Browsing by Subject "drug development"

Sort by: Order: Results:

Now showing items 1-13 of 13
  • Karppinen, Jutta (Helsingfors universitet, 2017)
    In vitro liver cell models are important systems to study for example hepatotoxicity, which is one of the major causes for safety-related failures of drug candidates. 2D cell culture-based tests for compound screening are standard procedures in drug discovery, but reliable data for in vivo studies is hard to obtain because cells in a monolayer are in unnatural microenvironment. In turn, cells in 3D culture systems have more natural interactions with other cells and extracellular matrix, and their responses to drugs resemble more in vivo responses. In drug discovery and development, automation of the cell culture processes and compound screening saves time and costs, and improves the consistency and sterility of the procedures. As 3D cell culture systems are becoming more compatible with automation, they are also more promising to be used in drug discovery and development. The aim of the study was to develop and optimize automated processes for preparing 3D cell cultures into 96-well plates. HepG2, a human liver cancer cell line, cultures in nanofibrillar cellulose were prepared into well plates manually or by using automated liquid handling system. To our knowledge, this was the first time that automated processes for cell seeding into NFC were used to prepare 3D cell cultures. Cell seeding steps that could be automated were identified and optimized based on visual analysis of the wells and viability of the cells after seeding. After optimization, manual and automated processes were compared by studying cell viability, morphology and functionality. Alamar blue assay, Live/Dead assay and fluorescence-activated cell sorting were used to study cell viability, and F-actin staining, differential interference contrast microscopy and light microscopy were used to investigate cell morphology. Cell functionality was analyzed by studying albumin secretion. Cells seeded by using automation secreted normal amounts of liver-specific albumin. Cells maintained viability, morphology and functionality for four days after seeding although the results of viability varied. Alamar blue assays showed decreased development of viability although viability of manually seeded cells increased, but in other experiments the results from cultures seeded manually or by using automation were more similar. For example, lower viscosity of nanofibrillar cellulose and longer waiting time of cells at room temperature before automated processes are possible explanations for differences, as well as the natural variability in cell studies. In the future, automated high-throughput screening of compounds could be performed in 3D cell cultures prepared by using automation. That would save time and costs, and increase the correlation between in vitro and in vivo studies.
  • Jantti, Maria H.; Talman, Virpi; Räsänen, Kati; Tarvainen, Ilari; Koistinen, Hannu; Tuominen, Raimo K. (2018)
    Prostate cancer is one of the most common cancers in men. Although it has a relatively high 5-year survival rate, development of resistance to standard androgen-deprivation therapy is a significant clinical problem. Therefore, novel therapeutic strategies are urgently needed. The protein kinase C (PKC) family is a putative prostate cancer drug target, but so far no PKC-targeting drugs are available for clinical use. By contrast to the standard approach of developing PKC inhibitors, we have developed isophthalate derivatives as PKC agonists. In this study, we have characterized the effects of the most potent isophthalate, 5-(hydroxymethyl) isophthalate 1a3 (HMI-1a3), on three prostate cancer cell lines (LNCaP, DU145, and PC3) using both 2D and 3D cell culture models. In 2D cell culture, HMI-1a3 reduced cell viability or proliferation in all cell lines as determined by the metabolic activity of the cells (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay) and thymidine incorporation. However, the mechanism of action in LNCaP cells was different to that in DU145 or PC3 cells. In LNCaP cells, HMI-1a3 induced a PKC-dependent activation of caspase 3/7, indicating an apoptotic response, whereas in DU145 and PC3 cells, it induced senescence, which was independent of PKC. This was observed as typical senescent morphology, increased beta-galactosidase activity, and upregulation of the senescence marker p21 and downregulation of E2F transcription factor 1. Using a multicellular spheroid model, we further showed that HMI-1a3 affects the growth of LNCaP and DU145 cells in a 3D culture, emphasizing its potential as a lead compound for cancer drug development.
