17β-hydroxysteroid dehydrogenase type 10 (17β-HSD10) is a multifunctional mitochondrial enzyme and putative drug target for the treatment of various pathologies including Alzheimer's disease or some types of hormone-dependent cancer. In this study, a series of new benzothiazolylurea-based inhibitors were developed based on the structure-activity relationship (SAR) study of previously published compounds and predictions of their physico-chemical properties. This led to the identification of several submicromolar inhibitors (IC50 ∼0.3 μM), the most potent compounds within the benzothiazolylurea class known to date. The positive interaction with 17β-HSD10 was further confirmed by differential scanning fluorimetry and the best molecules were found to be cell penetrable. In addition, the best compounds weren't found to have additional effects for mitochondrial off-targets and cytotoxic or neurotoxic effects. The two most potent inhibitors 9 and 11 were selected for in vivo pharmacokinetic study after intravenous and peroral administration. Although the pharmacokinetic results were not fully conclusive, it seemed that compound 9 was bioavailable after peroral administration and could penetrate into the brain (brain-plasma ratio 0.56).

• Klíčová slova
• 17β-hydroxysteroid dehydrogenase type 10 (17β-HSD10), Alzheimer's disease, Amyloid-binding alcohol dehydrogenase (ABAD), Enzyme inhibition, Pharmacokinetics,
• MeSH
• 17-hydroxysteroidní dehydrogenasy MeSH
• Alzheimerova nemoc * farmakoterapie   MeSH
• inhibitory enzymů chemie   MeSH
• lidé MeSH
• mozek metabolismus   MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 17-hydroxysteroidní dehydrogenasy MeSH
• 3 (or 17)-beta-hydroxysteroid dehydrogenase MeSH Prohlížeč
• inhibitory enzymů MeSH

Mitochondrial enzymes are targets of newly synthesized drugs being tested for the treatment of neurodegenerative disorders, such as Alzheimer's disease (AD). The enzyme 17β-hydroxysteroid dehydrogenase type 10 (HSD10) is a multifunctional mitochondrial protein that is thought to play a role in the pathophysiology of AD and is one of the targets of new potential AD drugs. The in vitro effects of frentizole, riluzole, AG18051, and 42 novel modulators of HSD10 (potential AD drugs) on citrate synthase (CS) activity, monoamine oxidase (MAO) activity, complex I- or complex II-linked mitochondrial respiratory rate, and complex I activity were measured in isolated pig brain mitochondria. Based on their minimal inhibitory effects on the respiratory rate of mitochondria and CS and complex I activity, six novel compounds were selected for further testing. Assuming that inhibition of MAO-B could be a desirable effect of AD drugs, only AG18051 and one new compound met the criteria for MAO-B inhibition with minimal drug-induced effects on mitochondrial respiration. In conclusion, our in vitro screening of mitochondrial effect of novel potential AD drugs has enabled the selection of the most promising molecules for further testing that are relatively safe in terms of drug-induced mitochondrial toxicity.

• Klíčová slova
• 17β-hydroxysteroid dehydrogenase type 10, Alzheimer’s disease, Drug development, Electron transport chain, Mitochondrial toxicity, Monoamine oxidase,
• MeSH
• 17-hydroxysteroidní dehydrogenasy antagonisté a inhibitory   toxicita   MeSH
• buněčné dýchání účinky léků   MeSH
• inhibitory enzymů terapeutické užití   toxicita   MeSH
• lidé MeSH
• mitochondrie účinky léků   MeSH
• modely u zvířat MeSH
• neurodegenerativní nemoci farmakoterapie   MeSH
• prasata MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 17-hydroxysteroidní dehydrogenasy MeSH
• inhibitory enzymů MeSH

Midostaurin is an FMS-like tyrosine kinase 3 receptor (FLT3) inhibitor that provides renewed hope for treating acute myeloid leukaemia (AML). The limited efficacy of this compound as a monotherapy contrasts with that of its synergistic combination with standard cytarabine and daunorubicin (Dau), suggesting a therapeutic benefit that is not driven only by FLT3 inhibition. In an AML context, the activity of the enzyme aldo-keto reductase 1C3 (AKR1C3) is a crucial factor in chemotherapy resistance, as it mediates the intracellular transformation of anthracyclines to less active hydroxy metabolites. Here, we report that midostaurin is a potent inhibitor of Dau inactivation mediated by AKR1C3 in both its recombinant form as well as during its overexpression in a transfected cell model. Likewise, in the FLT3- AML cell line KG1a, midostaurin was able to increase the cellular accumulation of Dau and significantly decrease its metabolism by AKR1C3 simultaneously. The combination of those mechanisms increased the nuclear localization of Dau, thus synergizing its cytotoxic effects on KG1a cells. Our results provide new in vitro evidence of how the therapeutic activity of midostaurin could operate beyond targeting the FLT3 receptor.

• Klíčová slova
• AKR1C3, AML therapy, Anthracyclines, Midostaurin, Multidrug resistance,
• MeSH
• akutní myeloidní leukemie farmakoterapie   enzymologie   genetika   patologie   MeSH
• biotransformace MeSH
• daunomycin metabolismus   farmakologie   MeSH
• HCT116 buňky MeSH
• inhibitory enzymů farmakologie   MeSH
• kolorektální nádory farmakoterapie   enzymologie   genetika   patologie   MeSH
• lidé MeSH
• protein AKR1C3 antagonisté a inhibitory   genetika   metabolismus   MeSH
• protokoly protinádorové kombinované chemoterapie farmakologie   MeSH
• staurosporin analogy a deriváty   farmakologie   MeSH
• synergismus léků MeSH
• tyrosinkinasa 3 podobná fms antagonisté a inhibitory   genetika   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• AKR1C3 protein, human MeSH Prohlížeč
• daunomycin MeSH
• FLT3 protein, human MeSH Prohlížeč
• inhibitory enzymů MeSH
• midostaurin MeSH Prohlížeč
• protein AKR1C3 MeSH
• staurosporin MeSH
• tyrosinkinasa 3 podobná fms MeSH

In early stages of Alzheimer's disease (AD), amyloid beta (Aβ) accumulates in the mitochondrial matrix and interacts with mitochondrial proteins, such as cyclophilin D (cypD) and 17β-hydroxysteroid dehydrogenase 10 (17β-HSD10). Multiple processes associated with AD such as increased production or oligomerization of Aβ affect these interactions and disbalance the equilibrium between the biomolecules, which contributes to mitochondrial dysfunction. Here, we investigate the effect of the ionic environment on the interactions of Aβ (Aβ1-40, Aβ1-42) with cypD and 17β-HSD10 using a surface plasmon resonance (SPR) biosensor. We show that changes in concentrations of K+ and Mg2+ significantly affect the interactions and may increase the binding efficiency between the biomolecules by up to 35% and 65% for the interactions with Aβ1-40 and Aβ1-42, respectively, in comparison with the physiological state. We also demonstrate that while the binding of Aβ1-40 to cypD and 17β-HSD10 takes place preferentially around the physiological concentrations of ions, decreased concentrations of K+ and increased concentrations of Mg2+ promote the interaction of both mitochondrial proteins with Aβ1-42. These results suggest that the ionic environment represents an important factor that should be considered in the investigation of biomolecular interactions taking place in the mitochondrial matrix under physiological as well as AD-associated conditions.

• Klíčová slova
• 17β-hydroxysteroid dehydrogenase 10 (17β-HSD10), amyloid beta (Aβ), biomolecular interactions, cyclophilin D (cypD), ionic environment, mitochondrial matrix, surface plasmon resonance (SPR),
• MeSH
• 17-hydroxysteroidní dehydrogenasy chemie   genetika   MeSH
• Alzheimerova nemoc diagnóza   genetika   patologie   MeSH
• amyloidní beta-protein chemie   MeSH
• biosenzitivní techniky metody   MeSH
• ionty chemie   MeSH
• lidé MeSH
• mitochondriální proteiny chemie   MeSH
• mitochondrie chemie   MeSH
• peptidové fragmenty chemie   genetika   MeSH
• peptidylprolylisomerasa F chemie   genetika   MeSH
• povrchová plasmonová rezonance metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 17-hydroxysteroidní dehydrogenasy MeSH
• 3 (or 17)-beta-hydroxysteroid dehydrogenase MeSH Prohlížeč
• amyloidní beta-protein MeSH
• ionty MeSH
• mitochondriální proteiny MeSH
• peptidové fragmenty MeSH
• peptidylprolylisomerasa F MeSH

Progressive mitochondrial dysfunction due to the accumulation of amyloid beta (Aβ) peptide within the mitochondrial matrix represents one of the key characteristics of Alzheimer's disease (AD) and appears already in its early stages. Inside the mitochondria, Aβ interacts with a number of biomolecules, including cyclophilin D (cypD) and 17β-hydroxysteroid dehydrogenase type 10 (17β-HSD10), and affects their physiological functions. However, despite intensive ongoing research, the exact mechanisms through which Aβ impairs mitochondrial functions remain to be explained. In this work, we studied the interactions of Aβ with cypD and 17β-HSD10 in vitro using the surface plasmon resonance (SPR) method and determined the kinetic parameters (association and dissociation rates) of these interactions. This is the first work which determines all these parameters under the same conditions, thus, enabling direct comparison of relative affinities of Aβ to its mitochondrial binding partners. Moreover, we used the determined characteristics of the individual interactions to simulate the concurrent interactions of Aβ with cypD and 17β-HSD10 in different model situations associated with the progression of AD. This study not only advances the understanding of Aβ-induced processes in mitochondria during AD, but it also provides a new perspective on research into complex multi-interaction biomolecular processes in general.

• Klíčová slova
• 17β-hydroxysteroid dehydrogenase 10 (17β-HSD10), amyloid beta (Aβ), biomolecular interaction analysis, cyclophilin D (cypD), kinetic parameters, surface plasmon resonance (SPR),
• MeSH
• 17-hydroxysteroidní dehydrogenasy chemie   metabolismus   MeSH
• Alzheimerova nemoc metabolismus   MeSH
• amyloidní beta-protein chemie   metabolismus   MeSH
• biosenzitivní techniky MeSH
• lidé MeSH
• mitochondriální proteiny chemie   metabolismus   MeSH
• peptidylprolylisomerasa F chemie   metabolismus   MeSH
• povrchová plasmonová rezonance MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 17-hydroxysteroidní dehydrogenasy MeSH
• amyloidní beta-protein MeSH
• mitochondriální proteiny MeSH
• peptidylprolylisomerasa F MeSH

The nucleus-encoded 17β-hydroxysteroid dehydrogenase type 10 (17β-HSD10) regulates cyclophilin D (cypD) in the mitochondrial matrix. CypD regulates opening of mitochondrial permeability transition pores. Both mechanisms may be affected by amyloid β peptides accumulated in mitochondria in Alzheimer's disease (AD). In order to clarify changes occurring in brain mitochondria, we evaluated interactions of both mitochondrial proteins in vitro (by surface plasmon resonance biosensor) and detected levels of various complexes of 17β-HSD10 formed in vivo (by sandwich ELISA) in brain mitochondria isolated from the transgenic animal model of AD (homozygous McGill-R-Thy1-APP rats) and in cerebrospinal fluid samples of AD patients. By surface plasmon resonance biosensor, we observed the interaction of 17β-HSD10 and cypD in a direct real-time manner and determined, for the first time, the kinetic parameters of the interaction (ka 2.0 × 105 M1s-1, kd 5.8 × 104 s-1, and KD 3.5 × 10-10 M). In McGill-R-Thy1-APP rats compared to controls, levels of 17β-HSD10-cypD complexes were decreased and those of total amyloid β increased. Moreover, the levels of 17β-HSD10-cypD complexes were decreased in cerebrospinal fluid of individuals with AD (in mild cognitive impairment as well as dementia stages) or with Frontotemporal lobar degeneration (FTLD) compared to cognitively normal controls (the sensitivity of the complexes to AD dementia was 92.9%, that to FTLD 73.8%, the specificity to AD dementia equaled 91.7% in a comparison with the controls but only 26.2% with FTLD). Our results demonstrate the weakened ability of 17β-HSD10 to regulate cypD in the mitochondrial matrix probably via direct effects of amyloid β. Levels of 17β-HSD10-cypD complexes in cerebrospinal fluid seem to be the very sensitive indicator of mitochondrial dysfunction observed in neurodegeneration but unfortunately not specific to AD pathology. We do not recommend it as the new biomarker of AD.

• Klíčová slova
• Alzheimer's disease, Amyloid β, Cerebrospinal fluid, Frontotemporal lobar degeneration, Mitochondrial matrix proteins, Transgenic rat model,
• MeSH
• 17-hydroxysteroidní dehydrogenasy mozkomíšní mok   metabolismus   MeSH
• Alzheimerova nemoc metabolismus   MeSH
• amyloidový prekurzorový protein beta genetika   MeSH
• kinetika MeSH
• lidé MeSH
• mitochondrie metabolismus   MeSH
• mozek metabolismus   MeSH
• peptidylprolylisomerasa F metabolismus   MeSH
• potkani transgenní MeSH
• potkani Wistar MeSH
• povrchová plasmonová rezonance MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 17-hydroxysteroidní dehydrogenasy MeSH
• amyloidový prekurzorový protein beta MeSH
• peptidylprolylisomerasa F MeSH

Metabolic reprogramming of tumor cells involves upregulation of fatty acid (FA) synthesis to support high bioenergetic demands and membrane synthesis. This has been shown for cytosolic synthesis of FAs with up to 16 carbon atoms. Synthesis of long-chain fatty acids (LCFAs), including ω-6 and ω-3 polyunsaturated FAs, takes place at the endoplasmic reticulum. Despite increasing evidence for an important role of LCFAs in cancer, the impact of their synthesis in cancer cell growth has scarcely been studied. Here, we demonstrated that silencing of 17β-hydroxysteroid dehydrogenase type 12 (17β-HSD12), essentially catalyzing the 3-ketoacyl-CoA reduction step in LCFA production, modulates proliferation and migration of breast cancer cells in a cell line-dependent manner. Increased proliferation and migration after 17β-HSD12 knockdown were partly mediated by metabolism of arachidonic acid towards COX2 and CYP1B1-derived eicosanoids. Decreased proliferation was rescued by increased glucose concentration and was preceded by reduced ATP production through oxidative phosphorylation and spare respiratory capacity. In addition, 17β-HSD12 silencing was accompanied by alterations in unfolded protein response, including a decrease in CHOP expression and increase in eIF2α activation and the folding chaperone ERp44. Our study highlights the significance of LCFA biosynthesis for tumor cell physiology and unveils unknown aspects of breast cancer cell heterogeneity.

• Klíčová slova
• 17β-Hydroxysteroid dehydrogenase, Biosynthesis, Cancer, Endoplasmic reticulum, Long-chain fatty acid, Unfolded protein response,
• MeSH
• 17-hydroxysteroidní dehydrogenasy genetika   metabolismus   MeSH
• acylkoenzym A metabolismus   MeSH
• lidé MeSH
• lipogeneze MeSH
• mastné kyseliny metabolismus   MeSH
• MFC-7 buňky MeSH
• nádorové buněčné linie MeSH
• nádory prsu genetika   metabolismus   patologie   MeSH
• pohyb buněk MeSH
• proliferace buněk MeSH
• umlčování genů MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 17-hydroxysteroidní dehydrogenasy MeSH
• 17beta-hydroxysteroid dehydrogenase type 3 MeSH Prohlížeč
• acylkoenzym A MeSH
• mastné kyseliny MeSH

In early stages of Alzheimer's disease (AD), amyloid-β (Aβ) accumulates in neuronal mitochondria where it interacts with a number of biomolecules including 17beta-hydroxysteroide dehydrogenase 10 (17β-HSD10) and cyclophilin D (cypD). It has been hypothesized that 17β-HSD10 interacts with cypD preventing it from opening mitochondrial permeability transition pores and that its regulation during AD may be affected by the accumulation of Aβ. In this work, we demonstrate for the first time that 17β-HSD10 and cypD form a stable complex in vitro. Furthermore, we show that factors, such as pH, ionic environment and the presence of Aβ, affect the ability of 17β-HSD10 to bind cypD. We demonstrate that K+ and Mg2+ ions present at low levels may facilitate this binding. We also show that different fragments of Aβ (Aβ1-40 and Aβ1-42) affect the interaction between 17β-HSD10 and cypD differently and that Aβ1-42 (in contrast to Aβ1-40) is capable of simultaneously binding both 17β-HSD10 and cypD in a tri-complex.

• MeSH
• 17-hydroxysteroidní dehydrogenasy metabolismus   MeSH
• Alzheimerova nemoc metabolismus   patologie   MeSH
• amyloidní beta-protein metabolismus   MeSH
• lidé MeSH
• mitochondrie metabolismus   MeSH
• peptidylprolylisomerasa F metabolismus   MeSH
• přechodový pór mitochondriální permeability MeSH
• techniky in vitro MeSH
• transportní proteiny mitochondriální membrány metabolismus   MeSH
• vápník metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 17-hydroxysteroidní dehydrogenasy MeSH
• 3 (or 17)-beta-hydroxysteroid dehydrogenase MeSH Prohlížeč
• amyloidní beta-protein MeSH
• peptidylprolylisomerasa F MeSH
• přechodový pór mitochondriální permeability MeSH
• transportní proteiny mitochondriální membrány MeSH
• vápník MeSH

: It has long been established that mitochondrial dysfunction in Alzheimer's disease (AD) patients can trigger pathological changes in cell metabolism by altering metabolic enzymes such as the mitochondrial 17β-hydroxysteroid dehydrogenase type 10 (17β-HSD10), also known as amyloid-binding alcohol dehydrogenase (ABAD). We and others have shown that frentizole and riluzole derivatives can inhibit 17β-HSD10 and that this inhibition is beneficial and holds therapeutic merit for the treatment of AD. Here we evaluate several novel series based on benzothiazolylurea scaffold evaluating key structural and activity relationships required for the inhibition of 17β-HSD10. Results show that the most promising of these compounds have markedly increased potency on our previously published inhibitors, with the most promising exhibiting advantageous features like low cytotoxicity and target engagement in living cells.

• Klíčová slova
• 17β-hydroxysteroid dehydrogenase type 10 (17β-HSD10), amyloid binding alcohol dehydrogenase (ABAD), benzothiazole, Alzheimer’s disease (AD), amyloid-beta peptide (Aβ), mitochondria,
• MeSH
• 17-hydroxysteroidní dehydrogenasy antagonisté a inhibitory   chemie   MeSH
• Alzheimerova nemoc farmakoterapie   MeSH
• amyloidní beta-protein metabolismus   MeSH
• benzothiazoly chemie   MeSH
• buněčné linie MeSH
• lidé MeSH
• mitochondrie metabolismus   MeSH
• močovina chemie   MeSH
• molekulární struktura MeSH
• racionální návrh léčiv MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 17-hydroxysteroidní dehydrogenasy MeSH
• amyloidní beta-protein MeSH
• benzothiazoly MeSH
• močovina MeSH

Buparlisib is a pan-class I phosphoinositide 3-kinase (PI3K) inhibitor and is currently under clinical evaluation for the treatment of different cancers. Because PI3K signalling is related to cell proliferation and resistance to chemotherapy, new therapeutic approaches are focused on combining PI3K inhibitors with other anti-cancer therapeutics. Carbonyl-reducing enzymes catalyse metabolic detoxification of anthracyclines and reduce their cytotoxicity. In the present work, the effects of buparlisib were tested on six human recombinant carbonyl-reducing enzymes: AKR1A1, AKR1B1, AKR1B10, AKR1C3, and AKR7A2 from the aldo-keto reductase superfamily and CBR1 from the short-chain dehydrogenase/reductase superfamily, all of which participate in the metabolism of daunorubicin. Buparlisib exhibited the strongest inhibitory effect on recombinant AKR1C3, with a half-maximal inhibitory concentration (IC50) of 9.5 μM. Its inhibition constant Ki was found to be 14.0 μM, and the inhibition data best fitted a mixed-type mode with α = 0.6. The same extent of inhibition was observed at the cellular level in the human colorectal carcinoma HCT 116 cell line transfected with a plasmid encoding the AKR1C3 transcript (IC50 = 7.9 μM). Furthermore, we performed an analysis of flexible docking between buparlisib and AKR1C3 and found that buparlisib probably occupies a part of the binding site for a cofactor most likely via the trifluoromethyl group of buparlisib interacting with catalytic residue Tyr55. In conclusion, our results show a novel PI3K-independent effect of buparlisib that may improve therapeutic efficacy and safety of daunorubicin by preventing its metabolism by AKR1C3.

• Klíčová slova
• AKR1C3, Anthracyclines, Buparlisib, Cancer, HCT116 cell line, Multidrug resistance,
• MeSH
• aldo-keto reduktasy antagonisté a inhibitory   genetika   metabolismus   MeSH
• aminopyridiny chemie   metabolismus   farmakologie   MeSH
• daunomycin metabolismus   MeSH
• HCT116 buňky MeSH
• inhibiční koncentrace 50 MeSH
• katalytická doména MeSH
• kinetika MeSH
• lidé MeSH
• morfoliny chemie   metabolismus   farmakologie   MeSH
• protein AKR1C3 antagonisté a inhibitory   chemie   metabolismus   MeSH
• rekombinantní proteiny biosyntéza   izolace a purifikace   metabolismus   MeSH
• simulace molekulového dockingu MeSH
• vazebná místa MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aldo-keto reduktasy MeSH
• aminopyridiny MeSH
• daunomycin MeSH
• morfoliny MeSH
• NVP-BKM120 MeSH Prohlížeč
• protein AKR1C3 MeSH
• rekombinantní proteiny MeSH