CN118302451A

CN118302451A - Novel oligosaccharide, intermediate for producing the oligosaccharide, and process for producing the same

Info

Publication number: CN118302451A
Application number: CN202280072914.1A
Authority: CN
Inventors: 上田刚; 伊东龙生; 中村龙也; 铃木惠介; 中根智志; 杨泽堃
Original assignee: Daiichi Sankyo Co Ltd
Current assignee: Daiichi Sankyo Co Ltd
Priority date: 2021-10-29
Filing date: 2022-10-28
Publication date: 2024-07-05

Abstract

The problem of the present invention is to provide a novel oligosaccharide which can be used for producing a double-antennary glycan having an alpha 2, 6-sialic acid structure at the non-reducing end, a process for producing the same, and a process for producing the same, an intermediate and an intermediate thereof, wherein the double-antennary glycan has an alpha 2, 6-sialic acid structure at the non-reducing end. Provided are novel oligosaccharides represented by the following formulas A-13 and D-13, a process for producing the oligosaccharides represented by the formulas A-13 and D-13 shown in FIGS. 1 and 3, etc., and intermediates thereof and a process for producing the intermediates thereof.

Description

Novel oligosaccharide, intermediate for producing the oligosaccharide, and process for producing the same

Technical Field

The present invention relates to a novel oligosaccharide which is a double-antennary glycan having an α2, 6-sialic acid structure at the non-reducing end, a process for producing the oligosaccharide, an intermediate thereof, and a process for producing the intermediate.

Background

It is known that the addition of sugar chains (glycosylation) of proteins has a large influence on the function and structure of proteins. Among them, the N-linked sugar chain is closely related to the physiological activity of proteins, and among them, it is reported that a double-antennary N-glycan having an α2, 6-sialic acid structure at the non-reducing end is an optimal structure for improving antibody-dependent cellular cytotoxicity (ADCC activity) and complement-dependent cellular cytotoxicity (CDC activity) (non-patent document 1).

In the development and commercialization of pharmaceuticals using sugar chains, it is desired to stably produce sugar chains of high purity in large quantities at a commercially available price. The synthesis of the alpha 2, 6-sialylated sugar chain (. Alpha.2, 6-SIALYLGLYCAN) reported so far is largely divided into two methods: 1) Separation and purification of natural extracts, or semi-chemical synthesis combining chemical synthesis of main skeleton precursors with enzymatic chemical conversion; 2) And (3) a pure chemical synthesis method.

For example, as a semi-chemical synthesis method, it has been reported that an N-linked sugar chain can be obtained from the yolk of an egg by combining a method using an enzyme with a chemical method (non-patent document 2). These methods can synthesize a target sugar chain in a smaller number of steps than the pure chemical synthesis, and on the other hand, a large amount of egg yolk is required, and in the subsequent isolation and purification from egg yolk and purification of a water-soluble unprotected sugar chain after chemical conversion, a special technique and a purification apparatus are often required (patent documents 1 to 4).

On the other hand, the following examples are reported as the pure synthesis method of the α 2, 6-sialylated sugar chain.

(1) Total synthesis of complex 11-type glycans having an α2, 6-sialic acid moiety (non-patent document 3).

(2) Total synthesis of immunoglobulin G13 glycopeptides having an α2, 6-sialic acid moiety (non-patent document 4).

(3) Total synthesis of an α 2, 6-sialylated 12-sugar N-linked sugar chain containing core fucose (non-patent document 5).

(4) Total synthesis of a2, 6-sialylated 10-oligosaccharide chain with fluorinated 3-position (non-patent document 6).

(5) Complete synthesis of asymmetric deuterated α2, 6-sialylated double antennary 11 sugar oligosaccharide chain and tetraantennary 17 sugar oligosaccharide chain (non-patent document 7)

It is considered that, when a reliable production method is established, the pure synthesis of sugar chains is derived from monosaccharides, similarly to a general low-molecular compound, and the degree of freedom in the number of production is extremely high. Further, since the conversion of the sugar modified with the protecting group is performed, most of the purification operation is performed as a treatment of the water-insoluble compound, and thus, it is expected that the complexity of the operation and the man-hour of the operation are significantly reduced as compared with the semi-chemical synthesis method. Furthermore, by applying the established chemical synthesis method, various unnatural sugar chains can be easily synthesized.

On the other hand, in the above-described conventional report, the following two points are listed as major technical problems in synthesis: 1) The conversion of sugar moieties with high difficulty, such as the construction of β -mannoside moieties and α -sialic acid moieties, is also carried out in a step of connecting them, and there is a step of low selectivity and low yield; 2) In both the sugar partial conversion and the linking step, silica gel column chromatography purification which is unsuitable for scale-up is often used in each step, and in many steps, a precise chromatographic separation and purification operation is required for the purpose of removing isomers and impurities which are by-produced in the reaction.

As described above, the pure chemical synthesis method of sugar chains has potential advantages over the semi-chemical synthesis method in terms of mass synthesis, but it is difficult to say that the technology is developed in a state of sufficient progress from the viewpoints of yield, selectivity, efficiency and cost, and there are few examples of the mass synthesis method used as a practical one. Furthermore, it is very difficult to structurally perform a large amount of synthesis of double-antennary sugar chains (. Alpha.2, 6-sialylated sugar chains) in which different sugar chain units are linked to the 3,6 positions of mannose at branching points, in all of derivatization, semi-chemical synthesis and total chemical synthesis from natural extracts.

In addition, in the sugar derivative protected by a phthalimido group, it is sometimes necessary to simultaneously benzyl a plurality of hydroxyl groups. In this case, it is necessary to suppress the ring opening of the phthalimide group and allow the reaction to proceed, but the phthalimide group is likely to undergo the ring opening reaction under strongly basic conditions due to the presence of a small amount of hydroxide ion, and therefore under the NaH/DMAc conditions used in the conventional benzylation reaction, the yield greatly varies depending on the amount of sodium hydroxide in NaH. In addition, naH/DMAc is not easy to be applied to a large number of synthesis methods due to the danger of mixing and explosion hazards. Thus, a method of inhibiting the ring opening of phthalimide groups and simultaneously benzylating a plurality of hydroxyl groups under milder conditions is demanded.

Furthermore, sugar derivatives protected by phthalimido groups sometimes require deacylation. However, when a trace amount of moisture is present in the system, since the phthalimide group is liable to undergo a ring-opening reaction under alkaline conditions, it is necessary to strictly control the moisture content in the system, and it is difficult to completely suppress the ring opening even at a moisture value of ppm level. Therefore, a method of suppressing the ring opening of phthalimide groups and enabling deacylation to be performed in high yield is demanded.

In chemical synthesis of oligosaccharide chains having a hydroxyl group(s) and the like, a method of selectively protecting and deprotecting the hydroxyl group is required to be reasonably utilized in order to obtain a target compound efficiently, and particularly 2-naphthylmethyl has been widely used as a general protecting group for a hydroxyl group. On the other hand, as a deprotection method of 2-naphthylmethyl group, a method of using 2, 3-dichloro-5, 6-dicyano-p-benzoquinone in methylene chloride-water is known, which provides a target deprotected body in a moderate to high yield for a wide range of substrates, but since methylene chloride is generally not miscible with water, and 2, 3-dichloro-5, 6-dicyano-p-benzoquinone (benzohydroquinone) as a by-product thereof are hardly dissolved in methylene chloride-water, and thus have an influence on stirring property, there is a problem in application to mass synthesis. The following cases were also reported: depending on the substrate used, the target deprotected substance is not necessarily obtained in a satisfactory yield (non-patent document 8). Further, since benzyl groups in the matrix tend to be increased, the reaction yield tends to be lowered (non-patent document 9), it is desired to develop a milder and efficient method that can be applied to a complex matrix. As a method for achieving the above object, an improved condition using β -pinene as an additive is reported (non-patent document 10), but even if this method is used, the yield stays at a moderate level for a complex substrate having a plurality of benzyl groups (non-patent document 11). In view of the above background, it is desired to develop a deprotection reaction for deprotecting a protecting group of a 2-naphthylmethyl group under milder conditions, and a deprotected 2-naphthylmethylation reaction for deprotecting a body can be obtained in high yield.

Further, as a chemical synthesis method of an oligosaccharide chain, a liquid phase synthesis method and a solid phase synthesis method are known. However, since the liquid phase synthesis method can use a general organic synthesis method, it is easy to track the reaction and scale up, but it is time-consuming and labor-consuming to carry out the post-treatment and purification at each step, and furthermore, the solid phase synthesis method is advantageous in that it can be automated and can be produced rapidly, but there is a limit in scale up due to equipment limitations, and furthermore, it is necessary to use the sugar donor used in the sugar extension reaction in excess because of low reactivity, and it is not suitable for industrial mass synthesis, and furthermore, there is a disadvantage that it is difficult to confirm the progress of the reaction in the middle stage (patent document 5). In order to solve these problems, several methods for producing oligosaccharides using a substrate to which a molecular structure (tag) is attached, which specifically causes precipitation, partition, adsorption, and the like, to a certain environment, have been developed (patent document 6, non-patent documents 12, 13, and 14). These methods have the advantage of both liquid-phase and solid-phase synthesis methods, that is, they can be easily analyzed by performing the reaction in a homogeneous system, and can be separated from reagent residues and the like by utilizing the characteristics of the tag. For example, the following methods are known: the branched long-chain alkane is used as a hydrophobic tag, and the solution after the reaction is adsorbed on octadecyl-modified silica gel, whereby the unlabeled compound is separated (non-patent document 14). However, the labels must be used in either method, which has the following disadvantages: a desorption process is required; and, as oligomerization proceeds, the matrix site becomes larger than the tag, so that physical properties of the matrix gradually become dominant and functions of the tag become weaker. Therefore, in the method for producing an oligosaccharide chain, a method capable of purifying an oligosaccharide more efficiently is demanded.

In the case where the-NHAc group is present in the reaction matrix during the glycosylation reaction, it can be found that the reactivity of the target glycosylation reaction is significantly reduced due to interaction with the Lewis acid, in most cases an excess of sugar donor is required for completion of the reaction. Thus, in the case of synthesizing an oligosaccharide chain containing acetylglucosamine, the following method is employed: in the glycosylation reaction, a Troc group (non-patent documents 5, 6 and 7), a phthalimide group, or a sulfonyl group (non-patent document 4) is used as a temporary protecting group on the nitrogen of glucosamine, and then deprotected and then subjected to N-Ac conversion. However, in the case of the Troc group, deprotection reaction conditions based on zinc/AcOH or long-time reaction based on excessive lithium hydroxide are required, and complex sugar chains accompany the decomposition of the matrix under the deprotection reaction conditions. In addition, since ethylenediamine is used in excess in deprotection of phthalimide group, there is a problem that amidation of sialic acid ester site is performed by side reaction, and in order to avoid this, it is necessary to carry out treatment of two steps of selective hydrolysis of ester site in advance and deprotection. Furthermore, in the case of sulfonyl groups, deprotection is performed under reaction conditions where scale up is difficult to achieve using metallic sodium. Therefore, in the method for producing an oligosaccharide chain containing acetylglucosamine, development of a protecting group which does not reduce the reactivity of glycosylation reaction and can easily replace an acetyl group has been demanded.

Polyethylene glycol has been widely used in recent years as a biocompatible water-soluble partial structure for development of pharmaceuticals and the like. In the development of such a pharmaceutical product, it is desired that these polyethylene glycols have a more uniform structure and a higher purity, but various impurities are mixed in a commercially available reagent, and further, the purification thereof requires the implementation of strict distillation purification or complicated column purification. In addition, in the case where the compound having a polyethylene glycol structure contains an azide structure, a distillation operation such as heating is not applicable due to its explosion hazard. Recently, a method of purifying a metal complex using MgCl ₂ has been reported (non-patent document 15), but since the target substance is adsorbed to MgCl ₂ in excess, the loss in the filtrate is large, and the purification effect is predicted to be smaller than that of isolation by crystallization. Accordingly, a method for purifying a compound having the polyethylene glycol structure is desired.

In the synthesis of oligosaccharide chains, as the molecular weight increases, isolation and purification of the intermediate in the form of crystals becomes difficult. In particular, no examples have been reported of crystallization of an intermediate having a molecular weight of more than 1000 and being a protected trisaccharide. Therefore, how to remove from the target substance an analogue having a similar structure, such as an isomer, an impurity derived from the residual raw material, and the like, which are by-produced in a trace amount. Conventionally, a method of purifying a silica gel column in each step of synthesis has been employed for the purpose of removing these analogues, and this has been a major obstacle in achieving efficient mass synthesis. In view of the above background, development of a crystallization and purification method capable of efficiently removing impurities having a similar structure from an oligosaccharide chain synthesis intermediate has been desired.

Prior art literature

Patent literature

Patent document 1: international publication No. 2011/027868

Patent document 2: international publication No. 96/02255

Patent document 3: international publication No. 2014/208742

Patent document 4: international publication No. 2017/110984

Patent document 5: international publication No. 2002/16384

Patent document 6: japanese patent No. 6001267 specification

Non-patent literature

Non-patent document 1: proc.Natl.Acad.Sci.U.S.A.2015, 112, 10611-10616

Non-patent document 2: beilsteinJ.org.chem.2018, 14, 416-429

Non-patent document 3: tetrahedronLett.1986, 27, 5739-5742

Non-patent document 4: j.am.chem.soc.2009, 131, 16669-16671

Non-patent document 5: j.org.chem.2016, 81, 10600-10616

Non-patent document 6: J.am.chem.Soc.2019, 141, 6484-6488

Non-patent document 7: angew.chem.int.ed.2021, 60, 24686-24693

Non-patent document 8: org. Lett.2002,4, 4551-4554

Non-patent document 9: J.am.chem.Soc.2018, 140, 4632-4638

Non-patent document 10: org.chem.2017, 82, 3926-3934

Non-patent document 11: angew.chem.int.ed.2021, 60, 19287-19296

Non-patent document 12: organic synthesis society, volume 60, 2002, no. 5, p.494-495

Non-patent document 13: org.biomol.chem.2018, 16, 4720-4727

Non-patent document 14: J.AM.CHEM.SOC.2005, 127, 7296-7297

Non-patent document 15: org.Process.Res.Dev.2021, 25, 10, 2270-2276

Disclosure of Invention

Problems to be solved by the invention

One problem of the present invention is to provide a novel oligosaccharide which can be used for producing a double-antenna glycan having an α2, 6-sialic acid structure at the non-reducing end, a process for producing the oligosaccharide, an intermediate thereof, and a process for producing the intermediate. Another problem of the present invention is to provide a novel oligosaccharide which is a double-antennary glycan having an α2, 6-sialic acid structure at the non-reducing end, a process for producing the oligosaccharide, an intermediate thereof, and a process for producing the intermediate.

Solution for solving the problem

The present inventors have conducted intensive studies to solve the above problems, and as a result, found that: a novel oligosaccharide represented by the following A-13, a novel method for producing the oligosaccharide with high efficiency, an intermediate thereof, and a method for producing the intermediate; the present invention has been completed as a result of the above-described finding, a novel oligosaccharide represented by D-13 below as a double-antenna glycan having an α2, 6-sialic acid structure at the non-reducing end, a novel method for producing the oligosaccharide with high efficiency, and a method for producing an intermediate thereof and an intermediate thereof.

That is, the present application relates to the following, but is not limited thereto.

[1] A method for producing an oligosaccharide represented by the following formula A-13.

The method comprises a working procedure I-1, a working procedure I-2 and a working procedure I-3.

(Step I-1) is a step of producing a compound represented by the following formula A-7, and includes the following steps: the compound represented by the following formula A-5 is produced by reacting a compound represented by the formula A-3 with a compound represented by the following formula A-4 to form an alpha-1, 6-glycosidic bond.

(Step I-2) is a step of producing a compound represented by the following formula A-10, and includes the following steps: the compound represented by the following formula A-9 is produced by forming a beta-1, 4-glycosidic bond between the compound represented by the formula A-7 and the compound represented by the following formula A-8.

(Step I-3) is a step of producing the oligosaccharide represented by the formula A-13, and comprises the steps of: the compound represented by the following formula A-12 is produced by forming a beta-1, 2-glycosidic bond between the compound represented by the formula A-10 and the compound represented by the following formula A-11.

[2] The method according to [1], wherein the step I-2 includes: the compound represented by the formula A-10 is produced by reacting the compound represented by the formula A-9 with a strong base in the presence of an alkyl ester of a perfluorocarboxylic acid.

[3] The method according to [2], wherein the alkyl ester of perfluorocarboxylic acid is methyl trifluoroacetate, ethyl trifluoroacetate, propyl trifluoroacetate, isopropyl trifluoroacetate, butyl trifluoroacetate, methyl pentafluoropropionate, ethyl pentafluoropropionate, propyl pentafluoropropionate, isopropyl pentafluoropropionate, methyl heptafluorobutyrate, ethyl heptafluorobutyrate, propyl heptafluorobutyrate, isopropyl heptafluorobutyrate, butyl heptafluorobutyrate, methyl nonafluoropentanoate, ethyl nonafluoropentanoate, propyl nonafluoropentanoate, isopropyl nonafluoropentanoate, butyl nonafluoropentanoate, methyl undecanoate, ethyl undecanoate, propyl undecanoate, isopropyl undecanoate, or butyl undecanoate.

[4] The method of [2] or [3], wherein the strong base is selected from the group consisting of: sodium, lithium, and potassium salts of metal amides; sodium, lithium, potassium, cesium, and barium salts of C1-C20 alkoxides; sodium hydride, potassium hydride, lithium hydride, butyllithium, potassium carbonate, sodium carbonate, cesium carbonate, lithium carbonate, potassium phosphate, sodium phosphate, cesium phosphate, lithium phosphate, diazabicycloundecene (DBU), diazabicyclononene (DBN), and 1, 3-Tetramethylguanidine (TMG); and combinations thereof.

[5] The method according to [2] or [3], wherein the strong base is potassium tert-butoxide, sodium tert-butoxide, lithium tert-butoxide, or LHMDS (lithium hexamethyldisilazide).

[6] The method according to any one of [2] to [5], wherein the reaction of step I-2 is performed in a C1-C10 alcohol solvent alone or in a mixed solvent of a C1-C10 alcohol solvent and an amide-based solvent, an ether-based solvent, an ester-based solvent, an aromatic-based solvent, a halogen-based solvent, a hydrocarbon-based solvent, or a nitrile-based solvent.

[7] The method according to any one of [1] to [6], wherein the step I-3 includes: the compound represented by the formula A-12 is reacted with DDQ (2, 3-dichloro-5, 6-dicyano-p-benzoquinone) in a mixed solvent of a fluoroalcohol and water to disengage the 2-naphthylmethyl group in the compound represented by the formula A-12, thereby producing the oligosaccharide represented by the formula A-13.

[8] The method according to [7], wherein, the fluoroalcohol is selected from the group consisting of hexafluoro-2-propanol (HFIP), 2-Trifluoroethanol (TFE) 2,3, 4, 5-octafluoro-1-pentanol, nonafluoro-tert-butanol, and combinations thereof.

[9] The method according to [7] or [8], wherein the reaction of the step I-3 is carried out at-35℃to 70 ℃.

[10] The method according to [7] or [8], wherein the reaction of the step I-3 is carried out at-30℃to-10 ℃.

[11] The method according to any one of [1] to [10], wherein the step I-1 includes: after stopping the reaction between the compound represented by the formula A-4 and the compound represented by the formula A-3, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the compound represented by the formula A-5 and inclusions formed, the compound represented by the formula A-5 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and the water, whereby the inclusions are removed, and then the compound represented by the formula A-5 is eluted from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula A-5.

[12] The method according to any one of [1] to [11], wherein the step I-2 includes: after stopping the reaction between the compound represented by the formula A-7 and the compound represented by the formula A-8, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the produced compound represented by the formula A-9 and inclusions, the compound represented by the formula A-9 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and the water, whereby the inclusions are removed, and then the compound represented by the formula A-9 is eluted from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula A-9.

[13] The method according to any one of [1] to [12], wherein the step I-3 includes: after stopping the reaction between the compound represented by the formula A-10 and the compound represented by the formula A-11, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the compound represented by the formula A-12 and inclusions formed, the compound represented by the formula A-12 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and the water, whereby the inclusions are removed, and then the compound represented by the formula A-12 is eluted from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula A-12.

[14] The method according to [11], wherein the inclusion contains a sugar compound other than the compound represented by the formula A-5 and/or a compound derived from a reactant for obtaining the purified compound.

[15] The method according to [12], wherein the inclusion contains a sugar compound other than the compound represented by the formula A-9 and/or a compound derived from a reactant for obtaining the purified compound.

[16] The method according to [13], wherein the inclusion contains a sugar compound other than the compound represented by the formula A-12 and/or a compound derived from a reactant for obtaining the purified compound.

[17] The method according to any one of [11] to [16], wherein the hydrophobic carrier is a resin for reversed-phase partition chromatography packing.

[18] The method of [17], wherein the reverse phase partition chromatography packing resin is selected from the group consisting of poly (styrene/divinylbenzene) polymer gel resins, polystyrene-divinylbenzene resins, polyhydroxymethacrylate resins, styrene-vinylbenzene copolymer resins, polyvinyl alcohol resins, polystyrene resins, polymethacrylate resins, chemically bonded silica gel resins, and combinations thereof.

[19] The method according to [18], wherein the chemically bonded silica gel resin is selected from the group consisting of: (1) A resin obtained by reacting a silane coupling agent with silica gel; (2) A resin obtained by chemically bonding dimethyloctadecyl, octadecyl, trimethyloctadecyl, dimethyloctyl, octyl, butyl, ethyl, methyl, phenyl, cyanopropyl, or aminopropyl to silica gel; (3) A resin obtained by chemically bonding a behenyl group or a triacontyl group to silica gel; and (4) combinations of the above (1) to (3).

[20] The method according to [18], wherein the chemically bonded silica gel resin is an octadecyl bonded silica gel resin (ODS resin).

[21] The method according to any one of [11] to [20], wherein the water-soluble organic solvent is a water-soluble alcohol-based solvent, a water-soluble nitrile-based solvent, a water-soluble ether-based solvent, a water-soluble ketone-based solvent, a water-soluble amide-based solvent, or a water-soluble sulfoxide-based solvent, or a mixed solvent comprising at least one or more of the above water-soluble organic solvents.

[22] The method according to [21], wherein the water-soluble nitrile solvent is acetonitrile.

[23] The method according to any one of [11] to [22], wherein the organic solvent used in the step of eluting the target from the hydrophobic carrier is a nitrile-based solvent, an ether-based solvent, an ester-based solvent, a ketone-based solvent, a halogen-based solvent, an aromatic-based solvent, or a mixed solvent comprising at least one of the above solvents.

[24] The method according to any one of [1] to [23], wherein the compound represented by the formula A-11 is produced by a process comprising a process Y-1 and a process Y-2.

(Step Y-1) is a step of producing a compound represented by the following formula B-4, and includes the steps of: the compound represented by the following formula B-3 is produced by forming a beta-1, 4-glycosidic bond between a compound represented by the following formula B-1 and a compound represented by the following formula B-2.

The (step Y-2) is as follows: adding lithium t-butoxide or lithium t-amyl alcohol to a solvent comprising the compound represented by the formula B-4 and benzyl halide or benzyl sulfonate, protecting the hydroxyl group existing in the compound represented by the formula B-4 with benzyl, thereby generating a compound represented by the following formula B-5,

[25] The method according to [24], wherein the solvent comprising the compound represented by the formula B-4 and benzyl halide or benzyl sulfonate is an amide-based solvent, an ether-based solvent, an aromatic-based solvent, a hydrocarbon-based solvent, a urea-based solvent, or a mixed solvent comprising at least one of the above solvents.

[26] The method according to [24] or [25], wherein the method further comprises the steps of: the compound represented by the formula B-5 is prepared by opening the ring of the phthalimide group in the compound represented by the formula B-5, then forming a salt with cinchonidine to produce a crystalline compound represented by the following formula B-6, separating the crystalline compound represented by the formula B-6 from an amorphous substance, then removing cinchonidine in the compound represented by the formula B-6 by adding an acidic aqueous solution and a solvent to produce a compound represented by the following formula B-7, and then closing the ring of the ring-opened phthalimide group in the compound represented by the formula B-7 to purify the compound represented by the formula B-5.

[27] The method according to any one of [1] to [26], further comprising the steps of: the compound represented by the formula A-13 is purified by opening the phthalimide group in the compound represented by the formula A-13, then forming a salt with (R) - (+) -1- (1-naphthyl) ethylamine to produce a crystalline compound represented by the following formula A-14, separating the crystalline compound represented by the formula A-14 from an amorphous substance, removing the (R) - (+) -1- (1-naphthyl) ethylamine in the compound represented by the formula A-14 by adding an acidic aqueous solution and a solvent to produce a compound represented by the following formula A-15, and then closing the ring of the phthalimide group ring-opened in the compound represented by the formula A-15.

[28] A method for producing an oligosaccharide represented by the following formula D-13.

The method comprises a working procedure II-1, a working procedure II-2, a working procedure II-3 and a working procedure II-4.

(Step II-1) is a step of producing a compound represented by the following formula D-2, and includes the following steps: the oligosaccharide represented by the following formula A-13 and the compound represented by the following formula A-3 are formed into an alpha-1, 3-glycosidic bond to produce a compound represented by the following formula D-1.

(Step II-2) is the following step: after the compound represented by the following formula D-5 is produced, the amino group in the compound represented by the formula D-5 is protected with a protecting group selected from the group consisting of an aryloxycarbonyl group (COOAr), an acetyl group (Ac), a 2, 2-trichloroethoxycarbonyl group (Troc), and a phthalimide group (Pht), to produce the compound represented by the following formula D-6 (wherein R ₅ is an aryloxycarbonyl group (COOAr), an acetyl group (Ac), or a 2, 2-trichloroethoxycarbonyl group (Troc), and R ₆ is a hydrogen atom, or R ₅ and R ₆ form a phthalimide group together with the nitrogen atom to which they are bonded); or removing acetyl (Ac) groups from the compound represented by the following formula D-4 to form a compound represented by the following formula D-6 (wherein R ₅ and R ₆ form a phthalimide group together with the nitrogen atom to which they are bonded), and the step II-2 comprises a step of forming a beta-1, 2-glycosidic bond between the compound represented by the formula D-2 and the compound represented by the following formula D-3 to form a compound represented by the following formula D-4.

(Step II-3) is a step of producing a compound represented by the following formula D-11 (wherein M ⁺ is sodium ion, lithium ion, potassium ion, or protonated triethylamine cation), and comprises the steps of: by forming β -1, 4-glycosidic bond between the compound represented by the formula D-6 and the compound represented by the formula D-7 below, a compound represented by the formula D-8 below is produced (wherein R ₅ is aryloxycarbonyl (COOAr), acetyl (Ac), or 2, 2-trichloroethoxycarbonyl (Troc), and R ₆ is hydrogen atom, or R ₅ and R ₆ form phthalimide group together with the nitrogen atom to which they are bonded), and then the protecting group of the acyl protecting group of the amino group and alcohol on the compound represented by the formula D-8 is removed, to produce a compound represented by the formula D-9 below (wherein M ⁺ is sodium ion, lithium ion, potassium ion, or protonated triethylamine cation).

(Step II-4) is the following step: the oligosaccharide represented by the formula D-13 is produced by reacting the compound represented by the formula D-11 with a compound represented by the following formula D-12.

[29] The method according to [28], wherein the step II-1 comprises: the compound represented by the formula D-2 is produced by reacting the compound represented by the formula D-1 with a strong base in the presence of an alkyl ester of a perfluorocarboxylic acid.

[30] The method according to [29], wherein the alkyl ester of perfluorocarboxylic acid is methyl trifluoroacetate, ethyl trifluoroacetate, propyl trifluoroacetate, isopropyl trifluoroacetate, butyl trifluoroacetate, methyl pentafluoropropionate, ethyl pentafluoropropionate, propyl pentafluoropropionate, isopropyl pentafluoropropionate, methyl heptafluorobutyrate, ethyl heptafluorobutyrate, propyl heptafluorobutyrate, isopropyl heptafluorobutyrate, butyl heptafluorobutyrate, methyl nonafluoropentanoate, ethyl nonafluoropentanoate, propyl nonafluoropentanoate, isopropyl nonafluoropentanoate, butyl nonafluoropentanoate, methyl undecanoate, ethyl undecanoate, propyl undecanoate, isopropyl undecanoate, or butyl undecanoate.

[31] The method of [29] or [30], wherein the strong base is selected from the group consisting of: sodium, lithium, and potassium salts of metal amides; sodium, lithium, potassium, cesium, and barium salts of C1-C20 alkoxides; sodium hydride, potassium hydride, lithium hydride, butyllithium, potassium carbonate, sodium carbonate, cesium carbonate, lithium carbonate, potassium phosphate, sodium phosphate, cesium phosphate, lithium phosphate, diazabicycloundecene (DBU), diazabicyclononene (DBN), and 1, 3-Tetramethylguanidine (TMG); and combinations thereof.

[32] The method of [29] or [30], wherein the strong base is potassium tert-butoxide, sodium tert-butoxide, lithium tert-butoxide, or LHMDS (lithium hexamethyldisilazide).

[33] The method according to any one of [29] to [32], wherein the step of producing the compound represented by the formula D-2 by reacting the compound represented by the formula D-1 with a strong base in the presence of trifluoroacetate is performed in a C1-C10 alcohol solvent alone or in a mixed solvent of a C1-C10 alcohol solvent and an amide-based solvent, an ether-based solvent, an ester-based solvent, an aromatic-based solvent, a halogen-based solvent, a hydrocarbon-based solvent, or a nitrile-based solvent.

[34] The method according to any one of [28] to [33], wherein in the step II-3, the compound represented by the formula D-6 is produced by protecting an amino group in the compound represented by the formula D-5 with an aryloxycarbonyl group (COOAr).

[35] The method according to any one of [28] to [34], wherein in the step II-3, the step of producing the compound represented by the formula D-6 from the compound represented by the formula D-5 is performed in an aqueous solution of sodium hydrogencarbonate, potassium hydrogencarbonate, disodium hydrogenphosphate, or dipotassium hydrogenphosphate.

[36] The method according to any one of [28] to [35], wherein the compound represented by the formula D-12 is obtained by a purification method comprising the steps of: a step of adding a compound represented by the following formula E-1 (wherein R ₇ is a hydrogen atom, a methyl group, or a methoxy group) to a solution containing the crude compound represented by the formula D-12 to produce a crystalline compound represented by the following formula E-2 (wherein R ₇ is a hydrogen atom, a methyl group, or a methoxy group); and isolating the crystalline compound, followed by a step of extracting the compound represented by the formula D-12 from the isolated crystalline compound.

[37] The method according to [36], wherein the compound represented by the formula D-12 after purification has a purity of 95% or more as measured by HPLC.

[38] The method according to [37], wherein the purity is 98% or more.

[39] The method according to any one of [28] to [38], wherein the step II-1 includes: after stopping the reaction between the compound represented by the formula A-13 and the compound represented by the formula A-3, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the compound represented by the formula D-1 and inclusions formed, the compound represented by the formula D-1 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and the water, whereby the inclusions are removed, and then the compound represented by the formula D-1 is eluted from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula D-1.

[40] The method according to any one of [28] to [39], wherein the step II-2 includes: after stopping the reaction between the compound represented by the formula D-3 and the compound represented by the formula D-4, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the compound represented by the formula D-5 and inclusions formed, the compound represented by the formula D-5 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and the water, whereby the inclusions are removed, and then the compound represented by the formula D-5 is eluted from the hydrophobic carrier using an organic solvent, whereby the compound represented by the formula D-5 is purified.

[41] The method according to any one of [28] to [40], wherein the step II-3 includes: after stopping the reaction between the compound represented by the formula D-6 and the compound represented by the formula D-7, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the compound represented by the formula D-8 and inclusions formed, the compound represented by the formula D-8 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and the water, whereby the inclusions are removed, and then the compound represented by the formula D-8 is eluted from the hydrophobic carrier using an organic solvent, whereby the compound represented by the formula D-8 is purified.

[42] The method according to any one of [28] to [41], wherein the step II-3 includes: the compound represented by the formula D-10 is purified by forming a compound represented by the following formula D-10 by protecting an amino group on the formula D-9 with an acetyl group, adding a hydrophobic carrier and water to a water-soluble organic solvent containing the formed compound represented by the formula D-10 and an inclusion, adsorbing the compound represented by the formula D-10 to the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and the water, thereby removing the inclusion, and eluting the compound represented by the formula D-10 from the hydrophobic carrier using an organic solvent.

[43] The method according to [39], wherein the inclusion contains a sugar compound other than the compound represented by the formula D-1, and/or a compound derived from a reactant for obtaining the purified compound.

[44] The method according to [40], wherein the inclusion contains a sugar compound other than the compound represented by the formula D-5, and/or a compound derived from a reactant for obtaining the purified compound.

[45] The method according to [41], wherein the inclusion contains a sugar compound other than the compound represented by the formula D-8, and/or a compound derived from a reactant for obtaining the purified compound.

[46] The method according to [42], wherein the inclusion contains a sugar compound other than the compound represented by the formula C-10, and/or a compound derived from a reactant for obtaining the purified compound.

[47] The method according to any one of [39] to [46], wherein the hydrophobic carrier is a resin for reversed-phase partition chromatography packing.

[48] The method of [47], wherein the reverse phase partition chromatography packing resin is selected from the group consisting of poly (styrene/divinylbenzene) polymer gel resins, polystyrene-divinylbenzene resins, polyhydroxymethacrylate resins, styrene-vinylbenzene copolymer resins, polyvinyl alcohol resins, polystyrene resins, polymethacrylate resins, chemically bonded silica gel resins, and combinations thereof.

[49] The method according to [48], wherein the chemically bonded silica gel resin is selected from the group consisting of: (1) A resin obtained by reacting a silane coupling agent with silica gel; (2) A resin obtained by chemically bonding dimethyloctadecyl, octadecyl, trimethyloctadecyl, dimethyloctyl, octyl, butyl, ethyl, methyl, phenyl, cyanopropyl, or aminopropyl to silica gel; (3) A resin obtained by chemically bonding a behenyl group or a triacontyl group to silica gel; and (4) combinations of the above (1) to (3).

[50] The method according to [48], wherein the chemically bonded silica gel resin is an octadecyl bonded silica gel resin (ODS resin).

[51] The method according to any one of [39] to [50], wherein the water-soluble organic solvent is a water-soluble alcohol-based solvent, a water-soluble nitrile-based solvent, a water-soluble ether-based solvent, a water-soluble ketone-based solvent, a water-soluble amide-based solvent, or a water-soluble sulfoxide-based solvent, or a mixed solvent comprising at least one or more of the above water-soluble organic solvents.

[52] The method according to [51], wherein the water-soluble nitrile solvent is acetonitrile.

[53] The method according to any one of [39] to [52], wherein the organic solvent used in the step of eluting the target from the hydrophobic carrier is a nitrile-based solvent, an ether-based solvent, an ester-based solvent, a ketone-based solvent, a halogen-based solvent, an aromatic-based solvent, or a mixed solvent comprising at least one of the above solvents.

[54] The method according to any one of [28] to [53], further comprising the steps of: the compound represented by the formula D-5 is salified with fumaric acid to form a crystalline compound represented by the following formula D-5-FMA, and then the crystalline compound represented by the formula D-5-FMA is separated from an amorphous substance and purified.

[55] A method for producing a compound represented by the following formula B-5.

The method comprises the following steps: the hydroxyl group present in the compound represented by the formula B-4 is protected with a benzyl group by adding lithium t-butoxide or lithium t-amyl alcohol to a solvent comprising the compound represented by the formula B-4 and a benzyl halide or benzyl sulfonate.

[56] The method according to [55], wherein the solvent is an amide-based solvent, an ether-based solvent, an aromatic-based solvent, a urea-based solvent, a hydrocarbon-based solvent, or a mixed solvent comprising at least one or more of the above solvents.

[57] A method for producing a compound represented by the following formula a-10, comprising the steps of: the compound represented by the following formula A-9 is reacted with a strong base in the presence of an alkyl ester of perfluorocarboxylic acid.

[58] The method according to [57], wherein the alkyl ester of perfluorocarboxylic acid is methyl trifluoroacetate, ethyl trifluoroacetate, propyl trifluoroacetate, isopropyl trifluoroacetate, butyl trifluoroacetate, methyl pentafluoropropionate, ethyl pentafluoropropionate, propyl pentafluoropropionate, isopropyl pentafluoropropionate, methyl heptafluorobutyrate, ethyl heptafluorobutyrate, propyl heptafluorobutyrate, isopropyl heptafluorobutyrate, butyl heptafluorobutyrate, methyl nonafluoropentanoate, ethyl nonafluoropentanoate, propyl nonafluoropentanoate, isopropyl nonafluoropentanoate, butyl nonafluoropentanoate, methyl undecanoate, ethyl undecanoate, propyl undecanoate, isopropyl undecanoate, or butyl undecanoate.

[59] The method of [57] or [58], wherein the strong base is selected from the group consisting of: sodium, lithium, potassium salts of metal amides; sodium, lithium, potassium, cesium, and barium salts of C1-C20 alkoxides; sodium hydride, potassium hydride, lithium hydride, butyllithium, potassium carbonate, sodium carbonate, cesium carbonate, lithium carbonate, potassium phosphate, sodium phosphate, cesium phosphate, lithium phosphate, diazabicycloundecene (DBU), diazabicyclononene (DBN), and 1, 3-Tetramethylguanidine (TMG); and combinations thereof.

[60] The method of [57] or [58], wherein the strong base is potassium tert-butoxide, sodium tert-butoxide, lithium tert-butoxide, or LHMDS (lithium hexamethyldisilazide).

[61] The method according to any one of [57] to [60], wherein the reaction is performed in a C1-C10 alcohol solvent alone or in a mixed solvent of a C1-C10 alcohol solvent and an amide-based solvent, an ether-based solvent, an ester-based solvent, an aromatic-based solvent, a halogen-based solvent, a hydrocarbon-based solvent, or a nitrile-based solvent.

[62] A method for producing an oligosaccharide represented by the following formula A-13.

The method comprises the following steps: the compound represented by the following formula A-12 was reacted with DDQ (2, 3-dichloro-5, 6-dicyano-p-benzoquinone) in a mixed solvent of a fluoroalcohol and water to thereby release the 2-naphthylmethyl group from the compound represented by the formula A-12.

[63] The method according to [62], wherein, the fluoroalcohol is selected from the group consisting of hexafluoro-2-propanol (HFIP), 2-Trifluoroethanol (TFE) 2,3, 4, 5-octafluoro-1-pentanol, nonafluoro-tert-butanol, and combinations thereof.

[64] The method according to [62] or [63], which is carried out at-35℃to 70 ℃.

[65] The method according to [62] or [63], which is carried out at-30℃to-10 ℃.

[66] A method for purifying a compound represented by the following formula A-5.

The method comprises the following steps: adding a hydrophobic carrier and water to a water-soluble organic solvent containing the compound represented by the formula a-5 and inclusions, adsorbing the compound represented by the formula a-5 to the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and the water, thereby removing the inclusions, and eluting the compound represented by the formula a-5 from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula a-5.

[67] A method for purifying a compound represented by the following formula A-9.

The method comprises the following steps: adding a hydrophobic carrier and water to a water-soluble organic solvent containing the compound represented by the formula a-9 and inclusions, adsorbing the compound represented by the formula a-9 to the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and the water, thereby removing the inclusions, and eluting the compound represented by the formula a-9 from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula a-9.

[68] A method for purifying a compound represented by the following formula A-12.

The method comprises the following steps: adding a hydrophobic carrier and water to a water-soluble organic solvent containing the compound represented by the formula a-12 and inclusions, adsorbing the compound represented by the formula a-12 to the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and the water, thereby removing the inclusions, and eluting the compound represented by the formula a-12 from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula a-12.

[69] A method for purifying a compound represented by the following formula D-1.

The method comprises the following steps: adding a hydrophobic carrier and water to a water-soluble organic solvent containing the compound represented by the formula D-1 and inclusions, adsorbing the compound represented by the formula D-1 to the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and the water, thereby removing the inclusions, and eluting the compound represented by the formula D-1 from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula D-1.

[70] A method for purifying a compound represented by the following formula D-5.

The method comprises the following steps: adding a hydrophobic carrier and water to a water-soluble organic solvent containing the compound represented by the formula D-5 and inclusions, adsorbing the compound represented by the formula D-5 to the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and the water, thereby removing the inclusions, and eluting the compound represented by the formula D-5 from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula D-5.

[71] A method for purifying a compound represented by the following formula D-8.

(Wherein R ₅ is an aryloxycarbonyl group (COOAr), an acetyl group (Ac), or a2, 2-trichloroethoxycarbonyl group (Troc), and R ₆ is a hydrogen atom, or R ₅ and R ₆ together with the nitrogen atom to which they are bonded form a phthalimide group.)

The method comprises the following steps: adding a hydrophobic carrier and water to a water-soluble organic solvent containing the compound represented by the formula D-8 and inclusions, adsorbing the compound represented by the formula D-8 to the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and the water, thereby removing the inclusions, and eluting the compound represented by the formula D-8 from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula D-8.

[72] A method for purifying a compound represented by the following formula D-10.

The method comprises the following steps: adding a hydrophobic carrier and water to a water-soluble organic solvent containing the compound represented by the formula D-10 and inclusions, adsorbing the compound represented by the formula D-10 to the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and the water, thereby removing the inclusions, and eluting the compound represented by the formula D-10 from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula D-10.

[73] The method according to [69], wherein the inclusion contains a sugar compound other than the compound represented by the formula D-1, and/or a compound derived from a reactant for obtaining the purified compound.

[74] The method according to [70], wherein the inclusion contains a sugar compound other than the compound represented by D-5, and/or a compound derived from a reactant for obtaining the purified compound.

[75] The method according to [71], wherein the inclusion contains a sugar compound other than the compound represented by D-8, and/or a compound derived from a reactant for obtaining the purified compound.

[76] The method according to [72], wherein the inclusion contains a sugar compound other than the compound represented by the formula D-10, and/or a compound derived from a reactant for obtaining the purified compound.

[77] The method according to any one of [69] to [76], wherein the hydrophobic carrier is a resin for reversed-phase partition chromatography packing.

[78] The method of [77], wherein the reverse phase partition chromatography packing resin is selected from the group consisting of poly (styrene/divinylbenzene) polymer gel resins, polystyrene-divinylbenzene resins, polyhydroxymethacrylate resins, styrene-vinylbenzene copolymer resins, polyvinyl alcohol resins, polystyrene resins, polymethacrylate resins, chemically bonded silica gel resins, and combinations thereof.

[79] The method of [78], wherein the chemically bonded silica gel resin is selected from the group consisting of: (1) A resin obtained by reacting a silane coupling agent with silica gel; (2) A resin obtained by chemically bonding dimethyloctadecyl, octadecyl, trimethyloctadecyl, dimethyloctyl, octyl, butyl, ethyl, methyl, phenyl, cyanopropyl, or aminopropyl to silica gel; (3) A resin obtained by chemically bonding a behenyl group or a triacontyl group to silica gel; and (4) combinations of the above (1) to (3).

[80] The method according to [79], wherein the chemically bonded silica gel resin is an octadecyl bonded silica gel resin (ODS resin).

[81] The method according to any one of [69] to [80], wherein the water-soluble organic solvent is a water-soluble alcohol-based solvent, a water-soluble nitrile-based solvent, a water-soluble ether-based solvent, a water-soluble ketone-based solvent, a water-soluble amide-based solvent, a water-soluble sulfoxide-based solvent, or a mixed solvent comprising at least one or more of the above water-soluble organic solvents.

[82] The method according to [81], wherein the water-soluble nitrile solvent is acetonitrile.

[83] The method according to any one of [69] to [82], wherein the organic solvent used in the step of eluting the target from the hydrophobic carrier is a nitrile-based solvent, an ether-based solvent, an ester-based solvent, a ketone-based solvent, a halogen-based solvent, an aromatic-based solvent, or a mixed solvent comprising at least one of the above solvents.

[84] A method for producing a compound represented by the following formula D-8.

The method comprises the following steps: the compound represented by the following formula D-6 is produced (wherein R ₅ is an aryloxycarbonyl group (COOAr), an acetyl group (Ac), or a2, 2-trichloroethoxycarbonyl group (Troc), and R ₆ is a hydrogen atom, or R ₅ and R ₆ form a phthalimide group together with the nitrogen atom to which they are bonded), and then the compound represented by the formula D-6 is formed into a beta-1, 4-glycosidic bond with the compound represented by the following formula D-7, thereby producing the compound represented by the formula D-8.

[85] The method of [84], wherein R ₅ is aryloxycarbonyl (COOAr).

[86] The method of [84] or [85], comprising: the compound represented by the formula D-9 is produced by removing the protecting group of the amino group and the acyl protecting group of the alcohol in the compound represented by the formula D-8 (wherein M ⁺ is sodium ion, lithium ion, potassium ion, or protonated triethylamine cation).

[87] A method for purifying a compound represented by the following formula D-12.

