest sources of within-individual variation in blood pressure. Office-based and ambulatory 24-h measurements gave rise to potential bias (>5 mmHg), which was mitigated by model-based estimations. Our findings suggest that 5-8 consecutive, home-based, measurements taken at a consistent time around noon, or alternatively within a limited time frame (e.g., 8.00 a.m. to 12.00 p.m. or 12.00 p.m. to 5.00 p.m.), will give rise to the most consistent blood pressure estimates. Blood pressure measurements likely do not represent a sufficiently accurate method for individual-level dose selection, but may be valuable for population-level dose identification. A user-friendly tool has been made available to allow for interactive blood pressure simulations and estimations for the investigated scenarios. 01 januari 2023

When drugs need to be delivered to the respiratory system dry powder inhalers (DPIs) are often used. The powders in DPIs often consists of a micronized active pharmaceutical ingredient (API) that is adhered to a larger carrier particle forming an adhesive mixture. To achieve a good drug delivery aerosolization is very important and to improve the aerosolization a force control agent such as magnesium stearate (MgSt) may be used. The aim of this project is to investigate how MgSt effects the aerosolization and bulk properties of adhesive mixtures containing different amount of API. Several analyses of bulk properties such as density, bulk density and compressibility were made. The results from these experiments showed that the only bulk properties that had a significant change when MgSt was added was the bulk density. When investigating if the aerosolization properties changed with added MgSt a fast-screening impactor (FSI) and a next generation impactor (NGI) were used. The results showed a clear improvement of aerosolization properties for all the powder containing MgSt. The fine particle fraction (FPF) and fine particle dose (FPD) both increased for all the powder when MgSt was added. The pressure drop sensitivity decreased when MgSt was added which also was desired. In conclusion adding MgSt seems to be beneficial for the aerosolization properties regardless of the amount of API in the powder.

This study aimed to evaluate a material sparing method to predict tabletability and compactibility relationships. Seven a-lactose monohydrate powders with varying particle size were used as test materials. The compressibility of the powders was determined experimentally, whereas tabletability and compactibility profiles were derived both experimentally and predicted. In the prediction method, two experimental compression parameters (Kawakita b-1 and Heckel plastic stiffness) and a single tensile strength reference value were used, all necessary data obtained from a single compression experiment. For both predicted and experimental relationships, compaction and tableting parameters (performance indicators) were calculated. The correction for viscoelastic recovery was successful in generating compressibility profiles that corresponded to the series of experimental out-of-die tablet porosities. For both the tabletability and compactibility, the experimental and predicted profiles showed a high degree of similarity. Good correlations were obtained between the predicted and experimental compaction and tableting parameters. It is concluded that the hybrid prediction method is a material sparing method, which can give good approximations of tabletability and compactibility relationships. The prediction method has the potential to be included as a part of a protocol for the characterisation of the tableting performance of particulate solids.

The emergence of multidrug-resistant (MDR) pathogens and the gradual depletion of available antibiotics have exacerbated the need for novel antimicrobial agents with minimal toxicity. Herein, we report functionally substituted pyridine carbohydrazide with remarkable antimicrobial effect on multi-drug resistant strains. In the series, compound 6 had potent activity against four MDR strains of Candida spp., with minimum inhibitory concentration (MIC) values being in the range of 16–24 µg/mL and percentage inhibition up to 92.57%, which was exceptional when compared to broad-spectrum antifungal drug fluconazole (MIC = 20 µg/mL, 81.88% inhibition). Substitution of the octyl chain in 6 with a shorter butyl chain resulted in a significant anti-bacterial effect of 4 against Pseudomonas aeruginosa (ATCC 27853), the MIC value being 2-fold superior to the standard combination of ampicillin/cloxacillin. Time-kill kinetics assays were used to discern the efficacy and pharmacodynamics of the potent compounds. Further, hemolysis tests confirmed that both compounds had better safety profiles than the standard drugs. Besides, molecular docking simulations were used to further explore their mode of interaction with target proteins. Overall results suggest that these compounds have the potential to become promising antimicrobial drugs against MDR strains.

