During development, tooth germs undergo various morphological changes resulting from interactions between the oral epithelium and ectomesenchyme. These processes are influenced by the extracellular matrix, the composition of which, along with cell adhesion and signaling, is regulated by metalloproteinases. Notably, these include matrix metalloproteinases (MMPs), a disintegrin and metalloproteinases (ADAMs), and a disintegrin and metalloproteinases with thrombospondin motifs (ADAMTSs). Our analysis of previously published scRNAseq datasets highlight that these metalloproteinases show dynamic expression patterns during tooth development, with expression in a wide range of cell types, suggesting multiple roles in tooth morphogenesis. To investigate this, Marimastat, a broad-spectrum inhibitor of MMPs, ADAMs, and ADAMTSs, was applied to ex vivo cultures of mouse molar tooth germs. The treated samples exhibited significant changes in tooth germ size and morphology, including an overall reduction in size and an inversion of the typical bell shape. The cervical loop failed to extend, and the central area of the inner enamel epithelium protruded. Marimastat treatment also disrupted proliferation, cell polarization, and organization compared with control tooth germs. In addition, a decrease in laminin expression was observed, leading to a disruption in continuity of the basement membrane at the epithelial-mesenchymal junction. Elevated hypoxia-inducible factor 1-alpha gene (Hif-1α) expression correlated with a disruption to blood vessel development around the tooth germs. These results reveal the crucial role of metalloproteinases in tooth growth, shape, cervical loop elongation, and the regulation of blood vessel formation during prenatal tooth development.NEW & NOTEWORTHY Inhibition of metalloproteinases during tooth development had a wide-ranging impact on molar growth affecting proliferation, cell migration, and vascularization, highlighting the diverse role of these proteins in controlling development.

• Klíčová slova
• blood vessels, cervical loop, extracellular matrix, metalloproteinases, tooth germ morphogenesis,
• MeSH
• faktor 1 indukovatelný hypoxií - podjednotka alfa metabolismus   genetika   MeSH
• inhibitory matrixových metaloproteinas farmakologie   MeSH
• kyseliny hydroxamové farmakologie   MeSH
• metaloproteasy metabolismus   genetika   MeSH
• moláry embryologie   růst a vývoj   metabolismus   enzymologie   MeSH
• morfogeneze MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• odontogeneze * MeSH
• proliferace buněk * MeSH
• vývojová regulace genové exprese MeSH
• zubní zárodek embryologie   metabolismus   enzymologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• faktor 1 indukovatelný hypoxií - podjednotka alfa MeSH
• inhibitory matrixových metaloproteinas MeSH
• kyseliny hydroxamové MeSH
• marimastat MeSH Prohlížeč
• metaloproteasy MeSH

The pathogenic fungus Aspergillus fumigatus utilizes a cyclic ferrioxamine E (FOXE) siderophore to acquire iron from the host. Biomimetic FOXE analogues were labeled with gallium-68 for molecular imaging with PET. [68Ga]Ga(III)-FOXE analogues were internalized in A. fumigatus cells via Sit1. Uptake of [68Ga]Ga(III)-FOX 2-5, the most structurally alike analogue to FOXE, was high by both A. fumigatus and bacterial Staphylococcus aureus. However, altering the ring size provoked species-specific uptake between these two microbes: ring size shortening by one methylene unit (FOX 2-4) increased uptake by A. fumigatus compared to that by S. aureus, whereas lengthening the ring (FOX 2-6 and 3-5) had the opposite effect. These results were consistent both in vitro and in vivo, including PET imaging in infection models. Overall, this study provided valuable structural insights into the specificity of siderophore uptake and, for the first time, opened up ways for selective targeting and imaging of microbial pathogens by siderophore derivatization.

