Rhomboids are intramembrane serine proteases with diverse physiological functions in organisms ranging from archaea to humans. Crystal structure analysis has provided a detailed understanding of the catalytic mechanism, and rhomboids have been implicated in various disease contexts. Unfortunately, the design of specific rhomboid inhibitors has lagged behind, and previously described small molecule inhibitors displayed insufficient potency and/or selectivity. Using a computer-aided approach, we focused on the discovery of novel scaffolds with reduced liabilities and the possibility for broad structural variations. Docking studies with the E. coli rhomboid GlpG indicated that 2-styryl substituted benzoxazinones might comprise novel rhomboid inhibitors. Protease in vitro assays confirmed activity of 2-styryl substituted benzoxazinones against GlpG but not against the soluble serine protease α-chymotrypsin. Furthermore, mass spectrometry analysis demonstrated covalent modification of the catalytic residue Ser201, corroborating the predicted mechanism of inhibition and the formation of an acyl enzyme intermediate. In conclusion, 2-styryl substituted benzoxazinones are a novel rhomboid inhibitor scaffold with ample opportunity for optimization.
- MeSH
- benzoxaziny chemická syntéza chemie MeSH
- chymotrypsin chemie MeSH
- DNA vazebné proteiny antagonisté a inhibitory chemie genetika MeSH
- Drosophila chemie MeSH
- endopeptidasy chemie genetika MeSH
- enzymatické testy MeSH
- Escherichia coli enzymologie MeSH
- inhibitory serinových proteinas chemická syntéza chemie MeSH
- katalytická doména MeSH
- lidé MeSH
- membránové proteiny antagonisté a inhibitory chemie genetika MeSH
- mutace MeSH
- objevování léků MeSH
- proteiny Drosophily metabolismus MeSH
- proteiny z Escherichia coli antagonisté a inhibitory chemie genetika MeSH
- serin chemie MeSH
- simulace molekulového dockingu MeSH
- skot MeSH
- styreny chemická syntéza chemie MeSH
- transformující růstový faktor alfa metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
In this study, series of ring-substituted 2-styrylquinazolin-4(3H)-one and 4-chloro-2-styrylquinazoline derivatives were prepared. The syntheses of the discussed compounds are presented. The compounds were analyzed by RP-HPLC to determine lipophilicity. They were tested for their inhibitory activity on photosynthetic electron transport (PET) in spinach (Spinacia oleracea L.) chloroplasts. Primary in vitro screening of the synthesized compounds was also performed against four mycobacterial strains and against eight fungal strains. Several compounds showed biological activity comparable with or higher than that of the standard isoniazid. It was found that the electronic properties of the R substituent, and not the total lipophilicity of the compound, were decisive for the photosynthesis-inhibiting activity of tested compounds.
- MeSH
- antituberkulotika farmakologie chemická syntéza chemie MeSH
- atypické mykobakteriální infekce farmakoterapie MeSH
- bakteriální pneumonie farmakoterapie mikrobiologie MeSH
- chinazoliny farmakologie chemická syntéza chemie MeSH
- chloroplasty účinky léků MeSH
- fotosyntéza účinky léků MeSH
- lidé MeSH
- netuberkulózní mykobakterie účinky léků MeSH
- Spinacia oleracea účinky léků MeSH
- styreny farmakologie chemická syntéza MeSH
- transport elektronů účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH