Rhodol-based thallium sensors for high-throughput screening of potassium channels
AssigneeVanderbilt University
InventorsCharles David Weaver, Brendan F. Dutter, Gary A. Sulikowski
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Abstract
Provided are novel rhodol-based compounds, the preparation method thereof, and use thereof in Tl+ flux assays. The fluorescent thallium ion sensor compounds have a rhodol fluorophore attached to an amino dicarboxylic acid metal binding moiety. The compounds are prepared in the “pro-dye” form, with the rhodol oxygen and the carboxylic acids of the metal binding unit masked by protecting groups. The disclosed compounds may be used as more red-shifted and less pH-sensitive variants of Thallos.
Claims18
A compound of the following formula,
- or a salt thereof,
- wherein:
- Z is O, CRxRy, SiRxRy, or SO2;
- R1 is C1-10alkyl-C(O)—;
- —NR2R3 is —NHC(O)CH3, —N(CH3)2, —N(CH2CH3)2, —N(CH2CH2CH3)2, —N(CH2CH2CH2CH3)2, —NH-cyclohexyl, —NH—CH2CH2CH2CH3, —NH-phenyl, azetidin-1-yl, 3,3-difluoroazetidin-1-yl, pyrrolidin-1-yl, or piperidin-1-yl;
- R4 is C1-10alkyl;
- R5 and R6 are each independently C1-6alkyl, —C1-4-alkylene-O—Rz, —C1-4-alkylene-O—C(O)—Rz, or —C1-4-alkylene-O—C(O)—O—Rz;
- Rx and Ry are each independently C1-4alkylene;
- and
- Rz, at each occurrence, is independently C1-6alkyl, C3-10cycloalkyl, C6-20aryl, or a 4 to 8-membered heterocycle, wherein the C3-10cycloalkyl, C6-20aryl, and 4 to 8-membered heterocycle are optionally substituted with one or more substituents independently selected from the group consisting of halogen, C1-4haloalkyl, —CN, C1-4alkyl-O—, C1-4alkyl-C(O)—NH—, and C1-4alkyl-NH—C(O)—.
A method of detecting flux of a metal ion into a cell, the method comprising
- (a) contacting the cell with a compound of the following formula:
- or a salt thereof,
- wherein:
- Z is O;
- R1 is C1-10alkyl-C(O)—;
- —NR2R3 is —N(CH3)2, —N(CH2CH3)2, —N(CH2CH2CH3)2, —N(CH2CH2CH2CH3)2, —NH-cyclohexyl, —NH—CH2CH2CH2CH3, azetidin-1-yl,3,3-difluoroazetidin-1-yl, pyrrolidin-1-yl, or piperidin-1-yl;
- R4 is C1-10alkyl;
- R5 and R6 are each independently C1-6alkyl, —C1-4-alkylene-O—Rz, —C1-4-alkylene-O—C(O)—Rz, or —C1-4-alkylene-O—C(O)—O—Rz;
- and
- Rz, at each occurrence, is independently C1-6alkyl, C3-10cycloalkyl, C6-20aryl, or a 4 to 8-membered heterocycle, wherein the C3-10cycloalkyl, C6-20aryl, and 4 to 8-membered heterocycle are optionally substituted with one or more substituents independently selected from the group consisting of halogen, C1-4haloalkyl, —CN, C1-4alkyl-O—, C1-4alkyl-C(O)—NH—, and C1-4alkyl-NH—C(O)—;
- (b) subsequently contacting the cell with the metal ion;
- and
- (c) subsequently measuring fluorescence produced by the cell.
Dependencies are inferred from the claim text.