Tenofovir alafenamide (TAF) is an antiretroviral (ARV) drug that is used for the management and prevention of human immunodeficiency virus (HIV). The clinical availability of ARV delivery systems that provide long-lasting protection against HIV transmission is lacking. There is a dire need to formulate nanocarrier systems that can help in revolutionizing the way to fight against HIV/AIDS. Here, we aimed to synthesize a polymer using chitosan and polyethylene glycol (PEG) by the PEGylation of chitosan at the hydroxyl group. After successful modification and confirmation by FTIR, XRD, and SEM, TAF-loaded PEGylated chitosan nanoparticles were prepared and analyzed for their particle size, zeta potential, morphology, crystallinity, chemical interactions, entrapment efficacy, drug loading, in vitro drug release, and release kinetic modeling. The fabricated nanoparticles were found to be in a nanosized range (219.6 nm), with similar to 90% entrapment efficacy, similar to 14% drug loading, and a spherical uniform distribution. The FTIR analysis confirmed the successful synthesis of PEGylated chitosan and nanoparticles. The in vitro analysis showed similar to 60% of the drug was released from the PEGylated polymeric reservoir system within 48 h at pH 7.4. The drug release kinetics were depicted by the Korsmeyer-Peppas release model with thermodynamically nonspontaneous drug release. Conclusively, PEGylated chitosan has the potential to deliver TAF from a nanocarrier system, and in the future, cytotoxicity and in vivo studies can be performed to further authenticate the synthesized polymer.

Background Apramycin is under development for human use as EBL-1003, a crystalline free base of apramycin, in face of increasing incidence of multidrug-resistant bacteria. Both toxicity and cross-resistance, commonly seen for other aminoglycosides, appear relatively low owing to its distinct chemical structure. Objectives To perform a population pharmacokinetic (PPK) analysis and predict an efficacious dose based on data from a first-in-human Phase I trial. Methods The drug was administered intravenously over 30 min in five ascending-dose groups ranging from 0.3 to 30 mg/kg. Plasma and urine samples were collected from 30 healthy volunteers. PPK model development was performed stepwise and the final model was used for PTA analysis. Results A mammillary four-compartment PPK model, with linear elimination and a renal fractional excretion of 90%, described the data. Apramycin clearance was proportional to the absolute estimated glomerular filtration rate (eGFR). All fixed effect parameters were allometrically scaled to total body weight (TBW). Clearance and steady-state volume of distribution were estimated to 5.5 L/h and 16 L, respectively, for a typical individual with absolute eGFR of 124 mL/min and TBW of 70 kg. PTA analyses demonstrated that the anticipated efficacious dose (30 mg/kg daily, 30 min intravenous infusion) reaches a probability of 96.4% for a free AUC/MIC target of 40, given an MIC of 8 mg/L, in a virtual Phase II patient population with an absolute eGFR extrapolated to 80 mL/min. Conclusions The results support further Phase II clinical trials with apramycin at an anticipated efficacious dose of 30 mg/kg once daily.

Alpinia galanga Willd., greater galangal, has been used for thousands of years as a spice as well as in traditional medicine. Its central nervous system (CNS) stimulant activity and neuroprotective effects have been proved both in animal models and human trials. However, the compounds responsible for these effects have not been identified yet. Therefore, the main constituents (p-OH-benzaldehyde (1), trans-p-coumaryl-alcohol (2), p-coumaryl-aldehyde (4), galanganol A (5), galanganol B (6), trans-p-acetoxycinnamyl alcohol (7), 1′S-1′-acetoxychavicol acetate (ACA, 9), and 1′S-1′-acetoxyeugenol acetate (AEA, 10)) were isolated to investigate their aqueous stability and passive diffusion across the gastro-intestinal tract (GIT) membrane and the blood–brain barrier (BBB) by the parallel artificial membrane permeability assay (PAMPA). Our positive results for compounds 1, 2, 4, 7, 9, and 10 suggest good permeability, thus potential contribution to the effects of greater galangal in the CNS. The results of the PAMPA-BBB were corroborated by in silico chemography-based ChemGPS-NP framework experiments. In addition, examination of the chemical space position of galangal compounds in relation to known psychostimulants revealed that all the molecules in proximity are NET/SERT inhibitors. As ACA and AEA did not show much proximity to either compound, the importance of further investigation of their degradation products becomes more pronounced.

