JPH085914B2 - Glycosyl-protein derivative - Google Patents

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a glycosyl-protein derivative and an intermediate thereof which are useful as a carrier for selectively delivering a drug to a target tissue, particularly bone marrow or brain.

[0002]

Bone marrow tissue is composed of red blood cells, lymphocytes, monocytes,
It is a tissue that produces granulocytes, and is the most important tissue responsible for hematopoiesis and immunity in the body. Radiation therapy and anticancer drug administration in cancer patients cause the decrease of bone marrow function.
It is an unavoidable side effect at present, and as a result of myelosuppression, a serious decrease in white blood cells and platelets, especially a severe infection associated with a decrease in granulocytes has become a serious problem.

Further, in order for the drug to be transported to the brain tissue, it has to pass through the blood-brain barrier, which impedes the migration of the drug into the brain. Therefore, there is a demand for the development of means for delivering a drug into the brain.

On the other hand, with the recent development of immunology, many physiologically active proteins have been isolated and purified, and their pharmacological actions have been clarified. Further, with the development of genetic engineering technology, many cytokines, for example, erythropoietin which is a hematopoietic hormone of erythrocytes, several kinds of colony stimulating factor (CSF) which is a hematopoietic factor of leukocytes, and α-,
β-, γ-interferon, interleukin 2 and the like have become possible to be mass-produced, and their application to pharmaceuticals has been attempted.

However, many cytokines
After being administered in vivo, it is rapidly metabolized,
There is a problem that the manifestation of drug efficacy in the target tissue cannot be achieved sufficiently.

In order to solve this problem, antibodies, glycoproteins, lipoproteins, lectins, hormones, liposomes, deoxyribonucleic acids, polysaccharides, synthetic polymers, as carriers for delivering physiologically active substances and synthetic drugs to target sites, A wide range of substances have been proposed, including naturally occurring polymers such as polyamino acids, molecular assemblies and synthetic polymers.

For example, Proc. Nath. Acad. Sci. USA 84,
1487-1491 (1987) reported a study in which some cytokines were modified with a synthetic polymer such as polyethylene glycol to prolong the half-life in vivo without losing the activity of the cytokines themselves.

Further, in Japanese Patent Laid-Open No. 63-152393, a polyethylene glycol derivative having a sugar chain can be used for modification of cytokines, and this modified protein delays clearance in a living body or can be used in specific cells. -It has been suggested to be used to improve delivery to tissues. However, there are no reports of bone marrow or brain tropism.

[0009]

The present invention utilizes sugar recognition on the surface of bone marrow or brain cells to deliver more drugs (cytokines and low molecular weight pharmaceuticals) to the target tissue, bone marrow or brain tissue. It is an invention of a drug delivery carrier that makes it possible to delay the in vivo half-life of a drug, and develops a system (Targeting Drug Delivery System) for delivering a target drug to a target tissue. It is intended.

In particular, the present invention relates to patients with hematopoietic function diseases such as aplastic anemia and bone marrow dysfunction such as immunodeficiency associated with various lymphocyte diseases, as well as myeloid leukemia, myeloma and plasma cells. For patients with myelogenous cancer such as tumors and multiple myeloma and patients who have decreased bone marrow function due to side effects associated with cancer treatment, etc., by efficiently collecting drugs used for the treatment in bone marrow tissues, Can delay the in vivo half-life of the drug,
The purpose is to develop a drug delivery carrier that can be expected to have higher efficacy.

Further, the present invention aims at the development of a carrier for drug delivery directly into the brain for the treatment of Alzheimer's disease and the like.
It enables treatment while maintaining its activity and reducing the possibility of side effects.

[0012]

The present invention SUMMARY OF] has the formula: _{[R-O- (CH 2) n} -CO- ] _m -Z (I) (wherein R represents a glycosyl group, Z is expressed protein , N represents 3 to 14, and m represents 10 to 50), which is a drug delivery carrier containing the glycosyl-protein derivative.
Particularly, those having m of 10 to 50 are preferable as carriers for delivery to bone marrow, and those having m of 20 to 50 are preferable as carriers for delivery to the brain.

In addition, the formula: [ROC ₆ H ₄ —NH-CS-] _m -Z (II) (wherein R represents a glycosyl group, Z represents a protein, and m represents 5 to 50). Represents a glycosyl-protein derivative.

[0014] wherein: _{[R-O- (CH 2) 2} O (CH 2) 2 O (CH 2) 3 -CO- ] _m -Z (III) (wherein R represents a glycosyl group, Z Represents a protein and m represents 5 to 50), and is a glycosyl-protein derivative.

The compound of the present invention can be used as a drug delivery carrier for target cells, organs, organs, particularly bone marrow or brain tissue. The drug carrier has (i) good biocompatibility, (ii) is stable for a certain period of time after administration, and (iii) is released by chemical or enzymatic reaction when the drug reaches its site of action. Is required.

The sugar chain constituting the carrier of the above formula can be used as a target identification site having the ability to specifically recognize a target cell organ, organ or the like. Examples of the glycosyl group represented by R include a xylopyranosyl group, a mannopyranosyl group, a fucopyranosyl group, a 2-galactopyranosyl group, a 2-acetamido-2-deoxy-fucopyranosyl group, a 2-acetamido-2-deoxy-mannopyranosyl group, and a 2-acetamido-2-deoxy-mannopyranosyl group. Monosaccharides such as acetamido-2-deoxy-galactopyranosyl group; mannopyranosyl-mannopyranosyl group, (2-acetamido-2-deoxy-mannopyranosyl) -mannopyranosyl group, (2-acetamido-2-deoxy-glucopyranosyl) -mannopyranosyl Group, fucopyranosyl- (2-acetamido-2-deoxy-glucopyranosyl) group, galactopyranosyl- (2-acetamido-2-deoxy-glucopyranosyl) group, galactopyranosyl- (2-acetamido-2-) Oxy - mannopyranosyl) group, galactopyranosyl - disaccharide or di like glucopyranosyl group (2-acetamido-2-deoxy - glucopyranosyl) - mannopyranosyl group, di (galactopyranosyl)
Mention may be made of trisaccharides such as the 2-acetamido-2-deoxy-glucopyranosyl group. Particularly preferably, a mannopyranosyl group, a fucopyranosyl group, a 2-acetamido-2-deoxy-fucopyranosyl group, a mannopyranosyl-mannopyranosyl group, a (2-acetamido-2-deoxy-glucopyranosyl) -mannopyranosyl group, and a galactopyranosyl-glucopyranosyl group are can give.

In the compound of the formula (I), the carboxylic acid as the modifier that connects the sugar chain and the protein has 3 to 14 alkylene moieties, preferably octanoic acid. The bond between the sugar chain and the carboxylic acid may be an α-bond or a β-bond. Examples of the Z protein include, in addition to proteins such as human serum albumin, cytokines interleukins, erythropoietin, interferon, which are physiologically active proteins themselves.
It may be a tissue plasminogen activator, ulcer necrosis factor (TNF) or colony stimulating factor (CSF). Although these proteins themselves do not have organ specificity, they become target-identifying by being chemically modified with sugar chains capable of target-identifying.

To prepare the carrier of formula (I), for example, an alkyl ester of glycosyl-alkanoic acid or 2-
Acetamide-2-deoxy-glycosyl-alkanoic acid alkyl ester is reacted with hydrazine, the obtained acid hydrazide is converted into an acid azide by a conventional method, and this is reacted with protein to react glycosyl-alkanoic acid with amino acid in protein. The desired product is obtained by amide bond with a part of the group. The number of modifying groups bonded is 5 to 50 mol per protein molecule. The degree of modification is
It can be selected by increasing or decreasing the molar ratio of glycosyl-alkanoic acid to protein or by increasing or decreasing the reaction solution concentration of protein and glycosyl-alkanoic acid.

The solvent used in the reaction may be any solvent as long as it does not interfere with the reaction, and examples thereof include phosphate buffer, Tris buffer, acetate buffer, borate buffer and the like. The reaction is carried out near neutrality at 0 to room temperature. The reaction solution is purified by a usual protein purification method such as dialysis, ion exchange chromatography, gel filtration, etc. to obtain the desired product. The number of modifying groups introduced can be known by hydrolyzing the modified protein with hydrochloric acid and then measuring by the Elson-Morgan method or the like.

The reaction of glycosyl-alkanoic acid with protein can also be produced by reacting in the presence of a condensing agent such as a water-soluble carbodiimide. Further, other active derivatives of alkanoic acid may be used, and examples of such active derivatives include amide compounds, active esters, active thioesters and the like. These active derivatives can be appropriately selected by those skilled in the art in order to form an amide bond with a protein.

The intermediate is a glycosyl-carboxylic acid of formula (IV) or its active derivative. _{R-O- (CH 2) n} -COOH (IV) herein, R represents particularly xylopyranosyl group, fucopyranosyl group, galactopyranosyl group, 2-acetamido-2-
Deoxy-fucopyranosyl group, 2-acetamido-2-
Deoxy-mannopyranosyl group, 2-acetamido-2
-Deoxy-galactopyranosyl group, (2-acetamido-2-deoxy-mannopyranosyl) -mannopyranosyl group, (2-acetamido-2-deoxy-glucopyranosyl) -mannopyranosyl group, fucopyranosyl-
(2-acetamido-2-deoxy-glucopyranosyl) group, galactopyranosyl- (2-acetamido-2)
-Deoxy-glucopyranosyl) group, galactopyranosyl- (2-acetamido-2-deoxy-mannopyranosyl) group, galactopyranosyl-glucopyranosyl group,
A glycosyl group such as a di (2-acetamido-2-deoxy-glucopyranosyl) -mannopyranosyl group or a di (galactopyranosyl) -2-acetamido-2-deoxy-glucopyranosyl group.

To prepare the intermediate of formula (IV), (A) a hydroxyl group-protected glycosyl halide is reacted with a ω-hydroxyalkanoic acid alkyl ester to give a glycosyl-α (or β) -alkanoic acid alkyl ester. (B) Alternatively, the 2-acetamido-2-deoxy-glycosyl-alkanoic acid alkyl ester can be obtained by deprotecting it as an ester. Can be obtained. Further, if necessary, it can be led to another active derivative.

Specific methods for preparing the compounds of formula (I) and formula (IV) will become apparent by referring to the following reaction scheme.

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The carrier of the formula (II) can be produced from the intermediate of the formula: R—O—C ₆ H ₄ —NH ₂ (V) by the method shown in the following reaction scheme. .

[0041]

[Chemical 17]

To prepare a carrier of formula (III), the following intermediates of the formula: R--O-(CH ₂ ) ₂ O (CH ₂ ) ₂ O (CH ₂ ) ₃ --COOH (VI) It can be produced by the method shown in the reaction scheme.

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[0047]

INDUSTRIAL APPLICABILITY The glycosyl-protein derivative of the present invention binds a drug as shown in Reference Example 56 below, and when this is administered to animals as shown in Test Examples, the drug is concentrated in bone marrow or brain tissue. Can be distributed in

[0048]

EXAMPLES Examples, reference examples and test examples of the present invention will be shown below.

Reference Example 1 6-Methoxycarbonylhexanol (2) Pimelic acid monomethyl ester (1), 25 g (156)
Mmol) and 21.7 ml of triethylamine (156
100 ml of anhydrous tetrahydrofuran of
Cool to −18 ° C. under nitrogen stream. A solution of 15 ml (156 mmol) of ethyl chlorocarbonate in 100 ml of anhydrous tetrahydrofuran was added dropwise to this solution while stirring. After stirring for 20 minutes at the same temperature, the precipitated salt was filtered off and washed with a small amount of anhydrous tetrahydrofuran. The filtrate and washings were combined and ice-cooled sodium borohydride 14.7 g (39
(0 mmol) in 150 ml of water. After further stirring at the same temperature for 1 hour, the mixture was partitioned into ethyl acetate and an aqueous layer.
The organic layer was washed with water, dried over magnesium sulfate and then concentrated to dryness under reduced pressure to obtain 18 g of a syrup-like residue. The residue was purified by silica gel column chromatography (hexane: ethyl acetate = 2: 1) to obtain 3.45 g of compound (2).

IR (film) cm ⁻¹ : 3400, 1737, 1438, 1207, 1174 ¹ H-NMR (CDCl ₃ , δppm): 3.68 (3H, s, OMe), 3.66 (2H, t, J = 6.4Hz _{, HOCH 2 -), 2.34 (} 2H, t, J = 7.2Hz, -CH 2 CO-), 1.75~ 1.30 (8H, m, - (CH 2) 4 -)

Elemental analysis C ₇ H ₁₄ O ₃ Calculated value C 57.51% H 9.65% Measured value C 57.64% H 9.82%

Reference Example 2 6-Methoxycarbonylhexyl 2,3,4,6-tetra-O-benzyl-β-D-mannopyranoside (5)
And 6-methoxycarbonylhexyl 2,3,4
6-Tetra-O-benzyl-α-D-mannopyranoside (6)

2,3,4,6-tetra-O-benzyl-
α-D-mannopyranosyl chloride (3), 608 mg
(1.087 mmol) and 6-methoxycarbonylhexanol (2) 159 mg (1.087 mmol) in dry toluene solution, molecular sieve 4A 750 mg.
Was added, and the mixture was stirred at room temperature for 30 minutes under a nitrogen stream. Then, 500 mg of silver silicate was added and stirred for 72 hours.
The reaction mixture was filtered through Celite, and the filtrate was concentrated to dryness under reduced pressure to give 720 mg of a syrupy residue. The residue was purified using a Rover column (hexane: ethyl acetate = 4: 1) to give compound (5) 380 mg (yield 52.2%).
And 130 mg of compound (6) (yield 17.9%) were obtained as syrupy substances.

Compound (5) [α] ^22.50 _D -47.1 ° (C 0.550, CHCl ₃ ) IR (film) cm ⁻¹ : 1740, 1440, 1365, 1105 ¹ H-NMR (CDCl ₃ , δppm): 7.5 〜 7.1 (20H, m, ph), 4.36 (1H, s, H-1), 3.65 (3H, s, OMe), 2.31 (2H, t, J = 7.5Hz, -CH ₂ CO-) 1.7 ~ 1.4 ( _{8H, m, - (CH 2} ) 4 -)

Elemental analysis C ₄₁ H ₆₈ O ₈ Calculated value C 73.62% H 7.23% Measured value C 73.25% H 7.04%

Compound (6) [α] ^22.50 _D +21.1 ° (C 1.05, CHCl ₃ ) IR (film) cm ⁻¹ : 1738, 1455, 1275, 1105 ¹ H-NMR (CDCl ₃ , δppm): 7.4 〜 7.2 (20H, m, ph), 4.62 (1H, s, H-1), 3.64 (3H, s, OMe), 2.29 (2H, t, J = 7.5Hz, -CH ₂ CO-) 1.8 ~ 1.2 ( _{12H, m, - (CH 2} ) 4 -)

Elemental analysis C ₄₁ H ₆₈ O ₈ Calculated value C 73.62% H 7.23% Measured value C 73.59% H 7.11%

Reference Example 3 8-Methoxycarbonyloctyl 2,3,4,6-tetra-O-benzyl-β-D-mannopyranoside (7)
And 8-methoxycarbonyloctyl 2,3,4,
6-Tetra-O-benzyl-α-D-mannopyranoside (8)

Bull. Chem. Soc. Jpn., 49, 2639 (1976)
2,3,4,6-tetra-O- synthesized by the method described in 1.
Benzyl-α / β-D-mannopyranose 1.0 g
Anhydrous tetrahydrofuran solution 2 of (1.85 mmol)
90 ml (1.87 mmol) of an oil suspension of 50% sodium hydride was added to 0 ml under nitrogen flow while cooling with ice,
The mixture was stirred at the same temperature for 40 minutes. Then, 8-methoxycarbonyloctyltrifluoromethanesulfonic acid ester 6
4 ml of a solution of 00 mg (1.87 mmol) in anhydrous tetrahydrofuran was added dropwise, and the mixture was further stirred for 2 hours. After pouring the reaction solution into ice water, the product was extracted with ethyl acetate. The ethyl acetate phase was washed with water, dried over magnesium sulfate, and then the solvent was distilled off under reduced pressure to obtain 1.3 g of a syrup-like residue. The residue was purified using a Rover column (hexane: ethyl acetate = 6: 1) to obtain 650 mg of compound (7) (yield 56.7%) and 171 mg of compound (8) (yield 14.9%). .

Compound (7) mp 54.5 to 55 ° C. [α] ²⁵ _D -46.7 ° (C 0.85, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 1732, 1497, 1454, 1362, 1100 ¹ H-NMR ( CDCl ₃ , δppm): 7.6 ~ 7.1 (20H, m, ph), 4.61 (1H, s, H-1), 3.66 (3H, s, OMe), 2.30 (2H, t, J = 7.5Hz, -CH ₂ CO-) 1.8 ~ 1.2 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₄₄ H ₅₄ O ₈ Calculated value C 74.38% H 7.66% Actual value C 74.31% H 7.62%

Compound (8) [α] ²⁵ _D + 21.6 ° (C 1.10, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 1732, 1497, 1454, 1100 ¹ H-NMR (CDCl ₃ , δppm): 7.5 ~ 7.1 (20H, m, ph), 4.73 (1H, s, H-1), 3.66 (3H, s, OCH ₃ ), 2.30 (2H, t, J = 7.5Hz, -CH ₂ CO-) 1.7 ~ 1.2 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₄₄ H ₅₄ O ₈ Calculated value C 74.38% H 7.66% Actual value C 73.95% H 7.65%

Example 1 6-Methoxycarbonylhexyl β-D-mannopyranoside (9) To a mixed solution of 350 mg (0.523 mmol) of compound (5) (15 ml of methanol and 5 ml of acetic acid) was added 10% of palladium carbon (100 mg), and the mixture was stirred at room temperature for 4. ² under pressure of 5 kg / cm 2
Hydrogenated for 4 hours. After removing the catalyst by filtration, the filtrate was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (chloroform: methanol = 10: 1) to obtain 130 mg of compound (9) as crystals.

Mp 113-114 ° C. [α] _D -44.4 ° (C 0.54, MeOH) IR (KBr) cm ⁻¹ : 1732, 1084, 1026, ¹ H-NMR (CD ₃ OD, δppm): 4.48 (1H , d, J = 0.5Hz, H-1) 3.64 (3H, s, OMe), 2.33 (2H, t, J = 7.5Hz, -CH ₂ CO-) 1.7 ~ 1.25 (8H, m,-(CH ₂ ) _4- )

Elemental analysis C ₁₄ H ₂₆ O ₈ 0.5 H ₂ O Calculated value C 50.74% H 8.21% Actual value C 50.60% H 7.94%

Example 2 6-Methoxycarbonylhexyl α-D-mannopyranoside (10) 130 mg (0.19 mmol) of compound (6) was treated in the same manner as in Example 1 in a methanol-acetic acid mixture to prepare 10% palladium-carbon. Hydrogenation was carried out in the presence of the compound to effect de-O-benzylation to obtain 90 mg of compound (10).

[Α] ²⁶ _D + 50.0 ° (C 0.49, MeOH) IR (film) cm ⁻¹ : 3446, 1728, 1438, 1060 ¹ H-NMR (CD ₃ OD, δppm): 4.73 (1H, d, J = 1.5Hz, H-1) 3.65 (3H, s, OMe), 2.33 (2H, t, J = 7.5Hz, -CH 2 CO-) 1.7 ~1.35 (8H, m, - (CH 2) 4 - )

Elemental analysis C ₁₄ H ₂₆ O ₈ 0.5 H ₂ O Calculated value C 50.74% H 8.21% Measured value C 50.99% H 8.01%

Example 3 8-Methoxycarbonyloctyl β-D-mannopyranoside (11) This compound was synthesized by the method described in Carbohydr. Res., 156, 1 (1986).

Example 4 8-Methoxycarbonyloctyl α-D-mannopyranoside (12) To 15 ml of a methanol solution of 300 mg (0.483 mmol) of the compound (8), 54 mg of 10% palladium carbon was added, and hydrogenated at room temperature for 8 hours. . After removing the catalyst by filtration, the filtrate was concentrated to dryness under reduced pressure. The obtained crystals are ethyl acetate
The crystals are collected by filtration, washed with an ether mixture, and the compound (12) is obtained.
120 mg (71% yield) was obtained.

Mp 76-77 ° C. [α] ²⁵ _D + 52.7 ° (C 0.423, MeOH) IR (KBr) cm ⁻¹ : 1735, 1728, ¹ H-NMR (CD ₃ OD, δppm): 4.73 (1H, d, J = 1.6Hz, H-1) 3.65 (3H, s, OMe), 2.32 (2H, t, J = 7.5Hz, -CH ₂ CO-) 1.7 ~ 1.2 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₁₆ H ₃₀ O ₈ Calculated value C 54.84% H 8.63% Actual value C 54.73% H 8.55%

Reference Example 4 8-Methoxycarbonyloctyl 3,4,6-tri-
O-acetyl-2-azido-2-deoxy-α-D-mannopyranoside (14) and 8-methoxycarbonyloctyl 3,4,6-tri-O-acetyl-2-azido-2-deoxy-β-D- Mannopyranoside (15)

8-Methoxycarbonyloctanol 24
After dissolving 1 mg (1.27 mmol) in 20 ml of dry toluene, 5 ml of the solvent was distilled off under a nitrogen stream. After cooling this solution in an ice bath, Carbohydr. Res., 136, 153 (1985)
505 mg (1.27 mmol) of 3,4,6-tri-O-acetyl-2-azido-2-deoxy-α-D-mannopyranosyl bromide (13) synthesized by the method described in 5 ml of dry toluene. The solution dissolved in was added. After adding 500 mg of the powdery molecular sieve 4A and stirring for 15 minutes, 500 mg of silver silicate was added and further stirred at the same temperature for 18 hours. The reaction mixture was filtered through Celite, and the filtrate was concentrated to dryness under reduced pressure to give a syrup-like residue.
The residue was hexane: ethyl acetate = using a rover column
Elution with 15 ml of each fraction was performed at 3: 1. Fraction No. 33-4
Compounds (14) 250 mg (yield 45.2%)
I got

Compound (14) (α) ²³ _D + 67.0 ° (C 1.3, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 2028, 1745, 1439, 1370, 1053 ¹ H-NMR (CDCl ₃ , δppm) : 5.42~ 5.25 (2H, m, H-3, H-4), 4.83 (1H, d, J 1,2 = 1.6Hz, H-1), 4.25 (1H, dd, J 5,6a = 4.8Hz , J _{6a, 6b} = 12Hz, H-6a), 4.09 (1H, dd, J _5,6b = 2.2Hz, H-6b), 4.01 (1H, dd, J _2.3 = 1.4Hz, H-2), 3.88 ~ 3.86 (1H, m, H-5), 3.67 (3H, s, OCH ₃ ), 2.30 (2H, t, J = 6Hz, -CH ₂ CO-), 2.10 (6H, s, 2 × OAc), 2.05 (3H, s, OAc), 1.2 ~ 1.7 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₂₂ H ₃₅ N ₃ O ₁₀ Calculated value C 52.68% H 7.03% N 8.38% Actual value C 52.68% H 6.90% N 8.47%

Fraction No. 59-84 was collected to obtain compound (1
5) 273 mg (yield 42.6%) was obtained.

Compound (15) (α) ²³ _D -75.5 ° (C 0.83, CHCl ₃ ) IR (CHCl ₃ ) cm ^-1 : 2028, 1745, 1439, 1370, 1053 ¹ H-NMR (CDCl ₃ , δppm) : 5.25 (1H, t, J _3.4 = 10Hz, H-4), 4.98 (1H, dd, J _2.3 = 4Hz, H-3), 4.66 (1H, d, J = 1.5Hz, H-1), 4.28 (1H, dd, J _5,6b = 4.8Hz, J _{6a, 6b} = 12Hz, H-6a), 4.13 (1H, dd, J _5.6b = 2.2Hz, H-6b), 2.30 (2H, t, J = 7Hz, -CH ₂ CO-), 2.11 (3H, s, OAc), 2.10 (3H, s, OAc), 2.04 (3H, s, OAc), 1.2 ~ 1.7 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₂₂ H ₃₅ N ₃ O ₁₀ Calculated value C 52.68% H 7.03% N 8.38% Actual value C 52.55% H 7.08% N 8.39%

Reference Example 5 6-Methoxycarbonylhexyl 3,4,6-tri-
O-acetyl-2-azido-2-deoxy-α-D-mannopyranoside (16)

271 mg (1.85 mmol) of 6-methoxycarbonylhexanol (2) and 3,4,6
-Tri-O-acetyl-2-azido-2-deoxy-α
-D-mannopyranosyl bromide (13) 730 mg
(1.85 mmol) was condensed with toluene in the same manner as in Reference Example 4 in toluene in the presence of molecular sieve 4A using silver silicate to give 424 mg of compound (16) (yield 4
9.9%).

