Interleukin-2-inducible T-cell kinase (ITK) and Bruton's tyrosine kinase (BTK) are two important members of the Tec family with crucial roles in immune system function. Deregulation in ITK and BTK activity is linked to several hematological malignancies, making them key targets for cancer immunotherapy. In this study, we synthesized a series of azaspirooxindolinone derivatives and evaluated their cytotoxic activity against ITK/BTK-negative and positive cancer cell lines, followed by enzymatic inhibition studies to assess the ITK/BTK kinase selectivity of two hit compounds. Several compounds demonstrated selective cytotoxicity against ITK- or BTK-expressing cells. Compound 3d exhibited high cytotoxicity in ITK-positive Jurkat (IC50 = 3.58 µM) and BTK-positive Ramos (IC50 = 3.06 µM) cells, while compound 3j showed strong cytotoxicity in Ramos (IC50 = 1.38 µM) and Jurkat (IC50 = 4.16 µM) cells. Compounds 3a and 3e were selectively cytotoxic in Jurkat cells (IC50 = 9.36 µM and 10.85 µM, respectively), while compounds 3f and 3g were highly cytotoxic in Ramos cells (IC50 = 1.82 µM and 1.42 µM, respectively). None of the active compounds exhibited cytotoxicity in non-cancer cell lines (IC50 > 50 µM), demonstrating their selectivity for malignant cells. Enzyme inhibition assay showed that 3d is a selective ITK inhibitor (IC50 = 0.91 µM) with no detectable BTK inhibition, aligning with its strong activity in ITK-positive cells. In contrast, compound 3j did not inhibit ITK or BTK enzymatically, suggesting an alternative mechanism of action. These findings highlight 3d as a promising ITK inhibitor and warrant further investigation to elucidate its mechanism of action.

• Klíčová slova
• Anti-cancer derivatives, Azaspirooxindolinones, Bruton’s tyrosine kinase, Interleukin-2-inducible T-cell kinase, Molecular docking,
• MeSH
• inhibitory proteinkinas * farmakologie   chemická syntéza   chemie   MeSH
• lidé MeSH
• molekulární struktura MeSH
• nádorové buněčné linie MeSH
• oxindoly farmakologie   chemie   chemická syntéza   MeSH
• proliferace buněk účinky léků   MeSH
• proteinkinasa BTK * antagonisté a inhibitory   metabolismus   MeSH
• protinádorové látky * farmakologie   chemická syntéza   chemie   MeSH
• racionální návrh léčiv * MeSH
• screeningové testy protinádorových léčiv * MeSH
• simulace molekulového dockingu MeSH
• spirosloučeniny chemie   farmakologie   chemická syntéza   MeSH
• tyrosinkinasy * antagonisté a inhibitory   metabolismus   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
• BTK protein, human MeSH Prohlížeč
• emt protein-tyrosine kinase MeSH Prohlížeč
• inhibitory proteinkinas * MeSH
• oxindoly MeSH
• proteinkinasa BTK * MeSH
• protinádorové látky * MeSH
• spirosloučeniny MeSH
• tyrosinkinasy * MeSH

In the complex network of cellular physiology, the maintenance of cellular proteostasis emerges as a critical factor for cell survival, particularly under stress conditions. This homeostasis is largely governed by a sophisticated network of molecular chaperones and co-chaperones, among which Bcl-2-associated athanogene 3 (BAG3), able to interact with the ATPase domain of Heat Shock Protein 70 (HSP70), plays a pivotal role. The BAG3-HSP70 functional module is not only essential for cellular homeostasis but is also involved in the pathogenesis of various diseases, including cancer, neurodegenerative disorders, and cardiac dysfunction, making it an attractive target for therapeutic intervention. Inspired by our continuous interest in the development of new chemical platforms able to interfere with BAG3 protein, herein we report the discovery of compound 16, the first-in-class BAG3/HSP70 dual modulator, obtained by combining the multicomponent Ugi reaction with the alkyne-azide Huisgen procedure in a sequential tandem reaction approach. Through a combination of biophysical analysis, biochemical assays, and cell-based studies, we elucidated the mechanism of action of this inhibitor and assessed its potential as a therapeutic agent. Hence, this study can open new avenues for the development of novel anticancer strategies that leverage the simultaneous disruption of multiple chaperone pathways.

