Quinone methide analog signal amplification

Patent metadata

Field	Value
Publication number	US-11780863-B2
Application number	US-201816183583-A
Country	US
Kind code	B2
Filing date	Nov 7, 2018
Priority date	Feb 24, 2014
Publication date	Oct 10, 2023
Grant date	Oct 10, 2023

Abstract

Disclosed herein are novel quinone methide analog precursors and embodiments of a method and a kit of using the same for detecting one or more targets in a biological sample. The method of detection comprises contacting the sample with a detection probe, then contacting the sample with a labeling conjugate that comprises an enzyme. The enzyme interacts with a quinone methide analog precursor comprising a detectable label, forming a reactive quinone methide analog, which binds to the biological sample proximally to or directly on the target. The detectable label is then detected. In some embodiments, multiple targets can be detected by multiple quinone methide analog precursors interacting with different enzymes without the need for an enzyme deactivation step.

First claim

The invention claimed is:

1. A compound, having formula (III):

wherein
each Q is Carbon;
each R 7 is independently selected from hydrogen, —Z—R 1 , a moiety comprising a LG group, —C(LG)(R 5 )(R 3 R 4 ), —R 3 R 4 , a moiety comprising a LG group and a detectable label, a halo group, an aliphatic group, an alkoxy group, or where two adjacent R 7 groups together form an aliphatic ring or aryl ring where the aliphatic ring or aryl ring formed from the two adjacent R 7 groups is substituted with at least an LG group and a moiety comprising a detectable label;
—Z—R 1 is —OP(O)(OH) 2 , a phosphate ester, or —O-sugar;
LG is a halide, alkoxy, carboxylate, inorganic ester, thiolate, amine, carboxylate, azide, sulfate ester, aryloxy, phenoxide or —N(R b ) 3 + where each R b is independently hydrogen or a lower alkyl or two R b moieties together form a heteroaliphatic ring;
R 3 is independently —(CH 2 ) n NH—, —O(CH 2 ) n NH—, —N(H)C(O)(CH 2 ) n NH—, —C(O)N(H)(CH 2 ) n NH—, —(CH 2 ) n O—, —O(CH 2 ) n O—, —O(CH 2 CH 2 O) n —, —N(H)C(O)(CH 2 ) n O—, —C(O)N(H)(CH 2 ) n O—, C(O)N(H)(CH 2 CH 2 O) n —, —(CH 2 ) n S—, —O(CH 2 ) n S—, —N(H)C(O)(CH 2 ) n S—, —C(O)N(H)(CH 2 ) n S—, —(CH 2 ) n NH—, —C(O)N(H)(CH 2 CH 2 O) n CH 2 CH 2 NH—, —C(O)(CH 2 CH 2 O) n CH 2 CH 2 NH—, —C(O)N(H)(CH 2 ) n NHC(O)CH(CH 3 )(CH 2 ) n NH—, or —N(H)(CH 2 ) n NH—, where each n independently is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12;
m is 1,
at least one of the R 7 groups comprises —Z—R 1 ;
R 4 is a detectable label selected from a hapten, an annulated chromophore, a chromogen capable of being visualized using bright-field illumination, a diazo dye, a diazo-containing chromogen, biotin, fluorescein, fluorescein derivatives, a coumarin, a coumarin derivative, a tartrazine, or a triarylmethane;
R 5 is H;
wherein the compound of Formula (III) comprises at least one detectable label; and
wherein the compound of Formula (III) comprises at least two adjacent R 7 groups, wherein the at least two adjacent R 7 groups comprise an indoline group.

2. The compound of claim 1 , wherein Z—R 1 is —O—P(O)(OH) 2 .

3. The compound of claim 1 , wherein the compound has the structure:

4. The compound of claim 1 wherein the compound has the structure:

5. The compound of claim 1 , wherein the detectable label is selected from the group consisting of benzodiazepine, benzofurazan, 4-(dimethylamino)azobenzene-4′-sulfonamide, 7-(diethylamino)coumarin-3-carboxylic acid), digoxigenin, dinitrophenyl, 3-hydroxy-2-quinoxalinecarbamide, nitrocinnamic acid, nitropyrazole, Podophyllotoxin, rhodamine, rotenone, and thiazolesulfonamide.

6. The compound of claim 1 wherein Z—R 1 is —O-sugar.

7. The compound of claim 1 , wherein the detectable label comprises at least one PEG group.

8. A compound selected from the group consisting of

R 4 is a detectable label selected from a hapten, an annulated chromophore, a chromogen capable of being visualized using bright-field illumination, a diazo dye, a diazo-containing chromogen, biotin, fluorescein, fluorescein derivatives, a coumarin, a coumarin derivative, a tartrazine, or a triarylmethane.

9. The compound of claim 8 , wherein the compound has the structure:

10. A compound selected from the group consisting of:

wherein —Z—R 1 is —OP(O)(OH) 2 , a phosphate ester, or —O-sugar; and wherein R 4 is selected from a hapten, an annulated chromophore, a chromogen capable of being visualized using bright-field illumination, a diazo dye, a diazo-containing chromogen, biotin, fluorescein, fluorescein derivatives, a coumarin, a coumarin derivative, a tartrazine, or a triarylmethane.

Assignees

• Ventana Med Syst Inc

Inventors

• Bieniarz Christopher
• Ashworth-Sharpe Julia
• Kelly Brian D
• Polaske Nathan

Classifications

• C07F9/6561 Primary

  containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings · CPC title

• C07D209/14

  Radicals substituted by nitrogen atoms, not forming part of a nitro radical · CPC title

• C07D403/06

  linked by a carbon chain containing only aliphatic carbon atoms · CPC title

• C07F9/06

  without P—C bonds · CPC title

• C07F9/12

  with hydroxyaryl compounds · CPC title