  • Lindevall, Mari (Helsingin yliopisto, 2021)
    The purpose of this systematic review is to investigate the usage of artificial intelligence in the pharmaceutical industry in the fields of pharmaceutical manufacturing, product development, and quality control. Today, developing and getting a new drug on the market is time-consuming, ineffective, and expensive. Artificial intelligence is seen as one possible solution to the problems of the pharmaceutical industry. From 734 articles 77 academic study articles were included. Included articles showed artificial neural networks to be the most used artificial intelligence method between 1991 and 2021. The search was conducted from three databases with the following inclusion criteria: studies using AI in either pharmaceutical manufacturing, product development or quality control, English as the language, and Western medicine-based pharmacy as a branch of science. This systematic literature review has three main limitations: the possibility of an important search word missing from the search algorithm, the selection of articles according to one person's assessment, and the possible narrow picture of the used artificial intelligence methods in the pharmaceutical industry, as pharmaceutical companies also research the subject. The use of artificial intelligence in product development has been studied the most, while its use in quality control has been studied the least. In the studies, tablets were a popular drug form, while biological drugs were underrepresented. In total, the number of studies published increased over three decades. However, most of the articles were published in 2020. Nearly half of the articles had some connection to a pharmaceutical company, indicating the interest of both the academy and pharmaceutical companies in the use of artificial intelligence in manufacturing, product development, and quality control. In the future, the efficacy of artificial intelligence, as well as its limitations as a method, should be investigated to conclude its potential to play a key role in reforming the pharmaceutical industry. The results of the study show that a wave of artificial intelligence has arrived in the pharmaceutical industry, however, its real benefits will only be seen with future research.
  • Dauvilliers, Yves; Bogan, Richard K.; Sonka, Karel; Partinen, Markku; Foldvary-Schaefer, Nancy; Thorpy, Michael J. (2022)
    Lower-sodium oxybate (LXB) is an oxybate medication approved to treat cataplexy or excessive daytime sleepiness (EDS) in patients with narcolepsy 7 years of age and older in the United States. LXB was developed as an alternative to sodium oxybate (SXB), because the incidence of cardiovascular comorbidities is higher in patients with narcolepsy and there is an elevated cardiovascular risk associated with high sodium consumption. LXB has a unique formulation of calcium, magnesium, potassium, and sodium ions, containing 92% less sodium than SXB. Whereas the active oxybate moiety is the same for LXB and SXB, their pharmacokinetic profiles are not bioequivalent; therefore, a phase 3 trial in participants with narcolepsy was conducted for LXB. This review summarizes the background on oxybate as a therapeutic agent and its potential mechanism of action on the gammaaminobutyric acid type B (GABA B ) receptor at noradrenergic and dopaminergic neurons, as well as at thalamocortical neurons. The rationale leading to the development of LXB as a lower-sodium alternative to SXB and the key efficacy and safety data supporting its approval for both adult and pediatric patients with narcolepsy are also discussed. LXB was approved in August 2021 in the United States for the treatment of idiopathic hypersomnia in adults. Potential future developments in the field of oxybate medications may include novel formulations and expanded indications for other diseases.
  • Chang, Mingyang; Bogacheva, Mariia S.; Lou, Yan-Ru (2021)
    The current organoid culture systems allow pluripotent and adult stem cells to self-organize to form three-dimensional (3D) structures that provide a faithful recapitulation of the architecture and function of in vivo organs. In particular, human pluripotent stem cell-derived liver organoids (PSC-LOs) can be used in regenerative medicine and preclinical applications, such as disease modeling and drug discovery. New bioengineering tools, such as microfluidics, biomaterial scaffolds, and 3D bioprinting, are combined with organoid technologies to increase the efficiency of hepatic differentiation and enhance the functional maturity of human PSC-LOs by precise control of cellular microenvironment. Long-term stabilization of hepatocellular functions of in vitro liver organoids requires the combination of hepatic endodermal, endothelial, and mesenchymal cells. To improve the biological function and scalability of human PSC-LOs, bioengineering methods have been used to identify diverse and zonal hepatocyte populations in liver organoids for capturing heterogeneous pathologies. Therefore, constructing engineered liver organoids generated from human PSCs will be an extremely versatile tool in in vitro disease models and regenerative medicine in future. In this review, we aim to discuss the recent advances in bioengineering technologies in liver organoid culture systems that provide a timely and necessary study to model disease pathology and support drug discovery in vitro and to generate cell therapy products for transplantation.
