JP2001089494A - Sugar chain compound and its use - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a sugar chain compound easily bondable with a compound having a group non-reactive with carboxyl group as exclusive group such as protein, enzyme, carrier and pigment without causing steric hindrance, etc., in introduction and keeping the reactivity with an antibody even after the introduction as an antigen. SOLUTION: The present invention relates to a sugar chain compound having an optionally substituted amino group bonded through a bivalent hydrocarbon residue, a carrier holding the sugar chain compound in immobilized state, a human serum antigen sensitized particle immobilized with a sugar chain compound containing an antigen site same as the human blood type antigen among the above sugar chain compounds, a reagent for the determination of human blood type containing the sugar chain compound and a method for determining human blood type by using the sugar chain compound as an antigen.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a sugar chain compound in which an optionally substituted amino group is bonded to a sugar chain having antigenicity via a divalent hydrocarbon residue, and uses thereof. It is about.

[0002]

2. Description of the Related Art In recent years, with the development of sugar chain engineering, it has been found that complex sugar chain compounds have biological significance. For example, the fact that blood group antigen substances such as ABO blood group substances and Lewis blood group substances present on the surface of cells in blood are sugar chains having a specific structure, and by immunological analysis using monoclonal antibodies, It has been revealed that cancer antigen-related substances are sugar chain compounds that have undergone specific chemical modification. In addition, glycolipids, which are a kind of sugar chain compounds, not only function as antigens but also function as cell adhesion molecules and are involved in a wide range of biological phenomena such as cell development, blood cell differentiation, and immune cell differentiation. It has been revealed.

[0003] Through these studies, sugar chain compounds have attracted attention not only as cancer-related research reagents or judgment reagents but also as detection materials for various biological components including blood group antigens.

At present, as the sugar chain compound, natural products (purified and extracted from a biological sample or the like), semi-synthetic products, total synthetic products, etc. are used, or an appropriate protein,
It is used in a state of being directly or indirectly supported by an enzyme, a carrier or the like via a spacer or the like.

[0005] However, in the determination of the ABO blood group back test, there are various problems when red blood cells having a specific human blood group antigen are used as conventional antigens.
In other words, since human erythrocytes are used as an antigen-carrying carrier, if human erythrocytes are infected with a virus, a so-called biohazard problem may occur, and human erythrocytes may not have sufficient stability as a reagent. That is it.

Therefore, in order to solve the above-mentioned problems, erythrocyte-derived sugar chain antigens or synthetic sugar chain antigens are carried on a carrier such as latex, yeast cells or zeolite particles by physical adsorption and used instead of erythrocytes. Studies have been carried out to avoid the effects of viruses (J. Ogiwara, General Clinical Studies 14 , 445 (1965), Yukio Minashiki, 27th Annual Meeting of the Japanese Society of Transfusion Society, Japanese Patent Application Laid-Open No. 60-40958) Etc.), the antigen obtained by such a loading method by physical adsorption has a problem in stability because the antigen is easily dissociated from the carrier.

As a means for solving this problem, an attempt has been made to carry an antigen on a carrier by utilizing a chemical bond by a reactive functional group (JP-A-63-241357).
Examples of such a sugar chain compound into which a reactive functional group is introduced include, for example, sugar chain derivatives (USP4137401, USP4238473) into which a carboxyl group, an alkoxycarbonyl group or the like is introduced.
Etc.), but those having a reactive functional group of a carboxyl group type have an amino group, a thiol group,
Although it can be directly reacted and supported as long as it has a hydroxyl group, an isocyanate group or a halogenated alkyl group, the carrier is, for example, a carboxyl group,
Alkoxycarbonyl group, acyloxycarbonyl group,
Haloformyl group, chlorosulfonyl group, cyanate group,
When only having a group that does not react with a carboxyl group such as an epoxy group, a ketone group, and an aldehyde group,
A complicated operation such as reacting a diamine or the like to introduce an amino group and then binding the amino group is required.

However, even when an amino group is introduced by such a method, the distance between the amino group and the sugar chain as an antigen is not so long, so that introduction into a carrier is hindered by steric hindrance, or liposome When used as a carrier, problems such as inability to maintain antigen activity after introduction may occur. Furthermore, in a sugar chain derivative having a carboxyl group derived from a sialic acid residue or the like which is an important constituent factor such as α-fetoprotein or Lewis antigen in the molecule, selective protection of the carboxyl group which requires multiple steps is required. Otherwise, there is a problem that introduction of a reactive functional group and reaction with a protein, enzyme, carrier or dye cannot be selectively performed.

[0009]

DISCLOSURE OF THE INVENTION The present invention has been made in view of the above situation, and has been developed in such a manner that a compound having only a group which does not react with a carboxyl group, such as a protein, an enzyme, a carrier, a dye, etc. It is an object of the present invention to provide a sugar chain compound which can be easily bound without causing any obstacle or the like, and which can maintain the reactivity with an antibody as an antigen after introduction.

[0010]

The present invention comprises the following constitution. (1) A sugar chain compound in which an amino group which may have a substituent is bonded via a divalent hydrocarbon residue. (Hereinafter, it may be abbreviated as the sugar chain compound of the present invention.) (2) The sugar chain comprises the same antigen site as the human blood group antigen
The sugar chain compound according to (1). (3) General formula [1]

[0011]

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(Wherein R ¹ is a hydroxyl group or —NHCOCH ₃
Wherein R ² represents an amino group which may have a substituent, X represents a bond or a sugar residue, and E represents a divalent hydrocarbon residue. The sugar chain compound according to (1), which is represented by (1). (4) A carrier on which the sugar chain compound according to (1) is immobilized. (5) Human serum antigen-sensitized particles to which the sugar chain compound according to (2) or the sugar chain compound according to (3) is immobilized. (6) A reagent for determining a human blood group comprising the sugar chain compound according to (2) or the sugar chain compound according to (3). (7) A method for determining a human blood group, comprising using the sugar chain compound according to (2) or the sugar chain compound according to (3) as an antigen.

That is, the present inventors have developed a sugar chain compound which does not have a problem of biohazard, is stable as a reagent, and can be easily chemically bonded to a carrier having only a group which does not react with a carboxyl group. As a result of intensive research,
The present inventors have found that a sugar chain compound in which an amino group which may have a substituent is bonded via a divalent hydrocarbon residue has all of the above-mentioned features, and completed the present invention. .

The sugar chain compound of the present invention is, for example, represented by the following general formula [2]

[0015]

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(Wherein E represents a divalent hydrocarbon residue,
R ² represents an amino group which may have a substituent. ) And an oxygen atom of a sugar chain compound composed of two or more monosaccharide residues.

In the general formula [2], the divalent hydrocarbon residue represented by E is usually a divalent hydrocarbon residue having 1 to 20, preferably 2 to 10, more preferably 3 to 8 carbon atoms. Aliphatic hydrocarbon residue, divalent alicyclic hydrocarbon residue, divalent alicyclic aliphatic hydrocarbon residue, divalent aromatic residue, divalent araliphatic residue, etc. These divalent hydrocarbon residues may have, for example, 1 to 3 hetero atoms such as an oxygen atom, a sulfur atom and a nitrogen atom in the chain.

The divalent aliphatic hydrocarbon residue includes, for example, a saturated aliphatic hydrocarbon residue and an unsaturated aliphatic hydrocarbon residue.

The saturated aliphatic hydrocarbon residue may be linear or branched. Specifically, for example, a methylene group,
Ethylene, trimethylene, propylene, tetramethylene, 2-methylpropylene, pentamethylene, 2-methylbutylene, 2-ethylpropylene,
Hexamethylene, 2-ethylbutylene, heptamethylene, 2-ethylpentyl, 2-methylhexylene, octamethylene, 2-ethylhexylene, nonamethylene, decamethylene, undecamethylene, dodeca An alkylene group such as a methylene group, a tetradecamethylene group, a hexadecamethylene group, an octadecamethylene group, a 2-ethyloctadecene group, an icocene group, and the like, for example, an oxygen atom, a sulfur atom, a nitrogen atom, and the like in a chain of the alkylene group -CH ₂ comprising a heteroatom e.g. _{-S-CH 2 CH 2 -,} - C
_{H 2 CH 2 -O-CH 2} CH 2 -, - CH 2 -NH-CH 2 CH (CH 3) CH 2 - and the like.

Examples of the unsaturated aliphatic hydrocarbon residue include those having 1 to 3 double bonds in the chain of the above-mentioned saturated aliphatic hydrocarbon residue. Or branched, and specifically, for example, -CH = CH-, -CH ₂ CH =
CH-, -CH ₂ -O-CH = CH _2- , -CH = CH-CH _2- , -CH ₂ CH ₂ CH = CH-,-
CH ₂ C (CH ₃ ) = CH-, -CH (CH ₂ CH ₃ ) -CH ₂ CH ₂ CH = CH-, -CH ₂ CH ₂ CH
= CH-CH = CH -, - CH 2 CH = CH-CH (CH 3) -CH = CH-CH 2 - , and the like.

The divalent alicyclic hydrocarbon residue may be monocyclic or polycyclic, and may have a double bond. Specifically, for example, a cyclopropylene group, 1,3 -Cyclopentylene group, 1,4-cyclohexylene group, 1,2-cyclohexylene group, 1,3-cyclohexylene group, 3-cyclohexene-1,2-ylene group, 2,5-cyclohexadiene-1, 4-ylene group, 1,4-cycloheptylene group, 1,4-cyclooctylene group, 2,7-spiro [3,4] octylene group, 3,9-spiro [4,
5] Deca-1,6-dienylene group,

[0022]

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And the like.

As the divalent alicyclic aliphatic hydrocarbon residue,
A group having an alicyclic hydrocarbon residue as described above in the chain or at the terminal of the saturated or unsaturated aliphatic hydrocarbon residue as described above, specifically, for example,

[0025]

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And the like.

The divalent aromatic residue may be monocyclic or polycyclic, or may have a bond between the rings. Specifically, for example, an o-phenylene group, p- -Phenylene group, m-phenylene group, 1,5-naphthylene group, 2,6-naphthylene group, biphenyl-4,4'-ylene group, 1,5-anthrylene group, 2,6-anthrylene group, p-ter Phenyl-4,4 "-ylene group, m-terphenyl-4,4'-ylene group, 2,7-phenanthrylene group, 1,6-pyrenylene group and the like.

As the divalent araliphatic residue, 1 to 3 divalent aromatic residues as described above are present in the chain or at the terminal of the above-mentioned saturated or unsaturated aliphatic hydrocarbon residue. And specifically, for example,

[0029]

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And the like.

Examples of the substituent of the optionally substituted amino group represented by R ² include a hydrogen atom from an aliphatic ring of an alkyl group, a haloalkyl group, an aryl group or a condensed polycyclic hydrocarbon. Monovalent group, arylalkyl group, acyl group, halogenated acyl group, alkoxycarbonyl group, halogenated alkoxycarbonyl group, aryloxycarbonyl group, arylalkyloxycarbonyl group, condensed polycyclic hydrocarbon fat Alkyloxycarbonyl groups and acyloxy groups having a monovalent group formed by removing one hydrogen atom from an aromatic ring as a substituent, for example, an alkylene group, a halogenated alkylene group, an arylene group, a diacyl group, a halogenated diacyl group, an alkylenedioxy group Carbonyl group, arylenedioxycarbonyl group, diacyloxy group, etc. Bruno two hydrogen atoms simultaneously substituted can divalent radical group, an amino acid residue, such as peptide residue, and the like. Incidentally, when the amino group has two substituents,
The substituents may be the same or different.

When the sugar chain compound of the present invention is used as an antigen for judging human blood group, the amino group may be, for example, an alkyl group, a haloalkyl group, an aryl group,
A monovalent group formed by removing one hydrogen atom from an aliphatic ring of a condensed polycyclic hydrocarbon, an arylalkyl group, an acyl group, a halogenated acyl group, an alkoxycarbonyl group, a halogenated alkoxycarbonyl group, an aryloxycarbonyl group, 1 formed by removing one hydrogen atom from an arylalkyloxycarbonyl group or an aliphatic ring of a condensed polycyclic hydrocarbon
Alkyloxycarbonyl group and acyloxy group having a valence group as a substituent, for example, alkylene group, halogenated alkylene group, arylene group, diacyl group, halogenated diacyl group, alkylenedioxycarbonyl group, arylenedioxycarbonyl group, diacyloxy group A divalent group capable of simultaneously substituting two hydrogen atoms of an isoamino group is preferred. When the sugar chain compound of the present invention is used as an immunogen for producing antibodies, the amino group substituent may be
An acyl group derived from a fatty acid, an amino acid residue, a peptide residue, and the like are preferable. Further, when the sugar chain compound of the present invention is used as a raw material for preparing a liposome, a substituent of the amino group may be an acyl group derived from a fatty acid. And the like.