[Α] ²⁵ _D + 67.8 ° (C 0.72, CHCl ₃ ) IR (film) cm ⁻¹ : 2110, 1747, 1436, 1369, 1228, 1053 ¹ H-NMR (CDCl ₃ , δppm): 5.44 ~ 5.26 (2H, m, H- 3, H-4), 4.82 (1H, d, J 1.2 = 1.5Hz, H-1), 4.24 (1H, dd, J 5,6a = 4.5Hz, J 6a, 6b = 10Hz, H-6a), 4.08 (1H, dd, J _5,6b = _2.5Hz , H-6b), 4.00 (1H, dd, J _2.3 = 3.5Hz, H-2), 3.84 ~ 3.87 (1H, m, H-5), 2.32 (2H, t, J = 7.5Hz, -CH ₂ CO-), 2.09 (6H, s, 2 × OAc), 2.04 (3H, s, OAc), 1.70 ~ 1.25 (8H _{, m, - (CH 2)} 4 -)

Elemental analysis C ₁₉ H ₂₉ N ₃ O ₁₀ Calculated value C 49.67% H 6.36% N 9.15% Actual value C 49.33% H 6.05% N 9.01%

Reference Example 6 8-Methoxycarbonyloctyl 2-azido-2-deoxy-α-D-mannopyranoside (17)

0.1 ml of a 1N sodium methoxide-methanol solution was added to 7 ml of a methanol solution of 283 mg (0.564 mmol) of the compound (14), and the mixture was allowed to stand at room temperature for 4 hours. Ion exchange resin Amberlite IR-1 for reaction liquid
After neutralizing by adding 20B (H ⁺ type), the resin was filtered off,
It was washed with a small amount of methanol. The filtrate and washings were combined and the solvent was distilled off under reduced pressure to obtain a syrupy residue. The residue was dissolved in methanol, activated carbon was added, and the mixture was left for a while. After filtering using a Millipore filter, the filtrate was concentrated to dryness under reduced pressure to give 210 mg of compound (17) (quantitative).
Was obtained as a syrupy material.

[Α] ²⁴ _D + 68.6 ° (C 0.614, MeOH) IR (CHCl ₃ ) cm ⁻¹ : 3600, 3440, 2028, 1730 ¹ H-NMR (CD ₃ OD, δppm): 4.77 (1H, d , J _1,2 = 1.5Hz, H-1), 3.65 (3H, s, OMe), 2.32 (2H, t, J = 7Hz, -CH ₂ CO-), 1.7 ~ 1.2 (12H, m,-( CH ₂ ) _6- )

Elemental analysis C ₁₆ H ₂₉ N ₃ O ₇ Calculated value C 51.19% H 7.79% N 11.19% Actual value C 50.87% H 7.66% N 11.02%

Reference Example 7 8-methoxycarbonyloctyl 2-azido-2-deoxy-β-D-mannopyranoside (18) Compound (15) 273 mg (0.54 mmol) was added in methanol in the same manner as in Reference Example 6. De-O-acetylation with a catalytic amount of sodium methoxide to give compound (18)
133 mg were obtained as crystals.

Mp 47-49 ° C. [α] ²⁴ _D -83.8 ° (C 0.798, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 3420, 2028, 1732 ¹ H-NMR (CD ₃ OD, δppm): 4.68 (1H, d, J _1,2 = 1.0Hz, H-1), 3.65 (3H, s, OCH ₃ ), 2.31 (2H, t, J = 7Hz, -CH ₂ CO-), 1.7 ~ 1.2 (12H , m,-(CH ₂ ) _6- )

Elemental analysis C ₁₆ H ₂₉ N ₃ O ₇ Calculated value C 51.19% H 7.79% N 11.19% Actual value C 51.00% H 7.82% N 10.55%

Reference Example 8 6-Methoxycarbonylhexyl 2-azido-2-deoxy-α-D-mannopyranoside (19) 420 mg (0.914 mmol) of the compound (16) was added to methanol in the same manner as in Reference Example 6. De-O-acetylation with a catalytic amount of sodium methoxide to give compound (1
9) 265 mg (yield 87%) was obtained as a syrup-like substance.

[Α] ^22.5 _D + 75.8 ° (C 0.89, MeOH) IR (film) cm ⁻¹ : 3400, 2120, 1735, 1270, 1060 ¹ H-NMR (CD ₃ OD, δppm): 4.77 (1H, s, H-1), 3.65 (3H, s, OCH 3), 2.33 (2H, t, J = 7.5Hz, -CH 2 CO-), 1.28~1.70 (8H, m, - (CH 2) 4 - )

Example 5 8-Methoxycarbonyloctyl 2-acetamido-
2-Deoxy-α-D-mannopyranoside (20) (i) Compound (17) 180 mg (0.477 mmol)
0.5 ml of acetic anhydride was added to 10 ml of methanol solution of
Hydrogenated in the presence of 30% 10% palladium on carbon for 1 hour at room temperature. After removing the catalyst by filtration, the filtrate was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (chloroform: methanol = 10: 1) to obtain 85 mg of compound (20) (yield 45.4%) as an amorphous powder.

[Α] ²⁵ _D + 37.1 ° (C 0.421, MeOH) IR (CHCl ₃ ) cm ⁻¹ : 1732, 1655, 1439, 1375, 1130, 1070, 1027 ¹ H-NMR (CD ₃ OD, δppm) : 4.66 (1H, d, J _1.2 = 1.6Hz, H-1), 4.28 (1H, dd, J _3.4 = 4.8Hz, H-2), 3.65 (3H, s, OMe), 2.32 (2H, t, J = 7.5Hz, -CH ₂ CO-), 2.00 (3H, s, NAc), 1.25 ~ 1.70 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₁₈ H ₃₃ NO ₈・ 0.2H ₂ O Calculated value C 54.72% H 8.52% N 3.55% Measured value C 54.60% H 8.41% N 3.72%

(Ii) 210 mg (0.55) of compound (17)
(9 mmol) in 5 ml of ethanol, 380 mg (1.6 mmol) of nickel chloride hexahydrate in 10 ml of ethanol.
0.1 ml of the solution dissolved in was added, and then 63 mg (1.677 mmol) of sodium borohydride in ethanol solution 5
ml was added with stirring. After stirring at room temperature for 30 minutes,
Acetic acid was added to the reaction solution to neutralize it. Then acetic anhydride 0.2
ml was added and left at room temperature for 1 hour. The reaction mixture was concentrated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform: methanol = 10: 1) to obtain 147 mg of the same compound (20) as above (yield 67.
4%).

Example 6 8-Methoxycarbonyloctyl 2-acetamido-
2-Deoxy-β-D-mannopyranoside (21) 149 mg (0.396 mmol) of compound (18) was used in Example 5
As in the case of (ii), the azide group was reduced to an amino group with sodium borohydride in the presence of a catalytic amount of nickel chloride hexahydrate in ethanol, and then N-acetylated with acetic anhydride to obtain a compound ( 21) 101 mg (yield 65.1%) was obtained as a syrup-like substance.

[Α] ²⁴ _D -49.4 ° (C 0.842, MeOH) IR (KBr) cm ^-1 : 1740, 1645, 1550, 1100 ¹ H-NMR (CD ₃ OD, δppm): 4.60 (1H, d, J _1.2 = 1.6Hz, H-1), 4.42 (1H, dd, J _2.3 = 4.2Hz, H-2), 3.64 (3H, s, OMe), 2.30 (2H, t, J = 7.5Hz, -CH ₂ CO-), 2.01 (3H, s, NAc), 1.7 ~ 1.2 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₁₈ H ₃₃ NO ₈・ 0.3H ₂ O Calculated value C 54.47% H 8.53% N 3.53% Measured value C 54.55% H 8.36% N 3.59%

Example 7 6-Methoxycarbonylhexyl 2-acetamido-
2-Deoxy-α-D-mannopyranoside (22) 265 mg (0.795 mmol) of compound (19) was used in Example 5
As in the case of (ii), the azide group was reduced to an amino group with sodium borohydride in the presence of a catalytic amount of nickel chloride hexahydrate in ethanol, and then N-acetylated with acetic anhydride to obtain a compound ( 22) 228 mg (82% yield) was obtained as a syrupy substance.

[Α] ^23.5 _D + 38.3 ° (C 0.52, MeOH) IR (film) cm ⁻¹ : 3328, 1730, 1650, 1552, 1438, 1377 ¹ H-NMR (CD ₃ OD, δppm): 4.65 ( 1H, d, J _1.2 = 1.2Hz, H-1), 4.27 (1H, dd, J _2.3 = 4.3Hz, H-2), 3.65 (3H, s, OMe), 2.33 (2H, t, J = 7.5 _{Hz, -CH 2 CO-), 1.99} (3H, s, NAc), 1.70~ 1.35 (8H, m, - (CH 2) 4 -)

Elemental analysis C ₁₅ H ₂₇ NO ₈ · H ₂ O Calculated value C 49.04% H 7.96% N 3.81% Actual value C 49.34% H 8.02% N 3.65%

Reference Example 9 8-Methoxycarbonyloctyl 2-O- (2,3,
4,6-Tetra-O-benzyl-β-D-mannopyranosyl) -3,4,6-tri-O-benzyl-α-D-mannopyranoside (24) and 8-methoxycarbonyloctyl 2-O- (2. 3,4,6-Tetra-O-benzyl-α-D-mannopyranosyl) -3,4,6-tri-O-benzyl-α-D-mannopyranoside (25)

8-methoxycarbonyloctyl 3,
4,6-Tri-O-benzyl-α-D-mannopyranoside (23) (Carbohydr. Res., 156, 1 (1986)) 600
mg (0.966 mmol) and 602 mg (1.08 mmol) 2,3,4,6-tetra-O-benzyl-α-D-mannopyranosyl chloride (3) in anhydrous benzene (20 ml). Then, 2 g of powdery molecular sieve 4A was added under a nitrogen stream, and the mixture was stirred at room temperature for 30 minutes. Next, 1.0 g of silver silicate was added, and the mixture was stirred at room temperature for 48 hours. The reaction mixture was filtered through Celite, and the filtrate was concentrated under reduced pressure to dryness to obtain 1.09 g of a residue. The residue was separated and purified using a Rover column to give compound (24) 484.
mg (yield 42.3%) and compound (25) 120 mg
(Yield 10%) was obtained as a syrup-like substance.

Compound (25) [α] ²⁵ _D + 15.0 ° (C 0.808, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 1732, 1498, 1455, 1315, 1105, 1029 ¹ H-NMR (CDCl ₃ , δppm): 7.4 ~ 7.1 (35H, m, ph), 5.18 (1H, d, J = 1.0Hz, H-1 or H-1 '), 4.86 (1H, d, J = 1.0Hz, H-1 or H-1 ′), 3.66 (3H, s, OMe), 2.30 (2H, t, J = 7.5Hz, -CH ₂ CO-), 1.7 to 1.15 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₇₁ H ₈₂ O ₁₃ Calculated value C 74.58% H 7.23% Actual value C 74.46% H 7.22%

Compound (24) [α] ²⁵ _D -38.5 ° (C 0.924, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 1731, 1498, 1452, 1362, 1100, 1065 ¹ H-NMR (CDCl ₃ , δppm): 7.6 ~ 7.0 (35H, m, ph), 4.90 (1H, s, H-1 or H-1 ′), 3.67 (3H, s, OMe), 2.30 (2H, t, J = 7.5Hz, -CH ₂ CO-), 1.7 ~ 1.2 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₇₁ H ₈₂ O ₁₃ Calculated value C 74.58% H 7.23% Actual value C 74.32% H 7.30%

Example 8 8-Methoxycarbonyloctyl 2-O- (β-D-
Mannopyranosyl) -α-D-mannopyranoside (2
6) 480 mg (0.42 mmol) of compound (24) was hydrogenated in the presence of 10% palladium carbon in methanol to de-O-benzylate to give 181 mg of compound (26) (yield 84
%) As a colorless powder.

[Α] ²⁴ _D -4.9 ° (C 0.658, MeOH) IR (KBr) cm ⁻¹ : 3400, 1740, 1060 ¹ H-NMR (CD ₃ OD, δppm): 4.86 (1H, overlapped with CD ₃ OH, H-1 or H-1 ′), 4.65 (1H, d, J = 0.8Hz, H-1 ′ or H-1) 3.65 (3H, s, OMe), 2.31 (2H, t, J = 7.5 Hz, -CH ₂ CO-), 1.7 ~ 1.2 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₂₂ H ₄₀ O ₁₃ · H ₂ O Calculated value C 49.80% H 7.98% Actual value C 50.10% H 7.67%

Example 9 8-Methoxycarbonyloctyl 2-O- (α-D-
Mannopyranosyl) -α-D-mannopyranoside (2
7) This compound was synthesized by the method described in Carbohydr. Res., 156, 1 (1986).

Reference Example 10 8-Methoxycarbonyloctyl 2,3,4-tri-
O-benzyl-β-D-mannopyranoside (28)

(I) J. Carbohydr. Chem., 6, 645-660
(1987) 1,6-di-O-acetyl-2,3,4-tri-O-benzyl-α-D-mannopyranoside 5.00 g (9.35 mmol) of methylene chloride in 60 ml. Hydrogen chloride gas was blown into the solution at −20 ° C. to be saturated. After the reaction solution was returned to room temperature and left for 2 hours, it was concentrated under reduced pressure and 6-O-acetyl-2,3,4-tri-O-benzyl-α-D-mannopyranosyl chloride 4.69 g ( Yield 98.2%) was obtained as a syrup-like substance. To a solution of 4.69 g of this chloride in 100 ml of 1,2-dichloroethane, 7.09 g of mercuric cyanide (2
8.1 mmol), 30 g of powdered molecular sieve 4A
And 8-methoxycarbonyloctanol 1.76g
(9.35 mmol) was added, and the mixture was heated at 80 ° C. in an argon stream.
And stirred for 22 hours. After removing the precipitate by filtration, the filtrate was concentrated to dryness under reduced pressure to obtain a syrupy residue. The residue was subjected to silica gel column chromatography (n-hexane: ethyl acetate =
7: 1) and purified with 8-methoxycarbonyloctyl
6-O-acetyl-2,3,4-tri-O-benzyl-
3.24 g (yield 52.2%) of an α / β-D-mannopyranoside mixture was obtained.

(Ii) 2.45 g (3.
(70 mmol) in 50 ml of methanol was added with 5 ml of 2N sodium methylate and stirred at room temperature for 1.5 hours.
The reaction solution was neutralized with Amberlite IR-120B (H ⁺ type), and the resin was filtered off. The filtrate was concentrated to dryness under reduced pressure to obtain a syrupy residue. The residue was purified by silica gel column chromatography (toluene: ethyl acetate = 16: 1) to obtain 1.20 g of compound (28) (yield 52.2%). At the same time, 834 mg (yield 36.2%) of the known compound (29) was obtained from Carbohydr. Res., 161, 195 (1987).

[Α] ²⁴ _D -38.7 ° (C 1, CHCl ₃ ) IR (KBr) cm ⁻¹ : 3500, 2930, 2850, 1740 ¹ H-NMR (CDCl ₃ , δppm): 2.28 (2H, t, J = 7.6Hz, CH ₂ COOCH ₃ ), 3.65 (3H, s, OCH ₃ )

Reference Example 11 8-Methoxycarbonyloctyl 2,3,4-tri-
O-benzyl-6-O- (2,3,4,6-tetra-O
-Benzyl-β-D-mannopyranosyl) -α-D-mannopyranoside (30) and 8-methoxycarbonyloctyl 2,3,4-tri-O-benzyl-6-O-
(2,3,4,6-Tetra-O-benzyl-α-D-mannopyranosyl) -α-D-mannopyranoside (31)

(I) 2,3,4,6-tetra-O-benzyl-α-D-mannopyranosyl chloride (3) 26
5 mg (0.47 mmol) and compound (29) 300 mg
5 ml of (0.483 mmol) of 1,2-dichloroethane
496 mg (1.93 mmol) of silver triflate and 337 mg (2.90) of 1,1,3,3-tetramethylurea were added to the solution.
Was added and the mixture was stirred at room temperature for 3 hours in an argon stream. Further, 265 mg (0.474 mmol) of compound (3) was added and stirred for 2 days. The syrup obtained by concentrating the reaction solution under reduced pressure was purified by silica gel column chromatography (toluene: ethyl acetate = 30: 1) to obtain 151 mg of compound (30) (yield 27.3%) and 273 mg of compound (31) ( The yield was 42.9%).

Compound (30) [α] ²⁴ _D -5.3 ° (C 1, CHCl ₃ ) IR (KBr) cm ⁻¹ : 2930, 2850, 1740 ¹ H-NMR (CDCl ₃ , δppm): 2.28 (2H, t, J = 7.6Hz, CH ₂ COOCH ₃ ), 3.65 (3H, s, OCH ₃ )

Compound (31) [α] ²⁴ _D + 56.2 ° (C 1, CHCl ₃ ) IR (KBr) cm ⁻¹ : 2930, 2850, 1740 ¹ H-NMR (CDCl ₃ , δppm): 2.28 (2H, t, J = 7.6Hz, CH ₂ COOCH ₃ ), 3.65 (3H, s, OCH ₃ ), 4.70 (1H, d, J = 1.8Hz, H-1)

(Ii) 90 mg (0.161 mmol) of compound (3) and 100 mg (0.161 mmol) of compound (29) in 2.5 ml of 1,2-dichloroethane were added to 325 mg of silver silicate and 1.5 g of powdered molecular sieve 4A. Was added and stirred for 3 hours. Compound (3) 90mg
(0.161 mmol) was added and stirred for 21 hours. After removing the precipitate by filtration, the filtrate was concentrated to dryness under reduced pressure to obtain a syrupy residue. The residue was purified by silica gel column chromatography (toluene: ethyl acetate = 30: 1) to obtain 103 mg of the same compound (30) (yield 55.9%) and 44 mg of compound (31) (yield 23.9%). ) Got.

Reference Example 12 8-Methoxycarbonyloctyl 2,3,4-tri-
O-benzyl-6-O- (2,3,4,6-tetra-O
-Benzyl-β-D-mannopyranosyl) -β-D-mannopyranoside (32) and 8-methoxycarbonyloctyl 2,3,4-tri-O-benzyl-6-O-
(2,3,4,6-Tetra-O-benzyl-α-D-mannopyranosyl) -β-D-mannopyranoside (33)

(I) Compound (3) 180 mg (0.322)
Mmol) and 200 mg (0.322 mmol) of compound (28) in the same manner as in Reference Example 11 (i),
1,1,3,3- in 1,2-dichloroethane 5 ml solution
Condensation using silver triflate in the presence of tetramethylurea gave compound (32) 25 mg (yield 6.8%) and compound (33) 228 mg (yield 61.9%).

Compound (32) [α] ²⁴ _D -41.9 ° (C 0.5, CHCl ₃ ) IR (KBr) cm ⁻¹ : 2930, 2850, 1740 ¹ H-NMR (CDCl ₃ , δppm): 2.28 (2H, t, J = 7.6Hz, CH ₂ COOCH ₃ ), 3.65 (3H, s, OCH ₃ )

Compound (33) [α] ²⁴ _D -5.2 ° (C 1, CHCl ₃ ) IR (KBr) cm ⁻¹ : 2930, 2850, 1740 ¹ H-NMR (CDCl ₃ , δppm): 2.28 (2H, t, J = 7.6Hz, CH ₂ COOCH ₃ ), 3.65 (3H, s, OCH ₃ )

(Ii) Compound (3) 180 mg (0.322)
1) and 200 mg (0.322 mmol) of the compound (28) in the same manner as in the method of Reference Example 11 (ii).
196 mg (53.2% yield) of the same compound (32) and 94 mg (yield 25.25%) of the same compound as above, condensed by using silver silicate in the presence of powdered molecular sieve 4A in a 5 ml solution of dichloroethane. 5%) was obtained.

Example 10 8-Methoxycarbonyloctyl 6-O- (β-D-
Mannopyranosyl) -α-D-mannopyranoside (3
4) To a solution of 117 mg (0.102 mmol) of compound (30) in 20 ml of methanol was added 100 mg of 10% palladium carbon, and catalytic reduction was carried out for 2 days using a Parr apparatus (50 psi).
After removing the catalyst by filtration, the filtrate was concentrated to dryness under reduced pressure to obtain a syrupy residue. The residue was subjected to silica gel column chromatography (chloroform: methanol: concentrated aqueous ammonia = 5:
5: 1) and compound (34) 45.1 mg (yield 8
6.3%) was obtained.

[Α] ²⁴ _D + 0.1 ° (C 0.9, MeOH) ¹ H-NMR (CD ₃ OD, δppm): 2.31 (2H, t, J = 7.6Hz, CH ₂ COOCH ₃ ), 3.65 (3H, s, OCH ₃ ), 4.59 (1H, s, H- 1 ′) 4.70 (1H, d, J = 1.8Hz, H-1) MS (SIMS): m / z 535 (M + Na) ⁺

Example 11 8-Methoxycarbonyloctyl 6-O- (α-D-
Mannopyranosyl) -α-D-mannopyranoside (3
5) Example 1 containing 183 mg (0.160 mmol) of compound (31)
In the same manner as in the case of 0, hydrogenation was performed in the presence of 10% palladium carbon in methanol, and de-O-benzylation was performed to obtain 38.6 mg of compound (35) (yield 47.1%).

[Α] ²⁴ _D + 55.1 ° (C 0.8, MeOH) ¹ H-NMR (CD ₃ OD, δppm): 2.31 (2H, t, J = 7.6Hz, CH ₂ COOCH ₃ ), 3.65 (3H, s, OCH ₃ ), 4.70 (1H, d, J = 1.8Hz, H-1) MS (SIMS): m / z 535 (M + Na) ⁺

Example 12 8-Methoxycarbonyloctyl 6-O- (β-D-
Mannopyranosyl) -β-D-mannopyranoside (3
6) 151 mg (0.132 mmol) of compound (32) was used in Example 1
In the same manner as in the case of 0, hydrogenation was performed in the presence of 10% palladium carbon in methanol, and de-O-benzylation was performed to obtain 49.0 mg of compound (36) (yield 72.4%).

[Α] ²⁴ _D -53.3 ° (C 1, MeOH) ¹ H-NMR (CD ₃ OD, δppm): 2.31 (2H, t, J = 7.6Hz, CH ₂ COOCH ₃ ), 3.65 (3H, s, OCH ₃ ), 4.48 (1H, s, H-1) 4.61 (1H, s, H-1 ′) MS (SIMS): m / z 535 (M + Na) ⁺

Example 13 8-Methoxycarbonyloctyl 6-O- (α-D-
Mannopyranosyl) -β-D-mannopyranoside (3
7) 196 mg (0.171 mmol) of compound (33) was used in Example 1
In the same manner as in the case of 0, hydrogenation was performed in the presence of 10% palladium carbon in methanol, and de-O-benzylation was performed to obtain 49.0 mg of compound (37) (yield 72.4%).

