WO2021054474A1

WO2021054474A1 - Repeating disaccharide for oligosaccharide synthesis and method for producing oligomer thereof

Info

Publication number: WO2021054474A1
Application number: PCT/JP2020/035624
Authority: WO
Inventors: 田村　純一; 敬裕田村
Original assignee: 国立大学法人鳥取大学
Priority date: 2019-09-20
Filing date: 2020-09-18
Publication date: 2021-03-25
Also published as: JPWO2021054474A1

Abstract

In the present invention, a disaccharide unit for synthesizing a useful sugar chain for suppressing the development of dystroglycanopathy is obtained and used to reconstitute a post-phosphoryl sugar chain.　Provided are: a disaccharide derivative represented by formula (1) [wherein X¹ is a leaving group or a protected hydroxyl group, Y¹ is a carboxyl group, a hydroxymethyl group, or a precursor group thereof, and R¹ to R⁵ are independently hydrogen or a protecting group for a hydroxyl group]; a production method for the same; and a method for producing an oligomer of the disaccharide derivative using the disaccharide derivative as an intermediate.

Description

Repetitive disaccharides involved in oligosaccharide synthesis and methods for producing their oligomers

The present invention relates to a synthetic intermediate for sugar chains and a method for producing the same.

Abnormal muscular dystrophy, which is a type of muscular dystrophy, is caused by abnormal biosynthesis of dystroglycan, which is a glycoprotein, and is accompanied by central nervous system disorders such as brain malformations and mental retardation, including muscle cell damage. Recent studies have revealed that dystroglycan sugar chain abnormalities present in muscle cells are the cause of sugar chain abnormal muscular dystrophy. Of the sugar chains of dystroglycan, the sugar chain portion that has an affinity for laminin is called matriglican. Matriglycan binds muscle cells to the basement membrane via laminin, and abnormal biosynthesis occurs in this sugar chain, resulting in the development of abnormal sugar chain muscular dystrophy.

Research has been carried out on the overall structure of sugar chains after Ser / Thr containing matriglycan, and it is known that it is an O-mannosylglycan in which matriglican is elongated via ribitol phosphate. Matriglican contains a sugar chain having a repeating disaccharide consisting of α-xylose and β-glucuronic acid at its non-reducing terminal portion (Non-Patent Document 1), and the sugar chain having repeating this disaccharide is laminin. It is known that it is necessary for the binding of (Non-Patent Document 2).

For the treatment of sugar chain abnormal muscular dystrophy, improvement of the gene that regulates the sugar chain biosynthesis enzyme involved in the biosynthesis of O-mannosyl glycan is expected. However, there are many types of genes involved, and it has been reported that the condition worsens when overexpressed, and gene therapy for sugar chain abnormal muscular dystrophy is not easy.

If the matrix on the dystroglycan that connects the muscle cells and the basement membrane via laminin can be reconstructed, the onset of sugar chain abnormal muscular dystrophy can be suppressed. Therefore, the problem to be solved by the present invention is to reconstruct the matrix glycan.

The present inventors have conducted intensive studies to solve the above problems, and synthesized a disaccharide unit (Xylα1-3GlcAβ) for synthesizing a sugar chain having a repeating disaccharide consisting of α-xylose and β-glucuronic acid. , Succeeded in the synthesis of tetrasaccharides in which the disaccharide units are bound to each other, and the synthesis of the tetrasaccharide moiety that connects the sugar chain having the repetition and the sugar chain extending from the α-dystroglycan, and showed the possibility of reconstructing the matrix. It was. The present inventors have completed the present invention from these findings.

Therefore, the present invention provides the following.
[1] Equation (1):

[In the formula, X ¹ is a leaving group or a protected hydroxyl group, Y ¹ is a carboxyl group or a hydroxymethyl group or a group that is a precursor thereof, and R ¹ to R ⁵ are independently hydrogen. Is or is a hydroxyl-protecting group]
A disaccharide derivative represented by.

[2] X ¹ is a group selected from the group consisting of trichloroacetimideyloxy, alkylthio, arylthio, halogen, pentenyloxy, alkoxy and phenoxy, Y ¹ is COOZ ¹ or CH ₂ OZ ² , and Z ¹ Is an alkyl group, Z ² is a protecting group for hydroxyl groups, and R ¹ to R ⁵ are independently substituted benzyl-containing benzyl, allyl, levulinoyl, substituted benzoyl-containing benzoyl, substituted acetyl-containing acetyl, and allyloxy. The disaccharide derivative according to [1], which is a group selected from the group consisting of carbonyl, isopropylidene, benzylidene and trialkylsilyl.

[3] Equation (2):

[In the formula, X ² is a leaving group, Y is a group that fixes the configuration of the xylose residue, and R ⁶ is immobilized with p-alkyloxybenzyl, 3,4-dialkyloxybenzyl. P-alkyloxybenzyl, immobilized 3,4-dialkyloxybenzyl, naphthyl, naphthylmethyl, -CH = CH ₂ , -C (CH ₃ ) = CH ₂ , -CH ₂ -CH = CH ₂ , -CH = CH-CH ₃ , -CH = C = CH ₂ , and a group selected from the group consisting of hydrogen]
With the xylose derivative represented by
Equation (3):

[In the formula, X ³ is a leaving group or a protected hydroxyl group, R ⁷ and R ⁸ are a protecting group for a hydroxyl group, and Y ⁴ is a group that is a precursor of a carboxyl group or a hydroxymethyl group].
From the glucose derivative or glucuronic acid derivative indicated by
Equation (4):

[In the equation, X ³ , R ⁷ , R ⁸ and Y ⁴ are the same as the above definitions]
A method for producing an α-glycoside represented by the following steps 1) to 3) :.
Step 1) By reacting the xylose derivative represented by the formula (2) with the glucose derivative or the glucuronic acid derivative represented by the formula (3), the OR ⁶ group of the xylose derivative and the glucose derivative or the glucuronic acid derivative are not protected. Manufacture mixed acetal derivatives via hydroxyl groups
Step 2) mixed acetal derivative obtained with step 1) is reacted to produce [alpha] 1 → 3 disaccharide derivative by activating the leaving group X ^2, then,
Step 3) A method comprising deprotecting the disaccharide derivative obtained in Step 2) and inducing it into the formula (4).

[4] X ² is a group selected from the group consisting of trichloroacetimideyloxy, alkylthio, halogen, arylthio, and pentenyloxy, and Y is selected from the group consisting of acetal, carbonate, silylene acetal, and stanilen acetal. R ⁶ is a group selected from the group consisting of p-methoxybenzyl and naphthylmethyl, and X ³ is composed of trichloroacetimideyloxy, alkylthio, arylthio, halogen, pentenyloxy, alkoxy and phenoxy. A group selected from the group, each of which contains R ⁷ and R ⁸ independently of a substituted benzyl containing a benzyl, an allyl, a lebrinoyl, a substituted benzoyl containing a benzoyl, a substituted acetyl containing an acetyl, an allyloxycarbonyl and a trialkylsilyl. The method according to [3], wherein Y ⁴ is a COOZ ² or CH ₂ OZ ³ , Z ² is an alkyl group, and Z ^{3 is a hydroxyl group protective group.}

[5] The method according to [4], wherein R ⁷ and R ⁸ are groups selected from the group consisting of a benzoyl containing a substituted benzoyl, an acetyl containing a substituted acetyl and a lebrinoyl.

[6] Equation (2'):

Wherein, X 2 ^'is a leaving group, Y' is a group for fixing the conformation of xylose residues, R 6 ^'is a protecting group with no neighboring group participation ability hydroxyl]
With the xylose derivative represented by
Equation (3'):

Wherein, X 3 ^'is a leaving group or a protected hydroxyl group, R ^7' is hydroxyl-protecting group with a neighboring group participation ability, R 8 ^'is a protecting group for a hydroxyl group, Y ^4' represents a carboxyl A group that is a precursor of a group or a hydroxymethyl group]
From the glucose derivative or glucuronic acid derivative indicated by
Equation (4'):

^{^{Wherein, X 3 ', R 7'}} , R 8 ' and ^{Y 4'} are the same as defined above
A method for producing an α-glycoside represented by the following steps 1) to 3) :.
Step 1) The xylose derivative represented by the formula (2') and the glucose derivative or the glucuronic acid derivative represented by the formula (3') are directly condensed to obtain a mixture of α1 → 3 glycoside and β1 → 3 glycoside. ,
Step 2) Deprotect the mixture obtained in Step 1) and then
Step 3) A method comprising separating the α-glycoside from the deprotected mixture in Step 2).

[7] The method according to [6], wherein the condensation in step 1) is carried out using NIS-AgOTf, NIS-TfOH, MeOTf, CuBr ₂ -AgOTf-nBu ₄ NI, or NBS-AgOTf as a condensing agent.

[8] The deprotection in step 2) is to form triol having hydroxyl groups at the 2-, 3- and 4-positions of the xylose residue by hydrolysis using trifluoroacetic acid (TFA) [6]. ] Or the method according to [7].

[9] The method according to any one of [6] to [8], wherein the separation in step 3) is carried out by dissolving β-glycoside in chloroform to remove β-glycoside.

[10] The formula (5): including the use of the disaccharide derivative represented by the formula (1) as an intermediate (referred to as an XG unit):

[In the formula, X ⁴ is a leaving group or a protected hydroxyl group, and n is an integer greater than or equal to 1.]
A method for producing an XG unit oligomer represented by.

[11] The disaccharide derivative represented by the formula (1) is used as a sugar donor, and the formula (1'):

[In the formula, Y ⁵ is a group that is a precursor of a carboxyl group or a hydroxymethyl group, X ⁵ is a leaving group or a protected hydroxyl group, and R ⁹ to R ¹² are a protecting group of a hydroxyl group].
[10] The method according to [10], which comprises reacting the disaccharide derivative represented by (1) as a sugar receptor.

[12] Y ⁵ is COOZ ³ or CH ₂ OZ ⁴ , Z ³ is an alkyl group, Z ⁴ is a hydroxyl group protective group, and X ⁵ is trichloroacetimideyloxy, alkylthio, arylthio, halogen, pentenyl. It is a group selected from the group consisting of oxy, alkoxy and phenoxy, and R ⁹ to R ¹² are independently benzyl containing a substituted benzyl, allyl, levulinoyl, benzoyl containing a substituted benzoyl, acetyl containing a substituted acetyl, and allyl. The method according to [11], which is a group selected from the group consisting of oxycarbonyl and trialkylsilyl.

[13] R ⁹ is a group selected from the group consisting of acetyl and levulinoyl including benzoyl including substituted benzoyl, substituted acetyl [12] The method according.

[14] Equation (9):

Tetrasaccharides or derivatives thereof.

[15] Equation (7):

[In the formula, X ¹ is a leaving group or a protected hydroxyl group, X ² to X ⁶ are hydroxyl protecting groups, and X ⁷ is an alkyl group].
The disaccharide donor represented by the formula (8):

^Wherein, X 8 ^{~ X 13} is a hydroxyl-protecting group, nice contrast ^{X 12} and ^{X 13} may form acetal together]
The tetrasaccharide derivative is obtained by reacting with the disaccharide receptor represented by, and then the tetrasaccharide derivative is deprotected to obtain the formula (9) :.

A method for producing a tetrasaccharide represented by the formula (9), which comprises obtaining the tetrasaccharide represented by the formula (9).

[16] X ¹ is trichloroacetimideyloxy, X ² to X ⁶ are acetyl, X ⁷ is methyl, X ⁸ is allyl, X ⁹ and X ¹⁰ are benzyl, X ¹¹ is TBDPS, and X ¹² and X ¹³ are together. [15]. The method according to [15], wherein the isopropyrine acetal is formed.