• Klíčová slova
• Anticancer agent, BAG3, Dual modulator, HSP70, Ugi-Huisgen tandem approach,
• MeSH
• adaptorové proteiny signální transdukční * metabolismus   antagonisté a inhibitory   MeSH
• lidé MeSH
• molekulární chaperony metabolismus   antagonisté a inhibitory   chemie   MeSH
• molekulární struktura MeSH
• nádorové buněčné linie MeSH
• proliferace buněk účinky léků   MeSH
• proteiny regulující apoptózu * metabolismus   antagonisté a inhibitory   MeSH
• proteiny tepelného šoku HSP70 * antagonisté a inhibitory   metabolismus   MeSH
• protinádorové látky * farmakologie   chemie   chemická syntéza   MeSH
• screeningové testy protinádorových 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
• adaptorové proteiny signální transdukční * MeSH
• BAG3 protein, human MeSH Prohlížeč
• molekulární chaperony MeSH
• proteiny regulující apoptózu * MeSH
• proteiny tepelného šoku HSP70 * MeSH
• protinádorové látky * MeSH

A new group of potent histone deacetylase inhibitors (HDACis) capable of inhibiting cell growth and affecting cell-cycle progression in Tohoku Hospital Pediatrics-1 (THP-1) monocytic leukaemia cells was synthesized. The inhibitors belong to a series of hydroxamic acid derivatives. We designed and synthesized a series of 22 N-hydroxycinnamamide derivatives, out of which 20 are new compounds. These compounds contain various substituted anilides as the surface recognition moiety (SRM), a p-hydroxycinnamate linker, and hydroxamic acids as the zinc-binding group (ZBG). The whole series of synthesized hydroxamic acids inhibited THP-1 cell proliferation. Compounds 7d and 7p, which belong to the category of derivatives with the most potent antiproliferative properties, exert a similar effect on cell-cycle progression as vorinostat and induce apoptosis in THP-1 cells. Furthermore, compounds 7d and 7p were demonstrated to inhibit HDAC class I and II in THP-1 cells with comparable potency to vorinostat and increase acetylation of histones H2a, H2b, H3, and H4. Molecular modelling was used to predict the probable binding mode of the studied HDACis in class I and II histone deacetylases in terms of Zn2+ ion chelation by the hydroxamate group.

• Klíčová slova
• HDACi, anticancer agents, haematological malignancies, hydroxamic acid, inhibitors of histone deacetylases,
• MeSH
• apoptóza * účinky léků   MeSH
• buněčný cyklus účinky léků   MeSH
• histondeacetylasy metabolismus   MeSH
• inhibitory histondeacetylas * farmakologie   chemická syntéza   chemie   MeSH
• kyseliny hydroxamové farmakologie   chemická syntéza   chemie   MeSH
• kyseliny kumarové farmakologie   chemická syntéza   chemie   MeSH
• lidé MeSH
• molekulární struktura MeSH
• proliferace buněk * účinky léků   MeSH
• protinádorové látky * farmakologie   chemická syntéza   chemie   MeSH
• screeningové testy protinádorových léčiv MeSH
• simulace molekulového dockingu MeSH
• THP-1 buňky MeSH
• vorinostat farmakologie   chemická syntéza   chemie   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
• histondeacetylasy MeSH
• inhibitory histondeacetylas * MeSH
• kyseliny hydroxamové MeSH
• kyseliny kumarové MeSH
• protinádorové látky * MeSH
• vorinostat MeSH

