Glycan, and method for producing medicine containing glycan

Abstract

To provide a novel method for pure-chemically producing a diantennary branched N-glycan having an α2,6-sialic acid structure at a non-reducing terminal thereof with excellent yield, selectivity, and efficiency; a method for producing a novel monosaccharide or oligosaccharide useful in the method, an intermediate for the production, and others; and a novel method for producing a glycoprotein utilizing the method.SOLUTION: Provided are a novel method for pure-chemically producing a diantennary branched N-glycan having an α2,6-sialic acid structure at a non-reducing terminal thereof, in which a synthesis scheme such that two sugar blocks are produced separately and then the two sugar blocks are linked to a polysaccharide block stereoselectively is utilized for a glycoside bond that has been difficult to construct directly utilizing the involvement of an adjacent group; a method for producing a novel monosaccharide or oligosaccharide useful in the method, an intermediate for the production, and others; and a novel method for producing a glycoprotein utilizing the method.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a novel purely chemical method for producing biantennary N-glycans having an α2,6-sialic acid structure at the non-reducing end, a novel method for producing monosaccharides or oligosaccharides and intermediates useful in the purely chemical method for producing the N-glycans, and a novel method for producing the N-glycans including the production method, and a novel method for producing glycoproteins and the like (in particular, a glycan-remodeling antibody or its FC region-containing molecule, or an antibody-drug conjugate) using the novel purely chemical method for producing the N-glycans. [Background technology]

[0002] Glycosylation of proteins is known to have a significant impact on their function and structure. N-linked glycans, in particular, are deeply involved in the physiological activity of proteins, and it has been reported that biantennary N-glycans with an α2,6-sialic acid structure at the non-reducing end are the optimal structure for enhancing antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) (Non-Patent Document 1).

[0003] In order to develop and commercialize pharmaceuticals that utilize glycans, it is desirable to be able to stably produce high-purity glycans in large quantities at an industrially affordable price. The synthesis of α2,6-sialylglycans that has been reported so far can be broadly divided into two methods: 1) semi-chemical synthesis by purification of natural extracts and chemical enzymatic conversion, and 2) pure chemical synthesis.

[0004] For example, it has been reported that N-linked glycans can be obtained from chicken egg yolk by combining an enzymatic method and a chemical method as a semi-chemical synthesis method (Non-Patent Document 2). While these methods can synthesize the target glycans with fewer steps than pure chemical synthesis, they require the procurement of large amounts of egg yolk, and in many cases, special techniques and purification equipment are required for the subsequent isolation and purification from the egg yolk and for the purification of the water-soluble unprotected glycans after chemical conversion (Patent Documents 1 to 4).

[0005] On the other hand, the following examples have been reported as purely chemical synthesis methods for α2,6-sialylglycan. (1) Total synthesis of a complex 11-saccharide glycan containing an α2,6-sialyl moiety (Non-patent Document 3) (2) Total synthesis of immunoglobulin G13 glycopeptide having an α2,6-sialyl moiety (Non-patent Document 4) (3) Total synthesis of α2,6-sialyl12-saccharide N-linked glycan containing core fucose (Non-patent Document 5) (4) Total synthesis of α2,6-sialyldecasaccharide N-linked sugar chains fluorinated at position 3 (Non-patent document 6)

[0006] If a robust production method is established for purely chemical synthesis of glycans, it is expected that the degree of freedom in production volume will be extremely high by deriving from monosaccharides, just like ordinary low molecular weight compounds. Furthermore, since the conversion of sugars modified with protecting groups is involved, most of the purification procedures involve handling water-insoluble compounds, and it is expected that the complexity and labor required will be significantly reduced compared to semi-chemical synthesis methods.

[0007] On the other hand, the previously reported examples mentioned above pose two major challenges in synthesis: 1) the highly difficult conversion of sugar parts, such as the construction of β-mannoside parts and α-sialyl parts, and the linking process thereof, involve low-selectivity, low-yield steps; and 2) in both the sugar part conversion and linking processes, silica gel column chromatographic purification, which is not suitable for scale-up, is frequently used in each step, and precise preparative chromatographic purification procedures are essential in many steps to remove isomers and impurities generated as by-products in the reaction.

[0008] As described above, purely chemical synthesis of glycans has potential advantages over semi-chemical synthesis when it comes to large-scale synthesis. However, it cannot be said that technological development has progressed sufficiently in terms of yield, selectivity, efficiency, and cost, and there are very few examples of this method being adopted for actual large-scale synthesis. [Prior art documents] [Patent documents]

[0009] [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 [Non-patent literature]

[0010] [Non-Patent Document 1] Proc.Natl.Acad.Sci.USA2015,112,10611 [Non-patent document 2] BeilsteinJ.Org.Chem.2018,14,416 [Non-patent document 3] TetrahedronLett.1986,27,5739 [Non-patent document 4] J.Am.Chem.Soc.2009,131,16669 [Non-patent document 5] J.Org.Chem.2016,81,10600 [Non-patent document 6] J.Am.Chem.Soc.2019,141,6484 Summary of the Invention [Problem to be solved by the invention]

[0011] One objective of the present invention is to provide a novel purely chemical method for producing biantennary N-glycans having an α2,6-sialic acid structure at the non-reducing end, which is excellent in yield, selectivity, and efficiency. Another objective of the present invention is to provide a novel method for producing monosaccharides or oligosaccharides or intermediates useful in the purely chemical production method for the N-glycans, as well as a novel method for producing the N-glycans including the production method. Yet another objective of the present invention is to provide a novel method for producing glycoproteins and the like (particularly, glycochain-remodeling antibodies or their FC region-containing molecules, or antibody-drug conjugates) using the novel purely chemical production method for N-glycans, etc. [Means for solving the problem]

[0012] As a result of intensive studies to solve the above-mentioned problems, the present inventors have discovered a novel purely chemical method for producing biantennary N-glycans having an α2,6-sialic acid structure at the non-reducing end, using a synthetic scheme in which glycosidic bonds that are difficult to construct directly using neighboring group participation are individually generated as disaccharide blocks and then stereoselectively linked to other saccharide blocks. Furthermore, they have discovered novel methods for producing monosaccharides or oligosaccharides and intermediates useful in the purely chemical production method for said N-glycans, novel methods for producing said N-glycans that include said production methods, and novel methods for producing glycoproteins, etc. (particularly, glycan-remodeling antibodies or their FC region-containing molecules, or antibody-drug conjugates) using the novel purely chemical production method for said N-glycans, etc., thereby completing the present invention.

[0013] That is, the present invention relates to, but is not limited to, the following:

[0014] [1] Formula XI below: [ka] A method for producing an oligosaccharide represented by the formula: (Step 1) Formula III-3 below: [ka] with a compound providing a leaving group selected from the group consisting of a trifluoromethanesulfonyloxy group, a nonafluorobutanesulfonyloxy group, a 2-nitrobenzenesulfonyloxy group, and a 4-nitrobenzenesulfonyloxy group, to obtain a compound of the following formula III-4: [ka] (wherein X1 represents a substituent selected from the group consisting of a trifluoromethanesulfonyl group, a nonafluorobutanesulfonyl group, a 2-nitrobenzenesulfonyl group, and a 4-nitrobenzenesulfonyl group), and then the compound represented by formula III-4 is reacted with cesium acetate or tetrabutylammonium acetate to give a compound represented by formula III-5: [ka] wherein X2 is an acetyl group, or by reacting a compound of formula III-4 with tetrabutylammonium benzoate to give a compound of formula III-5: [ka] wherein X2 is a benzoyl group, to form a compound represented by the following formula III-11: [ka] or a compound of formula III-13: [ka] a step of preparing a compound represented by (Step 2) Reacting a compound represented by formula III-13 with a compound represented by formula IV-3: [ka] By reacting a compound represented by the following formula V-1: [ka] The method of claim 1, further comprising the step of producing a compound represented by formula V-3: [ka] or a process for producing a compound represented by A compound represented by formula III-11 and the following formula IV-3: [ka] By reacting a compound represented by the following formula V-4: [ka] The method of claim 1, further comprising the step of producing a compound represented by formula V-5: [ka] a step of preparing a compound represented by (Step 3) Reacting a compound represented by formula V-3 with a compound represented by formula II-6: [ka] to obtain a compound represented by the following formula VI-1: [ka] The method of claim 1, further comprising the step of producing a compound of formula VI-3: [ka] or a process for preparing a compound represented by formula V-5 and a compound represented by formula II-9: [ka] to give a compound of formula VI-4: [ka] The method of claim 1, further comprising the step of producing a compound of formula VI-3: [ka] or a compound of formula VI-6: [ka] a step of preparing a compound represented by (Step 4) Formula VIII-5 below: [ka] and a compound represented by the following formula VII-3: [ka] is bonded to a compound represented by the following formula IX-1: [ka] The method of claim 1, further comprising the step of producing a compound of formula IX-5: [ka] a process for producing a compound represented by the formula: (Step 5) A compound represented by formula VI-3 is reacted with a compound represented by formula IX-5 to obtain a compound represented by formula X-1: [ka] or The compound of formula VI-6 is reacted with the compound of formula IX-5 to give the compound of formula X-3: [ka] a process for producing an oligosaccharide of formula XI, comprising the step of forming a compound of formula XI A method comprising: [2] Step 1 is to prepare a compound of formula III-1: [ka] is reacted with a strong alkoxide base in the presence of methyl trifluoroacetate to give a compound of the following formula III-2: [ka] The method according to [1], comprising a step of producing a compound represented by the formula: [3] The method according to [2], wherein the alkoxide-based strong base is selected from the group consisting of sodium salts, lithium salts, potassium salts of C1 to C5 alkoxides, and combinations thereof. [4] Formula XI below: [ka] A method for producing an oligosaccharide represented by the formula: (Step 1) Formula III-3 below: [ka] In the compound represented by the formula III-7, the 2-position of the D-glucopyranoside is oxidized to obtain a compound represented by the formula III-7: [ka] Then, in the compound of formula III-7, the oxo group attached to the carbon atom at position 2 of the 2-keto-D-glucopyranoside is reduced to give a compound of formula III-8: [ka] The method of claim 1, further comprising the step of forming a compound of formula III-11: [ka] or a compound of formula III-13: [ka] a step of preparing a compound represented by (Step 2) Reacting a compound represented by formula III-13 with a compound represented by formula IV-3: [ka] By reacting a compound represented by the following formula V-1: [ka] The method of claim 1, further comprising the step of producing a compound represented by formula V-3: [ka] or a process for producing a compound represented by A compound represented by formula III-11 and the following formula IV-3: [ka] By reacting a compound represented by the following formula V-4: [ka] The method of claim 1, further comprising the step of producing a compound represented by formula V-5: [ka] a step of preparing a compound represented by (Step 3) Reacting a compound represented by formula V-3 with a compound represented by formula II-6: [ka] to obtain a compound represented by the following formula VI-1: [ka] The method of claim 1, further comprising the step of producing a compound of formula VI-3: [ka] or a process for preparing a compound represented by formula V-5 and a compound represented by formula II-9: [ka] to give a compound of formula VI-4: [ka] The method of claim 1, further comprising the step of producing a compound of formula VI-3: [ka] or a compound of formula VI-6: [ka] a step of preparing a compound represented by (Step 4) Formula VIII-5 below: [ka] and a compound represented by the following formula VII-3: [ka] is bonded to a compound represented by the following formula IX-1: [ka] The method of claim 1, further comprising the step of producing a compound of formula IX-5: [ka] a process for producing a compound represented by the formula: (Step 5) A compound represented by formula VI-3 is reacted with a compound represented by formula IX-5 to obtain a compound represented by formula X-1: [ka] or The compound of formula VI-6 is reacted with the compound of formula IX-5 to give the compound of formula X-3: [ka] a process for producing an oligosaccharide of formula XI, comprising the step of forming a compound of formula XI A method comprising: [5] Step 1 is to prepare a compound of formula III-1: [ka] is reacted with a strong alkoxide base in the presence of methyl trifluoroacetate to give a compound of the following formula III-2: [ka] The method according to [4], comprising a step of producing a compound represented by the formula: [6] The method according to [5], wherein the alkoxide-based strong base is selected from the group consisting of sodium salts, lithium salts, potassium salts of C1 to C5 alkoxides, and combinations thereof. [7] In step 1, L-Selectride, LS-Selectride, lithium diisobutyl-tert-butoxyaluminum hydride (LDBBA), a compound of formula A: [ka] wherein R3 is a group represented by the following formula: [ka] The method according to any one of [4] to [6], wherein the compound represented by formula III-7 is reduced in the presence of a reducing agent selected from the group consisting of compounds represented by formula III-7 and combinations thereof, wherein at least two R3 are di-tert-butylmethylphenoxides or hydrides represented by formula III-8, [8] In step 1, a compound of formula A below is used: [ka] wherein R3 is a group represented by the following formula: [ka] The method according to any one of [4] to [7], wherein the compound represented by formula III-7 is reduced in the presence of a reducing agent which is a compound represented by the following formula: [9] Step 1 is to prepare a compound of formula III-9: [ka] The method further comprises the step of preparing a compound represented by formula III-9 by reacting a compound represented by formula III-8 in the presence of lithium tert-butoxide or lithium tert-amoxide: [ka] The method according to any one of [1] to [8], comprising a step of protecting the hydroxyl group bonded to the carbon atom at position 2 of the D-mannopyranoside in a compound represented by formula III-9 with a benzyl group to produce a compound represented by formula III-9.

[10] Step 2 is a compound of formula V-5: [ka] and the step of preparing the compound of formula V-5 further comprises a step of preparing a compound of formula V-4: [ka] The method according to any one of [1] to [9], comprising a step of reacting a compound represented by formula V-1 with a strong alkoxide base in the presence of methyl trifluoroacetate to produce a compound represented by formula V-5.

[11] The method according to

[10] , wherein the alkoxide-based strong base is selected from the group consisting of sodium salts, lithium salts, potassium salts of C1 to C5 alkoxides, and combinations thereof.

[12] Step 3 further comprises the step of preparing a compound of formula II-6 or a compound of formula II-9; The process for preparing the compound of formula II-6 comprises reacting a compound of formula II-4 in a fluorous alcohol and water: [ka] With λ3-iodane, a compound represented by the following formula II-5: [ka] producing a compound represented by the formula: The process for preparing the compound of formula II-9 comprises reacting a compound of formula II-7 in a fluorous alcohol and water: [ka] The compound represented by the formula II-8 can be obtained by reacting the compound represented by the formula [ka] The method according to any one of [1] to

[11] , comprising a step of producing a compound represented by the following formula:

[13] 12. The method of claim 11, wherein the λ3-iodane is selected from the group consisting of [bis(trifluoroacetoxy)iodo]benzene (PIFA), [hydroxy(tosyloxy)iodo]benzene (HTIB), (diacetoxyiodo)benzene (PIDA), [bis(trifluoroacetoxy)iodo]pentafluorobenzene, [hydroxy(methanesulfonyloxy)iodo]benzene, and combinations thereof.

[14] The method according to

[12] or

[13] , wherein the fluorous alcohol is selected from the group consisting of hexafluoro-2-propanol (HFIP), 2,2,2-trifluoroethanol (TFE), 2,2,3,3,4,4,5,5-octafluoro-1-pentanol, nonafluoro-tert-butyl alcohol, and combinations thereof.

[15] Step 3 further comprises the step of preparing a compound of formula II-6 or a compound of formula II-9; The process for preparing the compound of formula II-6 comprises reacting a compound of formula II-5: [ka] in the presence of N-methylimidazole, with a compound of the formula: [ka] with 2,2,2-trifluoro-N-phenylacetimidoyl chloride (TFPC) of formula II-6, The process for preparing the compound of formula II-9 comprises reacting a compound of formula II-8: [ka] in the presence of N-methylimidazole, with a compound of the formula: [ka] The method according to any one of [1] to

[14] , comprising a step of reacting a compound represented by formula II-9 with 2,2,2-trifluoro-N-phenylacetimidoyl chloride (TFPC) represented by the formula:

[16] Step 3 is a compound of formula VI-5: [ka] The process for preparing the compound of formula VI-5 further comprises a step of preparing a compound of formula VI-4: [ka] The method according to any one of [1] to

[15] , comprising a step of reacting a compound represented by the formula: with a strong alkoxide base in the presence of methyl trifluoroacetate to produce a compound represented by the formula VI-5.

[17] The method according to

[16] , wherein the alkoxide-based strong base is selected from the group consisting of sodium salts, lithium salts, potassium salts of C1 to C5 alkoxides, and combinations thereof.

[18] The method according to any one of [1] to

[17] , wherein step 4 further comprises a step of producing a compound represented by formula VII-3, and the step of producing the compound represented by formula VII-3 comprises a step of contacting a solvent in which the compound represented by formula VII-3 is dissolved with silica gel to perform solid-phase extraction of the compound represented by formula VII-3.

[19] The method according to

[18] , wherein the solvent in which the compound represented by formula VII-3 is dissolved is selected from the group consisting of toluene, dichloromethane, chloroform, and combinations thereof.

[20] Step 4 is to prepare a compound of formula VIII-3: [ka] in the presence of N-methylimidazole, with a compound of the formula: [ka] By reacting with 2,2,2-trifluoro-N-phenylacetimidoyl chloride (TFPC) represented by the following formula VIII-4: [ka] The method according to any one of [1] to

[19] , comprising a step of producing a compound represented by the following formula: [twenty one] Step 4 is to prepare a compound of formula IX-3: [ka] The method according to any one of [1] to

[20] , further comprising a step of producing a compound represented by formula IX-3, wherein the step of producing the compound represented by formula IX-3 comprises a step of contacting a solvent in which the compound represented by formula IX-3 is dissolved with silica gel to perform solid-phase extraction of the compound represented by formula IX-3. [twenty two] The method according to

[21] , wherein the solvent in which the compound represented by formula IX-3 is dissolved is selected from the group consisting of toluene, dichloromethane, chloroform, and combinations thereof. [twenty three] Formula III-2 below: [ka] A method for preparing a compound represented by the following formula III-1: [ka] with a strong base of alkoxide type in the presence of methyl trifluoroacetate to produce a compound of formula III-2. [twenty four] The method according to

[23] , wherein the alkoxide-based strong base is selected from the group consisting of sodium salts, lithium salts, potassium salts of C1 to C5 alkoxides, and combinations thereof. [twenty five] Formula III-5 below: [ka] (wherein X2 is an acetyl group), the method for producing a compound represented by the following formula III-4: [ka] (wherein X1 represents a substituent selected from the group consisting of trifluoromethanesulfonyl, nonafluorobutanesulfonyl, 2-nitrobenzenesulfonyl, and 4-nitrobenzenesulfonyl) with cesium acetate or tetrabutylammonium acetate to produce a compound of formula III-5.

[26] Formula III-5 below: [ka] (wherein X2 is a benzoyl group), the method for producing a compound represented by the following formula III-4: [ka] wherein X1 represents a substituent selected from the group consisting of trifluoromethanesulfonyl, nonafluorobutanesulfonyl, 2-nitrobenzenesulfonyl, and 4-nitrobenzenesulfonyl, with tetrabutylammonium benzoate to produce a compound of formula III-5.

[27] Formula III-8 below: [ka] A method for preparing a compound represented by the following formula III-7: [ka] by reducing the oxo group attached to the carbon atom at position 2 of the 2-keto-D-glucopyranoside of the compound represented by formula III-8 to produce a compound represented by formula III-9.

[28] L-Selectride, LS-Selectride, lithium diisobutyl-tert-butoxyaluminum hydride (LDBBA), represented by the following formula A: [ka] wherein R3 is a group represented by the following formula: [ka] The method according to

[27] , wherein the oxo group bonded to the carbon atom at the 2-position of the 2-keto-D-glucopyranoside in the compound of formula III-7 is reduced in the presence of a reducing agent selected from the group consisting of compounds represented by formula III-7:

[29] Formula A below: [ka] wherein R3 is a group represented by the following formula: [ka] The method according to

[27] or

[28] , wherein the oxo group bonded to the carbon atom at the 2-position of the 2-keto-D-glucopyranoside in the compound of formula III-7 is reduced in the presence of a reducing agent, which is a compound of formula III-7 (di-tert-butylmethylphenoxide or hydride represented by the formula:

[30] Formula III-9 below: [ka] The method for preparing a compound represented by the following formula III-8: [ka] The method comprises the step of protecting the hydroxyl group attached to the carbon atom at position 2 of the D-mannopyranoside in a compound represented by formula III-1 with a benzyl group to produce a compound represented by formula III-9.

[31] Formula V-5 below: [ka] A method for producing a compound represented by the following formula V-4: [ka] with a strong base of the alkoxide type in the presence of methyl trifluoroacetate to produce a compound of formula V-5.

[32] The method according to

[31] , wherein the alkoxide-based strong base is selected from the group consisting of sodium salts, lithium salts, potassium salts of C1 to C5 alkoxides, and combinations thereof.

[33] Formula II-5 below: [ka] or a compound of formula II-8: [ka] A method for producing a compound represented by the formula: In fluorous alcohol and water, the following formula II-4: [ka] or a compound of formula II-7: [ka] A method comprising the step of reacting a compound represented by the formula: with λ3-iodane to produce a compound represented by formula II-5 or a compound represented by formula II-8.

[34] 33. The method of claim 33, wherein the λ3-iodane is selected from the group consisting of [bis(trifluoroacetoxy)iodo]benzene (PIFA), [hydroxy(tosyloxy)iodo]benzene (HTIB), (diacetoxyiodo)benzene (PIDA), [bis(trifluoroacetoxy)iodo]pentafluorobenzene, [hydroxy(methanesulfonyloxy)iodo]benzene, and combinations thereof.

[35] The method according to

[33] or

[34] , wherein the fluorous alcohol is selected from the group consisting of hexafluoro-2-propanol (HFIP), 2,2,2-trifluoroethanol (TFE), 2,2,3,3,4,4,5,5-octafluoro-1-pentanol, nonafluoro-tert-butyl alcohol, and combinations thereof.

[36] Formula II-6 below: [ka] or a compound of formula II-9: [ka] A method for preparing a compound represented by the following formula II-5: [ka] or a compound of formula II-8: [ka] in the presence of N-methylimidazole, with a compound of the formula: [ka] with 2,2,2-trifluoro-N-phenylacetimidoyl chloride (TFPC) of formula II-6 to produce a compound of formula II-9.

[37] Formula VI-5 below: [ka] A method for preparing a compound represented by the following formula VI-4: [ka] with a strong base of the alkoxide type in the presence of methyl trifluoroacetate to produce a compound of formula VI-5.

[38] The method according to

[37] , wherein the alkoxide-based strong base is selected from the group consisting of sodium salts, lithium salts, potassium salts of C1 to C5 alkoxides, and combinations thereof.

[39] Formula VII-3 below: [ka] The method for producing the compound represented by formula VII-3 comprises a step of contacting a solvent in which the compound represented by formula VII-3 is dissolved with silica gel to perform solid phase extraction of the compound represented by formula VII-3.

[40] Formula VIII-4 below: [ka] A method for preparing a compound represented by the following formula VIII-3: [ka] in the presence of N-methylimidazole, with a compound of the formula: [ka] with 2,2,2-trifluoro-N-phenylacetimidoyl chloride (TFPC) of formula VIII-4 to produce a compound of formula VIII-4.

[41] Formula IX-1 below: [ka] A method for producing a compound represented by the following formula VIII-5: [ka] and a compound represented by the following formula VII-3: [ka] by α-2,6-glycosidic bonding to form a compound represented by formula IX-1.

[42] Formula IX-1 below: [ka] The method for producing the compound represented by formula IX-1 comprises a step of contacting a solvent in which the compound represented by formula IX-1 is dissolved with silica gel to perform solid phase extraction of the compound represented by formula IX-1.

[43] The method according to

[42] , wherein the solvent in which the compound represented by formula IX-1 is dissolved is selected from the group consisting of toluene, dichloromethane, chloroform, and combinations thereof.

[44] Formula IX-3 below: [ka] The method for producing the compound represented by formula IX-3 comprises a step of contacting a solvent in which the compound represented by formula IX-3 is dissolved with silica gel to perform solid phase extraction of the compound represented by formula IX-3.

[45] The method according to

[44] , wherein the solvent in which the compound represented by formula IX-3 is dissolved is selected from the group consisting of toluene, dichloromethane, chloroform, and combinations thereof.

[46]

[23] to

[45] , comprising a compound of formula XI: [ka] A method for producing an oligosaccharide represented by the formula:

[47] Formula III-6 below: [ka] A compound represented by the formula:

[48] Formula III-10 below: [ka] A compound represented by the formula:

[49] Formula V-1 below: [ka] A compound represented by the formula:

[50] Formula V-2 below: [ka] A compound represented by the formula:

[51] Formula V-3 below: [ka] A compound represented by the formula:

[52] Formula II-6 below: [ka] A compound represented by the formula:

[53] Formula VI-A below: [ka] (Wherein R1 is [ka] A compound represented by the formula (I), wherein the compound is selected from the group consisting of:

[54] Formula VI-2 below: [ka] A compound represented by the formula:

[55] Formula VI-B below: [ka] (Wherein R2 is [ka] A compound represented by the formula (I), wherein the compound is selected from the group consisting of:

[56] Formula VIII-5 below: [ka] A compound represented by the formula:

[57] Formula X-1 below: [ka] A compound represented by the formula:

[58] Formula X-2 below: [ka] A compound represented by the formula:

[59] Formula X-3 below: [ka] A compound represented by the formula:

[60] Formula X-4 below: [ka] A compound represented by the formula:

[61] Formula X-5 below: [ka] A compound represented by the formula:

[62] Formula X-6 below: [ka] A compound represented by the formula:

[63] Formula X-7 below: [ka] A compound represented by the formula:

[64] A method for producing a glycochain remodeling antibody or an Fc region-containing molecule thereof, comprising: [1] to

[22] and

[46] , Further, a step of obtaining a sugar chain donor molecule containing N-acetylglucosamine (GlcNAc) whose reducing end is activated from the obtained oligosaccharide represented by formula XI; The method comprises the step of reacting the sugar chain donor molecule with an acceptor molecule which is an antibody having a core GlcNAc to which fucose may be attached as an N297-linked sugar chain, or an Fc region-containing molecule thereof.

[65] The production method described in

[64] , wherein the GlcNAc having an activated reducing end is an oxazolinated GlcNAc.

[66] The sugar chain donor molecule may be represented by the following formula XII, which may be chemically modified at the non-reducing end: [ka] The method according to

[64] or

[65] , wherein the compound is a compound represented by the formula (also referred to as "SG(10)-Ox").

[67] The method according to any one of

[64] to

[66] , wherein the sugar chain donor molecule is [N3-PEG(3)]2-SG(10)-Ox.

[68] The method according to

[67] , further comprising a step of reacting the azide group (N3-) with a molecule having an alkyne structure.

[69] The method of manufacturing according to

[68] , wherein the molecule having an alkyne structure is selected from chemotherapeutic agents, molecular targeted drugs, immune activators, toxins, antibacterial agents, antiviral agents, diagnostic agents, proteins, peptides, amino acids, nucleic acids, antigens, vitamins, and hormones.

[70] A method for producing an antibody-drug conjugate, comprising the method according to any one of

[64] to

[69] . [Effects of the Invention]

[0015] The present invention provides a novel purely chemical method for producing biantennary N-glycans having an α2,6-sialic acid structure at the non-reducing end, which is excellent in yield, selectivity, and efficiency. The present invention also provides novel methods for producing monosaccharides or oligosaccharides, intermediates, and novel methods for producing N-glycans that include these production methods, which are useful in the purely chemical production methods for N-glycans. Furthermore, the present invention provides novel methods for producing glycoproteins and the like (particularly, glycochain-remodeling antibodies or their FC region-containing molecules, or antibody-drug conjugates) that utilize the novel purely chemical production methods for N-glycans. [Brief explanation of the drawings]

[0016] [Figure 1] FIG. 1 is a simplified diagram showing a novel purely chemical method for producing biantennary N-glycans having an α2,6-sialic acid structure at the non-reducing end using a novel synthesis scheme provided by the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0017] Preferred embodiments for carrying out the present invention will be described below. Note that the embodiment described below shows one example of a typical embodiment of the present invention, and the scope of the present invention should not be construed as being narrow.

[0018] <1. A new purely chemical method for producing biantennary N-glycans with α2,6-sialic acid structures at the non-reducing end> In one aspect of the present invention, a novel purely chemical method for producing a biantennary N-glycan having an α2,6-sialic acid structure at the non-reducing end using a novel synthesis scheme is provided. In the present invention, the biantennary N-glycan having an α2,6-sialic acid structure at the non-reducing end is represented by the following formula XI: [ka] The oligosaccharide is represented by the formula:

[0019] The novel synthesis scheme of the present invention includes the following steps 1 to 5.

[0020] <Process 1> Step 1 involves reacting a compound of formula III-11 below: [ka] or a compound of formula III-13: [ka] Step 1 includes, as essential substeps, substeps 1-9 and 1-10 or substeps 1-13 and 1-14, which are related to the stereoinversion of glucose → mannose, as described below, but other steps can be carried out using or by applying conventional methods for the production of monosaccharides or oligosaccharides.

[0021] In one embodiment of the present invention, step 1 comprises the following substeps: [ka]

[0022] <Small process 1-1> Sub-step 1-1 is the reaction of the compound of formula I-1 below: [ka] The hydroxyl group bonded to the carbon atom at position 3 of the compound represented by the following formula I-2: [ka] The compound of formula I-1, which is the starting material for this step, can be prepared by known methods, or a commercially available product can be used. An example of a commercially available product of the compound of formula I-1 is 1,2:5,6-di-O-isopropylidene-α-D-glucofuranose manufactured by Sigma-Aldrich. This step can be carried out by utilizing or adapting known methods, but is preferably carried out by the method shown in Example 1, for example.

[0023] <Small process 1-2> Substep 1-2 is to convert the compound of formula I-2 to the compound of formula I-3 by acid hydrolysis of two isopropylidenes and pyranose ring formation: [ka] This step can be carried out by utilizing or applying a known method, but is preferably carried out by, for example, the method shown in Example 2.

[0024] <Small process 1-3> Substep 1-3 is to protect the hydroxyl group on the compound of formula I-3 with an acetyl group to give the compound of formula I-4: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 3.

[0025] <Small process 1-4> Sub-step 1-4 is to selectively remove only the acetyl group in the acetyloxy group bonded to carbon atom 1 of the compound represented by formula I-4 to obtain a compound represented by formula I-5: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 3.

[0026] <Small process 1-5> Substep 1-5 is the reaction of a compound of formula I-5 with trichloroacetonitrile to give a compound of formula I-6: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 4.

[0027] <Small process 1-6> Substep 1-6 is the reaction of a compound of formula I-6 with a compound of formula II-1: [ka] to give a compound of formula III-1: [ka] The compound represented by formula II-1 can be produced by known methods, or a commercially available product can be used. An example of a commercially available product of the compound represented by formula II-1 is 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-phthalimido-β-D-glucopyranoside manufactured by Tokyo Chemical Industry Co., Ltd. This step can be carried out by utilizing or adapting known methods, but is preferably carried out by, for example, the method shown in Example 5.

[0028] <Small process 1-7> Sub-step 1-7 is to remove the acetyl group from the compound of formula III-1 to obtain the compound of formula III-2: [ka] This step can be carried out by utilizing or applying a known method, but is preferably carried out by, for example, the method shown in Example 6 or 16.

[0029] In one embodiment of the present invention, substep 1-7 is a step in which a compound represented by formula III-1 is reacted with a strong alkoxide base in the presence of methyl trifluoroacetate to produce a compound represented by formula III-2. It has been reported that the acetyl group elimination reaction can be carried out using sodium methoxide in methanol (Org. Biomol. Chem., 2018, 16, 4720-4727). However, this can also result in an undesired side reaction, i.e., ring-opening of the phthalimide group. On the other hand, by using a method in which the compound is reacted with a strong alkoxide base in the presence of methyl trifluoroacetate, it is possible to eliminate the acetyl group while suppressing ring-opening of the phthalimide group.

[0030] The alkoxide-based strong base is not limited as long as the reaction proceeds, and examples thereof include sodium salts, lithium salts, and potassium salts of C1 to C5 alkoxides, and combinations thereof; preferred examples include lithium methoxide, sodium methoxide, potassium methoxide, lithium ethoxide, sodium ethoxide, potassium ethoxide, lithium isopropoxide, sodium isopropoxide, potassium isopropoxide, lithium tert-butoxide, sodium tert-butoxide, potassium tert-butoxide, lithium tert-pentoxide, sodium tert-pentoxide, and potassium tert-pentoxide; particularly preferred examples include sodium methoxide, lithium tert-butoxide, and potassium tert-butoxide.

[0031] The solvent used in this step is not limited as long as the reaction proceeds, and examples thereof include alcoholic solvents such as methanol, ethanol, propanol, and butanol, as well as mixed solvents of tetrahydrofuran, acetonitrile, cyclopentyl methyl ether, toluene, dimethylacetamide, and the like with alcoholic solvents, and preferred examples include methanol or mixed solvents of methanol and tetrahydrofuran.

[0032] The reaction temperature in this step is not limited as long as the reaction proceeds, but examples include -20°C to 80°C, preferably 0°C to 70°C, more preferably 20°C to 65°C, and particularly preferably 40°C to 60°C.

[0033] <Small process 1-8> Substep 1-8 is to selectively protect the hydroxyl groups attached to the 4- and 6-carbon atoms of the D-glucopyranoside in the compound of formula III-2 using benzaldehyde dimethyl acetal to obtain the compound of formula III-3: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by the method shown in Example 7 or 17, for example.