  • BIOS Consortium; Reese, Sarah E.; Xu, Cheng-Jian; den Dekker, Herman T.; Lahti, Jari; Kajantie, Eero; Kere, Juha; Räikkönen, Katri (2019)
    Background: Epigenetic mechanisms, including methylation, can contribute to childhood asthma. Identifying DNA methylation profiles in asthmatic patients can inform disease pathogenesis. Objective: We sought to identify differential DNA methylation in newborns and children related to childhood asthma. Methods: Within the Pregnancy And Childhood Epigenetics consortium, we performed epigenome-wide meta-analyses of school-age asthma in relation to CpG methylation (Illumina450K) in blood measured either in newborns, in prospective analyses, or cross-sectionally in school-aged children. We also identified differentially methylated regions. Results: In newborns (8 cohorts, 668 cases), 9 CpGs (and 35 regions) were differentially methylated (epigenome-wide significance, false discovery rate <0.05) in relation to asthma development. In a cross-sectional meta-analysis of asthma and methylation in children (9 cohorts, 631 cases), we identified 179 CpGs (false discovery rate <0.05) and 36 differentially methylated regions. In replication studies of methylation in other tissues, most of the 179 CpGs discovered in blood replicated, despite smaller sample sizes, in studies of nasal respiratory epithelium or eosinophils. Pathway analyses highlighted enrichment for asthma-relevant immune processes and overlap in pathways enriched both in newborns and children. Gene expression correlated with methylation at most loci. Functional annotation supports a regulatory effect on gene expression at many asthma-associated CpGs. Several implicated genes are targets for approved or experimental drugs, including IL5RA and KCNH2. Conclusion: Novel loci differentially methylated in newborns represent potential biomarkers of risk of asthma by school age. Cross-sectional associations in children can reflect both risk for and effects of disease. Asthma-related differential methylation in blood in children was substantially replicated in eosinophils and respiratory epithelium.
  • Sokka, Iris Katariina; Ekholm, Filip S.; Johansson, Mikael P. (2019)
    Monomethyl auristatin E and monomethyl auristatin F are widely used cytotoxic agents in antibody-drug conjugates (ADCs), a group of promising cancer drugs. The ADCs specifically target cancer cells, releasing the auristatins inside, which results in the prevention of mitosis. The auristatins suffer from a potentially serious flaw, however. In solution, the molecules exist in an equal mixture of two conformers, cis and trans. Only the trans-isomer is biologically active and the isomerization process, i.e., the conversion of cis to trans is slow. This significantly diminishes the efficiency of the drugs and their corresponding ADCs, and perhaps more importantly, raises concerns over drug safety. The potency of the auristatins would be enhanced by decreasing the amount of the biologically inactive isomer, either by stabilizing the transisomer or destabilizing the cis-isomer. Here, we follow the computer-aided design strategy of shifting the conformational equilibrium and employ high-level quantum chemical modeling to identify promising candidates for improved auristatins. Coupled cluster calculations predict that a simple halogenation in the norephedrine/phenylalanine residues shifts the isomer equilibrium almost completely toward the active trans-conformation, due to enhanced intramolecular interactions specific to the active isomer.