The alkyl group may be straight-chain, branched or cyclic, and usually has 1 to 20 carbon atoms, preferably 1 to 10 carbon atoms. Specific examples include a methyl group and an ethyl group. , N-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, tert-butyl group, n-pentyl group, isopentyl group, sec-pentyl group,
tert-pentyl group, neopentyl group, 2-methylbutyl group, 1-ethylpropyl group, 2-ethylpropyl group, n-hexyl group, isohexyl group, n-heptyl group, n-octyl group, n-nonyl group, n- Decyl group, n-dodecyl group, n-tetradecyl group, n-pentadecyl group, n-heptadecyl group, n-nonadecyl group, n-icosyl group, cyclopropyl group, cyclopentyl group, cyclohexyl group, cyclopentadecyl group, cyclononadecyl group And the like.

Examples of the haloalkyl group include those in which one or more hydrogen atoms of the above-mentioned alkyl groups are substituted with halogen atoms such as chlorine, fluorine and bromine.
Specifically, for example, chloromethyl group, dichloromethyl group, trichloromethyl group, trifluoromethyl group, dibromomethyl group, 1,1,2,2-tetrachloroethyl group, 4,4,4,
4-trifluorobutyl group, perfluoro n-pentyl group,
Examples include a perfluoro n-hexyl group, a 10,10,10-trichlorodecyl group, a perfluoro n-nonadecyl group, and a perfluoro n-icosyl group.

The alkylene group is a divalent alkyl group having 2 to 20 carbon atoms, preferably 2 carbon atoms.
Groups such as ethylene group, trimethylene group, propylene group, tetramethylene group, 2-methylpropylene group, pentamethylene group, 2-methylbutylene group, 2-ethylpropylene group, hexamethylene group, −
Ethylbutylene group, heptamethylene group, 2-ethylpentyl group, 2-methylhexylene group, octamethylene group,
2-ethylhexylene group, nonamethylene group, decamethylene group, undecamethylene group, dodecamethylene group, tetradecamethylene group, hexadecamethylene group, octadecamethylene group, 2-ethyloctadecene group, icocene group and the like. .

As the halogenated alkylene group, one or more hydrogen atoms of the alkylene group as described above are, for example, chlorine,
Examples thereof include those substituted with halogen atoms such as fluorine and bromine, and specifically, for example, chloromethylene group, dibromomethylene group, 1,2-dichloroethylene group, 2- (2,2,2-trichloroethyl)- Examples include a hexylene group, a perfluorodecamethylene group, a 2- (1,1,2,2,2-pentafluoroethyl) -octadecene group, and a perfluoroicosene group.

The aryl group may be monocyclic or polycyclic, and may have the above-mentioned alkyl group or nitro group as a substituent. Specifically, for example, a phenyl group, a p-nitrophenyl group , O-tolyl group, m-tolyl group, p-
Tolyl group, 4-nitro-o-tolyl group, 4-nitro-m-tolyl group, 2,3-xylyl group, 3,5-xylyl group, 5-nitro-2,3-
Xylyl group, mesityl group, m-cumenyl group, 1-naphthyl group, 2-naphthyl group, 1-anthryl group, 2-anthryl group,
1-phenanthryl group, 2-phenanthryl group, 3-phenanthryl group, 4-phenanthryl group, 9-phenanthryl group,
And a 1-pyrenyl group.

Examples of the monovalent group formed by removing one hydrogen atom from the aliphatic ring of the condensed polycyclic hydrocarbon include, for example, those having 8 carbon atoms.
To 20 and specifically include, for example, a 1-indenyl group, a 2-indenyl group, a 1-acenaphthenyl group, a 9-fluorenyl group, and a 1-phenalenyl group.

Examples of the arylalkyl group include those in which one or more hydrogen atoms of the above-mentioned alkyl group have been substituted with the above-mentioned aryl group. Specific examples include a benzyl group, a benzhydryl group, a phenethyl group, And a trityl group.

Examples of the arylene group include those in which an aryl group as described above is a divalent group. Specific examples include an o-phenylene group, an m-phenylene group, a p-phenylene group, and an o-phenylene group. 3-tolylene group, m-2-tolylene group, m-4-tolylene group, m-5-tolylene group, p-2-tolylene group, o-3,4-xylylene group, o-3,6-xylylene group , O-4,5-xylylene group, p-
2,3-xylylene group, p-2,6-xylylene group, mesitylene group, o-3-cumenylene group, o-4-cumenylene group, m-5-cumenylene group, p-cumenylene group, 2,3-naphthylene Group, 1,4-naphthylene group, 2,7-naphthylene group, 2,6-naphthylene group, 1,8-
Naphthylene group, 1,5-naphthylene group, 1,8-anthrylene group, 2,7-anthrylene group, 2,6-anthrylene group, 1,5-anthrylene group, 1,4-anthrylene group, 2,3-anthrylene Groups, 2,7-phenanthrylene group, 3,6-phenanthrylene group, 4,5-phenanthrylene group, 9,10-phenanthrylene group and the like.

As the acyl group, a monocarboxylic acid having 2 to 24 carbon atoms such as an acetyl group, a propionyl group,
Butyryl, isobutyryl, valeryl, isovaleryl, pivaloyl, heptanoyl, octanoyl, nonanoyl, decanoyl, lauroyl, tridecanoyl, myristoyl, pentadecanoyl,
Acyl groups such as palmitoyl, heptadecanoyl, stearoyl, nonadecanoyl, icosanoyl, henicosanoyl, docosanoyl, tricosanoyl, tetracosanoyl, benzoyl, toluoyl, hydroatropoyl, and 2-naphthoyl; Can be

When the sugar chain compound of the present invention is used as an immunogen for producing antibodies or a raw material for preparing liposomes,
Examples of the acyl group derived from a fatty acid, which is mentioned as a substituent of the amino group, include, for example, those having 12 to 24 carbon atoms, preferably 14 carbon atoms.
To 20; specifically, for example, lauroyl group, tridecanoyl group, myristoyl group, pentadecanoyl group, palmitoyl group, heptadecanoyl group, stearoyl group, nonadecanoyl group, icosanoyl group, henicosanoyl group, docosanoyl group, tricosanoyl group Group, tetracosanoyl group and the like.

Examples of the diacyl group include a dicarboxylic acid having 2 to 20 carbon atoms such as a malonyl group, a succinyl group, a glutaryl group, an adipoyl group, a heptane dioil group, an octane oil group, a nonane oil group, a decane oil group, and phthaloyl. Group, isophthaloyl group, terephthaloyl group and the like.

Examples of the acyl halide group include those in which one or more hydrogen atoms of the above-mentioned acyl group are substituted with a halogen atom such as chlorine, fluorine, and bromine. Acetyl, trichloroacetyl, tribromoacetyl, 3,3,3-trichloropropionyl, perfluoropropionyl,
3,3,4,4-tetrachlorobutyryl group, perfluorooctanoyl group, 9,9,10,10,10-pentachlorodecanoyl group,
Examples include a perbromomyristoyl group and a perfluorostearoyl group.

Examples of the diacyl halide group include those in which one or more hydrogen atoms of the diacyl group are substituted by halogen atoms such as chlorine, fluorine and bromine. 2-dichloromalonyl group, 2,3-
Examples thereof include a dibromosuccinyl group, a 4,4-difluoroheptanedioyl group, and a perfluorodecanedioil group.

Examples of the alkoxycarbonyl group include those having 2 to 11 carbon atoms. Specific examples include a methoxycarbonyl group, an ethoxycarbonyl group, a propyloxycarbonyl group, an n-butoxycarbonyl group and a sec-
Butoxycarbonyl group, tert-butoxycarbonyl group, n
-Pentyloxycarbonyl group, n-hexyloxycarbonyl group, n-heptyloxycarbonyl group, 2-ethylhexyloxycarbonyl group, n-octyloxycarbonyl group, n-nonyloxycarbonyl group, n-decyloxycarbonyl group and the like. Can be

Examples of the halogenated alkoxycarbonyl group include those in which one or more hydrogen atoms of the above-mentioned alkoxycarbonyl group are substituted with halogen atoms such as chlorine, fluorine and bromine. Methoxycarbonyl group, 2,2,2-trichloroethoxycarbonyl group, perfluoropropyloxycarbonyl group, perfluorobutoxycarbonyl group, 5,5,5-trichloropentyloxycarbonyl group, 8,8,8-trifluorooctyloxy Examples include a carbonyl group and a perfluorodecyloxycarbonyl group.

Examples of the alkylenedioxycarbonyl group include those having 3 to 12 carbon atoms. Specific examples include a methylenedioxycarbonyl group, an ethylenedioxycarbonyl group, a trimethylenedioxycarbonyl group, and a propylenedioxycarbonyl group. Oxycarbonyl group, tetramethylenedioxycarbonyl group, 3-methylpropylenedioxycarbonyl group, 2-methylbutylenedioxycarbonyl group,
2,3-dimethylpropylenedioxycarbonyl group, 3-ethylpropylenedioxycarbonyl group, hexamethylenedioxycarbonyl group, heptamethylenedioxycarbonyl group, octamethylenedioxycarbonyl group, nonamethylenedioxycarbonyl group, decamethylene And a dioxycarbonyl group.

The aryloxycarbonyl group includes, for example, those having 6 to 20 carbon atoms. Specifically, for example, phenyloxycarbonyl, o-tolyloxycarbonyl, m-tolyloxycarbonyl, p-tolyl Oxycarbonyl group, 2,3-xylyloxycarbonyl group, 2,
4-xylyloxycarbonyl group, 3,5-xylyloxycarbonyl group, 1-naphthyloxycarbonyl group, 2-naphthyloxycarbonyl group, 1-anthryloxycarbonyl group, 2-anthryloxycarbonyl group, 1- Examples thereof include a phenanthryloxycarbonyl group, a 2-phenanthryloxycarbonyl group, a 3-phenanthryloxycarbonyl group, a 4-phenanthryloxycarbonyl group, and a 9-phenanthryloxycarbonyl group.

Examples of the arylalkyloxycarbonyl group include those in which one or more hydrogen atoms of the above-described alkyloxycarbonyl group have been substituted with the above-described aryl group. Specific examples include a benzyloxycarbonyl group. , P-nitrobenzyloxycarbonyl group, benzhydryloxycarbonyl group, phenethyloxycarbonyl group, trityloxycarbonyl group and the like.

The alkyloxycarbonyl group having as a substituent a monovalent group formed by removing one hydrogen atom from the aliphatic ring of the condensed polycyclic hydrocarbon includes one or more hydrogen atoms of the alkyloxycarbonyl group as described above. Examples include those in which an atom is substituted with a monovalent group formed by removing one hydrogen atom from the aliphatic ring of the above-mentioned condensed polycyclic hydrocarbon, and specifically, for example, a 1-indenylmethyloxycarbonyl group , 2-
Examples include an indenylethyloxycarbonyl group, a 1-acenaphthenylethyloxycarbonyl group, a 1-phenalenylmethyloxycarbonyl group, and a 9-fluorenylmethyloxycarbonyl group.

The amino acid residue means a group obtained by removing the -OH group from the terminal carboxyl group of the amino acid, and specifically includes, for example, an alanine residue, an arginine residue, an asparagine residue, an aspartic acid residue, a cysteine residue. Residue, cystine residue, glutamic acid residue, glutamine residue, glycine residue, histidine residue, isoleucine residue, leucine residue, n-leucine residue, lysine residue, methionine residue, phenylalanine residue, proline Residues, serine residues, threonine residues, tryptophan residues, tyrosine residues, valine residues and the like.

The peptide residue may be linear or branched, and usually includes 2 to 100, preferably 2 to 50 amino acid residues.

As the monosaccharide residue constituting the sugar chain, 1 to 5 hydrogen atoms of the terminal OH group, preferably 1 to 5
Tetroth residues such as threose residue, erythrose residue, erythrulose residue, etc., for example, ribose residue, deoxyribose residue, arabinose residue, xylose residue, lyxose residue. , Ribulose residue, xylulose residue, arabinulose residue, pentose residues such as lixulose, for example, glucose residue, mannose residue, allose residue, altrose residue, galactose residue, talose residue, idose residue , Growth residue, fucose residue, fructose residue,
Hexose residues such as sorbose residues, tagatose residues, and psicose residues, for example, glucosamine residues, galaclosamine residues, N-acetylglucosamine residues, N-acetylgalactosamine residues, N-acetylmuramic acid residues, and the like. Amino sugar residues, for example, sialic acid residues such as N-acetylinominic acid residue, for example, sorbitol residue, inositol residue, mannitol residue, ribitol residue, erythritol residue, arabitol residue, xylitol These monosaccharide residues may have one or more of -OH in the form of -OR ', and may have two or more monosaccharide residues. Two -OH may be linked together to form -OR "-O-.

As R ′, any one can be used as long as it can replace a hydrogen atom of a hydroxyl group of a sugar (for example, one used as a protecting group for a hydroxyl group). For example, an alkyl group, an aryl group and a substituent can be used. Benzyl group, acyl group, acyl halide group, acylalkylcarbonyl group, benzylidene group, isopropylidene group, trimethylsilyl group, tert-butyldimethylsilyl group, -CH (CF ₃ ) -NHCO-O- _{CH 2 C 6 H 5, -CO} -O-CH 2 C 6 H 4 (2Br) , and the like.

