TY  - JOUR
A1  - Klein, Philipp
A1  - Johe, Patrick
A1  - Wagner, Annika
A1  - Jung, Sascha
A1  - Kühlborn, Jonas
A1  - Barthels, Fabian
A1  - Tenzer, Stefan
A1  - Distler, Ute
A1  - Waigel, Waldemar
A1  - Engels, Bernd
A1  - Hellmich, Ute A.
A1  - Opatz, Till
A1  - Schirmeister, Tanja
T1  - New cysteine protease inhibitors: electrophilic (het)arenes and unexpected prodrug identification for the Trypanosoma protease rhodesain
JF  - Molecules
N2  - Electrophilic (het)arenes can undergo reactions with nucleophiles yielding π- or Meisenheimer (σ-) complexes or the products of the S\(_N\)Ar addition/elimination reactions. Such building blocks have only rarely been employed for the design of enzyme inhibitors. Herein, we demonstrate the combination of a peptidic recognition sequence with such electrophilic (het)arenes to generate highly active inhibitors of disease-relevant proteases. We further elucidate an unexpected mode of action for the trypanosomal protease rhodesain using NMR spectroscopy and mass spectrometry, enzyme kinetics and various types of simulations. After hydrolysis of an ester function in the recognition sequence of a weakly active prodrug inhibitor, the liberated carboxylic acid represents a highly potent inhibitor of rhodesain (K\(_i\) = 4.0 nM). The simulations indicate that, after the cleavage of the ester, the carboxylic acid leaves the active site and re-binds to the enzyme in an orientation that allows the formation of a very stable π-complex between the catalytic dyad (Cys-25/His-162) of rhodesain and the electrophilic aromatic moiety. The reversible inhibition mode results because the S\(_N\)Ar reaction, which is found in an alkaline solvent containing a low molecular weight thiol, is hindered within the enzyme due to the presence of the positively charged imidazolium ring of His-162. Comparisons between measured and calculated NMR shifts support this interpretation
KW  - cysteine protease
KW  - rhodesain
KW  - electrophilic (het)arene
KW  - nucleophilic aromatic substitution
KW  - Meisenheimer complex
KW  - π-complex
KW  - prodrug
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-203380
SN  - 1420-3049
VL  - 25
IS  - 6
ER  - 
TY  - JOUR
A1  - Klein, Philipp
A1  - Barthels, Fabian
A1  - Johe, Patrick
A1  - Wagner, Annika
A1  - Tenzer, Stefan
A1  - Distler, Ute
A1  - Le, Thien Anh
A1  - Schmid, Paul
A1  - Engel, Volker
A1  - Engels, Bernd
A1  - Hellmich, Ute A.
A1  - Opatz, Till
A1  - Schirmeister, Tanja
T1  - Naphthoquinones as covalent reversible inhibitors of cysteine proteases — studies on inhibition mechanism and kinetics
JF  - Molecules
N2  - The facile synthesis and detailed investigation of a class of highly potent protease inhibitors based on 1,4-naphthoquinones with a dipeptidic recognition motif (HN-l-Phe-l-Leu-OR) in the 2-position and an electron-withdrawing group (EWG) in the 3-position is presented. One of the compound representatives, namely the acid with EWG = CN and with R = H proved to be a highly potent rhodesain inhibitor with nanomolar affinity. The respective benzyl ester (R = Bn) was found to be hydrolyzed by the target enzyme itself yielding the free acid. Detailed kinetic and mass spectrometry studies revealed a reversible covalent binding mode. Theoretical calculations with different density functionals (DFT) as well as wavefunction-based approaches were performed to elucidate the mode of action.
KW  - protease
KW  - rhodesain
KW  - covalent reversible inhibition
KW  - 1,4-naphthoquinone
KW  - nucleophilic addition
KW  - prodrug
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-203791
SN  - 1420-3049
VL  - 25
IS  - 9
ER  - 
TY  - JOUR
A1  - Barthels, Fabian
A1  - Marincola, Gabriella
A1  - Marciniak, Tessa
A1  - Konhäuser, Matthias
A1  - Hammerschmidt, Stefan
A1  - Bierlmeier, Jan
A1  - Distler, Ute
A1  - Wich, Peter R.
A1  - Tenzer, Stefan
A1  - Schwarzer, Dirk
A1  - Ziebuhr, Wilma
A1  - Schirmeister, Tanja
T1  - Asymmetric Disulfanylbenzamides as Irreversible and Selective Inhibitors of Staphylococcus aureus Sortase A
JF  - ChemMedChem
N2  - Staphylococcus aureus is one of the most frequent causes of nosocomial and community‐acquired infections, with drug‐resistant strains being responsible for tens of thousands of deaths per year. S. aureus sortase A inhibitors are designed to interfere with virulence determinants. We have identified disulfanylbenzamides as a new class of potent inhibitors against sortase A that act by covalent modification of the active‐site cysteine. A broad series of derivatives were synthesized to derive structure‐activity relationships (SAR). In vitro and in silico methods allowed the experimentally observed binding affinities and selectivities to be rationalized. The most active compounds were found to have single‐digit micromolar Ki values and caused up to a 66 % reduction of S. aureus fibrinogen attachment at an effective inhibitor concentration of 10 μM. This new molecule class exhibited minimal cytotoxicity, low bacterial growth inhibition and impaired sortase‐mediated adherence of S. aureus cells.
KW  - antibiotics
KW  - biofilm
KW  - drug design
KW  - sortase A
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-214581
VL  - 15
IS  - 10
SP  - 839
EP  - 850
ER  -