N-containing saccharides and method for the synthesis of N-containing saccharides from amino-deoxy-disaccharides and amino-deoxy-oligosaccharides
AssigneeBioflexin AB
InventorKurt G. I. Nilsson
Life of this patent
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Abstract
Synthesis of an amino-disaccharide, amino-oligosaccharide or a derivative thereof, characterized in that a monosaccharide, a disaccharide, an oligosaccharide, a glycoside or a derivative thereof, in the presence of a glycosidase as catalyst, is reacted with an amino-deoxy-saccharide or a glycoside or derivative thereof, and that the amino-saccharide is optionally isolated from the product mixture directly or after chemical/enzymatic modification.
Claims32
A method of producing an amino-deoxy di- or oligosaccharide compound which either consists of or is a fragment or analog of a carbohydrate part in a glycoconjugate, said method comprising
- (1) reacting
- (a) at least one donor substance comprising a mono, di- or oligosaccharide, a glycoside or a derivative thereof,
- (b) at least one acceptor substance comprising an amino-deoxy mono-, di-, or oligosaccharide, or glycoside thereof, and
- (c) an E.C. group 3.2 glycosidase or a modified 3.2 glycosidase to form an amino-deoxy di- or oligosaccharide compound,
- wherein said glycosidase is a recombinant glycosidase, and
- (2) optionally isolating said amino-deoxy di- or oligosaccharide compound.
An amino-deoxy di- or oligosaccharide which either consists of or is a fragment or analog of a carbohydrate part in a glycoconjugate, having a formula I or II
- in which I is a derivatized 2-amino-2-deoxy-D-glucopyranoside and II is a derivatized 2-amino-2-deoxy-D-galactopyranoside and at least one of R1, R3, R4, and R6 is a hydroxyl group or an organic or an inorganic group and R2 is —H2, an organic group or an inorganic group and in which at least one of R3, R4, or R6 is a mono-, di-, tri- or higher oligosaccharide group which is glycosidically bound to a remaining part of I or II and a remainder of R1, R3, R4 and R6 is non-derivatized or is derivatized with one or more organic or inorganic groups,
- provided that when said donor substance is galactosyl or sialyl 2-3 galactosyl and when said acceptor substance is 2-amino-2-deoxyglucopyranosyl then a β1-4 linkage cannot contain
- or NHR, and
- provided that said donor substance is not an (amino)monosaccharide.
An amino-deoxy di- or oligosaccharide compound which either consists of or is a fragment or analog of a carbohydrate part in a glycoconjugate, represented by the formula
- D—O-(amino-)saccharide
- wherein D is a monosaccharide, disaccharide or oligosaccharide residue which is α or β O-glycosidically linked to the (amino)saccharide, said (amino)saccharide is a 2-amino-2-deoxy-glucopyranoside, 2-amino-2-deoxy-galactopyranoside, or 2-amino-2-deoxy-mannopyranoside selected from the group consisting of:
- where R3, R4, and R6 are —OH and R1 is pentenyl-, —SEt, —SPh, OEtBr, —OEtSiMe3, —OAll, —OPh, —OCH2Ph, or —OR where R is CH3(CH2)n where n is 0-12 or where R is an amino acid or peptide,
- provided that when said donor substance is galactosyl or sialyl 2-3 galactosyl and when said acceptor substance is 2-amino-2-deoxyglucopyranosyl then a β1-4 linkage cannot contain
- or NHR, and
- provided that said donor substance is not an (amino)monosaccharide.
An amino-deoxy di- or oligosaccharide compound having the formula:
- D—O-(amino-)saccharide
- wherein D is a mono-, di- or oligosaccharide group or an analog thereof, which is α- or β- or O-glycosidically bound to the -(amino-)saccharide, and wherein -(amino-)saccharide is obtained from:
- (1) an amino-deoxy mono- or di- or oligosaccharide, or glycoside thereof;
- (2) an amino-deoxy-glucopyranoside, an amino-deoxy-galactopyranoside, or an amino-deoxy-mannopyranoside;
- (3) a 2-amino-2-deoxy-saccharide substituted in the 3, 4, 5 or 6 OH group with an organic or inorganic group selected from the group consisting of allyloxy-(CH2═CH—CH2O—), benzyloxy-(PhCH2O—), benzoyloxy-(PhCOO—), chloroacetyloxy-(ClCH2COO), p-methoxybenzyloxy-(p-MeO-PhCH2O—), trityl-(Ph3CO—), trialkylsilyloxy-, tosylate-, mesylate-, phosphate-, ONO3, sulfate-, carboxylate, and RCOO— where R is CH3(CH2)n and where n is 1-20;
- or where two vicinal hydroxyl groups have been replaced with a benzylidene acetal, isopropylidene ketal or an ortho ester, pivaloyl-group, tetrahydropyranyl, (2-methoxyethoxy)methylisopropylidene ketal, cyclohexylidene ketal, benzylidene acetal, orthoester, benzoyloxy-(PhCOO—);
- or
- (4) an amino-deoxy-di, tri-, or higher oligosaccharide,
- provided that when said donor substance is galactosyl or sialyl 2-3 galactosyl and when said acceptor substance is 2-amino-2-deoxyglucopyranosyl then a β1-4 linkage cannot contain
- or NHR, and
- provided that said donor substance is not an (amino)monosaccharide.
