HETEROCYCLE-SUBSTITUTED XANTHENE DYES
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Abstract
The present invention relates to fluorescent dyes in general. The present invention provides a wide range of fluorescent dyes and kits containing the same, which are applicable for labeling a variety of biomolecules, cells and microorganisms. The present invention also provides various methods of using the fluorescent dyes for research and development, forensic identification, environmental studies, diagnosis, prognosis, and/or treatment of disease conditions.
95 Citations
81 Claims
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1. A compound of Formula 1a or 1b:
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2. The compound of claim 1, wherein X is O.
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3. The compound of claim 1, wherein A is —
- NR1R1a and B is ═
N R4R4a.
- NR1R1a and B is ═
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4. The compound of claim 1, wherein A is —
- NR1R1a and B is ═
N R4R4a, and wherein at least one pair of R1 and R9, or R4 and R5 taken together with the atoms to which they are attached form one or more fused saturated or unsaturated rings that are optionally substituted by at least one halogen, CN, alkyl, alkenyl, alkoxy, alkylthio, alkylamino, dialkylamino, alkylaminocarbonyl, dialkylaminocarbonyl, alkylaminosulfonyl, dialkylaminosulfonyl, aminosulfonyl, aryl, heteroaryl, -L-PO32−
, L-SO3−
, or -L-Rx.
- NR1R1a and B is ═
-
5. The compound of claim 4, wherein said at least one ring is substituted by -L-PO32−
- or -L-SO3−
.
- or -L-SO3−
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6. The compound of claim 4, wherein said at least one ring is substituted by at least one alkyl.
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7. The compound of claim 1, wherein at least one of R2 and R3 is -L-SO3−
- .
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8. The compound of claim 1, wherein R7 is
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9. The compound of claim 8, wherein R10 and R11 are H.
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10. (canceled)
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11-21. -21. (canceled)
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22. The compound of claim 1, wherein at least one of R12 and R13 is -L-Rx.
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23. The compound of claim 1, wherein at least one of R12 and R13 is -L-SO3−
- .
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24-32. -32. (canceled)
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33. A compound of Formula 1a or 1b:
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34-41. -41. (canceled)
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42. A kit comprising:
i) a compound of claim 1; and
ii) instructions instructing the use of the compound.
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43. The kit of claim 42, wherein the kit additionally comprises a buffer and optionally materials or devices for purifying conjugation products.
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44. A biomolecule comprising a label having a structure of a compound of claim 1 comprising at least one reactive moiety, wherein the at least one reactive moiety of the compound has undergone a reaction which attaches the label to the biomolecule.
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45. The biomolecule comprising a label of claim 44 wherein the biomolecule comprises a polynucleotide, a polypeptide, a polypeptide further comprising an antigen binding site, a whole immunoglobulin, or a Fab fragment.
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46-49. -49. (canceled)
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50. An immunoglobin comprising a label having a structure of claim 1 comprising at least one reactive moiety, wherein at least one reactive moiety of the compound has undergone a reaction which attaches the label to the immunoglobin, wherein the immunoglobin is an antibody that binds specifically to an antigen on a cancer cell.
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51. The immunoglobin of claim 50 wherein the antibody binds to erb2.
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52. A method of preparing a labeled biomolecule comprising reacting a compound of claim 1 comprising at least one reactive moiety with a substrate biomolecule under conditions sufficient to effect crosslinking between the compound and the substrate biomolecule.
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53. The method of claim 52, wherein the substrate biomolecule is a protein, polypeptide, a polynucleotide, a carbohydrate, a lipid, a metal chelator or a combination thereof.
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54-66. -66. (canceled)
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67. A method of labeling a polypeptide comprising:
- forming a complex that comprises the polypeptide and a binding agent, wherein the binding agent comprises a fluorescent label having a structure of claim 1 comprising at least one reactive moiety, wherein the at least one reactive moiety has undergone a reaction which attaches the label to the binding agent.
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68-74. -74. (canceled)
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75. A compound of formula 1a or formula 1b:
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76. A compound of formula 1a or formula 1b:
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77. The compound of claim 76, wherein R7 is a 5-membered heterocycle.
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78. A biomolecule according to claim 45, wherein said biomolecule comprises a fluorescent polypeptide.
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79. The biomolecule according to claim 78, wherein said fluorescent polypeptide is selected from the group consisting of R-PE, APC, and PerCP.
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80. The compound of claim 1, wherein R12 is independently alkyl, alkenyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylaminosulfonyl, dialkylaminosulfonyl, aminosulfonyl, aryl, heteroaryl, -L-PO32−
- , L-SO3−
, or -L-Rx.
- , L-SO3−
-
81. The compound of claim 1, wherein said compound is selected from the group consisting of
Specification