Production of small RNAs by ribonuclease III Dicer is a key step in microRNA and RNA interference pathways, which employ Dicer-produced small RNAs as sequence-specific silencing guides. Further studies and manipulations of microRNA and RNA interference pathways would benefit from identification of small-molecule modulators. Here, we report a study of a fluorescence-based in vitro Dicer cleavage assay, which was adapted for high-throughput screening. The kinetic assay can be performed under single-turnover conditions (35 nM substrate and 70 nM Dicer) in a small volume (5 µL), which makes it suitable for high-throughput screening in a 1536-well format. As a proof of principle, a small library of bioactive compounds was analyzed, demonstrating potential of the assay.

• Klíčová slova
• Dicer, high-throughput screening, siRNA,
• MeSH
• buněčné linie MeSH
• fluorescenční spektrometrie metody   MeSH
• knihovny malých molekul MeSH
• lidé MeSH
• objevování léků MeSH
• reprodukovatelnost výsledků MeSH
• ribonukleasa III antagonisté a inhibitory   genetika   metabolismus   MeSH
• rychlé screeningové testy * MeSH
• substrátová specifita MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• knihovny malých molekul MeSH
• ribonukleasa III MeSH

Ebola hemorrhagic fever is a deadly disease caused by infection with one of the Ebola virus species. Although a significant progress has recently been made in understanding of Ebola virus biology and pathogenesis, development of effective anti-Ebola treatments has not been very productive, compared to other areas of antiviral research (e.g., HIV and HCV infections). No approved vaccine or medicine is available for Ebola but several are currently under development. This review summarises attempts in identification, evaluation, and development of small-molecule candidates for treatment of Ebola viral disease, including the most promising experimental drugs brincidofovir (CMX001), BCX4430, and favipiravir (T-705).

• Klíčová slova
• Ebola virus, Marburg virus, antiviral, filovirus, hemorrhagic fever,
• MeSH
• antivirové látky chemická syntéza   terapeutické užití   MeSH
• epidemický výskyt choroby MeSH
• hemoragická horečka Ebola farmakoterapie   epidemiologie   MeSH
• lidé MeSH
• rychlé screeningové testy MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Geografické názvy
• západní Afrika epidemiologie   MeSH
• Názvy látek
• antivirové látky MeSH

By binding to the spliceosomal protein Snu66, the human ubiquitin-like protein Hub1 is a modulator of the spliceosome performance and facilitates alternative splicing. Small molecules that bind to Hub1 would be of interest to study the protein-protein interaction of Hub1/Snu66, which is linked to several human pathologies, such as hypercholesterolemia, premature aging, neurodegenerative diseases, and cancer. To identify small molecule ligands for Hub1, we used the interface analysis, peptide modeling of the Hub1/Snu66 interaction and the fragment-based NMR screening. Fragment-based NMR screening has not proven sufficient to unambiguously search for fragments that bind to the Hub1 protein. This was because the Snu66 binding pocket of Hub1 is occupied by pH-sensitive residues, making it difficult to distinguish between pH-induced NMR shifts and actual binding events. The NMR analyses were therefore verified experimentally by microscale thermophoresis and by NMR pH titration experiments. Our study found two small peptides that showed binding to Hub1. These peptides are the first small-molecule ligands reported to interact with the Hub1 protein.

• Klíčová slova
• anti-cancer therapy, nuclear magnetic resonance, protein-peptide docking, protein-protein interactions, small-molecule inhibitors,
• MeSH
• alternativní sestřih * MeSH
• lidé MeSH
• ligandy MeSH
• magnetická rezonanční spektroskopie MeSH
• počítače MeSH
• spliceozomy * metabolismus   MeSH
• ubikvitiny genetika   MeSH
• vazba proteinů MeSH
• vazebná místa MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ligandy MeSH
• ubikvitiny MeSH

