Ultra bright dimeric or polymeric dyes with spacing linker groups
Compounds useful as fluorescent or colored dyes are disclosed. The compounds have the following structure (I): or a stereoisomer, tautomer or salt thereof, wherein R 1 , R 2 , R 3 , R 4 , R 5 , L 1 , L 2 , L 3 , L 4 , M, m and n are as defined herein. Methods associated with preparation and use of such compounds are also provided.
1. A compound having the following structure (IB):
or a stereoisomer or salt thereof wherein:
M is, at each occurrence, independently a fluorescent dye;
R 1 is, at each occurrence, H;
R 2 is a linker comprising a covalent bond to a targeting moiety;
R 3 is OH, SH, —OP(═R a )(R b )R c or L′;
R 4 is, at each occurrence, independently OH, SH, O − , S − , OR d or SR d ;
R 5 is, at each occurrence, independently oxo or thioxo;
R a is O or S;
R b is OH, SH, O − , S − , OR d or SR d ;
R c is OH, SH, O − , S − , OR d , OL′ or SR d ;
R d is a cation;
L′ is a linker comprising a covalent bond to a solid support, a linker comprising a covalent bond to a solid support residue or a linker comprising a covalent bond to a nucleoside;
L 1 is at each occurrence, independently an alkylene or heteroalkylene linker;
m is, at each occurrence, independently an integer of one or greater;
n is an integer of one or greater;
x 1 , x 2 , x 3 and x 4 are, at each occurrence, independently an integer from 0 to 6; and
z is an integer from 2 to 100.
2. The compound of claim 1 , wherein the targeting moiety selectively binds to a biomolecule.
3. The compound of claim 1 , wherein the biomolecule is an enzyme, receptor, receptor ligand, antibody, bacteria, virus, blood cell or tissue cell.
4. The compound of claim 1 , wherein x 1 , x 2 , x 3 and x 4 are each 1 at each occurrence.
5. The compound of claim 1 , wherein R 4 is, at each occurrence, independently OH, O − or OR d , and R 5 is, at each occurrence, oxo.
6. The compound of claim 1 , wherein R 3 is OH.
7. The compound of claim 1 , wherein R 3 is —OP(═R a )(R b )R c .
8. The compound of claim 7 , wherein R c is OL′.
9. The compound of claim 8 , wherein L′ has the following structure:
wherein:
m″ and n″ are independently an integer from 1 to 10;
R e is H, an electron pair or a counter ion; and
L″ is R e or a direct bond or linkage to a solid support, a solid support residue or a nucleoside.
10. The compound of claim 1 , wherein R 3 has the following structure:
11. The compound of claim 1 , wherein R 3 has the following structure:
12. The compound of claim 1 , wherein L 1 , at each occurrence, independently has one of the following structures:
13. The compound of claim 1 , wherein m is, at each occurrence, independently an integer from 1 to 10.
14. The compound of claim 1 , wherein m is, at each occurrence, independently 3, 4, 5 or 6, and z is an integer from 3 to 6.
15. The compound of claim 1 , wherein n is an integer from 1 to 10.
16. The compound of claim 1 , wherein z is an integer from 3 to 10.
17. The compound of claim 1 , wherein z is an integer from 20 to 25.
18. The compound of claim 1 , wherein M is, at each occurrence, independently pyrene, perylene, perylene monoimide or 6-FAM or derivative thereof.
19. The compound of claim 1 , wherein M, at each occurrence, independently has one of the following structures:
20. The compound of claim 1 , having the following structure:
wherein:
z is 3, 4, 5 or 6;
m is 2, 3 4 or 5;
n is an integer from 1 to 10; and
F″ has the following structure:
21. A composition comprising the compound of claim 1 and one or more biomolecules.
22. The composition of claim 21 , wherein the biomolecule is an enzyme, receptor, receptor ligand, antibody, bacteria, virus, blood cell or tissue cell.
23. A method of staining a sample, comprising adding to the sample the compound of claim 1 in an amount sufficient to produce an optical response when said sample is illuminated at an appropriate wavelength.
24. A method for visually detecting biomolecule, the method comprising:
(a) admixing the compound of claim 1 and the biomolecule, thereby associating the compound and the analyte; and
(b) detecting the compound by its visible properties.