Substituted unsymmetrical cyanine dyes with selected permeability
First Claim
Patent Images
1. A compound of the formula ##STR109## wherein each R1 is independently H;
- or an alkyl group having from 1-6 carbons;
an alkoxy group having from 1-6 carbons;
or a trifluoromethyl;
or a halogen; and
t=1-4;
R2 is an alkyl group having 1-6 carbons;
X is O, S, Se or NR15, where R15 is H or an alkyl group having 1-6 carbons;
or X is CR16 R17 where R16 and R17, which may be the same or different, are independently alkyl groups having 1-6 carbons, or R16 and R17 taken in combination complete a five or six membered saturated ring;
n=0, 1 or2;
Ψ
- is a biologically compatible counterion;
Q has the formula Q1 or Q2 ##STR110## wherein Y is --CR3 ═
CR4 --;
p and m=0 or 1, such that p+m=1;
R5 is an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or R5 is a cyclic substituent that is a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or R5 is a TAIL;
R3, R4, R6 and R7, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;
where R8 and R9, which can be the same or different, are independently H;
or alkyl groups having 1-6 carbons;
or 1-2 alicyclic or aromatic rings;
or R8 and R9 taken in combination are --(CH2)4 -- or --(CH2)5 -- to give a 5 or 6 membered ring; and
where R19 is alkyl having 1-6 carbons, or perfluoroalkyl having 1-6 carbons, or aryl;
or R6 and R7, taken in combination are --(CH2)v -- where v=3 or 4, or R6 and R7 form a fused aromatic ring according to formula Q2;
R11, R12, R13, and R14, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a TAIL;
or --OH, --OR8, --SR8, or --(NR8 R9);
TAIL is a heteroatom-containing moiety having the formula LINK-SPACER-CAP;
whereinLINK is a single covalent bond, --O--, --S--, or --NR20 --;
where R20 is H, a linear or branched alkyl having 1-8 carbons, or R20 is --SPACER'"'"'--CAP'"'"';
SPACER and SPACER'"'"', which may be the same or different, are covalent linkages, linear or branched, cyclic or heterocyclic, saturated or unsaturated, each having 1-16 nonhydrogen atoms selected from the group consisting of C, N, P, O and S, such that the linkage contains any combination of ether, thioether, amine, ester, amide bonds;
or single, double, triple or aromatic carbon-carbon bonds;
or phosphorus-oxygen, phosphorus-sulfur bonds, nitrogen-nitrogen or nitrogen-oxygen bonds;
or aromatic or heteroaromatic bonds;
CAP and CAP'"'"', which may be the same or different, are --O--R21, --S--R21, --NR21 R22, or --N+ R21 R22 R23 Ψ
- ;
whereinR21, R22, and R23 are independently H, or a linear or branched alkyl or cycloalkyl having 1-8 carbons, optionally further substituted by hydroxy, alkoxy having 1-8 carbons, carboxyalkyl having 1-8 carbons, or phenyl, where phenyl is optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, aminoalkyl having 1-8 carbons, or carboxyalkyl having 1-8 carbons;
or, one or more of R21, R22 and R23, taken in combination with SPACER or SPACER'"'"' or R20 forms a 5- or 6-membered aromatic, heteroaromatic, alicyclic or heteroalicyclic ring, the heteroatoms selected from O, N or S;
where Ψ
- is a biologically compatible counterion;
orCAP and CAP'"'"' are independently ##STR111## where R21, R22 and Ψ
- are as defined previously;
such that at least one of R3, R4, R5, R6, R7, R11, R12, R13, and R14 is a TAIL; and
when R5 is a TAIL, LINK is a single bond; and
where only R5 is a TAIL, R3 or R4 is not hydrogen; and
, where more than one of R2, R3, R4, R5, R6, R7, R11, R12, R13, and R14 is a TAIL, each TAIL is optionally the same or different.
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Abstract
The invention describes the preparation and use of fluorescent stains for nucleic acids derived from unsymmetrical cyanine dyes comprising a substituted benzazolium ring system linked by a methine bridge to a pyridinium or quinolinium ring system having at least one substituent on the pyridinium or quinolinium ring that contains a heteroatom. The presence of the heteroatom-containing substituent results in higher sensitivity to oligonucleotides and larger nucleic acid polymers in a wide range of cells and gels, and for use in analysis of cell structure, membrane integrity or function.
