JP6605201B2 - Method for producing sugar amino acid derivative or glycopeptide derivative - Google Patents

Description

  The present invention relates to a method for producing a glycopeptide derivative or a sugar amino acid derivative. Specifically, by protecting all sugar hydroxyl groups with carbamate-based protecting groups that can be removed under acidic conditions, conventional sugar hydroxyl deprotection procedures that were separately required after the final deprotection of peptides and amino acids were performed. The present invention relates to a method for producing a glycopeptide derivative or a sugar amino acid derivative, which can be carried out simultaneously with the final deprotection of a peptide or amino acid.

  Since glycopeptide synthesis is mainly performed by solid phase synthesis, an acyl component is often used in excess. Therefore, it is necessary to protect the sugar hydroxyl group in order to prevent O-acylation to the sugar hydroxyl group.

  As a protecting group for the sugar hydroxyl group used in glycopeptide synthesis, acyl groups such as acetyl group and benzoyl group, benzyl protecting groups such as benzyl group, paramethoxybenzyl group and benzylidene group, and tert-butyl are generally used. Examples thereof include silyl groups such as dimethylsilyl group.

  In particular, acyl protecting groups and benzyl groups are widely used in the synthesis of sugar amino acids, which are raw materials for glycopeptides, because they are general-purpose protecting groups in sugar chain synthesis. However, since acyl-based protecting groups and benzyl groups are stable in trifluoroacetic acid (TFA) treatment, which is a deprotection condition for the side chain of a peptide chain and a condition for excision of a peptide chain from a solid phase, deprotection of the peptide chain portion Later, the sugar hydroxyl group needs to be deprotected. In the case of an acyl group, a free glycopeptide can be obtained by deprotecting by treatment with a base such as sodium hydroxide or sodium methoxide and the benzyl group by catalytic hydrogen reduction or trifluoromethanesulfonic acid (TfOH) -TFA treatment.

  However, it is known that side reactions such as racemization of peptide sites, by-product of dehydroalanine by β-elimination, and elimination of sugar residues occur under basic conditions, which are conditions for removing acyl groups (non-condensation). Patent Document 1). In the case of a benzyl group, if a peptide chain contains a sulfur-containing amino acid such as cysteine or methionine, it may become a catalyst poison of a catalyst for catalytic reduction (Non-patent Document 2). Furthermore, a side reaction in which the side chain of tyrosine is reduced by catalytic reduction has also been reported (Non-patent Document 3).

  In N-linked glycopeptide synthesis, a method has been reported in which a protecting group capable of deprotecting under acidic conditions is used as the protecting group for the sugar hydroxyl group, and the sugar hydroxyl group is deprotected simultaneously with the final deprotection of the peptide chain by TFA treatment.

  For example, there has been reported a method of simultaneously deprotecting a peptide chain and deprotecting a sugar hydroxyl group by protecting the hydroxyl group of GlcNAc with a TBDMS group and performing TFA treatment after solid-phase synthesis (Non-patent Document 4). However, in this method, since the Staudinger reaction using glycosyl azide cannot be used in the synthesis of sugar amino acid, it is necessary to synthesize the sugar amino acid by once reducing the azide group to an amino group and then condensing with the side chain carboxyl group of aspartic acid. is there. However, glycosylamine obtained by reducing the azide group is easily isomerized at the anomeric position, so that the resulting sugar amino acid is a mixture of the natural β-form and the non-natural alpha-form.

  In recent years, a method has been reported in which the Staudinger reaction can be used during sugar amino acid synthesis and the protective group of the sugar hydroxyl group is deprotected simultaneously with the final deprotection of the peptide chain by TFA treatment (Non-patent Document 5). However, this method requires nine steps when the synthesis of a sugar amino acid that is a raw material of a glycopeptide uses a commercially available glucosamine derivative as a starting material. Compared with the conventional method, in which the sugar chain amino acid synthesis with the acetyl group protecting the sugar hydroxyl group requires only two steps, the number of synthesis steps is greatly increased. Labor is required.

  In the synthesis of O-linked glycopeptides, it is common to use an acyl-type protecting group as a sugar-hydroxyl protecting group. As a reason, since an O-glycoside bond is generally an acetal bond, it may be decomposed under acidic conditions. Acyl-type protecting groups stabilize O-glycoside bonds even under TFA conditions, and are therefore widely used in glycopeptide synthesis. However, it is known that under basic conditions, which are the removal conditions, as described above, side reactions such as racemization of peptide sites, by-product of dehydroalanine due to β-elimination, and elimination of sugar residues occur. It has been.

  A benzyl group may be used as a hydroxyl-protecting group. However, deprotection by TfOH-TFA treatment may cause degradation of glycoside bonds due to strong acidity. Deprotection by catalytic reduction has problems such as modification of the amino acid site and catalyst poison as described above. In the case of a benzyl group or a silyl protecting group, the glycoside bond is decomposed by TFA treatment (Non-patent Document 6).

  Desirable protecting groups for sugar hydroxyl groups in glycopeptide synthesis are (1) easy introduction into sugar hydroxyl groups, (2) sugar amino acids as raw materials can be synthesized relatively easily, and (3) peptide sites by TFA treatment. It can be deprotected simultaneously with deprotection and does not affect the glycosidic bond. (4) It can be used for both N-linked and O-linked types.

Sjolin, P.M. Elofsson, M .; Kihlberg, J .; J. et al. Org Chem. 1996, 61, 560-565. Nakahara, Y. et al. Nakahara, Y .; Ito, Y .; Ogawa, T .; Tetrahedron Lett. 1997, 38, 7211. Guo, Z. W. Nakahara, Y .; Nakahara, Y .; Ogawa, T .; Carbohydr. Res. 1997, 303, 373. Holm, B.B. Linse, S .; and Kihlberg, J. et al. Tetrahedron 1998, 54, 11995. Y. Asahina et al. Org. Biomol. Chem. 2013, 11, 7199. Ning Shao et al. Org. Chem. 2003, l68, 9003.

  An object of the present invention is to use a sugar hydroxyl protecting group that is easy to introduce into a sugar hydroxyl group and synthesize a sugar amino acid, can be removed at the same time as a peptide protecting group by acid treatment, and does not affect the stability of the glycosidic bond. It was to provide an efficient method for producing glycopeptides.

  As a result of intensive studies on the above problems, the present inventors based on the new knowledge that glycopeptides can be efficiently produced by using carbamate-based protecting groups that can be removed under acidic conditions as protecting groups for sugar hydroxyl groups. The present invention has been completed.

That is, the present invention is the following production method and sugar derivative.
<1> A method for producing a sugar amino acid derivative or a glycopeptide derivative,
A production method comprising using, as a raw material, a sugar derivative or / and a sugar amino acid derivative in which all functional groups in a sugar residue are protected with a carbamate-based protecting group that can be removed under acidic conditions.

<2> The sugar derivative in which all functional groups in the sugar residue are protected with a carbamate-based protecting group that can be removed under acidic conditions is a sugar derivative represented by the following formula (1): The manufacturing method as described.
A-XY (1)
(In Formula (1), A is a monosaccharide or a sugar chain in which all functional groups are protected with a carbamate-based protecting group that can be removed under acidic conditions; Y is present or absent, and Y is absent. In the case, X is an azide group, a hydroxyl group, or a group having a leaving ability, and when Y is present, X is oxygen, sulfur, or selenium, and Y is an acyl group having 1 to 20 carbon atoms. A saturated or unsaturated hydrocarbon group or an aromatic group.)

<3> A sugar amino acid derivative in which all functional groups in the sugar residue are protected with a carbamate-based protecting group that can be removed under acidic conditions is a sugar amino acid derivative represented by the following formula (2) <1 > Or <2>.
A-Am (2)
(In Formula (2), A is a monosaccharide or a sugar chain in which all functional groups are protected with a carbamate-based protecting group that can be removed under acidic conditions; Am is an amino acid or an amino acid derivative; The group is protected with a free or protecting group or converted to an azide group, and the main chain carboxyl group is protected with a free or protecting group; the A-Am bond is linked to the side chain of Am and the reducing end anomer of A. It is a bond between carbon.)

<4> The production method according to <1> to <3>, wherein the carbamate-based protecting group that can be removed under acidic conditions is a tert-butoxycarbonyl (Boc) group.

（式（３）中、Ｙは存在するか存在せず、Ｙが存在しない場合は、Ｘはアジド基、水酸基、または脱離能を有する基であり、Ｙが存在する場合は、Ｘは酸素、硫黄、またはセレンであり、Ｙはアシル基、炭素数１から２０である飽和もしくは不飽和炭化水素基、または芳香族基であり； Ｚは水素、メチル基、またはｔｅｒｔ−ブチルオキシカルボニルオキシメチル基であり； Ｗは窒素、酸素、またはアジド基であり； Ｗがアジド基である場合は、Ｒ１とＲ２は存在せず、Ｗが酸素である場合は、Ｒ１はＢｏｃ基で、Ｒ２は存在せず、Ｗが窒素である場合は、Ｒ１はＢｏｃ基で、Ｒ２は水素、シリル基、アシル基、アルキル基、アリール基、またはアラルキル基である。） <5> A sugar derivative represented by the following formula (3).

(In Formula (3), Y is present or absent, and when Y is absent, X is an azide group, a hydroxyl group, or a group having a leaving ability, and when Y is present, X is oxygen. , Sulfur, or selenium, Y is an acyl group, a saturated or unsaturated hydrocarbon group having 1 to 20 carbon atoms, or an aromatic group; Z is hydrogen, a methyl group, or tert-butyloxycarbonyloxymethyl W is nitrogen, oxygen, or an azide group; when W is an azide group, R1 and R2 are not present; when W is oxygen, R1 is a Boc group and R2 is present And when W is nitrogen, R1 is a Boc group and R2 is hydrogen, a silyl group, an acyl group, an alkyl group, an aryl group, or an aralkyl group.)

（式（４）中、Ｘ１、Ｘ２はそれぞれ、水酸基をＢｏｃ基で保護したＬ−フコース残基、またはＢｏｃ基である。） <6> A sugar derivative represented by the following formula (4).

(In formula (4), X1 and X2 are each an L-fucose residue having a hydroxyl group protected with a Boc group, or a Boc group.)

  If the glycopeptide synthesis method using a carbamate-based protecting group that can be removed under acidic conditions of the present invention is used, it is very difficult to suppress by conventional methods. Glycopeptides can be synthesized without side reactions and side reactions such as elimination of sugar residues and cleavage of sugar chains. Furthermore, compared to peptide synthesis methods that use acyl-based protecting groups as sugar hydroxyl protecting groups, sugar hydroxyl group deprotection and peptide site deprotection can be performed simultaneously. We succeeded in improving the synthesis efficiency.

Hereinafter, the present invention will be described in detail.
The present invention relates to a method for producing a sugar amino acid derivative or a glycopeptide derivative, wherein all functional groups in a sugar residue are protected with a carbamate protecting group that can be removed under acidic conditions. It is a manufacturing method characterized by using a derivative as a raw material.

  The method for producing a sugar amino acid derivative of the present invention includes a case where the protected sugar derivative is condensed with an amino acid, or a case where the protected sugar derivative is condensed with the protected sugar amino acid derivative.

  In the method for producing a glycopeptide derivative of the present invention, when two or more protected sugar amino acid derivatives are condensed, or when the protected sugar derivative and a peptide are condensed, or the protected sugar amino acid derivative and an amino acid or When condensing with a peptide, when condensing the protected sugar derivative and two or more protected sugar amino acid derivatives, or condensing the protected sugar derivative with the protected sugar amino acid derivative and an amino acid or peptide. Includes cases.

A sugar amino acid as a raw material for the synthesis of a glycopeptide is synthesized by condensation of an amino acid and a sugar derivative. The sugar derivative used in the production method of the present invention is preferably a compound represented by the following formula (1).
A-XY (1)
(In Formula (1), A is a monosaccharide or sugar chain in which all functional groups are protected with a carbamate-based protecting group that can be removed under acidic conditions; Y is present or absent, and Y is present. If not, X is an azide group, hydroxyl group, or a group having a leaving ability such as a halogen group, an imidate group, an acetoxy group, and when Y is present, X is oxygen, sulfur or selenium; Is an acyl group, a saturated or unsaturated hydrocarbon group having 1 to 20 carbon atoms, or an aromatic group.)

The sugar amino acid derivative used in the production method of the present invention is preferably a compound represented by the following formula (2).
A-Am (2)
(In Formula (2), A is a monosaccharide or a sugar chain in which all functional groups are protected with a carbamate-based protecting group that can be removed under acidic conditions; Am is an amino acid or an amino acid derivative, and The main chain amino group is protected with a free or protective group or converted to an azide group, and the main chain carboxyl group is free or protected with a protective group; the A-Am bond is a reduction of Am side chain and A (This is a bond with the terminal anomeric carbon, and forms an amide bond, ester bond, ether bond, thioether bond, acetal bond, thioacetal bond, carbon-carbon bond, etc.)

  The type of carbamate-based protecting group is not particularly limited as long as it can be removed under acidic conditions. However, a Boc group, a 2- (trimethylsilyl) ethoxycarbonyl (Teoc) group, paramethoxybenzyloxy, which is easy to obtain and prepare an introduction reagent. Carbonyl (Z (OMe)) group, paramethylbenzyloxycarbonyl (Z (Me)) group, benzyloxycarbonyl (Z) group, isobornyloxycarbonyl (Iboc) group, 2- (3,5-dimethyloxyphenyl) ) -2-propyloxycarbonyl (Ddz) group, parabiphenylisopropyloxycarbonyl (Bpoc) group and 2-phenylisopropyloxycarbonyl group are preferable, and Boc group, Z (OMe) group and Teoc group are particularly preferable.

  The type of amino acid and amino acid derivative is not particularly limited, but is preferably an α-amino acid having a functional group in the side chain, and further, asparagine, which is an amino acid bonded to a sugar residue in a natural glycoprotein, Serine and threonine are more preferable. The steric structure of the amino acid is not particularly limited, and may be L-form, D-form, or a mixture of these enantiomers, but is preferably L-form.

  The main chain amino group and main chain carboxyl group of amino acids and amino acid derivatives may be free or protected with a protecting group. The type of protecting group of the main chain amino group is not particularly limited, but Boc group, Z group, Teoc group, 9-fluorenylmethoxycarbonyl (Fmoc) group, allyloxycarbonyl (Alloc) group commonly used in peptide synthesis, 2,2,2-trichloroethyloxycarbonyl (Troc) group is preferred, and Boc group and Fmoc group used in solid phase synthesis are particularly preferred. The main chain amino group may be converted to an azide group. The type of protecting group for the main chain carboxyl group is not particularly limited, but methyl ester, ethyl ester, phenacyl ester, benzyl ester, tert-butyl ester, trimethylsilylethyl ester, trimethylsilyl ester, trichloroethyl ester, paramethoxybenzyl ester, allyl Ester, 9-fluorenylmethyl ester is preferable, and in consideration of the removal of the carbamate-based protecting group protecting the sugar hydroxyl group under acidic conditions, it is removed under neutral conditions or weakly basic or weakly acidic conditions. Phenacyl ester, benzyl ester, trichloroethyl ester, allyl ester, and 9-fluorenylmethyl ester are preferred.

  The A-Am bond is a bond between the amino acid and the side chain of the amino acid derivative and the anomeric carbon at the reducing end of the sugar derivative, and the type of bond is not particularly limited as long as it is a covalent bond. Bonds, ether bonds, thioether bonds, acetal bonds, thioacetal bonds, and carbon-carbon are preferred, and in particular, amide bonds and acetal bonds, which are bonds between sugar residues and proteins, are formed in natural glycoproteins. Is preferred.

  The synthesis of sugar amino acids (A-Am bond formation) can be synthesized using techniques known to those skilled in the art. For example, N-linked sugar amino acid 3 can be synthesized from 2-deoxy-2-acetamido-β-D-glucopyranosyl azide (1) by the following method.

First, compound B is converted to tetra-Boc body 2 by allowing di-tert-butyl dicarbonate and N, N-dimethyl-4-aminopyridine to act on compound 1. Aspartic acid derivative [Fmoc-Asp (OPfp) -OBn] protected with a pentafluorophenyl group on the side chain carboxyl group, tributylphosphine, 3-hydroxy-4-oxo-3,4- By reacting with dihydro-1,2,3-benzotriazine (HOObt), compound 3 which is a sugar amino acid in which a sugar moiety and an amino acid moiety are amide-bonded can be prepared.

  O-linked sugar amino acid 15 can be synthesized by the following method.

By reacting Fmoc-threonine benzyl ester, N-iodosuccinimide and trifluoromethanesulfonic acid with 2-azidogalactose thioglycoside 14 in which the hydroxyl group is protected with a Boc group, a sugar having an acetal bond between the sugar moiety and the amino acid moiety Compound 15 which is an amino acid can be prepared.

  The glycopeptide can be synthesized using a sugar amino acid in which the main chain carboxyl group of Am in formula (2) is released or activated, using techniques known to those skilled in the art.

  The type of solvent that can be used for glycopeptide synthesis is not particularly limited, but in the case of chemical synthesis, an organic solvent is preferred. Examples of the organic solvent include methylene chloride, chloroform, dichloroethane, dimethylformamide, dimethylacetamide, tetrahydrofuran, ethyl acetate, 1-methyl-2-pyrrolidinone, acetonitrile, methanol, ethanol, or a combination of the above solvents. Preferred are methylene chloride, dimethylformamide, dimethylacetamide, and 1-methyl-2-pyrrolidinone.