[0034] <Small process 1-9> Substep 1-9 is to react a compound of formula III-3 with a compound that provides a leaving group selected from the group consisting of a trifluoromethanesulfonyloxy group, a nonafluorobutanesulfonyloxy group, a 2-nitrobenzenesulfonyloxy group, and a 4-nitrobenzenesulfonyloxy group, to give a compound of formula III-4: [ka] (wherein X1 represents a substituent selected from the group consisting of a trifluoromethanesulfonyl group, a nonafluorobutanesulfonyl group, a 2-nitrobenzenesulfonyl group, and a 4-nitrobenzenesulfonyl group). This step can be carried out by utilizing or applying a known method for providing a leaving group, but is preferably carried out by, for example, the method shown in Example 8.

[0035] In this step, examples of the "compound that imparts a leaving group selected from the group consisting of a trifluoromethanesulfonyloxy group, a nonafluorobutanesulfonyloxy group, a 2-nitrobenzenesulfonyloxy group, and a 4-nitrobenzenesulfonyloxy group" include trifluoromethanesulfonic anhydride, nonafluoro-1-butanesulfonyl fluoride, bis(nonafluoro-1-butanesulfonic) anhydride, 2-nitrobenzenesulfonyl chloride, and 4-nitrobenzenesulfonyl chloride, and preferably trifluoromethanesulfonic anhydride.

[0036] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include ethyl acetate, toluene, dichloromethane, acetonitrile, cyclopentyl methyl ether, and tert-butyl methyl ether, and preferred examples include ethyl acetate, toluene, and dichloromethane.

[0037] The reaction temperature in this step is not limited as long as the reaction proceeds, but can be, for example, -40°C to 60°C, preferably -30°C to 40°C, and more preferably -20°C to 10°C.

[0038] This step is preferably carried out in the presence of a base. The base used in this step is not particularly limited as long as the reaction proceeds, but examples thereof include 1-methylimidazole, pyridine, 4-dimethylaminopyridine, picoline, lutidine, and collidine, and preferably 1-methylimidazole.

[0039] <Small process 1-10> Substep 1-10 is to react the compound of formula III-4 with cesium acetate or tetrabutylammonium acetate to give the compound of formula III-5: [ka] (wherein X2 is an acetyl group), or by reacting a compound of formula III-4 with tetrabutylammonium benzoate, a compound of formula III-5: [ka] (wherein X2 is a benzoyl group). Stereoinversion from glucose to mannose is a known conversion reaction, but no conversion has been reported in which the protecting group for the hydroxyl group attached to the carbon atom at the D-glucopyranoside position of a glucose-glucosamine disaccharide linked by a β-glycosidic bond is a 2-naphthylmethyl (Nap) group. By employing this method, stereoinversion from glucose to mannose can be achieved, and a mannose-glucosamine disaccharide skeleton linked by a β-glycosidic bond can be constructed with high yield and high selectivity.

[0040] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include dimethyl sulfoxide, dimethylformamide, dimethylacetamide, N-methylpyrrolidone, N,N-dimethylimidazolidinone, sulfolane, tetrahydrofuran, and acetonitrile, and preferred examples include dimethyl sulfoxide.

[0041] The reaction temperature in this step is not limited as long as the reaction proceeds, but examples include 20 to 80°C, preferably 23 to 70°C, more preferably 26 to 60°C, and particularly preferably 30 to 50°C.

[0042] This step can be preferably carried out by the method shown in Example 9, for example.

[0043] <Small process 1-11> Sub-step 1-11 is to remove the X2 group and open the phthalimide group in the compound of formula III-5 to give the compound of formula III-6: [ka] This step can be carried out by utilizing or applying a known hydrolysis method, but is preferably carried out by, for example, the method shown in Example 9.

[0044] The compound of formula III-6 produced in this step can be dissolved in a solvent and used directly in the next step, or it can be isolated and purified by recrystallization. The great advantage of crystallization is that it can be isolated and purified. Crystallization can almost completely remove impurities with similar structures that are difficult to remove by column purification. In this case, the compound of formula III-6 can be obtained with an HPLC purity of 99% or more.

[0045] The isolation and purification by recrystallization in this step is not limited as long as the reaction proceeds. For example, the method may involve completely removing the solvent from a dissolved state by drying under reduced pressure, or may involve using tetrahydrofuran as a good solvent and adding dropwise isopropanol as a poor solvent in the presence of a trace amount of water.

[0046] The recrystallization in this step can also be carried out using seed crystals of the compound represented by formula III-6. When seed crystals are used, for example, crystallization can be carried out by using tetrahydrofuran as a good solvent and adding a portion of isopropanol as a poor solvent dropwise in the presence of a trace amount of water, adding the seed crystals, confirming the precipitation of crystals, and then adding the remaining isopropanol dropwise.

[0047] <Small process 1-12> Substep 1-12 is to close the ring-opened phthalimide group in the compound of formula III-6 by dehydration condensation to obtain a compound of formula III-8: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by the method shown in Example 10 or 20, for example.

[0048] <Small process 1-15> Sub-step 1-15 is to protect the hydroxyl group attached to the carbon atom at the 2-position of D-mannopyranoside in the compound of formula III-8 with a benzyl group to obtain the compound of formula III-9: [ka] This step can be carried out by utilizing or applying a known method, but is preferably carried out by the method shown in Example 11 or 21, for example.

[0049] In one embodiment of the present invention, substep 1-15 comprises protecting the hydroxyl group attached to the carbon atom at position 2 of the D-mannopyranoside in the compound of formula III-8 with a benzyl group in the presence of lithium tert-butoxide or lithium tert-amoxide to produce a compound of formula III-9. By carrying out substep 1-15 in the presence of lithium tert-butoxide or lithium tert-amoxide, ring-opening of the phthalimide can be suppressed. Furthermore, compared to the general conditions using sodium hydride, this method is safer and easier to scale up.

[0050] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include dimethylacetamide, dimethylformamide, N-methylpyrrolidone, and N,N-dimethylimidazolidinone, and preferably dimethylacetamide.

[0051] The reaction temperature in this step is not limited as long as the reaction proceeds, but may be, for example, -20 to 100°C, preferably -15 to 70°C, and particularly preferably -10 to 50°C.

[0052] <Small process 1-16> Substep 1-16 is to selectively deprotect only the hydroxyl group bonded to the carbon atom at position 6 of the D-mannopyranoside in the compound of formula III-9 to obtain the compound of formula III-10: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 12 or 22.

[0053] In this step, the compound represented by formula III-10 produced in the solvent may be used as is in the next step, or it may be isolated and purified by column or recrystallization.

[0054] <Small process 1-17> Substep 1-17 is to remove the NAP group from the compound of formula III-10 to obtain the compound of formula III-11: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 13.

[0055] <Small process 1-18> Substep 1-18 is to remove the phthaloyl protecting group from the compound of formula III-11 to obtain the compound of formula III-12: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 14.

[0056] <Small process 1-19> Substep 1-19 is to protect the amino group in the compound of formula III-12 with 2,2,2-trichloroethoxycarbonyl group to give the compound of formula III-13: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 15.

[0057] In one embodiment of the present invention, step 1 involves the stereoinversion of glucose → mannose by S N Instead of sub-steps 1-9 to 1-12, which include steps for carrying out the reaction using two reactions, the following sub-steps 1-13 and 1-14, which are carried out using an oxidation-reduction reaction, are included.

[0058] [ka]

[0059] <Small process 1-13> Substep 1-13 is to oxidize the 2-position of D-glucopyranoside in the compound of formula III-3 to obtain the compound of formula III-7: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 18.

[0060] <Small process 1-14> Substep 1-14 is to reduce the oxo group attached to the carbon atom at the 2-position of 2-keto-D-glucopyranoside in the compound of formula III-7 to obtain a compound of formula III-8: [ka] This step can be carried out by utilizing or applying a known method, but is preferably carried out by, for example, the method shown in Example 19.

[0061] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include diethyl ether, cyclopentyl methyl ether, tert-butyl methyl ether, diisopropyl ether, dipropyl ether, dibutyl ether, and 1,4-dioxane, and preferably tetrahydrofuran.

[0062] The reaction temperature in this step is not limited as long as the reaction proceeds, and can be, for example, −80° C. to 20° C. As described below, the optimal reaction temperature varies depending on the reducing agent used.

[0063] In one embodiment of the present invention, the oxo group bonded to the carbon atom at the 2-position of 2-keto-D-glucopyranoside in the compound represented by formula III-7 can be substituted with L-selectride, LS-selectride, lithium diisobutyl-tert-butoxyaluminum hydride (LDBBA), or a compound represented by the following formula A: [ka] wherein R3 has the following formula: [ka] The reduction is carried out in the presence of a reducing agent selected from the group consisting of compounds represented by the formula (I) and combinations thereof, where R is a di-tert-butylmethylphenoxide or hydride represented by the formula (I), where at least two R are di-tert-butylmethylphenoxides. In this reduction step, for example, when NaBH is used, the stereoselectivity is low (approximately 7:3), making it difficult to efficiently achieve the desired stereoinversion from Gln to Man (Org. Biomol. Chem., 2018, 16, 4720-4727). On the other hand, when the reducing agents listed above are used, the selectivity of the stereoinversion from Gln to Man is significantly improved (93.6:6.4 to 98.1:1.9) compared to when NaBH is used.

[0064] Of the compounds represented by formula A, a compound in which three R3 are di-tert-butylmethylphenoxide can be obtained, for example, by adding dibutylhydroxytoluene (885.41 mg, 4.02 mmol) to a tetrahydrofuran suspension (2 mL) of lithium aluminum hydride (50.0 mg, 1.32 mmol) at 0° C., followed by stirring at 25° C. Of the compounds represented by formula A, a compound in which two R3 are di-tert-butylmethylphenoxide can be obtained in a similar manner by using 2 molar equivalents of dibutylhydroxytoluene for 1 molar equivalent of lithium aluminum hydride.

[0065] As described above, the reaction temperature in this step is not limited as long as the reaction proceeds, but when L-selectride, LS-selectride, or LDBBA is used as the reducing agent, the reaction temperature is preferably −80° C. to −20° C., more preferably −80° C. to −30° C., even more preferably −80° C. to −40° C., and particularly preferably −80° C. to −50° C. When a compound represented by formula A is used as the reducing agent, the reaction temperature is preferably −20° C. to 20° C., more preferably −15° C. to 15° C., and particularly preferably −10° C. to 10° C. Therefore, the compound represented by formula A is particularly preferred as the reducing agent used in this step, in that the reaction proceeds at a temperature that is easier to handle.

[0066] <Process 2> Step 2 is the reaction of the compound of formula V-3 below: [ka] or a compound represented by the following formula V-5: [ka] Step 2 includes substeps 2-3 and 2-6, which are described below, as essential substeps for regioselectively linking two monosaccharide molecules to one disaccharide molecule, but other steps can be carried out using or by applying conventional methods for producing monosaccharides or oligosaccharides.

[0067] In one embodiment of the present invention, step 2 comprises the following substeps: [ka] or [ka]

[0068] <Small process 2-1> Sub-step 2-1 is the reaction of the compound of formula IV-1: [ka] The acetyl group of the monoacetyl compound produced by hydrolysis of the orthoester moiety of the compound represented by the following formula IV-2: [ka] The compound of formula IV-1, which is the starting material of this step, can be prepared by known methods, or a commercially available product can be used. An example of a commercially available product of the compound of formula VI-1 is 3,4,6-Tri-O-benzyl-beta-D-mannopyranose-1,2-(methyl orthoacetate) manufactured by Combi-blocks. This step can be carried out by utilizing or adapting known methods, but is preferably carried out by the method shown in Example 23 or 26, for example.

[0069] <Small process 2-2> Substep 2-2 is the reaction of a compound of formula IV-2 with trichloroacetonitrile to give a compound of formula IV-3: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by the method shown in Example 23 or 27, for example.

[0070] <Small process 2-3> Substep 2-3 is to react a compound represented by formula III-13 with a compound represented by formula IV-3 to obtain a compound represented by formula V-1: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 24.

[0071] This step is preferably carried out in the presence of a Lewis acid. The Lewis acid used in this step is not limited as long as the reaction proceeds, but examples thereof include trimethylsilyl trifluoromethanesulfonate, triisopropylsilyl trifluoromethanesulfonate, tert-butyldimethylsilyl trifluoromethanesulfonate, and boron trifluoride diethyl ether complex, and preferably includes trimethylsilyl trifluoromethanesulfonate.

[0072] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include dichloromethane, toluene, trifluoromethylbenzene, and chlorobenzene, and preferred examples include dichloromethane and toluene.

[0073] The reaction temperature in this step is not limited as long as the reaction proceeds, but examples thereof include -60°C to 0°C, preferably -40°C to 0°C, more preferably -30°C to 0°C, and particularly preferably -20°C to 0°C.

[0074] In this step, it is preferable to add 2 to 4 equivalents of the compound represented by formula IV-3 to 1 equivalent of the compound represented by formula III-13, and it is more preferable to add 2.5 to 3 equivalents of the compound represented by formula IV-3 to 1 equivalent of the compound represented by formula III-13.

[0075] <Small process 2-4> Sub-step 2-4 is to obtain a compound of formula V-2 by removing the acetyl group and the 2,2,2-trichloroethoxycarbonyl group from the compound of formula V-1: [ka] This step can be carried out by utilizing or applying a known method, but is preferably carried out by, for example, the method shown in Example 25.

[0076] <Small process 2-5> Sub-step 2-5 is to protect the amino group in the compound of formula V-2 with a 2,2,2-trichloroethoxycarbonyl group to give the compound of formula V-3: [ka] This step can be carried out by utilizing or applying a known method, but is preferably carried out by, for example, the method shown in Example 25.

[0077] <Small process 2-6> Substep 2-6 is to react a compound of formula III-11 with a compound of formula IV-3 to obtain a compound of formula V-4: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 28.

[0078] This step is preferably carried out in the presence of a Lewis acid. The Lewis acid used in this step is not limited as long as the reaction proceeds, but examples thereof include trimethylsilyl trifluoromethanesulfonate, triisopropylsilyl trifluoromethanesulfonate, tert-butyldimethylsilyl trifluoromethanesulfonate, and boron trifluoride diethyl ether complex, and preferably includes trimethylsilyl trifluoromethanesulfonate.

[0079] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include dichloromethane, toluene, trifluoromethylbenzene, and chlorobenzene, and preferred examples include dichloromethane and toluene.

[0080] The reaction temperature in this step is not limited as long as the reaction proceeds, but examples thereof include -60°C to 0°C, preferably -40°C to 0°C, more preferably -30°C to 0°C, and particularly preferably -20°C to 0°C.

[0081] In this step, it is preferable to add 2 to 4 equivalents of the compound represented by formula IV-3 to 1 equivalent of the compound represented by formula III-11, and it is more preferable to add 2.5 to 3 equivalents of the compound represented by formula IV-3 to 1 equivalent of the compound represented by formula III-11.

[0082] <Small process 2-7> Sub-step 2-7 is to remove the acetyl group from the compound of formula V-4 to obtain the compound of formula V-5: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 29.

[0083] In one embodiment of the present invention, substep 2-7 comprises reacting a compound of formula V-4 with a strong alkoxide base in the presence of methyl trifluoroacetate to produce a compound of formula V-5. It is generally known that deprotection of the acetyl group is carried out using sodium methoxide in methanol, but this can also result in an undesired side reaction, i.e., ring-opening of the phthalimide group. On the other hand, by using a method of reacting with a strong alkoxide base in the presence of methyl trifluoroacetate, it is possible to remove the acetyl group while suppressing ring-opening of the phthalimide group.

[0084] The alkoxide-based strong base is not limited as long as the reaction proceeds, and examples thereof include sodium salts, lithium salts, and potassium salts of C1 to C5 alkoxides, and combinations thereof; preferred examples include lithium methoxide, sodium methoxide, potassium methoxide, lithium ethoxide, sodium ethoxide, potassium ethoxide, lithium isopropoxide, sodium isopropoxide, potassium isopropoxide, lithium tert-butoxide, sodium tert-butoxide, potassium tert-butoxide, lithium tert-pentoxide, sodium tert-pentoxide, and potassium tert-pentoxide; particularly preferred examples include sodium methoxide, lithium tert-butoxide, and potassium tert-butoxide.

[0085] The solvent used in this step is not limited as long as the reaction proceeds, and examples thereof include alcoholic solvents such as methanol, ethanol, propanol, and butanol, as well as mixed solvents of tetrahydrofuran, acetonitrile, cyclopentyl methyl ether, toluene, dimethylacetamide, and the like with alcoholic solvents, and preferred examples include methanol or mixed solvents of methanol and tetrahydrofuran.

[0086] The reaction temperature in this step is not limited as long as the reaction proceeds, but examples include -20°C to 80°C, preferably 0°C to 70°C, more preferably 15°C to 65°C, and particularly preferably 30°C to 60°C.

[0087] <Process 3> Step 3 is the reaction of the compound of formula VI-3 below: [ka] or a compound of formula VI-6: [ka] Step 3 includes substeps 3-6 and 3-12, which are described below, as essential substeps for regioselectively linking two monosaccharide molecules to one tetrasaccharide molecule, but other steps can be carried out using or by applying conventional methods for producing monosaccharides or oligosaccharides.

[0088] In one embodiment of the present invention, step 3 comprises the following substeps: [ka] Or, [ka]

[0089] <Small process 3-1> Sub-step 3-1 is the reaction of the compound of formula II-1 below: [ka] The phthaloyl protecting group is removed from the compound represented by the following formula II-2: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 30.

[0090] <Small process 3-2> Sub-step 3-2 is to protect the amino group in the compound of formula II-2 with a 2,2,2-trichloroethoxycarbonyl group to give the compound of formula II-3: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 31.

[0091] <Small process 3-3> Sub-step 3-3 is to protect the hydroxyl group on the compound of formula II-3 with an acetyl group to give the compound of formula II-4: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 32.

[0092] <Small process 3-4> Sub-step 3-4 is to remove the 4-methoxyphenyl group from the compound of formula II-4 to obtain the compound of formula II-5: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 33.

[0093] In one embodiment of the present invention, this step is a step of producing a compound represented by formula II-5 by reacting a compound represented by formula II-4 with λ3-iodane in a fluorous alcohol and water.

[0094] In the present invention, "λ3-iodane" refers to a trivalent hypervalent iodine compound. The use of λ3-iodane not only increases the yield of the compound represented by Formula II-5 compared to conventional deprotection methods, but also allows the reaction to be carried out under milder reaction conditions. Furthermore, because the reaction proceeds with a slight excess of λ3-iodane, purification of the deprotected product is easier and operational safety is higher than in conventional deprotection methods that use, for example, an excess of ammonium cerium (IV) nitrate.

[0095] In one embodiment, the λ-iodane has the formula R 4 -I(OR 5 ) 2 (wherein R 4 is an unsubstituted or substituted phenyl group, and R 5 is selected from the group consisting of H, acetyl, trifluoroacetyl, tosyl, methanesulfonyl, and combinations thereof. As defined in the formula above, R 4may be a "substituted phenyl group," and examples of the substituent include a linear or branched saturated or unsaturated hydrocarbon group, an oxygen-containing group (alkoxy, ester, etc.), a nitrogen-containing group (cyano, azide, etc.), and a halogen (e.g., fluorine, chlorine, bromine, iodine), with a hydrocarbon group, an oxygen-containing substituent, or a halogen being more preferred. When these substituents contain carbon, for example, those having 1 to 5 carbon atoms or those having 1 to 3 carbon atoms can be suitably used. Specific examples of λ3-iodane include, but are not limited to, [bis(trifluoroacetoxy)iodo]benzene (PIFA), [hydroxy(tosyloxy)iodo]benzene (HTIB), (diacetoxyiodo)benzene (PIDA), [bis(trifluoroacetoxy)iodo]pentafluorobenzene, and [hydroxy(methanesulfonyloxy)iodo]benzene.

[0096] The amount of λ3-iodane can be appropriately set, for example, about 1 to 10 equivalents, about 1 to 7 equivalents, or about 1 to 5 equivalents relative to the compound represented by Formula II-4, and preferably about 1 to 3 equivalents. Throughout this specification, the term "about" indicates a range of ±10% of the stated value.

[0097] In the present invention, "fluorous alcohol" refers to a fluorine-containing alcohol compound in which all carbon atoms except the carbon bonded to the alcohol have fluorine. As long as fluorine substitution is permitted, it is preferable for the fluorous alcohol to have more fluorine atoms. Fluorous alcohols include, but are not limited to, fluorous aliphatic alcohols. The hydrocarbon moiety in the fluorous aliphatic alcohol may be saturated or unsaturated, linear or branched, or cyclic. Examples of fluorous aliphatic alcohols include fluorous C2-C8 aliphatic alcohols, preferably fluorous C2-C5 aliphatic alcohols, and more preferably fluorous C2-C3 aliphatic alcohols. Specific examples of fluorous alcohols include, but are not limited to, hexafluoro-2-propanol (HFIP), 2,2,2-trifluoroethanol (TFE), 2,2,3,3,4,4,5,5-octafluoro-1-pentanol, nonafluoro-tert-butyl alcohol, and combinations thereof.

[0098] Furthermore, preferred combinations of fucolic alcohol and λ3-iodane can provide the deprotected product in higher yields. Those skilled in the art can select appropriate combinations, but examples include, but are not limited to, PIFA preferably used in combination with hexafluoro-2-propanol (HFIP), 2,2,2-trifluoroethanol (TFE), nonafluoro-tert-butyl alcohol, HTIB preferably used in combination with HFIP, TFE, 2,2,3,3,4,4,5,5-octafluoro-1-pentanol, [bis(trifluoroacetoxy)iodo]pentafluorobenzene preferably used in combination with hexafluoro-2-propanol (HFIP), and [hydroxy(methanesulfonyloxy)iodo]benzene preferably used in combination with hexafluoro-2-propanol (HFIP).

[0099] The amount of fluorous alcohol can be appropriately set from the viewpoint of achieving a high yield of the product, and may be, for example, about 1.0 equivalent or more, about 1.5 equivalents or more, about 2.0 equivalents or more, or 2.5 equivalents or more in molar ratio relative to the compound represented by Formula II-4, and may be about 15 or less, about 10 or less, about 8 or less, or about 5 or less in volume ratio relative to the compound represented by Formula II-4.

[0100] This step is carried out in the presence of the fluorous alcohol and water. The amount of water can be appropriately determined from the viewpoint of achieving a high yield of the product, and may be, for example, about 1.0 equivalent or more, about 1.5 equivalents or more, about 2.0 equivalents or more, or about 2.5 equivalents or more in molar ratio relative to the compound of Formula II-4, and about 10 or less, about 8 or less, about 5 or less, or about 3 or less in volume ratio relative to the compound of Formula II-4.

[0101] In this step, a "solvent" may be further added to the fluorous alcohol and water. The solvent may be selected from the group consisting of, but not limited to, dichloromethane, toluene, (trifluoromethyl)benzene, and combinations thereof. The type of solvent used may be appropriately selected depending on the λ3-iodane used, etc., in order to achieve a high yield of the product. The amount of solvent may also be appropriately set in order to achieve a high yield of the product, and may be, for example, about 0.5 to 50, about 1 to 20, or about 2 to 10 by volume relative to the compound represented by Formula II-4.

[0102] In this step, an "additive" may be further added to the fluorous alcohol and water. The additive is preferably selected from the group consisting of sodium dihydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, and combinations thereof. Since the acidity may increase as the 4-methoxyphenylation reaction proceeds, adding an additive such as potassium dihydrogen phosphate can achieve a higher product yield, particularly when using λ3-iodane (HTIB, etc.), which by-produces a strongly acidic acid. The amount of the additive can also be appropriately selected to achieve a high product yield; for example, it can be about 0.5 to 8 equivalents, about 1 to 6 equivalents, or about 1.5 to 5 equivalents relative to the compound represented by Formula II-4.

[0103] When (diacetoxyiodo)benzene (PIDA) is used as the λ3-iodane, it is preferable to add trifluoroacetic acid (TFA) to obtain the deprotected product in higher yield.

[0104] In this step, in the presence of a fluorous alcohol and water, λ3-iodane acts as an oxidizing agent on the 4-methoxyphenyl (X group) in the compound of Formula II-4, resulting in the elimination of the OX group from the compound of Formula II-4. Water (HO) then adds to this oxidizing agent, facilitating the elimination of the 4-methoxyphenoxy (OX group) from the compound of Formula II-4. This action of λ3-iodane is generally achieved by stirring, refluxing, or otherwise treating a solution containing the compound of Formula II-4 and λ3-iodane in a fluorous alcohol and water. Therefore, this step can be easily carried out and scaled up. The product, the compound of Formula II-5, can be purified or isolated by any purification method known to those skilled in the art, such as crystallization or chromatography.

[0105] The temperature when reacting the compound represented by Formula II-4 with λ3-iodane is preferably from about −20° C. to the boiling point of the fluorous alcohol (e.g., about 58° C. for hexafluoro-2-propanol (HFIP) and about 78° C. for 2,2,2-trifluoroethanol (TFE)) or lower, and can be, for example, about −20 to 60° C., about 0 to 60° C., or about 10 to 30° C. Another advantage is that the demethoxyphenylation reaction proceeds even at room temperature (15 to 30° C.), eliminating the need for cooling or heating.

[0106] The reaction time can be appropriately set to obtain the product in high yield.

[0107] <Small process 3-5> Substep 3-5 is the reaction of a compound of formula II-5 with a compound of the following formula: [ka] by reacting with 2,2,2-trifluoro-N-phenylacetimidoyl chloride (TFPC) of the following formula II-6: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 34.

[0108] In one embodiment of the present invention, this step involves reacting a compound of formula II-5 with TFPC in the presence of N-methylimidazole to produce a compound of formula II-6. For example, under certain conditions, when potassium carbonate is used as a base in this step, the yield is approximately 36%, but when N-methylimidazole is used, the yield is significantly improved to approximately 76%. Since TFPC is an expensive reagent, improving the yield of this step is very beneficial for commercial production.

[0109] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include dichloromethane, toluene, ethyl acetate, acetonitrile, and tetrahydrofuran, and preferably dichloromethane.

[0110] The reaction temperature in this step is not limited as long as the reaction proceeds, but is preferably -20 to 40°C, more preferably -10 to 35°C, and particularly preferably 0 to 30°C.

[0111] This step is preferably carried out in the presence of a dehydrating agent. The dehydrating agent used in this step is not limited as long as the reaction proceeds, but examples thereof include molecular sieves, and preferably molecular sieves 4A powder with a powder particle size of 10 μm or less.

[0112] In this step, the compound of Formula II-6 produced in the solvent may be used directly in the next step, or it may be isolated and purified by column chromatography or recrystallization. Examples of column chromatography include isolation and purification using silica gel as the stationary phase and dichloromethane or a toluene-ethyl acetate mixed solvent system as the mobile phase. Examples of recrystallization include crystallization from a diisopropyl ether and heptane mixed solvent system. Recrystallization can be preferably performed using seed crystals of the compound of Formula II-6. When using seed crystals, the compound can be obtained by, for example, inoculating the seed crystals into a diisopropyl ether solution, confirming the precipitation of crystals, and then adding heptane dropwise.

[0113] <Small process 3-6> Substep 3-6 is to react a compound represented by formula V-3 with a compound represented by formula II-6 to obtain a compound represented by formula VI-1: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 35.

[0114] This step is preferably carried out in the presence of a Lewis acid. The Lewis acid used in this step is not limited as long as the reaction proceeds, but examples thereof include trimethylsilyl trifluoromethanesulfonate, triisopropylsilyl trifluoromethanesulfonate, tert-butyldimethylsilyl trifluoromethanesulfonate, and boron trifluoride diethyl ether complex, and preferred examples thereof include trimethylsilyl trifluoromethanesulfonate, triisopropylsilyl trifluoromethanesulfonate, and tert-butyldimethylsilyl trifluoromethanesulfonate.

[0115] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include dichloromethane, toluene, trifluoromethylbenzene, and chlorobenzene, and preferably dichloromethane.

[0116] The reaction temperature in this step is not limited as long as the reaction proceeds, but examples thereof include -78°C to 0°C, preferably -78 to -20°C, more preferably -78 to -30°C, and particularly preferably -78 to -40°C.

[0117] In this step, it is preferable to add 2 to 5 equivalents of the compound represented by formula II-6 to 1 equivalent of the compound represented by formula V-3, and it is more preferable to add 3 to 5 equivalents of the compound represented by formula II-6 to 1 equivalent of the compound represented by formula V-3.

[0118] <Small process 3-7> Sub-step 3-7 is to remove the acetyl group and the 2,2,2-trichloroethoxycarbonyl group from the compound of formula VI-1 to obtain a compound of formula VI-2: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 36.

[0119] <Small process 3-8> Sub-step 3-8 is to protect the amino group in the compound of formula VI-2 with a 2,2,2-trichloroethoxycarbonyl group to give the compound of formula VI-3: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 36.

[0120] <Small process 3-9> Substep 3-9 is the reaction of the compound of formula II-1 below: [ka] With an acetyl group, the hydroxyl group on the compound of formula II-7: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 37.

[0121] <Small process 3-10> Sub-step 3-10 is to remove the 4-methoxyphenyl group from the compound of formula II-7 to obtain the compound of formula II-8: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 38.

[0122] In one embodiment of the present invention, this step is a step of producing a compound represented by formula II-8 by reacting a compound represented by formula II-7 with λ3-iodane in fluorous alcohol and water. Details of this step are applicable mutatis mutandis to the description of substep 3-4 in this specification, where "a compound represented by formula II-4" and "a compound represented by formula II-5" are read as "a compound represented by formula II-7" and "a compound represented by formula II-8," respectively.

[0123] <Small process 3-11> Substep 3-11 is the reaction of a compound of formula II-8 with a compound of the following formula: [ka] By reacting with 2,2,2-trifluoro-N-phenylacetimidoyl chloride (TFPC) represented by the following formula II-9: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 39.

[0124] In one embodiment of the present invention, this step involves reacting a compound of formula II-8 with TFPC in the presence of N-methylimidazole to produce a compound of formula II-9. The use of N-methylimidazole as a base allows for a reduction in the amount of TFPC, while still providing the desired product in high yield. Because TFPC is an expensive reagent, improving the yield of this step is highly beneficial for commercial production.

[0125] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include dichloromethane, toluene, ethyl acetate, acetonitrile, and tetrahydrofuran, and preferably dichloromethane.

[0126] The reaction temperature in this step is not limited as long as the reaction proceeds, but is preferably -20 to 40°C, more preferably -10 to 35°C, and particularly preferably 0 to 30°C.

[0127] This step is preferably carried out in the presence of a dehydrating agent. The dehydrating agent used in this step is not limited as long as the reaction proceeds, but examples thereof include molecular sieves, and preferably molecular sieves 4A powder with a powder particle size of 10 μm or less.

[0128] <Small process 3-12> Substep 3-12 is to react a compound represented by formula V-5 with a compound represented by formula II-9 to obtain a compound represented by formula VI-4: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 40.

[0129] This step is preferably carried out in the presence of a Lewis acid. The Lewis acid used in this step is not limited as long as the reaction proceeds, but examples thereof include trimethylsilyl trifluoromethanesulfonate, triisopropylsilyl trifluoromethanesulfonate, tert-butyldimethylsilyl trifluoromethanesulfonate, and boron trifluoride diethyl ether complex, and preferred examples thereof include trimethylsilyl trifluoromethanesulfonate, triisopropylsilyl trifluoromethanesulfonate, and tert-butyldimethylsilyl trifluoromethanesulfonate.

[0130] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include dichloromethane, toluene, trifluoromethylbenzene, and chlorobenzene, and preferably dichloromethane.

[0131] The reaction temperature in this step is not limited as long as the reaction proceeds, but examples thereof include -78°C to 0°C, preferably -78 to -20°C, more preferably -78 to -30°C, and particularly preferably -78 to -40°C.

[0132] In this step, it is preferable to add 2 to 5 equivalents of the compound represented by formula II-9 to 1 equivalent of the compound represented by formula V-5, and it is more preferable to add 3 to 5 equivalents of the compound represented by formula II-9 to 1 equivalent of the compound represented by formula V-5.

[0133] <Small process 3-13> Sub-step 3-13 is to remove the acetyl group from the compound of formula VI-4 to obtain the compound of formula VI-5: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 41.

[0134] In one embodiment of the present invention, substep 3-13 is a step of producing a compound represented by formula VI-5 by reacting a compound represented by formula VI-4 with a strong alkoxide base in the presence of methyl trifluoroacetate. It is generally known that deprotection of the acetyl group is carried out using sodium methoxide in methanol, but this can also result in an undesired side reaction, i.e., ring-opening of the phthalimide group. On the other hand, by using a method of reacting with a strong alkoxide base in the presence of methyl trifluoroacetate, it is possible to remove the acetyl group while suppressing ring-opening of the phthalimide group.

[0135] The alkoxide-based strong base is not limited as long as the reaction proceeds, and examples thereof include sodium salts, lithium salts, and potassium salts of C1 to C5 alkoxides, and combinations thereof; preferred examples include lithium methoxide, sodium methoxide, potassium methoxide, lithium ethoxide, sodium ethoxide, potassium ethoxide, lithium isopropoxide, sodium isopropoxide, potassium isopropoxide, lithium tert-butoxide, sodium tert-butoxide, potassium tert-butoxide, lithium tert-pentoxide, sodium tert-pentoxide, and potassium tert-pentoxide; particularly preferred examples include sodium methoxide, lithium tert-butoxide, and potassium tert-butoxide.

