Xanthene dyes comprising a sulfonamide group

• US 9,006,437 B2
• Filed: 04/09/2013
• Issued: 04/14/2015
• Est. Priority Date: 02/03/2009
• Status: Active Grant

• Current Assignee: Biotium, Inc.
• Original Assignee: Biotium, Inc.
• Inventors: Mao, Fei, Leung, Wai-Yee, Cheung, Ching-Ying
• Primary Examiner(s): VAJDA, KRISTIN ANN
• Application Number:
  US13/859,627
• Publication Number:
  US 20140011208A1
• Time in Patent Office: 735 Days
• Field of Search: 435/7.1, 544/357, 544/405, 536/23.1, 530/391.3, 530/408, 546/256, 546/48, 549/226, 548/525
• US Class Current: 544/357
• CPC Class Codes: C07D 311/88 Nitrogen atoms
• CPC Class Codes: C07D 405/12 linked by a chain containing hetero atoms as chain links
• CPC Class Codes: C07D 405/14 containing three or more hetero rings
• CPC Class Codes: C07D 491/147 the condensed system containing one ring with oxygen as ring hetero atom and two rings with nitrogen as ring hetero atom
• CPC Class Codes: C07D 491/22 in which the condensed system contains four or more hetero rings
• CPC Class Codes: C07K 1/13 Labelling of peptides
• CPC Class Codes: C09B 11/24 Phthaleins containing amino groups ; Phthalanes; Fluoranes; Phthalides; Rhodamine dyes; Phthaleins having heterocyclic aryl rings; Lactone or lactame forms of triarylmethane dyes
• CPC Class Codes: G01N 33/57484 involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
• CPC Class Codes: G01N 33/582 with fluorescent label

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.

Claims

1. A compound of Formula Ia or Ib:

2. The compound of claim 1, wherein R₁₀ is H, sulfopropyl or sulfobutyl.

3. The compound of claim 1, wherein R_x is an isothiocyanate, an isocyanate, a monochlorotriazine, a dichlorotriazine, a halogen-substituted pyridine, a halogen-substituted diazine, a phosphoramidite, a maleimide, an aziridine, a sulfonyl halide, an acid halide, a hydroxysuccinimidyl ester, a hydroxysulfosuccinimidyl ester, a pyrrole-2,5-dione, a tetrafluorophenol ester, an imido ester, a hydrazine, an azidonitrophenyl, an azide, an alkyne, a 3-(2-pyridyl dithio)-propionamide, a glyoxal or an aldehyde.

4. The compound of claim 1, wherein R_x forms a covalent bond with an amino, a sulfhydryl or a hydroxy nucleophile.

5. The compound of claim 1, wherein R₂, R₃, R₅ and R₉ is independently selected from the group consisting of H, halogen, —

PO₃²⁻

, —

SO₃⁻

or unreactive sulfonamide, or

6. The compound of claim 1, wherein R₇ is H, substituted or unsubstituted alkyl, or substituted or unsubstituted aryl.

7. The compound of claim 1, wherein R₇ is a phenyl substituted with —

CO₂⁻

, carboxamide, —

SO₃⁻

, chloro, fluoro or R_x.

8. The compound of claim 1, wherein R_x is an activated carboxylic acid ester group.

9. The compound of claim 8, wherein the activated carboxylic acid ester group is an N-hydroxysuccinimide ester group.

10. The compound of claim 1 having the structure of Formula IIIa:

11. The compound of claim 10 wherein:

R₃ is H or —

SO₃⁻

; and

L is Q_n, wherein at least one, two, three or four Q are —

(CH₂CH₂O)—

.

12. A kit comprising:

i) a compound of claim 1;

ii) a buffer;

iii) materials or devices for purifying conjugation products; and

iv) instructions instructing the use of the compound.