According to the present invention, a disaccharide unit consisting of α-xylose, which may be derivatized, and β-glucuronic acid, which may be derivatized, is provided. The disaccharide unit can be used to reconstruct the postphosphate sugar chain. This makes it possible to suppress the onset of sugar chain abnormal muscular dystrophy.

1. 1. Disaccharide Derivatives In one embodiment, the present invention has the formula (1):

[In the formula, X ¹ is a leaving group or a protected hydroxyl group, Y ¹ is a carboxyl group or a hydroxymethyl group or a group that is a precursor thereof, and R ¹ to R ⁵ are independently hydrogen. Is or is a hydroxyl-protecting group]
The disaccharide derivative represented by is provided.

A ^{known leaving group or protected hydroxyl group can be used as X 1, and} it is generally used for sugar chain synthesis such as trichloroacetimideyloxy, alkylthio, arylthio, halogen, and pentenyloxy (4-pentenyloxy). Leaving groups are exemplified, but not limited to these. The same applies to leaving groups at other points in the present disclosure. Preferred leaving groups X ¹ include, but are not limited to, those that form thioglycosides such as trichloroacetimideyloxy, alkylthio and phenylthio. Hydroxyl protecting groups are well known to those of skill in the art. That is, protected hydroxyl groups are well known to those of skill in the art. X ¹ may be alkoxy or phenoxy.

Y ¹ is a carboxyl group or a hydroxymethyl group or a group which is a precursor thereof. Groups that are precursors of carboxyl or hydroxymethyl groups are well known to those skilled in the art and are typically carboxyled in one step through reactions such as oxidation / reduction reactions, hydrolysis reactions, heating and the like. Represents a group that can be converted to a group or a hydroxymethyl group. Precursors of carboxyl or hydroxymethyl groups are also usually derivatives of carboxyl or hydroxymethyl groups. ^{Examples of one} Y unit include, but are not limited to ^{, COOZ 1} or CH ₂ OZ ^2. Here, Z ¹ is an alkyl group, preferably an alkyl group having 1 to 3 carbon atoms. A more preferred Z ¹ group is methyl. Z ² is a commonly used hydroxyl protecting group. Hydroxyl protecting groups are known and specific examples thereof are described herein.

R ¹ to R ⁵ are each independently hydrogen or a hydroxyl-protecting group. The protecting group in the present disclosure is preferably a group that can be removed after sugar chain formation. In the present disclosure, when a compound contains a plurality of hydroxyl protecting groups, the plurality of protecting groups may be of different types from each other or may be of the same type as each other. Selective deprotection can be facilitated by using different types of protecting groups. Such hydroxyl protecting groups are known and include, for example, the protecting groups described in Chapter 2 of Greene's Protective Groups in Organic Synthesis, 5th Edition (Wiley). Specifically, a benzyl group, a p-methoxybenzyl group, a p-methoxyphenyl group, a p-nitrobenzyl group, a benzoyl group, a p-methylbenzoyl group, a benzylidene group, an acetyl group, a pivaloyl group, a lebrinoyl group, an allyl group, Examples thereof include, but are not limited to, a methoxymethyl group, a tert-butyldimethylsilyl group, a triisopropylsilyl group, an isopropylidene, a benzylidene, an allyloxycarbonyl, and a trialkylsilyl. Preferred R ¹ groups, benzoyl including substituted benzoyl with neighboring group participation ability, acetyl including substituted acetyl, including but levulinoyl, without limitation. Additional examples of protecting groups which neighboring group participation ability has been described for R ^{7 'group,} which will be described later, they can be used in ¹ group R. Preferred R ^{2 ~} R ⁵ groups, benzyl including substituted benzyl, allyl, levulinoyl, benzoyl, including substituted benzoyl, acetyl including substituted acetyl, allyloxycarbonyl, commonly used in the sugar chain synthesis such as trialkylsilyl Protecting groups include, but are not limited to. At least one of ^{R 1} to R ⁵ (eg, either ^{R 1} ^{or R 2} to R ⁵ ) is preferably a protecting group, in which case the remaining group is hydrogen. All of R ¹ to R ⁵ may be protecting groups.

In a further aspect of the present invention, the formula (2):

[In the formula, X ² is a leaving group, Y is a group that fixes the coordination of xylose residues, and R ⁶ is p-alkyloxybenzyl, 3,4-dialkyloxybenzyl, immobilized. p-alkyloxybenzyl, immobilized 3,4-dialkyloxybenzyl, naphthyl, naphthylmethyl, -CH = CH ₂ , -C (CH ₃ ) = CH ₂ , -CH ₂ -CH = CH ₂ ,- A group selected from the group consisting of CH = CH-CH ₃ , -CH = C = CH _{2, and hydrogen]}
With the xylose derivative represented by
Equation (3):

[In the equation, X ³ , R ⁷ , R ⁸ and Y ⁴ are the same as the above definitions]
A method for producing an α-glycoside represented by the following steps 1) to 3) :.
Step 1) By reacting the xylose derivative represented by the formula (2) with the glucose derivative or the glucuronic acid derivative represented by the formula (3), the OR ⁶ group of the xylose derivative and the glucose derivative or the glucuronic acid derivative are not protected. Manufacture mixed acetal derivatives via hydroxyl groups
Step 2) mixed acetal derivative obtained with step 1) is reacted to produce [alpha] 1 → 3 disaccharide derivative by activating the leaving group X ^2, then,
Step 3) Provided is a method comprising deprotecting the disaccharide derivative obtained in Step 2) and inducing it into the formula (4).

In the above method for producing an α-glycoside, the compound of the formula (2) is a sugar donor and the compound of the formula (3) is a sugar receptor.

In the compound of formula (2), X ² is a leaving group. X ² is the same as the leaving group X ¹ described above, and is a leaving group commonly used for sugar chain synthesis such as trichloroacetimideyloxy, alkylthio, arylthio, phenylsulfinyl, halogen, pentenyloxy and the like. Is exemplified, but the present invention is not limited to these. Trichloroacetimidate yl oxy Preferred X ² as leaving groups, but which forms a thioglycoside such as alkylthio or arylthio is exemplified, without limitation. The alkylthio group described as a leaving group in the present disclosure may be, for example, an alkylthio having 1 to 20 carbon atoms, but is not limited thereto. The arylthio group described as a leaving group in the present disclosure can be, but is not limited to, for example, phenylthio or trilthio.

In the sugar donor of formula (2), Y is a group capable of immobilizing the xylose residue in the notation conformation and preventing ring inversion. In the formula (2), the two oxygen atoms displayed on the left side are included in the Y group. Specifically, Y can be an acetal, carbonate, silylene acetal, or stanilen acetal. Acetals include diacetals. It should be understood that the oxygen atoms that define these structures correspond to the two oxygen atoms displayed on the left side of equation (2) above. More specifically, Y may have a structure represented by the following formula (2a).

In formula (2a), Z is C, Si, or Sn, and when Z = C and Y is an acetal, n is an integer of 0 to 2 (preferably 0 or 1), with the exception of this. In the case, n is 0. The two Ls attached to each Z are independently alkyl, phenyl, or hydrogen. It is preferred that either or both of the two L's are alkyl or phenyl. When n is 1 or more, one of the two Ls attached to each Z can be further alkyloxy (eg, methoxy) or phenyloxy. Alternatively, when Z = C and Y is carbonate, the two Ls bound to Z together represent "O =". Alkyl groups in alkyl and alkyloxy can be, for example, alkyl groups having 1 to 6 carbon atoms, but are not limited thereto.
Preferred specific examples of the sugar donor of the formula (2) are shown below. In the examples below, (2-1) and (2-2) are embodiments in which Y is an acetal with n = 0, and (2-3) is an embodiment in which Y is an acetal with n = 1, (2-). 4), (2-5), and (2-6) represent embodiments in which Y is carbonate, silylene acetal, and stanilen acetal, respectively. The L group shown here is an example, and the L group is not limited thereto.

R ⁶ is p-alkyloxybenzyl, 3,4-dialkyloxybenzyl, immobilized p-alkyloxybenzyl, immobilized 3,4-dialkyloxybenzyl, naphthyl, naphthylmethyl, -CH = It is selected from the group consisting of CH ₂ , -C (CH ₃ ) = CH ₂ , -CH ₂ -CH = CH ₂ , -CH = CH-CH ₃ , -CH = C = CH _{2, and hydrogen.} These R ⁶ are, IAD: 2-position of the sugar donor by (intramolecular aglycon delivery intramolecular aglycon transfer) method and the 3-position of the glycosyl acceptor (the only unprotected hydroxyl group) group to produce a mixed acetal connecting between the Is. The IAD method itself is known to those of skill in the art and is reviewed by Cumspstey in Carbohydrate Research, 343, 1553-1573 (2008). For example, R ⁶ is p-alkyloxybenzyl, 3,4-dialkyloxybenzyl, immobilized p-alkyloxybenzyl, immobilized 3,4-dialkyloxybenzyl, naphthyl, or naphthylmethyl. In the case of oxidative conditions using 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ), between the sugar receptor represented by the formula (3) (the third position of the sugar receptor). It gives rise to mixed acetals (via unprotected hydroxyl groups). As is understood by those skilled in the art, in addition to the DDQ, according to the type of R ^6, different reagents to produce a mixed acetal may be used (e.g., acid catalysts such as tosylate, N- Examples include, but are not limited to, iodosuccinimide (NIS) and base catalysts such as, but not limited to, imidazole and 4-dimethylaminopyridine (DMAP). In particular, when R ⁶ _{is hydrogen, Me 2} SiCl ₂ can be added to the reaction with the sugar receptor to participate in the formation of a silylene mixed acetal. As described in the above Cumpstey review, specific reagents for producing mixed acetals by the IAD method are known to those skilled in the art and can be appropriately selected by those skilled in the art. Alkyl group in the R ⁶ groups described above may for example those having 1 to 6 carbon atoms but not limited thereto. Preferred R ⁶ p-methoxybenzyl, naphthylmethyl and the like, without limitation.

In the sugar receptor of formula (3), X ³ is a leaving group or a protected hydroxyl group, and R ⁷ and R ⁸ are hydroxyl protecting groups. X ³ ^{is similar to X 1} described above, and R ⁷ and R ⁸ are similar to R ¹ to R ⁵ described above, especially R ¹ and R ² . Preferred X ³ as a leaving group, trichloroacetimidate benzyloxy, alkylthio, arylthio, halogen, such as pentenyloxy, leaving group commonly used in the sugar chain synthesis. Preferred R ⁷ and R ⁸ are generally used for the synthesis of sugar chains such as benzyl containing substituted benzyl, allyl, lebrinoyl, benzoyl containing substituted benzoyl, acetyl containing substituted acetyl, allyloxycarbonyl, trialkylsilyl and the like. Protecting groups can be mentioned. Particularly preferred R ^7, benzoyl including substituted benzoyl with neighboring group participation ability, acetyl including substituted acetyl, although such levulinoyl include, but are not limited to. Additional examples of protecting groups which neighboring group participation ability has been described for R ^{7 'group,} which will be described later, they may be used in the R ⁷ groups.

Y ⁴ ^{is similar to Y 1} described above, and examples thereof include, but are not limited to, ^{COOZ 2} or CH ₂ OZ ^3. Here, Z ² is an alkyl group, preferably an alkyl group having 1 to 3 carbon atoms. More preferred Z ² groups are methyl. Z ³ is a commonly used hydroxyl protecting group. Hydroxy group protecting groups are known.

