Fluorescent detection of amines and hydrazines and assaying methods thereof

Patent metadata

Field	Value
Publication number	US-11530352-B2
Application number	US-201716316996-A
Country	US
Kind code	B2
Filing date	Oct 2, 2017
Priority date	Oct 3, 2016
Publication date	Dec 20, 2022
Grant date	Dec 20, 2022

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 assaying a compound of R—NH 2 , comprising:
forming a compound of Formula (I):

by reacting a compound of Formula (III):

with the compound of R—NH 2 in the presence of a cyanide ion; and
detecting and measuring the fluorescence of the compound of Formula (I),

wherein:
X is CR a or N,
R 1 and R 2 are Ra; 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 Ra 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 ; and
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—NH 2 occurs under a mildly basic condition having a pH of from 7.5 to 9.5.

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

3. The process of claim 1 , wherein R is optionally substituted pyrimidyl, optionally substituted triazinyl, optionally substituted purinyl, or optionally substituted phenyl, each of which is attached to a solid support directly or via a linker.

4. The process of claim 1 , wherein R—NH 2 is an optionally substituted nucleobase, which is attached to a solid support directly or via a linker.

5. The process of claim 4 , wherein the nucleobase is cytosine, guanine, adenine, thymine, 7-methylguanine, 7-methylguanosine, 5-methylcytosine, 5-hydroxymethylcytosine, or 5-methylcytidine, which is attached to a solid support directly or via a linker.

6. The process of claim 4 , wherein the nucleobase is attached to a sugar moiety, which is further attached to a solid support directly or via a linker.

7. The process of claim 1 , wherein the —NH 2 group in R—NH 2 has a pKa of from about 7 to about 10.

8. The process of claim 1 , wherein R is a gel or a linker that is attached to a gel.

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

where [SS] indicates a solid support.

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

11. The process of claim 1 , wherein the cyanide ion for the preparation of the compound of Formula (I) is provided from a cyanide source of potassium cyanide or sodium cyanide.

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

13. 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.

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

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

16. The process or method of claim 1 , wherein each instance of R a1 is independently halogen, alkyl, hydroxyl, alkoxy, cyano, amido, carboxyl, acyl, acyloxy, or alkoxycarbonyl.

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

18. 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.

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

20. The process of claim 1 , wherein R—NH 2 is attached to a solid support, and together with the solid support and optional linker comprises: [SS]—OSi(OZ) 2 -alkyl-NH 2 , wherein [SS] is a solid support, and each Z is alkyl, H, or a silicon atom of an adjacent solid support-bound group.

Assignees

• Illumina Inc

Inventors

• Asadi Ali
• Li jian-sen

Classifications

• C07D209/62 Primary

  Naphtho [c] pyrroles; Hydrogenated naphtho [c] pyrroles · CPC title

• G07C3/14

  Quality control systems · CPC title

• C07D237/26

  condensed with carbocyclic rings or ring systems · CPC title

• G01N31/22

  using chemical indicators (G01N31/02 takes precedence) · CPC title

• G01N2021/3155

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