The method comprises the following steps: a step of adding a compound represented by the following formula E-1 (wherein R ₇ is a hydrogen atom, a methyl group, or a methoxy group) to a solution containing the crude compound represented by the formula D-12 to produce a crystalline compound represented by the following formula E-2 (wherein R ₇ is a hydrogen atom, a methyl group, or a methoxy group); and isolating the crystalline compound, followed by a step of extracting the compound represented by the formula D-12 from the isolated crystalline compound.

[88] An oligosaccharide represented by the following formula A-13.

[89] A compound represented by the following formula A-5.

[90] A compound represented by the following formula A-6.

[91] A compound represented by the following formula A-7.

[92] A compound represented by the following formula a-9.

[93] A compound represented by the following formula a-10.

[94] A compound represented by the following formula A-11.

[95] A compound represented by the following formula a-12.

[96] A compound represented by the following formula a-14.

[97] A compound represented by the following formula a-15.

[98] A compound represented by the following formula B-4.

[99] A compound represented by the following formula B-5.

[100] A compound represented by the following formula B-6.

[101] A compound represented by the following formula B-7.

[102] A compound represented by the following formula B-8.

[103] A compound represented by the following formula D-1.

[104] A compound represented by the following formula D-2.

[105] A compound represented by the following formula D-4.

[106] A compound represented by the following formula D-5.

[107] A compound represented by the following formula D-5-FMA.

[108] A compound represented by the following formula D-8.

(Wherein R ₅ is an aryloxycarbonyl group (COOAr), an acetyl group (Ac), or a2, 2-trichloroethoxycarbonyl group (Troc), and R ₆ is a hydrogen atom, or R ₅ and R ₆ form a phthalimide group together with the nitrogen atom to which they are bonded).

[109] A compound represented by the following formula D-8.

[110] A compound represented by the following formula D-9.

(Wherein M ⁺ is a sodium ion, a lithium ion, a potassium ion, or a protonated triethylamine cation).

[111] A compound represented by the following formula D-10.

[112] A compound represented by the following formula D-11.

[113] A crystalline compound represented by the following formula E-2.

(Wherein R ₇ is a hydrogen atom, a methyl group, or a methoxy group).

[114] A compound represented by the following formula D-12, wherein the purity of the compound is 90% or more when measured by HPLC.

[115] The compound of claim 109, wherein the purity is 95% or greater.

[116] An oligosaccharide represented by the following formula D-13.

Effects of the invention

According to the present invention, there can be provided an oligosaccharide represented by the above formula A-13 and a novel process for producing the same, an intermediate for producing the same and a process for producing the same, an oligosaccharide represented by the above formula D-13 and a novel process for producing the same, and an intermediate for producing the same and a process for producing the same.

Drawings

FIG. 1 is a diagram schematically showing an example of a novel process for producing an oligosaccharide represented by the above formula A-13 provided in the present invention.

FIG. 2 is a diagram schematically showing an example of a novel process for producing the compound represented by the above formula A-11 provided in the present invention.

FIG. 3 is a schematic diagram showing an example of a novel process for producing an oligosaccharide represented by the above formula D-13 provided in the present invention.

Detailed Description

Hereinafter, preferred embodiments for carrying out the present invention will be described. The following embodiments are presented as examples of representative embodiments of the present invention, and are not intended to limit the scope of the present invention.

< 1. Process for producing oligosaccharides of formula A-13 >

In one embodiment of the present invention, there are provided novel oligosaccharides represented by formula A-13 and a novel method for producing the same.

In the present invention, the oligosaccharide represented by the formula A-13 means the following oligosaccharide.

The novel synthesis route of the oligosaccharide represented by the above formula A-13 includes the following steps I-1 to I-3.

< Procedure I-1 >)

The step I-1 is a step of producing a compound represented by the following formula A-7, and includes the steps of: the compound represented by the following formula A-5 is produced by reacting a compound represented by the formula A-3 with a compound represented by the following formula A-4 to form an alpha-1, 6-glycosidic bond.

The step I-1 includes the following steps I-1-1 to I-1-3.

< Procedure I-1-1 >)

The step I-1-1 is as follows: the compound represented by formula A-5 is produced by forming an alpha-1, 6-glycosidic bond between the compound represented by formula A-3 and the compound represented by formula A-4. The present step can be carried out by using or applying a known method, but it is preferable that the step is carried out by, for example, the method shown in example 22, for example, adding molecular sieve 4A powder and trimethyltrisulfonate (TMSOTf) in this order to an organic solvent (toluene or the like) and forming an α -1, 6-glycosidic bond between the compound represented by formula a-3 and the compound represented by formula a-4, thereby producing the compound represented by formula a-5. In addition, the compound represented by the formula A-3 and the compound represented by the formula A-4 as starting materials can be produced as follows.

< Preparation of Compound represented by formula A-3 >

In one embodiment of the present invention, the compound represented by formula A-3 can be produced by the following steps, but is not limited to this production method.

First, to a compound represented by the following formula A-1 (3, 4, 6-tri-O-benzyl-1, 2-O- (1-methoxyethylene) - β -D-mannopyranose), for example, water and p-TsOH.H ₂ O are added, and then, they are reacted with triethylamine, thereby producing a compound represented by the following formula A-2.

This step can be preferably performed by the method shown in example 1.

Next, the compound represented by formula A-3 is produced by adding, for example, trichloroacetonitrile and Diazabicycloundecene (DBU) to the compound represented by formula A-2. This step can be preferably performed by the method shown in example 2.

< Preparation of Compound represented by formula A-4 >

In one embodiment of the present invention, the compound represented by the formula A-4 is produced by the following steps X-1 to X-14 or the following steps X-1 to X-8+X-15 to X-16. The following examples show details of the respective steps, which may be carried out by using a general method in the production of monosaccharides or oligosaccharides, or by applying such a general method.

< Procedure X-1 >)

The process X-1 is as follows: the compound represented by the following formula C-2 is produced by protecting the hydroxyl group bonded to the carbon at the 3-position of the compound represented by the following formula C-1 with 2-Naphthylmethyl (NAP).

The compound represented by the formula C-1 as the starting material in this step can be produced by a known method, or a commercially available product can be used. As a commercially available product of the compound represented by the formula C-1, for example, 1, 2:5, 6-di-O-isopropylidene-. Alpha. -D-furanglucose manufactured by Sigma-Aldrich Co. The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 10.

< Procedure X-2 >)

The process X-2 is as follows: the compound represented by the following formula C-3 was produced by subjecting the compound represented by the formula C-2 to acid hydrolysis of two isopropylidene groups and pyranose ring formation.

The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 11.

< Procedure X-3 >)

The process X-3 is as follows: the compound represented by the following formula C-4 was produced by protecting the hydroxyl group on the compound represented by the formula C-3 with an acetyl group.

The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 12.

< Procedure X-4 >)

The process X-4 is as follows: the compound represented by the following formula C-5 is produced by selectively removing only the acetyl group in the acetoxy group to which the carbon at the 1-position of the compound represented by the formula C-4 is bonded.

The steps for producing the compound represented by the formula C-3 to the compound represented by the formula C-5 may be carried out by a one-pot method, for example, as shown in example 12.

< Procedure X-5 >)

The process X-5 is as follows: the compound represented by the following formula C-6 was produced by reacting the compound represented by the formula C-5 with trichloroacetonitrile.

The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 13.

< Procedure X-6 >)

The process X-6 is as follows: the compound represented by the following formula C-8 is produced by reacting a compound represented by the formula C-6 with a compound represented by the following formula C-7.

The compound represented by the formula C-7 can be produced by a known method, or a commercially available product can be used. Examples of the commercial products of the compounds represented by the formula C-7 include 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2-phthalimido-. Beta. -D-glucopyranoside, manufactured by Tokyo chemical Co., ltd. The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 14.

< Procedure X-7 >)

The process X-7 is as follows: a compound represented by the following formula C-9 is produced by detaching an acetyl group from a compound represented by the formula C-8.

The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 15.

In one embodiment of the present invention, the step X-7 is a step of: the acetyl group is cleaved by reacting the compound represented by formula C-8 with a strong base in the presence of trifluoroacetate to produce the compound represented by formula C-9. The elimination reaction of this acetyl group was reported to be carried out using sodium methoxide in methanol (org.biomol. Chem.,2018, 16, 4720-4727), but in this case, an undesired side reaction such as ring opening of phthalimide groups may also occur simultaneously. On the other hand, by using a method in which the reaction is carried out with a strong alkali of an alkoxide system in the presence of an alkyl ester of a perfluorocarboxylic acid, the cleavage of acetyl group can be carried out while suppressing the ring opening of phthalimide group.

The "alkyl ester of perfluorocarboxylic acid" used in the above step is not limited as long as the reaction is allowed to proceed, and is methyl trifluoroacetate, ethyl trifluoroacetate, propyl trifluoroacetate, isopropyl trifluoroacetate, butyl trifluoroacetate, methyl pentafluoropropionate, ethyl pentafluoropropionate, propyl pentafluoropropionate, isopropyl pentafluoropropionate, methyl heptafluorobutyrate, ethyl heptafluorobutyrate, propyl heptafluorobutyrate, isopropyl heptafluorobutyrate, butyl heptafluorobutyrate, methyl nonafluoropentanoate, ethyl nonafluoropentanoate, propyl nonafluoropentanoate, isopropyl nonafluoropentanoate, butyl nonafluoropentanoate, methyl undecanoate, ethyl undecanoate, propyl undecanoate, isopropyl undecanoate, and butyl undecanoate, preferably methyl trifluoroacetate is used.

The "strong base" is not limited as long as the reaction proceeds, and is, for example, selected from the group consisting of: sodium, lithium, and potassium salts of metal amides; sodium, lithium, potassium, cesium, and barium salts of C1-C20 alkoxides; sodium hydride, potassium hydride, lithium hydride, butyllithium, potassium carbonate, sodium carbonate, cesium carbonate, lithium carbonate, potassium phosphate, sodium phosphate, cesium phosphate, lithium phosphate, diazabicycloundecene (DBU), diazabicyclononene (DBN), and 1, 3-Tetramethylguanidine (TMG); examples of the sodium salt, lithium salt, and potassium salt of the C1-C20 alkoxide include lithium methoxide, sodium methoxide, potassium methoxide, lithium ethoxide, sodium ethoxide, potassium ethoxide, lithium isopropoxide, sodium isopropoxide, potassium tert-butoxide, sodium tert-butoxide, potassium tert-butoxide, lithium tert-butoxide, sodium tert-butoxide, or potassium tert-butoxide, and particularly preferably sodium tert-butoxide, lithium tert-butoxide, potassium tert-butoxide, and LHMDS (lithium hexamethyldisilazide).

The solvent used in this step is not limited as long as the reaction is performed, and for example, a C1 to C10 alcohol solvent alone or a mixed solvent of a C1 to C10 alcohol solvent and an amide solvent (dimethylformamide, dimethylacetamide, etc.), an ether solvent (tetrahydrofuran, dimethoxyethane, cyclopentylmethyl ether, etc.), an ester solvent (ethyl acetate, etc.), an aromatic solvent (toluene, etc.), a halogen solvent (methylene chloride, etc.), a hydrocarbon solvent (hexane, etc.), or a nitrile solvent (acetonitrile, etc.) may be used, and methanol or a mixed solvent of methanol and tetrahydrofuran may be preferably used, but is not limited thereto. The "C1 to C10 alcohol solvent" may be replaced by an alcohol having a larger number of carbons, and in view of ease and convenience of obtaining, C1 to C5 alcohols (methanol, ethanol, propanol, butanol, etc.) may be preferably used.

The reaction temperature in the present step is not limited as long as the reaction is carried out, and examples thereof include-20℃to 80℃and preferably 0℃to 70℃and more preferably 20℃to 65℃and particularly preferably 40℃to 60 ℃.

< Procedure X-8 >)

The process X-8 is as follows: the compounds represented by the following formula C-10 were produced by selectively protecting the hydroxyl groups bonded to the carbons at the 4-and 6-positions of D-glucopyranoside in the compounds represented by the formula C-9 using benzaldehyde dimethyl acetal.

The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 16.

< Procedure X-9 >)

The process X-9 is as follows: the compound represented by the following formula C-11 (wherein X ₁ represents a substituent selected from the group consisting of trifluoromethanesulfonyl, nonafluorobutanesulfonyl, 2-nitrobenzenesulfonyl and 4-nitrobenzenesulfonyl) is produced by reacting a compound represented by the formula C-10 with a compound imparting a leaving group selected from the group consisting of trifluoromethanesulfonyloxy, nonafluorobutanesulfonyloxy, 2-nitrobenzenesulfonyloxy and 4-nitrobenzenesulfonyloxy.

This step can be performed by using or applying a known leaving group imparting method, and is preferably performed by the method shown in example 17.

In this step, examples of the "compound for imparting a leaving group selected from the group consisting of trifluoromethanesulfonyl group, nonafluorobutanesulfonyloxy group, 2-nitrobenzenesulfonyloxy group and 4-nitrobenzenesulfonyloxy group" include trifluoromethanesulfonyl anhydride, nonafluoro-1-butanesulfonyl fluoride, bis (nonafluoro-1-butanesulfonic acid) anhydride, 2-nitrobenzenesulfonyl chloride and 4-nitrobenzenesulfonyl chloride, and preferred examples include trifluoromethanesulfonyl anhydride.

The solvent in this step is not limited as long as the reaction is performed, and examples thereof include ethyl acetate, toluene, methylene chloride, acetonitrile, cyclopentyl methyl ether, and t-butyl methyl ether, and preferred examples thereof include ethyl acetate, toluene, and methylene chloride.

The reaction temperature in the present step is not limited as long as the reaction is carried out, and examples thereof include-40℃to 60℃and preferably-30℃to 40℃and more preferably-20℃to 10 ℃.

The present step may be preferably performed in the presence of a base. The base used in the present step is not particularly limited as long as the reaction is performed, and examples thereof include 1-methylimidazole, pyridine, 4-dimethylaminopyridine, picoline, lutidine, and collidine (collidine), and examples thereof include 1-methylimidazole.

< Procedure X-10 >)

The process X-10 is as follows: preparing a compound represented by the following formula C-12 (wherein X ₂ is acetyl) by reacting a compound represented by the formula C-11 with cesium acetate or tetrabutylammonium acetate; or by reacting a compound represented by the formula C-11 with tetrabutylammonium benzoate, to produce a compound represented by the following formula C-12 (wherein X ₂ is benzoyl).

The steric inversion of glucose to mannose has a known conversion reaction, but there is no report on the conversion of glucose-glucosamine disaccharide linked by β -glycosidic bond, in which the protecting group of the hydroxyl group bonded to the 3-carbon of D-glucopyranoside is 2-naphthylmethyl (Nap). By adopting this method, the glucose-mannose steric turnover can be realized, and a mannose-glucosamine disaccharide skeleton linked with a β -glycosidic bond can be constructed with high yield and high selectivity.

The solvent used in the present step is not limited as long as the reaction proceeds, and examples thereof include: dimethyl sulfoxide, dimethylformamide, dimethylacetamide, N-methylpyrrolidone, N-dimethylimidazolidone, sulfolane, tetrahydrofuran or acetonitrile, preferably dimethyl sulfoxide is exemplified.

The reaction temperature in the present step is not limited as long as the reaction is carried out, and examples thereof include 20℃to 80℃and preferably 23℃to 70℃and more preferably 26℃to 60℃and particularly preferably 30℃to 50 ℃.

< Procedure X-11 >)

The process X-11 is as follows: the compound represented by the following formula C-13 was produced by releasing the X ₂ group from the compound represented by the formula C-12 and opening the phthalimide group.

The present step may be carried out by using or applying a known hydrolysis method, and is preferably carried out by the method shown in example 18.

In this step, the compound represented by the formula C-13 thus produced may be used as it is in the next step, or may be isolated and purified by recrystallization. A great advantage of the compound represented by the formula C-13 is that isolation/purification can be performed by crystallization, by which impurities having a similar structure, which are difficult to remove in column purification, can be almost completely removed. In this case, the compound represented by the formula C-13 having a purity of 99% or more by HPLC can be obtained.

The isolation/purification by recrystallization in this step includes, for example: a method of completely removing the solvent from a state dissolved in the solvent by a reduced pressure drying operation; or a method in which tetrahydrofuran is used as a good solvent and isopropyl alcohol is added dropwise as a poor solvent in the presence of a small amount of water.

The recrystallization in this step may be performed using seed crystals of the compound represented by the formula C-13. In the case of using a seed crystal, crystallization can be performed by, for example, the following method: tetrahydrofuran was used as a good solvent, and a part of isopropyl alcohol was added dropwise as a poor solvent in the presence of a small amount of water, and after the addition of seed crystals, crystal precipitation was confirmed, and the remaining isopropyl alcohol was added dropwise.

The step of producing the compound represented by the formula C-13 from the compound represented by the formula C-11 can be performed by, for example, a one-pot method as shown in example 18.

< Procedure X-12 >)

The process X-12 is as follows: the compound represented by the following formula C-14 is produced by ring-closing the ring-opened phthalimido group in the compound represented by the formula C-13 by dehydration condensation.

The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 19.

< Procedure X-13 >)

The process X-13 is as follows: the compound represented by the following formula C-15 was produced by protecting the hydroxyl group bonded to the carbon at the 2-position of D-mannopyranoside in the compound represented by the formula C-14 with a benzyl group.

The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 20.

In one embodiment of the present invention, the step X-13 includes the steps of: the compound represented by the formula C-15 is produced by protecting the hydroxyl group bonded to the carbon 2 of D-mannopyranoside in the compound represented by the formula C-14 with a benzyl group in the presence of lithium t-butoxide or lithium t-amyl alcohol. By performing the step X-15 in the presence of lithium t-butoxide or lithium t-amyl alcohol, the ring opening of phthalimide can be suppressed. In addition, the method can be safely carried out and the scale up is easy compared with the normal conditions using sodium hydride.

The solvent used in the present step is not limited as long as the reaction proceeds, and examples thereof include: dimethylacetamide, dimethylformamide, N-methylpyrrolidone, N-dimethylimidazolidinone, preferably dimethylacetamide.

The reaction temperature in the step is not limited as long as the reaction is carried out, and examples thereof include-20℃to 100℃and preferably-15℃to 70℃and particularly preferably-10℃to 50 ℃.

< Procedure X-14 >)

The process X-14 is as follows: by selectively reducing the benzylidene protecting group in the compound represented by the formula C-15 (for more details, see Angew.chem.int.ed.2005, 44, 1665-1668), a compound represented by the following formula A-4 was produced in which only the hydroxyl group bonded to the 6-carbon of D-mannopyranoside was deprotected.

The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 21.

In this step, the compound represented by the formula A-4 thus produced may be used as it is in the next step, or may be isolated and purified by column purification or the like.

In one embodiment of the present invention, the following steps X-15 and X-16 for performing the stereoscopic inversion of glucose-mannose by the redox reaction are included instead of the steps X-9 to X-12 including the step for performing the stereoscopic inversion of glucose-mannose by the S _N 2 reaction.

< Procedure X-15 >)

The process X-15 is as follows: the compound represented by the following formula C-16 was produced by oxidizing the 2-position of D-glucopyranoside in the compound represented by the formula C-10.

The present process may be performed by using or applying a known method.

< Procedure X-16 >)

The process X-16 is as follows: the compound represented by the following formula C-14 is produced by reducing a ketone group bonded to the carbon at the 2-position of 2-keto-D-glucopyranoside in the compound represented by the formula C-16.

The present process may be performed by using or applying a known method.

The solvent used in the present step is not limited as long as the reaction is performed, and examples thereof include diethyl ether, cyclopentyl methyl ether, t-butyl methyl ether, diisopropyl ether, dipropyl ether, dibutyl ether, and 1, 4-dioxane, and tetrahydrofuran is preferable.

The reaction temperature in this step is not limited as long as the reaction is carried out, and examples thereof include-80℃to 20 ℃. The optimum reaction temperature varies depending on the reducing agent used, as described below.

In one embodiment of the present invention, the oxo group bonded to the 2-carbon of the 2-keto-D-glucopyranoside in the compound represented by formula C-16 is reduced in the presence of a reducing agent selected from the group consisting of lithium tri-sec-butylborohydride, lithium tri-amyl borohydride, lithium diisobutyl-t-butoxyaluminum hydride (LDBBA), the compound represented by formula W below, and combinations thereof.

(Wherein R ₃ is a di-t-butylphenolate or a hydride represented by the following formula, wherein at least two of R ₃ are di-t-butylphenolates.

In this reduction step, for example, when NaBH ₄ is used, the stereoselectivity is low (about 7:3), and it is difficult to efficiently obtain the desired stereospecific inversion of gln→man (org. Biomol. Chem.,2018, 16, 4720-4727). On the other hand, in the case of using the above-listed reducing agent, the selectivity of the stereospecific inversion of Gln.fwdarw.Man was greatly improved (93.6:6.4 to 98.1:1.9) compared with the case of using NaBH ₄.

The compound in which three R ₃ of the compound represented by the formula W are ditert-butyl methylphenolate can be obtained, for example, by adding dibutylhydroxytoluene (885.41 mg,4.02 mmol) to a suspension of lithium aluminum hydride (50.0 mg,1.32 mmol) in tetrahydrofuran (2 mL) at 0℃and stirring at 25 ℃. The compound in which two R ₃ of the compounds represented by the formula W are di-t-butyl methylphenolate can be obtained by the same method using 2 molar equivalents of dibutylhydroxytoluene with respect to 1 molar equivalent of lithium aluminum hydride.

As described above, the reaction temperature in this step is not limited as long as the reaction is carried out, and in the case of using lithium tri-sec-butylborohydride, lithium tripentylborohydride or LDBBA as a reducing agent, the reaction temperature is preferably from-80℃to-20℃and more preferably from-80℃to-30℃and even more preferably from-80℃to-40℃and particularly preferably from-80℃to-50℃and in the case of using the compound represented by formula A as a reducing agent, the reaction temperature is preferably from-20℃to 20℃and more preferably from-15℃to 15℃and particularly preferably from-10℃to 10 ℃. Therefore, in terms of the reaction being carried out at a temperature at which the reaction is easier to handle, the compound represented by formula W is particularly preferable as the reducing agent used in the present step.

< Purification of the Compound shown in A-5 >

In this step I-1-1, the compound represented by the formula A-5 can be obtained in a purified form by the following purification method. The purification method includes: after stopping the reaction between the compound represented by the formula A-4 and the compound represented by the formula A-3, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the produced compound represented by the formula A-5 and inclusions, the compound represented by the formula A-5 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and water, whereby the inclusions are removed, and then the compound represented by the formula A-5 is eluted from the hydrophobic carrier using an organic solvent, whereby the compound represented by the formula A-5 is purified. According to this purification method, in the liquid phase synthesis of oligosaccharide chains, reagent residues remaining after the glycosylation reaction and impurities derived from the sugar donor and the sugar acceptor can be removed easily by performing a washing operation using a small amount of hydrophobic carrier, and reaction inhibition and side reactions due to these impurities can be suppressed, so that high-quality oligosaccharides can be produced in large amounts and efficiently. In addition, the present application can reduce the number of steps of tag desorption and prevent the decrease in the functionality of the tag during oligomerization by utilizing the hydrophobicity of the substrate itself, as compared with the conventionally developed method, and can produce oligosaccharides more efficiently. In particular, in this step, the compound represented by the formula A-5 can be easily separated and purified from the decomposed product derived from the compound represented by the formula A-3 by the above-mentioned purification method.

The purification of the compound represented by the formula A-5 is not limited to the purification in the present step I-1-1. Thus, in one aspect of the invention, there is also provided a method comprising: the method comprises the steps of adding a hydrophobic carrier and water to a water-soluble organic solvent containing a compound represented by the formula A-5 and inclusions, adsorbing the compound represented by the formula A-5 to the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and water to remove the inclusions, and eluting the compound represented by the formula A-5 from the hydrophobic carrier using the organic solvent to thereby purify the compound represented by the formula A-5. Furthermore, the above-described purification method can be applied to purification of organic compounds other than sugar compounds. In addition, in the case where the organic compound to be purified is a sugar compound, a protected oligosaccharide having a sugar chain structure composed of 3 to 15 sugar residues, in which part or all of the hydroxyl groups in the sugar are protected, may be preferably purified, and the protecting group for the sugar chain at this time includes, but is not limited to, alkyl ether, benzyl ether, silyl ether, ester, and carbonate.

The term "inclusion" refers to a compound or reagent other than the protecting oligosaccharide (the compound represented by the formula A-5 in this step), and mainly refers to a reagent used in the synthesis reaction of the protecting oligosaccharide, a residue thereof, a sugar other than the protecting oligosaccharide such as a monosaccharide or disaccharide compound used in the extension reaction of the protecting oligosaccharide, or a by-product produced by the deprotection reaction of the protecting oligosaccharide.

The "hydrophobic carrier" refers to a hydrophobic adsorbent adsorbed to a specific compound containing a sugar compound, and examples thereof include resins for reversed phase partition chromatography packing, and the "resin for reversed phase partition chromatography packing" is selected from the group consisting of poly (styrene/divinylbenzene) polymer gel resins, polystyrene-divinylbenzene resins, polyhydroxymethacrylate resins, styrene-vinylbenzene copolymer resins, polyvinyl alcohol resins, polystyrene resins, polymethacrylate resins, chemically bonded silica gel resins, and combinations thereof, but is not limited thereto.

The "chemically bonded silica gel resin" is preferably selected from the group consisting of (1) a resin obtained by reacting a silane coupling agent with silica gel, (2) a resin obtained by chemically bonding dimethyloctadecyl, octadecyl, trimethyloctadecyl, dimethyloctyl, octyl, butyl, ethyl, methyl, phenyl, cyanopropyl, or aminopropyl to silica gel, (3) a resin obtained by chemically bonding behenyl or triacontyl to silica gel, and (4) a combination of the above (1) to (3), but is not limited thereto.

The "water-soluble organic solvent" is not particularly limited, and a water-soluble alcohol-based solvent (preferably C1 to C4), a water-soluble nitrile-based solvent (acetonitrile, etc.), a water-soluble ether-based solvent (tetrahydrofuran, etc.), a water-soluble ketone-based solvent (acetone, etc.), a water-soluble amide-based solvent (dimethylformamide, etc.), or a water-soluble sulfoxide-based solvent (dimethyl sulfoxide, etc.) may be used, and acetonitrile may be preferably used.

The "organic solvent" used in the step of eluting the target substance from the above-mentioned hydrophobic carrier is not particularly limited, and a nitrile solvent (acetonitrile, etc.), an ether solvent (tetrahydrofuran, etc.), an ester solvent (ethyl acetate, etc.), a ketone solvent (acetone, etc.), a halogen solvent (methylene chloride, etc.), an aromatic solvent (toluene, etc.), or a mixed solvent containing at least one or more of the above-mentioned solvents may be used, and acetonitrile, ethyl acetate, tetrahydrofuran, toluene may be preferably used.

The above-mentioned purification step is not particularly limited, and may be carried out at a temperature of 0℃to 50 ℃.

The compound represented by the formula A-7 can be produced from the compound represented by the formula A-5 by the following steps I-1-2 to I-1-3 after the step I-1-1, but is not limited to these production steps.

< Procedure I-1-2 >)

The step I-1-2 is as follows: deprotection of the 4-methoxyphenyl group from the compound represented by the above formula A-5 gives a compound represented by the following formula A-6.

In one embodiment of the present invention, the present process is a process comprising: deprotection of 4-methoxyphenyl by reacting a compound represented by the above formula A-5 with λ3-iodide in fluoroalcohol and water to produce a compound represented by the above formula A-6. This step can be preferably performed by the method shown in example 23, for example.

The above "λ3-iodide" means a trivalent super-valent iodine compound. In one embodiment, is a compound of formula R ⁴－I(OR⁵)₂ (wherein R ⁴ is unsubstituted or substituted phenyl, R ⁵ is selected from the group consisting of H, acetoxy, trifluoroacetoxy, tosyloxy, methanesulfonyloxy, and combinations thereof). As shown in the definition of the above formula, R ⁴ may be a "substituted phenyl group", and examples of the substituent include a linear or branched saturated or unsaturated hydrocarbon group, an oxygen-containing group (an alkoxy group, an ester, etc.), a nitrogen-containing group (a cyano group, an azide group, etc.), a halogen atom (for example, a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, etc.), and more preferably a hydrocarbon group, an oxygen-containing substituent, a halogen atom, etc. In the case where these substituents contain carbon, for example, a substituent having 1 to 5 carbons or a substituent having 1 to 3 carbons can be preferably used. Specific examples of λ3-iodide include: [ bis (trifluoroacetoxy) iodo ] benzene (PIFA), [ hydroxy (tosyloxy) iodo ] benzene (HTIB), (diacetoxy iodo) benzene (PIDA), [ bis (trifluoroacetoxy) iodo ] pentafluorobenzene, and [ hydroxy (methanesulfonyloxy) iodo ] benzene, but is not limited thereto.

The "fluoroalcohol" used in the above step refers to a fluorine-containing alcohol compound having fluorine in all carbons except the carbon bonded to the alcohol. The fluoroalcohol preferably has more fluorine as long as fluorine substitution is allowed. The fluoroalcohol includes a fluoroaliphatic alcohol, but is not limited thereto. The hydrocarbon moiety in the fluoroaliphatic alcohol may be saturated or unsaturated, may be linear or branched, or may be cyclic. The fluoroaliphatic alcohol is, for example, a fluoroaliphatic alcohol ₂～C₈, preferably a fluoroaliphatic alcohol ₂～C₅, and more preferably a fluoroaliphatic alcohol ₂～C₃. As a specific example of the fluoroalcohol, a fluorine alcohol, examples thereof include hexafluoro-2-propanol (HFIP), 2-Trifluoroethanol (TFE) 2,3, 4, 5-octafluoro-1-pentanol, nonafluoro-tert-butanol, combinations thereof, however, the present invention is not limited thereto.

The present step is carried out in the presence of the above fluoroalcohol and "water". The amount of water may be appropriately set from the viewpoint of achieving a high yield of the product, etc., and for example, may be about 1.0 equivalent or more, about 1.5 equivalent or more, about 2.0 equivalent or more, or about 2.5 equivalent or more in terms of a molar ratio with respect to the compound represented by formula a-5, and further may be about 10 or less, about 8 or less, about 5 or less, or about 3 or less in terms of a volume ratio with respect to the compound represented by formula a-5.

In this step, an "additive" may be further added to the fluoroalcohol and water. The additive is preferably selected from the group consisting of sodium dihydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, trifluoroacetic acid, and combinations thereof. The amount of the additive may be appropriately set, and for example, may be about 0.5 to 8 equivalents, about 1 to 6 equivalents, or about 1.5 to 5 equivalents with respect to the compound represented by formula A-5.

< Procedure I-1-3 >)

The step I-1-3 is a step of producing the compound represented by the formula A-7 from the compound represented by the formula A-6. The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 24.

In one embodiment of the present invention, the present process is a process comprising: the compound represented by the above formula A-6 is reacted with 2, 2-trifluoro-N-phenyliminoacetyl chloride (TFPC) in the presence of DBU to produce the compound represented by the above formula A-7. By using the DBU as the base to be used, for example, the equivalent weight of TFPC can be reduced as compared with the case of using potassium carbonate, and the target product can be obtained in high yield. Since TFPC is an expensive reagent, the yield improvement of this process is very beneficial in commercial production.

The solvent used in the present step is not limited as long as the reaction is performed, and examples thereof include methylene chloride, toluene, ethyl acetate, acetonitrile, and tetrahydrofuran, and preferred examples thereof include methylene chloride.

The reaction temperature in the present step is not limited as long as the reaction is carried out, and it is preferably from-20℃to 40℃and more preferably from-10℃to 35℃and particularly preferably from 0℃to 30 ℃.

The present step is preferably carried out in the presence of a dehydrating agent. The dehydrating agent in this step is not limited as long as the reaction is performed, and examples thereof include molecular sieves, preferably molecular sieve 4A powder having a powder particle diameter of 10 μm or less.

In this step, the compound represented by A-7 produced may be used as it is in the next step, or may be isolated and purified by column purification or the like, as long as the removal of the base used in the reaction is performed. As the isolation/purification using a chromatographic column, for example, isolation/purification using silica gel as a stationary phase and methylene chloride or toluene-ethyl acetate mixed solvent system as a mobile phase can be cited.

< Procedure I-2 >

The step I-2 is a step of producing a compound represented by the following formula A-10, and includes the steps of: the compound represented by the following formula A-9 is produced by forming a beta-1, 4-glycosidic bond between the compound represented by the above formula A-7 and the compound represented by the following formula A-8.

The step I-2 includes the following steps I-2-1 to I-2-2.

< Procedure I-2-1 >)

The step I-2-1 is as follows: the compound represented by formula A-9 is produced by reacting the compound represented by formula A-7 with the compound represented by formula A-8 to form a beta-1, 4-glycosidic bond. The compound represented by the formula A-8 can be produced by a known method, or a commercially available product can be used. Examples of the commercial products of the compounds represented by the formula A-8 include 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2-phthalimido-. Beta. -D-glucopyranoside, manufactured by Tokyo chemical Co. The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 25.

< Purification of Compound represented by formula A-9 >

In this step I-2-1, the compound represented by the formula A-9 can be obtained in a purified form by the following purification method. The purification method includes: after stopping the reaction between the compound represented by the formula A-7 and the compound represented by the formula A-8, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the produced compound represented by the formula A-9 and impurities, the compound represented by the formula A-9 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and water, whereby the impurities are removed, and then the compound represented by the formula A-9 is eluted from the hydrophobic carrier using an organic solvent, whereby the compound represented by the formula A-9 is purified. As described in the above-mentioned method for purifying a compound represented by the formula A-5, the method for purifying a compound can produce a large amount of high-quality oligosaccharides efficiently in liquid phase synthesis of oligosaccharide chains by using a small amount of a hydrophobic carrier. In particular, the compound represented by the formula A-8 as a monosaccharide and the compound represented by the formula A-9 as a tetrasaccharide have extremely close polarities in normal phase silica gel column chromatography, and for example, have the same Rf value under the condition of a typical column solvent system of hexane-ethyl acetate, and separation is difficult, whereas by using the purification method of the present invention, monosaccharides and tetrasaccharides having extremely close polarities can be easily separated.

The purification of the compound represented by the formula A-9 is not limited to the purification in the present step I-2-1. Accordingly, in one aspect of the present invention, there is also provided a method comprising: the method comprises the steps of adding a hydrophobic carrier and water to a water-soluble organic solvent containing a compound represented by the formula A-9 and inclusions, adsorbing the compound represented by the formula A-9 to the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and water to remove the inclusions, and eluting the compound represented by the formula A-9 from the hydrophobic carrier using the organic solvent to thereby purify the compound represented by the formula A-9.

The term "inclusion" refers to a compound or reagent other than the protecting oligosaccharide (the compound represented by the formula A-9 in this step), and mainly refers to a reagent used in the synthesis reaction of the protecting oligosaccharide, a residue thereof, a sugar other than the protecting oligosaccharide such as a monosaccharide or disaccharide compound used in the extension reaction of the protecting oligosaccharide, or a by-product produced by the deprotection reaction of the protecting oligosaccharide. The "hydrophobic carrier" (resin for filling in reverse phase partition chromatography, etc.), "water-soluble organic solvent", "organic solvent", and the purification temperatures used in this step are the same as those described in the method for purifying the compound represented by the above formula A-5.

< Procedure I-2-2 >)

The step I-2-2 is a step of producing a compound represented by the formula A-10 from a compound represented by the formula A-9.

In one embodiment of the present invention, the present step I-2-2 is a step of: the compound represented by the formula A-10 is preferably produced by, for example, the method shown in example 26 by reacting a compound represented by the formula A-9 with a strong base in the presence of an alkyl ester of a perfluorocarboxylic acid to cause the acetyl group to be detached (deacetylation reaction) in a solvent. The deacetylation reaction can be carried out in the same manner as the deacetylation reaction described in the step X-7, except that the substrate is different, and the deacetylation reaction can be carried out while suppressing the ring opening of the phthalimide group by using a method in which the substrate is reacted with a strong base in the presence of an alkyl ester of a perfluorocarboxylic acid.

The deacetylation reaction is not limited to the one used in the present step I-2-2. Accordingly, in one embodiment of the present invention, there is also provided a method for producing the compound represented by the above formula A-10, which comprises a step of reacting the compound represented by the above formula A-9 with a strong base in the presence of an alkyl ester of a perfluorocarboxylic acid.

The alkyl ester of perfluorocarboxylic acid, strong base, solvent, and reaction temperature in this step are as described in the above step X-7.

< Procedure I-3 >

The step I-3 is a step of producing an oligosaccharide represented by the above formula A-13, and includes the steps of: the compound represented by the following formula A-12 is produced by forming a beta-1, 2-glycosidic bond between the compound represented by the above formula A-10 and the compound represented by the following formula A-11.

In one embodiment of the present invention, the step I-3 includes the following steps I-3-1 to I-3-2.

< Procedure I-3-1 >)

The step I-3-1 is as follows: the compound represented by the above formula A-12 is produced by forming a beta-1, 2-glycosidic bond between the compound represented by the above formula A-10 and the compound represented by the above formula A-11. The glycoside bonding step may be performed by using or applying a known method, and is preferably performed by the method shown in example 27. The compound represented by the formula A-11 can be produced as follows. The compound represented by the formula A-12 may be purified as described below.

< Preparation of Compound represented by formula A-11 >

In one embodiment, the compound represented by the formula A-11 is produced by a process comprising a process Y-1 and a process Y-2.

The (step Y-2) is as follows: a compound represented by the following formula B-5 is produced by adding lithium t-butoxide or lithium t-amyl alcohol to a solvent comprising a compound represented by the formula B-4 and a benzyl halide or benzyl sulfonate, and protecting the hydroxyl group present in the compound represented by the formula B-4 with a benzyl group.

The process Y-1 includes a process Y-1-1 and a process Y-1-2, and the process Y-2 includes a process Y-2-1 to a process Y-2-3.

< Procedure Y-1-1 >)

The step Y-1-1 is as follows: the compound represented by the above formula B-3 is produced by forming a beta-1, 4-glycosidic bond between the compound represented by the above formula B-1 and the compound represented by the above formula B-2. As a commercially available product of the compound represented by the formula B-1, 2,3,4, 6-tetra-O-acetyl-alpha-D-galactopyranosyl 2, 2-trichloroiminoacetate (86520-63-0) is exemplified by Tokyo chemical Co. Further, examples of the commercial products of the compound represented by the formula B-2 include 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2-phthalimido-. Beta. -D-glucopyranoside, manufactured by Tokyo chemical Co., ltd. The present step can be carried out by using or applying a known method, and preferably can be carried out by, for example, the method shown in example 3, and for example, the compound shown in the following formula B-3 can be produced by sequentially adding the solution containing the compound shown in the formula B-1, the molecular sieve 4A powder, and the trimethylsilyl triflate (TMSOTF) to the solution containing the compound shown in the formula B-2.

< Procedure Y-1-2 >)

The step Y-1-2 is as follows: the acetyl group is released from the compound represented by B-3 to produce a compound represented by the following formula B-4.

The deprotection of the acetyl group (AcO) is carried out by a known method, preferably, for example, by a method shown in example 4, for example, by reacting a compound represented by the formula B-3 with a strong base in the presence of trifluoroacetate in a solvent such as toluene, and thereby removing the acetyl group, whereby a compound represented by the formula B-4 can be produced.

< Procedure Y-2-1 >)

The process Y-2-1 is as follows: the compound represented by the above formula B-5 is produced by protecting a plurality of hydroxyl groups present in the compound represented by the above formula B-4 with benzyl groups.

In one embodiment, the present step Y-2-1 is the following step: the compound represented by the above formula B-5 is produced by adding lithium tert-butoxide or lithium tert-amyl alcohol to a solvent comprising the compound represented by the above formula B-4 and benzyl halide (benzyl bromide, benzyl chloride, benzyl fluoride, or benzyl iodide) or benzyl sulfonate, and protecting a plurality of hydroxyl groups present in the compound represented by the above formula B-4 with benzyl groups. When it is necessary to simultaneously benzyl a plurality of hydroxyl groups in a phthalimide group-protected sugar derivative such as the compound N-2) shown in the above B-4, it is necessary to carry out the reaction while suppressing the ring opening of the phthalimide group. But has the following disadvantages: in the case of the conventional NaH/DMAc used in the benzylation reaction, the yield greatly varies depending on the amount of sodium hydroxide in NaH, and NaH/DMAc is not easily used in a large amount of synthesis methods due to the danger of mixing and explosion hazards. The present inventors have found the following methods: by performing the above-mentioned benzylation reaction, a plurality of hydroxyl groups can be simultaneously benzylated under milder conditions while suppressing the ring opening of the phthalimide group.

The above-mentioned benzylation reaction is not limited to the one used in the present step Y-2-1. Accordingly, in one embodiment of the present invention, there is also provided a method for producing the compound represented by the above formula B-5, comprising the steps of: to a solution containing the compound represented by the above formula B-4 and a solvent containing benzyl halide (benzyl bromide, benzyl chloride, benzyl fluoride, or benzyl iodide) or benzyl sulfonate, lithium tert-butoxide or lithium tert-amyl alcohol is added, and the hydroxyl group present in the compound represented by the above formula B-4 is protected with benzyl.

The solvent used in the present step is not particularly limited as long as the reaction proceeds, and may be used: an amide solvent (dimethylformamide, dimethylacetamide, etc.), an ether solvent (tetrahydrofuran, dimethoxyethane, etc.), an aromatic solvent (toluene, etc.), a hydrocarbon solvent (hexane, etc.), a urea solvent, or a mixed solvent containing at least one of the above solvents, more preferably an amide solvent (dimethylformamide, dimethylacetamide, etc.) can be used.

The reaction in this step is preferably carried out at 0 to 60 ℃, more preferably at 30 to 50 ℃.

< Purification of Compound represented by formula B-5 >

The compound represented by the above formula B-5 can be purified by the following steps. In this step, the compound represented by the formula B-5 is prepared by opening the phthalimide group in the compound, then forming a salt with cinchonidine to give a crystalline compound represented by the following formula B-6 as a cinchonidine salt, separating the crystalline compound represented by the formula B-6 from the amorphous substance, removing cinchonidine from the compound represented by the formula B-6 by adding a solvent to give a compound represented by the following formula B-7, and then closing the ring of the ring-opened phthalimide group in the compound represented by the formula B-7, thereby regenerating the compound represented by the formula B-5.

Since the compound represented by the formula B-6 (cinchonidine salt of the compound represented by the formula B-7) is crystalline and the compounds represented by the formulae B-3, B-4 and B-5 are compounds which do not crystallize, the phthalimide in the compound represented by the formula B-5 is once ring-opened, the carboxylic acid moiety in the phthalimide group thus formed forms a salt with cinchonidine, the compound represented by the formula B-5 can be crystallized, the crystalline substance is separated from the amorphous substance, and then, for example, an acidic aqueous solution and a solvent are added to remove cinchonidine in the compound represented by the formula B-6, and then, the phthalimide is ring-closed again, whereby the highly purified compound represented by the formula B-5 can be obtained. The ring opening and ring closing of the phthalimide may be performed by a known method, and the ring opening of the phthalimide may be performed by adding sodium hydroxide to methanol-tetrahydrofuran, and the ring closing of the phthalimide may be performed by adding Carbonyl Diimidazole (CDI) to a tetrahydrofuran solvent, for example. This step can be preferably performed by the methods shown in examples 6 and 7, for example.

< Procedure Y-2-2 >)

The process Y-2-2 is as follows: by detaching 4-methoxyphenyl from the compound represented by the formula B-5, a compound represented by the following formula B-8 is produced.

In one embodiment, the present step Y-2-2 is the following step: the compound represented by the formula B-5 is preferably produced by, for example, the method shown in example 8 by reacting a compound represented by the formula B-5 with λ3-iodide in fluoroalcohol and water to disengage 4-methoxyphenyl group. This step can be performed in accordance with the above-described step I-1-2, and the same compound as that used in the above-described step I-1-2 can be used for the fluoroalcohol and λ3-iodide used in this step.

< Procedure Y-2-3 >)

The step Y-2-3 is a step of producing the compound represented by the formula A-11 from the compound represented by the formula B-8.

In one embodiment of the present invention, the present step Y-2-3 is a step of: the reaction of a compound represented by the above formula B-8 with 2, 2-trifluoro-N-phenyliminoacetyl chloride (TFPC) in the presence of N-methylimidazole to give a compound represented by the above formula A-11 is preferably carried out by the method shown in example 9. As described in the same reaction in step I-1-3, the use of N-methylimidazole as the base used can reduce the equivalent weight of TFPC as compared with the case of potassium carbonate, for example, and can yield the target product in high yield. The solvent used, the reaction temperature, and preferably the presence of a dehydrating agent, and isolation/purification by column purification or the like are also similar to the reaction in the above-mentioned steps I-1 to 3.

< Purification of the Compound shown in A-12 >

In the above step I-3-1, the compound represented by the formula A-12 can be obtained in a purified form by the following purification method. The purification method includes: after stopping the reaction between the compound represented by the formula A-10 and the compound represented by the formula A-11, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the produced compound represented by the formula A-12 and inclusions, the compound represented by the formula A-12 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and water, whereby the inclusions are removed, and then the compound represented by the formula A-12 is eluted from the hydrophobic carrier using an organic solvent, whereby the compound represented by the formula A-12 is purified. As described in the above-mentioned method for purifying a compound represented by the formula A-5, according to the above-mentioned purification method, a large amount of high-quality oligosaccharides can be efficiently produced in the liquid phase synthesis of oligosaccharide chains by using a small amount of a hydrophobic carrier.