R "represents, for example, a methylene group, an ethylene group, a trimethylene group, a propylene group, a tetramethylene group, a 2-methylpropylene group, a pentamethylene group,
Methylbutylene group, 2-ethylpropylene group, hexamethylene group, 2-ethylbutylene group, heptamethylene group,
2-ethylpentyl group, 2-methylhexylene group, octamethylene group, 2-ethylhexylene group, nonamethylene group, decamethylene group, undecamethylene group, dodecamethylene group, tetradecamethylene group, hexadecamethylene group, octamethylene group It usually has 1 to 20 carbon atoms, preferably 1 carbon atom such as a decamethylene group, a 2-ethyloctadecene group and an icosene group.
10 to 10 linear or branched alkylene groups such as phenylmethylene, phenylethylene, 1-phenyltrimethylene, 2-phenyltrimethylene, 1-phenyltetramethylene, and 2-phenyltetramethylene Base,
1,4-diphenyltetramethylene group, 1-phenylpentamethylene group, 2-phenylpentamethylene group, 3-phenylpentamethylene group, 1,5-diphenylpentamethylene group, benzylmethylene group, benzylethylene group, 1-benzyl Trimethylene group, 2-benzyltrimethylene group, 1-benzyltetramethylene group, 1-benzyl-4-phenyltetramethylene group, 2-benzyltetramethylene group, 1-benzylpentamethylene group, 1-benzyl-5-phenyl Examples thereof include groups in which one or more hydrogen atoms of the above alkylene group are substituted with an aromatic ring, such as a pentamethylene group, a 2-benzylpentamethylene group, and a 3-benzylpentamethylene group.

The alkyl group may be linear, branched, or cyclic, and may have, for example, 1 to 3 hetero atoms such as an oxygen atom, a sulfur atom, and a nitrogen atom. Having 1 to 10 carbon atoms such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec- Pentyl group, tert-pentyl group, neopentyl group, n-hexyl group, isohexyl group,
Cyclohexyl group,

[0058]

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And n-heptyl and cycloheptyl groups.

The aryl group may be monocyclic or polycyclic, and may have the above-mentioned alkyl group or nitro group as a substituent. Specifically, for example, a phenyl group, a p-nitrophenyl group , 2,4-nitrophenyl group, o-tolyl group, m-tolyl group, p-tolyl group, 4-nitro-o-tolyl group, 4-nitro-m-tolyl group, 2,3-xylyl group, 3 , 5-xylyl group, 5-nitro-2,3-xylyl group, mesityl group, m-cumenyl group, 1-naphthyl group, 2-naphthyl group, 1-anthryl group, 2-anthryl group, 1-phenanthryl group, 2-phenanthryl group, 3-phenanthryl group, 4-phenanthryl group,
Examples thereof include a 9-phenanthryl group and a 1-pyrenyl group.

Examples of the benzyl group which may have a substituent include, for example, a nitrobenzyl group, -CH ₂ C ₆ H ₄ (2NO ₂ ), -CH ₂
C ₆ H ₃ (2,6Cl ₂ ), —CH ₂ C ₆ H ₄ (3Br), an azidobenzyl group, a methoxybenzyl group and the like.

The acyl group includes a monocarboxylic acid having 2 to 24 carbon atoms such as an acetyl group, a propionyl group,
Butyryl, isobutyryl, valeryl, isovaleryl, pivaloyl, heptanoyl, octanoyl, nonanoyl, decanoyl, lauroyl, tridecanoyl, myristoyl, pentadecanoyl,
Examples include a palmitoyl group, a heptadecanoyl group, a stearoyl group, a nonadecanoyl group, an icosanoyl group, a henicosanoyl group, a docosanoyl group, a tricosanoyl group, a tetracosanoyl group, a benzoyl group, a toluoyl group, a hydroatropoyl group, and a 2-naphthoyl group.

Examples of the acyl halide group include those in which one or more hydrogen atoms of the above-mentioned acyl group have been substituted with a halogen atom such as chlorine, fluorine and bromine. Acetyl, trichloroacetyl, tribromoacetyl, 3,3,3-trichloropropionyl, perfluoropropionyl,
3,3,4,4-tetrachlorobutyryl group, perfluorooctanoyl group, 9,9,10,10,10-pentachlorodecanoyl group,
Examples include a perbromomyristoyl group and a perfluorostearoyl group.

Examples of the acylalkylcarbonyl group include acetylmethylcarbonyl, acetylethylcarbonyl, acetyltrimethylcarbonyl, acetylpropylcarbonyl, propionylmethylcarbonyl, propionylethylcarbonyl, propionylpropylcarbonyl, and butyrylmethylcarbonyl. Group, butyrylethylcarbonyl group and the like.

The sugar chains of the sugar chain compound of the present invention may be the same or different, usually two or more, preferably 2-8.
And more preferably 3 to 6 of the above-described monosaccharide residues bonded to each other by O-glycosidic bonds.

Specific examples of the sugar chain compound of the present invention include, for example,

[0067]

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[0068]

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[0069]

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[0070]

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[0071]

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[0072]

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[0073]

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[0074]

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[0075]

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[0076]

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[0077]

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[0078]

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[0079]

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[0080]

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[0081]

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[0082]

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[0083]

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(However, Ac represents an acetyl group, Ph represents a phenyl group, Bn represents a benzyl group, and Bz represents a benzoyl group.).

When the sugar chain compound of the present invention comprises the same antigen site as the human blood group antigen, the sugar chain compound of the present invention
For example, the general formula [1]

[0086]

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(Wherein, R ¹ , R ² , X and E are the same as described above).

In the general formula [1], one or more, preferably 1 to 5, more preferably 1 to 3, sugar residues represented by X in the present invention may be the same or different. Each of the above-described monosaccharide residues constituting the sugar chain compound is represented by O-
It is formed by bonding through glycosidic bonds.

Specific examples of the compound represented by the above general formula [1] include, for example,

[0090]

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[0091]

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[0092]

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[0093]

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[0094]

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[0095]

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(Wherein, Ac represents an acetyl group).

The sugar chain compound of the present invention in which an amino group which may have a substituent is bonded via a divalent hydrocarbon residue is, for example, a compound represented by the general formula [3]

[0098]

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(Wherein E and R ² are the same as those described above) and 2 to 8 monosaccharide residues described above, and a hydroxyl group [—OR '(R' is the same as above). And a sugar chain compound having the above formula] is subjected to an O-glycosylation reaction.

The sugar chain compound of the present invention comprises a compound represented by the general formula [3] and one or more but less than eight monosaccharide residues described above and having a hydroxyl group at an arbitrary position. After the O-glycosylation reaction with the chain compound, an appropriate number of monosaccharides may be further bound to the obtained sugar chain compound one by one by O-glycosidic bonds.

R ² is, for example, an alkyl group, a haloalkyl group, an aryl group, a monovalent group formed by removing one hydrogen atom from an aliphatic ring of a condensed polycyclic hydrocarbon, an arylalkyl group, an acyl group, an acyl halide, Group, an alkoxycarbonyl group, a halogenated alkoxycarbonyl group, an aryloxycarbonyl group, an arylalkyloxycarbonyl group, and one hydrogen atom from the aliphatic ring of a condensed polycyclic hydrocarbon.
An alkyloxycarbonyl group or an acyloxy group having a monovalent group capable of being removed as a substituent, for example, an alkylene group, a halogenated alkylene group, an arylene group, a diacyl group, a halogenated diacyl group, an alkylenedioxycarbonyl group, an arylenedioxycarbonyl group; In the case of an amino group having a substituent such as a divalent group capable of simultaneously substituting two hydrogen atoms of an amino group such as a group and a diacyloxy group, if this is treated by a conventional method, for example, R ² is an amino group The corresponding compound is easily obtained. Further, among the compounds of the present invention, those in which the hydroxyl group is -OR 'or -OR "-O- are treated by a conventional method to obtain -OR' or -O-.
The corresponding sugar chain compound in which R "-O- has become -OH can be easily obtained.

The above O-glycosylation reaction may be performed according to a conventional method. That is, the O-glycosidic bond between the sugar residue and the hydroxyl group of the compound represented by the general formula [3] may be performed, for example, as follows.

That is, a so-called sugar donor which becomes a sugar compound by the removal of a substituent such as a halogenated sugar compound such as a 1-chloro-substituted product or a 1-bromo-substituted product, and 1 to 5 equivalents of a phthalimide group,
A compound represented by the general formula [3] having an amino group protected by a suitable protecting group such as a trifluoroacetyl group;
Dissolve in dichloromethane, nitromethane, acetonitrile, etc., and add 1 to 5 equivalents of an activator such as silver perchlorate and silver carbonate, silver trifluoromethanesulfonate and s-collidine or mercury bromide and mercury cyanide. Preferably, the reaction is carried out with stirring at -30 ° C. to room temperature for 2 to 24 hours in the presence of a dehydrating agent such as molecular sieves 4A or dry alite, and then purified by a silica gel column or the like to obtain the desired compound. Can be.

When a 1-thioalkylated sugar such as a 1-thiomethyl-substituted product, a 1-thioethyl-substituted product, or a 1-thiophenyl-substituted product is used as the sugar donor, 1 to 5 equivalents of the sugar donor are used. Is dissolved in dichloromethane, nitromethane, acetonitrile or the like with a compound represented by the general formula [3] having an amino group protected with a suitable protecting group such as a phthalimide group or a trifluoroacetyl group, and the solution is dissolved in 1 to 5 equivalents. ,
For example, an activator such as methyl triflate, copper bromide and tetrabutylammonium bromide or N-succinimide and trifluoromethanesulfonic acid is added, and preferably coexistence of a dehydrating agent such as molecular sieves 4A or dryalite. Under a temperature of -30 ° C to room temperature for 2 to 24 hours with stirring,
Then, by purifying with a silica gel column or the like, the target compound can be obtained.

Further, in order to form an appropriate sugar chain and a sugar chain obtained by O-glycosidically bonding the compound represented by the general formula [3] to the reducing end, a glycoside bond may be carried out as follows, for example. .

That is, as a sugar donor, a 1-chloro substituent,
When a halogenated sugar such as a 1-bromo substituent is used, 1 to 5 equivalents of a hydroxyl group other than a hydroxyl group to form an O-glycosidic bond with a sugar donor are converted to a sugar such as an acetyl group, a benzoyl group or a benzyl group. Protected with a protecting group commonly used in the synthesis of
In addition, a sugar chain compound in which an amino group bonded via a divalent hydrocarbon residue is protected with a suitable protecting group such as a phthalimide group or a trifluoroacetyl group is dissolved in dichloromethane, nitromethane, acetonitrile, or the like. 55 equivalents of an activator such as silver perchlorate and silver carbonate, silver trifluoromethanesulfonate and s-collidine or mercury bromide and mercury cyanide are added, preferably such as molecular sieves 4A or drylite. In the presence of a dehydrating agent, at -30 ° C to room temperature, 2
The target compound can be obtained by reacting with stirring for 2424 hours, and then purifying with a silica gel column or the like.

Alternatively, 1-thiomethyl, 1-thioethyl, 1-thiophenyl, etc.
When a thioalkylated sugar is used, 1 to 5 equivalents of a hydroxyl group other than a hydroxyl group to form an O-glycoside bond with a sugar donor are usually used in the synthesis of a sugar such as an acetyl group, a benzoyl group or a benzyl group. A sugar chain compound in which an amino group protected by a protecting group and bound via a divalent hydrocarbon residue is protected by a suitable protecting group such as a phthalimide group or a trifluoroacetyl group is treated with dichloromethane, nitromethane, or acetonitrile. And an activator such as 1 to 5 equivalents, for example, methyl triflate, copper bromide and tetrabutylammonium bromide or N-succinimide and trifluoromethanesulfonic acid, and preferably molecular -30 ℃ in the presence of a dehydrating agent such as Sieves 4A or Dryalite
The reaction is carried out under stirring at room temperature to room temperature for 2 to 24 hours, and then purification is performed using a silica gel column or the like, whereby the target compound can be obtained.

The monosaccharide used for producing the sugar chain compound of the present invention may be the same as the monosaccharide which is the group of the monosaccharide residue constituting the sugar chain compound of the present invention. A hydrogen atom directly bonded to the main chain of
It may be substituted by N ₃ or the like.

The hydroxyl group which may be possessed by the monosaccharide has a part or all of -OR '(R' is the same as described above), for example,-
It may be OCNHCCl ₃ , 2- (trimethylsilyl) ethoxy group or the like.

According to the present invention, via a divalent hydrocarbon residue,
A sugar chain compound to which an amino group which may have a substituent is bonded has an amino group which may have a substituent, for example, a carboxyl group, an alkoxycarbonyl group,
Groups derived from carboxylic acid derivatives such as alkyloxycarbonyl group, haloformyl group, chlorosulfonyl group, halogenated alkyl group, cyanate group, isocyanate group,
It can be easily bonded to a carrier or a dye having an epoxy group, a ketone group, an aldehyde group or the like.

When the sugar chain compound of the present invention has antigenicity, the sugar chain compound of the present invention can be used as an immunogen by incorporating it into a suitable carrier such as liposome, cell, peptide, fatty acid and the like. Therefore, it can be used for producing the antibody.