Anaplastic Large Cell Lymphoma (ALCL) is an aggressive T-cell lymphoma affecting children and young adults. About 30% of patients develop therapy resistance therefore new precision medicine drugs are highly warranted. Multiple rounds of structure-activity optimization of Caffeic Acid Phenethyl Ester have resulted in CM14. CM14 causes upregulation of genes involved in oxidative stress response and downregulation of DNA replication genes leading to G2/M arrest and subsequent apoptosis induction. In accordance with this, an unbiased proteomics approach, confocal microscopy and molecular modeling showed that TUBGCP2, member of the centrosomal γ-TuRC complex, is a direct interaction partner of CM14. CM14 overcomes ALK inhibitor resistance in ALCL and is also active in T-cell Acute Lymphoblastic Leukemia and Acute Myeloid Leukemia. Interestingly, CM14 also induced cell death in docetaxel-resistant prostate cancer cells thus suggesting an unexpected role in solid cancers. Thus, we synthesized and thoroughly characterized a novel TUBGCP2 targeting drug that is active in ALCL but has also potential for other malignancies.

• MeSH
• apoptóza * účinky léků   MeSH
• centrozom * účinky léků   metabolismus   MeSH
• fenethylalkohol * analogy a deriváty   farmakologie   chemie   MeSH
• kyseliny kávové * farmakologie   chemie   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• proliferace buněk účinky léků   MeSH
• protinádorové látky * farmakologie   chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• caffeic acid phenethyl ester MeSH Prohlížeč
• fenethylalkohol * MeSH
• kyseliny kávové * MeSH
• protinádorové látky * MeSH

Developing methodological approaches for discovering novel pathways is a key challenge in the life science research. Biological pathways are regulated-in higher eukaryotes-by a vast diversity of linear peptide motifs that mediate combinatorial specificity in signal transduction pathways. The E3 ubiquitin ligase component (MDM2) is such a protein that interacts with target proteins containing linear motifs such as p53. Drug leads, such as Nutlin-3, that bind to the MDM2 hydrophobic pocket mimic p53 and can release p53 from MDM2 control and this can lead to cell death. However, these drug leads act allosterically, having agonist effects on MDM2's functions and there are other proteins whose steady state levels can be altered by Nutlin-3. As cell density can alter the proliferation state of cell populations, we examined the impact of Nutlin-3 on levels of newly synthesized proteins using pulse-SILAC mass spectrometry. The data demonstrate that at differing cell densities or population-wide proliferation rates, different newly synthesized proteins dominate the proteome landscape in a Nutlin-3 dependent manner. These data further confirm that the cell state in a population of cells can in turn impact on the MDM2 signalling landscape. This methodology forms a blueprint for biomarker discovery using clinical samples that can detect changes in the synthesis rate of proteins in cell populations treated with specific agents. Broader implications highlight tools that can be used to study allosteric regulation of protein-drug combinations.

• Klíčová slova
• MDM2, P53, biomarker discovery, mass spectrometry, pulse‐SILAC,
• MeSH
• imidazoly * farmakologie   MeSH
• lidé MeSH
• nádorový supresorový protein p53 metabolismus   MeSH
• piperaziny * farmakologie   MeSH
• proliferace buněk účinky léků   MeSH
• proteom * metabolismus   MeSH
• proteomika metody   MeSH
• protoonkogenní proteiny c-mdm2 metabolismus   MeSH
• signální transdukce účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• imidazoly * MeSH
• MDM2 protein, human MeSH Prohlížeč
• nádorový supresorový protein p53 MeSH
• nutlin 3 MeSH Prohlížeč
• piperaziny * MeSH
• proteom * MeSH
• protoonkogenní proteiny c-mdm2 MeSH

