JP2589435B2 - Sugar-modified polylysine derivative - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel sugar-modified polylysine derivative useful as an organ-directing drug delivery carrier.

[0002]

2. Description of the Related Art Polylysine is used as a drug delivery carrier.
For example, the following literature mainly describes the delivery of anticancer drugs.
It has been studied as a body. 1) L.J. Arnold, Jr. et al., Targeted Drugs (Goldberg,
E.P., ed,) J. Wiley & Sons, 89-112 (1983) 2) H.J.P. Ryser et al., Proc. Natl. Acad. Sci., USA7
Five, 3867-3870 (1987) 3) Y. Morimoto et al., J. Pharmacobiodyn,9, 688-698 (198
Four)

[0003] In the above Reference 1, an example in which daunomycin, methotrexate, and 6-aminonicotinamide were bonded to polylysine, in Reference 2, an example in which methotrexate was bonded to polylysine, and in Reference 3, melphalan was also bonded. It is shown. Further, attempts to modify polylysine with sugar and use it as a drug delivery carrier are also described in, for example, the following literature. 4) D, Derrien et al., Glycoconjugate, 6 , 241-255 (1989) 5) L. Fumie et al., FEBS Letters, 203 , 203-206 (1986) 6) CAHoppe et al., Biochemistry, 23 , 1723-1730 (1984)

[0004] Reference 4 discloses an attempt to use a polylysine modified with mannose as a drug carrier and to bind a muramyl dipeptide to make it an activator of macrophages. In Document 5, similarly, the one modified with galactose was used as a liver-directed drug delivery carrier,
An example in which arabinofuranosyl adenine monophosphate is bound is shown. Reference 6 discloses an example in which polylysine modified with mannose and galactose via sulfur is bound to a receptor of lung macrophages.

[0005] As shown in these examples, in order to allow polylysine to bind an anticancer agent or an antitumor agent to enable selective transport to cancerous tissues or tumor cells, a sugar that recognizes these tissues or cells is used. Is generally known, and it was therefore predicted that sugar-modified polylysine could be used as a drug delivery carrier imparted with organ tropism. However, as shown in the experimental examples described below, sugar-modified polylysine is unstable in blood and causes nonspecific organ adhesion. This is considered to be due to the fact that a free ε-amino group to which no sugar or drug is bound remains in polylysine, and remains as a problem to be solved in drug delivery technology.

[0006]

In view of the above situation in the prior art, the present inventors have made various studies and found that a part of the free ε-amino group in the sugar-modified polylysine contains an ethylene glycol chain. The present inventors have found that the modification with a group stabilizes in blood and avoids non-specific organ attachment, thereby completing the present invention. That is, an object of the present invention is to provide a technique for improving adhesion to a non-target organ which inhibits its usefulness when using a sugar-modified polylysine as a drug delivery carrier to a specific organ.

Hereinafter, the present invention will be described in detail. The substance of the present invention comprises a structural unit (A) represented by the following formula (I):
A sugar-modified polylysine derivative comprising (B) and (C),

[0008]

Embedded image

In the formula, R ¹ is a glycosyl group, and X is the following formula (I
I)

[0010]

Embedded image

R ² represents the following formula (III) or (IV):

[0012]

Embedded image

R ³ represents a hydrogen atom or a drug residue. p,
q and γ represent the numbers of the structural units (A), (B) and (C), and the numbers are 1 to 200, 10 to 300 and 1 respectively.
-20.

Formula (I) shows that the sugar-modified polylysine derivative of the present invention is composed of three different lysine units. The first unit (A) is a lysine unit modified by a sugar, the second unit (B) is a lysine unit modified by a group containing an ethylene glycol chain, and the third unit (C) is not yet modified. Modified or drug modified lysine units. Further, the formula (I) means that each of these three types (A), (B) and (C) contains only p, q and r units in one molecule. Further, the description of the formula (I) is based on the units (A), (B) and (C).
Means that they are randomly copolymerized. The first unit (A) is indispensable for functioning the organ tropism in the substance of the present invention, and the second unit (B) is the stability of the substance of the present invention in blood and non-specific organs. It is essential to avoid adhesion to the ε-amino group, even when a unit having a free ε-amino group which is not used as a drug carrier among the third units (C) remains. This has the effect of not causing adverse effects.

R ¹ in the substance of the formula (I) represents a glycosyl group, including those whose hydroxyl group is protected or unprotected by an acetyl group. Examples of glycosyl groups include, for example, D-mannopyranosyl, D-galactopyranosyl, L-fucopyranosyl, xylopyranosyl, 2-acetamido-2-deoxy-D-galactopyranosyl, 2-acetamido-2-
Deoxy-D-mannopyranosyl, 2-acetamide-2
-Deoxy-D-glucopyranosyl, 4-galactopyranosyl-D-glucopyranosyl, 4-galactopyranosyl-2-acetamido-2-deoxy-D-glucopyranosyl, 2-acetamido-2-deoxy-6-fucosyl-
D-glucopyranosyl, 6-mannopyranosyl-D-mannopyranosyl, 3,6-di-mannopyranosyl-D-
Mannopyranosyl can be mentioned.

The sugar content of the substance of the formula (I) can be determined by colorimetry by the phenol-sulfuric acid method or the Nilsson-Morgan method. The optical isomerism in the lysine unit is not particularly limited, and accordingly, poly-D-lysine,
Any of poly-L-lysine and poly-DL-lysine may be used. X is a spacer which is between the sugar and the ε-amino group of lysine and connects them, and in the present invention, is a linear alkylene group or an alkylene group containing an ethylene glycol chain. R ² is a protecting group for a free ε-amino group of a lysine unit, which corresponds to an alkyl group (III or IV) containing an ethylene glycol chain in the present invention, and is expected to enhance water solubility, It has the effect of increasing stability in the body and avoiding non-specific adhesion to organs.

R ³ represents a hydrogen atom or a drug residue (a drug or a drug-attached spacer bonded to an ε-amino group is generally referred to as a drug residue). In the drug residue, examples of the terminal group of the portion bonded to the amino group include a carbonyl group, a sulfonyl group, and a phosphonyl group. When the sugar-modified polylysine derivative (I) needs to be subjected to iodine labeling for use in animal experiments, R
³ has a hydroxyphenylethylcarbonyl group bonded thereto.

The third unit (C) includes not only a case where R ³ is occupied by only a unit of a hydrogen atom or a drug residue, but also a case where R ³ is occupied by a mixture of these units. The molecular weight of the substance of formula (I) is not limited, but is usually between 5,000 and 150,000. The measurement is
For example, it is possible by a viscosity method or a LALLS method. The production of the sugar-modified polylysine derivative of the formula (I) is carried out, for example, using commercially available polylysine as a starting material as follows. First, R ¹ O—X—CO as a sugar chain is added to polylysine.
In order to introduce a group, for example, an acid azide of R ¹ O—X—COOH is prepared and reacted, or R ¹ O—X—COOH is reacted using a suitable condensing agent. Then R ³
For example, when R ³ is a drug residue having a certain carbonyl group, R ³ OH is reacted with hydroxysuccinimide using a condensing agent such as DCC. . Finally, to introduce an R ² CO group, for example, an oxysuccinimide ester of R ² CO is reacted. Normally, as described above, the R ² CO group is introduced after the introduction of the drug residue of R ³ , but the reverse is also possible. In this case, the oxysuccinimide ester of R ² CO is reacted again.

The above R ¹ O—X—COOH is produced as follows. First, a hydroxyl group of the glycosyl group of R ¹ is protected with an acetyl group, a benzyl group, or the like, or a glycosyl halide in which an amino group is protected in the form of an azide group is HO
An oxyacid represented by -X-COOH or an ester thereof is reacted. The reaction is carried out according to a conventional method using a condensing agent commonly used for glycosylation. Finally, the acetyl group is eliminated with sodium methylate, and the benzyl group is eliminated with the palladium-carbon reduction method, or the azide group is converted to an amino group with sodium borohydride in the presence of nickel chloride hexahydrate, and If necessary, the amino group is acetylated with acetic anhydride. When an oxyester is used, the desired compound can be obtained by hydrolysis with an alkali.