Among the sugar chain compounds of the present invention, those in which a fatty acid has been introduced into the amino group are useful as a raw material for producing liposomes. A liposome having the sugar chain compound of the present invention immobilized thereon can be easily obtained.

When the sugar chain compound of the present invention has an antigenic site, it is used as an antigen. For example, immunoturbidimetry (Clinical test method outline revised 30th edition, Kanbara Publishing Co., Ltd. Izumi Kanai) Original author p853), Radioimmunoassay (Clinical test method outline revised 30th edition) Kanbara Publishing Co., Ltd. Kanai Izumihara p856
Enzyme-linked immunosorbent assay (see p862 written by Kanbara Publishing Co., Ltd., Izuhara Kanai, p.862)
Immunochromatography method, fluorescence polarization method
30th edition, Kinbara Shuppan Co., Ltd., by Izuhara Kanai, p865), and the like, to confirm the presence of an antibody against the sugar chain compound.

That is, when the sugar chain compound of the present invention has, for example, the same structure as the human A-type antigen or human B-type antigen, the blood type is determined using the various methods described above. You can also.

When the blood group is determined by using the sugar chain compound of the present invention as a blood group antigen, for example, using a sensitized particle agglutination method or immunochromatography, the following method may be used.

<Sensitizing Particle Aggregation Method> When a carrier carrying the sugar chain compound of the present invention having the same antigenic site as the type A antigen is brought into contact with a biological sample such as serum or plasma, When the sample has an anti-A antibody, that is, in the case of blood types B and O, aggregation occurs, which can be confirmed visually, but when the biological sample does not have the anti-A antibody, That is, in the case of blood types A and AB, aggregation does not occur.

Conversely, when a biological sample is added to a carrier carrying the sugar chain compound of the present invention having the same antigenic site as type B antigen, the biological sample does not have an anti-B antibody. Does not agglutinate, but does occur when it has an anti-B antibody.

Further, if the above agglutination reaction is utilized, blood group A
B-type and O-type can also be easily determined. That is, a biologically-derived sample is provided for each of the carrier carrying the sugar chain compound of the present invention having the same antigen site as the type A antigen and the carrier carrying the sugar chain compound of the present invention having the same antigen site as the type B antigen. Is added, and when these both aggregate, the biological sample is O 2
If the sample is found to be of type AB and no agglutination is observed, the biological sample is found to be of type AB.

Table 1 shows combinations of antigens on erythrocytes and antibodies in serum for a given human blood group.

[0120]

[Table 1]

<Immunochromatography> The sugar chain compound of the present invention having the same antigen site as the type A (type B) antigen bound to a labeling substance (hereinafter abbreviated as labeled synthetic blood type antigen).
Is supported on an absorbent carrier so as to be movable by capillary action, and the sugar chain compound of the present invention having the same antigen site as the A-type (B-type) antigen in another place (hereinafter referred to as synthetic blood group) An abbreviated antigen) is supplied to a portion of the absorbent carrier on which the labeled synthetic blood group antigen is retained, and a biological sample is provided on the resulting synthetic blood group antigen-supporting portion.
The blood type of the biological sample is determined based on the color development derived from the labeling substance.

The biological sample used in the sensitized particle aggregation method and the immunochromatography method may be any sample having an anti-A antibody and / or an anti-B antibody.

In the sensitized particle agglutination method, any carrier having a functional group which reacts with an amino group may be used as a carrier for supporting a sugar chain compound, and may be a carrier used in a usual immunoassay. There are, for example, polystyrene, polypropylene, polyamide, polyvinyl alcohol, polyvinyl acetate, polyester, polyacrylic acid,
Polyvinyl chloride, polyvinylidene chloride, polyethylene,
Synthetic polymer compounds such as polychlorocarbonate, silicone resin, and silicone rubber, such as porous glass, ground glass, alumina, silica gel, activated carbon, and inorganic substances such as metal oxides react with amino groups, such as carboxyl groups, carbonyls Examples thereof include those into which functional groups such as groups have been introduced, and these carriers are used in the form of particles such as fine particles and latex particles. Among these, latex particles derived from a synthetic polymer compound are preferably used from the viewpoint that the carrier surface can be easily chemically treated according to the purpose and that a non-specific reaction hardly occurs.

Also, proteins such as liposomes such as albumin and immunoglobulin such as peroxidase (PO
Glycoproteins such as D), erythrocytes and the like are also preferred as carriers for such purposes. In consideration of the problem of biohazard, other carriers are preferable to red blood cells.

The liposome used in the present invention may be any liposome in which a dye is encapsulated and the sugar chain compound of the present invention is immobilized on its membrane. It suffices to follow liposomes prepared in the field.

That is, for example, the preparation methods include vortex swing method, ultrasonic method, surfactant removal method, reverse phase evaporation method (REV method), ethanol injection method, ether injection method, -Pre-Vesicle
Method, French Press Extrusion (French Press
Extrusion) method, Ca ²⁺ fusion method, annealing (Annealin)
g) method, freeze-thaw fusion method, freeze-drying method, W / O / W emulsion method and the like, and SMGruner et al. [Biochemistry, 24, 28
Stable Plurilamellar Ve, reported by J. 33 (1985)]
The liposomes prepared by these methods can be used to convert a liposome produced by these methods into a membrane having a certain pore size using a French press, a pressure filter or an extruder. By passing through, liposomes having the desired properties can be obtained.

The main membrane components include natural lecithin (eg, egg yolk lecithin, soybean lecithin, etc.) and dipalmitoyl phosphatidylcholine (D) which are used as membrane components in the preparation of ordinary liposomes.
PPC), dimyristoylphosphatidylcholine (DMPC),
Distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphatidylethanolamine (DMPE), dipalmitoylphosphatidylglycerol (DPPG) , Dimyristoyl phosphatidyl glycerol (DMPG), phospholipids such as dimyristoyl phosphatidic acid (DMPA), egg yolk phosphatidyl glycerol, or glycolipids such as ganglioside glycolipid, glycosphingolipid, glyceroglycolipid, etc. One or more of them, or a combination of these with a mixed system of cholesterols and the like are all mentioned. still,
For example, it is needless to say that a liposome containing a phospholipid-containing sheep erythrocyte chloroform-extracted fraction or the like used as a constituent component of Kinsky, SC (Biochemistry, 8, 4149, 1969) can be used.

Dyes encapsulated in liposomes according to the present invention include J. liposome Res. (D. Monroe: 1 (3), 339-37).
7, 1989-90), for example, fluorescent substances such as carboxyfluorescein, fluorescein isothiocyanate, fluorescein isocyanate, tetrarhodamine isothiocyanate, 5-dimethylamino-1-naphthalenesulfonyl chloride and the like, for example, arsenazo III, 4 -
Pigments such as sodium salt of (2-pyridylazo) resorcinol, 2- (5-bromo-2-pyridylazo) -5- (N-propyl-N-sulfopropylamino) phenol, for example, luminol, isoluminol, luciferin, eosine Any liposomes such as luminescent substances such as Y, auramine O, bis (2,4,6-trichlorophenyl) oxalate, N-methylacridinium ester, etc. can be visually confirmed. Any of these may be appropriately selected in consideration of the detection method, sensitivity, and liposome stability.

The method for immobilizing the sugar chain compound of the present invention on the liposome used in the present invention may be performed according to the method for immobilizing a hapten or the like on a liposome usually used in this field. A method using a fatty acid derivative or a phospholipid derivative of a sugar chain compound or the like as a liposome membrane component (Kinsky, SC, Biochemistry, 8, 4149, 1969;
Shichijo, journal of Immunological ethods, 85, 53-63,
1985, K. Uemura, Biochemistry, vol. 11, No. 22, 4085, 197
2, T.masaki, Journal of Immunological Methods, 123, 1
9-24,1989, C. Braman, Bio.Technology, April, 349,198
4, U. Glagasigij, Chem. Pharm. Bull, 36, 1086, 1988, K. Ku
botsu, Clin. Chem, 38, 808, 1992). In addition, "Seismic Chemistry Experiment Course 5 Immunobiochemical Experiment Method,
First edition, first print, edited by: The Biochemical Society of Japan, Tokyo Kagaku Dojin Co., Ltd., pp. 144-148, published on March 14, 1986. It is also possible to prepare the liposome according to the present invention using a method of immobilizing a sugar chain compound or the like.

The crosslinking agent used in the crosslinking method includes, for example, N-succinimidyl 3- (2-pyridyldithio) propionate (SPDP), N-succinimidyl 4- (p-maleimidophenyl) butyrate (SMPB) , N-succinimidyl 4- (p-maleimidophenyl) acetate (SMPA), N-succinimidyl 4- (p-maleimidophenyl) propionate acetate (SMPP), N- (4-maleimidobutyryloxy) succinimide (GMBS) And N- (6-maleimidocaproyloxy) succinimide (EMCS).

In the sensitized particle agglomeration method, in order to support the sugar chain compound of the present invention on a carrier, an amino group which may have a substituent of the sugar chain compound of the present invention and a carrier The amino group and the functional group that reacts may be chemically bonded by a conventional method.

In the sensitized particle agglutination method, the amount of the sugar chain compound of the present invention carried on a carrier is 0.1 pg to 1 ng, preferably 0.5 to 200 pg, per unit surface area (cm ² ) of the carrier.
g, more preferably 1 to 200 pg.

In the immunochromatography method, the absorptive carrier for supporting a sugar chain compound is, among the carriers used in ordinary immunoassays, those having absorptivity and having a property of causing capillary action. For example, porous sheet-like or film-like materials, foams, woven fabrics, non-woven fabrics, knitted fabrics, and the like can be used, and these are natural, semi-synthetic or fully synthetic fibrous or other materials. And the like, for example, those obtained by molding a material having a shape in accordance with a conventional method such as paper making, film forming, foam molding, knitting, and the like. Examples of such materials include cotton, hemp, silk, cellulose, and nitrocellulose. , Cellulose acetate, rock wool, animal hair, carbon fiber, boron fiber, and other fibrous materials such as polystyrene, polypropylene, polyamide, polyvinyl alcohol, and polyvinyl acetate. Over DOO, polyester, polyacrylic acid, polyvinyl chloride, polyvinylidene chloride, synthetic polymer compounds such as polyethylene and the like.

In the immunochromatography, as a method for supporting the sugar chain compound of the present invention on an absorptive carrier, for example, a solution containing a synthetic blood group antigen on an absorptive carrier is applied, for example, by coating, dripping or spraying. Thereafter, this is dried and supported by physical adsorption, or when the absorbent carrier has a functional group that reacts with an amino group, by chemical bonding, as in the case of the sensitized particle aggregation method. A method of supporting the sugar chain compound of the present invention on an absorptive carrier and the like can be mentioned. The absorbent carrier carrying the sugar chain compound of the present invention is preferably subjected to a so-called blocking treatment in order to prevent the influence of nonspecific adsorption on the measurement. The blocking treatment may be carried out by a method generally used in this field, for example, by using an absorbent carrier carrying a synthetic antigen on a blocking agent such as albumin, globulin, casein, polyvinyl alcohol, a surfactant, etc. (For example, Tris buffer, phosphate buffer, veronal buffer, borate buffer, good buffer having a pH of about 5 to 9 and 10 to 500 mM, for example). Liquid) or the like, followed by drying.

The amount of the synthetic blood group antigen carried on the absorptive carrier varies depending on the type of blood group to be determined, the detection limit, and the like. As a supported amount per cm ² ), usually 0.01 μg to 1
mg, preferably 0.05 μg to 100 μg, more preferably 0.1 μg
g to 50 µg.

As the labeling substance, any substance can be used as long as the substance labeled with a synthetic blood group antigen can be moved by so-called capillary action, and is, for example, a radioisotope, for example, an enzyme, for example, gold colloid or iron colloid. Metal colloids such as silver and silver colloids; nonmetallic colloids such as selenium colloids; fluorescent dyes such as rhodamine B, rhodamine isothiocyanate, carboxylfluorescein, and fluorescein isothiocyanate; For example, colored latex prepared using a high-molecular polymer such as polystyrene or polyacrylamide as a raw material,
For example, colored liposomes containing fluorescent dyes and dyes as described above can be used. Among them, for example, a labeling substance capable of obtaining a visually detectable signal such as colloidal gold, selenium colloid, or colored latex is preferable, and colloidal gold is particularly preferable in terms of ease of handling and sensitivity.

The amount of the labeled synthetic blood group antigen retained on the absorbent carrier varies depending on the type of blood group to be determined, the detection limit, and the like, but the unit of the portion of the absorbent carrier on which the labeled synthetic blood group antigen is retained is described. As a retention amount per area (cm2), the amount is usually 0.01% in terms of the amount of unlabeled blood group antigen of labeled synthetic blood group antigen.
The amount is from μg to 10 mg, preferably from 0.05 μg to 1 mg, more preferably from 0.1 to 100 μg.

When the sugar chain compound of the present invention is immobilized on a carrier as described above, steric hindrance does not occur when the sugar chain compound is introduced into the carrier, and its antigenicity can be maintained after immobilization. Therefore, it is desirable that the hydrocarbon residue used as the spacer has a certain length. For example, if an alkylene group is used as the spacer, one having 2 to 10 carbon atoms, preferably 3 to 8 carbon atoms,
Among them, linear ones are preferred.