[0136] The solvent used in this step is not limited as long as the reaction proceeds, and examples thereof include alcoholic solvents such as methanol, ethanol, propanol, and butanol, as well as mixed solvents of tetrahydrofuran, acetonitrile, cyclopentyl methyl ether, toluene, dimethylacetamide, and the like with alcoholic solvents, and preferred examples include methanol or mixed solvents of methanol and tetrahydrofuran.

[0137] The reaction temperature in this step is not limited as long as the reaction proceeds, but examples include -20°C to 80°C, preferably 0°C to 70°C, more preferably 15°C to 65°C, and particularly preferably 30°C to 60°C.

[0138] <Small process 3-14> Sub-step 3-14 is to remove the phthaloyl group from the compound of formula VI-5 to obtain the compound of formula VI-2: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 42.

[0139] <Small process 3-15> Sub-step 3-15 is to protect the amino group in the compound of formula VI-2 with a 2,2,2-trichloroethoxycarbonyl group to give the compound of formula VI-3: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 43.

[0140] <Small process 3-16> Sub-step 3-16 is to protect the amino group in the compound of formula VI-2 with an acetyl group to obtain the compound of formula VI-6: [ka] This step can be carried out by utilizing or applying a known method, but is preferably carried out by, for example, the method shown in Example 44.

[0141] <Step 4> Step 4 is the reaction of a compound of formula IX-5: [ka] Step 4 includes substeps 4-7, which are described below, as essential substeps in which two monosaccharide molecules are linked via an α-2,6-glycosidic bond to synthesize a disaccharide block. However, other steps can be carried out using or by applying conventional methods for the production of monosaccharides or oligosaccharides.

[0142] In one embodiment of the present invention, step 4 comprises the following substeps: [ka]

[0143] <Small process 4-1> Sub-step 4-1 is the reaction of the compound of formula VII-1: [ka] with a benzoyl group to obtain the compound of formula VII-2: [ka] The compound represented by formula VII-1, which is the starting material of this step, is a compound identified by CAS No. 100759-10-2 and can be produced by a known method. This step can be carried out by utilizing or applying a known method, but is preferably carried out by, for example, the method shown in Example 45.

[0144] <Small process 4-2> Substep 4-2 is to remove the benzylidene protecting group from the compound of formula VII-2 to obtain the compound of formula VII-3: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 46.

[0145] In one embodiment of the present invention, this step includes a step of contacting the solvent containing the compound of formula VII-3 with silica gel to extract the compound of formula VII-3. Since the unreacted compound of formula VII-2 and the released benzaldehyde are not adsorbed to silica gel, this step allows for efficient purification of the compound of formula VII-3.

[0146] Examples of solvents for dissolving the compound represented by formula VII-3 include toluene, heptane, dichloromethane, chloroform, and combinations thereof. Preferred examples include toluene, dichloromethane, chloroform, and combinations thereof. Particularly preferred examples include, but are not limited to, toluene.

[0147] The amount of silica gel used in this step can be, for example, 2 to 5 times the amount of the raw materials, preferably 2 to 4 times the amount of the raw materials, and more preferably about 3 times the amount of the raw materials.

[0148] In this step, the solvent for eluting the compound of formula VII-3 adsorbed on the silica gel is not particularly limited as long as it does not dissolve the silica gel and can elute the target compound. Examples of the solvent include cyclopentyl methyl ether, ethyl acetate, and tert-butyl methyl ether.

[0149] <Small process 4-3> Sub-step 4-3 is the reaction of the compound of formula VIII-1: [ka] and then adding water to obtain a compound represented by the following formula VIII-2: [ka] The compound represented by formula VIII-1, which is the starting material of this step, can be prepared by known methods, or a commercially available product can be used. An example of a commercially available product of the compound represented by formula VIII-1 is N-acetylneuraminic acid manufactured by Tokyo Chemical Industry Co., Ltd. This step can be carried out by utilizing or adapting known methods, but is preferably carried out by the method shown in Example 47, for example.

[0150] <Small process 4-4> Sub-step 4-4 is to selectively protect hydroxyl groups other than the hydroxyl group bonded to carbon 1 in the compound represented by formula VIII-2 with acetyl groups to obtain a compound represented by formula VIII-3: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 48.

[0151] <Small process 4-5> Sub-step 4-5 is to react the compound of formula VIII-3 with 2,2,2-trifluoro-N-phenylacetimidoyl chloride (TFPC) to give the compound of formula VIII-4: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 49.

[0152] In one embodiment of the present invention, this step involves reacting a compound of formula VIII-3 with TFPC in the presence of N-methylimidazole to produce a compound of formula VIII-4. Compared to the use of potassium carbonate as the base in this step, the use of N-methylimidazole allows for a reduction in the amount of TFPC required, and still allows for a high yield of the target compound. Because TFPC is an expensive reagent, improving the yield of this step is highly beneficial for commercial production.

[0153] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include dichloromethane, toluene, ethyl acetate, acetonitrile, and tetrahydrofuran, and preferably dichloromethane.

[0154] The reaction temperature in this step is not limited as long as the reaction proceeds, but is preferably 20 to 40°C, more preferably 10 to 35°C, and particularly preferably 0 to 30°C.

[0155] This step is preferably carried out in the presence of a dehydrating agent. The dehydrating agent used in this step is not limited as long as the reaction proceeds, but examples thereof include molecular sieves, and preferably molecular sieves 4A powder with a powder particle size of 10 μm or less.

[0156] <Small process 4-6> Sub-step 4-6 is to protect the nitrogen atom in the acetamide group of the compound of formula VIII-4 with a tert-butoxycarbonyl group to give the compound of formula VIII-5: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 50.

[0157] In this step, the compound represented by formula VIII-5 produced in the solvent may be used directly in the next step, or it may be isolated and purified by recrystallization. The major advantage of the compound represented by formula VIII-5 is that it can be isolated and purified by crystallization. Crystallization allows the compound represented by formula VIII-5 to be obtained with an HPLC purity of 99% or more, and since it contains no impurities, it is possible to stably carry out the glycosylation reaction in the next step. Isolation and purification by recrystallization can be performed, for example, by adding heptane to a solution of cyclopentyl methyl ether to crystallize it.

[0158] <Small process 4-7> Sub-step 4-7 is to form a compound represented by formula VIII-5 and a compound represented by formula VII-3 via α-2,6-glycosidic bond to give a compound represented by formula IX-1: [ka] This process produces a compound represented by the formula: (Illegible text - likely OCR error). It is difficult to selectively conjugate an N-acetylneuraminic acid derivative and a galactose derivative via an α-2,6-glycosidic bond. For example, a method for synthesizing a disaccharide by reacting a compound represented by formula VIII-4 with a compound represented by formula VII-3 has been reported (J. Org. Chem., 2016, 81, 10600-10616). However, the reaction was difficult to reproduce, and the desired yield and selectivity could not be achieved. Furthermore, this reaction had issues such as a decrease in selectivity with increasing scale, a narrow reaction temperature tolerance, and a significant effect of reaction heat. Because the compound represented by formula VIII-4, one of the raw materials for this reaction, is very expensive, the low reproducibility, yield, and selectivity of this reaction pose a major problem, especially for commercial production, which requires scale-up. On the other hand, when the compound represented by formula VIII-5, which has a tert-butoxycarbonyl group added thereto, is used as the starting compound instead of the compound represented by formula VIII-4, high selectivity for α-2,6-glycosidic bonds (α:β=93:7) can be achieved with good reproducibility and improved yield. Furthermore, the temperature tolerance is broadened, and high reproducibility, yield, and selectivity can be achieved even when scaled up. This is extremely beneficial for commercial production.

[0159] This step is preferably carried out in the presence of a Lewis acid. The Lewis acid used in this step is not limited as long as the reaction proceeds, but examples thereof include trimethylsilyl trifluoromethanesulfonate, triisopropylsilyl trifluoromethanesulfonate, and tert-butyldimethylsilyl trifluoromethanesulfonate, and preferably trimethylsilyl trifluoromethanesulfonate.

[0160] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include diisopropyl ether, tert-butyl methyl ether, diethyl ether, dibutyl ether, dipropyl ether, 1,4-dioxane, dichloromethane, 1,2-dichloroethane, toluene, chlorobenzene, trifluoromethylbenzene, propionitrile, and acetonitrile, and preferred examples include cyclopentyl methyl ether.

[0161] The reaction temperature in this step is not limited as long as the reaction proceeds, but examples thereof include -78°C to 0°C, preferably -78 to -20°C, more preferably -78 to -30°C, and particularly preferably -78 to -40°C.

[0162] In this step, it is preferable to add 1 to 3 equivalents of the compound represented by formula VII-3 to 1 equivalent of the compound represented by formula VIII-5, and it is more preferable to add 1.4 to 2 equivalents of the compound represented by formula VII-3 to 1 equivalent of the compound represented by formula VIII-5.

[0163] This step can be carried out, for example, by adding a mixed solution of the compound represented by Formula VIII-5 and the compound represented by Formula VII-3 (preferably a cyclopentyl methyl ether mixed solution) dropwise to a solution containing a Lewis acid (preferably a cyclopentyl methyl ether solution) for a long period of time, or by adding a solution of the compound represented by Formula VIII-5 (preferably a cyclopentyl methyl ether solution) dropwise to a solution containing a Lewis acid and the compound represented by Formula VII-3 (preferably a cyclopentyl methyl ether solution) for a long period of time. The addition time is not limited as long as the reaction proceeds, but is, for example, 30 minutes to 5 hours, preferably 1 to 4 hours, more preferably 2 to 3.5 hours, and particularly preferably about 3 hours.

[0164] In one embodiment of the present invention, this step includes a step of solid-phase extraction of the compound of formula IX-1 by contacting a solvent containing the compound of formula IX-1 with silica gel. Since N-phenyltrifluoroacetamide, a by-product of the glycosylation reaction, and other trace impurities in the toluene solvent that are not adsorbed by silica gel are not adsorbed by silica gel, the compound of formula IX-1 can be efficiently purified by this step.

[0165] Examples of solvents for dissolving the compound represented by formula IX-1 include toluene, heptane, dichloromethane, chloroform, and combinations thereof. Preferred examples include toluene, dichloromethane, chloroform, and combinations thereof. Particularly preferred examples include, but are not limited to, toluene.

[0166] The amount of silica gel used in this step can be, for example, 2 to 5 times the amount of the raw materials, preferably 2 to 4 times the amount of the raw materials, and more preferably about 3.5 times the amount of the raw materials.

[0167] In this step, the solvent for eluting the compound represented by formula IX-1 adsorbed on the silica gel is not particularly limited as long as it does not dissolve the silica gel and can elute the target compound. Examples of the solvent include ethyl acetate, cyclopentyl methyl ether, and tert-butyl methyl ether, and preferably ethyl acetate.

[0168] This step can be carried out, for example, by the method shown in Example 51.

[0169] <Small process 4-8> Sub-step 4-8 is to remove the tert-butoxycarbonyl group from the compound of formula IX-1 to obtain the compound of formula IX-2: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 52.

[0170] <Small process 4-9> Sub-step 4-9 is to further protect the hydroxyl group of the compound of formula IX-2 and the nitrogen atom in the acetamide group with an acetyl group to give the compound of formula IX-3: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 53.

[0171] In one embodiment of the present invention, this step involves contacting a solvent containing the compound of formula IX-3 with silica gel to perform solid-phase extraction of the compound of formula IX-3. By-products such as the diacetyl form of the compound of formula VII-3, which is produced by acetylation of the excess compound of formula VII-3 used in the upstream glycosylation reaction, are not adsorbed by silica gel, and thus the compound of formula IX-3 can be efficiently purified by this step.

[0172] Examples of solvents for dissolving the compound represented by formula IX-3 include toluene, heptane, dichloromethane, chloroform, and combinations thereof. Preferred examples include toluene, dichloromethane, chloroform, and combinations thereof. Particularly preferred examples include, but are not limited to, toluene.

[0173] The amount of silica gel used in this step can be, for example, 2 to 5 times the amount of the raw materials, preferably 2 to 4 times the amount of the raw materials, and more preferably about 3.5 times the amount of the raw materials.

[0174] In this step, the solvent for eluting the compound represented by formula IX-3 adsorbed on the silica gel is not particularly limited as long as it does not dissolve the silica gel and can elute the target compound. Examples of the solvent include ethyl acetate, cyclopentyl methyl ether, and tert-butyl methyl ether, and preferably ethyl acetate.

[0175] <Small process 4-10> Sub-step 4-10 is to remove the allyl group attached to the carbon atom at position 1 of the D-galactopyranoside in the compound of formula IX-3 to obtain the compound of formula IX-4: [ka] This step can be carried out by utilizing or applying a known method, but is preferably carried out by, for example, the method shown in Example 54-1 and / or Example 54-2.

[0176] <Small process 4-11> Substep 4-11 is the reaction of a compound of formula IX-4 with 2,2,2-trifluoro-N-phenylacetimidoyl chloride (TFPC) to give the compound of formula IX-5: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 55.

[0177] <Process 5> Step 5 is to react a compound of formula XI below: [ka] Step 5 includes substeps 5-1 and 5-9, which are described below, as essential substeps for linking two monosaccharide molecules to one hexasaccharide molecule, but other steps can be carried out by using or applying conventional methods for producing monosaccharides or oligosaccharides.

[0178] In one embodiment of the present invention, step 5 comprises the following substeps:

[0179] [ka] [ka]

[0180] <Small process 5-1> Substep 5-1 is to react a compound represented by formula VI-3 with a compound represented by formula IX-5 to obtain a compound represented by formula X-1: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 56.

[0181] This step is preferably carried out in the presence of a Lewis acid. The Lewis acid used in this step is not limited as long as the reaction proceeds, but examples thereof include trimethylsilyl trifluoromethanesulfonate, triisopropylsilyl trifluoromethanesulfonate, and tert-butyldimethylsilyl trifluoromethanesulfonate, and preferred examples include tert-butyldimethylsilyl trifluoromethanesulfonate.

[0182] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include dichloromethane, toluene, trifluoromethylbenzene, and chlorobenzene.

[0183] The reaction temperature in this step is not limited as long as the reaction proceeds, but examples include -40°C to 20°C, preferably -30 to 10°C, more preferably -20 to 5°C, and particularly preferably -10 to 0°C.

[0184] In this step, it is preferable to add 2 to 5 equivalents of the compound represented by formula IX-5 to 1 equivalent of the compound represented by formula VI-3, and it is more preferable to add 3 to 4 equivalents of the compound represented by formula IX-5 to 1 equivalent of the compound represented by formula VI-3.

[0185] <Small process 5-2> Sub-step 5-2 is to remove the 2,2,2-trichloroethoxycarbonyl group from a compound of formula X-1 to obtain a compound of formula X-2: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 57.

[0186] <Small process 5-3> Sub-step 5-3 is to protect the amino group in the compound of formula X-2 with an acetyl group to obtain the compound of formula X-3: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 58.

[0187] <Small process 5-4> Sub-step 5-4 is to remove only one acetyl group from the diacetylamino group on D-galacto-non-2-ulopyranoside in the compound of formula X-3 to obtain the compound of formula X-4: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 59.

[0188] <Small process 5-5> Sub-step 5-5 is to remove the benzyl group from the compound of formula X-4 to obtain the compound of formula X-5: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 60.

[0189] <Small process 5-6> Sub-step 5-6 is to protect the hydroxyl group of the compound of formula X-5 with an acetyl group to obtain the compound of formula X-6: [ka] This step can be carried out by utilizing or applying a known method, but is preferably carried out by, for example, the method shown in Example 61.

[0190] <Small process 5-7> Sub-step 5-7 is to remove the 4-methoxyphenyl group from the compound of formula X-6 to obtain the compound of formula X-7: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 62.

[0191] <Small process 5-8> In substep 5-8, the acetyl group and the benzoyl group protecting the hydroxyl group in the compound of formula X-7 are removed, and the methyl group protecting the carboxylic acid is removed to give the compound of formula XI: [ka] This step can be carried out by utilizing or adapting a known method, but is preferably carried out by, for example, the method shown in Example 63.

[0192] In one embodiment of the present invention, step 5 includes the following substep 5-9 instead of substeps 5-1 to 5-3. [ka]

[0193] <Small process 5-9> Substep 5-9 is a step of preparing a compound of formula X-3 by reacting a compound of formula VI-6 with a compound of formula IX-5. This step can be carried out by utilizing or applying a known method, but is preferably carried out by, for example, the method shown in Example 64.

[0194] This step is preferably carried out in the presence of a Lewis acid. The Lewis acid used in this step is not limited as long as the reaction proceeds, but examples thereof include trimethylsilyl trifluoromethanesulfonate, triisopropylsilyl trifluoromethanesulfonate, and tert-butyldimethylsilyl trifluoromethanesulfonate, and preferred examples include tert-butyldimethylsilyl trifluoromethanesulfonate.

[0195] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include dichloromethane, toluene, trifluoromethylbenzene, and chlorobenzene.

[0196] The reaction temperature in this step is not limited as long as the reaction proceeds, but examples include -40°C to 20°C, preferably -30 to 10°C, more preferably -20 to 5°C, and particularly preferably -10 to 0°C.

[0197] In this step, it is preferable to add 2 to 5 equivalents of the compound represented by formula IX-5 to 1 equivalent of the compound represented by formula VI-6, and it is more preferable to add 3 to 4 equivalents of the compound represented by formula IX-5 to 1 equivalent of the compound represented by formula VI-6.

[0198] It should be noted that steps 1 to 5 and the substeps included therein do not necessarily have to be performed in the order described in this specification. For example, substeps 2-1 and 2-2 in step 2 may be performed prior to step 1. In particular, step 4 may be performed prior to any of steps 1 to 3.

[0199] <2. New method for producing monosaccharides or oligosaccharides> In one aspect of the present invention, a novel method for producing monosaccharides or oligosaccharides is provided, which is useful in a novel purely chemical method for producing biantennary N-glycans having an α2,6-sialic acid structure at the non-reducing end. In the present invention, "oligosaccharide" refers to a saccharide oligomer in which two or more monosaccharides are linked by glycosidic bonds. Furthermore, in the present invention, "monosaccharides" and "oligosaccharides" may each have a protecting group, an activating group, or the like introduced at a specific position.

[0200] The manufacturing method described below is <1> However, the novel method for producing a monosaccharide or oligosaccharide of the present invention is useful in the production of an oligosaccharide of formula XI described in detail above, but is not limited to this application and can be applied to any application. For example, the novel method for producing a monosaccharide or oligosaccharide of the present invention can be used to produce an oligosaccharide of formula XI by a method different from the method for producing an oligosaccharide of formula XI described in detail above. Furthermore, the novel method for producing a monosaccharide or oligosaccharide of the present invention can be used to produce the monosaccharide or oligosaccharide itself, or can be used to produce sugars other than the oligosaccharide of formula XI.

[0201] In one embodiment of the present invention, a compound of formula III-2: [ka] A method for preparing a compound represented by the following formula III-1: [ka] with a strong base of the alkoxide type in the presence of methyl trifluoroacetate to produce a compound of formula III-2.

[0202] Although the acetyl group removal reaction has been reported to be carried out using sodium methoxide in methanol (Org. Biomol. Chem., 2018, 16, 4720-4727), this method can also cause an undesired side reaction, i.e., ring-opening of the phthalimide group. On the other hand, by reacting the compound with a strong alkoxide base in the presence of methyl trifluoroacetate, it is possible to remove the acetyl group while suppressing ring-opening of the phthalimide group.

[0203] The alkoxide-based strong base is not limited as long as the reaction proceeds, and examples thereof include sodium salts, lithium salts, and potassium salts of C1 to C5 alkoxides, and combinations thereof; preferred examples include lithium methoxide, sodium methoxide, potassium methoxide, lithium ethoxide, sodium ethoxide, potassium ethoxide, lithium isopropoxide, sodium isopropoxide, potassium isopropoxide, lithium tert-butoxide, sodium tert-butoxide, potassium tert-butoxide, lithium tert-pentoxide, sodium tert-pentoxide, and potassium tert-pentoxide; particularly preferred examples include sodium methoxide, lithium tert-butoxide, and potassium tert-butoxide.

[0204] The solvent used in this step is not limited as long as the reaction proceeds, and examples thereof include alcoholic solvents such as methanol, ethanol, propanol, and butanol, as well as mixed solvents of tetrahydrofuran, acetonitrile, cyclopentyl methyl ether, toluene, dimethylacetamide, and the like with alcoholic solvents, and preferred examples include methanol or mixed solvents of methanol and tetrahydrofuran.

[0205] The reaction temperature in this step is not limited as long as the reaction proceeds, but examples include -20°C to 80°C, preferably 0°C to 70°C, more preferably 20°C to 65°C, and particularly preferably 40°C to 60°C.

[0206] The method can be suitably carried out, for example, by the method shown in Example 6 or 16.

[0207] In one embodiment of the present invention, a compound of formula III-5: [ka] (wherein X2 is an acetyl group), the method for producing a compound represented by the following formula III-4: [ka] wherein X1 represents a substituent selected from the group consisting of trifluoromethanesulfonyl, nonafluorobutanesulfonyl, 2-nitrobenzenesulfonyl, and 4-nitrobenzenesulfonyl groups, with cesium acetate or tetrabutylammonium acetate to produce a compound of formula III-5: [ka] (wherein X2 is a benzoyl group), the method for producing a compound represented by the following formula III-4: [ka] wherein X1 represents a substituent selected from the group consisting of trifluoromethanesulfonyl, nonafluorobutanesulfonyl, 2-nitrobenzenesulfonyl, and 4-nitrobenzenesulfonyl, with tetrabutylammonium benzoate to produce a compound of formula III-5.

[0208] Although the stereoinversion from glucose to mannose is a known conversion reaction, there have been no reports of conversions in which the protecting group for the hydroxyl group attached to the D-glucopyranoside C-3 of a glucose-glucosamine disaccharide linked by a β-glycosidic bond is a 2-naphthylmethyl (Nap) group. By adopting this method, the stereoinversion from glucose to mannose can be achieved, and a mannose-glucosamine disaccharide skeleton linked by a β-glycosidic bond can be constructed with high yield and high selectivity.

[0209] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include dimethyl sulfoxide, dimethylformamide, dimethylacetamide, N-methylpyrrolidone, N,N-dimethylimidazolidinone, sulfolane, tetrahydrofuran, and acetonitrile, and preferred examples include dimethyl sulfoxide.

[0210] The reaction temperature in this step is not limited as long as the reaction proceeds, but examples include 20 to 80°C, preferably 23 to 70°C, more preferably 26 to 60°C, and particularly preferably 30 to 50°C.

[0211] This method can be preferably carried out by the method shown in Example 9, for example.

[0212] In one embodiment of the present invention, a compound of formula III-8: [ka] A method for preparing a compound represented by the following formula III-7: [ka] by reducing the oxo group attached to the 2-carbon atom of the 2-keto-D-glucopyranoside of a compound represented by formula III-8 to produce a compound represented by formula III-9.

[0213] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include diethyl ether, cyclopentyl methyl ether, tert-butyl methyl ether, diisopropyl ether, dipropyl ether, dibutyl ether, and 1,4-dioxane, and preferably tetrahydrofuran.

[0214] The reaction temperature in this step is not limited as long as the reaction proceeds, and can be, for example, −80° C. to 20° C. As described below, the optimal reaction temperature varies depending on the reducing agent used.

[0215] In one embodiment of the present invention, the oxo group bonded to the carbon atom at the 2-position of 2-keto-D-glucopyranoside in the compound represented by formula III-7 can be substituted with L-selectride, LS-selectride, lithium diisobutyl-tert-butoxyaluminum hydride (LDBBA), or a compound represented by the following formula A: [ka] wherein R3 is a group represented by the following formula: [ka] The reduction is carried out in the presence of a reducing agent selected from the group consisting of compounds represented by the formula (I) and combinations thereof, where R is a di-tert-butylmethylphenoxide or hydride represented by the formula (I), where at least two R are di-tert-butylmethylphenoxides. In this reduction step, for example, when NaBH is used, the stereoselectivity is low (approximately 7:3), making it difficult to efficiently achieve the desired stereoinversion from Gln to Man (Org. Biomol. Chem., 2018, 16, 4720-4727). On the other hand, when the reducing agents listed above are used, the selectivity of the stereoinversion from Gln to Man is significantly improved (93.6:6.4 to 98.1:1.9) compared to when NaBH is used.

[0216] Of the compounds represented by formula A, a compound in which three R3 are di-tert-butylmethylphenoxide can be obtained, for example, by adding dibutylhydroxytoluene (885.41 mg, 4.02 mmol) to a tetrahydrofuran suspension (2 mL) of lithium aluminum hydride (50.0 mg, 1.32 mmol) at 0° C., followed by stirring at 25° C. Of the compounds represented by formula A, a compound in which two R3 are di-tert-butylmethylphenoxide can be obtained in a similar manner by using 2 molar equivalents of dibutylhydroxytoluene for 1 molar equivalent of lithium aluminum hydride.

[0217] As described above, the reaction temperature in this step is not limited as long as the reaction proceeds, but when L-selectride, LS-selectride, or LDBBA is used as the reducing agent, the reaction temperature is preferably −80° C. to −20° C., more preferably −80° C. to −30° C., even more preferably −80° C. to −40° C., and particularly preferably −80° C. to −50° C. When a compound represented by formula A is used as the reducing agent, the reaction temperature is preferably −20° C. to 20° C., more preferably −15° C. to 15° C., and particularly preferably −10° C. to 10° C. Therefore, the compound represented by formula A is particularly preferred as the reducing agent used in this step, in that the reaction proceeds at a temperature that is easier to handle.

[0218] This method can be preferably carried out, for example, by the method shown in Example 19.

[0219] In one embodiment of the present invention, a compound of formula III-9: [ka] The method for preparing a compound represented by the following formula III-8: [ka] In a compound represented by formula III-10, the hydroxyl group attached to the 2-carbon atom of the D-mannopyranoside is protected with a benzyl group to produce a compound represented by formula III-9.

[0220] The benzyl group is protected in the presence of lithium tert-butoxide or lithium tert-amoxide, which prevents ring-opening of the phthalimide. Furthermore, this method is safer and easier to scale up than the typical method using sodium hydride.

[0221] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include dimethylacetamide, dimethylformamide, N-methylpyrrolidone, and N,N-dimethylimidazolidinone, and preferably dimethylacetamide.

[0222] The reaction temperature in this step is not limited as long as the reaction proceeds, but may be, for example, -20 to 100°C, preferably -15 to 70°C, and particularly preferably -10 to 50°C.

[0223] The method can be suitably carried out, for example, by the method shown in Example 11 or 21.

[0224] In one embodiment of the present invention, a compound of formula V-5: [ka] A method for producing a compound represented by the following formula V-4: [ka] with a strong base of the alkoxide type in the presence of methyl trifluoroacetate to produce a compound of formula V-5.

[0225] Although it is known that deprotection of the acetyl group is generally carried out using sodium methoxide in methanol, this can also result in an undesired side reaction, i.e., ring-opening of the phthalimide group. On the other hand, by using a method in which the acetyl group is reacted with a strong alkoxide base in the presence of methyl trifluoroacetate, it is possible to remove the acetyl group while suppressing ring-opening of the phthalimide group.

[0226] The alkoxide-based strong base is not limited as long as the reaction proceeds, and examples thereof include sodium salts, lithium salts, and potassium salts of C1 to C5 alkoxides, and combinations thereof; preferred examples include lithium methoxide, sodium methoxide, potassium methoxide, lithium ethoxide, sodium ethoxide, potassium ethoxide, lithium isopropoxide, sodium isopropoxide, potassium isopropoxide, lithium tert-butoxide, sodium tert-butoxide, potassium tert-butoxide, lithium tert-pentoxide, sodium tert-pentoxide, and potassium tert-pentoxide; particularly preferred examples include sodium methoxide, lithium tert-butoxide, and potassium tert-butoxide.

[0227] The solvent used in this step is not limited as long as the reaction proceeds, and examples thereof include alcoholic solvents such as methanol, ethanol, propanol, and butanol, as well as mixed solvents of tetrahydrofuran, acetonitrile, cyclopentyl methyl ether, toluene, dimethylacetamide, and the like with alcoholic solvents, and preferred examples include methanol or mixed solvents of methanol and tetrahydrofuran.

[0228] The reaction temperature in this step is not limited as long as the reaction proceeds, but examples include -20°C to 80°C, preferably 0°C to 70°C, more preferably 15°C to 65°C, and particularly preferably 30°C to 60°C.

[0229] This method can be suitably carried out, for example, by the method shown in Example 29.

[0230] In one embodiment of the present invention, a compound of formula II-5: [ka] or a compound of formula II-8: [ka] A method for producing a compound represented by the formula: In fluorous alcohol and water, the following formula II-4: [ka] or a compound of formula II-7: [ka] with λ3-iodane to produce a compound represented by formula II-5 or a compound represented by formula II-8.

[0231] In the present invention, "λ3-iodane" refers to a trivalent hypervalent iodine compound. Compared with conventional deprotection methods, the use of λ3-iodane not only increases the yield of the compound represented by Formula II-5 or Formula II-8, but also allows the reaction to be carried out under milder reaction conditions. Furthermore, because the reaction proceeds with a slight excess of λ3-iodane, purification of the deprotected product is easier and operational safety is higher than with conventional deprotection methods that use, for example, an excess of ammonium cerium (IV) nitrate.

[0232] In one embodiment, the λ-iodane has the formula R 4 -I(OR 5 ) 2 (wherein R 4 is an unsubstituted or substituted phenyl group, and R 5 is selected from the group consisting of H, acetyl, trifluoroacetyl, tosyl, methanesulfonyl, and combinations thereof. As defined in the formula above, R 4may be a "substituted phenyl group," and examples of the substituent include a linear or branched saturated or unsaturated hydrocarbon group, an oxygen-containing group (alkoxy, ester, etc.), a nitrogen-containing group (cyano, azide, etc.), and a halogen (e.g., fluorine, chlorine, bromine, iodine), with a hydrocarbon group, an oxygen-containing substituent, or a halogen being more preferred. When these substituents contain carbon, for example, those having 1 to 5 carbon atoms or those having 1 to 3 carbon atoms can be suitably used. Specific examples of λ3-iodane include, but are not limited to, [bis(trifluoroacetoxy)iodo]benzene (PIFA), [hydroxy(tosyloxy)iodo]benzene (HTIB), (diacetoxyiodo)benzene (PIDA), [bis(trifluoroacetoxy)iodo]pentafluorobenzene, and [hydroxy(methanesulfonyloxy)iodo]benzene.

[0233] The amount of λ3-iodane can be appropriately set, but may be, for example, about 0.1 to 10 equivalents, about 0.5 to 7 equivalents, or about 1 to 5 equivalents relative to the compound represented by formula II-4 or the compound represented by formula II-7, and preferably about 1 to 3 equivalents.

[0234] In the present invention, "fluorous alcohol" refers to a fluorine-containing alcohol compound in which all carbon atoms except the carbon bonded to the alcohol have fluorine. As long as fluorine substitution is permitted, it is preferable for the fluorous alcohol to have more fluorine atoms. Fluorous alcohols include, but are not limited to, fluorous aliphatic alcohols. The hydrocarbon moiety in the fluorous aliphatic alcohol may be saturated or unsaturated, linear or branched, or cyclic. Examples of fluorous aliphatic alcohols include fluorous C2-C8 aliphatic alcohols, preferably fluorous C2-C5 aliphatic alcohols, and more preferably fluorous C2-C3 aliphatic alcohols. Specific examples of fluorous alcohols include, but are not limited to, hexafluoro-2-propanol (HFIP), 2,2,2-trifluoroethanol (TFE), 2,2,3,3,4,4,5,5-octafluoro-1-pentanol, nonafluoro-tert-butyl alcohol, and combinations thereof.

[0235] Furthermore, preferred combinations of fucolic alcohol and λ3-iodane can provide the deprotected product in higher yields. Those skilled in the art can select appropriate combinations, but examples include, but are not limited to, PIFA preferably used in combination with hexafluoro-2-propanol (HFIP), 2,2,2-trifluoroethanol (TFE), nonafluoro-tert-butyl alcohol, HTIB preferably used in combination with HFIP, TFE, 2,2,3,3,4,4,5,5-octafluoro-1-pentanol, [bis(trifluoroacetoxy)iodo]pentafluorobenzene preferably used in combination with hexafluoro-2-propanol (HFIP), and [hydroxy(methanesulfonyloxy)iodo]benzene preferably used in combination with hexafluoro-2-propanol (HFIP).

[0236] The amount of fluorous alcohol can be appropriately set from the viewpoint of achieving a high yield of the product, and may be, for example, about 1.0 equivalent or more, about 1.5 equivalents or more, about 2.0 equivalents or more, or 2.5 equivalents or more in molar ratio relative to the compound represented by formula II-4 or the compound represented by formula II-7, and may be about 15 or less, about 10 or less, about 8 or less, or about 5 or less in volume ratio relative to the compound represented by formula II-4 or the compound represented by formula II-7.

[0237] This step is carried out in the presence of the fluorous alcohol and water. The amount of water can be appropriately determined from the viewpoint of achieving a high yield of the product, and may be, for example, about 1.0 equivalent or more, about 1.5 equivalents or more, about 2.0 equivalents or more, or about 2.5 equivalents or more in molar ratio relative to the compound represented by Formula II-4 or II-7, and about 10 or less, about 8 or less, about 5 or less, or about 3 or less in volume ratio relative to the compound represented by Formula II-4 or II-7.

