Quinone-masked probes as labeling reagents for cell uptake measurements

Patent metadata

Field	Value
Publication number	US-9790537-B2
Application number	US-201514609372-A
Country	US
Kind code	B2
Filing date	Jan 29, 2015
Priority date	Jan 29, 2014
Publication date	Oct 17, 2017
Grant date	Oct 17, 2017

Abstract

Provided are labeling reagents and methods of using the reagents for cell uptake measurements. The labeling reagents can be quinone-masked probes including fluorophores and/or luminophores.

First claim

What is claimed is:

1. A compound of formula (I), or a salt thereof,

wherein
A is a reporter moiety;
R 14 is H, alkyl, hydroxyalkyl, alkoxy, carboxyalkyl, or amidoalkyl;
R 9 and R 10 are independently selected from alkyl;
R 11 , R 12 and R 13 are independently selected from alkyl;
X is O;
L is —(CH 2 ) m C(R 17 ) 2 (CH 2 ) n —Y—C(O)—;
R 17 is independently H, alkyl or both R 17 together can form an alkyl ring having from 3-7 carbons;
m is an integer from 0-2;
n is an integer from 0-2;
Y is O or NR 15 ;
R 5 is -alkyl-amide, wherein the -alkyl-amide is -alkyl-CON(R 32 )(R 33 ) or -alkyl-(CO)—NR 34 —(CR a R b ) p —NR 35 (CO)-T, wherein
R 32 and R 33 are each independently selected from hydrogen, alkyl, carboxy, aryl, arylalkyl, cycloalkylalkyl, heteroaryl, heterocycle, alkene, polyol, alkenylpolylol, a peptide, a drug, a derivative of a drug, a biologically active moiety, and a dye;
R 34 and R 35 are each independently selected from hydrogen, and alkyl;
R a and R b are each independently selected from hydrogen, alkyl, and carboxy;
p is 0 to 6; and
T is selected from aryl, arylalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, heterocycle, hydroxyalkyl, protein, polypeptide, polypeptide-based toxin, amino acid, nucleotide, polynucleotide, lipid, sugar, carbohydrate, enzyme substrate, drug, derivative of a drug, polymer-linked nanoparticle, antibody, detergent, and a dye, or a combination thereof;

or
R 15 is polyalkoxyalkyl, wherein the polyalkoxyalkyl is —(C 2 -C 6 -alkoxy) x -alkyl-CON(R 32 )(R 33 ) or —(C 2 -C 6 -alkoxy) x -alkyl-(CO)—NR 34 -(CR a R b ) p —NR 35 (CO)-T;
x is an integer selected from 1 to 20;
R 32 and R 33 are each independently selected from hydrogen, alkyl, carboxy, aryl, arylalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, heterocycle, hydroxyalkyl, protein, polypeptide, polypeptide-based toxin, amino acid, nucleotide, polynucleotide, lipid, sugar, carbohydrate, enzyme substrate, drug, derivative of a drug, polymer-linked nanoparticle, antibody, detergent, and a dye, or a combination thereof;
R 34 and R 35 are each independently selected from hydrogen, and alkyl;
R a and R b are each independently selected from hydrogen, alkyl, and carboxy;
p is 0 to 6; and
T is selected from aryl, arylalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, heterocycle, hydroxyalkyl, protein, polypeptide, polypeptide-based toxin, amino acid, nucleotide, polynucleotide, lipid, sugar, carbohydrate, enzyme substrate, drug, derivative of a drug, polymer-linked nanoparticle, antibody, detergent, and a dye, or a combination thereof;

or
R 15 is

wherein
R 40 is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, and cycloalkyl, wherein said aryl, heteroaryl, heterocyclyl, and cycloalkyl are unsubstituted or substituted with one or more suitable substituents; and
x is an integer selected from 1 to 20.