  The reaction used for glycopeptide synthesis is not particularly limited as long as it is a reaction that forms an amide bond, but a condensing agent is preferably used. Examples of condensing agents include carbodiimide condensing agents (DCC, EDC / HCl, DIC), imidazole condensing agents (CDI, DMC), triazine condensing agents (DMT-MM), phosphonium condensing agents (BOP, PyBOP, PyCloP). PyBrop), uronium-based condensing agents (COMU, HBTU, HATU, TSTU) and the like. An additive may be added to these condensing agents. Examples of additives are 1-hydroxybenzotriazole (HOBt), 3,4-dihydro-3-hydroxy-4-oxo-1,2,3-benzotriazine (HOObt), 1-hydroxy-7-azabenzotriazole (HOAt), N-hydroxysuccinimide (HOSu), N-hydroxy-5-norbornene-2,3-dicarboximide (HONB), 2-hydroxyimino-2-cyanoacetic acid ethyl ester and the like.

  The present invention will be described in more detail below with reference to examples, but these examples show specific examples of the present invention and do not limit the present invention.

Synthesis of 3,4,6-Tri-O-tert-butoxycarbon-2-yl-2-deoxy-2- {N- (tert-butoxycarbonyl) -acetamido} -β-D-glucopyranosyl azide (2) (ex6-192)
2-Deoxy-2-acetamido-β-D-glucopyranosyl azide (1) (1.03 g, 4.17 mmol) and di-tert-butyl dicarbonate (5.1 g, 23.4) in tetrahydrofuran (60 mL). mmol) and N, N-dimethyl-4-aminopyridine (410 mg, 3.30 mmol) were added, and the mixture was stirred for 90 minutes under heating to reflux. After the solvent was distilled off under reduced pressure, the resulting residue was purified by silica gel column chromatography (developing solvent n-hexane: ethyl acetate = 6: 1) to obtain an amorphous solid compound 2 (2.68 g, quant.). Obtained.

Compound 2 ¹ H NMR (600 MHz, CDCl ₃ ): δ = 1.4-11.58 (m, 36H), 2.39 (s, 1.8H, conformer A), 2.44 (s, 1. 2H, conformer B), 3.82-3.85 (m, 0.4H, conformer B), 3.87-3.89 (m, 0.6H, conformer A), 4.15-4.19 ( m, 1.6H, conformer A and former B), 4.31-4.39 (m, 1H, former A and former B) 4.81-4.89 (m, 1.4H, former A and former B) ), 5.34 (d, 0.4H, J = 8.9 Hz, conformer B), 5.46 (dd, 0. 4H, J = 9.6 Hz, conformer B), 5.58 (dd, 0.6H, J = 10.3 Hz, conformer A), 5.65 (d, 0.6H, J = 8.9 Hz, conformer A)

^{N 2 -9-Fluorenylmethoxycarbonyl-N 4} - [3,4,6-Tri-O-tert-butoxycarbonyl-2-deoxy-2- {N- (tert-butoxycarbonyl) -acetamido} -β-D-glucopyranosyl] - L-asparagine tert-butyl ester
Compound 2 (93 mg, 0.14 mmol), Fmoc-Asp (OPfp) -OtBu (112 mg, 0.19 mmol) and 3,4-dihydro-3-hydroxy-4-oxo-1,2,3- Benzotriazine (33 mg, 0.20 mmol) was dissolved in a water / THF (2/98 v / v) mixed solution (1.6 mL) in an argon atmosphere, and then tributylphosphine (50 μL, 0.20). mmol) was added and stirred at room temperature for 4 hours. Ethyl acetate was added to the reaction solution, and the organic phase was washed with saturated brine and then dried over magnesium sulfate. After the solvent was distilled off under reduced pressure, the obtained residue was purified by silica gel column chromatography (developing solvent n-hexane: ethyl acetate = 3: 1) to obtain Compound 3 (148 mg, quant.).

Compound 3 ¹ H NMR (600 MHz, CDCl ₃ ): δ = 1.42-1.58 (m, 36H), 1.51 (s, 1.8H, conformer B), 1.60 (s, 7. 2H, conformer A), 2.33 (s, 0.6H, conformer B), 2.48 (s, 2.4H, conformer A), 2.64 (dd, 1H, J = 4.1 Hz, 16 .5 Hz), 2.84 (dd, 1H, J = 4.1 Hz, 16.5 Hz), 3.78-3.80 (m, 0.8 H, conformer A), 3.86-3. 89 (m, 0.2H, conformer B), 4.09-4.12 (m, 1H), 4.19-4.49 (m, 7H), 5.60 (dd, 0.8H, J = 9.6 Hz, conform r A), 5.74 (dd, 0.2H, J = 9.6 Hz, conformer B), 5.89 (dd, 1H, J = 9.6 Hz), 5.95 (d, 0.8H , J = 8.9 Hz, conformer A), 6.04-6.10 (m, 0.4H, conformer B), 6.15-6.17 (m, 1H), 7.29-7.32. (M, 4H), 7.39 (dd, 2H, J = 7.6 Hz), 7.60 (d, 2H, J = 7.6 Hz), 7.75 (d, 2H, J = 7. 6 Hz)

^{N 2 -9-Fluorenylmethoxycarbonyl-N 4} - (2-deoxy-2-acetamido-β-D-glucopyranosyl) -L-asparagine tert-butyl ester
Compound 3 (50.9 mg, 50 μmol) was dissolved in 80% TFA / methylene chloride solution (1 mL) and stirred at room temperature for 3 hours. Ether was added to the residue obtained by distilling off the solvent under reduced pressure, and the resulting powder was filtered to obtain Compound 4 (20.0 mg, 72%).