When the sugar chain compound of the present invention is supported on a carrier and used for determination of human blood type, among the compounds represented by the general formula [1], the sugar residue represented by X is 1 to 5 A monosaccharide residue is more preferable as a raw material for a human blood type determination reagent because it has a higher reactivity with an antibody than a compound in which X is a bond.

The human blood type determination reagent of the present invention contains the sugar chain compound of the present invention as a main component thereof. Usually, the sugar chain compound of the present invention is immobilized on the above-described carrier by the method described above. Take a sensitized form. In addition, preferred embodiments, use concentrations, and the like are as described above.

When the human blood type determination reagent of the present invention is a sensitizing particle agglutination reagent, latex particles, liposomes, proteins, erythrocytes, etc., to which the sugar chain compound of the present invention has been sensitized (immobilized). Is usually provided in the form of a solution suspended or dissolved in an appropriate buffer. As the buffer used for such purpose, all those usually used in this field can be used, and the buffer concentration, pH, etc. can be appropriately selected from the range usually used in this field. Good.
Further, in the buffer, commonly used in this field,
Preservatives, surfactants, for example, coagulation accelerators such as polyethylene glycol, non-specific reaction inhibitors such as magnesium ions and the like may be contained, and the concentration to be used is appropriately selected within the range usually used in this field. do it.

As described above, in the sugar chain compound of the present invention, since an amino group which may have a substituent is bonded via a divalent hydrocarbon residue, it is conventionally possible to directly bond the amino group. Not only can not be easily bonded to a carrier having a carboxyl group, an alkoxycarbonyl group, an acyloxycarbonyl group, a haloformyl group, a chlorosulfonyl group, a cyanate group, an epoxy group, a ketone group, an aldehyde group, etc. Labeling of a carboxyl group-containing sugar chain compound to a carrier, a dye, or the like, which was conventionally difficult to label, can be easily performed.

Furthermore, if a substance having a spacer of an appropriate length is used, introduction into a carrier caused by conventional methods, for example, steric hindrance is hindered, or when a liposome is used as a carrier, Problems such as inability to maintain antigen activity can be avoided.

Further, when the sugar chain compound of the present invention comprises the same antigenic site as human blood group antigen, the corresponding antibody can be produced by using the same, and the sugar chain compound of the present invention can be produced. Is used to confirm the presence of the sugar chain compound itself or its antibody, that is, by using the human blood group determination reagent produced using the sugar chain compound of the present invention, human blood group determination can be easily performed. it can.

Hereinafter, the present invention will be described in more detail with reference to Reference Examples and Examples, but the present invention is not limited thereto.

[0146]

【Example】

<Synthesis of sugar chain compound> (1) 1.04 g of 3-amino-1-propanol was dissolved in 3 ml of chloroform, and a solution of 2.0 g of methyl trifluoroacetate in chloroform (1 ml) was added. After stirring at room temperature for 2 hours, the reaction solution was purified, concentrated under reduced pressure, and CF ₃ CONH (CH ₂ ) ₃ OH
2.35 g was obtained (yield: 99.2%). Elemental analysis Calculated value for C ₅ H ₈ NO ₂ F ₃ (171.12) (%); C: 35.10, H: 4.71, N: 8.19 Actual value (%); C: 34.93, H: 4.89, N: 8.20

(2) 10 g of 6-amino-1-hexanol
Was dissolved in 5 ml of chloroform, and methyl trifluoroacetate was dissolved.
A solution of 1.1 g of chloroform (1 ml) was added. After stirring at room temperature for 5 hours, the reaction solution was purified and crystallized from diethyl ether / hexane to obtain 1.62 g of CF ₃ CONH (CH ₂ ) ₆ OH (yield: 89.0%). Mp; 39.1-40.7 ° C. Elemental Analysis _{C 8 H 14 NO 2 F 3} (213.20) Calculated (%); C: 45.07, H: 6.62, N: 6.57 Found (%); C: 44.98, H: 6.61 , N: 6.55

(3) 8-bromo-1-octanol 1.1
3 g and 1.0 g of potassium phthalimide were dissolved in DMF,
Stirred at 00 ° C. for 2 hours. The reaction solution was diluted with chloroform (250 ml), washed with saturated aqueous sodium hydrogen carbonate and saturated saline, and dried over anhydrous magnesium sulfate. After the solvent was distilled off, the residue was crystallized from hexane to give compound [1].

[0149]

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1.39 g was obtained (yield: 93.3%). Melting point: 56.7-57.4 ° C Elemental analysis Calculated value for C ₁₆ H ₂₁ NO ₃ (275.35) (%); C: 69.79, H: 7.69, N: 5.09 Actual value (%); C: 69.80, H: 7.68, N : 5.11

(4) 2- (Trimethylsilyl) ethyl synthesized from pentaacetyl-β-D-galactose according to the method described in the literature [J. Org. Chem., 53, 5629 (1988)]
2,7.8 g of 2,3,4,6-tetra-O-acetylgalactopyranoside
Was dissolved in 1.3 L of methanol, and 3.3 ml of 28% -sodium methylate was added. After stirring at room temperature overnight, cation exchange resin Amberlyst 15 was added to neutralize. After filtration, the filtrate was concentrated and the residue was treated with benzaldehyde 2
It was dissolved in 00 ml and 200 ml of formic acid. After stirring at room temperature for 1 hour, the mixture was neutralized with saturated aqueous sodium hydrogen carbonate and extracted with ethyl acetate. The organic layer was washed twice with brine and dried over anhydrous magnesium sulfate. After evaporating the solvent and purifying the residue, crystallization is carried out to give compound [2].

[0152]

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(In the formula, Ph represents a phenyl group, SE represents a 2- (trimethylsilyl) ethyl group.) 45.0 g was obtained (yield; 56.5%). Mp; from 108.5 to 109.7 ° C. Calculated elemental analysis _{C 18 H 28 O 6 Si (} 368.50) (%); C: 58.67, H: 7.66, Found (%); C: 58.64, H: 7.68,

(5) Compound [2] obtained in (4) 45.
0 g was dissolved in 207 ml of pyridine, and a solution of 48 g of benzoyl cyanide in 23 ml of acetonitrile was added thereto while cooling to 0 ° C. After stirring at 0 ° C. for 3 hours, 300 ml of methanol was added, and the mixture was further stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure,
Extracted with dichloromethane. The organic layer was washed sequentially with 0.2N hydrochloric acid, saturated aqueous sodium hydrogen carbonate and brine, and then dried over anhydrous magnesium sulfate. After evaporating the solvent, the residue is
(Eluent: hexane: ethyl acetate = 3: 1). Crystallization from hexane / ethyl acetate gave compound [3].

[0155]

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(Wherein Bz represents a benzoyl group;
And SE are the same as above. 52.0 g was obtained (yield; 90.1%). Mp; 115.3 to 117.0 ° C. Calculated elemental analysis _{C 25 H 32 O 7 Si (} 472.61) (%); C: 63.54, H: 6.82, Found (%); C: 63.54, H: 6.83,

(6) Compound [3] 52.0 obtained in (5)
g and fucose (manufactured by Aldrich) from the literature (Carbohydr.
s., 209, C1 (1991), Carbohydr. Res., 201, 31 (199
0)], methyl 2,3,4-tri-
58.7 g of O-benzyl-1-thio-β-fucosylpyranoside
It was dissolved in 2 L of a mixed solvent of 1,2-dichloromethane: DMF = 5: 1, and a mixture of 44.2 g of copper (II) bromide, 70.9 g of tetrabutylammonium bromide and 90 g of molecular sieves 4A was added thereto. After stirring at room temperature for 60 hours under a nitrogen atmosphere, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure, and the obtained residue was extracted with ethyl acetate. The organic layer was washed sequentially with saturated aqueous sodium hydrogen carbonate and brine, and dried over anhydrous magnesium sulfate. After evaporating the solvent, the residue was purified by silica gel column (eluent; hexane: ethyl acetate = 6: 1) to obtain compound [4].

[0158]

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(Wherein Bn represents a benzyl group, and Ph,
SE and Bz are the same as above. 82.4 g (yield; 84.2)
%). Calculated elemental analysis _{C 52 H 60 O 11 Si (} 889.13) (%); C: 70.25, H: 6.80, Found (%); C: 70.28, H: 6.84,

(7) Compound [4] obtained in (6)
4 g was dissolved in 5 L of a benzene: methanol = 1: 9 mixed solvent, and 1 L of 1N-sodium methylate was added. 2 at room temperature
After stirring day and night, 60 ml of acetic acid was added for neutralization, and the residue was concentrated and extracted with ethyl acetate. The organic layer was washed twice with brine and dried over anhydrous magnesium sulfate. After evaporating the solvent, the residue was purified by a silica gel column (eluent; hexane: ethyl acetate = 4: 1). After crystallization from hexane, compound [5]

[0161]

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58.8 g were obtained (yield; 80.9%). Mp; from 102.7 to 103.6 ° C. Calculated elemental analysis _{C 45 H 56 O 10 Si (} 785.02) (%); C: 68.85, H: 7.19, Found (%); C: 68.82, H: 7.20,

(8) Compound [5] obtained in (7) 1.0
g, 7.4 g of silver carbonate and 6.5 g of anhydrous calcium sulfate were dissolved in 40 ml of dichloromethane and cooled to -30 to -20 ° C. This was synthesized from 0.36 g of silver trifluoromethanesulfonate and then from pentaacetyl-β-D-galactose (manufactured by Aldrich) according to the method described in the literature [Can. J. Chem., 57, 1244 (1979)]. A solution of 1.54 g of 3,4,6-tri-O-acetyl-2-azido-2-deoxy-α-D-galactopyranosyl bromide in dichloromethane (10 ml) was added. After stirring at the same temperature for 3.5 hours, the reaction solution was filtered. The filtrate was washed twice with brine and dried over anhydrous magnesium sulfate. After evaporating the solvent,
The residue was purified by silica gel column (eluent; hexane: ethyl acetate = 3: 1) to give compound [6].

[0164]

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(In the formula, Ac represents an acetyl group, and Ph,
SE and Bn are the same as above. ) 1.37 g (yield; 97.9)
%). Calculated elemental analysis _{C 57 H 71 N 3 O 17} Si (1098.29) (%); C: 62.34, H: 6.52, N: 3.83 Found (%); C: 62.40, H: 6.59, N: 3.79

(9) 5.0 g of the compound [6] obtained by the method of (8) was dissolved in 150 ml of acetic acid, and 5 g of 10% palladium carbon was added. After stirring at room temperature for 5 days under a hydrogen atmosphere, the reaction solution was filtered. The filtrate is concentrated under reduced pressure, and the residue is subjected to a silica gel column (eluent; chloroform: methanol = 9: 1).
Was purified. Then, the obtained compound was treated with pyridine 70m
l, dissolved in 30 ml of acetic anhydride, and 10 mg of N, N-dimethylaminopyridine was added. After stirring at room temperature overnight, the reaction solution was concentrated and extracted with ethyl acetate. 0.2N-hydrochloric acid,
After washing with a saturated aqueous solution of sodium bicarbonate and brine in that order, the extract was dried over anhydrous magnesium sulfate. After evaporating the solvent, the residue was purified by a silica gel column (eluent; chloroform: methanol = 50: 1). Crystallization from diethyl ether gave compound [7].

[0167]

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3.47 g were obtained (yield; 78.9%). Melting point: 112.6-114.5 ° C Elemental analysis: calculated as C ₄₁ H ₆₃ NO ₂₃ Si (966.03) (%); C: 50.98, H: 6.57, N: 1.45, actual value (%); C: 51.10, H: 6.65 , N: 1.41,

(10) Compound [7] obtained in (9)
3.23 g was dissolved in 10 ml of dichloromethane, and 4.24 ml of boron trifluoride diethyl ether complex was added under ice cooling. After stirring under ice cooling for 3.5 hours, the reaction solution was diluted with dichloromethane. The dichloromethane layer was washed sequentially with saturated aqueous sodium hydrogen carbonate and brine, and then dried over anhydrous magnesium sulfate. After evaporating the solvent, the residue was purified by silica gel column (eluent; hexane: ethyl acetate = 1: 4) to obtain compound [8].

