Fluorescent A2A and A3 adenosine receptor antagonists as flow cytometry probes
© 2022. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply..
Adenosine receptor (AR) ligands are being developed for metabolic, cardiovascular, neurological, and inflammatory diseases and cancer. The ease of drug discovery is contingent on the availability of pharmacological tools. Fluorescent antagonist ligands for the human A2A and A3ARs were synthesized using two validated pharmacophores, 1,3-dipropyl-8-phenylxanthine and triazolo[1,5-c]quinazolin-5-yl)amine, which were coupled to eight reporter fluorophores: AlexaFluor, JaneliaFluor (JF), cyanine, and near infrared (NIR) dyes. The conjugates were first screened using radioligand binding in HEK293 cells expressing one of the three AR subtypes. The highest affinities at A2AAR were Ki 144-316 nM for 10, 12, and 19, and at A3AR affinity of Ki 21.6 nM for 19. Specific binding of JF646 conjugate MRS7774 12 to the HEK293 cell surface A2AAR was imaged using confocal microscopy. Compound 19 MRS7535, a triazolo[1,5-c]quinazolin-5-yl)amine containing a Sulfo-Cy7 NIR dye, was suitable for A3AR characterization in whole cells by flow cytometry (Kd 11.8 nM), and its bitopic interaction mode with an A3AR homology model was predicted. Given its affinity and selectivity (11-fold vs. A2AAR, ~ 50-fold vs. A1AR and A2BAR) and a good specific-to-nonspecific binding ratio, 19 could be useful for live cell or potentially a diagnostic in vivo NIR imaging tool and/or therapy targeting the A3AR.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2023 |
|---|---|
| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:19 |
|---|---|
| Enthalten in: |
Purinergic signalling - 19(2023), 3 vom: 25. Sept., Seite 565-578 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Toti, Kiran S [VerfasserIn] |
|---|
| Links: |
|---|
| Anmerkungen: |
Date Completed 23.10.2023 Date Revised 10.02.2024 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1007/s11302-022-09873-3 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM342088971 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM342088971 | ||
| 003 | DE-627 | ||
| 005 | 20240210232433.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231226s2023 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1007/s11302-022-09873-3 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1286.xml |
| 035 | |a (DE-627)NLM342088971 | ||
| 035 | |a (NLM)35687212 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Toti, Kiran S |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Fluorescent A2A and A3 adenosine receptor antagonists as flow cytometry probes |
| 264 | 1 | |c 2023 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 23.10.2023 | ||
| 500 | |a Date Revised 10.02.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2022. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply. | ||
| 520 | |a Adenosine receptor (AR) ligands are being developed for metabolic, cardiovascular, neurological, and inflammatory diseases and cancer. The ease of drug discovery is contingent on the availability of pharmacological tools. Fluorescent antagonist ligands for the human A2A and A3ARs were synthesized using two validated pharmacophores, 1,3-dipropyl-8-phenylxanthine and triazolo[1,5-c]quinazolin-5-yl)amine, which were coupled to eight reporter fluorophores: AlexaFluor, JaneliaFluor (JF), cyanine, and near infrared (NIR) dyes. The conjugates were first screened using radioligand binding in HEK293 cells expressing one of the three AR subtypes. The highest affinities at A2AAR were Ki 144-316 nM for 10, 12, and 19, and at A3AR affinity of Ki 21.6 nM for 19. Specific binding of JF646 conjugate MRS7774 12 to the HEK293 cell surface A2AAR was imaged using confocal microscopy. Compound 19 MRS7535, a triazolo[1,5-c]quinazolin-5-yl)amine containing a Sulfo-Cy7 NIR dye, was suitable for A3AR characterization in whole cells by flow cytometry (Kd 11.8 nM), and its bitopic interaction mode with an A3AR homology model was predicted. Given its affinity and selectivity (11-fold vs. A2AAR, ~ 50-fold vs. A1AR and A2BAR) and a good specific-to-nonspecific binding ratio, 19 could be useful for live cell or potentially a diagnostic in vivo NIR imaging tool and/or therapy targeting the A3AR | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Intramural | |
| 650 | 4 | |a Adenosine receptor | |
| 650 | 4 | |a Antagonist | |
| 650 | 4 | |a Drug discovery | |
| 650 | 4 | |a Flow cytometry | |
| 650 | 4 | |a Fluorescent ligands | |
| 650 | 4 | |a Receptor binding | |
| 650 | 7 | |a Purinergic P1 Receptor Antagonists |2 NLM | |
| 650 | 7 | |a Fluorescent Dyes |2 NLM | |
| 650 | 7 | |a Amines |2 NLM | |
| 650 | 7 | |a Receptor, Adenosine A3 |2 NLM | |
| 650 | 7 | |a Receptor, Adenosine A2A |2 NLM | |
| 650 | 7 | |a Adenosine A2 Receptor Antagonists |2 NLM | |
| 700 | 1 | |a Campbell, Ryan G |e verfasserin |4 aut | |
| 700 | 1 | |a Lee, Hobin |e verfasserin |4 aut | |
| 700 | 1 | |a Salmaso, Veronica |e verfasserin |4 aut | |
| 700 | 1 | |a Suresh, R Rama |e verfasserin |4 aut | |
| 700 | 1 | |a Gao, Zhan-Guo |e verfasserin |4 aut | |
| 700 | 1 | |a Jacobson, Kenneth A |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Purinergic signalling |d 2004 |g 19(2023), 3 vom: 25. Sept., Seite 565-578 |w (DE-627)NLM160620074 |x 1573-9546 |7 nnns |
| 773 | 1 | 8 | |g volume:19 |g year:2023 |g number:3 |g day:25 |g month:09 |g pages:565-578 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1007/s11302-022-09873-3 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 19 |j 2023 |e 3 |b 25 |c 09 |h 565-578 | ||