[0238] In this step, a "solvent" may be further added to the fluorous alcohol and water. The solvent may be selected from the group consisting of, but not limited to, dichloromethane, toluene, (trifluoromethyl)benzene, and combinations thereof. The type of solvent used may be appropriately selected depending on the λ3-iodane used, etc., to achieve a high yield of the product. The amount of solvent may also be appropriately set to achieve a high yield of the product, and may be, for example, about 0.5 to 50, about 1 to 20, or about 2 to 10 by volume relative to the compound represented by Formula II-4 or the compound represented by Formula II-7.

[0239] In this step, an "additive" may be further added to the fluorous alcohol and water. The additive is preferably selected from the group consisting of sodium dihydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, and combinations thereof. Since the acidity may increase as the 4-methoxyphenylation reaction proceeds, adding an additive such as sodium dihydrogen phosphate can achieve a higher product yield, particularly when using λ3-iodane (e.g., HTIB), which by-produces a strongly acidic acid. The amount of the additive can also be appropriately selected to achieve a high product yield; for example, it can be about 0.5 to 8 equivalents, about 1 to 6 equivalents, or about 1.5 to 5 equivalents relative to the compound represented by Formula II-4 or the compound represented by Formula II-7.

[0240] When (diacetoxyiodo)benzene (PIDA) is used as the λ3-iodane, it is preferable to add trifluoroacetic acid (TFA) to obtain the deprotected product in higher yield.

[0241] In this process, λ3-iodane acts as an oxidizing agent on the 4-methoxyphenyl (X group) in the compound of Formula II-4 or II-7 in the presence of a fluorous alcohol and water, resulting in the elimination of the OX group from the compound of Formula II-4 or II-7. Water (HO) then adds to the oxidized 4-methoxyphenoxy (OX group) from the compound of Formula II-4 or II-7. This oxidizing action of λ3-iodane is generally achieved by stirring, refluxing, or otherwise treating a solution containing the compound of Formula II-4 or II-7 and λ3-iodane in a fluorous alcohol and water. Therefore, this process is simple and easily scaled up. The product, the compound of Formula II-5 or II-8, can be purified or isolated by any purification method known to those skilled in the art, such as crystallization or chromatography.

[0242] The temperature when reacting the compound represented by Formula II-4 or the compound represented by Formula II-7 with λ3-iodane is preferably from about −20° C. to the boiling point of the fluorous alcohol (e.g., about 58° C. for hexafluoro-2-propanol (HFIP) and about 78° C. for 2,2,2-trifluoroethanol (TFE)) or lower, and can be, for example, about −20 to 60° C., about 0 to 60° C., or about 10 to 30° C. Another advantage is that the demethoxyphenylation reaction proceeds even at room temperature (15 to 30° C.), eliminating the need for cooling or heating.

[0243] The reaction time can be appropriately set to obtain the product in high yield.

[0244] The method can be suitably carried out, for example, by the method shown in Example 33 or 38.

[0245] In one embodiment of the present invention, a compound of formula II-6: [ka] or a compound of formula II-9: [ka] A method for preparing a compound represented by the following formula II-5: [ka] or a compound of formula II-8: [ka] in the presence of N-methylimidazole, with a compound of the formula: [ka] with 2,2,2-trifluoro-N-phenylacetimidoyl chloride (TFPC) of formula II-6 to produce a compound of formula II-9.

[0246] The use of N-methylimidazole as a base in this step significantly improves the yield. For example, when producing the compound of formula II-6 from the compound of formula II-5, the yield is about 36% when potassium carbonate is used under certain conditions, but the yield is significantly improved to about 76% when N-methylimidazole is used. Since TFPC is an expensive reagent, improving the yield of this step is extremely beneficial for commercial production.

[0247] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include dichloromethane, toluene, ethyl acetate, acetonitrile, and tetrahydrofuran, and preferably dichloromethane.

[0248] The reaction temperature in this step is not limited as long as the reaction proceeds, but is preferably -20 to 40°C, more preferably -10 to 35°C, and particularly preferably 0 to 30°C.

[0249] This step is preferably carried out in the presence of a dehydrating agent. The dehydrating agent used in this step is not limited as long as the reaction proceeds, but examples thereof include molecular sieves, and preferably molecular sieves 4A powder with a powder particle size of 10 μm or less.

[0250] In this method, the compound represented by Formula II-6 produced can be isolated and purified by column or recrystallization. Examples of column-based isolation and purification include isolation and purification using silica gel as the stationary phase and dichloromethane or a toluene-ethyl acetate mixed solvent system as the mobile phase. On the other hand, examples of isolation and purification by recrystallization include crystallization from a mixed solvent system of diisopropyl ether and heptane. Recrystallization can be preferably carried out using seed crystals of the compound represented by Formula II-6. When using seed crystals, the compound can be obtained, for example, by inoculating the seed crystals into a diisopropyl ether solution, confirming the precipitation of crystals, and then adding heptane dropwise.

[0251] The method can be suitably carried out, for example, by the method shown in Example 34 or 39.

[0252] In one embodiment of the present invention, a compound of formula VI-5: [ka] A method for preparing a compound represented by the following formula VI-4: [ka] with a strong base of the alkoxide type in the presence of methyl trifluoroacetate to produce a compound of formula VI-5.

[0253] Although it is known that deprotection of the acetyl group is generally carried out using sodium methoxide in methanol, this can also result in an undesired side reaction, i.e., ring-opening of the phthalimide group. On the other hand, by using a method in which the acetyl group is reacted with a strong alkoxide base in the presence of methyl trifluoroacetate, it is possible to remove the acetyl group while suppressing ring-opening of the phthalimide group.

[0254] The alkoxide-based strong base is not limited as long as the reaction proceeds, and examples thereof include sodium salts, lithium salts, and potassium salts of C1 to C5 alkoxides, and combinations thereof; preferred examples include lithium methoxide, sodium methoxide, potassium methoxide, lithium ethoxide, sodium ethoxide, potassium ethoxide, lithium isopropoxide, sodium isopropoxide, potassium isopropoxide, lithium tert-butoxide, sodium tert-butoxide, potassium tert-butoxide, lithium tert-pentoxide, sodium tert-pentoxide, and potassium tert-pentoxide; particularly preferred examples include sodium methoxide, lithium tert-butoxide, and potassium tert-butoxide.

[0255] The solvent used in this step is not limited as long as the reaction proceeds, and examples thereof include alcoholic solvents such as methanol, ethanol, propanol, and butanol, as well as mixed solvents of tetrahydrofuran, acetonitrile, cyclopentyl methyl ether, toluene, dimethylacetamide, and the like with alcoholic solvents, and preferred examples include methanol or mixed solvents of methanol and tetrahydrofuran.

[0256] The reaction temperature in this step is not limited as long as the reaction proceeds, but examples include -20°C to 80°C, preferably 0°C to 70°C, more preferably 15°C to 65°C, and particularly preferably 30°C to 60°C.

[0257] This method can be suitably carried out, for example, by the method shown in Example 41.

[0258] In one embodiment of the present invention, a compound of formula VII-3: [ka] The present invention provides a method for preparing a compound of formula VII-3, which comprises contacting a solvent in which the compound of formula VII-3 is dissolved with silica gel to perform solid-phase extraction of the compound of formula VII-3.

[0259] The unreacted compound of formula VII-2, which can be used as a raw material for producing the compound of formula VII-3, and the eliminated benzaldehyde are not adsorbed onto silica gel, so the compound of formula VII-3 can be efficiently purified by this process.

[0260] Examples of solvents for dissolving the compound represented by formula VII-3 include toluene, heptane, dichloromethane, chloroform, and combinations thereof. Preferred examples include toluene, dichloromethane, chloroform, and combinations thereof. Particularly preferred examples include, but are not limited to, toluene.

[0261] The amount of silica gel used in this step can be, for example, 2 to 5 times the amount of the raw materials, preferably 2 to 4 times the amount of the raw materials, and more preferably about 3 times the amount of the raw materials.

[0262] In this step, the solvent for eluting the compound of formula VII-3 adsorbed on the silica gel is not particularly limited as long as it does not dissolve the silica gel and can elute the target compound. Examples of the solvent include cyclopentyl methyl ether, ethyl acetate, and tert-butyl methyl ether.

[0263] This method can be suitably carried out, for example, by the method shown in Example 46.

[0264] In one embodiment of the present invention, a compound of formula VIII-4: [ka] A method for preparing a compound represented by the following formula VIII-3: [ka] in the presence of N-methylimidazole, with a compound of the formula: [ka] with 2,2,2-trifluoro-N-phenylacetimidoyl chloride (TFPC) of formula VIII-4 to produce a compound of formula VIII-4.

[0265] Compared with the case of using potassium carbonate as the base in this step, the use of N-methylimidazole allows for a reduction in the equivalent amount of TFPC, and still allows for the production of the target compound in high yield. Since TFPC is an expensive reagent, improving the yield of this step is extremely beneficial for commercial production.

[0266] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include dichloromethane, toluene, ethyl acetate, acetonitrile, and tetrahydrofuran, and preferably dichloromethane.

[0267] The reaction temperature in this step is not limited as long as the reaction proceeds, but is preferably -20 to 40°C, more preferably -10 to 35°C, and particularly preferably 0 to 30°C.

[0268] This step is preferably carried out in the presence of a dehydrating agent. The dehydrating agent used in this step is not limited as long as the reaction proceeds, but examples thereof include molecular sieves, and preferably molecular sieves 4A powder with a powder particle size of 10 μm or less.

[0269] This step can be preferably carried out by the method shown in Example 49, for example.

[0270] In one embodiment of the present invention, a compound of formula IX-1: [ka] A method for producing a compound represented by the following formula VIII-5: [ka] and a compound represented by the following formula VII-3: [ka] by α-2,6-glycosidic bonding to form a compound represented by formula IX-1.

[0271] Selective α-2,6-glycosidic coupling of N-acetylneuraminic acid derivatives and galactose derivatives has been difficult. For example, a method for synthesizing disaccharides by reacting a compound represented by formula VIII-4 with a compound represented by formula VII-3 has been reported (J. Org. Chem., 2016, 81, 10600-10616). However, the reaction was difficult to reproduce, and the desired yield and selectivity could not be achieved. Furthermore, this reaction had issues such as a decrease in selectivity with increasing scale, a narrow reaction temperature tolerance, and a significant effect of reaction heat. Because the compound represented by formula VIII-4, one of the raw materials for this reaction, is very expensive, the low reproducibility, yield, and selectivity of this reaction pose a major problem, especially for commercial production, where scale-up is required. On the other hand, when the compound represented by formula VIII-5, which has a tert-butoxycarbonyl group added thereto, is used as the starting compound instead of the compound represented by formula VIII-4, high selectivity for α-2,6-glycosidic bonds (α:β=93:7) can be achieved with good reproducibility and improved yield. Furthermore, the temperature tolerance is broadened, and high reproducibility, yield, and selectivity can be achieved even when scaled up. This is extremely beneficial for commercial production.

[0272] This step is preferably carried out in the presence of a Lewis acid. The Lewis acid used in this step is not limited as long as the reaction proceeds, but examples thereof include trimethylsilyl trifluoromethanesulfonate, triisopropylsilyl trifluoromethanesulfonate, and tert-butyldimethylsilyl trifluoromethanesulfonate, and preferably trimethylsilyl trifluoromethanesulfonate.

[0273] The solvent used in this step is not limited as long as the reaction proceeds, but examples thereof include diisopropyl ether, tert-butyl methyl ether, diethyl ether, dibutyl ether, dipropyl ether, 1,4-dioxane, dichloromethane, 1,2-dichloroethane, toluene, chlorobenzene, trifluoromethylbenzene, propionitrile, and acetonitrile, and preferred examples include cyclopentyl methyl ether.

[0274] The reaction temperature in this step is not limited as long as the reaction proceeds, but examples thereof include -78°C to 0°C, preferably -78 to -20°C, more preferably -78 to -30°C, and particularly preferably -78 to -40°C.

[0275] In this step, it is preferable to add 1 to 3 equivalents of the compound represented by formula VII-3 to 1 equivalent of the compound represented by formula VIII-5, and it is more preferable to add 1.4 to 2 equivalents of the compound represented by formula VII-3 to 1 equivalent of the compound represented by formula VIII-5.

[0276] This step can be carried out, for example, by adding a mixed solution of the compound represented by Formula VIII-5 and the compound represented by Formula VII-3 (preferably a cyclopentyl methyl ether mixed solution) dropwise to a solution containing a Lewis acid (preferably a cyclopentyl methyl ether solution) for a long period of time, or by adding a solution of the compound represented by Formula VIII-5 (preferably a cyclopentyl methyl ether solution) dropwise to a solution containing a Lewis acid and the compound represented by Formula VII-3 (preferably a cyclopentyl methyl ether solution) for a long period of time. The addition time is not limited as long as the reaction proceeds, but is, for example, 30 minutes to 5 hours, preferably 1 to 4 hours, more preferably 2 to 3.5 hours, and particularly preferably about 3 hours.

[0277] This method can be suitably carried out, for example, by the method shown in Example 51.

[0278] In one embodiment of the present invention, a compound of formula IX-1: [ka] The present invention provides a method for preparing a compound of formula IX-1, which comprises contacting a solvent in which the compound of formula IX-1 is dissolved with silica gel to perform solid phase extraction of the compound of formula IX-1.

[0279] In the production process of the compound of formula IX-1, N-phenyltrifluoroacetamide, a by-product of the glycosylation reaction, and other trace impurities in the toluene solvent that are not adsorbed by silica gel are not adsorbed by silica gel, so the compound of formula IX-1 can be efficiently purified by this process.

[0280] Examples of solvents for dissolving the compound represented by formula IX-1 include toluene, heptane, dichloromethane, chloroform, and combinations thereof. Preferred examples include toluene, dichloromethane, chloroform, and combinations thereof. Particularly preferred examples include, but are not limited to, toluene.

[0281] The amount of silica gel used in this step can be, for example, 2 to 5 times the amount of the raw materials, preferably 2 to 4 times the amount of the raw materials, and more preferably about 3.5 times the amount of the raw materials.

[0282] In this step, the solvent for eluting the compound represented by formula IX-1 adsorbed on the silica gel is not particularly limited as long as it does not dissolve the silica gel and can elute the target compound. Examples of the solvent include ethyl acetate, cyclopentyl methyl ether, and tert-butyl methyl ether, and preferably ethyl acetate.

[0283] This method can be suitably carried out, for example, by the method shown in Example 51.

[0284] In one embodiment of the present invention, a compound of formula IX-3: [ka] The present invention provides a method for preparing a compound of formula IX-3, which comprises contacting a solvent in which the compound of formula IX-3 is dissolved with silica gel to perform solid phase extraction of the compound of formula IX-3.

[0285] In one embodiment of the process for producing the compound of formula IX-3, by-products such as the diacetyl form of the compound of formula VII-3, which is produced by acetylating the compound of formula VII-3 used in excess in the upstream glycosylation reaction, are not adsorbed onto silica gel, and therefore the compound of formula IX-3 can be efficiently purified by this process.

[0286] Examples of solvents for dissolving the compound represented by formula IX-3 include toluene, heptane, dichloromethane, chloroform, and combinations thereof. Preferred examples include toluene, dichloromethane, chloroform, and combinations thereof. Particularly preferred examples include, but are not limited to, toluene.

[0287] The amount of silica gel used in this step can be, for example, 2 to 5 times the amount of the raw materials, preferably 2 to 4 times the amount of the raw materials, and more preferably about 3.5 times the amount of the raw materials.

[0288] In this step, the solvent for eluting the compound represented by formula IX-3 adsorbed on the silica gel is not particularly limited as long as it does not dissolve the silica gel and can elute the target compound. Examples of the solvent include ethyl acetate, cyclopentyl methyl ether, and tert-butyl methyl ether, and preferably ethyl acetate.

[0289] This method can be suitably carried out, for example, by the method shown in Example 53.

[0290] Each of the novel methods for producing monosaccharides or oligosaccharides described above can be used in the method for producing the oligosaccharide represented by formula XI. Accordingly, in one aspect of the present invention, there is provided a method for producing the oligosaccharide represented by formula XI, including the novel method for producing the monosaccharide or oligosaccharide. In this context, the method for producing the oligosaccharide represented by formula XI can be described in the present specification. <1> The method for producing the oligosaccharide of formula XI may be the same as or different from the method detailed in the preceding paragraph. <1> , the method may be different from the method detailed in the preceding paragraphs by utilizing or adapting known methods for producing oligosaccharides of formula XI in addition to the novel methods for producing monosaccharides or oligosaccharides described above, by using or adapting conventional methods for monosaccharide or oligosaccharide production, and / or by adapting conventional methods for monosaccharide or oligosaccharide production described herein. <1> This can be achieved by partially utilizing or applying the method detailed in .

[0291] <3. New intermediates> In one aspect of the present invention, novel intermediates useful in a novel purely chemical method for producing biantennary N-glycans having an α2,6-sialic acid structure at the non-reducing end are provided. The intermediates described below are useful for producing the oligosaccharide represented by formula XI described in detail above, but are not limited to this use and can be applied to any other uses. For example, the novel intermediates of the present invention can also be used for producing saccharides other than the oligosaccharide represented by formula XI.

[0292] In one embodiment of the present invention, a compound of formula III-6: [ka] The compound is provided as follows: <1> It can be produced by the method described in .

[0293] In one embodiment of the present invention, a compound of formula III-10: [ka] The compound is provided as follows: <1> It can be produced by the method described in .

[0294] In one embodiment of the present invention, a compound of formula V-1: [ka] The compound is provided as follows: <1> It can be produced by the method described in .

[0295] In one embodiment of the present invention, a compound of formula V-2: [ka] The compound is provided as follows: <1> It can be produced by the method described in .

[0296] In one embodiment of the present invention, the compound of formula V-3: [ka] The compound is provided as follows: <1> It can be produced by the method described in .

[0297] In one embodiment of the present invention, a compound of formula II-6: [ka] The compound is provided as follows: <1> It can be produced by the method described in .

[0298] In one embodiment of the present invention, a compound of formula VI-A: [ka] (Wherein R1 is [ka] The compound in which R1 is a 2,2,2-trichloroethoxycarbonylamino group is <1> The compound in which R1 is a phthalimido group corresponds to the compound represented by formula VI-1 mentioned above. <1> These compounds correspond to the compounds of formula VI-4 mentioned above. <1> On the other hand, the compound in which R1 is NHAc can be obtained, for example, in Example 44, as an intermediate (compound represented by formula VI-7) in the preparation of the compound represented by formula VI-6.

[0299] In one embodiment of the present invention, the compound of formula VI-2: [ka] The compound is provided as follows: <1> It can be produced by the method described in .

[0300] In one embodiment of the present invention, a compound of formula VI-B: [ka] (Wherein R2 is [ka] The compound in which R2 is a 2,2,2-trichloroethoxycarbonylamino group is <1> The compound in which R1 is a phthalimido group corresponds to the compound represented by formula VI-3 mentioned above. <1> The compound corresponds to the compound represented by formula VI-5 mentioned above, and the compound in which R1 is an acetylamino group is <1> These compounds correspond to the compounds of formula VI-6 mentioned above. <1> It can be produced by the method described in .

[0301] In one embodiment of the present invention, a compound of formula VIII-5: [ka] The compound is provided as follows: <1> It can be produced by the method described in .

[0302] In one embodiment of the present invention, a compound of formula X-1: [ka] The compound is provided as follows: <1> It can be produced by the method described in .

[0303] In one embodiment of the present invention, a compound of formula X-2: [ka] The compound is provided as follows: <1> It can be produced by the method described in .

[0304] In one embodiment of the present invention, a compound of formula X-3: [ka] The compound is provided as follows: <1> It can be produced by the method described in .

[0305] In one embodiment of the present invention, a compound of formula X-4: [ka] The compound is provided as follows: <1> It can be produced by the method described in .

[0306] In one embodiment of the present invention, a compound of formula X-5: [ka] The compound is provided as follows: <1> It can be produced by the method described in .

[0307] In one embodiment of the present invention, a compound of formula X-6: [ka] The compound is provided as follows: <1> It can be produced by the method described in .

[0308] In one embodiment of the present invention, a compound of formula X-7: [ka] The compound is provided as follows: <1> It can be produced by the method described in .

[0309] <4. New method for producing glycoproteins, etc.> In one aspect of the present invention, there is provided a novel purely chemical method for producing a biantennary N-glycan having an α2,6-sialic acid structure at the non-reducing end (i.e., an oligosaccharide represented by formula XI), or a novel method for producing a glycoprotein (particularly, a glycosylated antibody or an Fc region-containing molecule thereof, or an antibody-drug conjugate) using the novel method for producing the N-glycan, including a novel method for producing a monosaccharide or oligosaccharide. As described in detail below, the oligosaccharide represented by formula XI obtained by the production method of the present invention can be used for the production of glycoproteins (particularly, a glycosylated antibody or an Fc region-containing molecule thereof, or an antibody-drug conjugate) (WO2019 / 065964, WO2020 / 050406, etc.), but it can also be used for other purposes, including but not limited to these.

[0310] Recently, a method for remodeling heterogeneous antibody glycans by enzymatic reactions to uniformly introduce functionalized glycans has been reported (ACS Chem. Biol. 2012, 7, 110-122, ACS Med. Chem. Lett. 2016, 7, 1005-1008). Using this glycan remodeling technique, attempts have also been made to site-specifically introduce drugs and synthesize homogeneous antibody-drug conjugates (ADCs) (Bioconjugate Chem. 2015, 26, 2233-2242, Angew. Chem. Int. Ed. 2016, 55, 2361-2367, US2016361436).

[0311] In glycan remodeling, first, a hydrolase is used to remove heterogeneous glycans attached to a protein (e.g., an antibody) leaving only the terminal N-acetylglucosamine (GlcNAc) to prepare a homogeneous protein portion with GlcNAc attached (hereafter referred to as the "acceptor molecule"). Next, a separately prepared glycan of your choice (hereafter referred to as the "donor molecule") is prepared, and this acceptor molecule and donor molecule are linked using a glycosyltransferase. This allows the synthesis of a homogeneous glycoprotein with a desired glycan structure.

[0312] In one embodiment of the present invention, the oligosaccharide represented by formula XI produced by the novel production method of the present invention can be used as a donor molecule for synthesizing the above-mentioned homogeneous glycoprotein (particularly, a glycosylated antibody or its Fc region-containing molecule) by activating its terminal structure.

[0313] In the present invention, the term "glycan" refers to a structural unit in which two or more monosaccharides are linked by glycosidic bonds. Specific monosaccharides and glycans may be represented by abbreviations such as "GlcNAc-" or "SG-." When these abbreviations are used in structural formulae, the oxygen or nitrogen atom at the reducing end that forms a glycosidic bond with another structural unit is not included in the abbreviation representing the glycan, unless otherwise defined.

[0314] In the present invention, unless otherwise specified, monosaccharides, which are the basic units of sugar chains, are described by assuming that the carbon atom bonded to the oxygen atom constituting the ring and directly bonded to a hydroxy group (or an oxygen atom belonging to a glycosidic bond) in the ring structure is at position 1 (position 2 only in sialic acid). The names of the example compounds are given based on the entire chemical structure, and this rule does not necessarily apply.

[0315] In the present invention, when a sugar chain is represented by a symbol (e.g., SG, GlcNAc, etc.), unless otherwise defined, the symbol includes the carbon atom at the reducing end, and does not include the N or O attributable to the N- or O-glycosidic bond.

[0316] SGP (α2,6-SGP) is an abbreviation for sialyglycopeptide and is a representative N-linked glycopeptide. In this specification, the sugar chain portion of SGP is abbreviated as SG, and the sugar chain lacking one GlcNAc at the reducing end of SG is abbreviated as SG(10). SG(10) is synonymous with the oligosaccharide represented by formula XI.

[0317] In the present invention, the term "glycoprotein" refers to a protein having a sugar chain bound to a portion of the amino acids constituting the protein. In the present invention, the term "glycoprotein" includes, but is not limited to, a sugar chain-remodeling antibody or a molecule containing the Fc region thereof.

[0318] In one embodiment of the present invention, the "glycoprotein" is a glycan-remodeling antibody or an Fc region-containing molecule thereof. Accordingly, in one embodiment of the present invention, there is provided a method for producing a glycan-remodeling antibody or an Fc region-containing molecule thereof, which comprises the novel production method of the present invention and further comprises the steps of obtaining a glycan donor molecule containing N-acetylglucosamine (GlcNAc) with an activated reducing end from the obtained oligosaccharide represented by formula XI, and reacting the glycan donor molecule with an acceptor molecule that is an antibody or an Fc region-containing molecule thereof having a core GlcNAc optionally containing fucose as the N297-linked glycan. In the present invention, "core GlcNAc" refers to the GlcNAc at the reducing end of the N297-linked glycan.

[0319] In the present invention, the term "sugar chain donor molecule" refers to a molecule that plays a role in adding a sugar chain to an acceptor molecule.

[0320] In the present invention, "GlcNAc with an activated reducing end" refers to GlcNAc with an excellent leaving group at the reducing end. Examples of "GlcNAc with an activated reducing end" include oxazolinated GlcNAc, halogenated GlcNAc, phenylated GlcNAc, para-nitrophenylated GlcNAc, and GlcNAc with dimethoxytriazine at the anomeric position (4,6-dimethoxy-1,3,5-triazin-2-yl glycoside (DMT sugar)), and preferred examples include oxazolinated GlcNAc and fluorinated GlcNAc. For example, a sugar chain donor molecule containing oxazolinated GlcNAc can be represented by the following formula XII: [ka] The compound is represented by the formula (also called "SG(10)-Ox").

[0321] In the present invention, the non-reducing end of a "sugar chain donor molecule" may be chemically modified. The group added by chemical modification, the chemical reaction involved in the chemical modification, the functional group on the sugar chain donor to be chemically modified, etc. can be appropriately selected depending on the purpose. In one embodiment of the present invention, the sugar chain donor molecule may be chemically modified by reacting the carboxyl group at the 2-position of the sialic acid located at its non-reducing end with a compound having an amino group at its terminal to form an amide bond. The compound having an amino group at its terminal is not particularly limited, but examples thereof include compounds having an azide group at the other terminal, and preferably compounds represented by the following formula: [ka] Examples of suitable amines include 11-azido-3,6,9-trioxaundecan-1-amines represented by the following formula:

[0322] In one embodiment of the present invention, the "sugar chain donor molecule" is an azido-sugar chain oxazoline compound in which the reducing end is oxazolinated and an azide group-containing group is added to the non-reducing end. The azido-sugar chain oxazoline compound can be synthesized according to the method described in WO2018 / 003983. As an example, the synthesis method of [N3-PEG(3)]2-SG(10)-Ox (compound 1-10 described in WO2018 / 003983) is shown in the following scheme.

[0323] [ka]

[0324] In the present invention, "acceptor molecule" refers to a molecule that receives a sugar chain from a sugar chain donor molecule. The "acceptor molecule" is not particularly limited as long as it is a molecule that can receive a sugar chain from a sugar chain donor molecule, but preferred examples include glycosylated antibodies in which most of the sugar chain has been cleaved from an antibody, or molecules containing the Fc region thereof, and more preferred examples include antibodies having a core GlcNAc or molecules containing the Fc region thereof. The sugar chain that is partially cleaved in a glycosylated antibody is an N-linked sugar chain or an O-linked sugar chain, and preferably an N-linked sugar chain.

[0325] N-linked glycans are bound to amino acid side chains of antibodies via N-glycosidic bonds, and O-linked glycans are bound to amino acid side chains of antibodies via O-glycosidic bonds.

[0326] The antibody serving as the "acceptor molecule" is preferably an IgG, and more preferably an IgG1, IgG2, or IgG4.

[0327] IgG has a well-conserved N-linked glycan (hereinafter referred to as "Asn297-linked glycan" or "N297-linked glycan") at the 297th asparagine residue (hereinafter referred to as "Asn297 or N297") in the Fc region of its heavy chain, and it is known to contribute to the activity and kinetics of antibody molecules (Eon-Duval, A. et al., Biotechnol. Prog. 2012, 28, 608-622; Sanglier-Cianferani, S., Anal. Chem. 2013, 85, 715-736).

[0328] The amino acid sequence in the constant region of IgG is well conserved, and in a report by Edelman et al. (Proc. Natl. Acad. Sci. USA, 63, 78-85, (1969)), each amino acid is identified by an EU number (EU INDEX). For example, Asn297, to which N-linked glycosylation is added in the Fc region, corresponds to position 297 in the EU numbering. Even if the actual amino acid position changes due to molecular fragmentation or region deletion, the amino acid can be uniquely identified by indicating it by EU numbering.

[0329] In one embodiment of the present invention, the "acceptor molecule" is an antibody having a core GlcNAc as the N297-linked glycan or a molecule containing an Fc region thereof. The core GlcNAc as the N297-linked glycan may have another monosaccharide or glycan attached thereto, for example, fucose attached thereto. Thus, in one embodiment of the present invention, the "acceptor molecule" is an antibody having a core GlcNAc as the N297-linked glycan, optionally with fucose attached thereto, or a molecule containing an Fc region thereof.

[0330] In the step of reacting a glycan donor molecule with an acceptor molecule that is an antibody having a core GlcNAc to which fucose may be attached as an N297-linked glycan or an Fc domain-containing molecule thereof, any reaction method or reaction conditions can be used as long as the reaction proceeds.

[0331] In the present invention, a glycan remodeling antibody or a molecule containing an Fc region thereof can be produced by the method shown in the following scheme, for example, in accordance with the method described in WO2018 / 003983.

[0332] [ka]

[0333] (D-1 process) This step involves hydrolysis of the glycosidic bond between GlcNAcβ1-4GlcNAc in the reducing-end chitobiose structure of the N-linked glycan (N297-linked glycan) attached to asparagine at position 297 of the antibody's amino acid sequence, using a known enzymatic reaction, to produce a glycosylated antibody. The target antibody (1d) (10 mg / mL) is placed in a buffer solution (e.g., phosphate buffer) at 0°C to 40°C, and the glycosidic bond between GlcNAcβ1 and 4GlcNAc in the reducing-end chitobiose structure is hydrolyzed using a hydrolase such as wild-type EndoS enzyme. The reaction time is 10 minutes to 72 hours, preferably 1 hour to 6 hours. The wild-type EndoS enzyme is used in an amount of 0.1 mg to 10 mg, preferably 0.1 mg to 3 mg, per 100 mg of antibody (1d). After the reaction is complete, the antibody is purified using affinity chromatography (HiTrap rProtein A FF (5 ml) (GE Healthcare)) and / or a hydroxyapatite column (Bio-Scale Mini CHT Type I cartridge (5 ml) (BIO-RAD)) to obtain (Fucα1,6)GlcNAc antibody (2d).

[0334] (D-2 process) This step involves conjugating an SG-type glycosylated oxazoline derivative ("[N3-PEG(3)]2-SG(10)-Ox") having an azide group-containing PEG linker to the (Fucα1,6)GlcNAc antibody (2d) obtained in step D-1 using a known enzymatic reaction to produce glycosylated remodeling antibody (3d). The SG-type glycosylated oxazoline derivative ("[N3-PEG(3)]2-SG(10)-Ox") having an azide group-containing PEG linker can be synthesized according to the synthesis method for compound 1-10 described in WO2018 / 003983, as described above.

[0335] Antibody (2d) is reacted with an SG-type sugar chain oxazoline having an azide-containing PEG linker ("[N3-PEG(3)]2-SG(10)-Ox") in a buffer solution (e.g., phosphate buffer) at 0°C to 40°C in the presence of a glycosyltransferase such as EndoS(D233Q / Q303L) to carry out the glycosyltransferase reaction. The reaction time is 10 minutes to 72 hours, preferably 1 to 6 hours. EndoS enzyme (D233Q / Q303L) is used in an amount of 1 mg to 10 mg, preferably 1 mg to 3 mg, per 100 mg of antibody. Two equivalents to an excess of the SG-type sugar chain oxazoline having an azide-containing PEG linker ("[N3-PEG(3)]2-SG(10)-Ox") is used, preferably 4 to 20 equivalents. After the reaction is complete, the product is purified using affinity chromatography (HiTrap rProtein A FF (5 ml) (GE Healthcare)) and a hydroxyapatite column (Bio-Scale Mini CHT Type I cartridge (5 ml) (BIO-RAD)) to obtain the glycan remodeling antibody (3d).

[0336] In preparing the above-mentioned glycan remodeling antibody, concentration of the aqueous antibody solution, concentration measurement, and buffer exchange can be performed in accordance with the general procedures A to C described in WO2020 / 050406.

[0337] As described above, when an azide group-containing modification group is added to the non-reducing end of the "glycan donor molecule" of the present invention, the method for producing a glycan remodeling antibody or an Fc region-containing molecule thereof of the present invention may further include a step of reacting the azide group (N3-) with a molecule having an alkyne structure. This reaction step may be performed either before or after the step of reacting the glycan donor molecule with the acceptor molecule. In the present invention, the "molecule having an alkyne structure" may be any molecule as long as it has an alkyne structure, including, for example, chemotherapeutic agents, molecular targeted drugs, immunostimulators, toxins, antibacterial agents, antiviral agents, diagnostic agents, proteins, peptides, amino acids, nucleic acids, antigens, vitamins, and hormones. In the present invention, the "step of reacting the azide group (N3-) with a molecule having an alkyne structure" can be performed using any reaction method or reaction conditions as long as the reaction proceeds, and can be performed, for example, in accordance with known methods (WO2019 / 065964, WO2020 / 050406, etc.). For example, the "step of reacting the azide group (N3-) with a molecule having an alkyne structure" can be carried out by linking the glycan remodeling antibody (3d) obtained in step D-2 above with a molecule having an alkyne structure via the SPAAC (strain-promoted azide-alkyne cycloaddition: J. Am. Chem. Soc. 2004, 126, 15046-15047) reaction.

