Synthetic oligosaccharide subunits of the Psl exopolysaccharide of pseudomonas aeruginosa and uses thereof

Patent metadata

Field	Value
Publication number	US-10870710-B2
Application number	US-201816219199-A
Country	US
Kind code	B2
Filing date	Dec 13, 2018
Priority date	May 14, 2013
Publication date	Dec 22, 2020
Grant date	Dec 22, 2020

Abstract

This disclosure relates to synthetic oligosaccharide subunits of the Pseudomonas exosaccharide Psi and uses thereof, e.g., for epitope mapping of anti-Psl antibodies, for identification of anti-Psl antibodies, and for use as vaccines. In one aspect a synthetic oligosaccharide subunit of a Pseudomonas aeruginosa Psl oligosaccharide is provided, comprising the trisaccharide of formula I.

First claim

What is claimed is:

1. A method comprising

(a) reacting a thiomannosyl donor 7 with acceptor

saccharide 10 to form disaccharide 13, and then partially deprotecting disaccharide 13 to form disaccharide 14:

(b) reacting disaccharide 14 and with thiomannoside 8

to form trisaccharide 15 then partially deprotecting the trisaccharide to form compound 16,

then reacting compound 16 with CC 1 3 CN, and Cs 2 CO 3 , in dichloromethane to form the trichloroacetimidate trisaccharide 17:
and

(c) reacting the trichloroacetimidate trisaccharide 17 with compound 12

in the presence of trimethylsilyl trifluoromethanesulfonate and dichloromethane at −30° C. to form a linear tetrasaccharide 18

then partially deprotecting linear tetrasaccharide 18 with hydrofluoric acid in pyridine to form the linear tetrasaccharide 19:

and wherein Nap is 2-methylnaphthyl, Ph is a phenyl group, Bn is a benzyl group, Bz is a benzoyl group, All is allyl ether, and TBS is t-butyldimethylsilyl.

2. The method of claim 1 , further comprising deprotecting the linear tetrasaccharide 19 to form tetrasaccharide 3:

3. The method of claim 1 , further comprising
(d) reacting the linear tetrasaccharide 19 with compound 9

in the presence of trimethylsilyl trifluoromethanesulfonate and dichloromethane to form a pentasaccharide 20 then partially deprotecting

pentasaccharide 20 to form a pentasaccharide 21:

wherein Ac is an acetyl group.

4. The method of claim 3 , further comprising deprotecting the pentasaccharide 20 to form a pentasaccharide 2:

5. The method of claim 3 , further comprising
(e) reacting the pentasaccharide 21 with compound 11

in the presence of N-iodosuccinimide, trimethylsilyl trifluoromethanesulfonate and dichloromethane to form a hexasaccharide 22

then partially deprotecting the hexasaccharide 22 with hydrofluoric acid in pyridine to form a hexasaccharide 6:

wherein Et is an ethyl group.

6. The method of claim 5 , further comprising deprotecting compound the hexasaccharide 6 to form a hexasaccharide 4:

7. The method of claim 5 , further comprising:
(f) partially deprotecting the trisaccharide 15 in the presence of hydrofluoric acid

in pyridine to form a trisaccharide 23 and reacting the trisaccharide 23 with compound 9 in the presence of trimethylsilyl trifluoromethanesulfonate and dichloromethane to form a tetrasaccharide 24 then reacting the tetrasaccharide 24

in the presence of (Ph 3P ) 3 RhC1, N,N-disopropylethylamine in toluene, followed by HgO, HgC 1 2 , in

acetone/H2O to form a tetrasaccharide 25, then deprotecting the tetrasaccharide 25 with 2,2,2-trifluoro-N-phenylacetimidoyl chloride, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) in dichloromethane to form a tetrasaccharide 5:

(g) reacting hexasaccharide 6 with tetrasaccharide 5 in the presence of trimethylsilyl trifluoromethanesulfonate and dichloromethane to form a decasaccharide 26 then deprotecting decasaccharide 26 to form compound 1:

Assignees

• Medlmmune Ltd
• Univ Georgia
• Medimmune Ltd

Inventors

• Digiandomenico Antonio
• Wang Qun
• Stover Charles K
• Boons Geert-Jan
• Mo Kai-For
• Li Huiqing

Classifications

• G01N33/56911 Primary

  Bacteria · CPC title

• G01N33/5308

  for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites · CPC title

• G01N2333/21

  from Pseudomonadaceae (F) · CPC title

• C08B37/006

  Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof · CPC title

• A61P37/04

  Immunostimulants · CPC title