357 Citations
97 Claims
-
1. A compound of the formula ##STR109## wherein each R1 is independently H;
- or an alkyl group having from 1-6 carbons;
an alkoxy group having from 1-6 carbons;
or a trifluoromethyl;
or a halogen; and
t=1-4;R2 is an alkyl group having 1-6 carbons; X is O, S, Se or NR15, where R15 is H or an alkyl group having 1-6 carbons;
or X is CR16 R17 where R16 and R17, which may be the same or different, are independently alkyl groups having 1-6 carbons, or R16 and R17 taken in combination complete a five or six membered saturated ring;n=0, 1 or2; Ψ
- is a biologically compatible counterion;Q has the formula Q1 or Q2 ##STR110## wherein Y is --CR3 ═
CR4 --;p and m=0 or 1, such that p+m=1; R5 is an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or R5 is a cyclic substituent that is a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or R5 is a TAIL;R3, R4, R6 and R7, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;
where R8 and R9, which can be the same or different, are independently H;
or alkyl groups having 1-6 carbons;
or 1-2 alicyclic or aromatic rings;
or R8 and R9 taken in combination are --(CH2)4 -- or --(CH2)5 -- to give a 5 or 6 membered ring; and
where R19 is alkyl having 1-6 carbons, or perfluoroalkyl having 1-6 carbons, or aryl;or R6 and R7, taken in combination are --(CH2)v -- where v=3 or 4, or R6 and R7 form a fused aromatic ring according to formula Q2; R11, R12, R13, and R14, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a TAIL;
or --OH, --OR8, --SR8, or --(NR8 R9);TAIL is a heteroatom-containing moiety having the formula LINK-SPACER-CAP; wherein LINK is a single covalent bond, --O--, --S--, or --NR20 --;
where R20 is H, a linear or branched alkyl having 1-8 carbons, or R20 is --SPACER'"'"'--CAP'"'"';SPACER and SPACER'"'"', which may be the same or different, are covalent linkages, linear or branched, cyclic or heterocyclic, saturated or unsaturated, each having 1-16 nonhydrogen atoms selected from the group consisting of C, N, P, O and S, such that the linkage contains any combination of ether, thioether, amine, ester, amide bonds;
or single, double, triple or aromatic carbon-carbon bonds;
or phosphorus-oxygen, phosphorus-sulfur bonds, nitrogen-nitrogen or nitrogen-oxygen bonds;
or aromatic or heteroaromatic bonds;CAP and CAP'"'"', which may be the same or different, are --O--R21, --S--R21, --NR21 R22, or --N+ R21 R22 R23 Ψ
- ;wherein R21, R22, and R23 are independently H, or a linear or branched alkyl or cycloalkyl having 1-8 carbons, optionally further substituted by hydroxy, alkoxy having 1-8 carbons, carboxyalkyl having 1-8 carbons, or phenyl, where phenyl is optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, aminoalkyl having 1-8 carbons, or carboxyalkyl having 1-8 carbons;
or, one or more of R21, R22 and R23, taken in combination with SPACER or SPACER'"'"' or R20 forms a 5- or 6-membered aromatic, heteroaromatic, alicyclic or heteroalicyclic ring, the heteroatoms selected from O, N or S;
where Ψ
- is a biologically compatible counterion;or CAP and CAP'"'"' are independently ##STR111## where R21, R22 and Ψ
- are as defined previously;
such that at least one of R3, R4, R5, R6, R7, R11, R12, R13, and R14 is a TAIL; and
when R5 is a TAIL, LINK is a single bond; and
where only R5 is a TAIL, R3 or R4 is not hydrogen; and
, where more than one of R2, R3, R4, R5, R6, R7, R11, R12, R13, and R14 is a TAIL, each TAIL is optionally the same or different. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
- or an alkyl group having from 1-6 carbons;
-
19. A compound of the formula ##STR114## or having the formula ##STR115## wherein R2 is an alkyl group having 1-6 carbons;
-
X is O or S; n=0, 1 or2; Ψ
- is a biologically compatible counterion;R5 is a substituted or unsubstituted naphthyl, phenyl, thienyl, or cycloalkyl having 3-8 carbons; R4 is an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8,--(NR8 R9);
or --OSO2 R19 ;
or a TAIL;
where R8 and R9, which can be the same or different, are independently H;
or alkyl groups having 1-6 carbons;
or 1-2 alicyclic or aromatic rings;
or R8 and R9 taken in combination are --(CH2)4 -- or --(CH2)5 -- to give a 5 or 6 membered ring; and
where R19 is alkyl having 1-6 carbons, or perfluoroalkyl having 1-6 carbons, or aryl;
having 1-6 carbons, or aryl;R3, R6 and R7, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;R11, R12, R13, and R14, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-8 carbons;
or a TAIL;
or --OR8, --SR8, or --(NR8 R9);TAIL is a heteroatom-containing moiety having the formula LINK-SPACER-CAP; wherein LINK is a single covalent bond, --O--, --S--, or --NR20 --;
where R20 is H, a linear or branched alkyl having 1-8 carbons, or R20 is --SPACER'"'"'--CAP'"'"';SPACER and SPACER'"'"', which may be the same or different, are covalent linkages, linear or branched, cyclic or heterocyclic, saturated or unsaturated, each having 1-16 nonhydrogen atoms selected from the group consisting of C, N, O and S, such that the linkage begins and ends with a carbon atom, and contains any combination of ether, thioether, amine, ester, amide, or aliphatic, oleic or aromatic carbon-carbon bonds, or aromatic carbon-nitrogen or nitrogen-nitrogen bonds;
wherein all heteroatoms in the linear backbone of SPACER are separated by at least two carbon atoms;CAP and CAP'"'"', which may be the same or different, are --O--R21, --S--R21, --NR21 R22, or --N+ R21 R22 R23 Ψ
