Phosphine derivatives of fluorescent compounds
A phosphine derivative of DyLight dyes modified with ethylene glycol or (poly)ethylene glycol groups. In one embodiment, the compounds are useful in chemoselective ligation reactions.
1. A compound of general formula III
where
each of R 1 , R 2 , R 5 , and R 6 is the same or different and is independently selected from the group consisting of an aliphatic, heteroaliphatic, sulfoalkyl group, or heteroaliphatic with terminal SO 3 ;
each of R 7 and R 8 is the same or different and is independently selected from either H, SO 3 , sulfoalkyl, heteroaliphatic, or heteroaliphatic with terminal SO 3 ;
each of R9 and R10 is the same or different and is independently selected from either aryl groups, substituted aryl groups, or cycloalkyl groups;
R11 is an electrophilic group selected from the group consisting of a carboxylic acid, an alkyl ester, an aryl ester, a substituted aryl ester, an aldehyde, and amide, an aryl amide, an alkyl halide, a thioester, a sulfonyl ester, and alkyl ketone, an aryl ketone, a substituted aryl ketone, a halosulfonyl, a nitrile, and a nitro;
L is a linking group selected from the group consisting of a divalent linear (—(CH 2 ) o —, o=1 to 15), crossed, cyclic, and combinations thereof alkylene group which can be substituted by at least one atom selected from the group consisting of oxygen, substituted nitrogen, and sulfur;
Kat is a number of Na + , K + , Ca 2+ , ammonia, or other cation(s) needed to compensate the negative charge brought by the cyanine;
m is an integer from 0 to 5 inclusive; o is an integer from 0 to 12 inclusive; and n is an integer from 1 to 3 inclusive.
2. A compound of general formula IV
where
each of R 1 , R 2 , R 5 , and R 6 is the same or different and is independently selected from the group consisting of an aliphatic, heteroaliphatic, sulfoalkyl group, or heteroaliphatic with terminal SO 3 ;
each of R 7 and R 8 is the same or different and is independently selected from either H, SO 3 , sulfoalkyl, heteroaliphatic, or heteroaliphatic with terminal SO 3 ;
each of R9 and R10 is the same or different and is independently selected from either aryl groups, substituted aryl groups, or cycloalkyl groups;
R11 is an electrophilic group selected from the group consisting of a carboxylic acid, an alkyl ester, an aryl ester, a substituted aryl ester, an aldehyde, and amide, an aryl amide, an alkyl halide, a thioester, a sulfonyl ester, and alkyl ketone, an aryl ketone, a substituted aryl ketone, a halosulfonyl, a nitrile, and a nitro;
L is a linking group selected from the group consisting of a divalent linear (—(CH 2 ) o —, o=1 to 15), crossed, cyclic, and combinations thereof alkylene group which can be substituted by at least one atom selected from the group consisting of oxygen, substituted nitrogen, and sulfur;
Kat is a number of Na + , K + , Ca 2+ , ammonia, or other cation(s) needed to compensate the negative charge brought by the cyanine;
m is an integer from 0 to 5 inclusive; p is an integer from 1 to 6 inclusive; and n is an integer from 1 to 3 inclusive.
3. A compound of formula
4. A compound of formula
5. A compound of formula
6. A method using the compound of claim 1 in a chemoselective ligation reaction with an azido compound.
7. A method using the compound of claim 2 in a chemoselective ligation reaction with an azido compound.
8. A method using the compound of claim 3 in a chemoselective ligation reaction with an azido compound.
9. A method using the compound of claim 4 in a chemoselective ligation reaction with an azido compound.
10. A method using the compound of claim 5 in a chemoselective ligation reaction with an azido compound.
11. A method of labeling at least one biomolecule, the method comprising providing a composition comprising at least one excipient and the compound of claim 1 in an effective concentration to the biomolecule under conditions sufficient for specifically binding the compound to the biomolecule, in the absence of non-specific binding, resulting in labeling of the biomolecule.
12. A method of labeling at least one biomolecule, the method comprising providing a composition comprising at least one excipient and the compound of claim 2 in an effective concentration to the biomolecule under conditions sufficient for specifically binding the compound to the biomolecule, in the absence of non-specific binding, resulting in labeling of the biomolecule.
13. The method of claim 11 further comprising reacting a linking moiety with the at least one biomolecule to result in an at least one biomolecule-linking moiety conjugate, and reacting the compound with the at least one biomolecule-linking moiety conjugate to result in an at least one biomolecule-linking moiety-compound conjugate.
14. The method of claim 12 further comprising reacting a linking moiety with the at least one biomolecule to result in an at least one biomolecule-linking moiety conjugate, and reacting the compound with the at least one biomolecule-linking moiety conjugate to result in an at least one biomolecule-linking moiety-compound conjugate.
15. The method of claim 13 where the linking moiety is a heterobifunctional linking moiety having a first terminal group reactive with an amine group and a second terminal azide.
16. The method of claim 14 where the linking moiety comprises a polyethylene glycol (PEG) group between the first and the second functional termini.
17. A method of detecting at least one biomolecule, the method comprising providing a composition comprising at least one excipient and the compound of claim 1 in an effective concentration to at least one biomolecule under conditions sufficient for binding the compound to the biomolecule, and detecting the biomolecule-bound compound.
18. A method of detecting at least one biomolecule, the method comprising providing a composition comprising at least one excipient and the compound of claim 2 in an effective concentration to at least one biomolecule under conditions sufficient for binding the compound to the biomolecule, and detecting the biomolecule-bound compound.
19. A kit comprising a compound of claim 1 and instructions for performing a chemoselective ligation reaction using the compound with an azido compound.
20. A kit comprising a compound of claim 2 and instructions for performing a chemoselective ligation reaction using the compound with an azido compound.