• MeSH
• Aspergillus fumigatus * metabolismus   chemie   MeSH
• aspergilóza * diagnostické zobrazování   mikrobiologie   MeSH
• biomimetické materiály chemie   metabolismus   MeSH
• cyklické peptidy MeSH
• deferoxamin chemie   MeSH
• druhová specificita MeSH
• myši MeSH
• pozitronová emisní tomografie * metody   MeSH
• radioizotopy galia * chemie   MeSH
• siderofory * chemie   metabolismus   MeSH
• Staphylococcus aureus * metabolismus   MeSH
• železité sloučeniny chemie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cyklické peptidy MeSH
• deferoxamin MeSH
• ferrioxamine E MeSH Prohlížeč
• Gallium-68 MeSH Prohlížeč
• radioizotopy galia * MeSH
• siderofory * MeSH
• železité sloučeniny MeSH

Because of synergism between tubulin and HDAC inhibitors, we used the pharmacophore fusion strategy to generate potential tubulin-HDAC dual inhibitors. Drug design was based on the introduction of a N-hydroxyacrylamide or a N-hydroxypropiolamide at the 5-position of the 2-aroylbenzo[b]furan skeleton, to produce compounds 6a-i and 11a-h, respectively. Among the synthesized compounds, derivatives 6a, 6c, 6e, 6g, 11a, and 11c showed excellent antiproliferative activity, with IC50 values at single- or double-digit nanomolar levels, against the A549, HT-29, and MCF-7 cells resistant towards the control compound combretastatin A-4 (CA-4). Compounds 11a and 6g were also 10-fold more active than CA-4 against the Hela cell line. When comparing the inhibition of tubulin polymerization versus the HDAC6 inhibitory activity, we found that 6a-g, 6i, 11a, 11c, and 11e, although very potent as inhibitors of tubulin assembly, did not have significant inhibitory activity against HDAC6.

• Klíčová slova
• anticancer agents, benzo[b]furan, dual-target inhibitors, histone deacetylase (HDAC), tubulin,
• MeSH
• antitumorózní látky * farmakologie   chemická syntéza   chemie   MeSH
• benzofurany * farmakologie   chemie   chemická syntéza   MeSH
• buňky HT-29 MeSH
• HeLa buňky MeSH
• histondeacetylasa 6 antagonisté a inhibitory   metabolismus   MeSH
• inhibitory histondeacetylas farmakologie   chemická syntéza   chemie   MeSH
• kyseliny hydroxamové * farmakologie   chemie   chemická syntéza   MeSH
• léky antitumorózní - screeningové testy MeSH
• lidé MeSH
• MFC-7 buňky MeSH
• modulátory tubulinu * farmakologie   chemická syntéza   chemie   MeSH
• nádorové buněčné linie MeSH
• proliferace buněk * účinky léků   MeSH
• tubulin * metabolismus   MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antitumorózní látky * MeSH
• benzofuran MeSH Prohlížeč
• benzofurany * MeSH
• histondeacetylasa 6 MeSH
• inhibitory histondeacetylas MeSH
• kyseliny hydroxamové * MeSH
• modulátory tubulinu * MeSH
• tubulin * MeSH

Iron, as an essential micronutrient, plays a crucial role in host-pathogen interactions. In order to limit the growth of the pathogen, a common strategy of innate immunity includes withdrawing available iron to interfere with the cellular processes of the microorganism. Against that, unicellular parasites have developed powerful strategies to scavenge iron, despite the effort of the host. Iron-sequestering compounds, such as the approved and potent chelator deferoxamine (DFO), are considered a viable option for therapeutic intervention. Since iron is heavily utilized in the mitochondrion, targeting iron chelators in this organelle could constitute an effective therapeutic strategy. This work presents mitochondrially targeted DFO, mitoDFO, as a candidate against a range of unicellular parasites with promising in vitro efficiency. Intracellular Leishmania infection can be cleared by this compound, and experimentation with Trypanosoma brucei 427 elucidates its possible mode of action. The compound not only affects iron homeostasis but also alters the physiochemical properties of the inner mitochondrial membrane, resulting in a loss of function. Furthermore, investigating the virulence factors of pathogenic yeasts confirms that mitoDFO is a viable candidate for therapeutic intervention against a wide spectrum of microbe-associated diseases.

• Klíčová slova
• chelation, iron, mitochondria, parasites, protists,
• MeSH
• antiinfekční látky * MeSH
• antiparazitární látky farmakologie   MeSH
• chelátory železa farmakologie   terapeutické užití   MeSH
• deferoxamin chemie   MeSH
• mitochondrie MeSH
• železo * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• antiinfekční látky * MeSH
• antiparazitární látky MeSH
• chelátory železa MeSH
• deferoxamin MeSH
• železo * MeSH

Histone deacetylase (HDAC) inhibitors used in the clinic typically contain a hydroxamate zinc-binding group (ZBG). However, more recent work has shown that the use of alternative ZBGs, and, in particular, the heterocyclic oxadiazoles, can confer higher isoenzyme selectivity and more favorable ADMET profiles. Herein, we report on the synthesis and biochemical, crystallographic, and computational characterization of a series of oxadiazole-based inhibitors selectively targeting the HDAC6 isoform. Surprisingly, but in line with a very recent finding reported in the literature, a crystal structure of the HDAC6/inhibitor complex revealed that hydrolysis of the oxadiazole ring transforms the parent oxadiazole into an acylhydrazide through a sequence of two hydrolytic steps. An identical cleavage pattern was also observed both in vitro using the purified HDAC6 enzyme as well as in cellular systems. By employing advanced quantum and molecular mechanics (QM/MM) and QM calculations, we elucidated the mechanistic details of the two hydrolytic steps to obtain a comprehensive mechanistic view of the double hydrolysis of the oxadiazole ring. This was achieved by fully characterizing the reaction coordinate, including identification of the structures of all intermediates and transition states, together with calculations of their respective activation (free) energies. In addition, we ruled out several (intuitively) competing pathways. The computed data (ΔG‡ ≈ 21 kcal·mol-1 for the rate-determining step of the overall dual hydrolysis) are in very good agreement with the experimentally determined rate constants, which a posteriori supports the proposed reaction mechanism. We also clearly (and quantitatively) explain the role of the -CF3 or -CHF2 substituent on the oxadiazole ring, which is a prerequisite for hydrolysis to occur. Overall, our data provide compelling evidence that the oxadiazole warheads can be efficiently transformed within the active sites of target metallohydrolases to afford reaction products possessing distinct selectivity and inhibition profiles.

• MeSH
• histondeacetylasa 6 chemie   MeSH
• hydrolýza MeSH
• inhibitory histondeacetylas * farmakologie   MeSH
• kyseliny hydroxamové chemie   MeSH
• oxadiazoly * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• histondeacetylasa 6 MeSH
• inhibitory histondeacetylas * MeSH
• kyseliny hydroxamové MeSH
• oxadiazoly * MeSH

Histone deacetylase 6 (HDAC6) is a unique member of the HDAC family of enzymes due to its complex domain organization and cytosolic localization. Experimental data point toward the therapeutic use of HDAC6-selective inhibitors (HDAC6is) for use in both neurological and psychiatric disorders. In this article, we provide side-by-side comparisons of hydroxamate-based HDAC6is frequently used in the field and a novel HDAC6 inhibitor containing the difluoromethyl-1,3,4-oxadiazole function as an alternative zinc-binding group (compound 7). In vitro isotype selectivity screening uncovered HDAC10 as a primary off-target for the hydroxamate-based HDAC6is, while compound 7 features exquisite 10,000-fold selectivity over all other HDAC isoforms. Complementary cell-based assays using tubulin acetylation as a surrogate readout revealed approximately 100-fold lower apparent potency for all compounds. Finally, the limited selectivity of a number of these HDAC6is is shown to be linked to cytotoxicity in RPMI-8226 cells. Our results clearly show that off-target effects of HDAC6is must be considered before attributing observed physiological readouts solely to HDAC6 inhibition. Moreover, given their unparalleled specificity, the oxadiazole-based inhibitors would best be employed either as research tools in further probing HDAC6 biology or as leads in the development of truly HDAC6-specific compounds in the treatment of human disease states.