[0170]

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(In the formula, Ac is the same as described above.) 1.79 g was obtained (yield; 61.9%). Calculated elemental analysis _{C 36 H 51 NO 23 (865.79} ) (%); C: 49.79, H: 5.94, N: 1.62, Found (%); C: 49.84, H: 5.99, N: 1.59,

(11) 1.85 g of the compound [8] obtained by the method of (10) was dissolved in 10 ml of dichloromethane, and 3 ml of trichloroacetonitrile and 1,8-diazabicyclo [5,4,0] -7- under ice cooling. 370 μl of undecene was added. After stirring for 2 hours under ice-cooling, the reaction solution is concentrated, and the residue is subjected to a silica gel column (eluent; hexane: ethyl acetate: ethanol = 10: 5: 1).
Was purified. Powdered with hexane, compound [9]

[0173]

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(In the formula, Ac is the same as described above.) 1.55 g was obtained (yield; 71.8%). Elemental analysis Calculated value for C ₃₈ H ₅₁ N ₂ O ₂₃ Cl ₃ (1010.18) (%); C: 45.18, H: 5.09, N: 2.77, measured value (%); C: 45.11, H: 5.09, N: 2.78,

(12) 2.0 g of the compound [9] obtained by the method of (11), CF ₃ CONH (CH ₂ ) ₆ OH obtained by (2)
850 mg and 15 g of Molecular Sieves 4A were suspended in 60 ml of dichloromethane, and 1 ml of trimethylsilyltrifluoromethanesulfonic acid was added thereto under ice cooling. The reaction solution was stirred for 1 hour under ice-cooling and for 1.5 hours at room temperature, and then diluted with 100 ml of a chloroform / methanol = 5/1 mixture.
After removing the insolubles by filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by a silica gel column (eluent; hexane: ethyl acetate: ethanol = 10: 5: 1) to give compound [10].

[0176]

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(In the formula, Ac is the same as described above.) 2.03 g (yield: 96.7%) was obtained. In addition, it was confirmed by ¹ H-NMR that the target compound was obtained. Calculated elemental analysis _{C 44 H 63 N 2 O 24} F 3 (1060.98) (%); C: 49.81, H: 5.99, N: 2.64, Found (%); C: 49.87, H: 6.07, N: 2.62,

Embodiment 2 FIG. 2.0 g of the compound [10] obtained in Example 1 was
Dissolve in 160 ml, then add 0.6% 28% sodium methoxide
After stirring at room temperature overnight, add 25% aqueous ammonia
40 ml was added and the mixture was stirred at room temperature for 2 days. After the reaction solution was neutralized with acetic acid and concentrated under reduced pressure, the residue was subjected to a silica gel column (eluent; chloroform: methanol: acetic acid: water = 6: 4: 0.5: 0.
Purification was performed in 5), and the eluted fraction was concentrated under reduced pressure. The residue was purified on a gel filtration column (eluent; methanol) to give Compound [1].
1]

[0179]

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(In the formula, Ac is the same as described above.) 700 mg was obtained. (Yield: 59.3%) Note that ¹ H-NMR confirmed that the target compound was obtained. Elemental analysis Calculated value for C ₂₆ H ₄₈ N ₂ O ₁₅ (628.67) (%); C: 49.67, H: 7.70, N: 4.46, actual value (%); C: 49.69, H: 7.71, N: 4.44, FIG. 1 shows the IR spectrum of the obtained compound.

Example 3 (1) Compound [5] obtained in (7) of Example 1 2.83
g, pentaacetyl-β-D-galactose (manufactured by Aldrich) from the literature [Justus Liebigs Ann. Chem., 657, 179 (196
2)] ethyl 2,3,4,6-tetra-O-benzyl-1-thio-β-galactopyranoside synthesized according to the method described in 2.11 g
To a solution of 2 g of Molecular Sieves 4A and diethyl ether was added 2.0 ml of methyl trifluoromethanesulfonate. After stirring at room temperature for 20 hours under a nitrogen atmosphere, the reaction solution was neutralized with 5.0 ml of triethylamine. Filter off insolubles,
After the filtrate was concentrated under reduced pressure, the residue was purified by a silica gel column (eluent; hexane: ethyl acetate = 4: 1). After recrystallization from hot ethanol, compound [12]

[0182]

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(In the formula, Ph, SE and Bn are the same as described above.) 3.26 g (yield; 69.1%) was obtained. Melting point: 112.9-114.1
℃ Elemental analysis Calculated value (%) as C ₇₉ H ₉₀ O ₁₅ Si (1307.66); C: 72.56, H: 6.94, measured value (%); C: 72.55, H: 6.96,

(2) The compound [1] obtained by the method of (1)
2] Dissolve 9.54 g in 300 ml of acetic acid and add 10% -palladium carbon
9.5 g was added. After stirring at room temperature for 3 days under a hydrogen atmosphere, the reaction solution was filtered, and the filtrate was dried under reduced pressure. Next, the obtained residue was dissolved in 200 ml of pyridine and 300 ml of acetic anhydride, and 30 mg of N, N-dimethylaminopyridine was added. After stirring at room temperature overnight, the reaction solution was concentrated and extracted with chloroform. The organic layer was washed sequentially with 0.2N hydrochloric acid, saturated aqueous sodium hydrogen carbonate and brine, and then dried over anhydrous magnesium sulfate. After evaporating the solvent, the residue was purified by a silica gel column (eluent; hexane: ethyl acetate = 1: 1). Crystallization from diethyl ether gave compound [13].

[0185]

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(Wherein SE and Ac are the same as described above) 5.
62 g were obtained (yield; 70.9%). Melting point: 95.4-98.0 ° C Elemental analysis: calculated as C ₄₁ H ₆₂ O ₂₄ Si (967.01) (%); C: 50.92, H: 6.46, measured value (%); C: 50.94, H: 6.49,

(3) Compound [13] obtained in (2)
1.50 g was dissolved in 5 ml of dichloromethane, and 2.0 ml of boron trifluoride diethyl ether complex was added under ice cooling. below freezing
After stirring for 4 hours, the reaction was diluted with dichloromethane. The dichloromethane layer was washed sequentially with saturated aqueous sodium hydrogen carbonate and brine, and then dried over anhydrous magnesium sulfate. After evaporating the solvent, the residue was purified by silica gel column (eluent; hexane: ethyl acetate = 1: 2) to obtain compound [14].

[0188]

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(In the formula, Ac is the same as described above.) 1.06 g was obtained (yield; 79.1%). Elemental analysis: calculated as C ₃₆ H ₅₀ O ₂₄ (866.78) (%); C: 49.89, H: 5.81, found (%); C: 49.92, H: 5.83,

(4) The compound [1] obtained by the method of (3)
4] 1.12 g was dissolved in 5 ml of dichloromethane, and 2 ml of trichloroacetonitrile and 1,8-diazabicyclo [5,
4,0] -7-undecene (224 μl) was added. After stirring for 2 hours under ice cooling, the reaction solution is concentrated, and the residue is
(Eluent: hexane: ethyl acetate: chloroform = 20: 10:
Purified in 1). Powdered with hexane, compound [15]

[0191]

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(In the formula, Ac is the same as described above.) 1.20 g was obtained (yield; 92.0%). Elemental analysis Calculated value for C ₃₈ H ₅₀ NO ₂₄ Cl ₃ (1011.17) (%); C: 45.14, H: 4.98, N: 1.39, actual value (%); C: 45.09, H: 5.00, N: 1.41,

(5) The compound [1] obtained by the method of (4)
5] 50 mg of CF ₃ CONH (CH ₂ ) ₃ O obtained in (1) of Example 1
26 mg of H and 370 mg of Molecular Sieves 4A were suspended in 1.5 ml of dichloromethane, and 25 μl of trimethylsilyltrifluoromethanesulfonic acid was added thereto. After the reaction solution was stirred at room temperature for 1 hour, chloroform / methanol = 5.
/ 1 mixture was diluted with 5 ml. The insolubles were removed by filtration, the reaction solution was concentrated under reduced pressure, and the residue was purified by a silica gel column (eluent: hexane: ethyl acetate: ethanol = 20: 10: 1) to give compound [16].

[0194]

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Was quantitatively obtained. In addition, by ¹ H-NMR,
It was confirmed that the target compound was obtained. Elemental analysis: calculated as C ₄₁ H ₅₆ NO ₂₅ F ₃ (1019.88) (%); C: 48.29, H: 5.53, N: 1.37, found (%); C: 48.36, H: 5.61, N: 1.35,

Embodiment 4 FIG. Compound [15] obtained by method (4) of Example 3 2.0
g, 850 mg of CF ₃ CONH (CH ₂ ) ₆ OH obtained in (2) of Example 1
And molecular sieves 4A 15 g with dichloromethane 6
The suspension was suspended in 0 ml, and 1 ml of trimethylsilyltrifluoromethanesulfonic acid was added thereto. The reaction solution was stirred for 1 hour under ice-cooling and for 1.5 hours at room temperature, and then diluted with 100 ml of a chloroform / methanol = 5/1 mixture. The insoluble material was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was subjected to a silica gel column (eluent: hexane: ethyl acetate: ethanol = 10: 5:
1) Purify the compound [17]

[0197]

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1.83 g was obtained. (Yield: 87.0%) By ¹ H-NMR, it was confirmed that the target compound was obtained. Calculated elemental analysis _{C 44 H 62 NO 25 F 3} (1061.96) (%); C: 49.76, H: 5.88, N: 1.32, Found (%); C: 49.80, H: 5.91, N: 1.31,

Embodiment 5 FIG. 50 mg of the compound [15] obtained by the method of (4) of Example 3, 28 mg of the compound [1] obtained by (3) of Example 1, and molecular sieves 4A 37
0 mg was suspended in 1.5 ml of dichloromethane, and 25 μl of trimethylsilyltrifluoromethanesulfonic acid was added thereto. After the reaction solution was stirred at room temperature for 1 hour, chloroform /
The mixture was diluted with 5 ml of a mixed solution of methanol and 5/1. After filtering off the insoluble matter and concentrating the filtrate under reduced pressure, the residue was subjected to a silica gel column (eluent: hexane: ethyl acetate: ethanol = 20: 10:
Compound [18] purified by 1)

[0200]

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Was quantitatively obtained. In addition, it was confirmed by ¹ H-NMR that the target compound was obtained. Calculated elemental analysis _{C 52 H 69 NO 26 (1124.11} ) (%); C: 55.56, H: 6.19, N: 1.25, Found (%); C: 55.59, H: 6.22, N: 1.21,

Embodiment 6 FIG. Compound [1] obtained in Example 3
6] 51.0 mg was dissolved in methanol (4 ml), 28% sodium methoxide (30 μl) was added thereto, and the mixture was stirred at room temperature overnight, further, 25% aqueous ammonia (3 ml) was added, and the mixture was stirred at room temperature for 3 hours. After the reaction solution was neutralized with acetic acid and concentrated under reduced pressure, the residue was purified by a silica gel column (eluent; chloroform: methanol: acetic acid: water = 6: 4: 0.5: 0.5), and the eluted fraction was concentrated under reduced pressure. The residue is purified by a gel filtration column (eluent; methanol) to give compound [19].

[0203]

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21 mg was obtained (yield: 78.7%). In addition, it was confirmed by ¹ H-NMR that the target compound was obtained. Calculated elemental analysis _{C 21 H 39 NO 15 (545.54} ) (%); C: 46.24, H: 7.21, N: 2.57, Found (%); C: 46.25, H: 7.23, N: 2.57, obtained FIG. 2 shows the IR spectrum of the compound.

Embodiment 7 FIG. Compound [1] obtained in Example 4
7] 1.8 g was dissolved in 144 ml of methanol, and 540 μl of 28% sodium methoxide was added thereto, followed by stirring at room temperature overnight, and further adding 48 ml of 25% aqueous ammonia and stirring at room temperature for 3 days. After the reaction solution was neutralized with acetic acid and concentrated under reduced pressure, the residue was purified by a silica gel column (eluent; chloroform: methanol: acetic acid: water = 6: 4: 0.5: 0.5), and the eluted fraction was concentrated under reduced pressure. The residue is purified by a gel filtration column (eluent; methanol) to give compound [20].

[0206]

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600 mg were obtained (yield: 60.3%). In addition, it was confirmed by ¹ H-NMR that the target compound was obtained. Elemental analysis _{C 24 H 45 NO 15 (587.62} ) Calculated (%); C: 49.06, H: 7.72, N: 2.38, Found (%); C: 49.08, H: 7.72, N: 2.42, obtained The IR spectrum of the compound obtained is shown in FIG.

Embodiment 8 FIG. Compound [1] obtained in Example 5
8] 56.2 mg was dissolved in methanol (4 ml), and 28% sodium methoxide (15 μl) was added thereto. The mixture was stirred at room temperature overnight and neutralized using a cation exchange resin. After filtering off the cation exchange resin, the reaction solution was dissolved in 4 ml of methanol,
To this was added 730 mg of hydrazine acetate, and the mixture was stirred under reflux for 1 hour. The reaction solution was concentrated under reduced pressure, dimethylformamide (DMF) was added thereto to suspend, the insolubles in the reaction solution were removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified on a silica gel column (eluent; chloroform: methanol: acetic acid: water = 6: 4:
0.5: 0.5) to give compound [21]

[0209]

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20 mg was obtained (yield: 65.1%). In addition, ¹ H-N
By MR, it was confirmed that the target compound was obtained. Elemental analysis Calculated value (%) as C ₂₆ H ₄₉ NO ₁₅ (615.67); C: 50.72, H: 8.02, N: 2.28, actual value (%); C: 50.69, H: 8.09, N: 2.33, obtained The IR spectrum of the compound obtained is shown in FIG.

Embodiment 9 FIG. (1) 234 g of 6-amino-1-hexanol was dissolved in 1 L of DMF, and 296 g of phthalic anhydride and 500 ml of pyridine were added. After stirring at 120 ° C. for 1 hour, 2 L of acetic anhydride was added. After stirring at 120 ° C. for another 2 hours, the reaction solution was concentrated under reduced pressure. The residue was dissolved in ethyl acetate, washed with 0.2N hydrochloric acid, saturated aqueous sodium hydrogen carbonate and brine, and dried over anhydrous magnesium sulfate. After the solvent was distilled off, the residue was dissolved in 4 L of methanol, and 25 ml of 28% sodium methoxide was added. After stirring at room temperature for 4 hours, the mixture was neutralized using a cation exchange resin. The cation exchange resin was removed by filtration, the solvent was distilled off, and the residue was crystallized from hexane to give compound [22].

[0212]

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442 g were obtained (yield; 89.3%).

Elemental analysis: Calculated value (%) as C ₁₄ H ₁₇ NO ₃ (247.29); C: 68.00, H: 6.93, N: 5.66 Actual value (%); C: 67.98, H: 6.96, N: 5.55

(2) Compound [22] 36 obtained in (1)
4 g, 2-acetamido-2-deoxy-tetra-O-acetyl-β
382 g of -D-glucose, 117 ml of tetramethylurea and 300 g of dryalite were dissolved in 3 L of dichloromethane, and a solution of 240 g of iron (III) chloride and 400 g of dryalite in 4 L of dichloromethane was added thereto. After stirring and reacting at room temperature for 48 hours, insolubles were separated by filtration and washed with dichloromethane. The filtrate was washed with brine, saturated aqueous sodium hydrogen carbonate and brine,
It was dried over anhydrous magnesium sulfate. After distilling off the solvent,
Recrystallization from hot ethanol gives compound [23]

[0216]

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335 g were obtained (yield; 59.2%). Elemental analysis Calculated value for C ₂₈ H ₃₆ N ₂ O ₁₁ (576.60) (%); C: 58.33, H: 6.29, N: 4.86 Actual value (%); C: 58.01, H: 6.99, N: 4.87

(3) Compound [23] 36 obtained in (2)
7 g was dissolved in 6 L of methanol and 20 ml of 28% sodium methoxide was added. After stirring and reacting at room temperature for 4 hours, the precipitated crystals were collected by filtration to give compound [24].

[0219]

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260 g were obtained (yield; 90.6%). Calculated elemental analysis _{C 22 H 30 N 2 O 8} (450.49) (%); C: 58.66, H: 6.71, N: 6.22 Found (%); C: 58.65, H: 6.74, N: 6.20

(4) Compound [24] 26 obtained in (3)
0 g was suspended in 5 L of DMF, and 430 ml of benzaldehyde dimethyl acetal and 15 g of p-toluenesulfonic acid were added. After stirring and reacting at room temperature for 16 hours, the mixture was neutralized with 15 ml of 28% sodium methoxide. After the reaction solution was poured into water, the precipitated crystals were collected by filtration to give compound [25].

[0222]

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270 g were obtained (yield; 87.1%). Calculated elemental analysis _{C 29 H 34 N 2 O 8} (538.60) (%); C: 64.67, H: 6.36, N: 5.20 Found (%); C: 64.77, H: 6.33, N: 5.20

(5) Compound [25] 25 obtained in (4)
0 g was dissolved in DMF 4.5 L, and 60% -sodium hydride 28 g,
And 83 ml of benzyl bromide. After stirring and reacting at room temperature for 18 hours, methanol was added and the mixture was stirred for 30 minutes. After pouring the reaction solution into water, the precipitated crystals are collected by filtration,
Compound [26]

[0225]

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260 g were obtained (yield; 89.4%). Calculated elemental analysis _{C 36 H 40 N 2 O 8} (628.72) (%); C: 68.77, H: 6.41, N: 4.46 Found (%); C: 68.70, H: 6.46, N: 4.44

(6) Compound [26] 22 obtained in (5)
0 g, 200 g of sodium cyanoborohydride and 220 g of molecular sieves 3A were suspended in 3 L of THF, and 700 ml of a saturated hydrogen chloride diethyl ether solution was added dropwise over 30 minutes while cooling in an ice bath. After further stirring and reacting in an ice bath for 1 hour, the reaction solution was diluted with chloroform. The insolubles were removed by filtration, and the filtrate was washed with saturated aqueous sodium hydrogen carbonate and brine, and dried over anhydrous magnesium sulfate. After the solvent was distilled off, recrystallization from hot ethanol was carried out to give compound [27].

[0228]

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78 g were obtained (yield; 35.3%). Calculated elemental analysis _{C 36 H 42 N 2 O 8} (630.74) (%); C: 68.55, H: 6.71, N: 4.44 Found (%); C: 68.60, H: 6.68, N: 4.41

(7) Compound [27] 78 obtained in (6)
g, tetraacetylgalactosyl bromide 153 g, and molecular sieves 4A125 g were dissolved in dichloromethane (1.3 L), and cooled in an ice bath under cooling with silver trifluoromethanesulfonate 10 g.
700 ml of 0 g toluene solution, 25 m of 2,4,6-trimethylpyridine
l was added. The reaction solution was further reacted under stirring in an ice bath for 2 hours, and then the insoluble matter was removed by filtration. The filtrate was washed with 0.2N hydrochloric acid, saturated aqueous sodium hydrogen carbonate and brine, and dried over anhydrous magnesium sulfate. After the solvent was distilled off, the residue was dissolved in 4 L of methanol, 8 ml of 28% sodium methoxide was added, and the mixture was stirred and reacted at room temperature for 16 hours, and then neutralized with a cation exchange resin. After removing the cation exchange resin by filtration and concentrating the reaction solution under reduced pressure, the residue was purified by a silica gel column (eluent; chloroform: methanol = 9: 1) to give compound [28].

[0231]

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83.4 g was obtained (yield; 85.0%). In addition, it was confirmed by ¹ H-NMR that the target compound was obtained. Elemental analysis Calculated value for C ₄₂ H ₅₂ N ₂ O ₁₃ (792.88) (%); C: 63.62, H: 6.61, N: 3.53 Actual value (%); C: 63.55, H: 6.62, N: 3.60

Embodiment 10 FIG. 83.4 g of the compound [28] obtained in Example 9 was suspended in 830 ml of benzaldehyde, 83 g of zinc chloride was added, the mixture was stirred and reacted at room temperature for 4 hours, and the reaction solution was diluted with chloroform. The reaction solution was washed with brine, saturated aqueous sodium hydrogen carbonate and brine, and dried over anhydrous magnesium sulfate. After distilling off the solvent, crystallization from diethyl ether gave compound [29].

[0234]

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101 g was obtained quantitatively. In addition, it was confirmed by ¹ H-NMR that the target compound was obtained. Elemental analysis Calculated value for C ₄₉ H ₅₆ N ₂ O ₁₃ (880.99) (%); C: 66.80, H: 6.41, N: 3.18 Actual value (%); C: 66.89, H: 6.50, N: 3.10

Embodiment 11 FIG. 101 g of the compound [29] obtained in Example 10 is dissolved in 200 ml of pyridine, and cooled with an ice bath, and a solution of 30 g of benzoyl cyanide in 30 ml of acetonitrile is obtained.
Was added thereto, and the mixture was reacted under stirring for 4 hours under cooling, and then the reaction solution was diluted with chloroform. The reaction solution was washed with brine, 0.2N-hydrochloric acid, saturated aqueous sodium hydrogen carbonate and brine, and dried over anhydrous magnesium sulfate. After evaporating the solvent and concentrating the reaction solution under reduced pressure, the residue was purified by silica gel column (eluent: chloroform) to obtain compound [30].

[0237]

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76.7 g were obtained (yield; 74.0%). In addition, ¹ H-N
By MR, it was confirmed that the target compound was obtained. Elemental analysis _{C 56 H 60 N 2 O 14} (985.10) Calculated (%); C: 68.28, H: 6.14, N: 2.84 Found (%); C: 68.30, H: 6.15, N: 2.82

Embodiment 12 FIG. A mixture of 75 g of the compound [30] obtained in Example 11, 106 g of methyl 2,3,4-tri-O-benzyl-1-thiophose and 200 g of molecular sieves 4A was mixed with 1.5 L of a 1,2-dichloroethane / DMF = 5/1 mixed solution. Suspended in copper bromide (I
I) 76 g and 122 g of tetrabutylammonium bromide were added, and the reaction solution was stirred and reacted at room temperature for 48 hours, and then the insoluble matter was removed by filtration. The filtrate was washed with 0.2N hydrochloric acid, saturated aqueous sodium hydrogen carbonate and brine, and dried over anhydrous magnesium sulfate. After evaporating the solvent and concentrating the reaction solution under reduced pressure, the residue was purified by a silica gel column (eluent: chloroform) to obtain the compound [3]
1]

[0240]

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78.6 g were obtained (yield; 73.7%). In addition, ¹ H-N
By MR, it was confirmed that the target compound was obtained. Elemental analysis Calculated value for C ₈₃ H ₈₈ N ₂ O ₁₈ (1401.61) (%); C: 71.13, H: 6.33, N: 2.00 Actual value (%); C: 71.31, H: 6.50, N: 1.97

Embodiment 13 FIG. 67 g of the compound [31] obtained in Example 12 was dissolved in 2.6 L of methanol, 20 ml of 28% sodium methoxide was added, and the mixture was stirred and reacted at room temperature for 4 hours, and then neutralized with acetic acid. After concentrating the reaction mixture under reduced pressure, the residue was purified by a silica gel column (eluent: chloroform).
[32]

[0243]

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45 g were obtained (yield; 72.4%). In addition, ¹ H-NM
By R, it was confirmed that the target compound was obtained. Elemental analysis Calculated value for C ₇₆ H ₈₄ N ₂ O ₁₇ (1297.51) (%); C: 70.35, H: 6.53, N: 2.16 Actual value (%); C: 70.39, H: 6.50, N: 2.11

Embodiment 14 FIG. 23 g of the compound [32] obtained in Example 13, 21 g of 2-azido-2-deoxy-3,4,6-tri-O-acetylgalactosyl bromide and 70 g of molecular sieves 4A were suspended in 1 L of dichloromethane, and mercury bromide was suspended. 39g,
27 g of mercury cyanide were added. After the reaction solution was stirred and reacted at room temperature for 48 hours, insolubles were removed by filtration. The filtrate was washed with an aqueous solution of potassium iodide, a saturated aqueous solution of sodium bicarbonate and brine, and then dried over anhydrous magnesium sulfate. After the solvent was distilled off, the residue was dissolved in 2 L of methanol, and 28% -sodium methoxide 10
ml was added thereto, and the mixture was stirred and reacted at room temperature for 16 hours, and then neutralized using a cation exchange resin. The cation exchange resin was removed by filtration, and the reaction solution was concentrated under reduced pressure.

[0246]

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16 g was obtained (yield; 59.5%). In addition, ¹ H-NM
By R, it was confirmed that the target compound was obtained. Elemental analysis _{C 82 H 93 N 5 O 21} (1484.66) Calculated (%); C: 66.34, H: 6.31, N: 4.72 Found (%); C: 66.35, H: 6.37, N: 4.70

Embodiment 15 FIG. 13.7 g of the compound [33] obtained in Example 14 was dissolved in 320 ml of acetic acid, and 13 g of 10% -palladium carbon was added. The reaction vessel was purged with hydrogen gas, and the mixture was stirred and reacted at room temperature for 72 hours. After the solvent was distilled off, the residue was dissolved in pyridine (250 ml), and acetic anhydride (2) was added.
After adding 50 ml and stirring and reacting at room temperature for 16 hours, the reaction solution was concentrated under reduced pressure. The residue was washed with 0.2N hydrochloric acid, saturated aqueous sodium hydrogen carbonate and brine, dried over anhydrous magnesium sulfate, the solvent was distilled off, and the residue was purified on a silica gel column (eluent; chloroform: methanol = 100: 3). To give compound [34]

[0249]

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8 g were obtained (yield; 63.4%). In addition, ¹ H-NMR
As a result, it was confirmed that the target compound was obtained. Elemental analysis _{C 62 H 83 N 3 O 32} (1382.34) Calculated (%); C: 53.87, H: 6.05, N: 3.04 Found (%); C: 53.88, H: 6.03, N: 3.02

Embodiment 16 FIG. 7.7 g of the compound [34] obtained in Example 15 was dissolved in 500 ml of methanol, 1 ml of 28% sodium methoxide was added, and the mixture was stirred and reacted at room temperature for 24 hours. Then, the mixture was neutralized with acetic acid and concentrated under reduced pressure. . The residue was dissolved in 400 ml of methanol, 74 g of hydrazine acetate was added, and the mixture was stirred and reacted under reflux for 4 hours. The reaction solution was concentrated under reduced pressure, DMF was added to the residue for suspension, and insolubles were removed by filtration. After the filtrate was concentrated under reduced pressure, the residue was purified by a silica gel column (eluent; chloroform: methanol: acetic acid: water = 10: 10: 1: 2) to give compound [35].

[0252]

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3.2 g was obtained (yield; 63.4%). Elemental analysis
Calculated value (%) as C ₃₄ H ₆₁ N ₃ O ₂₀ (831.87); C: 49.09, H: 7.39, N: 5.05 Actual value (%); C: 49.10, H: 7.40, N: 5.10 Compound obtained The ¹ H-NMR spectrum of is shown in FIG.