Compound 4 ¹ H NMR (600 MHz, DMSO-d6): δ = 1.74 (s, 3H), 2.42-2.51 (m, 1H), 2.57-2.61 (m, 1H) , 3.08-3.12 (m, 1H), 3.27-3.59 (m, 3H), 3.04-3.08 (m, 2H), 3.63-3.65 (m, 1H), 4.18-4.25 (m, 3H), 4.32-4.33 (m, 1H), 4.53 (t, 1H, J = 5.5 Hz), 4.79 (t , 1H, J = 9.6 Hz), 4.92 (d, 1H, J = 5.5 Hz), 4.97 (d, 1H, J = 4.8 Hz) 7.30 (t, 2H, J = 7.6 Hz), 7.38-7.43 (m, 3H), 7.69 (d, 2H, J = 7.6 Hz), 7.75 (brd, 1H, J 8.9 Hz), 7.87 (d, 2H, J = 7.6 Hz), 8.15 (brd, 1H, J = 8.9 Hz)

^{N 2 -9-Fluorenylmethoxycarbonyl-N 4} - [3,4,6-Tri-O-tert-butoxycarbonyl-2-deoxy-2- {N- (tert-butoxycarbonyl) -acetamido} -β-D-glucopyranosyl] - L-asparagine benzyl ester
Compound 2 (94 mg, 0.15 mmol), Fmoc-Asp (OPfp) -OBn (124 mg, 0.20 mmol) and 3,4-dihydro-3-hydroxy-4-oxo-1,2,3- Benzotriazine (35 mg, 0.22 mmol) was dissolved in a water / THF (2/98 v / v) mixed solution (1.6 mL) in an argon atmosphere, and then tributylphosphine (52 μL, 0.21). mmol) and stirred at room temperature for 2 hours. Ethyl acetate was added to the reaction solution, and the organic phase was washed with saturated brine and then dried over magnesium sulfate. After the solvent was distilled off under reduced pressure, the resulting residue was purified by silica gel column chromatography (developing solvent n-hexane: ethyl acetate = 4: 1) to obtain Compound 5 (160 mg, quant.).

Compound 5 ¹ H NMR (600 MHz, CDCl ₃ ): δ = 1.42-1.47 (m, 27H), 1.53 (s, 1.8H, conformer B), 1.60 (s, 7. 2H, conformer A), 2.33 (s, 0.6H, conformer B), 2.48 (s, 2.4H, conformer A), 2.64 (dd, 1H, J = 4.1 Hz, 16 .5 Hz), 2.84 (dd, 1H, J = 4.1 Hz, 16.5 Hz), 3.78-3.80 (m, 0.8 H, conformer A), 3.86-3. 89 (m, 0.2H, conformer B), 4.09-4.12 (m, 1H), 4.19-4.49 (m, 7H), 5.60 (dd, 0.8H, J = 9.6 Hz, conform r A), 5.74 (dd, 0.2H, J = 9.6 Hz, conformer B), 5.89 (dd, 1H, J = 9.6 Hz), 5.95 (d, 0.8H , J = 8.9 Hz, conformer A), 6.04-6.10 (m, 0.4H, conformer B), 6.15-6.17 (m, 1H), 7.29-7.32. (M, 4H), 7.39 (dd, 2H, J = 7.6 Hz), 7.60 (d, 2H, J = 7.6 Hz), 7.75 (d, 2H, J = 7. 6 Hz)

^{N 2 -9-Fluorenylmethoxycarbonyl-N 4} - [3,4,6-Tri-O-tert-butoxycarbonyl-2-deoxy-2- {N- (tert-butoxycarbonyl) -acetamido} -β-D-glucopyranosyl] - L-asparagine
Compound 5 (115 mg, 0.11 mmol) was dissolved in a mixed solvent of ethyl acetate (4 mL) and ethanol (2 mL), 10% palladium carbon (74 mg) was added, and hydrogen gas was bubbled at room temperature. Stir for 1 hour. Insoluble matter was removed by Celite filtration, and the solvent was distilled off under reduced pressure. The resulting residue was purified by silica gel column chromatography (developing solvent: chloroform: methanol = 30: 1) to give compound 6 (84 mg, 79%). Got.

Compound 6 ¹ H NMR (600 MHz, CDCl ₃ ): δ = 1.40-1.58 (m, 36H), 2.31 (s, 0.45H, conformer B), 2.47 (s, 2. 55H, conformer A), 2.68-2.88 (m, 2H), 3.80-3.88 (m, 1H), 4.08-4.13 (m, 1H), 4.21-4 .22 (m, 1H), 4.32-4.34 (m, 3H), 4.45-4.62 (brm, 1H), 4.82-4.88 (m, 2H), 5.58 (T, 0.85H, J = 9.6 Hz, conformer A), 5.74 (t, 0.15H, J = 9.6 Hz, conformer B), 5.92 (t, 0.85H, J = 9.6 Hz, conformer A), 6.10 (t, 0 15H, J = 9.6 Hz, conformer B),
6.17-6.31 (brm, 1H), 6.62 (brm, 1H), 7.30 (t, 1.7H, J = 6.9 Hz, conformer A), 7.38 (t, 1 .7H, J = 7.6 Hz, conformer A), 7.45 (t, 0.3H, J = 7.6 Hz, conformer B), 7.54 (t, 0.3H, J = 7.6) Hz, conformer B), 7.59 (d, 2H, J = 6.9 Hz), 7.74 (d, 1.7H, J = 7.6 Hz, conformer A), 7.81 (d, 0 .3H, J = 7.6 Hz, conformer B)

2,3,4-Tri-O-tert-butoxycarbonyl-α-L-fucopyranosyl- (1 → 6) -3,4-di-O-tert-butoxycarbonyl-2-deoxy-2- {N- (tert- butyoxycarbonyl) -acetamido} -β-D-glucopyranosyl azide (8)
Compound 7 (0.64 g, 1.64 mmol), di-tert-butyl dicarbonate (6.01 g, 27.6 mmol) and N, N-dimethyl-4-aminopyridine (160) were added to tetrahydrofuran (20 mL). mg, 1.31 mmol) was added, and the mixture was stirred for 60 minutes under heating to reflux. After the solvent was distilled off under reduced pressure, the resulting residue was purified by silica gel column chromatography (developing solvent n-hexane: ethyl acetate = 6: 1) to obtain amorphous solid compound 8 (1.44 g, 88%). Obtained.