Aspartate transcarbamoylase (ATC) is the first committed step in de novo pyrimidine biosynthesis in eukaryotes and plants. A potent transition state analog of human ATCase (PALA) has previously been assessed in clinical trials for the treatment of cancer, but was ultimately unsuccessful. Additionally, inhibition of this pathway has been proposed to be a target to suppress cell proliferation in E. coli, the malarial parasite and tuberculosis. In this manuscript we screened a 70-member library of ATC inhibitors developed against the malarial and tubercular ATCases for inhibitors of the human ATC. Four compounds showed low nanomolar inhibition (IC50 30-120 nM) in an in vitro activity assay. These compounds significantly outperform PALA, which has a triphasic inhibition response under identical conditions, in which significant activity remains at PALA concentrations above 10 μM. Evidence for a druggable allosteric pocket in human ATC is provided by both in vitro enzyme kinetic, homology modeling and in silico docking. These compounds also suppress the proliferation of U2OS osteoblastoma cells by promoting cell cycle arrest in G0/G1 phase. This report provides the first evidence for an allosteric pocket in human ATC, which greatly enhances its druggability and demonstrates the potential of this series in cancer therapy.

• Klíčová slova
• Aspartate Transcarbamoylase, Molecular Docking, Non-competitive Inhibition, Osteosarcoma, Pyrimidine Biosynthesis,
• MeSH
• alosterická regulace účinky léků   MeSH
• antitumorózní látky farmakologie   chemie   chemická syntéza   MeSH
• aspartátkarbamoyltransferasa * antagonisté a inhibitory   metabolismus   chemie   MeSH
• epitelové buňky účinky léků   metabolismus   MeSH
• inhibitory enzymů * farmakologie   chemie   chemická syntéza   MeSH
• knihovny malých molekul chemie   farmakologie   chemická syntéza   MeSH
• léky antitumorózní - screeningové testy MeSH
• lidé MeSH
• molekulární struktura MeSH
• nádorové buněčné linie MeSH
• nádory kostí farmakoterapie   patologie   metabolismus   MeSH
• osteosarkom * farmakoterapie   patologie   metabolismus   MeSH
• proliferace buněk * účinky léků   MeSH
• simulace molekulového dockingu 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
• antitumorózní látky MeSH
• aspartátkarbamoyltransferasa * MeSH
• inhibitory enzymů * MeSH
• knihovny malých molekul MeSH

The goal of this study was to develop novel radioprotective agents targeting the intrinsic apoptotic pathway and thus decreasing the radiation-induced damage. For that purpose, we designed, synthesized and analyzed ten new compounds based on the 1-(4-(2-hydroxyethyl)piperazin-1-yl)-3-phenoxypropan-2-ol leading structure. The cytotoxicity of the newly synthesized substances was tested in vitro on cell lines derived from different progenitor cells by WST-1 proliferation assay. MTT test was utilized to assess half-maximal inhibitory concentrations and maximum tolerated concentrations of novel compounds in A-549 cells. Screening for radioprotective properties was performed using flow-cytometry in MOLT-4 cells exposed to 60Co ionizing gamma radiation. Selected candidates underwent in vivo testing in C57Bl/6 J mice having a positive impact on their immunological status. In summary, we report here promising compounds with radioprotective effect in vivo.

• Klíčová slova
• 1-(2-hydroxyethyl)piperazine derivative, In vitro, Ionizing radiation, Mice, Radioprotection, Synthesis,
• MeSH
• apoptóza účinky léků   MeSH
• knihovny malých molekul chemická syntéza   chemie   farmakologie   MeSH
• lidé MeSH
• molekulární struktura MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• proliferace buněk účinky léků   MeSH
• propanoly chemická syntéza   chemie   farmakologie   MeSH
• radioprotektivní látky chemická syntéza   chemie   farmakologie   MeSH
• simulace molekulového dockingu MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• vztahy mezi strukturou a aktivitou MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• knihovny malých molekul MeSH
• propanoly MeSH
• radioprotektivní látky MeSH

The purpose of this study was to identify new small molecules that possess activity on human toll-like receptor 4 associated with the myeloid differentiation protein 2 (hTLR4/MD2). Following current rational drug design principles, we firstly performed a ligand and structure based virtual screening of more than 130 000 compounds to discover until now unknown class of hTLR4/MD2 modulators that could be used as novel type of immunologic adjuvants. The core of the in silico study was molecular docking of flexible ligands in a partially flexible hTLR4/MD2 receptor model using a peta-flops-scale supercomputer. The most promising substances resulting from this study, related to anthracene-succimide hybrids, were synthesized and tested. The best prepared candidate exhibited 80% of Monophosphoryl Lipid A in vitro agonistic activity in cell lines expressing hTLR4/MD2.