An amino-deoxy di- or oligosaccharide compound represented by the formula
- D—O-(amino-)saccharide
- wherein D is a monosaccharide, disaccharide or oligosaccharide residue which is α or β O-glycosidically linked to the (amino)saccharide, said (amino)saccharide is a member selected from the group consisting of
- wherein one or two of the hydroxyl groups of the positions 2, 3, 4, 5 or 6 have been replaced with allyloxy-(CH2═CH—CH2O—), benzyloxy-(PhCH2O—), benzoyloxy-(PhCOO—), chloroacetyloxy-(ClCH2COO), p-methoxybenzyloxy-(p-MeO-PhCH2O—), trityl-(Ph3CO—), trialkylsilyloxy-, tosylate-, mesylate-, phosphate-, sulfate-, carboxylate, pivaloyl-group, tetrahydropyranyl, (2-methoxyethoxy)methylisopropylidene ketal, cyclohexylidene ketal, benzylidene acetal, orthoester, —ONO3, sulfate-, phosphate-, carboxylate, —OC(O)R as acetyl-, butanoyl-, octanoyl-, benzoyl, -pivaloyl, and RCOO— where R is CH3(CH2)n and where n=1-20, or a pivaloyloxy-group, or where two vicinal hydroxyl groups have been replaced with a benzylidene acetal, isopropylidene ketal or an ortho ester.
An amino-deoxy di- or oligosaccharide compound which either consists of or is a fragment or analog of the carbohydrate part in a glycoconjugate, prepared by a method comprising the steps of
- (1) reacting
- (a) at least one mono-, di- or oligosaccharide, glycoside, or derivative thereof as donor substance,
- (b) at least one acceptor substance comprising an amino-deoxy mono-, di-, oligosaccharide, glycoside or derivative thereof, and
- (c) an E.C. group 3.2 glycosidase to form said aminodeoxy di- or oligosaccharide compound, and
- (2) optionally isolating said amino-deoxy di- or oligosaccharide compound,
- provided that when said donor substance is galactosyl or sialyl 2-3 galactosyl and when said acceptor substance is 2-amino-2-deoxyglucopyranosyl then a β1-4 linkage cannot contain
- or NHR, and
- provided that said donor substance is not an (amino)monosaccharide.
A method of synthesizing an amino-disaccharide, amino-oligosaccharide or a derivative thereof, comprising reacting a monosaccharide, disaccharide, oligosaccharide, glycoside or derivative thereof in the presence of a recombinant glycosidase or a modified glycosidase as catalyst with an amino-deoxy-saccharide or derivative thereof and optionally isolating the amino-saccharide from the product mixture directly or after further chemical/enzymatic modification.
A method of producing an amino-deoxy di- or oligosaccharide compound selected from the group consisting of Galβ1-3(6-O-Bn)GlcNH2βSEt, Fucα1-4(6-O-Bn)GlcNH2βSEt, Galβ1-4(6-OBn)GlcNH2βSEt, Galβ1-3GlcNH2βSEt, Galβ1-3GalNH2βSEt, Galβ1-4GlcNH2βSEt, Galβ1-3(6-OAll)GlcNH2βSEt, Galβ1-4(6-OAll)GlcNH2βSEt, Galβ1-3(4-OBn)GlcNH2βSEt, Galβ1-4(3-OBn)GlcNH2βSEt, Fucα1-4(6-OBn)GlcNH2βSEt, Fucα1-3(6-OBn)GlcNH2βSet, Fucα1-3(4-OBn)GlcNH2βSEt, Fucα1-4(3-OBn)GlcNH2βSEt, GlcNAcβ1-3(6-OBn)GlcNH2βSEt, GlcNAcβ1-4(6-OBn)GlcNH2βSEt, GlcNAcβ1-4(3-OBn)GlcNH2βSEt, GlcNAcβ1-3(4-OBn)GlcNH2βSEt, GlcNAcβ1-3(6-OBn)GalNH2βSEt, GlcNAcβ1-4(6-OBn)GalNH2βSEt, GlcNAcβ1-4(3-OBn)GalNH2,βSEt, GlcNAcβ1-3(4-OBn)GalNH2βSEt, GalNAcβ1-3(6-OBn)GlcNH2βSEt, GalNAcβ1-4(6-OBn)GlcNH2βSEt, GalNAcβ1-4(3-OBn)GlcNH2βSEt, GalNAcβ1-3(4-OBn)GlcNH2βSEt, GalNAcβ1-3(6-OBn)GalNH2βSEt, GalNAcβ1-4(6-OBn)GalNH2βSEt, GalNAcβ1-4(3-OBn)GalNH2βSEt, GalNAcβ1-3(4-OBn)GalNH2βSEt, GalNAcα1-3(6-OBn)GlcNH2βSEt, GalNAcα1-4(6-OBn)GlcNH2βSEt, GalNAcα1-(3-OBn)GlcNH2βSEt, GalNAcα1-3(4-OBn)GlcNH2βSEt, GalNAcα1-3(6-OBn)GalNH2βSEt, GalNAcα1-4(6-OBn)GalNH2βSEt, GalNAcα1-4(3-OBn)GalNH2βSEt, GalNAcα1-3(4-OBn)GalNH2βSEt, Manα1-3(6-OBn)GlcNH2βSEt, Manα1-4(6-OBn)GlcNH2βSEt, Manα1-4(3 OBn)GlcNH2βSEt, Manα1-3(4-OBn)GlcNH2βSEt, Manα1-3(6-OBn)GalNH2βSEt, Manα1-4(6-OBn)GalNH2βSEt, Manα1-4(3-OBn)GalNH2βSEt, Manα1-3(4-OBn)GalNH2βSEt, Glcβ1-3(6-OBn)GlcNH2βSEt, Glcβ1-4(6-OBn)GlcNH2βSEt, Glcβ1-4 (3-OBn)GlcNH2βSEt, Glcβ1-3(4-OBn)GlcNH2βSEt, Glcβ1-3(6-OBn)GalNH2βSEt, Glcβ1-4(6-OBn)GalNH2βSEt, Glcβ1-4(3-OBn)GalNH2βSEt, Glcβ1-3(4-OBn)GalNH2βSEt, NeuAcα2-3Galβ1-3GlcNH2βSEt, NeuAcα2-3Galβ1-4GlcNH2βSEt, NeuAcα2-3Galβ1-4(6-OBn)GlcNH2βSEt, NeuAcα2-3Galβ1-3(4-OBn)GlcNH2βSEt, NeuAcα2-3Galβ1-4(3-OBn)GlcNH2βSEt, NeuAcα2-6Galβ1-4GlcNH2βSEt, Galα1-3Galβ1-4GlcNH2βSEt, Galβ1-4(Fucα1-3)GlcNH2βSEt, Fucα1-2Galβ1-4GlcNH2βSEt, Fucα1-2Galβ1-3GlcNH2βSEt, NeuAcα2-3Galβ1-3GalNH2βSEt, and NeuAcα2-3Galβ1-3(NeuAcα2-6)GalNH2βSEt, comprising
- (1) reacting
- (a) at least one donor substance comprising a mono-, di-, or oligosaccharide, a glycoside or a derivative thereof
- (b) at least one acceptor substance comprising an amino-deoxy mono-, di, or oligosaccharide, or glycoside thereof, and
- (c) an E.C. group 3.2 glycosidase or a modified 3.2 glycosidase to form an amino-deoxy di- or oligosaccharide, and compound, wherein said glycosidase is a recombinant glycosidase,
- (2) optionally isolating said amino-deoxy di- or oligosaccharide compound.
