Pharmacological inhibition of cyclin-dependent kinases has emerged as a possible treatment option for various cancer types. We recently identified substituted imidazo[1,2-c]pyrimidin-5(6H)-ones as inhibitors of cyclin-dependent kinase 2 (CDK2). Here, we report the synthesis of derivatives modified at positions 2, 3, 6 or 8 prepared using Suzuki-Miyaura cross-coupling, halogenation, Dimroth-type rearrangement and alkylation as the main synthetic methods. The compounds displayed micro- to submicromolar inhibition of CDK2/cyclin E activity. Binding of the most potent compound 3b to CDK2 was determined using isothermal titration calorimetry. The co-crystal structure of 3b in complex with fully active CDK2 was solved, revealing the binding mode of 3b in the ATP pocket and a hydrogen bonding interaction with hinge region residue Leu83. Evaluation against leukaemia cell lines revealed low cytotoxicity, which is in line with the high selectivity towards CDK2. This study demonstrates that substituted imidazo[1,2-c]pyrimidines can be exploited for future kinase inhibitor development.

• MeSH
• cyklin-dependentní kinasa 2 antagonisté a inhibitory   metabolismus   MeSH
• imidazoly chemie   metabolismus   farmakologie   MeSH
• inhibitory proteinkinas chemická syntéza   metabolismus   farmakologie   MeSH
• kontrolní body buněčného cyklu účinky léků   MeSH
• krystalografie rentgenová MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• proliferace buněk účinky léků   MeSH
• protinádorové látky chemická syntéza   metabolismus   farmakologie   MeSH
• pyrimidiny chemie   metabolismus   farmakologie   MeSH
• simulace molekulární dynamiky MeSH
• vazba proteinů MeSH
• vazebná místa MeSH
• vodíková vazba MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

A side effect of diabetes is formation of glycated proteins and, from them, production of advanced early glycation end products that could determine aberrant immune responses at the systemic level. We investigated a relevant aberrant post-translational modification (PTM) in diabetes based on synthetic peptides modified on the lysine side chain residues with 1-deoxyfructopyranosyl moiety as a possible modification related to glycation. The PTM peptides were used as molecular probes for detection of possible specific autoantibodies developed by diabetic patients. The PDC-E2(167-186) sequence from the pyruvate dehydrogenase complex was selected and tested as a candidate peptide for antibody detection. The structure-based designed type I' β-turn CSF114 peptide was also used as a synthetic scaffold. Twenty-seven consecutive type 1 diabetic patients and 29 healthy controls were recruited for the study. In principle, the 'chemical reverse approach', based on the use of patient sera to screen the synthetic modified peptides, leads to the identification of specific probes able to characterize highly specific autoantibodies as disease biomarkers of autoimmune disorders. Quite surprisingly, both peptides modified with the (1-deoxyfructosyl)-lysine did not lead to significant results. Both IgG and IgM differences between the two populations were not significant. These data can be rationalized considering that i) IgGs in diabetic subjects exhibit a high degree of glycation, leading to decreased functionality; ii) IgGs in diabetic subjects exhibit a privileged response vs proteins containing advanced glycation products (e.g., methylglyoxal, glyoxal, glucosone, hydroimidazolone, dihydroxyimidazolidine) and only a minor one with respect to (1-deoxyfructosyl)-lysine.

• MeSH
• diabetes mellitus 1. typu metabolismus   MeSH
• glykosylace MeSH
• glyoxal metabolismus   MeSH
• imidazoly metabolismus   MeSH
• imunoanalýza MeSH
• ketosy metabolismus   MeSH
• lidé MeSH
• lysin chemie   metabolismus   MeSH
• peptidy chemie   metabolismus   MeSH
• posttranslační úpravy proteinů MeSH
• produkty pokročilé glykace metabolismus   MeSH
• pyruvaldehyd metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Plant cytosolic aldehyde dehydrogenases from family 2 (ALDH2s, EC 1.2.1.3) are non-specific enzymes and participate for example in the metabolism of acetaldehyde or biosynthesis of phenylpropanoids. Plant aminoaldehyde dehydrogenases (AMADHs, ALDH10 family, EC 1.2.1.19) are broadly specific and play an important role in polyamine degradation or production of osmoprotectants. We have tested imidazole and pyrazole carbaldehydes and their alkyl-, allyl-, benzyl-, phenyl-, pyrimidinyl- or thienyl-derivatives as possible substrates of plant ALDH2 and ALDH10 enzymes. Imidazole represents a building block of histidine, histamine as well as certain alkaloids. It also appears in synthetic pharmaceuticals such as imidazole antifungals. Biological compounds containing pyrazole are rare (e.g. pyrazole-1-alanine and pyrazofurin antibiotics) but the ring is often found as a constituent of many synthetic drugs and pesticides. The aim was to evaluate whether aldehyde compounds based on azole heterocycles are oxidized by the enzymes, which would further support their expected role as detoxifying aldehyde scavengers. The analyzed imidazole and pyrazole carbaldehydes were only slowly converted by ALDH10s but well oxidized by cytosolic maize ALDH2 isoforms (particularly by ALDH2C1). In the latter case, the respective Km values were in the range of 10-2000 μmol l-1; the kcat values appeared mostly between 0.1 and 1.0 s-1. The carbaldehyde group at the position 4 of imidazole was oxidized faster than that at the position 2. Such a difference was not observed for pyrazole carbaldehydes. Aldehydes with an aromatic substituent on their heterocyclic ring were oxidized faster than those with an aliphatic substituent. The most efficient of the tested substrates were comparable to benzaldehyde and p-anisaldehyde known as the best aromatic aldehyde substrates of plant cytosolic ALDH2s in vitro.