The xylose derivative (sugar donor) represented by the formula (2) is condensed with the glucose derivative or glucuronic acid derivative (sugar receptor) represented by the formula (3) to obtain the α-glycoside represented by the formula (4). The steps for obtaining the α-glycoside include the following 1) to 3). Steps 1) to 3), particularly steps 1) and 2), are based on the IAD method.
Step 1) The xylose derivative represented by the formula (2) is reacted with the glucose derivative or the glucuronic acid derivative represented by the formula (3), and the OR ⁶ group at the 2-position of the xylose derivative is reacted with the glucose derivative or the glucuronic acid derivative. A mixed acetal derivative via an unprotected hydroxyl group at the 3-position of
Step 2) Step 1 mixed acetal derivative obtained with) were reacted to produce [alpha] 1 → 3 disaccharide derivative by causing activation of leaving groups X ^2, then,
Step 3) The disaccharide derivative obtained in Step 2) is deprotected and induced into the formula (4).

The reaction of step 1) can be carried out, for example, in the presence of 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ). The mixed acetal derivative produced in step 1) is a mixed acetal in which the 2-position carbon of the sugar donor and the 3-position carbon of the sugar receptor are bonded to two oxygen atoms of the acetal, respectively. The disaccharide derivative obtained in step 2) is a disaccharide in which the 1-position carbon of the sugar donor and the 3-position carbon of the sugar receptor are linked to the same one oxygen atom, as in the formula (4). is there. Different types of various condensing agent leaving group X ² can activate (glycosylation promoter) it is known to those skilled in the art and are described in more detail in Cumpstey review as part of the IAD process. The reaction of step 2) can be carried out in the presence of a condensing agent (glycosylation promoter) such as NIS-AgOTf, NIS-TfOH, MeOTf, CuBr ₂ -AgOTf-nBu _{4 NI, NBS-AgOTf.} Step 3) is a deprotection step. Although the deprotection method differs depending on the type of protecting group, various deprotecting methods of the protecting group are known. For example, in the case of a benzyl group or the like, deprotection may be performed by hydrolysis with an acid or alkali. Trifluoroacetic acid is particularly preferably used for deprotection. The Y group is also removed by deprotection, and a hydroxyl group can be left at the 3rd and 4th positions of xylose.

A specific example of the above method for producing an α-glycoside is shown in Scheme 1 below.

[In the above scheme, Lev is -C (= O) C ₂ H ₄ C (= O) CH ₃ , Bz is -C (= O) C ₆ H ₅ , and MBn is -CH ₂ C ₆ H ₄ OCH ₃ (p), MP is -C ₆ H ₄ OCH ₃ (p), Me is -CH ₃ , Ac is -C (= O) CH ₃ , and MBz is -C. (= O) C ₆ H ₄ CH ₃ (p)]

In a further aspect of the present invention, the xylose derivative represented by the above formula (2) and the glucose derivative or the glucuronic acid derivative represented by the above formula (3) are combined with 2,3-dichloro-5,6-dicyano-p. -A disaccharide comprising reacting in the presence of benzoquinone (DDQ) or the like to obtain a mixed acetal derivative, condensing the mixed acetal derivative in the presence of a condensing agent, and then deprotecting the condensate, if desired. (Xylose α1-3 glucuronic acid, in the disaccharide, the xylose residue and the glucuronic acid residue may be derivatized).
Deprotection, if desired, means removing some (at least one) or all of the protecting groups. Derivatization of sugars is known to those of skill in the art. For example, the disaccharide may contain a known protecting group or may contain a known leaving group. Examples of the disaccharide protecting group and leaving group include those described above, but are not limited thereto.

The reaction between the xylose derivative (sugar donor) corresponding to the formula (2) and the glucose derivative or the glucuronic acid derivative (sugar receptor) corresponding to the formula (3) had the following problems:
-Since adjacent group involvement cannot be used for α-xylose, it is difficult to control the anomeric position.
-Xylose is easy to flip the ring.
Due to these problems, α-glycosidic bonds may not be formed.
Therefore, the present inventors protect the hydroxyl group at the 2-position of xylose with a protecting group that causes an intramolecular aglycon transfer and condense it with a glycosyl acceptor (glucuronic acid derivative) to cause an intramolecular aglycon transfer reaction. , Allowed α-selective glycosylation. Examples of the protecting group that causes an intramolecular aglycone transfer include a p-methoxybenzyl group and a naphthyl group as described above.
Furthermore, the present inventors have decided to prevent the ring inversion of xylose by protecting the 3rd and 4th positions of xylose with hydroxyl groups such as diacetylate using 2,3-butandion and ring-fixing them.
Through these measures, the present inventors have solved the above problems and succeeded in stereoselectively synthesizing the α-glycoside represented by the formula (4).

The disaccharide derivative of the present invention represented by the formula (1) can be obtained by appropriately protecting the hydroxyl groups of the disaccharide of the formula (4) obtained by the above method. The selection and attachment / detachment of protecting groups is within the skill of those skilled in the art.

Although the disaccharide products can be obtained stereoselectively by the methods described in paragraphs 0014 to 0025 above, there is a problem that the yield is low. Therefore, the present inventors combine a xylose derivative (sugar donor) represented by the formula (2') and a glucose derivative or a glucuronic acid derivative (sugar acceptor) represented by the formula (3') in a mixed acetal (intramolecular). A mixture containing α-glycoside and β-glycoside represented by the formula (4') was obtained by direct condensation without a method via aglycon transfer). Isolation of the desired α-glycoside was difficult. However, as a result of diligent studies on the solubility of the desired α-glycoside-containing diastereomeric mixture obtained by removing the protecting group by hydrolysis with trifluoroacetic acid (TFA) shown in Scheme 2 below, the desired α was obtained. -While glycosides are soluble in methanol but insoluble in chloroform, the by-product β-glycosids, on the contrary, have led to the discovery of the fact that they are soluble in chloroform. Utilizing this property, we succeeded in obtaining only the desired α-glycoside in a higher yield than the methods described in paragraphs 0014 to 0025 above.

Therefore, the present invention, in a further aspect,
Equation (2'):

Wherein, X 2 ^'is a leaving group, Y' is a group for fixing the conformation of xylose residues, R 6 ^'is a protecting group for a hydroxyl group]
With the xylose derivative represented by
Equation (3'):

Wherein, X 3 ^'is a leaving group or a protected hydroxyl group, R ^7' is a protecting group for a hydroxyl group, R 8 ^'is a protecting group for a hydroxyl group, Y ^4' represents a carboxyl group or hydroxymethyl A group that is a precursor of a group]
From the glucose derivative or glucuronic acid derivative indicated by
Equation (4'):

^{^{Wherein, X 3 ', R 7'}} , R 8 ' and ^{Y 4'} are the same as defined above
A method for producing an α-glycoside represented by the following steps 1) to 3) :.
Step 1) The xylose derivative represented by the formula (2') and the glucose derivative or the glucuronic acid derivative represented by the formula (3') are directly condensed to obtain a mixture of α1 → 3 glycoside and β1 → 3 glycoside. ,
Step 2) Deprotect the mixture obtained in Step 1) and then
Step 3) Provided is a method comprising separating α-glycoside from the mixture deprotected in step 2).

The sugar donor 103 used for condensation gives the products 208a and 208b in the same yield in any configuration of αβ (Method 1 and Method 2 in Scheme 2 below). Therefore, the stereochemistry of the sugar donor 103 used is not limited and may be a mixture thereof (Method 3 of Scheme 2 below).

'In the sugar donor represented by, X 2 Equation ^(2)' and Y 'are respectively the same as X ² and Y have been described with respect to Formula (2). In formula (2'), the two oxygen atoms displayed on the left side are included in the Y'group. R 6 ^'is a protecting group for a hydroxyl group. R 6 ^'is more preferably adjacent groups is a protecting group with no involvement ability hydroxyl, acyl group which neighboring group participation ability may occur exclusively β- glycoside. Especially Examples of suitable R ^{6 'is} benzyl, p- methoxybenzyl group, p- methoxyphenyl group, p- nitrobenzyl group, an allyl group, a methoxymethyl group, tert- butyldimethylsilyl group, triisopropylsilyl Examples include, but are not limited to these. Preferred R ^{6 'include} p- methoxybenzyl group, but is not limited thereto.
In the present disclosure, a hydroxyl-protecting group capable of involving an adjacent group (Kyoritsu Publishing, Chemistry Encyclopedia) is a hydroxyl-protecting group generalized as -C (= O) R, as understood by those skilled in the art. According to the example of the formula (2'), the oxygen atom attacks and forms a carbocation generated on the 1-position carbon atom after the 1-position ^{leaving group (X 2') is eliminated.} The ring structure to be formed covers the same plane (cis plane) as the hydroxyl group at the 2-position, so that it is a protecting group having a function of exclusively attacking the receptor from the opposite side (trans side). Protecting groups that do not fall under this description are interpreted as protecting groups that are not capable of involving adjacent groups.

'In sugar acceptor represented by, R 7 Equation ^(3)', but is a protecting group for a hydroxyl group, in order to form the β- glycoside in the condensation between sugar, R 7 ^'is a neighboring group participation ability It is preferably a hydroxyl-protecting group. As a protective group for a hydroxyl group capable of involving an adjacent group, a group forming an acyl group as -C (= O) R, for example, an acetyl group in which R is an alkyl or an aryl, is substituted with 1 to 3 halogen atoms. Acetyl group, benzoyl group, benzoyl group substituted with 1-5 halogen, nitro group and / or alkyl group with 1-3 carbon atoms, and lebrinoyl group, and benzyloxycarbonyl group, allyloxycarbonyl group, 2 , 2,2-Trichloroethoxycarbonyl group and the like, but are not limited thereto. Preferred R ^7's are acetyl groups, acetyl groups substituted with 1 to 3 halogen atoms, benzoyl groups, 1 to 5 halogens, nitro groups and / or alkyl groups with 1 to 3 carbon atoms. Examples include, but are not limited to, benzoyl groups. X ³ ', ^{R 8'} and ^{Y 4} 'are respectively the same as ^X 3, ^{R 8} and ^{Y 4} which has been described with respect to equation (3). For example, X 3 ^'hydroxyl group is protected with p- methoxyphenyl, R ^7' and R ^{8 'is} acetyl group, Y ^4' may be a carboxylate group.

The condensation reaction in step 1) can be carried out using a known condensing agent (glycosylation promoter). Examples of known condensing agents include NIS-AgOTf, NIS-TfOH, MeOTf, CuBr ₂ -AgOTf-nBu ₄ NI, NBS-AgOTf and the like, but NIS-AgOTf is preferable. In the presence of these condensing agents is a leaving group X ^{2 'activates.} Direct condensation means that a substituent of a sugar donor and a substituent of a sugar acceptor form a bond without interposing another compound or group. In particular, in the present disclosure, direct condensation refers to condensation that forms a 1 → 3 glycosidic bond from the beginning, as opposed to an embodiment mediated by an intracellular aglycone transfer.

Deprotection in step 2) can be performed by a known method. Preferably, the 2-, 3-, and 4-positions of xylose residues are deprotected to triol. Among such triols, α-glycoside is a disaccharide derivative of the formula (4'). Moreover, the disaccharide derivative of the formula (4') can be easily separated by using a solvent described later. Acid is used as the drug used for such deprotection. As such an acid, trifluoroacetic acid (TFA), trichloroacetic acid and the like are preferable, and TFA is particularly preferable.