A method of producing an amino-deoxy di- or oligosaccharide compound selected from the group consisting of Galβ1-3(6-O-Bn)GlcNH2βSEt, Fucα1-4(6O-Bn)GlcNH2βSEt, Galβ1-4(6-OBn)GlcNH2βSEt, Galβ1-3GlcNH2βSEt, Galβ1-3GalNH2βSEt, Galβ1-4GlcNH2βSEt, Galβ1-3(6-OAll)GlcNH2βSEt, Galβ1-4(6-OAll)GlcNH2βSEt, Galβ1-3(4-OBn)GlcNH2βSEt, Galβ1-4(3-OBn)GlcNH2βSEt, Fucα1-4(6-OBn)GlcNH2βSEt, Fucα1-3(6-OBn)GlcNH2βSet, Fucα1-3(4-OBn)GlcNH2βSEt, Fucα1-4(3-OBn)GlcNH2βSEt, GlcNAcβ1-3(6-OBn)GlcNH2βSEt, GlcNAcβ1-4(6-OBn)GlcNH2βSEt, GlcNAcβ1-4(3-OBn)GlcNH2βSEt, GlcNAcβ1-3(4-OBn)GlcNH2βSEt, GlcNAcβ1-3(6-OBn)GalNH2βSEt, GlcNAcβ1-4(6-OBn)GalNH2βSEt, GlcNAcβ1-4(3-OBn)GalNH2,βSEt, GlcNAcβ1-3(4-OBn)GalNH2βSEt, GalNAcβ1-3(6-OBn)GlcNH2βSEt, GalNAcβ1-4(6-OBn)GlcNH2βSEt, GalNAcβ1-4(3-OBn)GlcNH2βSEt, GalNAcβ1-3(4-OBn)GlcNH2βSEt, GalNAcβ1-3(6-OBn)GalNH2βSEt, GalNAcβ1-4(6-OBn)GalNH2βSEt, GalNAcβ1-4(3-OBn)GalNH2βSEt, GalNAcβ1-3(4-OBn)GalNH2βSEt, GalNAcα1-3(6-OBn)GlcNH2βSEt, GalNAcα1-4(6-OBn)GlcNH2βSEt, GalNAcα1-(3-OBn)GlcNH2βSEt, GalNAcα1-3(4-OBn)GlcNH2βSEt, GalNAcα1-3(6-OBn)GalNH2βSEt, GalNAcα1-4(6-OBn)GalNH2βSEt, GalNAcα1-4(3-OBn)GalNH2βSEt, GalNAcα1-3(4-OBn)GalNH2βSEt, Manα1-3(6-OBn)GlcNH2βSEt, Manα1-4(6-OBn)GlcNH2βSEt, Manα1-4(3 OBn)GlcNH2βSEt, Manα1-3(4-OBn)GlcNH2βSEt, Manα1-3(6-OBn)GalNH2βSEt, Manα1-4(6-OBn)GalNH2βSEt, Manα1-4(3-OBn)GalNH2βSEt, Manα1-3(4-OBn)GalNH2βSEt, Glcβ1-3(6-OBn)GlcNH2βSEt, Glcβ1-4(6-OBn)GlcNH2βSEt, Glcβ1-4 (3-OBn)GlcNH2βSEt, Glcβ1-3(4-OBn)GlcNH2βSEt, Glcβ1-3(6-OBn)GalNH2βSEt, Glcβ1-4(6-OBn)GalNH2βSEt, Glcβ1-4(3-OBn)GalNH2βSEt, Glcβ1-3(4-OBn)GalNH2βSEt, NeuAcα2-3Galβ1-3GlcNH2βSEt, NeuAcα2-3Galβ1-4GlcNH2βSEt, NeuAcα2-3Galβ1-4(6-OBn)GlcNH2βSEt, NeuAcα2-3Galβ1-3(4-OBn)GlcNH2βSEt, NeuAcα2-3Galβ1-4(3-OBn)GlcNH2βSEt, NeuAcα2-6Galβ1-4GlcNH2βSEt, Galα1-3Galβ1-4GlcNH2βSEt, Galβ1-4(Fucα1-3)GlcNH2βSEt, Fucα1-2Galβ1-4GlcNH2βSEt, Fucα1-2Galβ1-3GlcNH2βSEt, NeuAcα2-3Galβ1-3GalNH2βSEt, and NeuAcα2-3Galβ1-3(NeuAcα2-6)GalNH2βSEt,
- (1) reacting
- (a) at least one mono-, di- or oligosaccharide, glycoside, or derivative thereof as donor substance,
- (b) at least one acceptor substance comprising an amino-deoxy mono-, di-, oligosaccharide, glycoside or derivative thereof, and
- (c) an E.C. group 3.2 glycosidase to form said aminodeoxy di- or oligosaccharide compound, and
- (2) optionally isolating said amino-deoxy di- or oligosaccharide compound.
An amino-deoxy di- or oligosaccharide compound which either consists of or is a fragment or analog of a carbohydrate part in a glycoconjugate, produced by a method comprising
- (1) reacting
- (a) at least one donor substance comprising a mono-, di- or oligosaccharide, a glycoside or a derivative thereof,
- (b) at least one acceptor substance comprising an amino-deoxy mono-, di-, or oligosaccharide, or glycoside thereof, and
- (c) an E.C. group 3.2 glycosidase or a modified 3.2 glycosidase to form an amino-deoxy di- or oligosaccharide compound,
- (2) optionally isolating said amino-deoxy di- or oligosaccharide compound,
- provided that when said donor substance is galactosyl or sialyl 2-3 galactosyl and when said acceptor substance is 2-amino-2-deoxyglucopyranosyl then a β1-4 linkage cannot contain
- or NHR, and
- provided that said donor substance is not an (amino)monosaccharide.
Dependencies are inferred from the claim text.