- ;wherein R21, R22, and R23 are independently H, or a linear or branched alkyl or cycloalkyl having 1-8 carbons, optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, carboxyalkyl having 1-8 carbons, or phenyl, where phenyl is optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, aminoalkyl having 1-8 carbons, or carboxyalkyl having 1-8 carbons;
or, one or more of R21, R22 and R23, taken in combination with SPACER or SPACER'"'"' or R20 forms a 5- or 6-membered aromatic, heteroaromatic, alicyclic or heteroalicyclic ring, the heteroatoms selected from O, N or S;
where Ψ
- is a biologically compatible counterion;or CAP and CAP'"'"' are independently ##STR116## where R21, R22 and Ψ
- are as defined previously;
such that at least one of R3, R4, R6, R7, R11, R12, R13, and R14 is a TAIL; and
, where more than one substituent is a TAIL, each TAIL is optionally the same or different. - View Dependent Claims (20)
-
-
21. A compound having the formula ##STR117## or having the formula ##STR118## wherein R2 is an alkyl group having 1-6 carbons;
-
X is O or S; n=0, 1 or2; Ψ
- is a biologically compatible counterion;R5 is an alkyl having 1-6 carbons; R4 is an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR.sup. R9);
or --OSO2 R19 ;
or a TAIL;
where R8 and R9, which can be the same or different, are independently H;
or alkyl groups having 1-6 carbons;
or 1-2 alicyclic or aromatic rings;
or R8 and R9 taken in combination are --(CH2)4 -- or --(CH2)5 -- to give a 5 or 6 membered ring; and
where R19 is alkyl having 1-6 carbons, or perfluoroalkyl having 1-6 carbons, or aryl;
having 1-6 carbons, or aryl;R3, R6 and R7, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;R11, R12, R13, and R14, which may be the same or different, are independently H;
or an alkyl, alkenyl polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-8 carbons;
or a TAIL;
or --OR8, --SR8, or --(NR8 R9);TAIL is a heteroatom-containing moiety having the formula LINK-SPACER-CAP; wherein LINK is a single covalent bond, --O--, --S--, or --NR20 --;
where R20 is H, a linear or branched alkyl having 1-8 carbons, or R20 is --SPACER'"'"'--CAP'"'"';SPACER and SPACER'"'"', which may be the same or different, are covalent linkages, linear or branched, cyclic or heterocyclic, saturated or unsaturated, each having 1-16 nonhydrogen atoms selected from the group consisting of C, N, O and S, such that the linkage begins and ends with a carbon atom, and contains any combination of ether, thioether, amine, ester, amide, or aliphatic, olefinic or aromatic carbon-carbon bonds, or aromatic carbon-nitrogen or nitrogen-nitrogen bonds;
wherein all heteroatoms in the linear backbone of SPACER are separated by at least two carbon atoms;CAP and CAP'"'"', which may be the same or different, are --O--R21, --S--R21, --NR21 R22, or --N+ R21 R23 Ψ
- ;wherein R21, R22, and R23 are independently H, or a linear or branched alkyl or cycloalkyl having 1-8 carbons, optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, carboxyalkyl having 1-8 carbons, or phenyl, where phenyl is optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, aminoalkyl having 1-8 carbons, or carboxyalkyl having 1-8 carbons;
or, one or more of R21, R22 and R23, taken in combination with SPACER or SPACER'"'"' or R20 forms a 5- or 6-membered aromatic, heteroaromatic, alicyclic or heteroalicyclic ring, the heteroatoms selected from O, N or S;
where Ψ
- is a biologically compatible counterion;or CAP and CAP'"'"' are independently ##STR119## where R21, R22 and Ψ
- are as defined previously;
such that at least one of R3, R4, R6, R7, R11, R12, R13, and R14 is a TAIL; and
, where more than one substituent is a TAIL, each TAIL is optionally the same or different.
-
-
22. A compound having the formula ##STR120## wherein R2 and R24 are independently a linear or branched alkyl having 1-6 carbons;
-
X is O, S, Se or NR15, where R15 is H or an alkyl group having 1-6 carbons;
or X is C(CH3)2 ;n=0, 1 or2; and Ψ
- is a biologically compatible counterion.
-
-
23. A fluorescent complex comprising a nucleic acid polymer non-covalently bound to one or more dye molecules of the formula ##STR121## wherein each R1 is independently H;
- or an alkyl group having from 1-6 carbons;
an alkoxy group having from 1-6 carbons;
or a trifluoromethyl;
or a halogen; and
t=1-4;R2 is an alkyl group having 1-6 carbons; X is O, S, Se or NR15, where R15 is H or an alkyl group having 1-6 carbons;
or X is CR16 R17 where R16 and R17, which may be the same or different, are independently alkyl groups having 1-6 carbons, or R16 and R17 taken in combination complete a five or six membered saturated ring;n=0, 1 or 2; Ψ
- is a biologically compatible counterion;Q has the formula Q 1 or Q2 ##STR122## wherein Y is --CR3 ═
CR4 --;p and m=0 or 1, such that p+m=1; R5 is an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or R5 is a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or R5 is a TAIL;R3, R4, R6 and R7, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;
where R8 and R9, which can be the same or different, are independently H;
or alkyl groups having 1-6 carbons;
or 1-2 alicyclic or aromatic rings;
or R8 and R9 taken in combination are --(CH2)4 -- or --(CH2)5 -- to give a 5 or 6 membered ring; and
where R19 is alkyl having 1-6 carbons, or perfluoroalkyl having 1-6 carbons, or aryl;or R6 and R7, taken in combination are --(CH2)v -- where v=3 or 4, or R6 and R7 form a fused aromatic ring according to formula Q2; R11, R12, R13, and R14, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a TAIL;
or --OH, --OR8, --SR8, or --(NR8 R9);TAIL is a heteroatom-containing moiety having the formula LINK-SPACER-CAP; wherein LINK is a single covalent bond, --O--, --S--, or --NR20 --;