[Α] ²⁴ _D -53.3 ° (C 1, MeOH) ¹ H-NMR (CD ₃ OD, δppm): 2.31 (2H, t, J = 7.6Hz, CH ₂ COOCH ₃ ), 3.65 (3H, s, OCH ₃ ), 4.48 (1H, s, H-1) MS (SIMS): m / z 535 (M + Na ⁺ )

Reference Example 13 8-Methoxycarbonyloctyl 2-O- (3,4,
6-tri-O-acetyl-2-azido-2-deoxy-
α-D-mannopyranosyl) -3,4,6-tri-O-
Benzyl-α-D-mannopyranoside (38) 8-Methoxycarbonyloctyl 3,4,6-tri-O-benzyl-α-D-mannopyranoside synthesized by the method described in Carbohydr. Res., 156, 1 (1986). (23) 7
20 ml of a 93 mg (1.278 mmol) toluene solution was ice-cooled under a nitrogen stream. To this, add 800 mg of powdered molecular sieve 4A and 800 mg of silver silicate, and add 30
Stir for minutes. Then 3,4,6-tri-O-acetyl-2-azido-2-deoxy-α-D-mannopyranosyl bromide (13) 505 mg (1.278 mmol)
5 ml of a toluene solution of was added, and the mixture was stirred at the same temperature for 48 hours. The reaction mixture was filtered through Celite, and the filtrate was concentrated under reduced pressure to dryness to obtain 1.22 g of a residue. The residue was purified using a Rover column (hexane: ethyl acetate = 3: 1) to give 557 mg of compound (38) (yield 59.7%).
I got

[Α] ²⁵ _D + 37.4 ° (C 0.901, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 2028, 1745, 1452, 1369, 1045. ¹ H-NMR (CDCl ₃ , δppm): 7.5 ~ 7.1 (15H, m, ph), 5.43 (1H, dd, J _{3 ', 4'} = 9.4Hz, J _{2 ', 3'} = 3.7Hz, H- 3 '), 5.29 (1H, t, H-4 ), 4.98 (1H, d, J _{1 ', 2'} = 1.8Hz, H-1 '), 4.85 (1H, d, J _1,2 = 1.7Hz, H-1), 3.66 (3H, s, OMe ), 2.30 (2H, t, J = 7Hz, -CH ₂ CO-), 2.10 (3H, s, OAc), 2.09 (3H, s, OAc), 1.99 (3H, s, OAc), 1.8 ~ 1.2 ( 12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₄₉ H ₆₃ N ₃ O ₁₅ Calculated value C 62.94% H 6.79% N 4.49% Actual value C 62.76% H 6.83% N 4.61%

Reference Example 14 8-Methoxycarbonyloctyl 2-O- (2-azido-2-deoxy-α-D-mannopyranosyl) -3,
4,6-Tri-O-benzyl-α-D-mannopyranoside (39)

To 20 ml of a methanol solution of 557 mg (0.596 mmol) of the compound (38) was added 0.15 ml of a 1M sodium methoxide-methanol solution, and the mixture was allowed to stand at room temperature for 2 hours. Ion exchange resin Amberlite IR for reaction liquid
After adding -120B (H ⁺ type) for neutralization, the resin was filtered off and washed with a small amount of methanol. The filtrate and washings were combined and concentrated to dryness under reduced pressure to obtain 447 mg (yield 93%) of compound (39) as a syrup-like substance.

[Α] ²⁴ _D + 46.7 ° (C 0.559, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 3600, 3460, 2027, 1731, 1498, 1453, 1065 ¹ H-NMR (CD ₃ OD, δppm ): 7.40 ~ 7.15 (15H, ph), 4.99 (1H, d, J _{1 ', 2'} = 1.4Hz, H-1 '), 4.93 (1H, d, J _1,2 = 1.8Hz, H-1 ), 3.62 (3H, s, OMe), 2.30 (2H, t, J = 7Hz, -C H ₂ C O-), 1.7 ~ 1.2 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₄₃ H ₅₇ N ₃ O ₁₂ Calculated value C 63.92% H 7.11% N 5.20% Actual value C 63.17% H 7.09% N 5.22%

Reference Example 15 8-Methoxycarbonyloctyl 2-O- (2-acetamido-2-deoxy-α-D-mannopyranosyl)
-3,4,6-Tri-O-benzyl-α-D-mannopyranoside (40)

0.08 ml of an ethanol solution of 0.16M nickel chloride hexahydrate was added to 8 ml of an ethanol solution of 310 mg (0.384 mmol) of the compound (39), and then 44 mg of sodium borohydride was stirred at room temperature. (1.15
A solution prepared by dissolving (mmol) in 5 ml of ethanol was added. The reaction mixture was stirred for 40 minutes and then acetic acid was added to neutralize it. Next, 0.2 ml of acetic anhydride was added and the mixture was left at room temperature for 1 hour. The reaction mixture was concentrated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform: methanol = 10: 1) to give compound (40) 184 mg.
(Yield 58.2%) was obtained as a syrup-like substance.

[Α] ²⁴ _D + 42.3 ° (C 0.522, MeOH) ¹ H-NMR (CD ₃ OD, δppm): 7.40 to 7.10 (15H, m, ph), 4.95 (1H, d, J = 1.8Hz , H-1 or H- 1 '), 4.93 (1H, d, J = 1.6Hz, H-1' or H-1), 3.62 (3H, s, OMe), 2.28 (2H, t, J = 7Hz , -C H ₂ C O-), 2.01 (3H, s, NAc), 1.7 ~ 1.2 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₄₅ H ₆₁ NO ₁₃ Calculated value C 65.59% H 7.46% N 1.70% Actual value C 65.23% H 7.33% N 1.95%

Example 14 8-Methoxycarbonyloctyl 2-O- (2-acetamido-2-deoxy-α-D-mannopyranosyl)
-Α-D-mannopyranoside (41)

To 15 ml of a solution of 170 mg (0.206 mmol) of compound (40) in methanol was added 10% palladium-
50 mg of carbon was added and hydrogenated at room temperature for 27 hours. After filtering the reaction mixture, the filtrate was concentrated to dryness under reduced pressure to obtain 86 mg (yield 75.4%) of compound (41) as an amorphous powder.

[Α] ²⁵ _D + 49.3 ° (C 0.708, MeOH) IR (KBr) cm ⁻¹ : 3400, 1740, 1650, 1545, 1125, 1060, 1020 ¹ H-NMR (CD ₃ OD, δppm): 5.01 (1H, s, H-1), 4.90 (1H, d, J _{1 ', 2'} = 1.5Hz, H-1 '), 4.44 (1H, dd, J _{2', 3 '} = 4.6Hz, H -2 ′), 3.65 (3H, s, OMe), 2.31 (2H, t, J = 7Hz, -C H ₂ C O-), 2.00 (3H, s, NAc), 1.7 ~ 1.2 (12H, m, -(CH ₂ ) _6- )

Elemental analysis C ₂₄ H ₄₃ NO ₁₃・ H ₂ O Calculated value C 50.43% H 7.94% N 2.45% Actual value C 50.52% H 7.76% N 2.62%

Reference Example 16 8-Methoxycarbonyloctyl 3,6-di-O-acetyl-2,4-di-O-benzyl-α-D-mannopyranoside (43)

3,6-di-O-acetyl-2,4-di-synthesized by the method described in Tetrahedron, 37, 2779 (1981).
After dissolving 3.24 g (7.58 mmol) of O-benzyl-α-D-mannopyranosyl chloride (42) and 1.43 g (7.58 mmol) of 8-methoxycarbonyloctanol in 60 ml of anhydrous benzene, Under a nitrogen stream, 10 ml of solvent was distilled off. After cooling to room temperature, 2.0 g of powdery molecular sieve 4A was added and stirred for 15 minutes. Then add 3.25 g of silver silicate and add 1 at room temperature.
Stir for 8 hours. The reaction mixture was filtered through Celite, and the filtrate was concentrated under reduced pressure to dryness. The residue (4.51 g) was purified using a Rover column (hexane: ethyl acetate = 3: 1) to obtain 2.93 g (yield 63%) of compound (43) as a syrup-like substance.

[Α] ²⁵ _D +4.4 ° (C 1.128, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 1735, 1455, 1365, 1075. ¹ H-NMR (CDCl ₃ , δppm): 7.45 to 7.2 ( 10H, m, ph), 5.25 (1H, dd, J _2,3 = 3.4Hz, J _3,4 = 8.8Hz, H-3), 4.82 (1H, d, J = 1.6Hz, H-1), 3.67 (3H, s, OMe). 2.31 (2H, t, J = 7.5Hz, -C H ₂ C O-), 2.08 (3H, s, OAc), 2.00 (3H, s, OAc), 1.7 ~ 1.2 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₃₄ H ₄₆ O ₁₀ Calculated value C 66.43% H 7.54% Actual value C 66.10% H 7.41%

Reference Example 17 8-Methoxycarbonyloctyl 2,4-di-O-benzyl-α-D-mannopyranoside (44) 1M sodium methoxy was added to a solution of 1.48 g (2.40 mmol) of the compound (43) in 30 ml of methanol. 0.6 ml of de-methanol solution was added, and the mixture was left at room temperature for 8 hours. Ion exchange resin Amberlite IR-120B for reaction liquid
(H ⁺ type) was added for neutralization, and the resin was filtered off. The filtrate was concentrated to dryness under reduced pressure to obtain 1.34 g of residue. The residue was purified by silica gel column chromatography (hexane: ethyl acetate = 2: 1) to give compound (44) 876 mg.
(Yield 69%) was obtained as a syrup-like substance.

[Α] ²⁵ _D + 18.2 ° (C 0.626, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 3560, 1731, 1498, 1455, 1438, 1100, 1075 ¹ H-NMR (CDCl ₃ , δppm) : 7.45 ~ 7.20 (10H, m, ph), 4.84 (1H, d, J = 1.2Hz, H-1), 3.67 (3H, s, OMe), 2.30 (2H, t, J = 7.5Hz, -CH ₂ CO-), 1.7 ~ 1.2 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₃₀ H ₄₂ O ₈ Calculated value C 67.90% H 7.98% Actual value C 67.39% H 8.03%

Reference Example 18 8-Methoxycarbonyloctyl 6-O- (3,4,
6-tri-O-acetyl-2-azido-2-deoxy-
α-D-mannopyranosyl) -2,4-di-O-benzyl-α-D-mannopyranoside (45)

710 mg (1.339 mmol) of compound (44) and 757 mg (2.94 mmol) of silver triflate.
A solution of 1.4 ml (11.78 mmol) of tetra-N-methylurea in dichloromethane (15 ml) was cooled to -78 ° C under a stream of nitrogen, and then 3,4,6-tri-O-acetyl-2-. 5 ml of a dichloromethane solution of 1.01 g (2.678 mmol) of azido-2-deoxy-α-D-mannopyranosyl bromide (13) was added. Then, the cooling bath was removed, the reaction temperature was gradually raised to room temperature, the mixture was stirred for 18 hours, and then the reaction mixture was filtered through Celite. The filtrate was washed successively with aqueous sodium hydrogen carbonate solution and water, dried over magnesium sulfate, and the solvent was evaporated under reduced pressure to give a syrup-like residue. The residue was purified using a Rover column (hexane: ethyl acetate = 1: 1) to obtain 486 mg of compound (45) (yield 43%) as a syrup-like substance.

[Α] ²⁵ _D + 62.3 ° (C 0.925, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 3560, 2110, 1745, 1455, 1370, 1060 ¹ H-NMR (CDCl ₃ , δppm): 7.5 ~ 7.25 (10H, m, ph), 5.42 ~ 5.22 (2H, m, H-3 ', H-4'), 5.04 (1H, d, J = 1.6Hz, H-1 or H-1 '), 4.81 (1H, d, J = 1.4Hz, H-1 ′ or H-1), 3.66 (3H, s, OMe), 2.30 (2H, t, J = 7.5Hz, -CH ₂ CO-), 2.08 ( 3H, s, OAc), 2.06 (3H, s, OAc), 2.03 (3H, s, OAc), 1.7 ~ 1.2 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₄₂ H ₅₇ N ₃ O ₁₅・ 0.5H ₂ O Calculated value C 59.14% H 6.85% N 4.93% Measured value C 59.07% H 6.77% N 5.07%

Reference Example 19 8-Methoxycarbonyloctyl 6-O- (2-azido 2-deoxy-α-D-mannopyranosyl) -2,
4-di-O-benzyl-α-D-mannopyranoside (4
6)

470 mg (0.557 mmol) of the compound (45) was de-O-acetylated with a catalytic amount of sodium methoxide in methanol in the same manner as in Reference Example 14 to give 333 mg of the compound (46) (yield 83 .2%) as a syrup.

[Α] ²⁵ _D + 69.1 ° (C 0.739, MeOH) IR (CHCl ₃ ) cm ⁻¹ : 3560, 2120, 1731, 1075 ¹ H-NMR (CD ₃ OD, δppm): 7.5 to 7.2 (10H , m, ph), 3.64 (3H, s, OMe), 2.30 (2H, t, J = 7.5Hz, -CH ₂ CO-), 1.7 ~ 1.25 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₃₆ H ₅₁ N ₃ O ₁₂・ 0.6H ₂ O Calculated value C 59.34% H 7.22% N 5.77% Measured value C 59.04% H 7.02% N 5.80%

Reference Example 20 8-Methoxycarbonyloctyl 6-O- (2-acetamido-2-deoxy-α-D-mannopyranosyl)
-2,4-Di-O-benzyl-α-D-mannopyranoside (47)

320 mg (0.446 mmol) of the compound (46) was treated with sodium borohydride in the same manner as in Reference Example 15 in the presence of a catalytic amount of nickel chloride hexahydrate to convert an azide group into an amino group. And then N-acetylated with acetic anhydride to obtain 234 mg of compound (47) (yield 7
1.5%) was obtained as an amorphous powder.

[Α] ²⁵ _D + 54.7 ° (C 0.569, MeOH) IR (CHCl ₃ ) cm ⁻¹ : 1730, 1660, 1455, 1075. ¹ H-NMR (CD ₃ OD, δppm): 7.5 to 7.2 ( 10H, m, ph), 4.41 (1H, dd, J _{1 ', 2'} = 1.2Hz, J _{2 ', 3'} = 5Hz, H-2 '), 3.64 (3H, s, OMe), 2.31 (2H , t, J = 7.5Hz, -CH ₂ CO-), 2.00 (3H, s, NAc), 1.7 ~ 1.25 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₃₈ H ₅₅ NO ₁₃・ H ₂ O Calculated value C 60.70% H 7.64% N 1.86% Actual value C 60.84% H 7.56% N 2.02%

Example 15 8-Methoxycarbonyloctyl 6-O- (2-acetamido-2-deoxy-α-D-mannopyranosyl)
-Α-D-mannopyranoside (48) Compound (47) 230 mg (0.313 mmol) was used in Example 1
In the same manner as in the case of 4, hydrogenation was carried out in the presence of 10% palladium-carbon in methanol to carry out de-O-benzylation to obtain 118 mg of compound (48) as an amorphous powder.

[Α] ²⁴ _D + 56.6 ° (C 0.758, MeOH) IR (KBr) cm ⁻¹ : 1740, 1650, 1545. ¹ H-NMR (CDCl ₃ , δppm): 4.74 (1H, d, J _{1 ', 2'} = 1.2Hz, H- 1 '), 4.71 (1H, d, J _1,2 = 1.4Hz, H-1), 4.36 (1H, dd, J _{2', 3 '} = 5Hz, H- 2 ′), 3.65 (3H, s, OMe), 2.32 (2H, t, J = 7.5Hz, -C H ₂ C O-), 2.01 (3H, s, NAc), 1.70 ~ 1.25 (12H, m, -(CH ₂ ) _6- )

Elemental analysis C ₂₄ H ₄₃ NO ₁₃・ H ₂ O Calculated value C 50.43% H 7.94% N 2.45% Actual value C 50.55% H 7.76% N 2.58%

Reference Example 21 8-Methoxycarbonyloctyl 2-O- (3,4,
6-tri-O-acetyl-2-deoxy-2-phthalimido-β-D-glucopyranosyl) -3,4,6-tri-O-benzyl-α-D-mannopyranoside (50)

Synthetic Methods for Carbohydrate Che
2,3,4-tri-O-acetyl-2-deoxy-2-phthalimido-β synthesized by the method described in mistry, 1976.
-D-glucopyranosyl bromide (49) 410 mg
(0.66 mmol) and compound (23) 427 mg
(0.85 mmol) was dissolved in 20 ml of anhydrous dichloromethane and then under a nitrogen stream, molecular sieve 4A (10 g)
Was added and stirred for 1 hour. Next, 400 mg of silver silicate was added, and the mixture was stirred at room temperature for 16 hours. The reaction mixture was filtered through Celite, and the filtrate was concentrated under reduced pressure to dryness to obtain 710 mg of a syrupy residue. The residue was purified by silica gel column chromatography (hexane: ethyl acetate = 2: 1) to give compound (50) 400 mg (yield 58.3%).

[Α] ²⁴ _D + 13.7 ° (C 0.35, CHCl ₃ ) ¹ H-NMR (CDCl ₃ , δppm): 6.9 to 7.9 (19H, m, aromatic H), 5.82 (1H, t, J = 10Hz , H-4 ′), 5.51 (1H, d, J = 8.5Hz, H-1 ′), 5.20 (1H, t, J = 10Hz, H-3 ′), 3.65 (3H, s, OMe), 2.28 (2H, t, J = 7.5Hz), 2.04 (6H, s, 2 × Ac), 1.87 (3H, s, Ac), 1.7 ~ 1.0 (12H, m,-(CH ₂ ) _6- )

Reference Example 22 8-Methoxycarbonyloctyl 2-O- (2-deoxy-2-phthalimido-β-D-glucopyranosyl)
-3,4,6-Tri-O-benzyl-α-D-mannopyranoside (51)

275 mg (0.265 mmol) of the compound (50) was de-O-acetylated with a catalytic amount of sodium methoxide in methanol in the same manner as in Reference Example 6 to obtain 240 mg of the compound (51).

¹ H-NMR (CDCl ₃ , δppm): 7.8 to 7.1 (19H, m, aromatic H), 5.30 (1H, d, J = 7.5Hz, H-1 ′), 4.49 (1H, d, J = 1.6Hz, H-1), 3.65 (3H, s, OMe), 2.28 (2H, t, J = 7.5Hz, -C H ₂ C O-), 1.7 ~ 1.1 (12H, m,-(CH ₂ ) _6- )

Reference Example 23 8-Methoxycarbonyloctyl 2-O- (2-acetamido-2-deoxy-β-D-glucopyranosyl)
-3,4,6-Tri-O-benzyl-α-D-mannopyranoside (52)

To a solution of the compound (51) (230 mg, 0.252 mmol) in anhydrous methanol (50 ml) was added hydrazine acetate (1.53 g, 16.6 mmol), and the mixture was heated under reflux for 5 hours under a nitrogen stream. After cooling to room temperature, the reaction solution was concentrated to dryness under reduced pressure. The residue was partitioned between an ethyl acetate layer and an aqueous layer. The organic layer was separated, washed with water, dried over magnesium sulfate, and the solvent was distilled off to obtain 210 mg of a residue.
The residue was dissolved in 20 ml of anhydrous methanol, and then acetic anhydride 2
ml was added and left at room temperature for 16 hours. The reaction mixture was concentrated to dryness under reduced pressure to give 236 mg of residue. The residue was purified by silica gel column chromatography (chloroform: methanol = 10: 1) to obtain 154 mg of compound (52).
(74.4%) was obtained.

[Α] ²⁵ _D -6.1 ° (C 0.676, MeOH) ¹ H-NMR (CD ₃ OD, δppm): 7.5-7.1 (15H, m, aromatic H), 4.53 (1H, d, J = 1.6 Hz, H-1), 3.62 (3H, s, OMe), 2.27 (2H, t, J = 7.5Hz, -C H ₂ C O-), 1.94 (3H, s, NAc), 1.7 ~ 1.2 (12H , m,-(CH ₂ ) _6- )

Example 16 8-Methoxycarbonyloctyl 2-O- (2-acetamido-2-deoxy-β-D-glucopyranosyl)
-Α-D-Mannopyranoside (53) 144 mg (0.174 mmol) of compound (52) was hydrogenated in the presence of 10% palladium carbon in methanol to effect de-O-benzylation to obtain 84 mg of compound (53).

[Α] ²⁴ _D +4.7 ° (C 0.838, MeOH) IR (KBr) cm ⁻¹ : 1740, 1645, 1560, 1065. ¹ H-NMR (CD ₃ OD, δppm): 4.77 (1H, d , J = 1.5Hz, H-1), 4.45 (1H, d, J = 8.1Hz, H-1 ′), 3.65 (3H, s, OMe), 2.32 (2H, t, J = 7.5Hz, -C H ₂ C O-), 1.99 (3H, s, NAc), 1.7 ~ 1.2 (12H, m,-(CH ₂ ) _6- )

Reference Example 24 8-Methoxycarbonyloctyl 6-O- (3,4,
6-tri-O-acetyl-2-deoxy-2-phthalimido-β-D-glucopyranosyl) -2,4-di-O-
Benzyl-α-D-mannopyranoside (54) and 8
-Methoxycarbonyloctyl 3,6-di-O-
(3,4,6-tri-O-acetyl-2-deoxy-2
-Phthalimido-β-D-glucopyranosyl) -2,4
-Di-O-benzyl-α-D-mannopyranoside (5
5)

To 25 ml of a solution of 564 mg (1.06 mmol) of compound (44) in anhydrous benzene was added 1.0 part of silver silicate.
g and powdery molecular sieve 4A (1.0 g) were added, and the mixture was stirred under a nitrogen stream at room temperature for 30 minutes. Then, 5 ml of an anhydrous benzene solution containing 1.05 g (2.12 mmol) of the compound (49) was added, and the mixture was stirred at room temperature for 16 hours. The reaction mixture was filtered through Celite, and the filtrate was concentrated under reduced pressure to dryness to give 1.6 g of a residue. The residue was purified using a Rover column (hexane: acetic acid S = 1: 1) to obtain 184 mg of compound (54) (yield 18.9%) and 693 mg of compound (55) (yield 48.9%). .

Compound (54) [α] ²⁴ _D + 22.3 ° (C 0.783, MeOH) IR (CHCl ₃ ) cm ⁻¹ : 1750, 1721, 1389, 1369, 1080, 1040. ¹ H-NMR (CDCl ₃ , δppm): 5.83 (1H, dd, J = 10.0Hz, J = 9.0Hz, H-3 ′), 5.45 (1H, d, J = 8.6Hz, H-1 ′), 5.19 (1H, t, J = 9.6Hz, H-4 ′), 3.6 (3H, s, OMe), 2.30 (2H, t, J = 7.5Hz, -CH ₂ CO-), 2.09, 2.04, 1.87 (each 3H, each s, 3 × OAc), 1.7 ~ 1.1 (12H, m, -CH ₂ ) _6-

Elemental analysis C ₅₀ H ₆₁ NO ₁₇ · H ₂ O Calculated value C 62.11% H 6.57% N 1.45% Actual value C 61.87% H 6.27% N 1.50%

Compound (55) [α] ²⁵ _D + 16.0 ° (C 0.688, CHCl ₃ ) ¹ H-NMR (CDCl ₃ , δppm): 5.78 (1H, dd, J = 9.2Hz, 11Hz, H-3 ′ or H-3 "), 5.76 (1H, dd, J = 9.2Hz, 11Hz, H-3" or H-3 '), 5.47 (1H, d, J = 8Hz, H-1' or H-1 " ), 5.35 (1H, d, J = 8.4Hz, H-1 "or H-1 '), 5.14 (1H, t, J = 9.4Hz, H-4' or H-4"), 5.13 (1H, t, J = 9.6Hz, H-4 "or H-4 '), 3.68 (3H, s, OMe), 2.05, 2.01, 1.99, 1.84, 1.82 (total 18H, 6 × Ac).

Elemental analysis C ₇₀ H ₈₀ N ₂ O ₂₆・ H ₂ O Calculated value C 60.77% H 5.97% N 2.03% Measured value C 60.63% H 5.85% N 1.96%

Reference Example 25 8-Methoxycarbonyloctyl 6-O- (2-deoxy-2-phthalimido-β-D-glucopyranosyl)
-2,4-Di-O-benzyl-α-D-mannopyranoside (56)

In the same manner as in Reference Example 6, 161 mg (0.176 mmol) of compound (54) was de-O-acetylated in the presence of a catalytic amount of sodium methoxide in methanol to obtain 111 mg of compound (56). .

[Α] ²⁴ _D +5.5 ° (C 0.667, CHCl ₃ ) ¹ H-NMR (CDCl ₃ , δppm): 7.72 to 7.15 (14H, m, aromatic H), 5.31 (1H, d, J = 8.1 Hz, H-1 ′), 3.65 (3H, s, OMe), 2.30 (2H, t, J = 7.5Hz, -C H ₂ C O-), 1.7 ~ 1.1 (12H, m,-(CH ₂ ) _6- )

Reference Example 26 8-Methoxycarbonyloctyl 6-O- (2-acetamido-2-deoxy-β-D-glucopyranosyl)
-2,4-di-O-benzyl-α-D-mannopyranoside (57)

In the same manner as in Reference Example 23, 104 mg (0.132 mmol) of the compound (56) was heated to reflux with hydrazine acetate in methanol to dephthaloylate and then N-acetylated with acetic anhydride to give the compound (57). ) 66
to obtain mg.