In step 3), α-glycoside is separated from the deprotected glycoside mixture (diastereomeric mixture of deprotected triol) obtained in step 2). As a method for separating diastereomers (α-glycoside and β-glycoside), a method using chromatography, a solvent extraction method, or the like can be used. If the separated α-glycoside is different from the disaccharide derivative of the formula (4'), it may be protected and / or deprotected by a known method to obtain the disaccharide derivative of the formula (4'). Good. In particular, the α-glycoside (disaccharide derivative of formula (4')) obtained as a triol by deprotecting the 2-, 3-, and 4-positions of xylose residues is insoluble in chloroform but in methanol. It is soluble and β-glycoside is soluble in chloroform (relatively low solubility in methanol). Taking advantage of this property, the desired α-glycoside can be separated by treating the diastereomeric mixture with chloroform to remove the dissolved β-glycoside. Alternatively, the desired α-glycoside may be separated by treating the diastereomer mixture with methanol to recover the dissolved α-glycoside.

A specific example of the above-mentioned new method for producing α-glycoside is shown in Scheme 2 below.

[In the above scheme, MBn is -CH ₂ C ₆ H ₄ OCH ₃ (p), MP is -C ₆ H ₄ OCH ₃ (p), Me is -CH ₃ , and Ac is -C. (= O) CH ₃ , NIS is N-iodosuccinate imide, AgOTf is silver trifluoromethanesulfonate, TFA is trifluoroacetic acid]

In a further embodiment, the present invention directly condenses the xylose derivative represented by the above formula (2') with the glucose derivative or glucuronic acid derivative represented by the above formula (3') to form an α-glycoside and a β-glycoside. Disaccharides (xylose α1-3 glucuronic acid, in the disaccharides, xylose residues and glucurons) comprising obtaining a mixture of, optionally deprotecting the mixture, and then separating the α-glycoside from the mixture. The acid residue may be derivatized).
Deprotection, if desired, means removing some (at least one) or all of the protecting groups. Derivatization of sugars is known to those of skill in the art. For example, the disaccharide may contain a known protecting group or may contain a known leaving group. Examples of the disaccharide protecting group and leaving group include those described above, but are not limited thereto.

2. Disaccharide Derivative Oligomer The disaccharide derivative of the present invention is a structural unit for the synthesis of postphosphate sugar chains on dystroglycan that anchors muscle cells and basement membrane via laminin (referred to as XG unit in the present specification). Can be used as). By oligomerizing the disaccharide derivative, the postphosphate sugar chain can be reconstructed. The disaccharide derivative of the present invention is an essential synthetic unit for reconstructing a postphosphate sugar chain.

A tetrasaccharide derivative can be obtained by condensing disaccharide derivatives with each other. A hexasaccharide derivative can be obtained by condensing a tetrasaccharide derivative and a disaccharide derivative and, if desired, deprotecting the tetrasaccharide derivative. The hexasaccharide derivative and the disaccharide derivative can be condensed and deprotected if desired to obtain a octasaccharide derivative. Alternatively, the tetrasaccharide derivatives can be condensed with each other and deprotected if desired to obtain a tetrasaccharide derivative. In this way, the oligomer of the disaccharide derivative can be synthesized.

Therefore, in a further aspect, the present invention comprises using the disaccharide derivative represented by the formula (1) as an intermediate (XG unit).

[In the formula, X ⁴ is a leaving group or a protected hydroxyl group, and n is an integer greater than or equal to 1.]
Provided is a method for producing an oligomer of the XG unit represented by. It will be understood by those skilled in the art that the upper limit of n is not particularly limited because the oligomeric linking reaction can be simply repeated. n can be, for example, 300 or less, 200 or less, 100 or less, 50 or less, 20 or less, and the like.

^{When X 4 of the} compound represented by the formula (5) is a leaving group, X ⁴ is a leaving group similar ^{to X 1 and the like described above.} When X ⁴ is a protected hydroxyl group, X ⁴ may contain the same protecting groups as ^{R 1} and the like described above.

Specifically, the disaccharide derivative represented by the formula (1) is used as a sugar donor, and the formula (1'):

[In the formula, Y ⁵ is a group that is a precursor of a carboxyl group or a hydroxymethyl group, X ⁵ is a leaving group or a protected hydroxyl group, and R ⁹ to R ¹² are a protecting group of a hydroxyl group].
A tetrasaccharide derivative can be obtained by using the disaccharide derivative represented by (1) as a sugar receptor and condensing them. By the same method, the tetrasaccharide derivatives are condensed with each other, or the disaccharide derivative represented by the formula (1) and the tetrasaccharide derivative are condensed, or the tetrasaccharide derivative and the disaccharide derivative represented by the formula (1') are condensed. Or by repeating the condensation of these oligomers with the oligomer and / or the disaccharide derivative, an oligomer derivative having a longer chain length can be obtained. These oligomer derivatives can be deprotected by a known method to obtain an oligomer represented by the formula (5) having a longer chain length.

In equation (1'), Y ⁵ is a group similar to Y ^{1 etc. described above.} Examples of Y ⁵ include, but are not limited to, ^{COOZ 3} or CH ₂ OZ ^4. Here, Z ³ is an alkyl group, preferably an alkyl group having 1 to 3 carbon atoms. More preferred Z ³ groups are methyl. Z ⁴ is a normal hydroxyl group protecting group. X ⁵ is a leaving group or a protected hydroxyl group. R ⁹ to R ¹² are hydroxyl-protecting groups, which are the same groups as ^{R 1 and the like described above.} Preferred X ^5, trichloroacetimidate benzyloxy, alkylthio, arylthio, halogen, such as pentenyloxy, although leaving groups include, but are not limited to commonly used in sugar chain synthesis. Preferred R ⁹ to R ¹² are generally used for the synthesis of sugar chains such as benzyl containing a substituted benzyl, allyl, lebrinoyl, benzoyl containing a substituted benzoyl, acetyl containing a substituted acetyl, allyloxycarbonyl, and trialkylsilyl. Protecting groups include, but are not limited to. Preferred R ^9, benzoyl including substituted benzoyl with neighboring group participation ability, acetyl including substituted acetyl, although such levulinoyl include, but are not limited to. Additional examples of protecting groups which neighboring group participation ability has been described for R ^{7 'groups} to the aforementioned, they can be used in R ⁹ groups.

A specific example of a procedure for synthesizing a tetrasaccharide derivative by condensing the disaccharide derivative represented by the formula (1) as a sugar donor and the disaccharide derivative represented by the formula (1') as a sugar receptor is described below. Is shown in Scheme 3. The oligomer 206b obtained in the specific example below can be deprotected by a known method to obtain the oligomer (n = 1) represented by the formula (5).

[In the above scheme, Lev is -C (= O) C ₂ H ₄ C (= O) CH ₃ , Bz is -C (= O) C ₆ H ₅ , and MP is -C ₆ H ₄ OCH ₃ (p), Me is -CH ₃ , MBz is -C (= O) C ₆ H ₄ CH ₃ (p)]

The oligomer 206b obtained in the above specific example is deprotected by treating it with a known method, for example, a base such as an aqueous sodium hydroxide solution to obtain the oligomer (n = 1) represented by the formula (5). Can be done.

In a further aspect of the present invention, the disaccharide derivative represented by the above formula (1) is used as a sugar donor, the disaccharide derivative represented by the above formula (1') is reacted as a sugar acceptor, and then, if desired, the disaccharide derivative is reacted. Provided is a method for producing a tetrasaccharide represented by the formula (5), which may be derivatized, which comprises deprotecting the obtained tetrasaccharide derivative.
Deprotection, if desired, means removing some (at least one) or all of the protecting groups. Derivatization of sugars is known to those of skill in the art. For example, the tetrasaccharide may contain a known protecting group or may contain a known leaving group. Examples of the tetrasaccharide protecting group and leaving group include those described above, but are not limited thereto.

3. 3. Synthesis of a tetrasaccharide that binds a xylose-glucuronic acid repeating structure of a postphosphate sugar chain to a sugar chain extended from α-dystroglycan The present invention has a further embodiment of the formula (9):

Provided are tetrasaccharides or derivatives thereof. The tetrasaccharide of the formula (9) is a portion that binds the xylose-glucuronic acid repeating structure of the postolinate sugar chain to the sugar chain extended from α-dystroglycan. The postphosphate sugar chain can be reconstructed using the above disaccharide derivative, its oligomer, and the tetrasaccharide of the formula (9). The reconstructed sugar chain can be used as a medicine for the treatment and prevention of sugar chain abnormal muscular dystrophy. Examples of the derivative of the formula (9) include a compound in which at least a part or all of the hydroxyl groups in the formula (9) are protected by a protecting group, and a carboxyl group in the formula (9) is alkyl esterified. Examples include, but are not limited to, compounds. Specific examples of the tetrasaccharide derivative of the formula (9) include, but are not limited to, compounds 404 and 405 in the scheme below.

In yet another aspect of the present invention, the formula (7):

Provided is a method for producing the tetrasaccharide shown by.

Compound can be used and known as a leaving group X ¹ in (7), trichloroacetimidate benzyloxy, alkylthio, arylthio, halogen, a leaving group is exemplified commonly used in the sugar chain synthesis such pentenyloxy However, it is not limited to these. Preferred leaving groups X ¹ include, but are not limited to, those that form thioglycosides such as trichloroacetimideyloxy, alkylthio and phenylthio. As the compound (7) protecting group X ^{2 ~} X ⁶ hydroxyl groups in benzyl including substituted benzyl, allyl, levulinoyl, benzoyl, including substituted benzoyl, acetyl including substituted acetyl, allyloxycarbonyl, sugar chain such as trialkylsilyl Protecting groups commonly used in synthesis include, but are not limited to. The alkyl group in compound (7) is preferably an alkyl group having 1 to 3 carbon atoms, and is typically a methyl group. In the specific example of ¹ , X 1 of compound (7) is trichloroacetimideyloxy, X ² to X ⁶ are acetyl, and X ⁷ is methyl.

As the compound (8) of the hydroxyl groups in the protecting group X ^{8 ~} X ^13, 2,3-position hydroxyl group of xylose is protected, as long as the protecting group of ribitol side is not no or changed is removed, commonly used in the sugar chain synthesis Protecting groups can be used. Examples of such protecting groups include, but are not limited to, benzyl containing substituted benzyl, allyl, lebrinoyl, benzoyl containing substituted benzoyl, acetyl containing substituted acetyl, allyloxycarbonyl, trialkylsilyl and the like. The hydroxyl groups at the 2- and 3-positions of compound (8) may be protected by acetals. In one embodiment, compound (8) X ⁸ is protected by allyl, X ⁹ and X ¹⁰ are protected by benzyl, X ¹¹ is protected by TBDPS, and the hydroxyl groups at the 2- and 3-positions are protected by isopropyrine acetals (ie,). X ¹² and X ¹³ together form an isopropyrine acetal).

The compound of formula (7) may be derived from the disaccharide derivative of formula (1) using a known method.

The compound of formula (8) can be prepared using a known method, for example, of formula (10):

[In the formula, Lev is -C (= O) C ₂ H ₄ C (= O) CH ₃ , Bn is -CH ₂ C ₆ H ₅ , All is -CH ₂ CH = CH ₂ . TBDPS is -Si (C ₆ H ₅ ) ₂ tert-C ₄ H ₉ ]
It may be derived from the disaccharide derivative represented by.

A specific example of the above method is shown in Scheme 4 below.

[In the above scheme, Lev is -C (= O) C ₂ H ₄ C (= O) CH ₃ , Et is -C ₂ H ₅ , and Ac is -C (= O) CH ₃ . , MBz is -C (= O) C ₆ H ₄ CH ₃ (p), All is -CH ₂ CH = CH ₂ , CSA is camphor sulfonic acid, DMF is dimethylformamide, and THF is -Si (C ₆ H ₅ ) ₂ tert-C ₄ H ₉ , THF is tetrahydrofuran, TMSOTf is trimethylsilyl trifluoromethanesulfonate, All is -CH ₂ CH = CH ₂ , TBAF is tetrabutyl. Ammonium fluoride]

The compound represented by the formula (7) is obtained by introducing a leaving group such as _{-OC (NH) CCl 3} into the 1-position of glucuronic acid of the compound having the protecting group of the hydroxyl group of the disaccharide derivative of the formula (1) as an acetyl group. You may get it. The compound represented by the formula (10) may be derived from xylose. The lebrinoyl protecting groups at the 2-, 3- and 4-positions of the compound represented by the formula (10) are deprotected, and then a ring is formed between the hydroxyl groups at the 2- and 3-positions and 2-methoxypropene. Obtain the compound shown in (8). The compound (sugar donor) represented by the formula (7) is reacted with the compound (sugar acceptor) represented by the formula (8), and then deprotection is performed by a known method to obtain the tetrasaccharide represented by the formula (9). obtain.

In a further aspect of the present invention, the disaccharide donor represented by the above formula (7) is reacted with the disaccharide receptor represented by the above formula (8), and then, if desired, the obtained tetrasaccharide derivative is obtained. Provided is a method for producing a tetrasaccharide represented by the formula (9), which may be derivatized, including deprotection.
Deprotection, if desired, means removing some (at least one) or all of the protecting groups. Derivatization of sugars is known to those of skill in the art. For example, the tetrasaccharide may contain a known protecting group or may contain a known leaving group. Examples of the tetrasaccharide protecting group and leaving group include those described above, but are not limited thereto.

The reaction conditions shown in the above description and scheme are examples, and those skilled in the art can appropriately change these reaction conditions.

Unless otherwise specified, the meanings of the terms in this specification are the same as those commonly understood in fields such as biochemistry, biology, chemistry, pharmacy, and medicine.

The present invention will be described in more detail and concretely by showing examples below, but the examples are merely exemplary explanations and do not limit the scope of the present invention.

Example 1 Synthesis of disaccharide derivative (1) Synthesis of Dodecyl 3,4-O- (2,3-dimethoxybutan-2,3-diyl) -1-thio-α-D-xylopyranoside (102a) MeOH (94 mL) To a known compound (101a, 6.19 g, 18.5 mmol) dissolved in, trimethyl orthoformate (18 mL), 2,3-butandione (6.5 mL) and a catalytic amount of camphorsulfonic acid were added, and the mixture was stirred at 50 ° C. overnight. did. Triethylamine was added and concentrated, and the concentrated residue was purified by silica gel column chromatography (toluene: ethyl acetate = 20: 1 to 5: 1) to obtain 102a (2.97 g, 27%).

(X) Synthesis of Dodecyl 3,4-O- (2,3-dimethoxybutan-2,3-diyl) -1-thio-β-D-xylopyranoside (102b) Known compound dissolved in MeOH (30 mL) (101b, To 2.10 g, 6.28 mmol) was added trimethyl orthoformate (6 mL), 2,3-butandione (2.4 mL) and a catalytic amount of camphorsulfonic acid, and the mixture was stirred at 40 ° C. overnight. Triethylamine was added and concentrated, and the concentrated residue was purified by silica gel column chromatography (toluene: ethyl acetate = 20: 1 to 3: 1) to obtain 102b (2.52 g, 89%).

(2) Synthesis of 3,4-O- (2,3-dimethoxybutan-2,3-diyl) -2-O- (4-methoxy) benzyl-1-thio-α-D-xylopyranoside (103a) 102a, 941.9 mg, 2.099 mmol) was dissolved in dimethylformamide (11 mL), sodium hydride (55%, 106.3 mg, 2.44 mmol) was added, and the mixture was stirred for 2 hours. To this, 4-methoxybenzyl chloride (0.57 mL, 5.6 mmol) was added, and the mixture was stirred for 4 hours. Ice and 1M aqueous ammonium chloride solution were added to the reaction mixture, and the mixture was extracted with ethyl acetate. The concentrated residue was purified by silica gel column chromatography (n-hexane: ethyl acetate = 50: 1 to 10: 1) to obtain 103a (743.0 mg, 62%).

(2) Synthetic compound of 3,4-O- (2,3-dimethoxybutan-2,3-diyl) -2-O- (4-methoxy) benzyl-1-thio-β-D-xylopyranoside (103b) ( 102b, 2.52 g, 5.62 mmol) was dissolved in dimethylformamide (28 mL), sodium hydride (55%, 0.50 g, 11.5 mmol) was added, and the mixture was stirred for 2 hours. To this, 4-methoxybenzyl chloride (1.6 mL, 8.9 mmol) was added, and the mixture was stirred for 3 hours. Ice and 1M aqueous ammonium chloride solution were added to the reaction mixture, and the mixture was extracted with ethyl acetate. The concentrated residue was purified by silica gel column chromatography (n-hexane: ethyl acetate = 8: 1 to 6: 1) to obtain 103b (2.43 g, 76%).

(3) Synthesis of Methyl (4-methoxyphenyl 2,4-di-O-acetyl-β-D-glucopyranoside) uronate (113a) Known compound (111, 22.75 g, 51.66 mmol) dissolved in MeOH (120 mL) 1.0 M NaOH (120 mL) was added to the mixture, and the mixture was stirred overnight at room temperature. Then, it was neutralized with 1.0 M HCl, concentrated and dried. _{Ac 2} O (500 mL) and I ₂ (0.24 g) were added to the obtained dried product, and the mixture was stirred at room temperature for 8 hours. An appropriate amount of MeOH, ice and 1M hypo (sodium thiosulfate) were added, the mixture was extracted with ethyl acetate, and dried over _{anhydrous plate 4.} The insoluble material was filtered and concentrated, and purified by silica gel column chromatography (toluene: ethyl acetate = 6: 1) to obtain a product. The obtained product was dissolved in MeOH 400 mL and heated to reflux for one week. The reaction mixture was concentrated and purified by silica gel column chromatography (toluene: ethyl acetate = 4: 1) to obtain 3.56 g (3 step yield 17%) of 113a.

(4) Synthesis of Methyl (4-methoxyphenyl 2,4-di-O-benzoyl-β-D-glucopyranoside) uronate (113b) A known compound (111, 11) dissolved in a mixed solvent of THF (212 mL) and water (31 mL). 1.25 M LiOH (296 mL) was added to 23.92 g (54.31 mmol) at 0 ° C., and the mixture was stirred for 6 hours. Then, it was neutralized with 1.0 M HCl, concentrated and dried. The obtained dried product was dissolved in DMF (550 mL), benzoic anhydride (172.24 g, 761.34 mmol) was added, and the mixture was stirred at 79 ° C. for 3 hours. Then, pyridine (226 mL) and DMAP (3.40 g, 27.8 mmol) were added, and the mixture was stirred overnight at room temperature. Adding an appropriate amount of ice, and extracted with ethyl acetate, washed with saturated brine and ice-cold 1M HCl, and the organic layer was dried over anhydrous MgSO _4. The insoluble material was filtered and concentrated, the product was dissolved in MeOH (563 mL), sodium acetate (7.86 g, 95.8 mmol) was added, and the mixture was heated and stirred for 5 hours. The reaction mixture was concentrated, extracted with chloroform, post-treated by a conventional method, and then the residue was purified by silica gel column chromatography (n-hexane: ethyl acetate = 8: 1 to 0: 1) to obtain 113b at 10. 9.9 g (3 step yield 38%) was obtained.

(5) Methyl (2,3,4-tri-O-acetyl-α-D-xylopyranosyl)-(1 → 3) -β- (4-methoxyphenyl2,4-di-O-acetyl-β-D-glucopyranosid) ) Synthesis of uronate (204a)
_{Molecular Sieves 4A (1.5 g) was added to a CH 2} Cl ₂ (5 mL) solution of DDQ (477.2 mg, 2.102 _{mmol), and a CH 2} Cl ₂ (10 mL) solution of 113a (535.7 mg, 1.354 mmol) was added. Was added, and the mixture was stirred at room temperature for 1 hour. A solution of 103b (967.4 mg, 1.700 mmol) of CH ₂ Cl ₂ (7 mL) was added thereto at 0 ° C. with stirring, and then the reaction was continued at room temperature for 4 hours. A 0.1 M aqueous ascorbic acid solution was added to the reaction solution, and the filtrate filtered through diatomaceous earth was extracted with chloroform. The organic layer is aqueous 0.1M ascorbic acid, washed with a saturated aqueous sodium bicarbonate solution and saturated brine, and dried over anhydrous MgSO _4. The insoluble material was filtered off, and the concentrated residue of the filtrate was purified by silica gel column chromatography (n-hexane: ethyl acetate = 6: 1 to 1:10) to obtain 1.074 g (83%) of mixed acetal (201a). It was. NIS (525.5 mg, 2.34 mmol) and AgOTf (195.3 mg, 760 μmol) were _{suspended in CH 2} Cl ₂ (22 mL), and stirred in the presence of molecular sieves 4A (2.3 g) for 1 hour in the dark. .. _{A CH 2} Cl ₂ (220 mL) solution of the mixed acetal (201a, 1.074 g) was added dropwise at −20 ° C. and stirring was continued for 2 hours. An appropriate amount of 1M hypo, saturated aqueous sodium hydrogen carbonate and saturated brine were added to the reaction mixture, and the filtrate filtered through diatomaceous earth was extracted with chloroform. The organic layer was washed with 1M hypo, saturated aqueous sodium hydrogen carbonate and saturated brine, and dried over anhydrous plate ₄ . The insoluble material was filtered off, and the concentrated residue of the filtrate was purified by silica gel column chromatography (n-hexane: ethyl acetate = 30: 1 to 1:10 to ethyl acetate: methanol = 30: 1), and the disaccharide fraction (disaccharide fraction) 753.9 mg) was obtained. Of this, 90% trifluoroacetic acid (15 mL) was added to 299.7 mg at 0 ° C., and the mixture was stirred. The reaction solution was returned to room temperature over 2 hours, and the reagent was distilled off under reduced pressure. Pyridine (3 mL) and acetic anhydride (3 mL) were added to the concentrated residue, and the mixture was stirred for 3 hours. The reagent was distilled off under reduced pressure, and the concentrated residue was purified by silica gel column chromatography (toluene: ethyl acetate = 10: 1 to 2: 3) to obtain 85.6 mg (24%) of 204a.

(6) Synthesis of Methyl α-D-xylopyranosyl- (1 → 3) -β- (4-methoxyphenyl 2,4-di-O-benzoyl-β-D-glucopyranosid) uronate (202ba) Molecular Sieves 4A (1. _{To a suspension of CH 2} Cl ₂ (3 mL) of 74 g) and DDQ (474.2 mg, 2.087 mmol) was added a solution of 113b (706.3 mg, 1.352 mmol) CH ₂ Cl ₂ (7 mL) and 1 at room temperature. Stirred for hours. A solution of 103b (962.9 mg, 1.693 mmol) of CH ₂ Cl ₂ (8 mL) was added thereto at 0 ° C. with stirring, and then the reaction was continued overnight at room temperature. The reaction mixture was cooled to 0 ° C. again, DDQ (160.4 mg, 706 μmol) was added, and the mixture was stirred for 5 hours. After returning to room temperature and performing the same post-treatment as the synthesis of 204a, the concentrated residue was subjected to gel filtration column chromatography (chloroform: methanol = 1: 1) and silica gel column chromatography (toluene: ethyl acetate = 1: 0 to 2: 1). To obtain 435.5 mg (yield 30%) of mixed acetal (201b). NIS (124.6 mg, 553.8 μmol) and AgOTf (61.6 mg, 0.24 mmol) were _{suspended in CH 2} Cl ₂ (4 mL) in the presence of molecular sieves 4A (0.43 g) in the presence of light-shielded mixed acetal. _{A solution of CH 2} Cl ₂ (40 mL) (201b, 232.9 mg) was added dropwise at −20 ° C., and stirring was continued for 3 hours. The same post-treatment as the synthesis of 204a was carried out and purified by gel filtration column chromatography (chloroform: methanol = 1: 1) to obtain a product (156.7 mg). 90% trifluoroacetic acid (8 mL) was added thereto at 0 ° C., and the mixture was stirred for 2.5 hours. The reagent was evaporated to room temperature under reduced pressure and purified by gel filtration column chromatography (chloroform: methanol = 1: 1) to obtain 202ba (49.4 mg, 75.5 μmol, 2-step yield 35%).

(7) Methyl [2,4-di-O- (4-methyl) benzoyl-α-D-xylopyranosyl]-(1-3) -β- (4-methoxyphenyl 2,4-di-O-benzoyl-β -D-glucopyranosid) uronate (203b1), Methyl [3,4-di-O-(4-methyl) benzoyl-α-D-xylopyranosyl]-(1-3) -β- (4-methoxyphenyl 2,4- Synthesis of di-O-benzoyl-β-D-glucopyranosid) uronate (203b2) Compound 202ba (239.2 mg, 365.4 μmol) was dissolved in toluene (13 mL) to dissolve dibutyltin oxide (IV) (327.6 mg, 1). .32 mmol) was added and reacted in a Dean Stark apparatus for 2.5 hours. The reaction mixture was ice-cooled, 4-methylbenzoyl chloride (122 μL, 923 μmol) was added, and the mixture was stirred overnight at room temperature. Saturated aqueous sodium hydrogen carbonate was added to the reaction mixture, the mixture was extracted with ethyl acetate, and post-treated by a conventional method. The concentrated residue was purified by gel filtration column chromatography (chloroform: methanol = 1: 1), the chloroform solution of the product was washed with 1M HCl and saturated brine, and the concentrated residue (381.9 mg) of the organic layer was added to toluene (381.9 mg). It was dissolved in 12.5 mL) and methanol (2.5 mL). This solution was ice-cooled, 2M TMS diazomethane (660 μL, 11.6 μmol) was added, and the mixture was concentrated under reduced pressure after 1 hour. The residue was purified by gel filtration column chromatography (chloroform: methanol = 1: 1) and silica gel column chromatography (toluene: ethyl acetate = 10: 0-0: 1), and 203b1 (202.9 mg) and 203b2 (42. 6 mg,) were obtained in yields of 62% and 25%, respectively.
203b1: ESI-HRMS m / z [(M + Na) ⁺ ]: calcd. For C ₄₉ H ₄₆ _{NaO 16} : 913.2678; found, 913.2678.
203b2: ESI-HRMS m / z [(M + Na) ⁺ ]: calcd. For C ₄₉ H ₄₆ _{NaO 16} : 913.2678; found, 913.2658.

(8) Methyl [3-O-levulynoyl-2,4-di-O- (4-methyl) benzoyl-α-D-xylopyranosyl]-(1 → 3) -β- (4-methoxyphenyl 2,4-di Synthesis of -O-benzoyl-β-D-glucopyranosid) uronate (204b) Compound (203b 1,111.4 mg, 125 μmol) was dissolved in pyridine (640 μL) and 1 and 2 of 1 M levulinic acid (325 μl, 0.313 mmol). -A small amount of dichloroethane solution and DMAP were added, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was neutralized with saturated brine, _{extracted with CHCl 3} and post-treated by a conventional method. The obtained residue was purified by silica gel column chromatography (n-hexane: ethyl acetate = 50: 1 to 0: 1) to quantitatively obtain 204b (124.8 mg).

(9) Methyl (2,3,4-tri-O-acetyl-α-D-xylopyranosyl)-(1 → 3) -β- (4-methoxyphenyl 2,4-di-O-acetyl-β-D- Glucopyranosylimidate) compound of uronate (205a) (204a, 83.4mg , a 127Myumol) was dissolved in a mixed solution of acetonitrile (4 mL) and _H 2 O (1mL), there the CAN (203.6mg, 371.4μmol) In addition, the mixture was stirred at room temperature for 1.5 hours. 0.1 M ascorbic acid was added to the reaction mixture, CHCl ₃ extraction and washing with saturated brine were carried out, and post-treatment was carried out by a conventional method. The obtained residue was purified by silica gel column chromatography (toluene: ethyl acetate = 5: 1 to 2: 3) to obtain a product (73.6 mg). This was dissolved in dichloromethane (2 mL), CCl ₃ CN (127 μL, 1.25 mmol) was added, and the mixture was cooled to 0 ° C. Then, 2 drops of DBU were added, the temperature was returned to room temperature, and the mixture was stirred for 1 hour. The reaction mixture was purified by silica gel column chromatography (toluene: ethyl acetate = 10: 1 to 1: 1) to obtain 205a (72.8 mg) in a yield of 82% (2 steps).
¹ H-NMR δ _H (CDCl ₃ ): 8.72 (s, 1H, NH), 6.67 (d, 1H, J _1,2 = 3.54 Hz, GlcA-1), 5.35 (brt, 1H, J = 10.08 Hz, Xyl-3), 5.33 (d, 1H, J _1,2 = 3.72 Hz, Xyl-1), 5.30 (dd, 1H, J _3,4 = 9.37 Hz, J _4,5 = 10.20 Hz, GlcA-4) , 5.10 (dd, 1H, J _2,3 = 9.90 Hz, GlcA-2), 4.95 (ddd, 1H, J _3,4 = 10.14 Hz, J _4,5a = 6.42 Hz, J _4,5e = 9.60 Hz, Xyl-4), 4.72 (dd, 1H, J _2,3 = 10.26 Hz, Xyl-2), 4.35 (brt, 1H, J = 9.63 Hz, GlcA-3), 4.35 (d, 1H, GlcA-5) , 3.76 (m, 2H, Xyl-5a, e), 3.73 (s, 3H, COOMe), 2.09, 2.04, 2.04, 2.02, 2.01 (each s, 3Hx5, 5Ac)

(10) Methyl [3-O-levulynoyl-2,4-di-O- (4-methyl) benzoyl-α-D-xylopyranosyl]-(1 → 3) -β- (4-methoxyphenyl 2,4-di -O-benzoyl-β-D- glucopyranosylimidate) compound of uronate (205b) (204b, 124.8mg , dissolved 126.2Myumol) in a mixed solution of acetonitrile (5.8 mL) and _H 2 O (1.4mL) Then, CAN (362.2 mg, 660.7 μmol) was added thereto, and the mixture was stirred at room temperature for 4.5 hours. 0.1 M ascorbic acid was added to the reaction mixture, CHCl ₃ extraction and washing with saturated brine were carried out, and post-treatment was carried out by a conventional method. The obtained residue was purified by silica gel column chromatography (n-hexane: ethyl acetate = 50: 1 to 0: 1) to obtain a product (110.8 mg). This was dissolved in dichloromethane (1 mL), CCl ₃ CN (125 μL, 1.25 mmol) was added, and the mixture was cooled to 0 ° C. Then, 1 drop of DBU was added, the temperature was returned to room temperature, and the mixture was stirred for 1 hour. The reaction mixture was purified by silica gel column chromatography (toluene: ethyl acetate = 50: 1 to 0: 1) to obtain 205b (109.7 mg) in a yield of 80% (2 steps).
¹ H-NMR δ _H (CDCl ₃ ): 8.70 (s, 1H, NH), 7.97-7.95 (m, 2H, Ar), 7.64-7.58 (m, 4H, Ar), 7.54-7.51 (m, 3H, Ar) Ar), 7.33-7.29 (m, 2H, Ar), 7.22 (brt, 1H, J = 7.80 Hz, Ar), 7.16 (d, 1H, J = 7.98 Hz, Ar), 7.11 (d, 1H, J = 7.98 Hz, Ar), 6.84 (d, 1H, J _1,2 = 3.78 Hz, GlcA-1), 5.60 (brt, 1H, J = 9.90 Hz, Xyl-3), 5.59-5.56 (m, 2H, GlcA) -2, 4), 5.50 (d, 1H, J _1,2 = 3.90 Hz, Xyl-1), 5.00 (dd, 1H, J _2,3 = 10.44 Hz, Xyl-2), 4.97 (m, 1H, Xyl-4), 4.72 (t, 1H, J _2,3 = J _3,4 = 9.46 Hz, GlcA-3), 4.52 (d, 1H, J _4,5 = 10.20 Hz, GlcA-5), 3.70 ( brt, 1H, J = 10.92 Hz, Xyl-5a), 3,63 (dd, 1H, J _4,5e = 6.18 Hz, J _{5a, 5e} = 11.22 Hz, Xyl-5e), 3.52 (s, 3H, COOMe) ), 2.40 (s, 6H, 2Ph Me ), 2.39-2.16 (m, 4H, 2CH ₂ ), 1.86 (s, 3H, Lev).

(X) Methyl 3,4-O- (2,3-dimethoxybutan-2,3-diyl) -2-O- (4-methoxy) benzyl-α-D-xylopyranosyl- (1 → 3) -β-( Synthesis of 4-methoxyphenyl 2,4-di-O-acetyl-α and β-D-glucopyranosid) uronate (208a, 208b)
(Condensation using α-thioglycoside: Method 1)
103a (397.8 mg, 0.699 mmol) and 113a (183.5 mg, 0.461 mmol) in a mixed solvent of toluene (3.0 mL) and dioxane (3.0 mL) of molecular sieve 4A (0.64 g). In the presence, the mixture was stirred at room temperature for 1 hour. NIS (238.2 mg, 1.059 mmol) and AgOTf (71.4 mg, 0.278 mmol) were added ^{thereto at −20 o C.} After 1.5 hours, 1M hypo, saturated _{NaCl 3} and saturated NaCl were added to terminate the reaction, the filtrate filtered through diatomaceous earth _{was extracted with CHCl 3} , and the organic layer was washed _{with saturated NaCl 3} and saturated NaCl, and anhydrous silyl ₄ It was dried in. The insoluble material is filtered off, and the concentrated residue of the filtrate is purified by gel filtration column column chromatography (chloroform: methanol = 1: 1) to obtain a mixture of 208a and 208b (245.9 mg, α58%, β12%). It was.

(Condensation using β-thioglycoside: Method 2)
103b (184.7 mg, 0.325 mmol) and 113a (86.9 mg, 0.218 mmol) in a mixed solvent of toluene (3.0 mL) and dioxane (3.0 mL) of molecular sieve 4A (0.33 g). In the presence, the mixture was stirred at room temperature for 2 hours. NIS (110.1 mg, 0.489 mmol) and AgOTf (38.4 mg, 0.150 mmol) were added thereto at −20 ° C. After 2 hours, 1M hypo, saturated _{NaCl 3} and saturated NaCl were added to terminate the reaction, the filtrate filtered through diatomaceous earth _{was extracted with CHCl 3} , and the organic layer was washed _{with saturated NaCl 3} and saturated NaCl, and dried with anhydrous _{chloride 4} . did. The insoluble material was filtered off, and the concentrated residue of the filtrate was purified by gel filtration column chromatography (chloroform: methanol = 1: 1) to obtain a mixture of 208a and 208b (120.7 mg, α60%, β12%). ..

(Condensation using α and β thioglycosides: Method 3)
About 1: 1 mixture (1.05 g, 1.85 mmol) and 113a (491.4 mg, 1.234 mmol) of 103a and 103b in a mixed solvent of toluene (10.0 mL) and dioxane (10.0 mL) is molecular. The mixture was stirred at room temperature for 1 hour in the presence of sheave 4A (1.81 g). NIS (588.3 mg, 2.615 mmol) and AgOTf (191.6 mg, 0.7458 mmol) were added thereto at −20 ° C. After 2 hours, 1M hypo, saturated _{NaCl 3} and saturated NaCl were added to stop the reaction, and the filtrate filtered through diatomaceous earth was extracted with _{CHCl 3} and dried over _{anhydrous silyl 4.} The insoluble material was filtered, concentrated and purified by gel filtration column chromatography (chloroform: methanol = 1: 1) to obtain a mixture of 208a and 208b (754.4 mg, α67%, β13%).