• Klíčová slova
• histone deacetylase, inhibitor profiling, metallohydrolase, nanoBRET, tubulin/histone acetylation,
• MeSH
• acetylace MeSH
• histondeacetylasa 6 * antagonisté a inhibitory   MeSH
• histondeacetylasy * metabolismus   MeSH
• inhibitory histondeacetylas * chemie   farmakologie   MeSH
• kyseliny hydroxamové * chemie   farmakologie   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• oxadiazoly * chemie   farmakologie   MeSH
• posttranslační úpravy proteinů MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• HDAC10 protein, human MeSH Prohlížeč
• histondeacetylasa 6 * MeSH
• histondeacetylasy * MeSH
• inhibitory histondeacetylas * MeSH
• kyseliny hydroxamové * MeSH
• oxadiazoly * MeSH

Mycobacterial zinc metalloprotease-1 (Zmp1) is an essential enzyme for intracellular survival and pathogenicity of Mycobacterium tuberculosis. However, the exact mechanism of function of this enzyme remains unclear. This paper examines the effect of novel organic molecules on the inhibition of Zmp1. We followed our previous results and synthesised three libraries of new hydroxamates. All compounds were studied for their inhibitory properties towards a recombinant Zmp1 from Mycobacterium tuberculosis by MALDI-TOF MS. Furthermore, a macrophage infection assay was performed to evaluate intracellular antimycobacterial activity. In the whole-cell assay, no direct activity of synthesised heterocyclic hydroxamates was observed against Mycobacterium tuberculosis and Mycobacterium bovis. No acute cellular toxicity was observed against the murine RAW 264.7 macrophage cell line and human MRC-5 lung fibroblast cell line. However, thiazolidinediones 2 showed the dose-dependent inhibition of intracellular survival of Mycobacterium tuberculosis H37Ra. The inhibition was structure-dependent, with the most active derivative 2f inducing an 83.2% reduction of bacterial survival within the macrophage host cell. The promising biological activity confirmed thiazolidinediones 2 as Zmp1 inhibitors that can be used as tool compounds for further exploration of the role of Zmp1 for in vivo pathogenicity. In the long run, thiazolidinediones 2 show the potential to act as a scaffold for Zmp1 inhibitors to target intracellular Mtb as a novel tuberculosis treatment strategy.

• Klíčová slova
• Hydroxamate, Indole, Mycobacterium tuberculosis, Pyrrole, Thiazolidinedione, Virulence factor Zmp1,
• MeSH
• bakteriální proteiny MeSH
• kyseliny hydroxamové farmakologie   MeSH
• lidé MeSH
• metaloproteasy metabolismus   MeSH
• Mycobacterium tuberculosis * MeSH
• myši MeSH
• thiazolidindiony * farmakologie   MeSH
• zinek metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• bakteriální proteiny MeSH
• kyseliny hydroxamové MeSH
• metaloproteasy MeSH
• thiazolidindiony * MeSH
• zinek MeSH

Dysregulation of iron homeostasis is one of the important processes in the development of many oncological diseases, such as pancreatic cancer. Targeting it with specific agents, such as an iron chelator, are promising therapeutic methods. In this study, we tested the cytotoxicity of novel azulene hydrazide-hydrazone-based chelators against pancreatic cancer cell lines (MIA PaCa-2, PANC-1, AsPC-1). All prepared chelators (compounds 4-6) showed strong cytotoxicity against pancreatic cancer cell lines and high selectivity for cancer cell lines compared to the healthy line. Their cytotoxicity is lower than thiosemicarbazone-based chelators Dp44mT and DpC, but significantly higher than hydroxamic acid-based chelator DFO. The chelator tested showed mitochondrial and lysosomal co-localization and its mechanism of action was based on the induction of hypoxia-inducible factor-1-alpha (HIF-1α), N-myc downstream-regulated gene-1 (NDRG1) and transferrin receptor 1 (TfR1). This strongly implies that the cytotoxic effect of tested chelators could be associated with mitophagy induction. Lipinski's rule of five analyses was performed to determine whether the prepared compounds had properties ensuring their bioavailability. In addition, the drug-likeness and drug-score were calculated and discussed.