  • Kenttä, Laura (Helsingfors universitet, 2015)
    Susceptibility to antibiotics is constantly developing in bacteria due to selection pressure caused by use of antibiotics. For this reason, finding new antimicrobial substances is imperative. High-throughput screening (HTS) is an important tool to find new active substances. The need to analyse as many substances in as small time as possible is emphasised in modern drug development. Robust methods, suitable for fast throughput of substances, miniaturisation and automation, are particularly useful. In the context of antimicrobial screening, methods utilising bioluminescence can correspond this need, and genetic engineering can help in developing bacterial strains with beneficial features for screening. In this work, two screening methods were developed and optimised using genetically engineered Escherichia coli strains. The screening methods make use of the bioluminescent properties of the strains, and the methods can be used to screen compound libraries for antimicrobials rapidly enough to approach HTS. The strain E. coli WZM120/pCGLS 11 is constitutively luminescent, so weakening of luminescence means the cell viability weakens. The strain E. coli K12/pCSS305, where luminescence is produced by a heat-inducible runaway plasmid, can be used to especially detect compounds inhibiting DNA replication. In developing the method, workflow was optimised and conditions were validated so as to enable possible HTS campaigns. The target was to create as simple, fast and reproducible a method as possible. The Z' values calculated in assessing the performance are excellent for a cell-based method. The signal is readily distinguishable, the bacterial strains are in a stable manner, and the method is well reproducible. It is possible to continue assay development from 96-well format to 384-well format.
  • Wurtz, Peter; Wang, Qin; Soininen, Pasi; Kangas, Antti J.; Fatemifar, Ghazaleh; Tynkkynen, Tuulia; Tiainen, Mika; Perola, Markus; Tillin, Therese; Hughes, Alun D.; Mantyselka, Pekka; Kahonen, Mika; Lehtimaki, Terho; Sattar, Naveed; Hingorani, Aroon D.; Casas, Juan-Pablo; Salomaa, Veikko; Kivimaki, Mika; Jarvelin, Marjo-Riitta; Smith, George Davey; Vanhala, Mauno; Lawlor, Debbie A.; Raitakari, Olli T.; Chaturvedi, Nish; Kettunen, Johannes; Ala-Korpela, Mika (2016)
    BACKGROUND Statins are first-line therapy for cardiovascular disease prevention, but their systemic effects across lipoprotein subclasses, fatty acids, and circulating metabolites remain incompletely characterized. OBJECTIVES This study sought to determine the molecular effects of statin therapy on multiple metabolic pathways. METHODS Metabolic profiles based on serum nuclear magnetic resonance metabolomics were quantified at 2 time points in 4 population-based cohorts from the United Kingdom and Finland (N = 5,590; 2.5 to 23.0 years of follow-up). Concentration changes in 80 lipid and metabolite measures during follow-up were compared between 716 individuals who started statin therapy and 4,874 persistent nonusers. To further understand the pharmacological effects of statins, we used Mendelian randomization to assess associations of a genetic variant known to mimic inhibition of HMG-CoA reductase (the intended drug target) with the same lipids and metabolites for 27,914 individuals from 8 population-based cohorts. RESULTS Starting statin therapy was associated with numerous lipoprotein and fatty acid changes, including substantial lowering of remnant cholesterol (80% relative to low-density lipoprotein cholesterol [LDL-C]), but only modest lowering of triglycerides (25% relative to LDL-C). Among fatty acids, omega-6 levels decreased the most (68% relative to LDL-C); other fatty acids were only modestly affected. No robust changes were observed for circulating amino acids, ketones, or glycolysis-related metabolites. The intricate metabolic changes associated with statin use closely matched the association pattern with rs12916 in the HMGCR gene (R-2 = 0.94, slope 1.00 +/- 0.03). CONCLUSIONS Statin use leads to extensive lipid changes beyond LDL-C and appears efficacious for lowering remnant cholesterol. Metabolomic profiling, however, suggested minimal effects on amino acids. The results exemplify how detailed metabolic characterization of genetic proxies for drug targets can inform indications, pleiotropic effects, and pharmacological mechanisms. (C) 2016 by the American College of Cardiology Foundation.