(X) Synthesis of Methyl α-D-xylopyranosyl- (1 → 3) -β- (4-methoxyphenyl 2,4-di-O-acetyl-β-D-glucopyranosid) uronate (202aa) Mixture of compounds 208a and 208b 90% trifluoroacetic acid (10.0 mL) was added to (754.4 mg, α: β = 67: 13), and the mixture was stirred at 0 ° C. After 2.5 hours, the mixture was concentrated and purified by silica gel column chromatography (n-hexane: ethyl acetate = 1: 1 to ethyl acetate: methanol = 10: 1) to obtain a mixture of 202aa and 202ab (474.5 mg, yield 91%). ) Was obtained. This was suspended in chloroform and filtered. The insoluble material was recovered to give 202aa (427.8 mg) (82% yield from a mixture of 208a and 208b). ¹ H-NMR δ _H (CD ₃ OD): 6.98-6.92 (m, 2H, Ar-H), 6.84-6.82 (m, 2H, Ar-H), 5.19-5.11 (m, 4H, Glc A-1) , 2,4), 4.95 (d, 1H, J _1,2 = 3.84Hz, Xyl-1), 4.27 (d, 1H, J _4,5 = 9.96Hz, Glc A-5), 4.12 (t, 1H , J _2,3 = J _3,4 = 9.24Hz, Glc A-3), 3.74,3.69 (2s, 3H × 2,2O C H ₃ ), 3.53-3.39 (m, 4H, Xyl-3,4,5ab ), 2.12, 2.07 (2s, 3H × 2,2A c).

Example 2 Synthesis of tetrasaccharide derivative (compound of formula (5) with n = 1) (1) Methyl [3-O-levulinoyl-2,4-di-O- (4-methyl) benzoyl-α-D -xylopyranosyl]-(1 → 3)-(methyl 2,4-di-O-benzoyl-β-D-glucopyranosyluronate)-(1 → 3)-[2,4-di-O-(4-methyl) benzoyl -Α-D-xylopyranosyl]-(1 → 3)-(4-methoxyphenyl 2,4-di-O-benzoyl-β-D-glucopyranosid) uronate (206b) synthesis
The disaccharide donor (205b, 109.7 mg, 106.7 μmol) and the disaccharide receptor (203b 1,90.2 mg, 101 μmol) were dissolved in dichloromethane (3.5 mL), and MSAW300 (0.31 g) was added. The mixture was stirred for 30 minutes. This was cooled to −20 ° C., TMSOTf (10 μL, 53.5 μmol) was added, and the mixture was stirred for 3 hours. Saturated aqueous sodium hydrogen carbonate was added to the reaction solution, and Celite filtration was performed. After treatment by a conventional method _{, the product was purified by gel filtration column chromatography (CHCl 3} : MeOH = 1: 1) to obtain 206b (56.3 mg, 32.1 μmol) in a yield of 32%. [α] _D + 21.0 ° (c 0.66, CHCl ₃ ). ¹ H-NMR δ _H (CDCl ₃ ): 7.92-7.899 (m, 4H, Ar), 7.66-7.62 (m, 5H, Ar), 7.59 ( d, 2H, J = 8.16 Hz, Ar), 7.55-7.50 (m, 7H, Ar), 7.41 (d, 2H, J = 7.44 Hz, Ar), 7.33-7.27 (m, 5H, Ar), 7.18 ( m, 3H, Ar), 7.15 (d, 2H, J = 8.04 Hz, Ar), 7.11 (d, 2H, J = 7.75 Hz, Ar), 7.06 (d, 2H, J = 7.92 Hz, Ar), 6.91 -6.89 (m, 2H, Ar), 6.88-6.86 (m, 2H, Ar), 6.71-6.70 (m, 2H, Ar), 5.55 (dd, 1H, J _1,2 = 7.26 Hz, J _2,3 = 8.64 Hz, H-2 ¹ ), 5.50 (brt, 1H, J = 8.94 Hz, H-4 ¹ ), 5.45 (t, 1H, J _2,3 = J _3,4 = 9.78 Hz, H-3 ⁴ ), 5.42 (brt, 1H, J = 9.45 Hz, H-4 ³ ), 5.34 (d, 1H, J _1,2 = 3.60 Hz, H-1 ² ), 5.28 (d, 1H, J _1,2 = 3.96 Hz, H-1 ⁴ ), 5.25 (brt, 1H, J = 8.52 Hz, H-2 ³ ), 5.09 (d, 1H, H-1 ¹ ), 5.00 (dd, 1H, J _1,2 = 7.98 Hz, H-1 ³ ), 4.81-4.76 (m, 3H, H-4 ² , 2 ⁴ , 4 ⁴ ), 4.71 (dd, 1H, J _2,3 = 9.60 Hz, H-2 ³ ), 4.43 ( brt, 1H, J = 9.27 Hz, H-3 ² ), 4.33 (brt, 1H, J = 8.73 Hz, H-3 ¹ ), 4.17 (brt, 1H, J = 9.08 Hz, H-3 ³ ), 4.12 (d, 1H, J _4,5 = 9.84 Hz, H-5 ³ ), 4.11 (d, 1H, J _4,5 = 9.19 Hz, H-5 ¹ ), 3.70, 3.53, 3.44 (3s) , 3Hx3, 3OMe), 3.66 (dd, 1H, J _4,5e = 5.93 Hz, J _{5a, 5e} = 11.53 Hz, H-5 ² e), 3.49 (brt, 1H, J = 10.78 Hz, H-5 ² a), 3.31 (brt, 1H, J = 11.05 Hz, H-5 ⁴ a), 3.21 (dd, 1H, J _4,5e = 6.00 Hz, J _{5a, 5e} = 11.33 Hz, H-5 ⁴ e), 2.50, 2.45, 2.39, 2.37 (4s, 12H, 4Ph Me ), 2.44-2.10 (m, 4H, 2CH ₂ ), 1.83 (s, 3H, Lev). ESI-HRMS m / z [(M + Na) ⁺ ]: calcd. for C ₉₆ H ₉₀ _{NaO 32} : 1777.5307; found, 1777.5330.

(2) α-D-Xylopyranosyl- (1 → 3) -β-D-glucopyranosyluronic acid- (1 → 3) -α-D-xylopyranosyl- (1 → 3) -4-methoxyphenyl β-D-glucopyranosyluronic acid, disodium compound of salt (207b) (206b, 15.1mg , 8.60μmol) was dissolved in a mixed solution of THF (1.4 ml) and _{H 2 O (0.1ml), 1.25M} LiOH (70μL, 86μmol ) Was added and the mixture was stirred for 2.5 hours. After distillation under reduced pressure, the mixture was cooled to 0 ° C., 1.0 mL of MeOH was added, 0.5 M NaOH (50 μL) was added, and the mixture was stirred at room temperature for 16 days. 50% acetic acid (2 drops) was added and distilled off under reduced pressure, and the obtained residue was purified by gel filtration column chromatography (LH-20, 1% acetic acid) and Bond Elut (C8) to obtain 4.1 mg (207b) of 207b. (Yield 64%) was obtained. [α] _D + 18.3 ° (c 0.41, H ₂ O). ¹ H-NMR δ _H (H ₂ O) (DHO = 4.70 ppm): 7.00-6.99 (m, 2H, Ar), 6.87-6.85 (m) , 4H, Ar), 5.26 (d, 1H, J _1,2 = 3.78 Hz, H-1 ^4or2 ), 5.22 (d, 1H, J _1,2 = 3.84 Hz, H-1 ^2or4 ), 4.87 (d, 1H, J _1,2 = 7.80 Hz, H-1 ¹ ), 4.63 (d, 1H, J _1,2 = 8.00 Hz, H-1 ³ ), 3.68 (s, 3H, OMe), 3.61 (m, 1H , H-2 ^4or2 ), 3.56 (m, 2H, H-3 ³ , 3 ^2or4 ), 3.55 (m, 1H, H-2 ¹ ), 3.41 (dd, 1H, J _2,3 = 9.72 Hz, H- 2 ^2or4 ), 3.38 (dd, 1H, J _2,3 = 9.12 Hz, H-2 ³ ). ESI-HRMS m / z [(M + Na) ⁺ ]: calcd. For C ₂₉ H ₄₀ _{NaO 22} : 763.1903 Found, 763.1884.

Example 3 Synthesis of tetrasaccharide derivative shown in formula (9) (1) Synthesis of compound 402 Formula 10, compound 401 (273.6 mg, 273.6 mg, dissolved in a mixed solvent of toluene (6.0 mL) and ethanol (3.0 mL). H ₂ NNH ₂ · AcOH (112.1 mg) was added to (0.2516 mmol), and the mixture was stirred overnight at room temperature. Then, the reaction solution was concentrated and purified by gel filtration column chromatography (CHCl ₃ : MeOH = 1: 1) and silica gel column chromatography (ethyl acetate: methanol = 100: 1) to obtain 137.0 mg of compound 402 in yield. Obtained at 75%.

(2) Synthesis of Compound 403 (Formula 8) 10-Camphorsulfonic acid (3.9 mg) was added to Compound 402 (97.4 mg, 0.1340 mmol) dissolved in DMF, and the mixture was stirred at room temperature. 2-Methoxypropene (14 μL) was added thereto. After 8 hours, 2-methoxypropene (14 μL) was further added and the mixture was stirred overnight. Then, 2-methoxypropene (28 μL) was further added, and after 2 hours, diisopropylethylamine was added to stop the reaction, the mixture was extracted with EtOAc and dried over _{anhydrous sulfonyl 4.} The insoluble material was filtered, concentrated, and purified by silica gel column chromatography (toluene: ethyl acetate = 6: 1 to 4: 1) to obtain 70.2 mg (yield 68%) of compound 403 (formula 8).

(3) Synthesis of Compound 404 To MSAW300, Formula 7, Compound 205a (135.4 mg, 0.1948 mmol) and Compound 403 (110.0 mg, 0.1405 mmol) dissolved in _{CH 2} Cl _{2 were added.} The mixture was stirred at −78 ° C. TMSOTf (4.0 μL) was added thereto. After 1.5 hours, saturated NaHCO ₃ was added to stop the reaction, the mixture was extracted with _{CHCl 3} and dried over _{anhydrous plate 4.} The insoluble material was filtered, concentrated and purified by gel filtration column chromatography (CHCl ₃ : MeOH = 1: 1) to obtain a product (75.8 mg). _{Ac 2} O (3.0 mL) and pyridine (3.0 mL) were added to the obtained product, and the mixture was stirred overnight at room temperature. Then, the reaction mixture was concentrated and purified by silica gel column chromatography (toluene: ethyl acetate = 3: 1) to obtain 67.3 mg (2 step yield 35%) of compound 404.