[0020]

The substance of the present invention is stable in blood and can avoid non-specific adhesion to organs, as shown in the experimental examples described later.

[0021]

The present invention will be described more specifically with reference to examples and reference examples. The reaction steps corresponding to the examples are shown in Reaction Formulas 1 to 14. The compound numbers in the reaction formulas correspond to the compound numbers described in Reference Examples and Examples.

[0022]

Embedded image

[0023]

Embedded image

[0024]

Embedded image

[0025]

Embedded image

[0026]

Embedded image

[0027]

Embedded image

[0028]

Embedded image

[0029]

Embedded image

[0030]

Embedded image

[0031]

Embedded image

[0032]

Embedded image

[0033]

Embedded image

[0034]

Embedded image

[0035]

Embedded image

[0036]

Embedded image

[0037]

Embedded image

[0038]

Embedded image

[0039]

Embedded image

[0040]

Embedded image

[0041]

Embedded image

Reference Example 1 2- [2- (2-benzyloxyethoxy) ethoxy] ethanol (2) 60% in 300 ml of anhydrous N, N-dimethylformamide
To a solution of 13.5 g (0.338 mol) of sodium hydride suspended was added 50.0 g of triethylene glycol (1).
(0.33 mol) was added. After stirring for 1 hour at room temperature,
56.0 g (0.33 mol) of benzyl bromide was added, 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, and the obtained residue was dissolved in 200 ml of ethyl acetate.
Washed with 200 ml of water. The organic phase was dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure to obtain an oily crude product.
This was purified by silica gel column chromatography (n-hexane: ethyl acetate = 1: 1) to obtain 27.5 g (34%) of the title compound (2). 2: Oil IR (film) ν _max cm ^-1 : 3450, 1099 ¹ H-NMR (CDCl ₃ ): δ 2.25 (1H, br) 3.62 (2H, t, J = 4.5 Hz) 3.63-3.65 (2H, m ) 3.66-3.71 (6H, m) 3.73 (2H, t, J = 4.5Hz) 4.57 (2H, s, OC H ₂ ) 7.26-7.35 (5H, m, aromatic H)

REFERENCE EXAMPLE 2 2- [2- (2-Benzyloxyethoxy) ethoxy] ethyl bromide (3) A solution of 36.0 g (0.15 mol) of compound (2) in 100 ml of anhydrous ether was added under ice-cooling. 14.0 g of phosphorus bromide (5
2 mmol) and stirred for 1 hour. After further stirring at room temperature for 1 hour, 100 ml of water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase is dried over anhydrous sodium sulfate,
Concentration to dryness under reduced pressure gave a crude product in the form of a syrup. This was purified by silica gel column chromatography (n-hexane: ethyl acetate = 9: 1) to obtain 13.6 g (36%) of the title compound (3). 3: oily IR (film) ν _max cm ^-1 : 1112 ¹ H-NMR (CDCl ₃ ): δ 3.47 (2H, t, J = 6.7 Hz) 3.61-3.63 (2H, m) 3.66-3.71 (6H, m ) 3.81 (2H, t, J = 7.0Hz) 4.58 (2H, s, OC H ₂ ) 7.26-7.35 (5H, m, aromatic H)

Reference Example 3 Methyl 4- [2- (2-benzyloxyethoxy) ethoxy] -2-methoxycarbonyl-1-butanoate (4) 60 g of sodium hydride 10.0 g (0.25 mol) of anhydrous N , N-dimethylformamide in 300 ml suspension,
33.0 g of dimethyl malonate was added, and the mixture was stirred at 40 ° C for 1 hour. To this solution was added 41.1 g of compound (3) (0.13 g).
6 mol) at once, and after further stirring at 40 ° C. for 6 hours,
Left at room temperature overnight. The reaction mixture was neutralized with 10% hydrochloric acid, and the solvent was distilled off under reduced pressure.
And 200 ml of ethyl acetate. After separating the organic phase,
After drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure to obtain a syrupy crude product. This was subjected to silica gel column chromatography (n-hexane: ethyl acetate = 2
0: 1) to give 36.8 g of the title compound (4) (7
7%). 4: Oil IR (film) ν _max cm ^-1 : 1754, 1735 ¹ H-NMR (CDCl ₃ ): δ 2.18 (2H, m) 3.52 (2H, t, J = 5.9 Hz) 3.72 (6H, s, COOMe × 2) 4.56 (2H, s, OC H ₂ ) 7.26-7.34 (5H, m, aromatic H)

Reference Example 4 Methyl 4- [2- (2-benzyloxyethoxy) ethoxy] -1-butanoate (5) 23.2 g (65.5 mmol) of compound (4) and 4.5 g of sodium chloride (76. 9 mmol) was added to a mixture of 4 ml of water and 80 ml of dimethyl sulfoxide.
The mixture was heated and stirred at 60 ° C. for 4 hours. After completion of the reaction, the solvent was distilled off under reduced pressure, and the obtained residue was partitioned with 100 ml of water and 100 ml of ethyl acetate. The organic phase was separated and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting syrupy crude product was purified by silica gel column chromatography (n-hexane: ethyl acetate = 5: 1) to give 17.0 g of the title compound (5) ( 87%). 5: Oil IR (film) ν _max cm ^-1 : 1738, 1113 ¹ H-NMR (CDCl ₃ ): δ 1.90 (2H, m) 2.42 (2H, t, J = 7.3 Hz) 3.50 (2H, t, J = 6.2Hz) 3.57-3.58 (2H, m) 3.63-3.68 (6H, m) 3.66 (3H, s, COOMe) 4.57 (2H, s, OC H ₂ ) 7.24-7.33 (5H, m, aromatic H)

Reference Example 5 Methyl 4- [2- (2-hydroxyethoxy) ethoxy] -1-butanoate (6) A 10% palladium solution was added to a solution of 16.6 g (56.0 mmol) of compound (5) in 20 ml of methanol. 2.0 g of carbon was added and hydrogenated at room temperature for 4 hours. After completion of the reaction, the catalyst was removed by filtration, and the filtrate was concentrated to dryness under reduced pressure to obtain 11.3 g (99%) of the title compound (6) as a colorless oil. 6: Oil IR (film) ν _max cm ^-1 : 3450, 1738, 1119 MS (EI): m / z 207 (M + H ⁺ ) ¹ H-NMR (CDCl ₃ ): δ 1.70 (1H, br) 1.92 (2H, m) 2.42 (2H, t, J = 7.3Hz, C H ₂ COOMe) 3.51 (2H, t, J = 6.2Hz) 3.57-3.63 (4H, m) 3.65-3.68 (2H, m) 3.68 ( 3H, s, COOMe) 3.73 (2H, t, J = 4.5Hz)

Reference Example 6 2-methoxyethoxyacetic acid benzyl ester (8) 8 g of 60% sodium hydride was suspended in 100 ml of dry tetrahydrofuran, and 16 g of ethylene glycol monomethyl ether (7) (0) .
21 mol) was slowly added dropwise. After stirring for 1 hour at room temperature, the solution was added to 50 g of bromoacetic acid benzyl ester.
(0.218 mol). The reaction was completed by stirring at room temperature for 3 hours. The residue obtained by distilling off tetrahydrofuran under reduced pressure was extracted with ethyl acetate, washed with water, washed with saturated saline and dried over anhydrous magnesium sulfate. filtration,
The solution was distilled off under reduced pressure to obtain 50.8 g of a crude product.
This was purified by silica gel column chromatography (n-hexane: ethyl acetate = 1: 1) to obtain 27.9 g of a colorless transparent oil. This was further distilled under reduced pressure to obtain 17.0 g (36%) of a purified product of the title compound (8). 8: oily bp _1.0 113-118 ° C IR (film) ν _max cm ^-1 : 1755, 1278, 1200, 1120 ¹ H-NMR (CDCl ₃ ): δ 3.34 (3H, s, OMe) 3.54-3.58 (2H, m, OC H ₂ ) 3.68-3.72 (2H, m, OC H ₂ ) 4.17 (2H, s, OC H ₂ COOCH ₂ Ph) 5.16 (2H, s, OC H ₂ Ph) 7.25-7.40 (5H, br s , aromatic H)