  • Siddiqui, Farid Ahmad; Parkkola, Hanna; Vukic, Vladimir; Oetken-Lindholm, Christina; Jaiswal, Alok; Kiriazis, Alexandros; Pavic, Karolina; Aittokallio, Tero; Salminen, Tiina A.; Abankwa, Daniel (2021)
    Simple Summary The correct folding of proteins is essential for their activity. Therefore, cells have evolved protein-folding chaperones, such as Hsp90. Interestingly, in several cancer cells, Hsp90 appears to have a role that is more important than normal. The current working model suggests that, with the help of its co-chaperone, Cdc37, it stabilizes mutant kinases. However, Hsp90, together with Cdc37, assists additional proteins that may be relevant in cancer. We demonstrate that the Hsp90-dependent stability of the transcription factor HIF-1 alpha and one of its downstream transcriptional targets, galectin-3, is important to maintain the elevated activity of the major oncogene KRAS. This is because galectin-3 stabilizes the MAPK-signaling complexes of K-Ras, which is called a nanocluster. In addition, we identified six drug-like small molecules that inhibit the Hsp90/Cdc37 protein interface at low micro molar concentrations. Given the co-occurrence of mutant KRAS with high HIF-1 alpha and high galectin-3 levels in pancreatic cancer, our results suggest an application of Hsp90 inhibitors in this cancer type. The ATP-competitive inhibitors of Hsp90 have been tested predominantly in kinase addicted cancers; however, they have had limited success. A mechanistic connection between Hsp90 and oncogenic K-Ras is not known. Here, we show that K-Ras selectivity is enabled by the loss of the K-Ras membrane nanocluster modulator galectin-3 downstream of the Hsp90 client HIF-1 alpha. This mechanism suggests a higher drug sensitivity in the context of KRAS mutant, HIF-1 alpha-high and/or Gal3-high cancer cells, such as those found, in particular, in pancreatic adenocarcinoma. The low toxicity of conglobatin further indicates a beneficial on-target toxicity profile for Hsp90/Cdc37 interface inhibitors. We therefore computationally screened >7 M compounds, and identified four novel small molecules with activities of 4 mu M-44 mu M in vitro. All of the compounds were K-Ras selective, and potently decreased the Hsp90 client protein levels without inducing the heat shock response. Moreover, they all inhibited the 2D proliferation of breast, pancreatic, and lung cancer cell lines. The most active compounds from each scaffold, furthermore, significantly blocked 3D spheroids and the growth of K-Ras-dependent microtumors. We foresee new opportunities for improved Hsp90/Cdc37 interface inhibitors in cancer and other aging-associated diseases.
  • Janssens, Rosanne; Lang, Tamika; Vallejo, Ana; Galinsky, Jayne; Plate, Ananda; Morgan, Kate; Cabezudo, Elena; Silvennoinen, Raija; Coriu, Daniel; Badelita, Sorina; Irimia, Ruxandra; Anttonen, Minna; Manninen, Riikka-Leena; Schoefs, Elise; Vandebroek, Martina; Vanhellemont, Anneleen; Delforge, Michel; Stevens, Hilde; Simoens, Steven; Huys, Isabelle (2021)
    Background: Investigational and marketed drugs for the treatment of multiple myeloma (MM) are associated with a range of characteristics and uncertainties regarding long term side-effects and efficacy. This raises questions about what matters most to patients living with this disease. This study aimed to understand which characteristics MM patients find most important, and hence should be included as attributes and levels in a subsequent quantitative preference survey among MM patients. Methods: This qualitative study involved: (i) a scoping literature review, (ii) discussions with MM patients (n = 24) in Belgium, Finland, Romania, and Spain using Nominal Group Technique, (iii) a qualitative thematic analysis including multi-stakeholder discussions. Results: MM patients voiced significant expectations and hopes that treatments would extend their lives and reduce their cancer signs and symptoms. Participants however raised concerns about life-threatening side-effects that could cause permanent organ damage. Bone fractures and debilitating neuropathic effects (such as chronic tingling sensations) were highlighted as major issues reducing patients' independence and mobility. Patients discussed the negative impact of the following symptoms and side-effects on their daily activities: thinking problems, increased susceptibility to infections, reduced energy, pain, emotional problems, and vision problems. MM patients were concerned with uncertainties regarding the durability of positive treatment outcomes, and the cause, severity, and duration of their symptoms and side-effects. Patients feared short-term positive treatment responses complicated by permanent, severe side-effects and symptoms. Conclusions: This study gained an in-depth understanding of the treatment and disease-related characteristics and types of attribute levels (severity, duration) that are most important to MM patients. Results from this study argue in favor of MM drug development and individual treatment decision-making that focuses not only on extending patients' lives but also on addressing those symptoms and side-effects that significantly impact MM patients' quality of life. This study underscores a need for transparent communication toward MM patients about MM treatment outcomes and uncertainties regarding their long-term efficacy and safety. Finally, this study may help drug developers and decision-makers understand which treatment outcomes and uncertainties are most important to MM patients and therefore should be incorporated in MM drug development, evaluation, and clinical practice.
