Fluorescent detection of amines and hydrazines and assaying methods thereof

Patent metadata

Field	Value
Publication number	US-12060508-B2
Application number	US-202218062492-A
Country	US
Kind code	B2
Filing date	Dec 6, 2022
Priority date	Oct 3, 2016
Publication date	Aug 13, 2024
Grant date	Aug 13, 2024

Abstract

Provided herein are processes for preparing fluorescent 1-cyano-2-substituted isoindole compounds or N-substituted phthalazinium compounds, comprising reacting an aromatic dialdehyde or aromatic aldehyde-ketone compound with a material that contains primary amino or hydrazine groups, and assaying methods involving the processes thereof.

First claim

What is claimed is:

1. A process for preparing a compound of Formula (II):

comprising reacting a compound of Formula (III):

with a compound of R—NHNH 2 ,
wherein:
X is CR a or N,
R 1 and R 2 are R a ; or R 1 and R 2 together with the carbon atoms they are attached to form a monocyclic or multicyclic fused aryl or heteroaryl ring, which is optionally substituted with one or more R a1 ;
R 3 is hydrogen, alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of which is optionally substituted with one or more R b ;
R is a moiety, which is attached to a solid support directly or via a linker;
each instance of R a is independently hydrogen, halogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, hydroxyl, alkoxy, cycloalkoxy, heterocyclyloxy, aryloxy, heteroaryloxy, cyano, nitro, amino, amido, carboxyl, sulfonyl, sulfonamido, acyl, acyloxy, alkoxycarbonyl, or phosphate; each of which is optionally substituted with one or more R b ;
each instance of R a1 is independently halogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, hydroxyl, alkoxy, cycloalkoxy, heterocyclyloxy, aryloxy, heteroaryloxy, cyano, nitro, amino, amido, carboxyl, sulfonyl, sulfonamido, acyl, acyloxy, alkoxycarbonyl, or phosphate; each of which is optionally substituted with one or more R b ;
each instance of R b is independently halogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, hydroxyl, alkoxy, cycloalkoxy, heterocyclyloxy, aryloxy, heteroaryloxy, cyano, nitro, amino, amido, carboxyl, sulfonyl, sulfonamido, acyl, acyloxy, alkoxycarbonyl, or phosphate; each of which is optionally substituted with one or more halogen, alkyl, or alkoxy; and
wherein the reaction between the compound of Formula (III) and R—NHNH 2 occurs under a mildly basic condition.

2. The process of claim 1 , wherein R is optionally substituted alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is attached to the solid support directly or via a linker.

3. The process of claim 1 , wherein R is optionally substituted alkyl.

4. The process of claim 1 , wherein R is optionally substituted phenyl.

5. The process of claim 1 , wherein R is optionally substituted 5- or 6-membered monocyclic heteroaryl, or optionally substituted 9- or 10-membered bicyclic fused heteroaryl.

6. The process of claim 5 , wherein R is optionally substituted pyrimidyl, optionally substituted triazinyl, or optionally substituted purinyl.

7. The process of claim 1 , wherein R is optionally substituted aminocarbonyl (NH—C═O) or optionally substituted alkylaminocarbonyl (alkyl-NH—C═O), each of which is attached to the solid support directly or via a linker.

8. The process of claim 7 , wherein R is C 2-6 alkyl-NHC(O)— or is propyl-NHC(O)—.

9. The process of claim 1 , wherein the compound of R—NHNH 2 is attached to the solid support, and together with the solid support comprises a compound of one of the following structures, or tautomers thereof:

wherein [SS] indicates the solid support and the strand moiety is a nucleic acid molecule.

10. The process of claim 9 , wherein the nucleic acid molecule is a DNA.

11. The process of claim 1 , wherein R 1 and R 2 together with the carbon atoms they are attached to form an optionally substituted phenyl ring or optionally substituted naphthalenyl ring.

12. The process of claim 1 , wherein R 1 and R 2 are each R a , and wherein each instance of R a is independently hydrogen, halo, cyano, alkyl, hydroxyl, or alkoxy.

13. The process of claim 1 , wherein R 3 is hydrogen, alkyl or phenyl.

14. The process of claim 1 , wherein the compound of Formula (III) is:

15. The process of claim 1 , wherein the compound of Formula (III) is:

wherein n is 0, 1, 2, 3, or 4; and m is 0, 1, 2, 3, or 4.

16. The process of claim 1 , wherein the compound of Formula (III) is:

17. The process of claim 1 , wherein the solid support is one or more of glass, modified or functionalized glass, plastics, polysaccharides, nylon, nitrocellulose, resins, silica, silicon, modified silicon, carbon, metals, inorganic glasses, and optical fiber bundles.

18. The process of claim 1 , wherein the solid support is a bead.

19. The process of claim 18 , wherein the bead is one or more of spherical silica beads, inorganic nanoparticles, magnetic nanoparticles, cadmium based dots, and cadmium free dots.

20. The process of claim 1 , further comprising:
detecting and measuring the fluorescence of the compound of Formula (II) to assay the compound of R—NHNH 2 .

21. The process of claim 1 , wherein the reaction between the compound of Formula (III) and R—NHNH 2 occurs at a pH from about 9 to about 10.

22. The process of claim 1 , wherein the reaction between the compound of Formula (III) and R—NHNH 2 occurs in a mixture of a water miscible solvent and a buffer solution.

Assignees

• Illumina Inc

Inventors

• Asadi Ali
• Li jian-sen

Classifications

• G01N2021/3155

  Measuring in two spectral ranges, e.g. UV and visible · CPC title

• G01N21/6428

  Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53) · CPC title

• G01N21/314

  with comparison of measurements at specific and non-specific wavelengths (dual wavelength spectrometry G01J3/427) · CPC title

• C09K2211/1022

  bridged by heteroatoms, e.g. N, P, Si or B · CPC title

• C07D471/04

  Ortho-condensed systems · CPC title