The purification of the compound represented by the formula A-12 is not limited to the purification in the present step I-3-1. Accordingly, in one aspect of the present invention, there is also provided a method comprising: the method comprises the steps of adding a hydrophobic carrier and water to a water-soluble organic solvent containing a compound represented by the formula A-12 and inclusions, adsorbing the compound represented by the formula A-12 to the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and water to remove the inclusions, and eluting the compound represented by the formula A-12 from the hydrophobic carrier using the organic solvent to thereby purify the compound represented by the formula A-12.

The term "inclusion" refers to a compound or reagent other than the protecting oligosaccharide (the compound represented by the formula A-12 in this step), and mainly refers to a reagent used in the synthesis reaction of the protecting oligosaccharide, a residue thereof, a sugar other than the protecting oligosaccharide such as a monosaccharide or disaccharide compound used in the extension reaction of the protecting oligosaccharide, or a by-product produced by the deprotection reaction of the protecting oligosaccharide.

The "hydrophobic carrier" (resin for reversed phase partition chromatography packing, etc.), "water-soluble organic solvent", "organic solvent", and the purification temperatures used in the above steps are the same as those described in the method for purifying the compound represented by the above formula A-5.

< Procedure I-3-2 >

The step I-3-2 is a step of producing an oligosaccharide represented by the formula A-13 from the compound represented by the formula A-12.

In one embodiment of the present invention, the present step I-3-2 is a step of: the oligosaccharide represented by the above formula A-13 is preferably produced by, for example, the method shown in example 28-1 by reacting a compound represented by the above formula A-12 with DDQ (2, 3-dichloro-5, 6-dicyano-p-benzoquinone) in a mixed solvent of a fluoroalcohol and water to thereby release the 2-naphthylmethyl group in the compound represented by the above formula A-12 (a 2-naphthylmethylation reaction).

Regarding the above-mentioned demethylation reaction of 2-naphthyl, the present inventors have found that by reacting (acting) 2, 3-dichloro-5, 6-dicyano-p-benzoquinone with a substrate to which 2-naphthylmethyl groups are bonded via oxygen atoms in a fluoroalcohol and water, the reaction can be carried out under mild conditions with good stirring properties, and the demethylated product can be obtained in high yield. The advantages of this de-2-naphthylmethylation reaction will be described in more detail below. In the above-described 2-naphthylmethyl-removing reaction of the present invention, a 2-naphthylmethyl-removed product can be obtained in high yield under mild conditions from a substrate such as a sugar to which a 2-naphthylmethyl group is bonded via an oxygen atom. The inventors have found that the reaction can be performed with good reproducibility without deteriorating the stirring property and the adhesion to the wall surface of the vessel due to the 2, 3-dichloro-5, 6-dicyano-p-benzoyl hydroquinone as a by-product, and that the method is suitable for mass synthesis of the above-mentioned products. In addition, since the abnormal freezing point of HFIP-H ₂ O was lowered, solidification of the solvent was not confirmed even if the reaction temperature was lowered to-30℃and a wide temperature range (melting point HFIP: -3.3℃and H ₂ O:0 ℃) was applicable depending on the reactivity of the reaction substrate. In addition, it was found that when the decalin methylation reaction of the compound represented by the formula A-12 having many benzyl groups was performed, DDQ was used as an oxidizing agent and HFIP-H ₂ O was used as a solvent, whereby the reaction was performed with superior selectivity as compared with the conventional conditions. For the conversion reaction, in a large number of reported examples, methylene chloride-water double-layer reaction conditions were applied, in which case the debenzylation of a plurality of benzyl groups was carried out at a constant rate, and the yield remained at a moderate level. Although there are reported examples of improvement conditions using β -pinene as an additive (refer to j. Org. Chem.,2017, 82, 3926, etc.), the yield stays at a moderate level for compounds having multiple Bn groups (refer to angel. Chem. Int. Ed.2021, 60, 19287, etc.). further, the selectivity of the substrate having 10 or more benzyl groups, particularly for the compound represented by the formula A-12, is not sufficiently known. In addition, in the methylene chloride-water system, there is a problem that DDQ and a byproduct derived from DDQ cause deterioration of stirring property, and it cannot be said that the reaction conditions suitable for mass synthesis are suitable. On the other hand, in the DDQ/HFIP-H ₂ O system of the present method, a high selectivity of 85% or more is achieved in the compound represented by the formula A-12 having a benzyl group at 15. the method did not confirm the deterioration of the stirring property due to DDQ as described above. Furthermore, since the abnormal freezing point of HFIP-H ₂ O was lowered, solidification of the solvent was not confirmed even if the reaction temperature was lowered to-30 ℃, and a wide temperature range (melting point HFIP: -3.3 ℃ C., H ₂ O:0 ℃ C.) was applicable depending on the reactivity of the reaction substrate.

The aforementioned 2-naphthylene-removed methylation reaction is not limited to the reaction in the present step I-3-1. Accordingly, in one embodiment of the present invention, there is also provided a method for producing an oligosaccharide represented by the above formula A-13, comprising the steps of: the compound represented by the formula A-12 is reacted with DDQ (2, 3-dichloro-5, 6-dicyano-p-benzoquinone) in a mixed solvent of a fluoroalcohol and water to disengage the 2-naphthylmethyl group in the compound represented by the formula A-12.

The "fluoroalcohol" is not limited as long as the reaction proceeds, preferably selected from the group consisting of hexafluoro-2-propanol (HFIP), 2-Trifluoroethanol (TFE) 2,3, 4, 5-octafluoro-1-pentanol, nonafluoro-tert-butanol, and combinations thereof.

The aforementioned 2-naphthylene-removing methylation reaction is not limited as long as the reaction is carried out, and is preferably carried out at-35℃to 70℃and more preferably at-30℃to-10 ℃.

According to the above production method, an oligosaccharide represented by the following formula A-13 is provided.

The oligosaccharide represented by the formula A-13 may contain a modification thereof, for example, a modification containing a protecting group such as chlorobenzyl instead of benzyl in the oligosaccharide represented by the formula A-13, as long as the oligosaccharide has the same function or action as the oligosaccharide.

< Purification of Compound represented by formula A-13 >

The compound represented by the above formula A-13 can be purified by the following steps. In this step, the phthalimide group in the compound represented by the formula A-13 is ring-opened, then, a salt is formed with (R) - (+) -1- (1-naphthyl) ethylamine to produce a crystalline compound represented by the following formula A-14, the crystalline compound represented by the formula A-14 is separated from the amorphous substance, and then, an acidic aqueous solution and a solvent are added to remove (R) - (+) -1- (1-naphthyl) ethylamine in the compound represented by the formula A-14 to produce a compound represented by the following formula A-15, and then, the ring-opened phthalimide group in the compound represented by the formula A-15 is ring-closed, whereby the compound represented by the formula A-13 can be regenerated.

The compound represented by the formula A-14 (the (R) - (+) -1- (1-naphthyl) ethylamine salt of the compound represented by the formula A-15) is crystalline, and the compound represented by the formula A-13 is a compound which does not crystallize, and therefore, the phthalimide in the compound represented by the formula A-13 is once ring-opened to form a salt between the carboxylic acid moiety in the phthalimide group thus formed and the (R) - (+) -1- (1-naphthyl) ethylamine, and the obtained crystalline substance is separated from the amorphous substance, and then, an acidic aqueous solution and a solvent are added to remove the (R) - (+) -1- (1-naphthyl) ethylamine in the compound represented by the formula A-14, thereby obtaining the compound represented by the formula A-15, and then, the phthalimide is ring-closed again, whereby the compound represented by the formula A-13 can be obtained with high purity. The ring opening and ring closing of the phthalimide may be performed by a known method, and the ring opening of the phthalimide may be performed by adding sodium hydroxide to methanol-tetrahydrofuran, and the ring closing of the phthalimide may be performed by adding Carbonyl Diimidazole (CDI) to a tetrahydrofuran solvent, for example. This step can be preferably performed by the method shown in example 28-2.

< 2. Process for producing oligosaccharides of formula D-13 >

In one embodiment of the present invention, there are provided novel oligosaccharides represented by formula D-13 and a novel method for producing the same. In the present invention, the oligosaccharide represented by the formula D-13 means the following oligosaccharide.

The novel synthesis route of the oligosaccharide represented by the formula D-13 includes the following steps II-1 to II-4.

< Procedure II-1 >)

The step I-1 is a step of producing a compound represented by the following formula D-2, and includes the steps of: the oligosaccharide represented by the following formula A-13 and the compound represented by the following formula A-3 are formed into an alpha-1, 3-glycosidic bond to produce a compound represented by the following formula D-1.

In one embodiment of the present invention, the step II-1 includes the following steps II-1-1 to II-1-2.

< Procedure II-1-1 >)

The step II-1-1 is as follows: the compound represented by the formula D-1 is produced by forming an alpha-1, 3-glycosidic bond between the compound represented by the formula A-13 and the compound represented by the formula A-3. The present step can be preferably performed by using or applying a known method, and for example, the method shown in example 52 can be performed by adding molecular sieve 4A powder and trimethylsilicone triflate (TMSOTf) in order to an organic solvent (toluene, etc.), and forming an α -1, 3-glycosidic bond between the compound represented by formula a-13 and the compound represented by formula a-3, thereby producing the compound represented by formula D-1.

< Purification of Compound represented by formula D-1 >

In this step II-1-1, the compound represented by the formula D-1 can be obtained in a purified form by the following purification method. The purification method includes: after stopping the reaction between the compound represented by the formula A-13 and the compound represented by the formula A-3, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the produced compound represented by the formula D-1 and inclusions, the compound represented by the formula D-1 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and water, whereby the inclusions are removed, and then the compound represented by the formula D-1 is eluted from the hydrophobic carrier using an organic solvent, whereby the compound represented by the formula D-1 is purified. As described in the above-mentioned method for purifying the compound represented by the formula A-5 in the step I-1-1, the use of a small amount of the hydrophobic carrier in the purification method enables the production of high-quality oligosaccharides in a large amount and efficiently in the liquid-phase synthesis of oligosaccharide chains.

The purification of the compound represented by the formula D-1 is not limited to the purification in this step. Thus, in one aspect of the invention, there is also provided a method comprising: the method comprises the steps of adding a hydrophobic carrier and water to a water-soluble organic solvent containing a compound represented by the formula D-1 and inclusions, adsorbing the compound represented by the formula D-1 on the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and water to remove the inclusions, and eluting the compound represented by the formula D-1 from the hydrophobic carrier using the organic solvent to thereby purify the compound represented by the formula D-1.

The term "inclusion" refers to a compound or reagent other than the protecting oligosaccharide (compound represented by the formula D-1 in this step), and mainly refers to a reagent used in the synthesis reaction of the protecting oligosaccharide, a residue thereof, a sugar other than the protecting oligosaccharide such as a monosaccharide or disaccharide compound used in the extension reaction of the protecting oligosaccharide, or a by-product produced by the deprotection reaction of the protecting oligosaccharide. The "hydrophobic carrier" (resin for filling by reversed phase partition chromatography, etc.), "water-soluble organic solvent", "organic solvent", and the purification used in this step were the same as those described in the method for purifying the compound represented by formula A-5 in step I-1-1.

< Procedure II-1-2 >)

The step II-1-2 is as follows: the compound represented by the formula D-2 is produced by detaching an acetyl group from the compound represented by the formula D-1. The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 53.

In one embodiment of the present invention, the step II-1-2 is as follows: the compound represented by the formula D-2 is produced by reacting the compound represented by the formula D-1 with a strong base in the presence of an alkyl ester of a perfluorocarboxylic acid to thereby release an acetyl group. The deacetylation reaction may be carried out in the same manner as the deacetylation reaction described in the above step X-7, except that the substrate is different, and according to this method, the deacetylation reaction can be carried out while suppressing the ring opening of the phthalimide group by using a method of reacting with a strong base in the presence of an alkyl ester of a perfluorocarboxylic acid.

The deacetylation reaction is not limited to the one used in the present step II-1-2. Accordingly, in one embodiment of the present invention, there is also provided a method for producing the compound represented by the above formula D-2, which comprises a step of reacting the compound represented by the above formula D-1 with a strong base in the presence of an alkyl ester of a perfluorocarboxylic acid.

< Procedure II-2 >

The step II-2 is as follows: after the compound represented by the following formula D-5 is produced, the amino group in the compound represented by the above formula D-5 is protected with a protecting group selected from the group consisting of an aryloxycarbonyl group (COOAr), an acetyl group (Ac), a2, 2-trichloroethoxycarbonyl group (Troc), and a phthalimide group (Pht), to produce a compound represented by the following formula D-6 (wherein R ₅ is an aryloxycarbonyl group (COOAr), an acetyl group (Ac), or a2, 2-trichloroethoxycarbonyl group (Troc), R ₆ is a hydrogen atom, or R ₅ and R ₆ form a phthalimide group together with the nitrogen atom to which they are bonded); or removing acetyl (Ac) from the compound represented by the following formula D-4 to produce a compound represented by the following formula D-6 (wherein R ₅ and R ₆ together with the nitrogen atom to which they are bonded form a phthalimide group), and the step II-2 comprises a step of forming a beta-1, 2-glycosidic bond between the compound represented by the above formula D-2 and the compound represented by the following formula D-3 to produce the compound represented by the following formula D-4.

In one embodiment of the present invention, the step II-2 includes the following steps II-2-1 to II-2-3.

< Procedure II-2-1 >

The step II-2-1 is as follows: the compound represented by the above formula D-4 is produced by forming a beta-1, 2-glycosidic bond between the compound represented by the above formula D-2 and the compound represented by the above formula D-3. The glycoside bonding step may be performed by using or applying a known method, and is preferably performed by a method shown in example 54, for example, by sequentially adding molecular sieve 4A powder and trimethylsilyl triflate (TMSOTf) to an organic solvent (toluene, etc.), and forming β -1, 2-glycosidic bond between the compound represented by formula D-2 and the compound represented by formula D-3, thereby producing the compound represented by formula D-4.

< Preparation of Compound represented by formula D-3 >

The compounds represented by the above formula D-3 can be produced in the following small steps Z-1 to Z-3.

)

First, the hydroxyl group on the compound represented by the formula A-8, which is also used in the above-mentioned step Y-1, is protected with an acetyl group to produce a compound represented by the following formula F-1.

The step may be performed by using or applying a known method, and preferably may be performed by, for example, the method shown in example 34, and may be performed by adding triethylamine, dimethylaminopyridine and acetic anhydride to an ethyl acetate solution of the compound represented by formula a-8, but is not limited to this method.

)

Then, the 4-methoxyphenyl group is separated from the compound represented by the formula F-1 to produce a compound represented by the following formula F-2.

In one embodiment of the present invention, the small step Z-2 is the following step: the compound represented by the formula F-2 is produced by reacting a compound represented by the formula F-1 with λ3-iodide in fluoroalcohol and water to release 4-methoxyphenyl, and is preferably produced by the method shown in example 35. This step can be performed in accordance with the above-described step I-1-2, and the same compound as that used in the above-described step I-1-2 can be used for the fluoroalcohol and λ3-iodide used in this step.

)

The small step Z-3 is as follows: next, the compound represented by formula D-3 is produced by reacting the compound represented by formula F-2 with 2, 2-trifluoro-N-phenyliminoacetyl chloride (TFPC).

The present process may be performed by using or applying a known method.

In one embodiment of the present invention, the small step Z-3 is the following step: the compound represented by the above formula D-3 is produced by reaction with 2, 2-trifluoro-N-phenyliminoacetyl chloride (TFPC) in the presence of N-methylimidazole, and is preferably produced, for example, by the method shown in example 36. As described in the same reaction in step I-1-3, the use of N-methylimidazole as the base used can reduce the equivalent weight of TFPC as compared with the case of potassium carbonate, for example, and can yield the target product in high yield. The solvent and the reaction temperature used, preferably in the presence of a dehydrating agent, and isolation/purification by column purification or the like are also carried out in the same manner as in the reaction in the above-mentioned steps I-1 to 3.

< Procedure II-2-2 >

The step II-2-2 is as follows: removing the protecting group phthalimide group of the amino group on the compound shown in the formula D-4 to obtain the compound shown in the formula D-5. This step can be preferably performed by the method shown in example 55-1, for example, by adding n-butanol and ethylenediamine to a solution containing a compound represented by formula D-4, but is not limited thereto.

< Purification of Compound (1) shown by formula D-5 >

In this step II-2-2, the compound represented by the formula D-5 can be obtained in a purified form by the following purification method. The purification method includes: after stopping the reaction between the compound represented by the formula D-3 and the compound represented by the formula D-4, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the produced compound represented by the formula D-5 and inclusions, the compound represented by the formula D-5 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and water, whereby the inclusions are removed, and then the compound represented by the formula D-5 is eluted from the hydrophobic carrier using an organic solvent, whereby the compound represented by the formula D-5 is purified. As described in the above-mentioned method for purifying the compound represented by the formula A-5 in the step I-1-1, the method for purifying can produce a large amount of high-quality oligosaccharides efficiently in the liquid phase synthesis of oligosaccharide chains by using a small amount of the hydrophobic carrier.

The purification of the compound represented by the formula D-5 is not limited to the purification in this step. Accordingly, in one aspect of the present invention, there is also provided a method comprising: adding a hydrophobic carrier and water to a water-soluble organic solvent containing the compound represented by the formula D-5 and impurities, adsorbing the compound represented by the formula D-5 to the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and water to remove the impurities, and eluting the compound represented by the formula D-5 from the hydrophobic carrier using the organic solvent to thereby purify the compound represented by the formula D-5.

The term "inclusion" refers to a compound or reagent other than a protecting oligosaccharide (a compound represented by formula D-5 in this step), and mainly refers to a reagent used in the synthesis reaction of a protecting oligosaccharide, a residue thereof, a sugar other than a protecting oligosaccharide such as a monosaccharide or disaccharide compound used in the extension reaction of a protecting oligosaccharide, or a by-product produced by the deprotection reaction of a protecting oligosaccharide. The "hydrophobic carrier" (resin for filling by reversed phase partition chromatography, etc.), "water-soluble organic solvent", "organic solvent", and the purification used in this step were the same as those described in the method for purifying the compound represented by formula A-5 in step I-1-1.

< Purification of Compound (2) represented by formula D-5 >

The compound represented by the above formula D-5 can be purified by the following steps. The purification may be performed separately from or in addition to the purification (1) of the compound represented by the above formula D-5. As this step, first, a compound represented by the following formula D-5-FMA, which is crystalline as a fumarate salt, is produced by reacting a compound represented by the formula D-5 with fumaric acid, whereby the compound represented by the crystalline formula D-5-FMA can be separated from a noncrystalline substance.

In the case of the compound represented by the formula D-5-FMA, the compound dissolved in the solvent may be used directly in the next step II-2-3 or may be converted into the compound represented by the formula D-5. The conversion to the compound represented by the formula D-5 can be carried out by the following method: the fumaric acid in the compound of formula D-5-FMA is removed to the aqueous layer by adding an aqueous alkaline solution, a solvent or the like, and then the organic layer is concentrated, whereby the compound of formula D-5 can be obtained in high purity. In this operation, the impurities having similar structures such as stereoisomers, which are difficult to remove even in column purification, can be easily removed. This step can be preferably performed by the method shown in example 55-2.

< Procedure II-2-3 >

The step II-2-3 is as follows: protecting the amino group in the compound represented by the above formula D-5 with a protecting group selected from the group consisting of aryloxycarbonyl (COOAr), acetyl (Ac), 2-trichloroethoxycarbonyl (Troc), and phthalimido (Pht) to give the compound represented by the above formula D-6 (wherein R ₅ is aryloxycarbonyl (COOAr), acetyl (Ac), or 2, 2-trichloroethoxycarbonyl (Troc), and R ₆ is a hydrogen atom, or R ₅ and R ₆ form a phthalimido group together with the nitrogen atom to which they are bonded). The purpose of introducing the protecting group for an amino group as described above is to make the target compound (the compound represented by the formula D-13) more efficient by taking the linear path of the acetyl group, but in the case where an amino group (-NHAc group) protected by an acetyl group is present in the reaction substrate in the glycosylation reaction between the compound represented by the formula D-6 and the compound represented by the formula D-7 in the subsequent step II-3, a significant decrease in the reactivity of the target glycosylation reaction is observed due to the interaction with the Lewis acid, and an excessive amount of sugar donor is required for the completion of the reaction. Therefore, the above-mentioned disadvantages can be avoided by protecting the temporary protecting group on the nitrogen of glucosamine by a protecting group selected from the group consisting of aryloxycarbonyl (COOAr), 2-trichloroethoxycarbonyl (Troc) and phthalimido (Pht) during the glycosylation reaction, and deprotecting the resulting product after the glycosylation reaction to form a-NHAc group. In addition, as the protecting group, an aryloxycarbonyl group (COOAr) is most preferably used. The "aryl (Ar)" in the aryloxycarbonyl group is a group generated by removing one hydrogen atom on an aromatic ring in an aromatic hydrocarbon, and is not limited, and examples thereof include phenyl, 2-naphthyl, 1-naphthyl, 2-pyridyl, 3-pyridyl, nitrophenyl, chlorophenyl, fluorophenyl, bromophenyl, iodophenyl, methoxyphenyl, and C1-C4 alkylphenyl, preferably phenyl. It has been found that aryloxycarbonyl (COOAr) allows glycosylation to proceed well as compared with other protecting groups, and that deprotection can be carried out under normal hydrolysis conditions, under preferable conditions such as room temperature and within 1 hour in the subsequent deprotection reaction.

The above-mentioned steps can be preferably performed by the methods shown in examples 56 to 59, for example, by adding tetrahydrofuran to a solution of the compound represented by the formula D-5 in tetrahydrofuran and dissolving sodium hydrogencarbonate, potassium hydrogencarbonate, disodium hydrogenphosphate, or dipotassium hydrogenphosphate in water, but are not limited thereto.

Instead of the above-mentioned steps II-2-2 and II-2-3, the compound represented by the above-mentioned formula D-6 can be produced by selectively removing the acetyl group (Ac) on the compound represented by the above-mentioned formula D-4 (wherein R ₅ and R ₆ together with the nitrogen atom to which they are bonded form a phthalimide group). The selective removal of acetyl groups may be performed under methyl trifluoroacetate conditions, but is not limited thereto. This step brings about the same results as the case where: in the steps II-2-2 and II-2-3, a phthalimide group (Pht) is selected as a protecting group for an amino group in the compound represented by the above formula D-5.

< Procedure II-3 >

The step II-3 is a step of producing a compound represented by the following formula D-11 (wherein M ⁺ is sodium ion, lithium ion, potassium ion, or protonated triethylamine cation), and includes the steps of: by forming β -1, 4-glycosidic bond with the compound represented by the above formula D-6 and the compound represented by the following formula D-7, the compound represented by the following formula D-8 is produced (wherein R ₅ is aryloxycarbonyl (COOAr), acetyl (Ac), or 2, 2-trichloroethoxycarbonyl (Troc), R ₆ is hydrogen atom, or R ₅ and R ₆ form phthalimide group together with nitrogen atom to which they are bonded), and then the protecting group of amino group in the compound represented by the formula D-8 is removed to produce the compound represented by the following formula D-9 (wherein M ⁺ is sodium ion, lithium ion, potassium ion, or triethylamine cation which is protonated).

In one embodiment of the present invention, the step II-3 includes the following steps II-3-1 to II-3-4.

< Procedure II-3-1 >)

The process comprises the following steps: the compound represented by the formula D-8 is produced by reacting the compound represented by the formula D-6 with a compound represented by the following formula D-7 to form a beta-1, 4-glycosidic bond. The glycoside bonding step may be performed by using or applying a known method, and is preferably performed by the methods shown in examples 60 to 63.

< Preparation of Compound represented by formula D-7 >

In one embodiment of the present invention, the compound represented by the formula D-7 can be produced as in the following small steps V-1 to V-11. This step includes a small step V-7 described below for synthesizing a disaccharide block by forming an α -2, 6-glycosidic bond with two monosaccharides as an essential small step, and may be performed by using a general method in the production of monosaccharides or oligosaccharides or by applying a general method.

In one embodiment of the present invention, the process V includes the following minor process.

)

The small step V-1 is as follows: the compound represented by the following formula G-2 was produced by protecting the hydroxyl group on the compound represented by the following formula G-1 with a benzoyl group.

The compound represented by the formula G-1, which is a starting material for the present step, is a compound designated as CAS number 100759-10-2, and can be produced by a known method, for example, by the methods shown in examples 37 and 38. The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 39.

)

The small step V-2 is as follows: the compound represented by the following formula G-3 was produced by detaching the benzylidene protecting group from the compound represented by the formula G-2.

The present step may be performed by using or applying a known method, and preferably may be performed by the method shown in example 40.

In one embodiment of the present invention, the present process includes the steps of: the compound represented by the formula G-3 is subjected to solid phase extraction by contacting a solvent in which the produced compound represented by the formula G-3 is dissolved with silica gel. Since the unreacted compound represented by the formula G-2 and the desorbed benzaldehyde do not adsorb on silica gel, the compound represented by the formula G-3 can be purified efficiently by this step.

Examples of the solvent used for dissolving the compound represented by the formula G-3 include toluene, heptane, methylene chloride, chloroform, or a combination thereof, preferably toluene, methylene chloride, chloroform, or a combination thereof, and particularly preferably toluene, but are not limited thereto.

The silica gel in this step is exemplified by a silica gel in an amount of 2 to 5 times the amount of the raw material, preferably a silica gel in an amount of 2 to 4 times the amount of the raw material, and more preferably a silica gel in an amount of about 3 times the amount of the raw material.

In this step, the solvent used for eluting the compound represented by the formula G-3 adsorbed on the silica gel is not particularly limited as long as it is a solvent in which the silica gel is insoluble and the target can be eluted, and examples thereof include cyclopentyl methyl ether, ethyl acetate, and t-butyl methyl ether.

)

The small step V-3 is as follows: the compound represented by the following formula G-5 was produced by esterifying a carboxylic acid of the compound represented by the following formula G-4, followed by injecting water.

The compound represented by the formula G-4 as the starting material in this step can be produced by a known method, or a commercially available product can be used. Examples of the commercial products of the compounds represented by the formula G-4 include N-acetylneuraminic acid produced by Tokyo chemical industry. The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 41.

)

The small step V-4 is as follows: a compound represented by the following formula G-6 is produced by selectively protecting a hydroxyl group other than the hydroxyl group to which the carbon at the 1-position in the compound represented by the formula G-5 is bonded with an acetyl group.

The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 42.

)

The small step V-5 is as follows: the compound represented by the following formula G-7 was produced by reacting a compound represented by the formula G-6 with 2, 2-trifluoro-N-phenyliminoacetyl chloride (TFPC).

The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 43.

In one embodiment of the present invention, the present step is a step of producing a compound represented by formula G-7 by reacting a compound represented by formula G-6 with TFPC in the presence of N-methylimidazole. As the base in this step, the equivalent weight of TFPC can be reduced in the case of using N-methylimidazole as compared with the case of using K ₂CO₃, and the target product can be obtained in high yield in this case as well. TFPC is an expensive reagent, so the yield improvement of this procedure is very beneficial in commercial production.

The reaction temperature in the present step is not limited as long as the reaction is carried out, and it is preferably 20 to 40 ℃, more preferably 10 to 35 ℃, particularly preferably 0 to 30 ℃.

)

The small step V-6 is as follows: the following compound represented by the formula G-8 was produced by protecting the nitrogen atom in the acetamido group of the compound represented by the formula G-7 with a t-butoxycarbonyl group.

The present step may be performed by using or applying a known method, and preferably may be performed by the method shown in example 44.

In this step, the compound represented by the formula G-8 thus produced may be used as it is in the next step, or may be isolated and purified by recrystallization. A great advantage of the compound represented by the formula G-8 is that isolation/purification can be performed by crystallization, and by crystallization, the compound represented by the formula G-8 having a purity of 99% or more in terms of HPLC can be obtained, and since it does not contain impurities, the glycosylation reaction in the next step can be stably carried out. Isolation/purification by recrystallization can be performed, for example, by adding heptane to a solution of cyclopentyl methyl ether to crystallize.

)

The small step V-7 is as follows: the compound represented by the following formula G-9 was produced by forming an alpha-2, 6-glycosidic bond between the compound represented by the formula G-8 and the compound represented by the formula G-3.

It is difficult to selectively bond a galactose derivative to an N-acetylneuraminic acid derivative through an α -2, 6-glycosidic bond, and for example, a method of synthesizing disaccharides by reacting a compound represented by the formula G-3 with a compound represented by the formula G-7 has been reported (j.org.chem., 2016, 81, 10600-10616), but reproduction of the reaction is not easily achieved, and the desired yield and selectivity cannot be obtained. In addition, this reaction has a problem that the selectivity decreases as the scale increases, the allowable reaction temperature range is also narrowed, and the influence of the reaction heat increases. The compound represented by the formula G-7, which is one of the starting compounds for the reaction, is very expensive, and thus low reproducibility, yield, selectivity in the reaction are a big problem in commercial production requiring scale-up. On the other hand, when the compound represented by the formula G-8 to which the t-butoxycarbonyl group is added is used as the starting compound in place of the compound represented by the formula G-7, high selectivity to the α -2, 6-glycosidic bond (α: β=93:7) can be realized with good reproducibility, and the yield is also improved. In addition, the allowable temperature range is also enlarged, and high reproducibility, yield and selectivity can be realized even when the scale is enlarged. This has an extremely beneficial effect on commercial production.

The present step may preferably be carried out in the presence of a lewis acid. The lewis acid in this step is not limited as long as the reaction proceeds, and examples thereof include: the trimethylsilyl triflate, triisopropylsilyl triflate, tert-butyldimethylsilyl triflate, and preferably trimethylsilyl triflate.

The solvent used in the present step is not limited as long as the reaction is performed, and examples thereof include diisopropyl ether, t-butyl methyl ether, diethyl ether, dibutyl ether, dipropyl ether, 1, 4-dioxane, methylene chloride, 1, 2-dichloroethane, toluene, chlorobenzene, trifluoromethylbenzene, propionitrile, and acetonitrile, and preferred examples thereof include cyclopentyl methyl ether.

The reaction temperature in the present step is not limited as long as the reaction is carried out, and examples thereof include-78℃to 0℃and preferably-78℃to-20℃and more preferably-78℃to-30℃and particularly preferably-78℃to-40 ℃.

In this step, it is preferable to add 1 to 3 equivalents of the compound represented by the formula G-3 to 1 equivalent of the compound represented by the formula G-8, and it is more preferable to add 1.4 to 2 equivalents of the compound represented by the formula G-3 to 1 equivalent of the compound represented by the formula G-8.

The present step is not limited as long as the reaction is performed, and for example, the step may be performed by adding dropwise a mixed solution of the compound represented by the formula G-8 and the compound represented by the formula G-3 (preferably a cyclopentyl methyl ether mixed solution) to a solution containing a lewis acid (preferably a cyclopentyl methyl ether solution) or adding dropwise a solution of the compound represented by the formula G-8 (preferably a cyclopentyl methyl ether solution) to a solution containing a lewis acid and the compound represented by the formula G-3 (preferably a cyclopentyl methyl ether solution) for a long period of time, and preferably may be performed by adding dropwise a solution of the compound represented by the formula G-8 (preferably a cyclopentyl methyl ether solution) to a solution containing a lewis acid and the compound represented by the formula G-3 (preferably a cyclopentyl methyl ether solution) for a long period of time. The time for the dropping is not limited as long as the reaction is performed, and is, for example, 30 minutes to 5 hours, preferably 1 hour to 4 hours, more preferably 2 hours to 3.5 hours, and particularly preferably about 3 hours.

In one embodiment of the present invention, the present process includes the steps of: the compound represented by formula G-9 is subjected to solid phase extraction by contacting a solvent in which the compound represented by formula G-9 is dissolved with silica gel. Since N-phenyltrifluoroacetamide, which is a byproduct in the glycosylation reaction, and other trace impurities in the toluene solvent which are not adsorbed on silica gel do not adsorb on silica gel, the compound represented by the formula G-9 can be purified efficiently by this step.

Examples of the solvent used for dissolving the compound represented by the formula G-9 include toluene, heptane, methylene chloride, chloroform, or a combination thereof, preferably toluene, methylene chloride, chloroform, or a combination thereof, and particularly preferably toluene, but are not limited thereto.

The silica gel in this step is exemplified by a silica gel in an amount of 2 to 5 times the amount of the raw material, preferably a silica gel in an amount of 2 to 4 times the amount of the raw material, and more preferably a silica gel in an amount of about 3.5 times the amount of the raw material.

In this step, the solvent used for eluting the compound represented by the formula G-9 adsorbed on the silica gel is not particularly limited as long as it is a solvent in which the silica gel is insoluble and the target can be eluted, and examples thereof include ethyl acetate, cyclopentyl methyl ether, and t-butyl methyl ether, and ethyl acetate is preferable.

This step can be performed by the method shown in example 45, for example.

)

The small step V-8 is as follows: a compound represented by the following formula G-10 is produced by detaching a t-butoxycarbonyl group from a compound represented by the formula G-9.

The present step may be performed by using or applying a known method, and preferably may be performed by the method shown in example 46.

)

The small step V-9 is as follows: the compound represented by the following formula G-11 was produced by further protecting the hydroxyl group of the compound represented by the formula G-10 and the nitrogen atom in the acetamido group with acetyl groups.

The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 47.

In one embodiment of the present invention, the present process includes the steps of: the compound represented by the formula G-11 is subjected to solid phase extraction by contacting a solvent in which the compound represented by the formula G-11 is dissolved with silica gel. Since the diacetyl or other by-product of the compound represented by the formula G-3, which is produced by acetylating the compound represented by the formula G-3 and used in excess in the upstream glycosylation reaction, is not adsorbed on silica gel, the compound represented by the formula G-11 can be purified efficiently by this step.

Examples of the solvent used for dissolving the compound represented by the formula G-11 include toluene, heptane, methylene chloride, chloroform, or a combination thereof, preferably toluene, methylene chloride, chloroform, or a combination thereof, and particularly preferably toluene, but are not limited thereto.

In this step, the solvent used for eluting the compound represented by the formula G-11 adsorbed on the silica gel is not particularly limited as long as it is a solvent in which the silica gel is insoluble and the target can be eluted, and examples thereof include ethyl acetate, cyclopentyl methyl ether, and t-butyl methyl ether, and ethyl acetate is preferable.

)

The small step V-10 is as follows: the compound represented by the following formula G-12 was produced by releasing an allyl group bonded to the 1-carbon of D-galactopyranoside in the compound represented by the formula G-11.

The present step may be performed by using or applying a known method, and preferably may be performed by the method shown in example 48.

In this step, the compound represented by the formula G-12 thus produced may be used as it is in the next step, or may be isolated and purified by recrystallization. A great advantage of the compound represented by the formula G-12 is that isolation/purification can be performed by crystallization, and by crystallization, the compound represented by the formula G-12 having a purity of 99% or more in terms of HPLC can be obtained, and since it does not contain impurities, the reaction in the next step can be stably carried out. Isolation/purification by recrystallization can be performed by, for example, a method of crystallizing by adding 2-propanol to a solution of ethyl acetate in which a compound represented by the formula G-12 is dissolved, and preferably, can be performed by, for example, a method shown in example 48.

)

The small step V-11 is as follows: the compound represented by the following formula D-7 was produced by reacting a compound represented by the formula G-12 with 2, 2-trifluoro-N-phenyliminoacetyl chloride (TFPC).

The present step may be performed by using or applying a known method, and is preferably performed by the method shown in example 49.

< Purification of Compound represented by formula D-8 >

In this step II-3-1, the compound represented by the formula D-8 can be obtained in a purified form by the following purification method. The purification method includes: after stopping the reaction between the compound represented by the formula D-6 and the compound represented by the formula D-7, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the produced compound represented by the formula D-8 and inclusions, the compound represented by the formula D-8 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and water, whereby the inclusions are removed, and then the compound represented by the formula D-8 is eluted from the hydrophobic carrier using an organic solvent, whereby the compound represented by the formula D-8 is purified. As described in the above-mentioned method for purifying the compound represented by the formula A-5 in the step I-1-1, the method for purifying can produce a large amount of high-quality oligosaccharides efficiently in the liquid phase synthesis of oligosaccharide chains by using a small amount of the hydrophobic carrier.

The purification of the compound represented by the formula D-8 is not limited to the purification in this step. Accordingly, in one aspect of the present invention, there is also provided a method comprising: the method comprises the steps of adding a hydrophobic carrier and water to a water-soluble organic solvent containing a compound represented by the formula D-8 and inclusions, adsorbing the compound represented by the formula D-8 on the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and water to remove the inclusions, and eluting the compound represented by the formula D-8 from the hydrophobic carrier using the organic solvent to thereby purify the compound represented by the formula D-8.

The term "inclusion" refers to a compound or reagent other than a protecting oligosaccharide (a compound represented by formula D-8 in this step), and mainly refers to a reagent used in the synthesis reaction of a protecting oligosaccharide, a residue thereof, a sugar other than a protecting oligosaccharide such as a monosaccharide or disaccharide compound used in the extension reaction of a protecting oligosaccharide, or a by-product produced by the deprotection reaction of a protecting oligosaccharide. The "hydrophobic carrier" (resin for filling by reversed phase partition chromatography, etc.), "water-soluble organic solvent", "organic solvent", and the purification used in this step were the same as those described in the method for purifying the compound represented by formula A-5 in step I-1-1.

< Procedure II-3-2 >

The process comprises the following steps: removing the protecting group of the amino group on the compound shown in the formula D-8 and the acyl protecting group of the alcohol to obtain the compound shown in the formula D-9. The removal (deprotection) of the protecting group of the amino group may be performed by using or applying a known method, and preferably may be performed by, for example, the method shown in example 64, and may be performed by sequentially adding 1, 2-dimethoxyethane and an aqueous solution of potassium hydroxide, sodium hydroxide, or lithium hydroxide, for example, but is not limited thereto.

The following steps II-3-3 to II-3-4 are exemplary embodiments for producing the compound represented by the formula D-11 from the compound represented by the formula D-9, but are not limited to these production steps.

< Procedure II-3-3 >

The process comprises the following steps: protecting the amino group on the compound represented by formula D-9 with an acetyl group to give the compound represented by the following formula D-10.

The protection of the amino group by acetyl group can be carried out by using or applying a known method, and preferably, it can be carried out by the method shown in example 65.

< Purification of Compound represented by formula D-10 >

In this step II-3-3, the compound represented by the formula D-10 can be obtained in a purified form by the following purification method. The purification method includes: adding a hydrophobic carrier and water to a water-soluble organic solvent containing the produced compound represented by the formula D-10 and impurities, adsorbing the compound represented by the formula D-10 to the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and water, thereby removing the impurities, and eluting the compound represented by the formula D-10 from the hydrophobic carrier using the organic solvent, thereby purifying the compound represented by the formula D-10. As described in the above-mentioned method for purifying the compound represented by the formula A-5 in the step I-1-1, the method for purifying can produce a large amount of high-quality oligosaccharides efficiently in the liquid phase synthesis of oligosaccharide chains by using a small amount of the hydrophobic carrier.

The purification of the compound represented by the formula D-10 is not limited to the purification in this step. Accordingly, in one aspect of the present invention, there is also provided a method comprising: the method comprises the steps of adding a hydrophobic carrier and water to a water-soluble organic solvent containing a compound represented by the formula D-10 and inclusions, adsorbing the compound represented by the formula D-10 on the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and water to remove the inclusions, and eluting the compound represented by the formula D-10 from the hydrophobic carrier using the organic solvent to thereby purify the compound represented by the formula D-10.

The term "inclusion" refers to a compound or reagent other than a protecting oligosaccharide (a compound represented by formula D-10 in this step), and mainly refers to a reagent used in the synthesis reaction of a protecting oligosaccharide, a residue thereof, a sugar other than a protecting oligosaccharide such as a monosaccharide or disaccharide compound used in the extension reaction of a protecting oligosaccharide, or a by-product produced by the deprotection reaction of a protecting oligosaccharide. The "hydrophobic carrier" (resin for filling by reversed phase partition chromatography, etc.), "water-soluble organic solvent", "organic solvent", and the purification used in this step were the same as those described in the method for purifying the compound represented by formula A-5 in step I-1-1.

< Procedure II-3-4 >

The process comprises the following steps: benzyl is removed from benzyloxy on the compound represented by formula D-10 to produce the compound represented by formula D-11. The above-mentioned benzyl group removal can be carried out by using or applying a known method, and preferably can be carried out by, for example, the method shown in example 66, for example, by adding N-methylpyrrolidone and Pd/C to the compound represented by the formula D-10, and then carrying out decompression, nitrogen substitution and hydrogen pressurization and decompression, but is not limited thereto.

< Procedure II-4 >

The step II-4 is as follows: comprising a step of reacting a compound represented by the formula D-11 with a compound represented by the following formula D-12 (11-azido-3, 6, 9-trioxaundecan-1-amine) as an azido PEG linker to produce an oligosaccharide represented by the formula D-13.

The bonding of the compound represented by the formula D-11 and the compound represented by the formula D-12 may be performed by using or applying a known method, and preferably may be performed, for example, by the method shown in example 72, for example, by sequentially adding the compound represented by the formula D-12, N-ethyldiisopropylamine, and hexafluorophosphoric acid (benzotriazol-1-yl-oxy) tripyrrolidinylphosphine, tripyrrolidinylphosphonium bromide hexafluorophosphate, or 4- (4, 6-dimethoxy-1, 3, 5-triazin-2-yl) -4-methylmorpholine hydrochloride to a solution containing the compound represented by the formula D-11, followed by stirring, but is not limited thereto.

< Purification of Compound represented by formula D-12 >

In one embodiment of the present invention, the compound represented by the formula D-12 is obtained by a purification method comprising the steps of: a step of adding a compound represented by the following formula E-1 (wherein R ₇ is a hydrogen atom, a methyl group, or a methoxy group) to a solution containing a crude compound represented by the formula D-12 to produce a crystalline compound represented by the following formula E-2 (wherein R ₇ is a hydrogen atom, a methyl group, or a methoxy group); isolating the crystalline compound, and then extracting the compound represented by formula D-12 from the isolated crystalline compound.

By this purification method, a compound represented by the formula D-12 having a high purity can be obtained, and the compound represented by the formula D-12 has a purity of preferably 95% or more, more preferably 96% or more, 97% or more, still more preferably 98% or more, or 99% or more, as measured by HPLC (also referred to as "HPLC purity" in the present specification). In this way, the purpose of purifying the compound represented by D-12 is to mix various other impurities represented by dimers into a commercially available reagent for the compound, and in the prior art, it is necessary to carry out strict distillation purification or complicated column purification, and in the case of containing an azide structure, there is a problem that the distillation operation requiring heating due to the explosion hazard cannot be applied. As a result of studies on a purification method for obtaining a compound represented by the formula D-12 having a high purity, the present inventors have found that when three tartaric acid derivatives represented by the formula E-1 (wherein R ₇ is a hydrogen atom, a methyl group, or a methoxy group) are used, the compound represented by the formula D-12 can form a 1-to-1 salt with these tartaric acid derivatives and isolate the salt as crystals. The compound represented by the above formula E-2 thus obtained is a novel crystalline compound, and after isolation, a compound represented by the formula D-12 having a higher HPLC purity (preferably 95% or more HPLC purity) than that before purification can be obtained by separating the compound with ethyl acetate/aqueous hydrochloric acid or the like, followed by ionization and extraction.

The refining method described above is described below as an exemplary method. First, a compound represented by the formula E-1 is added to a solution of a compound represented by the formula D-12 in a solvent such as acetonitrile and water, and after stirring and confirming dissolution, a solvent such as acetonitrile is added. The resulting slurry was concentrated under reduced pressure, and crystals precipitated by stirring the slurry were filtered. The filtered crystals were washed with acetonitrile and dried under reduced pressure to obtain crystals of the compound represented by formula E-2 (crystal-forming step). Then, concentrated hydrochloric acid is added to the solution of ethyl acetate and water of the crystalline compound obtained, and after stirring, the resultant solution is separated, the aqueous layer obtained is washed with ethyl acetate or the like, and after being made alkaline with an aqueous solution of sodium hydroxide or the like, sodium chloride or the like is added to dissolve the aqueous layer. Solvent such as dichloromethane was added, and after stirring, the resultant organic layer was separated and concentrated under reduced pressure. After adding a solvent such as acetonitrile, concentrating under reduced pressure, filtering the obtained solution, washing with a solvent such as acetonitrile, concentrating the obtained solution under reduced pressure (extraction step), the compound represented by formula D-12 having high purity by HPLC can be obtained. More preferably, the method is carried out by the methods shown in examples 67 to 71.

The purification of the compound represented by the formula D-12 is not limited to the purification in this step. Accordingly, in one aspect of the present invention, there is also provided a method for purifying a compound represented by formula D-12, the method comprising: a step of adding a compound represented by the formula E-1 (wherein R ₇ is a hydrogen atom, a methyl group, or a methoxy group) to a solution containing a crude compound represented by the formula D-12 to produce a crystalline compound represented by the formula E-2 (wherein R ₇ is a hydrogen atom, a methyl group, or a methoxy group); isolating the crystalline compound, and then extracting the compound represented by formula D-12 from the isolated crystalline compound.

