Synthesis of sialylated/fucosylated human milk oligosaccharides

Patent metadata

Field	Value
Publication number	US-10005807-B2
Application number	US-201414783783-A
Country	US
Kind code	B2
Filing date	Apr 11, 2014
Priority date	Apr 12, 2013
Publication date	Jun 26, 2018
Grant date	Jun 26, 2018

Abstract

An achemo-enzymatic synthesis of oligosaccharides of formula 1 is presented wherein R is selected from —OH, —N 3 and —OR 6 wherein R 6 is selected from allyl optionally substituted by one or more methyl, propargyl optionally substituted by one or more methyl, 2-trimethylsilyl-ethyl, —(CH 2) n —NH 2 and —(CH 2) n —N 3 wherein integer n is to 10, preferably 2 or 3, R is selected from sialyl moiety, —SO 3 H and —CH(R)—COOH wherein R is selected from H, alkyl and benzyl, R 2 is selected from H and fucosyl, R 3 is selected from H and sialyl, R 4 is selected from H and fucosyl, provided that at least one of R 3 and R 4 is H, and A is a divalent carbohydrate linker, having important biological activities and significant commercial value for the pharmaceutical and food industry.

First claim

The invention claimed is:

1. A method for making a 3-O-galactosyl-GlcNAc or -GalNAc derivative of formula 1 and salts thereof

wherein
R is selected from the group consisting of —OH, —N 3 , and —OR 6 , wherein R 6 is selected from the group consisting of allyl optionally substituted by one or more methyl, propargyl optionally substituted by one or more methyl, 2-trimethylsilyl-ethyl, —(CH 2 ) n —NH 2 , and —(CH 2 ) n —N 3 , wherein integer n is 1 to 10;
R 1 is a sialyl moiety or —CH(R 5 )—COOH, wherein R 5 is selected from the group consisting of H, alkyl, and benzyl;
R 2 is H or a fucosyl moiety;
R 3 is H or a sialyl moiety;
R 4 is H or a fucosyl moiety, provided that at least one of R 3 and R 4 is H; and
A is a divalent carbohydrate linker;
comprising the steps:
a) sialylation or addition of a —HC(R 5 )-COOH group of a compound of formula 2

wherein
R′ is —N 3 or —OR′ 6 , wherein R′ 6 is selected from the group consisting of allyl optionally substituted by one or more methyl, propargyl optionally substituted by one or more methyl, a group removable by hydrogenolysis, 2-trimethylsilyl-ethyl, and —(CH 2 ) n —N 3 ,
wherein integer n is 1 to 10;
R 7 is independently acyl;
Y is selected from the group consisting of —NHAc, haloalkanoylamido, —NAc 2 , haloalkoxycarbonylamino, 2,3-diphenylmaleimido, 2,3-dimethylmaleimido, and —N 3 ; and
B is divalent carbohydrate linker in protected form;
b) optional fucosylation of the compound obtained in step a);
c) de-O-acylation and/or basic hydrolysis, optional mild acidic hydrolysis and optional transformation of Y to —NHAc of the compound obtained in step a) or step b);
d) optional sialylation or fucosylation of the compound obtained in step c); and
e) optional catalytic hydrogenolysis and/or anomeric deprotection of the compound obtained in step d).

2. The method according to claim 1 , wherein sialylation or addition of a —HC(R 5 )-COOH group of the compound of formula 2 in step a) results in the formation of a compound of formula 4

wherein
R′ is —N 3 or —OR′ 6 , wherein R′ 6 is selected from the group consisting of allyl optionally substituted by one or more methyl, propargyl optionally substituted by one or more methyl, a group removable by hydrogenolysis, 2-trimethylsilyl-ethyl, and —(CH 2 ) n —N 3 , wherein integer n is 1 to 10;
R 7 is independently acyl;
Y is selected from the group consisting of —NHAc, haloalkanoylamido, —NAc 2 , haloalkoxycarbonylamino, 2,3-diphenylmaleimido, 2,3-dimethylmaleimido, and —N 3 ;
B is divalent carbohydrate linker in protected form; and
P is a protected sialyl moiety or —CH(R 5 )—COOR 10 , wherein R 5 is selected from the group consisting of H, alkyl, and benzyl, and R 10 is alkyl or benzyl.