Transcellular permeation enhancers are known to increase the intestinal permeability of enalaprilat, a 349 Da peptide, but not hexarelin (887 Da). The primary aim of this paper was to investigate if paracellular permeability enhancers affected the intestinal permeation of the two peptides. This was investigated using the rat single-pass intestinal perfusion model with concomitant blood sampling. These luminal compositions included two paracellular permeation enhancers, chitosan (5 mg/mL) and ethylenediaminetetraacetate (EDTA, 1 and 5 mg/mL), as well as low luminal tonicity (100 mOsm) with or without lidocaine. Effects were evaluated by the change in lumen-to-blood permeability of hexarelin and enalaprilat, and the blood-to-lumen clearance of 51chromium-labeled EDTA (CLCr-EDTA), a clinical marker for mucosal barrier integrity. The two paracellular permeation enhancers increased the mucosal permeability of both peptide drugs to a similar extent. The data in this study suggests that the potential for paracellular permeability enhancers to increase intestinal absorption of hydrophilic peptides with low molecular mass is greater than for those with transcellular mechanism-of-action. Further, the mucosal blood-to-lumen flux of 51Cr-EDTA was increased by the two paracellular permeation enhancers and by luminal hypotonicity. In contrast, luminal hypotonicity did not affect the lumen-to-blood transport of enalaprilat and hexarelin. This suggests that hypotonicity affects paracellular solute transport primarily in the mucosal crypt region, as this area is protected from luminal contents by a constant water flow from the crypts., Graphical abstract Paracellular permeation enhancers increased the intestinal transport of the peptide drug hexarelin but there was no effect of transcellular permeation enhancers.Image 1

Drug absorption following oral administration is determined by complex and dynamic interactions between gastrointestinal (GI) physiology, the drug, and its formulation. Since many of these interactions are not fully understood, the COST action on "Understanding Gastrointestinal Absorption-related Processes (UNGAP)" was initiated in 2017, with the aim to improve the current comprehension of intestinal drug absorption and foster future developments in this field. In this regard, in vivo techniques used for the characterization of human GI physiology and the intraluminal behavior of orally administered dosage forms in the GI tract are fundamental to gaining deeper mechanistic understanding of the interplay between human GI physiology and drug product performance. In this review, the potential applications, advantages, and limitations of the most important in vivo techniques relevant to oral biopharmaceutics are presented from the perspectives of different research fields. ispartof: PHARMACEUTICS vol:14 issue:4 ispartof: location:Switzerland status: published

Apramycin represents a subclass of aminoglycoside antibiotics that has been shown to evade almost all mechanisms of clinically relevant aminoglycoside resistance. Model-informed drug development may facilitate its transition from preclinical to clinical phase. This study explored the potential of pharmacokinetic/pharmacodynamic (PK/PD) modeling to maximize the use of in vitro time-kill and in vivo preclinical data for prediction of a human efficacious dose (HED) for apramycin. PK model parameters of apramycin from four different species (mouse, rat, guinea pig, and dog) were allometrically scaled to humans. A semimechanistic PK/PD model was developed from the rich in vitro data on four Escherichia coli strains and subsequently the sparse in vivo efficacy data on the same strains were integrated. An efficacious human dose was predicted from the PK/PD model and compared with the classical PK/PD index methodology and the aminoglycoside dose similarity. One-compartment models described the PK data and human values for clearance and volume of distribution were predicted to 7.07 L/hour and 26.8 L, respectively. The required fAUC/MIC (area under the unbound drug concentration-time curve over MIC ratio) targets for stasis and 1-log kill in the thigh model were 34.5 and 76.2, respectively. The developed PK/PD model predicted the efficacy data well with strain-specific differences in susceptibility, maximum bacterial load, and resistance development. All three dose prediction approaches supported an apramycin daily dose of 30 mg/kg for a typical adult patient. The results indicate that the mechanistic PK/PD modeling approach can be suitable for HED prediction and serves to efficiently integrate all available efficacy data with potential to improve predictive capacity.

This article highlights some numerical challenges when implementing the bounded integer model for composite score modeling and suggests an improved implementation. The improvement is based on an approximation of the logarithm of the error function. After presenting the derivation of the improved implementation, the article compares the performance of the algorithm to a naive implementation of the log-likelihood using both simulations and a real data example. In the simulation setting, the improved algorithm yielded more precise and less biased parameter estimates when the within-subject variability was small and estimation was performed using the Laplace algorithm. The estimation results did not differ between implementations when the SAEM algorithm was used. For the real data example, bootstrap results differed between implementations with the improved implementation producing identical or better objective function values. Based on the findings in this article, the improved implementation is suggested as the new default log-likelihood implementation for the bounded integer model. Electronic supplementary material The online version of this article (10.1007/s10928-020-09727-8) contains supplementary material, which is available to authorized users.

