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US 7,598,372 B2Granted
Synthesis of core sugar chain structure of asparagine-linked glycoprotein
AssigneeShionogi & Co., Ltd.
InventorsShinichiro Nishimura, Yasuhiro Takegawa
Life of this patent
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Abstract
It is intended to chemically synthesize the trisaccharide moiety at the reducing end in the core sugar chain structure of an asparagine-linked glycoprotein. By using a highly inexpensive natural polysaccharide having a mannoseβ-1,4-bond as the starting material, a β-1,4-glycoside bond of mannose is formed.
Claims9
A method for preparing a trisaccharide (Manβ1→4GlcNβ1→4GlcN) of a reducing terminal in the core sugar chain structure of an asparagine-linked glycoprotein, comprising
- (1) a process of preparing a mannose disaccharide compound (a type of ManP1β1→4ManP1) of the formula (I)
- wherein P1, OH-protecting group selected from the group consisting of acetyl, benzyl, 4-methoxybenzyl, benzoyl, methoxymethyl, tetrahydropyranyl, trimethylsilyl and triethylsilyl, and the wavy line means that —OP1 is linked at an axial or equatorial position, or mixture of both, by hydrolyzing a polysaccharide having mannoseβ-1,4-bonds and protecting OH groups of the resulting hydrolysate,
- (2) a process of preparing a glycal compound, in which mannose of a reducing terminal of the mannose disaccharide is converted to glycal, by halogenation and reduction of the mannose disaccharide (a type of ManP1β1→4ManP1),
- (3) a process of preparing an azide disaccharide compound (a type of ManP1β1→4ManP1) shown with formula (II) in which a 2-azide group of mannose in a reducing terminal is linked at an equatorial position;
- wherein P1 is the same as described above, the wavy line means that —ONO2 is linked at an axial or equatorial position, or mixture of both,
- by azidenitration reaction of the glycal compound above,
- (4) a process of substituting the nitro group of the azide disaccharide compound (a type of ManP1β1→4ManP1) with a leaving group selected from the group consisting of fluorine atom, chlorine atom, trihaloacetoimidate, 4-pentenyl, alkylthio and arylthio, and
- (5) a process of preparing a trisaccharide compound (a type of Manβ1→4GlcNP1β1→4GlcNP2) shown with the formula (III);
- wherein P1 is an OH-protecting group, as described above, P2 is an OH-protecting group selected from the group consisting of acetyl, benzyl, 4-methoxybenzyl, benzoyl, methoxymethyl, tetrahydropyranyl, trimethylsilyl and triethylsilyl, P3 is an amino-protecting group selected from the group consisting of phthalimide, tert-butyloxycarbonyl, benzyloxycarbonyl, acetyl, benzoyl and benzyl, and P11 is an OH-protecting group selected from the group consisting of acetyl, benzyl, 4-methoxybenzyl, benzoyl, methoxymethyl, tetrahydropyranyl, trimethylsilyl and triethylsilyl,
- by a reaction of the product having the leaving group with amino-protected glucopyranoside shown with the formula;
- wherein P2, P3 and P11 are the same as described above.
8 more elements
A method for preparing the azide disaccharide (a type of ManP1β1→4ManP1) shown with the formula (II) in which the 2-azide group of mannose in a reducing terminal is linked at the equatorial position;
- wherein P1 is an OH-protecting group selected from the group consisting of acetyl, benzyl, 4-methoxybenzyl, benzoyl, methoxymethyl, tetrahydropyranyl, trimethylsilyl and triethylsilyl, and the wavy line means that —ONO2 is linked at an axial or equatorial position, or mixture of both, comprising a process of preparing a glycal compound, in which mannose of the reducing terminal of the mannose disaccharide is converted to glycal, by halogenation and reduction of the mannose disaccharide compound (a type of ManP1β1→4ManP1) shown with the formula (I);
- wherein P1 is the same as described above and the wavy line means that —OP1 is linked at an axial or equatorial position, or mixture of both,
- and subsequent azidenitration reaction of the glycal compound.
A method for preparing the trisaccharide compound shown with the formula (III);
- wherein P1, P2, and P11 are independently OH-protecting groups selected from the group consisting of acetyl, benzyl, 4-methoxybenzyl, benzoyl, methoxymethyl, tetrahydropyranyl, trimethylsilyl and triethylsilyl, and P3 is an amino-protecting group selected from the group consisting of phthalimide, tert-butyloxycarbonyl, benzyloxycarbonyl, acetyl, benzoyl and benzyl, comprising a process of substituting the nitro group of the azide disaccharide compound (a type of ManP1β1→4ManP1) shown with the formula (II) with a leaving group selected from the group consisting of fluorine atom, chlorine atom, trihaloacetoimidate, 4-pentenyl, alkylthio and arylthio;
- wherein P1 is the same as described above, the wavy line means that —ONO2 is linked at an axial or equatorial position, or mixture of both, and a 2-azide group of mannose in the reducing terminal is linked at the equatorial position,
- and next, reacting the substituted compound having the leaving group with amino-protected glucopyranoside of the formula;
- wherein P2, P3 and P11 are the same as described above.
1 more element
A method for preparing an asparagine-linked trisaccharide compound (Manβ1→4GlcNP1β1→4GlcNP2) shown with the formula (IV)
- wherein P1 and P2 are independently OH-protecting groups selected from the group consisting of acetyl, benzyl, 4-methoxybenzyl, benzoyl, methoxymethyl, tetrahydropyranyl, trimethylsilyl and triethylsilyl, P4 and P6 are independently amino-protecting groups selected from the group consisting of phthalimide, tert-butyloxycarbonyl, benzyloxycarbonyl, acetyl, benzoyl and benzyl, and P5 is a carboxyl-protecting group which is t-Bu,
- by deprotecting the P11 group of the compound (III),
- wherein P1 and P2 are the same as described above, P3 is an amino-protecting group selected from the group consisting of phthalimide, tert-butyloxycarbonyl, benzyloxycarbonyl, acetyl, benzoyl and benzyl, and P11 is an OH-protecting group selected from the group consisting of acetyl, benzyl, 4-methoxybenzyl, benzoyl, methoxymethyl, tetrahydropyranyl, trimethylsilyl and triethylsilyl,
- reducing the azide group to an amino group, protecting the amino group with an acetyl group, forming an oxazoline ring simultaneously with deprotecting a hydroxy group of a reducing terminal, and coupling with a protected aspartic acid derivative of the formula:
- wherein P5 and P6 are the same as described above,
- after introducing a —N═C═S group at the reducing terminal.
3 more elements
The azide disaccharide (a type of ManP1β1→4ManP1) compound shown with the formula (II);
- wherein P1 is an OH-protecting group selected from the group consisting of acetyl, benzyl, 4-methoxybenzyl, benzoyl, methoxymethyl, tetrahydropyranyl, trimethylsilyl and triethylsilyl, and the wavy line means that —ONO2 is linked at an axial or equatorial position, or mixture of both.
A trisaccharide compound (a type of Manβ1→4GlcNP1β1→4GlcNP2) shown with the formula of (III);
- wherein P1, P2 and P11 are independently OH-protecting groups selected from the group consisting of acetyl, benzyl, 4-methoxybenzyl, benzoyl, methoxymethyl, tetrahydropyranyl, trimethylsilyl and triethylsilyl, and P3 is an amino-protecting group selected from the group consisting of phthalimide, tert-butyloxycarbonyl, benzyloxycarbonyl, acetyl, benzoyl and benzyl.
A method for preparing a trisaccharide (Manβ1→4GlcNβ1→4GlcN) of a reducing terminal in a core sugar chain structure of an asparagine-linked glycoprotein, comprising
- (1) a process of preparing a mannose disaccharide compound (a type of ManP1β1→4ManP1) of the formula (I)
- wherein P1 is an OH-protecting group selected from the group consisting of acetyl, benzyl, 4-methoxybenzyl, benzoyl, methoxymethyl, tetrahydropyranyl, trimethylsilyl and triethylsilyl, and the wavy line means that —OP1 is linked at an axial or equatorial position, or mixture of both, by hydrolyzing guar gum or galactomannan of the formula (V);
- wherein n is an integer of 50 or more,
- and protecting OH groups of the resulting hydrolysate.
- (2) a process of preparing a glycal compound, in which mannose of a reducing terminal of the mannose disaccharide is converted to glycal, by halogenation and reduction of the mannose disaccharide (a type of ManP1β1→4ManP1), and
- (3) a process of preparing an azide disaccharide compound (a type of ManP1β1→4ManP1) shown with formula (II) in which a 2-azide group of mannose in a reducing terminal is linked at an equatorial position;
- wherein P1 is the same as described above, the wavy line means that —ONO2 is linked at an axial or equatorial position, or mixture of both,
- by azidenitration reaction of the glycal compound above,
- (4) a process of substituting the nitro group of the azide disaccharide compound (a type of ManP1β1→4ManP1) with a leaving group selected from the group consisting of fluorine atom, chlorine atom, trihaloacetoimidate, 4-pentenyl, alkylthio and arylthio, and
- (5) a process of preparing a trisaccharide compound (a type of Manβ1→4GlcNP1β1→4GlcNP2) shown with the formula (III);
- wherein P1 an OH-protecting group, as described above, P2 is an OH-protecting group selected from the group consisting of acetyl, benzyl, 4-methoxybenzyl, benzoyl, methoxymethyl, tetrahydropyranyl, trimethylsilyl and triethylsilyl, P3 is an amino-protecting group selected from the group consisting of phthalimide, tert-butyloxycarbonyl, benzyloxycarbonyl, acetyl, benzoyl and benzyl, and P11 is an OH-protecting group selected from the group consisting of acetyl, benzyl, 4-methoxybenzyl, benzoyl, methoxymethyl, tetrahydropyranyl, trimethylsilyl and triethylsilyl,
- by a reaction of the product having the leaving group with amino-protected glucopyranoside shown with the formula;
- wherein P2, P3, and P11 are the same as described above.
10 more elements
Dependencies are inferred from the claim text.