Reference Example 7 Ethyl 2-methoxyethoxyacetate (9) Benzyl ester (8) 12.5 g (55.7 mmol) was dissolved in ethanol (50 ml), and 10% palladium-
Add 1.5g of carbon catalyst, and use the Pearl equipment for about 50ps
The reaction was performed for 18 hours under a medium pressure hydrogen stream of i. After completion of the reaction, the reaction mixture was filtered to remove the catalyst, and the filtrate was distilled off under reduced pressure.
35 g of the product were obtained. This was purified by distillation under reduced pressure to give the title compound (9) as a colorless and transparent oily substance, 4.9.
2 g (54.4%) were obtained. 9: oily bp _1.0 85-90 ° C IR (film) ν _max cm ^-1 : 1755, 1200, 1120 ¹ H-NMR (CDCl ₃ ): δ 1.29 (3H, t, J = 7.2 Hz, CH ₂ CH ₃ ) 3.39 (3H, s, OMe) 3.58-3.62 (2H, m, OC H ₂ ) 3.72-3.76 (2H, m, OC H ₂ ) 4.15 (2H, s, OC H ₂ COOEt) 4.22 (2H, q, J = 7.2Hz, OC H ₂ CH ₃ )

REFERENCE EXAMPLE 8 12 g (74 mmol) of ethyl 2-methoxyethoxyacetic acid (10) (9) was added dropwise to a solution of 3.6 g (90 mmol) of sodium hydroxide in 40 ml of water, and the mixture was allowed to stand at room temperature. And stirred for 18 hours. After completion of the reaction, the aqueous solution was concentrated under reduced pressure, and 50 ml of water was added again to the obtained residue to dissolve the residue.
Neutralized with -120B (H ⁺ type). After removing the resin by filtration, water was distilled off under reduced pressure, the obtained oil was dissolved in chloroform, dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure to give the title carboxylic acid ( 8) (80%) of 10) as a viscous oil. 10: Syrup IR (film) ν _max cm ^-1 : 3500, 3200, 1730, 1600 ¹ H-NMR (D ₂ O): δ 3.39 (3H, s, OMe) 3.64-3.68 (2H, m, OC H ₂ ) 3.70-3.74 (2H, m, OC H ₂ ) 4.08 (2H, s, OC H ₂ COOH)

Reference Example 9 8- (methoxycarbonyl) octyl-2,3,4,6-tetra-0-benzyl-β-
D-mannopyranoside (13) 8- (methoxycarbonyl) octanol (12) 10
g (53 mmol), 10 g (36.3 mmol) of silver carbonate, 10 g of anhydrous calcium sulfate and 150 ml of dry methylene chloride were stirred at room temperature for 30 minutes in a nitrogen stream, and then described in the literature [Koto, Morishima, Miyata and Zen, B
2,3,4,6-tetra-O-benzyl-α / β-D- obtained by the method of ull. Chem. Soc. Jpn., 49 , 2639 (1976)].
A dry hydrogen chloride gas was blown into a methylene chloride solution of mannopyranosyl P-nitrobenzoate, and a solution of 12.97 g (22.4 mmol) of chloride (11) synthesized in 100 ml of dry methylene chloride was slowly added dropwise. 1 at room temperature
After stirring and reacting for 8 hours, the reaction mixture was filtered using celite, and the filtrate was distilled off under reduced pressure to obtain 22.53 g of a crude product. This was purified by silica gel chromatography (n-hexane: ethyl acetate = 3: 1) to give the title compound (13) 1
2.25 g (76.8%) were obtained. 13: syrup ^{_{[α] D 25 - 38.1 °}} (c 1.39, CHCl 3) 1 H-NMR (CDCl 3): δ 1.20-1.40 (8H, br, C H 2 × 4) 1.58-1.70 (4H, m , C H ₂ × 2) 2.29 (2H, t, J = 7.57 Hz, C H ₂ COOMe) 3.39 (1H, dt, Ha of OCH ₂ ) 3.42-3.46 (1H, m, H-5) 3.49 (1H, dd, H-4) 3.65 (3H, s, COO Me ) 3.73 (1H, dd, J = 6.11Hz, 10.75Hz, Ha of C H ₂ OBzl) 3.80 (1H, dd, J = 1.7Hz, 10.75Hz, Hb of C H ₂ OBzl) 3.85 (1H, t, J = 9.53Hz, H-3) 3.89 (1H, d, J = 2.93Hz, H-2) 3.96 (1H, dt, Hb of OCH ₂ ) 4.36 ( 1H, s, H-1) 4.43, 4.49 (2H, AB-q, J = 11.97Hz, OC H ₂ Ph) 4.59, 4.61 (2H, AB-q, J = 12.5Hz, OC H ₂ Ph) 4.52, 4.90 (2H, AB-q, J = 11.0Hz, OC H ₂ Ph) 4.87, 4.98 (2H, AB-q, J = 12.46Hz, OC H ₂ Ph) 7.10-7.50 (20H, m, aromatic H)

Reference Example 10 8- (methoxycarbonyl)
Octyl-β-D-mannopyranoside (14) A mixture of 1.7 g (2.39 mmol) of compound (13), 20 ml of methanol and 500 mg of 10% palladium-carbon was hydrogenated with a Parr apparatus under a hydrogen stream of 50 psi. went. After reacting for 20 hours, the catalyst was removed by filtration, and the filtrate was distilled off under reduced pressure to obtain 693 mg (83%) of the title compound (14) as a colorless solid. 14: mp94-96 ℃ (2-PrOH- isopropyl ^{_{ether) [α] D 26 - 34.3}} ° (c 0.8, MeOH) 1 H-NMR (CD 3 OD): δ 1.20-1.40 (8H, br, C H 2 × 4) 1.50-1.70 (4H, br, C H ₂ × 2) 2.29 (2H, t, J = 7.30Hz, C H ₂ COOMe) 3.15-3.20 (1H, m) 3.40-3.44 (1H, m) 3.48 -3.56 (2H, m) 3.63 (3H, s, COOMe) 3.66-3.72 (1H, m) 3.80-3.90 (1H, m) 4.46 (1H, d, J = 0.73Hz, H-1)

Reference Example 11 2- [2- (3-methoxycarbonylpropyloxy) ethoxy] ethyl-3,4
6-tri-O-acetyl-2-azido-2-deoxy-
α-D-galactopyranoside (18) and 2- [2-
(3-methoxycarbonylpropyloxy) ethoxy]
Ethyl-3,4,6-tri-O-acetyl-2-azido-2-deoxy-β-D-galactopyranoside (19) By the same method as described in Reference Example 24 below,
3,4,6-tri-O-acetyl-2-azido-2-deoxy-α-D-galactopyranosyl bromide (17)
2.65 g (6.72 mmol) and methyl 4- [2-
1.386 g (6.72 mmol) of (2-hydroxyethoxy) ethoxy] -1-butanoate (6) were condensed with toluene using silver silicate in the presence of a powdered molecular sieve 4A to give the title compound (18). And (1
9) at 60 mg (1.7%) and 2.01 g (5
7.6%). 18: syrup ^{_{[α] D 24 - 55.5 °}} (c 0.95, CHCl 3) IR (CHCl 3) ν max cm -1: 2118, 1745, 1440, 1371, 1076 1 H-NMR (CDCl 3): δ 1.90 (2H, m, OC H ₂ ) 2.04 (3H, s, OAc) 2.09 (3H, s, OAc) 2.18 (3H, s, OAc) 2.40 (2H, m, C H ₂ CO) 3.49 (2H, t, J = 6.5Hz, OC H ₂ ) 3.57 (2H, m, OC H ₂ ) 3.63 (2H, m, OC H ₂ ) 3.67 (3H, s, COOMe) 4.02 (1H, dt, OC H ₂ ) 4.23 (2H , m, OC H ₂ ) 4.74 (1H, d, J _1,2 = 6.5Hz, H-1) 5.04 (1H, t, J _2,3 = 4Hz, H-3) 5.29 (1H, dd, J _{3 , 4 = 3.5Hz, J 4,5 =} 0.5Hz, H-4) 19: syrup ^{_{[α] D 20 - 13.7 °}} (c 2.06, CHCl 3) IR (CHCl 3) ν max cm -1: 2118, 1747, 1439, 1369, 117
4, 1078 ¹ H-NMR (CDCl ₃ ): δ 1.87 (2H, m, OC H ₂ ) 2.11 (3H, s, OAc) 2.38 (2H, t, J = 7 Hz, C H ₂ CO) 3.47 (2H, t, J = 6.5Hz, OC H ₂ ) 3.55 (2H, m, OC H ₂ ) 3.64 (3H, s, COOMe) 3.82 (2H, m, OC H ₂ ) 4.00 (1H, dt, OC H ₂ ) 4.10 (2H, m, OC H ₂ ) 4.45 (1H, d, J _1,2 = 8Hz, H-1) 4.73 (1H, dd, J _2,3 = 11Hz, J _3,4 = 3.5Hz, H-3 ) 5.29 (1H, dd, J _3,4 = 3.5Hz, J _4,5 = 0.5Hz, H-4)