• Klíčová slova
• Cancer, Chelators, HIF-1α, Hydrazone, NDRG1, TfR1,
• MeSH
• azuleny MeSH
• chelátory železa farmakologie   MeSH
• hydraziny MeSH
• hydrazony farmakologie   MeSH
• kyseliny hydroxamové MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory slinivky břišní * farmakoterapie   MeSH
• receptory transferinu MeSH
• thiosemikarbazony * farmakologie   MeSH
• železo MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• azulene MeSH Prohlížeč
• azuleny MeSH
• chelátory železa MeSH
• hydraziny MeSH
• hydrazony MeSH
• kyseliny hydroxamové MeSH
• receptory transferinu MeSH
• thiosemikarbazony * MeSH
• železo MeSH

Histone deacetylases (HDACs) target acetylated lysine residues in histone and non-histone proteins. HDACs are implicated in the regulation of genomic stability, cell cycle, cell death and differentiation and thus critically involved in tumorigenesis. Further, HDACs regulate T-cell development and HDAC inhibitors (HDACis) have been approved for clinical use in some T-cell malignancies. Still, the exact targets and mechanisms of HDAC inhibition in cancer are understudied. We isolated tumor cell lines from a transgenic mouse model of anaplastic large cell lymphoma (ALCL), a rare T-cell lymphoma, and abrogated HDAC activity by treatment with the HDACis Vorinostat and Entinostat or Cre-mediated deletion of Hdac1. Changes in overall protein expression as well as histone and protein acetylation were measured following Hdac1 deletion or pharmacological inhibition using label-free liquid chromatography mass spectrometry (LC-MS/MS). We found changes in overall protein abundance and increased acetylation of histones and non-histone proteins, many of which were newly discovered and associated with major metabolic and DNA damage pathways. For non-histone acetylation, we mapped a total of 1204 acetylated peptides corresponding to 603 proteins, including chromatin modifying proteins and transcription factors. Hyperacetylated proteins were involved in processes such as transcription, RNA metabolism and DNA damage repair (DDR). The DDR pathway was majorly affected by hyperacetylation following HDAC inhibition. This included acetylation of H2AX, PARP1 and previously unrecognized acetylation sites in TP53BP1. Our data provide a comprehensive view of the targets of HDAC inhibition in malignant T cells with general applicability and could have translational impact for the treatment of ALCL with HDACis alone or in combination therapies.

• Klíčová slova
• ALCL, MS-275, SAHA, acetylomics, anaplastic large cell lymphoma, entinostat, histone deacetylase inhibitors, histone deacetylases, proteomics, vorinostat,
• MeSH
• acetylace MeSH
• anaplastický velkobuněčný lymfom * farmakoterapie   MeSH
• chromatografie kapalinová MeSH
• histondeacetylasy * metabolismus   MeSH
• histony metabolismus   MeSH
• kyseliny hydroxamové farmakologie   MeSH
• myši MeSH
• tandemová hmotnostní spektrometrie MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• histondeacetylasy * MeSH
• histony MeSH
• kyseliny hydroxamové MeSH

• Klíčová slova
• Actinonin, Matrix metalloproteinases, Metastases, Polymer conjugate, pH-sensitive release,
• MeSH
• doxorubicin farmakologie   MeSH
• kyseliny hydroxamové MeSH
• lidé MeSH
• methakryláty MeSH
• nádory * farmakoterapie   MeSH
• nanomedicína * MeSH
• polymery farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• actinonin MeSH Prohlížeč
• doxorubicin MeSH
• hydroxypropyl methacrylate MeSH Prohlížeč
• kyseliny hydroxamové MeSH
• methakryláty MeSH
• polymery MeSH