  • Hämäläinen, Iiro; Törnwall, Outi; Simell, Birgit; Zatloukal, Kurt; Perola, Markus; van Ommen, Gert-Jan B. (2019)
    Public-private partnerships (PPP) are an efficient means to advance scientific discoveries and boost the medical innovations needed to improve precision medicine. The increasing number and novel nature of such collaborations is keeping the biomedical field in a constant flux. Here we provide an update on PPP development involving academic biobanks in the BBMRI community (the European Biobanking and BioMolecular Resources Research Infrastructure) and report the views on PPP of 20 key players from this field. The interviewed academic representants broadly show interest for their institution to establish PPP and initiate or partner with BBMRI expert centers. The results indicate that PPP has gained foothold in this area of biomedical research, with great promise to facilitate access to samples and data and to improve data interoperability and reproducibility.
  • Yli-Rantala, Anni (Helsingfors universitet, 2014)
    Zebrafish (Danio rerio) is a vertebrate model organism. It is suited for many phases of drug development process like toxicological studies. The major advantage of using zebrafish is the possibility to conduct high-throughput screens on a whole vertebrate animal. However, there is not as much knowledge about zebrafish as there is about other model organisms. Therefore there might be differences between zebrafish and humans that affect the use of zebrafish as a model in the drug development process. The purpose of this thesis was to characterize the structure of the zebrafish oxytocin system and assess the role of oxytocin on zebrafish behaviour. In humans defects in the oxytocin system have been linked to many psychiatric disorders like autism. If the mammalian and zebrafish oxytocin systems resembled each other functionally and structurally, it would enable the use of zebrafish as a model when studying the role of oxytocin in pathophysiology of diseases and also in oxytocin system related drug development. The structure and development of zebrafish oxytocin system was studied by staining adult zebrafish brain cryosections and larval brains with antibodies made against mammalian oxytocin. The specificity of the antibodies to recognize zebrafish oxytocin was determined by absorption and cross-reactivity controls. The role of oxytocin on zebrafish locomotion was studied by inhibiting the splicing of oxytocin messenger RNA with morpholino oligonucleotides (MOs). The MOs were used to address the relevance of the model in pharmacology, since the zebrafish oxytocin receptors have not been expressed and pharmacologically characterized. In zebrafish oxytocin was produced in the cells of the preoptic nucleus. There were thick oxytocin fibers towards the pituitary and also thinner fibers into areas in the telencephalon, diencephalon, mesencephalon and rhombencephalon. One of the MOs was able to inhibit the production of oxytocin with a dose that did not cause morphological abnormalities. The MO reduced the locomotor activity of the fish, but the specificity of the MO has to be determined. The structure of the zebrafish oxytocin system resembles mammalian oxytocin system in terms of the location of oxytocin cells and fiber projections. Therefore zebrafish seems a suitable model organism for oxytocin research. However, the structure of the zebrafish oxytocin receptor system and the effect of oxytocin on other behavioural aspects have to be determined in order to further evaluate the applicability of zebrafish for oxytocin research.