# US 10,870,710 B2 · Synthetic oligosaccharide subunits of the Psl exopolysaccharide of pseudomonas aeruginosa and uses thereof

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- Number: US 10,870,710 B2
- Document ID: US10870710B2
- Status: Granted
- Kind: B2
- Application number: 16/219,199
- Filed: 2018-12-13
- Published: 2019-10-10
- Granted: 2020-12-22
- Earliest claimed: 2013-05-14
- Assignees: MedImmune, LLC; University of Georgia Research Foundation, Inc.
- Applicants: Medlmmune Limited; University Of Georgia Research Foundation, Inc.
- Inventors: Antonio DiGiandomenico; Qun Wang; Charles K. Stover; Geert-Jan Boons; Kai-For Mo; Huiqing Li
- Web page: https://patentgrep.com/patent/US10870710B2

## CPC

- C08B 37/0006: {Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid}
- A61K 39/104: {Pseudomonadales, e.g.} Pseudomonas
- A61K 47/643: {Albumins, e.g. HSA, BSA, ovalbumin or a Keyhole Limpet Hemocyanin [KHL]}
- A61K 47/646: {the entire peptide or protein drug conjugate elicits an immune response, e.g. conjugate vaccines}
- A61P 31/04: Antibacterial agents
- A61P 37/04: Immunostimulants
- C07H 15/04: attached to an oxygen atom of the saccharide radical
- C08B 37/006: {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}
- G01N 33/5308: {for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites}
- G01N 33/56911: {Bacteria}
- A61K 2039/6081: {Albumin; Keyhole limpet haemocyanin [KLH]}
- G01N 2333/21: from Pseudomonadaceae (F)
- G01N 2400/00: Assays, e.g. immunoassays or enzyme assays, involving carbohydrates
- G01N 2400/02: involving antibodies to sugar part of glycoproteins
- G01N 2400/38: Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence, e.g. gluco- or galactomannans, Konjac gum, Locust bean gum or Guar gum
- G01N 2500/04: Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)
- C08B: POLYSACCHARIDES; DERIVATIVES THEREOF (polysaccharides containing less than six saccharide radicals attached to each other by glycosidic linkages C07H; fermentation or enzyme-using processes C12P19/00; sugar industry C13; production of cellulose D21)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## 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.

## Claims

### Claim 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 CC13 CN, and Cs2 CO3, 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.

### Claim 2 (depends on 1)

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

### Claim 3 (depends on 1)

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.

### Claim 4 (depends on 3)

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

### Claim 5 (depends on 3)

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.

### Claim 6 (depends on 5)

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

### Claim 7 (depends on 5)

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 (Ph3P)3 RhC1, N,N-disopropylethylamine in toluene, followed by HgO, HgC12, 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: