Phenyl glyoxal probes

Patent metadata

Field	Value
Publication number	US-9347948-B2
Application number	US-201314405467-A
Country	US
Kind code	B2
Filing date	Jun 4, 2013
Priority date	Jun 4, 2012
Publication date	May 24, 2016
Grant date	May 24, 2016

Abstract

Novel phenyl-glyoxal based anti-citrulline probes and methods of synthesis are provided. Methods of use, such as, the development of methods for monitoring substrate citrullination over time; for identifying citrullinated proteins from cells are described.

First claim

What is claimed:

1. A method of synthesizing labeled phenylglyoxal (PG) probes, comprising:
reacting an acetophenone with a bromopropionyl bromide under a nitrogen (N2) atmosphere;
precipitating a resultant product, dissolving the product in a solvent, initiating a reaction by adding sodium azide (NaN 3 ) and producing a compound having a general structure 4:

coupling a compound having a general structure 8 with a Boc-β-alanine; and, cleaving the Boc group and producing an azido-PG precursor having a general structure 9:

2. The method of claim 1 , wherein the compound having the general structure 4 is refluxed under N 2 with a selenium dioxide (SeO 2 ), producing an azido phenylglyoxal (azido-PG) having a general structure 5:

3. The method of claim 1 , wherein the compound having the general structure 9 is refluxed under N 2 with a selenium dioxide (SeO 2 ), producing an azido-PG having a general structure 10:

4. The method of claim 2 or 3 , wherein the compounds having the general structures of compounds 5 and 10 are reacted with an alkyne comprising a reporter moiety.

5. The method of claim 1 , wherein the reporter moiety comprises: fluorophores, isotopes, biotin, or rhodamine.

6. The method of claim 1 , wherein the phenylglyoxal is linked to the reporter moiety via the meta position.

7. A method of testing bioavailability of Protein Arginine Deiminase (PAD) inhibitors comprising: contacting a biological sample with an effective amount of an isotopically labeled, fluorophore labeled, or biotin labeled phenylglyoxal (biotin-PG), wherein the isotopically labeled phenylglyoxal compound is heavy phenylglyoxal (PG H ) or light phenylglyoxal (PG L ) having a general structure I:

wherein Z is 13 C for heavy phenylglyoxal (PG H ), Z is 12 C for light phenylglyoxal (PG L ) and Y is H; or, a phenylglyoxal probe produced by the method of claim 1 ; and,
assaying for the labeled proteins as compared to a control for testing the bioavailability of PAD inhibitors.

8. A method of identifying Protein Arginine Deiminase (PAD) inhibitors comprising: contacting a biological sample with an effective amount of an isotopically labeled, fluorophore labeled, or biotin labeled phenylglyoxal (biotin-PG), wherein the isotopically labeled phenylglyoxal compound is heavy phenylglyoxal (PG H ) or light phenylglyoxal (PG L ) having a general structure I:

wherein Z 13 C is for heavy phenylglyoxal (PG H ), Z is 12 C for light phenylglyoxal (PG L ) and Y is H; or, a phenylglyoxal probe produced by the method of claim 1 ; and, assaying for citrullination of proteins as compared to a control.

Assignees

• Scripps Research Inst

Inventors

• Thompson Paul R
• Bicker Kevin L
• Subramanian Venkataraman

Classifications

• C07D311/90

  with hydrocarbon radicals, substituted by amino radicals, directly attached in position 9 · CPC title

• G01N33/68

  involving proteins, peptides or amino acids {(involving lipoproteins G01N33/92)} · CPC title

• G01N2333/775

  Apolipopeptides · CPC title

• C12Q1/34 Primary

  involving hydrolase · CPC title

• G01N33/532

  Production of labelled immunochemicals · CPC title