| TYKVART, Jan, Václav NAVRÁTIL, Michael KUGLER, Pavel ŠÁCHA, Jiří SCHIMER, Anna HLAVÁČKOVÁ, Lukáš TENORA, Jitka ZEMANOVÁ, Jitka DEJMEK, Vlastimil KRÁL, Milan POTÁČEK, Pavel MAJER, Ullrich JAHN, Jiří BRYNDA, Pavlína ŘEZÁČOVÁ a Jan KONVALINKA. Identification of Novel Carbonic Anhydrase IX Inhibitors Using High-Throughput Screening of Pooled Compound Libraries by DNA-Linked Inhibitor Antibody Assay (DIANA). SLAS Discovery. SAGE Publications Inc, 2020, roč. 25, č. 9, s. 1026-1037. ISSN 2472-5552. Dostupné z: https://dx.doi.org/10.1177/2472555220918836. |
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@article{1694598,
author = {Tykvart, Jan and Navrátil, Václav and Kugler, Michael and Šácha, Pavel and Schimer, Jiří and Hlaváčková, Anna and Tenora, Lukáš and Zemanová, Jitka and Dejmek, Jitka and Král, Vlastimil and Potáček, Milan and Majer, Pavel and Jahn, Ullrich and Brynda, Jiří and Řezáčová, Pavlína and Konvalinka, Jan},
article_number = {9},
doi = {http://dx.doi.org/10.1177/2472555220918836},
keywords = {high-throughput screening; carbonic anhydrase IX; drug discovery; DNA-linked inhibitor antibody assay},
language = {eng},
issn = {2472-5552},
journal = {SLAS Discovery},
title = {Identification of Novel Carbonic Anhydrase IX Inhibitors Using High-Throughput Screening of Pooled Compound Libraries by DNA-Linked Inhibitor Antibody Assay (DIANA)},
url = {https://doi.org/10.1177/2472555220918836},
volume = {25},
year = {2020}
}
TY - JOUR
ID - 1694598
AU - Tykvart, Jan - Navrátil, Václav - Kugler, Michael - Šácha, Pavel - Schimer, Jiří - Hlaváčková, Anna - Tenora, Lukáš - Zemanová, Jitka - Dejmek, Jitka - Král, Vlastimil - Potáček, Milan - Majer, Pavel - Jahn, Ullrich - Brynda, Jiří - Řezáčová, Pavlína - Konvalinka, Jan
PY - 2020
TI - Identification of Novel Carbonic Anhydrase IX Inhibitors Using High-Throughput Screening of Pooled Compound Libraries by DNA-Linked Inhibitor Antibody Assay (DIANA)
JF - SLAS Discovery
VL - 25
IS - 9
SP - 1026-1037
EP - 1026-1037
PB - SAGE Publications Inc
SN - 24725552
KW - high-throughput screening
KW - carbonic anhydrase IX
KW - drug discovery
KW - DNA-linked inhibitor antibody assay
UR - https://doi.org/10.1177/2472555220918836
L2 - https://doi.org/10.1177/2472555220918836
N2 - The DNA-linked inhibitor antibody assay (DIANA) has been recently validated for ultrasensitive enzyme detection and for quantitative evaluation of enzyme inhibitor potency. Here we present its adaptation for high-throughput screening of human carbonic anhydrase IX (CAIX), a promising drug and diagnostic target. We tested DIANA's performance by screening a unique compound collection of 2816 compounds consisting of lead-like small molecules synthesized at the Institute of Organic Chemistry and Biochemistry (IOCB) Prague (“IOCB library”). Additionally, to test the robustness of the assay and its potential for upscaling, we screened a pooled version of the IOCB library. The results from the pooled screening were in agreement with the initial nonpooled screen with no lost hits and no false positives, which shows DIANA’s potential to screen more than 100,000 compounds per day.All DIANA screens showed a high signal-to-noise ratio with a Z factor of >0.89. The DIANA screen identified 13 compounds with Ki values equal to or better than 10 uM. All retested hits were active also in an orthogonal enzymatic assay showing zero false positives. However, further biophysical validation of identified hits revealed that the inhibition activity of several hits was caused by a single highly potent CAIX inhibitor, being present as a minor impurity. This finding eventually led us to the identification of three novel CAIX inhibitors from the screen. We confirmed the validity of these compounds by elucidating their mode of binding into the CAIX active site by x-ray crystallography.
ER -
TYKVART, Jan, Václav NAVRÁTIL, Michael KUGLER, Pavel ŠÁCHA, Jiří SCHIMER, Anna HLAVÁČKOVÁ, Lukáš TENORA, Jitka ZEMANOVÁ, Jitka DEJMEK, Vlastimil KRÁL, Milan POTÁČEK, Pavel MAJER, Ullrich JAHN, Jiří BRYNDA, Pavlína ŘEZÁČOVÁ a Jan KONVALINKA. Identification of Novel Carbonic Anhydrase IX Inhibitors Using High-Throughput Screening of Pooled Compound Libraries by DNA-Linked Inhibitor Antibody Assay (DIANA). \textit{SLAS Discovery}. SAGE Publications Inc, 2020, roč.~25, č.~9, s.~1026-1037. ISSN~2472-5552. Dostupné z: https://dx.doi.org/10.1177/2472555220918836.
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