Reference Example 12 2- [2- (3-methoxycarbonylpropyloxy) ethoxy] ethyl-2-azido-2-deoxy-β-D-galactopyranoside (2
0) To a solution of 2.0 g (3.85 mmol) of compound (19) in 40 ml of methanol, 1.54 ml of a 1M sodium methoxide-methanol solution was added in the same manner as described in Reference Example 25 below to remove O- Acetylation afforded 1.47 g (97%) of the title compound (20). 20: syrup-like [α] _D ²³ + 18.3 ° (c 0.82, MeOH) IR (film) ν _max cm ^-1 : 3400, 2116, 1727, 1570, 1440,
1354, 1259, 1070

Reference Example 13 2- [2- (3-ethoxycarbonylpropyloxy) ethoxy] ethyl-2-acetamido-2-deoxy-β-D-galactopyranoside (21) and 2- [2- (3 -Methoxycarbonylpropyloxy) ethoxy] ethyl-2-acetamide-2-
Deoxy-β-D-galactopyranoside (22) In a manner similar to that described in Reference Example 26 below, a solution of 1.45 g (3.686 mmol) of compound (20) in 15 ml of ethanol was added with 0.16 mM chloride. 0.69 ml of an ethanol solution of nickel hexahydrate was added, and the azide group was reduced to an amino group with 418 mg (11 mmol) of sodium borohydride. After N-acetylation with 3 ml of acetic anhydride, the residue was purified by silica gel column chromatography [chloroform: methanol: water = 7: 3: 1 (lower layer)] to give 311 of the title compounds (21) and (22), respectively.
mg (21%), 560 mg (37.3%). 21: Amorphous powder [α] _D ²⁰ -2.6 ° (c 0.50, MeOH) IR (KBr) ν _max cm ^-1 : 3350, 1732, 1651, 1560, 1375,
1313, 1254, 1117, 1074 ¹ H-NMR (CD ₃ OD): δ 1.23 (3H, t, J = 7 Hz, CH ₂ C H ₃ ) 1.85 (2H, m, C H ₂ ) 1.99 (3H, s, NAc) 2.38 (2H, t, J = 7.5Hz, C H 2 CO) 4.10 (2H, q, J = 7Hz, C H 2 CH 3) 4.44 (1H, d, J 1,2 = 8.5Hz, H- 1) 22: Amorphous powder [α] _D ²⁰ -2.0 ° (c 0.50, MeOH) IR (KBr) ν _max cm ^-1 : 3400, 1734, 1651, 1562, 1375,
1116, 1090 ¹ H-NMR (CD ₃ OD): δ 1.86 (2H, m, C H ₂ ) 1.98 (3H, s, NAc) 2.40 (2H, t, J = 7 Hz, C H ₂ CO) 3.65 (3H , s, OMe) 4.43 (1H, d, J _1,2 = 8.5Hz, H-1)

Reference Example 14 8- (methoxycarbonyl)
Octyl-2,3,4-tri-O-benzyl-α-L-
Fucopyranoside (24) and 8- (methoxycarbonyl) octyl-2,3,4-tri-O-benzyl-β-
L-fucopyranoside (25) [M. Dejter-Juszynski et al., Carbohydr. Res., 18 , 219
(1971)], from 4.00 g (6.85 mmol) of 2,3,4-tri-O-benzyl-1-Op-nitrobenzoyl-β-L-fucopyranose.
1.29 g of 8- (methoxycarbonyl) octanol (12) was added to a solution of 3,4-tri-O-benzyl-α-L-fucopyranosyl bromide (23) in 40 ml of benzene.
(6.86 mmol), 1.73 g of mercuric cyanide
(6.86 mmol) and 3.60 g of powdered anhydrous calcium sulfate were added, and the mixture was stirred and reacted in an argon stream at room temperature for 2 days. After the completion of the reaction, the insoluble material was removed by filtration, 60 ml of ethyl acetate was added to the filtrate, and a saturated aqueous solution of sodium hydrogencarbonate 6 was added.
It was sequentially washed with 0 ml and water 60 ml. After drying over anhydrous magnesium sulfate and concentration under reduced pressure, the syrup obtained was purified by silica gel column chromatography (n-hexane: ethyl acetate = 15: 1) to give the title compounds (24) and (2).
5) at 1.66 g (40.1%) and 522 mg, respectively
(12.6%). 24: syrup ^{_{[α] D 25 - 34.0 °}} (c 1.0, CHCl 3) IR (film) ν max cm -1: 1725 1 H-NMR (CDCl 3): δ 1.10 (3H, d, J = 7.5Hz , H-6), 2.28 (2H, t, J = 7.6Hz, CH ₂ COOMe) 3.65 (3H, s, COOMe) 25: Syrup-like [α] _D ²⁵ + 6.2 ° (c 1.0, CHCl ₃ ) IR (film) ν _max cm ^-1 : 1725 ¹ H-NMR (CDCl ₃ ): δ 1.15 (3H, d, J = 7.5 Hz, H-6) 2.28 (2H, t, J = 7.6 Hz, C H ₂ COOMe ) 3.65 (3H, s, COOMe) 4.32 (1H, d, J = 8.6Hz, H-1)

Reference Example 15 8- (methoxycarbonyl)
Octyl-β-L-fucopyranoside (26) To a solution of 596 mg (0.985 mmol) of compound (25) in 20 ml of acetic acid was added 300 mg of 10% palladium-carbon,
Hydrolysis was carried out for 3 days at room temperature using a Parr apparatus (50 psi). After completion of the reaction, the catalyst was removed by filtration, the filtrate was concentrated to dryness under reduced pressure, and the obtained solid was subjected to silica gel column chromatography (chloroform: methanol = 40:40).
Purified in 1) to give 226 mg of the title compound (26) (68.
6%). 26: Syrup-like [α] _D ²⁵ + 4.5 ° (c 1.0, MeOH) IR (film) ν _max cm ^-1 : 3350, 1740 MS (SIMS): m / z 335 (M + H) ^{+ 1} H-NMR (CD ₃ OD): δ 1.21 (3H, d, J = 7.5Hz, H-6) 2.31 (2H, t, J = 7.6Hz, C H ₂ COOMe) 4.17 (1H, d, J = 8.1Hz, H -1)