[0338] In one aspect, the present invention further provides a method for producing an antibody-drug conjugate, which includes the method for producing the above-mentioned glycan remodeling antibody or its Fc region-containing molecule. In the present invention, the antibody and drug contained in the "antibody-drug conjugate" are not limited as long as they achieve the desired effect, and any substances can be used depending on the purpose. In the present invention, examples of the method for producing an antibody-drug conjugate include, but are not limited to, the above-mentioned steps D-1 and D-2, and a production method comprising the SPAAC reaction.

[0339] In the present invention, the term "antibody-drug conjugate" refers to a complex in which a drug is bound to an antibody via a linker. By using the method of the present invention, a drug can be introduced site-specifically to synthesize a homogeneous antibody-drug conjugate.

[0340] In the present invention, an "antibody drug conjugate" is a compound represented by the following formula (XIII): [ka] It is shown by m 1 is in the range of 1 to 10 and indicates the number of drugs bound per antibody molecule in the antibody-drug conjugate. Ab represents an antibody or a functional fragment of the antibody, L represents a linker connecting Ab and D, and D represents a drug.

[0341] Antibodies used in the present invention include full-body antibodies and functional antibody fragments. A "functional antibody fragment," also known as an "antigen-binding fragment," refers to a partial fragment of an antibody that retains antigen-binding activity, and includes Fab, F(ab'), Fv, scFv, diabody, linear antibody, and multispecific antibody fragments. Antigen-binding fragments of antibodies also include Fab', a monovalent fragment of the antibody variable region obtained by treating F(ab') under reducing conditions. However, they are not limited to these molecules, as long as they retain antigen-binding ability. These antigen-binding fragments also include proteins produced in suitable host cells using genetically engineered antibody genes, as well as full-length antibody protein molecules treated with appropriate enzymes.

[0342] In the present invention, the antibody may be derived from any species, but preferably, examples thereof include human, rat, mouse, and rabbit. When the antibody is derived from a species other than human, it is preferably chimerized or humanized using well-known techniques. Furthermore, in the present invention, the antibody may be a polyclonal antibody or a monoclonal antibody, but monoclonal antibodies are preferred. Monoclonal antibodies include monoclonal antibodies derived from non-human animals such as rat antibodies, mouse antibodies, and rabbit antibodies, chimeric antibodies, humanized antibodies, human antibodies, functional fragments thereof, and modified versions thereof.

[0343] The antibody used in the present invention is not limited as long as it produces the desired effect, and examples thereof include an anti-HER2 antibody, an anti-HER3 antibody, an anti-DLL3 antibody, an anti-FAP antibody, an anti-CDH11 antibody, an anti-CDH6 antibody, an anti-A33 antibody, an anti-CanAg antibody, an anti-CD19 antibody, an anti-CD20 antibody, an anti-CD22 antibody, an anti-CD30 antibody, an anti-CD33 antibody, an anti-CD56 antibody, an anti-CD70 antibody, an anti-CD98 antibody, an anti-TROP2 antibody, an anti-CEA antibody, an anti-Cripto antibody, an anti-EphA2 antibody, an anti-G250 antibody, an anti-MUC1 antibody, an anti-GPNMB antibody, an anti-integrin antibody, an anti-PSMA antibody, an anti-tenascin-C antibody, an anti-SLC44A4 antibody, an anti-Mesothelin antibody, an anti-ENPP3 antibody, an anti-CD47 antibody, an anti-EGFR antibody, an anti-GPR20 antibody, and an anti-DR5 antibody.

[0344] The antibodies used in producing the antibody-drug conjugates of the present invention can be obtained by immunizing an animal with a polypeptide antigen and collecting and purifying the antibodies produced in the body using methods commonly used in this field. The origin of the antigen is not limited to humans; animals can also be immunized with antigens derived from animals other than humans, such as mice or rats. In this case, antibodies applicable to human diseases can be selected by testing the cross-reactivity of the obtained antibodies that bind to heterologous antigens with human antigens.

[0345] Alternatively, a hybridoma can be established by fusing antibody-producing cells that produce antibodies against an antigen with myeloma cells according to known methods (e.g., Kohler and Milstein, Nature (1975) 256, pp. 495-497; Kennett, R. ed., Monoclonal Antibodies, pp. 365-367, Plenum Press, NY (1980)), and a monoclonal antibody can be obtained.

[0346] The antigen can be obtained by genetically manipulating a gene encoding the antigen protein in a host cell to produce it.

[0347] The humanized antibodies used to produce the antibody-drug conjugates of the present invention can be obtained according to known methods (e.g., Proc. Natl. Acad. Sci. USA, 81, 6851-6855, (1984), Nature (1986) 321, p. 522-525, WO90 / 07861).

[0348] For example, anti-HER2 antibodies (US5821337, WO2004 / 008099, WO2020 / 050406, etc.), anti-CD33 antibodies (WO2014 / 057687, WO2020 / 050406, etc.), anti-EphA2 antibodies (WO2009 / 028639, WO2020 / 050406, etc.), anti-CDH6 antibodies (WO2018 / 028639, WO2020 / 050406, etc.), Antibodies such as WO2019 / 050433, WO2002 / 092771, and the like can be obtained by known means.

[0349] Drugs used in the present invention are not limited as long as they produce the desired effect, and include pharmaceutically active compounds such as chemotherapeutic agents, molecular targeted drugs, immune activators, toxins, antibacterial agents, antiviral agents, diagnostic agents, proteins, peptides, amino acids, nucleic acids, antigens, vitamins, and hormones.

[0350] Drug D may be, for example, a compound of the formula: [ka] It is shown as TIFF2025160360000287.tif216161 (wherein the asterisk indicates binding to the linker L).

[0351] In the present invention, the linker L that connects the antibody Ab and the drug D has the following formula: -Lb-La-Lp-Lc-* (where the asterisk indicates binding to drug D).

[0352] Lp may or may not have a linker consisting of an amino acid sequence that is cleavable in target cells, and specific examples thereof include -GGFG-, -GGPI-, -GGVA-, -GGFM-, -GGVCit-, -GGFCit-, -GGICit-, -GGPL-, -GGAQ-, or -GGPP-.

[0353] La is -C(=O)-(CH2CH2)n 2 -C(=O)-, -C(=O)-(CH2CH2)n 2 -CH2-C(=O)-, -C(=O)-(CH2CH2)n 2 -C(=O)-NH-(CH2CH2)n 3 -C(=O)-, -C(=O)-(CH2CH2)n 2 -C(=O)-NH-(CH2CH2)n 3 -CH2-C(=O)-, -C(=O)-(CH2CH2)n 2 -C(=O)-NH-(CH2CH2O)n 3 -CH2-C(=O)-, -(CH2)n 4 -OC(=O)-, or -(CH2)n 9 -C(=O)- (where n 2represents an integer from 1 to 3, and n 3 represents an integer from 1 to 5, and n 4 represents an integer from 0 to 2, and n 9 represents an integer of 2 to 7).

[0354] Lb represents a spacer that connects the sugar chain or remodeled sugar chain of La and Ab, and specific examples thereof include: [ka] or [ka] (In the structural formula of Lb shown above, the asterisk indicates binding to La, and the wavy line indicates binding to the sugar chain of Ab or a remodeled sugar chain.)

[0355] Lc is -NH-CH2-, -NH-B-CH2-O(C=O)-, or is absent, where B is a 1,4-phenyl group, a 2,5-pyridyl group, a 3,6-pyridyl group, a 2,5-pyrimidyl group, or a 2,5-thienyl group.

[0356] The present invention will be described below with reference to examples, but is not limited thereto. In the following, "compound I-1" or "I-1" in the synthesis scheme indicates that it corresponds to "a compound represented by formula I-1," and the same applies to the subsequent compound numbers. [Example]

[0357] In the following examples, room temperature refers to 15°C to 35°C. Silica gel chromatography was performed using a Biotage Sfar HC D (20 μm, manufactured by Biotage). Reverse-phase column chromatography was performed using a Universal Column ODS Premium 30 μm L size (manufactured by Yamazen Corporation) and an Inject column ODS L size (manufactured by Yamazen Corporation). Preparative HPLC was performed using an Agilent Preparative HPLC System (manufactured by Agilent Technology). The preparative column used was an XBridge Prep OBD (5 μm, C18, 130 Å, 250 × 30 mm, manufactured by Waters).

[0358] The following instruments were used to measure various spectral data. 1 H-NMR and 13 C-NMR spectra were measured using JEOL ECZ500R and ECX400P instruments. Mass spectra were measured using Shimadzu LCMS-2010 and LCMS-2020 (Shimadzu Corporation), XEVO Q-Tof MS (Waters), and Q-Exactive (Thermo Fisher).

[0359] <Synthesis of Compound III-13> Compound III-13 was synthesized according to the following synthetic scheme 1A. [Synthetic Scheme 1A]

[0360] [ka]

[0361] Example 1: 1,2:5,6-bis-O-(1-methylethylidene)-3-O-(2-naphthalenylmethyl)-α-D-glucofuranose (Compound I-2)

[0362] [ka]

[0363] (Small process 1-1) A solution of sodium hydride (55.32 g, 1.38 mol, content: 50-72%) in tetrahydrofuran (900 mL) was cooled to 0 °C, and a solution of 1,2:5,6-bis-O-(1-methylethylidene)-α-D-glucofuranose (Compound I-1) (300.00 g, 1.15 mol) in tetrahydrofuran (1.05 L) was added dropwise over 1 hour. The mixture was then heated to 25 °C, and 1,3-dimethyl-2-imidazolidinone (150 mL) and 2-bromomethylnaphthalene (280.31 g, 1.27 mol) were added. After stirring at 25 °C for 6 hours, the completion of the reaction was confirmed by HPLC. Ethylenediamine (anhydrous) (13.85 g, 230.52 mmol) was added, and the mixture was stirred for an additional 1 hour. The solution was cooled to 0°C, and 10% aqueous citric acid (1.2 L) was added over 1 hour. The reaction mixture was diluted with heptane (3 L) and separated into an organic layer and an aqueous layer. The organic layer was washed with water (900 mL) and then concentrated under reduced pressure to a volume of 900 mL. Acetonitrile (3 L) was added, and the mixture was again concentrated to a volume of 900 mL to obtain crude 1,2:5,6-bis-O-(1-methylethylidene)-3-O-(2-naphthalenylmethyl)-α-D-glucofuranose (Compound I-2) as an acetonitrile solution. This product was used directly in the next step.

[0364] Example 2: 3-O-(2-naphthalenylmethyl)-D-glucopyranose (Compound I-3)

[0365] [ka]

[0366] (Small process 1-2) Acetonitrile (1.5 L), water (600 mL), and concentrated hydrochloric acid (17.51 ​​g, 172.89 mmol) were added to the solution of crude compound I-2 obtained in Example 1, and the mixture was stirred at 55°C for 18.5 hours. After confirming the completion of the reaction by HPLC, the reaction solution was cooled to 0°C, and the pH of the system was adjusted to 6.25 with 4 N aqueous sodium hydroxide solution (43.22 mL). The reaction solution was diluted with heptane (900 mL), and the mixture was separated into an acetonitrile layer and a heptane layer. Ethyl acetate (2.4 L) and water (600 mL) were added to the acetonitrile layer, and the mixture was separated to obtain organic layer 1 and an aqueous layer. A mixed solution of ethyl acetate (1.5 L) and tetrahydrofuran (1.5 L) was added to the aqueous layer again, and the mixture was separated to obtain organic layer 2 and an aqueous layer. The organic layers 1 and 2 were mixed, washed with saturated brine (600 mL), and concentrated under reduced pressure to a volume of 1.5 L (crystal precipitation was confirmed during the concentration step). Ethyl acetate (4.5 L) was added, and the mixture was concentrated again to a volume of 3 L. Ethyl acetate (1.5 L) and cyclopentyl methyl ether (1.5 L) were added to the suspension, and the mixture was stirred at 55 °C for 1 hour. Heptane (3 L) was added dropwise over 1.5 hours, stirred for 1 hour, and then cooled to 0 °C. The precipitated crystals were then filtered and washed with a mixture of ethyl acetate (1.2 L) and heptane (600 mL) cooled to 0 °C. The resulting crystals were dried under reduced pressure at 40 °C to obtain 3-O-(2-naphthalenylmethyl)-D-glucopyranose (Compound I-3) (356.95 g, yield 96.7%, HPLC area: 98.47%).

[0367] 1 H-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),4.94-5.0 0(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 - )[MH] - calcd for C 17 H 19 O6:319.1187;found 319.1175.

[0368] Example 3: 2,4,6-Tri-O-acetyl-3-O-(2-naphthalenylmethyl)-D-glucopyranose (Compound I-5)

[0369] [ka]

[0370] (Small process 1-3&1-4) To a solution of 3-O-(2-naphthalenylmethyl)-D-glucopyranose (Compound I-3) (150.00 g, 468.25 mmol) in tetrahydrofuran (675 mL), triethylamine (236.92 g, 2.34 mol) and 4-dimethylaminopyridine (0.29 g, 2.34 mmol) were added and cooled to 0 °C. Then, acetic anhydride (195.99 g, 1.92 mol) was added dropwise over 30 minutes. The mixture was then heated to 25 °C and stirred for 3 hours. HPLC analysis confirmed the formation of 1,2,4,6-tetra-O-acetyl-3-O-(2-naphthalenylmethyl)-D-glucopyranose (Compound I-4). The reaction mixture was cooled to 10 °C, and 1-methylpiperazine (60.97 g, 608.73 mmol) was added. After stirring at 35°C for 18 hours, the completion of the reaction was confirmed by HPLC and the mixture was cooled to 0°C. The pH was adjusted to 6.36 with 6N aqueous hydrochloric acid (480 mL) and then diluted with heptane (375 mL). The organic and aqueous layers were separated. The organic layer was washed with saturated aqueous sodium bicarbonate (450 mL) and water (450 mL) and then concentrated under reduced pressure to 450 mL. Ethyl acetate (2.25 L) was added, and the mixture was concentrated again to 450 mL. The same procedure was repeated once more. Dichloromethane (2.25 L) was added to this solution, and the mixture was concentrated to 450 mL. The same procedure was repeated once more to obtain crude 2,4,6-tri-O-acetyl-3-O-(2-naphthalenylmethyl)-D-glucopyranose (Compound I-5) as a dichloromethane solution. This product was used directly in the next step.

[0371] Example 4: 2,4,6-Tri-O-acetyl-3-O-[(naphthalen-2-yl)methyl]-1-O-(2,2,2-trichloroethanimidoyl)-D-glycero-hexopyranose (Compound I-6)

[0372] [ka]

[0373] (Small process 1-5) To a solution of crude 2,4,6-tri-O-acetyl-3-O-(2-naphthalenylmethyl)-D-glucopyranose (Compound I-5) obtained in Example 3, dichloromethane (450 mL) and trichloroacetonitrile (338.03 g, 2.34 mol) were added and cooled to 0 °C. 1,8-diazabicyclo[5.4.0]-7-undecene (5.70 g, 37.46 mmol) was then added dropwise. After stirring at 0 °C for 14.5 hours, the completion of the reaction was confirmed by HPLC, and acetic acid (2.25 g, 37.46 mmol) was added. Silica gel 60N (spherical, neutral) (150 g) was added to the solution, followed by stirring for 1.5 hours and filtration. The silica gel was washed with dichloromethane (1.5 L), and the filtrate was concentrated under reduced pressure to a volume of 450 mL. Further dichloromethane (1.5 L) was added and concentrated to a volume of 450 mL to give crude 2,4,6-tri-O-acetyl-3-O-[(naphthalen-2-yl)methyl]-1-O-(2,2,2-trichloroethanimidoyl)-D-glycerohexopyranose (Compound I-6) as a dichloromethane solution, which was used directly in the next step.

[0374] Example 5: 4-Methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-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 III-1)

[0375] [ka]

[0376] (Small process 1-6) To a solution of crude 2,4,6-tri-O-acetyl-3-O-[(naphthalen-2-yl)methyl]-1-O-(2,2,2-trichloroethanimidoyl)-D-glycerohexopyranose (Compound I-6) obtained in Example 4, 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound II-1) (276.66 g, 468.25 mmol), dichloromethane (4.2 L), and molecular sieves 4A (83.00 g) were added and cooled to -5 ° C. Trimethylsilyl trifluoromethanesulfonate (10.41 g, 46.83 mmol) was added dropwise to this suspension over 20 minutes, followed by stirring for 3 hours. After confirming the completion of the reaction by HPLC, triethylamine (23.69 g, 234.13 mmol) was added. The suspension was filtered and washed with ethyl acetate (2.8 L). The filtrate was concentrated under reduced pressure to a volume of 1.4 L. Ethyl acetate (4.2 L) was added and concentrated to a volume of 1.4 L. The same procedure was repeated once more. Ethyl acetate (2.8 L) was added to this solution, and the mixture was washed with saturated aqueous sodium bicarbonate (830 mL) and water (830 mL). The organic layer was concentrated under reduced pressure to a volume of 830 mL. 2-Propanol (4.2 L) was added and concentrated to a volume of 1.4 L. The suspension was then heated to 65°C. Ethyl acetate (830 mL) was added and stirred at 65°C for 2 hours, after which 2-propanol (5.53 L) was added dropwise over 2 hours. After cooling this suspension to 0° C., the crystals were filtered and washed with 2-propanol (1.4 L) cooled to 0° C. The obtained crystals were dried under reduced pressure at 40° C. to give crude 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-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 III-1) (355.34 g, yield 90.5%, HPLC area: 94.46%).

[0377] The crude compound III-1 (350.00 g) was dissolved in methyl isobutyl ketone (2.1 L) at 50° C., and then ethylcyclohexane (1.4 L) was added dropwise over 1 hour. Seed crystals (70.00 mg) of 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-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 III-1) were added and stirred for 1 hour. After confirming the precipitation of crystals, ethylcyclohexane (4.9 L) was added dropwise over 2 hours. The suspension was cooled to room temperature and stirred for 14.5 hours. The precipitated crystals were filtered and washed with a mixture of methyl isobutyl ketone (350 mL) and ethylcyclohexane (1.4 L). The resulting crystals were dried under reduced pressure at 40 °C to give 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-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 III-1) (331.74 g, yield 94.8%, HPLC area: 98.07%).

[0378] 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). 13 C-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] + calcd for C 58 H 58 NO 16 :1024.3750;found 1024.3706.

[0379] The spectra of the obtained compound were confirmed to match those in the following literature: Reference 1) Org. Biomol. Chem., 2018, 16, 4720-4727.

[0380] Example 6: 4-Methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-4-O-{3-O-[(naphthalen-2-yl)methyl]-β-D-glucopyranosyl}-β-D-glucopyranoside (Compound III-2)

[0381] [ka]

[0382] (Small process 1-7) To a solution of 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-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 III-1) (30.00 g, 29.29 mmol) in tetrahydrofuran (150 mL), methanol (90 mL) and methyl trifluoroacetate (3.75 g, 29.29 mmol) were added and stirred at 25 ° C for 10 minutes. Potassium tert-butoxide (1 mol / L tetrahydrofuran solution) (14.7 mL, 14.65 mmol) was then added. The mixture was then heated to 55 ° C and stirred for 2 hours. The completion of the reaction was confirmed by HPLC. The reaction mixture was cooled to 25 °C, and acetic acid (1.76 g, 29.29 mmol) and ethyl acetate (300 mL) were added, in that order. This solution was washed twice with 1% brine (300 mL) and then concentrated under reduced pressure to 90 mL. Ethyl acetate (450 mL) was added, and the mixture was concentrated again to 90 mL. Acetonitrile (450 mL) was added, and the mixture was concentrated to 90 mL to obtain crude 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-4-O-{3-O-[(naphthalen-2-yl)methyl]-β-D-glucopyranosyl}-β-D-glucopyranoside (Compound III-2) as an acetonitrile solution. This product was used directly in the next step.

[0383] Example 7: 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-3)

[0384] [ka]

[0385] (Small process 1-8) Acetonitrile (210 mL), benzaldehyde dimethyl acetal (5.13 g, 33.69 mmol), and p-toluenesulfonic acid monohydrate (0.17 g, 0.88 mmol) were added to a solution of crude 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-4-O-{3-O-[(naphthalen-2-yl)methyl]-β-D-glucopyranosyl}-β-D-glucopyranoside (Compound III-2) obtained in Example 6, and the mixture was stirred for 30 minutes at 25 ° C. Toluene (600 mL) was added to the solution, which was then concentrated to a volume of 300 mL. At this point, completion of the reaction was confirmed by HPLC. Further 1-methylimidazole (12.03 g, 146.47 mmol) was added and the mixture was concentrated to 90 mL to give crude 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-3) as a toluene solution containing 1-methylimidazole, which was used directly in the next step.

[0386] Example 8: 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 (Compound III-4, X1 = trifluoromethanesulfonyl group)

[0387] [ka]

[0388] (Small process 1-9) To a solution of crude 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-3) obtained in Example 7 (containing 1-methylimidazole), ethyl acetate (210 mL) was added and cooled to 0 °C. Trifluoromethanesulfonic anhydride (16.53 g, 58.59 mmol) was added dropwise over 1 hour, followed by stirring for 30 minutes. After confirming the completion of the reaction 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) and once with saturated brine (150 mL), then concentrated under reduced pressure to 90 mL. Ethyl acetate (300 mL) was added, and the mixture was concentrated again to 90 mL to give crude 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 (Compound III-4, X1 = trifluoromethanesulfonyl group) as an ethyl acetate solution. This product was used directly in the next step.

[0389] Example 9: 4-Methoxyphenyl 3,6-di-O-benzyl-4-O-{4,6-O-benzylidene-3-O-[(naphthalen-2-yl)methyl]-β-D-mannopyranosyl}-2-(2-carboxybenzamido)-2-deoxy-β-D-glucopyranoside (Compound III-6)

[0390] [ka]

[0391] (Small process 1-10&1-11) Dimethyl sulfoxide (150 mL) and tetrabutylammonium acetate (17.67 g, 58.59 mmol) were added to a solution of crude 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 (Compound III-4, X1 = trifluoromethanesulfonyl group) obtained in Example 8. The mixture was heated to 30° C. and stirred for 17 hours. After stirring, the 4-methoxyphenyl The formation of 4-O-{2-O-acetyl-4,6-O-benzylidene-3-O-[(naphthalen-2-yl)methyl]-β-D-mannopyranosyl}-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-5, X2 = acetyl group) was confirmed. Toluene (150 mL) was added to the reaction mixture, and the mixture was concentrated under reduced pressure to a volume of 165 mL. Methanol (45 mL) and 50% aqueous sodium hydroxide solution (3.52 g, 87.88 mmol) were added, and the mixture was stirred at 25 °C for 1.5 hours. After confirming the completion of the reaction by HPLC, ethyl acetate (450 mL) and water (300 mL) were added and the mixture was separated. Water (300 mL) was added to the organic layer, cooled to 0°C, and the pH was adjusted to 2.73 with 6 N hydrochloric acid under vigorous stirring. Tetrahydrofuran (300 mL) was added to the separated organic layer, and the mixture was concentrated under reduced pressure to a volume of 150 mL. Tetrahydrofuran (300 mL) was added, and the mixture was concentrated again to a volume of 90 mL, and the internal temperature was adjusted to 45°C. After adding tetrahydrofuran (60 mL), the mixture was cooled to 25°C, and 2-propanol (150 mL) and water (15 mL) were added. Seed crystals 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-carboxybenzamido)-2-deoxy-β-D-glucopyranoside (Compound III-6) (30 mg) were added.The mixture was stirred at 25°C for 14 hours, and after confirming the precipitation of crystals, 2-propanol (210 mL) was added dropwise over 1 hour, followed by cooling to 0°C. After stirring for 2 hours, the precipitated crystals were filtered and washed with 2-propanol (150 mL) cooled to 0°C. The resulting crystals were dried under reduced pressure at 40°C 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-carboxybenzamido)-2-deoxy-β-D-glucopyranoside (Compound III-6) (27.74 g, yield 94.3%, HPLC area: 97.85%). Compound III-4 (X1 = trifluoromethanesulfonyl group) can also be converted to compound III-5 (X2 = acetyl group) using cesium acetate (3 equivalents), dimethyl sulfoxide, and 50°C for 24 hours. Compound III-6 can then be obtained by the same reaction (compound III-5 (X2 = acetyl group) → compound III-6) and post-treatment.

[0392] 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). 13 C-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] + calcd for C 59 H 58 NO 14 :1004.3852;found 1004.3873.

[0393] Example 10: 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-8)

[0394] [ka]

[0395] (Small process 1-12) 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-carboxybenzamido)-2-deoxy-β-D-glucopyranoside (Compound III-6) (3.00 g, 2.99 mmol) in dichloromethane (30 mL), 1-hydroxybenzotriazole monohydrate (91.8 mg, 0.60 mmol), N,N-diisopropylethylamine (0.42 g, 3.29 mmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.63 g, 3.29 mmol) were added. The mixture was heated to 40 °C and stirred for 24 hours. After confirming the completion of the reaction by HPLC, the reaction solution was cooled to 25 °C, separated with water (30 mL), and the organic layer was washed with saturated brine (15 mL). The solution was concentrated to dryness under reduced pressure to give crude 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-8). This product was used directly in the next step.

[0396] Example 11: 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-9)

[0397] [ka]

[0398] (Small process 1-15) The crude 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-8) obtained in Example 10 was added with N,N-dimethylacetamide (30 mL), benzyl bromide (0.77 g, 4.48 mmol), and molecular sieves 4A (900 mg), and the mixture was cooled to 0°C. Sodium hydride (0.16 g, 3.88 mmol, content: 50-72%) was added and stirred for 3 hours. After confirming the completion of the reaction by HPLC, the reaction solution was cooled to 25°C, and ethylenediamine (anhydrous) (0.18 g, 2.99 mmol) was added and stirred for an additional 1 hour. This solution was cooled to 0°C, and acetic acid (0.36 g, 5.98 mmol) was added and filtered. After washing the molecular sieves 4A with N,N-dimethylacetamide (15 mL), ethyl acetate (60 mL) and water (30 mL) were added and the mixture was separated. The organic layer was washed twice with water (30 mL) and once with saturated brine (15 mL). This solution was concentrated to dryness under reduced pressure, and toluene (30 mL) and silica gel 60N (spherical) (3 g) were added and stirred at 25°C for 30 minutes. The suspension was filtered, and the silica gel was washed with a mixed solution of toluene (82 mL) and ethyl acetate (8 mL). The filtrate was concentrated to dryness under reduced pressure to give crude 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-9).

[0399] Example 12: 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-10)

[0400] [ka]

[0401] (Small process 1-16) The crude 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-9) obtained in Example 11 was added with dichloromethane (30 mL) and molecular sieves 4A (300 mg) and cooled to 0 °C. Borane-tetrahydrofuran complex (0.91 mol / L tetrahydrofuran solution) (16.42 mL, 14.94 mmol) and copper(II) trifluoromethanesulfonate (0.16 g, 0.45 mmol) were added and stirred for 6 hours. After confirming the completion of the reaction by HPLC, methanol (3 mL) was added and the mixture was stirred for an additional 1 hour. The solution was filtered, and the molecular sieves 4A were washed with ethyl acetate (30 mL). The organic layer was then washed twice with 0.5 N hydrochloric acid (30 mL) and once with saturated brine (15 mL). The solution was concentrated under reduced pressure to a volume of 6 mL. Toluene (15 mL) was added, and the mixture was again concentrated to a volume of 6 mL to obtain a toluene solution of crude 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-10).

[0402] Example 13: 4-Methoxyphenyl 3,6-di-O-benzyl-2-deoxy-4-O-{2,4-di-O-benzyl-β-D-mannopyranosyl}-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-11)

[0403] [ka]

[0404] (Small process 1-17) A solution of crude 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-10) was added to dichloromethane (30 mL), water (1.5 mL), and potassium dihydrogen phosphate (0.81 g, 5.98 mmol). The mixture was cooled to 0 ° C., and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (0.81 g, 3.59 mmol) was added. The mixture was heated to 25 ° C. and stirred for 8 hours. The completion of the reaction was confirmed by HPLC. The reaction solution was cooled to 0 °C, and ethyl acetate (60 mL) and 5% aqueous sodium sulfite solution (30 mL) were added and the mixture was separated. The organic layer was washed twice with 5% aqueous sodium sulfite solution (30 mL) and once with saturated brine (15 mL), and then concentrated to dryness under reduced pressure. This product was purified by silica gel column chromatography (silica gel 300 g, hexane:ethyl acetate = 50:50 → 30:70) to give 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-4-O-{2,4-di-O-benzyl-β-D-mannopyranosyl}-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-11) (2.15 g, yield 76.7%, HPLC area: 98.67%).

[0405] 1H-NMR(500MHz,CDCl3)δ7.67(br,4H),7.27-7.37(m,15H),6.89-6.96(m,5H),6.83(ddd,J=9.5,4.0,2.5Hz,2H),6.70(ddd,J=9.0,4.0,2.5Hz,2H),5.64(d,J=9.0Hz,1H),5.02(d,J=11.5Hz,1H),4.94(d,J=12.0Hz,1H),4.82(d,J=11.5Hz,1H),4.70(d,J=12.0Hz,1H),4.66(d,J=12.0Hz,1H),4.62(s,1H),4.57(d,J=11.0Hz,1H),4.51(d,J=12.0Hz,1H),4.42-4.46(m,2H),4.33(dd,J=10.5,8.5Hz,1H),4.09(t,J=9.0Hz,1H),3.69-3.79(m,4H),3.71(s,3H),3.65(d,J=3.5Hz,1H),3.51(td,J=9.0,3.5Hz,1H),3.45(t,J=9.0Hz,1H),3.44-3.48(m,1H),3.17(m,1H),2.29(d,J=9.0Hz,1H),1.92(t,J=6.0Hz,1H). 13 C-NMR (125MHz,CDCl3)δ155.6,151.0,138.54,138.46,138.3,137.8,134.1,131.8,128.8,128.6,128.3,128.21,128.19,128.15,128.1,128.0,127.5,127.4,123.6,118.9,114.6,101.5,97.9,79.1,78.5,77.3,76.7,75.6,75.4,75.3,74.9,74.8,74.4,73.9,68.5,62.5,55.8. HRMS(ESI - )[M-H] - calcd for C 55 H 54 NO 13 :936.3601;found 936.3592.

[0406] The spectra of the obtained compound were confirmed to match those in the following literature: Reference 1) Org. Biomol. Chem., 2018, 16, 4720-4727.

[0407] Example 14: 4-Methoxyphenyl 2-amino-3,6-di-O-benzyl-2-deoxy-4-O-(2,4-di-O-benzyl-β-D-mannopyranosyl)-β-D-glucopyranoside (Compound III-12)

[0408] [ka]

[0409] (Small process 1-18) A solution of 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-4-O-{2,4-di-O-benzyl-β-D-mannopyranosyl}-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-11) (700 mg, 0.75 mmol) in 2-methyltetrahydrofuran (3.5 mL) and n-butanol (3.5 mL) was added with anhydrous ethylenediamine (0.25 mL, 3.73 mmol), and the mixture was heated to 80 °C and stirred for 16.5 hours. After confirming the completion of the reaction by HPLC, the mixture was cooled to room temperature, and cyclopentyl methyl ether (7 mL) and 10% aqueous methanol (2.8 mL) were added. The aqueous layer was separated, and the resulting organic layer was washed twice with 10% aqueous methanol (2.8 mL) and then concentrated to dryness under reduced pressure to give crude 4-methoxyphenyl 2-amino-3,6-di-O-benzyl-2-deoxy-4-O-(2,4-di-O-benzyl-β-D-mannopyranosyl)-β-D-glucopyranoside (Compound III-12).

[0410] Example 15: 4-Methoxyphenyl 3,6-di-O-benzyl-2-deoxy-4-O-(2,4-di-O-benzyl-β-D-mannopyranosyl) 2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranoside (Compound III-13)

[0411] [ka]

[0412] (Small process 1-19) The crude 4-methoxyphenyl 2-amino-3,6-di-O-benzyl-2-deoxy-4-O-(2,4-di-O-benzyl-β-D-mannopyranosyl)-β-D-glucopyranoside (Compound III-12) was added to tetrahydrofuran (5.6 mL) and water (0.7 mL), followed by sodium bicarbonate (125.4 mg, 1.49 mmol) and 2,2,2-trichloroethyl chloroformate (189.7 mg, 0.90 mmol). After stirring at 25 °C for 2 hours, the resulting suspension was added with water (2.8 mL) and cyclopentyl methyl ether (7 mL). After removing the aqueous layer, the crude product was concentrated to dryness under reduced pressure and purified by silica gel column chromatography (ethyl acetate / hexane 20% → 65%) to give 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-4-O-(2,4-di-O-benzyl-β-D-mannopyranosyl) 2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranoside (Compound III-13) (702.6 mg, yield 96%).