Background Buprenorphine is one of the most used analgesics for postoperative pain in rabbits. The recommended dose in rabbits (0.01–0.05 mg/kg) is the same for intravenous (IV), intramuscular (IM), and subcutaneous (SC) administration, despite lack of pharmacokinetic data. Five male and five female New Zealand White rabbits (mean ± SD body weight 3.1 ± 0.3 kg) were administered 0.05 mg/kg buprenorphine by the IV, IM and SC routes and 0.1 mg/kg by the SC route, in a cross-over design with two-week wash-out periods between treatments. Blood was collected before, and up to 8 h post buprenorphine injection, for determination of serum levels by UPHLC-MS/MS. Results The area under the time concentration curve (AUC0-t) was lower after SC (398 ± 155 ng/mL/min) than IM (696 ± 168 ng/mL/min, p p p = 0.006). The elimination half-life was longer after SC (260 ± 120 min) than after IM (148 ± 26 min, p = 0.002) as well as IV (139 ± 33 min) injection (p p = 0.022) and 165% in male rabbits (p p = 0.6), whereas it increased in the males (71 ± 23%, p = 0.008). Conclusions The lower bioavailability of 0.05 mg/kg buprenorphine after SC administration could explain the lack of efficacy seen in clinical pain studies in rabbits, using this route. For immediate pain relief, IV or IM administration is therefore be recommended, whereas SC administration may be useful to sustain analgesic serum levels, once efficient pain relief has been achieved. The current data do not support an increase in dose to compensate for the lower SC bioavailability.

Polyphenols are believed to possess numerous health benefits and can be grouped as phenolic acids, flavonoids or non-flavonoids. Research involving the synthesis of nanopolyphenols has attracted interest in the areas of functional food, nutraceutical and pharmaceutical development. This is in an effort to overcome current challenges which limit the application of polyphenols such as their rapid elimination, low water-solubility, instability at low pH, and their particle size. In the synthesis of nanopolyphenols, the type of nanocarrier used, the nanoencapsulation technique employed and the type of polymers that constitute the drug delivery system are crucial. For this review, all mentioned factors which can influence the therapeutic efficacy of nanopolyphenols were assessed. Their efficacy as anti-diabetic agents was also evaluated in 33 publications. Among these were phenolic acid (1), flavonoids (13), non-flavonoids (17) and polyphenol-rich extracts (2). The most researched polyphenols were quercetin and curcumin. Nanoparticles were the main nanocarrier and the size of the nanopolyphenols ranged from 15 to 333 nm with encapsulation efficiency and drug loading capacities of 56–97.7% and 4.2–53.2%, respectively. The quantity of nanomaterial administered orally ranged from 1 to 300 mg/kg/day with study durations of 1–70 days. Most studies compared the effect of the nanopolyphenol to its free-form and, in all but three cases, significantly greater effects of the former were reported. Assessment of the polyphenol to understand its properties and the subsequent synthesis of its nanoencapsulated form using suitable nanocarriers, polymers and encapsulation techniques can result in effective therapeutic agents for the treatment of diabetes.

Purpose To describe 5-fluorouracil (5FU) pharmacokinetics, myelotoxicity and respective covariates using a simultaneous nonlinear mixed effect modelling approach. Methods Thirty patients with gastrointestinal cancer received 5FU 650 or 1000 mg/m2/day as 5-day continuous venous infusion (14 of whom also received cisplatin 20 mg/m2/day). 5FU and 5-fluoro-5,6-dihydrouracil (5FUH2) plasma concentrations were described by a pharmacokinetic model using NONMEM. Absolute leukocyte counts were described by a semi-mechanistic myelosuppression model. Covariate relationships were evaluated to explain the possible sources of variability in 5FU pharmacokinetics and pharmacodynamics. Results Total clearance of 5FU correlated with body surface area (BSA). Population estimate for total clearance was 249 L/h. Clearances of 5FU and 5FUH2 fractionally changed by 77%/m2 difference from the median BSA. 5FU central and peripheral volumes of distribution were 5.56 L and 28.5 L, respectively. Estimated 5FUH2 clearance and volume of distribution were 121 L/h and 96.7 L, respectively. Baseline leukocyte count of 6.86 × 109/L, as well as mean leukocyte transit time of 281 h accounting for time delay between proliferating and circulating cells, was estimated. The relationship between 5FU plasma concentrations and absolute leukocyte count was found to be linear. A higher degree of myelosuppression was attributed to combination therapy (slope = 2.82 L/mg) with cisplatin as compared to 5FU monotherapy (slope = 1.17 L/mg). Conclusions BSA should be taken into account for predicting 5FU exposure. Myelosuppression was influenced by 5FU exposure and concomitant administration of cisplatin.