Reference Example 16 8- (methoxycarbonyl)
Octyl-2,3,4-tri-O-acetyl-α-D-
Xylopyranoside (28) and 8- (methoxycarbonyl) octyl-2,3,4-tri-O-acetyl-β-
D-xylopyranoside (29) 2,3,4-tri-O-acetyl-prepared by the method of [DH Brauns, J. Am. Chem. Soc., 47 , 1280 (1295)].
α-D-xylopyranosyl bromide (27) 1.20
g (3.54 mmol) in 10 ml of benzene
333 mg of (methoxycarbonyl) octanol (12)
(1.77 mmol), 447 mg of mercuric cyanide
(1.77 mmol) and 1.0 g of powdered anhydrous calcium sulfate were added, and the mixture was stirred at room temperature in an argon stream for 24 hours. After completion of the reaction, the insoluble material was removed by filtration, 20 ml of ethyl acetate was added to the filtrate, and 20 ml of a saturated aqueous sodium hydrogen carbonate solution was added.
Washed sequentially with 20 ml of water. After drying over anhydrous magnesium sulfate and concentrating under reduced pressure, the resulting syrup was subjected to silica gel column chromatography (toluene: ethyl acetate =
8: 1) to give 426 mg (53.9%) and 98 mg (12.4 mg) of the title compounds (28) and (29), respectively.
%)Obtained. 2.2 To a solution of 850 mg (2.51 mmol) of 2,3,4-tri-O-acetyl-α-D-xylopyranosyl bromide (27) in 10 ml of dichloroethane was added 1.61 g (6.28 mmol) of silver triflate. ), 875 mg (7.5 mmol) of 1,1,3,3-tetramethylurea and 8-
(Methoxycarbonyl) octanol (12) 236mg
(1.46 mmol) at room temperature in an argon stream.
Stirred for hours. 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. After drying over anhydrous magnesium sulfate and concentrating under reduced pressure, the obtained syrup was purified by silica gel column chromatography (toluene: ethyl acetate = 8: 1) to give 375 mg (67.0) of the title β-anomer (29). %). 28: Syrup-like [α] _D ²⁵ + 92.4 ° (c 1.0, CHCl ₃ ) IR (film) ν _max cm ^-1 : 2940, 1760, 1230 ¹ H-NMR (CDCl ₃ ): δ 2.03 and 2.06 (6H, 2s, OAc) 2.31 (2H, t, J = 7.6Hz, C H 2 COOMe) 3.67 (3H, s, COOMe) 29: syrup ^{_{[α] D 25 - 19.5 °}} (c 1.0, CHCl 3) IR (film ) ν _max cm ^-1 : 2930,1760,1220 ¹ H-NMR (CDCl ₃ ): δ 2.03 (3H, s, OAc) 2.05 (3H, s, OAc) 2.06 (3H, s, OAc) 2.30 (2H, t, J = 7.4Hz, C H ₂ COOMe) 3.67 (3H, s, COOMe) 4.46 (1H, d, J = 6.8Hz, H-1)

Reference Example 17 8- (methoxycarbonyl)
Octyl-β-D-xylopyranoside (30) To a solution of 101 mg (0.226 mmol) of compound (29) in 2 ml of methanol was added 0.03 ml of 28% sodium methoxide-methanol, and the mixture was stirred at room temperature for 3 hours. The reaction solution was neutralized with Amberlite IR-120B (H ⁺ ), the resin was removed 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 58.3 mg (80.5%) of the title compound (30). 30: syrup ^{_{[α] D 25 - 10.3 °}} (c 1.0, MeOH) IR (film) ν max cm -1: 3450, 2960, 1740, 1290, 1050 1 H-NMR (CD 3 OD): δ 2.31 ( 2H, t, J = 7.3Hz, C H ₂ COOMe) 3.65 (3H, s, COOMe) 4.17 (1H, d, J = 7.6Hz, H-1)

Reference Example 18 6- (methoxycarbonyl)
Hexyl-3,4,6-tri-O-acetyl-2-azido-2-deoxy-α-D-mannopyranoside (33) By the same method as described in Reference Example 24 below,
3,4,6-tri-O-acetyl-2-azido-2-deoxy-α-D-mannopyranosyl bromide (31) 7
30 mg (1.85 mmol) and 271 mg (1.85 mmol) of 6- (methoxycarbonyl) hexanol (32) were condensed with toluene using silver silicate in the presence of powdered molecular sieve 4A. (3
3) 424 mg (49.9%) were obtained. 33: Syrup-like [α] _D ²⁵ + 67.8 ° (c 0.72, CHCl ₃ ) IR (CHCl ₃ ) ν _max cm ^-1 : 2110, 1747, 1436, 1369, 122
8, 1053 ¹ H-NMR (CDCl ₃ ): δ 1.25-1.70 (8H, m, C H ₂ × 4) 2.04 (3H, s, OAc) 2.09 (6H, s, OAc × 2) 2.32 (2H, t _{, J = 6Hz, C H 2} CO) 3.67 (3H, s, COOMe) 3.84-3.87 (1H, m, H5) 4.00 (1H, dd, J 2,3 = 3.5Hz, H-2) 4.08 (1H, _{dd, J 5.6b = 2.5Hz, H} -6b) 4.24 (1H, dd, J 5,6a = 4.5Hz, J 6a, 6b = 10Hz, H-6a) 4.82 (1H, d, J 1,2 = 1.5 Hz, H-1) 5.26-5.44 (2H, m, H-3 and H-4)

Reference Example 19 6- (methoxycarbonyl)
Hexyl-2-azido-2-deoxy-α-D-mannopyranoside (34) In a similar manner to that described in Reference Example 25 below, 420 mg (0.914 mmol) of compound (33) was added to 1M sodium hydroxide in a methanol solution. After de-O-acetylation using a methoxide-methanol solution, the title compound (34) was obtained.
265 mg (87%) were obtained. 34: Syrup-like [α] _D ^22.5 + 75.8 ° (c 0.89, MeOH) IR (film) ν _max cm ^-1 : 3400,2120,1735,1270,1060 ¹ H-NMR (CD ₃ OD): δ 1.28- 1.70 (8H, m, C H ₂ × 4) 2.33 (2H, t, J = 7.5Hz, C H ₂ CO) 3.65 (3H, s, COOMe) 4.77 (1H, s, H-1)

Reference Example 20 6- (methoxycarbonyl)
Hexyl-2-acetamide-2-deoxy-α-D-mannopyranoside (35) In an ethanol solution, 265 mg (0.795 mmol) of compound (34) was prepared in the same manner as described in Reference Example 26 below. After reducing the azide group to an amino group with sodium borohydride in the presence of nickel chloride hexahydrate, N-acetylation was performed with acetic anhydride to obtain 228 mg (82%) of the title compound (35). 35: Syrup-like [α] _D ^23.5 + 38.5 ° (c 0.52, MeOH) IR (film) ν _max cm ^-1 : 3328, 1730, 1650, 1552, 1438,
1377 ¹ H-NMR (CD ₃ OD): δ 1.25-1.70 (8H, m, C H ₂ × 4) 1.99 (3H, s, NAc) 2.33 (2H, t, J = 7.5Hz, C H ₂ CO) 3.65 (3H, s, COOMe) 4.27 (1H, dd, J _2,3 = 4.3Hz, H-2) 4.65 (1H, d, J _1,2 = 1.2Hz, H-1)

Reference Example 21 8- (methoxycarbonyl)
Octyl-3,4,6-tri-O-acetyl-2-azido-2-deoxy-α-D-mannopyranoside (36)
And 8- (methoxycarbonyl) octyl-3,4,6
-Tri-O-acetyl-2-azido-2-deoxy-β
-D-mannopyranoside (37) 3,4,6-tri-O synthesized by the method described in the literature (Carbohydr. Res., 136 , 153 (1985)) in the same manner as described in Reference Example 24 below. -Acetyl-2-azido-2-
Deoxy-α-D-mannopyranosyl bromide (31)
505 mg (1.27 mmol) and 241 mg (1.27 mmol) of 8- (methoxycarbonyl) octanol (12) were condensed in toluene in the presence of powdered molecular sieve 4A using silver silicate to give the title compound ( 3
250 mg (45.2%) of each of 6) and (37),
273 mg (42.3%) were obtained. 36: Syrup-like [α] _D ²³ + 67.0 ° (c 1.3, CHCl ₃ ) IR (CHCl ₃ ) ν _max cm ^-1 : 2028,1745,1739,1370,1053 ¹ H-NMR (CDCl ₃ ): δ 1.2 -1.7 (12H, m, C H ₂ × 6) 2.05 (3H, s, OAc × 2) 2.30 (2H, t, J = 6Hz, C H ₂ CO) 3.67 (3H, s, COOMe) 3.86-3.88 (1H, m, H-5) 4.01 (1H, dd, J _2,3 = 1.4Hz, H-2) 4.09 (1H, dd, J _5.6b = 2.2Hz, H-6b ) 4.25 (1H, dd, J 5,6a = 4.8Hz, J 6a, 6b = 12Hz, H-6a) 4.83 (1H, d, J 1,2 = 1.6Hz, H-1) 5.25-5.42 (2H, m, H-3 and H- 4) 37: syrup ^{_{[α] D 23 - 75.5 °}} (c 0.83, CHCl 3) IR (CHCl 3) ν max cm -1: 2028, 1745, 1439, 1370, 105
3 ¹ H-NMR (CDCl ₃ ): δ 1.2-1.7 (12H, m, C H ₂ × 6) 2.04 (3H, s, OAc) 2.10 (3H, s, OAc) 2.11 (3H, s, OAc) 2.30 (2H, t, J = 7Hz, C H ₂ CO) 3.67 (3H, s, COOMe) 4.13 (1H, dd, J _5.6b = 2.2Hz, H-6b) 4.28 (1H, dd, J _5,6a = 4.8Hz, J _{6a, 6b} = 12Hz, H-6a) 4.66 (1H, d, J _1,2 = 1.5Hz, H-1) 4.98 (1H, dd, J _2,3 = 4Hz, H-4) 5.25 (1H, t, J _3,4 = 10Hz, H-3)