[0413] 1H-NMR(500MHz,CDCl3)δ1.94(br,1H),2.34(d,J=9.0Hz,1H),3.14(m,1H),3.40-3.60(m,4H),3.60-3.80(m,5H), 3.74(s,3H),3.98(t,J=9.0Hz,1H),4.02(br,1H),4.45(d,J=7.0Hz,1H),4.46(brs,1H),4.58(d,J=11.5Hz,1H), 4.59-4.65(m,4H),4.68(d,J=12.0Hz,1H),4.82(d,J=11.0Hz,1H),4.93(d,J=11.0Hz,1H),5.01(d,J=10.5Hz,1H ),5.23(d,J=5.5Hz,1H),5.54(d,J=7.0Hz,1H),6.77(d,J=9.0Hz,2H),6.96(d,J=9.0Hz,2H),7.22-7.38(m,20H). 13 C-NMR(125MHz,CDCl3)δ55.4, 57.2, 62.1, 68.5, 73.6, 74.0, 74.2, 74.3, 74.6, 74.9, 75.1, 75.3, 76.4, 77.8, 77.9, 78.2, 95.4, 99.4, 101.4, 114.4, 118.5, 127.4, 127.65, 127.75, 127.82, 127.9, 128.3, 128.4, 128.46, 128.50, 137.5, 138.0, 138.2, 128.3, 151.2, 153.9, 155.3. MS(ESI)(m / z):999([M+NH4] + ). =999

[0414] <Synthesis of Compound III-8> Compound III-8 was synthesized according to the following synthetic scheme 1B. [Synthetic Scheme 1B]

[0415] [ka]

[0416] Example 16: 4-Methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-4-O-{3-O-[(naphthalen-2-yl)methyl]-β-D-glucopyranosyl}-β-D-glucopyranoside (Compound III-2)

[0417] [ka]

[0418] (Small process 1-7) To a solution of 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-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 III-1) (50.00 g, 48.82 mmol) in tetrahydrofuran (250 mL), methanol (75 mL) and methyl trifluoroacetate (6.25 g, 48.82 mmol) were added and stirred at 25 ° C for 30 minutes, followed by the addition of potassium tert-butoxide (1 mol / L tetrahydrofuran solution) (24.41 mL, 24.41 mmol). The mixture was then heated to 55 ° C and stirred for 3.5 hours, after which completion of the reaction was confirmed by HPLC. The reaction mixture was cooled to 25 °C, and acetic acid (2.93 g, 48.82 mmol) was added. The mixture was concentrated under reduced pressure to 150 mL. Ethyl acetate (500 mL) and water (150 mL) were then added and the mixture was separated. The organic layer was concentrated under reduced pressure to 150 mL. Acetonitrile (750 mL) was added, and the mixture was concentrated again to 150 mL. Acetonitrile (500 mL) was added, and the mixture was concentrated to 250 mL. Crude 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-4-O-{3-O-[(naphthalen-2-yl)methyl]-β-D-glucopyranosyl}-β-D-glucopyranoside (Compound III-2) was obtained as an acetonitrile solution. This product was used directly in the next step.

[0419] Example 17: 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-3)

[0420] [ka]

[0421] (Small process 1-8) Acetonitrile (250 mL), benzaldehyde dimethyl acetal (14.86 g, 97.65 mmol), and p-toluenesulfonic acid monohydrate (0.46 g, 2.44 mmol) were added to a solution of crude 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-4-O-{3-O-[(naphthalen-2-yl)methyl]-β-D-glucopyranosyl}-β-D-glucopyranoside (Compound III-2) obtained in Example 16, and the mixture was stirred for 30 minutes at 25° C. The reaction solution was concentrated to a volume of 150 mL, at which point completion of the reaction was confirmed by HPLC. To this solution, triethylamine (1.48 g, 14.65 mmol), ethyl acetate (500 mL), and saturated aqueous sodium bicarbonate (150 mL) were added, and the layers were separated. The organic layer was washed with 2 N hydrochloric acid (150 mL), and the separated organic layer was concentrated under reduced pressure to a volume of 150 mL. Ethyl acetate (500 mL) was added, and the mixture was concentrated again to a volume of 250 mL to obtain crude 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-3) as an ethyl acetate solution. This product was used directly in the next step.

[0422] Example 18: 4-Methoxyphenyl 3,6-di-O-benzyl-4-O-{4,6-O-benzylidene-3-O-[(naphthalen-2-yl)methyl]hexopyranosyl-2-ulose}-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-7)

[0423] [ka]

[0424] (Small process 1-13) To a solution of crude 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-3), dichloromethane (150 mL) and iodobenzene diacetate (18.87 g, 58.59 mmol) were added and cooled to 0 ° C. 9-Azanoradamantane N-oxyl (0.67 g, 4.88 mmol) was added to this solution, and the mixture was heated to 25 ° C. and stirred for 5 hours. After confirming the completion of the reaction by HPLC, the reaction mixture was cooled to 0°C, and ethyl acetate (600 mL) and 10% aqueous sodium sulfite solution (250 mL) were added, followed by separation. The organic layer was washed with 2N hydrochloric acid (150 mL) and saturated brine (100 mL) and concentrated under reduced pressure to a volume of 150 mL. The internal temperature was adjusted to 50°C, and 2-propanol (1 L) was added dropwise over 1 hour. The suspension was then cooled to 0°C and stirred for 3 hours. The precipitated crystals were then filtered and washed with a mixed solvent of 2-propanol (200 mL) and ethyl acetate (50 mL) cooled to 0°C. The resulting crystals were dried under reduced pressure at 50°C to give 4-methoxyphenyl 3,6-di-O-benzyl-4-O-{4,6-O-benzylidene-3-O-[(naphthalen-2-yl)methyl]hexopyranosyl-2-ulose}-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-7) (41.61 g, yield 86.6%, HPLC area: 99.20%).

[0425] 1H-NMR(500MHz,CDCl3)δ7.86(br,1H),7.80-7.83(m,3H),7.65-7.67(m,4H),7.40-7.52(m,8H),7.18-7.23(m,4H),7.10-7.14(m,1H),7.00-7.02(m,2H),6.89-6.92(m,2H),6.83-6.87(m,1H),6.80(ddd,J=9.0,4.0,2.0Hz,2H),6.70(ddd,J=9.0,4.0,2.5Hz,2H),5.63(d,J=8.5Hz,1H),5.49(s,1H),5.06(d,J=13.0Hz,1H),4.95(d,J=0.5Hz,1H),4.85(d,J=12.5Hz,1H),4.76(d,J=12.5Hz,1H),4.68(d,J=12.0Hz,1H),4.47(dd,J=10.5,8.5Hz,1H),4.40-4.42(m,1H),4.39(t,J=4.5Hz,1H),4.31(d,J=12.0Hz,1H),4.25(dd,J=10.5,5.0Hz,1H),4.17(t,J=9.0Hz,1H),3.99(d,J=11.0Hz,1H),3.89(dd,J=11.5,3.5Hz,1H),3.78-3.86(m,3H),3.71(s,3H),3.57(t,J=10.0Hz,1H),3.45(td,J=10.0,5.0Hz,1H). 13 C-NMR (125MHz,CDCl3)δ197.2,155.7,151.0,138.4,138.0,137.1,135.0,134.0,133.5,133.3131.8,129.4,128.7,128.5,128.4,128.20,128.18,128.1,128.0,127.90,127.87,127.4,126.7,126.4,126.3,126.2,125.8,123.6,119.0,114.6,102.2,101.4,98.0,82.28,82.25,80.8,77.3,75.2,74.7,73.6,73.4,68.7,68.3,66.3,56.0,55.8. HRMS(ESI - )[M+HCO2] - calcd for C 60 H 54 NO15 :1028.3499;found 1028.3531.

[0426] Example 19: 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-8)

[0427] [ka]

[0428] (Step 1-14) (Condition 1: L-Selectride) A solution of compound III-7 (1.50 g, 1.52 mmol) in tetrahydrofuran (22.5 mL) was cooled to -50 °C. After stirring for 50 min, a solution of L-selectride in tetrahydrofuran (1.0 M, 1.68 mL, 1.68 mmol) was added over 5 min. After stirring for 1 h, acetic acid (0.18 mL, 3.05 mmol) was added and the mixture was warmed to room temperature (compound III-7: 2.41% PA, compound III-8: 76% PA @ 220 nm, dr = 96.5:3.5). Ethyl acetate (15 mL) was added to the resulting reaction solution, followed by 5% aqueous citric acid (6 mL) and stirring for 10 min. Heptane (4 mL) was added, the aqueous layer was removed, and the resulting organic layer was washed with water (3 mL). The obtained organic layer was concentrated to dryness and then purified by silica gel chromatography (ethyl acetate / hexane 20%→55%) to obtain compound III-8 (1.15 g, yield 77%).

[0429] (Condition 2: LS-Selectride) A solution of 4-methoxyphenyl 3,6-di-O-benzyl-4-O-{4,6-O-benzylidene-3-O-[(naphthalen-2-yl)methyl]hexopyranosyl-2-ulose}-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-7) (5.00 g, 5.08 mmol) in tetrahydrofuran (50 mL) was added with molecular sieves 4A (500 mg) and cooled to −45° C. After stirring for 15 min, a solution of LS-selectride in tetrahydrofuran (1.0 M, 5.59 mL, 5.59 mmol) was added dropwise over 20 min. After stirring for 2 hours, the completion of the reaction was confirmed by HPLC. Acetic acid (0.38 mL, 6.61 mmol) was added, and the mixture was heated to 25 °C (Compound III-8: 87% PA @ 220 nm, dr = 98.1:1.9). The mixture was filtered through molecular sieves 4A and washed with tetrahydrofuran (10 mL). Ethyl acetate (60 mL) was added to the resulting solution, followed by 5% aqueous citric acid solution (20 mL) and stirring for 10 minutes. After removing the aqueous layer, the resulting organic layer was washed with water (10 mL), 5% aqueous sodium bicarbonate solution (20 mL), and water (15 mL). The organic layer was concentrated to dryness under reduced pressure and then purified by silica gel column chromatography (ethyl acetate / hexane 20% → 55%) 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-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-8) (4.07 g, 81% yield).

[0430] (Condition 3: Lithium diisobutyl-tert-butoxyaluminum hydride: LDBBA) A tetrahydrofuran / hexane solution of LDBBA (0.25 M, 134.2 μL, 33.6 μmol) was added to a tetrahydrofuran suspension (0.45 mL) of compound III-7 (30.0 mg, 30.5 μmol) and molecular sieves 4A (3 mg, 0.1 wt) at -50 °C. After stirring for 1 hour, the reaction mixture was analyzed by HPLC, confirming the progress of the reaction (compound III-7: 5.7% PA, compound III-8: 83.49% PA @ 220 nm, dr = 93.6:6.4).

[0431] (Condition 4: A compound represented by formula A in which three R3s are di-tert-butylmethylphenoxide, hereinafter referred to as "Reagent A") Dibutylhydroxytoluene (885.41 mg, 4.02 mmol) was added to a suspension (2 mL) of lithium aluminum hydride (50.0 mg, 1.32 mmol) in tetrahydrofuran at 0° C., and the mixture was stirred at 25° C. to obtain the compound of the following formula: [ka] Reagent A, designated as the formula (I), was prepared. The prepared solution (80 μL) containing this reagent A was added to a tetrahydrofuran suspension (0.45 mL) of 4-methoxyphenyl 3,6-di-O-benzyl-4-O-{4,6-O-benzylidene-3-O-[(naphthalen-2-yl)methyl]hexopyranosyl-2-ulose}-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-7) (30.0 mg, 30.5 μmol) and molecular sieves 4A (3.0 mg) at 0 °C and stirred for 1 h. The reaction mixture was analyzed by HPLC, confirming the progress of the reaction (Compound III-7: not detected, Compound III-8: 92.3% PA @ 220 nm (excluding BHT), dr = 97.8:2.2).

[0432] (Condition 5: diisobutylaluminum hydride: DIBALH) To a tetrahydrofuran suspension (0.75 mL) of compound III-7 (50.0 mg, 50.8 μmol) and molecular sieves 4A (5 mg, 0.1 wt), a toluene solution of DIBALH (1.5 M, 37 μL, 55.5 μmol) was added at −80° C. After overnight stirring, the mixture was stirred at −60° C. for 90 minutes and at −40° C. for 30 minutes. The progress of the reaction was confirmed by HPLC (compound III-7: ND, compound III-8: 40.92% PA @ 220 nm (excluding toluene), dr = 41.9:58.1).

[0433] 1 H-NMR(500MHz,CDCl3)δ7.79-7.84(m,4H),7.67-7.74(m,4H),7.45-7.50(m, 5H),7.37-7.41(m3H),7.24-7.28(m,4H),7.17-7.21(m,1H),7.03-7.04(m,2H ),6.85-6.93(m,3H),6.81(ddd,J=9.0,4.0,2.0Hz,2H),6.69(ddd,J=9.0,4.0 ,2.0Hz,2H),5.61(d,J=8.5Hz,1H),5.53(s,1H),4.92(d,J=13.0Hz,1H),4.87 (d,J=6.5Hz,1H),4.84(d,J=6.0Hz,1H),4.65-4.68(m,2H),4.48(d,J=12.0H z,1H),4.39-4.46(m,3H),4.14-4.20(m,2H),4.03-4.07(m,2H),3.79(dd,J=1 1.0,3.5Hz,1H),3.70-3.76(m,2H),3.70(s,3H),3.61(t,J=10.0Hz,1H),3.51 (dd,J=10.0,3.5Hz,1H),3.16(td,J=10.0,5.0Hz,1H),2.61(d,J=1.5Hz,1H). 13C-NMR (125MHz,CDCl3)δ155.6,151.0,138.6,137.9,137.7,135.6,134.0,133.5,133.3, 131.8,129.2,128.7,128.4,128.2,128.12,128.09,128.04,127.92,127.89,127. 4, 126.8, 126.4, 126.3, 126.2, 125.9, 123.6, 119.0, 114.6, 101.8, 101.0, 98.0, 78.9, 78.4, 77.7, 77.0, 74.94, 74.91, 73.8, 72.6, 69.9, 68.7, 68.6, 67.0, 55.9, 55.8. HRMS(ESI - )[M+HCO2] - calcd for C 60 H 56 NO 15 :1030.3655;found 1030.3695.

[0434] <Synthesis of Compound III-10> Compound III-10 was synthesized according to the following synthetic scheme 1C. [Synthetic Scheme 1C]

[0435] [ka]

[0436] Example 20: 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-8)

[0437] [ka]

[0438] (Small process 1-12) 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-carboxybenzamido)-2-deoxy-β-D-glucopyranoside (Compound III-6) (6.00 g, 5.98 mmol) in dichloromethane (30 mL), 1-hydroxybenzotriazole monohydrate (0.18 g, 1.20 mmol), N,N-diisopropylethylamine (0.85 g, 6.57 mmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.26 g, 6.57 mmol) were added. The mixture was heated to 40 °C and stirred for 31 hours. After confirming the completion of the reaction by HPLC, the reaction solution was cooled to 0 °C, ethyl acetate (90 mL) and water (60 mL) were added, and 6 N hydrochloric acid was added under vigorous stirring until the pH reached 7. The organic and aqueous layers were separated, and the organic layer was washed with water (60 mL) and saturated brine (30 mL). The solution was concentrated under reduced pressure to a volume of 12 mL. Tetrahydrofuran (60 mL) was added, and the solution was again concentrated to a volume of 9 mL to give a tetrahydrofuran solution of crude 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-8). This product was used directly in the next step.

[0439] Example 21: 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-9)

[0440] [ka]

[0441] (Small process 1-15) A solution of crude 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-8) was added to N,N-dimethylacetamide (60 mL), benzyl bromide (1.53 g, 8.96 mmol), methyl trifluoroacetate (0.15 g, 1.53 g, 1.56 mmol), and methyl ... The mixture was cooled to 0°C, and a solution of lithium tert-butoxide in tetrahydrofuran (a mixture of 2-methyl-2-propanol (0.66 g, 8.96 mmol) and tetrahydrofuran (2.4 mL) was cooled to 0°C, and a solution of n-butyllithium in hexane (1.55 mol / L) (5.78 mL, 8.96 mmol) was added and stirred for 30 minutes) was added and stirred for 3 hours. After confirming the completion of the reaction by HPLC, ethylenediamine (anhydrous) (0.18 g, 2.99 mmol) was added and stirred for an additional 1 hour. Acetic acid (0.72 g, 11.95 mmol) was added to this solution, and the mixture was filtered. The molecular sieves 4A were washed with ethyl acetate (90 mL), and then water (60 mL) was added and the mixture was separated. The organic layer was washed twice with water (60 mL) and then concentrated under reduced pressure until the volume reached 12 mL. Toluene (30 mL) was added, and the mixture was concentrated again until the volume reached 12 mL. Toluene (18 mL) and silica gel 60N (spherical) (9 g) were added, and the mixture was stirred at 25°C 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 9 mL to obtain a toluene solution of crude 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-9).

[0442] Example 22: 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-10)

[0443] [ka]

[0444] (Small process 1-16) The crude 4-methoxyphenyl obtained in Example 21 To a solution of 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-9) (5.50 g (5.48 mmol) based on Compound III-8), dichloromethane (16.5 mL) and molecular sieves 4A (550 mg) were added and cooled to 0 °C. Borane-tetrahydrofuran complex (0.91 mol / L tetrahydrofuran solution) (18.06 mL, 16.43 mmol) and copper(II) trifluoromethanesulfonate (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 the mixture was stirred for an additional 30 minutes. The solution was filtered, and the molecular sieves 4A were washed with ethyl acetate (110 mL). 0.5 N hydrochloric acid (55 mL) was added and the mixture was stirred for 30 minutes. The organic and aqueous layers were separated, and the organic layer was washed with 0.5 N hydrochloric acid (55 mL) and saturated brine (27.5 mL). The solution was concentrated to dryness under reduced pressure. This product was purified by silica gel column chromatography (silica gel 300 g, 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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-10) (4.94 g, yield 83.6%, HPLC area: 98.54%).

[0445] 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). 13 C-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] - calcd for C 67 H 64 NO 15 :1122.4281;found 1122.4285.

[0446] <Synthesis of Compound V-3> Compound V-3 was synthesized according to the following synthetic scheme 2A. [Synthetic Scheme 2A]

[0447] [ka]

[0448] Example 23: 2-O-Acetyl-3,4,6-tri-O-benzyl-1-O-(2,2,2-trichloroethanimidoyl)-D-mannopyranose (Compound IV-3)

[0449] [ka]

[0450] (Small process 2-1&2-2) To a dichloromethane solution (30 mL) of 3,4,6-tri-O-benzyl-1,2-O-(1-methoxyethylidene)-β-D-mannopyranose (Compound IV-1) (3.00 g, 5.92 mmol), p-toluenesulfonic acid monohydrate (5.63 mg, 29.6 μmol) and water (0.3 mL) were added at room temperature and stirred at the same temperature for 90 minutes. After the reaction, triethylamine (0.30 g, 2.96 mmol) was added. After stirring at room temperature for 2 hours, toluene (60 mL) was added. The crude product was concentrated to dryness, and toluene (15 mL) was added to obtain 2-O-acetyl-3,4,6-tri-O-benzyl-D-mannopyranose (Compound IV-2) as a toluene solution. To a mixture of the obtained compound IV-2 in toluene (15 mL) and 2,2,2-trichloroacetonitrile (4.27 g, 29.6 mmol), 1,8-diazabicyclo[5.4.0]-7-undecene (7.1 μL, 47 μmol) was added at room temperature over 30 min. After stirring at the same temperature for 1 h, the reaction mixture was concentrated to dryness. The resulting crude product was purified by silica gel chromatography (Wakogel 9 g, ethyl acetate / hexane 20%) to obtain 2-O-acetyl-3,4,6-tri-O-benzyl-1-O-(2,2,2-trichloroethanimidoyl)-D-mannopyranose (compound IV-3) (3.75 g, 99% yield), which was used in the next step.

[0451] Example 24: 4-Methoxyphenyl 2-O-acetyl-3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[2-O-acetyl-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-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranoside (Compound V-1)

[0452] [ka]

[0453] (Small process 2-3) A solution of 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-4-O-(2,4-di-O-benzyl-β-D-mannopyranosyl) 2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranoside (Compound III-13) (50 mg, 0.02 mmol) and Compound IV-3 (113.4 mg, 0.18 mmol) in dichloromethane (0.8 mL) was added with molecular sieves 4A (10 mg, 0.2 wt) and cooled to -20 °C. After stirring for 15 min, a solution of trimethylsilyl trifluoromethanesulfonate in dichloromethane (0.17 M, 30 μL, 5.0 μmol) was added. After stirring for 90 minutes, triethylamine (5 μL, 0.04 mmol) was added, and the temperature was raised to room temperature (reaction solution 1). To a dichloromethane solution (21 mL) of compound III-13 (650 mg, 0.71 mmol) and compound IV-3 (1.59 g, 2.49 mmol), molecular sieves 4A (140 mg, 0.2 wt) was added and cooled to -20 °C. After stirring for 20 min, a dichloromethane solution of trimethylsilyl trifluoromethanesulfonate (0.28 M, 239 μL, 0.07 mmol) was added, followed by trimethylsilyl trifluoromethanesulfonate (3 μL, 0.02 mmol) 3 h later. After stirring for 45 min, triethylamine (30 μL, 0.21 mmol) was added and the mixture was warmed to room temperature (reaction solution 2). After confirming the progress of the reaction in both reaction solutions 1 and 2, the reaction suspensions were mixed, the molecular sieves were filtered off, and the mixture was washed with dichloromethane (4 mL). The resulting solution was concentrated to dryness, and the resulting crude product (2.71 g) was purified by silica gel chromatography (ethyl acetate / hexane 20% → 55% and ethyl acetate / dichloromethane 0% → 20%) to give compound V-1 (1.16 g, yield 85%).

[0454] 1H-NMR(500MHz,CDCl3)δ2.07(s,3H),2.08(s,3H),3.01(d,J=12Hz,1H),3.27-3.35(m,2H),3.45-3.54(m,2H),3.54-3.94(m,14H),3.73(s,3H),4.00(t,J=9.5Hz,1H),4.09(t,J=8.5Hz,1H),4.26(d,J=12Hz,1H),4.32(d,J=11Hz,1H),4.35-4.64(m,15H),4.68(d,J=7.5Hz,1H),4.71-4.80(m,3H),4.81-4.85(m,2H),4.93(d,J=8.5Hz,1H),4.95(d,J=10Hz,1H),5.14(d,J=8.0Hz,1H),5.17(brs,1H),5.35(brs,1H),5.47(dd,J=2.0, 3.0Hz,1H),6.76(d,J=9.0Hz,2H),6.92(d,J=9.0Hz,2H),7.06-7.38(m,50H). 13 C-NMR (125MHz,CDCl3)δ20.9, 21.0, 66.3, 68.3, 68.57, 68.63, 68.90, 68.92, 71.3, 71.8, 72.2, 73.2, 73.9, 74.1, 74.3, 74.5, 74.7, 74.8, 74.9, 75.3, 77.5, 77.9, 78.0, 78.5, 80.6, 95.6, 97.3, 99.2, 99.6, 99.9, 114.3, 118.5, 127.22, 127.27, 127.31, 127.38, 127.41, 127.45, 127.55, 127.59, 127.64, 127.66, 127.7, 127.8, 128.0, 128.13, 128.16, 128.19, 128.23, 128.29, 128.35, 128.44, 137.6, 137.7, 137.8, 137.96, 137.99, 138.3, 138.40, 138.44, 138.6, 138.9, 151.5, 153.5, 155.1, 170.0, 170.4. MS(ESI)(m / z):1949([M+NH4] + ).

[0455] Example 25: 4-Methoxyphenyl 3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[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-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranoside (Compound V-3)

[0456] [ka]

[0457] (Small process 2-4&2-5) To a solution of compound V-1 (50 mg, 0.03 mmol) in tetrahydrofuran (0.4 mL) were added water (0.05 mL) and a 50% aqueous sodium hydroxide solution (16.6 mg, 0.21 mmol), and the mixture was stirred at 50° C. for 27 hours. Subsequently, methanol (0.05 mL) was added, and the mixture was stirred at 50°C for 18 hours. After cooling to room temperature, acetic acid (8.9 μL, 0.16 mmol) was added to produce a solution containing 4-methoxyphenyl 3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[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-glucopyranoside (Compound V-2). The resulting solution was concentrated to dryness, and then tetrahydrofuran (0.4 mL), water (0.05 mL), sodium bicarbonate (8.7 mg, 0.10 mmol), and 2,2,2-trichloroethyl chloroformate (6.6 mg, 0.03 mmol) were added. After stirring at room temperature for 1 hour, water (2 mL) and ethyl acetate (5 mL) were added to the resulting suspension. After removing the aqueous layer, the crude product was concentrated to dryness and purified by silica gel chromatography (ethyl acetate / hexane 20% → 55% and ethyl acetate / dichloromethane 0% → 20%) to obtain compound V-3 (43.2 mg, 90% yield). Compound V-3 (821.6 mg, 82% yield) was obtained from compound V-1 (1.05 g, 0.54 mmol) in a similar manner.

[0458] MS(ESI)(m / z):1864([M+NH4] + ).

[0459] <Synthesis of Compound V-5> Compound V-5 was synthesized according to the following synthetic scheme 2B. [Synthetic Scheme 2B]

[0460] [ka]

[0461] Example 26: 2-O-Acetyl-3,4,6-tri-O-benzyl-D-mannopyranose (Compound IV-2)

[0462] [ka]

[0463] (Small process 2-1) 3,4,6-Tri-O-benzyl-1,2-O-(1-methoxyethylidene)-β-D-mannopyranose (Compound IV-1) (30.0 g, 59.2 mmol) was added to a 1 L recovery flask and dichloromethane (300 mL) was added. Under nitrogen, toluenesulfonic acid monohydrate (23.0 mg, 0.118 mmol) and water (3.0 mL) were added at room temperature and stirred at the same temperature for 3.5 hours. After confirming the completion of the reaction by HPLC, triethylamine (4.13 mL, 29.6 mmol) was added and stirred at the same temperature for 1.5 hours. After confirming the completion of the Ac group transfer reaction by HPLC, ethyl acetate (450 mL) was added to the reaction mixture and the mixture was separated with 5% aqueous sodium bicarbonate (300 mL). 20% brine (150 mL) was added to the organic layer and the mixture was separated. The organic layer was concentrated under reduced pressure to a volume of 60 mL, and toluene (450 mL) was added and concentrated under reduced pressure to a volume of 60 mL. Anhydrous dichloromethane (150 mL) was added to obtain a colorless solution of 2-O-acetyl-3,4,6-tri-O-benzyl-D-mannopyranose (Compound IV-2) in toluene and dichloromethane. This solution was used directly in the next step.

[0464] Example 27: 2-O-Acetyl-3,4,6-tri-O-benzyl-1-O-(2,2,2-trichloroethanimidoyl)-D-mannopyranose (Compound IV-3)

[0465] [ka]

[0466] (Small process 2-2) A solution of 2-O-acetyl-3,4,6-tri-O-benzyl-D-mannopyranose (compound IV-2) in toluene and dichloromethane (59.2 mmol) was added to a 1 L recovery flask, followed by the addition of trichloroacetonitrile (9.03 mL, 89.0 mmol) and 1,8-diazabicyclo[5.4.0]undec-7-ene (89.0 μL, 0.592 mmol). The mixture was stirred under nitrogen at 0°C for 3 hours. 1,8-diazabicyclo[5.4.0]undec-7-ene (44.5 μL, 0.296 mmol) was added and the mixture was stirred at 0°C for 2.5 hours. 1,8-diazabicyclo[5.4.0]undec-7-ene (44.5 μL, 0.296 mmol) was then added and the mixture was stirred at 0°C for 1 hour. The reaction mixture was then refrigerated overnight. After confirming the completion of the reaction by HPLC, acetic acid (68 μL, 1.18 mmol) was added to the reaction mixture at 0° C. to obtain a brown solution of 2-O-acetyl-3,4,6-tri-O-benzyl-1-O-(2,2,2-trichloroethanimidoyl)-D-mannopyranose (Compound IV-3) in dichloromethane (59.2 mmol). This solution was used directly in the next step.

[0467] Example 28: 4-Methoxyphenyl 2-O-acetyl-3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[2-O-acetyl-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-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound V-4)

[0468] [ka]

[0469] (Small process 2-6) A solution of 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-4-O-(2,4-di-O-benzyl-β-D-mannopyranosyl)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound III-11) (8.70 g, 9.27 mmol) and 2-O-acetyl-3,4,6-tri-O-benzyl-1-O-(2,2,2-trichloroethanimidoyl)-D-mannopyranose (Compound IV-3) in dichloromethane (32.5 mmol) was added to a 300 mL three-neck flask, followed by dichloromethane (15 mL, totaling 131 mL of reaction solution) and molecular sieves 4A powder (1.31 g). Under nitrogen, trimethylsilyl trifluoromethanesulfonate (0.335 mL, 1.86 mmol) was added dropwise over 10 minutes at -20°C, and the mixture was stirred at the same temperature for 3 hours. After confirming the completion of the reaction by HPLC, triethylamine (520 μL, 3.73 mmol) was added and the mixture was stirred at room temperature for 30 minutes. The reaction mixture was filtered through Celite and washed with dichloromethane (43.5 mL). The filtrate was concentrated, and the concentrated residue was purified by column chromatography (silica amount: 650 g, developing solvent: toluene / ethyl acetate = 1 / 0 to 4 / 1). The main fraction was concentrated under reduced pressure to give 4-methoxyphenyl 2-O-acetyl-3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[2-O-acetyl-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-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound V-4) (16.2 g, isolated yield 93.0%) as a white amorphous solid.

[0470] 1H-NMR (500 MHz, CDCl3) δ 7.63 - 6.60 (m, 58H), 5.54 (d, J = 8.4 Hz, 1H), 5.49 (dd, J = 3.2, 2.0 Hz, 1H), 5.34 (brt, J = 3.2 Hz, 1H), 5.13 (brd, J = 1.6 Hz, 1H), 5.05 (d, J = 12.0 Hz, 1H), 4.91 - 4.78 (m, 6H), 4.69 (d, J = 11.2 Hz, 1H), 4.63 - 4.55 (m, 6H), 4.48 - 4.32 (m, 9H), 4.21 (t, J = 11.2 Hz, 2H), 4.07 (t, J = 9.6 Hz, 1H), 4.00 - 3.73 (m, 9H), 3.70 (s, 3H), 3.68 - 3.51 (m, 8H), 3.44 (brd, J = 8.0 Hz, 1H), 3.21 (brd, J = 9.6 Hz, 1H), 2.10 (s, 3H), 1.86 (s, 3H). 13 C-NMR (125 MHz, CDCl3) δ 170.2, 170.0, 155.4, 151.0, 138.9, 138.7, 138.6, 138.5, 138.2, 138.1, 138.0, 137.9, 133.7, 131.7, 128.7, 128.5, 128.4, 128.3, 128.2, 127.9, 127.9, 127.8, 127.7, 127.6, 127.5, 127.5, 127.1, 123.4, 118.8, 114.4, 101.9, 99.8, 98.5, 97.7, 81.3, 79.5, 78.2, 78.0, 77.9, 76.9, 76.6, 75.2, 75.2, 75.1, 74.9, 74.7, 74.5, 74.4, 74.2, 74.2, 73.6, 73.5, 73.4, 72.4, 72.0, 71.9, 71.4, 69.4, 69.4, 68.8, 68.8, 68.3, 66.7, 55.7, 21.2, 20.9. HRMS(ESI + ) [M + NH4] + calcd for C 113 H 119 N2O 25 : 1903.8096; found 1903.8075.

[0471] Example 29: 4-Methoxyphenyl 3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[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-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound V-5)

[0472] [ka]

[0473] (Small process 2-7) 4-Methoxyphenyl 2-O-acetyl-3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[2-O-acetyl-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-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound V-4) (1.0 g, 0.53 mmol) was placed in a 30 mL 2-neck flask, and tetrahydrofuran (5 mL), methanol (3 mL), and methyl trifluoroacetate (53 μL, 0.53 mmol) were added. After stirring at room temperature under nitrogen for 15 minutes, a 1 M solution of potassium tert-butoxide in tetrahydrofuran (0.265 mL, 0.265 mmol) was added dropwise and the mixture was stirred at 50°C for 1 hour. After confirming the completion of the reaction by HPLC, the mixture was cooled to room temperature. Acetic acid (30.3 μL, 0.53 mmol) was added. Ethyl acetate (10 mL) was added, and the mixture was separated three times with 5% brine (10 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated. The concentrated residue was purified by column chromatography (silica amount: 20 g, eluent: toluene / ethyl acetate = 9 / 1 to 7 / 3), and the main fraction was concentrated under reduced pressure to obtain 4-methoxyphenyl 3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[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-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (compound V-5) (900 mg, isolated yield 94%) as a white amorphous substance.

[0474] 1H-NMR(500MHz,CDCl3)δ7.63(brs,2H),7.42(d,J=7.2Hz,2H),7.35-6.66(m,54H),5.57(d,J=8.4Hz,1H), 5.14(brd,J=1.2Hz,1H),5.03(d,J=12.0Hz,1H),4.97(brd,J=1.2Hz,1H),4.93(d,J=13.2Hz,1H), 4.86-4.75(m,3H),4.64-4.33(m,17H),4.21(brt,J=8.8Hz,1H),4.09(brt,J=9.6Hz,1H),3.97-3.51(m,22H),3.43(brd,J=9.6Hz,1H),3.22(brd,J=9.6Hz,1H), 2.34(d,J=2.4Hz,1H),2.19(d,J=2.8Hz,1H). 13 C-NMR (125MHz,CDCl3)δ155.2, 150.8, 138.9, 138.6, 138.5, 138.3, 138.3,138.0, 138.0, 137.8, 137.7, 133.6, 131.6, 128.5, 128.4, 128.4,128.3, 128.2, 128.2, 128.2, 128.0, 127.9, 127.8, 127.8, 127.7,127.7, 127.6, 127.5, 127.4, 127.4, 127.3, 127.3, 126.9, 123.2,118.6, 114.2, 101.6, 101.4, 99.7, 97.6, 81.7, 80.0, 79.7, 79.0,78.2, 76.7, 75.0, 75.0, 74.8, 74.7, 74.6, 74.3, 74.2, 74.2, 74.1,74.1, 73.4, 73.3, 73.2, 72.0, 71.9, 71.4, 71.2, 69.4, 68.8, 68.5,68.1, 67.7, 66.4, 55.5.HRMS(ESI + )[M+NH4] + calcd for C 109 H 115 N2O 23 :1819.7885;found 1819.7886.