[Α] ²⁵ _D + 12.3 ° (C 0.733, MeOH) IR (KBr) cm ⁻¹ : 1740, 1652, 1555 ¹ H-NMR (CD ₃ OD, δppm): 7.25 to 7.48 (10H, m, aromatic H), 4.69 (1H, d, J = 1.6Hz, H-1), 4.47 (1H, d, J = 8.2Hz, H-1 '), 3.63 (3H, s, OMe), 2.30 (2H, t, J = 7.5Hz, -C H ₂ C O-), 1.98, 1.86 (total 3H, NAc), 1.7 ~ 1.2 (12H, m,-(CH ₂ ) _6- )

Example 17 8-Methoxycarbonyloctyl 6-O- (2-acetamido-2-deoxy-β-D-glucopyranosyl)
-Α-D-Mannopyranoside (58) 54 mg (0.0826 mmol) of compound (57) was hydrogenated in the presence of 10% palladium on carbon in methanol and de-O-benzylated to give 36 mg of compound (58). .

[Α] ²⁴ _D -12.4 ° (C 0.723, MeOH) ¹ H-NMR (CD ₃ OD, δppm): 4.68 (1H, d, J = 1.2Hz, H-1), 4.44 (1H, d , J = 8.4Hz, H-1 ′), 3.65 (3H, s, OMe), 2.23 (2H, t, J = 7.5Hz, -CH ₂ C O-), 1.98 (3H, s, NAc), 1.7 ~ 1.25 (12H, m,-(CH ₂ ) _6- )

Reference Example 27 8-Methoxycarbonyloctyl 3,6-di-O-
(2-Deoxy-2-phthalimido-β-D-glucopyranosyl) -2,4-di-O-benzyl-α-D-mannopyranoside (59)

In the same manner as in Reference Example 6, 1.0 g (0.732 mmol) of the crude product (55) was de-O-acetylated in the presence of a catalytic amount of sodium methoxide in methanol to give compound (59). 277 mg were obtained.

[Α] ²⁵ _D + 2.1 ° (C 0.973, MeOH) IR (KBr) cm ⁻¹ : 1775, 1712, 1390, 1072. ¹ H-NMR (CD ₃ OD, δppm): 5.34 (1H, d , J = 8.2Hz, H-1 'or H-1 "), 5.21 (1H, d, J = 8.4Hz, H-1" or H-1'), 3.65 (3H, s, OMe), 2.35 ( 2H, t, J = 7.5Hz, -C H ₂ C O-), 1.7 ~ 1.1 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₅₈ H ₆₈ N ₂ O ₂₀ H ₂ O Calculated value C 61.58% H 6.24% N 2.48% Actual value C 61.70% H 6.23% N 2.66%

Reference Example 28 8-Methoxycarbonyloctyl 3,6-di-O-
(3,4,6-Tri-O-acetyl-2-acetamido-2-deoxy-β-D-glucopyranosyl) -2,4
-Di-O-benzyl-α-D-mannopyranoside (6
0)

3.06 g (33.28 mmol) of hydrazine acetate was added to 50 ml of a methanol solution of 277 mg (0.256 mmol) of compound (59), and the mixture was heated and stirred under a nitrogen stream for 3 hours. After cooling to room temperature, the reaction solution was concentrated to dryness under reduced pressure to obtain 3.59 g of a residue. The residue was dissolved in 15 ml of pyridine, 15 ml of acetic anhydride was added, and the mixture was left at room temperature for 48 hours. The reaction mixture was poured into ice water and the product was extracted with ethyl acetate. The ethyl acetate phase,
It was washed successively with dilute hydrochloric acid, an aqueous sodium hydrogen carbonate solution and water. After drying over magnesium sulfate, the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate) to obtain 212 mg of the compound (60) (yield 71.6%).

[Α] ²⁵ _D + 1.3 ° (C 1.085, CHCl ₃ ) ¹ H-NMR (CDCl ₃ , δppm): 7.5 to 7.2 (10H, m, aromatic H), 3.67 (3H, s, OMe), 2.42, 2.16, 2.05, 2.02, 2.00, 1.84, 1.75, 1.72 (each 3H, each s, NAc and OAc), 2.30 (2H, t, J = 7.5Hz, -C H ₂ CO-), 1.7 ~ 1.2 ( 12H, m,-(CH ₂ ) _6- )

Reference Example 29 8-Methoxycarbonyloctyl 3,6-di-O-
(2-acetamido-2-deoxy-β-D-glucopyranosyl) -2,4-di-O-benzyl-α-D-mannopyranoside (61)

In the same manner as in Reference Example 6, 190 mg (0.164 mmol) of compound (60) was de-O-acetylated using a catalytic amount of sodium methoxide in methanol to obtain 134 mg of compound (61).

[Α] ²⁵ _D + 3.4 ° (C 0.698, MeOH) IR (KBr) cm ⁻¹ : 1735, 1655, 1555, 1070. ¹ H-NMR (CD ₃ OD, δppm): 7.6 to 7.25 (10H , m, aromatic H), 3.64 (3H, s, OMe), 2.30 (2H, t, J = 7.5Hz, -CH ₂ CO-), 1.98 (3H, s, NAc), 1.88, 1.76 (total 3H, NAc), 1.7 to 1.2 (12H, m,-(CH ₂ ) _6- )

Elemental analysis C ₄₆ H ₆₈ N ₂ O ₁₈・ 2.2H ₂ O Calculated value C 56.57% H 7.47% N 2.87% Actual value C 56.87% H 7.24% N 2.70%

Example 18 8-Methoxycarbonyloctyl 3,6-di-O-
(2-Acetamido-2-deoxy-β-D-glucopyranosyl) -α-D-mannopyranoside (62) 143 mg of compound (61) was hydrogenated in methanol in the presence of 10% palladium carbon to de-O-benzylate. 68 mg of compound (62) was obtained.

[Α] ²⁸ _D +6.1 ° (C 0.23, MeOH) ¹ H-NMR (CD ₃ OD, δppm): 4.70 (1H, d, J = 1.5Hz, H-1), 4.56 (1H, d , J = 8.5Hz, H-1 ′ or H-1 "), 4.46 (1H, d, J = 8.5Hz, H-1" or H-1 '), 3.65 (3H, s, OMe), 2.32 ( 2H, t, J = 7.5Hz, -C H ₂ C O-), 1.98 (3H, s, NAc), 2.00, 1.89 (total 3H, s, NAc), 1.65 ~ 1.30 (12H, m,-(CH ₂ ) _6- )

[0211] Elemental analysis _{C 32 H 56 N 2 O 18} · 3H 2 O Calculated C 47.40% H 7.71% N 3.46 % Found C 47.12% H 7.55% N 3.13 %

Reference Example 30 8-Methoxycarbonyloctyl 2,3,4-tri-
O-benzyl-α-L-fucopyranoside (65) and 8-methoxycarbonyloctyl 2,3,4-tri-
O-benzyl-β-L-fucopyranoside (67)

2,3,4-tri-O-benzyl-1-p
-Nitrobenzoyl-β-L-fucopyranose 4.00
New 2,3,4 prepared from g (6.85 mmol)
-Tri-O-benzyl-α-L-fucopyranosyl bromide (63) [Carbohydr, Res., 18, 219-226 (1971).]
To a 40 ml solution of benzene in the above solution, 1.29 g (6.85 mmol) of 8-methoxycarbonyloctanol, 1.73 g (6.85 mmol) of mercuric cyanide and 3.60 g of powdered anhydrous calcium sulfate were added, and the mixture was stirred at room temperature in an argon stream. And stirred for 2 days. After removing the precipitate by filtration, 60 ml of ethyl acetate was added to the filtrate, and a saturated aqueous solution of sodium hydrogencarbonate 60 m was added.
1 and 60 ml of water were sequentially washed. The syrup obtained by dehydration over anhydrous magnesium sulfate and concentration under reduced pressure was subjected to silica gel column chromatography (n-hexane: ethyl acetate = 1).
The compound (65) 1.66 g (yield 4
0.1%) and the compound (67) (yield 12.6%) were obtained.

Compound (65) [α] ²⁴ _D −34.0 ° (C 1.0, CHCl ₃ ) IR (NaCl) cm ⁻¹ : 1725 ¹ H-NMR (CDCl ₃ , δppm): 1.10 (3H, d, J = 7.5Hz, H-6, CH ₃ ), 2.28 (2H, t, J = 7.6Hz, C H ₂ C OOCH ₃ ), 3.65 (3H, s, OCH ₃ )

Compound (67) [α] ²⁴ _D + 6.2 ° (C 1.0, CHCl ₃ ) IR (NaCl) cm ⁻¹ : 1725 ¹ H-NMR (CDCl ₃ , δppm): 1.15 (3H, d, J = 7.5Hz, H-6), 2.28 (2H, t, J = 7.6Hz, C H ₂ C OOCH ₃ ), 3.65 (3H, s, OCH ₃ ), 4.32 (1H, d, J = 8.6Hz, H- 1)

Example 19 8-Methoxycarbonyloctyl α-L-fucopyranoside (66) To a solution of 1.89 g (3.12 mmol) of the compound (65) in 50 ml of acetic acid was added 1.0 g of 10% palladium carbon, and Parr's apparatus ( 50 psi) at room temperature for 3 days. After removing the precipitate by filtration, the solid obtained by concentrating the filtrate under reduced pressure was purified by silica gel column chromatography (chloroform: methanol = 30: 1) to give compound (66).
848 mg (yield 81.3%) was obtained.

[Α] ²³ _D -110.1 ° (C 1.0, MeOH) IR (NaCl) cm ⁻¹ : 3350, 1740 ¹ H-NMR (CD ₃ OD, δppm): 1.18 (3H, d, J = 7.5Hz , H-6,), 2.31 (2H, t, J = 7.6Hz, C H ₂ C OOCH ₃ ), 3.65 (3H, s, OCH ₃ ), 4.74 (1H, d, J = 2.7Hz, H-1 )

Example 20 8-Methoxycarbonyloctyl β-L-fucopyranoside (68) Compound (67) (596 mg, 0.985 mmol) was added to Example 2.
226 mg of compound (68) by catalytic reduction in the same manner as
(Yield 68.6%) was obtained.

[Α] ²³ _D + 4.5 ° (C 1.0, MeOH) IR (NaCl) cm ⁻¹ : 3350, 1740 ¹ H-NMR (CD ₃ OD, δppm): 1.21 (3H, d, J = 7.5Hz , H-6,), 2.31 (2H, t, J = 7.6Hz, C H ₂ C OOCH ₃ ), 3.65 (3H, s, OCH ₃ ), 4.17 (1H, d, J = 8.1Hz, H-1 )

Reference Example 31 6-Methoxycarbonylhexyl 2,3,4-tri-
O-acetyl-β-L-fucopyranoside (69)

1,2,3,4-tetra-O-acetyl-
New 2,3,4-tri-O-acetyl-α prepared from 1.00 g (3.01 mmol) of L-fucopyranose
-L-fucopyranosyl bromide (64) [Carbohydr,
Res., 4, 189-195 (1967)] in 10 ml of benzene,
241 mg of 6-methoxycarbonylhexanol (1.5
0 mmol), 380 mg (1.50 mmol) of mercuric cyanide and 900 mg of anhydrous powdery calcium sulfate were added, and the mixture was stirred at room temperature in an argon stream for 24 hours. After removing the precipitate by filtration, 15 ml of ethyl acetate was added to the filtrate, and the mixture was washed successively with 15 ml of saturated aqueous sodium hydrogen carbonate solution and 15 ml of water.
The syrup obtained by dehydration over anhydrous magnesium sulfate and concentration under reduced pressure was purified by silica gel column chromatography (toluene: ethyl acetate = 8: 1) to give compound (69) 4.
01 mg (yield 61.6%) was obtained.

[Α] ²³ _D +1.7 ° (C 1.0, CHCl ₃ ) IR (NaCl) cm ⁻¹ : 1740 ¹ H-NMR (CDCl ₃ , δppm): 1.22 (3H, d, J = 6.4Hz, H -6), 1.99, 2.05, 2.17 (3H, s, CH ₃ CO), 2.31 (2H, t, H = 7.6Hz, C H ₂ C OOCH ₃ ), 3.67 (3H, s, OCH ₃ ), 4.42 ( 1H, d, J = 8.1Hz, H-1)

Example 21 6-Methoxycarbonylhexyl β-L-fucopyranoside (70) Compound (69) 389 mg (0.899 mmol) in methanol 4 ml solution 28% sodium methylate 0.1 ml
Was added and the mixture was stirred at room temperature for 2 hours. Amber Light IR-
After neutralization with 120B (H ⁺ type), the resin was filtered off and the filtrate was concentrated under reduced pressure to obtain 268 mg (yield 97.3%) of syrupy compound (70).

[Α] ²³ _D +3.3 ° (C 0.64, MeOH) IR (NaCl) cm ⁻¹ : 3450, 1740 ¹ H-NMR (CD ₃ OD, δppm): 1.23 (3 H, d, J = 6.6 Hz) , H-6,), 2.37 (2H, t, J = 7.6Hz, CH ₂ C OOCH ₃ ), 3.67 (3H, s, OCH ₃ ), 4.19 (1H, d, J = 7.3Hz, H-1)

Reference Example 32 8-Methoxycarbonyloctyl 2-acetamido-
3-O- (2,3,4-tri-O-acetyl-β-L-
Fucopyranosyl) -4,6-O-benzylidene-2-deoxy-β-D-glucopyranoside (72)

1,2,3,4-tetra-O-acetyl-
New 2,3,4-tri-O-acetyl-α prepared from 156 mg (0.469 mmol) L-fucopyranose
-Benzene of L-fucopyranosyl bromide (64)-
8-Methoxycarbonyloctyl 2-acetamido-4,6-O- was added to a nitromethane (10 ml-10 ml) solution.
Benzylidene-2-deoxy-β-D-glucopyranoside (71) [J. Am. Chem. Soc., 97, 4076-4083 (197)
5)] 150 mg (0.313 mmol) and mercuric cyanide 79 mg (0.313 mmol) were added, and the mixture was stirred at room temperature in an argon stream for 24 hours. After filtering off the precipitate,
10 ml of ethyl acetate was added to the filtrate, and the mixture was washed successively with 10 ml of saturated aqueous sodium hydrogen carbonate solution and 10 ml of water. The syrup obtained by dehydration over anhydrous magnesium sulfate and concentration under reduced pressure was purified by silica gel column chromatography (chloroform: methanol = 150: 1) to obtain a powdery compound (7
2) 205 mg (yield 87.1%) was obtained.

[Α] ²⁷ _D -15.2 ° (C 1.1, CHCl ₃ ) IR (KBr) cm ^-1 : 1750, 1230, 1080 ¹ H-NMR (CDCl ₃ , δppm): 1.18 (3H, d, J = 5.9Hz, H-6 ′), 1.81 (3H, s, NCOCH ₃ ), 1.97, 1.98, 2.16 (3H, s, OCOCH ₃ ), 2.30 (2H, t, J = 7.1Hz, CH ₂ C OOCH ₃ ) , 3.67 (3H, s, OCH ₃ ), 4.67 (1H, d, J = 7.8Hz, H-1), 5.86 (1H, d, J = 6.8Hz, NH)

Example 22 8-Methoxycarbonyloctyl 2-acetamido-
2-deoxy-3-O- (β-L-fucopyranosyl)-
β-D-glucopyranoside (73) To a solution of 124 mg (0.165 mmol) of compound (72) in 10 ml of dioxane, 3.3 ml of 0.5M sulfuric acid was added, and 90
Stir at 20 ° C. for 20 minutes. Chloroform (20 ml) was added, and the mixture was washed successively with saturated aqueous sodium hydrogen carbonate solution (20 ml) and water (20 ml). The residue obtained by dehydration over anhydrous magnesium sulfate and concentration under reduced pressure was dissolved in 2 ml of methanol, 0.02 ml of 28% sodium methylate was added, and the mixture was stirred at room temperature for 2 hours.
After neutralization with Amberlite IR-120B (H ⁺ type), the resin was filtered off, and the syrup obtained by concentrating the filtrate under reduced pressure was subjected to silica gel column chromatography (chloroform: methanol: water = 16: 4: 1). Compound (73) 3 after purification
3.5 mg (yield 47.0%) was obtained.

[Α] ²⁵ _D -26.9 ° (C 0.82, MeOH) IR (KBr) cm ^-1 : 3400, 1740, 1660, 1080 ¹ H-NMR (CD ₃ OD, δppm): 1.27 (3H, d, J = 6.4Hz, H-6 ′), 1.95 (3H, s, NCOCH ₃ ), 2.30 (2H, t, J = 7.6Hz, CH ₂ COOCH ₃ ), 3.65 (3H, s, OCH ₃ ), 4.39 ( 1H, d, J = 7.3Hz, H-1 ′), 4.51 (1H, d, J = 8.5Hz, H-1)

Reference Example 33 8-Methoxycarbonyloctyl 2-acetamido-
3,4-di-O-acetyl-2-deoxy-6-O-trityl-β-D-glucopyranoside (74)

8-Methoxycarbonyloctyl 2-acetamido-2-deoxy-β-D-glucopyranoside [J. Am. Chem. Soc., 97, 4076-4083 (1975)] 895
About 5 ml of a 20 ml solution of mg (2.29 mmol) in pyridine
It was concentrated under reduced pressure until. Trityl chloride 715 mg (2.
(56 mmol) was added, and the mixture was stirred at room temperature overnight. Water 0.1
After adding ml, the mixture was stirred at room temperature for 10 minutes and then concentrated under reduced pressure. Chloroform (30 ml) was added to the residue, and the mixture was washed successively with saturated sodium hydrogen carbonate (30 ml) and water (30 ml). After dehydration over anhydrous magnesium sulfate, the mixture was concentrated under reduced pressure to obtain a syrupy compound. Add 5 ml of pyridine and 5 ml of acetic anhydride, and add 4 at room temperature.
Stirred for hours. 2 ml of methanol was added under ice cooling and the mixture was stirred for 20 minutes. Chloroform (30 ml) was added to the residue obtained by concentration under reduced pressure, and the mixture was washed successively with saturated sodium hydrogencarbonate (30 ml) and water (30 ml). The syrup obtained by dehydration over anhydrous magnesium sulfate and concentration under reduced pressure was purified by silica gel column chromatography (chloroform: methanol = 120: 1) to obtain 1.12 g of compound (74) (yield 68.3%). .

Mp 159 to 160 ° C. [α] ²⁵ _D + 31.8 ° (C 0.86, CHCl ₃ ) IR (KBr) cm ⁻¹ : 1740 ¹ H-NMR (CDCl ₃ , δppm): 1.71 (3H, s, NCOCH ₃ ), 1.94, 2.00 (3H, s, OCOCH ₃ ), 2.27 (2H, t, J = 7.3Hz, CH ₂ COOCH ₃ ), 3.65 (3H, s, OCH ₃ ), 4.61 (1H, d, J = 8.3Hz, H-1), 5.51 (1H, d, J = 8.8Hz, NH), 7.2 ~7.3 and 7.4 ~7.5 (15H, m, C 6 H 5)

Reference Example 34 8-Methoxycarbonyloctyl 2-acetamido-
3,4-Di-O-acetyl-2-deoxy-β-D-glucopyranoside (75) To a solution of 1.00 g (1.39 mmol) of compound (74) in 5 ml of acetic acid was added 25% hydrogen bromide at 15 ° C. 1 ml of acetic acid solution was added and stirred for 1.5 minutes. After removing the precipitate by filtration, the filtrate was added to 30 ml of ice water and extracted 3 times with 10 ml of chloroform. The chloroform solution was washed with 30 ml of water, dehydrated over anhydrous magnesium sulfate, and concentrated under reduced pressure to obtain a syrup, which was subjected to silica gel column chromatography (chloroform:
Compound (75) 49 after purification with methanol = 40: 1)
3 mg (74.6% yield) was obtained.

Mp 129 to 130 ° C. [α] ²⁵ _D -11.2 ° (C 0.98, CHCl ₃ ) IR (KBr) cm ⁻¹ : 1740 ¹ H-NMR (CDCl ₃ , δppm): 1.94 (3H, s, NCOCH ₃ ), 2.04, 2.05 (3H, s, OCOCH ₃ ), 2.30 (2H, t, J = 7.6Hz, CH ₂ C OOCH ₃ ), 3.67 (3H, s, OCH ₃ ), 3.7 ~ 3.9 (4H, m , H-2, H-5, H-6), 4.69 (1H, d, J = 8.3Hz, H-1), 5.00 (1H, t, J = 9.8Hz, H-4), 5.32 (1H, t, J = 9.6Hz, H-3), 5.54 (1H, d, J = 8.6Hz, NH)

Reference Example 35 8-methoxycarbonyloctyl 2-deoxy-3,
4-di-O-acetyl-6-O- (2,3,4-tri-
O-acetyl-β-L-fucopyranosyl) -2-deoxy-β-D-glucopyranoside (76)

1,2,3,4-tetra-O-acetyl-
New 2,3,4-tri-O-acetyl-α prepared from 126 mg (0.379 mmol) L-fucopyranose
120 mg of the compound (75) (0. 0%) in 8 ml of 1,2-dichloroethane solution of -L-fucopyranosyl bromide (64).
252 mmol), silver triflate 227 mg (0.88)
2 mmol) and 176 mg (1.51 mmol) of 1,1,3,3-tetramethylurea were added, and the mixture was stirred at room temperature in an argon stream for 24 hours. After removing the precipitate by filtration, 10 ml of chloroform was added to the filtrate and the mixture was washed with 10 ml of water. Syrup obtained by dehydration over anhydrous magnesium sulfate and concentration under reduced pressure was subjected to silica gel column chromatography (chloroform:
Compound (76) 1 purified by methanol = 200: 1)
36 mg (yield 72.2%) was obtained.

[Α] ²⁷ _D + 4.5 ° (C 0.81, CHCl ₃ ) IR (NaCl) cm ⁻¹ : 1740 ¹ H-NMR (CDCl ₃ , δppm): 1.19 (3H, d, J = 6.4Hz, H -6 ′), 1.92, 1.96. 1.99, 2.00, 2.10, 2.15 (3H, s, COCH ₃ ), 2.29 (2H, t, J = 7.6H z, CH ₂ C OOCH ₃ ), 5.49 (1H, d, J = 8.8Hz, NH),

Example 23 8-Methoxycarbonyloctyl 2-acetamido-
2-deoxy-6-O- (β-L-fucopyranosyl)-
β-D-Glucopyranoside (77) 28% sodium methylate 0.04 in a solution of 125 mg (0.167 mmol) of compound (76) in 2 ml of methanol.
ml was added, and the mixture was stirred at room temperature overnight. Amber Light IR-
After neutralization with 120B (H ⁺ type), the resin was filtered off, and the syrup obtained by concentrating the filtrate under reduced pressure was subjected to silica gel column chromatography (chloroform: methanol: water = 16:
4: 1) and then 68.9 mg (76.
7%).

[Α] ²⁷ _D -16.7 ° (C 0.84, MeOH) IR (KBr) cm ^-1 : 3400, 1740, 1080 ¹ H-NMR (CD ₃ OD, δppm): 1.26 (3H, d, J = 6.6Hz, H-6 ′), 1.95 (3H, s, NCOCH ₃ ), 2.30 (2H, t, J = 7.6Hz, CH ₂ COOCH ₃ ), 3.7 ~ 3.8 (2H, m, H-6), 4.24 (1H, d, J = 7.3Hz H-1 ′), 4.40 (1H, d, J = 8.3Hz, H-1)

Reference Example 36 2,3,4-tri-O-acetyl-6-0- (2,3,3
4,6-Tetra-O-acetyl-α-D-galactopyranosyl) -α-D-glucopyranosyl bromide (acetobromomelibiose) (78)

12.5 g of melibiose (manufactured by Sigma)
(36.5 mmol), pyridine (130 ml) and acetic anhydride (38 ml) were stirred and reacted at room temperature for 24 hours.
The solvent was distilled off under reduced pressure to obtain peracetyl melibiose. The peracetylmelibiose obtained was dissolved in 50 ml of acetic acid, 20 ml of 25% hydrogen bromide-acetic acid solution was added, and the mixture was stirred at room temperature for 2
Stir for 4 hours. After completion of the reaction, 200 ml of chloroform was added, the reaction mixture was poured into 300 ml of ice water, stirred well, transferred to a separating funnel, and the chloroform phase was separated, and washed successively with water, saturated aqueous sodium hydrogen carbonate and saturated brine. After the obtained chloroform solution was dried over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure to obtain crude bromide. Silica gel column chromatography (ethyl acetate: hexane = 1: 1)
To give 14.7 g (yield 60%) of colorless acetambromomelobiose (78) in the form of a colorless caramel.

