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US 5,405,975 AGranted
Fluorescent ion-selective diaryldiaza crown ether conjugates
AssigneeMolecular Probes, Inc.
InventorsMichael A. Kuhn, Richard P. Haugland
Life of this patent
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Abstract
This invention describes novel sensors for ions that are based on the combination of xanthylium-based dyes with metal-binding N,N'-diaryldiaza crown ethers. These sensors are primarily useful for detection and quantitation of alkali-metal ions in aqueous solution. Binding of the ion results in a change in the fluorescence properties of the indicating dye that can be correlated with the ion concentration. Methods are provided for attaching reactive groups on these sensors for conjugation to dyes, lipids and polymers and for enhancing entry of the indicators into living cells.
Claims16
A fluorescent ion-selective compound having the formula:
- ##STR5## where Y.sub.P is --(CH.sub.2 CH.sub.2 --O).sub.j --CH.sub.2 CH.sub.2 -- and Y.sub.Q is --(CH.sub.2 CH.sub.2 --O).sub.k --CH.sub.2 CH.sub.2 --;
- j and k are independently 1 or 2;
- where A is a substituted aryl of the formula:
- ##STR6## where at least one of R.sup.p, R.sup.q, R.sup.r, R.sup.s and R.sup.t is a first fluorophore according to the formula:
- (R.sup.a).sub.n (R.sup.b).sub.n' --FLUOR, wherein n=0 or 1;
- n'=0 or 1;
- R.sup.a is --OCH.sub.2 R.sup.3 --, --OR.sup.3' --, --SR.sup.3 --, --SR.sup.3' --, --NH(C.dbd.O)CH.sub.2 R.sup.3 --, --NH(C.dbd.O)R.sup.3' --, --(C.dbd.O)NHCH.sub.2 R.sup.3 --, --(C.dbd.O)NHR.sup.3' --, --NHSO.sub.2 R.sup.3 --, --NHSO.sub.2 R.sup.3' --, --NH(C.dbd.O)NHCH.sub.2 R.sup.3 --, --NH(C.dbd.O)NHR.sup.3' --, --NH(C.dbd.S)NHCH.sub.2 R.sup.3 --, or --NH(C.dbd.S)NHR.sup.3' --, where R.sup.3 is (CH.sub.2).sub.m and m=1-6, and R.sup.3' is phenylene (--C.sub.6 H.sub.4 --), carboxyphenylene (--C.sub.6 H.sub.3 COOH--) or sulfophenylene (--C.sub.6 H.sub.3 SO.sub.3 H--) or the pharmaceutically acceptable salt or ester thereof;
- R.sup.b is --NH--, --NH(C.dbd. O)--, --NH(C.dbd.S)--, --S--, --O--, --(C.dbd.O)--;
- or --CH.sub.2 -- or --(C.dbd.O)CH.sub.2 --;
- and
- --FLUOR is a pyronine, xanthene, fluorescein, rhodamine rosamine, rhodol, benzofluorescein, dibenzofluorescein, seminaphthofluorescein, or naphthofluorescein, or a pharmaceutically acceptable salt or ester thereof;
- where the remaining A substituents R.sup.p, R.sup.q, R.sup.r, R.sup.s, and R.sup.t, which may be the same or different, are independently H, CH.sub.3, NO.sub.2, CF.sub.3, F, Cl, Br, I, --OR.sup.5, --(C.dbd.O)OR.sup.5, or --OCH.sub.2 (C.dbd.O)OR.sup.5, where R.sup.5 is an alkyl group having 1-6 carbons, a benzyl (C.sub.6 H.sub.5 CH.sub.2 --), an acetoxymethyl ester or a pharmaceutically acceptable esterifying group, or a pharmaceutically acceptable salt;