[0475] <Synthesis of Compound VI-3> Compound VI-3 was synthesized according to the following synthetic scheme 3A. [Synthetic Scheme 3A]

[0476] [ka]

[0477] Example 30: 4-Methoxyphenyl 2-amino-3,6-di-O-benzyl-2-deoxy-β-D-glucopyranoside (Compound II-2)

[0478] [ka]

[0479] (Small process 3-1) To a solution of 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound II-1) (20.0 g, 33.58 mmol) in n-butanol (80 mL), ethylenediamine (10.09 g, 167.87 mmol) was added, heated to 90 °C, and stirred at the same temperature for 4 hours. After confirming the completion of the reaction by HPLC, the mixture was cooled to 30 °C, and cyclopentyl methyl ether (300 mL) was added. The mixture was stirred at the same temperature for 12 hours. Insoluble matter derived from the phthaloyl group was filtered and washed with cyclopentyl methyl ether (60 mL). The resulting filtrate was washed three times with 25% aqueous methanol (300 mL), and the organic layer was concentrated under reduced pressure to 40 mL. Isopropanol (400 mL) was added to the concentrated solution, which was then concentrated under reduced pressure to 160 mL. Isopropanol (20 mL) and water (130 mL) were added to the concentrated solution, and the temperature was adjusted to 20 °C. Seed crystals of Compound II-2 were added, and the mixture was stirred at the same temperature for 1 hour. After confirming the precipitation of crystals, water (230 mL) was added dropwise over 1 hour. The resulting slurry was cooled to 0-5 °C and stirred at the same temperature for 1 hour. The precipitated crystals were filtered. The filtered crystals were washed with a 0-5 °C mixture of isopropanol and water (40 / 80 mL) and dried under reduced pressure at 40 °C to obtain 4-methoxyphenyl 2-amino-3,6-di-O-benzyl-2-deoxy-β-D-glucopyranoside (Compound II-2) (14.4 g, 92% yield) as white crystals.

[0480] 1 H-NMR(500MHz,CDCl3)δ7.40-7.26(m,10H), 7.01-6.97(m,2H), 6.81-6.76(m,2H), 4.98(d,J=11.5Hz,1H), 4.79(d,J=12.0Hz,1H), 4.69(d,J=8.0Hz,1H), 4.56(q,J=11.5Hz,2H), 3.80-3.74(m,3H), 3.74(s,3H), 3.58-3.52(m,1H), 3.40(t,J=9.5Hz,1H), 3.10(brs,1H), 3.08(dd,J=9.5,8.0Hz,1H), 1.63(brs,2H). 13C-NMR (125MHz,CDCl3)δ155.3, 151.2, 138.5, 137.6, 128.6, 128.4, 127.9, 127.9, 127.8, 127.7, 118.4, 114.4, 103.1, 84.6, 74.9, 73.9, 73.7, 73.1, 70.8, 56.1, 55.6.HRMS(ESI + ) [M+H] + calcd for C 27 H 32 NO6:466.2224;found 466.2239.

[0481] Example 31: 4-Methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranoside (Compound II-3)

[0482] [ka]

[0483] (Small process 3-2) To a solution of 4-methoxyphenyl 2-amino-3,6-di-O-benzyl-2-deoxy-β-D-glucopyranoside (Compound II-2) (10.0 g, 21.48 mmol) in tetrahydrofuran (100 mL), sodium bicarbonate (2.71 g, 32.26 mmol) in water (50 mL) was added, and 2,2,2-trichloroethyl chloroformate (5.01 g, 23.65 mmol) was added dropwise at 25 °C or below, followed by stirring at the same temperature for 30 minutes. After confirming the completion of the reaction by HPLC, ethyl acetate (100 mL) and water (100 mL) were added, and the mixture was separated to obtain the organic layer. The resulting organic layer was washed again with water (100 mL) and then with 20% brine (50 mL). The resulting organic layer was concentrated under reduced pressure to 30 mL, ethyl acetate (150 mL) was added, and the mixture was concentrated under reduced pressure to 50 mL. Heptane (15 mL) was added to the concentrated solution, and seed crystals of Compound II-3 were added. The mixture was stirred at 20°C for 12 hours. After confirming the precipitation of crystals, heptane (150 mL) was added dropwise over 1 hour. The resulting slurry was cooled to 0-5°C and stirred at the same temperature for 1 hour. The precipitated crystals were filtered. The filtered crystals were washed with a mixture of ethyl acetate and heptane (10 / 40 mL) at 0-5°C and dried under reduced pressure at 40°C to obtain 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranoside (Compound II-3) (12.9 g, 94% yield) as white crystals.

[0484] 1 H-NMR(500MHz,CDCl3)δ7.35-7.26(m,10H),6.95-6.91(m,2H),6.76-6.72(m,2H),5.35(d,J=8.0Hz,1H),5.07(d,J=6.9Hz,1H),4.78(dd,J=15 .5,11.5Hz,2H),4.69(brs,2H),4.55(q,J=11.5Hz,2H),3.83(brs,1H),3.79-3.71(m,3H),3.72(s,3H),3.58(brs,2H),2.88(d,J=2.8Hz,1H). 13C-NMR (125MHz,CDCl3)δ155.3, 154.1, 138.1, 137.6, 128.5, 128.4, 128.1, 127.9, 127.8, 127.7, 118.5, 114.4, 99.7, 95.4, 80.2, 74.3, 74.3, 73.9, 73.6, 72.7, 70.3, 57.2, 55.5.HRMS(ESI - )[MH] - calcd for C 30 H 31 Cl3NO8:638.1121;found 638.1099.

[0485] Example 32: 4-Methoxyphenyl 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranoside (Compound II-4)

[0486] [ka]

[0487] (Small process 3-3) Dimethylaminopyridine (0.30 g, 2.46 mmol) and acetic anhydride (2.41 g, 23.61 mmol) were added to a solution of 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranoside (Compound II-3) (10.0 g, 15.60 mmol) in pyridine (40 mL), and the mixture was stirred at 20 °C for 4 hours. After confirming the completion of the reaction by HPLC, the mixture was heated to 50 °C, and ethanol (40 mL) was added dropwise. The mixture was stirred at the same temperature for 30 minutes. After confirming the precipitation of crystals, ethanol (60 mL) was added dropwise at 50 °C over 30 minutes and the mixture was stirred at the same temperature for 1 hour. The slurry was cooled to 25 °C, and water (100 mL) was added dropwise over 1 hour. The slurry was cooled to 0-5°C and stirred at the same temperature for 1 hour. The precipitated crystals were filtered, washed with a 0-5°C mixture of ethanol and water (25 / 25 mL), and dried under reduced pressure at 40°C to obtain 4-methoxyphenyl 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranoside (Compound II-4) (10.4 g, yield 98%) as white crystals.

[0488] 1 H-NMR(500MHz,CDCl3)δ7.35-7.24(m,10H),7.00-6.95(m,2H),6.77-6.73(m,2H),5.44(brs,1H),5.27(d,J=6.9Hz,1H),5.09 (t,J=9.2Hz,1H),4.74-4.62(m,4H),4.50(s,2H),4.19(brs,1H),3.74(s,3H),3.74(brs,1H),3.63-3.56(m,3H),1.91(s,3H). 13 C-NMR (125MHz,CDCl3)δ169.7, 155.4, 153.9, 151.1, 137.8, 137.7, 128.5, 128.3, 127.9, 127.7, 127.6, 118.4, 114.4, 99.1, 77.7, 74.3, 74.1, 73.6, 73.5, 71.4, 69.5, 57.7, 55.6, 20.8.HRMS(ESI - )[MH]- calcd for C 32 H 33 Cl3NO9:680.1226;found 680.1227.

[0489] Example 33: 4-O-Acetyl-3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-glucopyranoside (Compound II-5)

[0490] [ka]

[0491] (Small process 3-4) To a solution of 4-methoxyphenyl 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranoside (Compound II-4) (10.0 g, 14.64 mmol) in dichloromethane (80 mL), hexafluoro-2-propanol (50 mL), and water (5 mL) was added [bis(trifluoroacetoxy)iodo]benzene (8.82 g, 20.51 mmol) at 25 °C or below and stirred at the same temperature for 4 hours. After confirming the completion of the reaction by HPLC, ethyl acetate (250 mL) was added and the mixture was cooled on ice. After that, sodium bicarbonate (5 g) and sodium sulfite (5 g) dissolved in water (100 mL) were added, and the organic layer was separated. The resulting organic layer was washed again with water (100 mL) containing sodium bicarbonate (5 g) and sodium sulfite (5 g), and then with 20% brine (50 mL). The resulting organic layer was concentrated under reduced pressure to 100 mL (crystal precipitation was confirmed during concentration), and heptane (150 mL) was added dropwise. The resulting slurry was cooled to 0-5 °C and stirred at the same temperature for 1 hour. The precipitated crystals were filtered. The filtered crystals were washed with a mixture of ethyl acetate and heptane (8 / 24 mL) at 0-5 °C and dried under reduced pressure at 40 °C to obtain 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-glucopyranoside (Compound II-5) (7.6 g, 90% yield) as white crystals.

[0492] 1 H-NMR(500MHz,CDCl3)δ7.38-7.21(m,10H),5.17(t,J=3.5Hz,1H),5.13(d,J=10.0Hz,1H ),4.98(t,J=10.0Hz,1H).4.78(d,J=12.0Hz,1H),4.65-4.55(m,3H),4.50(dd,J=17.2,12 .0Hz,2H),4.41(d,J=2.9Hz,1H),4.13-4.09(m,1H),3.97(td,J=10.2,2.5Hz,1H),3.72(t ,J=10.0Hz,1H),3.51(dd,J=10.6,7.2Hz,1H),3.44(dd,J=10.0,2.5Hz,1H),1.90(3H,s).13 C-NMR (125MHz,CDCl3)δ169.6, 154.1, 137.8, 137.2, 128.4, 128.1, 127.9, 127.7, 127.7, 95.3, 91.7, 77.3, 74.6, 73.7, 73.5, 70.9, 69.4, 68.8, 54.6, 20.7.HRMS(ESI - )[MH] - calcd for C 25 H 27 Cl3NO8:574.0808;found 574.0834.

[0493] Example 34: 4-O-Acetyl-3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-1-O-[2,2,2-trifluoro-N-phenylethanimidoyl]-β-D-glucopyranoside (Compound II-6)

[0494] [ka]

[0495] (Small process 3-5) A solution of 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-glucopyranoside (Compound II-5) (3.0 g, 5.20 mmol) and molecular sieves 4A powder (powder particle size 10 μm or less) (1.5 g) in dichloromethane (75 mL) was cooled to 0 ° C, and 2,2,2-trifluoro-N-phenylacetimidoyl chloride (1.40 g, 6.74 mmol) and N-methylimidazole (0.68 g, 8.28 mmol) were added dropwise. The mixture was stirred at the same temperature for 15 hours, and the completion of the reaction was confirmed by HPLC. The reaction solution was filtered and washed with dichloromethane (15 mL) to obtain the filtrate. The filtrate cooled to 0-5°C was washed twice with 5% brine (45 mL) at 0-5°C, followed by further washing with 20% brine (15 mL). The resulting organic layer was concentrated under reduced pressure to 9 mL, toluene (60 mL) was added, and the mixture was concentrated under reduced pressure to 30 mL. Toluene (30 mL) was added to the concentrate, and the mixture was again concentrated under reduced pressure to 30 mL. The concentrate was cooled to 0-5°C, and a slurry of neutral silica gel (silica gel 60N, 40-50 μm, 3 g) in toluene (15 mL) cooled to 0-5°C was added. The mixture was stirred at the same temperature for 30 minutes and then filtered. The filtered silica gel was washed with a 1% ethyl acetate-toluene solution (75 mL). The resulting filtrate was concentrated under reduced pressure to 7.5 mL, isopropyl ether (45 mL) was added, and the mixture was seeded with compound II-6 and stirred at 20°C for 1 hour. After confirming the precipitation of crystals, heptane (45 mL) was added dropwise, and the mixture was stirred for 15 hours. The slurry was cooled to 0-5°C and stirred at the same temperature for 1 hour. The crystals were then filtered. The filtered crystals were washed with a 0-5°C mixture of isopropyl ether and heptane (3 / 6 mL) and dried under reduced pressure at 40°C to give 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-1-O-[2,2,2-trifluoro-N-phenylethanimidoyl]-β-D-glucopyranoside (Compound II-6) (3.2 g, 76% yield) as white crystals.

[0496] 1H-NMR(500MHz,CDCl3)δ7.40-7.21(m,12H),7.09(t,J=7.4Hz,1H),6.74(d,J=7.4Hz,2H),6.37(brs,1H),5.26(t,J=9.7Hz,1H),4.80-4.75( m,2H),4.71-4.59(m,3H),4.53(dd,J=15.2,11.7Hz,2H),4.17(brs,1H),3.98(brs,1H),3.85(t,J=10.0Hz,1H),3.57(brs,2H),1.96(s,3H). 13 C-NMR (125MHz,CDCl3)δ169.3, 154.0, 143.0, 137.6, 137.2, 128.7, 128.6, 128.3, 128.2, 128.0, 127.9, 127.9, 127.7, 124.5, 119.1, 95.2, 94.3, 76.1, 74.7, 73.6, 73.5, 71.8, 70.2, 68.6, 53.6, 20.8(one carbon signal is missing).HRMS(ESI - )[MH] - calcd for C 33 H 31 Cl3F3N2O8:745.1104;found 745.1096.

[0497] Example 35: 4-Methoxyphenyl 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranosyl-(1→2)-3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranosyl [bornyl]amino}-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-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranoside (Compound VI-1)

[0498] [ka]

[0499] (Small process 3-6) Molecular sieves 4A (1 mg, 0.1 wt) were added to a dichloromethane solution (0.4 mL) of compound V-3 (10 mg, 5.41 μmol) and compound II-6 (16.2 mg, 0.02 mmol) and then cooled to -40 °C. After stirring for 15 min, a dichloromethane solution of t-butyldimethylsilyl trifluoromethanesulfonate (0.16 M, 10 μL, 1.62 μmol) was added. After stirring for 2 h, triethylamine (10 μL, 0.08 mmol) was added, and the molecular sieves were filtered off at room temperature and washed with dichloromethane (2 mL) (reaction solution 1). Molecular sieves 4A (65 mg, 0.1 wt) were added to a dichloromethane solution (26 mL) of compound V-3 (650 mg, 0.35 mmol) and compound II-6 (1.05 g, 1.41 mmol), followed by cooling to -40 °C. After stirring for 30 min, a dichloromethane solution of t-butyldimethylsilyl trifluoromethanesulfonate (0.4 M, 100 μL, 0.04 mmol) was added. After stirring for 3.5 h, triethylamine (14.7 μL, 0.11 mmol) was added, and the molecular sieves were filtered off at room temperature and washed with dichloromethane (2.6 mL) (reaction solution 2). After confirming the progress of the reaction in both reaction solutions 1 and 2, the two solutions were mixed and concentrated to dryness. The crude product was purified by silica gel chromatography (ethyl acetate / hexane 20% → 40%). The fraction containing the target product was concentrated to dryness. tert-butyl methyl ether (8.5 mL) was added to the mixture, and the precipitated insoluble matter was filtered off. The filter cake was then washed with tert-butyl methyl ether. The resulting solution was concentrated to dryness and purified by preparative HPLC (acetonitrile / water 90% → 100%) to obtain compound VI-1 (860 mg, yield 81%).

[0500] 1H-NMR(500MHz,CD3CN)δ1.95(s,3H),2.00(s,3H),2.84(m,1H),3.05(d,J=9.0Hz,1H),3.20-4.03(m,29H),3.73(s,3H),4.06(brs,1H),4.09(brs,1H),4.15(brs,1H),4.20-5.04(m,40H),5.08(t,J=9.5Hz,1H),5.20(brs,1H),5.73(brs,1H),5.89(d,J=7.0Hz,1H),6.19(d,J=8.0Hz,1H),6.84(d,J=9.0Hz,2H),6.95(d,J=9.0Hz,2H),7.14(t,J=7.5Hz,1H),7.16-7.40(m,67H),7.43(d,J=7.0Hz,2H). 13 C-NMR (125MHz,CD3CN)δ20.1, 21.0, 55.1, 68.5, 68.9, 69.4, 70.1, 70.4, 71.3, 72.1, 72.2, 72.3, 72.4, 72.6, 72.7, 73.1, 73.45, 73.52, 73.68, 73.75, 73.8, 74.1, 74.17, 74.24, 74.28, 75.0, 76.9, 77.7, 78.4, 78.6, 80.5, 95.7, 95.8, 98.0, 99.2, 99.6, 99.8, 100.3, 100.8, 114.3, 117.1, 117.9, 126.3, 127.0, 127.10, 127.15, 127.18, 127.20, 127.27, 127.28, 127.32, 127.37, 127.42, 127.48, 127.51, 127.55, 127.67, 127.72, 127.83, 127.90, 127.95, 128.0, 128.1, 128.2, 128.3, 128.4, 138.01, 138.04, 138.05, 138.07, 138.09, 138.12, 138.36, 138.46, 138.53, 138.7, 138.9, 151.1, 154.1, 154.2, 155.1, 169.2, 169.3. MS(ESI)(m / z):1501([M+2NH4]2+ ).

[0501] Example 36: 4-Methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranosyl-(1→2)-3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}- 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-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranoside (Compound VI-3)

[0502] [ka]

[0503] (Small process 3-7&3-8) To a solution of compound VI-1 (860 mg, 0.29 mmol) in tetrahydrofuran (8.6 mL), water (0.9 mL) and 50% aqueous sodium hydroxide solution (186 mg, 2.3 mmol) were added, followed by stirring for 26 hours at 50° C. Subsequently, 50% aqueous sodium hydroxide solution (46 mg, 0.6 mmol) was added, followed by stirring for 18 hours, followed by addition of methanol (0.86 mL). After stirring for 4 hours, the mixture was cooled to 0°C and then 4N hydrochloric acid was added to adjust the pH of the reaction mixture to 10, yielding a solution containing 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-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-glucopyranoside (Compound VI-2). The resulting solution was concentrated to dryness, and then tetrahydrofuran (8.6 mL), water (0.9 mL), sodium bicarbonate (121.8 mg, 1.45 mmol), and 2,2,2-trichloroethyl chloroformate (202.7 mg, 0.96 mmol) were added. After stirring at room temperature for 1 hour, water (4.3 mL) and ethyl acetate (8.6 mL) were added to the resulting suspension. After removing the aqueous layer, the resulting organic layer was washed twice with 20% brine. The crude product obtained by concentration to dryness was purified by silica gel chromatography (ethyl acetate / hexane 15% → 55%) to give compound VI-3 (593.7 mg, 71% yield).

[0504] MS(ESI)(m / z):1458([M+2NH4] 2+ ).

[0505] <Synthesis of Compound VI-3 or Compound VI-6> Compound VI-3 or compound VI-6 was synthesized according to the following synthetic scheme 3B. [Synthetic Scheme 3B]

[0506] [ka]

[0507] Example 37: 4-Methoxyphenyl 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound II-7)

[0508] [ka]

[0509] (Small process 3-9) To a solution of 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound II-1) (50.0 g, 83.94 mmol) in ethyl acetate (200 mL), triethylamine (11.04 g, 109.12 mmol), dimethylaminopyridine (0.31 g, 2.52 mmol), and acetic anhydride (11.10 g, 109.12 mmol) were added and stirred at 20 ° C for 4 hours. After confirming the completion of the reaction by HPLC, ethanol (500 mL) and water (150 mL) were added dropwise. The slurry was stirred for 1 hour, and the precipitated crystals were filtered. The filtered crystals were washed with a mixture of ethanol and water (150 / 50 mL) and dried under reduced pressure at 40°C to give 4-methoxyphenyl 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound II-7) (49.0 g, yield 91%) as white crystals.

[0510] 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). 13 C-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] + calcd for C 37 H 36 NO9:638.2385;found 638.2401.

[0511] Example 38: 4-O-Acetyl-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-D-glucopyranoside (Compound II-8)

[0512] [ka]

[0513] (Small process 3-10) To a solution of 4-methoxyphenyl 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound II-7) (49.0 g, 76.84 mmol) in dichloromethane (392 mL), hexafluoro-2-propanol (245 mL), and water (25 mL), [bis(trifluoroacetoxy)iodo]benzene (46.3 g, 107.58 mmol) was added at 25 °C or below and stirred at the same temperature for 4 hours. After confirming the completion of the reaction by HPLC, ethyl acetate (1225 mL) was added, the mixture was cooled on ice, and then sodium bicarbonate (24.5 g) and sodium sulfite (24.5 g) dissolved in water (490 mL) were added, and the mixture was separated to obtain an organic layer. The resulting organic layer was washed again with water (490 mL) containing sodium bicarbonate (24.5 g) and sodium sulfite (24.5 g), and then with 20% brine (245 g). The resulting organic layer was concentrated under reduced pressure to 490 mL (crystal precipitation was confirmed during concentration), and heptane (735 mL) was added dropwise. The resulting slurry was cooled to 0-5 °C and stirred at the same temperature for 1 hour. The precipitated crystals were filtered. The filtered crystals were washed with a mixture of ethyl acetate and heptane (39 / 118 mL) at 0-5 °C and dried under reduced pressure at 40 °C to obtain 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-D-glucopyranoside (Compound II-8) (37.5 g, 92% yield) as white crystals.

[0514] 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, 8.0 Hz, 1H), 5.13 (dd, J = 8.4, 10.0 Hz, 1H ), 4.59 (d, J = 12.4 Hz, 1H), 4.54 (s, 2H), 4.50 (dd, J = 8.4, 10.4Hz, 1H), 4.33 (d, J = 12.4 Hz, 1H), 4.17 (dd, J = 8.4, 10.4 Hz,1H), 3.79 (ddd, J = 8.4, 5.2, 4.8 Hz, 1H ), 3.61-3.53 (m, 2H),3.41 (d, J = 8.0 Hz, 1H), 1.93 (s, 3H). 13 C-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 - )[MH] - calcd for C 30 H 28 NO8:530.1820;found 530.1841.

[0515] Example 39: 4-O-Acetyl-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-O-[2,2,2-trifluoro-N-phenylethanimidoyl]-β-D-glucopyranoside (Compound II-9)

[0516] [ka]

[0517] (Small process 3-11) A solution of 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-D-glucopyranoside (Compound II-8) (5.0 g, 9.41 mmol) and molecular sieves 4A powder (powder particle size 10 μm or less) (2.5 g) in dichloromethane (125 mL) was cooled to 0 °C, and 2,2,2-trifluoro-N-phenylacetimidoyl chloride (1.95 g, 9.41 mmol) and N-methylimidazole (0.85 g, 10.35 mmol) were added dropwise. The reaction mixture was stirred at the same temperature for 15 hours. After confirming the completion of the reaction by HPLC, the reaction mixture at 0-5°C was filtered through a column of neutral silica gel (silica gel 60N, 40-50 μm, 15 g) packed with dichloromethane and washed with dichloromethane (125 mL). The filtrate was concentrated to dryness to give 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-O-[2,2,2-trifluoro-N-phenylethanimidoyl]-β-D-glucopyranoside (Compound II-9) (6.0 g, 90% yield) as an oil.

[0518] 1 H-NMR(500MHz,CDCl3)δ7.80-7.69(m,4H),7.37-7.26(m,5H),7.17(t,J=7.7Hz,2H),7.04(t,J=7.5Hz,1H),7.00(d,J=6.5Hz,2H),6.95-6.88(m, 3H),6.59(brs,2H),5.26-5.20(m,1H),4.62(d,J=12.0Hz,1H),4.56(s, 2H),4.49(brs,1H),4.32(d,J=12.0Hz,2H),3.61(brs,2H),1.95(s,3H). 13C-NMR (125MHz,CDCl3)δ169.7, 137.9, 137.6, 134.3, 131.5, 130.5, 129.2, 128.8, 128.6, 128.4, 128.2, 128.0, 127.9, 127.8, 124.6, 123.7, 119.4, 119.3, 76.8, 74.5, 74.2, 73.8, 71.9, 69.1, 54.7, 53.7, 21.1.HRMS(ESI - )[MH] - calcd for C 38 H 32 F3N2O8:701.2116;found 701.2095.

[0519] Example 40: 4-Methoxyphenyl 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranosyl-(1→2)-3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)- Indol-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)-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound VI-4)

[0520] [ka]

[0521] (Small process 3-12) 4-Methoxyphenyl 3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[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-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound V-5) (410 mg, 0.227 mmol) and 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-O-[2,2,2-trifluoro-N-phenylethanimidoyl]-β-D-glucopyranoside (Compound II-9) (800 mg, 1.14 mmol) were added to a 30 mL two-neck flask, followed by dichloromethane (8.2 mL) and molecular sieves 4A powder (61.5 mg). tert-Butyldimethylsilyl trifluoromethanesulfonate (10.4 μL, 45 μmol) was added dropwise over 5 minutes at -65°C under nitrogen, and the mixture was stirred at the same temperature for 1 hour. After confirming the completion of the reaction by HPLC, triethylamine (12.6 μL, 90 μmol) was added and the mixture was stirred at room temperature for 1 hour. The reaction mixture was filtered through Celite and washed with dichloromethane (2.1 mL). The filtrate was separated twice with water (4.1 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated.The concentrated residue was purified by column chromatography (silica amount: 20 g, developing solvent: toluene / ethyl acetate = 9 / 1 to 8 / 2), and the main fraction was concentrated under reduced pressure to give 4-methoxyphenyl 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranosyl-(1→2)-3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-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)-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound VI-4) (534 mg, isolated yield 83%) was obtained as a white amorphous substance.

[0522] 1 H-NMR(500MHz,CDCl3)δ7.76-7.50(m,12H), 7.38-7.59(m,74H),5.51(d,J=8.4Hz,1H),5.07(t,J=8.4Hz,2H),4.95-4.09(m,35H),4.01-3.98(m,3H),3.79(s,2H),3.73-3. 66(m,7H),3.55(brd,J=8.4Hz,1H),3.47-3.16(m,14H),2.88-2.80(m,3H),2.62(m,1H),2.51(m,1H),1.92(s,3H),1.83(s,3H). 13C-NMR (125MHz, CDCl3) δ169.4, 169.3, 155.3, 150.9, 138.8, 138.6, 138.5,138.4, 138.2, 138.0, 138.0, 138.0, 137.7, 133.6, 131.8, 131.6,129.0, 128.9, 128.7, 128.6, 128.4, 128.3, 128.3, 128.3, 128.2,128.1, 128.0, 127.9, 127.8, 127.7, 127.7, 127.5, 127.5, , 74.8, 74.5, 74.2, 74.1, 74.0, 73.7, 73.5,73.4, 73.3, 73.2, 73.0, 72.9, 72.5, 72.4, 72.0, 71.0, 70.8, 70.7,69.8, 69.7, 69.6, 69.4, 68.0, 66.6, 55.6, 5.6, 55.1, 55.0, 20.9, 20.9.HRMS(ESI + )[M+NH4] + calcd for C 169 H 169 N4O 37 :2846.1460;found 2846.1404.

[0523] Example 41: 4-Methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranosyl-(1→2)-3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)- [2,4-di-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-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound VI-5)

[0524] [ka]

[0525] (Small process 3-13) 4-Methoxyphenyl 4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranosyl-(1→2)-3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-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)-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound VI-4) (530 mg, 0.187 mmol) was added to a 30 mL two-neck flask, and tetrahydrofuran (2.1 mL), methanol (1.1 mL), and methyl trifluoroacetate (19 μL, 0.187 mmol) were added. After stirring at room temperature under nitrogen for 20 minutes, a 1 M solution of potassium tert-butoxide in tetrahydrofuran (94 μL, 94 μmol) was added dropwise, and the mixture was stirred at 50 °C for 3 hours. After confirming the completion of the reaction by HPLC, the mixture was cooled to room temperature. Acetic acid (11 μL, 0.187 mmol) was added, and then ethyl acetate (10.3 mL) was added, followed by separation twice with 5% brine (10 mL).The organic layer was dried over sodium sulfate, filtered, and concentrated to give 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranosyl-(1→2)-3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-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)-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound VI-5) (460 mg, isolated yield 90%) was obtained as a white amorphous substance. This product was used directly in the next step without further purification.

[0526] 1 H-NMR(500MHz,CDCl3)δ7.78-7.49(m,12H), 7.38-6.56(m,74H), 5.51(d,J=8.8Hz,1H),4.91-4.28(m,27H),4.24-4.12(m,6H),4.03-3.79(m,10H),3.75-3.64(m,9H),3.57-3.31(m,13H),3. 25(dd,J=10.0,6.4Hz,1H),3.17(d,J=10.0Hz,1H),2.90-2.79(m,3H),2.74(d,J=2.8Hz,1H),2.64-2.57(m,2H),2.40(m,1H).

[0527] Example 42: 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-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-glucopyranoside (Compound VI-2)

[0528] [ka]

[0529] (Small process 3-14) 4-Methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranosyl-(1→2)-3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranosyl-(1→2)-3,4,6-tri-O-benzyl-α-D- A toluene solution (920 μL) of [mannopyranosyl-(1→6)]-2,4-di-O-benzyl-β-D-mannopyranosyl-(1→4)-3,6-di-O-benzyl-2-deoxy-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-β-D-glucopyranoside (Compound VI-5) (460 mg, 0.168 mmol) was added to a 30 mL recovery flask, and 1-butanol (1.2 mL) and ethylenediamine (224 μL) were added. The mixture was stirred at 80°C under nitrogen for 22 hours. After completion of the reaction was confirmed by HPLC, the mixture was cooled to room temperature. Cyclopentyl methyl ether (6.9 mL) was added and the mixture was stirred for 17 hours. The precipitate was filtered off, and the filtrate was separated three times with 25% aqueous methanol (6.9 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated to give 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-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-glucopyranoside (Compound VI-2) (360 mg, isolated yield 92%) as a white amorphous solid. This product was used directly in the next step without further purification.

[0530] 1H-NMR(500MHz,CDCl3)δ7.41-7.04 (m, 76H), 6.96 (d, J = 9.0 Hz, 2H), 6.78 (d, J =9.0 Hz, 2H), 5.15 (s, 1H), 5.09 (d, J = 12.0 Hz, 1H), 4.98-4.83(m, 6H), 4.75-4.69 (m, 3H), 4.64-4.34 (m, 20H), 4.26 (d, J =11.6 Hz, 1H), 4.12 (d, J = 8.4 Hz, 1H), 4.06-3.45 (m, 29H), 3.33-3.21 (m, 6H), 3.04-2.98 (m, 3H), 2.90-2.79 (m, 4H).

[0531] Example 43: 4-Methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranosyl-(1→2)-3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}- 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-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranoside (Compound VI-3)

[0532] [ka]

[0533] (Small process 3-15) 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-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 A cyclopentylmethyl solution (600 μL) of β-D-mannopyranosyl-(1→4)-2-amino-3,6-di-O-benzyl-2-deoxy-β-D-glucopyranoside (Compound VI-2) (300 mg, 0.127 mmol) was added to a 30 mL recovery flask, followed by the addition of tetrahydrofuran (3 mL), water (1.5 mL), and sodium bicarbonate (53 mg, 0.635 mmol). 2,2,2-Trichloroethyl chloroformate (58 μL, 0.419 mmol) was added dropwise over 10 minutes, followed by stirring for 3 hours. After confirming the completion of the reaction by HPLC, ethyl acetate (3 mL) was added and the mixture was separated twice with water (3 mL). The organic layer was washed with 20% brine (1.5 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated. The concentrated residue was purified by column chromatography (silica amount: 20 g, developing solvent: toluene / ethyl acetate = 9 / 1 to 8 / 2), and the main fraction was concentrated under reduced pressure to give 4-methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranosyl-(1→2)-3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[3,6-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-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-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranoside (Compound VI-3) (279 mg, isolated yield 76%) was obtained as a white amorphous substance.

[0534] 1H-NMR(500MHz,CDCl3)δ7.39-6.99(m,70H),6.87(d,J=8.8Hz,2H),6.74(d,J=8.8Hz,2H),5.02(s,1H), 4.91-4.48(m,40H),4.23(d,J=12.8Hz,1H),4.08(s,1H),3.93-3.35(m,34H),3.22(m,4H),2.93(m,1H), 2.75-2.69(m,2H). HRMS(ESI + ) [M+Et3N+H] + calcd for C 156 H 174 Cl9N4O 35 :2982.9124;found 2982.9080.

[0535] The 1H-NMR spectrum was confirmed to be identical to that of compound VI-3 synthesized by a different route.