[Α] ²⁰ _D +178.5 ° (C 1.27, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 1750 (OAc) ¹ H-NMR (CDCl ₃ , δppm): 1.99 (3H, s, OAc) , 2.04 (3H, s, OAc), 2.05 (3H, s, OAc), 2.07 (3H, s, OAc), 2.10 (3H, s, OAc), 2.12 (3H, s, OAc), 2.14 (3H, s, OAc), 3.52 ~ 3.82 (2H, m), 4.00 ~ 4.36 (4H, m), 4.79 (1H, dd), 5.00 ~ 5.20 (3H, m), 5.34 (1H, dd), 5.47 (1H, dd), 5.56 (1H, t), 6.59 (1H, d, J = 3.9Hz, H-1)

Reference Example 37 3,4-di-O-acetyl-6-O- (2,3,4,6
-Tetra-O-acetyl-α-D-galactopyranosyl) glucal (79) Acetobromomelibiose (78) 4 g (5.7 mmol), a solution of acetic acid 25 ml and water 25 ml was cooled to 0-5 ° C. Below, 8 g of zinc powder was added, and the mixture was stirred and reacted for 4 hours. After completion of the reaction, insoluble matter was removed by filtration through Celite, and the filtrate was poured into water and the precipitate was extracted with ethyl acetate. The ethyl acetate phase was separated, washed with water repeatedly, washed with saturated brine, and then dried over anhydrous magnesium sulfate. The crude product (2.7 g) obtained after the solvent was distilled off was purified by flash chromatography (ethyl acetate: hexane = 1: 2) to give 1.32 g.
Glycal (79) was obtained (yield 41.3%).

Mp 106-107 ° C. [α] ²⁰ _D + 83.8 ° (C 1.00, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 1740 (OAc), 1650 (C = C) ¹ H-NMR (CDCl ₃ , δppm): 1.99 (3H, s, OAc), 2.05 (6H, s, OAc x 2), 2.10 (3H, s, OAc), 2.11 (3H, s, OAc), 2.14 (3H, s, OAc), 3.60 ~ 3.92 (2H, m), 4.00 ~ 4.12 (2H, m), 4.16 ~ 4.32 (2H, m), 4.80 ~ 4.88 (1H, q), 5.00 ~ 5.40 (5H, m), 5.47 (1H, d , J = 3.4Hz, H-1 '), 6.48 (1H, d, J = 7.3Hz, -OC H = CH-)

Reference Example 38 1,3,4-tri-O-acetyl-6-O- (2,3,3
4,6-Tetra-O-acetyl-α-D-galactopyranosyl) -2-azido-2-deoxy-α-D-glucopyranose (80) and 1,3,4-tri-O-acetyl -6-O- (2,3,4,6-tetra-O-acetyl-α-D-galactopyranosyl) -2-azido-2-deoxy-α-D-mannopyranose (82)

6.8 g (12.2 mmol) of glycal (79) was dissolved in 70 ml of acetonitrile and this was added to a mixture of 1.2 g (18.8 mmol) of sodium azide and 20 g (36.5 mmol) of ammonium cerium nitrate. The mixture was added dropwise at -20 ° C and vigorously stirred. After reacting for 5 hours, 100 ml of ether and 50 ml of water were added to the reaction solution and shaken well, the ether phase was separated, and washed successively with water and saturated brine. Crude product obtained after distilling off ether 7.
48 g flash chromatography (ethyl acetate:
Separation and purification with hexane = 3: 2), 4.15 g (51.
5%) of the azide compound was obtained. This was dissolved in 25 ml of acetic acid, 1.23 g of sodium acetate was added, and the mixture was reacted at 100 ° C for 2 hours. After completion of the reaction, 300 ml of methylene chloride and 30 ml of water were added and shaken well, the methylene chloride phase was separated, and washed successively with saturated aqueous sodium hydrogen carbonate and saturated brine. After drying over magnesium sulfate, the solvent was distilled off under reduced pressure and the obtained crude product was purified by flash chromatography (ethyl acetate: hexane = 1: 1) to give compound (80).
To give 2.35 g (yield 49.2%) of Compound (82) and 1.55 g (yield 32.4%), respectively.

Compound (80) [α] ²⁰ _D +145.2 ° (C 1.00, CHCl ₃ ) IR (Nujol) cm ⁻¹ : 2120 (N ₃ ), 1750 (OAc) ¹ H-NMR (CDCl ₃ , δppm) : 1.99 (3H, s, OAc), 2.05 (3H, s, OAc), 2.06 (3H, s, OAc), 2.10 (3H, s, OAc), 2.11 (3H, s, OAc), 2.14 (3H, s, OAc), 2.21 (3H, s, OAc), 3.50 ~ 3.80 (3H, m), 3.92 ~ 4.24 (4H, m), 4.96 ~ 5.20 (3H, m), 5.24 ~ 5.56 (3H, m), 6.25 (1H, d, J = 3.9Hz, H-1)

Compound (82) (α) ²⁰ _D +135.8 ° (C 0.66, CHCl ₃ ) IR (Nujol) cm ⁻¹ : 2120 (N ₃ ), 1750 (OAc) ¹ H-NMR (CDCl ₃ , δppm) : 1.99 (3H, s, OAc), 2.06 (3H, s, OAc), 2.07 (3H, s, OAc), 2.12 (6H, s, OAc x 2), 2.14 (3H, s, OAc), 2.19 ( 3H, s, OAc), 3.50 ~ 3.80 (2H, m), 3.88 ~ 4.32 (5H, m), 5.00 ~ 5.20 (2H, m), 5.20 ~ 5.40 (3H, m), 5.45 (1H, d, J = 3.4Hz, H-1 ′), 6.04 (1H, d, J = 2.0Hz, H-1)

Reference Example 39 2-Azido-2-deoxy-3,4-di-O-acetyl-6-O- (2,3,4,6-tetra-O-acetyl-
α-D-galactopyranosyl) -α-D-glucopyranosyl bromide (81)

Titanium tetrabromide (1.92 g, 5.2 mmol) was added to a solution of the compound (80) (2.3 g, 3.47 mmol), methylene chloride (35 ml) and ethyl acetate (3.5 ml), and the mixture was stirred at room temperature for 24 hours. It was made to react. After completion of the reaction, the reaction solution was transferred to a funnel, 100 ml of methylene chloride and 20 ml of saturated aqueous sodium hydrogen carbonate were added and shaken well, and the methylene chloride phase was separated. The organic layer was washed with water and saturated brine in that order, dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure to obtain 2.37 g of a crude product. This was purified by flash chromatography (silica gel 40 g, solvent system, ethyl acetate: hexane = 3: 2) to obtain 1.2 g of colorless caramel bromide (81).
(Yield 50.6%) was obtained.

[Α] ²⁰ _D +191.6 ° (C 0.64, CHCl ₃ ) IR (Nujol) cm ⁻¹ : 2120 (N ₃ ), 1750 (OAc) ¹ H-NMR (CDCl ₃ , δppm): 1.99 (3H , s, OAc), 2.05 (3H, s, OAc), 2.07 (3H, s, OAc), 2.10 (3H, s, OAc), 2.12 (3H, s, OAc), 2.14 (3H, s, OAc) , 3.52 ~ 3.80 (3H, m), 3.92 ~ 4.32 (4H, m), 4.96 ~ 5.20 (3H, m), 5.24 ~ 5.52 (3H, m), 6.39 (1H, d, J = 3.4Hz, H- 1)

Reference Example 40 3,4-di-O-acetyl-6-O- (2,3,4,6
-Tetra-O-acetyl-α-D-galactopyranosyl) -2-azido-2-deoxy-α-D-mannopyranosyl bromide (83)

According to the method described in Reference Example 39, a solution of 1.5 g (2.27 mmol) of the compound (82), 25 ml of methylene chloride and 2.5 ml of ethyl acetate was added with 1.25 g of titanium tetrabromide (3.4 mmol). B) (83) was obtained as a clear oil. This was crystallized in time, washed with ethyl acetate-hexane and then dried to obtain 1.1 g (yield 71%) of compound (83) as colorless crystals.

Mp146-148 ° C. (decomposition), [α] ²⁰ _D + 201.4 ° (C 0.86, CHCl ₃ ) IR (Nujol) cm ⁻¹ : 2120 (N ₃ ), 1750 (OAc) ¹ H-NMR (CDCl ₃ , δppm): 1.99 (3H, s, OAc), 2.06 (3H, s, OAc), 2.09 (3H, s, OAc), 2.11 (3H, s, OAc), 2.13 (3H, s, OAc), 2.14 (3H, s, OAc), 3.52 ~ 3.80 (2H, m), 3.96 ~ 4.24 (4H, m), 4.32 (1H, q), 5.04 ~ 5.20 (2H, m), 5.24 ~ 5.48 (3H, m ), 5.704 (1H, q), 6.33 (1H, d, J = 1.0Hz, H-1)

Reference Example 41 8-Methoxycarbonyloctyl 3,4-di-O-acetyl-6-O- (2,3,4,6-tetra-O-acetyl-α-D-galactopyranosyl)- 2-azido-2
-Deoxy-β-D-glucopyranoside (86)

8-Methoxycarbonyloctanol 38
0 mg (2.0 mmol), drylite 400 mg, silver carbonate 310 mg (1.12 mmol) and chloroform 7
The ml mixture was stirred at room temperature under an argon stream for 1 hour.
Then a solution of 682 mg (1.0 mmol) of bromide (81) in 3 ml of chloroform was slowly added dropwise, after which 4
Stirred for hours. After the reaction was completed, the mixture was filtered through Celite, and the filtrate was distilled off under reduced pressure to obtain 1.07 g of a transparent oily substance. This is flash chromatographed (ethyl acetate: hexane =
It was purified by 2: 3) to obtain 478 mg (yield 60.5%) of the compound (86).

[Α] ²⁰ _D +103.2 ° (C 0.62, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 2120 (N ₃ ), 1750 (OAc, CO ₂ CH ₃ ) ¹ H-NMR (CDCl ₃ , δppm): 1.20 ~ 1.44 (8H, broad, s), 1.50 ~ 1.72 (6H, broad, m), 1.99 (3H, s, OAc), 2.03 (3H, s, OAc), 2.05 (3H, s, OAc ), 2.08 (3H, s, OAc), 2.11 (3H, s, OAc), 2.24 (3H, s, OAc), 2.30 (2H, t, CH ₂ CO ₂ CH ₃ ), 3.30 ~ 3.80 (5H, m ), 3.67 (3H, s, CO ₂ CH ₃ ) , 3.80 ~ 3.96 (1H, m), 4.00 ~ 4.28 (3H, m), 4.38 (1H, d, J = 7.8Hz, H-1), 4.88 ~ 5.20 (4H, m), 5.24 ~ 5.36 (1H, dd), 5.44 (1H, d, J = 2.4Hz, H-1 ')

Reference Example 42 8-Methoxycarbonyloctyl 3,4-di-O-acetyl-6-O- (2,3,4,6-tetra-O-acetyl-α-D-galactopyranosyl)- 2-azido-2
-Deoxy-α-D-mannopyranoside (84)

By the method described in Reference Example 41, 380 mg (2.0 mmol) of 8-methoxycarbonyloctanol, 400 mg of drylite, 310 mg of silver carbonate (1.1
Bromide (83) 682 mg (1.0 mmol) chloroform 3 in a mixture of 2 mmol) and 7 ml chloroform.
238 mg (yield 30.1
%) Of compound (84).

[Α] ²⁰ _D +141.6 ° (C 0.74, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 2120 (N ₃ ), 1750 (OAc, CO ₂ CH ₃ ) ¹ H-NMR (CDCl ₃ , δppm): 1.20 ~ 1.44 (8H, broad, s), 1.48 ~ 1.70 (6H, broad, m), 1.98 (3H, s, OAc), 2.06 (3H, s, OAc), 2.07 (3H, s, OAc ), 2.09 (3H, s, OAc), 2.12 (3H, s, OAc), 2.14 (3H, s, OAc), 2.31 (2H, t, CH ₂ C O ₂ CH ₃ ), 3.30 ~ 3.80 (3H, m), 3.67 (3H, s, CO ₂ CH ₃ ) , 3.80 ~ 3.92 (1H, m), 3.92 ~ 4.32 (5H, m), 4.76 (1H, d, J = 1.5Hz, H-1), 5.00 ~ 5.48 (6H, m)

Reference Example 43 8-Methoxycarbonyloctyl 3,4-di-O-acetyl-6-O- (2,3,4,6-tetra-O-acetyl-α-D-galactopyranosyl)- 2-acetamido-2-deoxy-β-D-glucopyranoside (87)

To a solution of 450 mg (0.57 mmol) of compound (86), 7 ml of ethyl acetate and 5 ml of acetic anhydride was added 10 parts.
% Palladium-carbon catalyst (400 mg) was added, and the mixture was stirred under a hydrogen stream for 8 hours. After completion of the reaction, the catalyst was removed by filtration, the filtrate was evaporated under reduced pressure, and the residue was dissolved in 10 ml of pyridine and 1 ml of acetic anhydride and stirred for 5 hours. The solution was evaporated under reduced pressure and the obtained crude product was purified by flash chromatography (ethyl acetate: hexane = 3: 2) to give compound (8
125 mg (yield 27.2%) of 7) was obtained.

[Α] ²⁰ _D + 68.3 ° (C 1.00, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 3440 (NH), 1750 (OAc, CO ₂ CH ₃ ), 1680 (NHCOCH ₃ ) ¹ H- NMR (CDCl ₃ , δppm): 1.10 to 1.40 (8H, broad, s), 1.44 to 1.72 (6H, broad, m), 1.94 (3H, s, Ac), 1.99 (3H, s, Ac), 2.03 ( 3H, s, Ac), 2.04 (3H, s, Ac), 2.05 (3H, s, Ac), 2.12 (3H, s, Ac), 2.14 (3H, s, Ac), 2.30 (2H, t, CH ₂ C O ₂ CH ₃ ), 3.30 to 3.90 (5H, m), 3.67 (3H, s, CO ₂ CH ₃ ) , 3.96 to 4.12 (2H, m), 4.16 to 4.32 (1H, m), 4.62 (1H , d, J = 8.3Hz, H-1), 4.90 ~ 5.36 (4H, m), 5.45 (1H, d, J = 2.5Hz,), 5.52 (1H, d, J = 8.8Hz, NH)

Reference Example 44 8-Methoxycarbonyloctyl 3,4-di-O-acetyl-6-O- (2,3,4,6-tetra-O-acetyl-α-D-galactopyranosyl)- 2-acetamido-2-deoxy-α-D-mannopyranoside (85)

According to the method described in Reference Example 43, 200 mg (0.25 mmol) of compound (84) was reduced in the presence of 300 mg of 10% palladium-carbon catalyst to give a colorless caramel compound (85). 86 mg (yield 42.1
%)Obtained.

[Α] ²⁰ _D + 113.14 ° (C 1.59, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 3400 (NH), 1750 (OAc, CO ₂ CH ₃ ), 1680 (NHCOCH ₃ ) ¹ H- NMR (CDCl ₃ , δppm): 1.20 to 1.44 (8H, broad, s), 1.48 to 1.76 (6H, broad, m), 1.98 (3H, s, Ac), 2.00 (3H, s, Ac), 2.05 ( 3H, s, Ac), 2.07 (3H, s, Ac), 2.08 (3H, s, Ac), 2.15 (3H, s, AC), 2.16 (3H, s, Ac), 2.31 (2H, t, J _{= 7.3Hz, CH 2 C O 2} CH 3), 3.30~3.46 (1H, m), 3.50~3.76 (1H, m), 3.67 (3H, s, CO 2 CH 3), 3.80~3.94 (1H, m ), 3.98 ~ 4.32 (3H, m), 4.52 ~ 4.68 (2H, m), 4.92 ~ 5.12 (2H, m), 5.24 ~ 5.50 (4H, m), 6.30 (1H, d, J = 9.3Hz, NH )

Example 24 8-Methoxycarbonyloctyl 2-acetamido-
2-deoxy-6-O- (α-D-galactopyranosyl) -β-D-glucopyranoside (88) A solution of 100 mg (0.124 mmol) of compound (87) in 2 ml of methanol was mixed with 28% sodium methoxide. -Methanol solution (0.05 ml) was added and the reaction was allowed to stir for 21 hours. After completion of the reaction, 3 ml of Amberlite IR-120 resin-methanol was added, stirred for 30 minutes, filtered to remove the resin, and the filtrate was evaporated under reduced pressure to obtain 62.8 mg (yield 91.4
%) Deacetylated product (88) was obtained.

[Α] ²⁰ _D + 47.7 ° (C 1.26, MeOH) IR (CHCl ₃ ) cm ⁻¹ : 3400, 1730 (CO ₂ CH ₃ ), 1650 (NHAc) ¹ H-NMR (CDCl ₃ -CD ₃ OD, δppm): 1.20 ~ 1.40 (8H, broad, s), 1.50 ~ 1.65 (4H, m), 1.98 (3H, s, NHAc), 2.31 (2H, t, CH ₂ C O ₂ CH ₃ ), 3.40 ~ 3.50 (4H, m), 3.60 ~ 3.65 (1H, m), 3.66 (3H, s, CO ₂ CH ₃ ) , 3.68 ~ 3.78 (5H, m), 3.80 ~ 3.90 (2H, m), 3.92 (1H , broad, s), 4.03 (1H, dd), 4.43 (1H, d, J = 8.3Hz, H-1), 4.89 (1H, broad, s, H-1 ')

Example 25 8-Methoxycarbonyloctyl 2-acetamido-
2-Deoxy-6-O- (α-D-galactopyranosyl) -α-D-mannopyranoside (89) Compound (85) 235 was prepared according to the method described in Example 24.
140.3 of deacetylation of mg (0.29 mmol)
mg (86.9%) of deacetylated product (89) was obtained.

[Α] ²⁰ _D + 91.8 ° (C 0.4, MeOH) IR (CHCl ₃ ) cm ⁻¹ : 3400, 1730 (CO ₂ CH ₃ ), 1650 (NHAc) ¹ H-NMR (CDCl ₃ , δppm) : 1,28 ~ 1.40 (8H, broad, s), 1.55 ~ 1.70 (4H, m), 2.03 (3H, s, NHAc), 2.32 (2H, t, J = 7.6Hz, CH ₂ C O ₂ CH ₃ ), 3.36 ~ 3.42 (1H, m), 3.62 ~ 3.69 (2H, m), 3.67 (3H, s, CO ₂ CH ₃ ) , 3.69 ~ 3.79 (4H, m), 3.79 ~ 3.84 (2H, m), 3.90 ~ 3.94 (2H, m), 3.920 (1H, dd), 4.04 (1H, dd), 4.30 (1H, dd), 4.69 (1H, d, J = 1.0Hz, H-1), 4.99 (1H, d, J = 3.2Hz, H-1 ′)

Reference Example 45 8-Methoxycarbonyloctyl 2,3,4-tri-
O-acetyl-6-O- (2,3,4,6-tetra-O
-Acetyl-α-D-galactopyranosyl) -β-D-
Glucopyranoside (90) and 8-methoxycarbonyloctyl 2,3,4-tri-O-acetyl-6-O
-(2,3,4,6-tetra-O-acetyl-α-D-
Galactopyranosyl) -α-D-glucopyranoside (9
2)

8-Methoxycarbonyloctanol 2.
33 g (12.4 mmol), dry light 4.5 g,
A mixture of 3.2 g (12.6 mmol) of mercuric cyanide and 50 ml of anhydrous benzene was stirred under a stream of argon for 1 hour. After that, 5 g of acetobromomellibiose
A solution of (7.15 mmol) in 10 ml of benzene was added dropwise, and the mixture was stirred at room temperature for 4 hours. After the reaction was completed, the insoluble matter was removed by filtration, and the filtrate was distilled off under reduced pressure to give 6.88 g of a crude product.
I got This was purified by flash chromatography (ethyl acetate: hexane = 1: 1) to give compound (90).
2.55 g (yield 44.2%) and compound (92)
0.94 g (yield 16.3%) were obtained.

Compound (90) (α) ²⁰ _D + 53.9 ° (C 1.14, CHCl ₃ ) IR (CHCl ₃ ) cm ^-1 : 750 (OAc, CO ₂ CH ₃ ) ¹ H-NMR (CDCl ₃ , δppm) : 1.20 ~ 1.40 (8H, broad, s), 1.40 ~ 1.70 (4H, broad, m), 1.99 (3H, s, OAc), 2.00 (3H, s, OAc), 2.03 (3H, s, OAc), 2.04 (3H, s, OAc), 2.05 (3H, s, OAc), 2.12 (3H, s, OAc), 2.13 (3H, s, OAc), 2.31 (2H, t, CH ₂ C O ₂ CH ₃ ) , 3.36 ~ 3.90 (6H, m), 3.66 (3H, s, CO ₂ CH ₃ ) , 4.00 ~ 4.36 (3H, m), 4.47 (1H, d, J = 8.3Hz, H-1), 4.84 ~ 5.38 (5H, m), 5.45 (1H, d, J = 2.0Hz, H-1 ′)

Compound (92) (α) ²⁰ _D +122.3 ° (C 1.46, CHCl ₃ ) IR (CHCl ₃ ) cm ^-1 : 1750 (OAc, CO ₂ CH ₃ ) ¹ H-NMR (CDCl ₃ , δppm) : 1.20 ~ 1.40 (8H, broad, s), 1.46 ~ 1.70 (4H, broad, m), 1.98 (3H, s, OAc), 2.05 (9H, s, OAc x 3), 2.08 (3H, s, OAc ), 2.10 (3H, s, OAc), 2.14 (3H, s, OAc), 2.31 (2H, t, CH ₂ C O ₂ CH ₃ ), 3.36 ~ 3.80 (6H, m), 3.67 (3H, s, CO ₂ CH ₃ ) , 3.80 to 4.00 (1H, m), 4.00 to 4.40 (3H, m), 4.87 (1H, d, J = 3.9Hz, H-1), 4.88 to 5.36 (4H, m), 5.45 (1H, d, J = 2.9Hz, H-1 ′)

Example 26 8-Methoxycarbonyloctyl 6-O- (α-D-
Galactopyranosyl) -β-D-glucopyranoside (9
1) Compound (90) 2.0 according to the method described in Example 24.
g (2.48 mmol) in a solution of 20 ml of methanol,
28% sodium methoxide-methanol solution 0.05
The mixture was added with ml and deacetylated to obtain 1.23 g (yield 96.9%) of a colorless caramel target product (91).

[Α] ²⁰ _D + 41.3 ° (C 0.6, H ₂ O) IR (KBr) cm ⁻¹ : 3400 (OH), 1740 (CO ₂ CH ₃ ) ¹ H-NMR (D ₂ O, δppm) : 1.20~1.40 (8H, broad, s ), 1.44~1.68 (4H, broad, m), 2.38 (2H, t, J = 7.3Hz, CH 2 C O 2 CH 3), 3.16~3.28 (1H, m ), 3.30 ~ 3.52 (2H, m), 3.52 ~ 4.00 (11H, m), 3.680 (3H, s, CO ₂ CH ₃ ) , 4.47 (1H, d, J = 8.3Hz, H-1), 4.97 ( 1H, d, J = 2.9Hz, H-1 ′)

Example 27 8-Methoxycarbonyloctyl 6-O- (α-D-
Galactopyranosyl) -α-D-glucopyranoside (9
3) Compound (92) 940 was prepared according to the method described in Example 24.
To a solution of mg (1.17 mmol) in 10 ml of methanol,
28% sodium methoxide-methanol solution 0.05
The mixture was added with ml and deacetylated to obtain 546 mg (yield 91.5%) of the desired product (93) as a colorless caramel.

