Labeled chiral alpha-hydroxy ketoacid derivatives, a process for preparing said derivatives and their use

Patent metadata

Field	Value
Publication number	US-9784744-B2
Application number	US-201314419759-A
Country	US
Kind code	B2
Filing date	Aug 7, 2013
Priority date	Aug 8, 2012
Publication date	Oct 10, 2017
Grant date	Oct 10, 2017

Abstract

The present invention relates to labeled chiral alpha-hydroxy ketoacid derivatives, a process for preparing the derivatives and their use for isotopic labeling of amino acids, in particular, for isotopic labeling of methyl groups of amino acids, and more particularly, for specific isotopic labeling of valine, leucine and isoleucine methyl groups, in proteins and biomolecular assemblies. The invention also concerns a process for analyzing proteins and biomolecular assemblies by NMR spectroscopy including a step of isotopic labeling of amino acids, in particular, valine, leucine and isoleucine, in proteins and biomolecular assemblies to be analyzed by the chiral alpha-hydroxy ketoacid derivatives of the invention.
The invention further relates to a kit for isotopic labeling of valine, leucine and isoleucine amino acids, in proteins and biomolecular assemblies including one or more chiral alpha-hydroxy ketoacid derivatives of the invention.

First claim

The invention claimed is:

1. A composition consisting essentially of a compound of formula (I) having an (S) configuration:

wherein
X 1 and X 2 are, independently from each other, 1 H (H) or 2 H (D);
Y 1 , Y 2 and Y 3 are, independently from each other, 12 C (C) or 13 C;
R 1 is a methyl group in which the carbon atom is 12 C (C) or 13 C and the hydrogen atoms are, independently from each other, 1 H (H) or 2 H (D);
R 2 is a methyl group in which the carbon atom is 12 C (C) or 13 C and the hydrogen atoms are, independently from each other, 1 H (H) or 2 H (D); or
R 2 is an ethyl group in which the carbon atoms are, independently from each other, 12 C (C) or 13 C and the hydrogen atoms are, independently from each other, 1 H (H) or 2 H (D),
with the proviso that in compound of formula (I), at the same time, at least one hydrogen atom is 2 H (D) and at least one carbon atom is 13 C.

2. The composition of claim 1 , wherein in the compound of formula (I)
R 1 is chosen from a group consisting of CD 3 , 13 CH 3 , 13 CH 2 D and 13 CHD 2 ;
R 2 is a methyl group chosen from a group consisting of 13 CD 3 ; or
R 2 is an ethyl group chosen from a group consisting of CD 3 -CD 2 , 13 CH 3 -CD 2 , CH 3 — 13 CD 2 , 13 CHD 2 -CD 2 , 13 CHD 2 - 13 CD 2 , 13 CH 2 D-CD 2 , 13 CH 2 D- 13 CD 2 and 13 CH 3 — 13 CD 2 .

3. The composition of claim 1 , wherein in the compound of formula (I) Y 1 ═Y 2 ═Y 3 ═C.

4. The composition of claim 1 , wherein in the compound of formula (I) Y 1 ═Y 2 ═Y 3 ═ 13 C.

5. The composition of claim 1 , wherein said compound of formula (I) is chosen from the group consisting of:
(S)-2-(1′- 2 H2, 2′- 13 C)ethyl-2-hydroxy-3-oxo-4-( 2 H 3 ) butanoic acid {X 1 ═X 2 ═H; Y 1 ═Y 2 ═Y 3 ═C; R 1 ═CD 3 ; R 2 ═ 13 CH 3 —CD 2 };
(S)-2-(1′- 2 H2, 2′- 2 H, 2′- 13 C)ethyl-2-hydroxy-3-oxo-4-( 2 H 3 )butanoic acid {X 1 ═X 2 ═H; Y 1 ═Y 2 ═Y 3 ═C; R 1 ═CD 3 ; R 2 ═ 13 CH 2 D − CD 2 };
(S)-2-(1′- 2 H 2 , 2′- 2 H 2 , 2′- 13 C,)ethyl-2-hydroxy-3-oxo-4-( 2 H 3 )butanoic acid {X 1 ═X 2 ═H; Y 1 ═Y 2 ═Y 3 ═C; R 1 ═CD 3 ; R 2 ═ 13 CHD 2− CD 2 };
(S)-2-hydroxy-2-( 2 H 3 , 13 C)methyl-3-oxo-4( 13 C)butanoic acid {X 1 ═X 2 ═H; Y 1 ═Y 2 ═Y 3 ═C; R 1 ═ 13 CH 3 ; R 2 ═ 13 CD 3 };
(S)-2-( 2 H5)ethyl-2-hydroxy-3-oxo-4-( 13 C)methylbutanoic acid {X 1 ═X 2 ═H; Y 1 ═Y 2 ═Y 3 ═C; R 1 ═ 13 CH 3 ; R 2 ═CD 3− CD 2 }.

6. The composition of claim 1 , wherein said compound of formula (I) is chosen from the group consisting of:
(S)-1,2,3-( 13 C)-2-(1′- 2 H2, 13 C2)ethyl-2-hydroxy-3-oxo-4-( 2 H3)butanoic acid {X 1 ═X 2 ═H; Y 1 ═Y 2 ═Y 3 ═ 13 C; R 1 ═CD 3 ; R 2 ═CH 3− 13 CD 2 };
(S)-1,2,3-( 13 C)-2-(1′- 2 H2, 2′- 2 H, 13 C 2 )ethyl-2-hydroxy-3-oxo-4-( 2 H3)butanoic acid {X 1 ═X 2 ═H; Y 1 ═Y 2 ═Y 3 ═ 13 C; R 1 ═CD 3 ; R 2 ═ 13 CH 2 D- 13 CD 2 };
(S)-1,2,3-( 13 C)-2-(1′- 2 H2, 2′- 2 H2, 13 C 2 )ethyl-2-hydroxy-3-oxo-4-( 2 H 3 )butanoic acid {X 1 ═X 2 ═H; Y 1 ═Y 2 ═Y 3 ═ 13 C; R 1 ═CD 3 ; R 2 ═ 13 CHD 2− 13 CD 2 }.

Assignees

• Commissariat Energie Atomique
• Centre Nat Rech Scient
• Centre Nat De La Recherch Scient

Inventors

• Plevin Michael
• Kerfah Rime
• Pessey Ombeline
• Boisbouvier Jérôme
• Gans Pierre

Classifications

• C07B59/001 Primary

  Acyclic or carbocyclic compounds · CPC title

• C07C59/215

  containing singly bound oxygen containing groups · CPC title

• G01N33/58 Primary

  involving labelled substances (G01N33/53 takes precedence) · CPC title

• G01N33/6812

  Assays for specific amino acids · CPC title

• C12P7/42

  Hydroxy-carboxylic acids · CPC title