[0536] Example 44: 4-Methoxyphenyl 3,6-di-O-benzyl-2-deoxy-2-acetylamino-β-D-glucopyranosyl-(1→2)-3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[3,6-di-O-benzyl-2-deoxy-2-acetylamino-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-acetylamino-β-D-glucopyranoside (Compound VI-6)

[0537] [ka]

[0538] (Small process 3-16) 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-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 A cyclopentylmethyl solution (720 μL) of nosyl-(1→4)-2-amino-3,6-di-O-benzyl-2-deoxy-β-D-glucopyranoside (Compound VI-2) (360 mg, 0.153 mmol) was added to a 50 mL recovery flask, and tetrahydrofuran (3.6 mL), triethylamine (0.192 mL, 1.37 mmol), and dimethylaminopyridine (3.7 mg, 31 μmol) were added. Acetic anhydride (101 μL, 1.07 mmol) was added, and the mixture was stirred for 2 hours. HPLC identified the compound of formula VI-7: [ka] After confirming the production of the compound represented by the formula: ethyl acetate (5.4 mL) was added and the mixture was separated with water (3.6 mL). 5% aqueous sodium bicarbonate solution (3.6 mL) was added to the organic layer and the mixture was separated. Water (3.6 mL) was added to the organic layer and the mixture was separated. The organic layer was dried over sodium sulfate, filtered, and concentrated. A solution of the concentrated residue (410 mg) in methanol (5.4 mL) was added to a 50 mL recovery flask. A 28% solution of sodium methoxide in methanol (74 μL, 0.306 mmol) was added and stirred for 3 hours. After confirming the completion of the reaction by HPLC, acetic acid (22 μL, 0.383 mmol) was added and the mixture was concentrated. The concentrated residue was purified by column chromatography (silica amount: 20 g, developing solvent: toluene / ethyl acetate = 3 / 1 to 1 / 1), and the main fraction was concentrated under reduced pressure to obtain 4-methoxyphenyl 3,6-Di-O-benzyl-2-deoxy-2-acetylamino-β-D-glucopyranosyl-(1→2)-3,4,6-tri-O-benzyl-α-D-mannopyranosyl-(1→3)-[3,6-di-O-benzyl-2-deoxy-2-acetylamino-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-acetylamino-β-D-glucopyranoside (Compound VI-6) (250 mg, isolated yield 66%) was obtained as a white amorphous substance.

[0539] 1 H-NMR(500MHz,CDCl3)δ7.38-7.04(m,70H),6.96(d,J=8.8Hz,2H),6.67(d,J=8.8Hz,2H),6.14(d,J=8.0Hz,1H), 5.78(d,J=6.4Hz,1H),5.30(d,J=7.2Hz,1H),5.16(d,J=8.0Hz,1H),5.03-4.18(m,36H),4.07-3.45(m,31H),3.26-3.15(m,2H), 3.07(brs,1H),2.99(m,1H),2.84-2.76(m,2H),2.56(d,J=2.4Hz,1H),1.76-1.59(m,9H). 13C-NMR (125MHz,CDCl3)δ171.7, 170.9, 170.1, 154.9, 151.5, 139.0, 138.8,138.8, 138.7, 138.6, 138.3, 138.2, 138.2, 138.1, 138.0, 137.6,137.5, 128.5, 128.4, 128.3, 128.2, 128.2, 128.1, 128.0, 127.7,127.6, 127.6, 127.5, 127.4, 126.3, 118.0, 114.4, 100.9, 99.3, 99.1, 98.0, 97.4, 80.1, 79.5, 79.3, 78.7, 78.1, 77.9, 77.5, 77.2, 76.9, 75.3, 75.1, 74.8, 74.8, 74.6, 74.4, 74.2, 74.1, 73.9, 73.7, 73.6, 73.5, 73.3, 73.2, 73.0, 72.4, 71.6, 71.2, 71.0, 70.9, 70.6, 69.5, 69.3, 68.9, 66.6, 56.6, 55.5, 29.6, 43.5, 23.4, 23.2. HRMS(ESI + ) [M+H] + calcd for C 147 H 162 N3O 32 :2482.1170;found 2482.1124.

[0540] <Synthesis of Compound IX-5> Compound IX-5 was synthesized according to the following synthetic scheme 4. [Synthetic Scheme 4]

[0541] [ka]

[0542] Example 45: Prop-2-en-1-yl 2,3-di-O-benzoyl-4,6-O-benzylidene-α-D-galactopyranoside (Compound VII-2)

[0543] [ka]

[0544] (Small process 4-1) To a solution of prop-2-en-1-yl 4,6-O-benzylidene-α-D-galactopyranoside (Compound VII-1) (30.0 g, 97.30 mmol) in pyridine (150 mL), benzoyl chloride (47.87 g, 340.54 mmol) was added dropwise at 40°C or below, and the mixture was stirred at the same temperature for 2 hours. After confirming the completion of the reaction by HPLC, the mixture was cooled to 20-30°C, and ethanol (450 mL) was added, followed by the dropwise addition of water (300 mL) over 30 minutes. After stirring the slurry at 20-30°C for 1 hour, the precipitated crystals were filtered, washed with a mixture of ethanol and water (75 / 75 mL), and dried under reduced pressure at 40°C to obtain prop-2-en-1-yl 2,3-di-O-benzoyl-4,6-O-benzylidene-α-D-galactopyranoside (Compound VII-2) (47.9 g, yield 95%) as white crystals.

[0545] 1 H-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] + calcd for C 30 H 29 O8:517.1857;found 517.1880.

[0546] Example 46: Prop-2-en-1-yl 2,3-di-O-benzoyl-α-D-galactopyranoside (Compound VII-3)

[0547] [ka]

[0548] (Small process 4-2) A solution of prop-2-en-1-yl 2,3-di-O-benzoyl-4,6-O-benzylidene-α-D-galactopyranoside (Compound VII-2) (47.1 g, 91.18 mmol) in acetonitrile (377 mL) was heated to 45 °C, and water (24 mL) and concentrated hydrochloric acid (9.2 g, 91.18 mmol) were added. The mixture was stirred at the same temperature for 30 minutes. Water (353 mL) was added dropwise over 3 hours at 45-50 °C, followed by stirring for an additional 30 minutes. After confirming the completion of the reaction by HPLC, sodium acetate (11.22 g, 136.78 mmol) was added, and ethyl acetate (942 mL) and water (471 mL) were added. The mixture was cooled to below 25 °C and separated to obtain the organic layer. The resulting organic layer was washed twice with water (471 mL) and then with 20% brine (236 mL). The organic layer was concentrated under reduced pressure to 141 mL, toluene (707 mL) was added, and the mixture was concentrated under reduced pressure again to 141 mL. Toluene (236 mL) was added to the resulting concentrate, and the mixture was concentrated under reduced pressure to 141 mL. The concentrate was cooled to 0-5 °C, and a toluene (330 mL) slurry containing neutral silica gel (silica gel 60N, 40-50 μm, 141 g) cooled to 0-5 °C was added. The mixture was stirred at the same temperature for 15 minutes to adsorb the product onto the silica gel, followed by filtration. The silica gel solid phase containing the product was washed with toluene (942 mL) at 0-5 °C (the filtrate from the toluene wash was discarded), and the target compound was desorbed from the silica gel with cyclopentyl methyl ether (707 mL) to yield a cyclopentyl methyl ether solution of prop-2-en-1-yl 2,3-di-O-benzoyl-α-D-galactopyranoside (Compound VII-3) (quantitative weight 36.2 g, quantitative yield 93%). This solution was used in the next step.

[0549] Example 47: Methyl 5-acetamido-3,5-dideoxy-D-glycero-D-galacto-non-2-ulopyranosonate monohydrate (Compound VIII-2)

[0550] [ka]

[0551] (Small process 4-3) To a solution of 5-acetamido-3,5-dideoxy-D-glycero-D-galacto-non-2-ulopyranosonic acid (Compound VIII-1) (40.1 g, 129.66 mmol) and methyl orthoformate (15.60 mL, 142.59 mmol) in methanol (321 mL), sulfuric acid (1.0 g, 10.20 mmol) was added, heated to 40 °C, and stirred for 3 hours. After confirming the completion of the reaction by HPLC, the mixture was cooled to 25 °C, dimethylacetamide (40 mL) was added, and the mixture was concentrated under reduced pressure to 160 mL. The resulting concentrate was adjusted to 15 °C, and water (20 mL) and ethyl acetate (722 mL) were added. After stirring at 25 °C for 1 hour, the slurry was cooled to 0-5 °C and stirred at the same temperature for 2 hours. The precipitated crystals were filtered, washed with ethyl acetate (80 mL) at 0-5°C, and dried under reduced pressure at 40°C to obtain methyl 5-acetamido-3,5-dideoxy-D-glycero-D-galacto-non-2-ulopyranosonate monohydrate (Compound VIII-2) (41.1 g, yield 93%) as white crystals.

[0552] 1 H-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). 13 C-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] + calcd for C 12 H 22 NO9:324.1289;found 324.1288.

[0553] Example 48: Methyl 5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-D-glycero-D-galacto-non-2-ulopyranosonate (Compound VIII-3)

[0554] [ka]

[0555] (Small process 4-4) A slurry of methyl 5-acetamido-3,5-dideoxy-D-glycero-D-galacto-non-2-ulopyranosonate monohydrate (Compound VIII-2) (40.3 g, 118.07 mmol) in acetonitrile (403 mL) was adjusted to 25 °C, and acetic anhydride (60.27 g, 590.36 mmol) and paratoluenesulfonic acid monohydrate (1.12 g, 5.89 mmol) were added and stirred at 25 °C for 24 hours. The reaction mixture was then cooled to 15 °C, and acetic anhydride (12.05 g, 118.03 mmol) was added. The mixture was stirred at the same temperature for 47 hours. After confirming the completion of the reaction by HPLC, methanol (40 mL) was added, the temperature was adjusted to 25 °C, and the mixture was stirred at the same temperature for 2 hours. Next, sodium acetate (0.97 g, 11.82 mmol) was added, and the mixture was stirred at the same temperature for an additional 1 hour. The reaction mixture was concentrated under reduced pressure to 120 mL and cooled to 0-5°C. Ethyl acetate (403 mL) and water (161 mL) were added, and triethylamine was added with stirring at 0-5°C to adjust the pH to 7.0. The organic layer obtained by separation was washed twice with 10% brine (121 mL) and concentrated under reduced pressure to 200 mL. Ethyl acetate (605 mL) was added to the concentrated mixture, and the mixture was again concentrated under reduced pressure to 200 mL. Ethyl acetate (40 mL) was added to the concentrated mixture, seed crystals were added, and the mixture was stirred at 25°C for 4 hours. Heptane (302 mL) was then added dropwise over 30 minutes. After stirring the slurry at 25°C for 2 hours, the precipitated crystals were filtered, washed with a mixture of ethyl acetate and heptane (67 / 135 mL), and dried under reduced pressure at 35°C to obtain methyl 5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-D-glycero-D-galacto-non-2-ulopyranosonate (Compound VIII-3) (44.1 g, yield 76%) as white crystals.

[0556] 1 H-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). 13 C-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] + calcd for C 20 H 30 NO 13 :492.1712;found 492.1712.

[0557] Example 49: Methyl 5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-2-O-(2,2,2-trifluoro-N-phenylethanimidoyl)-D-glycero-β-D-galacto-non-2-ulopyranosonate (Compound VIII-4)

[0558] [ka]

[0559] (Small process 4-5) A slurry of methyl 5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-D-glycero-D-galacto-non-2-ulopyranosonate (compound VIII-3) (44.0 g, 89.53 mmol) and molecular sieves 4A powder (powder particle size 10 μm or less) (22 g) in dichloromethane (352 mL) was adjusted to 20 ° C. and stirred at the same temperature for 30 minutes. 2,2,2-trifluoro-N-phenylacetimidoyl chloride (26.02 g, 125.35 mmol) was then added. Subsequently, N-methylimidazole (11.03 g, 134.33 mmol) was added dropwise and stirred at 20 ° C. for 7.5 hours. After confirming the completion of the reaction by HPLC, the reaction solution was filtered and washed with dichloromethane (88 mL) to obtain the filtrate. The resulting filtrate was cooled to 0°C, and cold water (440 mL) was added. Triethylamine was added while stirring at 0-5°C to adjust the pH to 7.5. After stirring at 0-5°C for 30 minutes, the layers were separated. The resulting organic layer was washed twice with cold water (440 mL) and with chilled 20% brine (220 mL) and concentrated under reduced pressure to 88 mL. Ethyl acetate (440 mL) was added to the concentrate, and the mixture was again concentrated under reduced pressure to 88 mL. tert-Butyl methyl ether (308 mL) was added to the concentrate, seed crystals were added, and the mixture was stirred at 20°C for 4 hours. Heptane (264 mL) was added dropwise to the resulting slurry over 1 hour, and the mixture was stirred at the same temperature for 2 hours. The precipitated crystals were then filtered, washed with a mixture of t-butyl methyl ether and heptane (132 / 88 mL), and dried under reduced pressure at 35°C to obtain methyl 5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-2-O-(2,2,2-trifluoro-N-phenylethanimidoyl)-D-glycero-β-D-galacto-non-2-ulopyranosonate (Compound VIII-4) (39.5 g, yield 67%) as white crystals.

[0560] 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). 13 C-NMR (125MHz,CDCl3)δ171.0, 170.7, 170.4, 170.2, 170.1, 165.3, 142.6, 141.0(q, 2 J C-F =36.0Hz), 128.8, 124.6, 119.0, 115.1(q, 1 J C-F HRMS(ESI + )[M+NH4] + calcd for C 28 H 37 F3N3O 13 :680.2273;found 680.2314.

[0561] Example 50: Methyl 4,7,8,9-tetra-O-acetyl-5-[acetyl(tert-butoxycarbonyl)amino]-3,5-dideoxy-2-O-(2,2,2-trifluoro-N-phenylethanimidoyl)-D-glycero-β-D-galacto-non-2-ulopyranosonate (Compound VIII-5)

[0562] [ka]

[0563] (Small process 4-6) To a solution of methyl 5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-2-O-(2,2,2-trifluoro-N-phenylethanimidoyl)-D-glycero-β-D-galacto-non-2-ulopyranosonate (Compound VIII-4) (39.0 g, 58.86 mmol) in tetrahydrofuran (390 mL), di-tert-butyl dicarbonate (27.05 g, 123.94 mmol) and dimethylaminopyridine (1.80 g, 14.73 mmol) were added and the mixture was heated to reflux. After stirring at reflux for 30 minutes, completion of the reaction was confirmed by HPLC, and the reaction mixture was concentrated under reduced pressure to 117 mL. Toluene (195 mL) was added to the concentrated mixture, and the mixture was again concentrated under reduced pressure to 117 mL. The concentrated solution was filtered through a silica gel-filled funnel (silica gel 60N, 40-50 μm, 117 g, wet-packed with toluene) and washed with a toluene / ethyl acetate mixture (8 / 2) (975 mL) to obtain a filtrate. The filtrate was concentrated under reduced pressure (to a weight of 59 g), and cyclopentyl methyl ether (23 mL) was added. The solution was adjusted to 20°C, and heptane (156 mL) was added dropwise over 15 minutes. The mixture was stirred at the same temperature for 1 hour. After confirming the precipitation of crystals, heptane (312 mL) was added dropwise over 1 hour, and the precipitated crystals were filtered. The filtered crystals were washed with heptane (78 mL) and dried under reduced pressure at 35°C to obtain methyl 4,7,8,9-tetra-O-acetyl-5-[acetyl(tert-butoxycarbonyl)amino]-3,5-dideoxy-2-O-(2,2,2-trifluoro-N-phenylethanimidoyl)-D-glycero-β-D-galacto-non-2-ulopyranosonate (Compound VIII-5) (37.0 g, 82% yield) as white crystals.

[0564] 1 H-NMR (500 MHz, CDCl3) Note: Detected as a mixture of ca. 1 / 4 isomers. Major isomer: δ 7.27 (t, J = 8.9 Hz, 2H), 7.09 (t, J = 7.2 Hz, 1H), 6.73 (d, J = 8.0 Hz, 2H), 5.75 - 5.65 (m, 1H), 5.31 (d, J = 4.6 Hz, 1H), 5.18 - 5.14 (m, 1H), 5.15 (d, J = 6.0 Hz, 2H), 4.54 (dd, J = 12.0, 2.0 Hz, 1H), 4.08 (dd, J = 12.6, 6.9 Hz, 1H), 3.84 (s, 3H), 2.90 (dd, J = 13.7, 5.2 Hz, 1H), 2.39 (s, 3H), 2.25 (dd, J = 13.7, 11.2 Hz, 1H), 2.09 (s, 3H), 2.07 (s, 3H), 1.99 (s, 3H), 1.77 (s, 3H), 1.62 (s, 9H). Minor isomer: δ inter alia 6.76 (d, J = 8.0 Hz, 2H), 5.85 - 5.80 (m, 1H), 5.29 - 5.25 (m, 1H), 5.22 - 5.19 (m, 1H), 4.44 (d, J = 11.0 Hz, 1H), 4.15 - 4.11 (m, 1H), 3.03 (dd, J = 14.0, 5.0 Hz, 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). 13 C-NMR (125 MHz, CDCl3) Mixture: δ 173.7, 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] + calcd for C 33 H 45 F3N3O 15 : 780.2797; found 780.2801.

[0565] Example 51: Prop-2-en-1-yl 2,3-di-O-benzoyl-6-O-{4,7,8,9-tetra-O-acetyl-5-[acetyl(tert-butoxycarbonyl)amino]-3,5-dideoxy-1-methyl-D-glycero-α-D-galacto-non-2-ulopyranosyl}-α-D-galactopyranoside (Compound IX-1)

[0566] [ka]

[0567] (Small process 4-7) A solution of prop-2-en-1-yl 2,3-di-O-benzoyl-α-D-galactopyranoside (Compound VII-3) in cyclopentyl methyl ether (quantitative value: 31.46 g, 73.43 mmol) was concentrated under reduced pressure to 105 mL, and then added to a solution of methyl 4,7,8,9-tetra-O-acetyl-5-[acetyl(tert-butoxycarbonyl)amino]-3,5-dideoxy-2-O-(2,2,2-trifluoro-N-phenylethanimidoyl)-D-glycero-β-D-galacto-non-2-ulopyranosonate (Compound III-5) (35.0 g, 45.89 mmol) in cyclopentyl methyl ether (175 mL). Next, cyclopentyl methyl ether was added to the resulting mixture, and the total volume was adjusted to 350 mL (a cyclopentyl methyl ether mixed solution of Compound VII-3 and Compound III-5). Cyclopentyl methyl ether (525 mL) and molecular sieves 4A powder (powder particle size 10 μm or less) (17.5 g) were added to a separate vessel and cooled to -60 °C. Then, trimethylsilyl trifluoromethanesulfonate (4.2 mL, 23.24 mmol) was added. To this solution, a cyclopentyl methyl ether mixture of Compound VII-3 and Compound III-5 was added dropwise over 4.5 hours at -60 °C under vigorous stirring, followed by stirring at the same temperature for 2 hours. After confirming the completion of the reaction by HPLC, triethylamine (4.5 mL, 32.12 mmol) was added, and the reaction mixture was warmed to 0 °C. Celite 545 (35.00 g) was then added, and the reaction mixture was filtered and washed with cyclopentyl methyl ether (175 mL). Water (350 mL) was added to the filtrate, and the mixture was separated. Next, 0.5 N aqueous hydrochloric acid (350 mL) was added to the organic layer, and the mixture was stirred at 20 °C for 2 hours. After confirming the decomposition of by-products by HPLC, the organic layer was separated to obtain the organic layer. The organic layer was washed with water (350 mL) and 20% brine (175 mL) and then concentrated under reduced pressure to 70 mL. Toluene (700 mL) was added to the concentrated solution, and the mixture was concentrated under reduced pressure to 70 mL. Toluene (700 mL) and neutral silica gel (silica gel 60N, 40-50 μm, 158 g) were added to the concentrated solution again, and the mixture was stirred at 20°C for 30 minutes.The product was adsorbed onto silica gel, filtered, and the silica gel solid phase containing the product was washed with toluene (1575 mL) (the filtrate from the toluene wash was discarded). The target product was then desorbed from the silica gel with ethyl acetate (875 mL). The resulting ethyl acetate solution was concentrated under reduced pressure to 70 mL, and toluene (175 mL) was added. The mixture was again concentrated under reduced pressure to 70 mL 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(tert-butoxycarbonyl)amino]-3,5-dideoxy-1-methyl-D-glycero-α-D-galacto-non-2-ulopyranosyl}-α-D-galactopyranoside (Compound IX-1). This solution was used in the next step.

[0568] Example 52: 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-ulopyranosyl}-2,3-di-O-benzoyl-α-D-galactopyranoside (Compound IX-2)

[0569] [ka]

[0570] (Small process 4-8) 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(tert-butoxycarbonyl)amino]-3,5-dideoxy-1-methyl-D-glycero-α-D-galacto-non-2-ulopyranosyl}-α-D-galactopyranoside (Compound IX-1) obtained as described above, dichloromethane (525 mL) and copper(II) trifluoromethanesulfonate (8.30 g, 22.95 mmol) were added, the mixture was heated to 40°C, and stirred at the same temperature for 2 hours. After confirming the completion of the reaction by HPLC, the mixture was cooled to 25°C and concentrated under reduced pressure to 70 mL. Ethyl acetate (525 mL) was added to the concentrated solution, which was then washed three times with 5% brine (350 mL). Heptane (263 mL) was added to the organic layer, which was then washed four times with 20% aqueous methanol (525 mL). After confirming by HPLC that impurities derived from the β-eliminated form of compound 8, a by-product of the glycosylation reaction, had been removed from the aqueous layer, the organic layer was concentrated under reduced pressure to 70 mL. Isopropenyl acetate (525 mL) was added to the concentrated solution, which was then concentrated under reduced pressure to 350 mL to obtain a 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-ulopyranosyl}-2,3-di-O-benzoyl-α-D-galactopyranoside (Compound IX-2) in isopropenyl acetate. This solution was used in the next step.

[0571] Example 53: 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-non-2-ulopyranosyl]-α-D-galactopyranoside (Compound IX-3)

[0572] [ka]

[0573] (Small process 4-9) To a 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-ulopyranosyl}-2,3-di-O-benzoyl-α-D-galactopyranoside (Compound IX-2) prepared as described above in isopropenyl acetate, paratoluenesulfonic acid monohydrate (0.88 g, 4.62 mmol) was added, the mixture was heated to reflux (internal temperature around 90 °C), and stirred at the same temperature for 3 hours. After confirming the completion of the reaction by HPLC, the mixture was cooled to 25 °C, triethylamine (0.95 mL, 6.85 mmol) was added, and the mixture was concentrated under reduced pressure to 70 mL. Toluene (350 mL) was added to the concentrated solution, and the mixture was again concentrated under reduced pressure to 70 mL. Toluene (630 mL) was added to the concentrated solution, and neutral silica gel (silica gel 60N, 40-50 μm, 123 g) was added, and the mixture was stirred at the same temperature for 30 minutes to adsorb the product onto the silica gel, followed by filtration. The silica gel solid phase containing the product was washed with toluene (1925 mL) and a mixture of toluene and ethyl acetate (97 / 3, 1400 mL) (the filtrate from the washings was discarded). The target compound was then desorbed from the silica gel solid phase containing the product with ethyl acetate (1050 mL). The resulting ethyl acetate solution was concentrated under reduced pressure to give 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-non-2-ulopyranosyl]-α-D-galactopyranoside (Compound IX-3) (30.1 g, 67% yield (based on Compound VIII-5)) as a white foamy solid (containing 0.42 equivalents of toluene (approximately 4 wt%)).

[0574] 1H-NMR(500MHz,CDCl3)δ7.99 (d, J = 8.4 Hz, 2H), 7.88 (dd, J = 8.4, 1.4 Hz, 2H), 7.53-7.47 (m, 2H), 7.37 (dt, J = 14.7, 6.9 Hz, 4H), 5.90-5.82 (m, 1H), 5.82 (dd, J = 10.9, 3.4 Hz, 1H), 5.73 (d, J = 2.9 Hz,1H), 5.58 (dd, J = 10.9, 4.0 Hz, 1H), 5.51 (td, J = 10.6, 5.0 Hz, 1H), 5.35-5.30 (m, 3H), 5.17-5.15 (m, 2H), 4.94 (dd, J = 10.3, 1.7 Hz,1H), 4.38-4.27 (m, 3H), 4.20-4.13 (m, 2H), 4.08 (dd, J =13.2, 5.7 Hz,1H), 3.94 (dd, J = 10.3, 6.3 Hz,1H), 3.82 (s, 3H), 3.50 (dd, J = 9.7, 7.4 Hz,1H), 2.73 (dd,J = 13.2, 5.2 Hz,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, 10.9 Hz,1H). 13 C-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]+ calcd for C 47 H 56 NO 22 :986.3288;found 986.3277.

[0575] The spectrum of the obtained compound was confirmed to match that of the following literature: Reference 4) J. Org. Chem. 2016, 81, 10600-10616.

[0576] Example 54-1: 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-ulopyranosyl]-D-galactopyranose (Compound IX-4)

[0577] [ka]

[0578] (Small process 4-10) A solution 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-α-D-galacto-non-2-ulopyranosyl]-α-D-galactopyranoside (Compound IX-3) (29.00 g, 29.41 mmol), 1,3-dimethylbarbituric acid (9.19 g, 58.86 mmol), and triphenylphosphine (2.31 g, 8.81 mmol) in methanol (290 mL) was degassed by vacuum and nitrogen substitution five times. Palladium(II) acetate (0.66 g, 2.94 mmol) was then added and the mixture was stirred at 40°C for 12 hours. After confirming the completion of the reaction by HPLC, toluene (580 mL) and water (1015 mL) were added and the mixture was separated to obtain the organic layer. The organic layer was washed four times with 20% aqueous methanol (580 mL). After removing the 1,3-dimethylbarbituric acid, the mixture was concentrated under reduced pressure to 58 mL. Toluene (435 mL) was added and the mixture was again concentrated under reduced pressure to 58 mL. Toluene (383 mL), chloroform (197 mL), and neutral silica gel (silica gel 60N, 40-50 μm, 145 g) were added to the concentrated solution and stirred for 30 minutes to adsorb the product onto the silica gel, followed by filtration. The silica gel solid phase containing the product was washed with a toluene / chloroform mixture (2 / 1, 4350 mL) (the filtrate from the washings was discarded), and the target compound was desorbed from the silica gel solid phase containing the product with ethyl acetate (870 mL). To the resulting ethyl acetate solution, SH silica gel (29.00 g) was added, stirred for 30 minutes, filtered, and washed with ethyl acetate (145 mL) to obtain an ethyl acetate solution containing the target compound. The resulting solution was concentrated under reduced pressure to 58 mL, and toluene (145 mL) was added, followed by another concentration under reduced pressure to 58 mL. The concentrated solution was purified using a silica gel column (silica gel 60N, 40-50 μm, 290 g, mobile phase: hexane / ethyl acetate 50 / 50 to 30 / 70), and selected fractions were concentrated under reduced pressure to 29 mL. Ethyl acetate (290 mL) and activated carbon (Shirasagi A, 14.5 g) were added to the concentrated solution, stirred for 30 minutes, filtered, and washed with ethyl acetate (87 mL) to obtain a purified ethyl acetate solution containing the target compound.The resulting solution was concentrated under reduced pressure 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-ulopyranosyl]-D-galactopyranose (Compound IX-4) (18.70 g, 67% yield) as a white foamy solid.

[0579] 1 H-NMR(500MHz,CDCl3)Major isomer:δ7.98-8.03 (m, 2H), 7.89 (t, 2H, J = 8.9 Hz), 7.52-7.48 (m, 2H), 7.39-7.34 (m, 4H), 5.92 (dd, 1H, J = 10.3, 2.9 Hz), 5.82 (d, 1H, J = 3.4Hz), 5.68 (t, 1H, J = 2.3 Hz), 5.59-5.48 (m, 2H), 5.37 (td, 1H, J = 7.5,2.5 Hz), 5.17 (d, 1H, J =7.5 Hz), 5.02 (d, 1H, J = 10.5 Hz), 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, 6.0 Hz), 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)α / β 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] - calcd for C 45 H 52 NO 24 :990.2885;found 990.2873.

[0580] Example 54-2: Purification method for 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-ulopyranosyl]-D-galactopyranose (Compound IX-4) by crystallization

[0581] [ka]

[0582] 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-ulopyranosyl]-D-galactopyranose (Compound IX-4) (3.00 g, 3.17 mmol, sialyl moiety α / β ratio = 95.7 / 4.3) was dissolved in ethyl acetate (4 mL), followed by addition of 2-propanol (60 mL). The mixture was stirred at 25°C and then concentrated under reduced pressure to 18 mL. The slurry was stirred at 0°C for 3 hours, and the precipitated crystals were filtered. The filtered crystals were washed with cold 2-propanol (9 mL) and dried under reduced pressure at 40 °C 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-ulopyranosyl]-D-galactopyranose (Compound IX-4) (2.66 g, yield 88.7%, sialyl moiety α / β ratio = >99.9 / ND) as white crystals.

[0583] [Analysis conditions] Column: CAPCELL PAK ADME φ4.6 × 150 mm, film thickness 3 μm Wavelength: 220nm Oven: 40°C Eluent: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile Gradient: 0-150min (B) conc. 40% 150.1min (B) conc.95% 155min (B) conc.95% 155.1min(B)conc.40% 160min (B) conc.40% Flow rate: 1mL / min Injection: 5μL Sialyl α-body: 131.0 min, sialyl β-body: 126.2 min

[0584] Example 55: 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-ulopyranosyl]-1-O-(2,2,2-trifluoro-N-phenylethanimidoyl)-D-galactopyranose (Compound IX-5)

[0585] [ka]

[0586] (Small process 4-11) To a solution 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-α-D-galacto-non-2-ulopyranosyl]-D-galactopyranose (Compound IX-4) (42.9 g, 45...

Claims

1. The following formula III-5: 【Chemical 1】 (In the formula, X ２ is an acetyl group), the method comprising the step of producing a compound represented by the following formula III-4: 【Chemistry 2】 (In the formula, X １ represents a substituent selected from the group consisting of a trifluoromethanesulfonyl group, a nonafluorobutanesulfonyl group, a 2-nitrobenzenesulfonyl group, and a 4-nitrobenzenesulfonyl group) with cesium acetate or tetrabutylammonium acetate to produce a compound of formula III-5.

2. Formula IX-1 below: 【Chemistry 3】 A method for producing a compound represented by the following formula VIII-5: 【Chemistry 4】 and a compound represented by the following formula VII-3: 【Chemistry 5】 by α-2,6-glycosidic bonding to form a compound represented by formula IX-1.

3. Formula II-5 below: 【Chemistry 6】 or a compound of formula II-8: 【Chemistry 7】 A method for producing a compound represented by the formula: In fluorous alcohol and water, the following formula II-4: 【Chemistry 8】 or a compound of formula II-7: 【Chemistry 9】 with λ3-iodane to produce a compound represented by formula II-5 or a compound represented by formula II-8.

4. 4. The method of claim 3, wherein the λ3-iodane is selected from the group consisting of [bis(trifluoroacetoxy)iodo]benzene (PIFA), [hydroxy(tosyloxy)iodo]benzene (HTIB), (diacetoxyiodo)benzene (PIDA), [bis(trifluoroacetoxy)iodo]pentafluorobenzene, [hydroxy(methanesulfonyloxy)iodo]benzene, and combinations thereof.

5. 5. The method of claim 3, wherein the fluorous alcohol is selected from the group consisting of hexafluoro-2-propanol (HFIP), 2,2,2-trifluoroethanol (TFE), 2,2,3,3,4,4,5,5-octafluoro-1-pentanol, nonafluoro-tert-butyl alcohol, and combinations thereof.

6. Formula II-6 below: 【Chemistry 10】 or a compound of the following formula II-9: 【Chemistry 11】 The method for producing a compound represented by the following formula II-5: 【Chemistry 12】 or a compound of formula II-8: 【Chemistry 13】 is reacted with a compound of the following formula: 【Chemistry 14】 with 2,2,2-trifluoro-N-phenylacetimidoyl chloride (TFPC) of formula II-6 to produce a compound of formula II-9.

7. The following formula III-6: 【Chemistry 15】 A compound represented by the formula:

8. The following formula III-10: 【Chemistry 16】 A compound represented by the formula:

9. The following formula VIII-5: 【Chemistry 17】 A compound represented by the formula:

10. The following formula VIII-4: 【Chemistry 18】 A method for producing a compound represented by the following formula VIII-3: 【Chemistry 19】 is reacted with a compound of the following formula: 【Chemistry 20】 with 2,2,2-trifluoro-N-phenylacetimidoyl chloride (TFPC) of formula VIII-4 to produce a compound of formula VIII-4.

11. The following formula III-9: 【Chemical 21】 The method for producing a compound represented by the following formula III-8: 【Chemical 22】 by protecting the hydroxyl group attached to the carbon atom at position 2 of the D-mannopyranoside with a benzyl group to produce a compound represented by formula III-9.

12. Formula IX-1 below: 【Chemical 23】 The method for producing the compound represented by formula IX-1 comprises a step of contacting a solvent in which the compound represented by formula IX-1 is dissolved with silica gel to perform solid phase extraction of the compound represented by formula IX-1.

13. 13. The method of claim 12, wherein the solvent in which the compound of formula IX-1 is dissolved is selected from the group consisting of toluene, dichloromethane, chloroform, and combinations thereof.

14. Formula IX-3 below: 【Chemistry 24】 The method for producing the compound represented by formula IX-3 comprises a step of contacting a solvent in which the compound represented by formula IX-3 is dissolved with silica gel to perform solid phase extraction of the compound represented by formula IX-3.

15. 15. The method of claim 14, wherein the solvent in which the compound of formula IX-3 is dissolved is selected from the group consisting of toluene, dichloromethane, chloroform, and combinations thereof.

16. Formula VII-3 below: 【Chemistry 25】 The method for producing the compound represented by formula VII-3 comprises a step of contacting a solvent in which the compound represented by formula VII-3 is dissolved with silica gel to perform solid phase extraction of the compound represented by formula VII-3.

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