[Α] ²⁰ _D +114.9 ° (C 0.70, H ₂ O) IR (KBr) cm ⁻¹ : 3400 (OH), 1740 (CO ₂ CH ₃ ) ¹ H-NMR (D ₂ O, δppm) : 1.20~1.40 (8H, broad, s ), 1.44~1.72 (4H, broad, m), 2.38 (2H, t, J = 7.3Hz, CH 2 C O 2 CH 3), 3.16~3.28 (1H, m ), 3.28 ~ 4.00 (13H, m), 3.68 (3H, s, CO ₂ CH ₃ ) , 4.91 (1H, d, J = 3.4Hz, H-1), 4.96 (1H, d, J = 2.9Hz, H-1 ′)

Reference Example 46 8-methoxycarbonyloctyl 2,3,4-tri-
O-acetyl-α-D-xylopyranoside (95), 8
-Methoxycarbonyloctyl 2,3,4-tri-O
-Acetyl-β-D-xylopyranoside (96)

1. J. Am. Chem. Soc., 47, 1280 (1925)
2,3,4-tri-O-acetyl-α-D-xylopyranosyl bromide (94) 1.2 prepared by the method of
To 10 g of 0 g (3.54 mmol) benzene was added 447 mg (1.77 mmol) mercuric cyanide, 1.0 g powdered anhydrous calcium sulfate and 333 mg (1.77 mmol) 8-methoxycarbonyloctanol. The mixture was stirred at room temperature in an argon stream for 24 hours. After filtering the reaction solution, 20 ml of ethyl acetate was added to the filtrate and washed with 20 ml of water. The syrup obtained by dehydrating this with anhydrous magnesium sulfate and concentrating under reduced pressure was purified by silica gel column chromatography (toluene: ethyl acetate = 8: 1).
426 mg of compound (95) (yield 53.9%) and 98 mg of compound (96) (yield 12.4%) were obtained.

2. To a solution of 850 mg (2.51 mmol) of compound (94) in 10 ml of dichloroethane, 1.61 g (6.28 mmol) of silver triflate, 1,1,3,3-
Tetramethylurea 875 mg (7.53 mmol) and 8
-Methoxycarbonyloctanol 236 mg (1.46
Was added and the mixture was stirred at room temperature for 9 hours in an argon stream. The syrup obtained by concentrating the reaction solution under reduced pressure was dissolved in 20 ml of chloroform and washed with 20 ml of water. Syrup obtained by dehydrating this with anhydrous calcium sulfate and concentrating under reduced pressure was purified by silica gel column chromatography (toluene: ethyl acetate = 8: 1) to obtain 375 mg of compound (96).
(Yield 67.0%) was obtained.

Compound (95) [α] ²³ _D + 92.4 ° (C 1.05, CHCl ₃ ) IR (NaCl) cm ⁻¹ : 2940, 1760, 1230 ¹ H-NMR (CDCl ₃ , δppm): 2.03, 2.06 ( 9H, 2s, OAc), 2.31 (2H, t, J = 7.6Hz, C H ₂ C OOCH ₃ ), 3.67 (3H, s, OCH ₃ )

Elemental analysis C ₂₁ H ₃₄ O ₁₀ Calculated value C 56.49% H 7.68% Actual value C 55.98% H 7.97%

Compound (96) (α) ²² _D -19.5 ° (C 0.87, CHCl ₃ ) IR (NaCl) cm ^-1 : 2930, 1760, 1220 ¹ H-NMR (CDCl ₃ , δppm): 2.03, 2.05, 2.06 (9H, 3s, OAc), 2.30 (2H, t, J = 7.4Hz, C H ₂ C OOCH ₃ ), 3.67 (3H, s, OCH ₃ ), 4.46 (1H, d, J = 6.8Hz, H -1)

Elemental analysis C ₂₁ H ₃₄ O ₁₀ Calculated value C 56.49% H 7.68% Actual value C 56.12% H 7.91%

Example 28 8-Methoxycarbonyloctyl α-D-xylopyranoside (97) 410 mg (0.918 mmol) of compound (95) in 4 ml of methanol was added with 28% sodium methylate 0.12.
ml was added, and the mixture was stirred at room temperature for 3 hours. The reaction solution was neutralized with Amberlite IR-120B (H ⁺ type), the precipitate was collected by filtration, and the filtrate was concentrated under reduced pressure. The obtained syrup was purified by silica gel column chromatography (chloroform: methanol = 30: 1) to obtain 243 mg of compound (97) (yield 82.8%).

[Α] ²⁵ _D + 90.9 ° (C 1.10, CH ₃ OH) IR (NaCl) cm ⁻¹ : 3450, 2960, 1740, 1040 ¹ H-NMR (CD ₃ OD, δppm): 2.30 (2H, t, J = 7.6Hz, C H ₂ C OOCH ₃ ), 3.64 (3H, s, OCH ₃ ) 4.69 (1H, d, J = 3.9Hz, H-1)

Elemental analysis C ₁₅ H ₂₈ O ₇ Calculated value C 56.23% H 8.81% Actual value C 56.11% H 9.21%

Example 29 8-Methoxycarbonyloctyl β-D-xylopyranoside (98) 28% sodium methylate 0.03 in a solution of 101 mg (0.226 mmol) of compound (96) in 2 ml of methanol.
ml was added, and the mixture was stirred at room temperature for 3 hours. The reaction solution was neutralized with Amberlite IR-120B (H ⁺ type), the precipitate was collected by filtration, and the filtrate was concentrated under reduced pressure. The obtained syrup was purified by silica gel column chromatography (chloroform: methanol = 30: 1) to give compound (98) 58.3 mg.
(Yield 80.5%) was obtained.

[Α] ²⁴ _D -10.3 ° (C 1.02, CH ₃ OH) IR (NaCl) cm ^-1 : 3450, 2960, 1740, 1290, 1050 ¹ H-NMR (CD ₃ OD, δppm): 2.31 ( 2H, t, J = 7.3Hz, C H ₂ C OOCH ₃ ), 3.65 (3H, s, OCH ₃ ) 4.17 (1H, d, J = 7.6Hz, H-1)

Elemental analysis C ₁₅ H ₂₈ O ₇ Calculated value C 56.23% H 8.81% Actual value C 56.15% H 9.34%

Reference Example 47 3,4-di-O-acetyl-L-fucal (100) Carbohydr. Res., 4, According to the method of 189 (1967), 1,
New 2,3,4-tri-O-acetyl-α-L-fucopyranosyl bromide (99) prepared from 630 mg (1.90 mmol) of 2,3,4-tetra-O-acetyl-L-fucopyranose After cooling 4.2 ml of a 50% aqueous solution of acetic acid of 10 to -10 ° C, 1.26 g of zinc dust and 0.13 g of copper sulfate were added, and the mixture was vigorously stirred at the same temperature for 1 hour. After collecting the precipitate by filtration, the filtrate was extracted twice with 10 ml of chloroform. The extract was washed successively with 20 ml of saturated sodium hydrogen carbonate and 20 ml of water. The syrup obtained by dehydrating this with anhydrous magnesium sulfate and concentrating under reduced pressure was subjected to silica gel column chromatography (toluene: ethyl acetate = 3).
0: 1) to 284 mg of compound (100) (yield 6
9.7%).

[Α] ²² _D +0.9 ° (C 0.98, CHCl ₃ ) IR (NaCl) cm ⁻¹ : 2990, 1750, 1650, 1230 ¹ H-NMR (CDCl ₃ , δppm): 1.28 (3H, d, J = 6.6Hz, H-6), 2.02, 2.17 (6H, 2s, OAc), 4.64 ~ 4.66 (1H, m, H-2), 6.47 (1H, d, J = 8.3Hz, H-1)

Reference Example 48 8-Methoxycarbonyloctyl 3,4-di-O-acetyl-2-azido-2-deoxy-α-L-fucopyranoside (102), 8-methoxycarbonyloctyl
3,4-di-O-acetyl-2-azido-2-deoxy-β-L-fucopyranoside (103)

To a solution of 1.17 g (5.46 mmol) of compound (100) in 30 ml of acetonitrile was added 532 mg (8.19 mmol) of sodium azide and 10.8 g (19.7 mmol) of ceric ammonium nitrate,
The mixture was stirred at -15 ° C for 4 hours. 50 ml of cold water was added to the reaction solution, and the mixture was extracted twice with 30 ml of chloroform. The extract was washed with 50 ml of cold water, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The resulting syrup was purified by silica gel column chromatography (toluene: ethyl acetate = 20: 1) to obtain 982 mg of azide mixture (101) ( Yield 56.6
%) Was obtained. To a solution of 830 mg (2.61 mmol) of mixture (101) in 80 ml of acetonitrile was added lithium bromide 9
07 mg (10.4 mmol) was added, and the mixture was stirred at room temperature for 2 hours. The syrup obtained by concentrating the reaction solution under reduced pressure was suspended in 30 ml of cold water and extracted with 30 ml of chloroform. 331 mg of mercuric cyanide was added to 8 ml of a benzene solution of syrup obtained by dehydration over anhydrous magnesium sulfate and concentration under reduced pressure.
31 mmol), 800 mg anhydrous calcium sulfate and 8-
Methoxycarbonyloctanol (246 mg, 1.31 mmol) was added, and the mixture was stirred at room temperature for 17 hours in an argon stream. After filtering the reaction solution, 20 ml of ethyl acetate was added to the filtrate and washed with 30 ml of water. The syrup obtained by dehydrating this with anhydrous magnesium sulfate and concentrating under reduced pressure was purified by silica gel column chromatography (n-hexane: ethyl acetate = 6: 1) to obtain 187 mg of compound (102) (yield 32.3%). ) And 249 mg of compound (103) (yield 4
3.0%) was obtained.

Compound (102) [α] ²³ _D -121.8 ° (C 1.02, CHCl ₃ ) IR (NaCl) cm ⁻¹ : 2930, 1760, 1220 ¹ H-NMR (CDCl ₃ , δppm): 1.14 (3H, d, J = 6.6Hz, H-6), 2.06, 2.17 (6H, 2s, OAc), 2.31 (2H, t, J = 7.3Hz, C H ₂ C OOCH ₃ ), 3.58 (1H, dd, J = 3.4, 11.2Hz, H-2), 3.67 (3H, s, OCH ₃ ), 4.94 (1H, d, J = 3.4Hz, H-1), 5.30 (1H, m, H-4), 5.37 (1H , dd, J = 3.4, 11.2Hz, H-3)

Compound (103) [α] ²² _D -3.1 ° (C 0.91, CHCl ₃ ) IR (NaCl) cm ⁻¹ : 2930, 1760, 1220 ¹ H-NMR (CDCl ₃ , δppm): 1.21 (3H, d, J = 6.4Hz, H-6), 2.05, 2.17 (6H, 2s, OAc), 2.30 (2H, t, J = 7.3Hz, C H ₂ C OOCH ₃ ), 3.64 (1H, dd, J = 8.1, 11.0Hz, H-2), 3.67 (3H, s, OCH ₃ ), 4.32 (1H, d, J = 8.1Hz, H-1), 4.77 (1H, dd, J = 3.4, 11.0Hz, H -3) 5.18 (1H, m, H-4)

Example 30 8-Methoxycarbonyloctyl 2-acetamido-
2-Deoxy-α-L-fucopyranoside (104) 180 mg (0.406 mmol) of compound (102) in 10 ml of ethanol was added with sodium borohydride 46.1.
mg (1.22 mmol) and nickel chloride (hexahydrate)
29 mg was added, and the mixture was stirred at room temperature for 4 hours. After filtering the reaction solution, the syrup obtained by concentrating the filtrate under reduced pressure was purified by silica gel column chromatography (chloroform: methanol: concentrated aqueous ammonia = 50: 10: 1). 0.5 ml of acetic anhydride was added to a 1 ml solution of the obtained compound in methanol, and the mixture was stirred overnight at room temperature. The syrup obtained by concentrating the reaction solution under reduced pressure was purified by silica gel column chromatography (chloroform: methanol = 20: 1) to give the compound (10
4) 111 mg (yield 73.0%) was obtained.

[Α] ²⁰ _D -158.3 ° (C 0.53, CH ₃ OH) mp 92 ° C IR (KBr) cm ^-1 : 3300, 2930, 1740, 1650, 1050 ¹ H-NMR (CD ₃ OD, δppm): 1.21 (3H, d, J = 6.6Hz, H-6), 1.97 (3H, s, NAc), 2.31 (2H, t, J = 7.3Hz, C H ₂ C OOCH ₃ ), 3.64 (3H, s, OCH ₃ ), 3.75 (1H, dd, J = 3.4, 11.1Hz, H-3), 4.19 (1H, dd, J = 3.9, 11.1Hz, H-2), 4.73 (1H, d, J = 3.9Hz , H-1)

Example 31 8-Methoxycarbonyloctyl 2-acetamido-
2-Deoxy-β-L-fucopyranoside (105) 240 mg (0.541 mmol) of compound (103) was treated in the same manner as in Example 30 to obtain 141 mg (6) of compound (105).
9.9%).

[Α] ²⁰ _D +3.9 ° (C 0.65, CH ₃ OH) IR (KBr) cm ⁻¹ : 3300, 2930, 1730, 1650, 1070 ¹ H-NMR (CD ₃ OD, δppm): 1.26 ( 3H, d, J = 6.6Hz, H-6), 1.96 (3H, s, NAc), 2.30 (2H, t, J = 7.3Hz, C H ₂ C OOCH ₃ ), 3.64 (3H, s, OCH ₃ ), 4.31 (1H, d, J = 8.3Hz, H-1)

Reference Example 49 3-Benzyloxycarbonylpropyl 2,3,3
4,6-Tetra-O-benzyl-β-D-mannopyranoside (110) 2,3,4,6-tetra-O-benzyl-α-D-mannopyranosyl chloride (106) 580 mg (1.04) 3 ml of anhydrous methylene chloride in 3 ml of benzyl 4-hydroxybutanoic acid benzyl ester (107) (544 mg, 2.
8 mmol), silver carbonate 500 mg, calcium sulfate 500
A mixture of 10 mg of anhydrous methylene chloride and 10 ml of anhydrous methylene chloride was added dropwise under a nitrogen stream, and the mixture was reacted at room temperature for 8 hours. After completion of the reaction, the insoluble matter was removed by filtration, and the filtrate was distilled off under reduced pressure to obtain a crude product 1.11.
4 g was obtained. This was separated and purified by flash chromatography (hexane: ethyl acetate = 6: 1) to obtain 462 mg of compound (110) (yield 63.8%).

[Α] ²⁵ _D -41.68 ° (C 0.95, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 3060, 3040, 1740

Reference Example 50 11-Methoxycarbonylundecyl 2,3,4,6
-Tetra-O-benzyl-β-D-mannopyranoside (111) Chloride (106) is condensed with alcohol (108) in the same manner as in the synthesis of compound (110) to give compound (111) 489. 3 mg (yield 62.5%) was obtained.

[Α] ²⁵ _D 41.75 ° (C 1.20, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 1740

Reference Example 51 14-methoxycarbonyltetradecyl 2,3,4
6-Tetra-O-benzyl-β-D-mannopyranoside (112) Chloride (106) and alcohol (109) were condensed to 478.2 mg of compound (112) in exactly the same manner as the synthesis of compound (110). (Yield 57.8%) was obtained.

[Α] ²⁵ _D −38.90 ° (C 1.09, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 1740

Example 32 3-Methoxycarbonylpropyl-β-D-mannopyranoside (114) Compound (110) 758 mg (1.05 mmol), 10
% A mixture of 530 mg of palladium-carbon, 10 ml of methanol and 10 ml of ethyl acetate was shaken under a hydrogen stream at about 5 atm for 7 hours. After completion of the reaction, the catalyst was filtered off and the filtrate was distilled off under reduced pressure to obtain Compound (113) in 20 ml of methanol.
Dissolve in, add excess diazomethane / ether solution,
Stir for 30 minutes. After decomposing excess diazomethane with a small amount of acetic acid, the solution was distilled off under reduced pressure to obtain a crude product. Isopropyl ether was added to this to crystallize the compound (11
238 mg of 4) was obtained as colorless crystals.

Mp 102 to 108 ° C., [α] ¹⁴ _D −46.15 ° (C 1.04, CH ₃ OH) IR (KBr) cm ⁻¹ : 3450, 3300, 1730 ¹ H-NMR (CD ₃ OD, δppm): 1.85 ~ 1.95 (2H, m), 2.45 (2H, t, J = 7.5Hz), 3.16 ~ 3.21 (1H, m), 3.41 ~ 3.45 (1H, dd), 3.48 ~ 3.58 (2H, m), 3.65 ( 3H, s, CO ₂ CH ₃ ), 3.67 ~ 3.72 (1H, dd), 3.83 (1H, d, J = 3.2Hz), 3.84 ~ 3.88 (1H, dd), 3.88 ~ 3.95 (1H, m), 4.48 (1H, d, J = 0.7Hz)

Example 33 11-Methoxycarbonylundecyl-β-D-mannopyranoside (115) Compound (111) 631 mg (0.84 mmol), 10
% Palladium-carbon (500 mg) and methanol (20 ml) were shaken in a hydrogen stream at about 5 atm to react overnight. After completion of the reaction, the catalyst was removed by filtration and the filtrate was distilled off under reduced pressure to obtain 196.3 mg of compound (115) (yield 59.5).
%)Obtained.

Mp 124.5 to 125 ° C., [α] ¹⁸ _D −32.63 ° (C 1.14, CH ₃ OH) IR (KBr) cm ⁻¹ : 3400, 3300, 1735 ¹ H-NMR (CD ₃ OD, δppm): 1.26 ~ 1.40 (14H, broad s), 1.50 ~ 1.65 (4H, m), 2.30 (2H, t, J = 7.5Hz), 3.16 ~ 3.21 (1H, m), 3.41 ~ 3.45 (1H, dd), 3.48 ~ 3.58 (2H, m), 3.64 (3H, s, CO ₂ CH ₃ ), 3.67 ~ 3.72 (1H, dd), 3.83 (1H, d, J = 2.9Hz), 3.84 ~ 3.88 (1H, dd), 3.88 ~ 3.92 (1H, m), 4.48 (1H, d, J = 0.74Hz, H-1)

Example 34 14-Methoxycarbonyltetradecyl-β-D-mannopyranoside (116)

A mixture of 620 mg (0.78 mmol) of compound (112), 500 mg of 10% palladium-carbon and 20 ml of methanol was shaken overnight in a hydrogen stream at about 5 atm. After the reaction was completed, the catalyst was removed by filtration, and the filtrate was evaporated under reduced pressure to give 170.7 mg of compound (116) (yield: 50.4
%)Obtained.

[Α] ¹⁸ _D -26.22 ° (C 0.98, CH ₃ OH) IR (CHCl ₃ ) cm ⁻¹ : 3400, 1730 ¹ H-NMR (CD ₃ OD, δppm): 1.26 to 1.40 (20H, broad s), 1.50 ~ 1.65 (4H, m), 2.30 (2H, t, J = 7.5Hz), 3.16 ~ 3.21 (1H, m), 3.41 ~ 3.45 (1H, dd), 3.48 ~ 3.58 (2H, m) , 3.64 (3H, s, CO ₂ CH ₃ ), 3.67 ~ 3.72 (1H, dd), 3.83 (1H, d, J = 2.9Hz), 3.84 ~ 3.88 (1H, dd), 3.88 ~ 3.92 (1H, m ), 4.48 (1H, d, J = 0.7Hz)

Reference Example 52 Benzyl 2-acetamido-3-O- (2,3,4)
6-Tetra-O-acetyl-β-D-galactopyranosyl) -2-deoxy-α-D-glucopyranoside (11
9)

2.26 g (6.43 mmol) of benzyl 2-acetamido-2-deoxy-4,6-O-isopropylidene-α-D-glucopyranoside (118),
Mercury cyanide 1.67 g, anhydrous nitromethane 100
a mixture of 100 ml of anhydrous benzene and 130 ml of anhydrous benzene is heated to reflux,
About 30 ml of benzene was distilled off. Next, the reaction solution is 60 to 65
Acetobromogalactose (117) 3 while maintaining at ℃
A 10 ml solution of g of benzene was slowly added dropwise. 6 after dropping
After stirring for an hour, 250 mg of mercuric cyanide and 550 mg of acetobromogalactose / 10 ml of benzene were added, and the mixture was stirred for 20 hours. After the reaction was completed, the insoluble matter was removed by filtration, water was added to the filtrate, and the mixture was shaken well to separate the organic layer.
After washing with saturated saline, it was dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure to obtain a crude product. 2 to this
Crystallization was carried out by adding propanol-isopropyl ether, and the compound (119) was obtained in an almost pure state in an amount of 3.375 g (81.8%).

Mp 193-195 ° C., (α) ²⁵ _D + 73.5 ° (C 1.0, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 3500, 3450, 1750, 1690 ¹ H-NMR (CDCl ₃ , δppm) : 1.95, 1.97, 2.05, 2.07, 2.15 (3H, s, Ac), 3.54 ~ 3.65, 3.68 ~ 3.73 (each 2H, each m), 3.73 ~ 3.80, 3.84 ~ 3.90, 3.93 ~ 3.98 (each 1H, each m ), 4.05 to 4.18 (2H, m), 4.28 (1H, dt), 4.52 (1H, d, J = 8.1Hz), 4.45, 4.73 (2H, J _AB = 11.5Hz, OCH ₂ Ph), 4.98, 5.16 ~ 5.23, 5.37 (each 1H, each dd), 5.52 (1H, d, J = 10.0Hz, NH), 7.30 ~ 7.45 (5H, m)

Elemental analysis C ₂₉ H ₃₉ NO ₁₅ Calculated value C 54.29% H 6.13% N 2.18% Actual value C 54.01% H 6.06% N 2.01%

Reference Example 53 Benzyl 2-acetamido-3,6-di-O- (2,
3,4,6-Tetra-O-acetyl-β-D-galactopyranosyl) -2-deoxy-α-D-glucopyranoside (121)

3.2 g (5 mmol) of compound (119), 1.5 g of mercuric cyanide, 80 ml of anhydrous benzene.
A mixture of 60 ml of anhydrous nitromethane and 60 ml of anhydrous nitromethane was heated under reflux with stirring to distill off about 20 ml of benzene. Then under a nitrogen stream,
Acetobrom galactose 2.1 at an internal temperature of 60-65 ° C
A solution of 10 g of benzene (5 mmol) in benzene was added dropwise and the reaction was carried out with stirring. After 15 hours, 260 mg of mercuric cyanide and 410 mg of acetobromogalactose / 5 ml of benzene were added again, and the mixture was further stirred for 2 hours. After completion of the reaction, insoluble matter was removed by filtration through Celite, and the solvent was distilled off under reduced pressure to obtain 5.87 g of a crude product. This was separated and purified by flash chromatography (ethyl acetate: hexane = 3: 1) to give 1.647 g of compound (120), 598 mg of a mixture of compounds (120) and (121), and 1 of compound (121). 0.553 g was obtained. The mixture was further separated and purified by flash chromatography (ethyl acetate: hexane = 3: 1), and combined with the one obtained above, 1.647 g of compound (120) (yield 45.4).
%) And 1.876 g of the compound (121) (yield 38.6).
%) Was obtained.

Compound (121) [α] ²⁶ _D + 39.9 ° (C 1.0, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 3500, 3450, 1750, 1685 ¹ H-NMR (CDCl ₃ , δppm): 1.94 , 1.96, 1.97, 2.00, 2.03, 2.05, 2.05, 2.12, 2.14 (each 3H, each s, Ac), 3.42 (1H, t, J = 9.0Hz), 3.55 (1H, s, OH), 3.56 to 3.60 (1H, m), 3.62 to 3.68 (1H, dd), 3.79 to 3.86 (1H, m), 3.89 to 3.98 (2H, m), 4.05 to 4.20 (4H, m), 4.21 to 4.29 (2H, m) , 4.38, 4.70 (2H, J _AB = 11,5Hz, OCH ₂ Ph), 4.48 (1H, d, J = 8.1Hz), 4.57 (1H, d, J = 8.1Hz), 4.79 (1H, d, J = 3.7Hz, H-1), 4.95, 5.02 (each 1H, dd, J = 3.4Hz, 10.5Hz), 5.15 ~ 5.21 (1H, dd), 5.22 ~ 5.28 (1H, dd), 5.33 ~ 5.36, 5.37 ~ 5.40 (1H, m), 5.50 (1H, d, J = 9.8Hz, NH), 7.25 ~ 7.40 (5H, m)

Reference Example 54 2-acetamido-1,4-di-O-acetyl-3,6
-Di-O- (2,3,4-6-tetra-O-acetyl-
β-D-galactopyranosyl) -2-deoxy-D-glucopyranose (122)

1.47 g (1.51 mmol) of compound (121), 30 ml of acetic acid and 54% of 10% palladium-carbon.
0 mg was reacted for 19 hours under hydrogen stream. After the reaction,
The catalyst was removed by filtration, and the filtrate was evaporated under reduced pressure. To the crude product obtained, 10 ml of pyridine and 2 ml of acetic anhydride were added, and the mixture was stirred at room temperature for 7 minutes.
Stir for 2 hours. After completion of the reaction, the solvent was distilled off under reduced pressure to obtain 1.766 g of a crude product. This was purified by flash chromatography (ethyl acetate) to give the compound (12
2.337 g (yield 91.6%) of 2) was obtained.