Bruton's Tyrosine Kinase (BTK) is a key driver of hematological malignancies, autoimmune disorders, and neuroinflammation, making it an attractive therapeutic target. Proteolysis targeting chimeras (PROTACs) offer a novel strategy for BTK degradation via the E3 ubiquitin ligase pathway. Here, we evaluated nine azaspirooxindolinone-based PROTAC derivatives for their cytotoxicity and BTK-targeting activity. Several compounds exhibited potent cytotoxicity against BTK-high RAMOS lymphoma cells without affecting non-cancer fibroblasts or normal T/B-cell lymphocytes. Among them, PROTAC 25 emerged as the most effective degraded, achieving a Dmax of 72.84 % and DC50 of 0.27 μM in a proteasome-dependent manner. Although PROTAC 25 was cytotoxic to IL-2-inducible T cell Kinase (ITK)-positive cells, ITK protein levels remained unaffected. Furthermore, kinase assays revealed that PROTAC 25 inhibited BTK kinase activity (IC₅₀ = 0.44 μM) with moderate selectivity over ITK (IC₅₀ = 2.16 μM). Notably, PROTAC 25 suppressed BTK-mediated downstream signaling in RAMOS cells, as evidenced by reduced phosphorylation of BTK and its downstream effector, p38 MAPK. These findings highlight PROTAC 25 as a promising BTK degrader with therapeutic potential and underscore the value of azaspirooxindolinone-based PROTACs in targeting BTK-driven diseases.

• Klíčová slova
• Antiproliferative activity, Azaspirooxindolinone, Bruton's tyrosine kinase, IL-2-inducible T-cell kinase, Jurkat, PROTAC, Ramos,
• MeSH
• inhibitory proteinkinas farmakologie   chemie   chemická syntéza   MeSH
• lidé MeSH
• molekulární struktura MeSH
• nádorové buněčné linie MeSH
• oxindoly farmakologie   chemie   chemická syntéza   MeSH
• proliferace buněk účinky léků   MeSH
• proteinkinasa BTK * antagonisté a inhibitory   metabolismus   MeSH
• proteolýza * účinky léků   MeSH
• protinádorové látky * farmakologie   chemie   chemická syntéza   MeSH
• screeningové testy protinádorových 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
• BTK protein, human MeSH Prohlížeč
• inhibitory proteinkinas MeSH
• oxindoly MeSH
• proteinkinasa BTK * MeSH
• protinádorové látky * MeSH

For the treatment of bilateral limbal stem cell deficiency (LSCD), cell therapy with transplantation of cultivated oral mucosa epithelial cells (COMET) is a promising alternative. Although not yet established, current protocols on the cultivation of oral mucosal epithelial cell (OMECs) sheets are based mainly on substrates and xenobiotic additives that may lead to variable outcomes and undesirable immune responses by the patient. The aim of this study was to characterize OMECs cultivated in xenobiotic-free media (XF) seeded on fibrin gel, in comparison to conventional complex (COM) medium. Oral mucosal biopsies were retrieved from 31 donors. After cultivation in COM or XF medium, OMECs were compared based on growth kinetics, morphology, cell size and viability. Using immunofluorescence and gene expression analyses, the degree of stemness, proliferation and differentiation was evaluated in OMEC cultures. Our findings showed that although OMECs showed a similar morphology and viability, and comparable growth kinetics, immunofluorescence revealed the preservation of stemness (p63 + p40 positivity in cells ≤11 μm) and proliferation in both COM and XF. Gene expression analyses showed that keratin (K)13 and K15 expression levels were significantly higher in XF (adj. p < 0.001), but otherwise COM and XF-treated OMECs had comparable transcriptional profiles in a panel of stemness, proliferation and differentiation genes. These results demonstrate the feasibility of culturing OMECs on fibrin gel without xenogeneic additives, while maintaining their undifferentiated state and preserving stemness. In conclusion, both in terms of results and methodology, the procedures presented here are suitable for implementation in clinical practice.