< Novel Compounds >)

The intermediate of the oligosaccharide represented by the above formula A-13 is useful for the production of the oligosaccharide, but is not limited to the production of the oligosaccharide and can be applied to all applications. Thus, according to the present invention, there are provided an oligosaccharide represented by the above formula A-13 and an intermediate thereof.

In one embodiment of the present invention, there is provided an oligosaccharide represented by the following formula A-13.

In one embodiment of the present invention, there is provided a compound represented by the following formula A-5.

In one embodiment of the present invention, there is provided a compound represented by the following formula A-6.

In one embodiment of the present invention, there is provided a compound represented by the following formula A-7.

In one embodiment of the present invention, there is provided a compound represented by the following formula A-9.

In one embodiment of the present invention, there is provided a compound represented by the following formula A-10.

In one embodiment of the present invention, there is provided a compound represented by the following formula A-11.

In one embodiment of the present invention, there is provided a compound represented by the following formula A-12.

In one embodiment of the present invention, there is provided a compound represented by the following formula A-14.

In one embodiment of the present invention, there is provided a compound represented by the following formula A-15.

Furthermore, the intermediate of the compound represented by the above formula A-11 is useful for the production of the compound, but is not limited to the production of the compound and can be applied to all applications. Thus, according to the present invention, there is also provided an intermediate of the compound represented by the above formula A-11.

In one embodiment of the present invention, there is provided a compound represented by the following formula B-4.

In one embodiment of the present invention, there is provided a compound represented by the following formula B-5.

In one embodiment of the present invention, there is provided a compound represented by the following formula B-6.

In one embodiment of the present invention, there is provided a compound represented by the following formula B-7.

In one embodiment of the present invention, there is provided a compound represented by the following formula B-8.

Furthermore, the intermediate of the oligosaccharide represented by the above formula D-13 is useful for the production of the oligosaccharide, but is not limited to the production of the oligosaccharide and can be applied to all applications. As described below, according to the present invention, there are provided an oligosaccharide represented by the above formula D-13 and an intermediate thereof (also including a compound represented by the above formula A-13 and an intermediate thereof).

In one embodiment of the present invention, an oligosaccharide represented by the following formula D-13 is provided.

In one embodiment of the present invention, there is provided a compound represented by the following formula D-1.

Provided is a compound represented by the following formula D-2.

In one embodiment of the present invention, there is provided a compound represented by the following formula D-4.

In one embodiment of the present invention, there is provided a compound represented by the following formula D-5.

In one embodiment of the present invention, there is provided a compound represented by the following formula D-5-FMA.

In one embodiment of the present invention, there is provided a compound represented by the following formula D-6.

(Wherein R ₅ is an aryloxycarbonyl group (COOAr), an acetyl group (Ac), or a2, 2-trichloroethoxycarbonyl group (Troc), R ₆ is a hydrogen atom, or R ₅ and R ₆ form a phthalimide group together with the nitrogen atom to which they are bonded).

In one embodiment of the present invention, there is provided a compound represented by the following formula D-8.

In one embodiment of the present invention, there is provided a compound represented by the following formula D-9.

In one embodiment of the present invention, there is provided a compound represented by the following formula D-10.

In one embodiment of the present invention, there is provided a compound represented by the following formula D-11.

In one embodiment of the present invention, there is provided a crystalline compound represented by the following formula E-2.

(Wherein R ₇ is a hydrogen atom, a methyl group, or a methoxy group).

In one embodiment of the present invention, there is provided a compound represented by the following formula D-12 having a purity of 95% or more as measured by HPLC.

In one embodiment of the present invention, there is provided a compound represented by the above formula D-12 having a purity of 95% or more as measured by HPLC.

Glycoprotein and method for producing the same

In one embodiment of the present invention, there are provided a novel glycoprotein or the like using a double-antennary glycan (i.e., an oligosaccharide represented by formula D-13) having an α2, 6-sialic acid structure at the non-reducing end as a donor molecule for the synthesis of a glycoprotein or the like (particularly, a sugar chain-reconstituted antibody or a molecule comprising the FC region thereof, or an antibody drug conjugate), and a novel method for producing the same. As will be described in detail below, the oligosaccharide represented by the formula D-13 obtained by the production method of the present invention can be used for the production of glycoproteins (particularly, sugar chain-reconstituted antibodies or Fc region-containing molecules, or antibody drug conjugates) (WO 2019/065964, WO2020/050406, etc.), but is not limited thereto, and can be used for other applications.

In recent years, a method of reconstructing heterogeneous sugar chains of an antibody by an enzyme reaction and uniformly introducing sugar chains having functional groups has been reported (ACS chem. Biol.2012,7, 110-122,ACS Med.Chem.Lett.2016,7, 1005-1008). Attempts have been made to synthesize homogeneous Antibody Drug Conjugates (ADCs) using this sugar chain reconstitution technique, site-specifically introducing drugs (Bioconjugate chem.2015, 26, 2233-2242, angel.chem.int.ed.2016, 55, 2361-2367, us 2016361436).

Reconstruction of sugar chains first cleaves heterogeneous sugar chains added to a protein (antibody or the like) with hydrolase while leaving only terminal N-acetylglucosamine (GlcNAc), and a homogeneous protein fraction (hereinafter referred to as "acceptor molecule") to which GlcNAc is added is prepared. Subsequently, an arbitrary sugar chain (hereinafter referred to as "donor molecule") prepared separately is prepared, and the acceptor molecule and the donor molecule are linked using glycosyltransferase. Thus, a homogeneous glycoprotein having an arbitrary sugar chain structure can be synthesized.

In one embodiment of the present invention, the oligosaccharide represented by formula D-13 produced by the novel production method of the present invention can be used as a donor molecule in the synthesis of the above-mentioned homogeneous glycoprotein (particularly, the sugar chain-reconstructed antibody or the molecule comprising the Fc region thereof) by activating the terminal structure thereof.

Examples

In the examples below, room temperature is 15℃to 35 ℃. Silica gel chromatography was performed using Biotage Sfar HC D (20 μm, biotage) and reversed phase column chromatography was performed using Universal Column ODS Premium, 30, m L (manufactured by mountain Co., ltd.) and Inject column ODS L (manufactured by mountain Co., ltd.) and preparative HPLC was performed using AGILENT PREPARATIVE HPLC SYSTEM (manufactured by Agilent Technology). The columns were prepared using XBridge Prep OBD (5 μm, C18,250X 30mm, waters).

The following equipment was used for measurement of various spectral data. ¹ H-NMR and ¹³ C-NMR spectra were determined using ECZ500R and ECX400P from JEOL. Mass spectra were determined using Shimadzu LCMS-2010 and LCMS-2020 (manufactured by Shimadzu corporation), XEVO Q-Tof MS (Waters), Q-Exactive (Thermo Fisher).

< Synthesis of Compound represented by formula A-3 >

The compound represented by formula A-3 was synthesized according to the following synthesis pathway 1.

Synthesis path 1

Example 1

2-O-acetyl-3, 4, 6-tri-O-benzyl-D-mannopyranose (a compound represented by the formula A-2).

After adding 3,4, 6-tri-O-benzyl-1, 2-O- (1-methoxyethylene) - β -D-mannopyranose (compound represented by formula A-1) (40.0 g,78.9 mmol) to a 1L four-necked flask, ethyl acetate (400 mL) was added. Water (2 mL) and p-TsOH H ₂ O (45 mg,0.237 mmol) were added at room temperature under nitrogen atmosphere and stirred at the same temperature for 6 hours. After confirming the completion of the reaction by HPLC, triethylamine (7.99 g,78.9 mmol) was added thereto and stirred at the same temperature overnight. After completion of rearrangement of acetyl groups was confirmed by HPLC, 5% sodium bicarbonate water (400 mL) was added to the reaction solution to separate the solution. To the organic layer was added 20% saline (200 mL) and the mixture was separated. The organic layer was concentrated to 80mL under reduced pressure, toluene (400 mL) was added, and the mixture was concentrated to 80mL in vacuo. Toluene (400 mL) was added again and concentrated under reduced pressure to 80mL. Dehydrated toluene (120 mL) was added to obtain a toluene solution of 2-O-acetyl-3, 4, 6-tri-O-benzyl-D-mannopyranose (a compound represented by the formula A-2) as a colorless solution.

Example 2

2-O-acetyl-3, 4, 6-tri-O-benzyl-1-O- (2, 2-trichloroethane imido) -D-mannopyranose (Compound represented by formula A-3)

A toluene solution (78.9 mmol) of 2-O-acetyl-3, 4, 6-tri-O-benzyl-D-mannopyranose (a compound represented by formula A-2) was charged into a 1L flask, and trichloroacetonitrile (12 mL,118 mmol) and DBU (119. Mu.L, 0.789 mmol) were added. Stirring is carried out for 2 hours at 0℃under nitrogen. After completion of the reaction was confirmed by HPLC, acetic acid (45. Mu.L, 0.789 mmol) was added to the reaction solution at 0℃to obtain a toluene solution (78.9 mmol) of 2-O-acetyl-3, 4, 6-tri-O-benzyl-1-O- (2, 2-trichloroethane imido) -D-mannopyranose as a brown solution. The solution was used directly in the next step.

< Synthesis of Compound represented by formula A-11 >

The compound represented by formula A-11 was synthesized according to the following synthesis pathway 2.

[ Synthesis path 2]

Example 3

4-Methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- (2, 3,4, 6-tetra-O-acetyl-beta-D-galactopyranosyl) -beta-D-glucopyranoside (a compound represented by formula B-3)

To a solution of 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2-phthalimido-. Beta. -D-glucopyranoside (compound represented by the formula B-2) (2.508 kg,4.211 mol) in methylene chloride (17.5L), 2,3,4, 6-tetra-O-acetyl-1-O (2, 2-trichloro-imido) - α -D-galactopyranoside (compound represented by the formula B-1) (2.275 kg,4.618 mol) and a powder of molecular sieve 4A (10 μm or less, 375 g) were added, followed by stirring at 20℃to 30℃for 20 minutes, cooling to-20℃to-10℃and dropwise adding trimethylsilyltriflate (140 g,0.630 mol) over 3 minutes. After stirring at-5℃to-10℃for 3 hours, triethylamine (106 g,1.05 mol) was added, the temperature was raised to 0℃to 5℃and molecular sieve 4A was filtered off and washed with toluene (5L). After the obtained solution was concentrated under reduced pressure to 12.5L, toluene (12.5L) was added thereto, and the solution was subjected to 4-time liquid separation washing using a methanol (6.5L) -water (18.5L) mixed solution. The obtained organic layer was concentrated under reduced pressure to 9L, toluene (25L) was added, and then concentrated under reduced pressure to 7.5L, whereby a toluene solution (7.5L) of 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) -4-O- (2, 3,4, 6-tetra-O-acetyl-beta-D-galactopyranosyl) -beta-D-glucopyranoside (a compound represented by formula B-3) was obtained.

Example 4

4-Methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O-beta-D-galactopyranosyl-beta-D-glucopyranoside (a compound represented by formula B-4)

To a toluene solution (7.5L) of 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- (2, 3,4, 6-tetra-O-acetyl-. Beta. -D-galactopyranosyl) - -. Beta. -D-glucopyranoside (a compound represented by formula B-3), tetrahydrofuran (10L), methanol (5L) and methyl trifluoroacetate (178 g,4.20 mol) were added, respectively, followed by addition of a potassium t-butoxide-tetrahydrofuran solution (1M, 2.1L,2.1 mol) and stirring at 40℃to 45℃for 2 hours. After cooling to 20℃to 25℃acetic acid (151 g) and ethyl acetate (25L) were added, respectively, and washing was performed 3 times with sodium hydrogencarbonate (750 g) -sodium chloride (750 g) -water (20L) solution and 1 time with sodium chloride (2.5 kg) -water (10L) solution. The obtained organic layer was concentrated under reduced pressure to 7.5L, N-dimethylacetamide (25L) and cyclopentylmethyl ether (37.5L) were added, respectively, and after concentrating under reduced pressure to 27.5L, cyclopentylmethyl ether (12.5L) was added, and concentrating under reduced pressure to 27.5L, whereby N, N-dimethylacetamide solution (27.5L) of 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O-. Beta. -D-galactopyranoside (a compound represented by formula B-4) was obtained.

Example 5

4-Methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- (2, 3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl) -beta-D-glucopyranoside (a compound represented by formula B-5)

To a solution of t-butanol (1.56 kg,21.0 mol) in hexane (3.28 kg), a solution of n-butyllithium-hexane (15.2%, 8.88kg,21.0 mol) was added dropwise at-15℃to 0℃over 4 hours. Methyl trifluoroacetate (26.9 g,0.170 mol) was added to give a lithium tert-butoxide-hexane solution. To a solution (27.5L) of 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O-beta-D-galactopyranosyl-beta-D-glucopyranoside (a compound represented by formula B-4) in N, N-dimethylacetamide was added benzyl bromide (5.03 kg,29.4 mol) and molecular sieve 4A powder (less than 10 μm, 750 g), and a solution (12.1 kg) of lithium tert-butoxide in hexane was added dropwise over a period of 3 hours at 35℃to 45 ℃. After cooling to 20℃to 25℃acetic acid (378 g,6.29 mol) was added, and after filtration of molecular sieve 4A, it was washed with N, N-dimethylacetamide (7.5L). After washing with water (20L) by adding heptane (12.5L), tert-butyl methyl ether (25L) was added to the obtained N, N-dimethylacetamide layer, and washing with water (20L) was carried out 3 times. The obtained organic layer was concentrated under reduced pressure to 7.5L, whereby a tert-butyl methyl ether solution (7.5L) of 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- (2, 3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl) -beta-D-glucopyranoside (a compound represented by formula B-5) was obtained.

Example 6

4-Methoxyphenyl 3, 6-di-O-benzyl-2- (2-carboxybenzamide) -2-deoxy-4-O- (2, 3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl) -beta-D-glucopyranoside cinchonidine salt (compound represented by formula B-6)

Tetrahydrofuran (12.5L), methanol (5L) and water (1L) were added to a tert-butyl methyl ether solution (7.5L) of 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- (2, 3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl) -beta-D-glucopyranoside (a compound represented by formula B-5), and after cooling to 0℃to 10℃the mixture was cooled, a 4N aqueous sodium hydroxide solution (2.6L, 10.4 mol) was added dropwise over a period of 5 minutes. After stirring at 0℃to 10℃for 6 hours, triethylamine (1.70 kg,16.8 mol) was added thereto, and the mixture was stirred at 20℃to 30℃for 16 hours. After cooling to 0℃to 10℃it took 20 minutes to drop 6M hydrochloric acid (3.0L, 18.0 mol) and raise the temperature to 20℃to 25 ℃. Ethyl acetate (20L) was added thereto, followed by washing with water (17.5L) and then washing with sodium chloride (2.5 kg) -water (10L) solution. The obtained organic layer was concentrated under reduced pressure to 7.5L, and then ethyl acetate (17.5L) was added thereto and concentrated under reduced pressure to 7.5L. Ethyl acetate (30L) and cinchonidine (1.36 kg,4.62 mol) were added to the obtained solution, followed by stirring at 20℃to 25℃for 18 hours, cooling to 0℃to 5℃over 1 hour, and then dropwise adding heptane (20L) over 1 hour. After stirring for 1.5 hours at the temperature, the resulting crystals were filtered, and the resulting crystals were washed with a mixed solvent of ethyl acetate (7.5L) -heptane (5.6L) cooled to 0℃to 5 ℃. The obtained crystals were dried under reduced pressure at 40℃to obtain 4-methoxyphenyl 3, 6-di-O-benzyl-2- (2-carboxybenzamide) -2-deoxy-4-O- (2, 3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl) -beta-D-glucopyranoside cinchonidine salt (compound represented by formula B-6) (5.10 kg, total yield of 4 steps 85.0%).

¹H－NMR(500MHz,CDCl₃)δ8.78(d,4.6Hz,1H),8.75(d,J＝8.0Hz,1H),8.07(d,J＝8.5Hz,1H),8.06(d,J＝8.0Hz,2H),7.65－7.68(m,3H),7.61(dd,3.9,7.7Hz,1H),7.55(d,J＝4.6Hz,1H),7.43(dd,3.7,7.4Hz,2H),7.19－7.33(m,28H),6.98－7.04(m,5H),6.68－6.70(m,2H),6.32(brd,1H),5.39－5.46(m,1H),5.35(d,J＝6.9Hz,1H),4.97(d,J＝11Hz,1H),4.83－4.91(m,3H),4.66－4.73(m,5H),4.55(d,J＝12Hz,1H),4.46(s,1H),4.43(d,J＝4.0Hz,1H),4.35(dd,J＝5.7,12Hz,2H),4.26(d,J＝12Hz,1H),4.22(dd,J＝4.2,8.4Hz,1H),4.03－4.09(m,3H),3.92(d,J＝2.6Hz,1H),3.74(d,J＝3.7Hz,2H),3.67－3.70(m,4H),3.57－3.60(m,1H),3.54(t,J＝7.9Hz,1H),3.36－3.44(m,3H),3.31(t,J＝9.2Hz,1H),3.12(dd,10,14Hz,1H),3.02(d,J＝13Hz,1H),2.93(m,1H),2.36(s,1H),1.86－1.89(m,2H),1.81(dd,J＝9.0,13Hz,1H),1.53(dd,J＝4.7,9.5Hz,1H),1.09(dd,J＝5.7,11Hz,1H).

¹³C－NMR(125MHz,CDCl₃)δ176.2,169.5,155.3,151.6,150.1,148.0,146.4,138.88,138.7,138.53,138.48,138.39,138.35,138.26,137.8,133.4,130.15,130.07,129.11,128.73,128.70,128.51,128.4,128.34,128.22,128.2,128.16,127.95,127.92,127.83,127.75,127.57,127.55,127.43,127.41,127.19,127.12,124.9,122.8,119.5,118.7,116.2,114.3,103.2,100.3,82.4,79.8,78.7,76.2,75.2,75.1,74.8,73.7,73.4,73.03,72.99,72.7,68.7,68.1,59.8,55.5,55.3,54.1,43.4,37.6,27.0,25.0,19.1.

HRMS (ESI ⁺)[M+HNEt₃]⁺C₇₅H₈₅N₂O₁₄ theory: 1237.5995; experimental 1237.5977).

[α]_D ²⁰＝－21.889(c 1.003,CDCl₃)。

Example 7

To a suspension of 4-methoxyphenyl 3, 6-di-O-benzyl-2- (2-carboxybenzamide) -2-deoxy-4-O- (2, 3,4, 6-tetra-O-benzyl-. Beta. -D-galactopyranosyl) - β -D-glucopyranoside cinchonidine salt (compound of formula B-6) (4.50 kg,3.15 mol) in ethyl acetate (33.8L) was added 0.5M hydrochloric acid (33.8L, 16.9 mol) at 15℃to dissolve it under stirring. After the aqueous layer was removed, a liquid-separating washing with a sodium chloride (4.5 kg) -water (18L) solution was performed, and the obtained organic layer was concentrated to 6.8L under reduced pressure. After ethyl acetate (33.8L) was added, the mixture was concentrated under reduced pressure to 6.8L, and the compound represented by formula B-7 was obtained as an ethyl acetate solution.

Next, tetrahydrofuran (18L) was added to the obtained solution. After cooling to 0℃to 5℃1,1' -carbonyldiimidazole (765 g,4.12 mol) was added thereto and stirred for 17 hours. After ethyl acetate (22.5L), water (22.5L) and 6M hydrochloric acid (1.57L, 9.42 mol) were added, respectively, the aqueous layer was removed. The obtained organic layer was washed with water (22.5L) and sodium chloride (4.5 kg) -water (18L) solution in this order. After the obtained organic layer was concentrated under reduced pressure to 4.5L, toluene (22.5L) was added thereto and concentrated under reduced pressure to 4.5L, whereby a toluene solution (4.5L) of 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) -4-O- (2, 3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl) - β -D-glucopyranoside (a compound represented by formula B-5) was obtained.

Example 8

3, 6-Di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- (2, 3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl) -beta-D-glucopyranose (Compound represented by formula B-8)

To a toluene solution (4.5L) of 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- (2, 3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl) -beta-D-glucopyranoside (a compound represented by formula B-5) were added methylene chloride (6.75L), 1, 3-hexafluoro-2-propanol (6.75L), and water (675 mL), respectively, at 15℃to 25 ℃. A suspension of bis (trifluoroacetoxy) iodobenzene (2.16 kg,5.02 mol) in methylene chloride (6.75L) was added in 10 parts and stirred at 15℃to 25℃for 20 hours. After cooling to 0 to 5℃and adding toluene (31.5L), the mixture was washed 2 times with a solution of sodium hydrogencarbonate (900 g) -sodium sulfite (900 g) -water (22.5L) and washed at 0 to 20 ℃. The organic layer was washed with a sodium chloride (4.5 kg) -water (18L) solution, concentrated under reduced pressure to 9L, and toluene (22.5L) was added and concentrated under reduced pressure to 9L. Toluene (36L) was added to the resulting solution, and the mixture was divided into 2 parts. Silica gel (4.5 kg) was added to each of the divided solutions, followed by stirring at 20℃to 25℃for 3 hours, followed by filtration of the silica gel and washing with toluene (45L). The silica gel obtained was desorbed from the silica gel by washing with ethyl acetate (7.5L) -toluene (22.5L) -mixed solution. The obtained organic layers were mixed and concentrated under reduced pressure to 9L, whereby a toluene solution (9L) of 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- (2, 3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl) -beta-D-glucopyranose (a compound represented by the formula B-8) was obtained.

¹H－NMR(500MHz,CDCl₃)δ7.60－7.87(m,4H),7.18－7.36(m,25H),6.96(d,J＝6.9Hz,2H),6.89(m,1H),6.83(dd,J＝7.4,10Hz,2H),5.33(dd,J＝1.9,3.7Hz,0.2H),5.30(dd,J＝4.3,8.6Hz,0.8H),4.92(dd,J＝9.9,12Hz,2H),4.78－4.86(m,2H),4.71(d,J＝2.3Hz,2H),4.44－4.61(m,3H),4.36－4.14(m,3H),4.34(d,J＝4.6Hz,1H),4.26(d,J＝12Hz,1H),4.03－4.12(m,2H),3.89(d,J＝2.8Hz,1H),3.85(dd,J＝4.0,11Hz,1H),3.77(dd,J＝7.7,9.7Hz,1H),3.67(dd,J＝1.6,11Hz,1H),3.56－3.60(m,1H),3.35－3.49(m,4H),2.88(d,J＝8.6Hz,1H).

¹³C－NMR(125MHz,CDCl₃)δ168.2,139.06,139.04,138.98,138.91,138.7,138.6,138.5,138.10,138.06,133.8,131.7,128.40,128.38,128.32,128.27,128.24,128.10,127.99,127.96,127.90,127.84,127.78,127.76,127.72,127.68,127.65,127.63,127.55,127.51,127.49,127.44,127.42,127.3,126.8,123.6,123.3,102.9,102.8,93.1,92.8,82.34,82.32,80.02,79.99,77.8,77.7,76.6,75.43,75.39,75.31,74.53,74.49,74.33,73.98,73.72,73.63,73.41,73.24,73.20,73.07,72.63,72.59,70.6,68.3,68.0,67.8,57.7,55.8.

HRMS (ESI ⁺)[M+Na]⁺C₆₂H₆₁NNaO₁₂ theory: 1034.4092; experimental 1034.4071).

[α]_D ²⁰＝+33.243(c 1.002,CDCl₃)。

Example 9

3, 6-Di-O-benzyl-2-deoxy-2- (1, 3-Dioxo-1, 3-dihydro-2H-isoindol-2-yl) -4-O- (2, 3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl) -1-O- (2, 2-trifluoro-N-phenylethanimido) -beta-D-glucopyranose (a compound represented by formula A-11)

To a toluene solution (total amount: 1002 g) of 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- (2, 3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl) -beta-D-glucopyranose (a compound represented by formula B-8) (a solution obtained from 500g, 0.349mol of examples 7, 8 derived from the compound represented by formula B-6) were added dichloromethane (2.0L), molecular sieve 4A powder (10 μm or less, 250 g) and N-methylimidazole (34.4 g, 0.319 mol) at 20℃to 30 ℃.2, 2-trifluoro-N-phenyliminoacetyl chloride (79.8 g,0.384 mol) was then added thereto, and the mixture was stirred at 20℃to 30℃for 17 hours. After molecular sieve 4A was filtered, it was washed with toluene (500 mL) and the resulting solution was cooled to 0℃to 10 ℃. The column was packed with silica gel (1.5 kg) wet with methylene chloride cooled to 0℃to 5℃and washed with methylene chloride (15L) cooled to 0℃to 5℃and fractionated every 2.5L to 3L to obtain a fraction. Next, the mixture was washed with a mixture of methylene chloride (10L) containing 3% ethyl acetate, and the mixture was fractionated every 2.5L to 3L to obtain a fraction. The 1 st to 7 th fractions were collected and concentrated under reduced pressure to 1L to give a toluene-methylene chloride mixed solution of 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- (2, 3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl) -1-O- (2, 2-trifluoro-N-phenylethanimido) -beta-D-glucopyranose (compound represented by formula A-11). The solution was used directly in example 27.

¹H－NMR(500MHz,CDCl₃)δ7.62－7.82(m,4H),7.01－7.53(m,31H),6.98－7.01(m,1H),6.95(d,J＝7.2Hz,2H),6.80－6.88(m,3H),6.64(brd,2H),4.93(d,J＝12Hz,1H),4.89(d,J＝12Hz,1H),4.83(d,J＝11Hz,1H),4.77(d,J＝11Hz,1H),4.67－4.72(m,2H),4.56(d,J＝12Hz,1H),4.53(d,J＝12Hz,1H),4.36－4.48(m,5H),4.28(d,J＝12Hz,1H),4.14(t,J＝8.7Hz,1H),3.89(d,J＝2.6Hz,1H),3.85(d,J＝7.7Hz,1H),3.76(t,J＝8.4Hz,1H),3.38－3.50(m,4H).

¹³C－NMR(125MHz,CDCl₃)δ167.6,143.4,143.1,139.0,138.68,138.66,138.5,137.08,137.06,135.2,133.92,131.5,129.3,129.1,129.0,128.5,128.41,128.37,128.31,128.27,128.22,128.90,127.86,127.69,127.56,127.45,127.44,127.2,126.9,126.2,124.3,123.4,120.6,120.5,119.3,116.3(q,J＝148.5Hz),102.9,93.5,82.3,79.9,76.6,76.0,75.4,74.5,73.61,73.41,73.08,73.05,72.6,68.3,67.2,54.8.

HRMS (ESI ⁺)[M+Na]⁺C₇₀H₆₅F₃N₂NaO₁₂ theory: 1205.4387; experimental 1205.43835).

[α]_D ²⁰＝+66.645(c 1.099,CDCl₃)。

< Synthesis of Compound represented by formula A-13 >

The compound represented by the formula A-13 was synthesized according to the following synthesis pathway 3.

Synthesis path 3

Example 10

1, 2:5, 6-Bis-O- (1-methylethylidene) -3-O- (2-naphthylmethyl) -alpha-D-furanose (Compound represented by formula C-2)

After cooling a solution of sodium hydride (55.32 g,1.38mol, content: 50% -72%) in tetrahydrofuran (900 mL) to 0 ℃, a solution of 1, 2:5, 6-bis-O- (1-methylethylene) - α -D-furanose (a compound represented by the formula C-1) (300.00 g,1.15 mol) in tetrahydrofuran (1.05L) was added dropwise over 1 hour. Thereafter, the temperature was raised to 25℃and 1, 3-dimethyl-2-imidazolidinone (150 mL) and 2-bromomethylnaphthalene (280.31 g,1.27 mol) were added. After stirring at 25℃for 6 hours, the end of the reaction was confirmed by HPLC, ethylenediamine (anhydrous) (13.85 g,230.52 mmol) was added, and stirring was further performed for 1 hour. The solution was cooled to 0℃and 10% aqueous citric acid (1.2L) was added over 1 hour. The reaction solution was diluted with heptane (3L), and separated into an organic layer and an aqueous layer. After washing the organic layer with water (900 mL), the solution was concentrated under reduced pressure until the amount of the solution reached 900mL. Acetonitrile (3L) was further added and concentrated again to a liquid volume of 900mL to give the crude product 1, 2:5, 6-bis-O- (1-methylethylidene) -3-O- (2-naphthylmethyl) - α -D-furanglucose (compound represented by formula C-2) as an acetonitrile solution. The product was used directly in the next step.

Example 11

3-O- (2-naphthylmethyl) -D-glucopyranose (Compound represented by formula C-3)

To a solution (900 mL) of the compound represented by the formula C-2 as the crude product obtained in example 10 were added acetonitrile (1.5L), water (600 mL) and concentrated hydrochloric acid (17.51 g,172.89 mmol), and the mixture was stirred at 55℃for 18.5 hours. After completion of the reaction was confirmed by HPLC, the reaction solution was cooled to 0℃and the pH in the system was adjusted to 6.25 with 4N aqueous sodium hydroxide solution (43.22 mL). The reaction was diluted with heptane (900 mL) and separated into an acetonitrile layer and a heptane layer. Ethyl acetate (2.4L) and water (600 mL) were added to the acetonitrile layer to separate the liquid, thereby obtaining an organic layer A and an aqueous layer. A mixed solution of ethyl acetate (1.5L) and tetrahydrofuran (1.5L) was added to the aqueous layer again to separate the liquid, thereby obtaining an organic layer B and an aqueous layer. The organic layers A and B were mixed, washed with saturated brine (600 mL), and concentrated under reduced pressure until the liquid volume reached 1.5L (crystal deposition was confirmed in the concentration step). Ethyl acetate (4.5L) was further added, and the mixture was concentrated again until the liquid amount reached 3L. To the suspension were added ethyl acetate (1.5L) and cyclopentyl methyl ether (1.5L), and the mixture was stirred at 55℃for 1 hour. Heptane (3L) was added dropwise over 1.5 hours, stirred for 1 hour and then cooled to 0 ℃. Thereafter, the precipitated crystals were filtered, and the crystals were washed with a mixed solution of ethyl acetate (1.2L) and heptane (600 mL) cooled to 0 ℃. The obtained crystal was dried under reduced pressure at 40℃to obtain 3-O- (2-naphthylmethyl) -D-glucopyranose (compound represented by the formula C-3) (356.95 g, yield 96.7%).

¹H－NMR(500MHz,DMSO－d₆)δ7.85－7.90(m,4H),7.59(dd,J＝8.0,1.5Hz,1H),7.46－7.51(m,2H),6.69(d,J＝6.0Hz,1H),5.12(dd,J＝5.0,3.0Hz,2H),4.94－5.00(m,2H),4.53(t,J＝6.0Hz,1H),4.35(dd,J＝8.0,6.5Hz,1H),3.70(ddd,J＝11.5,5.0,2.0Hz,1H),3.45－3.50(m,1H),3.25－3.31(m,2H),3.11－3.16(m,2H).

¹³C－NMR(125MHz,DMSO－d₆)δ137.3,132.8,132.3,127.6,127.5,127.4,126.1,126.0,125.6,125.5,96.9,85.4,76.7,74.8,73.7,69.9,61.1.

HRMS (ESI ^－)[M－H]^－C₁₇H₂₀O₆ theory: 320.1260; experimental 319.1175).

Example 12

2,4, 6-Tri-O-acetyl-3-O- (2-naphthylmethyl) -D-glucopyranose (Compound represented by formula C-5)

To a solution of 3-O- (2-naphthylmethyl) -D-glucopyranose (compound represented by formula C-3) (150.00 g,468.25 mmol) in tetrahydrofuran (675 mL) were added triethylamine (236.92 g,2.34 mmol) and 4-dimethylaminopyridine (0.29 g,2.34 mmol), and after cooling to 0℃acetic anhydride (195.99 g,1.92 mol) was added dropwise over a period of 30 minutes. After that, the temperature was raised to 25℃and stirred for 3 hours, and the completion of the reaction was confirmed by HPLC. The reaction solution was cooled to 10℃and 1-methylpiperazine (60.97 g,608.73 mmol) was added. After stirring at 35 ℃ for 18 hours, the end of the reaction was confirmed by HPLC and cooled to 0 ℃. After adjusting the pH to 6.36 with 6N hydrochloric acid (480 mL), the mixture was diluted with heptane (375 mL) and the organic and aqueous layers were separated. After washing the organic layer with saturated aqueous sodium bicarbonate (450 mL) and water (450 mL), the solution was concentrated under reduced pressure until the amount of liquid reached 450mL. Ethyl acetate (2.25L) was added thereto, and the mixture was concentrated again to a liquid volume of 450mL, and the same procedure was repeated once more. To this solution was added methylene chloride (2.25L), and the solution was concentrated to a liquid volume of 450mL, and the same operation was repeated once more, whereby a crude product of 2,4, 6-tri-O-acetyl-3-O- (2-naphthylmethyl) -D-glucopyranose (a compound represented by the formula C-5) was obtained as a methylene chloride solution. The product was used directly in the next step.

The step of converting the compound represented by the formula C-3 to the compound represented by the formula C-5 was performed by a one-pot method.

Example 13

2,4, 6-Tri-O-acetyl-3-O- [ (naphthalen-2-yl) methyl ] -1-O- (2, 2-trichloroethane imido) -D-glycerol-hexopyranose (Compound represented by formula C-6)

To a solution (450 mL) of the crude product 2,4, 6-tri-O-acetyl-3-O- (2-naphthylmethyl) -D-glucopyranose (compound represented by formula C-5) obtained in example 12 were added dichloromethane (450 mL) and trichloroacetonitrile (338.03 g,2.34 mol), and after cooling to 0 ℃,1, 8-diazabicyclo [5.4.0] -7-undecene (5.70 g,37.46 mmol) was added dropwise. After stirring at 0℃for 14.5 hours, the end of the reaction was confirmed by HPLC and acetic acid (2.25 g,37.46 mmol) was added. Silica gel 60N (particle size: 40 to 50 μm,150g, manufactured by Kato chemical Co., ltd.) was added to the solution, and the mixture was stirred for 1.5 hours and then filtered. The silica gel was washed with methylene chloride (1.5L), and the filtrate was concentrated under reduced pressure to a liquid volume of 450mL. Further, methylene chloride (1.5L) was added thereto and concentrated to a liquid volume of 450mL, whereby 2,4, 6-tri-O-acetyl-3-O- [ (naphthalen-2-yl) methyl ] -1-O- (2, 2-trichloroethane imido) -D-glyceropyranose (a compound represented by the formula C-6) was obtained as a methylene chloride solution. The product was used directly in the next step.

Example 14

4-Methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- {2,4, 6-tri-O-acetyl-3-O- [ (naphthalen-2-yl) methyl ] -beta-D-glucopyranosyl } -beta-D-glucopyranoside (a compound represented by formula C-8)

To a solution (450 mL) of 2,4, 6-tri-O-acetyl-3-O- [ (naphthalen-2-yl) methyl ] -1-O- (2, 2-trichloroethaneimido) -D-glycero-hexopyranose (compound represented by formula C-6) obtained in example 13 was added 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (compound represented by formula C-7) (276.66 g,468.25 mmol), methylene chloride (4.2L) and a powder (10 μm or less, 83.00 g) of molecular sieve 4A, and the mixture was cooled to-5 ℃. To the suspension was added dropwise trimethylsilyl triflate (10.41 g,46.83 mmol) over 20 minutes, followed by stirring for 3 hours. After confirming the end of the reaction by HPLC, triethylamine (23.69 g,234.13 mmol) was added. After the suspension was filtered, it was washed with ethyl acetate (2.8L), and the filtrate was concentrated under reduced pressure to a liquid amount of 1.4L. Further, ethyl acetate (4.2L) was added thereto, and the mixture was concentrated to a liquid volume of 1.4L, and the same procedure was repeated once more. To the solution was added ethyl acetate (2.8L), which was washed with saturated aqueous sodium hydrogencarbonate solution (830 mL) and water (830 mL), and the organic layer was concentrated under reduced pressure to a liquid volume of 830mL. 2-propanol (4.2L) was further added, and after concentrating to a liquid volume of 1.4L, the suspension was adjusted to 65 ℃. Ethyl acetate (830 mL) was added and, after stirring at 65℃for 2 hours, 2-propanol (5.53L) was added dropwise over 2 hours. After cooling the suspension to 0 ℃, the crystals were filtered and washed with 2-propanol (1.4L) cooled to 0 ℃. The resulting crystals were dried under reduced pressure at 40℃to give a crude product of 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- {2,4, 6-tri-O-acetyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-glucopyranosyl } - β -D-glucopyranoside (compound represented by formula C-8) (355.34 g, yield 90.5%, based on the compound represented by formula C-3).

To the obtained crude compound of formula C-8 (350.00 g) was added methyl isobutyl ketone (2.1L), and after dissolution at 50℃ethyl cyclohexane (1.4L) was added dropwise over 1 hour. 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- {2,4, 6-tri-O-acetyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-glucopyranosyl } - β -D-glucopyranoside (a compound represented by formula C-8) (70.00 mg) is added and stirred for 1 hour, after confirming precipitation of crystals, ethylcyclohexane (4.9L) is added dropwise over 2 hours. The suspension was cooled to room temperature, stirred for 14.5 hours, and then the precipitated crystals were filtered, and the crystals were washed with a mixed solution of methyl isobutyl ketone (350 mL) and ethylcyclohexane (1.4L). The obtained crystals were dried under reduced pressure at 40℃to give 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- {2,4, 6-tri-O-acetyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-glucopyranosyl } - β -D-glucopyranoside (compound represented by formula C-8) (331.74 g, yield 94.8%).

¹H－NMR(500MHz,CDCl₃)δ7.81－7.84(m,4H),7.69(br,1H),7.65(br,3H),7.46－7.51(m,2H),7.28－7.36(m,6H),7.00(dd,J＝7.0,1.5Hz,2H),6.77－6.84(m,5H),6.66－6.69(m,2H),5.59,(d,J＝9.0Hz,1H),5.09－5.15(m,2H),4.82(d,J＝12.5Hz,1H),4.77(d,J＝12.0Hz,1H),4.71－4.77(m,2H),4.60(d,J＝8.0Hz,1H),4.50(d,J＝12.5Hz,1H),4.45(d,J＝13.0Hz,1H),4.36(dd,J＝11.0,8.5Hz,1H),4.28(dd,J＝11.0,8.5Hz,1H),4.20(dd,J＝12.5,5.0Hz,1H),4.10(dd,J＝10.0,8.5Hz,1H),3.99(dd,J＝12.0,2.0Hz,1H),3.80(br,2H),3.69(s,3H),3.58－3.62(m,2H),3.44(ddd,J＝10.0,4.5,2.5Hz,1H),1.98(s,3H),1.938(s,3H),1.937(s,3H).

¹³C－NMR(125MHz,CDCl₃)δ171.0,169.5,169.1,155.6,151.0,138.7,138.2,135.5,133.9,133.4,133.2,128.7,128.4,128.23,128.20,128.06,128.05,127.9,127.2,126.5,126.2,125.7,123.5,118.9,114.5,100.7,97.8,80.6,78.5,76.8,75.2,74.8,74.1,73.8,73.1,72.1,69.9,67.8,62.2,55.78,55.75,21.1,20.94,20.85.

HRMS (ESI ⁺)[M+H]⁺C₅₈H₅₈NO₁₆ theory: 1024.3750; experimental 1024.3706).

The spectra were confirmed to be consistent with the following: org.biomol.chem.,2018, 16, 4720-4727.

Example 15

4-Methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- {3-O- [ (naphthalen-2-yl) methyl ] - β -D-glucopyranosyl } - β -D-glucopyranoside (a compound represented by formula C-9)

To a solution of 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- {2,4, 6-tri-O-acetyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-glucopyranosyl } - β -D-glucopyranoside (compound of formula C-8) (30.00 g,29.29 mmol) in tetrahydrofuran (150 mL) were added methanol (90 mL) and methyl trifluoroacetate (3.75 g,29.29 mmol), and after stirring at 25℃for 10 minutes, potassium tert-butoxide (1 mol/L in tetrahydrofuran) (14.7 mL,14.65 mmol) was added. After that, the temperature was raised to 55℃and stirred for 2 hours, and the completion of the reaction was confirmed by HPLC. The reaction solution was cooled to 25℃and acetic acid (1.76 g,29.29 mmol) and ethyl acetate (300 mL) were added sequentially. After washing the solution twice with 1% aqueous sodium chloride (300 mL), the solution was concentrated under reduced pressure to a liquid volume of 90mL. Ethyl acetate (450 mL) was added, concentrated again to a liquid volume of 90mL, acetonitrile (450 mL) was further added, concentrated to a liquid volume of 90mL, and 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) -4-O- {3-O- [ (naphthalen-2-yl) methyl ] - β -D-glucopyranosyl } - β -D-glucopyranoside (a compound represented by formula C-9) was obtained as an acetonitrile solution. The product was used directly in the next step.

Example 16

4-Methoxyphenyl 3, 6-di-O-benzyl-4-O- {4, 6-O-benzylidene-3-O- [ (naphthalen-2-yl) methyl ] -beta-D-glucopyranosyl } -2-deoxy-2- (1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranoside (a compound represented by formula C-10)

To a solution (90 mL) of 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- {3-O- [ (naphthalen-2-yl) methyl ] - β -D-glucopyranosyl } - β -D-glucopyranoside (a compound represented by formula C-9) were added acetonitrile (210 mL), benzaldehyde dimethyl acetal (5.13 g,33.69 mmol) and p-toluenesulfonic acid monohydrate (0.17 g,0.88 mmol), and stirred at 25℃for 30 minutes. Toluene (600 mL) was added to the solution, and the solution was concentrated to 300mL, at which point the completion of the reaction was confirmed by HPLC. 1-methylimidazole (12.03 g,146.47 mmol) was further added and concentrated to a liquid volume of 90mL, to obtain 4-methoxyphenyl 3, 6-di-O-benzyl-4-O- {4, 6-O-benzylidene-3-O- [ (naphthalen-2-yl) methyl ] - β -D-glucopyranosyl } -2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (a compound represented by formula C-10) as a toluene solution containing 1-methylimidazole. The product was used directly in the next step.

Example 17

4-Methoxyphenyl 3, 6-di-O-benzyl-4-O- {4, 6-O-benzylidene-3-O- [ (naphthalen-2-yl) methyl ] -2-O- (trifluoromethanesulfonyl) - β -D-glucopyranosyl } -2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (a compound represented by formula C-11 (wherein X ₁ is Tf group))

To a solution (90 mL, 1-methylimidazole-containing ethyl acetate (210 mL) of the 4-methoxyphenyl 3, 6-di-O-benzyl-4-O- {4, 6-O-benzylidene-3-O- [ (naphthalen-2-yl) methyl ] - β -D-glucopyranosyl } -2-deoxy-2- (1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (compound represented by formula C-10) obtained in example 16 was added, and the mixture was cooled to 0 ℃. To this solution, trifluoromethanesulfonic anhydride (16.53 g,58.59 mmol) was added dropwise over 1 hour, followed by stirring for 30 minutes. After completion of the reaction was confirmed by HPLC, water (300 mL) was added, and the mixture was separated into an organic layer and an aqueous layer. The organic layer was washed twice with water (300 mL), once with a saturated aqueous solution of sodium chloride (150 mL), and then concentrated under reduced pressure to a liquid volume of 90mL. Ethyl acetate (300 mL) was added and the mixture was concentrated again to a liquid volume of 90mL, and 4-methoxyphenyl 3, 6-di-O-benzyl-4-O- {4, 6-O-benzylidene-3-O- [ (naphthalen-2-yl) methyl ] -2-O- (trifluoromethanesulfonyl) - β -D-glucopyranosyl } -2-deoxy-2- (1, 3-dioxo-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (the compound represented by formula C-11 (wherein X ₁ is Tf group)) was obtained as an ethyl acetate solution. The product was used directly in the next step.