[Α] ²⁶ _D + 14.9 ° (C 1.0, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 3450, 1750, 1690 ¹ H-NMR (CDCl ₃ , δppm): 1.95, 1.96, 1.99, 2.03 , 2.04 (each 3H, s, Ac), 2.04 (6H, s, Ac × 2), 2.06, 2.11, 2.13, 2.18 (each 3H, s, Ac), 3.42 ~ 3.48 (1H, dd), 3.84 ~ 3.96 (4H, m), 3.98 ~ 4.04, 4.06 ~ 4.12, 4.12 ~ 4.18, 4.18 ~ 4.25 (each 1H, m), 4.45 (1H, d, J = 8.1Hz), 4.45 ~ 4.52 (1H, m), 4.57 (1H, d, J = 7.6Hz), 4.78 ~ 4.86 (1H, m), 4.90 ~ 5.06 (4H, m), 5.10 ~ 5.18 (1H, m), 5.32 ~ 5.38 (3H, m), 6.03 (1H , d, J = 3.7Hz)

Reference Example 55 8-Methoxycarbonyloctyl 2-acetamido-
3,6-di-O- (2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl) -2-deoxy-
β-D-glucopyranoside (123)

966 mg (1 mmol) of compound (122)
20 ml of acetyl chloride solution was cooled to -10 ° C, dry hydrogen chloride gas was passed through for 1 hour, and then left at room temperature overnight. After completion of the reaction, acetyl chloride was distilled off under reduced pressure, and anhydrous benzene was added to dissolve the crude product. After the solvent was completely distilled off under reduced pressure, the obtained crude chloride was dissolved in 15 ml of methylene chloride, followed by stannous trifluoromethanesulfonate (4).
20 mg and 1.5 g of molecular sieve 4A were added, and the mixture was stirred under a nitrogen stream for 30 minutes. 400 mg of 8-methoxycarbonyloctanol and 1 part of tetramethylurea were added to this mixture.
A solution of 90 mg and 5 ml of methylene chloride was added dropwise and stirred for 17 hours. After completion of the reaction, 10 ml of chloroform was added, the mixture was filtered through Celite, the filtrate was washed with water, and the chloroform layer was separated.
It was dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure to obtain 1.15 g of a crude product. This was purified by flash chromatography (ethyl acetate) to give compound (1
23) was obtained (244 mg, yield 22.3%).

[Α] ²⁴ _D -5.0 ° (C 1.0, CHCl ₃ ) IR (CHCl ₃ ) cm ⁻¹ : 3460, 1750, 1680 ¹ H-NMR (CDCl ₃ , δppm): 1.97, 1.98, 2.01, 2.05 , 2.05, 2.06, 2.08, 2.15, 2.15 (each 3H, s, Ac), 1.30 (10H, broad, s), 1.50 ~ 1.70 (4H, m), 2.31 (2H, t, J = 7.6Hz), 3.10 ~ 3.20, 3.40 ~ 3.50, 3.50 ~ 3.60 (1H, m), 3.67 (3H, s, CO ₂ CH ₃ ), 3.80 ~ 3.95 (3H, m), 4.00 ~ 4.20, 4.45 ~ 4.55 (each 1H, m) , 4.55 ~ 4.60 (2H, m), 4.70 ~ 4.80, 4.80 ~ 4.90 (each 1H, m), 4.90 ~ 5.04 (2H, m), 5.04 ~ 5.10, 5.15 ~ 5.25 (each 1H, m), 5.72 (1H , d, J = 6.35Hz)

Example 35 8-Methoxycarbonyloctyl 2-acetamido-
3,6-di-O- (β-D-galactopyranosyl) -2
-Deoxy-β-D-glucopyranoside (124) 28% sodium methoxide-methanol 0.05 ml in a solution of 224.4 mg of compound (123) 224.4 mg in methanol.
Was added, and the mixture was stirred at room temperature for 24 hours. After completion of the reaction, 5 ml of Amberlite IR120 (H ⁺ type) -methanol was added, and the mixture was stirred for 30 minutes for neutralization and then filtered to remove the resin. Then, the filtrate was distilled off under reduced pressure to obtain 138 mg (yield 94.0%) of Compound (124) as a colorless powder.

Mp 233-234 ° C. (CH ₃ OH), [α] ²⁵ _D -18.9 ° (C 1.0, CH ₃ OH) IR (KBr) cm ⁻¹ : 3350, 1730, 1650 ¹ H-NMR (CD ₃ OD, δppm): 1.31 (10H, broad s), 1.48 ~ 1.65 (4H, m), 1.95 (3H, s, NHAc), 2.30 (2H, t, J = 7.6Hz), 3.42 ~ 3.58 (10H, m ), 3.64 (3H, s, CO ₂ CH ₃ ), 3.65 ~ 3.90 (H, m), 4.16 (1H, dd), 4.26, 4.34 (1H, d, J = 7.6Hz), 4.46 (1H, d, (J = 8.3Hz)

[0330] Elemental analysis _{C 30 H 53 NO 18 · 2H} 2 O Calculated C 47.93% H 7.64% N 1.86 % Found C 47.40% H 7.38% N 1.70 %

Example 50 N-Acetylmannosaminyl-β-oxyoctanoyl albumin (200)

To a solution of 85 mg (0.217 mmol) of compound (21) in 1 ml of ethanol was added hydrazine hydrate 0.2.
ml was added and left at room temperature for 24 hours. The reaction mixture was concentrated to dryness under reduced pressure. A small amount of ethanol and water were added to the residue to dissolve it, and the residue was concentrated to dryness again under reduced pressure to remove excess hydrazine hydrate. The obtained acid hydrazide 8
5 mg was dissolved in 2.9 ml dimethylformamide and cooled to -25 ° C under a nitrogen stream. After adding 0.19 ml of 4.1N hydrochloric acid-dioxane solution, 28 mg of tert-butyl nitrite
0.28 ml of the dimethylformamide solution of was added and stirred for 30 minutes. 0.3 ml of a dimethylformamide solution containing 20 mg of sulfamic acid was added, and the mixture was stirred for 5 minutes. Then, the solution containing this acid azide was added dropwise to 27 ml of 0.08M sodium borate-0.35M potassium hydrogen carbonate buffer solution containing 269 mg of human serum albumin under ice cooling while slowly stirring. The reaction mixture was allowed to stand at 4 ° C. for 24 hours, after which Spectrapol (molecular weight cutoff 12,000-1
4,000) dialysis membrane at 2 ℃ against purified water 2
It was dialyzed for 4 hours. Take out the dialysis solution, freeze to dryness, and
270 mg of -acetylmannosaminyl-β-oxyoctanoyl albumin was obtained. N-acetylmannosaminyl-β-oxyoctanoyl albumin was hydrolyzed with 4N hydrochloric acid and then titrated by the Elson-Morgan method to give 4
It was revealed that three N-acetylmannosaminyloctyl groups were introduced.

Example 51 Compounds (9), (11), (12), (20) and (2
2), (26), (27), (34), (35), (3
6), (37), (41), (48), (53), (5
8), (62), (66), (68), (70), (7
3), (77), (88), (89), (91), (9
3), (97), (98), (104), (105),
Using (114), (115), (116) and (124), a sugar residue was introduced into human serum albumin by the same method as described in Example 50. The introduced sugar content was measured by the Elsson-Morgan method after hydrolysis with a phenol-sulfuric acid method in the case of neutral sugar, and with 4N hydrochloric acid in the case of hexosamine. The compounds and sugar-modified albumin sugar contents are listed in Tables 1 and 2.

[0334]

[Table 1]

[0335]

[Table 2]

The physical constants of these compounds are shown in Tables 3 and 4.

[0337]

[Table 3]

[0338]

[Table 4]

The measurement of the sugar content and the liquid chromatography test method are as follows.

Sugar content of neoglycoprotein (200)
Measurements human serum albumin per mole of 2-acetamido-2
Deoxy-D-mannose content was quantified by the Elson-Morgan method. 2.48 mg of compound (200) was dissolved in 0.25 ml of 4N hydrochloric acid and heated at 100 ° C. for 4 hours. The reaction solution was concentrated to dryness under reduced pressure. The residue obtained is purified water 2
Dissolve in 0.5 ml, transfer 0.5 ml to a test tube with a glass stopper, add 1.0 ml of acetylacetone reagent, and add 1 at 90 ° C.
Heated for hours. After cooling with water, 10 ml of 96% ethanol was gently added, and then 1.0 ml of Ehrlich reagent was added and stirred. After mixing, the mixture was allowed to stand at room temperature for 1 hour and colorimetrically determined at 530 nm. From the calibration curve obtained by similarly treating 2-acetamido-2-deoxy-D-mannose, compound (200)
The 2-acetamido-2-deoxy-D-mannose content of was determined to be 43.

Sugar content of neoglycoprotein (217)
The number of moles of fucose bound to 1 mole of human serum albumin in the measurement compound (217) was determined by the phenol-sulfuric acid method (Michel Dubois et al., Anal. Chem., 28,
350-356 (1956)).

2.1 mg of compound (217) was dissolved in distilled water, 1 ml of 5% phenol aqueous solution was added, and then 5 ml of concentrated sulfuric acid.
Was quickly added to the solution. This was left at room temperature for 10 minutes and then kept at 25 ° C to 30 ° C for 20 minutes. The absorbance (wavelength: 480 nm) of this solution was measured, and the sugar content was determined from the standard curve of fucose prepared according to the method in the literature.
As a result, the binding mole number of fucose to 1 mole of human serum albumin was determined to be 22.

Liquid Chromatography Test Method The molecular weight of compound (200) was measured by gel filtration liquid chromatography. The following equipment and columns were used. Pump: JASCO 880-PV, Detector: JASCO 870-UV, Column: TSK-GEL G3000SW.

As analytical conditions, eluent: 0.008M isotonic phosphate buffer pH 7.4, flow rate: 0.4 ml / min, measurement wavelength: 280 nm was used.

Compound (200) was dissolved in phosphate buffer so that the concentration was 1 mg / ml, and the solution was added to the 20 μl column.

As a result, it was shown that the compound (200) was eluted at 20.6 minutes. In addition, the elution times of proteins having different molecular weights, β-amylase (molecular weight 200,000), human serum albumin (molecular weight 67,000) and carbonic anhydrase (molecular weight 29,000) were measured under the same conditions. 19.5, 2
It was 1.8 and 26.7 minutes, and it was shown that the elution time was different depending on the molecular weight. Therefore, Compound (120) was shown to be a compound having a slightly higher molecular weight than human serum albumin.

The elution times of the compounds (202) to (225) were measured under the same conditions. The results are also shown in Tables 3 and 4. As a result, compounds (202) to (22
It was found that 5) had almost the same molecular weight.

Reference Example 56 N-Acetylmannosaminyloxyoctyl Human Serum Albumin Complex with Methotrexate

Biol. Cell., 51, 187 (1984), a dimethylformamide solution of the active ester of N-hydroxysuccinimide of methotrexate synthesized by the method described in
N-acetylmannosaminyl-β-oxyoctyl human serum albumin (200) was added to a phosphate buffer under ice cooling,
The mixture was stirred at the same temperature for 4 hours. The reaction solution was centrifuged, insoluble matter was filtered off, and then filtered using a Millipore filter.
The filtration was separated and purified by gel filtration (G-100) column chromatography to obtain a complex of methotrexate and N-acetylmannosaminyl-oxyoctyl human serum albumin.

Test Example The neoglycoproteins obtained in Examples 50 and 51 were iodinated using the chloramine T method. as a result,
Iodolabeled neoglycoprotein with a specific activity of 18-30 MBq / 20 μg protein was obtained. TCA treatment was performed to confirm that radioactive iodine was bound to the protein. That is, after appropriately diluting with 0.1 M phosphate buffer containing 0.5% bovine serum albumin, 300 μl thereof
Was taken in a test tube, 600 μl of a 15% TCA (trichloroacetic acid) solution was added, and the mixture was stirred and then centrifuged at 3,000 rpm for 10 minutes to measure the activity of radioactive iodine present in the supernatant and the precipitate. As a result, it was found that 95% or more of the total radioactivity was present in the precipitate, and radioactive iodine was bound to the protein.

A distribution experiment in animals was carried out using this iodolabeled neoglycoprotein. First, 4 μg of iodolabeled neoglycoprotein was added to 0.1 M phosphate buffer containing 0.5% bovine serum albumin (pH 7.4).
It was dissolved in 1 ml to give a test solution for administration. When the specific activity of iodine label was high, it was diluted with unlabeled neoglycoprotein and used as a test solution for administration.

This solution was added to SD male rats (body weight: 240).
˜300 g) was administered through the femoral vein to give 0.1 ml per 100 g of body weight. After that, exactly 1 minute, 2 minutes,
Blood is collected from the jugular vein 3 minutes later, and 5 minutes after administration, systemic blood is collected from the inferior aorta and bleeding to death. Immediately thereafter, the heart, lung, thymus, spleen, kidney, muscle, bone marrow, skin and liver were collected.

1 min, 2 min, 3 min, 5 min serum and 5 min whole blood, heart, lung, thymus, spleen, kidney, muscle,
Bone marrow, skin and liver were accurately weighed and their radioactivity was measured.

Further, in order to confirm that the radioactivity present in the administration test solution and the serum 5 minutes after the administration was bound to neoglycoprotein, TCA treatment was carried out. The procedure was performed as described above, and it was confirmed that 90% or more of the total radioactivity was bound to neoglycoprotein.

Next, the obtained serum concentration (1 minute after administration,
From the results of 2 minutes, 3 minutes, and 5 minutes), the area under serum concentration (AUC _0-5 ) from 0 minute to 5 minutes after administration was calculated using the trapezoidal approximation method. AUC _0-5 is defined by the following equation.
AUC _0-5 = ∫ ⁵ ₀ Cdt where C is the serum concentration at neoglycoprotein time (t).

Further, the tissue distribution clearance was calculated from the concentration of neoglycoprotein in each tissue and the ratio of AUC _0-5 . The results are shown in Table 5.

As is clear from the results in Table 5, it was found that modification of albumin with sugar chains increased distribution clearance to bone marrow. In addition, among them, mannose (203), mannosamine (200), fucose (217), mannose α (1-6) mannose (208) and mannose β (1-6) mannose (2
In the neoglycoprotein modified in 09), the distribution clearance to the bone marrow was greatly increased, indicating the directivity to bone marrow tissue.

This indicates that the glycosyl-protein derivative is useful as a drug delivery carrier targeting bone marrow tissue.

[0359]

[Table 5]

Examining the tissue distribution of the above compound,
There are two main groups. Those that disappear from blood immediately after intravenous injection and have the property of being distributed to tissues such as bone marrow and liver, and those that exist in blood for a relatively long time and that slowly distribute to tissues.

In the case of the compound having the above-mentioned characteristic, the larger the distribution clearance to the bone marrow, the quicker and the more the distribution clearance is distributed to the bone marrow. Therefore, it can be evaluated by comparing the distribution clearance to the bone marrow. The compounds belonging to this are (200), (202), (2
04), (208) (209), (217), (21
8), which was about 7 to 11 times higher than that of serum albumin (HSA).

Further, in the case of the compound having the property of (3), if the distribution clearance to the bone marrow is small, but the distribution clearance to other tissues is also small, the compound will be more concentrated in the bone marrow, so that the distribution clearance to the bone marrow will be It can be evaluated by comparing the ratio of distribution clearance to the whole body. Compounds belonging to this group include (204) and (20
5), (212), (213), (215), (21
9), which was about 1.3 to 1.8 times higher than that of HSA.

Example 52 A sugar residue was introduced into human serum albumin by the same method as that described in Example 50 to obtain the compounds 234 to 249 shown in Table 6. The physical constants for each compound are shown in Table 7.

[0364]

[Table 6]

[0365]

[Table 7]

Test Example 2 Compound 226 obtained in Example 51 and compound 2 obtained in Example 52
36, 238, 242, 245 and 246 were prepared as specimen samples, and HSA was prepared as a control sample.

[0367] K, L, Audus et al's method (Pharm.Res., 3,8 1 (19
86)) bovine brain capillary endothelial cells (hereinafter BB)
Abbreviated as CECs) 5 × 10 ⁴ cells / cm ² 6 well
After culturing for 11 days on a (9.6 cm ² ) culture plate, M containing 20 mM HEPES 0.1% phosphate buffer was added.
It was washed 3 times with EM (pH 7.4) (hereinafter abbreviated as reaction medium). After adding 2 ml of reaction medium and preincubating for 30 minutes, ¹²⁵ I-labeled sample ( ¹²⁵ I-ligand) was added at a concentration of 25 ng / ml to 3 μg / ml, especially 20 μg / ml for compounds 242 and 245. At concentrations up to and were incubated. Thereafter, the remaining reaction medium was removed, and the mixture was washed 3 times with a phosphate buffer solution, and 1 ml of 0.1N sodium hydroxide was added. BB solubilized after standing at 4 ° C for 30 minutes
The CECs were collected, and the radioactivity was measured with a gamma counter, and the amount of protein was measured with the Bio-Rad protein assay kit.
Bound).

Separately, an excess amount (100 to 30) is added to each reaction system.
An unlabeled sample (ligand) (0-fold amount) was added, incubated, treated in the same manner as above, and measured to determine the nonspecific uptake activity (+ Cold) per mg of protein. The specific uptake activity (SB; the amount of ligand per mg of incorporated protein) was calculated by subtracting the nonspecific uptake activity from the uptake activity.

The results are shown in FIGS. ● Marked line, ○
The solid lines and solid lines represent uptake activity, nonspecific uptake activity and specific uptake activity, respectively. 1 to 5
Are HSA, compounds 236 and 22 respectively in the order of the numbers in the figure.
6, 238 and 246 show the results of the uptake activity, FIGS. 6 and 8 show the results of the uptake activity of Compound 245, and FIGS. 7 and 9 show the results of the uptake activity of Compound 242.

From FIGS. 1 to 9, compounds 236, 226,
Saturation was observed for 238 and 246 at an addition concentration of 3 μg / ml and for compound 245 at an addition concentration of 10 μg / ml, respectively.
While it has no uptake activity for BCECs,
It was confirmed that all the specimen samples had the uptake activity, and in particular, the uptake activity of the compound 242 did not reach the saturation even at the addition concentration of 20 μg / ml.

Table 8 shows a comparison of the uptake activity at the addition concentration of 3 μg / ml. According to this, it was recognized that there was a difference in uptake recognition for BBCECs depending on the type of sugar. Therefore, it is known that the type of sugar may be appropriately selected according to the purpose and conditions for delivering a predetermined drug to the brain.

[0372]

[Table 8]

Test Example 3 Compounds 236, 237, 240, 241 obtained in Example 52,
243 and 244 were prepared as samples. Sample labeled with ¹²⁵ I in Test Example 2 ( ¹²⁵ I-ligand)
Was added at a concentration of 100 ng / ml, and the uptake activity, nonspecific uptake activity and specific uptake activity were determined in the same manner as in Test Example 2. The result is shown in Figure 1.
0 and FIG. FIG. 10 shows compounds 236 and 23
7, that is, the results obtained for a series of compound groups in which Manβ is a sugar species and only the m number is different, and shows the relationship between the m number and the specific uptake activity. FIG. 11 shows compound 240,
24 shows the results obtained for a series of compounds having 241, 243, and 244, that is, Fucβ as a sugar species and differing only in the m number, and represents the relationship between the m number and the specific uptake activity. From FIG. 10 and FIG. 11, it was confirmed that the activity increased as the m number, that is, the sugar density increased. Therefore, it is known that the sugar density may be appropriately selected according to the purpose and conditions for delivering a predetermined drug to the brain.

Test Example 4 The compound 245 obtained in Example 52 was used as a sample. The compound 226 obtained in Example 51 and the compound 2 obtained in Example 52 were added as unlabeled samples for the purpose of inhibiting the uptake of the specimen sample.
36, 238, 242, 245, 246, 247, 24
8 and 249 and HSA were prepared.

Test Example 2 except that the sample sample labeled with ¹²⁵ I ( ¹²⁵ I-ligand) was added at a concentration of 1 μg / ml, and each unlabeled sample was added uniformly in 100-fold amount. The same procedure as in 2 was performed to determine the uptake activity. Results are shown in FIG. FIG. 12 shows the relationship between the compound added for the purpose of inhibition and the uptake activity of the specimen sample 245. From FIG. 12, it is confirmed that some compounds show an inhibitory effect on the uptake of the specimen sample, but the strongest inhibitory effect is brought about by the specimen sample itself, and the uptake is sugar-specific.

Test Example 5 (Biodistribution Experiment) Compounds 237, 241, 243 and 245 obtained in Example 52
Was prepared as a sample and HSA was prepared as a control sample.
1 mg of each sample was weighed and dissolved in 5 ml of 0.05M borate buffer. On the other hand, to mix the N- Sakushiniimijiru ^[2,3-3 H] DMF solution 100μl treated molecular sieves in toluene solution 100μl propionate was added to the borate buffer. After stirring overnight at 4 ° C, the toluene layer was removed, and the aqueous layer was about 200 µl with Centricon 30.
It was concentrated to. The concentrated solution was subjected to gel filtration using a PD-10 column with a phosphate buffer solution as an eluent to collect radioactivity eluted in the void fraction. The protein concentration of this recovered liquid was measured, and each sample was added to give a concentration of 1 mg / ml, which was used as a liquid for administration to animals.

The H ³ -converted sample was administered to rats (male SD strain 6 weeks) at 1 mg / kg through the femoral vein. Thirty minutes after the administration, the rat was sacrificed by removing blood from the inferior aorta, and the tissue concentration was measured. Radioactivity in plasma and tissue was measured with a liquid scintillation counter after burning using a sample oxidizer.

The sample prepared by the above method is
99. Radioactive in 10% trichloroacetic acid solution.
It was observed that 5% was precipitated.

The results are shown in FIG. FIG. 13 is a graph showing the ratio (Kp value) between the brain concentration and the plasma concentration 30 minutes after administration for each sample. From FIG. 13, it can be seen that there is a possibility that sugar modification may enhance the transfer to the brain.

Example 53 p-Aminophenyl α-D-glucopyranoside (30
1) p-nitrophenyl α-D-glucopyranoside 500
To a solution of 7 mg of water in 15 ml of methanol, 75 mg of 10% palladium-carbon was added, and hydrogen gas was blown into the solution at room temperature for 16 hours. After removing the catalyst by filtration, the filtrate was concentrated to dryness under reduced pressure to obtain 490 mg of a crystalline residue. The residue was washed with a small amount of methanol to collect crystals, and the title compound (30
1) 210 mg (47%) was obtained. Table 9
To obtain compounds 301 to 308. The physical constants are shown in Table 9.

[0380]

[Table 9]

Example 54 p-Isothiocyanatophenyl α-D-glucopyranoside (309) Compound (301) 200 mg (0.737 mmol) 8
Thiophosgene 0.3% in 40 ml of 0% hydrous ethanol solution
2 ml (4.196 mmol) was added, and the mixture was stirred at room temperature for 2 hours. Nitrogen gas was blown into the reaction mixture to drive off excess thiophosgene, and sodium hydroxide solution was added to adjust the pH value to 6. The reaction solution was concentrated under reduced pressure to about 10 ml to obtain an aqueous solution containing the title compound (309). The aqueous solution of this compound was directly used for the condensation reaction with human serum albumin. Compounds 309 to 316 shown in Table 10 were obtained by the same method.

Example 55 1- [p- (α-D-glucopyranosyloxy) phenylamino] methanethioamide Human serum albumin (31
7) An aqueous solution containing the compound (309) was added to human serum albumin 1
12 mg (0.00167 mmol) of 0.08 M sodium borate-0.35 M potassium hydrogen carbonate buffer 60 ml
Was added dropwise. After leaving the reaction mixture at room temperature for 18 hours,
Dialysis membrane Spectrapol 2 (Molecular weight cutoff 12,0
00 to 14,000) against purified water at 4 ° C for 2
It was dialyzed for 4 hours. The dialyzed solution was taken out and freeze-dried to obtain 120 mg of the title compound (317). When the glucose content introduced was quantified by the phenol-sulfuric acid method, the glucose content (molar ratio) was 32. Compounds 317 to 324 shown in Table 10 were obtained by the same method. The physical constants of these compounds are shown in Table 11.

[0384]

[Table 10]

[0385]

[Table 11]

Test Example 6 Neoglycoproteins 318, 320, 322 and 324 having phenylthiourea as an arm obtained in Example 53
Was prepared as a specimen sample and HSA was prepared as a control sample. The sample was iodine-labeled by the same method as in Test Example 1 and a distribution experiment in animals was conducted. The results are shown in Table 12.