Compound 11 ¹ H NMR (600 MHz, CDCl ₃ ): δ = 1.19-1.20 (m, 3H), 1.41-1.59 (m, 54H), 2.38 (s, 0.55H , Conformer A), 2.45 (s, 0.45H, conformer B), 3.65-3.75 (m, 2H), 3.83-3.89 (m, 1H), 4.13-4. .19 (m, 1.55H, conformer A and conformer B), 4.66-4.72 (m, 1H), 4.85 (dd, 0.45H, J = 8.9 Hz, conformer B), 4.90 (d, 0.45H, J = 4.1 Hz, conformer B), 4.92 (d, 0.55H, J = 4.1 Hz, conformer A), 5.06-35.09 ( m, 1H), 5.20-5.28 (m, 2.45H, conformer A and conformer B), 5.46-5.49 (dd, 0.45H, J = 8.9 Hz, conformer B), 5 .55-5.58 (m, 1.10H, conformer A)

^{N 2 -9-Fluorenylmethoxycarbonyl-N 4} - [2,3,4-Tri-O-tert-butoxycarbonyl-α-L-fucopyranosyl - (1 → 6) -3,4-di-O-tert-butoxycarbonyl-2 -Deoxy-2- {N- (tert-butoxycarbonyl) -acetamido} -β-D-glucopyranosyl] -L-asparagine benzyl ester (9) Compound 8 (299 mg, 0.30 mmol) and Fmoc-Asp OPfp) -OBn (257 mg, 0.42 mmol) and 3,4-dihydro-3-hydroxy-4-oxo-1,2,3-benzotriazine (74 mg, 0.45 mmol) in an argon atmosphere. After dissolving in water / THF (2/98 v / v) mixed solution (5 mL) in the atmosphere, tributylphosphine (104 μL, 0.42 mmol) was added and stirred at room temperature for 2 hours. Water and ethyl acetate were added to the reaction solution, and the organic phase was washed with water and saturated brine, and then dried over sodium sulfate. After the solvent was distilled off under reduced pressure, the obtained residue was purified by silica gel column chromatography (developing solvent n-hexane: ethyl acetate = 2: 1) to obtain Compound 9 (416 mg, 99%).

Compound 9 ¹ H NMR (600 MHz, CDCl ₃ ): δ = 1.19-1.20 (m, 3H), 1.41-1.47 (m, 45H), 1.59 (s, 7.65H , Conformer A), 1.71 (s, 1.35H, conformer B), 2.27 (s, 0.45H, conformer B), 2.46 (s, 2.55H, conformer A), 2.72. -2.77 (m, 1H), 2.88-2.95 (m, 1H), 3.62-3.67 (m, 1H), 3.74-3.76 (m, 1H), 3 .82-3.84 (m, 1H), 4.16-4.22 (m, 2.15H), 4.25-4.30 (m, 1H), 4.37-4.41 (m, 1H), 4.66-4.69 (m, 0.85H, conf rmer A), 4.73-4.80 (m, 2H), 4.94 (dd, 1H, J = 3.4, 10.3 Hz), 5.05-5.22 (m, 5.15H) ), 5.64 (dd, 0.85H, J = 8.9, 10.3 Hz, conformer A), 5.77 (t, 0.15H, J = 9.6 Hz, conformer B), 5. 88 (t, 0.85H, J = 9.6 Hz, conformer A), 6.01 (t, 0.15H, J = 8.9 Hz, conformer B), 6.22 (t, 1.7H, J = 7.6 Hz, conformer A), 6.33 (d, 0.15H, J = 8.9 Hz, conformer B), 7.27-7.30 (m, 7H), 7.38 (t , 2H, J = 7.6 Hz), 7.59 ( , 2H, J = 7.6 Hz), 7.74 (d, 2H, J = 7.6 Hz)

^{N 2 -9-Fluorenylmethoxycarbonyl-N 4} - [2,3,4-Tri-O-tert-butoxycarbonyl-α-L-fucopyranosyl - (1 → 6) -3,4-di-O-tert-butoxycarbonyl-2 Synthesis of -deoxy-2- {N- (tert-butoxycarbonyl) -acetamido} -β-D-glucopyranosyl] -L-asparagine (10) Compound 9 (197 mg, 0.14 mmol) in THF (10 mL) The mixture was dissolved, palladium carbon-ethylenediamine complex (120 mg) was added, and the mixture was stirred at room temperature for 3 hours while bubbling hydrogen gas. Insoluble matter was removed by Celite filtration, and the solvent was distilled off under reduced pressure. The resulting residue was purified by silica gel column chromatography (developing solvent chloroform: methanol = 9: 1) to give compound 10 (170 mg, 93%). Got.

Compound 10 ¹ H NMR (600 MHz, CDCl ₃ ): δ = 1.20 (d, 3H, J = 6.2 Hz), 1.41-1.48 (m, 45H), 1.52 (s, 1.8H, conformer B), 1.59 (s, 7.2H, conformer A), 2.32 (s, 0.6H, conformer B), 2.47 (s, 2.4H, conformer A), 2.70-2.89 (m, 2H), 3.70-3.73 (m, 2H), 3.84-3.87 (m, 1H), 4.18-4.24 (m, 2H) ), 4.32-4.42 (brm, 2H), 4.59 (brm, 1H), 4.70 (t, 1H, J = 9.6 Hz), 4.79 (t, 1H, J = 9.6 Hz), 4.92 (dd, 1H, J = 3.4, 1 0.3 Hz), 5.09-5.27 (m, 3H), 5.64 (t, 0.8H, J = 9.6 Hz, conformer A), 5.74 (t, 0.2H, J = 9.6 Hz, conformer B), 5.87 (t, 0.8H, J = 9.6 Hz, conformer A), 6.13 (brd, 0.8H, J = 6.2 Hz, conformer) A), 6.25 (br, 0.2H, conformer B), 6.51 (br, 0.8H, conformer A), 7.26-7.33 (m, 2H), 7.38-7. 40 (m, 2H), 7.60 (d, 2H, J = 7.6 Hz), 7.75 (d, 2H, J = 7.6 Hz)

^{N 2 -9-Fluorenylmethoxycarbonyl-N 4} - [2,3,4-Tri-O-tert-butoxycarbonyl-α-L-fucopyranosyl - (1 → 6) -3,4-di-O-tert-butoxycarbonyl-2 -Deoxy-2- {N- (tert-butyoxycarbonyl) -acetamido} -β-D-glucopyranosyl] -L-asparagine tert-butyl ester (11)
Compound 8 (107 mg, 0.11 mmol), Fmoc-Asp (OPfp) -OtBu (86 mg, 0.15 mmol) and 3,4-dihydro-3-hydroxy-4-oxo-1,2,3- Benzotriazine (25 mg, 0.15 mmol) was dissolved in a water / THF (2/98 v / v) mixed solution (3 mL) in an argon atmosphere, and then tributylphosphine (36 μL, 0.15 mmol). And stirred at room temperature for 2 hours. Water and ethyl acetate were added to the reaction solution, and the organic phase was washed with water and saturated brine, and then dried over sodium sulfate. After evaporating the solvent under reduced pressure, the resulting residue was purified by silica gel column chromatography (developing solvent n-hexane: ethyl acetate = 2: 1) to obtain Compound 11 (135 mg, 92%).