Aims To externally validate an earlier characterized relationship between bedaquiline exposure and decline in bacterial load in a more difficult-to-treat patient population, and to explore the performances of alternative dosing regimens through simulations. Methods The bedaquiline exposure-response relationship was validated using time-to-positivity data from 233 newly diagnosed or treatment-experienced patients with drug-resistant tuberculosis from the C209 open-label study. The significance of the exposure-response relationship on the bacterial clearance was compared to a constant drug effect model. Tuberculosis resistance type and the presence and duration of antituberculosis pre-treatment were evaluated as additional covariates. Alternative dosing regimens were simulated for tuberculosis patients with different types of drug resistance. Results High bedaquiline concentrations were confirmed to be associated with faster bacterial load decline in patients, given that the exposure-effect relationship provided a significantly better fit than the constant drug effect (relative likelihood = 0.0003). The half-life of bacterial clearance was identified to be 22% longer in patients with pre-extensively drug-resistant (pre-XDR) tuberculosis (TB) and 86% longer in patients with extensively drug-resistant (XDR) TB, compared to patients with multidrug-resistant (MDR) TB. Achievement of the same treatment response for (pre-)XDR TB patients as for MDR TB patients would be possible by adjusting the dose and dosing frequency. Furthermore, daily bedaquiline administration as in the ZeNix regimen, was predicted to be as effective as the approved regimen. Conclusion The confirmed bedaquiline exposure-response relationship offers the possibility to predict efficacy under alternative dosing regimens, and provides a useful tool for potential treatment optimization.

This review addresses questions on how to accomplish successful central nervous system (CNS) drug delivery (i.e., having the right concentration at the right CNS site, at the right time), by understanding the rate and extent of blood-brain barrier (BBB) transport and intra-CNS distribution in relation to CNS target site(s) exposure. To this end, we need to obtain and integrate quantitative and connected data on BBB using the Combinatory Mapping Approach that includes in vivo and ex vivo animal measurements, and the physiologically based comprehensive LEICNSPK3.0 mathematical model that can translate from animals to humans. For small molecules, slow diffusional BBB transport and active influx and efflux BBB transport determine the differences between plasma and CNS pharmacokinetics. Obviously, active efflux is important for limiting CNS drug delivery. Furthermore, liposomal formulations of small molecules may to a certain extent circumvent active influx and efflux at the BBB. Interestingly, for CNS pathologies, despite all reported disease associated BBB and CNS functional changes in animals and humans, integrative studies typically show a lack of changes on CNS drug delivery for the small molecules. In contrast, the understanding of the complex vesicle-based BBB transport modes that are important for CNS delivery of large molecules is in progress, and their BBB transport seems to be significantly affected by CNS diseases. In conclusion, today, CNS drug delivery of small drugs can be well assessed and understood by integrative approaches, although there is still quite a long way to go to understand CNS drug delivery of large molecules.

Background: This systematic review summarizes the impact of pharmacogenetics on the effect and safety of non-steroidal anti-inflammatory drugs (NSAIDs) and antidepressants when used for pain treatment. Methods: A systematic literature search was performed according to the preferred reporting items for systematic reviews and meta-analysis (PRISMA) guidelines regarding the human in vivo efficacy and safety of NSAIDs and antidepressants in pain treatment that take pharmacogenetic parameters into consideration. Studies were collected from PubMed, Scopus, and Web of Science up to the cutoff date 18 October 2021. Results: Twenty-five articles out of the 6547 initially detected publications were identified. Relevant medication-gene interactions were noted for drug safety. Interactions important for pain management were detected for (1) ibuprofen/CYP2C9; (2) celecoxib/CYP2C9; (3) piroxicam/CYP2C8, CYP2C9; (4) diclofenac/CYP2C9, UGT2B7, CYP2C8, ABCC2; (5) meloxicam/CYP2C9; (6) aspirin/CYP2C9, SLCO1B1, and CHST2; (7) amitriptyline/CYP2D6 and CYP2C19; (8) imipramine/CYP2C19; (9) nortriptyline/CYP2C19, CYP2D6, ABCB1; and (10) escitalopram/HTR2C, CYP2C19, and CYP1A2. Conclusions: Overall, a lack of well powered human in vivo studies assessing the pharmacogenetics in pain patients treated with NSAIDs or antidepressants is noted. Studies indicate a higher risk for partly severe side effects for the CYP2C9 poor metabolizers and NSAIDs. Further in vivo studies are needed to consolidate the relevant polymorphisms in NSAID safety as well as in the efficacy of NSAIDs and antidepressants in pain management.

