TY  - JOUR
AB  - We report for the first time Antibody-Drug-Conjugates (ADCs) containing human (h) Carbonic Anhydrase (CA; EC 4.2.1.1) directed Monoclonal Antibodies (MAbs) linked to low molecular weight inhibitors of the same enzymes by means of hydrophilic peptide spacers. In agreement with the incorporated CA directed MAb fragments, in vitro inhibition data of the obtained ADCs showed sub-nanomolar K-I values for the tumour associated CAs IX and XII which were up to 10-fold more potent when compared to the corresponding unconjugated MAbs. In addition, the introduction of the CA inhibitor (CAI) benzenesulfonamide allowed the ADCs to potently inhibit the housekeeping tumoral off-target human CA II isoform. Such results are supporting the definition of an unprecedented reported class of ADCs able to hit simultaneously multiple hCAs physiologically cooperative in maintaining altered cellular metabolic pathways, and therefore ideal for the treatment of chronic diseases such as cancers and inflammation diseases.
AU  - Testa, C.*
AU  - Papini, A.M.*
AU  - Zeidler, R.
AU  - Vullo, D.*
AU  - Carta, F.*
AU  - Supuran, C.T.*
AU  - Rovero, P.*
C1  - 64154
C2  - 52094
CY  - 2-4 Park Square, Milton Park, Abingdon Or14 4rn, Oxon, England
SP  - 592-596
TI  - First studies on tumor associated carbonic anhydrases IX and XII monoclonal antibodies conjugated to small molecule inhibitors.
JO  - J. Enzyme Inhib. Med. Chem.
VL  - 37
IS  - 1
PB  - Taylor & Francis Ltd
PY  - 2022
SN  - 1475-6366
ER  - 

TY  - JOUR
AB  - Mirolysin is a secretory protease of Tannerella forsythia, a member of the dysbiotic oral microbiota responsible for periodontitis. In this study, we show that mirolysin latency is achieved by a "cysteine-switch" mechanism exerted by Cys23 in the N-terminal profragment. Mutation of Cys23 shortened the time needed for activation of the zymogen from several days to 5 min. The mutation also decreased the thermal stability and autoproteolysis resistance of promirolysin. Mature mirolysin is a thermophilic enzyme and shows optimal activity at 65 °C. Through NMR-based fragment screening, we identified a small molecule (compound (cpd) 9) that blocks promirolysin maturation and functions as a competitive inhibitor (Ki = 3.2 µM), binding to the S1' subsite of the substrate-binding pocket. Cpd 9 shows superior specificity and does not interact with other T. forsythia proteases or Lys/Arg-specific proteases.
AU  - Zak, K.M.
AU  - Bostock, M.J.
AU  - Waligorska, I.*
AU  - Thøgersen, I.B.*
AU  - Enghild, J.J.*
AU  - Popowicz, G.M.
AU  - Grudnik, P.*
AU  - Potempa, J.S.*
AU  - Ksiazek, M.*
C1  - 62432
C2  - 50867
CY  - 2-4 Park Square, Milton Park, Abingdon Or14 4rn, Oxon, England
SP  - 1267-1281
TI  - Latency, thermal stability, and identification of an inhibitory compound of mirolysin, a secretory protease of the human periodontopathogen Tannerella forsythia.
JO  - J. Enzyme Inhib. Med. Chem.
VL  - 36
IS  - 1
PB  - Taylor & Francis Ltd
PY  - 2021
SN  - 1475-6366
ER  - 

TY  - JOUR
AB  - Carbonic anhydrase 9 (CA9) and carbonic anhydrase 12 (CA12) were proposed as potential targets for cancer therapy more than 20 years ago. However, to date, there are only very few antibodies that have been described to specifically target CA9 and CA12 and also block the enzymatic activity of their targets. One of the early stage bottlenecks in identifying CA9- and CA12-inhibiting antibodies has been the lack of a high-throughput screening system that would allow for rapid assessment of inhibition of the targeted carbon dioxide hydratase activity of carbonic anhydrases. In this study, we show that measuring the esterase activity of carbonic anhydrase offers a robust and inexpensive screening method for identifying antibody candidates that block both hydratase and esterase activities of carbonic anhydrase's. To our knowledge, this is the first implementation of a facile surrogate-screening assay to identify potential therapeutic antibodies that block the clinically relevant hydratase activity of carbonic anhydrases.
AU  - Uda, N.R.*
AU  - Seibert, V.*
AU  - Stenner-Liewen, F.*
AU  - Müller, P.*
AU  - Herzig, P.*
AU  - Gondi, G.*
AU  - Zeidler, R.
AU  - van Dijk, M.*
AU  - Zippelius, A.*
AU  - Renner, C.*
C1  - 44445
C2  - 36844
CY  - Abingdon
SP  - 955-960
TI  - Esterase activity of carbonic anhydrases serves as surrogate for selecting antibodies blocking hydratase activity.
JO  - J. Enzyme Inhib. Med. Chem.
VL  - 30
IS  - 6
PB  - Taylor & Francis Ltd
PY  - 2015
SN  - 1475-6366
ER  -