• Klíčová slova
• Adjuvants, Innate immunity system, PRR, TLR4, Virtual screening,
• MeSH
• buněčné linie MeSH
• knihovny malých molekul chemická syntéza   chemie   farmakologie   MeSH
• lidé MeSH
• ligandy MeSH
• molekulární struktura MeSH
• počítačová simulace * MeSH
• preklinické hodnocení léčiv MeSH
• racionální návrh léčiv * MeSH
• simulace molekulového dockingu MeSH
• toll-like receptor 4 antagonisté a inhibitory   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
• knihovny malých molekul MeSH
• ligandy MeSH
• TLR4 protein, human MeSH Prohlížeč
• toll-like receptor 4 MeSH

FOXO transcription factors are critical regulators of cell homeostasis and steer cell death, differentiation and longevity in mammalian cells. By combined pharmacophore-modeling-based in silico and fluorescence polarization-based screening we identified small molecules that physically interact with the DNA-binding domain (DBD) of FOXO3 and modulate the FOXO3 transcriptional program in human cells. The mode of interaction between compounds and the FOXO3-DBD was assessed via NMR spectroscopy and docking studies. We demonstrate that compounds S9 and its oxalate salt S9OX interfere with FOXO3 target promoter binding, gene transcription and modulate the physiologic program activated by FOXO3 in cancer cells. These small molecules prove the druggability of the FOXO-DBD and provide a structural basis for modulating these important homeostasis regulators in normal and malignant cells.

• Klíčová slova
• FOXO transcription factors, biochemistry, cancer biology, chemical biology, docking, drug targeting, human, molecular biophysics, pharmacophore modelling, small compounds, structural biology,
• MeSH
• DNA chemie   genetika   metabolismus   MeSH
• genetická transkripce účinky léků   MeSH
• genový knockdown MeSH
• HEK293 buňky MeSH
• knihovny malých molekul chemie   metabolismus   farmakologie   MeSH
• konformace nukleové kyseliny MeSH
• lidé MeSH
• magnetická rezonanční spektroskopie MeSH
• molekulární modely MeSH
• nádorové buněčné linie MeSH
• promotorové oblasti (genetika) genetika   MeSH
• protein FOXO3 chemie   genetika   metabolismus   MeSH
• proteinové domény MeSH
• simulace molekulového dockingu MeSH
• stanovení celkové genové exprese metody   MeSH
• vazba proteinů MeSH
• vazebná místa genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA MeSH
• FOXO3 protein, human MeSH Prohlížeč
• knihovny malých molekul MeSH
• protein FOXO3 MeSH

The formation of a G-quadruplex motif in the promoter region of the c-MYC protooncogene prevents its expression. Accordingly, G-quadruplex stabilization by a suitable ligand may be a viable approach for anticancer therapy. In our study, we used the 4-(4-methylpiperazin-1-yl)aniline molecule, previously identified as a fragment library screen hit, as a template for the SAR-guided design of a new small library of clickable fragments and subjected them to click reactions, including kinetic target-guided synthesis in the presence of a G-quadruplex forming oligonucleotide Pu24. We tested the clickable fragments and products of click reactions for their G-quadruplex stabilizing activity and determined their mode of binding to the c-MYC G-quadruplex by NMR spectroscopy. The enhanced stabilizing potency of click products in biology assays (FRET, Polymerase extension assay) matched the increased yields of in situ click reactions. In conclusion, we identified the newly synthesized click products of bis-amino derivatives of 4-(4-methylpiperazin-1-yl)aniline as potent stabilizers of c-MYC G-quadruplex, and their further evolution may lead to the development of an efficient tool for cancer treatment.