where R20 is H, a linear or branched alkyl having 1-8 carbons, or R20 is --SPACER'"'"'--CAP'"'"';SPACER and SPACER'"'"', which may be the same or different, are covalent linkages, linear or branched, cyclic or heterocyclic, saturated or unsaturated, each having 1-16 nonhydrogen atoms selected from the group consisting of C, N, P, O and S, such that the linkage contains any combination of ether, thioether, amine, ester, amide bonds;
or single, double, triple or aromatic carbon-carbon bonds;
or phosphorus-oxygen, phosphorus-sulfur bonds, nitrogen-nitrogen or nitrogen-oxygen bonds;
or aromatic or heteroaromatic bonds;CAP and CAP'"'"', which may be the same or different, are --O--R21, --S--R21, --NR21 R22, or --N+ R21 R22 R23 Ψ
- ;wherein R21, R22, and R23 are independently H, or a linear or branched alkyl or cycloalkyl having 1-8 carbons, optionally further substituted by hydroxy, alkoxy having 1-8 carbons, carboxyalkyl having 1-8 carbons, or phenyl, where phenyl is optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, aminoalkyl having 1-8 carbons, or carboxyalkyl having 1-8 carbons;
or, one or more of R21, R22 and R23, taken in combination with SPACER or SPACER'"'"' or R20 forms a 5- or 6-membered aromatic, heteroaromatic, alicyclic or heteroalicyclic ring, the heteroatoms selected from O, N or S;
where Ψ
- is a biologically compatible counterion;CAP and CAP'"'"' are independently ##STR123## where R21, R22 and Ψ
- are as defined previously;
such that at least one of R3, R4, R5, R6, R7, R11, R12, R13, and R14 is a TAIL, and when R5 is a TAIL, LINK is a single bond; and
where only R5 is a TAIL, R3 or R4 is not hydrogen; and
, where more than one of R2, R3, R4, R5, R6, R7, R11, R12, R13, and R14 is a TAIL, each TAIL is optionally the same or different. - View Dependent Claims (24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34)
- or an alkyl group having from 1-6 carbons;
-
35. A method of staining nucleic acids, comprising
a) combining a sample that contains or is thought to contain a nucleic acid, with a mixture containing a dye compound of the formula (formula I): - ##STR124## or having the formula ##STR125## wherein R2 is ethyl or methyl;
X is O or S; n=0, 1 or2; Ψ
- is a biologically compatible counterion;R5 is alkyl having 1-6 carbons;
or a cyclic substituent that is a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or R5 is a TAIL;R4 is an alkyl, aIkenyl, polyaIkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;
where R8 and R9, which can be the same or different, are independently H;
or alkyl groups having 1-6 carbons;
or 1-2 alicyclic or aromatic rings;
or R8 and R9 taken in combination are --(CH2)4 -- or --(CH2)5 -- to give a 5 or 6 membered ring; and
where R19 is alkyl having 1-6 carbons, or perfluoroalkyl having 1-6 carbons, or aryl;
having 1-6 carbons, or aryl;R3, R6 and R7, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;R11, R12, R13, and R14, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-8 carbons;
or a TAIL;
or --OR8, --SR8, or --(NR8 R9);TAIL is a heteroatom-containing moiety having the formula LINK-SPACER-CAP; wherein LINK is a single covalent bond, --O--, --S--, or --NR20 --;
where R20 is H, a linear or branched alkyl having 1-8 carbons, or R20 is --SPACER'"'"'--CAP'"'"';SPACER and SPACER'"'"', which may be the same or different, are covalent linkages, linear or branched, cyclic or heterocyclic, saturated or unsaturated, each having 1-16 nonhydrogen atoms selected from the group consisting of C, N, O and S, such that the linkage begins and ends with a carbon atom, and contains any combination of ether, thioether, amine, ester, amide, or aliphatic, olefinic or aromatic carbon-carbon bonds, or aromatic carbon-nitrogen or nitrogen-nitrogen bonds;
wherein all heteroatoms in the linear backbone of SPACER are separated by at least two carbon atoms;CAP and CAP'"'"', which may be the same or different, are --O--R21, --S--R21, --NR21 R22, or --N+ R21 R22 R23 Ψ
- ;wherein R21, R22, and R23 are independently H, or a linear or branched alkyl or cycloalkyl having 1-8 carbons, optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, carboxyalkyl having 1-8 carbons, or phenyl, where phenyl is optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, aminoalkyl having 1-8 carbons, or carboxyalkyl having 1-8 carbons;
or, one or more of R21, R22 and R23, taken in combination with SPACER or SPACER'"'"' or R20 forms a 5- or 6-membered aromatic, heteroaromatic, alicyclic or heteroalicyclic ring, the heteroatoms selected from O, N or S;
where Ψ
- is a biologically compatible counterion;or CAP and CAP'"'"' are independently ##STR126## where R21, R22 and Ψ
- are as defined previously;
such that at least one of R3, R4, R6, R7, R11, R12, R13, and R14 is a TAIL; and
, where more than one substituent is a TAIL, each TAIL is optionally the same or different;and b) incubating the sample and mixture for a time sufficient for the dye compound to combine with the nucleic acid in the sample to form one or more dye-nucleic acid complexes that give a detectable fluorescent signal. - View Dependent Claims (36, 37, 38, 39, 40, 41, 42)
- ##STR124## or having the formula ##STR125## wherein R2 is ethyl or methyl;
-
43. A method of detecting a biological structure, comprising
a) combining a sample that contains or is thought to contain a specific biological structure, with a dye of the formula ##STR127## or having the formula ##STR128## wherein R2 is ethyl or methyl; -
X is O or S; n=0, 1 or2; Ψ
- is a biologically compatible counterion;R5 is alkyl having 1-6 carbons;
or a cyclic substituent that is a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or R5 is a TAIL;R4 is an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, (NR8 R9);