• Klíčová slova
• Advanced therapy medicinal products, Cell culture, Fibrin glue substrate, Limbal stem cell deficiency, Oral mucosal epithelial cells, Stemness,
• MeSH
• buněčná diferenciace MeSH
• buněčné kultury * MeSH
• deficit limbálních kmenových buněk MeSH
• dospělí MeSH
• epitelové buňky * metabolismus   účinky léků   MeSH
• fibrin * MeSH
• gely MeSH
• kmenové buňky * metabolismus   cytologie   MeSH
• kultivační média MeSH
• kultivované buňky MeSH
• lidé středního věku MeSH
• lidé MeSH
• limbus corneae * cytologie   patologie   metabolismus   MeSH
• nemoci rohovky patologie   farmakoterapie   metabolismus   MeSH
• proliferace buněk účinky léků   MeSH
• rohovkový epitel metabolismus   cytologie   účinky léků   patologie   MeSH
• senioři MeSH
• transplantace kmenových buněk metody   MeSH
• ústní sliznice * cytologie   MeSH
• viabilita buněk MeSH
• xenobiotika farmakologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fibrin * MeSH
• gely MeSH
• kultivační média MeSH
• xenobiotika MeSH

Development of electrospun nanofibers with suitable properties to promote wound healing is an advantage in developing non-invasive skin treatments. We showed the potential application of Polyvinyl acetate (PVA) and Polyvinylpyrrolidone (PVP) combined with Helichrysum italicum oil (HO) in wound healing. During this process, Tight junctions (TJs) play a crucial role in maintaining skin integrity. TJs are intercellular junctions composed of a variety of transmembrane proteins, including Occludin (OCLN), observed also in migrating epithelial cells. Changes in OCLN expression affect epidermal permeability, indicating an active role in the healing process. Within this context, we studied the OCLN expression during healing after scratch assay on Keratinocytes (HaCaT), by a confocal microscopic analysis. In addition, we evaluated the effect of treatment after scratch on cell elasticity by Atomic Force Microscopy (AFM) analysis. All results show a positive trend in cell proliferation and viability on HaCaT treated with functionalized nanofibers. These results were confirmed by the expression of genes involved in the early stages of the regenerative process. Understanding the cell mechanisms involved in skin changes during repair process would allow future application of nanomaterials combined with HO in vivo.

• Klíčová slova
• Bioactive molecules, Cellular mechanism, Keratinocytes, Nanofibers, Regenerative medicine, Tissue regeneration, Wound healing,
• MeSH
• buněčné linie MeSH
• hojení ran účinky léků   MeSH
• keratinocyty * účinky léků   metabolismus   cytologie   MeSH
• kůže účinky léků   metabolismus   MeSH
• lidé MeSH
• mikroskopie atomárních sil MeSH
• nanovlákna * chemie   MeSH
• okludin metabolismus   genetika   MeSH
• oleje rostlin farmakologie   chemie   MeSH
• polyvinylalkohol chemie   MeSH
• povidon * chemie   farmakologie   MeSH
• proliferace buněk * účinky léků   MeSH
• pružnost účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• okludin MeSH
• oleje rostlin MeSH
• polyvinylalkohol MeSH
• povidon * MeSH

Nowadays, various forms of organosilane materials are well established in the field of regenerative medicine, but interestingly, fibrous organosilanes have yet to be described. So far, technological obstacles prevent the preparation of such fibrous materials without any presence of spinnability-supporting organic polymers, various types of surfactants, or non-polar organic solvents, which are in many cases highly toxic and economically inconvenient. Recently, these obstacles were overcome by a complex, yet simple, technology combining different science perspectives from supramolecular chemistry through material science to tissue engineering. This paper suggests a synthesis of two biomedically promising monomeric organosilane precursors, N,N´-bis(3-(triethoxysilyl)propyl)terephthalamide (BTT) and N,N´-bis(3-(triethoxysilyl)propyl)pyridine-2,6-dicarboxamide (BTP), which are submitted to a sol-gel process combined with subsequent electrospinning technology. Such a unique procedure not only allows the preparation of toxic-free organosilane fibrous mats by suitable adjustment of sol-gel and electrospinning parameters but also simplifies material production via a one-pot synthesis approach further tuneable with appropriate organosilane precursors. The BTT and BTP fibrous materials prepared displayed not only a promising interface among the materials and 3T3 fibroblast cell lines but moreover, the interaction of nanofibrous materials with stem cells has yielded encouraging outcomes. Stem cell adhesion, proliferation, and differentiation were notably enhanced in the presence of these materials, suggesting a supportive microenvironment conducive to regenerative responses. The ability of the material to modulate the cellular behaviour of stem cells holds promising implications for the development of targeted and effective regenerative therapies.