Reference Example 22 8- (methoxycarbonyl)
Octyl-2-azido-2-deoxy-α-D-mannopyranoside (38) According to a method similar to that described in Reference Example 25 below, 1 M was added to 7 ml of a methanol solution of 283 mg (0.564 mmol) of compound (36). The mixture was de-O-acetylated by adding 0.1 ml of a sodium methoxide-methanol solution to obtain 210 mg (quantitative) of the title compound (38). 38: Syrup-like [α] _D ²⁴ + 68.6 ° (c 0.61, MeOH) IR (CHCl ₃ ) ν _max cm ^-1 : 3600, 2028, 1730 ¹ H-NMR (CD ₃ OD): δ 1.2-1.7 (12H , m, C H ₂ × 6) 2.11 (3H, s, OAc) 2.32 (2H, t, J = 7Hz, C H ₂ CO) 3.65 (3H, s, COOMe) 4.77 (1H, d, J _1,2 = 1.5Hz, H-1)

Reference Example 23 8- (methoxycarbonyl)
Octyl-2-acetamide-2-deoxy-α-D-mannopyranoside (39) A solution of 210 mg (0.559 mmol) of compound (38) in 5 ml of ethanol was prepared in the same manner as described in Reference Example 26 below. After adding 0.1 ml of a 0.6 M nickel chloride hexahydrate-ethanol solution, the azide group was reduced to an amino group with a solution of 63 mg (1.677 mmol) of sodium borohydride in 5 ml of ethanol. Then, N-acetylation was performed with 0.2 ml of acetic anhydride to give the title compound (39) 1.
47 mg (67.4%) were obtained. 39: Syrup-like [α] _D ²⁵ + 37.1 ° (c 0.42, MeOH) IR (CHCl ₃ ) ν _max cm ^-1 : 1732, 1655, 1439, 1375, 113
0, 1070, 1027 ¹ H-NMR (CD ₃ OD): δ 1.25-1.75 (12H, m, C H ₂ × 6) 2.00 (3H, s, NAc) 2.32 (2H, t, J = 7.5Hz, C H ₂ CO) 3.65 (3H, s, COOMe) 4.28 (1H, dd, J _2,3 = 4.8Hz, H-2) 4.66 (1H, d, J _1,2 = 1.6Hz, H-1)

Reference Example 24 2- [2- (3-methoxycarbonylpropyloxy) ethoxy] ethyl-4-O-
(2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl) -3,6-di-O-acetyl-2-
Azido-2-deoxy-β-D-glucopyranoside (4
1) Literature description (H. Paulsen et al., Liebigs Ann. Chem., 1
121 (1982)).
-Tetra-O-acetyl-β-D-galactopyranosyl) -3,6-di-O-acetyl-2-azido-2-deoxy-β-D-glucopyranosyl bromide (40) 6
00 mg (0.879 mmol), methyl 4- [2- (2
-Hydroxyethoxy) ethoxy] -1-butanoate (6) 900 mg of silver silicate was added to a solution of 182 mg (0.879 mmol) of powdery molecular sieve 4A and 1.5 g of toluene in 25 ml of toluene in a nitrogen stream under ice-cooling. After the reaction mixture was stirred at the same temperature for 3 hours, the cooling bath was removed, and the mixture was further stirred at room temperature for 16 hours. After completion of the reaction, the reaction mixture was filtered using celite, and the filtrate was concentrated to dryness under reduced pressure.
The obtained residue (700 mg) was crystallized from ether to give the title compound (41) (455 mg, 64%). 41: mp 109-111 ° C [α] _D ²² + 2.7 ° (c 0.45, CHCl ₃ ) IR (CHCl ₃ ) ν _max cm ^-1 : 2116, 1751, 1369, 1064 ¹ H-NMR (CDCl ₃ ): δ 1.93 (3H, s, OAc) 2.00 (3H, s, OAc) 2.04 (3H, s, OAc) 2.08 (3H, s, OAc) 2.09 (3H, s, OAc) 2.12 (3H, s, OAc) 2.38 ( 2H, t, J = 7.5Hz, C H 2 CO) 3.64 (3H, s, COOMe) 3.75 (1H, m, C H 2 O) 3.84 (1H, t, J 5'.6'a = 6Hz, H -5 ') 3.95 (1H, m, C H ₂ O) 4.02-4.08 (2H, m) 4.13 (1H, dd, J _{5', 6'a} = 6Hz, J _{6'a, 6'b} = 12.5Hz , H-
6'a) 4.41 (1H, d, J _{1 ', 2'} = 8Hz, H-1) 4.44 (1H, d, J _1,2 = 8Hz, H-1) 4.92 (1H, dd, H-3 ' ) 4.94 (1H, dd, J _2,3 = 9.5Hz, J _3,4 = 10.2Hz, H-3) 5.04 (1H, dd, J _{1 ', 2'} = 8Hz, J _{2 ', 3'} = 10.5 Hz, H-2 ') 5.31 (1H, dd, J _{3', 4 '} = 3Hz, J _{4', 5 '} = 1.0Hz, H-4')

Reference Example 25 2- [2- (3-methoxycarbonylpropyloxy) ethoxy] ethyl-4-O-
(Β-D-galactopyranosyl) -2-azido-2-deoxy-β-D-glucopyranoside (42) 1 M sodium methoxide-methanol was added to a solution of 450 mg (0.557 mmol) of compound (41) in 20 ml of methanol. 0.23 ml of the solution was added and left at room temperature for 20 hours. Add ion exchange resin Amberlite IR-12 to the reaction solution.
After adding 0B (H ⁺ type) for neutralization, the resin was removed by filtration and washed with methanol. The combined filtrate and washings were concentrated to dryness under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform: methanol = 5: 1) to give 280 mg (90.6%) of the title compound (42).
I got 42: amorphous powder ^{_{[α] D 23 - 3 °}} (c 0.36, MeOH) IR (KBr) ν max cm -1: 3400, 2120, 1738, 1439, 1367,
1317, 1255, 1169, 1139, 1078 ¹ H-NMR (CD ₃ OD): δ 1.85 (2H, m, C H ₂ ) 2.40 (2H, t, J = 7.5 Hz, C H ₂ CO) 3.65 (3H, s, COOMe) 4.35 (1H, d, J _{1 ', 2'} = 7.5Hz, H-1 ') 4.43 (1H, d, J _1,2 = 10Hz, H-1)

Reference Example 26 2- [2- (3-methoxycarbonylpropyloxy) ethoxy] ethyl-4-O-
(Β-D-galactopyranosyl) -2-acetamide-2
-Deoxy-β-D-glucopyranoside (43) 380 mg of nickel chloride hexahydrate was added to a solution of 270 mg (0.486 mmol) of compound (42) in 5 ml of ethanol.
(1.6 mmol) dissolved in 10 ml of ethanol was added to 0.1 ml of sodium borohydride (55 mg).
A solution of (1.458 mmol) in 1.1 ml of ethanol was added with stirring. After stirring at room temperature for 30 minutes, the reaction solution was neutralized by adding acetic acid. Then, 0.5 ml of acetic anhydride was added, and the mixture was left at room temperature for 16 hours. Thereafter, the reaction mixture was concentrated to dryness under reduced pressure, and the obtained residue was purified by silica gel column chromatography [chloroform: methanol: water = 7: 3: 1 (lower layer)] to give the title compound (4
3) 171 mg (62%) were obtained. 43: amorphous powder ^{_{[α] D 20 - 14.7 °}} (c 0.54, MeOH) IR (KBr) ν max cm -1: 3422, 1735, 1653, 1558, 1438,
1377, 1302 ¹ H-NMR (CD ₃ OD): δ 1.85 (2H, m, C H ₂ ) 1.96 (3 H, s, NAc) 2.40 (2 H, t, J = 7.5 Hz, C H ₂ CO) 3.65 ( 3H, s, COOMe) 4.37 (1H, d, J _1,2 = 8Hz, H-1) 4.48 (1H, d, J _{1 ', 2'} = 8Hz, H-1 ')

