NAG-thiazoline is a well-established competitive inhibitor of two physiologically relevant glycosidase families-β-N-acetylhexosaminidases (GH20) and β-N-acetylglucosaminidases (GH84). Based on the different substrate flexibilities of these enzyme groups, we designed and synthesized the 4-deoxy derivative of NAG-thiazoline aiming at the selective inhibition of GH20 β-N-acetylhexosaminidases. One GH84 and two GH20 microbial glycosidases were employed as model enzymes for the inhibition assays. Surprisingly, the new compound 4-deoxy-thiazoline exhibited no activity inhibition with either of the enzyme families of interest. Unlike with the substrates, the 4-hydroxyl group of the inhibitor's sugar ring seems to be crucial for binding the inhibitor to the active sites of these enzymes.
- MeSH
- acetylglukosamin analogy a deriváty chemická syntéza chemie metabolismus MeSH
- Bacteroides enzymologie MeSH
- bakteriální proteiny antagonisté a inhibitory metabolismus MeSH
- beta-N-acetylhexosaminidasy antagonisté a inhibitory metabolismus MeSH
- fungální proteiny antagonisté a inhibitory metabolismus MeSH
- houby enzymologie MeSH
- kinetika MeSH
- Streptomyces enzymologie MeSH
- substrátová specifita MeSH
- thiazoly chemická syntéza chemie metabolismus MeSH
- vazba proteinů MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
NAG-thiazoline is a strong competitive inhibitor of GH20 β-N-acetyl- hexosaminidases and GH84 β-N-acetylglucosaminidases. Here, we focused on the design, synthesis and inhibition potency of a series of new derivatives of NAG-thiazoline modified at the C-6 position. Dimerization of NAG-thiazoline via C-6 attached triazole linkers prepared by click chemistry was employed to make use of multivalency in the inhibition. Novel compounds were tested as potential inhibitors of β-N-acetylhexosaminidases from Talaromyces flavus, Streptomyces plicatus (both GH20) and β-N-acetylglucosaminidases from Bacteroides thetaiotaomicron and humans (both GH84). From the set of newly prepared NAG-thiazoline derivatives, only C-6-azido-NAG-thiazoline displayed inhibition activity towards these enzymes; C-6 triazole-substituted NAG-thiazolines lacked inhibition activity against the enzymes used. Docking of C-6-azido-NAG-thiazoline into the active site of the tested enzymes was performed. Moreover, a stability study with GlcNAc-thiazoline confirmed its decomposition at pH < 6 yielding 2-acetamido-2-deoxy-1-thio-α/β-D-glucopyranoses, which presumably dimerize oxidatively into S-S linked dimers; decomposition products of NAG-thiazoline are void of inhibitory activity.
- MeSH
- acetylglukosamin analogy a deriváty chemická syntéza chemie farmakologie MeSH
- beta-N-acetylhexosaminidasy antagonisté a inhibitory chemie metabolismus MeSH
- glykosidhydrolasy antagonisté a inhibitory chemie metabolismus MeSH
- katalytická doména MeSH
- molekulární konformace MeSH
- molekulární modely MeSH
- stabilita léku MeSH
- thiazoly chemická syntéza chemie farmakologie MeSH
- vazba proteinů MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
On the basis of the highly branched ovomucoid-type undecasaccharide that had been shown previously to be an endogenous ligand for CD69 leukocyte receptor, a systematic investigation of smaller oligosaccharide mimetics was performed based on linear and branched N-acetyl-d-hexosamine homooligomers prepared synthetically using hitherto unexplored reaction schemes. The systematic structure-activity studies revealed the tetrasaccharide GlcNAcbeta1-3(GlcNAcbeta1-4)(GlcNAcbeta1-6)GlcNAc (compound 52) and its alpha-benzyl derivative 49 as the best ligand for CD69 with IC(50) as high as 10(-9) M. This compound thus approaches the affinity of the classical high-affinity neoglycoprotein ligand GlcNAc(23)BSA. Compound 68, GlcNAc tetrasaccharide 52 dimerized through a hydrophilic flexible linker, turned out to be effective in activating CD69(+) lymphocytes. It also proved efficient in enhancing natural killing in vitro, decreasing the growth of tumors in vivo, and activating the CD69(+) tumor infiltrating lymphocytes examined ex vivo. This compound is thus a candidate for carbohydrate-based immunomodulators with promising antitumor potential.
- MeSH
- acetylglukosamin analogy a deriváty chemická syntéza chemie farmakologie MeSH
- aktivace lymfocytů MeSH
- antitumorózní látky chemická syntéza chemie farmakologie MeSH
- buňky NK účinky léků imunologie metabolismus MeSH
- CD antigeny metabolismus MeSH
- diferenciační antigeny T-lymfocytů metabolismus MeSH
- dimerizace MeSH
- imunologické faktory chemická syntéza chemie farmakologie MeSH
- krysa rodu rattus MeSH
- lektinové receptory NK-buněk - podrodina B metabolismus MeSH
- lektiny typu C metabolismus MeSH
- léky antitumorózní - screeningové testy MeSH
- lidé MeSH
- ligandy MeSH
- melanom experimentální imunologie patologie MeSH
- molekulární mimikry MeSH
- molekulární modely MeSH
- molekulární sekvence - údaje MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- oligosacharidy chemická syntéza chemie farmakologie MeSH
- rekombinantní proteiny chemie MeSH
- sacharidové sekvence MeSH
- techniky in vitro MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- publikace stažené z tisku MeSH