• MeSH
• biokompatibilní materiály * chemie   MeSH
• buněčná adheze MeSH
• buněčná diferenciace účinky léků   MeSH
• lidé MeSH
• myši MeSH
• nanovlákna chemie   MeSH
• polymery chemie   MeSH
• proliferace buněk účinky léků   MeSH
• regenerace MeSH
• regenerativní lékařství * metody   MeSH
• silany chemie   MeSH
• testování materiálů MeSH
• tkáňové inženýrství * metody   MeSH
• tkáňové podpůrné struktury * chemie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biokompatibilní materiály * MeSH
• polymery MeSH
• silany MeSH

The novel diiron amine complexes [Fe2Cp2(CO)(NH2R')(μ-CO){μ-CN(Me)(Cy)}]CF3SO3 [R' = H, 3; Cy, 4; CH2CH2NH2, 5; CH2CH2NMe2, 6; CH2CH2(4-C6H4OMe), 7; CH2CH2(4-C6H4OH), 8; Cp = η5-C5H5, Cy = C6H11 = cyclohexyl] were synthesized in 49-92 % yields from [Fe2Cp2(CO)2(μ-CO){μ-CN(Me)(Cy)}]CF3SO3, 1a, using a straightforward two-step procedure. They were characterized by IR and multinuclear NMR spectroscopy, and the structure of 7 was confirmed through X-ray diffraction analysis. Complexes 3-8 and the acetonitrile adducts [Fe2Cp2(CO)(NCMe)(μ-CO){μ-CN(Me)(R)}]CF3SO3 (R = Cy, 2a; Me, 2b; Xyl = 2,6-C6H3Me2, 2c) were assessed for their water solubility, octanol-water partition coefficient and stability in physiological-like solutions. The in vitro antiproliferative activity of 2a-c and 3-8 was tested on seven human cancer cell lines (A2780, A2780R, PC3, A549, MCF7, HOS and HT-29), while the selectivity was evaluated using normal MRC-5 cells. Overall, the complexes exhibited variable cytotoxicity, with IC50 values reaching the low micromolar range for 3, 7 and 8 in A2780 and A2780R cells, along with significant selectivity. Targeted experiments covered cell cycle modification, induction of cell death, mitochondrial membrane potential, ROS production and interaction with DNA and bovine serum albumin (BSA) as a model protein. The interaction of 3 with BSA was further investigated through computational studies. Results showed a negligible increase in intracellular ROS levels (except for 2b) and insignificant changes in mitochondrial membrane potential.

• Klíčová slova
• Bioorganometallic chemistry, Cellular effects, Diiron complexes, In vitro cytotoxicity, Labile ligand,
• MeSH
• aminy chemie   farmakologie   MeSH
• komplexní sloučeniny chemie   farmakologie   chemická syntéza   MeSH
• lidé MeSH
• ligandy MeSH
• molekulární struktura MeSH
• nádorové buněčné linie MeSH
• proliferace buněk * účinky léků   MeSH
• protinádorové látky * farmakologie   chemie   chemická syntéza   MeSH
• screeningové testy protinádorových léčiv * MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• vztahy mezi strukturou a aktivitou MeSH
• železo chemie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aminy MeSH
• komplexní sloučeniny MeSH
• ligandy MeSH
• protinádorové látky * MeSH
• železo MeSH