or --OSO2 R19 ;
or a TAIL;
where R8 and R9, which can be the same or different, are independently H;
or alkyl groups having 1-6 carbons;
or 1-2 alicyclic or aromatic rings;
or R8 and R9 taken in combination are --(CH2)4 -- or --(CH2)5 -- to give a 5 or 6 membered ring; and
where R19 is alkyl having 1-6 carbons, or perfluoroalkyl having 1-6 carbons, or aryl;
having 1-6 carbons, or aryl;R3, R6 and R7, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;R11, R12, R13, and R14, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-8 carbons;
or a TAIL;
or --OR8, --SR8, or --(NR8 R9);TAIL is a heteroatom-containing moiety having the formula LINK-SPACER-CAP; wherein LINK is a single covalent bond, --O--, --S--, or --NR20 --;
where R20 is H, a linear or branched alkyl having 1-8 carbons, or R20 is --SPACER'"'"'--CAP'"'"';SPACER and SPACER'"'"', which may be the same or different, are covalent linkages, linear or branched, cyclic or heterocyclic, saturated or unsaturated, each having 1-16 nonhydrogen atoms selected from the group consisting of C, N, O and S, such that the linkage begins and ends with a carbon atom, and contains any combination of ether, thioether, amine, ester, amide, or aliphatic, olefinic or aromatic carbon-carbon bonds, or aromatic carbon-nitrogen or nitrogen-nitrogen bonds;
wherein all heteroatoms in the linear backbone of SPACER are separated by at least two carbon atoms;CAP and CAP'"'"', which may be the same or different, are --O--R21, --S--R21, --NR21 R22, or --N+ R21 R22 R23 Ψ
-,wherein R21, R22, and R23 are independently H, or a linear or branched alkyl or cycloalkyl having 1-8 carbons, optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, carboxyalkyl having 1-8 carbons, or phenyl, where phenyl is optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, aminoalkyl having 1-8 carbons, or carboxyalkyl having 1-8 carbons;
or, one or more of R21, R22 and R23, taken in combination with SPACER or SPACER'"'"' or R20 forms a 5- or 6-membered aromatic, heteroaromatic, alicyclic or heteroalicyclic ring, the heteroatoms selected from O, N or S;
where Ψ
- is a biologically compatible counterion;or CAP and CAP'"'"' are independently ##STR129## where R21, R22 and Ψ
- are as defined previously;
such that at least one of R3, R4, R6, R7, R11, R12, R13, and R14 is a TAIL; and
, where more than one substituent is a TAIL, each TAIL is optionally the same or different;wherein said biological structure contains nucleic acids; b) incubating the combined sample and dye compound for a time sufficient for the dye compound to combine with the nucleic acids in the biological structure to form a pattern of dye-nucleic acid complexes having a detectable fluorescent signal that corresponds to the biological structure; and c) detecting the fluorescent signal that corresponds to the biological structure. - View Dependent Claims (44, 45, 46, 47, 48, 49)
-
-
50. A method of characterizing a sample, comprising:
-
a) combining a sample containing or thought to contain nucleic acid polymers with a dye compound of the formula ##STR130## or having the formula ##STR131## wherein R2 is ethyl or methyl; X is O or S; n=0, 1 or2; Ψ
- is a biologically compatible counterion;R5 is alkyl having 1-6 carbons;
or a cyclic substituent that is a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or R5 is a TAIL;R4 is an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;
where R8 and R9, which can be the same or different, are independently H;
or alkyl groups having 1-6 carbons;
or 1-2 alicyclic or aromatic rings;
or R8 and R9 taken in combination are --(CH2)4 -- or --(CH2)5 -- to give a 5 or 6 membered ring; and
where R19 is alkyl having 1-6 carbons, or perfluoroalkyl having 1-6 carbons, or aryl;
having 1-6 carbons, or aryl;R3, R6 and R7, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;R11, R12, R13, and R14, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-8 carbons;
or a TAIL;
or --OR8, --SR8, or --(NR8 R9);TAIL is a heteroatom-containing moiety having the formula LINK-SPACER-CAP; wherein LINK is a single covalent bond, --O--, --S--, or --NR20 --;
where R20 is H, a linear or branched alkyl having 1-8 carbons, or R20 is --SPACER'"'"'--CAP'"'"';SPACER and SPACER'"'"', which may be the same or different, are covalent linkages, linear or branched, cyclic or heterocyclic, saturated or unsaturated, each having 1-16 nonhydrogen atoms selected from the group consisting of C, N, O and S, such that the linkage begins and ends with a carbon atom, and contains any combination of ether, thioether, amine, ester, amide, or aliphatic, olefinic or aromatic carbon-carbon bonds, or aromatic carbon-nitrogen or nitrogen-nitrogen bonds;
wherein all heteroatoms in the linear backbone of SPACER are separated by at least two carbon atoms;CAP and CAP'"'"', which may be the same or different, are --O--R21, --S--R21, --NR21 R22, or --N+ R21 R22 R23 Ψ
- ;wherein R21, R22, and R23 are independently H, or a linear or branched alkyl or cycloalkyl having 1-8 carbons, optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, carboxyalkyl having 1-8 carbons, or phenyl, where phenyl is optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, aminoalkyl having 1-8 carbons, or carboxyalkyl having 1-8 carbons;
or, one or more of R21, R22 and R23, taken in combination with SPACER or SPACER'"'"' or R20 forms a 5- or 6-membered aromatic, heteroaromatic, alicyclic or heteroalicyclic ring, the heteroatoms selected from O, N or S;
where Ψ
- is a biologically compatible counterion;or CAP and CAP'"'"' are independently ##STR132## where R21, R22 and Ψ