- where B is a substituted aryl of the formula:
- ##STR7## where at least one of R.sup.p', R.sup.q', R.sup.r', R.sup.s' and R.sup.t' has the formula:
- (R.sup.a').sub.n (R.sup.b').sub.n' --X wherein n=0 or 1;
- n'=0 or 1;
- R.sup.a' is --OCH.sub.2 R.sup.3 --, --OR.sup.3' --, --SR.sup.3 --, --SR.sup.3' --, --NH(C.dbd.O)CH.sub.2 R.sup.3 --, --NH(C.dbd.O)R.sup.3' --, --(C.dbd.O)NHCH.sub.2 R.sup.3 --, --(C.dbd.O)NHR.sup.3', --NHSO.sub.2 R.sup.3 --, --NHSO.sub.2 R.sup.3' --, --NH(C.dbd.O)NHCH.sub.2 R.sup.3 --, --NH(C.dbd.O)NHR.sup.3' --, --NH(C.dbd.S)NHCH.sub.2 R.sup.3 --, or --NH(C.dbd.S)NHR.sup.3' --, where R.sup.3 is (CH.sub.2).sub.m and m=1-6, and R.sup.3' is phenylene (--C.sub.6 H.sub.4 --), carboxyphenylene (--C.sub.6 H.sub.3 COOH--) or sulfophenylene (--C.sub.6 H.sub.3 SO.sub.3 H--) or the pharmaceutically acceptable salt or ester thereof;
- R.sup.b' is --NH-- , --NH(C.dbd.O)--, --NH(C.dbd.S), --S--, --O--, --(C.dbd.O)';
- or --CH.sub.2 -- or --(C.dbd.O)CH.sub.2 --;
- and
- --X is second fluorophore --FLUOR' that is a pyronine, xanthene, fluorescein, rhodamine, rosamine, rhodol, benzofluorescein, dibenzofluorescein, seminaphthofluorescein, or naphthofluorescein, or a pharmaceutically acceptable salt or ester thereof;
- where the remaining B substituents R.sup.p', R.sup.q', R.sup.r', R.sup.s' or R.sup.t', which may be the same or different, are independently H, CH.sub.3, NO.sub.2, CF.sub.3, F, Cl, Br, I, --OR.sup.5, --(C.dbd.O)OR.sup.5, or --OCH.sub.2 (C.dbd.O)OR.sup.5 , where R.sup.5 is an alkyl group having 1-6 carbons, a benzyl (C.sub.6 H.sub.5 CH.sub.2 --), an acetoxymethyl ester or a pharmaceutically acceptable esterifying group, or a pharmaceutically acceptable salt.
19 more elements
A fluorescent ion-selective compound having the formula:
- ##STR8## where Y.sub.P is --(CH.sub.2 CH.sub.2 --O).sub.j --CH.sub.2 CH.sub.2 -- and Y.sub.Q is --(CH.sub.2 CH.sub.2 --O).sub.k --CH.sub.2 CH.sub.2 --;
- j and k are independently 1 or 2;
- --FLUOR is a pyronine, xanthene, fluorescein, rhodamine, rosamine, rhodol, benzofluorescein, dibenzofluorescein, seminaphthofluorescein, or naphthofluorescein, or a pharmaceutically acceptable salt or ester thereof that is linked to substituted aryl A by a first linkage that is --(R.sup.a).sub.n (R.sup.b).sub.n' --, wherein n=0 or 1;
- n'=0 or 1;
- R.sup.a is --OCH.sub.2 R.sup.3 --, --OR.sup.3' --, --SR.sup.3 --, --SR.sup.3' --, --NH(C.dbd.O)CH.sub.2 R.sup.3 --, --NH(C.dbd.O)R.sup.3' --, --(C.dbd.O)NHCH.sub.2 R.sup.3 --, --(C.dbd.O)NHR.sup.3' --, --NHSO.sub.2 R.sup.3 --, --NHSO.sub.2 R.sup.3' --, --NH(C.dbd.O)NHCH.sub.2 R.sup.3 --, --NH(C.dbd.O)NHR.sup.3' --, --NH(C.dbd.S)NHCH.sub.2 R.sup.3 --, or --NH(C.dbd.S)NHR.sup.3' --, where R.sup.3 is (CH.sub.2).sub.m and m=1-6, and R.sup.3' is phenylene (--C.sub.6 H.sub.4 -- ), carboxyphenylene (--C.sub.6 H.sub.3 COOH--) or sulfophenylene (--C.sub.6 H.sub.3 SO.sub.3 H--) or the pharmaceutically acceptable salt or ester thereof;