(4) After activated with _{H 2} added THF (1.0 mL) in Synthesis (1,5-cyclooctadiene) bis (methyldiphenylphosphine ) iridium (I) hexafluorophosphate of Compound 405, it was Ar atmosphere. Compound 404 (67.3 mg, 0.04950 mmol) dissolved in THF (5.0 mL) was added thereto, and the mixture was stirred at room temperature for 1.5 hours. Then, water was added, the temperature was adjusted to 0 ° C., sodium hydrogen carbonate (8.9 mg) and iodine (26.2 mg) were added, and the mixture was stirred for 2 hours. Then, 1M hypo was added to stop the reaction, the mixture was extracted with _{CHCl 3} , and dried over _{anhydrous plate 4.} The insoluble material was filtered and concentrated, and purified by silica gel column chromatography (toluene: ethyl acetate = 3: 1 to ethyl acetate: methanol = 10: 1) to obtain a product (60.0 mg). The obtained product was dissolved in THF (4.0 mL), AcOH (25 μL) and 1 Mt-butylammonium fluoride (220 μL) were added, and the mixture was stirred at room temperature for 2 days. Then, it was extracted with _{CHCl 3} and dried with _{anhydrous plate 4.} The insoluble material was filtered and concentrated, and purified by silica gel column chromatography (toluene: ethyl acetate = 1: 1) to obtain 29.1 mg of compound 405 (2 step yield 54%).

(5) Synthesis of Compound 406 (Formula 9) 1.25 M LiOH (210 μL) was added to Compound 405 (29.1 mg, 0.0270 mmol) dissolved in a mixed solvent of THF (4.0 mL) and water (0.2 mL). The mixture was added at 0 ° C. and stirred for 1 hour. The reaction mixture was concentrated, methanol (4.0 mL) was added, 0.1 M NaOH (5 drops) was added, and the mixture was stirred overnight at room temperature. AcOH was added to stop the reaction, the reaction mixture was concentrated, and purified by gel filtration column chromatography (LH-20, 1% acetic acid) to obtain a product (20.0 mg). The obtained product was dissolved in a mixed solvent of methanol (2.0 mL) and water (2.0 mL), and palladium-carbon was added. Then, the flask with a H ₂ atmosphere and stirred at room temperature overnight. The reaction mixture was filtered through Celite, then concentrated and dried to obtain 14.6 mg of Compound 406 (formula 9, 92% yield in two steps).
¹ H-NMR δ _H (D ₂ O): 5.17 (d, 1H, J _1,2 = 3.84 Hz, Xyl ² -1), 4.44 (d, 1H, J _1,2 = 7.86 Hz, Xyl ¹ -1 ), 4.42 (d, 1H, J _1,2 = 8.22 Hz, GlcA-1), 3.94 (dd, 1H, J = 5.34, 11.88 Hz, Xyl-5a), 3.86-3.84 (m, 1H, Rbo-4) ), 3.74-3.72 (m, 4H, Xyl ² -4, Rbo-3, 5a, 1a), 3.69-3.60 (m, 5H, Xyl ¹ -4, GlcA-5, Rbo-2, 5b, Xyl ^2- 5a), 3.54-3.43 (m, 6H, Xyl ² -3, 5b, Xyl ¹ -3, GlcA-3,4, Rbo-1b), 3.38 (dd, 1H, J _2,3 = 9.66 Hz Xyl ^2- 2), 3.27 (d, 1H, J _2,3 = 9.12 Hz, GlcA-2), 3.24-3.20 (m, 2H, Xyl ¹ -2, Xyl ¹ -5b). ¹³ C NMR δ _C (D ₂ O) ^{): 105.46 (Xyl 1 -1)} , 103.84 (GlcA-1), 101.52 (Xyl 2 -1), 83.51 (GlcA-4), 83.33 (Rbo-4), 79.48 (Xyl 1 -4), 77.56 (Rbo -3), 76.37 (Xyl ¹ -3), 75.67 (Xyl ¹ -2), 75.60 (Xyl ² -3), 74.60, 74.40 (GlcA-5, Rbo-2), 74.20 (Xyl ² -2), 74.12 (Xyl ² -4), 73.85 (GlcA-2), 71.95 (GlcA-3), 65.53 (Xyl ¹ -5), 65.22 (Xyl ² -5), 64.03 (Rbo-1), 62.77 (Rbo-5) . ESI-HRMS m / z [(M + Na) ⁺ ]: calcd. For C ₂₁ H ₃₆ _{NaO 19} : 615.1748; found, 615.1744.

The present invention can be used in the fields of pharmaceutical manufacturing, sugar chain engineering, and the like.

Claims

Equation (1):

[In the formula, X 1 is a leaving group or a protected hydroxyl group, Y 1 is a carboxyl group or a hydroxymethyl group or a group that is a precursor thereof, and R 1 to R 5 are independently hydrogen. Is or is a hydroxyl-protecting group]
A disaccharide derivative represented by.
X 1 is a group selected from the group consisting of trichloroacetimideyloxy, alkylthio, arylthio, halogen, pentenyloxy, alkoxy and phenoxy, Y 1 is COOZ 1 or CH 2 OZ 2 , and Z 1 is an alkyl group. Z 2 is a protective group for hydroxyl groups, and R 1 to R 5 are independently substituted benzyl-containing benzyl, allyl, levulinoyl, substituted benzoyl-containing benzoyl, substituted acetyl-containing acetyl, allyloxycarbonyl, and isopropi. The disaccharide derivative according to claim 1, which is a group selected from the group consisting of lidene, benzylidene and trialkylsilyl.
Equation (2):

[In the formula, X 2 is a leaving group, Y is a group that fixes the configuration of the xylose residue, and R 6 is immobilized with p-alkyloxybenzyl, 3,4-dialkyloxybenzyl. P-alkyloxybenzyl, immobilized 3,4-dialkyloxybenzyl, naphthyl, naphthylmethyl, -CH = CH 2 , -C (CH 3 ) = CH 2 , -CH 2 -CH = CH 2 , -CH = CH-CH 3 , -CH = C = CH 2 , and a group selected from the group consisting of hydrogen]
With the xylose derivative represented by
Equation (3):

[In the formula, X 3 is a leaving group or a protected hydroxyl group, R 7 and R 8 are a protecting group for a hydroxyl group, and Y 4 is a group that is a precursor of a carboxyl group or a hydroxymethyl group].
From the glucose derivative or glucuronic acid derivative indicated by
Equation (4):

[In the equation, X 3 , R 7 , R 8 and Y 4 are the same as the above definitions]
A method for producing an α-glycoside represented by the following steps 1) to 3) :.
Step 1) By reacting the xylose derivative represented by the formula (2) with the glucose derivative or the glucuronic acid derivative represented by the formula (3), the OR 6 group of the xylose derivative and the glucose derivative or the glucuronic acid derivative are not protected. Manufacture mixed acetal derivatives via hydroxyl groups
Step 2) mixed acetal derivative obtained with step 1) is reacted to produce [alpha] 1 → 3 disaccharide derivative by activating the leaving group X 2, then,
Step 3) A method comprising deprotecting the disaccharide derivative obtained in Step 2) and inducing it into the formula (4).
X 2 is a group selected from the group consisting of trichloroacetimideyloxy, alkylthio, halogen, arylthio, and pentenyloxy, and Y is a group selected from the group consisting of acetal, carbonate, silylene acetal, and stanilen acetal. R 6 is a group selected from the group consisting of p-methoxybenzyl and naphthylmethyl, and X 3 is selected from the group consisting of trichloroacetimideyloxy, alkylthio, arylthio, halogen, pentenyloxy, alkoxy and phenoxy. R 7 and R 8 are independently selected from the group consisting of benzyl containing a substituted benzyl, allyl, lebrinoyl, benzoyl containing a substituted benzoyl, acetyl containing a substituted acetyl, allyloxycarbonyl and trialkylsilyl. The method according to claim 3, wherein Y 4 is COOZ 2 or CH 2 OZ 3 , Z 2 is an alkyl group, and Z 3 is a hydroxyl group protective group.
The method of claim 4, wherein R 7 is a group selected from the group consisting of benzoyls containing substituted benzoyls, acetyls containing substituted acetyls and levulinoyles.
Equation (2'):

Wherein, X 2 'is a leaving group, Y' is a group for fixing the conformation of xylose residues, R 6 'is a protecting group with no neighboring group participation ability hydroxyl]
With the xylose derivative represented by
Equation (3'):

Wherein, X 3 'is a leaving group or a protected hydroxyl group, R 7' is a protecting group for a hydroxyl group with a neighboring group participation ability, R 8 'is a protecting group for a hydroxyl group, Y 4' Is a group that is a precursor of a carboxyl or hydroxymethyl group]
From the glucose derivative or glucuronic acid derivative indicated by
Equation (4'):

Wherein, X 3 ', R 7' , R 8 ' and Y 4' are the same as defined above
A method for producing an α-glycoside represented by the following steps 1) to 3) :.
Step 1) The xylose derivative represented by the formula (2') and the glucose derivative or the glucuronic acid derivative represented by the formula (3') are directly condensed to obtain a mixture of α1 → 3 glycoside and β1 → 3 glycoside. ,
Step 2) Deprotect the mixture obtained in Step 1) and then
Step 3) A method comprising separating the α-glycoside from the deprotected mixture in Step 2).
The method according to claim 6, wherein the condensation in step 1) is carried out using NIS-AgOTf, NIS-TfOH, MeOTf, CuBr 2 -AgOTf-nBu 4 NI, or NBS-AgOTf as a condensing agent.
The deprotection of step 2) is to form triol having hydroxyl groups at the 2-, 3- and 4-positions of the xylose residue by hydrolysis using trifluoroacetic acid (TFA), claim 6 or 7. The method described.
The method according to any one of claims 6 to 8, wherein the separation in step 3) is performed by dissolving β-glycoside in chloroform to remove β-glycoside.
Formula (5): Containing the use of the disaccharide derivative represented by formula (1) as an intermediate (referred to as XG unit):

[In the formula, X 4 is a leaving group or a protected hydroxyl group, and n is an integer greater than or equal to 1.]
A method for producing an XG unit oligomer represented by.
The disaccharide derivative represented by the formula (1) is used as a sugar donor, and the formula (1'):

[In the formula, Y 5 is a group that is a precursor of a carboxyl group or a hydroxymethyl group, X 5 is a leaving group or a protected hydroxyl group, and R 9 to R 12 are a protecting group of a hydroxyl group].
The method according to claim 10, wherein the disaccharide derivative represented by the above is reacted as a sugar receptor.
Y 5 is COOZ 3 or CH 2 OZ 4 , Z 3 is an alkyl group, Z 4 is a hydroxyl group protective group, and X 5 is trichloroacetimideyloxy, alkylthio, arylthio, halogen, pentenyloxy, alkoxy. And a group selected from the group consisting of phenoxy, R 9 to R 12 are independently benzyl containing a substituted benzyl, allyl, levulinoyl, benzoyl containing a substituted benzoyl, acetyl containing a substituted acetyl, allyloxycarbonyl and 11. The method of claim 11, which is a group selected from the group consisting of trialkylsilyls.
12. The method of claim 12, wherein R 9 is a group selected from the group consisting of benzoyls containing substituted benzoyls, acetyls containing substituted acetyls and levulinoyles.
Equation (9):

Tetrasaccharides or derivatives thereof.
Equation (7):

[In the formula, X 1 is a leaving group or a protected hydroxyl group, X 2 to X 6 are hydroxyl protecting groups, and X 7 is an alkyl group].
The disaccharide donor represented by the formula (8):

Wherein, X 8 ~ X 13 is a hydroxyl-protecting group, nice contrast X 12 and X 13 may form acetal together]
The tetrasaccharide derivative is obtained by reacting with the disaccharide receptor represented by, and then the tetrasaccharide derivative is deprotected to obtain the formula (9) :.

A method for producing a tetrasaccharide represented by the formula (9), which comprises obtaining the tetrasaccharide represented by the formula (9).
X 1 is trichloroacetimideyloxy, X 2 to X 6 are acetyl, X 7 is methyl, X 8 is allyl, X 9 and X 10 are benzyl, X 11 is TBDPS, and X 12 and X 13 are together. The method of claim 15, wherein the isopropyridene acetal is formed.