- are as defined previously;
such that at least one of R3, R4, R6, R7, R11, R12, R13, and R14 is a TAIL; and
, where more than one substituent is a TAIL, each TAIL is optionally the same or different;b) incubating the sample for a time sufficient for the dye compound to combine with nucleic acid polymers in the sample to form a staining profile of dye-nucleic acid complexes having detectable fluorescent signals, said staining pattern having a shape, location, spectral property, or intensity that is indicative of a characteristic of the sample; and c) characterizing the sample based on said staining profile. - View Dependent Claims (51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62)
-
-
63. A method of determining cell membrane integrity comprising:
-
a) incubating a sample containing one or more cells with a first dye compound of the formula ##STR133## or having the formula ##STR134## wherein R2 is ethyl or methyl; X is O or S; n=0, 1 or2; Ψ
- is a biologically compatible counterion;R5 is alkyl having 1-6 carbons;
or a cyclic substituent that is a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or R5 is a TAIL;R4 is an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;
where R8 and R9, which can be the same or different, are independently H;
or alkyl groups having 1-6 carbons;
or 1-2 alicyclic or aromatic rings;
or R8 and R9 taken in combination are --(CH2)4 -- or --(CH2)5 -- to give a 5 or 6 membered ring; and
where R19 is alkyl having 1-6 carbons, or perfluoroalkyl having 1-6 carbons, or aryl;
having 1-6 carbons, or aryl;R3, R6 and R7, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;R11, R12, R13, and R14, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-8 carbons;
or a TAIL;
or --OR8, --SR8, or --(NR8 R9);TAIL is a heteroatom-containing moiety having the formula LINK-SPACER-CAP; wherein LINK is single covalent bond, --O--, --S--, or --NR20 --;
where R20 is H, a linear or branched alkyl having 1-8 carbons, or R20 is --SPACER'"'"'--CAP'"'"';SPACER and SPACER'"'"', which may be the same or different, are covalent linkages, linear or branched, cyclic or heterocyclic, saturated or unsaturated, each having 1-16 nonhydrogen atoms selected from the group consisting of C, N, O and S, such that the linkage begins and ends with a carbon atom, and contains any combination of ether, thioether, amine, ester, amide, or aliphatic, olefinic or aromatic carbon-carbon bonds, or aromatic carbon-nitrogen or nitrogen-nitrogen bonds;
wherein all heteroatoms in the linear backbone of SPACER are separated by at least two carbon atoms;CAP and CAP'"'"', which may be the same or different, are --NR21 R22, or --N+ R21 R22 R23 Ψ
- ;wherein R21, R22, and R23 are independently H, or a linear or branched alkyl or cycloalkyl having 1-8 carbons, optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, carboxyalkyl having 1-8 carbons, or phenyl, where phenyl is optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, aminoalkyl having 1-8 carbons, or carboxyalkyl having 1-8 carbons;
or, one or more of R21, R22 and R23, taken in combination with SPACER or SPACER'"'"' or R20 forms a 5- or 6-membered aromatic, heteroaromatic, alicyclic or heteroalicyclic ring, the heteroatoms selected from O, N or S;
where Ψ
- is a biologically compatible counterion;or CAP and CAP'"'"' are independently ##STR135## where R21, R22 and Ψ
- are as defined previously;
such that at least one of R3, R4, R6, R7, R11, R12, R13, and R14 is a TAIL; and
, where more than one substituent is a TAIL, each TAIL is optionally the same or different;for a time sufficient for the dye compound to combine with intracellular nucleic acids to form a first intracellular dye-nucleic acid complex having a detectable fluorescent signal; and c) determining cell membrane integrity of cells in the sample based on presence of the detectable fluorescent signal, where the presence of the detectable fluorescent signal indicates that the cell membrane integrity is compromised and the absence of the detectable fluorescent signal indicates that the cell membrane integrity is intact. - View Dependent Claims (64, 65, 66, 67)
-
-
68. A method of quantitating nucleic acids in a sample, comprising
a) combining an aliquot of a sample, optionally the entire sample, that contains or is thought to contain a nucleic acid, with mixture containing a dye compound of the formula ##STR136## or having the formula ##STR137## wherein R2 is ethyl or methyl; -
X is O or S; n=0, 1 or2; Ψ
- is a biologically compatible counterion;R5 is alkyl having 1-6 carbons;
or a cyclic substituent that is a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or R5 is a TAIL;R4 is an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;
where R8 and R9, which can be the same or different, are independently H;
or alkyl groups having 1-6 carbons;
or 1-2 alicyclic or aromatic rings;
or R8 and R9 taken in combination are --(CH2)4 -- or --(CH2)5 -- to give a 5 or 6 membered ring; and
where R19 is alkyl having 1-6 carbons, or perfluoroalkyl having 1-6 carbons, or aryl;
having 1-6 carbons, or aryl;R3, R6 and R7, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;R11, R12, R13, and R14, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-8 carbons;
or a TAIL;
or --OR8, --SR8, or --(NR8 R9);TAIL is a heteroatom-containing moiety having the formula LINK-SPACER-CAP; wherein LINK is a single covalent bond, --O--, --S--, or --NR20 --;