Embodiment 17 FIG. 20 g of the compound [32] obtained in Example 12, 27 g of ethyl 2,3,4,6-tetra-O-benzyl-1-thiogalactose and 45 g of molecular sieves 4A
Suspended in 300 ml of a mixture of 1,2-dichloroethane / DMF = 5/1, 16 g of copper (II) bromide, 25 g of tetrabutylammonium bromide
Was added. After reacting the reaction solution with stirring at room temperature for 72 hours,
The insoluble material was removed by filtration. The filtrate was washed with 0.2N hydrochloric acid, saturated aqueous sodium bicarbonate,
After washing with brine and drying over anhydrous magnesium sulfate,
The solvent is distilled off, and the residue is purified by a silica gel column (eluent: chloroform) to give compound [36].

[0255]

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16.5 g was obtained (yield; 58.7%). In addition, ¹ H-N
By MR, it was confirmed that the target compound was obtained. Calculated elemental analysis _{C 34 H 61 N 3 O 20} (831.87) (%); C: 49.09, H: 7.39, N: 5.05 Found (%); C: 49.10, H: 7.40, N: 5.10

Embodiment 18 FIG. 18.4 g of the compound [36] obtained in Example 17 was dissolved in 700 ml of methanol, 130 g of hydrazine acetate was added, and the mixture was stirred and refluxed for 4 hours. The reaction solution was concentrated under reduced pressure, DMF was added to the residue for suspension, and insolubles were removed by filtration. The filtrate was concentrated under reduced pressure, and the residue was subjected to a silica gel column (eluent; chloroform: methanol = 9:
Compound [37] after purification in 1)

[0258]

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16.4 g was obtained (yield; 96.1%). In addition, ¹ H-N
By MR, it was confirmed that the target compound was obtained. Elemental analysis Calculated value for C ₁₀₂ H ₁₁₆ N ₂ O ₂₀ (1690.04) (%); C: 72.49, H: 6.92, N: 1.66 Actual value (%); C: 72.51, H: 6.88, N: 1.67

Embodiment 19 FIG. Compound obtained in Example 18
16 g was dissolved in 500 ml of acetic acid, and 16 g of 10% -palladium carbon was added. The reaction vessel was purged with hydrogen gas, and the mixture was stirred and reacted at room temperature for 72 hours. The solvent is distilled off, and the residue is purified by a silica gel column (eluent; chloroform: methanol: acetic acid: water = 10: 10: 1: 2) to give compound [38].

[0261]

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7 g was obtained (yield; 88.7%). Elemental analysis _{C 32 H 58 N 2 O 20} (790.81) Calculated (%); C: 48.60, H: 7.39, N: 3.54 Found (%); C: 48.61, H: 7.40, N: 3.57 to obtain The ¹ H-NMR spectrum of the compound obtained is shown in FIG.

<Judgment of Blood Group by Sensitized Particle Aggregation Method> (1) Preparation of liposome reagent sensitizing human A-type antigen sugar chain 0.18 mmol of octadecyleicosanoic acid, 0.22 mmol of N-hydroxysuccinimide and 0.22 mmol of dicyclohexylcarbodiimide are dissolved in 2.5 ml of chloroform and reacted at room temperature for 2 hours. Was. The reaction solution was diluted with 50 ml of chloroform, washed with saturated saline, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was dissolved in 3 ml of DMF, and 0.18 mmol of the compound [11] obtained in Example 2 was added, followed by stirring at room temperature for 2 hours. After concentrating the reaction solution, the residue is purified by silica gel column (eluent; chloroform: methanol = 5: 1 → 2: 1)
To obtain glycolipids. The obtained glycolipid 0.157μmo
l, dipalmitoyl phosphatidylcholine 14.2 μmol,
Dipalmitoyl phosphatidyl glycerol 1.57μmol
And 15.7 μmol of cholesterol were dissolved in 5 ml of a chloroform / dichloromethane = 1/4 mixed solution, and stirred at 200 rpm.
0mM FD & C Red No.40 (Wako Pure Chemical Industries, Ltd.) aqueous solution 1
0 ml was added dropwise, and the mixture was stirred at 250 rpm for 2 hours to form liposomes. Thereafter, the mixture was placed in boiling water for 10 minutes to completely remove the organic solvent, and then centrifuged and washed with physiological saline to obtain the same antigenicity as human type A antigen. A liposome encapsulating a red dye having an average particle size of 10 μm and having the sugar chain compound immobilized on the surface was obtained. This was appropriately diluted with physiological saline and used as a liposome reagent sensitized to human A-type antigen sugar chain.

(2) Determination of blood type To the liposome obtained in the above (1), physiological saline was added to make 4 v / v% liposome solution, and 50 μl of each was taken, and anti-A serum and anti-B serum were respectively added thereto. Each 50 μl was mixed. As a result, no aggregation was observed with the addition of anti-B serum,
With the addition of anti-A serum, liposome aggregation could be visually confirmed. FIG. 7 shows the state of the liposome solution to which anti-B serum was added, and FIG. 8 shows the state of the liposome solution to which anti-A serum was added.

Embodiment 21 FIG. (1) Preparation of anti-human erythrocyte rabbit Fab'-POD-A type sugar chain antigen complex Peroxidase (hereinafter abbreviated as POD) (manufactured by Toyobo Co., Ltd.) 30 mg was added to 1 ml of 50 mM phosphate buffer (pH 7.5). ). 5 mg of sulfosuccinimidyl 4- (p-maleimidophenyl) butyrate (Sulfo-SMPB (manufactured by Pierce Co., Ltd.)) dissolved in dimethyl sulfoxide was added thereto, and the mixture was added at 5 ° C.
Left for hours. Next, 30 mg of the compound [35] (A-type sugar chain antigen derivative) obtained in Example 16 and 10 mg of water-soluble carbodiimide (hereinafter abbreviated as WSC) were further added thereto, and the mixture was allowed to stand at room temperature for 4 hours. After immobilizing the compound on the POD, the target product was purified using a gel filtration column (Superdex200 (manufactured by Amersham Pharmacia Biotech), 50 mM PBS pH 6.0). The purified target product was reacted with an anti-human erythrocyte rabbit antibody (manufactured by ICN) which had been converted to Fab 'by a conventional method (equimolar to the added POD) overnight at 5 ° C. to react. This solution was applied to a gel filtration column (Superdex200 (manufactured by Amersham Pharmacia Biotech), 50 mM PBS p.
H6.0), 32 mg of anti-human erythrocyte rabbit Fab'-POD-A type sugar chain antigen complex (hereinafter abbreviated as Fab'-POD-A).
I got By reacting this with erythrocytes and sensitizing the erythrocytes, it can be used as a human blood type back test reagent.

(2) Preparation of antihuman erythrocyte rabbit Fab'-POD-B type sugar chain antigen complex 30 mg of POD (manufactured by Toyobo Co., Ltd.) was added to 1 ml of 50 mM phosphate buffer (p
H7.5). 5 mg of Sulfo-SMPB (manufactured by Pierce Co., Ltd.) dissolved in dimethyl sulfoxide was added thereto, and
For 2 hours. Next, 30 mg of the compound [38] (B-glycan antigen derivative) obtained in Example 19, WSC 10
mg was further added and left at room temperature for 4 hours to immobilize the compound [38] obtained in Example 19 on POD, and then the target product was purified using a gel filtration column (50 mM PBS pH 6.0). Purified target substance and anti-human erythrocyte rabbit antibody (ICN)
Was converted to Fab 'by a conventional method (equimolar to the added POD)
Was allowed to react at 5 ° C. overnight. This solution was purified with a gel filtration column (Superdex200 (manufactured by Amersham Pharmacia Biotech Co., Ltd., 50 mM PBS, pH 6.0)) and purified from anti-human erythrocyte rabbit Fab'-POD-B type sugar chain antigen complex (hereinafter, Fab ').
Abbreviated as -POD-B. ) 36 mg were obtained. This complex is useful as a reagent for determining human blood type. That is, by using this in combination with red blood cells, the human blood group can be easily determined.

(3) Determination of blood type The anti-A antibody solution containing 3% of O-type red blood cells (manufactured by Wako Pure Chemical Industries, Ltd., agglutination value 1024 times (test tube method)) was added to 50 μl and obtained in Example 27. When 7 μM of the obtained Fab′-POD-A was mixed, strong aggregation could be visually confirmed. When 7 μM of Fab′-POD-B obtained in Example 28 was mixed instead of Fab′-POD-A, no aggregation was confirmed. Conversely, an anti-B antibody solution containing 3% O-type erythrocytes (manufactured by Wako Pure Chemical Industries, Ltd.
Agglutination value 512 times (test tube method)) 50 μl, 7 μM Fab'-POD-
Aggregation was not observed when A was mixed; instead, 7 μM F
When ab'-POD-B was mixed, strong aggregation could be visually confirmed. The agglutination rate was faster than the conventional back test (slide method), and the agglutination value and score were almost the same as those of the slide method.
Table 2 shows the results. Here, W indicates that a weak agglutination reaction is observed.

[0268]

[Table 2]

A similar study was conducted using human serum containing about 3% erythrocytes instead of the anti-A (or B) antibody solution containing 3% O-type erythrocytes. , Fab'-POD-B caused aggregation, but Fab '
No aggregation occurred in the reaction with -POD-A. Also, B
In the sera of the type, aggregation occurred by reaction with Fab'-POD-A, but did not occur by reaction with Fab'-POD-B.

From the above results, it can be seen that the use of the sugar chain compound of the present invention makes it possible to easily determine a human blood type.

[0271]

Industrial Applicability The present invention provides a sugar chain compound in which an amino group which may have a substituent is bonded via a divalent hydrocarbon residue, a carrier having the sugar chain compound immobilized thereon, and human blood. The present invention provides a human antigen-sensitizing particle, a reagent for determining a human blood group comprising the sugar chain compound, and a method for determining a human blood group using the sugar chain compound. It can be easily bonded to a carrier having a carboxyl group, an alkoxycarbonyl group, an acyloxycarbonyl group, a haloformyl group, a chlorosulfonyl group, a cyanate group, an epoxy group, a ketone group, an aldehyde group, etc., which could not be bonded. Therefore, when the sugar chain compound comprises the same antigenic site as the human blood group antigen, it can be used as an immunogen for producing the corresponding antibody, and the present invention Using a sugar chain compound to confirm the presence of the sugar chain compound itself or its antibody, that is, by using the sugar chain compound of the present invention as a human blood group determination reagent, the determination of human blood group It can be easily performed without causing a problem or the like.

[0272]

[Brief description of the drawings]

FIG. 1 is an IR spectrum of compound [11] obtained in Example 2.

FIG. 2 is an IR spectrum of compound [19] obtained in Example 6.

FIG. 3 is an IR spectrum of compound [20] obtained in Example 7.

FIG. 4 is an IR spectrum of compound [21] obtained in Example 8.

FIG. 5 shows ¹ H—N of compound [35] obtained in Example 16.
MR spectrum.

FIG. 6 shows ¹ H—N of compound [38] obtained in Example 19.
MR spectrum.

FIG. 7 is a micrograph of a liposome solution to which anti-B serum obtained in Example 21 has been added.

FIG. 8 is a micrograph of a liposome solution to which anti-A serum obtained in Example 21 has been added.

 ──────────────────────────────────────────────────続 き Continued on the front page F term (reference) 2G045 AA09 CA25 DA30 FB03 4C057 BB03 BB04 CC03 DD03 JJ09 4C085 AA08 BB21 CC33 DD51 GG10

Claims (12)

[Claims] A sugar chain compound in which an amino group which may have a substituent is bonded via a divalent hydrocarbon residue. 2. The sugar chain compound according to claim 1, wherein the divalent hydrocarbon residue is an alkylene group. 3. The sugar chain compound according to claim 1, comprising a sugar chain consisting of 2 to 8 monosaccharide residues which may have a substituent. 4. The sugar chain compound according to claim 1, wherein the sugar chain comprises the same antigen site as the human blood group antigen. 5. The method according to claim 1, wherein the human blood group antigen is human A type antigen or human B type.
The compound according to claim 4, which is a type antigen. 6. A sugar chain compound represented by the general formula [1]: (Wherein, R ¹ represents a hydroxyl group or -NHCOCH _3, R ²
Represents an amino group which may have a substituent, X represents a bond or a sugar residue, and E represents a divalent hydrocarbon residue. The compound according to claim 1, which is represented by the formula: 7. The sugar chain compound according to claim 6, wherein the divalent hydrocarbon residue represented by E is an alkylene group. 8. The sugar chain compound according to claim 6, wherein the sugar residue represented by X consists of 1 to 5 monosaccharide residues. 9. A carrier on which the sugar chain compound according to claim 1 is immobilized. 10. A human blood group antigen-sensitized particle having the sugar chain compound according to claim 4 immobilized thereon. 11. A reagent for determining a human blood group, comprising the sugar chain compound according to any one of claims 4 to 8. 12. A method for determining a human blood group, comprising using the sugar chain compound according to any one of claims 4 to 8 as an antigen.