Background: Drug utilization studies are essential to facilitate rational drug use in the society. Aim: In this review, we provide an overview of drug utilization measures that can be used with individual-level drug dispensing data, referencing additional reading on the individual analysis. This is intended to serve as a primer for those new to drug utilization research and a shortlist from which researchers can identify useful analytical approaches when designing their drug utilization study. Results and Discussion: We provide an overview of: (1) basic measures of drug utilization which are used to describe changes in drug use over time or compare drug use in different populations; (2) treatment adherence measures with specific focus on persistence and implementation; (3) how to measure drug combinations which is useful when assessing drug–drug interactions, concomitant treatment, and polypharmacy; (4) prescribing quality indicators and measures to assess variations in drug use which are useful tools to assess appropriate use of drugs; (5) proxies of prescription drug misuse and skewness in drug use; and (6) considerations when describing the characteristics of drug users or prescribers.

Positron emission tomography (PET), a medical imaging technique allowing for studies of the living human brain, has gained an important role in clinical trials of novel drugs against Alzheimer’s disease (AD). For example, PET data contributed to the conditional approval in 2021 of aducanumab, an antibody directed towards amyloid-beta (Aβ) aggregates, by showing a dose-dependent reduction in brain amyloid after treatment. In parallel to clinical studies, preclinical studies in animal models of Aβ pathology may also benefit from PET as a tool to detect target engagement and treatment effects of anti-Aβ drug candidates. PET is associated with a high level of translatability between species as similar, non-invasive protocols allow for longitudinal rather than cross-sectional studies and can be used both in a preclinical and clinical setting. This review focuses on the use of preclinical PET imaging in genetically modified animals that express human Aβ, and its present and potential future role in the development of drugs aimed at reducing brain Aβ levels as a therapeutic strategy to halt disease progression in AD. Title in Web of Science: PET Imaging in Preclinical Anti-A beta Drug Development

Oral administration is the preferred route for drug delivery because it has high patient compliance and is cost effective. The aqueous solubility of modern drug candidates is often poor, but a drug delivered orally must dissolve in the intestine so that it can be absorbed into the circulation. Advanced formulations can be used to improve the solubility, but the possible improvement in bioavailability from formulation varies among drug molecules. In vitro methods can be used to assess solubility, but these are slow and consume valuable drug material that is often rare early in development. As a non-destructive alternative, in silico methods have the potential to predict solubility, but methods available are in need of improvement, especially for predictions of drug solubility in intestinal fluid and for refining drug formulations. The goal of this thesis is to use molecular dynamics (MD) simulations to investigate the colloidal structures in intestinal fluids that affect drug solubility, and to simulate processes that affect solubility on the molecular level. Coarse-grained MD simulation protocols for biorelevant media, human and dog duodenal fluids, and lipid-based formulations were established based on concentrations measured in vivo. In the simulations, colloids self-assembled to micelles and vesicles depending on concentration and component input. Simulations with biorelevant media resulted in micelles qualitatively similar to those experimentally measured by small-angle X-rays. The structure of the colloids in the simulations were described in detail, and used to qualitatively assess drug solubility enhancement in model compounds with poor water solubility. These assessments were made by looking at the displacement of drugs and the drugs’ interactions with molecules in small intestinal fluid. The MD simulations were not able replace current solubility-predicting in silico models, but do show that coarse-grained MD simulations can be used for investigating the relevant processes involving intestinal fluids and lipid-based formulations.

