Compositions and methods for modification of biomolecules

Patent metadata

Field	Value
Publication number	US-12048710-B2
Application number	US-202217672508-A
Country	US
Kind code	B2
Filing date	Feb 15, 2022
Priority date	Nov 1, 2004
Publication date	Jul 30, 2024
Grant date	Jul 30, 2024

Abstract

The present invention provides modified cycloalkyne compounds; and method of use of such compounds in modifying biomolecules. The present invention features a cycloaddition reaction that can be carried out under physiological conditions. In general, the invention involves reacting a modified cycloalkyne with an azide moiety on a target biomolecule, generating a covalently modified biomolecule. The selectivity of the reaction and its compatibility with aqueous environments provide for its application in vivo (e.g., on the cell surface or intracellularly) and in vitro (e.g., synthesis of peptides and other polymers, production of modified (e.g., labeled) amino acids).

First claim

What is claimed is:

1. A compound of the formula:

wherein each of R 1 -R 6 is independently selected from the group consisting of H; one or two halogen atoms; a carboxylic acid; a methoxy group; an alkyl ester; an aryl ester; a substituted aryl ester; an aldehyde; an amine; a thiol; an amide; an aryl amide; an alkyl halide; a thioester; a sulfonyl ester; an alkyl ketone; an aryl ketone; a substituted aryl ketone; a halosulfonyl; a nitrile; a nitro; —W—(CH 2 ) n —Z, wherein n is an integer from 1 to 4, W is O, N, or S, and Z is nitro, cyano, sulfonic acid, or a halogen; —(CH 2 ) n —W—(CH 2 ) m —Z, wherein n and m are each independently 1 or 2, W is O, N, S, or sulfonyl, wherein if W is O, N, or S, then Z is nitro, cyano, or halogen, and wherein if W is sulfonyl, then Z is H; and —(CH 2 ) n —Z, wherein n is an integer from 1 to 4, and Z is nitro, cyano, sulfonic acid, or a halogen; and
wherein each of R 1 -R 6 is optionally independently linked to a moiety that comprises a reactive group that facilitates covalent attachment of a molecule of interest; or a molecule of interest.

2. A compound of the formula:

where R 1 and R 2 are each independently selected from the group consisting of H; one or two halogen atoms; a carboxylic acid; an alkyl ester; an aryl ester; a substituted aryl ester; an aldehyde; an amine; a thiol; an amide; an aryl amide; an alkyl halide; a thioester; a sulfonyl ester; an alkyl ketone; an aryl ketone; a substituted aryl ketone; a halosulfonyl; a nitrile; and a nitro; and
wherein each of R 1 and R 2 is optionally independently linked to a moiety that comprises a reactive group that facilitates covalent attachment of a molecule of interest; or a molecule of interest.

3. A method for synthetically modifying a cellular component, the method comprising:
introducing an azide moiety into a cellular component, thereby generating an azide-modified cellular component; and
contacting the cell comprising the azide-modified cellular component with a reactive partner comprising a compound of claim 1 , said contacting being under physiological conditions;
wherein said contacting with said reactive partner results in reaction between the azide group of azide-modified cellular component and the compound of claim 1 , thereby synthetically and covalently modifying the cellular component.

4. The method of claim 3 , wherein cellular component is a polypeptide.

5. The method of claim 3 , wherein said contacting is in vivo.

6. A method for synthetically modifying a cellular component, the method comprising:
introducing an azide moiety into a cellular component, thereby generating an azide-modified cellular component; and
contacting the cell comprising the azide-modified cellular component with a reactive partner comprising a compound of claim 2 , said contacting being under physiological conditions;
wherein said contacting with said reactive partner results in reaction between the azide group of azide-modified cellular component and the compound of claim 2 , thereby synthetically and covalently modifying the cellular component.

7. The method of claim 6 , wherein cellular component is a polypeptide.

8. The method of claim 6 , wherein said contacting is in vivo.

Assignees

• Univ California

Inventors

• Bertozzi Carolyn Ruth
• Agard Nicholas J
• Prescher Jennifer A
• Baskin Jeremy Michael
• Sletten Ellen May

Classifications

• C07C57/26

  containing rings other than six-membered aromatic rings · CPC title

• C07C63/74

  having unsaturation outside the aromatic rings · CPC title

• C07D225/02

  not condensed with other rings · CPC title

• C07C2601/18

  with a ring being at least seven-membered · CPC title

• G01N33/6842

  Proteomic analysis of subsets of protein mixtures with reduced complexity, e.g. membrane proteins, phosphoproteins, organelle proteins · CPC title