Reference Example 27 8- (methoxycarbonyl)
Octyl-2-acetamide-3,4-di-O-acetyl-2-deoxy-6-O-trityl-β-D-glucopyranoside (45) [RULemieux et al., J. Am. Chem. Soc., 97 , 4076. (1975)], 895 mg (2.29 mmol) of 8- (methoxycarbonyl) octyl-2-acetamido-2-deoxy-β-D-glucopyranoside (44) was prepared.
After dissolving in 20 ml of pyridine, the mixture was concentrated under reduced pressure to about 5 ml. 715 mg (2.56 mmol) of trityl chloride was added, and the mixture was stirred overnight at room temperature. After completion of the reaction, the reaction solution was added with water 0.1.
After adding 1 ml and stirring for 10 minutes, the mixture was concentrated under reduced pressure.
The obtained syrup was dissolved in chloroform (30 ml) and washed sequentially with saturated aqueous sodium hydrogen carbonate solution (30 ml) and water (30 ml). After drying over anhydrous magnesium sulfate, 5 ml of pyridine and 5 ml of acetic anhydride were added to the syrup obtained by concentration under reduced pressure, and the mixture was stirred at room temperature for 4 hours. 2 ml of methanol was added to the reaction solution under ice cooling, and the mixture was stirred for 20 minutes and concentrated under reduced pressure.
The obtained syrup was purified by silica gel column chromatography (chloroform: methanol = 120: 1) to obtain 1.12 g (68.3%) of the title compound (45). 45: mp 159-160 ° C. [α] _D ²⁵ + 31.8 ° (c 1.0, CHCl ₃ ) IR (KBr) ν _max cm ⁻¹ : 1750, 1240 ¹ H-NMR (CDCl ₃ ): δ 1.71 (3H, s, NAc) 1.94 (3H, s, OAc) 2.00 (3H, s, OAc) 2.27 (2H, t, J = 7.3Hz, C H 2 COOMe) 3.65 (3H, s, COOMe) 4.61 (1H, d, J = 8.3Hz, H-1) 5.51 (1H, d, J = 8.8Hz, NH)

Reference Example 28 8- (methoxycarbonyl)
Octyl-2-acetamido-3,4-di-O-acetyl-2-deoxy-β-D-glucopyranoside (46) After dissolving 1.00 g (1.39 mmol) of compound (45) in 5 ml of acetic acid, 25 1 ml of an acetic acid solution containing 5% hydrogen bromide was added thereto, followed by stirring at 15 ° C. for 1.5 minutes. After the resulting precipitate was collected by filtration, the filtrate was added to 30 ml of cold water and added with 30 ml of chloroform.
Extracted times. Thereafter, it was washed with 30 ml of water and dried over anhydrous magnesium sulfate. The syrup obtained by concentration under reduced pressure was purified by silica gel column chromatography (chloroform: methanol = 40: 1) to obtain 493 mg (74.6%) of the title compound (46). 46: syrup ^{_{[α] D 25 - 11.2 °}} (c 1.0, CHCl 3) IR (KBr) ν max cm -1: 1740, 1660, 1250 1 H-NMR (CDCl 3): δ 1.94,2.04 and 2.05 ( 9H, 3s, NAc and OAc × 2) 2.30 (2H, t, J = 7.6Hz, C H ₂ COOMe) 3.67 (3H, s, COOMe) 4.69 (1H, d, J = 8.3Hz, H-1) 5.54 (1H, d, J = 8.6Hz, H-1)

Reference Example 29 8- (methoxycarbonyl)
Octyl-2-acetamido-3,4-di-O-acetyl-2-deoxy-6-O- (2,3,4-tri-O-
Acetyl-β-L-fucopyranosyl) -β-D-glucopyranoside (48) To a solution of 120 mg (0.252 mmol) of compound (46) in 8 ml: 8 ml of benzene: nitromethane was added 1,2,2
3,4-tetra-O-acetyl-L-fucopyranose 1
From 26 mg (0.379 mmol) [HMFlowers et al., Ca
rbohydr. Res., 4,189 (1967)].
A solution of 3,4-tri-O-acetyl-α-L-fucopyranosyl bromide (47) in 2 ml of ethylene dichloride and 63.7 mg (0.252 mmol) of mercuric cyanide were added, and the mixture was stirred at room temperature in an argon stream. For 3 days. 20 ml of chloroform was added to the reaction solution, which was washed successively with 20 ml of a saturated aqueous solution of sodium hydrogen carbonate and 20 ml of water. After drying over anhydrous magnesium sulfate, the solution was concentrated under reduced pressure, and the resulting syrup was purified by silica gel column chromatography (chloroform: methanol = 200: 1) to give compound (4).
8) 86.4 mg (30.5%) were obtained. Further, 45.3 mg (37.8%) of the compound (46) was recovered. 2. To a solution of 120 mg (0.252 mmol) of compound (46) in 4 ml of methylene chloride, 227 mg of silver triflate (0.2 mg) was added.
882 mmol), 176 mg (1.51 mmol) of 1,1,3,3-tetramethylurea and 176 mg of 1,2,3,4-tetra-O-acetyl-L-fucopyranose
(0.379 mmol) to [HMFlowers et al., Carbohyd
r. Res., 4 , 189 (1967)].
A solution of 4-tri-O-acetyl-α-L-fucopyranosyl bromide (47) in 2 ml of ethylene dichloride was added, and the mixture was stirred at room temperature in an argon stream for 24 hours. After completion of the reaction, 20 ml of chloroform was added to the reaction solution, which was washed successively with 20 ml of a saturated aqueous solution of sodium hydrogen carbonate and 20 ml of water. After drying over anhydrous magnesium sulfate and concentration under reduced pressure, the syrup obtained was subjected to silica gel column chromatography (chloroform:
Purification with methanol = 200: 1) gave the title compound (4
8) 136 mg (72.2%) were obtained. 48: Syrup-like [α] _D ²⁵ + 4.5 ° (c 1.0, CHCl ₃ ) IR (KBr) ν _max cm ^-1 : 2940, 1750 ¹ H-NMR (CDCl ₃ ): δ 1.19 (3H, d, J = 6.4Hz, H-6 ') 1.92 (3H, s) 1.96 (3H, s) 1.99 (3H, s) 2.00 (3H, s) 2.10 (3H, s) 2.15 (3H, s) 2.29 (2H, t, J = 7.6Hz, C H ₂ COOMe) 3.65 (3H, s, COOMe) 4.5-4.6 (2H, m, H-1 and H-1 ') 5.49 (1H, d, J = 8.8Hz, NH)

Reference Example 30 8- (methoxycarbonyl)
Octyl-2-acetamido-2-deoxy-6-O-
(Β-L-fucopyranosyl) -β-D-glucopyranoside (49) To a methanol solution of 125 mg (0.167 mmol) of compound (48) was added 0.04 ml of 28% sodium methoxide-methanol, and the mixture was allowed to stand at room temperature overnight. Stirred. The reaction solution was neutralized with Amberlite IR-120B (H ⁺ ), the resin was removed by filtration, and the filtrate was concentrated under reduced pressure. The obtained syrup was purified by silica gel column chromatography (chloroform: methanol: water = 16: 4: 1),
68.9 mg (76.7%) of the title compound (49) were obtained. 49: syrup ^{_{[α] D 25 - 16.7 °}} (c 1.0, MeOH) IR (film) ν max cm -1: 3380, 1750, 1080 1 H-NMR (CD 3 OD): δ 1.67 (3H, d, J = 6.6Hz, H-6 ' ) 1.96 (3H, s, NAc) 2.30 (2H, t, J = 7.6Hz, C H 2 COOMe) 3.64 (3H, s, COOMe) 4.24 (1H, d, J = 7.3Hz, H-1 ') 4.40 (1H, d, J = 8.3Hz, H-1)