Example 18

4-Methoxyphenyl 3, 6-di-O-benzyl-4-O- {4, 6-O-benzylidene-3-O- [ (naphthalen-2-yl) methyl ] -beta-D-mannopyranosyl } -2- (2-carboxybenzamide) -2-deoxy-beta-D-glucopyranoside (a compound represented by formula C-13)

To a solution (90 mL) of the 4-methoxyphenyl 3, 6-di-O-benzyl-4-O- {4, 6-O-benzylidene-3-O- [ (naphthalen-2-yl) methyl ] -2-O- (trifluoromethanesulfonyl) - β -D-glucopyranosyl } -2-deoxy-2- (1, 3-di-oxo-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (compound represented by formula C-11 (wherein X ₁ is Tf group)) obtained in example 17, dimethyl sulfoxide (150 mL) and tetrabutylammonium acetate (17.67 g,58.59 mmol) were added, and after heating to 30℃and stirring for 17 hours, the end of the reaction was confirmed by HPLC. Toluene (150 mL) was added to the reaction mixture, and the mixture was concentrated under reduced pressure until the amount of the solution became 165mL. Methanol (45 mL) and 50% aqueous sodium hydroxide (3.52 g,87.88 mmol) were added and stirred at 25℃for 1.5 hours. After completion of the reaction was confirmed by HPLC, ethyl acetate (450 mL) and water (300 mL) were added to separate the solution. Water (300 mL) was added to the organic layer, cooled to 0deg.C, and the pH was adjusted to 2.73 with 6N hydrochloric acid under vigorous stirring. Tetrahydrofuran (300 mL) was added to the separated organic layer, and the mixture was concentrated under reduced pressure until the liquid volume became 150mL. Tetrahydrofuran (300 mL) was added thereto, and the mixture was concentrated again until the liquid volume became 90mL, and the temperature was adjusted to 45℃at an internal temperature. After tetrahydrofuran (60 mL) was added, the mixture was cooled to 25℃and 2-propanol (150 mL) and water (15 mL) were added, and 4-methoxyphenyl 3, 6-di-O-benzyl-4-O- {4, 6-O-benzylidene-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl } -2- (2-carboxybenzamide) -2-deoxy- β -D-glucopyranoside (compound represented by formula C-13) (30 mg) was added. After stirring at 25℃for 14 hours and confirming the precipitation of crystals, 2-propanol (210 mL) was added dropwise over 1 hour and cooled to 0 ℃. After stirring for 2 hours, the precipitated crystals were filtered off and washed with 2-propanol (150 mL) cooled to 0 ℃. The obtained crystals were dried under reduced pressure at 40℃to give 4-methoxyphenyl 3, 6-di-O-benzyl-4-O- {4, 6-O-benzylidene-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl } -2- (2-carboxybenzamide) -2-deoxy- β -D-glucopyranoside (compound represented by formula C-13) (27.74 g, yield 94.3%, based on the compound represented by formula C-8).

The process of the compound represented by the formula C-12 to the compound represented by the formula C-13 is carried out by a one-pot method.

As the seed crystal, a seed crystal is used, which is obtained by separating a reaction solution into small portions and concentrating the reaction solution to precipitate a solid.

For the reaction, tetrabutylammonium acetate (product code: 86849, purity: > 90%) manufactured by tokyo chemical industry Co., ltd. (product code: T2694, purity: > 90.0%) and Sigma-Aldrich Co., ltd.) were used to perform the reaction well. In some cases, tetrabutylammonium acetate may contain an excessive amount of acetic acid in other factories, and in this case, the reaction tends to be significantly delayed. As an alternative method, the same conversion reaction can be performed using cesium acetate (details are described below).

The conversion of the compound represented by the formula C-11 to the compound represented by the formula C-12 can be carried out under conditions of cesium acetate (3 equivalents), dimethyl sulfoxide, 50 ℃ and 24 hours. Thereafter, the compound represented by the formula C-13 can be obtained by performing the same reaction (compound represented by the formula C-12. Fwdarw. Compound represented by the formula C-13) and post-treatment.

¹H－NMR(500MHz,CDCl₃)δ8.02(dd,J＝6.0,2.0Hz,1H),7.69－7.83(m,4H),7.38－7.49(m,12H),7.32－7.34(m,2H),7.16－7.29(m,8H),6.97(ddd,J＝9.0,4.0,2.5Hz,2H),6.76(ddd,J＝9.5,3.5,2.5Hz,2H),5.51(s,1H),5.40(d,J＝6.0Hz,1H),4.83－4.91(m,3H),4.76(d,J＝11.5Hz,1H),4.55(d,J＝0.5Hz,1H),4.49(d,J＝12.0Hz,1H),4.36(d,J＝12.0Hz,1H),4.26－4.30(m,1H),4.16(t,J＝6.5Hz,1H),4.05－4.09(m,2H),3.99(dd,J＝3.0,0.5Hz,1H),3.93(t,J＝9.5Hz,1H),3.80－3.86(m,2H),3.71(s,3H),3.65－3.69(m,1H),3.56(t,J＝10.0Hz,1H),3.51(dd,J＝10.0,3.5Hz,1H),3.13(td,J＝9.5,5.0Hz,1H).

¹³C－NMR(125MHz,CDCl₃)δ170.9,168.4,155.3,151.4,138.7,138.0,137.6,136.2,135.4,133.4,133.3.,132.2,132.1,130.7,130.3,129.2,128.6,128.49,128.45,128.11,128.07,128.0,127.89,127.87,127.8,126.8,126.4,126.3,126.2,125.8,118.6,114.7,101.7,100.4,99.1,78.3,76.7,76.4,75.1,73.7,73.3,72.5,69.9,69.4,68.5,67.0,55.8,54.4.

HRMS (ESI ⁺)[M+H]⁺C₅₉H₅₈NO₁₄ theory: 1004.3852; experimental 1004.3873).

Example 19

4-Methoxyphenyl 3, 6-di-O-benzyl-4-O- {4, 6-O-benzylidene-3-O- [ (naphthalen-2-yl) methyl ] -beta-D-mannopyranosyl } -2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranoside (a compound represented by formula C-14)

To a solution of 4-methoxyphenyl 3, 6-di-O-benzyl-4-O- {4, 6-O-benzylidene-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl } -2- (2-carboxybenzamide) -2-deoxy- β -D-glucopyranoside (compound of formula C-13) (6.00 g,5.98 mmol) in methylene chloride (30 mL) was added 1-hydroxybenzotriazole monohydrate (0.18 g,1.20 mmol), N-diisopropylethylamine (0.85 g,6.57 mmol) and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (1.26 g,6.57 mmol), and the mixture was stirred for 31 hours at 40 ℃. After confirming completion of the reaction by HPLC, the reaction solution was cooled to 0℃and then ethyl acetate (90 mL) and water (60 mL) were added thereto, and 6N hydrochloric acid was added thereto with vigorous stirring until the pH became 7. After separating the organic layer and the aqueous layer, the organic layer was washed with water (60 mL) and saturated brine (30 mL). The solution was concentrated under reduced pressure to a liquid volume of 12mL. Tetrahydrofuran (60 mL) was added and concentrated again to a liquid volume of 9mL to give a tetrahydrofuran solution of the crude product 4-methoxyphenyl 3, 6-di-O-benzyl-4-O- {4, 6-O-benzylidene-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl } -2-deoxy-2- (1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (compound of formula C-14). The product was used directly in the next step.

Example 20

4-Methoxyphenyl 3, 6-di-O-benzyl-4-O- { 2-O-benzyl-4, 6-O-benzylidene-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl } -2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (a compound represented by formula C-15)

To a solution of the crude product 4-methoxyphenyl 3, 6-di-O-benzyl-4-O- {4, 6-O-benzylidene-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl } -2-deoxy-2- (1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (compound represented by formula C-14) was added N, N-dimethylacetamide (60 mL), benzyl bromide (1.53 g,8.96 mmol), methyl trifluoroacetate (0.15 g,1.20 mmol) and molecular sieve 4A (1.8 g), and a solution of lithium t-butoxide in tetrahydrofuran (note: the present solution was stirred for 30 minutes by cooling a mixed solution of 2-methyl-2-propanol (0.66 g,8.96 mmol) and tetrahydrofuran (2.4 mL) to 0℃and adding a hexane solution (1.55 mol/L) (5.78 mmol, 8.96 mmol) of N-butyllithium). After completion of the reaction, ethylenediamine (anhydrous) (0.18 g,2.99 mmol) was added thereto and the mixture was stirred for 1 hour. To the solution was added acetic acid (0.72 g,11.95 mmol), and the mixture was filtered. After washing molecular sieve 4A with ethyl acetate (90 mL), water (60 mL) was added for separation. After washing the organic layer twice with water (60 mL), it was concentrated under reduced pressure to a liquid volume of 12mL. Toluene (30 mL) was added thereto and the mixture was concentrated again until the liquid volume became 12mL, toluene (18 mL) and silica gel 60N (spherical, manufactured by Kanto chemical Co., ltd., particle size: 40 to 50 μm) (9 g) were added thereto, and the mixture was stirred at 25℃for 30 minutes. The suspension was filtered and the silica gel was washed with a mixed solution of toluene (191 mL) and ethyl acetate (19 mL). The filtrate was concentrated under reduced pressure to a liquid volume of 9mL to give a toluene solution of the crude product 4-methoxyphenyl 3, 6-di-O-benzyl-4-O- { 2-O-benzyl-4, 6-O-benzylidene-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl } -2-deoxy-2- (1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (a compound represented by formula C-15).

Example 21

4-Methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-4-O- {2, 4-di-O-benzyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl } -2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (a compound represented by formula A-4)

To the resulting crude product, a solution of 4-methoxyphenyl 3, 6-di-O-benzyl-4-O- { 2-O-benzyl-4, 6-O-benzylidene-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl } -2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (compound represented by formula C-15) (5.50 g (5.48 mmol) based on the compound represented by formula C-14) was added dichloromethane (16.5 mL) and molecular sieve 4A (550 mg) cooled to 0 ℃, borane-tetrahydrofuran complex (0.91 mol/L tetrahydrofuran solution) (18.06 mL,16.43 mmol) and copper (II) trifluoroacetate (0.59 g,1.64 mmol) were added and stirred for 3 hours. After confirming the completion of the reaction by HPLC, methanol (5.5 mL) was added and further stirred for 30 minutes. The solution was filtered, and after washing molecular sieve 4A with ethyl acetate (110 mL), 0.5N hydrochloric acid (55 mL) was added and stirred for 30 minutes. After separating the organic layer and the aqueous layer, the organic layer was washed with 0.5N hydrochloric acid (55 mL) and saturated brine (27.5 mL), and the solution was concentrated under reduced pressure to dryness. The product was purified by silica gel column chromatography (silica gel 300g, hexane: ethyl acetate=55:45→30:70) to give 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-4-O- {2, 4-di-O-benzyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl } -2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (compound represented by formula a-4) (4.94 g, yield 83.6%, HPLC area: 98.54%).

¹H－NMR(500MHz,CDCl₃)δ7.67－7.84(m,8H),7.44－7.49(m,4H),7.40(dd,J＝8.0,1.5Hz,1H),7.21－7.33(m,13H),6.87－6.93(m,5H),6.82(ddd,J＝9.5,4.0,2.5Hz,2H),6.70(ddd,J＝9.0,4.0,2.0Hz,2H),5.64(d,J＝8.5Hz,1H),4.94(d,J＝12.5Hz,1H),4.91(d,J＝10.0Hz,1H),4.90(s,2H),4.66(s,2H),4.60(d,J＝11.0Hz,1H),4.59(d,J＝12.0Hz,1H),4.55(s,1H),4.40－4.46(m,3H),4.33(dd,J＝11.0,9.0Hz,1H),4.06(dd,J＝9.5,8.5Hz,1H),3.80－3.85(m,2H),3.70－3.76(m,2H),3.71(s,3H),3.61－3.68(m,2H),3.45－3.48(m,1H),3.44(dd,J＝9.5,3.0Hz,1H),3.23(ddd,J＝9.5,5.5,2.5Hz,1H),1.97(br－t,1H).

¹³C－NMR(125MHz,CDCl₃)δ155.6,151.1,138.9,138.64,138.56,138.0,135.9,134.0,133.5,133.2,131.8,128.7,128.6,128.4,128.3,128.20,128.19,128.15,128.1,120.04,128.01,127.9,127.7,127.6,127.3,126.4,126.3,126.1,125.8,123.6,119.0,114.6,101.2,98.0,82.6,79.0,77.2,75.9,75.4,75.3,75.2,75.1,74.8,74.7,73.8,72.1,68.7,62.6,55.82,55.78,34.4,30.5.

HRMS (ESI ^－)[M+HCO₂]^－C₆₇H₆₄NO₁₅ theory: 1122.4281; experimental 1122.4285).

Example 22

A toluene solution (corresponding to 78.4 mmol) of 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-4-O- {2, 4-di-O-benzyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl } -2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (compound represented by formula A-4) (65.0 g,60.3 mmol) and 2-O-acetyl-3, 4, 6-tri-O-benzyl-1-O- (2, 2-trichloroethaneimido) -D-mannopyranose (compound represented by formula A-3) was added to a 1L four-necked flask, which was dissolved in 650mL of toluene, and molecular sieve 4A powder (10 μm or less, 13.0 g) was added. Trimethylsilicone triflate (2.7 mL,15.1 mmol) was added dropwise under nitrogen at-15℃over 15 minutes and stirred at the same temperature for 30 minutes. After completion of the reaction by HPLC, triethylamine (4.2 mL,30.2 mmol) was added and the mixture was warmed to room temperature. The reaction mixture was filtered through celite, and then washed with acetonitrile (195 mL). The filtrate was concentrated under reduced pressure, acetonitrile (650 mL) was added to the concentrated residue, and silica gel 120RP-18 (manufactured by Kanto chemical Co., ltd., particle size: 40 to 50 μm,97.5 g) for reversed phase was added. Water (130 mL) was added dropwise over 30 minutes to adsorb the target onto the solid phase, followed by filtration. After washing the solid phase with acetonitrile/water (3/1, 326 mL) (filtrate discard), the target was desorbed with acetonitrile (585 mL) -ethyl acetate (65 mL) solution. The filtrate was concentrated under reduced pressure, whereby 4-methoxyphenyl 2-O-acetyl-3, 4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1→6) -2, 4-di-O-benzyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl- (1→4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-bridged oxy-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (compound represented by formula A-5) (70.5 g, isolated yield 94%) was obtained in the form of white amorphous.

¹H－NMR(CDCl₃)σ7.85－7.80(m,1H),7.78(d,J＝8.6Hz,1H),7.74－7.58(m,5H),7.50－7.43(m,5H),7.39(dd,J＝8.6,1.7Hz,1H),7.32－7.02(m,30H),6.92－6.89(m,2H),6.78－6.76(m,2H),6.71－6.62(m,5H),5.59(d,J＝8.6Hz,1H),5.36(dd,J＝2.9,2.3Hz,1H),5.00－4.90(m,4H),4.87(d,J＝12.6Hz,1H),4.79(d,J＝11.5Hz,1H),4.66－4.50(m,7H),4.43(dd,J＝11.5,2.3Hz,1H),4.40－4.34(m,3H),4.28(dd,J＝10.3,8.6Hz,1H),4.22(d,J＝11.5Hz,1H),4.08(dd,J＝9.7,9.2Hz,1H),3.94(dd,J＝9.2,9.2Hz,1H),3.89－3.83(m,3H),3.83－3.58(m,10H),3.55－3.50(m,1H),3.42(dd,J＝9.2,2.9Hz,1H),3.37－3.31(m,1H),1.90(s,3H).

¹³C－NMR(CDCl₃)σ167.0,155.3,150.8,138.8,138.7,138.51,138.49,138.4,138.0,137.9,135.6,133.6,133.2,132.9,131.6,128.44,128.41,128.32,128.26,128.22,128.19,128.12,127.86,127.81,127.75,127.71,127.69,127.63,127.56,127.44,127.37,127.27,127.0,126.3,126.2,125.9,125.7,123.2,118.7,114.3,101.9,98.3,97.6,82.7,79.6,77.8,76.6,75.04,74.98,74.91,74.89,74.75,74.4,74.3,74.1,74.0,73.4,73.3,71.70,71.65,71.3,68.8,68.6,68.3,67.1,55.6,55.5,20.9.

HRMS (ESI ⁺)[M+HNEt₃]⁺C₁₀₁H₁₀₉N₂O₁₉ ⁺ theory: 1654.7653; experimental 1654.7618).

[α]_D ²⁰＝+31.589(c 1.002,CDCl₃)。

Example 23

4-Methoxyphenyl 2-O-acetyl-3, 4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 6) -2, 4-di-O-benzyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (a compound of formula A-5) (44.0 g,28.3 mmol) is added to a 1L four-necked flask, and dichloromethane (228 mL), 1, 3-hexafluoro-2-propanol (163 mL) and water (14 mL) are added. A solution of [ bis (trifluoroacetoxy) iodo ] benzene (25.9 g,56.6 mmol) in dichloromethane (80 mL) and trifluoroacetic acid (6.9 mL,84.9 mmol) were added under nitrogen at 2deg.C and stirred at the same temperature for 8 hours. After confirming completion of the reaction by HPLC, a sodium hydrogencarbonate (18.7 g) -water (228 mL) solution was added and stirred for 5 minutes, and a sodium sulfite (11.7 g) -water (228 mL) solution was added. After stirring for 5 minutes, the mixture was allowed to stand, and the dichloromethane layer was separated. After concentrating the obtained organic layer under reduced pressure to 97.5mL, ethyl acetate (325 mL) and sodium chloride (23.4 g) -water (211 mL) were added, respectively. After stirring for 5 minutes, the mixture was allowed to stand, and the organic layer was separated. After the obtained organic layer was concentrated under reduced pressure to 97.5mL, toluene (890 mL) was added and concentrated under reduced pressure again to 97.5mL. Toluene (164 mL) and methylene chloride (65 mL) were added, and silica gel 60N (particle size: 40 to 50 μm,130g, manufactured by Kanto chemical Co., ltd.) was added to adsorb the target substance on silica gel, followed by filtration. The reaction mixture was washed with a solution of dichloromethane (130 mL) -toluene (520 mL) (filtrate was discarded), and the target was desorbed from the solid phase with a solution of ethyl acetate (220 mL) -dichloromethane (455 mL). After concentrating the filtrate under reduced pressure, toluene (228 mL) was added and concentrated under reduced pressure to 97.5mL, whereby 2-O-acetyl-3, 4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 6) -2, 4-di-O-benzyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -D-glucopyranose (a compound represented by formula A-6) was obtained as a toluene solution. The solution was used directly in the next step.

¹H－NMR(CDCl₃)σ7.85－7.80(m,1H),7.78(d,J＝8.6Hz,1H),7.72(s,1H),7.70－7.64(m,1H),7.63(brs,2H),7.48－7.43(m,4H),7.39(dd,J＝8.6,1.7Hz,1H),7.33－7.10(m,30H),6.92－6.87(m,2H),6.77－6.66(m,3H),5.38(dd,J＝3.4,1.7Hz,1H),5.23(dd,J＝8.6,8.6Hz,1H),4.97－4.86(m,4H),4.84(d,J＝12.6Hz,1H),4.79(d,J＝10.9Hz,1H),4.63(d,J＝11.5Hz,1H),4.60－4.48(m,7H),4.46(d,J＝11.5Hz,1H),4.37(dd,J＝12.0,3.4Hz,1H),4.32(dd,J＝10.9,8.6Hz,1H),4.23(d,J＝10.9Hz,1H),4.06(dd,J＝9.2,9.2Hz,1H),4.01(dd,J＝10.9,9.2Hz,1H),3.92(dd,J＝9.2,3.4Hz,1H),3.89－3.74(m,6H),3.72－3.65(m,2H),3.64－3.57(m,2H),3.48(dd,J＝10.9,1.1Hz,1H),3.42－3.34(m,2H),2.68(dd,J＝9.2,1.1Hz,1H),1.95(s,3H).

¹³C－NMR(CDCl₃)σ170.2,167.9,138.85,138.79,138.6,138.5,138.4,138.0,137.8,135.6,133.6,133.2,132.9,131.6,128.55,128.50,128.33,128.24,128.20,128.16,127.9,127.80,127.75,127.69,127.66,127.62,127.54,127.45,127.36,127.33,126.97,126.22,126.17,125.9,125.6,123.2,101.1,97.9,92.9,82.6,78.6,78.0,76.1,74.9,74.8,74.4,74.15,74.13,74.0,73.5,73.2,71.7,71.6,71.3,68.8,68.768.4,67.0,57.6,20.9.

HRMS (ESI ⁺)[M+HNEt₃]⁺C₉₄H₁₀₃N₂O₁₈ ⁺ theory: 1548.7234; experimental 1548.7237).

[α]_D ²⁰＝+32.528(c 1.006,CDCl₃)。

Example 24

A toluene solution of 2-O-acetyl-3, 4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 6) -2, 4-di-O-benzyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -D-glucopyranose (a compound represented by formula A-6) obtained in example 23 was charged into a 1L four-necked flask, methylene chloride (260 mL) and a powder of molecular sieve 4A (10 μm or less, 21.8 g) were added, and cooled to 0 ℃.1, 8-diazabicyclo [5.4.0] undec-7-ene (5.08 mL,34.0 mmol) and 2, 2-trifluoro-N-phenyliminoacetyl chloride (5.25 mL,31.1 mmol) were added under nitrogen at the same temperature and stirred for 5 hours. The reaction solution was filtered using a neutral silica gel pad (silica gel 60N, manufactured by Kanto chemical Co., ltd., particle size: 40 to 50 μm,130 g) filled with methylene chloride. The silica gel pad was washed with 10% ethyl acetate/methylene chloride (1760 mL, 220mL each) and the main fraction was concentrated under reduced pressure, followed by addition of toluene (228 mL) and concentration under reduced pressure to 97.5mL, whereby a toluene solution of 2-O-acetyl-3, 4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1.fwdarw.6) -2, 4-di-O-benzyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dimethoxy-1, 3-dihydro-2H-isoindol-2-yl) -1-O- (2, 2-trifluoro-N-phenylethanimido) -D-glucopyranose (a compound represented by formula A-7) was obtained. The solution was used directly in the next step.

¹H－NMR(CDCl₃)σ7.85－7.80(m,1H),7.78(d,J＝8.6Hz,1H),7.72(s,1H),7.70－7.64(m,3H),7.50－7.43(m,4H),7.39(dd,J＝8.6,1.7Hz,1H),7.33－7.08(m,30H),7.06－7.01(m,1H),6.88(d,J＝6.9Hz,2H),6.70－6.60(m,4H),5.35(dd,J＝2.9,1.7Hz,1H),4.97－4.88(m,4H),4.84(d,J＝13.2Hz,1H),4.79(d,J＝10.9Hz,1H),4.65－4.49(m,7H),4.43(dd,J＝11.5,4.0Hz,1H),4.36(dd,J＝12.0,7.4Hz,1H),4.22(d,J＝11.5Hz,1H),4.11－4.04(m,1H),3.93(dd,J＝9.7,9.7Hz,1H),3.89－3.83(m,2H),3.82－3.69(m,4H),3.64(dd,J＝10.9,4.0Hz,1H),3.52(dd,J＝10.9,1.1Hz,1H),3.39(dd,J＝9.7,2.9Hz,1H),3.33－3.28(m,1H),1.89(s,3H).

¹³C－NMR(CDCl₃)σ170.0,167.4,143.0,138.8,138.7,138.5,138.4,138.0,137.6,135.6,133.7,133.2,133.0,131.5,128.56,128.53,128.4,128.32.128.30,128.26,128.22,128.19,128.14,128.12,127.90,127.86,127.81,127.74,127.72,127.69,127.54,127.46,127.38,127.28,127.0,126.3,126.2,126.0,125.6,124.3,123.3,119.3,101.8,98.2,82.6,79.0,77.8,76.2,75.5,75.0,74.91,74.89,74.7,74.5,74.3,74.1,74.0,73.4,73.3,71.7,71.7,71.3,68.8,68.3,68.0,67.0,54.7,20.8.

HRMS(ESI⁺)[M+HNH₃]⁺C₉₆H₉₁F₃N₃O₁₈ ⁺ Theoretical value: 1635.6591; experimental values 1635.6549.

[α]_D ²⁰＝+62.169(c 1.002,CDCl₃)。

Example 25

A toluene solution of 2-O-acetyl-3, 4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 6) -2, 4-di-O-benzyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -1-O- (2, 2-trifluoro-N-phenylethanimido) -D-glucopyranose (compound of formula A-7), toluene (488 mL) and 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (compound of formula A-8) (21.55 g, 36.6 g) obtained in example 24 were added to a four-necked flask (2 mL) and the mixture was stirred into a molecular sieve (14.55 g). Trimethylsilicone triflate (545. Mu.L, 2.83 mmol) was added dropwise under nitrogen at-15℃over 5 minutes and stirred at the same temperature for 1 hour. After completion of the reaction by HPLC, triethylamine (1.67 mL,11.32 mmol) was added and the mixture was warmed to room temperature. The reaction solution was filtered and washed with acetonitrile (160 mL). The filtrate was concentrated under reduced pressure to 97.5mL, acetonitrile (650 mL) was added and concentrated again under reduced pressure to 97.5mL. Acetonitrile (488 mL) and silica gel 120RP-18 (manufactured by Kanto chemical Co., ltd., particle size: 40 to 50 μm,130 g) for reversed phase were added. Water (146 mL) was added dropwise over 30 minutes to adsorb the target onto the solid phase, followed by filtration. The target was desorbed by washing with acetonitrile (536 mL) -water (146 mL) (filtrate discard) and acetonitrile (975 mL) -ethyl acetate (244 mL). The filtrate was concentrated under reduced pressure, and the resulting solution was azeotroped twice with toluene (325 mL) to give a final solution of 97.5mL, whereby 4-methoxyphenyl 2-O-acetyl-3, 4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 6) -2, 4-di-O-benzyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (a compound represented by formula A-9) was obtained as a toluene solution. A part (90 mL, 26.1mmol based on the compound represented by formula A-4) of the present solution was collected and used in the next step.

¹H－NMR(CDCl₃)σ7.86－7.81(m,1H),7.81－7.75(m,2H),7.74－7.60(m,7H),7.56－7.44(m,5H),7.40(dd,J＝8.6,1.1Hz,1H),7.28－7.05(m,35H),6.95－6.91(m,2H),6.91－6.88(m,2H),6.78－6.62(m,8H),6.61－6.55(m,2H),5.42(d,J＝8.6Hz,1H),5.33(dd,J＝2.9,1.7Hz,1H),5.23(d,J＝8.0Hz,1H),4.98(d,J＝12.6Hz,1H),4.94－4.85(m,4H),4.82(d,J＝13.2Hz,1H),4.76(d,J＝10.9Hz,1H),4.65－4.29(m,16H),4.25－4.12(m,5H),4.04(dd,J＝10.3,8.0Hz,1H),3.95(dd,J＝9.2,9.2Hz,1H),3.89－3.80(m,3H),3.77(d,J＝9.2Hz,1H),3.73(d,J＝10.9Hz,1H),3.70－3.65(m,1H),3.64(s,3H),3.63－3.57(m,2H),3.53－3.46(m,2H),3.44－3.35(m,4H),3.30－3.23(m,2H),1.83(s,3H).

¹³C－NMR(CDCl₃)σ169.8,168.1,167.4,155.2,150.8,138.9,138.71,138.68,138.55,138.48,138.36,137.9,135.6,133.8,133.6,133.2,133.0,131.8,131.6,131.5,128.5,128.34,128.32,128.29,128.26,128.20,128.17,128.12,128.10,127.89,127.85,127.80,127.77,127.69,127.65,127.52,127.47,127.44,127.36,127.29,127.28,126.90,126.87,126.3,126.2,126.0,125.7,123.5,123.2,123.1,118.5,114.2,102.1,98.2,97.4,97.1,82.7,80.0,77.8,76.6,75.9,74.9,74.8,74.63,74.61,74.56,74.49,74.3,74.06,74.03,73.3,73.2,72.6,71.7,71.6,71.2,68.7,68.3,68.11,68.06,68.8,56.5,55.6,55.5,20.8.

HRMS (ESI ⁺)[M+HNEt₃]⁺C₁₂₉H₁₃₄N₃O₂₅ ⁺ theory: 2125.9334; experimental 2125.9267).

[α]_D ²⁰＝+30.098(c 1.008,CDCl₃)。

Example 26

A toluene solution (90 mL) of 4-methoxyphenyl 2-O-acetyl-3, 4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 6) -2, 4-di-O-benzyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-bridged oxy-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-bridged oxy-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (compound represented by formula A-9), 26.1mmol based on the compound represented by formula A-4, was added to a 1L four-necked flask, and tetrahydrofuran (165.8 mL) and three-necked acetic acid (165.27 mL) was added. After stirring at room temperature under nitrogen for 5 minutes, a 1M solution of potassium tert-butoxide in tetrahydrofuran (13.9 mL,13.9 mmol) was added and stirred at 40℃for 1 hour. After completion of the reaction, the reaction mixture was cooled to room temperature by HPLC. After acetic acid (1.11 mL,19.4 mmol) was added, ethyl acetate (330 mL) and water (270 mL) were added. After triethylamine was added until the pH became 7, sodium chloride (2.7 g) was added and stirred for 5 minutes. After standing, the resulting organic layer was separated, and washed twice with water (270 mL). The organic layer was concentrated to 90mL under reduced pressure, toluene (300 mL) was added thereto and concentrated again to 90mL under reduced pressure, whereby 4-methoxyphenyl 3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 6) -2, 4-di-O-benzyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (a compound represented by formula A-10) was obtained as a toluene solution. The solution was used directly in the next step.

¹H－NMR(CDCl₃)σ7.85－7.81(m,1H),7.79(d,J＝8.6Hz,1H),7.78－7.72(m,2H),7.72－7.60(m,6H),7.59－7.54(m,1H),7.50－7.44(m,4H),7.41(dd,J＝8.6,1.1Hz,1H),7.31－7.13(m,32H),7.10－7.05(m,2H),6.97－6.93(m,2H),6.90－6.86(m,2H),6.78－6.67(m,8H),6.61－6.56(m,2H),5.43(d,J＝8.6Hz,1H),5.25(d,J＝8.0Hz,1H),4.99－4.86(m,4H),4.83(d,J＝12.6Hz,1H),4.71(d,J＝10.9Hz,1H),4.64(d,J＝12.0Hz,1H),4.61(d,J＝12.0Hz,1H),4.56(d,J＝6.9Hz,1H),4.53－4.39(m,7H),4.39－4.31(m,5H),4.30(d,J＝11.5Hz,1H),4.25－4.14(m,4H),4.05(dd,J＝9.7,8.6Hz,1H),3.94－3.88(m,2H),3.86－3.80(m,2H),3.76－3.68(m,3H),3.68－3.59(m,5H),3.57－3.35(m,7H),3.31－3.25(m,2H),2.15(d,J＝4.0Hz,1H).

¹³C－NMR(CDCl₃)σ168.2,167.5,155.2,150.8,138.91,138.88,138.6,138.52,138.47,138.38,128.32,128.0,137.9,135.7,133.9,133.6,133.3,133.0,131.8,131.6,131.4,128.5,128.29,128.23,128.19,128.13,128.0,127.92,127.87,127.82,127.79,127.76,127.70,127.56,127.53,127.46,127.36,127.31,127.28,126.9,126.3,126.2,126.0,125.7,123.5,123.2,123.1,118.6,114.2,101.9,99.7,97.4,97.1,82.7,79.7,79.6,76.7,75.9,75.3,74.9,74.8,74.73,74.68,74.63,74.5,74.4,74.2,74.1,73.2,73.2,72.6,71.8,71.3,71.2,68.9,68.13,68.07,67.8,66.6,56.5,55.6,55.5.

HRMS (ESI ⁺)[M+HNEt₃]⁺C₁₂₇H₁₃₂N₃O₂₄ ⁺ theory: 2083.9229; experimental 2083.9150).

[α]_D ²⁰＝+27.776(c 1.007,CDCl₃)。

Example 27

In a toluene solution of 4-methoxyphenyl 3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 6) -2, 4-di-O-benzyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (a compound represented by formula A-10) obtained in example 26, A toluene solution (118.2 g,36.2 mmol) of methylene chloride (450 mL) and 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -4-O- (2, 3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl) -1-O- (2, 2-trifluoro-N-phenylethaneimido) -beta-D-glucopyranose (a compound represented by the formula A-11) was charged into a 1L four-necked flask, and a molecular sieve 4A powder (10 μm or less, 15.0 g) was added. T-butyldimethylsilyl triflate (2.4 mL,10.4 mmol) was added dropwise under nitrogen at-78deg.C over 5min and stirred at the same temperature for 10 h. Triethylamine (4.6 mL,33.2 mmol) was added and the temperature was raised at room temperature. The reaction mixture was filtered through celite and washed with acetonitrile (150 mL). The filtrate was concentrated to 150mL under reduced pressure, acetonitrile (600 mL) was added thereto, and then concentrated to 150mL under reduced pressure. Acetonitrile (600 mL) and silica gel 120RP-18 for reversed phase (manufactured by Kanto chemical Co., ltd., particle diameter: 40 to 50 μm,135 g) were added. Water (120 mL) was added dropwise over 30 minutes to adsorb the target onto the solid phase, followed by filtration. After washing the solid phase with acetonitrile (900 mL) -water (135 mL) (washing waste), the target was desorbed from the solid phase with acetonitrile (840 mL) -ethyl acetate (210 mL) solution. After concentrating the desorption solution under reduced pressure, toluene (300 mL) was added thereto and concentrated under reduced pressure to 90mL, whereby 4-methoxyphenyl 2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-bridged oxy-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1.fwdarw.2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1.fwdarw.6) -2, 4-di-O-benzyl-3-O- [ (naphthalen-2-yl) methyl ] -beta-D-mannopyranosyl- (1.fwdarw.4) -3, Toluene solution of 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranoside (compound represented by formula A-12). The solution was used directly in the next step.

¹H－NMR(CDCl₃)σ7.84－7.73(m,4H),7.70－7.57(7H),7.55－7.03(m,68H),6.99－6.89(m,11H),6.88－6.83(m,1H),6.82－6.67(m,7H),6.67－6.62(m,2H),6.62－6.56(m,3H),5.42(d,J＝8.6Hz,1H),5.23(d,J＝8.6Hz,1H),5.02(d,J＝8.6Hz,1H),4.95－4.89(m,3H),4.86－4.79(m,5H),4.77－4.66(m,4H),4.64－4.08(m,30H),4.06－3.98(m,3H),3.91－3.68(m,9H),3.65(s,3H),3.58(d,J＝Hz,1H),3.54－3.32(m,13H),3.32－3.27(m,1H),3.22－3.16(m,3H),2.84(dd,J＝10.9,5.7Hz,1H).

¹³C－NMR(CDCl₃)σ168.4,168.2,167.4,155.2,150.8,139.1,139.02,139.99,138.93,138.8,138.7,138.52,138.48,138.38,138.32,138.06,138.01,135.6,133.7,133.61.133.56,133.3,133.2,132.9,131.8,131.6,131.4,128.54,128.50,128.39,128.34,128.17,128.13,128.10,128.00,127.96,127.89,127.82,127.78,127.67,127.64,127.62,127.59,127.51,127.48,127.46,127.41,127.39,127.29,127.20,127.05,127.01,126.9,126.7,126.3,126.2,125.9,125.7,123.5,123.2,123.1,123.0,118.6,114.2,102.9,102.7,97.8,97.4,97.0,96.9,83.1,82.5,80.4,79.9,77.7,77.4,76.9,76.6,75.7,75.3,75.175.0,74.8,74.7,74.6,74.52,74.46,74.0,73.95,73.85,73.7,73.4,73.2,73.0,72.9,72.58,72.55,72.52,72.48,72.0,71.9,69.8,69.7,68.2,3,68.20,68.06,68.03,67.0,56.6,55.6,55.5.

HRMS (ESI ⁺)[M+HNEt₃]⁺C₁₉₁H₁₇₅N₄O₃₅ ⁺ theory: 3078.3350; experimental 3078.3200).

[α]_D ²⁰＝+9.276(c 1.002,CDCl₃)。

Example 28-1

4-Methoxyphenyl 2,3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl- (1→4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranosyl- (1→2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1→6) -2, 4-di-O-benzyl-3-O- [ (naphthalen-2-yl) methyl ] - β -D-mannopyranosyl- (1→4) -3, 6-di-O-benzyl-2-deoxy-2- (1), A toluene solution (equivalent to 64.2 mmol) of 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranoside (a compound represented by formula A-12) is introduced into a 3L four-necked flask, and 1, 3-hexafluoro-2-propanol (1337 mL) and water (133.7 mL) are added. After cooling the reaction solution to about-30 ℃,2, 3-dichloro-5, 6-dicyano-p-benzoquinone (17.48 g,77.0 mmol) was added and stirred for 38 hours. After confirming that the reaction was sufficiently progressed, a solution of sodium sulfite (4.04 g,32.1 mmol) -water (95.6 mL) was added to stop the reaction, and the temperature was raised to room temperature over 1 hour. Dichloromethane (1910 mL) and sodium bicarbonate (38.2 g) -sodium sulfite (38.2 g) -water (1910 mL) solutions were added separately and stirred for 5 minutes. After standing, the organic layer was separated, and concentrated under reduced pressure to 573mL. Toluene (955 mL) was added to the residue, concentrated under reduced pressure to 573mL, and a solution of ethyl acetate (955 mL) and sodium chloride (95.5 g) -water (860 mL) was added and stirred for five minutes. After standing, the organic layer was concentrated under reduced pressure to 382mL, acetonitrile (1910 mL) was added thereto, and the mixture was concentrated under reduced pressure to 382mL again. Acetonitrile (1528 mL) and silica gel 120RP-18 (particle size 40-50 μm,573g, manufactured by Kanto chemical Co., ltd.) were added to the residue. Water (1242 mL) was added dropwise over 30 minutes to adsorb the target onto the solid phase, followed by filtration. After washing the solid phase with acetonitrile (1337 mL) -water (573 mL) (washing liquid waste), the solid phase was washed with methanol (955 mL). The target was then desorbed from the solid phase with a solution of acetonitrile (6876 mL) -tetrahydrofuran (764 mL). Concentrating the desorption solution under reduced pressure, drying to obtain 4-methoxyphenyl 2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-di-oxo-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 6) -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2- (1), 167g of 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranoside (compound of formula A-13).

By subjecting the obtained crude product to purification by preparative HPLC, 4-methoxyphenyl 2,3,4, 6-tetra-O-benzyl-. Beta. -D-galactopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-di-oxo-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1.fwdarw.2) -3,4, 6-tri-O-benzyl-. Alpha. -D-mannopyranosyl- (1.fwdarw.6) -2, 4-di-O-benzyl-. Beta. -D-mannopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-di-oxo-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1.4) -3, 6-di-O-benzyl-2- (1.fwdarw.6-di-O-benzyl-2-deoxy-2- (1.fwdarw.4) -beta. -D-glucopyranosyl- (1.6-di-oxo-2-isoindol-2-yl) was obtained in the form of a refined product (the general glycoside of the formula shown in the following Table 13).

¹H－NMR(CDCl₃)σ7.75(d,J＝8.0Hz,1H),7.69－7.40(10H),7.55－7.10(m,54H),7.06－6.97(5H),6.97－6.89(m,8H),6.88－6.84(m,1H),6.82－6.73(m,5H),6.72－6.67(m,2H),6.62－6.56(m,5H),5.42(d,J＝8.6Hz,1H),5.22(d,J＝8.0Hz,1H),5.03(d,J＝12.0Hz,1H),4.99(d,J＝8.6Hz,1H),4.92(d,J＝10.9Hz,1H),4.87－4.79(m,5H),4.75(d,J＝11.5Hz,1H),4.70(d,J＝12.0Hz,1H),4.67(d,J＝12.0Hz,1H),4.63(d,J＝6.3Hz,1H),4.61(d,J＝6.9Hz,1H),4.60－3.97(m,30H),3.88(d,J＝2.9Hz,1H),3.84(d,J＝12.0Hz,1H),3.82－3.67(m,5H),3.66－3.60(m,5H),3.54－3.33(m,14H),3.27－3.23(m,1H),3.20(d,J＝10.9Hz,1H),3.15－3.09(m,2H),2.83－2.78(m,1H),2.27－2.21(m,1H).

¹³C－NMR(CDCl₃)σ168.4,168.2,167.4,167.3,155.2,150.8,139.1,139.0,138.9,138.8,138.7,138.51,138.49,138.44,138.38,138.37,138.35,138.26,138.0,137.7,133.8,133.6,122.49,133.41,133.36,131.8,131.4,128.6,128.52,128.49,128.39,128.34,128.30,128.2,128.14,128.10,128.06,128.0,127.85,127.82,127.7,127.69,127.65,127.56,127.53,127.48,127.38,127.33,127.26,127.1,127.0,126.9,126.7,123.5,123.2,123.09,123.05,122.98,118.5,114.2,102.9,102.6,97.8,97.4,97.0,96.9,82.5,80.5,79.9,78.5,77.8,77.4,76.9,76.7,75.87,75.80,75.2,75.0,74.79,74.77,74.62,74.59,74.48,74.41,74.2,74.0,73.9,73.6,73.4,73.0,72.9,72.7,72.62,72.56,72.0,69.8,68.24,68.20,68.1,67.8,67.2,56.6,55.6,55.5,53.4.

HRMS (ESI ⁺)[M+HNEt₃]⁺C₁₇₂H₁₈₃N₄O₃₅ ⁺ theory: 2938.2725; experimental 2938.2516).

[α]_D ²⁰＝+14.385(c 1.005,CDCl₃)。

Example 28-2

The compound represented by formula A-13 was purified by using a method based on the following route. As shown below, by this purification method, the compound represented by formula A-13 can be obtained in high purity without HPLC separation and purification.

Synthesis of Methoxyphenyl 2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (2-carboxybenzamide) -beta-D-glucopyranosyl- (1.fwdarw.2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1.fwdarw.6) -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (2-carboxybenzamide) -beta-D-glucopyranosyl- (1.fwdarw.4) -3, 6-di-0-benzyl-2-deoxy-2- (2-carboxybenzamide) -beta-D-glucopyranoside tris (R) - (+) -1- (1-naphtalenyl) ethylamine salt (compound of formula A-14)

4-Methoxyphenyl 2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-di-oxo-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 6) -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-di-oxo-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt 4) -3, After 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranoside (a compound represented by formula A-13) (25.00 g,8.82mmol, purity 72.2 PA%) is dissolved in methylene chloride (250 mL), silica gel (62.5g,FUJI SILYSIA chromatography SMB 100-20/45) is subjected to a liquid passing, an elution operation is performed with diisopropyl ether/methylene chloride mixed solvent (7/93, 1000 mL) (fraction separation is performed every 125mL in the liquid passing and elution operation, Recovering the solution). The fractions were selected by performing purity measurement by HPLC, and the selected fractions were mixed and concentrated to 38mL. Tetrahydrofuran (125 mL) was added to the concentrated solution, and after concentrating to 38mL, tetrahydrofuran (100 mL), methanol (38 mL) and aqueous sodium hydroxide solution (4M, 16.5mL,7.5 eq) were added dropwise, and the mixture was stirred at 45℃for 1 hour. After completion of the reaction, the reaction mixture was cooled to 0℃and hydrochloric acid (6M, 11mL,7.5 eq.) was added dropwise thereto at 10℃or lower to neutralize the reaction mixture. Ethyl acetate (250 mL) and 3% brine (250 mL) were added, and the pH of the aqueous layer was adjusted to 2.0 or less with hydrochloric acid (6M) under stirring. The aqueous layer was removed by pipetting and the resulting organic layer was washed with 3% brine (250 mL). The organic layer was concentrated to 38mL, ethyl acetate (250 mL) was added and concentrated to 38mL. To the concentrate were added ethyl acetate (138 mL), (R) - (+) -1- (1-naphthyl) ethylamine (5.28 g,3.5 eq) and seed crystals (0.03 g), and the mixture was stirred at 25℃for 12 hours or more and then cooled to 0 ℃. Heptane (88 mL) was added dropwise over 1 hour, and after stirring for 2 hours, the precipitated crystals were filtered to obtain wet crystals (purity 92.9 PA%). To the wet crystals obtained, ethyl acetate (125 mL) was added, and after stirring at 35℃for 30 minutes, the mixture was cooled to 25℃and stirred for 12 hours. the slurry was cooled to 0deg.C over 1 hour, and heptane (75 mL) was added dropwise over 1 hour. After stirring at 0℃for 2 hours, the precipitated crystals were filtered and dried under reduced pressure to give 4-methoxyphenyl 2,3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (2-carboxybenzamide) - β -D-glucopyranosyl- (1.fwdarw.2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1.fwdarw.6) -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (2-carboxybenzamide) - β -D-glucopyranosyl- (1.fwdarw.4) -3 as white crystals, 6-di-0-benzyl-2-deoxy-2- (2-carboxybenzamide) -beta-D-glucopyranoside tris (R) - (+) -1- (1-naphthyl) ethylamine salt (compound represented by formula A-14) (18.93 g, yield 63.1%, purity 97.2 PA%). Powder X-ray crystal analysis of the obtained crystals is shown below.

< Measurement apparatus >)

Powder X-ray crystal analysis measuring device: rigaku.

< Measurement Condition >

Wavelength: cuka-

Goniometer: miniFlex 300/600.

Scanning speed: continuOUS.

Scan speed/count time: 10.00.

Width of steps: 0.02deg.

Scanning axis: 2 theta/theta.

Scanning range: 3.00-40.00 deg.

And (3) a filter: k-beta (. Times.1).

And (3) rotation: there are.

Powder X-ray crystallography determination of Compounds of formula A-14

Conversion of the Compound of formula A-14 to the Compound of formula A-13

In the case of 4-methoxyphenyl 2,3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl- (1→4) -3, 6-di-O-benzyl-2-deoxy-2- (2-carboxybenzamide) - β -D-glucopyranosyl- (1→2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1→6) -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1→4) -3, 6-di-O-benzyl-2-deoxy-2- (2-carboxybenzamide) - β -D-glucopyranosyl- (1→4) -3, 6-di-0-benzyl-2-deoxy-2- (2-carboxybenzamide) - β -D-glucopyranoside tris (R) - (+) -1- (1-naphtalenyl) ethylamine salt (the compound represented by formula A-14) (18.00 g, to 97.2 PA%) was added cyclopentyl methyl ether (90 mL), followed by three washes with 1M aqueous hydrochloric acid (180 mL) and 5% brine (90 mL). The resulting organic layer was concentrated to 18mL, cyclopentyl methyl ether (90 mL) was added, and concentrated to 18mL under reduced pressure. Tetrahydrofuran (90 mL) was added to the concentrate, and carbonyldiimidazole (6.86 g,8 eq) was added. After completion of the reaction (the reaction product obtained by conducting the ring closure reaction of phthalimide and the imidazole carbonylation of the hydroxyl group was confirmed) by stirring at 35℃for 1 hour, water (9 mL) and trifluoroacetic acid (12.1 g,20 equivalents) were added. Heating to 60 ℃, stirring for 20 hours, confirming that the deimidazole carbonylation is carried out, and cooling to room temperature. After ethyl acetate (90 mL) and water (90 mL) were added to separate the solutions, the organic layer was washed with 5% sodium bicarbonate water (90 mL) and water (90 mL) in this order, and the resulting organic layer was concentrated to 18mL under reduced pressure. Toluene (90 mL) was added and concentrated to 18mL, and methylene chloride (162 mL) was added. The methylene chloride solution was poured into silica gel (36 g, FUJI SILYSIA system ChromatorexSMB-20/45), and the elution was performed with a diisopropyl ether/methylene chloride mixed solvent (7/93, 720 mL) (fraction separation was performed every 90mL in the liquid-passing and elution operations, and the solution was recovered). The fractions were selected by performing purity measurement by HPLC, and the selected fractions were mixed and concentrated to 18mL. To the concentrated solution was added cyclopentyl methyl ether (90 mL), and after concentrating to 36mL, the solution was added dropwise to isopropyl alcohol (630 mL) cooled to 0 ℃ over 30 minutes with stirring. After stirring at 0℃for 2 hours, the slurry was filtered, washed with 0℃isopropyl alcohol (180 mL), and the resulting powder was dried under reduced pressure to give 4-methoxyphenyl 2,3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 6) -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1- > 4) -3 as a white powder, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranoside (compound of formula A-13) (14.1 g, yield 93%, purity 97.0 PA%).

[ Analysis conditions ]

Chromatographic column: xbridge Phenyl 3.5 μm, 4.6phi.times.150mm (Waters).

Wavelength: 220nm.

And (3) an oven: 40 ℃.

Eluent: (A) 10mM AcONH ₄ aqueous solution, (B) acetonitrile.

Gradient: 0 min (B) concentration 85%.

The concentration of 30 minutes (B) was 100%.

30.01 Min (B) concentration 85%.

35 Min (B) concentration 85%.

Flow rate: 1 mL/min.

And (3) sample injection: 5. Mu.L.

HRMS (ESI ⁺)[M+NH₄]⁺C₁₇₂H₁₆₈N₃O₃₅ theory: 2852.1718; experimental: 2852.1694).

¹H－NMR(500MHz,CDCl₃)δσ7.75(d,J＝8.0Hz,1H),7.69－7.40(10H),7.55－7.10(m,54H),7.06－6.97(5H),6.97－6.89(m,8H),6.88－6.84(m,1H),6.82－6.73(m,5H),6.72－6.67(m,2H),6.62－6.56(m,5H),5.42(d,J＝8.6Hz,1H),5.22(d,J＝8.0Hz,1H),5.03(d,J＝12.0Hz,1H),4.99(d,J＝8.6Hz,1H),4.92(d,J＝10.9Hz,1H),4.87－4.79(m,5H),4.75(d,J＝11.5Hz,1H),4.70(d,J＝12.0Hz,1H),4.67(d,J＝12.0Hz,1H),4.63(d,J＝6.3Hz,1H),4.61(d,J＝6.9Hz,1H),4.60－3.97(m,30H),3.88(d,J＝2.9Hz,1H),3.84(d,J＝12.0Hz,1H),3.82－3.67(m,5H),3.66－3.60(m,5H),3.54－3.33(m,14H),3.27－3.23(m,1H),3.20(d,J＝10.9Hz,1H),3.15－3.09(m,2H),2.83－2.78(m,1H),2.27－2.21(m,1H).

¹³C－NMR(125MHz,CDCl₃)δ168.4,168.2,167.4,167.3,155.2,150.8,139.1,139.0,138.9,138.8,138.7,138.51,138.49,138.44,138.38,138.37,138.35,138.26,138.0,137.7,133.8,133.6,122.49,133.41,133.36,131.8,131.4,128.6,128.52,128.49,128.39,128.34,128.30,128.2,128.14,128.10,128.06,128.0,127.85,127.82,127.7,127.69,127.65,127.56,127.53,127.48,127.38,127.33,127.26,127.1,127.0,126.9,126.7,123.5,123.2,123.09,123.05,122.98,118.5,114.2,102.9,102.6,97.8,97.4,97.0,96.9,82.5,80.5,79.9,78.5,77.8,77.4,76.9,76.7,75.87,75.80,75.2,75.0,74.79,74.77,74.62,74.59,74.48,74.41,74.2,74.0,73.9,73.6,73.4,73.0,72.9,72.7,72.62,72.56,72.0,69.8,68.24,68.20,68.1,67.8,67.2,56.6,55.6,55.5,53.4.

Example 29

Regarding the deacylation reaction of the compound represented by formula C-8 to the compound represented by formula C-9 in example 15 described above, comparative experiments of the deacylation reaction were conducted using the reaction conditions shown in the following table.

Items 1 to 3 are comparative examples, and items 4 and 5 are examples of the present invention.

TABLE 1

In the case of NaOMe, the ring-opening reaction proceeds completely due to the influence of moisture in the reagent (item 1).

Under acidic conditions, the cleavage proceeds with a slow reaction rate although the ring opening of the phthalimide group can be suppressed (item 2).

When t-BuOK is used, the target product is produced in about 85%, but about 8% of the ring opening is by-produced due to the influence of the moisture in the reagent or the solvent (item 3).

When CF ₃CO₂ Me was added, the reaction was terminated so as to suppress the opening of the ring almost completely. t-BuOK and LHMDS as bases gave equivalent results (items 4, 5).

Example 30

The debenzylation reaction of the compound of the formula A-12 containing 15 benzyl groups and 1 2-naphthylmethyl groups was carried out under the conventional conditions (CH ₂Cl₂－H₂ O) (item 1) and the method of the present invention (HFIP-H ₂ O) (items 2 and 3) as shown in the following table, and analyzed by HPLC to calculate the area peak ratio of the target compound (compound of the formula A-13) and debenzylated substrate as an excessive reactant.

The results are shown in the following table.

TABLE 2

Item 1: the selectivity to debenzylation substrate was moderate under the conditions of the conventional method.

Item 2: selectivity is improved by using HFIP.

Item 3: the best results were obtained by conducting the reaction at a lower temperature using HFIP.

Example 31

The area peak ratio of the target compound (compound represented by the following formula A-4') was calculated by analysis using HPLC, as shown in the following table, by performing the reaction of the compound represented by the formula A-4 containing 4 benzyl groups and 1-naphthylmethyl groups with respect to the 2-naphthylmethylation reaction under the conventional conditions (CH ₂Cl₂－H₂ O) (item 1), the process of the present invention (HFIP-H ₂ O) (item 2), the acidic conditions (item 3) and the hydrogenation conditions (item 4). The results are shown in the following table.

TABLE 3

Item 1: the reaction is carried out in moderate to high yields under the conditions of the prior art processes.

Item 2: the reaction is further improved by using HFIP.

Item 3: HCl/HFIP conditions reported in the paper (J. Org. Chem.2015, 80, 8796-8806) were applied, but the reaction was complicated, resulting in low yields.

Item 4: when hydrogenation conditions are used, a detachment of the benzyl group is also observed, resulting in low yields.

Example 32

The area peak ratio of the target (compound represented by the following formula A-10') was calculated by analysis using HPLC, as shown in the following table, by performing the reaction under the conventional condition (CH ₂Cl₂－H₂ O) (item 1) and the method of the present invention (HFIP-H ₂ O) (item 2) for the 2-naphthylmethylation removal of the compound represented by the formula A-10 containing 9 benzyl groups and 1 2-naphthylmethyl groups. The results are shown in the following table.

TABLE 4

Example 33

< Separation and purification of Compound represented by formula A-8 and Compound represented by formula A-9 >)

Examples of separation and purification of the compound represented by formula A-8 and the compound represented by formula A-9, which are sugar acceptors used in the synthesis of tetraose, are shown below. The compound represented by the formula A-8 as a sugar acceptor and the compound represented by the formula A-9 as a tetrasaccharide have very close polarities in normal phase silica gel column chromatography, and are difficult to separate, for example, at the same Rf value under hexane-ethyl acetate conditions as a typical column solvent system. By using the present application, monosaccharides and tetrasaccharides having extremely close polarities in silica gel can be easily separated.

The experimental procedure is as follows. Firstly, triethylamine is added into the solution after the reaction to stop the reaction, and then the molecular sieve is filtered, concentrated and acetonitrile is added. After octadecyl modified silica gel was added to the solution, water was added to adsorb the compound represented by the formula A-9 as a tetraose. When the filtrate was analyzed by HPLC, it was found that the compound represented by the formula A-9 was adsorbed as a tetraose and the compound represented by the formula A-8 was present in the filtrate as a monosaccharide. After the compound represented by the formula A-8 as a monosaccharide was washed by adding any acetonitrile-water, the compound represented by the formula A-9 as a tetrasaccharide was extracted with acetonitrile and toluene. The purification results of the compounds represented by the formulas A-8 are shown in tables 1 and 2 below.

TABLE 5

< Synthesis of Compound represented by formula D-3 >

The compound represented by the formula D-3 was synthesized according to the following synthesis pathway Z.

[ Synthetic Path Z ]

Example 34

4-Methoxyphenyl 4-O-acetyl-3, 6-di-O-benzyl-2-deoxy-2- (1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranoside (a compound represented by formula F-1)

(Small procedure Z-1)

To a solution of 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (a compound represented by formula A-8) (50.0 g,83.94 mmol) in ethyl acetate (200 mL) were added triethylamine (11.04 g,109.12 mmol), dimethylaminopyridine (0.31 g,2.52 mmol) and acetic anhydride (11.10 g,109.12 mmol), and the mixture was stirred at 20℃for 4 hours. After completion of the reaction, ethanol (500 mL) and water (150 mL) were added dropwise by HPLC. The slurry was stirred for 1 hour, the precipitated crystals were filtered, the filtered crystals were washed with a mixture of ethanol and water (150 mL/50 mL), and dried under reduced pressure at 40℃to give 4-methoxyphenyl 4-O-acetyl-3, 6-di-O-benzyl-2-deoxy-2- (1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranoside (compound represented by formula F-1) (49.0 g, yield 91%) as white crystals.

¹H－NMR(500MHz,CDCl₃)δ7.85－7.60(m,4H),7.24－7.34(m,5H),7.04－7.00(m,2H),6.96－6.87(m,3H),6.84(dt,J＝9.0,3.0Hz,2H),6.67(dt,J＝8.5,2.5Hz,2H),5.66(t,J＝4.0Hz,1H),5.22－5.18(m,1H),4.64(d,J＝12.0Hz,1H),4.55－4.48(m,4H),4.36(d,J＝12.0Hz,1H),3.90－3.84(m,1H),3.68(s,3H),3.68－3.64(m,2H),1.98(s,3H).

¹³C－NMR(125MHz,CDCl₃)δ169.6,155.3,150.6,137.8,137.5,133.9,128.2,128.0,127.7,127.7,127.5,127.4,123.3,118.4,114.3,97.4,76.8,73.9,73.7,73.5,72.2,69.4,55.4,55.3,20.8.

HRMS (ESI ⁺)[M+H]⁺C₃₇H₃₆NO₉ theory: 638.2385; experimental 638.2401).

Example 35

4-O-acetyl-3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -D-glucopyranoside (Compound represented by formula F-2)

(Small procedure Z-2)

To a solution of 4-methoxyphenyl 4-O-acetyl-3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (compound of formula F-1) (49.0 g,76.84 mmol) in dichloromethane (392 mL), hexafluoro-2-propanol (245 mL) and water (25 mL) at 25 ℃ was added [ bis (trifluoroacetoxy) iodo ] benzene (46.3 g,107.58 mmol) and stirred at the same temperature for 4 hours. After completion of the reaction was confirmed by HPLC, ethyl acetate (1225 mL) was added, and after ice-cooling, water (490 mL) in which sodium hydrogencarbonate (24.5 g) and sodium sulfite (24.5 g) were dissolved was poured and separated to obtain an organic layer. The resulting organic layer was again washed with water (490 mL) in which sodium bicarbonate (24.5 g) and sodium sulfite (24.5 g) were dissolved, and further washed with 20% brine (245 g). The obtained organic layer was concentrated under reduced pressure to 490mL (precipitation of crystals was confirmed during concentration), and heptane (735 mL) was added dropwise. The slurry obtained was cooled to 0℃to 5℃and stirred at the same temperature for 1 hour, and the precipitated crystals were filtered. The filtered crystals were washed with a mixture of ethyl acetate and heptane (39 mL/118 mL) at 0 to 5c and dried under reduced pressure at 40 c to give 4-O-acetyl-3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -D-glucopyranoside (compound represented by formula F-2) (37.5 g, yield 92%) as white crystals.

¹H－NMR(400MHz,CDCl₃)δ7.71－7.65(m,4H),7.34－7.26(m,5H),7.01－6.87(m,5H),5.36(dd,J＝8.0,8.0Hz,1H),5.13(dd,J＝8.4,10.0Hz,1H),4.59(d,J＝12.4Hz,1H),4.54(s,2H),4.50(dd,J＝8.4,10.4Hz,1H),4.33(d,J＝12.4Hz,1H),4.17(dd,J＝8.4,10.4Hz,1H),3.79(ddd,J＝8.4,5.2,4.8Hz,1H),3.61－3.53(m,2H),3.41(d,J＝8.0Hz,1H),1.93(s,3H).

¹³C－NMR(100MHz,CDCl₃)δ169.8,168.1,137.7,137.7,134.0,131.6,128.4,128.2,128.0,127.8,127.7,127.5,123.4,116.2,92.9,73.9,73.7,73.5,72.2,69.3,57.1,20.9.

HRMS (ESI ^－)[M－H]^－C₃₀H₂₈NO₈ theory: 530.1820; experimental 530.1841).

Example 36

4-O-acetyl-3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -O- [2, 2-trifluoro-N-phenylethanoimide ] -beta-D-glucopyranoside (a compound represented by formula D-3)

(Small procedure Z-3)

4-O-acetyl-3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -D-glucopyranoside (a compound represented by formula F-2) (20.0 g,37.63 mmol) is introduced into a 500mL eggplant-shaped flask, and dichloromethane (200 mL) and molecular sieve 4A powder (10 μm or less, 10.0 g) are added. N-methylimidazole (3.40 g,41.39 mmol) and 2, 2-trifluoro-N-phenyliminoacetyl chloride (8.20 g,39.51 mmol) were added sequentially under nitrogen at 0℃and stirred at the same temperature for 18 hours. After completion of the reaction was confirmed by HPLC, the reaction mixture was filtered through a filter and washed with methylene chloride (100 mL). The filtrate was filtered through a neutral silica gel pad (silica gel 60N, manufactured by Kanto chemical Co., ltd., particle size: 40 to 50 μm,60 g) filled with methylene chloride, and 100mL each time was taken. The silica gel pad was washed with dichloromethane (400 mL, 100mL each) and ethyl acetate/dichloromethane (1:4, 400mL, 100mL each) and the selected fractions were concentrated to a liquid volume of 40mL. Toluene (200 mL) was added thereto, and the mixture was concentrated again to a liquid volume of 40mL, and toluene (200 mL) was further added thereto, and the concentrated mixture was concentrated to a liquid volume of 40mL, whereby a crude product of 4-O-acetyl-3, 6-di-O-benzyl-2-deoxy-2- (1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) -O- [2, 2-trifluoro-N-phenylethanimido ] -beta-D-glucopyranoside (a compound represented by formula D-3) was obtained as a toluene solution. The product was used directly in the next step.

< Synthesis of Compound represented by formula D-7 >

The compound represented by the formula D-7 was synthesized according to the following synthesis pathway V.

[ Synthesis pathway V ]

First, a compound represented by the formula G-1 was synthesized according to the following synthesis pathway W.

[ Synthesis path W ]

Example 37

Allyl alpha-D-galactopyranoside (compound represented by G-0)

After D-galactopyranose (20.00 g,111.01 mmol) was added to a 500mL four-necked flask, allyl alcohol (200.0 mL) was added. p-TsOH H ₂ O (2.11 g,11.10 mmol) was added at room temperature under nitrogen atmosphere, warmed to 70℃and stirred for 24 hours. The reaction mixture was cooled to 40℃and triethylamine (1.69 g,16.65 mmol) was added thereto and stirred for 5 minutes, followed by concentrating the reaction mixture under reduced pressure to a liquid volume of 100mL. To the concentrate was added nBuOH (200 mL) dropwise over 30 minutes, and the mixture was stirred at room temperature for 1 hour. Thereafter, the reaction solution was concentrated under reduced pressure to 80mL in a liquid volume, and stirred at room temperature overnight. The suspension was filtered, and the crystals were washed with nBuOH (40 mL) at 0 ℃ and dried under reduced pressure at 40 ℃ to give allyl α -D-galactopyranoside (compound represented by G-0) as white crystals (8.41G, 34.4% yield).

¹H－NMR(500MHz,METHANOL－D4)δ5.97(qd,J＝11.2,5.7Hz,1H),5.33(dd,J＝17.2,1.7Hz,1H),5.17(dd,J＝10.6,1.4Hz,1H),4.22(dd,J＝13.2,5.2Hz,1H),4.04(dd,J＝13.0,6.0Hz,1H),3.88(d,J＝1.7Hz,1H),3.82－3.66(m,5H).

¹³C－NMR(125MHz，METHANOL－D4)δ62.74，69.36，70.21，71.08，71.51，72.49，99.46，117.47，135.69。

MS(ESI)m/z：221(M+H)⁺,219(M－H)^－。

Example 38

Prop-2-en-1-yl 4, 6-O-benzylidene-alpha-D-galactopyranoside (a compound represented by formula G-1)

After acetonitrile (5.0 mL), benzaldehyde dimethyl acetal (5.18 g,34.1 mmol) and p-TsOH.H ₂ O (431.9 mg,2.27 mmol) were added to a 100mL eggplant-shaped flask under nitrogen, allyl alpha-D-galactopyranoside (5.00 g,22.7 mmol) was added in small portions. The solution was warmed to 40 ℃ and stirred for 30 minutes. Then, triethylamine (344.6 mg,3.41 mmol) was added thereto and stirred for 5 minutes, followed by dropping isopropyl alcohol (75 mL) at 40 ℃. The reaction solution was concentrated under reduced pressure to 25mL in an amount, and stirred at 0℃overnight. Subsequently, the suspension was filtered, and the obtained crystals were washed with isopropyl alcohol (5 mL) cooled to 0 ℃ and dried under reduced pressure at 40 ℃ to obtain prop-2-en-1-yl 4, 6-O-benzylidene- α -D-galactopyranoside (the compound represented by formula G-1) (5.35G, yield 76.3%) as white crystals.

¹H－NMR(500MHz,METHANOL－D4)δ7.53－7.52(dd,J＝7.5,2.0Hz,2H),7.34(m,3H),5.98(qd,J＝11.1,5.4Hz,1H),5.59(s,1H),5.35(d,J＝18.9Hz,1H),5.19(d,J＝10.3Hz,1H),4.95(d,J＝4.0Hz,1H),4.26(d,J＝3.4Hz,1H),4.22(dd,J＝13.2,5.2Hz,1H),4.13(s,2H),4.08(q,J＝6.5Hz,1H),3.92(ddd,J＝24.1,10.3,3.4Hz,2H),3.74(s,1H).

¹³C－NMR(125MHz,METHANOL－D4)δ64.60,69.70,70.03,70.08,70.34,78.07,100.22,102.28,117.58,127.54,129.02,129.86,135.59,139.77.

MS(ESI)m/z：309(M+H)⁺,307(M－H)^－。

Example 39

Prop-2-en-1-yl 2, 3-di-O-benzoyl-4, 6-O-benzylidene-alpha-D-galactopyranoside (a compound represented by formula G-2)

(Small procedure V-1)

Benzoyl chloride (47.87G, 340.54 mmol) was added dropwise to a solution of prop-2-en-1-yl 4, 6-O-benzylidene-alpha-D-galactopyranoside (compound of formula G-1) (30.0G, 97.30 mmol) in pyridine (150 mL) at 40℃or below, and stirred at the same temperature for 2 hours. After confirming the completion of the reaction by HPLC, the reaction mixture was cooled to 20℃to 30℃and ethanol (450 mL) was injected, followed by dropwise addition of water (300 mL) over a period of 30 minutes. After stirring the slurry at 20℃to 30℃for 1 hour, the precipitated crystals were filtered, and the filtered crystals were washed with a mixture of ethanol and water (75 mL/75 mL), and dried under reduced pressure at 40℃to give prop-2-en-1-yl 2, 3-di-O-benzoyl-4, 6-o-benzylidene-alpha-D-galactopyranoside (compound represented by formula G-2) (47.9G, yield 95%) as white crystals.

¹H－NMR(500MHz,CDCl₃)δ8.03－7.98(m,4H),7.55－7.46(m,4H),7.40－7.30(m,7H),5.90－5.78(m,2H),5.82(s,1H),5.57(s,1H),5.42(d,J＝1.7Hz,1H),5.31(dd,J＝17.2,1.7Hz,1H),5.15(dd,J＝1.7,10.5Hz,1H),4.66(s,2H),4.33(d,J＝12.5Hz,1H),4.26(dd,J＝12.5,4.5Hz,1H),4.12(dd,J＝12.5,1.0Hz,1H),4.08(dd,J＝6.5,13.5Hz,1H),3.95(s,1H).

¹³C－NMR(125MHz,CDCl₃)δ166.1,165.8,137.5,133.4,133.2,133.1,129.8,129.7,129.5,129.4,128.8,128.3,128.1,126.1,117.5,100.6,96.2,74.2,69.3,69.1,68.7,68.6,62.4.

HRMS (ESI ⁺)[M+H]⁺C₃₀H₂₉O₈ theory: 517.1857; experimental 517.1880).

Example 40

Prop-2-en-1-yl 2, 3-di-O-benzoyl-alpha-D-galactopyranoside (Compound represented by formula G-3)

(Small procedure V-2)

A solution of prop-2-en-1-yl 2, 3-di-O-benzoyl-4, 6-O-benzylidene-alpha-D-galactopyranoside (compound of formula G-2) (47.1G, 91.18 mmol) in acetonitrile (377 mL) was heated to 45℃and water (24 mL) and concentrated hydrochloric acid (9.2G, 91.18 mmol) were added and stirred at the same temperature for 30 minutes. Water (353 mL) was added dropwise over 3 hours at 45℃to 50℃and then stirred for a further 30 minutes. After completion of the reaction was confirmed by HPLC, sodium acetate (11.22 g,136.78 mmol) was added, ethyl acetate (942 mL) and water (471 mL) were poured, and the mixture was cooled to 25℃or lower, followed by separation to obtain an organic layer. The resulting organic layer was washed twice with water (471 mL) and then further washed with 20% brine (236 mL). The organic layer was concentrated to 141mL under reduced pressure, toluene (707 mL) was added, and concentrated again to 141mL under reduced pressure. Toluene (236 mL) was added to the resulting concentrate, and the mixture was concentrated under reduced pressure to 141mL. After cooling the concentrated solution to 0 to 5 ℃, a slurry of toluene (330 mL) containing neutral silica gel (silica gel 60N, manufactured by kanto chemical Co., ltd., particle diameter: 40 to 50 μm,141 g) cooled to 0 to 5 ℃ was injected, and the mixture was stirred at the same temperature for 15 minutes to adsorb the product on the silica gel, followed by filtration. After washing the silica gel solid phase containing the product with toluene (942 mL) at 0℃to 5℃and discarding the filtrate at the time of toluene washing, the target was desorbed from the silica gel with cyclopentyl methyl ether (707 mL) to obtain a cyclopentyl methyl ether solution (quantitative 36.2G, quantitative yield 93%) of prop-2-en-1-yl-2, 3-di-O-benzoyl-alpha-D-galactopyranoside (compound represented by the formula G-3). The solution was used in the next step.

Example 41

5-Acetamido-3, 5-dideoxy-D-glycero-D-galacto-non-2-pyronic acid methyl ester monohydrate (Compound represented by formula G-5)

(Small procedure V-3)

To a solution of 5-acetamido-3, 5-dideoxy-D-glycero-D-galacto-non-2-pyronic acid (ulopyranosonic acid) (compound of formula G-4) (40.1G, 129.66 mmol) and methyl orthoformate (15.60 mL,142.59 mmol) in methanol (321 mL) was added sulfuric acid (1.0G, 10.20 mmol), and the mixture was heated to 40℃and stirred for 3 hours. After confirming the completion of the reaction by HPLC, the reaction mixture was cooled to 25℃and dimethylacetamide (40 mL) was added thereto, followed by concentration under reduced pressure to 160mL. The temperature of the resulting concentrate was adjusted to 15 ℃, water (20 mL) and ethyl acetate (722 mL) were injected, and after stirring at 25 ℃ for 1 hour, the slurry was cooled to 0 ℃ to 5 ℃ and stirred at the same temperature for 2 hours. The precipitated crystals were filtered, washed with ethyl acetate (80 mL) at 0℃to 5℃and dried under reduced pressure at 40℃to give methyl 5-acetamido-3, 5-dideoxy-D-glycero-D-galacto-non-2-pyronate monohydrate (compound represented by the formula G-5) (41.1G, yield 93%) as white crystals.

¹H－NMR(500MHz,CD₃OD)δ4.07－3.98(m,2H),3.85－3.77(m,2H),3.78(s,3H),3.72－3.68(m,1H),3.62(dd,J＝10.9,5.7Hz,1H),3.48(dd,J＝9.2,1.1Hz,1H),2.22(dd,J＝12.9,4.9Hz,1H),2.02(s,3H),1.89(dd,J＝12.6,11.5Hz,1H).

¹³C－NMR(125MHz,CD₃OD)δ175.2,175.1,171.8,96.6,72.1,72.0,71.6,70.1,67.9,64.8,54.4,54.3,53.2,40.7,22.7,22.7.

HRMS (ESI ⁺)[M+H]⁺C₁₂H₂₂NO₉ theory: 324.1289; experimental 324.1288).

Example 42

5-Acetamido-4, 7,8, 9-tetra-O-acetyl-3, 5-dideoxy-D-glycero-D-galacto-non-2-pyronic acid methyl ester (Compound represented by formula G-6)

(Small procedure V-4)

Methyl 5-acetamido-3, 5-dideoxy-D-glycero-D-galacto-non-2-pyronate monohydrate (compound of formula G-5) (40.3G, 118.07 mmol) was adjusted to a slurry temperature of 25℃and acetic anhydride (60.27G, 590.36 mmol) and p-toluenesulfonic acid monohydrate (1.12G, 5.89 mmol) were added and stirred at 25℃for 24 hours. After that, the reaction solution was cooled to 15℃and acetic anhydride (12.05 g,118.03 mmol) was added thereto, followed by stirring at the same temperature for 47 hours. After completion of the reaction by HPLC, methanol (40 mL) was added thereto, the temperature was adjusted to 25℃and the mixture was stirred at the same temperature for 2 hours. Next, sodium acetate (0.97 g,11.82 mmol) was added thereto, and further stirred at the same temperature for 1 hour. The reaction mixture was concentrated to 120mL under reduced pressure, cooled to 0℃to 5℃and then poured into ethyl acetate (403 mL) and water (161 mL), and triethylamine was added thereto under stirring at 0℃to 5℃to adjust the pH to 7.0. The organic layer obtained by separation was washed twice with 10% brine (121 mL), and concentrated to 200mL under reduced pressure. To the concentrate was added ethyl acetate (605 mL), and the mixture was concentrated again under reduced pressure to 200mL. To the concentrate was added ethyl acetate (40 mL), and after seeding with seed crystals and stirring at 25℃for 4 hours, heptane (302 mL) was added dropwise over 30 minutes. After stirring the slurry at 25℃for 2 hours, the precipitated crystals were filtered, and the filtered crystals were washed with a mixture of ethyl acetate and heptane (67 mL/135 mL), and dried under reduced pressure at 35℃to give methyl 5-acetamido-4, 7,8, 9-tetra-O-acetyl-3, 5-dideoxy-D-glycero-D-galacto-non-2-pyronate (compound represented by formula G-6) (44.1G, yield 76%) as white crystals.

¹H－NMR(500MHz,CDCl₃)δ6.28(d,J＝10.3Hz,1H),5.41(dd,J＝4.6,2.3Hz,1H),5.28－5.23(m,1H),5.21－5.14(m,1H),5.09(s,1H),4.62(dd,J＝12.3,2.6Hz,1H),4.28(dd,J＝10.3,2.3Hz,1H),4.21－4.11(m,1H),4.04(dd,J＝12.3,8.3Hz,1H),3.85(s,3H),2.24－2.20(m,2H),2.16(s,3H),2.12(s,3H),2.03(s,3H),2.01(s,3H),1.91(s,3H).

¹³C－NMR(125MHz,CDCl₃)δ171.5,171.1,170.8,170.3,170.2,168.9,94.9,72.1,71.4,69.1,68.3,62.5,53.2,49.1,36.1,23.0,21.0,20.8,20.7,20.7.

HRMS (ESI ⁺)[M+H]⁺C₂₀H₃₀NO₁₃ theory: 492.1712; experimental 492.1712).

Example 43

5-Acetamido-4, 7,8, 9-tetra-O-acetyl-3, 5-dideoxy-2-O- (2, 2-trifluoro-N-phenylethanimide) -D-glycero-beta-D-galacto-non-2-pyronate methyl ester (Compound represented by formula G-7)

(Small procedure V-5)

Methyl 5-acetamido-4, 7,8, 9-tetra-O-acetyl-3, 5-dideoxy-D-glycero-D-galacto-non-2-pyronate (compound represented by formula G-6) (44.0G, 89.53 mmol) and molecular sieve 4A powder (powder particle size 10 μm or less) (22G) were stirred at the same temperature for 30 minutes after adjusting the methylene chloride (352 mL) slurry temperature to 20℃and adding 2, 2-trifluoro-N-phenyliminoacetyl chloride (26.02G, 125.35 mmol). Next, N-methylimidazole (11.03 g,134.33 mmol) was added dropwise thereto, and the mixture was stirred at 20℃for 7.5 hours. After completion of the reaction was confirmed by HPLC, the reaction solution was filtered and washed with methylene chloride (88 mL) to obtain a filtrate. The filtrate was cooled to 0℃and cold water (440 mL) was added thereto, and triethylamine was added thereto under stirring at 0℃to 5℃to adjust the pH to 7.5. After stirring at 0℃to 5℃for 30 minutes, the resulting organic layer was separated, washed twice with cold water (440 mL), washed with cooled 20% brine (220 mL), and concentrated to 88mL under reduced pressure. To the concentrate was added ethyl acetate (440 mL), and the mixture was concentrated again under reduced pressure to 88mL. To the concentrate was added tert-butyl methyl ether (308 mL), and the mixture was seeded and stirred at 20℃for 4 hours. To the resulting slurry was added heptane (264 mL) dropwise over 1 hour, stirred at the same temperature for 2 hours, and the precipitated crystals were filtered, washed with a mixture of t-butyl methyl ether and heptane (132 mL/88 mL), and dried under reduced pressure at 35℃to give 5-acetamido-4, 7,8, 9-tetra-O-acetyl-3, 5-dideoxy-2-O- (2, 2-trifluoro-N-phenylethanimido) -D-glycero- β -D-galacto-non-2-pyronate methyl ester (compound represented by formula G-7) (39.5G, yield 67%) as white crystals.

¹H－NMR(500MHz,CDCl₃)δ7.30－7.24(m,2H),7.09(t,J＝7.4Hz,1H),6.72(d,J＝8.0Hz,2H),5.76(d,J＝9.7Hz,1H),5.48－5.45(m,1H),5.30(td,J＝10.9,4.8Hz,1H),5.15－5.10(m,1H),4.60(dd,J＝12.6,2.3Hz,1H),4.30(q,J＝10.3Hz,1H),4.23(dd,J＝10.3,2.3Hz,1H),4.11(dd,J＝12.3Hz,7.7Hz,1H),3.81(s,3H),2.79(dd,J＝13.5,4.9Hz,1H),2.21－2.15(m,1H),2.16(s,3H),2.10(s,3H),2.07(s,3H),1.90(s,3H),1.75(s,3H).

¹³C－NMR(125MHz,CDCl₃)δ171.0,170.7,170.4,170.2,170.1,165.3,142.6,141.0(q,²J_C－F＝36.0Hz),128.8,124.6,119.0,115.1(q,¹J_C－F＝284.4Hz),99.7,73.6,71.9,68.3,68.0,62.4,53.1,48.6,35.6,23.0,20.8,20.8,20.7,20.3.

HRMS (ESI ⁺)[M+NH₄]⁺C₂₈H₃₇F₃N₃O₁₃ theory: 680.2273; experimental 680.2314).

Example 44

4,7,8, 9-Tetra-O-acetyl-5- [ acetyl (t-butoxycarbonyl) amino ] -3, 5-dideoxy-2-O- (2, 2-trifluoro-N-phenylethanimide) -D-glycero-beta-D-galacto-non-2-pyronic acid methyl ester (Compound represented by formula G-8)

(Small procedure V-6)

To a solution of 5-acetamido-4, 7,8, 9-tetra-O-acetyl-3, 5-dideoxy-2-O- (2, 2-trifluoro-N-phenylethanimido) -D-glycero-beta-D-galacto-non-2-pyronate (compound of formula G-7) (39.0G, 58.86 mmol) in tetrahydrofuran (390 mL) were added di-tert-butyl dicarbonate (27.05G, 123.94 mmol) and dimethylaminopyridine (1.80G, 14.73 mmol), and the mixture was heated to reflux temperature. After stirring at reflux for 30 minutes, the completion of the reaction was confirmed by HPLC, and the reaction solution was concentrated to 117mL under reduced pressure. Toluene (195 mL) was added to the concentrate, and the mixture was concentrated again under reduced pressure to 117mL. The concentrated solution was filtered using a funnel (silica gel 60N, manufactured by Kanto chemical Co., ltd., particle diameter: 40 to 50 μm,117g, toluene wet-packed) filled with silica gel, and washed with toluene/ethyl acetate mixture (8/2) (975 mL) to obtain a filtrate. The resulting filtrate was concentrated under reduced pressure (to a weight of 59 g), and cyclopentyl methyl ether (23 mL) was added. The solution temperature was adjusted to 20℃and heptane (156 mL) was added dropwise over 15 minutes, followed by stirring at the same temperature for 1 hour. After confirming the precipitation of crystals, heptane (312 mL) was added dropwise over 1 hour, the precipitated crystals were filtered, the filtered crystals were washed with heptane (78 mL), and the filtered crystals were dried under reduced pressure at 35℃to obtain methyl 4,7,8, 9-tetra-O-acetyl-5- [ acetyl (t-butoxycarbonyl) amino ] -3, 5-dideoxy-2-O- (2, 2-trifluoro-N-phenylethanimido) -D-glycero- β -D-galacto-non-2-pyronate (compound represented by formula G-8) (37.0G, yield 82%) as white crystals.

¹H－NMR(500MHz,CDCl₃)

Note) was detected as a ca.1/4 isomer mixture. Major isomer ：δ7.27(t,J＝8.9Hz,2H),7.09(t,J＝7.2Hz,1H),6.73(d,J＝8.0Hz,2H),5.75－5.65(m,1H),5.31(d,J＝4.6Hz,1H),5.18－5.14(m,1H),5.15(d,J＝6.0Hz,2H),4.54(dd,J＝12.0,2.0Hz,1H),4.08(dd,J＝12.6,6.9Hz,1H),3.84(s,3H),2.90(dd,J＝13.7,5.2Hz,1H),2.39(s,3H),2.25(dd,J＝13.7,11.2Hz,1H),2.09(s,3H),2.07(s,3H),1.99(s,3H),1.77(s,3H),1.62(s,9H). minor isomer ：δ6.76(d,J＝8.0Hz,2H),5.85－5.80(m,1H),5.29－5.25(m,1H),5.22－5.19(m,1H),4.44(d,J＝11.0Hz,1H),4.15－4.11(m,1H),3.03(dd,J＝14.0,5.0Hz,1H),2.41(s,3H),2.12(s,3H),2.00(s,3H),1.88(s,3H),1.74(s,3H),1.54(s,9H).

¹³C－NMR(125MHz,CDCl₃ ) Mixture ：δ173.7,170.4,170.2,170.0,169.9,165.4,151.7,142.8,128.7,124.5,119.1,100.6,85.2,72.8,71.3,67.7,65.9,62.0,53.1,52.0,36.7,27.9,27.7,26.6,20.8,20.7,20.6,20.3.

HRMS (ESI ⁺)[M+NH₄]⁺C₃₃H₄₅F₃N₃O₁₅ theory: 780.2797; experimental 780.2801).

Example 45

Prop-2-en-1-yl 2, 3-di-O-benzoyl-6-O- {4,7,8, 9-tetra-O-acetyl-5- [ acetyl (t-butoxycarbonyl) amino ] -3, 5-dideoxy-1-methyl-D-glycero-alpha-D-galacto-non-2-pyron-yl } -alpha-D-galactopyranoside (a compound represented by formula G-9)

(Small procedure V-7)

After concentrating a solution of prop-2-en-1-yl 2, 3-di-O-benzoyl-alpha-D-galactopyranoside (the compound represented by the formula G-3) in cyclopentyl methyl ether (quantitative 31.46G,73.43 mmol) under reduced pressure to 105mL, it was added to a solution of methyl cyclopentyl methyl ether (175 mL) of 4,7,8, 9-tetra-O-acetyl-5- [ acetyl (tert-butoxycarbonyl) amino ] -3, 5-dideoxy-2-O- (2, 2-trifluoro-N-phenylethanimido) -D-glycero-beta-D-galacto-non-2-pyronate (the compound represented by the formula G-8) (35.0G, 45.89 mmol). Then, cyclopentyl methyl ether was added to the obtained mixed solution, and the total amount was adjusted to 350mL (cyclopentyl methyl ether mixed solution of the compound represented by formula G-3 and the compound represented by formula G-8). In a separate vessel were added cyclopentyl methyl ether (525 mL) and molecular sieve 4A powder (powder particle diameter: 10 μm or less) (17.5 g), and after cooling to-60 ℃, trimethyl silyl triflate (4.2 mL,23.24 mmol) was added. To this solution, a mixed solution of a compound represented by formula G-3 and cyclopentylmethyl ether of a compound represented by formula G-8 was added dropwise over 4.5 hours at-60℃under vigorous stirring, and stirred at the same temperature for 2 hours. After completion of the reaction was confirmed by HPLC, triethylamine (4.5 mL,32.12 mmol) was added, and the reaction mixture was warmed to 0 ℃. After that, diatomaceous earth 545 (35.00 g) was added thereto, and the reaction solution was filtered and washed with cyclopentyl methyl ether (175 mL). Water (350 mL) was added to the filtrate to separate the solution. Subsequently, 0.5N hydrochloric acid water (350 mL) was added to the organic layer, and the mixture was stirred at 20℃for 2 hours. After confirming the decomposition of the by-product by HPLC, the organic layer was separated. The organic layer was washed with water (350 mL) and 20% brine (175 mL), and concentrated to 70mL under reduced pressure. Toluene (700 mL) was added to the concentrated solution, and the mixture was concentrated under reduced pressure to 70mL. Toluene (700 mL) and neutral silica gel (silica gel 60N, manufactured by Kanto chemical Co., ltd., particle size: 40-50 μm,158 g) were again added to the concentrated solution, and stirred at 20℃for 30 minutes. The product was adsorbed on silica gel, and then, the solid phase of the silica gel containing the product was washed with toluene (1575 mL) (filtrate at the time of toluene washing was discarded), and then, the target substance was desorbed from the silica gel with ethyl acetate (875 mL). The ethyl acetate solution obtained was concentrated under reduced pressure to 70mL, toluene (175 mL) was added and concentrated again under reduced pressure to 70mL, to give a toluene solution of prop-2-en-1-yl-2, 3-di-O-benzoyl-6-O- {4,7,8, 9-tetra-O-acetyl-5- [ acetyl (t-butoxycarbonyl) amino ] -3, 5-dideoxy-1-methyl-D-glycero-alpha-D-galacto-non-2-pyrone } -alpha-D-galactopyranoside (a compound represented by formula G-9). The solution was used in the next step.

Example 46

Prop-2-en-1-yl 6-O- { 5-acetamido-4, 7,8, 9-tetra-O-acetyl-3, 5-dideoxy-1-methyl-D-glycero-alpha-D-galacto-non-2-pyrone glycosyl } -2, 3-di-O-benzoyl-alpha-D-galactopyranoside (a compound represented by formula G-10)

(Small procedure V-8)

To a toluene solution of prop-2-en-1-yl 2, 3-di-O-benzoyl-6-O- {4,7,8, 9-tetra-O-acetyl-5- [ acetyl (t-butoxycarbonyl) amino ] -3, 5-dideoxy-1-methyl-D-glycero- α -D-galacto-non-2-pyron osyl } - α -D-galactopyranoside (a compound represented by formula G-9) obtained in example 45 were added dichloromethane (525 mL) and copper (II) triflate (8.30G, 22.95 mmol), and the mixture was warmed to 40 ℃ and stirred at the same temperature for 2 hours. After confirming the completion of the reaction by HPLC, the reaction mixture was cooled to 25℃and concentrated to 70mL under reduced pressure. To the concentrate was added ethyl acetate (525 mL), and after washing three times with 5% brine (350 mL), the organic layer was washed four times with 20% methanol water (525 mL) and heptane (263 mL) was added. After confirming by HPLC that the impurity derived from the beta-exosome of compound 8, which is a byproduct of the glycosylation reaction, was removed to an aqueous layer, the organic layer was concentrated to 70mL under reduced pressure. To the concentrated solution was added isopropenyl acetate (525 mL), and the mixture was concentrated under reduced pressure to 350mL to obtain an isopropenyl acetate solution of prop-2-en-1-yl 6-O- { 5-acetamido-4, 7,8, 9-tetra-O-acetyl-3, 5-dideoxy-1-methyl-D-glycero-alpha-D-galacto-non-2-pyron-osyl } -2, 3-di-O-benzoyl-alpha-D-galactopyranoside (a compound represented by formula G-10). The solution was used in the next step.

Example 47

Prop-2-en-1-yl 4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero-alpha-D-galacto-non-2-pyrone glycosyl ] -alpha-D-galactopyranoside (a compound represented by formula G-11)

(Small procedure V-9)

To the isopropyl acetate solution of prop-2-en-1-yl 6-O- { 5-acetamido-4, 7,8, 9-tetra-O-acetyl-3, 5-dideoxy-1-methyl-D-glycero- α -D-galacto-non-2-pyrone glycosyl } -2, 3-di-O-benzoyl- α -D-galactopyranoside (the compound represented by formula G-10) obtained in example 46 was added p-toluenesulfonic acid monohydrate (0.88G, 4.62 mmol), and the mixture was warmed to reflux temperature (about 90 ℃ c.) and stirred at the same temperature for 3 hours. After confirming the completion of the reaction by HPLC, the reaction mixture was cooled to 25℃and triethylamine (0.95 mL,6.85 mmol) was added thereto, followed by concentration under reduced pressure to 70mL. Toluene (350 mL) was added to the concentrate, and the mixture was concentrated again under reduced pressure to 70mL. Toluene (630 mL) was added to the concentrated solution, neutral silica gel (silica gel 60N, manufactured by Kanto chemical Co., ltd., particle diameter: 40 to 50 μm,123 g) was added, and the mixture was stirred at the same temperature for 30 minutes to adsorb the product on the silica gel, followed by filtration. After washing the silica gel solid phase containing the product with toluene (1925 mL) and toluene/ethyl acetate mixture (97/3, 1400 mL) (filtrate discarded during washing), the objective was desorbed from the silica gel solid phase containing the product with ethyl acetate (1050 mL), and the resulting ethyl acetate solution was concentrated under reduced pressure, whereby prop-2-en-1-yl 4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero- α -D-galacto-2-galacto-non-pyron-yl ] - α -D-galacto-pyranoside (compound represented by formula G-11) (30.1G, yield 67% (based on the compound represented by formula G-8)) was obtained as a white foam solid (comprising 0.42 equivalent of toluene (about 4 wt%)).

¹H－NMR(500MHz,CDCl₃)δ7.99(d,J＝8.4Hz,2H),7.88(dd,J＝8.4,1.4Hz,2H),7.53－7.47(m,2H),7.37(dt,J＝14.7,6.9Hz,4H),5.90－5.82(m,1H),5.82(dd,J＝10.9,3.4Hz,1H),5.73(d,J＝2.9Hz,1H),5.58(dd,J＝10.9,4.0Hz,1H),5.51(td,J＝10.6,5.0Hz,1H),5.35－5.30(m,3H),5.17－5.15(m,2H),4.94(dd,J＝10.3,1.7Hz,1H),4.38－4.27(m,3H),4.20－4.13(m,2H),4.08(dd,J＝13.2,5.7Hz,1H),3.94(dd,J＝10.3,6.3Hz,1H),3.82(s,3H),3.50(dd,J＝9.7,7.4Hz,1H),2.73(dd,J＝13.2,5.2Hz,1H),2.37(s,3H),2.31(s,3H),2.19(s,3H),2.15(s,3H),2.14(s,3H),2.03(s,3H),1.97(s,3H),1.86(dd,J＝13.2,10.9Hz,1H).

¹³C－NMR(125MHz,CDCl₃)δ174.5,173.6,170.5,170.1,169.9,169.8,169.6,167.3,166.0,165.5,133.5,133.3,133.1,129.8,129.5,129.4,128.4,128.3,117.5,98.6,95.5,77.2,69.8,68.9,68.6,68.6,68.3,68.3,67.5,67.0,66.7,62.4,61.8,57.0,52.9,38.7,27.9,25.9,21.0,20.9,20.7,20.7,20.6.

HRMS (ESI ⁺)[M+H]⁺C₄₇H₅₆NO₂₂ theory: 986.3288; experimental 986.3277).

For the obtained compounds, it was confirmed that the spectra are consistent with the following documents: j.org.chem.2016, 81, 10600-10616.

Example 48

4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero-alpha-D-galacto-no-2-pyron-osyl ] -D-galactopyranose (Compound represented by formula G-12)

(Small procedure V-10)

Five times of decompression and nitrogen substitution were repeated for a methanol solution (290 mL) of prop-2-en-1-yl 4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero-alpha-D-galacto-non-2-pyranosyl ] -alpha-D-galactopyranoside (compound represented by formula G-11) (29.00G, 29.41 mmol), 1, 3-dimethylbarbituric acid (9.19G, 58.86 mmol) and triphenylphosphine (2.31G, 8.81 mmol), and after degassing operation, palladium (II) acetate (0.66G, 2.94 mmol) was added and stirred at 40℃for 12 hours. After completion of the reaction was confirmed by HPLC, toluene (580 mL) and water (1015 mL) were added and the mixture was separated to obtain an organic layer. The organic layer was washed four times with 20% methanol water (580 mL), and after removing 1, 3-dimethylbarbituric acid to the aqueous layer, the solution was concentrated under reduced pressure to 58mL, toluene (435 mL) was added, and the solution was concentrated again under reduced pressure to 58mL. Toluene (383 mL), chloroform (197 mL) and neutral silica gel (silica gel 60N, manufactured by Kanto chemical Co., ltd., particle size: 40 to 50 μm,145 g) were added to the concentrated solution, and the mixture was stirred for 30 minutes to adsorb the product on the silica gel, followed by filtration. After washing the silica gel solid phase containing the product with toluene/chloroform mixture (2/1, 4350 mL) (filtrate waste during washing), the target substance was desorbed from the silica gel solid phase containing the product with ethyl acetate (870 mL). SH silica gel (29.00 g) was added to the ethyl acetate solution, and after stirring for 30 minutes, filtration was performed, and washing with ethyl acetate (145 mL) was performed to obtain an ethyl acetate solution containing the target substance. The resulting solution was concentrated to 58mL under reduced pressure, toluene (145 mL) was added, and concentrated again to 58mL under reduced pressure. The concentrated solution was purified by column chromatography on silica gel (silica gel 60N, manufactured by Kanto chemical Co., ltd., particle size: 40 to 50 μm,290g, mobile phase hexane/ethyl acetate 50/50 to 30/70), and the selected fraction was concentrated to 29mL under reduced pressure. To the concentrated solution were added ethyl acetate (290 mL) and activated carbon (14.5 g) and, after stirring for 30 minutes, filtration was performed, and washing with ethyl acetate (87 mL) was performed to obtain a purified ethyl acetate solution containing the target substance. The resulting solution was concentrated under reduced pressure to obtain 4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero- α -D-galacto-non-2-pyron-yl ] -D-galactopyranose (compound represented by formula G-12) (18.70G, yield 67%) as a white foam solid.

¹H－NMR(500MHz,CDCl₃ ) The major isomer ：δ7.98－8.03(m,2H),7.89(t,2H,J＝8.9Hz),7.52－7.48(m,2H),7.39－7.34(m,4H),5.92(dd,1H,J＝10.3,2.9Hz),5.82(d,1H,J＝3.4Hz),5.68(t,1H,J＝2.3Hz),5.59－5.48(m,2H),5.37(td,1H,J＝7.5,2.5Hz),5.17(d,1H,J＝7.5Hz),5.02(d,1H,J＝10.5Hz),4.74－4.71(m,1H),4.44－4.38(m,1H),4.16－4.08(m,2H),3.85(s,3H),3.80(m,1H),3.60－3.54(m,1H),2.76(dd,1H,J＝13.0,6.0Hz),2.38(s,3H),2.33(s,3H),2.31(s,3H),2.15(s,3H),2.12(s,3H),2.04(s,3H),1.98(s,3H),1.93－1.87(m,1H).

¹³C－NMR(125MHz,CDCl₃ ) Alpha/beta mixture ：174.5,173.8,173.6,171.7,171.0,170.5,170.3,170.3,169.9,169.8,169.8,169.6,167.6,167.3,166.3,166.0,165.6,165.4,133.3,133.2,133.2,133.1,129.8,129.7,129.5,129.4,129.4,129.3,129.0,128.4,128.3,99.1,98.8,95.9,91.0,72.2,71.6,71.5,69.9,69.9,69.3,69.3,68.8,68.5,68.1,67.5,67.5,67.3,67.0,66.6,62.9,62.6,62.4,60.3,57.2,56.8,53.0,52.9,38.6,38.3,27.9,27.8,26.1,25.7,21.0,20.9,20.8,20.7,20.7,20.6,20.5.

HRMS (ESI ^－)[M+HCOO]^－C₄₅H₅₂NO₂₄ theory: 990.2885; experimental 990.2873).

Refining method based on crystallization of 4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero-alpha-D-galacto-non-2-pyron-osyl ] -D-galactopyranose (compound represented by formula G-12)

4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero-alpha-galacto-no-2-pyranosyl ] -D-galactopyranose (a compound represented by formula G-12) (3.00G, 3.17mmol, sialyl alpha/beta ratio = 95.7/4.3) was dissolved in ethyl acetate (4 mL), 2-propanol (60 mL) was added, stirred at 25℃and concentrated to 18mL under reduced pressure. The slurry was stirred at 0℃for 3 hours, and the precipitated crystals were filtered. The filtered crystals were washed with cold 2-propanol (9 mL), dried under reduced pressure at 40 ℃ to give 4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero- α -D-galacto-non-2-pyron-osyl ] -D-galactopyranose (compound represented by formula G-12) (2.66G, yield 88.7%, sialyl α/β ratio = > 99.9/n.d.) as white crystals.

[ Analysis conditions ]

Chromatographic column: CAPCELL PAKADME phi 4.6mm by 150mm, film thickness 3 mu m.

Wavelength: 220nm.

And (3) an oven: 40 ℃.

Eluent: (A) 0.1% aqueous trifluoroacetic acid, (B) acetonitrile.

Gradient: the concentration of the (B) is 40% within 0-150 minutes.

150.1 Min (B) concentration 95%.

155 Minutes (B) concentration 95%.

155.1 Min (B) concentration 40%.

160 Minutes (B) concentration 40%.

Flow rate: 1 mL/min.

And (3) sample injection: 5. Mu.L.

Example 49

4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero-alpha-D-galacto-non-2-pyranosyl ] -1-O- (2, 2-trifluoro-N-phenylethanimido) -D-galactopyranose (Compound of formula D-7)

(Small procedure V-11)

4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero-alpha-D-galacto-non-2-pyranosyl ] -D-galactopyranose (compound of formula G-12) (20.0G, 21.1 mmol) was added to a 500mL eggplant-shaped flask, dichloromethane (200 mL) and molecular sieve 4A powder (10 μm or less, 10.0G) were added, and cooled to 0 ℃. N-methylimidazole (1.91 g,23.3 mmol) and 2, 2-trifluoro-N-phenyliminoacetyl chloride (4.39 g,21.1 mmol) were added under nitrogen at the same temperature, and the mixture was stirred at room temperature for 24 hours. After completion of the reaction was confirmed by HPLC, the reaction mixture was filtered through a filter and washed with methylene chloride (100 mL). The filtrate was filtered through a neutral silica gel pad (silica gel 60N, manufactured by Kanto chemical Co., ltd., particle size: 40 to 50 μm,60 g) filled with methylene chloride, and 100mL each time was taken. The silica gel pad was washed with ethyl acetate/methylene chloride (1:9, 1000mL each time divided, 200 mL), and the selected fraction was concentrated under reduced pressure, whereby 4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero-alpha-D-galacto-non-2-pyron-osyl ] -1-O- (2, 2-trifluoro-N-phenylethanimido) -D-galactopyranose (compound represented by formula D-7) (20.1 g, yield 85%) was obtained as a white amorphous form.

¹H－NMR(500MHz,CDCl₃)δ7.99(dd,2H,J＝8.3,1.4Hz),7.91－7.88(m,2H),7.59－7.56(m,1H),7.51(tt,1H,J＝9.7,2.4Hz),7.44－7.34(m,4H),7.12(t,2H,J＝7.7Hz),7.01(t,1H,J＝7.4Hz),6.82(brs,1H),6.43(brs,2H),5.87－5.77(m,3H),5.52(td,1H,J＝11.0,5.0Hz),5.38－5.34(m,1H),5.18(dd,1H,J＝8.6,1.7Hz),4.95(dd,1H,J＝10.0,2.0Hz),4.53(brs,1H),4.29(dd,1H,J＝12.6,2.9Hz),4.21－4.09(m,3H),4.04(dd,1H,J＝10.0,6.0Hz),3.84(s,3H),3.53(dd,1H,J＝10.3,7.4Hz),2.76(dd,1H,J＝12.9,5.4Hz),2.39(s,3H),2.31(s,3H),2.18(s,3H),2.17(s,3H),2.14(s,3H),2.02(s,3H),1.98(s,3H),1.86(dd,1H,J＝13.0,11.0Hz).

¹³C－NMR(125MHz,CDCl₃)δ174.5,173.5,170.5,170.1,169.8,169.7,169.6,167.2,165.5,165.4,165.3,142.9,133.6,133.3,129.8,129.7,129.6,129.5,129.0,128.7,128.6,128.5,128.4,124.2,119.0,98.7,98.5,70.4,69.8,68.3,68.2,67.5,67.5,66.9,66.7,62.0,61.7,60.3,56.9,53.7,52.9,38.7,31.7,29.2,27.9,26.0,25.8,21.0,21.0,20.9,20.8,20.7,20.6,20.5.

HRMS (ESI ⁺)[M+NH₄]⁺C₅₂H₅₉F₃N₃O₂₂ theory: 1134.3537; experimental 1134.3564).

The spectra were confirmed to be consistent with the following: j.org.chem.2016, 81, 10600-10616.

< Synthesis of Compound represented by formula D-13 >

The compound represented by the formula D-13 was synthesized according to the following synthesis pathway 4.

[ Synthesis path 4]

Example 50

After adding 3,4, 6-tri-O-benzyl-1, 2-O- (1-methoxyethylene) - β -D-mannopyranose (compound represented by formula A-1) (4.56 g,9.00 mmol) to a 200mL eggplant-shaped flask, ethyl acetate (45.6 mL) was added. Water (0.23 mL) and p-TsOH H ₂ O (5.1 mg,0.027 mmol) were added at room temperature under nitrogen atmosphere and stirred at the same temperature for 8 hours. After confirming the completion of the reaction by HPLC, triethylamine (1.25 mL,9.00 mmol) was added thereto and stirred at the same temperature overnight. After completion of rearrangement of acetyl groups by HPLC, 5% sodium bicarbonate water (45 mL) was added to the reaction solution to separate the solution. To the organic layer was added 20% saline (22.8 mL) and the mixture was separated. The organic layer was concentrated to 9mL under reduced pressure, toluene (45.6 mL) was added, and the mixture was concentrated to 9mL in vacuo. Toluene (45.6 mL) was added again and concentrated under reduced pressure to a liquid volume of 9mL. Dehydrated toluene (13.7 mL) was added to obtain a toluene solution of 2-O-acetyl-3, 4, 6-tri-O-benzyl-D-mannopyranose (a compound represented by the formula A-2) as a colorless solution.

Example 51

A toluene solution (9.00 mmol) of 2-O-acetyl-3, 4, 6-tri-O-benzyl-D-mannopyranose (a compound represented by the formula A-2) was charged into a 100mL eggplant-shaped flask, cooled to 0℃and then trichloroacetonitrile (1.95 g,13.5 mmol) and DBU (13.5. Mu.L, 8.96. Mu.mol) were added. Stirred under nitrogen at 0 ℃ for 4 hours. After completion of the reaction was confirmed by HPLC, acetic acid (5.2. Mu.L, 8.96. Mu. Mol) was added to the reaction solution at 0℃to obtain a toluene solution (9.00 mmol) of 2-O-acetyl-3, 4, 6-tri-O-benzyl-1-O- (2, 2-trichloroethane imido) -D-mannopyranose (a compound represented by the formula A-3) as a brown solution. The solution was used directly in the next step.

Example 52

4-Methoxyphenyl 2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-di-oxo-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 6) -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-di-oxo-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt 4) -3, a toluene solution (equivalent to 1.27 mmol) of 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranoside (compound represented by formula A-13) (3.0 g,1.06 mmol) and 2-O-acetyl-3, 4, 6-tri-O-benzyl-1-O- (2, 2-trichloroethane imido) -D-mannopyranose (compound represented by formula A-3) were charged into a 100mL eggplant-shaped flask, toluene (30 mL) and molecular sieve 4A powder (10 μm or less were added, 600 mg). Trimethylsilicone triflate (38.5. Mu.L, 0.212 mmol) was added dropwise under nitrogen at-15℃over 15 minutes and stirred at the same temperature for 1 hour. After completion of the reaction by HPLC, triethylamine (0.19 mL,1.06 mmol) was added and stirred at room temperature for 30 minutes. After the reaction solution was filtered through a filter, the reaction solution was washed with acetonitrile (30 mL), and the obtained filtrate was concentrated under reduced pressure to a liquid volume of 6mL. Acetonitrile (30 mL) was added again and concentrated under reduced pressure to a liquid volume of 6mL. Acetonitrile (30 mL) was added to the concentrated solution, and silica gel 120RP-18 (manufactured by Kanto chemical Co., ltd., particle size: 40 to 50 μm,9.0 g) for reversed phase was added. water (20 mL) was added dropwise over 30 minutes to adsorb the target onto the solid phase, followed by filtration. After washing the solid phase with acetonitrile/water (5:4, 90 mL) (filtrate discard), the target was desorbed with acetonitrile/tetrahydrofuran (9:1, 180 mL). The resulting filtrate was concentrated under reduced pressure to a liquid volume of 6mL. Toluene (30 mL) was added to the resulting solution, and the solution was concentrated under reduced pressure to 6mL in volume. Toluene (30 mL) was added again, and the mixture was concentrated under reduced pressure to a liquid volume of 6mL, whereby a crude product of 4-methoxyphenyl 2-O-acetyl-3, 4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1.fwdarw.3) - [2,3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranosyl- (1.fwdarw.2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1.fwdarw.6) ] -2 was obtained, Toluene solution of 4-di-O-benzyl-beta-D-mannopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranoside (compound represented by formula D-1). this solution was used directly in the next step.

Example 53

4-Methoxyphenyl 2-O-acetyl-3, 4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-di-oxo-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 6) ] -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1- & gt 4) -3, A toluene solution (equivalent to 1.06 mmol) of 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranosyl- (1→4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (a compound represented by formula D-1) was added to a 100mL eggplant-shaped flask, tetrahydrofuran (17.6 mL), methanol (10.5 mL) and methyl trifluoroacetate (0.11 mL, 1.06 mmol) and stirring at 25℃for 10 minutes, potassium tert-butoxide (1 mol/L in tetrahydrofuran) was added (0.53 mL,0.53 mmol). After that, the temperature was raised to 40℃and the mixture was stirred for 2 hours, and then the completion of the reaction was confirmed by HPLC. The reaction solution was cooled to 25℃and acetic acid (0.06 mL,1.03 mmol) and ethyl acetate (35 mL) were added sequentially. The solution was washed twice with 3% saline (35 mL) and once with 20% saline (17.5 mL), and then concentrated under reduced pressure to a liquid volume of 6mL. Toluene (35 mL) was added and concentrated again to a liquid volume of 6mL, toluene (35 mL) was further added and concentrated to a liquid volume of 6mL, and a crude product 4-methoxyphenyl 3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- > 3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- > 2) -3,4 was obtained as a toluene solution, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1.fwdarw.6) ] -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranoside (a compound represented by formula D-2). the product was used directly in the next step.

Example 54

4-Methoxyphenyl 3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-di-oxo-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 6) ] -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, A toluene solution (corresponding to 1.06 mmol) of 3-two oxo-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-two oxo-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranoside (compound represented by formula D-2) and a toluene solution (corresponding to 1.06 mmol) of 4-O-acetyl-3, 6-di-O-benzyl-2-deoxy-2- (1, 3-two oxo-1, 3-dihydro-2H-isoindol-2-yl) -O- [2, 2-trifluoro-N-phenylethanolamido ] -beta-D-glucopyranoside (compound represented by formula D-3) were obtained in quantitative example 39, 1.38mmol equivalent) was added to a 100mL eggplant-shaped flask, and methylene chloride (34.7 mL) and molecular sieve 4A powder (700 mg) were added. T-butyldimethylsilyl triflate (0.12 mL,0.53 mmol) was added dropwise under nitrogen at-78deg.C over 5min and stirred at the same temperature for 9 h. After confirming the completion of the reaction by HPLC, triethylamine (0.15 mL,1.06 mmol) was added thereto and stirred at room temperature for 30 minutes. The reaction solution was filtered through a filter and washed with toluene (30 mL). the obtained filtrate was concentrated under reduced pressure to a liquid amount of 6mL, whereby a crude product of 4-methoxyphenyl 4-acetyl-3, 6-di-O-benzyl-2-deoxy-2- (1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-didanoxy-1 was obtained as a toluene solution, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- > 6) ] -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranoside (a compound represented by formula D-4). the product was used directly in the next step.

Example 55-1

4-Methoxyphenyl 4-acetyl-3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt-2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt-3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt-4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt-2) -3, A toluene solution (equivalent to 1.06 mmol) of 4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1.fwdarw.6) ] -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (a compound represented by formula D-4) was added to a 200mL eggplant-shaped flask, n-butanol (12 mL) and ethylenediamine (12 mL) were added. Stirred under nitrogen at 95℃for 12 hours. After completion of the reaction, the reaction mixture was cooled to room temperature by HPLC, and the reaction mixture was concentrated under reduced pressure to a liquid volume of 6mL. Acetonitrile (40 mL) was added to the concentrated solution, and silica gel 120RP-18 (manufactured by Kanto chemical Co., ltd., particle size: 40 to 50 μm,12.0 g) for reversed phase was added. Water (40 mL) was added dropwise over 30 minutes to adsorb the target onto the solid phase, followed by filtration. After washing the solid phase with acetonitrile/water (3:2, 120 mL) (filtrate discard), the target was desorbed with acetonitrile/tetrahydrofuran (9:1, 180 mL). The resulting filtrate was concentrated under reduced pressure to a liquid volume of 6mL. tetrahydrofuran (40 mL) was further added thereto, and the mixture was concentrated under reduced pressure to a liquid volume of 6mL, whereby a crude product of 4-methoxyphenyl 2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1.fwdarw.2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1.fwdarw.3) - [2,3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl- (1.fwdarw.4) -2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1.fwdarw.2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1.fwdarw.6) ] -2 was obtained, A tetrahydrofuran solution of 4-di-O-benzyl- β -D-mannopyranosyl- (1→4) -2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1→4) -2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranoside (a compound represented by formula D-5). The product was used directly in the next step (example 56).

Example 55-2

The compound represented by formula D-5 was purified by the following method including the step of obtaining fumarate crystals of the compound. By this purification method, the purity of the compound represented by the formula D-5-FMA is greatly improved, and various impurities represented by anomers can be removed into the filtrate.

Crystalline, refined fumaric acid compounds of formula (D-5) based on 4-methoxyphenyl 2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1→2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1→3) - [2,3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl- (1→4) -2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1→2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1→6) ] -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1→4) -2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1→4) -crystalline, FMA-5-to prepare the fumaric acid salt thereof

In a solution of 4-methoxyphenyl 2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 3) - [2,3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl- (1- > 4) -2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 6) ] -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1- > 4) -2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (5 mL) of a compound of the formula (formula D-5 mL) in a solution of the compound shown in the form of ethyl acetate (14 mL) was added dropwise to a solution of the compound shown in the formula (5 mL of the intermediate ethyl acetate at a temperature of 5mL from the theoretical solution of 5 to 10mL (10 mg) at one side of the end of the time of the solution was stirred solution, and the solution was stirred at the end of the solution was not stirred at 25 mL of the end of the solution was 10mL to the solution was prepared. After seed (1 mg) was inoculated and stirred for 1 hour or more, a tetrahydrofuran solution (2 mL) of fumaric acid (82 mg) was added dropwise over 1 hour or more, and the mixture was stirred at 25℃for 30 minutes. Heptane (10 mL) was added dropwise over 1 hour or more, the slurry was filtered, and the resulting crystals were dried under reduced pressure at 25℃to obtain 4-methoxyphenyl 2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1→2) -3,4, 6-tri-O-benzyl- α -D-glucopyranosyl- (1→3) - [2,3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl- (1→4) -2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1→2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1→6) ] -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1→4) -2-amino-3, 6-di-O-benzyl- β -D-galactopyranosyl- (1→4) -2-amino-3, 6-di-O-benzyl- β -D-glucopyranosyl- (1→4) -2, 6-di-O-benzyl- β -D-glucopyranosyl- (1→6%) as white crystals (the intermediate of which was shown in the formula pa, the order of 0.5% (the intermediate of the following: the steps).

[ Analysis conditions ]

Chromatographic column: xselect Fluoro-Phenyl 3.5 μm,4.6 phi. X150mm (Waters).

Wavelength: 220nm.

And (3) an oven: 40 ℃.

Eluent: (A) 10mM NH ₄HCO₃ in water, (B) acetonitrile.

Gradient: the concentration of 0 min (B) was 75%.

25 Minutes (B) concentration 80%.

The concentration of 30 minutes (B) was 100%.

30.01 Min (B) concentration 75%.

35 Min (B) concentration 75%.

Flow rate: 1 mL/min.

And (3) sample injection: 5. Mu.L.

HRMS (ESI ⁺)[M+H]⁺C₁₉₅H₂₁₃N₄O₃₈ ⁺ theory: 3218.4852; experimental 3218.4861).

¹H－NMR(500MHz,CD₃OD)δ7.54(d,J＝7.0Hz,2H),7.48－7.15(m,100H),6.94(d,J＝9.5Hz,2H),6.84(s,10H),5.43(s,1H),5.34(d,J＝9,5Hz,1H),5.25－5.01(m,6H),4.93－4.29(m,40H),4.21－3.43(m,41H),3.42－3.40(m,24H),3.23－2.93(m,6H).

¹³C－NMR(125MHz,CD₃OD)δ155.3,151.4,139.3,139.2,139.06,139.02,138.85,138.83,138.78,138.69,138.5,138.34,133.31,138.19,138.16,138.14,138.07,137.5,128.5,128.45,128.43,128.37,128.34,128.31,128.28,128.26,128.20,128.15,128.10,128.04,127.95,127.86,127.80,127.75,127.72,127.65,127.62,127.59,127.52,127.50,127.4,127.36,127.33,127.31,127.2,126.0,118.6,114.4,103.40,103.36,102.7,102.63,102.59,101.2,99.8,97.9,84.3,83.2,83.1,82.8,82.4,81.6,79.9,78.4,78.2,77.9,77.5,76.1,76.0,75.6,75.5,75.2,75.2,75.1,74.91,74.89,74.8,74.7,74.58,74.55,74.5,74.4,74.3,74.2,74.1,73.9,73.84,73.77,73.4,73.26,73.23,73.1,73.06,72.98,72.93,72.8,72.6,72.5,71.6,71.5,71.3,70.5,69.4,68.79,68.75,68.3,68.2,67.1,57.1,56.4,55.6,55.4.

Example 56

4-Methoxyphenyl 2-amino-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1- & gt 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt 4) -2-amino-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1- & gt 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 6) ] -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1- & gt 4) -2-amino-3, a tetrahydrofuran solution (equivalent to 1.06 mmol) of 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1- > 4) -2-amino-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranoside (a compound represented by formula D-5) was charged into a 100mL eggplant-shaped flask, tetrahydrofuran (17.1 mL) and an aqueous solution of sodium hydrogencarbonate (0.58 g,6.89 mmol) dissolved in water (10.2 mL) were added. Phenyl chloroformate (0.83 g,5.30 mmol) was added and stirred for 1 hour. After completion of the reaction was confirmed by HPLC, ethyl acetate (34 mL) and water (34 mL) were added to conduct a liquid separation operation. The obtained organic layer was concentrated under reduced pressure to a liquid volume of 6mL, acetonitrile (34 mL) was added thereto, and silica gel 120RP-18 (manufactured by Kanto chemical Co., ltd., particle size: 40 to 50 μm,10.2 g) for reverse phase was added. Water (34 mL) was added dropwise over 30 minutes to adsorb the target onto the solid phase, followed by filtration. After washing the solid phase with acetonitrile/water (3:2, 90 mL) (filtrate discard), the target was desorbed with acetonitrile/tetrahydrofuran (9:1, 180 mL). the resulting filtrate was concentrated under reduced pressure to a liquid volume of 6mL. Toluene (34 mL) was further added thereto, and the concentration under reduced pressure was carried out twice to a liquid volume of 6mL, whereby a crude product of 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- [ (phenoxycarbonyl) amino ] -beta-D-glucopyranosyl- (1- & gt 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (phenoxycarbonyl) amino ] -beta-D-glucopyranosyl- (1- & gt 2) -3 was obtained as a toluene solution, 4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1.fwdarw.6) ] -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- [ (phenoxycarbonyl) amino ] - β -D-glucopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- [ (phenoxycarbonyl) amino ] - β -D-glucopyranoside (a compound represented by formula D-6-PHCB). the product was used directly in the next step.

Example 57

4-Methoxyphenyl 4-acetyl-3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt-2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt-3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt-4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt-2) -3, To 4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1.fwdarw.6) ] -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (compound represented by formula D-4) (2.44 g) were added n-butanol (12 mL) and ethylenediamine (12 mL). Stirred under nitrogen at 90℃for 12 hours. After completion of the reaction was confirmed by HPLC, the reaction mixture was cooled to room temperature, and concentrated under reduced pressure. Acetonitrile (50 mL) was added to the residue, and silica gel 120RP-18 (particle size: 40 to 50 μm,7.5g, manufactured by Kato chemical Co., ltd.) was added. Water (50 mL) was added dropwise and the mixture was filtered, and the filtrate was washed with acetonitrile/water (1:1, 200 mL). Thereafter, in order to elute the target substance from the filtrate, acetonitrile (25 mL) and toluene (200 mL) were washed, respectively. The filtrate containing the target was concentrated under reduced pressure. To the resulting residue were added tetrahydrofuran (25 mL) and an aqueous solution of sodium hydrogencarbonate (0.36 g) dissolved in water (12.5 mL). 2, 2-triethyl chloroformate (0.43 mL) was added thereto and stirred for 1 hour. After completion of the reaction was confirmed by HPLC, ethyl acetate (25 mL) and water (25 mL) were added to conduct a liquid separation operation. The obtained organic layer was concentrated under reduced pressure, acetonitrile (25 mL) was added to the residue, and silica gel 120RP-18 (manufactured by Kanto chemical Co., ltd., particle size: 40 to 50 μm,7.5 g) for reverse phase was added. Water (10 mL) was added dropwise to adsorb the target substance to the solid phase, followed by filtration. After washing the filtrate with acetonitrile/water (5:1, 60 mL), the filtrate was washed with toluene (200 mL) to elute the target from the filtrate. The filtrate containing the objective was concentrated under reduced pressure, followed by drying, whereby a crude product of 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- [ (2, 2-trichloroethoxycarbonyl) amino ] - β -D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 3) - [2,3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (2, 2-trichloroethoxy) amino ] - β -D-glucopyranosyl- (1- > 2) -3 was obtained, 2.39g of 4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1.fwdarw.6) ] -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- [ (2, 2-trichloroethoxy) amino ] - β -D-glucopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- [ (2, 2-trichloroethoxy) amino ] - β -D-glucopyranoside (a compound represented by the formula D-6-TROC). the product was used directly in the next step.

Example 58

4-Methoxyphenyl 4-acetyl-3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt-2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt-3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt-4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt-2) -3,4, 6-tri-O-benzyl-alpha-D-pyran A solution of mannosyl- (1.fwdarw.6) ] -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-two oxo-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-two oxo-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (compound of formula D-4) (330 mg,0.088 mmol) in n-butanol (1.65 mL)/ethylenediamine (3.3 mL) was stirred at 90℃for 44 hours. After 2-methyltetrahydrofuran (5.0 mL) was added, the mixture was cooled to room temperature, and water (3 mL) was added. After removal of the aqueous layer, the resulting organic layer was washed three times with 10% methanol water (3 mL) and concentrated to dryness to obtain 4-methoxyphenyl 2-amino-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1.fwdarw.2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1.fwdarw.3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1.fwdarw.4) -2-amino-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1.fwdarw.2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1.fwdarw.6) ] -2, 4-di-O-benzyl-beta-D-glucopyranosyl- (1.fwdarw.4) -2-amino-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1.fwdarw.4) -2-deoxy-beta-D-glucopyranosyl- (1.fwdarw.2-O-benzyl-beta-2-glucopyranosyl- (1.fwdarw.4) -2-tri-O-benzyl-alpha-D-glucopyranosyl- (1.6) of the formula shown in the following.

To the crude product obtained, 4-methoxyphenyl 2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1→2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1→3) - [2,3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl- (1→4) -2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1→2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1→6) ] -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1→4) -2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1→4) -2-deoxy- β -D-glucopyranosyl- (1→2-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1→6-3, 6-di-O-benzyl- α -D-glucopyranosyl- (1→6-6-3, 6-D-methyl-glucopyranoside) shown in the formula (1. 6mL of 3.124 mL, 3M, 3D, 3mL, respectively, and 6mL, 1.31 mmol) and triethylamine (145. Mu.L, 1.56 mmol). After stirring for 6 hours, the crude product obtained by concentrating and drying the reaction mixture was purified by preparative HPLC (acetonitrile/water 93%. Fwdarw.100%), 4-methoxyphenyl 2-acetamido-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- > 3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- > 4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- > 6) ] -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1- > 4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1- > 4) -2-acetamido-2-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1- > 6 mg) is obtained (25 mg of the compound shown in 295.60 mg.

Example 59

To 4-methoxyphenyl 4-acetyl-3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt-4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt 2) room-energy under nitrogen atmosphere 3,4, 6-Tri-O-benzyl- α -D-mannopyranosyl- (1→6) ] -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1→4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranosyl- (1→4) -3, 6-di-O-benzyl-2-deoxy-2- (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (compound of formula D-4) (0.24 g) was added tetrahydrofuran (2 mL) methanol (1 mL), methyl trifluoroacetate (0.04 mL), and 1M potassium tert-butoxide in tetrahydrofuran (0.25 mL). After completion of the reaction was confirmed by HPLC, acetic acid (0.02 mL) was added, and the reaction mixture was concentrated under reduced pressure. Ethyl acetate (2 mL) and water (1 mL) were added to the residue to separate the liquid, and after concentrating the organic layer, the residue was purified by silica gel column chromatography, obtaining 4-methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2-1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2-1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt 2- & gt) galactopyranosyl 0.14g of 3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1.fwdarw.6) ] -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- [1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2-1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranoside (compound represented by formula D-6-PHTH).

Example 60

4-Methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2- [ (phenoxycarbonyl) amino ] - β -D-glucopyranosyl- (1→2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1→3) - [2,3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl- (1→4) -3, 6-di-O-benzyl-2-deoxy-2- [ (phenoxycarbonyl) amino ] - β -D-glucopyranosyl- (1→2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1→6) ] -2 obtained in the above example 56, A toluene solution (equivalent to 1.06 mmol) of 4-di-O-benzyl-beta-D-mannopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (phenoxycarbonyl) amino ] -beta-D-glucopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (phenoxycarbonyl) amino ] -beta-D-glucopyranoside (a compound represented by formula D-6-PHCB) and 4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero-alpha-D-galacto-non-2-pyron yl ] -1-O- (2), 2, 2-trifluoro-N-phenylethanimido) -D-galactopyranoside (the compound represented by formula D-7) (1.78 g,1.59 mmol) was introduced into a 100mL eggplant-shaped flask, and methylene chloride (39.2 mL) and molecular sieve 4A powder (less than 10 μm, 0.78 g) were added. Trimethylsilicone triflate (24.4. Mu.L, 0.11 mmol) was added dropwise under nitrogen at-15℃over 5 minutes and stirred at the same temperature for 1 hour. After completion of the reaction, triethylamine (0.15 mL,1.06 mmol) was added thereto by HPLC, and the mixture was stirred at room temperature for 30 minutes. After the reaction solution was filtered through a filter, the reaction solution was washed with acetonitrile (40 mL), and the obtained filtrate was concentrated under reduced pressure to a liquid volume of 6mL. Acetonitrile (40 mL) was added to the solution, and silica gel 120RP-18 (manufactured by Kanto chemical Co., ltd., particle size: 40 to 50 μm,12.0 g) for reversed phase was added. Water (40 mL) was added dropwise over 30 minutes to adsorb the target onto the solid phase, followed by filtration. after washing the solid phase with acetonitrile/water (3:2, 90 mL) (filtrate discard), the target was desorbed with acetonitrile/tetrahydrofuran (9:1, 180 mL). The obtained filtrate was concentrated to dryness, whereby a crude product 4-methoxyphenyl 4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-O-methyl-D-glycero- α -D-galacto-non-2-pyranosyl- (2.fwdarw.6) -4-O-acetyl-2, 3-di-O-benzoyl- β -D-galactopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- [ (phenoxycarbonyl) amino ] - β -D-glucopyranosyl- (1.fwdarw.2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1.fwdarw.3) - [2, 3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (phenoxycarbonyl) amino ] -beta-D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- > 6) ] -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (phenoxycarbonyl) amino ] -beta-D-glucopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (phenoxycarbonyl) amino ] -beta-D-glucopyranoside (a compound of formula D-8-PHCB) (4.53 g, 92.3% (total yield from the compound represented by formula a-13)). the product was used directly in the next step.

Example 61

3, 6-Di-O-benzyl-2-deoxy-2- [ (2, 2-trichloroethoxycarbonyl) amino ] - β -D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 3) - [2,3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (2, 2-trichloroethoxy) amino ] - β -D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 6) ] -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (2, 2-trichloroethoxy) amino ] -beta-D-glucopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (2, 2-trichloroethoxy) amino ] -beta-D-glucopyranoside (a compound of formula D-6-TROC) (2.37 g) in dichloromethane (25 mL) was added 4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero-alpha-D-galacto-non-2-pyrone glycosyl ] -1-O- (2, 2-trifluoro-N-phenylethanimide) -D-galactopyranose (a compound represented by the formula D-7) (1.01 g) and molecular sieve 4A powder (less than 10 μm, 0.50 g). Triisopropyl silicone triflate (50. Mu.L) was added dropwise at-15℃and stirred at the same temperature for 1 hour. After completion of the reaction by HPLC, triethylamine (84. Mu.L) was added thereto, and the mixture was warmed to room temperature. After the reaction solution was filtered through a filter, the reaction solution was washed with acetonitrile (50 mL), and the obtained filtrate was concentrated under reduced pressure. Refining the residue by reverse phase silica gel chromatography (Biotage SfarBio C D50 g,95% MeCNaq.) gives 4-methoxyphenyl 4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-O-methyl-D-glycero-alpha-D-galacto-non-2-pyron-osyl- (2- > 6) -4-O-acetyl-2, 3-di-O-benzoyl-beta-D-galactopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (2, 2), 2-trichloroethoxycarbonyl) amino ] -beta-D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- > 3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (2, 2-trichloroethoxycarbonyl) amino ] -beta-D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- > 6) ] -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1- > 4) -3, 1.38g of 6-di-O-benzyl-2-deoxy-2- [ (2, 2-trichloroethoxycarbonyl) amino ] - β -D-glucopyranosyl- (1.fwdarw.4) -3, 6-di-O-benzyl-2-deoxy-2- [ (2, 2-trichloroethoxycarbonyl) amino ] - β -D-glucopyranoside (compound represented by formula D-8-TROC).

Example 62

4-Methoxyphenyl 2-acetamido-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- > 3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- > 4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- > 6) ] -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1- > 4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1- & gt 4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranoside (compound of formula D-6-AC) (30 mg, 8.85. Mu. Mol) and 4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycerol-alpha-D-galacto-non-2-pyron-yl ] -1-O- (2, 2-trifluoro-N-phenylethaneimido) -D-galactopyranose (compound of formula D-7) (29.7 mg, to a solution of 0.03mmol in methylene chloride (0.9 mL) was added molecular sieve 4A powder (6 mg,0.2 wt) and the mixture was cooled to 0 ℃. After stirring for 10 minutes, triisopropylsilicone triflate (0.95. Mu.L, 3.54. Mu. Mol) was added. After stirring for 1 hour, t-butyldimethylsilyl triflate (0.81. Mu.L, 3.54. Mu. Mol) was added, and after 1 hour, further t-butyldimethylsilyl triflate (0.81. Mu.L, 3.54. Mu. Mol) was added. After 2 hours, 4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero- α -D-galacto-non-2-pyranosyl ] -1-O- (2, 2-trifluoro-N-phenylethanimide) -D-galactopyranose (compound of formula D-7) (10.0 mg,0.01 mmol) was added and triethylamine (10. Mu.L) was added after stirring for a further 15 hours. From the resulting reaction suspension, the molecular sieve was filtered off at room temperature and washed with dichloromethane (3 mL). The obtained solution was concentrated to dryness, and acetonitrile (1 mL), methanol (0.1 mL) and trifluoroacetic acid (5 μl) were sequentially added. After stirring for 1 hour, triethylamine (10. Mu.L) (reaction solution 1) was added.

In 4-methoxyphenyl 2-acetamido-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1- & gt 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt 4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1- & gt 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 6) ] -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1- & gt 4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1- & gt 4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranoside (compound represented by formula D-6-AC) (206.5 mg,0.06 mmol) and 4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycerol-alpha-D-galacto-non-2-pyroosyl ] -1-O- (2, 2-trifluoro-N-phenylethanimide) -D-galactopyranose (compound represented by formula D-7) (238.3 mg, To a solution of 0.21 mmol) in methylene chloride (6.3 mL) was added molecular sieve 4A powder (20 mg,0.2 wt) and the mixture was cooled to 0 ℃. after stirring for 10 minutes, t-butyldimethylsilyl triflate (7.0 μl,0.03 mmol) was added. After stirring for 24 hours, 4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero-alpha-D-galacto-non-2-pyranosyl ] -1-O- (2, 2-trifluoro-N-phenylethanimido) -D-galactopyranose (compound of formula D-7) (68 mg,0.06 mmol) and tert-butyldimethylsilyl triflate (7.0. Mu.L, 0.03 mmol) were added in sequence, after 7 hours 4-O-acetyl-2 was further added, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero-alpha-D-galacto-non-2-pyranosyl ] -1-O- (2, 2-trifluoro-N-phenylethanimide) -D-galactopyranose (a compound of formula D-7) (68 mg,0.06 mmol) and t-butyldimethylsilyl triflate (7.0. Mu.L, 0.03 mmol). After 14 hours, triethylamine (20. Mu.L) was added, and after molecular sieves were filtered off from the resulting reaction suspension at room temperature, the reaction suspension was washed with dichloromethane (3 mL). The obtained solution was concentrated to dryness, and acetonitrile (4 mL), methanol (0.1 mL) and trifluoroacetic acid (10 μl) were sequentially added. After stirring at 0℃for 1 hour, triethylamine (25. Mu.L) (reaction solution 2) was added.

After mixing the two solutions, the crude product after concentration and drying was purified by reverse phase chromatography (acetonitrile/water 90%. Fwdarw.100%). Concentrating and drying the fraction containing the target substance to obtain a mixture, refining the mixture by silica gel chromatography (ethyl acetate/hexane 50%. Fwdarw.85%), 4-methoxyphenyl 4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-O-methyl-D-glycero-alpha-D-galacto-non-2-pyrone glycosyl- (2- & gt 6) -4-O-acetyl-2, 3-di-O-benzoyl-beta-D-galactopyranosyl- (1- & gt 4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1- & gt 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 3) - [2,3,4, 6-tetra-O-benzyl-beta-D-pyranyl galactosyl- (1.fwdarw.4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1.fwdarw.2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1.fwdarw.6) ] -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1.fwdarw.4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1.fwdarw.4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranoside (a compound represented by formula D-8-AC) (210.5 mg, yield 70%).

Example 63

4-Methoxyphenyl 3, 6-di-O-benzyl-2-deoxy-2-1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt-4) -3, 6-di-O-benzyl-2-deoxy-2-1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) -beta-D-glucopyranosyl- (1- & gt 2) -3, 4, 6-Tri-O-benzyl- α -D-mannopyranosyl- (1→6) ] -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1→4) -3, 6-di-O-benzyl-2-deoxy-2- [1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranosyl- (1→4) -3, 6-di-O-benzyl-2-deoxy-2-1, 3-didanoxy-1, 3-dihydro-2H-isoindol-2-yl) - β -D-glucopyranoside (a compound represented by the formula D-6-PHTH) (14.8 mg) in dichloromethane (0.15 mL), 4-O-acetyl-2, 3-di-O-benzoyl-6-O- [4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-methyl-D-glycero-alpha-D-galacto-non-2-pyranosyl ] -1-O- (2, 2-trifluoro-N-phenylethanimido) -D-galactopyranose (compound of formula D-7) (9.1 mg) and molecular sieve 4A powder (10 μm or less, 3.6 mg) were added. trimethylsilicone triflate (0.25. Mu.L) was added dropwise at-15℃and stirred at the same temperature for 1 hour. After confirming the completion of the reaction by HPLC, triethylamine was added thereto, and the temperature was raised to room temperature. Refining the reaction solution by TLC (toluene-ethyl acetate) to obtain 4-methoxyphenyl 4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-O-methyl-D-glycero-alpha-D-galacto-non-2-pyranosyl- (2- > 6) -4-O-acetyl-2, 3-di-O-benzoyl-beta-D-galactopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) ] -beta-D-glucopyranosyl- (1- > 2) -3 in a white amorphous form, 4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) ] -beta-D-glucopyranosyl- (1- & gt 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 6) ] -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (1- & gt, 7.9mg of 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) ] -beta-D-glucopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (1, 3-dioxido-1, 3-dihydro-2H-isoindol-2-yl) ] -beta-D-glucopyranoside (a compound represented by formula D-8-PHTH).

Example 64

4-Methoxyphenyl 4,7,8, 9-tetra-O-acetyl-3, 5-dideoxy-5- (diacetylamino) -1-O-methyl-D-glycero-alpha-D-galacto-non-2-pyrone glycosyl- (2- & gt 6) -4-O-acetyl-2, 3-di-O-benzoyl-beta-D-galactopyranosyl- (1- & gt 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (phenoxycarbonyl) amino ] -beta-D-glucopyranosyl- (1- & gt-2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt 3) - [2, 3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (phenoxycarbonyl) amino ] -beta-D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- > 6) ] -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (phenoxycarbonyl) amino ] -beta-D-glucopyranosyl- (1- > 4) -3, 6-di-O-benzyl-2-deoxy-2- [ (phenoxycarbonyl) amino ] -beta-D-glucopyranoside (a compound of formula D-8-PHCB) (3.14 g, 0.678 mmol) was placed in a 100mL eggplant-shaped flask, 1, 2-dimethoxyethane (28.3 mL), water (9.4 mL) and an aqueous lithium hydroxide solution (4M, 3.39mL,13.6 mmol) were added in this order, and the mixture was stirred for 1 hour. After completion of the reaction, by HPLC, acetic acid (0.19 mL,4.89 mmol) was added. Then, when water (31 mL) was added, a gel-like solid precipitated. The resulting solid was filtered, and the solid phase was washed with 1, 2-dimethoxyethane/water (9:10, 31 mL) to give the crude product 4-methoxyphenyl 5-acetamido-3, 5-dideoxy-D-glycero- α -D-galacto-non-2-galactopyranosyl- (2- > 6) - β -D-galactopyranosyl- (1- > 4) -2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 3) - [2,3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl- (1- > 4) -2-amino-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1.fwdarw.2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1.fwdarw.6) ] -2, 4-di-O-benzyl-beta-D-mannopyranosyl- (1.fwdarw.4) -2-amino-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1.fwdarw.4) -2-amino-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranoside (a compound represented by formula D-9). the product was used directly in the next step.