• Klíčová slova
• G-quadruplex, NMR, anticancer therapy, click chemistry, secondary structures,
• MeSH
• aniliny chemická syntéza   chemie   farmakologie   MeSH
• G-kvadruplexy účinky léků   MeSH
• geny myc genetika   MeSH
• kinetika MeSH
• ligandy MeSH
• simulace molekulární dynamiky MeSH
• syntetická chemie okamžité shody MeSH
• techniky syntetické chemie MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aniline MeSH Prohlížeč
• aniliny MeSH
• ligandy MeSH

Pharmacophore models are widely used for the identification of promising primary hits in compound large libraries. Recent studies have demonstrated that pharmacophores retrieved from protein-ligand molecular dynamic trajectories outperform pharmacophores retrieved from a single crystal complex structure. However, the number of retrieved pharmacophores can be enormous, thus, making it computationally inefficient to use all of them for virtual screening. In this study, we proposed selection of distinct representative pharmacophores by the removal of pharmacophores with identical three-dimensional (3D) pharmacophore hashes. We also proposed a new conformer coverage approach in order to rank compounds using all representative pharmacophores. Our results for four cyclin-dependent kinase 2 (CDK2) complexes with different ligands demonstrated that the proposed selection and ranking approaches outperformed the previously described common hits approach. We also demonstrated that ranking, based on averaged predicted scores obtained from different complexes, can outperform ranking based on scores from an individual complex. All developments were implemented in open-source software pharmd.

• Klíčová slova
• molecular dynamics, pharmacophore, virtual screening,
• MeSH
• cyklin-dependentní kinasa 2 chemie   metabolismus   MeSH
• inhibitory proteinkinas chemie   farmakologie   MeSH
• knihovny malých molekul chemie   farmakologie   MeSH
• lidé MeSH
• ligandy MeSH
• objevování léků metody   MeSH
• počítačová simulace MeSH
• simulace molekulární dynamiky * MeSH
• simulace molekulového dockingu metody   MeSH
• vazba proteinů MeSH
• vazebná místa MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cyklin-dependentní kinasa 2 MeSH
• inhibitory proteinkinas MeSH
• knihovny malých molekul MeSH
• ligandy MeSH

Combinatory strategies using pharmacology and stem cell therapy have emerged due to their potential in the treatment of retinal pigment epithelium (RPE) cell related diseases, and a variety of different stem cell sources have been evaluated both in animal models and in humans. RPE cells derived from human embryonic stem cells (hESCs) and human induced pluripotent cells (hiPSCs) are already in clinical trials, holding great promise for the treatment of age-related macular disease (AMD) and hereditary RPE-related retinal dystrophies. Highly efficient protocol for RPE generations have been developed, but they are still time-consuming and laborious. Areas covered: The authors review RPE related diseases, as well as the known functions of RPE cells in retinal homeostasis. The authors also discuss small molecules that target RPE in vivo as well as in vitro to aid RPE differentiation from pluripotent stem cells clinically. The authors base this review on literature searches performed through PubMed. Expert opinion: Using high-throughput systems, technology will provide the possibility of identifying and optimizing molecules/drugs that could lead to faster and simpler protocols for RPE differentiation. This could be crucial in moving forward to create safer and more efficient RPE-based personalized therapies.

• Klíčová slova
• AMD, Hipscs, RPE differentiation, cell therapy, retinal dystrophies, retinal pigment epithelial cells, small molecules,
• MeSH
• buněčná diferenciace fyziologie   MeSH
• kombinovaná terapie MeSH
• lidé MeSH
• makulární degenerace patofyziologie   terapie   MeSH
• nemoci retiny patofyziologie   terapie   MeSH
• pluripotentní kmenové buňky cytologie   MeSH
• retinální pigmentový epitel cytologie   MeSH
• rychlé screeningové testy MeSH
• transplantace kmenových buněk metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH