JP5327945B2 - Glycoside and method for producing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for easily producing a glucoside from an unprotected sugar. <P>SOLUTION: The production method includes the use of a haloformamidinium derivative as a dehydrating condensation agent and a sugar having a free hemiacetal hydroxy group as a starting material and the production of the corresponding glucoside in an aqueous solution in one step. The method can be applied to a longer sugar chain and is useful for the production of substances of various uses such as a physiologically active oligosaccharide, a carrier for a drug delivery system, a surfactant, a sugar chain medicine, a reagent for determining enzyme activity, a skin lightening agent, a sugar chain synthesis reagent with an enzyme catalyst, a glycopeptide, a glycoprotein, a sugar chain polymer and a sugar chain chip, and for improving the water-solubility of a fat-soluble molecule, etc. <P>COPYRIGHT: (C)2010,JPO&amp;INPIT

Description

  The present invention relates to a method for producing a glycoside from an unprotected sugar in an aqueous solution.

A glycoside obtained by adding a sugar chain to a compound improves the water solubility of a fat-soluble molecule [Japanese Patent Laid-Open No. 2003-310294
Gazette (patent document 1)], enzyme activity measuring reagent [JP 9-275995 gazette (patent document 2)], whitening agent [Cesk. Farm., 34, 174, 1985 (non-patent document 1), JP-A-60-16906 (Patent Document 3), JP-A-60-56912 (Patent Document 4) and the like), enzyme-catalyzed sugar chain synthesis reagents [T. Murata, T. Usui: Enzymatic synthesis of important oligosaccharide units involved in N- and O-linked glycans, Trends in Glycosci.Glycotechnol., 12, 161-174,
2000 (Non-patent Document 2)], sugar chain chips [Japanese Patent Laid-Open No. 2003-083969 (Patent Document 5)], etc. have a wide range of industrial uses, and simplification of the manufacturing process is an essential technical problem. In the conventional technique, a method using organic chemical synthesis, a method utilizing an enzymatic reaction, or a method using a combination thereof is applied in accordance with the structure of a desired compound.

As a basic structure of a glycoside, it is general to take a structure in which a reducing end of a sugar and a non-sugar moiety are dehydrated and condensed. Simply, if a dehydrating condensing agent is used, a bond is formed between the reducing end of the sugar and the non-sugar moiety. In the conventional techniques, a product in which p-nitrophenol is introduced can be obtained by using a Mitsunobu reagent for a monosaccharide or disaccharide in a non-aqueous organic solvent.

However, in this technique, most of the sugars more than trisaccharide are not dissolved in an organic solvent, and therefore, this technique is substantially impossible to synthesize oligosaccharide glycosides. Therefore, an innovative method for producing glycosides using water as a main solvent has been desired.
As a technique that can achieve dehydration condensation in an aqueous solution, a method for synthesizing a sugar oxazoline derivative [Japanese Patent Application No. 2007-059478 (Patent Document 6)]

が報告されている。 And anhydro sugar synthesis method (Japanese Patent Application No. 2007-254281 (Patent Document 7))

Has been reported.

JP 2003-310294 A Japanese Patent Laid-Open No. 9-275995 Japanese Unexamined Patent Publication No. 60-16906 JP 60-56912 A Japanese Patent Laid-Open No. 2003-083969 Japanese Patent Application No. 2007-059478 Japanese Patent Application No. 2007-254281 Cesk. Farm., 34, 174, 1985 T. Murata, T. Usui: Enzymatic synthesis of important oligosaccharide units involved in N- and O-linked glycans, Trends in Glycosci. Glycotechnol., 12, 161-174, 2000

  In glycoside synthesis, development of a technique for synthesizing glycosides from unprotected sugars simply and gently without using steps such as protection and deprotection is required. In order to greatly improve the sugar chain production technology, provision of means for obtaining a desired glycoside by a one-step reaction in a solvent mainly composed of water, which is a good solvent for sugar, is required. In particular, development of a technique for synthesizing useful glycosides such as azide sugar and fluorinated sugar from an unprotected sugar in an aqueous solution is required.

As a result of diligent research on the glycoside synthesis method, the present inventors have succeeded in finding that haloformamidinium derivatives can be directly synthesized using unprotected sugar as a starting material by using a haloformamidinium derivative as a dehydrating condensing agent. The present invention has been completed. In the present invention, a sugar chain production technique is greatly improved, and a means for obtaining a desired glycoside by a one-step reaction in a solvent mainly composed of water, which is a good solvent for sugar, is provided.
In the present invention, the following modes are provided.

（上式中、R¹、R²、R³及びR⁴は、それぞれ同一でも異なっていてもよく、互いに独立に、水素原子、ヒドロキシ基、ヒドロキシメチル基、カルボキシメチル基、カルボアルコキシメチル基、アミノ基、アセチルアミノ基を含めたアシルアミノ基、カルボキシ基、アミド
基、PO₄ ^2-、SO₄ ^-、アルキル基、アシル基、アルコキシ基、アシロキシ基、グルコース、
ガラクトース、マンノース、N-アセチルグルコサミンを含めた単糖のの糖残基、セロオリゴ糖、ラミナリオリゴ糖、キトオリゴ糖、キシロオリゴ糖を含めたオリゴ糖のの糖残基及び多糖のの糖残基からなる群から選択されたものである）
で表される無保護糖と一般式MZ（式中、Zは、水素原子、ハロゲン原子、アジド基を含め
た窒素原子含有基、炭化水素基を含めた炭素原子含有基、炭化水素基で置換されていてよいオキシ基を含めた酸素原子含有基、炭化水素基で置換されていてよいリン原子含有基、及び炭化水素基で置換されていてよいチオ基を含めた硫黄原子含有基からなる群から選択されたもので、Mは金属、-OH、アンモニウム又は水素原子である）の化合物とを、一般式(2)：

（上式中、R⁵、R⁶、R⁷及びR⁸は、それぞれ同一でも異なっていてもよく、互いに独立に、置換されていてもよいアルキル基、置換されていてもよいアルケニル基及び置換されていてもよいアリール基からなる群から選択されたもので、R⁵とR⁷あるいはR⁶とR⁸で環を形成していてよいし、あるいはR⁵とR⁶あるいはR⁷とR⁸で環を形成していてよく、Xはハロゲン
原子であり、そしてY^-は陰イオンである）
で表される脱水縮合剤の存在下で処理して、一般式(3)：

（上式中、Z、R¹、R²、R³及びR⁴は、上記と同様の意味を有するものである）
で表される配糖体を得ることを特徴とする配糖体の製造方法。
〔２〕 Yが、ハロゲン原子、OH、BF₄、またはPF₆であり、一般式(2)の脱水縮合剤がハロホルムアミジニウム誘導体であり、その一般式(2)の脱水縮合剤と一般式(1)の糖との反応を、水を含む溶媒中で行うことを特徴とする上記〔１〕に記載の配糖体の製造方法。
〔３〕 一般式(2)で表される脱水縮合剤が、2-クロロ-1,3-ジメチルイミダゾリニウムクロリド、2-クロロ−1,3−ジメチルイミダゾリニウムヘキサフルオロホスファート、1-(クロロ-1-ピロリジニルメチレン)ピロリジニウムヘキサフルオロホスファート、1-(クロ
ロ-1-ピロリジニルメチレン)ピロリジニウムテトラフルオロボラート、クロロ-N,N,N',N'-テトラメチルホルムアミジニウムヘキサフルオロホスファート、N,N,N',N'-テトラメチ
ル-O-(N-スクシンイミジル)ウロニウムテトラフルオロボラート、O-(ベンゾトリアゾール-1-イル)-N,N,N',N'-テトラメチルウロニウムテトラフルオロボラート、O-(ベンゾトリアゾール-1-イル)-N,N,N',N'-テトラメチルウロニウムヘキサフルオロホスファート、O-(7-アザベンゾトリアゾール-1-イル)- N,N,N',N'-テトラメチルウロニウムテトラフルオロボラート、(4R,5R)-2-クロロ-1,3-ジメチル-4,5-ジフェニル-1-イミダゾリニウムクロリド
、及び(4S,5S)-2-クロロ-1,3-ジメチル-4,5-ジフェニル-1-イミダゾリニウムクロリドか
らなる群から選択されたものであることを特徴とする上記〔１〕又は〔２〕に記載の配糖体の製造方法。
〔４〕 一般式(3)（式中、Zは、アジド基、ハロゲン原子、及びアリールチオ基からなる群から選択されたもので、R¹、R²、R³及びR⁴は、上記と同様の意味を有するものである
）で表される配糖体を得ることを特徴とする上記〔１〕〜〔３〕のいずれか一に記載の配糖体の製造方法。 [1] General formula (1):

(In the above formula, R ¹ , R ² , R ³ and R ⁴ may be the same or different, and independently of each other, a hydrogen atom, a hydroxy group, a hydroxymethyl group, a carboxymethyl group, a carboalkoxymethyl group, Amino group, acylamino group including acetylamino group, carboxy group, amide group, PO ₄ ²⁻ , SO ₄ ⁻ , alkyl group, acyl group, alkoxy group, acyloxy group, glucose,
A group consisting of sugar residues of monosaccharides including galactose, mannose and N-acetylglucosamine, sugar residues of oligosaccharides including cellooligosaccharides, laminary oligosaccharides, chitooligosaccharides, xylooligosaccharides, and sugar residues of polysaccharides Selected from)
And a general formula MZ (wherein Z is substituted with a hydrogen atom, a halogen atom, a nitrogen atom-containing group including an azide group, a carbon atom-containing group including a hydrocarbon group, or a hydrocarbon group) A group consisting of an oxygen atom-containing group including an optionally substituted oxy group, a phosphorus atom-containing group optionally substituted with a hydrocarbon group, and a sulfur atom-containing group including a thio group optionally substituted with a hydrocarbon group And M is a metal, —OH, ammonium or a hydrogen atom), and a compound of the general formula (2):

(In the above formula, R ⁵ , R ⁶ , R ⁷ and R ⁸ may be the same or different, and independently of each other, an optionally substituted alkyl group, an optionally substituted alkenyl group and a substituted group. R ⁵ and R ⁷ or R ⁶ and R ⁸ may form a ring, or R ⁵ and R ⁶ or R ⁷ and R ^8. And X may be a halogen atom and Y ⁻ is an anion)
In the presence of a dehydration condensation agent represented by general formula (3):

(In the above formula, Z, R ¹ , R ² , R ³ and R ⁴ have the same meaning as above)
A method for producing a glycoside, comprising obtaining a glycoside represented by the formula:
[2] Y is a halogen atom, OH, BF ₄ , or PF ₆ , the dehydration condensing agent of the general formula (2) is a haloformamidinium derivative, and the dehydration condensing agent of the general formula (2) The method for producing a glycoside according to the above [1], wherein the reaction with the saccharide of formula (1) is carried out in a solvent containing water.
[3] The dehydrating condensing agent represented by the general formula (2) is 2-chloro-1,3-dimethylimidazolinium chloride, 2-chloro-1,3-dimethylimidazolinium hexafluorophosphate, 1- (Chloro-1-pyrrolidinylmethylene) pyrrolidinium hexafluorophosphate, 1- (chloro-1-pyrrolidinylmethylene) pyrrolidinium tetrafluoroborate, chloro-N, N, N ', N'- Tetramethylformamidinium hexafluorophosphate, N, N, N ', N'-tetramethyl-O- (N-succinimidyl) uronium tetrafluoroborate, O- (benzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium tetrafluoroborate, O- (benzotriazol-1-yl) -N, N, N', N'-tetramethyluronium hexafluorophosphate, O- ( 7-azabenzotriazol-1-yl)-N, N, N ', N'-tetramethyluronium tetra Fluoroborate, (4R, 5R) -2-chloro-1,3-dimethyl-4,5-diphenyl-1-imidazolinium chloride, and (4S, 5S) -2-chloro-1,3-dimethyl- The method for producing a glycoside according to [1] or [2] above, which is selected from the group consisting of 4,5-diphenyl-1-imidazolinium chloride.
[4] General formula (3) (wherein Z is selected from the group consisting of an azide group, a halogen atom and an arylthio group, and R ¹ , R ² , R ³ and R ⁴ are the same as above) The method for producing a glycoside according to any one of [1] to [3] above, wherein the glycoside represented by (1) is obtained.

According to the production method of the present invention, the degree of polymerization is 100 from monosaccharide without using an extra organic solvent.
Those glycosides can be easily produced up to the above polysaccharides. In the present invention, a glycoside, for example, a sugar derivative such as azide sugar or fluorinated sugar can be synthesized from an unprotected sugar by a simple and mild technique and in a single step and in a good yield. It can be applied to longer sugar chains, and it becomes a technology that can convert various and various sugars (oligosaccharides and polysaccharides, and sugars including branched sugar chains) into various sugar compounds and glycosylated compounds. , Bioactive oligosaccharides, drug delivery system carriers, surfactants, sugar chain drugs, enzyme activity measurement reagents, whitening agents, enzyme-catalyzed sugar chain synthesis reagents, glycopeptides, glycoproteins, sugar chains, sugar chains It is useful for manufacturing substances for various uses such as chips, and improving water solubility of fat-soluble molecules.
Other objects, features, excellence and aspects of the present invention will be apparent to those skilled in the art from the following description. However, it is understood that the description of the present specification, including the following description and the description of specific examples and the like, show preferred embodiments of the present invention and are presented only for explanation. I want. Various changes and / or modifications (or modifications) within the spirit and scope of the present invention disclosed herein will occur to those skilled in the art based on the following description and knowledge from other parts of the present specification. Will be readily apparent. All patent documents and references cited herein are cited for illustrative purposes and are not to be construed as a part of this specification. is there.

The present invention provides a method for synthesizing a glycoside or a glycosylated compound using an unprotected sugar as a starting material. In this invention, the use of the novel compound obtained by that and also this glycoside is provided.
The glycoside is not particularly limited as long as it is derived from a sugar, for example, has an activity that functions as a sugar chain-containing reagent, but is preferably a hemi such as an unprotected sugar or an unprotected sugar chain. It is synthesized from a sugar having an acetal hydroxy group and includes, for example, a glycoside represented by the above general formula (3).
The glycoside represented by the general formula (3) is produced by treating a sugar having a hemiacetal hydroxy group represented by the general formula (1) with a haloformamidinium derivative represented by the general formula (2) that is a dehydrating condensing agent. it can.

In the present specification, the “alkyl group” may be any of linear or branched, and cyclic, for example, C _1-22 alkyl (for example, methyl, ethyl, propyl, isopropyl, butyl, Isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, tert-pentyl, hexyl, heptyl, octyl, nonyl, decanyl, hexadecanyl, eicosanyl, etc.), cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc. Cyclic alkyl groups, and the like, preferably C _1-6 alkyl (for example, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, pentyl, etc.), more preferably C _1-4 alkyl. (For example, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, etc.) I can get lost.
In the present specification, the “acyl” of the “acylamino group” may include a group derived from an oxo acid such as a carboxylic acid, and examples thereof include an optionally substituted hydrocarbon group —CO— group. A carboxylic acid acyl group, and more specifically, an optionally substituted C _1-10 acyl group. Here, the optionally substituted hydrocarbon group includes, for example, an _optionally substituted C _1-22 alkyl group, an _optionally substituted C _2-10 alkenyl group, and an _optionally substituted C _{2− 10 represents an} alkynyl group, an optionally substituted aryl group, an _optionally substituted C _7-12 aralkyl group, an _optionally substituted heterocyclic group and the like. As the C _1-22 alkyl group, those described above can be used. Examples of the C _2-10 alkenyl group include vinyl, allyl, 2-butenyl, and 1-pentenyl groups. Examples of the C _2-10 alkynyl group include 1-ethynyl, propargyl, 2-butynyl, 1-pentynyl group and the like. Examples of the C _1-10 acyl group include a linear C _1-10 acyl such as formyl, acetyl, propionyl, etc., as well as a methylene group at the terminal end of the above-mentioned alkyl group as a C _1-10 alkyl group. An acyl group obtained by substituting the group is used. As the aryl group, 1-naphthyl, 2-naphthyl, 2-biphenylyl, 3-biphenylyl, 4-biphenylyl, 2-anthryl, 3-indenyl, 5-fluorenyl and the like are used. As the C _7-12 aralkyl group, benzyl, 1-phenethyl, 2-phenethyl, 1-naphthylmethyl, 2-naphthylmethyl and the like are used. The heterocyclic group may have 1 to 4 heteroatoms (selected from oxygen atom, nitrogen atom, sulfur atom, etc.) which may be the same or different, saturated or unsaturated. Monocyclic or condensed ring may be included, and for example, 2-pyridyl, 3-pyridyl, 4-pyridyl and the like are used. In the present specification, as the “acyl group”, those described above for “acyl” can be used. In the present specification, as the “acyl group” part of the “acyloxy group”, those described above for “acyl” can be used.

In the present specification, examples of the “halogen atom” include fluorine, chlorine, bromine, iodine and the like, preferably fluorine. In the present specification, the “nitrogen atom-containing group” includes an azide group, an azo group, a tetraazonium group, a nitro group, other known nitrogen atom-containing groups in the art, and the nitrogen mentioned in the present specification. Any group containing an atom may be considered to be included. In the present specification, examples of the “carbon atom-containing group” include an optionally substituted hydrocarbon group and the like, as well as those known in the art as carbon atom-containing groups, and the carbon atoms referred to in the present specification. Any group containing can be considered to be included. In the present specification, the “phosphorus atom-containing group” includes residues of phosphoric acid, phosphonic acid, phosphinic acid, phosphorous acid, phosphonous acid, phosphinic acid, residues of derivatives thereof, and the like. In the phosphorus atom-containing group, an optionally substituted hydrocarbon group may be directly bonded to the phosphorus atom, or may be bonded to the phosphorus atom via an oxygen atom, a sulfur atom, and / or a nitrogen atom. It may be bonded. The phosphorus atom-containing group may be considered to include all groups known in the art as phosphorus atom-containing groups and groups containing a phosphorus atom referred to in the present specification. In the present specification, examples of the “sulfur atom-containing group” include a thio group which may be substituted with a hydrocarbon group, as well as what is known in the art as a sulfur atom-containing group, and is referred to in the present specification. Any group containing a sulfur atom may be considered to be included. Examples of the sulfur atom-containing group include an optionally substituted arylthio group (eg, nitrophenylthio group).
In connection with the compound of general formula MZ, the “metal” in M is known in the art as a metal, for example, an alkali metal (eg, lithium, sodium, potassium, cesium, etc.), an alkaline earth metal (Eg, magnesium, calcium, barium, etc.), transition metals, aluminum, zinc, lead, tin, mercury and the like. Examples of the compound represented by the general formula MZ include aromatic thiol compounds such as alkali metal azide, alkali metal fluoride and thiophenol, and diketone compounds such as 2,4-pentanedione.

In the present specification, in relation to general formula (2) and the like, “an alkyl group which may be substituted” in “
Examples of the “alkyl group” include those similar to the above. The “alkenyl group” in the “optionally substituted alkenyl group” may be either linear or branched, for example, C _2-24 alkenyl (eg, vinyl, allyl, isopropenyl, 1- Butenyl, 2-butenyl, 3-butenyl, 2-methyl-2-propenyl, 1-methyl-2-propenyl, 2-methyl-1-propenyl and the like. Examples of the “aryl group” in the “optionally substituted aryl group” include C _6-14 aryl (for example, phenyl, 1-naphthyl, 2-naphthyl, 2-biphenylyl, 3-biphenylyl, 4-biphenylyl, 2-anthryl, 3-indenyl, 5-fluorenyl, etc.) and the like, preferably a phenyl group.

In addition, the above “optionally substituted hydrocarbon group”, “optionally substituted alkyl group”, “optionally substituted alkenyl group”, “optionally substituted aryl group” and “substituted” "Hydrocarbon group", "alkyl group", "alkenyl group" in "optionally substituted heterocyclic group" (including other cases where the modifier "optionally substituted" is attached), " “Aryl group” and “heterocyclic group” (including other corresponding groups) are optionally substituted by one or more (eg, 1-5) substituents which may be the same or different. The “substituent” in the case of the substitution may be a substituent known in the art, and may be selected from, for example, the following:

Oxo, thioxo, imino optionally having substituent, halogen atom (eg, fluorine, chlorine, bromine, iodine, etc.), C _1-3 alkylenedioxy (eg, methylenedioxy, ethylenedioxy, etc.), Nitro, cyano, C _1-6 alkyl, C _2-6 alkenyl, carboxy C _2-6 alkenyl (eg, 2-carboxyethenyl, 2-carboxy-2-methylethenyl, etc.), C _2-6 alkynyl, C _{3- 6} cycloalkyl, C _6-14 aryl (eg, phenyl, 1-naphthyl, 4-biphenylyl, 2-anthryl, etc.), C _1-8 alkoxy, C _1-6 alkoxy-carbonyl-C _1-6 alkoxy (eg, Ethoxycarbonylmethyloxy etc.), hydroxy, C _6-14 aryloxy (eg, phenyloxy etc.), C _7-16 aralkyloxy (eg benzyloxy etc.), mercapto, C _1-6 alkylthio, C _6-14 arylthio (Eg, phenylthi E), C _7-16 aralkylthio (eg, benzylthio, etc.), amino, mono-C _1-6 alkylamino (eg, methylamino, ethylamino, etc.), mono-C _6-14 arylamino (eg, phenyl) Amino), di-C _1-6 alkylamino (eg, dimethylamino, diethylamino, etc.), di-C _6-14 arylamino (eg, diphenylamino, etc.),

Formyl, carboxy, C _1-6 alkyl-carbonyl (eg, acetyl, propionyl, etc.), C _3-6 cycloalkyl-carbonyl (eg, cyclopropylcarbonyl, etc.), C _1-6 alkoxy-carbonyl (eg, methoxycarbonyl, Ethoxycarbonyl, propoxycarbonyl, tert-butoxycarbonyl, etc.), C _6-14 aryl-carbonyl (eg, benzoyl, etc.), C _7-16 aralkyl-carbonyl (eg, phenylacetyl, etc.), C _6-14 aryloxy-carbonyl (Eg, phenoxycarbonyl, etc.), C _7-16 aralkyloxy-carbonyl (eg, benzyloxycarbonyl, etc.), 5- or 6-membered heterocyclic carbonyl (eg, nicotinoyl, thenoyl, furoyl, morpholinocarbonyl, thiomorpholinocarbonyl, piperazine- 1-ylcarbonyl, pyrrolidin-1-ylcarbonyl, etc.), Carbamoyl, thiocarbamoyl, mono-C _1-6 alkyl-carbamoyl (eg, methylcarbamoyl), di-C _1-6 alkyl-carbamoyl (eg, dimethylcarbamoyl), C _6-14 aryl-carbamoyl (eg, phenyl) Carbamoyl, etc.), 5- or 6-membered heterocyclic carbamoyl (eg, 3-pyridylcarbamoyl, 2-thienylcarbamoyl, etc.),

C _1-6 alkylsulfonyl (eg, methylsulfonyl), C _6-14 arylsulfonyl (eg,
Phenylsulfonyl, etc.), C _1-6 alkylsulfinyl (eg, methylsulfinyl, etc.), C _6-14 arylsulfinyl (eg, phenylsulfinyl, etc.), formylamino, C _1-6 alkyl-carbonylamino (eg, acetylamino, etc.) ), C _6-14 aryl-carbonylamino (eg, benzoylamino, etc.), C _1-6 alkoxy-carbonylamino (eg, methoxycarbonylamino, etc.), C _1-6 alkylsulfonylamino (eg, methylsulfonylamino, etc.) C _6-14 arylsulfonylamino (eg, phenylsulfonylamino, etc.), C _1-6 alkyl-carbonyloxy (eg, acetoxy, etc.), C _6-14 aryl-carbonyloxy (eg, benzoyloxy, etc.), C _{1 -6} alkoxy - carbonyloxy (e.g., methoxycarbonyloxy, etc.), mono--C _1-6 alkyl - carbamoyloxy Examples, methylcarbamoyloxy, etc.), di -C _1-6 alkyl - carbamoyloxy (e.g., dimethylcarbamoyloxy, etc.), C _6-14 aryl - carbamoyloxy (e.g., phenylcarbamoyloxy, etc.), nicotinoyloxy oxy substituent 5- to 7-membered saturated cyclic amino, which may have a 5- to 10-membered aromatic heterocyclic group (eg, 2-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-quinolyl, 4-quinolyl) , 8-quinolyl, 4-isoquinolyl, 1-indolyl, 3-indolyl, 2-benzothiazolyl, 2-benzo [b]
Thienyl, 2-benzo [b] furanyl, 3-benzo [b] furanyl, etc.), sulfo, sulfamoyl, sulfinamoyl, sulfenamoyl and the like. In the substituents listed here, the “alkyl part” (including the alkyl part in alkoxy), “alkylene part”, “alkenyl part”, “alkynyl part”, “aryl part”, and “heterocyclic part” Optionally, it may be substituted with one or more substituents, in which case the substituents may be groups as described above. In the description of the above “substituent”, the substituent in the case of “may have a substituent” is also a group as described above.

In the case of “R ⁵ and R ⁷ or R ⁶ and R ⁸ form a ring”, the “ring” is a nitrogen atom to which R ⁵ or R ⁷ is bonded, or R ⁶ or R ⁸ is bonded. A 5-7 membered ring formed by a carbon chain which together with the nitrogen atom may further contain one or more oxygen, nitrogen and / or sulfur atoms, for example, It may be an imidazoline ring, a benzimidazoline ring, a hydropyrimidine ring, or the like. The “ring” in the case of “R ⁵ and R ⁶ or R ⁷ and R ⁸ form a ring” is the nitrogen atom to which R ⁵ and R ⁶ are bonded, or R ⁷ and R ⁸ are bonded. A 5-7 membered ring formed by a carbon chain which together with the nitrogen atom may further contain one or more oxygen, nitrogen and / or sulfur atoms, for example, It may be a pyrrolidine ring, a piperidine ring, a piperazine ring, a morpholine ring, a thiomorpholine ring, or the like. X
Is a halogen atom, for example, a chlorine atom, a bromine atom or an iodine atom.
Y ⁻ is not particularly limited as long as it is an anion, but suitable Y is, for example,
Halogen atoms such as chlorine atom, bromine atom and iodine atom, OH, BF ₄ , PF _{6 and the} like can be mentioned.

  In the present specification, the sugar residue may refer to those derived from sugar. In the present specification, the term “sugar” may be understood to include saccharides, saccharides, complex carbohydrates, etc., and “saccharide” refers to a monosaccharide, a small saccharide condensed with a plurality of monosaccharides (disaccharide, (Including oligosaccharides) and polysaccharides. Sugars are polyhydroxy aldehydes, polyhydroxy ketones, and derivatives thereof having approximately the same number of oxygen atoms as carbon atoms (for example, amino sugars having amino groups, aldehyde groups, or primary hydroxy groups become carboxyl groups). Carboxylic acid, polyhydric alcohol in which an aldehyde group or a ketone group is a hydroxy group, and the like, and their condensation polymers. “Carbohydrate” may be understood to refer to a substance having a saccharide as a main component, and includes simple sugars and other substances (including proteins, lipids, synthetic polymers, etc.) consisting only of sugar. You can think of things as complex carbohydrates.

The sugar of the present invention is not particularly limited by the origin and origin of the substance, but can be obtained from nature, genetically engineered by animal cells, plant cells, microorganisms, etc., or enzymatically produced Those produced by fermentation, or those artificially chemically synthesized may be included. Sugars may include monosaccharides, disaccharides, oligosaccharides, polysaccharides, and monosaccharides include glucose, galactose, mannose, glucosamine, N-acetylglucosamine, galactosamine, N-acetylgalactosamine, mannosamine, N-acetylmannos Examples of the disaccharide include maltose, isomaltose, lactose, lactosamine, N-acetyllactosamine, cellobiose, melibiose, N, N'-diacetylchitobiose, hyaluronic acid disaccharide And glycosaminoglycan disaccharide.

Oligosaccharides include oligosaccharides in the usual sense composed of 2 or more monosaccharides, and usually composed of 2 to 30 monosaccharides, typically 2 to 20 Examples include monosaccharides, homo-oligomers composed of glucose, galactose, mannose, glucosamine, N-acetylglucosamine, fructose, etc., or glucose, galactose, mannose, glucosamine, N-acetylglucosamine, fructose, Examples include hetero-oligomers composed of two or more components such as sialic acid, such as maltooligosaccharide, isomaltooligosaccharide, lactoligosaccharide, lactosamine oligosaccharide, N-acetyllactosamine oligosaccharide, cellooligosaccharide, melivio oligosaccharide, N- Acetylchitotriose, N-acetylchitotetraose, N-acetyl Examples include chitopentaose. Also, glycosaminoglycan oligosaccharides such as hyaluronic acid oligosaccharides (for example, hyaluronic acid disaccharide and hyaluronic acid tetrasaccharide described above), chondroitin sulfate oligosaccharides (for example, chondroitin sulfate A oligosaccharide, chondroitin sulfate C oligosaccharide) And keratan sulfate oligosaccharides, heparin oligosaccharides, heparan sulfate oligosaccharides, and the like.

Examples of polysaccharides include those found in a wide range of organisms such as animals, plants (including seaweed), insects, and microorganisms. For example, sialo-complex sugar chains, N-linked sugar chains, O-linked sugar chains, Examples include glycosaminoglycan, starch, amylose, amylopectin, cellulose, chitin, glycogen, agarose, alginic acid, hyaluronic acid, inulin, glucomannan and the like.
In the present invention, the sugar residue may include a modified sugar residue. In the present invention, a modified sugar is one that has been modified in the process of isolation and purification from a naturally occurring one, one that has been modified enzymatically, one that has been chemically modified, and biology including microorganisms. May be modified by conventional techniques, including modifications known in the field of glycoscience, such as hydrolysis, redox, esterification, acylation, amination, etherification, nitration, dehydration Modifications by reaction, glycosylation and the like may be included.

As the dehydrating condensing agent used in the method for producing a glycoside of the present invention, a haloformamidinium derivative can be suitably used, and typically, a haloformamidinium derivative represented by the general formula (2) is used. Is done. The haloformamidinium derivative (2) can be obtained by treating the corresponding urea derivative with an appropriate halogenating agent such as a chlorinating agent. Examples of the halogenating agent include phosgene, oxalyl chloride, phosphorus pentachloride, phosphorus trichloride, phosphorus oxychloride, and bromides corresponding thereto. Specific examples of the compound (2) include 2-chloro-1,3-dimethylimidazolinium chloride (DMC), 2-chloro-1,3-dimethylimidazolinium Hexafluorophosphate (2-chloro-1,3-dimethylimidazolinium hexafluorophosphate; CIP), 1- (chloro-1-pyrrolidinylmethylene) pyrrolidinium hexafluorophosphate, 1- (chloro-1-pyrrolidinylmethylene) ) Pyrrolidinium tetrafluoroborate, chloro-N, N, N ', N'-tetramethylformamidinium hexafluorophosphate, N, N, N', N'-tetramethyl-O- (N-succinimidyl) ) Uronium tetrafluoroborate, O- (benzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium tetrafluoroborate, O- (benzotriazol-1-yl) -N , N, N ', N'-Tetramethyluronium hexafluorophosphate, O- (7-aza Benzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium tetrafluoroborate, (4R, 5R) -2-chloro-1,3-dimethyl-4,5-diphenyl-1 -Imidazolinium chloride, (4S, 5S) -2-Chloro-1,3-dimethyl-4,5-diphenyl-1-imidazolinium chloride, N, N, N ', N'-tetramethylchloroformamid Nitrogen chloride (N, N, N ', N'-tetramethylchloroformamidinium chloride), Chloro-N, N, N', N'-bis (tetramethylene) formamidinium hexafluorophosphate (chloro-N, N, N ', N'-bis (tetramethylene) formamidinium hexafluorophosphate), 2-chloro-1,3-dimethyl-3,4,5,6-tetrahydro-2 (1H) -pyrimidinium chloride, and the like.

In practicing the present invention, glycosides can be produced for all sugars. The present invention can be practiced with any liquid in which the sugar dissolves, but preferably with water.
The synthesis reaction of compound (3) using compound (2) can be performed in a medium such as a solvent known in the art as long as the reaction is not adversely affected. The reaction may be in the absence of a solvent (including the case where the reaction raw material also serves as a solvent), but it is usually advantageous to perform in the presence of a solvent inert to the reaction. The solvent is not particularly limited as long as the reaction proceeds, but an aqueous solvent can be preferably used, such as water, methanol, ethanol, n-propanol, isopropanol, cyclohexanol, furfuryl alcohol, ethylene glycol, benzyl alcohol, and the like. Alcohols such as tetrahydrofuran (THF), dioxane, tetrahydrofurfuryl alcohol, diethylene glycol, cyclohexyl methyl ether, methyl cellosolve, cellosolve, butyl cellosolve, methyl tert-butanol and the like ethers such as methyl ethyl ketone, furfural, methyl isobutyl ketone, Ketones such as mesityl oxide, diacetone alcohol and cyclohexanone, for example, nitriles such as acetonitrile and benzonitrile, such as dimethyls Sulfoxides such as hydroxide (DMSO) and sulfolane, for example, amides such as formamide, N, N-dimethylformamide (DMF), and N, N-dimethylacetamide, such as methyl formate, ethyl formate, ethyl acetate, butyl acetate, Esters such as methoxybutyl acetate, cellosolve acetate, diethyl carbonate and glycol carbonate, for example, organic acids such as formic acid, acetic acid, propionic acid and acetic anhydride, heterocyclic compounds such as hexamethylphosphorotriamide, pyridine and quinoline, aniline, Aromatic amines such as N-methylaniline, nitro compounds and the like can be mentioned. These solvents can be used singly or, if necessary, two or more of them can be mixed at an appropriate ratio, for example, a ratio of 1: 1 to 1: 1000.

As the reaction medium for the reaction, water and a commonly used organic solvent can be used, but an aqueous solution is preferable, and an organic solvent containing water can also be used. In particular, an aqueous salt solution having an amine is more preferable. Moreover, the salt aqueous solution which has buffer capacity can also be used. The buffer can be selected from those known in the art as long as it does not adversely affect the reaction, and examples thereof include a phosphate buffer, a citrate buffer, and a Tris buffer. .
Typically, the hydrogen ion concentration pH exhibited by the amine solution is 1.0 to 13, more preferably 7.5 to 11. The reaction temperature is not particularly limited as long as the reaction proceeds,
Typically, it may be in a temperature range in which the reaction mixture does not solidify or boil, and in particular, it can be carried out in a temperature range in which the aqueous solution does not solidify or boil, and can be selected from 0 ° C. to 80 ° C., for example. Usually, it is desirable to carry out the reaction preferably in the range of 0 ° C to 50 ° C, more preferably at room temperature. In some cases, the reaction temperature can be preferably carried out at 0-10 ° C. The reaction time is not particularly limited and may be appropriately set as long as a desired product is obtained. For example, the reaction time may be 1 minute to 24 hours, and usually 15 minutes to 5 hours. In typical cases, it may be 15 minutes to 2 hours. The concentration of added sugar in the reaction system is preferably 1 mM to a saturated concentration, and more preferably 10 mM or more. Although the amount of the dehydrating condensing agent is not particularly limited, it can be suitably carried out at 1 to 5 equivalents, more preferably 3 equivalents with respect to the sugar. The amine concentration is 0.1 to 100 equivalents, more preferably 1 to 4 equivalents, based on the dehydration condensing agent used. For example, 6 to 9 equivalents of triethylamine is appropriate.

The amine may be a primary amine, a secondary amine, a tertiary amine, or a quaternary amine. For example, an aliphatic hydrocarbon residue, an aromatic hydrocarbon residue, a complex amine The aliphatic hydrocarbon residue may be linear or branched, and may be saturated or unsaturated, such as an alkyl residue. Group, alkenyl group, cycloalkyl group, aralkyl group, cycloalkylalkyl group, etc., and the aromatic hydrocarbon residue may be monocyclic or bicyclic or more condensed For example, a phenyl group, a naphthyl group, and the like can be mentioned, and the heterocyclic residue may have one or more selected from the group consisting of a sulfur atom, an oxygen atom, and a nitrogen atom, a pyridyl group, Imidazoli Group, a thiazolyl group, may like quinolinyl groups are included, as the relevant amine, piperidine, morpholine, thiomorpholine, piperazine, pyrrolidine, etc. may also be included. Representative examples of the amine include tertiary amines having aliphatic hydrocarbon residues such as trimethylamine, triethylamine, diethylmethylamine, dimethylethylamine, n-butyldimethylamine, diisopropylethylamine, tetramethylethylenediamine, and the like. Or diamines are mentioned.

The product can be used in the next reaction as the reaction solution or as a crude product, but can also be isolated from the reaction mixture according to a conventional method, and can be concentrated, concentrated under reduced pressure, distillation, fractional distillation, solvent extraction, liquid It can be isolated and purified by conversion, phase transfer, chromatography such as high performance liquid chromatography (HPLC), thin layer chromatography (TLC), column chromatography, crystallization, recrystallization and the like. For example, after completion of the reaction, the product can be extracted from the reaction solution with an organic solvent such as butyl acetate, ethyl acetate, toluene, chloroform, and the solvent can be distilled off to obtain a crude product. The crude product may be used as it is, but if necessary, after purification by means such as silica gel column chromatography, high performance liquid chromatography using a carrier such as a cellulose derivative (including an optically active carrier). It may be purified by such means as above. In an embodiment of the present invention, a method characterized by separating and / or purifying the obtained glycosylated glycoside product into a desired glucoside-containing glycoside by liquid chromatography can be constructed. In liquid chromatography, an ODS reverse phase column can be used to obtain a desired product efficiently and industrially advantageously.

Glycosides (3) obtained in the present invention include, for example, bioactive oligosaccharides, carriers of drug delivery systems, glycopharmaceuticals, surfactants, glycopeptides, glycoproteins, glycopolymers, etc. It is useful for various applications such as activity measurement reagents, surfactants, whitening agents, and enzyme-catalyzed sugar chain synthesis reagents.
The obtained glycoside, that is, a compound or oligosaccharide having a sugar chain added thereto is, for example, a bioactive oligosaccharide, a drug delivery system carrier, a surfactant, a sugar chain drug, a glycopeptide, a glycoprotein, a sugar chain height It is useful for various uses such as molecules. The product glycosides are useful for studies such as cell recognition, immunity, cell differentiation, cell migration, fertilization, maturation, tissue morphogenesis, inflammation, wound healing, cancer metastasis, tumorigenesis.
Among the glycosides (3) obtained in the present invention, for example, azide sugar can be used for known reactions including click reaction, Staudinger reaction and the like. For example, azide undergoes cycloaddition with various unsaturated bonds, for example, it adds to nitrile to form a tetrazole ring, and reacts with alkyne to form a 1,2,3-triazole ring. A sugar chain having an azide group can be immobilized on a solid phase such as filter paper or a carrier using a click reaction, and can be advantageously used for studying the interaction between a sugar chain and a biological substance. Furthermore, it is possible to incorporate azide sugar into cells and bind a fluorescent dye or the like bound to alkyne in vivo. Reaction of azide with phosphine (phosphite) produces iminophosphorane. Iminophosphorane is converted into amine and phosphine oxide by hydrolysis. Iminophosphorane also reacts with aldehydes to give imines. In addition, the sugar chain can be immobilized on a solid phase such as a gold substrate via a thiol to create a localized surface plasmon resonance device, which can be used to study interactions with lectins and the like.

Using the technology of the present invention, glycosides can be obtained by highly regioselective and / or stereoselective glycosylation, and can also be applied to long sugar chains, thus increasing the glycation variety. It is possible to introduce new oligosaccharide chains and / or polysaccharides into various organic compounds (including fluorescent compounds, luminescent compounds, etc.), peptides, proteins, lipids, carbohydrates, etc. Makes it possible to obtain glycosides. With the technology of the present invention, glycosides can be synthesized using oligosaccharide chains whose structure has been clarified, which is advantageous for application in various fields such as pharmaceuticals, agricultural chemicals and cosmetics. The present invention can provide a technique for producing a sugar chain microarray (sugar chain chip).
The present invention will be described in detail with reference to the following examples, which are provided merely for the purpose of illustrating the present invention and for reference to specific embodiments thereof. These exemplifications are for explaining specific specific embodiments of the present invention, but are not intended to limit or limit the scope of the invention disclosed in the present application. In the present invention, it should be understood that various embodiments based on the idea of the present specification are possible.
All examples were performed or can be performed using standard techniques, except as otherwise described in detail, and are well known and routine to those skilled in the art. .

(Synthesis of D-glucosyl azide)

To a solution of D-glucose (250 mM) in D ₂ O (0.5 ml), 10 equivalents of triethylamine and 10 equivalents of sodium azide were added to D-glucose. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to D-glucose were mixed at room temperature and stirred. After 30 minutes, sodium benzenesulfonate (10.0 mg, 0.056 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integral value of the anomeric proton of the produced azide sugar based on the integral value of the proton at the o-position of the standard substance (sodium benzenesulfonate), and was 94%. FIG. 1 shows the ¹ H NMR spectrum of the reaction solution containing the target product obtained in this example. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, Et ₃ N represents triethylamine, and PhSO ₃ Na represents sodium benzenesulfonate (the same applies hereinafter).

(Synthesis of D-galactosyl azide)

To a solution of D-galactose (250 mM) in D ₂ O (0.5 ml), 9 equivalents of triethylamine and 10 equivalents of sodium azide were added to D-galactose. Then 3 against D-galactose
It was mixed with an equivalent amount of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) at room temperature and stirred. After 30 minutes, sodium benzenesulfonate (10.0 mg, 0.056 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integrated value of the anomeric protons of the azide sugar produced based on the integrated value of the proton at the o-position of the standard substance (sodium benzenesulfonate), and was 67%. FIG. 2 shows the ¹ H NMR spectrum of the reaction solution containing the target product obtained in this example.

(Synthesis of D-mannosyl azide)

To a solution of D-mannose (250 mM) in D ₂ O (0.5 ml), 9 equivalents of triethylamine and 10 equivalents of sodium azide were added to D-mannose. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to D-mannose was mixed at room temperature and stirred. After 30 minutes, sodium benzenesulfonate (10.0 mg, 0.056 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integral value of the anomeric proton of the produced azide sugar based on the integral value of the proton at the o-position of the standard substance (sodium benzenesulfonate), and found to be 92%. FIG. 3 shows the ¹ H NMR spectrum of the reaction solution containing the target product obtained in this example.

(Synthesis of D-xylosyl azide)

To a solution of D-xylose (250 mM) in D ₂ O (0.5 ml), 9 equivalents of triethylamine and 10 equivalents of sodium azide were added to D-xylose. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to D-xylose were mixed at room temperature and stirred. After 30 minutes, sodium benzenesulfonate (10.0 mg, 0.056 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integrated value of the anomeric protons of the produced azide sugar based on the integrated value of the proton at the o-position of the standard substance (sodium benzenesulfonate), and was 97%. FIG. 4 shows the ¹ H NMR spectrum of the reaction solution containing the target product obtained in this example.

(Synthesis of D-cellobiosyl azide)

To a solution of D-cellobiose (250 mM) in D ₂ O (0.5 ml) was added 9 equivalents of triethylamine and 10 equivalents of sodium azide relative to D-cellobiose. Then 3 against D-cellobiose
It was mixed with an equivalent amount of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) at room temperature and stirred. After 30 minutes, sodium benzenesulfonate (10.0 mg, 0.056 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integrated value of the anomeric protons of the produced azide sugar based on the integrated value of the proton at the o-position of the standard substance (sodium benzenesulfonate), and found to be 86%. FIG. 5 shows the ¹ H NMR spectrum of the reaction solution containing the target product obtained in this example.

(Synthesis of D-maltosyl azide)

To a solution of D-maltose monohydrate (250 mM) in D ₂ O (0.5 ml), 9 equivalents of triethylamine and 10 equivalents of sodium azide were added to D-maltose. Then 3 against D-Maltose
It was mixed with an equivalent amount of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) at room temperature and stirred. After 30 minutes, sodium benzenesulfonate (10.0 mg, 0.056 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integral value of the anomeric proton of the produced azide sugar based on the proton integral value of the o-position of the standard substance (sodium benzenesulfonate), and was 80%. FIG. 6 shows the ¹ H NMR spectrum of the reaction solution containing the target product obtained in this example.

(Synthesis of D-lactosyl azide)

To a solution of β-D-lactose (250 mM) in D ₂ O (0.5 ml), 9 equivalents of triethylamine and 10 equivalents of sodium azide were added to D-lactose. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to D-lactose were mixed at room temperature and stirred. After 30 minutes, sodium benzenesulfonate (10.0 mg, 0.056 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integral value of the anomeric proton of the produced azide sugar based on the proton integral value at the o-position of the standard substance (sodium benzenesulfonate), and found to be 85%. FIG. 7 shows the ¹ H NMR spectrum of the reaction solution containing the target product obtained in this example.

(Synthesis of N-acetylglucosamino azide)

To a solution of N-acetylglucosamine (250 mM) in D ₂ O (0.5 ml), 7 equivalents of 2,6-lutidine and 10 equivalents of sodium azide were added to N-acetylglucosamine. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to N-acetylglucosamine was mixed at room temperature and stirred. After 24 hours, t-butyl alcohol (7.36 mg)
, 0.094 mmol) was added to the reaction solution, and ¹ H NMR was measured. The yield was calculated from the integral value of the anomeric proton of the produced azide sugar based on the proton integral value of the methyl group of the standard substance (t-butyl alcohol), and was 95%. FIG. 8 shows the ¹ H NMR spectrum of the reaction solution containing the target product obtained in this example.

(Synthesis of N-chitobiosyl azide)

To a solution of N-chitobiose (250 mM) in D ₂ O (0.5 ml), 6 equivalents of 2,6-
Llutidine and 10 equivalents of sodium azide were added. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to N-chitobiose were mixed at room temperature and stirred. After 24 hours, t-butyl alcohol (7.30 mg, 0.098 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integral value of the anomeric proton of the produced azide sugar based on the proton integral value of the methyl group of the standard substance (t-butyl alcohol), and was 94%. FIG. 9 shows the ¹ H NMR spectrum of the reaction solution containing the target product obtained in this example.

(Synthesis of glucuronic acid azide)

To a solution of sodium glucuronate (250 mM) in D ₂ O (0.5 ml) was added 15 equivalents of triethylamine and 10 equivalents of sodium azide relative to sodium glucuronate. Thereafter, 5 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to N-chitobiose was mixed at room temperature and stirred. After 30 minutes, t-butyl alcohol (7.30 mg, 0.098
mmol) was added to the reaction solution and ¹ H NMR was measured. Sodium benzenesulfonate (10.0 mg, 0.056 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. Yield is determined by reference material (
Based on the integral value of the proton at the o-position of sodium benzenesulfonate), it was calculated from the integral value of the anomeric proton of the produced azide sugar, which was 54%. FIG. 10 shows the ¹ H NMR spectrum of the reaction solution containing the target product obtained in this example.

(Synthesis of D-cellotriosyl azide)

To a solution of D-cellotriose (50 mM) in D ₂ O (0.5 ml), 9 equivalents of triethylamine and 10 equivalents of sodium azide were added to D-cellotriose. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to D-cellotriose were mixed at room temperature and stirred. After 30 minutes, sodium benzenesulfonate (5.0 mg, 0.028 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integrated value of the anomeric protons of the produced azide sugar based on the integrated value of the proton at the o-position of the standard substance (sodium benzenesulfonate), and was 30%. FIG. 11 shows the ¹ H NMR spectrum of the reaction solution containing the target product obtained in this example.

[Isolation of D-glucosyl azide]
D-glucose 112.5 mg (0.625 mmol) in D ₂ O (2.5 ml) was added 10 equivalents of triethylamine 867.5 μl (6.250 mmol) and 10 equivalents of sodium azide 406.5 mg (6.250 mmol) to D-glucose. In addition, it was cooled in ice water. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) 317.0 mg (1.875 mmol) with respect to D-glucose were mixed at 0 ° C. and stirred. After 30 minutes, the resulting reaction solution was reprecipitated with ethanol and silica gel column chromatography (developing solvent: acetonitrile / water = 5/1), HPLC (eluent: acetonitrile /
Purification by water = 3/1) gave the desired fraction. The obtained fraction was evaporated and freeze-dried to obtain 106.37 mg of β-D-glucosyl azide. The yield was 83%. FIG. 12 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of D-mannosyl azide]
To a solution of D-mannose 112.5 mg (0.625 mmol) in D ₂ O (2.5 ml), 10 equivalents of triethylamine 867.5 μl (6.250 mmol) and 10 equivalents of sodium azide 406.5 mg (6.250 mmol) were added to D-mannose. In addition, it was cooled in ice water. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) 317.0 mg (1.875 mmol) with respect to D-mannose was mixed at 0 ° C. and stirred. After 30 minutes, the resulting reaction solution was purified by reprecipitation with ethanol and silica gel column chromatography (developing solvent: chloroform / methanol = 5/1), HPLC (eluent: acetonitrile / water = 3/1). The desired fraction was obtained. The obtained fraction was evaporated and freeze-dried to obtain 103.9 mg of D-mannosyl azide. The yield was 81%. FIG. 13 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of 2-deoxyglucosyl azide]

To a solution of 2-deoxyglucose 40.8 mg (0.250 mmol) in D ₂ O (1.0 ml), 30 equivalents of triethylamine 1042-μl (7.50 mmol) and 20 equivalents of sodium azide 325.0 mg (5.00 mmol) ) And cooled in ice water. Then, 101.6 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) to 2-deoxyglucose 411.6 mg (2.50
mmol) at 0 ° C. and stirred. After 30 minutes, the resulting reaction solution was purified by reprecipitation with ethanol and silica gel column chromatography (developing solvent: chloroform / methanol = 8/1) to obtain the desired fraction. The obtained fraction was evaporated and lyophilized to obtain 38.5 mg of 2-deoxyglucopyranosyl azide. The yield was 82%. FIG. 14 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of N-acetylglucosamino azide]
To a solution of 138.25 mg (0.625 mmol) of N-acetylglucosamine in D ₂ O (1.0 ml) was added 434.3 μl (3.75 mmol) of 6 equivalents of 2,6-lutidine and 10 equivalents of sodium azide 406.3 with respect to N-acetylglucosamine. mg (62.5 mmol) was added. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) 317.0 mg (2.50 mmol) with respect to N-acetylglucosamine were mixed at 0 ° C. and stirred. After 24 minutes, the resulting reaction solution was purified by reprecipitation with ethanol and silica gel column chromatography (developing solvent: chloroform / methanol = 3/1) to obtain the desired fraction. The obtained fraction was evaporated and freeze-dried to obtain 64.3 mg of N-acetylglucosaminoazide. The yield was 70%. FIG. 15 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of D-cellobiosyl azide]
D-cellobiose 42.8 mg (0.125 mmol) in D ₂ O (0.5 ml) was charged with 9 equivalents of triethylamine 156 μl (1.125 mmol) and 10 equivalents of sodium azide 81.26 mg (1.250 mmol) with respect to D-cellobiose. added. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) 63.4 mg (0.375 mmol) with respect to D-cellobiose was mixed at room temperature and stirred. After 30 minutes, the resulting reaction solution was purified by reprecipitation with ethanol and HPLC (eluent: acetonitrile / water = 7/3) to obtain the desired fraction. The obtained fraction was evaporated and freeze-dried to obtain 33.6 mg of D-cellobiosyl azide. The yield was 73%. FIG. 16 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of D-maltosyl azide]
To a solution of D-maltose monohydrate 45.04 mg (0.125 mmol) in D ₂ O (0.5 ml), 9 equivalents of triethylamine 156 μl (1.125 mmol) and 10 equivalents of sodium azide 81.26 mg ( 1.250 mmol) was added. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) 63.4 mg (0.375 mmol) with respect to D-maltose were mixed at room temperature and stirred. After 30 minutes, the resulting reaction solution was purified by reprecipitation with ethanol and HPLC (eluent: acetonitrile / water = 7/3) to obtain the desired fraction. The obtained fraction was evaporated and freeze-dried to obtain 28.5 mg of D-maltosyl azide. The yield was 62%. FIG. 17 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of D-lactosyl azide]
D-Lactose 42.79 mg (0.125 mmol) in D ₂ O (0.5 ml) was charged with 9 equivalents of triethylamine 156 μl (1.125 mmol) and 10 equivalents of sodium azide 81.26 mg (1.250 mmol) to D-lactose. added. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) 63.4 mg (0.375 mmol) with respect to D-lactose were mixed at room temperature and stirred. After 30 minutes, the resulting reaction solution was purified by reprecipitation with ethanol and HPLC (eluent: acetonitrile / water = 7/3) to obtain the desired fraction. The obtained fraction was evaporated and freeze-dried to obtain 34.9 mg of D-lactosyl azide. The yield was 76%. FIG. 18 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of N-chitobiosyl azide]

To a solution of N-chitobiose 26.53 mg (0.0625 mmol) in D ₂ O (0.250 ml), 6 equivalents of 2,6-lutidine 43.4 μl (0.375 mmol) and 10 equivalents of sodium azide 40.63 mg ( 0.625 mmol) was added. Then, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) 31.7 mg (0.1875 mmol) with respect to N-chitobiose was mixed at room temperature and stirred. After 50 hours, the resulting reaction solution was purified by silica gel column chromatography (developing solvent: chloroform / methanol = 3/1) and HPLC (eluent: acetonitrile / water = 7/3). Obtained. The obtained fraction was evaporated and freeze-dried to obtain 13.26 mg of N-chitobiosyl azide. The yield was 49.4%. FIG. 19 shows the ¹ H NMR of the target product obtained in this example.
The spectrum is shown.

(Synthesis of N-chitotriosyl azide)

To a solution of N-chitotriose (50 mM) in D ₂ O (0.5 ml) was added 10 equivalents of 2,6-lutidine and 50 equivalents of sodium azide relative to N-chitotriose. Thereafter, 5 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to N-chitotriose was mixed at room temperature and stirred. After 36 hours, t-butyl alcohol (7.52 mg, 0.101 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integral value of the anomeric proton of the produced azide sugar based on the proton integral value of the methyl group of the standard substance (t-butyl alcohol) and found to be 75%. FIG. 20 shows the ¹ H NMR spectrum of the target product obtained in this example.

(Synthesis of N-chitotetraosyl azide)

To a solution of N-chitotetraose (20 mM) in D ₂ O (0.5 ml), 20 equivalents of 2,6-lutidine and 125 equivalents of sodium azide were added to N-chitotetraose. Thereafter, 10 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to N-chitotetraose was mixed at room temperature and stirred. After 36 hours, t-butyl alcohol (7.90 mg, 0.107 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integral value of the anomeric proton of the produced azide sugar based on the proton integral value of the methyl group of the standard substance (t-butyl alcohol), and was 69%. FIG. 21 shows the ¹ H NMR spectrum of the target product obtained in this example.

(Synthesis of D-maltotriosyl azide)

To a solution of D-maltotriose (50 mM) in D ₂ O (0.5 ml), 15 equivalents of N, N-diisopropylethylamine (DIPEA) and 50 equivalents of sodium azide were added to D-maltotriose. .
Then 5 equivalents of 2-chloro-1,3-dimethylimidazolinium to D-maltotriose
Mix with chloride (DMC) at room temperature and stir. After 30 minutes, sodium benzenesulfonate (10.0 mg, 0.056 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integral value of the anomeric proton of the produced azide sugar based on the integral value of the proton at the o-position of the standard substance (sodium benzenesulfonate), which was 74%. FIG. 22 shows the ¹ H NMR spectrum of the target product obtained in this example.

(Synthesis of D-maltotetraosyl azide)

To a solution of D-maltotetraose (50 mM) in D ₂ O (0.5 ml), 15 equivalents of N, N-diisopropylethylamine (DIPEA) and 50 equivalents of sodium azide were added to D-maltotetraose. . Thereafter, 5 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to D-maltotetraose was mixed at room temperature and stirred. After 30 minutes, sodium benzenesulfonate (5.0 mg, 0.028 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integral value of the anomeric proton of the azide sugar produced based on the integral value of the proton at the o-position of the standard substance (sodium benzenesulfonate), and was 71%. FIG.
FIG. 3 shows the ¹ H NMR spectrum of the target product obtained in this example.

(Synthesis of D-maltopentaosyl azide)

To a solution of D-maltopentaose (50 mM) in D ₂ O (0.5 ml), 15 equivalents of N, N-diisopropylethylamine (DIPEA) and 50 equivalents of sodium azide were added to D-maltopentaose. . Thereafter, 5 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to D-maltopentaose was mixed at room temperature and stirred. After 30 minutes, sodium benzenesulfonate (5.02 mg, 0.028 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integral value of the anomeric proton of the produced azide sugar based on the proton integral value at the o-position of the standard substance (sodium benzenesulfonate), and was 53%. FIG.
FIG. 4 shows the ¹ H NMR spectrum of the target product obtained in this example.

(Synthesis of D-maltohexaosyl azide)

To a solution of D-maltohexaose (50 mM) in D ₂ O (0.5 ml), 15 equivalents of N, N-diisopropylethylamine (DIPEA) and 50 equivalents of sodium azide were added to D-maltohexaose. . Thereafter, 5 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to D-maltohexaose was mixed at room temperature and stirred. After 30 minutes, sodium benzenesulfonate (5.04 mg, 0.028 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integral value of the anomeric proton of the produced azide sugar based on the proton integral value at the o-position of the standard substance (sodium benzenesulfonate), and was 53%. FIG.
5 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of D-maltotriosyl azide]
To a solution of 12.61 mg (0.025 mmol) of D-maltotriose in D ₂ O (0.5 ml), 15 equivalents of N, N-diisopropylethylamine (DIPEA) with D-maltotriose (65.3 μl, 0.375 mmol) and 50 An equivalent amount of sodium azide 81.3 mg (1.250 mmol) was added. Thereafter, 2 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) 21.1 mg (0.125 mmol) to D-maltotriose was mixed at room temperature and stirred. After 30 minutes, the resulting reaction solution was purified by HPLC (eluent: acetonitrile / water = 2/1) to obtain the desired fraction. The obtained fraction was evaporated and freeze-dried to obtain 10.3 mg of D-maltotriosyl azide. The yield was 78%. FIG.
FIG. 6 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of D-maltotetraosyl azide]
To a solution of D-maltotetraose 16.6 mg (0.025 mmol) in D ₂ O (0.5 ml), 15 equivalents of N, N-diisopropylethylamine (DIPEA) with D-maltotetraose 65.3 μl (0.375 mmol) and 50 An equivalent amount of sodium azide 81.3 mg (1.250 mmol) was added. Thereafter, 2 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) 21.1 mg (0.125 mmol) to D-maltotetraose was mixed at room temperature and stirred. After 30 minutes, the resulting reaction solution was purified by HPLC (eluent: acetonitrile / water = 2/1) to obtain the desired fraction. The obtained fraction was evaporated and freeze-dried to obtain 9.39 mg of D-maltotetraosyl azide. The yield was 55%. FIG. 27 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of D-maltopentaosyl azide]
A solution of 20.7 mg (0.025 mmol) of D-maltopentaose in D ₂ O (0.5 ml) was added with 15 equivalents of N, N-diisopropylethylamine (DIPEA) 65.3 μl (0.375 mmol) and 50 to D-maltopentaose. An equivalent amount of sodium azide 81.3 mg (1.250 mmol) was added. Thereafter, 2 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) 21.1 mg (0.125 mmol) to D-maltopentaose was mixed at room temperature and stirred. After 30 minutes, the resulting reaction solution was purified by HPLC (eluent: acetonitrile / water = 3/2) to obtain the desired fraction. The obtained fraction was evaporated and freeze-dried to obtain 15.38 mg of D-maltopentaosyl azide. The yield was 72%. FIG. 28 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of D-maltohexaosyl azide]
A solution of 12.39 mg (0.0125 mmol) of D-maltohexaose in D ₂ O (0.5 ml) was charged with 32.66 μl (0.1875 mmol) of N, N-diisopropylethylamine (DIPEA) 15 equivalents to D-maltohexaose. An equivalent amount of sodium azide 81.3 mg (1.250 mmol) was added. Thereafter, 5 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) (10.57 mg, 0.0625 mmol) with respect to D-maltohexaose were mixed at room temperature and stirred. After 30 minutes, the resulting reaction solution was purified by HPLC (eluent: acetonitrile / water = 3/2) to obtain the desired fraction. The obtained fraction was evaporated and freeze-dried to obtain 8.2 mg of D-maltohexaosyl azide. The yield was 65%. FIG. 29 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of D-Laminari Tetraosyl Azide]
D-laminaritetraose 16.7 mg (0.025 mmol) in D ₂ O (0.5 ml) was added to 15 equivalents of N, N-diisopropylethylamine (DIPEA) 65.3 μl (0.375 mmol) And 50 equivalents of sodium azide 81.3 mg (1.250 mmol) were added. Thereafter, 2 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) 21.1 mg (0.125 mmol) to D-laminaritetraose was mixed at room temperature and stirred. After 30 minutes, the resulting reaction solution was purified by HPLC (eluent: acetonitrile / water = 2/1) to obtain the desired fraction. The obtained fraction was evaporated and freeze-dried to obtain 11.2 mg of D-laminaritetraosyl azide. Yield 65
%Met. FIG. 30 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of D-Laminarihexaosyl Azide]
D-laminarihexaose (12.37 mg, 0.0125 mmol) in D ₂ O (0.5 ml) was added to 15 equivalents of N, N-diisopropylethylamine (DIPEA) 32.66 μl (0.1875 mmol) to D-laminarihexaose. And 100 equivalents of sodium azide 81.3 mg (1.250 mmol) were added. Thereafter, 5 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) (10.57 mg, 0.0625 mmol) to D-laminarihexaose was mixed at room temperature and stirred. After 30 minutes, the resulting reaction solution was purified by HPLC (eluent: acetonitrile / water = 3/2) to obtain the desired fraction. The obtained fraction was evaporated and freeze-dried to obtain 9.2 mg of D-laminarihexaosyl azide. The yield was 75%. FIG. 31 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of D-cellotetraosyl azide]

To a solution of D-cellotetraose 16.6 mg (0.025 mmol) in D ₂ O (0.5 ml), add 15 equivalents of N, N-diisopropylethylamine (DIPEA) 65.3 μl (0.375 mmol) and 50 to D-cellotetraose. An equivalent amount of sodium azide 81.3 mg (1.250 mmol) was added. Thereafter, 2 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) (21.1 mg, 0.125 mmol) to D-cellotetraose was mixed at room temperature and stirred. After 30 minutes, the resulting reaction solution was purified by HPLC (eluent: acetonitrile / water = 3/2) to obtain the desired fraction. The obtained fraction was evaporated and freeze-dried to obtain 14.12 mg of D-maltotetraosyl azide. The yield was 82%. FIG. 32 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of D-cellopentaosyl azide]

D-cellopentaose 10.3 mg (0.0125 mmol) in D ₂ O (0.5 ml) was added to D-cellopentaose 15 equivalents of N, N-diisopropylethylamine (DIPEA) 32.7 μl (0.188 mmol) and 100 An equivalent amount of sodium azide 81.3 mg (1.250 mmol) was added. Thereafter, 5 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) (10.57 mg, 0.0625 mmol) to D-cellopentaose was mixed at room temperature and stirred. After 30 minutes, the resulting reaction solution was purified by HPLC (eluent: acetonitrile / water = 3/2) to obtain the desired fraction. The obtained fraction was evaporated and freeze-dried to obtain 6.14 mg of D-cellopentaosyl azide. The yield was 70%. FIG. 33 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of D-cellohexaosyl azide]

To a solution of 4.95 mg (0.005 mmol) of D-cellohexaose in D ₂ O (0.5 ml), 15.01 μl (0.075 mmol) of N, N-diisopropylethylamine (DIPEA) with 15 equivalents of D-cellohexaose and 250 An equivalent amount of sodium azide 81.3 mg (1.250 mmol) was added. Thereafter, 5 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) 4.23 mg (0.025 mmol) with respect to D-cellohexaose was mixed at room temperature and stirred. After 30 minutes, the resulting reaction solution was purified by HPLC (eluent: acetonitrile / water = 3/2) to obtain the desired fraction. The obtained fraction was evaporated and freeze-dried to obtain 1.7 mg of D-cellohexaosyl azide. The yield was 34%. FIG. 34 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Isolation of (E, Z) 2-keto-3-penten-4-yl-β-D-glucopyranoside]
A solution of 225 mg (1.25 mmol) of glucose in H ₂ O (5 ml) was charged with 2.08 ml (15.0 mmol) of 12 equivalents of triethylamine and 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride ( DMC) 634 mg (3.75 mmol) was added. Thereafter, 1.93 ml (18.75 mmol) of 2,4-pentanedione equivalent to glucose was added, and the mixture was stirred at room temperature. After 3 hours, the resulting reaction solution was extracted with ethyl acetate. After recovering the aqueous layer, purification was performed by silica gel chromatography (developing solvent: chloroform / methanol = 9/1), and a fraction in which E-form and Z-form were mixed was collected. The obtained fraction was purified twice by HPLC (eluent: water 100%), and two fractions were collected. Each fraction was evaporated to 13.0 mg of E-2-keto-3-penten-4-yl-β-D-glucopyranoside and Z-2-keto-3-penten-4-yl-β-D-glucopyranoside 18.0 mg was obtained. The yield was 4.0% for E-form and 5.5% for Z-form. FIG. 35 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Synthesis of fluorinated β-glucopyranoside]
To a solution of glucose (250 mM) in D ₂ O (0.5 ml), 6 equivalents of triethylamine and 10 equivalents of potassium fluoride were added to glucose and cooled in ice water. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to glucose were mixed at 0 ° C. and stirred. After 15 minutes, sodium benzenesulfonate (9.0 mg, 0.050 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integral value of the anomeric protons of the produced fluorinated sugar, based on the proton integral value at the o-position of the standard substance (sodium benzenesulfonate), and was 23%. FIG. 36 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Synthesis of fluorinated β-glucopyranoside]
To a solution of glucose (250 mM) in D ₂ O (0.5 ml), 6 equivalents of triethylamine and 10 equivalents of cesium fluoride were added to glucose and cooled in ice water. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to glucose were mixed at 0 ° C. and stirred. After 15 minutes, sodium benzenesulfonate (9.0 mg, 0.050 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integral value of the anomeric protons of the produced fluorinated sugar, based on the proton integral value at the o-position of the standard substance (sodium benzenesulfonate), and was 12%. FIG. 37 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Synthesis of Fluorinated β-Glucopyranosyl]
To a solution of glucose (250 mM) in D ₂ O (0.5 ml), 6 equivalents of triethylamine and 10 equivalents of potassium fluoride were added to glucose and cooled in ice water. Then 3 equivalents of 2-chloro-1,3-dimethylimidazolinium hexafluorophosphate (CIP) to glucose
And stirred at 0 ° C. After 15 minutes, sodium benzenesulfonate (9.0 mg, 0.050 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integral value of the anomeric protons of the produced fluorinated sugar, based on the integral value of the proton at the o-position of the standard substance (sodium benzenesulfonate), and found to be 34%. FIG. 38 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Synthesis of fluorinated β-cellobiosyl]
To a solution of cellobiose (250 mM) in D ₂ O (0.5 ml), 6 equivalents of triethylamine and 10 equivalents of potassium fluoride were added to cellobiose and cooled in ice water. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to cellobiose were mixed at 0 ° C. and stirred. After 15 minutes, sodium benzenesulfonate (9.0 mg, 0.050 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integrated value of the anomeric protons of the produced fluorinated sugar, based on the integrated value of the proton at the o-position of the standard substance (sodium benzenesulfonate), and was 14%. FIG. 39 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Synthesis of Fluorinated β-Lactosyl]
To a solution of lactose (250 mM) in D ₂ O (0.5 ml), 6 equivalents of triethylamine and 10 equivalents of potassium fluoride were added to lactose and cooled in ice water. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to lactose were mixed at 0 ° C. and stirred. After 15 minutes, sodium benzenesulfonate (9.0 mg, 0.050 mmol) was added to the reaction solution as a standard substance, and ¹ H NMR was measured. The yield was calculated from the integrated value of the anomeric proton of the produced fluorinated sugar, based on the proton integrated value at the o-position of the standard substance (sodium benzenesulfonate), and was 19%. FIG. 40 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Synthesis of Fluorinated β-Lactosyl]
To a solution of lactose (250 mM) in D ₂ O (0.5 ml), 6 equivalents of triethylamine and 10 equivalents of potassium fluoride were added to lactose and cooled in ice water. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium hexafluorophosphate (CIP) with respect to lactose were mixed at 0 ° C. and stirred. After 15 minutes, sodium benzenesulfonate (9.0 mg, 0.050 mmol) was added to the reaction solution as a standard substance, and 1H NMR was measured. The yield was calculated from the integral value of the anomeric protons of the produced fluorinated sugar, based on the proton integral value at the o-position of the standard substance (sodium benzenesulfonate), and was 30%. FIG. 41 shows the ¹ H NMR spectrum of the target product obtained in this example.

[Synthesis of thioglucopyranoside]
To a mixed solution of glucose (250 mM) in D ₂ O (0.25 ml) and THF (0.25 ml), 9 equivalents of triethylamine and 10 equivalents of thiophenol were added to glucose. Thereafter, 3 equivalents of 2-chloro-1,3-dimethylimidazolinium chloride (DMC) with respect to glucose were mixed at room temperature and stirred. After 30 minutes, ¹ H NMR was measured. The yield was calculated from the integral value of the anomeric protons of the produced thioglucoside, assuming the synthesis of the integral value of the product and the remaining anomeric protons as 100%. 12%). FIG. 42 shows the ¹ H NMR spectrum of the target product obtained in this example.

In the present invention, a simple method for producing a glycoside from an unprotected sugar is provided. In the present invention, a glycoside is produced in one step in an aqueous solution from a sugar having a free hemiacetal hydroxy group using a formamidine derivative as a dehydrating condensing agent. Moreover, according to the production method of the present invention, production of a sugar chain probe used for a sugar chain chip can be easily achieved. The obtained glycosides, that is, glycoside-added compounds and oligosaccharide glycosides are exemplified by bioactive oligosaccharides, carriers of drug delivery systems, surfactants, sugar chain pharmaceuticals, glycopeptides, sugars, etc. The product glycoside is useful for various applications such as protein and sugar chain macromolecules, and the product glycoside is useful for cell recognition, immunity, cell differentiation, cell migration, fertilization, maturation, tissue morphogenesis, inflammation, wound It is useful for studies such as healing, cancer metastasis, and tumorigenesis.
It will be apparent that the invention may be practiced otherwise than as particularly described in the foregoing description and examples. Many modifications and variations of the present invention are possible in light of the above teachings, and thus are within the scope of the claims appended hereto.
It will be apparent that the invention may be practiced otherwise than as particularly described in the foregoing description and examples. Many modifications and variations of the present invention are possible in light of the above teachings, and thus are within the scope of the claims appended hereto.

1 is a ¹ H NMR spectrum of a reaction solution containing a target product obtained in Example 1. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, Et ₃ N represents triethylamine, and PhSO ₃ Na represents sodium benzenesulfonate. 2 is a ¹ H NMR spectrum of a reaction solution containing a target product obtained in Example 2. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, Et ₃ N represents triethylamine, and PhSO ₃ Na represents sodium benzenesulfonate. 2 is a ¹ H NMR spectrum of a reaction solution containing a target product, obtained in Example 3. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, Et ₃ N represents triethylamine, and PhSO ₃ Na represents sodium benzenesulfonate. 2 is a ¹ H NMR spectrum of a reaction solution containing a target product, obtained in Example 4. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, Et ₃ N represents triethylamine, and PhSO ₃ Na represents sodium benzenesulfonate. ^{1 is} a ¹ H NMR spectrum of a reaction solution containing a target product obtained in Example 5. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, Et ₃ N represents triethylamine, and PhSO ₃ Na represents sodium benzenesulfonate. 2 is a ¹ H NMR spectrum of a reaction solution containing a target product obtained in Example 6. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, Et ₃ N represents triethylamine, and PhSO ₃ Na represents sodium benzenesulfonate. 2 is a ¹ H NMR spectrum of a reaction solution containing a target product obtained in Example 7. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, Et ₃ N represents triethylamine, and PhSO ₃ Na represents sodium benzenesulfonate. 2 is a ¹ H NMR spectrum of a reaction solution containing a target product obtained in Example 8. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, 2,6-Lu represents 2,6-lutidine, and t-Bu represents t-butyl alcohol. ^{1 is} a ¹ H NMR spectrum of a reaction solution containing a target product obtained in Example 9. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, 2,6-Lu represents 2,6-lutidine, and t-Bu represents t-butyl alcohol. ^{1 is} a ¹ H NMR spectrum of a reaction solution containing a target product, obtained in Example 10. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, Et ₃ N represents triethylamine, and PhSO ₃ Na represents sodium benzenesulfonate. 2 is a ¹ H NMR spectrum of a reaction solution containing a target product, obtained in Example 11. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, Et ₃ N represents triethylamine, and PhSO ₃ Na represents sodium benzenesulfonate. 2 is a ¹ H NMR spectrum of the target product obtained in Example 12. 2 is a ¹ H NMR spectrum of the target product obtained in Example 13. ¹ is a ¹ H NMR spectrum of a target product obtained in Example 14. ¹ is a ¹ H NMR spectrum of a target product obtained in Example 15. 2 is a ¹ H NMR spectrum of the target product obtained in Example 16. 2 is a ¹ H NMR spectrum of the target product obtained in Example 17. ¹ is a ¹ H NMR spectrum of a target product obtained in Example 18. 2 is a ¹ H NMR spectrum of the target product obtained in Example 19. 2 is a ¹ H NMR spectrum of the target product obtained in Example 20. 2 is a ¹ H NMR spectrum of the target product obtained in Example 21. 2 is a ¹ H NMR spectrum of the target product obtained in Example 22. 2 is a ¹ H NMR spectrum of a target compound obtained in Example 23. 2 is a ¹ H NMR spectrum of a target compound obtained in Example 24. ¹ is a ¹ H NMR spectrum of a target product obtained in Example 25. 2 is a ¹ H NMR spectrum of a target product obtained in Example 26. 2 is a ¹ H NMR spectrum of a target compound obtained in Example 27. 2 is a ¹ H NMR spectrum of a target product obtained in Example 28. 2 is a ¹ H NMR spectrum of the target product obtained in Example 29. 2 is a ¹ H NMR spectrum of the target product obtained in Example 30. 2 is a ¹ H NMR spectrum of the target product obtained in Example 31. 2 is a ¹ H NMR spectrum of the target product obtained in Example 32. 2 is a ¹ H NMR spectrum of a target product obtained in Example 33. ¹ is a ¹ H NMR spectrum of a target compound obtained in Example 34. ^{1 is} a ¹ H NMR spectrum of a reaction solution containing a target product, obtained in Example 35. 2 is a ¹ H NMR spectrum of a reaction solution containing the target product, obtained in Example 36. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, Et ₃ N represents triethylamine, and PhSO ₃ Na represents sodium benzenesulfonate. 2 is a ¹ H NMR spectrum of a reaction solution containing the target product obtained in Example 37. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, Et ₃ N represents triethylamine, and PhSO ₃ Na represents sodium benzenesulfonate. 2 is a ¹ H NMR spectrum of a reaction solution containing the target product obtained in Example 38. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, Et ₃ N represents triethylamine, and PhSO ₃ Na represents sodium benzenesulfonate. 2 is a ¹ H NMR spectrum of a reaction solution containing the target product, obtained in Example 39. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, Et ₃ N represents triethylamine, and PhSO ₃ Na represents sodium benzenesulfonate. 2 is a ¹ H NMR spectrum of a reaction solution containing the target product, obtained in Example 40. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, Et ₃ N represents triethylamine, and PhSO ₃ Na represents sodium benzenesulfonate. 2 is a ¹ H NMR spectrum of a reaction solution containing the target product obtained in Example 41. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, and Et ₃ N represents triethylamine. 2 is a ¹ H NMR spectrum of a reaction solution containing the target product, obtained in Example 42. In the figure, DMI represents 1,3-dimethyl-2-imidazolidinone, and Et ₃ N represents triethylamine.

Claims (3)

General formula (1):

(In the above formula, R ¹ , R ² , R ³ and R ⁴ may be the same or different, and independently of each other, a hydrogen atom, a hydroxy group, a hydroxymethyl group, a carboxymethyl group, a carboalkoxymethyl group, amino group, an acylamino group including acetylamino group, a carboxyl group, an amide _{^{group, PO 4 2-, SO 4 -}} , alkyl group, acyl group, an alkoxy group, an acyloxy group, glucose, galactose, mannose, N- acetylglucosamine Selected from the group consisting of sugar residues of monosaccharides, sugar oligosaccharides, cellooligosaccharides, laminary oligosaccharides, chitooligosaccharides, xylooligosaccharides, and sugar residues of polysaccharides)
And an unprotected sugar represented by the general formula MZ (wherein Z is substituted with a hydrogen atom, a halogen atom, an azide group , a tetraazonium group, a nitro group, an optionally substituted hydrocarbon group or a hydrocarbon group) Oxy group, phosphoric acid residue, phosphonic acid residue, phosphinic acid residue, phosphorous acid residue, phosphonous acid residue, phosphinic acid residue, or substituted The hydrocarbon group which may be directly bonded to the phosphorus atom, or bonded to the phosphorus atom through an oxygen atom, a sulfur atom and / or a nitrogen atom, and phosphoric acid, phosphonic acid, phosphinic acid, phosphorous acid, those selected from nitrous acid residues phosphonic acid or derivative of phosphinous, and Ranaru group or an optionally substituted thio group with a hydrocarbon group, M is a metal, -OH, A compound of ammonium or hydrogen) General formula (2):

(In the above formula , R ⁶ and R ⁸ may be the same or different, and independently of each other, an optionally substituted alkyl group, an optionally substituted alkenyl group, and an optionally substituted aryl. those selected from the group consisting of groups forms a imidazolinium ^{R 5 -N-C + -N-} R 7 together, X is halogen atom, and Y ^- is an anion Is)
In an aqueous solution in the presence of an amine and a dehydrating condensing agent represented by the general formula (3):

(In the above formula, Z, R ¹ , R ² , R ³ and R ⁴ have the same meaning as above)
A method for producing a glycoside, comprising obtaining a glycoside represented by the formula: The dehydrating condensing agent represented by the general formula (2) is 2-chloro-1,3-dimethylimidazolinium chloride, 2-chloro-1,3-dimethylimidazolinium hexafluorophosphate , ( 4R, 5R) 2-chloro-1,3-dimethyl-4,5-diphenyl-1-imidazolinium chloride, and (4S, 5S) -2-chloro-1,3-dimethyl-4,5-diphenyl-1-imidazo The method for producing a glycoside according to claim 1, wherein the glycoside is selected from the group consisting of linium chloride. General formula (3) (wherein Z is selected from the group consisting of an azide group, a halogen atom, and an arylthio group, and R ¹ , R ² , R ³ and R ⁴ have the same meaning as described above. The method for producing a glycoside according to claim 1 or 2, wherein the glycoside represented by (1) is obtained.

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