2. The compound of claim 1 , wherein R 15 is polyalkoxyalkyl, wherein the polyalkoxyalkyl is —(C 2 -C 6 -alkoxy) x -alkyl-CON(R 32 )(R 33 ) or —(C 2 -C 6 -alkoxy) x -alkyl-(CO)—NR 34 —(CR a R b ) p —NR 35 (CO)-T;
x is an integer selected from 1 to 20;
R 32 and R 33 are each independently selected from hydrogen, alkyl, carboxy, aryl, arylalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, heterocycle, hydroxyalkyl, protein, polypeptide, polypeptide-based toxin, amino acid, nucleotide, polynucleotide, lipid, sugar, carbohydrate, enzyme substrate, drug, derivative of a drug, polymer-linked nanoparticle, antibody, detergent, and a dye, or a combination thereof;
R 34 and R 35 are each independently selected from hydrogen, and alkyl;
R a and R b are each independently selected from hydrogen, alkyl, and carboxy;
p is 0 to 6; and
T is selected from aryl, arylalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, heterocycle, hydroxyalkyl, protein, polypeptide, polypeptide-based toxin, amino acid, nucleotide, polynucleotide, lipid, sugar, carbohydrate, enzyme substrate, drug, derivative of a drug, polymer-linked nanoparticle, antibody, detergent, and a dye, or a combination thereof.

3. The compound of claim 1 , wherein R 15 is

wherein
T is selected from aryl, arylalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, heterocycle, hydroxyalkyl, protein, polypeptide, polypeptide-based toxin, amino acid, nucleotide, polynucleotide, lipid, sugar, carbohydrate, enzyme substrate, drug, derivative of a drug, polymer-linked nanoparticle, antibody, detergent, and a dye, or a combination thereof; and
x is an integer selected from 1 to 20.

4. The compound of claim 1 , wherein R 15 is -alkyl-amide, wherein the -alkyl-amide is -alkyl-CON(R 32 )(R 33 ) or -alkyl-(CO)—NR 34 —(CR a R b ) p —NR 35 (CO)-T, wherein
R 32 and R 33 are each independently selected from hydrogen, alkyl, carboxy, aryl, arylalkyl, cycloalkylalkyl, heteroaryl, heterocycle, alkene, polyol, alkenylpolylol, a peptide, a drug, a derivative of a drug, a biologically active moiety, and a dye;
R 34 and R 35 are each independently selected from hydrogen, and alkyl;
R a and R b are each independently selected from hydrogen, alkyl, and carboxy;
p is 0 to 6; and
T is selected from aryl, arylalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, heterocycle, hydroxyalkyl, protein, polypeptide, polypeptide-based toxin, amino acid, nucleotide, polynucleotide, lipid, sugar, carbohydrate, enzyme substrate, drug, derivative of a drug, polymer-linked nanoparticle, antibody, detergent, and a dye, or a combination thereof.

5. The compound of claim 1 , wherein R 5 is

wherein
T is selected from aryl, arylalkyl, cycloalkylalkyl, heteroaryl, heterocycle, hydroxyalkyl, protein, polypeptide, polypeptide-based toxin, amino acid, nucleotide, polynucleotide, lipid, sugar, carbohydrate, enzyme substrate, drug, derivative of a drug, polymer-linked nanoparticle, antibody, detergent, and a dye, or a combination thereof; and
x is 0 to 20.

6. The compound of claim 1 , wherein R 15 is

wherein
R 40 is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, and cycloalkyl, wherein said aryl, heteroaryl, heterocyclyl, and cycloalkyl are unsubstituted or substituted with one or more suitable substituents; and
x is an integer selected from 1 to 20.

7. The compound of claim 6 , wherein R 40 is a 5- or 6-membered heterocyclyl, having 1, 2, or 3 heteroatoms independently selected from the group consisting of N, O, and S.

8. The compound of claim 6 , having formula (I-vi), or a salt thereof,

9. The compound of claim 8 , wherein
R 9 , R 10 , R 11 , R 12 , R 13 , and R 14 are methyl;
x is 3;
R 40 is hydrogen; and
A is

10. The compound of claim 1 ,

Assignees

• Promega Corp

Inventors

• Zhou Wenhui
• Meisenheimer Poncho
• Duellman Sarah
• Ma Dongping
• Cali James J
• Mustafa Dana

Classifications

• C12Q1/66 Primary

  involving luciferase · CPC title

• G01N33/582

  with fluorescent label · CPC title

• C07D417/14

  containing three or more hetero rings · CPC title

• C07K16/40

  against enzymes · CPC title

• C07J43/003

  not condensed · CPC title