[0387]

[Table 12]

From Table 12, it is known that modification of albumin with sugar via phenylthiourea increases distribution clearance to bone marrow tissue. When it is mediated by phenylthiourea, the distribution clearance is large and the directivity to bone marrow tissue is remarkable.

The above shows that the glycosyl-protein derivative of the present invention having phenylthiourea as an arm is useful as a drug delivery carrier targeting bone marrow tissue.

Reference Example 57 2- (2-benzyloxyethoxy) ethoxyethanol (402) Triethylene glycol was added to a solution prepared by suspending 13.5 g (0.338 mol) of 60% sodium hydride in 300 ml of anhydrous dimethylformamide. (401) 50.0 g
(0.33 mol) was added dropwise. After stirring at room temperature for 1 hour, 56.0 g (0.33 mol) of benzyl bromide was added dropwise, and the mixture was further stirred at room temperature for 1 hour. After completion of the reaction, the solvent was distilled off under reduced pressure, the obtained residue was dissolved in 200 ml of ethyl acetate, and washed with 200 ml of water. The organic phase was dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure to give an oily crude product. This was purified by silica gel column chromatography [silica gel 70-230 mesh 500 g, solvent system: hexane / ethyl acetate (1: 1)] to obtain 27.5 g (34%) of the title compound (402).

¹ H-NMR (CDCl ₃ , δppm): 7.26 ~ 7.35 (5H, m), 4.57 (2H, s), 3.73 (2H, t, J = 4.5Hz), 3.66 ~ 3.71 (6H, m) , 3.63 ~ 3.65 (2H, m), 3.62 (2H, t, J = 4.5Hz), 2.25 (1H, br) IR (neat) cm ^-1 : 3450 (OH), 1099 (COC)

Reference Example 58 2- (2-benzyloxyethoxy) ethoxyethyl bromide (403) A solution of 36.0 g (0.15 mol) of compound (402) in 100 ml of anhydrous ether was added to phosphorus tribromide 14 under ice-cooling. 0.0 g
(0.052 mol) was added and stirred for 1 hour. 1 more
After stirring at room temperature for 100 hours, 100 ml of water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure to obtain a syrupy crude product. This was subjected to silica gel column chromatography [silica gel 70-230 mesh 250 g, solvent system:
Purification with hexane / ethyl acetate (9: 1)] afforded 13.6 g (36%) of the title compound (403).

¹ H-NMR (CDCl ₃ , δppm): 7.26 ~ 7.35 (7H, m), 4.58 (2H, s), 3.81 (2H, t, J = 7.0Hz), 3.66 ~ 3.71 (6H, m) , 3.61 ~ 3.63 (2H, m), 3.47 (2H, t, J = 6.7Hz) IR (neat) cm ^-1 : 1112 (COC)

Reference Example 59 Methyl 4- [2- (2-benzyloxyethoxy) ethoxy] -2-methoxycarbonylbutanoate (40
4) To a suspension of 60 g of sodium hydride 10.0 g (0.25 mol) in anhydrous dimethylformamide (300 ml) was added dimethyl malonate (33.0 g), and the mixture was stirred at 40 ° C. for 1 hour. To this was added 41.1 g (0.136 mol) of compound (403) all at once, and the mixture was further stirred at 40 ° C. for 6 hours and then left overnight at room temperature. Neutralize the reaction mixture with 10% hydrochloric acid,
The solvent was evaporated under reduced pressure and the obtained residue was partitioned with 100 ml of water and 200 ml of ethyl acetate. After separating the organic phase,
It was dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure to obtain a syrupy crude product. This was subjected to silica gel column chromatography [silica gel 70-230 mesh 250 g, solvent system: hexane / ethyl acetate (20:
1)], the title compound (404) was 36.8.
g (77%) was obtained.

¹ H-NMR (CDCl ₃ , δppm): 7.26 to 7.34 (5H, m), 4.56 (2H, s), 3.72 (3H, s), 3.52 (2H, t, J = 5.9Hz), 2.18 (2H, m) IR (neat) cm ^-1 : 1754 (C = O), 1735 (CO ₂ CH ₃ )

Reference Example 60 Methyl 4- [2- (2-benzyloxyethoxy) ethoxy] butanoate (405) Compound (404) 23.2 g (65.5 mmol) and sodium chloride 4.5 g (76.9 mmol). The water 4
ml and dimethylsulfoxide 80ml mixture, add 15
The mixture was heated and stirred at 0 to 160 ° C for 4 hours. After completion of the reaction, the solvent was distilled off under reduced pressure, the obtained residue was partitioned with 100 ml of water and 100 ml of ethyl acetate, and the organic phase was separated and dried over anhydrous sodium sulfate. The syrupy crude product obtained by distilling off the solvent under reduced pressure was subjected to silica gel column chromatography [silica gel 70-230 mesh 250 g,
Purification with solvent system: hexane / ethyl acetate (5: 1)] gave 17.0 g (87%) of the title compound (405).

¹ H-NMR (CDCl ₃ , δppm): 7.24 to 7.33 (5H, m), 4.57 (2H, s), 3.66 (3H, s), 3.63 to 3.68 (6H, m), 3.57 to 3.58 ( 2H, m), 3.50 (2H, t, J = 6.2Hz), 2.42 (2H, t, J = 7.3Hz), 1.90 (2H, m) IR (neat) cm ^-1 : 1738 (CO ₂ CH ₃ ) , 1113 (COC)

Reference Example 61 Methyl 4- [2- (2-hydroxyethoxy) ethoxy] butanoate (406) Compound (405) 16.6 g (56.0 mmol) in methanol 20 ml solution, 10% palladium carbon 2.0 g.
Was added and hydrogenated at room temperature for 4 hours. After the reaction was completed, the catalyst was filtered off, and the filtrate was concentrated to dryness under reduced pressure to obtain 11.3 g (99%) of the title compound 406 as a colorless oily substance.

¹ H-NMR (CDCl ₃ , δppm): 3.73 (2H, t, J = 4.5Hz), 3.68 (3H, s), 3.65 ~ 3.68 (2H, m), 3.57 ~ 3.63 (4H, m) , 3.51 (2H, t, J = 6.2Hz), 2.42 (2H, t, J = 7.3Hz), 1.92 (2H, m), 1.70 (1H, br) IR (neat) cm ^-1 : 3450 (OH) , 1738 (CO ₂ CH ₃ ), 1119 (COC) MS (EI) m / z: 207 (M + H ⁺ )

Example 56 2- [2- (3-methoxycarbonylpropyloxy)
Ethoxy] ethyl-2,3,4,6-tetra-O-benzyl-β-D-mannopyranoside (409) and 2-
[2- (3-Methoxycarbonylpropyloxy) ethoxy] ethyl-2,3,4,6-tetra-O-benzyl-α-D-mannopyranoside (408)

A solution of 577 mg (2.8 mmol) of the compound (406), 500 mg (1.79 mmol) of silver carbonate and 500 mg of drylite in 5 ml of methylene chloride was placed under a nitrogen stream.
After stirring at ℃ for 30 minutes, Koto, Morishima, Miyata,
and Zen, Bull. Chem. Soc. Jpn., 49, 2639 (1976), 2,3,4,6-tetra-O-benzyl-α / β.
580 mg of 2,3,4,6-tetra-O-benzyl-α-D-mannopyranosyl chloride (407) synthesized by blowing dry hydrogen chloride gas into a methylene chloride solution of -mannopyranosyl p-nitrobenzoate. 3 ml of methylene chloride solution (0.84 mmol) was slowly added dropwise.
Then, the mixture was stirred at an internal temperature of 0 to 5 ° C for 4 hours. After completion of the reaction, the reaction product was filtered through Celite, and the insoluble material was washed well with methylene chloride. The filtrate and washings were combined and the solvent was distilled off under reduced pressure to obtain 1.1048 g of a crude product. The crude product is flash chromatographed [silica gel 23
0 to 400 mesh 60 g, solvent system: ethyl acetate / hexane (2: 3)] to separate and purify the title compound (40
9) 520 mg (81.2%) and the title compound (40
8) 120 mg (18.8%) was obtained.

Compound (409) [α] ²⁴ _D −44.6 ° (C O.67, CHCl ₃ ) ¹ H-NMR (CDCl ₃ , δppm): 7.18 to 7.50 (20H, m, aromatic H), 4.85 to 4.97 (2H, AB-q, J = 12.5Hz, benzyl-CH ₂ ), 4.54 ~ 4.62 (2H, AB-q, J = 12Hz, benzyl-CH ₂ ), 4.50 ~ 4.90 (2H, AB-q, J = 12Hz, benzyl-CH ₂ ), 4.41 ~ 4.51 (2H, AB-q, J = 12Hz, benzyl-CH ₂ ), 4.44 (1H, s, H-1), 4.02 ~ 4.08 (1H, m, H-2 ), 3.64 ~ 3.93 (7H, m), 3.63 (3H, s, CO ₂ CH ₃ ), 3.40 ~ 3.64 (8H, m), 2.35 ~ 2.40 (2H, m, CH ₂ CO-), 1.80 ~ 1.95 ( 2H, m, CH ₂ CH ₂ CO-) IR (CHCl ₃ ) cm ^-1 : 1726 (CO ₂ CH ₃ ) MS (FAB) m / z: 751 (M + Na ⁺ ), 727 (M-1)

Compound (408) [α] ²⁴ _D + 13.9 ° (C O.70, CHCl ₃ ) ¹ H-NMR (CDCl ₃ , δppm): 7.18 to 7.50 (20H, m, aromatic H), 4.91 (1H , d, J = 1.71Hz, H-1), 4.68 to 4.75 (2H, AB-q, J = 12.5Hz, benzyl-CH ₂ ), 4.604 (2H, s, benzyl-CH ₂ ), 4.47 to 4.86 ( 2H, AB-q, J = 10.75Hz, benzyl-CH ₂ ), 4.51 ~ 4.65 (2H, AB-q, J = 12Hz, benzyl-CH ₂ ), 3.65 ~ 4.00 (8H, m), 3.63 (3H, s, CO ₂ CH ₃ ), 3.44 to 3.60 (8H, m), 2.30 to 2.40 (2H, m, CH ₂ CO-), 1.80 to 1.90 (2H, m, CH ₂ CH ₂ CO-) IR (CHCl ₃ ) cm ^-1 : 1725 (CO ₂ CH ₃ ) MS (FAB) m / z: 727 (M-1)

Example 57 2- [2- (3-methoxycarbonylpropyloxy)
Ethoxy] ethyl-β-D-mannopyranoside (41
0) 500 mg (0.686 mmol) of the compound (409) was dissolved in 10 ml of methanol, and 10% palladium-carbon 50 was added.
0 mg was added, and the mixture was vigorously stirred at room temperature under a hydrogen stream for 40 hours. After completion of the reaction, the catalyst was filtered off and the filtrate was concentrated to dryness under reduced pressure to give the title compound (410) 232.4 mg (92%)
was gotten.

Compound (410) [α] ²³ _D -22.3 ° (C 2.18, MeOH) ¹ H-NMR (CD ₃ OD, δppm): 4.53 (1H, s, H-1), 3.30 to 4.00 (14H, m), 3.62 (3H, s, CO ₂ CH ₃ ), 3.10 to 3.25 (2H, m), 2.37 (2H, t, J = 7.3Hz, CH ₂ CO-), 1.83 (2H, t, J = 7.3 Hz, CH ₂ CH ₂ CO-) IR (CHCl ₃ ) cm ^-1 : 3350 (OH), 1732 (CO ₂ CH ₃ ) MS (FAB) m / z: 391 (M + Na ⁺ )

Example 58 2- [2- (3-methoxycarbonylproxyoxy)
Ethoxy] ethyl 2,3,4-tri-O-acetyl-
β-L-fucopyranoside (412) HMFlowers et al. [Carbohydr. Res., 4, 189-195 (196
7)], 2,3,4-tri-O-acetyl-
α-L-fucopyranosyl bromide (411) 166 mg
To a solution of (0.469 mmol) in 10 ml of benzene, 100 mg (0.485 mmol) of the compound (406), 123 mg (0.485 mmol) of mercuric cyanide and 350 mg of powdered anhydrous calcium sulfate were added, and the mixture was heated at room temperature in an argon stream. It was stirred for 24 hours. After filtering the reaction solution, 5 ml of ethyl acetate was added to the filtrate, and the mixture was washed with 5 ml of water. The syrup obtained by drying over anhydrous magnesium sulfate and concentration under reduced pressure was purified by silica gel column chromatography (chloroform: methanol = 200: 1) to give the title compound (41
2) was obtained in 136 mg (58.6%).

Compound (412) [α] ²⁵ _D -2.9 ° (C 1.1, CHCl ₃ ) ¹ H-NMR (CDCl ₃ , δppm): 1.22 (3H, d, J = 6.4Hz, H-6), 1.99 , 2.06, 2.18 (9H, 3s, OAc), 2.42 (2H, t, J = 7.3Hz, C H ₂ , COOCH ₃ ), 3.68 (3H, s, OCH ₃ ), 4.76 (1H, d, J = 8.1 Hz, H-1) IR (NaCl) cm ^-1 : 1750, 1700

Example 59 2- [2- (3-methoxycarbonylpropyloxy)
Ethoxy] ethyl 2,3,4-β-L-fucopyranoside (413) 180 mg (0.376 mmol) of compound (412) was dissolved in 2 ml of methanol, and then 0.05 ml of 28% sodium methylate was added, and the mixture was stirred at room temperature. Stir for 2 hours. The reaction solution was neutralized with Amberlite IR-120B (H ⁺ ), the ion exchange resin was filtered off, and the filtrate was concentrated under reduced pressure to give the title compound (413) 109 mg (82.3%).

[Α] ²⁶ _D -2.1 ° (C 1.3, MeOH) ¹ H-NMR (CD ₃ OD, δppm): 1.23 (3H, d, J = 6.6Hz, H-6), 2.37 (2H, t , J = 7.6Hz, C H ₂ COOCH ₃ ), 3.67 (3H, s, OCH ₃ ), 4.19 (1H, d, J = 7.3Hz, H-1) IR (NaCl) cm ^-1 ： 3450, 1740, 1070

Example 60 2- [2- (3-methoxycarbonylpropyloxy)
Ethoxy] ethyl 3,4,6-tri-O-acetyl-
2-Azido-2-deoxy-α-D-mannopyranoside (414) and 2- [2- (3-methoxycarbonylpropyloxy) ethoxy] ethyl 3,4,6-tri-
O-acetyl-2-azido-2-deoxy-β-D-mannopyranoside (415)

Carbohydr. Res., 136, 153 (1985), 3,4,6-tri-O-acetyl-2-azide-
2-deoxy-α-D-mannopyranosyl bromide 9
30 mg (2.35 mmol) and methyl 4- [2- (2
-Hydroxyethoxy) ethoxy] butanoate (40
6) To a solution of 485 mg (2.35 mmol) in 20 ml of toluene, 700 mg of silver silicate and 500 mg of powdery molecular sieve 4A were added, and the mixture was stirred at room temperature for 16 hours. The reaction solution was filtered through Celite, the filtrate was concentrated to dryness under reduced pressure, and the obtained residue was purified by a Rover column (hexane: ethyl acetate = 1: 1) to give the title compound (4
14) 525 mg (43%) and the title compound (415) 1
43 mg (11.7%) was obtained.

Compound (414) [α] ²⁴ _D + 58.2 ° (C O.57, CHCl ₃ ) ¹ H-NMR (CDCl ₃ , δppm): 5.39 (1H, dd, J _3,4 = 10Hz, J _{2 , 3 = 4.0Hz, H-3} ), 5.32 (1H, t, J = 10Hz, H-4), 4.91 (1H, s, H-1), 4.25 (1H, dd, J 5,6a = 4.5Hz , J _{6a, 6b} = 12Hz, H-6a), 4.02 (1H, m, H-5), 2.40 (2H, t, J = 7.5Hz, -C H ₂ C O-), 2.10, 2.09, 2.04 ( each 3H, each s, 3 × OAc) IR (film) cm ^-1 : 2110, 1747, 1438, 1369, 1230, 1049

Compound (415) [α] ²⁴ _D -60.1 ° (C O.9, CHCl ₃ ) ¹ H-NMR (CDCl ₃ , δppm): 5.24 (1H, t, J _3,4 = 9.5Hz, H -4), 4.99 (1H, dd , J 2,3 = 3.5Hz, H-3), 4.79 (1H, s, H-1), 4.25 (1H, dd, J 5,6a = 5.5Hz, J 6a _{, 6b} = 12Hz, H-6a), 4.00 (1H, m, H-5), 2.40 (2H, t, J = 7.5Hz, -C H ₂ C O-), 2.10, 2.08, 2.03 (each 3H, each s, 3 × OAc) IR (film) cm ^-1 : 2112, 1743, 1371, 1236, 1055

Example 61 2- [2- (3-methoxycarbonylpropyloxy)
Ethoxy] ethyl 2-azido-2-deoxy-α-D
-Mannopyranoside (416) To a solution of 525 mg (1.01 mmol) of compound (415) in 10 ml of anhydrous methanol, 1M sodium methoxide-
0.25 ml of methanol solution was added, and the mixture was left at room temperature for 4 hours. Ion exchange resin Amberlite IR-12 for reaction liquid
After neutralizing by adding 0B (H ⁺ type), the resin was filtered off and washed with a small amount of methanol. The filtrate and washings were combined, and the solvent was distilled off under reduced pressure to give the syrup-like title compound (41
6) 154 mg was obtained.

Example 62 2- [2- (3-methoxycarbonylpropyloxy)
Ethoxy] ethyl 2-acetamido-2-deoxy-α
-D-mannopyranoside (417) Compound (416) 150 mg (0.381 mmol) in ethanol 4 ml solution, nickel chloride hexahydrate 380 mg
After adding 0.1 ml of a solution of (1.6 mmol) in 10 ml of ethanol, 43 mg of sodium borohydride
4 ml was added to a solution of (1.143 mmol) in ethanol with stirring. After stirring at room temperature for 30 minutes, acetic acid was added to the reaction solution to neutralize it. Then, 0.5 ml of acetic anhydride was added and the mixture was left at room temperature for 1 hour. The reaction mixture was concentrated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform: methanol = 7: 1) to obtain 157 mg of the title compound (417).

[Α] ²⁶ _D + 26.4 ° (C 0.62, MeOH) ¹ H-NMR (CD ₃ OD, δppm): 4.72 (d, 1H, J = 1Hz, H-1), 4.31 (dd, 1H, J _1,2 = 1Hz, J _2,3 = 4.7Hz), 2.01 (s, 3H, NAc) IR (KBr) cm ^-1 : 3446, 1730, 1660, 1550, 1132, 1068

Example 63 Neoglycoprotein (419) To a solution of 85 mg (0.241 mmol) of compound (413) in 0.8 ml of ethanol was added 0.24 of hydrazine hydrate.
ml was added, and the mixture was stirred at room temperature for 24 hours. The reaction solution was concentrated under reduced pressure, 1 ml of water was added, and the mixture was further concentrated under reduced pressure to give 2- [2- (3-hydrazinocarbonylpropyloxy) ethoxy] ethyl β-L-fucopyranoside 85 mg.
I got To this, add 3 ml of DMF and dissolve, then at -25 ° C.
After cooling to 0.14 ml of 4N hydrogen chloride in dioxane was added. Furthermore, at the same temperature, t-butyl nitrite 35.5 mg
(0.241 mmol) of DMF (0.2 ml) was added and stirred for 30 minutes, and then 16.8 mg of sulfamic acid in DM
Add 0.2 ml F solution and stir for 15 minutes
A reaction solution containing the corresponding acid azide was obtained. 332 mg (4.8 mg) of human serum albumin (HSA) cooled to 0 ° C.
2 × 10 ⁻³ mmol) borate buffer (0.08 M Na ₂
B ₄ O ₇ , 0.35M KHCO ₃ ) (20 ml) and the mixture was stirred at the same temperature for 24 hours. After dialysis at 4 ° C. overnight, lyophilization gave powder of fucose-modified albumin (419) 343 mg. It was found by the phenol-sulfuric acid method that the number of moles of sugar chains bound to HSA / mol was 28.

A list of neoglycoprotein and the intermediates used is shown in Table 13. The introduced sugar content was measured by the Elson-Morgan method after hydrolysis with a phenol-sulfuric acid method in the case of neutral sugar and with 4N hydrochloric acid in the case of hexosamine. Table 14 shows the physical constants of these neoglycoproteins.

[0419]

[Table 13]

[0420]

[Table 14]

Test Example 7 Neoglycoproteins 418, 419 and 420 having the ethylene oxide arm obtained in Example 54 as the sample were prepared, and HSA was prepared as the control sample. The sample was iodine-labeled by the same method as in Test Example 1 and a distribution experiment in animals was conducted. The results are shown in Table 15.

[0422]

[Table 15]

From Table 15, it is known that modification of albumin with sugar through ethylene oxide increases distribution clearance to bone marrow tissue. When mediated by ethylene oxide, the blood concentration persistence after intravenous administration is almost comparable to that in HSA, and therefore, it is characterized in that it is slowly distributed to bone marrow tissue.

The above shows that the glycosyl protein derivative of the present invention having ethylene oxide as an arm is useful as a drug delivery carrier targeting bone marrow tissue.

[Brief description of drawings]

FIG. 1 is a graph showing HSA uptake activity.

FIG. 2 is a graph showing the uptake activity of compound 236.

FIG. 3 is a graph showing the uptake activity of compound 226.

FIG. 4 is a graph showing the uptake activity of compound 238.

FIG. 5 is a graph showing the uptake activity of compound 246.

FIG. 6 is a graph showing the uptake activity of compound 245.

FIG. 7 is a graph showing the uptake activity of compound 242.

FIG. 8 is a graph showing the uptake activity of compound 245.

FIG. 9 is a graph showing the uptake activity of compound 242.

FIG. 10 is a graph showing a comparison of specific uptake activities of (Man β-O (CH ₂ ) ₈ CO) _m -HAS having different m numbers.

FIG. 11 is a graph showing a comparison of specific uptake activities of (Fuc β-O (CH ₂ ) ₈ CO) _m -HAS having different m numbers.

FIG. 12 is a graph showing the inhibitory effect of various neoglycoproteins on compound 245.

FIG. 13 is a graph showing the ratio (Kp value) between brain concentration and plasma concentration of various neoglycoproteins.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical indication C07K 14/52 8318-4H 14/76 8318-4H // C07H 15/04 F (72) Inventor Takashi Kato 1-7-1, Higashi, Kashiwa-shi, Chiba 104, Grand Ari 104 (56) References JP-A-63-152393 (JP, A)

Claims (5)

[Claims] 1. A formula (I): _{[R-O- (CH 2) n} -CO- ] _m -Z (I) (wherein, R represents a glycosyl group, Z is expressed protein, n
Represents 3 to 14 and m represents 10 to 50), and is a drug delivery carrier containing the glycosyl-protein derivative. 2. A carrier for drug delivery to bone marrow, which comprises the glycosyl-protein derivative of claim 1. 3. A carrier for drug delivery to the brain containing the glycosyl-protein derivative according to claim 1, wherein m is 20 to 50. 4. The glycosyl group is a xylopyranosyl group,
Mannopyranosyl group, fucopyranosyl group, galactopyranosyl group, 2-acetamido-2-deoxy-fucopyranosyl group, 2-acetamido-2-deoxy-mannopyranosyl group, 2-acetamido-2-deoxy-galactopyranosyl group, mannopyranosyl- Mannopyranosyl group, (2-acetamido-2-deoxy-mannopyranosyl) -mannopyranosyl group, (2-acetamido-2
-Deoxy-glucopyranosyl) -mannopyranosyl group, fucopyranosyl- (2-acetamido-2-deoxy-glucopyranosyl) group, galactopyranosyl- (2
-Acetamido-2-deoxy-glucopyranosyl)
Group, galactopyranosyl- (2-acetamido-2-deoxy-mannopyranosyl) group, galactopyranosyl-
A glucopyranosyl group, a di (2-acetamido-2-deoxy-glucopyranosyl) -mannopyranosyl group or a di (galactopyranosyl) -2-acetamido-2-deoxy-glucopyranosyl group, which is any one of claims 1 to 3. The drug delivery carrier according to item 1. 5. The drug delivery carrier according to any one of claims 1 to 4, wherein the protein is albumin, cytokines or enzymes.