Carbopyronone compounds useful as diagnostic adjuvants

Patent metadata

Field	Value
Publication number	US-10018617-B2
Application number	US-201515304800-A
Country	US
Kind code	B2
Filing date	May 15, 2015
Priority date	May 16, 2014
Publication date	Jul 10, 2018
Grant date	Jul 10, 2018

Abstract

Described herein are fluorescent compounds and methods and comprising these compounds. The compounds disclosed herein are carbopyronine reagents that fluoresce in the red portion of the UV/VIS spectrum and provide bright fluorescence intensity, uniform cell staining, and good retention within live cells as well as low toxicity toward cells.

First claim

We claim:

1. A method for tracking cell proliferation and differentiation, the method comprising the steps of:
a) incubating a mixture of cells and a compound of structural formula (II):

or a pharmaceutically acceptable salt thereof,
wherein
R 12 and R 13 are each independently H, C 1 -C 6 substituted alkyl, -L-R x , or -L-S c ;
R 2 is carboxyl, -L-R x , or -L-S c ;
R 3 , R 4 , R 5 and R 6 are each independently H or halogen;
R 7 and R 9 are each C 1 -C 6 alkyl, C 1 -C 6 substituted alkyl, sulfoalky, -L-R x , or -L-S c ;
R 11 , R 14 , R 17 , R 18 , R 19 , R 20 , R 21 and R 22 are each H;
L is a linker;
R x is an acrylamide, a carboxylic acid, an activated ester of a carboxylic acid, an acyl azide, an acyl halide, hydroxy, an aldehyde, an alkyl halide, a sulfonate, an amine, an anhydride, an aniline, an aryl halide, an azide, an aziridine, a boronate, a carbodiimide, a diazoalkane, an epoxide, a glycol, a haloacetamide, a halotriazine, a hydrazine, a hydroxylamine, an imido ester, an isothiocyanate, a ketone, a maleimide, a sulfonyl halide, a thiol group, a succinimidyl ester (SE), a sulfodichlorophenyl (SDP) ester, a sulfotetrafluorophenyl (STP) ester, a tetrafluorophenyl (TFP) ester, a pentafluorophenyl (PFP) ester, a nitrilotriacetic acid (NTA), an aminodextran, and a cyclooctyne-amine; and
S c is a solid support, a tyramide, an amino acid, a peptide, a protein, a monosaccharide, a polysaccharide, an ion-complexing moiety, a nucleotide, an oligonucleotide, a nucleic acid, a hapten, a drug, a toxin, a lipid, a phospholipid, a lipoprotein, a lipopolysaccharide, a liposome, a lipophilic polymer, a PEG group, a non-biological organic polymer, a polymeric microparticle, an animal cell, a plant cell, a bacterium, a yeast, or a virus;
b) providing a stimulus to the mixture to elicit a fluorescent signal; and
c) analyzing the stimulated mixture.

2. The method according to claim 1 , further comprising a second compound excitable at a different wavelength than the compound of structural formula.

3. The method according to claim 2 , wherein the second compound is CFDA-SE or GFP.

4. The method according to claim 1 , wherein for the compound of structural formula R 12 and R 13 are each methyl.

5. The method according to claim 1 , wherein the compound is selected from the group consisting of:

Assignees

• Life Technologies Corp

Inventors

• Gee Kyle
• Kang Hee Chol
• Zhou Wenjun
• Dubbels Bradley
• Olszowy Michael
• O'Grady Michael

Classifications

• C07D471/04 Primary

  Ortho-condensed systems · CPC title

• C09B11/28

  Pyronines {; Xanthon, thioxanthon, selenoxanthan, telluroxanthon dyes} · CPC title

• G01N33/5014

  for testing toxicity · CPC title

• C07C309/61

  the carbon skeleton being further substituted by nitrogen atoms, not being part of nitro or nitroso groups · CPC title

• C07K16/2812

  against CD4 · CPC title