3. The method according claim 2 , wherein the sialylation in step a) is carried out by reacting the compound of formula 2 with a compound of formula 3

wherein
R 8 is acyl;
Q is alkyl or benzyl; and
R 9 is optionally substituted phenyl or benzyl;
to give a compound of formula 4A

wherein
R′ is —N 3 or —OR′ 6 , wherein R′ 6 is selected from the group consisting of allyl optionally substituted by one or more methyl, propargyl optionally substituted by one or more methyl, a group removable by hydrogenolysis, 2-trimethylsilyl-ethyl, and —(CH 2 ) n —N 3 , wherein integer n is 1 to 10;
R 7 is independently acyl;
Y is selected from the group consisting of —NHAc, haloalkanoylamido, —NAc 2 , haloalkoxycarbonylamino, 2,3-diphenylmaleimido, 2,3-dimethylmaleimido, and —N 3 .

4. The method according to claim 3 , wherein the compound of formula 4A is a compound of formula 4E

wherein
R″ is a group removable by hydrogenolysis;
Y is selected from the group consisting of —NHAc, haloalkanoylamido, —NAc 2 , haloalkoxycarbonylamino, 2,3-diphenylmaleimido, 2,3-dimethylmaleimido, and azido;
R 7 is independently acyl;
R 8 is independently acyl;
R 20 is independently acyl;
R 21 is H or acyl; and
Q is alkyl or benzyl.

5. The method according to claim 1 , wherein the optional fucosylation in step b) comprises the reaction of a compound of formula 4

wherein
R′ is —N 3 or —OR′ 6 , wherein R′ 6 is selected from the group consisting of allyl optionally substituted by one or more methyl, propargyl optionally substituted by one or more methyl, a group removable by hydrogenolysis, 2-trimethylsilyl-ethyl, and —(CH 2 ) n —N 3 , wherein integer n is 1 to 10;
R 7 is independently acyl;
Y is selected from the group consisting of —NHAc, haloalkanoylamido, —NAc 2 , haloalkoxycarbonylamino, 2,3-diphenylmaleimido, 2,3-dimethylmaleimido, and —N 3 ;
B is divalent carbohydrate linker in protected form; and
P is a protected sialyl moiety or —CH(R 5 )—COOR 10 , wherein R 5 is selected from the group consisting of H, alkyl, and benzyl, and R 10 is alkyl or benzyl;
with a compound of formula 5

wherein
X is selected from the group consisting of a halogen, —OC(═NH)CCl 3 , —O-pentenyl, —OAc, —OBz, and —SR 13 , wherein R 13 is alkyl or optionally substituted phenyl;
R 11 is acyl or a group removable by hydrogenolysis; and
R 12 is a group removable by hydrogenolysis, acyl, or two R 12 groups together that form a moiety

wherein R 14 and R 15 independently are alkyl or phenyl, or wherein R 14 and R 15 together with the carbon atom, to which they are attached, form a cycloalkylidene;
to give a compound of formula 6

6. The method according to claim 5 , wherein the compound of formula 4 is a compound of formula 4E

wherein
R″ is a group removable by hydrogenolysis;
Y is selected from the group consisting of —NHAc, haloalkanoylamido, —NAc 2 , haloalkoxycarbonylamino, 2,3-diphenylmaleimido, 2,3-dimethylmaleimido, and azido;
R 7 is independently acyl;
R 8 is independently acyl;
R 20 is independently acyl;
R 21 is H or acyl; and
Q is alkyl or benzyl; and
the compound of formula 5 is a compound of formula 5A

wherein
X A is alkylthio or optionally substituted phenylthio;
R 22 and R 23 are, independently, a group removable by hydrogeno

Assignees

• Glycom As

Inventors

• Chassagne Pierre
• Khanzhin Nikolay
• Hederos Markus Jondelius
• Champion Elise
• Matwiejuk Martin
• Dekany Gyula

Classifications

• C07H5/04 Primary

  to nitrogen · CPC title

• C07H1/00

  Processes for the preparation of sugar derivatives · CPC title