Compound 11 ¹ H NMR (600 MHz, CDCl ₃ ): δ = 1.18-1.19 (m, 3H), 1.41-1.46 (m, 54H), 1.60 (s, 7.5H , Conformer A), 1.75 (s, 1.5H, conformer B), 2.31 (s, 0.5H, conformer B), 2.48 (s, 2.5H, conformer A), 2.68. -2.71 (m, 1H), 2.81-2.87 (m, 1H), 3.61-3.65 (m, 1H), 3.76-3.78 (m, 1H), 3 .82-3.83 (m, 1H), 4.18-4.32 (m, 3H), 4.38-4.43 (m, 1H), 4.50-4.51 (m, 1H) , 4.75-4.83 (m, 2H), 4.93-4.95 (m, H), 5.09-5.12 (m, 2H), 5.19 (m, 1H), 5.66 (t, 0.83H, J = 9.6 Hz), 5.74 (t, 0 .17H, J = 9.6 Hz), 5.86 (t, 0.83H, J = 9.6 Hz), 6.02 (t, 0.17H, J = 9.6 Hz), 6.06 (D, 0.83H, J = 9.6 Hz), 6.17 (d, 0.17H, J = 9.6 Hz), 6.27-6.31 (m, 1H), 7.29- 7.32 (m, 2H), 7.39 (t, 2H, J = 7.6 Hz), 7.61-7.63 (m, 2H), 7.75 (d, 2H, J = 7. 6 Hz)

^{N 2 -9-Fluorenylmethoxycarbonyl-N 4} - [α-L-fucopyranosyl - (1 → 6) -2-deoxy-2-acetamido-β-D-glucopyranosyl] -L-asparagine tert-butyl ester (12)
Compound 11 (107 mg, 77 μmol) was dissolved in a 20% trifluoroacetic acid / dichloromethane solution (5 mL) at 0 ° C. and stirred for 4 hours. Toluene (10 mL) was added to the reaction solution, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (developing solvent: chloroform: methanol = 3: 1) to obtain Compound 12 (51 mg, 91%).

Compound 12 ¹ H NMR (600 MHz, DMSO-d6): δ = 1.05 (d, 3H, J = 6.9 Hz), 1.76 (s, 3H), 2.45 (dd, 1H, J = 7.56, 15.8 Hz), 2.59 (dd, 1H, J = 5.5, 15.8 Hz), 3.08-3.12 (m, 1H), 3.27-3. 59 (m, 3H), 3.75 (d, 1H, J = 10.31 Hz), 3.85 (q, 1H, J = 6.9 Hz), 4.18-4.32 (m, 6H) ), 4.44 (bs, 1H,), 4.61 (d, 1H, J = 2.8 Hz), 4.80 (t, 1H, J = 8.9 Hz), 4.99 (d, 1H, J = 4.8 Hz), 5.07 (d, 1H, J = 4.8 Hz), 7.31 (t, 2H, J = 7.6 Hz), 7. 0t, 2H, J = 7.6 Hz), 7.70 (dd, 2H, J = 2.8, 6.87 Hz), 7.78 (d, 1H, J = 8.9 Hz), 7. 87 (d, 2H, J = 7.6 Hz), 8.15 (d, 1H, J = 8.9 Hz)

To tetrahydrofuran (3 mL) was added compound 13 (118 mg, 0.32 mmol), di-tert-butyl dicarbonate (419 mg, 1.92 mmol) and N, N-dimethyl-4-aminopyridine (23 mg, 0 .19 mmol) was added, and the mixture was stirred for 120 minutes under reflux. After the solvent was distilled off under reduced pressure, the resulting residue was purified by silica gel column chromatography (developing solvent n-hexane: ethyl acetate = 8: 1) to obtain Compound 14 (243 mg, quant.).

Compound 18 ¹ H NMR (600 MHz, CDCl ₃ ): δ = 1.32 (s, 9H), 1.46-1.48 (m, 27H), 2.44 (s, 3H), 3.72 ( dd, 1H, J = 10.3 Hz), 3.82-3.85 (m, 1H), 4.16-4.23 (m, 2H), 4.44 (d, 1H, J = 10. 3 Hz), 4.59 (dd, 1H, J = 3.4 Hz, 10.3 Hz), 5.25 (dd, 15H, J = 3.4 Hz), 7.16 (d, 1H, J = 8. 3 Hz), 7.25 (dd, 1H, J = 8.3 Hz, 10.3 Hz), 7.68 (d, 1H, J = 2.1 Hz)

Compound 14 (71 mg, 106 μmol) and Fmoc-Thr-OBn (31 mg, 71 μmol) were dissolved in dichloromethane (2 mL), molecular sieve 4A (200 mg) was added, and the mixture was cooled to −40 ° C. . N-iodosuccinimide (48 mg, 213 μmol) and trifluoromethanesulfonic acid (2.8 μL, 32 μmol) were added, and the mixture was stirred at −40 ° C. for 2 hours. The reaction solution was filtered through celite, washed with ethyl acetate, and the organic phase was washed with 5% aqueous sodium thiosulfate solution, saturated aqueous sodium hydrogen carbonate solution and saturated brine, and then dried over sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (developing solvent n-hexane: ethyl acetate = 3: 1) to obtain compound 15 as α form (20.2 mg, 31%). Obtained as a mixture of β-form (15.6 mg, 24%).