Bedaquiline is a novel adenosine triphosphate synthase inhibitor anti-tuberculosis drug. Bedaquiline belongs to the class of diarylquinolines, which are antituberculosis drugs that are quite different mechanistically from quinolines and flouroquinolines. The fact that relatively similar chemical drugs produce different mechanisms of action is still not widely understood. To enhance discrimination in favor of bedaquiline, a new approach using eight-score principal component analysis (PCA), provided by a ChemGPS-NP model, is proposed. PCA scores were calculated based on 35 + 1 different physicochemical properties and demonstrated clear differences when compared with other quinolines. The ChemGPS-NP model provided an exceptional 100 compounds nearest to bedaquiline from antituberculosis screening sets (with a cumulative Euclidian distance of 196.83), compared with the different 2Dsimilarity provided by Tanimoto methods (extended connective fingerprints and the Molecular ACCess System, showing 30% and 182% increases in cumulative Euclidian distance, respectively). Potentially similar compounds from publicly available antituberculosis compounds and Maybridge sets, based on bedaquiline’s eight-dimensional similarity and different filtrations, were identified too.

Background: This thesis explores the pharmaceutical model physiologically based pharmacokinetics (PBPK) further and focuses on the basics of parameters used for the software tools to develop new drugs on the market today. Further reflects on what drug companies place their importance on when developing a new drug and why, by focusing on 29 different drugs and the parameters provided by the PBPK-model. Aim: To examine and compare 29 drugs, by focusing on comparing and evaluating administration route, formulation, elimination route, including metabolizing enzymes and parameters. These 4 focus points are a way to examine and reflect on what drug companies focus on when producing a new drug and their importance as to why they focus on what they do. Methods: Information on 29 drugs was collected from the PBPK model library in the Open system pharmacology file provided by the pharmacokinetic department of Uppsala University. Answering 9 points and then sorting and organizing the collected information with google sheets, making graphs and charts to present the result. Results: The most common administration route when creating PBPK-models was oral administration, the second most common was both oral and IV administration. The most common formulation was tablet, second most common was IV. The most common elimination route was trough the liver, with the help of enzymes. The most common one being enzyme CYP3A4, second was CYP2C19, and then CYP2C9. The most common and recurring parameters in order were lipophilicity, specific intestinal permeability, dissolution shape, specific intestinal permeability (trancellural), dissolution time (50% dissolved) and solubility at referenced pH. Conclusions: The result showed that the 4 focus points in the aim had a common thread and had reasonable results. They accurately reflected the drug companies' reasoning for producing and designing the drugs the way they did, such as being convenient and easy to take, adapted to the body’s physiology and being safe. The result showed the parameters as the most interesting section and formulation route a bit weaker and contributing to uncertain results for its section. Fantastisk presentation.

Nonlinear Mixed effect models are often used to describe population pharmacokinetics (PK) and Pharmacodynamics (PD) and play an important part of drug development both from regulatory and industry point of view. However, they can be time consuming and computationally expensive to develop. This thesis is a part of a larger collaboration between Uppsala University and two pharmaceutical companies, with the aim to develop a suite of software that can automate the model building process with more efficiency. One aspect that is important during the model building process is to detect how much the population parameter estimates are influenced by particular individuals. The results of this might lead to reconsideration of the model structure, as well as exclusion of these individuals from the dataset. The current tools available to detect this use case deletion diagnostics (CDD) to run the model multiple times with each subject removed from the dataset to examine whether the population estimates alter when that individual is removed. Another important aspect is whether an individual is an outlier from the population parameter predictions, which is obtained from simulating the model and evaluating the residuals (simeval). Both of these tools are computationally expensive and can take a lot of time, in particular CDD. Therefore, we developed a tool using machine learning (ML) algorithms that can predict these individuals based on other criteria, which will decrease the runtime in an automated model building procedure, whilst maintaining the robustness of the current methods described above. To create a training database for the ML models, predictors were extracted from 27 previously published models and the CDD and simeval diagnostic tools were run on these models to obtain that true values we want the ML model to predict. The database was then used to train two artificial neural networks (ANN) which is an efficient and powerful method in ML. To enable ‘on-the-fly’ predictions, the developed ANN models were deployed using tflite into pharmpy. The resulted ANNs were able to predict outlying individuals with 79% sensitivity, 83% precision, and 99.1% specificity. While the influential individuals ANN was able to predict with 58% sensitivity, 63% precision and 99.6% specificity. Both ANNs offered a rapid assessment of influential individuals and outlying individuals and were able to make predictions in a matter of sub-seconds compared to hours using traditional methods.