The HPLC purity of the compounds represented by formula D-9 obtained by removing the protecting group of the amino group of the compounds represented by formula D-8 obtained in examples 61 to 63 under the deprotection reaction conditions shown in the following table is also shown in the table. As shown in the table, in particular, when phenoxycarbonyl (COOPh) was used as a protecting group for an amino group, deprotection was carried out at room temperature within 1 hour under normal hydrolysis conditions, and the series of steps of glycosylation, deprotection, and further acetylation of an amino group were carried out at the most favorable yield without decomposing the substrate.

TABLE 6

Example 65

4-Methoxyphenyl 5-acetamido-3, 5-dideoxy-D-glycero- α -D-galacto-non-2-galactopyranosyl- (2- > 6) - β -D-galactopyranosyl- (1- > 4) -2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 3) - [2,3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl- (1- > 4) -2-amino-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1- > 2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1- > 6) ] -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1- > 4) -2-amino-3, 6-di-O- β -D-mannopyranosyl- (1- > 4) -2-amino-3, 6-D-galactopyranosyl- (1- > 6-D-glucopyranosyl- (1- > 6-D-glucopyranosyl- (2- > 6) is added to a wet solid of the formula (678 mmol) shown in a flask, tetrahydrofuran (18.8 mL), methanol (9.4 mL) and triethylamine (3.31 mL,23.7 mmol) were added sequentially. Acetic anhydride (1.59 mL,17.0 mmol) was then added and stirred at room temperature for 2 hours. After confirming the completion of the reaction by HPLC, water (31.4 mL) and ethyl acetate (62.8 mL) were added to the reaction solution, and the obtained organic layer was concentrated and dried to give a crude product 4-methoxyphenyl 5-acetamido-3, 5-dideoxy-D-glycero- α -D-galacto-non-2-pyrone glycosyl- (2.fwdarw.6) - β -D-galactopyranosyl- (1.fwdarw.4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1.fwdarw.2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1.fwdarw.3) - [2,3,4, 6-tetra-O-benzyl- β -D-galactopyranosyl- (1.fwdarw.4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1.fwdarw.2) -3, 6-tri-O-benzyl- α -D-mannopyranose- (1.fwdarw.2) -3, 6-di-benzyl- β -D-glucopyranosyl- (1.fwdarw.2) -3, 6-O-benzyl- β -D-mannopyranose compound represented by the formula (1.fwdarw.2) -3-2-g.6-tri-benzyl-O-D-mannopyranose).

Example 66

In 4-methoxyphenyl 5-acetamido-3, 5-dideoxy-D-glycero-alpha-D-galacto-non-2-pyrone glycosyl- (2- & gt, 6) -beta-D-galactopyranosyl- (1- & gt, 4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy-beta-D-glucopyranosyl- (1- & gt, 2) -3,4, 6-tri-O-benzyl-alpha-D-mannopyranosyl- (1- & gt, 3) - [2,3,4, 6-tetra-O-benzyl-beta-D-galactopyranosyl- (1- & gt, 4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy-beta-D-pyranyl to a mixture of glucosyl- (1.fwdarw.2) -3,4, 6-tri-O-benzyl- α -D-mannopyranosyl- (1.fwdarw.6) ] -2, 4-di-O-benzyl- β -D-mannopyranosyl- (1.fwdarw.4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranosyl- (1.fwdarw.4) -2-acetamido-3, 6-di-O-benzyl-2-deoxy- β -D-glucopyranoside (compound of formula D-10) (corresponding to 0.678 mmol) was added N-methylpyrrolidone (26.1 mL) and 5% Pd/C (Chuan, EA type, 54.9% aqueous solution, 3.92 g), and repeating the pressure reduction and nitrogen substitution three times. After that, hydrogen pressurization and decompression are repeated three times. Stirring was carried out at an external temperature of 50℃and a hydrogen pressure of 0.5MPa for 48 hours. After completion of the reaction was confirmed by HPLC, the reaction solution was filtered under reduced pressure in a glove box substituted with nitrogen, and the filtrate was washed with N-methylpyrrolidone (35 mL). Toluene (45 mL) was added to the filtrate, and the mixture was concentrated to 55mL. This operation was repeated three times, and a dehydration operation was performed, whereby a crude product of 4-methoxyphenyl 5-acetamido-3, 5-dideoxy-D-glycero- α -D-galacto-non-2-pyrone glycosyl- (2→6) - β -D-galactopyranosyl- (1→4) -2-acetamido-2-deoxy- β -D-glucopyranosyl- (1→2) - α -D-mannopyranosyl- (1→3) - [ β -D-galactopyranosyl- (1→4) -2-acetamido-2-deoxy- β -D-glucopyranosyl- (1→2) - α -D-mannopyranosyl- (1→6) ] - β -D-mannopyranosyl- (1→4) -2-acetamido-2-deoxy- β -D-glucopyranosyl- (1→4) -2-acetamido-deoxy- β -D-glucopyranoside (a compound represented by formula D-11) was obtained as an N-methylpyrrolidone solution. The product was used directly in the next step. The catalyst had almost the same reactivity even when 5% Pd/C (developed in Chuan, PE type) or ASCA-2 (developed in N.E. CHEMCAT) was used instead of the above 5% Pd/C (developed in Chuan, EA type).

Example 67

The compound represented by D-12 was purified according to the following synthesis route W. The compounds shown below under the left are examples of compounds in which "R ₇" in formula E1 is methyl.

[ Synthesis path W ]

Example 68

11-Azido-3, 6, 9-trioxaundecan-1-amine (-) -di-p-toluoyl-L-tartrate (Compound (R ₇ =Me) represented by formula E-2)

To a solution of 11-azido-3, 6, 9-trioxaundecan-1-amine (compound of formula D-12) (10.0 g,45.82mmol,94.3% HPLC purity) in acetonitrile (24 mL) and water (6 mL) was added (-) -di-p-toluoyl-L-tartaric acid (17.70 g,45.82 mmol), and the mixture was stirred at 35℃to confirm dissolution, and acetonitrile (300 mL) was added dropwise over 1 hour at the same temperature. The resulting slurry was concentrated to 200mL under reduced pressure, and the slurry was stirred at 25℃for 30 minutes, and the precipitated crystals were filtered. The filtered crystals were washed with acetonitrile (30 mL) and dried under reduced pressure at 40 ℃ to give 11-azido-3, 6, 9-trioxaundecan-1-amine (-) -di-p-toluoyl-L-tartrate (compound of formula E-2-pta) (24.40 g, yield 88.0%,99.2% hplc purity) (melting point 152 ℃) as white crystals.

¹H－NMR(500MHz,DMSO)δ7.82(d,J＝9.0Hz,4H),7.30(d,J＝9.0Hz,4H),5.61(s,2H),3.59(t,J＝5.0Hz,2H),3.56－3.47(m,10H),3.39(t,J＝5.5Hz,2H),2.91(t,J＝6.0Hz,2H),2.36(s,6H).

¹³C－NMR(125MHz,DMSO)δ168.06,164.80,143.63,129.25,129.18,126.93,71.68,69.72,69.63,69.59,69.57,69.20,66.68,49.96,38.40,21.14.

The same procedure was carried out even if (+) -di-p-toluoyl-D-tartaric acid was used instead of (-) -di-p-toluoyl-L-tartaric acid.

Example 69

11-Azido-3, 6, 9-trioxaundec-1-amine (-) -dibenzoyl-L-tartrate (a compound represented by formula E-2-BZTA)

To a solution of 11-azido-3, 6, 9-trioxaundecan-1-amine (compound represented by formula D-12) (5.0 g,22.91 mmol) in acetonitrile (12 mL) and water (3 mL) was added (-) -dibenzoyl-L-tartaric acid (8.21 g,22.91 mmol), and after stirring at 35℃and confirming dissolution, acetonitrile (150 mL) was added dropwise over 1 hour at the same temperature. The resulting slurry was concentrated to 125mL under reduced pressure, acetonitrile (100 mL) was added, and concentrated again to 125mL under reduced pressure. The slurry was stirred at 25℃for 30 minutes and the precipitated crystals were filtered. The filtered crystals were washed with acetonitrile (15 mL) and dried under reduced pressure at 40 ℃ to give 11-azido-3, 6, 9-trioxaundecan-1-amine (-) -dibenzoyl-L-tartrate salt (compound represented by formula E-2-BZTA) (10.6 g, yield 80%) as white crystals (melting point 131 ℃).

¹H－NMR(500MHz,DMSO)δ7.94(d,J＝8.5Hz,4H),7.64(t,J＝7.5Hz,2H),7.51(t,J＝7.5Hz,4H),5.65(s,2H),3.59(t,J＝5.0Hz,2H),3.56－3.47(m,10H),3.39(t,J＝5.5Hz,2H),2.90(t,J＝5.0Hz,2H).

¹³C－NMR(125MHz,DMSO)δ167.98,164.86,133.38,129.63,129.22,128.66,72.10,69.73,69.63,69.59,69.57,69.21,66.69,49.97,38.38.

The same procedure was carried out even if (+) -dibenzoyl-D-tartaric acid was used instead of (-) -dibenzoyl-L-tartaric acid.

Example 70

11-Azido-3, 6, 9-trioxaundec-1-amine (-) -di-p-methoxybenzoyl-L-tartrate (Compound (R ₇ =OMe) represented by formula E-2)

To a solution of 11-azido-3, 6, 9-trioxaundecan-1-amine (compound represented by formula D-12) (5.0 g,22.91 mmol) in acetonitrile (12 mL) and water (3 mL) was added (-) -di-p-methoxybenzoyl-L-tartaric acid (9.58 g,22.91 mmol), and after stirring at 35℃and confirming dissolution, acetonitrile (150 mL) was added dropwise over 1 hour at the same temperature. The resulting slurry was concentrated to 100mL under reduced pressure, the slurry was stirred at 25℃for 30 minutes, and the precipitated crystals were filtered. The filtered crystals were washed with acetonitrile (15 mL), dried under reduced pressure at 40 ℃ to give 11-azido-3, 6, 9-trioxaundecan-1-amine (-) -di-p-methoxybenzoyl-L-tartrate salt (compound represented by formula E-2-PATA (R ₇ =ome)) as white crystals (11.4 g, yield 78%) (melting point 153 ℃).

¹H－NMR(500MHz,DMSO)δ7.89(d,J＝9.0Hz,4H),7.02(d,J＝9.0Hz,4H),5.59(s,2H),3.82(s,6H),3.59(t,J＝5.0Hz,2H),3.56－3.47(m,10H),3.39(t,J＝5.5Hz,2H),2.91(t,J＝5.5Hz,2H).

¹³C－NMR(125MHz,DMSO)δ168.25,164.50,163.14,131.34,121.88,113.92,71.64,69.73,69.63,69.59,69.21,66.70,55.48,49.97,38.38.

The same procedure can be performed even if (+) -di-p-methoxybenzoyl-D-tartaric acid is used instead of (-) -di-p-methoxybenzoyl-L-tartaric acid.

Example 71

11-Azido-3, 6, 9-trioxaundecan-1-amine (Compound represented by formula D-12)

To a solution of 11-azido-3, 6, 9-trioxaundecan-1-amine (-) -di-p-toluoyl-L-tartrate (compound of formula E-2-PTTA) (12.0 g,19.85 mmol) obtained in example 68 in ethyl acetate (120 mL) and water (18 mL) was added concentrated hydrochloric acid (2.41 g,23.82 mmol), and the mixture was stirred at 25℃and then separated. The aqueous layer was washed twice with ethyl acetate (120 mL), and after adjusting the pH to 11 with 10N aqueous sodium hydroxide (2.15 mL,21.50 mmol), sodium chloride (0.6 g) was added to dissolve. Dichloromethane (120 mL) was added thereto and stirred, followed by separation to obtain an organic layer. Further, methylene chloride (120 mL) was added to the aqueous layer and stirred, followed by separation, and the resulting organic layers were combined. The combined organic layers were concentrated to 12mL under reduced pressure, acetonitrile (120 mL) was added, and concentrated to 12mL under reduced pressure. The resulting solution was filtered, washed with acetonitrile (6 mL) after removing inorganic salts, and the resulting solution was concentrated under reduced pressure to give 11-azido-3, 6, 9-trioxaundecan-1-amine (compound represented by formula D-12) with an HPLC purity of 99.2% as a pale yellow oily compound (3.7 g, yield 85%).

¹H－NMR(500MHz,CDCl₃)δ3.69－3.62(m,10H),3.51(t,J＝5.5Hz,2H),3.39(t,J＝5.5Hz,2H),2.87(t,J＝5.5Hz,2H).

< Purity analysis conditions of Compounds represented by formula D-12 >

The purity of the compound represented by formula D-12 was measured under the following HPLC analysis conditions. After measuring the sample solution, a blank solution was measured, and the blank peak was removed, thereby calculating the HPLC purity of the compound represented by formula D-12.

< Preparation of sample solution >

50Mg of the compound represented by the formula D-12 was weighed into a 10mL volumetric flask, and 2mL of acetonitrile and 100. Mu.L of triethylamine were added thereto to mix them. Acetic anhydride 50. Mu.L was added thereto, and the mixture was left standing for 15 minutes after mixing, to acetyl-derivatize the compound represented by the formula D-12. 150. Mu.L of 4N aqueous sodium hydroxide solution was added, and after mixing, a sample solution was prepared by constant volume with 50% acetonitrile water.

Preparation of blank solution

2ML of acetonitrile and 100. Mu.L of triethylamine were added to a 10mL volumetric flask and mixed. Acetic anhydride 50. Mu.L was added thereto, and the mixture was left to stand for 15 minutes after mixing. 150. Mu.L of 4N aqueous sodium hydroxide solution was added, and after mixing, a sample solution was prepared by constant volume with 50% acetonitrile water.

< HPLC analysis conditions >)

The using device comprises: SHIMAZU HPLC (LC-20 AD).

Chromatographic column: xbrige C18.3.5 μm,4.6 mm. Times.150 mm (Waters).

Mobile phase a:10mM AcONH ₄ in water.

Mobile phase B: CH ₃ CN.

TABLE 7

Gradient conditions:

Flow rate: 1 mL/min.

Detection wavelength: 210nm.

Column temperature: 40 ℃.

Injection amount: 5. Mu.L.

Retention time: 7.3 minutes (acetylated compound of formula D-12), 9.0 minutes (acetylated dimer of compound of formula D-12).

Example 72

4-Methoxyphenyl 5-acetamido-3, 5-dideoxy-D-glycero- α -D-galacto-non-2-pyron-osyl- (2→6) - β -D-galactopyranosyl- (1→4) -2-acetamido-2-deoxy- β -D-glucopyranosyl- (1→2) - α -D-mannopyranosyl- (1→3) - [ β -D-galactopyranosyl- (1→4) -2-acetamido-2-deoxy- β -D-glucopyranosyl- (1→2) - α -D-mannopyranosyl- (1→6) ] - β -D-mannopyranosyl- (1→4) -2-acetamido-2-deoxy- β -D-glucopyranoside (compound represented by formula D-11) (corresponding to 0.678 mmol) N-methylpyrrolidone solution was added to 100mL eggplant-shaped flask, 11-azido-2-D-mannopyranosyl- (1→4) - β -D-mannopyranosyl- (1→6) ] - β -D-mannopyranosyl- (1→6) - β -D-mannopyranosyl- (1→4) -2-acetamido-D-galactopyranosyl- (1→2.032.032-6) ] -acetamido-D-glucopyranosyl- (1→4) -6-D-galactopyranosyl- (1.3.3-6-triazolyl- (3.3 mmol) and 3.3-undecyl-6-triazolyl-N-yl-3-N-D-glucopyranosyl (3.3 mmol) were added at the same time. After completion of the reaction, by HPLC, when acetonitrile (300 mL) was added dropwise to the reaction solution, a gel-like solid precipitated. The suspension was centrifuged. The white gel-like solid was washed twice with acetonitrile (30 mL), water (10 mL) was added to the obtained white gel-like solid, and the mixture was filtered through a membrane filter, and the filtrate was purified by HPLC (loading per time: 2 mL). The fraction containing the target was concentrated under reduced pressure and then freeze-dried, whereby 4-methoxyphenyl N- (2- {2- [2- (2-azidoethoxy) ethoxy ] ethoxy } ethyl) -5-acetyl-ceramide- (2→6) - β -D-galactopyranosyl- (1→4) -2-acetamido-2-deoxy- β -D-glucopyranosyl- (1→2) - α -D-mannopyranosyl- (1→3) - [ β -D-galactopyranosyl- (1→4) -2-acetamido-2-deoxy- β -D-glucopyranosyl- (1→2) - α -D-mannopyranosyl- (1→6) ] - β -D-mannopyranosyl- (1→4) -2-acetamido-2-deoxy- β -D-glucopyranosyl- (1→4) -2-deoxy- β -D-glucopyranoside (compound represented by formula D-13) (1.01 g, 66% (total purity of the compound represented by D8 (detection of 98.2 nm)), was obtained as a white solid.

¹H－NMR(500MHz,D₂O)δ1.73(t,J＝12.0Hz,1H),1.87－2.07(brs,15H),2.59(dd,J＝4,12.0Hz,1H),3.26－3.90(m,77H),4.00(brs,1H),4.08(brs,1H),4.14(brs,1H),4.30－4.38(m,2H),4.50－4.54(m,3H),4.65(brs,1H),4.78(d,J＝5.5Hz,1H),4.83(brs,1H),4.93(d,J＝8.5Hz,1H),5.03(brs,1H),6.86(d,J＝9.5Hz,2H),6.93(d,J＝9.5Hz,2H).

¹³C－NMR(125MHz,D₂O)δ22.71,22.76,22.89,22.99,23.06,23.91,38.85,39.41,50.80,52.35,55.22,55.50,55.62,56.45,60.58,60.62,60.84,61.64,62.28,62.34,63.29,63.68,66.31,66.43,67.63,67.91,67.92,67.99,68.45,68.97,69.14,69.18,69.80,69.85,70.06,70.10,70.16,70.28,70.83,71.29,71.62,71.73,72.66,72.71,72.73,73.09,73.16,73.50,74.12,74.16,74.19,75.02,75.05,75.34,75.36,75.99,76.98,77.06,79.19,79.65,80.10,81.11,81.48,97.67,100.02,100.10,100.18,101.07,102.03,103.60,104.29,115.72,118.92,151.66,155.49,163.48,169.72,175.26,175.28,175.39,175.46,175.64.

HRMS (ESI) [ m+h ] ⁺ (M/z): theoretical value C ₈₈H₁₄₄N₉O₅₇: 2238.8641; experimental value 2238.8604[ m+h ] ⁺.

Claims

1. A method for producing an oligosaccharide represented by the following formula A-13,

The method comprises a working procedure I-1, a working procedure I-2 and a working procedure I-3,

The step I-1 is a step of producing a compound represented by the following formula A-7, and includes the steps of: by forming an alpha-1, 6-glycosidic bond between a compound represented by the formula A-3 and a compound represented by the following formula A-4 to produce a compound represented by the following formula A-5,

The step I-2 is a step of producing a compound represented by the following formula A-10, and includes the steps of: by forming a beta-1, 4-glycosidic bond between the compound represented by the formula A-7 and the compound represented by the following formula A-8 to produce a compound represented by the following formula A-9,

The step I-3 is a step of producing the oligosaccharide represented by the formula A-13, and includes the steps of: by forming a beta-1, 2-glycosidic bond between the compound represented by the formula A-10 and the compound represented by the formula A-11 below, a compound represented by the formula A-12 below is produced,

2. The method of claim 1, wherein,

The process I-2 includes: the compound represented by the formula A-10 is produced by reacting the compound represented by the formula A-9 with a strong base in the presence of an alkyl ester of a perfluorocarboxylic acid.

3. The method of claim 2, wherein,

The alkyl ester of perfluorocarboxylic acid is methyl trifluoroacetate, ethyl trifluoroacetate, propyl trifluoroacetate, isopropyl trifluoroacetate, butyl trifluoroacetate, methyl pentafluoropropionate, ethyl pentafluoropropionate, propyl pentafluoropropionate, isopropyl pentafluoropropionate, methyl heptafluorobutyrate, ethyl heptafluorobutyrate, propyl heptafluorobutyrate, isopropyl heptafluorobutyrate, butyl heptafluorobutyrate, methyl nonafluoropentanoate, ethyl nonafluoropentanoate, propyl nonafluoropentanoate, isopropyl nonafluoropentanoate, butyl nonafluoropentanoate, methyl undecanoate, ethyl undecanoate, propyl undecanoate, isopropyl undecanoate, or butyl undecanoate.

4. A method according to claim 2 or 3, wherein,

The strong base is selected from the group consisting of: sodium, lithium, and potassium salts of metal amides; sodium, lithium, potassium, cesium, and barium salts of C1-C20 alkoxides; sodium hydride, potassium hydride, lithium hydride, butyllithium, potassium carbonate, sodium carbonate, cesium carbonate, lithium carbonate, potassium phosphate, sodium phosphate, cesium phosphate, lithium phosphate, diazabicycloundecene DBU, diazabicyclononene DBN, 1, 3-tetramethylguanidine TMG; and combinations thereof.

5. A method according to claim 2 or 3, wherein,

The strong base is potassium tert-butoxide, sodium tert-butoxide, lithium tert-butoxide or lithium hexamethyldisilazide LHMDS.

6. The method according to any one of claims 2 to 5, wherein,

The reaction of step I-2 is carried out in a C1-C10 alcohol solvent alone or in a mixture of a C1-C10 alcohol solvent and an amide solvent, an ether solvent, an ester solvent, an aromatic solvent, a halogen solvent, a hydrocarbon solvent, or a nitrile solvent.

7. The method according to any one of claims 1 to 6, wherein,

The process I-3 includes: the compound represented by the formula A-12 is reacted with 2, 3-dichloro-5, 6-dicyano-p-benzoquinone DDQ in a mixed solvent of a fluoroalcohol and water to thereby release the 2-naphthylmethyl group in the compound represented by the formula A-12, thereby producing the oligosaccharide represented by the formula A-13.

8. The method of claim 7, wherein,

The fluoroalcohol is selected from the group consisting of hexafluoro-2-propanol HFIP, 2-trifluoroethanol TFE 2,3, 4, 5-octafluoro-1-pentanol, nonafluoro-tert-butanol, and combinations thereof.

9. The method according to claim 7 or 8, wherein,

The reaction of the procedure I-3 is carried out at the temperature of minus 35 ℃ to 70 ℃.

10. The method according to claim 7 or 8, wherein,

The reaction of the procedure I-3 is carried out at the temperature of minus 30 ℃ to minus 10 ℃.

11. The method according to any one of claims 1 to 10, wherein,

The step I-1 includes: after stopping the reaction between the compound represented by the formula A-4 and the compound represented by the formula A-3, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the produced compound represented by the formula A-5 and inclusions, the compound represented by the formula A-5 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and the water, whereby the inclusions are removed, and then the compound represented by the formula A-5 is eluted from the hydrophobic carrier using an organic solvent, whereby the compound represented by the formula A-5 is purified.

12. The method according to any one of claims 1 to 11, wherein,

The process I-2 includes: after stopping the reaction between the compound represented by the formula A-7 and the compound represented by the formula A-8, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the produced compound represented by the formula A-9 and inclusions, the compound represented by the formula A-9 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and the water, whereby the inclusions are removed, and then the compound represented by the formula A-9 is eluted from the hydrophobic carrier using an organic solvent, whereby the compound represented by the formula A-9 is purified.

13. The method according to any one of claims 1 to 12, wherein,

The process I-3 includes: after stopping the reaction between the compound represented by the formula A-10 and the compound represented by the formula A-11, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the produced compound represented by the formula A-12 and inclusions, the compound represented by the formula A-12 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and the water, whereby the inclusions are removed, and then the compound represented by the formula A-12 is eluted from the hydrophobic carrier using an organic solvent, whereby the compound represented by the formula A-12 is purified.

14. The method of claim 11, wherein,

The inclusion contains a sugar compound other than the compound represented by the formula A-5 and/or a compound derived from a reactant for obtaining the purified compound.

15. The method of claim 12, wherein,

The inclusion contains a sugar compound other than the compound represented by the formula A-9 and/or a compound derived from a reactant for obtaining the purified compound.

16. The method of claim 13, wherein,

The inclusion contains a sugar compound other than the compound represented by the formula A-12 and/or a compound derived from a reactant for obtaining the purified compound.

17. The method according to any one of claims 11 to 16, wherein,

The hydrophobic carrier is a resin for filling by reversed phase partition chromatography.

18. The method of claim 17, wherein,

The reverse phase partition chromatography packing resin is selected from the group consisting of poly (styrene/divinylbenzene) polymer gel resins, polystyrene-divinylbenzene resins, polyhydroxymethacrylate resins, styrene-vinylbenzene copolymer resins, polyvinyl alcohol resins, polystyrene resins, polymethacrylate resins, chemically bonded silica gel resins, and combinations thereof.

19. The method of claim 18, wherein,

The chemically bonded silica gel resin is selected from the group consisting of: (1) A resin obtained by reacting a silane coupling agent with silica gel; (2) A resin obtained by chemically bonding dimethyloctadecyl, octadecyl, trimethyloctadecyl, dimethyloctyl, octyl, butyl, ethyl, methyl, phenyl, cyanopropyl, or aminopropyl to silica gel; (3) A resin obtained by chemically bonding a behenyl group or a triacontyl group to silica gel; and (4) combinations of the above (1) to (3).

20. The method of claim 18, wherein,

The chemical bonding silica gel resin is octadecyl bonding silica gel resin, namely ODS resin.

21. The method according to any one of claims 11 to 20, wherein,

The water-soluble organic solvent is a water-soluble alcohol solvent, a water-soluble nitrile solvent, a water-soluble ether solvent, a water-soluble ketone solvent, a water-soluble amide solvent, or a water-soluble sulfoxide solvent, or a mixed solvent comprising at least one of the above water-soluble organic solvents.

22. The method of claim 21, wherein,

The water-soluble nitrile solvent is acetonitrile.

23. The method according to any one of claims 11 to 22, wherein,

The organic solvent used in the step of eluting the target from the hydrophobic carrier may be a nitrile solvent, an ether solvent, an ester solvent, a ketone solvent, a halogen solvent, an aromatic solvent, or a mixed solvent comprising at least one of the solvents.

24. The method according to any one of claims 1 to 23, wherein,

The compound represented by the formula A-11 is produced by a process comprising a process Y-1 and a process Y-2,

The step Y-1 is a step of producing a compound represented by the following formula B-4, and includes the steps of: by forming a beta-1, 4-glycosidic bond between a compound represented by the following formula B-1 and a compound represented by the following formula B-2 to produce a compound represented by the following formula B-3,

The process Y-2 is as follows: adding lithium t-butoxide or lithium t-amyl alcohol to a solvent comprising the compound represented by the formula B-4 and benzyl halide or benzyl sulfonate, protecting the hydroxyl group existing in the compound represented by the formula B-4 with benzyl, thereby generating a compound represented by the following formula B-5,

25. The method of claim 24, wherein,

The solvent comprising the compound represented by the formula B-4 and benzyl halide or benzyl sulfonate is an amide solvent, an ether solvent, an aromatic solvent, a hydrocarbon solvent, a urea solvent, or a mixed solvent comprising at least one of the above solvents.

26. The method of claim 24 or 25, wherein,

The method also comprises the following steps: opening the ring of the phthalimide group in the compound represented by the formula B-5, then generating a crystalline compound represented by the following formula B-6 by reacting with cinchonidine, separating the crystalline compound represented by the formula B-6 from an amorphous substance, removing cinchonidine in the compound represented by the formula B-6 by adding an acidic aqueous solution and a solvent to generate a compound represented by the following formula B-7, then closing the ring of the ring-opened phthalimide group in the compound represented by the formula B-7, thereby refining the compound represented by the formula B-5,

27. The method according to any one of claims 1 to 26, wherein,

The method also comprises the following steps: opening the phthalimide group in the compound represented by the formula A-13, then forming a salt with (R) - (+) -1- (1-naphthyl) ethylamine to produce a crystalline compound represented by the following formula A-14, separating the crystalline compound represented by the formula A-14 from an amorphous substance, removing the (R) - (+) -1- (1-naphthyl) ethylamine in the compound represented by the formula A-14 by adding an acidic aqueous solution and a solvent to produce a compound represented by the following formula A-15, then closing the ring of the phthalimide group ring in the compound represented by the formula A-15 by the ring opening, thereby purifying the compound represented by the formula A-13,

28. A method for producing an oligosaccharide represented by the following formula D-13,

The method comprises a working procedure II-1, a working procedure II-2, a working procedure II-3 and a working procedure II-4,

The step II-1 is a step of producing a compound represented by the following formula D-2, and includes the steps of: by forming an alpha-1, 3-glycosidic bond between an oligosaccharide represented by the following formula A-13 and a compound represented by the following formula A-3 to produce a compound represented by the following formula D-1,

The procedure II-2 is as follows: after the compound represented by the following formula D-5 is produced, the amino group in the compound represented by the formula D-5 is protected with a protecting group selected from the group consisting of aryloxycarbonyl COOAr, acetyl Ac, 2-trichloroethoxycarbonyl Troc, and phthalimide group Pht to produce a compound represented by the following formula D-6, wherein R ₅ is aryloxycarbonyl COOAr, acetyl Ac, or 2, 2-trichloroethoxycarbonyl Troc, and R ₆ is a hydrogen atom, or R ₅ and R ₆ form a phthalimide group together with the nitrogen atom to which they are bonded; or alternatively

Removing the acetyl Ac on the compound represented by the following formula D-4 to produce a compound represented by the following formula D-6, wherein R ₅ and R ₆ together with the nitrogen atom to which they are bonded form a phthalimide group,

The step II-2 includes a step of forming a beta-1, 2-glycosidic bond between the compound represented by the formula D-2 and the compound represented by the formula D-3 to produce a compound represented by the formula D-4,

The step II-3 is a step of producing a compound represented by the following formula D-11, and includes the steps of: forming a beta-1, 4-glycosidic bond between the compound represented by the formula D-6 and the compound represented by the formula D-7 to form a compound represented by the formula D-8, removing the protecting group of the amino group and the acyl protecting group of the alcohol on the compound represented by the formula D-8 to form a compound represented by the formula D-9,

In the formula D-8, R ₅ is aryloxycarbonyl COOAr, acetyl Ac, or 2, 2-trichloroethoxycarbonyl Troc, and R ₆ is a hydrogen atom, or R ₅ and R ₆ form a phthalimide group together with the nitrogen atom to which they are bonded,

In the formula D-9, M ⁺ is sodium ion, lithium ion, potassium ion or triethylamine cation which is protonated,

In the formula D-11, M ⁺ is sodium ion, lithium ion, potassium ion or triethylamine cation which is protonated,

The procedure II-4 is as follows: reacting the compound represented by the formula D-11 with a compound represented by the following formula D-12 to produce the oligosaccharide represented by the formula D-13,

29. The method of claim 28, wherein,

The process II-1 includes: the compound represented by the formula D-2 is produced by reacting the compound represented by the formula D-1 with a strong base in the presence of an alkyl ester of a perfluorocarboxylic acid.

30. The method of claim 29, wherein,

31. The method of claim 29 or 30, wherein,

32. The method of claim 29 or 30, wherein,

33. The method according to any one of claims 29 to 32, wherein,

The step of producing the compound represented by the formula D-2 by reacting the compound represented by the formula D-1 with a strong base in the presence of trifluoroacetate is performed in a C1-C10 alcohol solvent alone or in a mixed solvent of a C1-C10 alcohol solvent and an amide-based solvent, an ether-based solvent, an ester-based solvent, an aromatic-based solvent, a halogen-based solvent, a hydrocarbon-based solvent, or a nitrile-based solvent.

34. The method according to any one of claims 28 to 33, wherein,

In the step II-3, the compound represented by the formula D-6 is produced by protecting the amino group in the compound represented by the formula D-5 with an aryloxycarbonyl COOAr.

35. The method according to any one of claims 28 to 34, wherein,

In the step II-3, the step of producing the compound represented by the formula D-6 from the compound represented by the formula D-5 is performed in an aqueous solution of sodium hydrogencarbonate, potassium hydrogencarbonate, disodium hydrogenphosphate, or dipotassium hydrogenphosphate.

36. The method according to any one of claims 28 to 35, wherein,

The compound represented by the formula D-12 is obtained by a purification method comprising the steps of:

a step of adding a compound represented by the following formula E-1 to a solution containing the crude compound represented by the formula D-12 to produce a crystalline compound represented by the following formula E-2; and

Isolating the crystalline compound, followed by a step of extracting the compound represented by the formula D-12 from the isolated crystalline compound,

In the formula E-1, R ₇ is a hydrogen atom, a methyl group or a methoxy group,

In formula E-2, R ₇ is a hydrogen atom, a methyl group, or a methoxy group.

37. The method of claim 36, wherein,

The purified compound represented by the formula D-12 has a purity of 95% or more as measured by HPLC.

38. The method of claim 37, wherein,

The purity is above 98%.

39. The method according to any one of claims 28 to 38, wherein,

The process II-1 includes: after stopping the reaction between the compound represented by the formula A-13 and the compound represented by the formula A-3, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the produced compound represented by the formula D-1 and inclusions, the compound represented by the formula D-1 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and the water, whereby the inclusions are removed, and then the compound represented by the formula D-1 is eluted from the hydrophobic carrier using an organic solvent, whereby the compound represented by the formula D-1 is purified.

40. The method of any one of claims 28-39, wherein,

The process II-2 includes: after stopping the reaction between the compound represented by the formula D-3 and the compound represented by the formula D-4, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the compound represented by the formula D-5 and inclusions formed, the compound represented by the formula D-5 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and the water, whereby the inclusions are removed, and then the compound represented by the formula D-5 is eluted from the hydrophobic carrier using an organic solvent, whereby the compound represented by the formula D-5 is purified.

41. The method of any one of claims 28 to 40, wherein,

The process II-3 includes: after stopping the reaction between the compound represented by the formula D-6 and the compound represented by the formula D-7, a hydrophobic carrier and water are added to a water-soluble organic solvent containing the produced compound represented by the formula D-8 and inclusions, the compound represented by the formula D-8 is adsorbed on the hydrophobic carrier, and then the hydrophobic carrier is filtered and washed with a mixed solution of the water-soluble organic solvent and the water, whereby the inclusions are removed, and then the compound represented by the formula D-8 is eluted from the hydrophobic carrier using an organic solvent, whereby the compound represented by the formula D-8 is purified.

42. The method of any one of claims 28 to 41, wherein,

The process II-3 includes: the compound represented by the formula D-10 is purified by forming a compound represented by the following formula D-10 by protecting an amino group on the formula D-9 with an acetyl group, adding a hydrophobic carrier and water to a water-soluble organic solvent containing the formed compound represented by the formula D-10 and impurities, adsorbing the compound represented by the formula D-10 to the hydrophobic carrier, filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and the water, thereby removing the impurities, eluting the compound represented by the formula D-10 from the hydrophobic carrier with an organic solvent,

43. The method of claim 39, wherein,

The inclusion contains a sugar compound other than the compound represented by the formula D-1, and/or a compound derived from a reactant for obtaining the purified compound.

44. The method of claim 40, wherein,

The inclusion contains a sugar compound other than the compound represented by the formula D-5, and/or a compound derived from a reactant for obtaining the purified compound.

45. The method of claim 41, wherein,

The inclusion contains a sugar compound other than the compound represented by the formula D-8, and/or a compound derived from a reactant for obtaining the purified compound.

46. The method of claim 42, wherein,

The inclusion contains a sugar compound other than the compound represented by the formula C-10, and/or a compound derived from a reactant for obtaining the purified compound.

47. The method of any one of claims 39-46, wherein,

48. The method of claim 47, wherein,

49. The method of claim 48, wherein,

50. The method of claim 48, wherein,

51. The method of any one of claims 39-50, wherein,

52. The method of claim 51, wherein,

The water-soluble nitrile solvent is acetonitrile.

53. The method of any one of claims 39-52, wherein,

54. The method of any one of claims 28-53, wherein,

The method also comprises the following steps: forming a salt of the compound represented by the formula D-5 with fumaric acid to produce a crystalline compound represented by the following formula D-5-FMA, separating and purifying the crystalline compound represented by the formula D-5-FMA from an amorphous substance,

55. A method for producing a compound represented by the following formula B-5, which comprises the steps of: adding lithium tert-butoxide or lithium tert-amyl alcohol to a solvent comprising a compound represented by the following formula B-4 and benzyl halide or benzyl sulfonate, protecting a hydroxyl group existing in the compound represented by the formula B-4 with benzyl,

56. The method of claim 55, wherein,

The solvent is an amide solvent, an ether solvent, an aromatic solvent, a urea solvent, a hydrocarbon solvent, or a mixed solvent containing at least one of the above solvents.

57. A method for producing a compound represented by the following formula a-10, which comprises the steps of: reacting a compound represented by the following formula A-9 with a strong base in the presence of an alkyl ester of a perfluorocarboxylic acid,

58. The method of claim 57, wherein,

59. The method of claim 57 or 58, wherein,

The strong base is selected from: sodium, lithium, potassium salts of metal amides; sodium, lithium, potassium, cesium, and barium salts of C1-C20 alkoxides; sodium hydride, potassium hydride, lithium hydride, butyllithium, potassium carbonate, sodium carbonate, cesium carbonate, lithium carbonate, potassium phosphate, sodium phosphate, cesium phosphate, lithium phosphate, diazabicycloundecene DBU, diazabicyclononene DBN, 1, 3-tetramethylguanidine TMG; and combinations thereof.

60. The method of claim 57 or 58, wherein,

61. The method of any one of claims 57-60, wherein,

The reaction is carried out in a C1-C10 alcohol solvent alone or in a mixed solvent of a C1-C10 alcohol solvent and an amide-based solvent, an ether-based solvent, an ester-based solvent, an aromatic-based solvent, a halogen-based solvent, a hydrocarbon-based solvent, or a nitrile-based solvent.

62. A method for producing an oligosaccharide represented by the following formula A-13,

The method comprises the following steps: reacting a compound represented by the following formula A-12 with 2, 3-dichloro-5, 6-dicyano-p-benzoquinone DDQ in a mixed solvent of a fluoroalcohol and water to release 2-naphthylmethyl group in the compound represented by the formula A-12,

63. The method of claim 62, wherein,

64. The method of claim 62 or 63, wherein,

At a temperature of between-35 and 70 ℃.

65. The method of claim 62 or 63, wherein,

At a temperature of between-30 ℃ and-10 ℃.

66. A method for purifying a compound represented by the following formula a-5, comprising:

adding a hydrophobic carrier and water to a water-soluble organic solvent containing the compound represented by the formula a-5 and inclusions, allowing the compound represented by the formula a-5 to adsorb in the hydrophobic carrier, then filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and the water, thereby removing the inclusions, and then eluting the compound represented by the formula a-5 from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula a-5.

67. A method for purifying a compound represented by the following formula a-9, comprising:

Adding a hydrophobic carrier and water to a water-soluble organic solvent containing the compound represented by the formula a-9 and inclusions, allowing the compound represented by the formula a-9 to adsorb in the hydrophobic carrier, then filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and the water, thereby removing the inclusions, and then eluting the compound represented by the formula a-9 from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula a-9.

68. A method for purifying a compound represented by the following formula a-12, comprising:

adding a hydrophobic carrier and water to a water-soluble organic solvent containing the compound represented by the formula a-12 and inclusions, allowing the compound represented by the formula a-12 to adsorb in the hydrophobic carrier, then filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and the water, thereby removing the inclusions, and then eluting the compound represented by the formula a-12 from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula a-12.

69. A method for purifying a compound represented by the following formula D-1, which comprises:

Adding a hydrophobic carrier and water to a water-soluble organic solvent containing the compound represented by the formula D-1 and inclusions, allowing the compound represented by the formula D-1 to adsorb to the hydrophobic carrier, then filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and the water, thereby removing the inclusions, and then eluting the compound represented by the formula D-1 from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula D-1.

70. A method for purifying a compound represented by the following formula D-5, which comprises:

adding a hydrophobic carrier and water to a water-soluble organic solvent containing the compound represented by the formula D-5 and inclusions, allowing the compound represented by the formula D-5 to adsorb in the hydrophobic carrier, then filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and the water, thereby removing the inclusions, and then eluting the compound represented by the formula D-5 from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula D-5.

71. A method for purifying a compound represented by the following formula D-8,

The method comprises the following steps: adding a hydrophobic carrier and water to a water-soluble organic solvent containing the compound represented by the formula D-8 and inclusions, allowing the compound represented by the formula D-8 to adsorb in the hydrophobic carrier, then filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and the water, thereby removing the inclusions, and then eluting the compound represented by the formula D-8 from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula D-8.

72. A method for purifying a compound represented by the following formula D-10, which comprises:

Adding a hydrophobic carrier and water to a water-soluble organic solvent containing the compound represented by the formula D-10 and inclusions, allowing the compound represented by the formula D-10 to adsorb in the hydrophobic carrier, then filtering and washing the hydrophobic carrier with a mixed solution of the water-soluble organic solvent and the water, thereby removing the inclusions, and then eluting the compound represented by the formula D-10 from the hydrophobic carrier using an organic solvent, thereby purifying the compound represented by the formula D-10.

73. The method of claim 69, wherein,

74. The method of claim 70, wherein,

The inclusion contains a sugar compound other than the compound shown in D-5, and/or a compound derived from a reactant for obtaining the purified compound.

75. The method of claim 71, wherein,

The inclusion contains a sugar compound other than the compound represented by D-8, and/or a compound derived from a reactant for obtaining the purified compound.

76. The method of claim 72, wherein,

The inclusion contains a sugar compound other than the compound represented by the formula D-10, and/or a compound derived from a reactant for obtaining the purified compound.

77. The method of any one of claims 69 to 76 wherein,

78. The method of claim 77, wherein,

79. The method of claim 78, wherein,

80. The method of claim 79, wherein,

81. The method of any one of claims 69-80 wherein,

The water-soluble organic solvent is a water-soluble alcohol solvent, a water-soluble nitrile solvent, a water-soluble ether solvent, a water-soluble ketone solvent, a water-soluble amide solvent, a water-soluble sulfoxide solvent, or a mixed solvent comprising at least one of the above water-soluble organic solvents.

82. The method of claim 81, wherein,

The water-soluble nitrile solvent is acetonitrile.

83. The method of any one of claims 69-82 wherein,

84. A method for producing a compound represented by the following formula D-8, wherein,

The method comprises the following steps: generating a compound represented by the following formula D-6, then forming a beta-1, 4-glycosidic bond between the compound represented by the formula D-6 and a compound represented by the following formula D-7, thereby generating a compound represented by the formula D-8,

In formula D-6, R ₅ is aryloxycarbonyl COOAr, acetyl Ac, or 2, 2-trichloroethoxycarbonyl Troc, and R ₆ is a hydrogen atom, or R ₅ and R ₆ together with the nitrogen atom to which they are bonded form a phthalimide group,

85. The method of claim 84, wherein,

R ₅ is aryloxycarbonyl COOAr.

86. The method of claim 84 or 85, comprising:

removing the protecting group of the amino group and the acyl protecting group of the alcohol in the compound shown in the formula D-8 to generate a compound shown in the formula D-9,

Wherein M ⁺ is sodium ion, lithium ion, potassium ion, or protonated triethylamine cation.

87. A method for purifying a compound represented by the following formula D-12, which comprises the steps of:

In formula E-2, R ₇ is a hydrogen atom, a methyl group, or a methoxy group.

88. An oligosaccharide represented by the following formula A-13,

89. A compound represented by the following formula A-5,

90. A compound represented by the following formula A-6,

91. A compound represented by the following formula A-7,

92. A compound represented by the following formula A-9,

93. A compound represented by the following formula A-10,

94. A compound represented by the following formula A-11,

95. A compound represented by the following formula A-12,

96. A compound represented by the following formula A-14,

97. A compound represented by the following formula A-15,

98. A compound represented by the following formula B-4,

99. A compound represented by the following formula B-5,

100. A compound represented by the following formula B-6,

101. A compound represented by the following formula B-7,

102. A compound represented by the following formula B-8,

103. A compound represented by the following formula D-1,

104. A compound represented by the following formula D-2,

105. A compound represented by the following formula D-4,

106. A compound represented by the following formula D-5,

107. A compound represented by the following formula D-5-FMA,

108. A compound represented by the following formula D-6,

Wherein R ₅ is an aryloxycarbonyl COOAr, acetyl Ac, or 2, 2-trichloroethoxycarbonyl Troc, and R ₆ is a hydrogen atom, or R ₅ and R ₆ together with the nitrogen atom to which they are bonded form a phthalimide group.

109. A compound represented by the following formula D-8,

110. A compound represented by the following formula D-9,

111. A compound represented by the following formula D-10,

112. A compound represented by the following formula D-11,

113. A crystalline compound represented by the following formula E-2,

Wherein R ₇ is a hydrogen atom, a methyl group, or a methoxy group.

114. A compound represented by the following formula D-12, wherein,

Has a purity of 90% or more when measured by HPLC,

115. The compound of claim 109, wherein,

The purity is more than 95%.

116. An oligosaccharide represented by the following formula D-13,