Compound 15α form ¹ H NMR (600 MHz, CDCl ₃ ): δ = 1.32 (d, 3H, J = 6.9 Hz), 1.45 (s, 9H), 1.50 (s, 9H), 1.51 (s, 9H), 3.63 (dd, 1H, J = 3.4 Hz, 11.0 Hz), 4.09-4.15 (m, 2H), 4.17-4.22 (M, 1H), 4.26 (dd, 1H, J = 7.6 Hz), 4.34 (dd, 1H, J = 7.6, 10.3 Hz), 4.38-4.48 ( m, 3H), 4.86 (d, 1H, J = 4.12 Hz), 5.02 (dd, 1H, J = 3.4, 11.0 Hz), 5.17 (d, 1H, J = 11.7 Hz), 5.25 (d, 1H, J = 11.7 Hz), 5.34 (dd, 1H, J = 2.8 Hz), 5.6 (D, 1H, J = 9.6 Hz), 7.29-7.41 (m, 9H), 7.63 (d. 2H, J = 6.9 Hz), 7.76 (d, 2H, J = 7.6 Hz)

Compound 15β-form ¹ H NMR (600 MHz, CDCl ₃ ): δ = 1.33 (d, 3H, J = 6.2 Hz), 1.46 (s, 9H), 1.50 (s, 9H), 1.51 (s, 9H), 3.68-3.71 (m, 2H), 4.08-4.19 (m, 2H), 4.25 (dd, 1H, J = 7.6 Hz) 4.33-4.39 (m, 3H), 4.40 (dd, 1H, J = 3.4 Hz, 11.0 Hz), 4.90 (dd, 1H, J = 2.1 Hz, 9.6 Hz), 4.61-4.64 (m, 1H), 5.16-5.19 (m, 2H), 5.27 (d, 1H, J = 12.3 Hz), 5. 63 (d, 1H, J = 9.6 Hz), 7.29-7.34 (m, 3H), 7.35-7.40 (m, 6H), 7.61 dd, 2H, J = 8.3 Hz), 7.75 (d, 2H, J = 7.6 Hz)

Synthesis of dipeptide Compound 10 (170 mg, 0.13 mmol); Glycine tert-butyl hydrochloride (22 mg, 0.13 mmol) and 1-hydroxybenzotriazole monohydrate (24 mg, 0.16 mmol) Was dissolved in DMF (3 mL), and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (30 mg, 0.16 mmol) and N, N-diisopropylethylamine (46 μL, 0.0. 26 mmol) was added and stirred at room temperature for 2 hours. Ethyl acetate was added to the reaction solution, and the organic phase was washed with 5% aqueous citric acid solution, aqueous saturated sodium hydrogen carbonate aqueous solution, water and saturated brine, and then dried over sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (developing solvent n-hexane: ethyl acetate = 1: 1) to obtain Compound 16 (167 mg, 91%). The obtained compound 16 (167 mg, 0.12 mmol) was dissolved in a 20% trifluoroacetic acid / dichloromethane solution at 0 ° C. and stirred at 0 ° C. for 4 hours. Diethyl ether and 20% aqueous acetonitrile were added to the solution, the aqueous phase was separated and lyophilized. 56 mg of the obtained residue (85 mg) was purified by HPLC to obtain Compound 17 (19 mg, 32%).

The production method and sugar derivative of the present invention can be used for the production of a sugar amino acid derivative or a glycopeptide derivative.

Claims (6)

A method for producing a sugar amino acid derivative or a glycopeptide derivative, comprising:
Using as a raw material a sugar derivative or / and a sugar amino acid derivative in which all functional groups in the sugar residue are protected with a carbamate-based protecting group that can be removed under acidic conditions ; and
Carbamate-based protecting groups that can be removed under acidic conditions in which all functional groups in the sugar residue are protected, and protecting groups in amino acid residues or peptide residues are simultaneously deprotected under acidic conditions. The manufacturing method characterized by including a process.
The production according to claim 1, wherein the sugar derivative in which all functional groups in the sugar residue are protected with a carbamate-based protecting group that can be removed under acidic conditions is a sugar derivative represented by the following formula (1). Method.
A-XY (1)
(In Formula (1), A is a monosaccharide or a sugar chain in which all functional groups are protected with a carbamate-based protecting group that can be removed under acidic conditions; Y is present or absent, and Y is absent. In the case, X is an azide group, a hydroxyl group, or a group having a leaving ability, and when Y is present, X is oxygen, sulfur, or selenium, and Y is an acyl group having 1 to 20 carbon atoms. A saturated or unsaturated hydrocarbon group or an aromatic group.) The sugar amino acid derivative obtained by protecting all functional groups in the sugar residue with a carbamate-based protecting group that can be removed under acidic conditions is a sugar amino acid derivative represented by the following formula (2): The manufacturing method as described in.
A-Am (2)
(In Formula (2), A is a monosaccharide or a sugar chain in which all functional groups are protected with a carbamate-based protecting group that can be removed under acidic conditions; Am is an amino acid or an amino acid derivative; The group is protected with a free or protecting group or converted to an azide group, and the main chain carboxyl group is free or protected with a protecting group; the A-Am bond is the anomeric side chain of Am and the reducing end of A It is a bond between carbon.)   The production method according to claim 1, wherein the carbamate-based protecting group that can be removed under acidic conditions is a tert-butoxycarbonyl (Boc) group. A sugar derivative represented by the following formula (3).

(In Formula (3), Y is present or absent, and when Y is absent, X is an azide group, hydroxyl group, halogen group, imidate group or acetoxy group , and when Y is present, X is Oxygen, sulfur, or selenium, Y is an acyl group, a saturated or unsaturated hydrocarbon group having 1 to 20 carbon atoms, or an aromatic group; Z is hydrogen, a methyl group, or tert-butyloxycarbonyloxy a methyl group; W is nitrogen Motoma other is an azido group; when W is an azido group, R1 and R2 is not present, when W is nitrogen, R1 is Boc group, R2 is Hydrogen, silyl group, acyl group, alkyl group, aryl group, or aralkyl group.)
A sugar derivative represented by the following formula (4).

(In formula (4), X1 and X2 are each an L-fucose residue having a hydroxyl group protected with a Boc group, or a Boc group.)