- R.sup.b is --NH--, --NH(C.dbd.O)--, --NH(C.dbd.S)--, --S--, --O--, --(C.dbd.O)--;
- or --CH.sub.2 -- or --(C.dbd.O)CH.sub.2 --;
- and
- --FLUOR', which is the same as or different from --FLUOR, is a pyronine, xanthene, fluorescein, rhodamine, rosamine, rhodol, benzofluorescein, dibenzofluorescein, seminaphthofluorescein, or naphthofluorescein, or a pharmaceutically acceptable salt or ester thereof that is linked to substituted aryl B by a second linkage that is --(R.sup.a').sub.n (R.sup.b').sub.n' --, wherein n=0 or 1;
- n'=0 or 1;
- R.sup.a' is --OCH.sub.2 R.sup.3 --, --OR.sup.3' --, --SR.sup.3 --, --SR.sup.3' --, --NH(C.dbd.O)CH.sub.2 R.sup.3 --, --NH(C.dbd.O)R.sup.3' --, --(C.dbd.O)NHCH.sub.2 R.sup.3 --, --(C.dbd.O)NHR.sup.3', --NHSO.sub.2 R.sup.3 --, --NHSO.sub.2 R.sup.3' --, --NH(C.dbd.O)NHCH.sub.2 R.sup.3 --, --NH(C.dbd.O)NHR.sup.3' --, --NH(C.dbd.S)NHCH.sub.2 R.sup.3 --, or --NH(C.dbd.S)NHR.sup.3' --, where R.sup.3 is (CH.sub.2).sub.m and m=1-6, and R.sup.3' is phenylene (--C.sub.6 H.sub.4 --), carboxyphenylene (--C.sub.6 H.sub.3 COOH--) or sulfophenylene (--C.sub.6 H.sub.3 SO.sub.3 H--) or the pharmaceutically acceptable salt or ester thereof;
- R.sup.b' is --NH--, --NH(C.dbd.O)--, --NH(C.dbd.S), --S--, --O--, --(C.dbd.O)--;
- or --CH.sub.2 -- or --(C.dbd.O)CH.sub.2 --;
- and
- the remaining A and B substituents, which may be the same or different, are independently H, CH.sub.3, NO.sub.2, CF.sub.3, F, Cl, Br, I, --OR.sup.5, --(C.dbd.O)OR.sup.5, or --OCH.sub.2 (C.dbd.O)OR.sup.5, where R.sup.5 is an alkyl group having 1-6 carbons, a benzyl (C.sub.6 H.sub.5 CH.sub.2 --), an acetoxymethyl ester or a pharmaceutically acceptable esterifying group, or a pharmaceutically acceptable salt.
12 more elements
A fluorescent ion-selective compound comprising N,N'-diaryldiazacrown ether that is a DA15C5 or DA18C6 containing two aryl residues, wherein one aryl residue attached to the crown ether is covalently linked to a pyronine, xanthene, fluorescein, rhodamine, rosamine, rhodol, benzofluorescein, dibenzofluorescein, seminaphthofluorescein, or naphthofluorescein, or a pharmaceutically acceptable salt or ester thereof, and the other aryl residue attached to the crown ether is covalently linked to an additional pyronine, xanthene, fluorescein, rhodamine, rosamine, rhodol, benzofluorescein, dibenzofluorescein, seminaphthofluorescein, or naphthofluorescein, or a pharmaceutically acceptable salt or ester thereof.
Dependencies are inferred from the claim text.