where R20 is H, a linear or branched alkyl having 1-8 carbons, or R20 is --SPACER'"'"'--CAP'"'"';SPACER and SPACER'"'"', which may be the same or different, are covalent linkages, linear or branched, cyclic or heterocyclic, saturated or unsaturated, each having 1-16 nonhydrogen atoms selected from the group consisting of C, N, O and S, such that the linkage begins and ends with a carbon atom, and contains any combination of ether, thioether, amine, ester, amide, or aliphatic, olefinic or aromatic carbon-carbon bonds, or aromatic carbon-nitrogen or nitrogen-nitrogen bonds;
wherein all heteroatoms in the linear backbone of SPACER are separated by at least two carbon atoms;CAP and CAP'"'"', which may be the same or different, are --O--R21, --S--R21, --NR21 R22, or --N+ R21 R22 R23 Ψ
- ;wherein R21, R22, and R23 are independently H, or a linear or branched alkyl or cycloalkyl having 1-8 carbons, optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, carboxyalkyl having 1-8 carbons, or phenyl, where phenyl is optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, aminoalkyl having 1-8 carbons, or carboxyalkyl having 1-8 carbons;
or, one or more of R21, R22 and R23, taken in combination with SPACER or SPACER'"'"' or R20 forms a 5- or 6-membered aromatic, heteroaromatic, alicyclic or heteroalicyclic ring, the heteroatoms selected from O, N or S;
where Ψ
- is a biologically compatible counterion;or CAP and CAP'"'"' are independently ##STR138## where R21, R22 and Ψ
- are as defined previously;
such that at least one of R3, R4, R6, R7, R11, R12, R13, and R14 is a TAIL; and
, where more than one substituent is a TAIL, each TAIL is optionally the same or different;b) incubating the aliquot and mixture for a time sufficient for the dye compound to combine with the nucleic acid in the sample to form a dye-nucleic acid complex that gives a detectable fluorescent signal; and c) quantifying the nucleic acid present in said sample based on comparison of the detectable fluorescent signal with a fluorescent standard characteristic of a given amount of nucleic acid. - View Dependent Claims (69, 70, 71, 72, 73, 74)
-
-
75. A method of analyzing nucleic acid-analyte interactions, comprising:
-
a) forming a fluorescent complex comprising a nucleic acid polymer and one or more dye molecules of the formula;
##STR139## or having the formula ##STR140## wherein R2 is ethyl or methyl;X is O or S; n=0, 1 or2; Ψ
- is a biologically compatible counterion;R5 is alkyl having 1-6 carbons;
or a cyclic substituent that is a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or R5 is a TAIL;R4 is an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;
where R8 and R9, which can be the same or different, are independently H;
or alkyl groups having 1-6 carbons;
or 1-2 alicyclic or aromatic rings;
or R8 and R9 taken in combination are --(CH2)4 -- or --(CH2)5 -- to give a 5 or 6 membered ring, and where R19 is alkyl having 1-6 carbons, or perfluoroalkyl having 1-6 carbons, or aryl;
having 1-6 carbons, or aryl;R3, R6 and R7, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;R11, R12, R13, and R14, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-8 carbons;
or a TAIL;
or --OR8, --SR8, or --(NR8 R9);TAIL is a heteroatom-containing moiety having the formula LINK-SPACER-CAP; wherein LINK is a single covalent bond, --O--, --S--, or --NR20 --;
where R20 is H, a linear or branched alkyl having 1-8 carbons;
or R20 is --SPACER'"'"'--CAP'"'"';SPACER and SPACER'"'"', which may be the same or different, are covalent linkages, linear or branched, cyclic or heterocyclic, saturated or unsaturated, each having 1-16 nonhydrogen atoms selected from the group consisting of C, N, O and S, such that the linkage begins and ends with a carbon atom, and contains any combination of ether, thioether, amine, ester, amide, or aliphatic, olefinic or aromatic carbon-carbon bonds, or aromatic carbon-nitrogen or nitrogen-nitrogen bonds;
wherein all heteroatoms in the linear backbone of SPACER are separated by at least two carbon atoms;CAP and CAP'"'"', which may be the same or different, are --O--R21, --S--R21, --NR21 R22, or --N+ R21 R22 R23 Ψ
- ;wherein R21, R22, and R23 are independently H, or a linear or branched alkyl or cycloalkyl having 1-8 carbons, optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, carboxyalkyl having 1-8 carbons, or phenyl, where phenyl is optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, aminoalkyl having 1-8 carbons, or carboxyalkyl having 1-8 carbons;
or, one or more of R21, R22 and R23, taken in combination with SPACER or SPACER'"'"' or R20 forms a 5- or 6-membered aromatic, heteroaromatic, alicyclic or heteroalicyclic ring, the heteroatoms selected from O, N or S;
where Ψ
- is a biologically compatible counterion;or CAP and CAP'"'"' are independently ##STR141## where R21, R22 and Ψ
- are as defined previously;
such that at least one of R3, R4, R6, R7, R11, R12, R13, and R14 is a TAIL; and
, where more than one substituent is a TAIL, each TAIL is optionally the same or different;wherein the fluorescent complex has a set of characteristic spectral properties; b) combining the fluorescent complex with a sample that contains or is thought to contain an analyte that interacts with the nucleic acid polymer; c) detecting a change in the spectral properties of the fluorescent complex; and d) determining the presence or activity of the analyte in said sample based on comparison of the change in spectral properties of the complex with a fluorescent standard characteristic of analyte activity. - View Dependent Claims (76, 77, 78, 79)
-
-
80. A compound having the formula ##STR142## wherein R24 is a linear or branched alkyl having 1-6 carbons;
- and Ψ
- is a biologically compatible counterion.