• MeSH
• aldehyddehydrogenasa metabolismus   MeSH
• aldehydy chemie   metabolismus   MeSH
• hrách setý enzymologie   MeSH
• imidazoly chemie   metabolismus   MeSH
• kukuřice setá enzymologie   MeSH
• molekulární struktura MeSH
• oxidace-redukce MeSH
• pyrazoly chemie   metabolismus   MeSH
• Solanum lycopersicum enzymologie   MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Imidacloprid-urea is the primary imidacloprid soil metabolite, whereas imidacloprid-olefin is the main plant-relevant metabolite and is more toxic to insects than imidacloprid. We artificially contaminated potting soil and used quantitative UHPLC-QqQ-MS/MS to determine the imidacloprid, imidacloprid-olefin and imidacloprid-urea distributions in rapeseed green plant tissues and roots after 4 weeks of exposure. RESULTS: In soil, the imidacloprid/imidacloprid-urea molar ratios decreased similarly after the 250 and 2500 µg kg(-1) imidacloprid treatments. The imidacloprid/imidacloprid-urea molar ratios in the root and soil were similar, whereas in the green plant tissue, imidacloprid-urea increased more than twofold compared with the root. Although imidacloprid-olefin was prevalent in the green plant tissues, with imidacloprid/imidacloprid-olefin molar ratios of 2.24 and 1.47 for the 250 and 2500 µg kg(-1) treatments respectively, it was not detected in the root. However, imidacloprid-olefin was detected in the soil after the 2500 µg kg(-1) imidacloprid treatment. CONCLUSION: Significant proportions of imidacloprid-olefin and imidacloprid-urea in green plant tissues were demonstrated. The greater imidacloprid supply increased the imidacloprid-olefin/imidacloprid molar ratio in the green plant tissues. The absence of imidacloprid-olefin in the root excluded its retransport from leaves. The similar imidacloprid/imidacloprid-urea ratios in the soil and root indicated that the root serves primarily for transporting these substances. © 2016 Society of Chemical Industry.

• MeSH
• alkeny metabolismus   MeSH
• Brassica rapa účinky léků   metabolismus   fyziologie   MeSH
• dusíkaté sloučeniny metabolismus   MeSH
• imidazoly metabolismus   MeSH
• kořeny rostlin účinky léků   metabolismus   MeSH
• látky znečišťující půdu toxicita   MeSH
• močovina metabolismus   MeSH
• opylení MeSH
• půda chemie   MeSH
• Publikační typ
• časopisecké články MeSH

Léčba chronické hepatitidy typu C kombinací přímo působících perorálních virostatik (directly acting antivirals, DAA) má vysokou účinnost (až 100 %), minimum kontraindikací a mimořádně příznivý bezpečnostní profil. Fixní kombinace elbasviru s grazoprevirem (Zepatier) představuje nově dostupnou, vysoce účinnou variantu této léčby určenou pro pacienty infikované virem hepatitidy typu C (HCV) genotypu 1 nebo 4. Elbasvir (EBR) je inhibitor NS5A (nestrukturálního proteinu 5A) druhé vlny 1. generace, grazoprevir (GZR) proteázový inhibitor 2. generace. Léčivý přípravek Zepatier obsahuje 50 mg EBR a 100 mg GZR. Standardní dávkování představuje jedna tableta denně.

Chronic hepatitis C therapy using directly acting antivirals (DAA) has high efficacy (up to 100%), minimum contraindications plus an extraordinarily favorable safety profile. Fixed combination of elbasvir and grazoprevir represents a newly available, highly effective variant of this therapy, indicated for patients infected with hepatitis C virus (HCV), genotypes 1 or 4. Elbasvir (EBR) is a NS5A inhibitor of the second wave of the 1st generation; grazoprevir (GZR) is a protease inhibitor of the 2nd generation. The preparation Zepatier includes 50 mg of EBR and 100 mg of GZR. The standard dose is one tablet daily.

• Klíčová slova
• elbasvir, grazoprevir,
• MeSH
• antivirové látky farmakologie   metabolismus   terapeutické užití   MeSH
• benzofurany farmakologie   metabolismus   terapeutické užití   MeSH
• chinoxaliny farmakologie   metabolismus   terapeutické užití   MeSH
• chronická hepatitida C * farmakoterapie   MeSH
• fixní kombinace léků MeSH
• imidazoly farmakologie   metabolismus   terapeutické užití   MeSH
• inhibitory proteas farmakologie   metabolismus   terapeutické užití   MeSH
• klinická studie jako téma MeSH
• kvantitativní vztahy mezi strukturou a aktivitou MeSH
• lidé MeSH
• nežádoucí účinky léčiv MeSH
• virové nestrukturální proteiny farmakologie   metabolismus   terapeutické užití   MeSH
• Check Tag
• lidé MeSH

• MeSH
• antivirové látky MeSH
• benzofurany farmakokinetika   metabolismus   terapeutické užití   MeSH
• chinoxaliny farmakokinetika   metabolismus   terapeutické užití   MeSH
• chronická hepatitida C diagnóza   farmakoterapie   komplikace   MeSH
• chronická renální insuficience farmakoterapie   komplikace   MeSH
• fixní kombinace léků MeSH
• hodnocení léčiv MeSH
• imidazoly farmakokinetika   metabolismus   terapeutické užití   MeSH
• inhibitory proteas škodlivé účinky   terapeutické užití   MeSH
• komorbidita MeSH
• lidé MeSH
• nežádoucí účinky léčiv epidemiologie   etiologie   MeSH
• Check Tag
• lidé MeSH

Few cases of spontaneously horizontally transferred bacterial genes into plant genomes have been described to date. The occurrence of horizontally transferred genes from the T-DNA of Agrobacterium rhizogenes into the plant genome has been reported in the genus Nicotiana and in the species Linaria vulgaris. Here we compare patterns of evolution in one of these genes (a gene encoding mikimopine synthase, mis) following three different events of horizontal gene transfer (HGT). As this gene plays an important role in Agrobacterium, and there are known cases showing that genes from pathogens can acquire plant protection function, we hypothesised that in at least some of the studied species we will find signs of selective pressures influencing mis sequence. The mikimopine synthase (mis) gene evolved in a different manner in the branch leading to Nicotiana tabacum and N. tomentosiformis, in the branch leading to N. glauca and in the genus Linaria. Our analyses of the genus Linaria suggest that the mis gene began to degenerate soon after the HGT. In contrast, in the case of N. glauca, the mis gene evolved under significant selective pressures. This suggests a possible role of mikimopine synthase in current N. glauca and its ancestor(s). In N. tabacum and N. tomentosiformis, the mis gene has a common frameshift mutation that disrupted its open reading frame. Interestingly, our results suggest that in spite of the frameshift, the mis gene could evolve under selective pressures. This sequence may still have some regulatory role at the RNA level as suggested by coverage of this sequence by small RNAs in N. tabacum.

• MeSH
• Agrobacterium enzymologie   genetika   MeSH
• bakteriální proteiny klasifikace   genetika   metabolismus   MeSH
• druhová specificita MeSH
• fylogeneze MeSH
• imidazoly metabolismus   MeSH
• interakce hostitele a patogenu genetika   MeSH
• Linaria genetika   mikrobiologie   MeSH
• molekulární evoluce MeSH
• molekulární sekvence - údaje MeSH
• oxidoreduktasy působící na CH-NH vazby klasifikace   genetika   metabolismus   MeSH
• posunová mutace MeSH
• přenos genů horizontální * MeSH
• pyridiny metabolismus   MeSH
• regulace genové exprese enzymů MeSH
• sekvence nukleotidů MeSH
• selekce (genetika) MeSH
• tabák klasifikace   genetika   mikrobiologie   MeSH
• transformace genetická MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• Klíčová slova
• adherence k SPC, studie ATLAS, hodnocení bezpečnosti,
• MeSH
• adherence k farmakoterapii * psychologie   statistika a číselné údaje   MeSH
• bisfosfonáty * aplikace a dávkování   kontraindikace   metabolismus   škodlivé účinky   terapeutické užití   MeSH
• hodnotící studie jako téma MeSH
• imidazoly aplikace a dávkování   kontraindikace   metabolismus   škodlivé účinky   terapeutické užití   MeSH
• inhibitory kostní resorpce terapeutické užití   MeSH
• klinické zkoušky jako téma MeSH
• kyselina zoledronová MeSH
• lidé středního věku MeSH
• lidé MeSH
• osteoporotické fraktury * etiologie   prevence a kontrola   MeSH
• postmenopauzální osteoporóza diagnóza   terapie   MeSH
• statistika jako téma MeSH
• vápník MeSH
• vitamin D fyziologie   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH

MDM2 is a multidomain protein that functions as an E3 ubiquitin ligase, transcription repressor, mRNA-binding protein, translation factor, and molecular chaperone. The small molecule Nutlin-3 has been engineered to bind to the N-terminal hydrophobic pocket domain of MDM2. This binding of Nutlin-3 has two consequences: (i) antagonistic effects through competitive disruption of the MDM2-p53 complex and (ii) agonist effects that allosterically stabilize MDM2 protein-protein interactions that increase p53 ubiquitination as well as nucleophosmin deoligomerization. We present a methodology using a hydrogen/deuterium (H/D) exchange platform that measures Nutlin-3 binding to the N-terminal domain of MDM2 (MDM2(1-126)) in order to begin to develop dynamic assays that evaluate MDM2 allostery. In order to localize the regions in MDM2 being suppressed by Nutlin-3, MDM2 was incubated with the ligand and H/D amide exchange was measured after pepsin digestion. One dynamic segment containing amino acids 55-60 exhibited slower deuterium exchange after Nutlin-3 binding, reflecting ligand binding within the hydrophobic pocket. However, another dominant suppression of H/D exchange was observed in a motif from amino acids 103-107 that reflects surface hydrophobic residues surrounding the hydrophobic pocket of MDM2. In order to explore the consequences of this latter Nutlin-3 interaction site on MDM2, the Y104G and L107G mutant series was constructed. The MDM2(Y104G) and MDM2(L107G) mutants were fully active in p53 binding. However, the authentic p53-derived peptide:MDM2(Y104G) complex exhibited partial resistance to Nutlin-3 inhibition, while the p53-mimetic 12.1 peptide:MDM2(Y104G) complex retained normal Nutlin-3 responsiveness. These data reveal the existence of a second functional Nutlin-3-binding site in a surface hydrophobic patch of MDM2, flanking the hydrophobic pocket. This reveals two modes of peptide binding by MDM2 and highlights the utility of H/D exchange as an assay for measuring allosteric effects in MDM2.

• MeSH
• alosterické místo MeSH
• imidazoly chemie   metabolismus   MeSH
• lidé MeSH
• molekulární modely MeSH
• molekulární sekvence - údaje MeSH
• peptidové fragmenty MeSH
• piperaziny chemie   metabolismus   MeSH
• protoonkogenní proteiny c-mdm2 chemie   metabolismus   MeSH
• sekvence aminokyselin MeSH
• vodík-deuteriová výměna metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

One of the key functions of nitric oxide (NO) in human is to dilate blood vessels. We tested glycerol trinitrate (GTN) and other well-known NO donors together with those bearing a >C=N-OH group for possible conversion to NO (or nitrites, respectively) by diaphorase (DP) and lipoamide dehydrogenase (LAD). Both, DP and LAD were unable to convert formamidoxime (FAM), acetone oxime (AC), acetohydroxamic acid (AHA) and Nomega-hydroxy-L-arginine (L-NOHA). On the other hand, we observed good conversion of GTN without the requirement of superoxide anion. However, superoxide anion participated to a varying extent in the conversion of other donors (formaldoxime (FAL), acetaldoxime (AO), nitroprusside (NP), S-nitrosoglutathione (SNOG), S-nitroso-N-acetylpenicillamine (SNAP) and hydroxylamine (HA)). All DP- and LAD-mediated reactions were inhibited by diphenyleneiodonium chloride (DPI), (an inhibitor of flavine enzymes), in a concentration-dependent manner. For these inhibition reactions we determined Ki and IC50 values. In addition, we found that conversion of SNOG was significantly accelerated by glutathione reductase (GTR). Like with DP, 2-phenyl- 4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO) was reduced also by LAD and thioredoxin reductase (TRR). In summary, we found that LAD significantly accelerates the conversion of a defined subset of NO donors to NO, especially GTN, and eliminates the NO scavenging effect of PTIO.

• MeSH
• biokatalýza MeSH
• Clostridium kluyveri enzymologie   MeSH
• cyklické N-oxidy metabolismus   MeSH
• dihydrolipoamiddehydrogenasa antagonisté a inhibitory   metabolismus   MeSH
• elektronová paramagnetická rezonance MeSH
• imidazoly metabolismus   MeSH
• kinetika MeSH
• oniové sloučeniny farmakologie   MeSH
• oxid dusnatý metabolismus   MeSH
• oxidace-redukce MeSH
• scavengery volných radikálů metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH