Počet záznamů: 1
Identification of Novel Carbonic Anhydrase IX Inhibitors Using High-Throughput Screening of Pooled Compound Libraries by DNA-Linked Inhibitor Antibody Assay (DIANA)
- 1.
SYSNO ASEP 0525049 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Identification of Novel Carbonic Anhydrase IX Inhibitors Using High-Throughput Screening of Pooled Compound Libraries by DNA-Linked Inhibitor Antibody Assay (DIANA) Tvůrce(i) Tykvart, Jan (UOCHB-X) RID, ORCID
Navrátil, Václav (UOCHB-X) RID, ORCID
Kugler, Michael (UOCHB-X) RID, ORCID
Šácha, Pavel (UOCHB-X) RID, ORCID
Schimer, Jiří (UOCHB-X) RID, ORCID
Hlavačková, Anna (UOCHB-X)
Tenora, Lukáš (UOCHB-X) ORCID
Zemanová, Jitka (UOCHB-X)
Dejmek, Milan (UOCHB-X) RID, ORCID
Král, V. (CZ)
Potáček, M. (CZ)
Majer, Pavel (UOCHB-X)
Jahn, Ullrich (UOCHB-X) ORCID
Brynda, Jiří (UOCHB-X) RID, ORCID
Řezáčová, Pavlína (UOCHB-X) RID, ORCID
Konvalinka, Jan (UOCHB-X) RID, ORCIDZdroj.dok. SLAS Discovery. - : Elsevier - ISSN 2472-5552
Roč. 25, č. 9 (2020), s. 1026-1037Poč.str. 12 s. Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova high-throughput screening ; carbonic anhydrase IX ; drug discovery ; DNA-linked inhibitor antibody assay Vědní obor RIV CE - Biochemie Obor OECD Biochemistry and molecular biology CEP GA19-10280S GA ČR - Grantová agentura ČR GA17-14510S GA ČR - Grantová agentura ČR EF16_019/0000729 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy LO1304 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy Způsob publikování Omezený přístup Institucionální podpora UOCHB-X - RVO:61388963 UT WOS 000535984500001 EID SCOPUS 85085386173 DOI 10.1177/2472555220918836 Anotace 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 µM. 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. Pracoviště Ústav organické chemie a biochemie Kontakt asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Jana Procházková, Tel.: 220 183 418 Rok sběru 2021 Elektronická adresa https://journals.sagepub.com/doi/10.1177/2472555220918836
Počet záznamů: 1