- and Ψ
-
81. A compound of the formula ##STR143## wherein each R1 is independently H;
- or an alkyl group having from 1-6 carbons;
an alkoxy group having from 1-6 carbons;
or a trifluoromethyl;
or a halogen; and
t=1-4;R2 is an alkyl group having 1-6 carbons; X is O, S, Se or NR15, where R15 is an alkyl group having 1-6 carbons;
or X is CR16 R17 where R16 and R17, which may be the same or different, are independently alkyl groups having 1-6 carbons, or R16 and R17 taken in combination complete a five or six membered saturated ring;n=0, 1 or2; Ψ
- is a biologically compatible counterion;Q has the formula Q 1 or Q2 ##STR144## wherein Y is --CR3 ═
CR4 --;p and m=0 or 1, such that p+m=1; R5 is an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or R5 is a TAIL;R4 is an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;
where R8 is an alkyl group having 1-6 carbons, or 1-2 alicyclic or aromatic rings; and
R9 is H, an alkyl group having 1-6 carbons, or 1-2 alicyclic or aromatic rings;
or R8 and R9 taken in combination are --(CH2)4 -- or --(CH2)5 -- to give a 5 or 6 membered ring; and
where R19 is alkyl having 1-6 carbons, or perfluoroalkyl having 1-6 carbons, or aryl;R3, R6 and R7, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-10 carbons;
or --OR8, --SR8, --(NR8 R9);
or --OSO2 R19 ;
or a TAIL;or R6 and R7, taken in combination are --(CH2)v -- where v=3 or 4, or R6 and R7 form a fused aromatic ring according to formula Q2; R11, R12, R13, and R14, which may be the same or different, are independently H;
or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons;
or a halogen;
or a substituted or unsubstituted aryl or heteroaryl;
or a substituted or unsubstituted cycloalkyl having 3-8 carbons;
or a TAIL;
or --OR8, --SR8, or --(NR8 R9);TAIL is a heteroatom-containing moiety having the formula LINK-SPACER-CAP; wherein LINK is a single covalent bond, --O--, --S--, or --NR20 --;
where R20 is H, a linear or branched alkyl having 1-8 carbons, or R20 is --SPACER'"'"'--CAP'"'"';SPACER and SPACER'"'"', which may be the same or different are linear or branched, cyclic or heterocyclic, saturated or unsaturated covalent linkages, each having 1-16 nonhydrogen atoms selected from the group consisting of C, N, P, O and S, such that the linkage contains any combination of ether, thioether, amine, ester, amide bonds;
or single, double, triple or aromatic carbon-carbon bonds;
or phosphorus-oxygen, phosphorus-sulfur bonds, nitrogen-nitrogen or nitrogen-oxygen bonds;
or aromatic or heteroaromatic bonds;CAP and CAP'"'"', which may be the same or different, are --O--R21, --S--R21, --NR21 R22, or --N+ R21 R22 R23 Ψ
- ;wherein R21, R22, and R23 are independently H, or a linear or branched alkyl or cycloalkyl having 1-8 carbons, optionally further substituted by hydroxy, alkoxy having 1-8 carbons, carboxyalkyl having 1-8 carbons, or phenyl, where phenyl is optionally further substituted by halogen, hydroxy, alkoxy having 1-8 carbons, aminoalkyl having 1-8 carbons, or carboxyalkyl having 1-8 carbons;
or, one or more of R21, R22 and R23, taken in combination with SPACER or SPACER'"'"' or R20 forms a 5- or 6-membered aromatic, heteroaromatic, alicyclic or heteroalicyclic ring, the heteroatoms selected from O, N or S;
where Ψ
- is a biologically compatible counterion;or CAP and CAP'"'"' are independently ##STR145## where R21, R22 and Ψ
- are as defined previously;
such that when R5 is a TAIL, LINK is a single bond; and
, where more than one of R2, R3, R4, R5, R6, R7, R11, R12, R13, and R14 is a TAIL, each TAIL is optionally the same or different. - View Dependent Claims (82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97)
- or an alkyl group having from 1-6 carbons;
Specification