Reference Example 31 8- (methoxycarbonyl)
Octyl-2,4-di-O-benzyl-α-D-mannopyranoside (53) Reference (T. OGAWA et al., Tetrahedron, 37, 2787 (1981))
3,6-di-O-acetyl-2,4- synthesized by the method of
16 g (36.0 mmol) of di-O-benzyl-α-D-mannopyranosyl chloride (51) and 7.11 g of 8- (methoxycarbonyl) octanol (12) (3
(7.77 mmol) in 270 ml of toluene was cooled to -18 ° C under a stream of nitrogen. To this solution, 17.5 g of powdered molecular sieve 4A was added and stirred for 30 minutes. Then, 16 g of silver silicate was added and the mixture was stirred at the same temperature for 24 hours. After the reaction mixture was filtered using celite, the filtrate was concentrated to dryness under reduced pressure. The resulting syrupy residue was purified by silica gel column chromatography (n-hexane: ethyl acetate = 3: 1) to obtain a crude condensation product 27.5.
g. 27.5 g of the condensation product was added to 200 ml of methanol.
Then, 1 ml of a 1M sodium methoxide-methanol solution was added, and the mixture was allowed to stand at room temperature for 16 hours. The reaction solution was ion-exchange resin Amberlite IR-120B (H ⁺
) Was added to neutralize the mixture, and the resin was removed by filtration and washed with methanol. The filtrate and washings were combined and concentrated to dryness under reduced pressure. The obtained residue was purified by silica gel column chromatography to give 7.19 g (30) of the title compound (53).
%) And 13.6 g (57%) of a mixture of compound (53) and its β-anomer. 53: Syrup-like [α] _D ²⁴ + 6.7 ° (c 0.84, CHCl ₃ ) IR (CHCl ₃ ) ν _max cm ^-1 : 3560, 1731, 1498, 1455, 143
8, 1075 ¹ H-NMR (CDCl ₃ ): δ 1.20-1.70 (12H, m, C H ₂ × 6) 2.28 (2H, t, J = 7.5 Hz, C H ₂ CO) 3.64 (3H, s, COOMe ) 4.59 (1H, d, J = 12Hz, C H ₂ Ph) 4.64 (1H, d, J = 11Hz, C H ₂ Ph) 4.71 (1H, d, J = 12Hz, C H ₂ Ph) 4.82 (1H, s, H-1) 4.89 (1H, d, J = 11Hz, C H ₂ Ph) 7.20-7.40 (10H, m, aromatic H)

Reference Example 32 8- (methoxycarbonyl)
Octyl-6-O- (3,4,6-tri-O-benzyl-α-D-mannopyranosyl) -2,4-di-O-benzyl-α-D-mannopyranoside (55) and 8- (methoxycarbonyl) ) Octyl-3,6-di-O- (3,
4,6-Tri-O-benzyl-α-D-mannopyranosyl) -2,4-di-O-benzyl-α-D-mannopyranoside (56) described in the literature (T. Ogawa et al., Tetrahedron, 37, 2779 (1981). ))
3.5 g (6.84 mmol) of 2-O-acetyl-3,4,6-tri-O-benzyl-α-D-mannopyranosyl chloride (54), compound (5)
3) 1.52 g (2.852 mmol) and 1,1,
3.638 ml of 3,3-tetra-N-methylurea (30.
(81 mmol) in 70 ml of dichloroethane at −18 ° C. under a nitrogen stream with stirring at 1.18 ° C.
5 g (7.53 mmol) were added. The cooling bath was removed from the reaction mixture, and the mixture was stirred at room temperature for 18 hours. After the reaction mixture was filtered using celite, the filtrate was washed successively with an aqueous solution of sodium hydrogen carbonate and water. After drying over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure. The obtained residue is subjected to row bar column C (n-hexane: ethyl acetate: = 2: 1).
To give 4.21 g of a crude condensation product. After dissolving 3.17 g of the condensation product in 100 ml of methanol, 1M sodium methoxide-methanol solution 8
0.5 ml was added and left at room temperature for 48 hours. The reaction solution was ion-exchange resin Amberlite IR-120B (H ⁺
) Was added to neutralize the mixture, and the resin was removed by filtration and washed with methanol. The filtrate and washings were combined and concentrated to dryness under reduced pressure. The obtained residue (3.01 g) was placed in a row bar column C (n-
Purification with hexane: ethyl acetate = 2: 1) gave 696 mg (20%) of the title compound (55) and 1.47 g of (56).
(27%). 55: Syrup-like [α] _D ^22.5 + 49.6 ° (c 1.02, CHCl ₃ ) IR (CHCl ₃ ) ν _max cm ^-1 : 1731, 1602, 1455, 1099, 107
6 ¹ H-NMR (CDCl ₃ ): δ 1.20-1.65 (12H, m, C H ₂ × 6) 2.27 (2H, t, J = 8 Hz, C H ₂ CO) 3.64 (3H, s, COOMe) 3.69 ( 1H, dd, J _2,3 = 3.5Hz, H-2) 4.08 (1H, d, J _{2 ', 3'} = 2.0Hz, H-2 ') 4.84 (1H, d, J _1,2 = 1.0Hz , H-1) 5.01 (1H, d, J _{1 ', 2'} = 1.5Hz, H-1 ') 7.10-7.40 (25H, m, aromatic H) 56: Syrup-like [α] _D ^22.5 + 45.5 ° ( c 1.04, CHCl ₃ ) IR (CHCl ₃ ) ν _max cm ^-1 : 1730, 1602, 1454, 1101, 107
4 ¹ H-NMR (CDCl ₃ ): δ 1.20-1.70 (12H, m, C H ₂ × 6) 2.26 (2H, t, J = 7.5 Hz, C H ₂ CO) 4.74 (1H, d, J _{1, 2} = 1.5Hz, H-1) 5.01 (1H, d, J _{1 '', 2 ''} = 1.0Hz, H-1 '') 5.21 (1H, s, H-1 ') 7.10-7.40 (40H, m, aromatic H)

Reference Example 33 8- (methoxycarbonyl)
Octyl-6-O- (α-D-mannopyranosyl) -α
-D-Mannopyranoside (57) A solution of 690 mg (0.737 mmol) of compound (55) in 30 ml of methanol was mixed with 180 mg of 10% palladium-carbon.
Was added and hydrogenolysis was performed at room temperature for 48 hours. After completion of the reaction, the catalyst was removed by filtration, and the filtrate was concentrated to dryness under reduced pressure. The obtained residue was purified by silica gel column chromatography [chloroform: methanol: water = 7: 3: 1 (lower layer)] to obtain 146 mg of the title compound (57). 57: Amorphous powder [α] _D ²³ + 64.7 ° (c 0.59, MeOH) IR (KBr) ν _max cm ^-1 : 3400, 1740, 1437, 1132, 1099,
1064 ¹ H-NMR (CD ₃ OD): δ 1.25-1.60 (12H, m, C H ₂ × 6) 2.31 (2H, t, J = 7.5 Hz, C H ₂ CO) 3.64 (3H, s, COOMe) 4.69 (1H, d, J _1,2 = 2.0Hz, H-1) 4.81 (1H, d, J _{1 ', 2'} = 1.5Hz, H-1 ')

Reference Example 34 8- (methoxycarbonyl)
Octyl-3,6-di-O- (α-D-mannopyranosyl) -α-D-mannopyranoside (58) A mixture of 300 mg (0.214 mmol) of compound (56) in 20 ml of methanol, 7 ml of water and 7 ml of acetic acid was added. % Palladium on carbon was added and hydrogenolysis was performed at room temperature for 16 hours. After removing the catalyst by filtration, the filtrate was concentrated to dryness under reduced pressure to obtain 144 mg (quantitative) of the title compound (58). 58: Amorphous powder [α] _D ²³ + 56.4 ° (c 0.5, MeOH) IR (KBr) ν _max cm ^-1 : 3423, 1731, 1438, 1247, 1050 ¹ H-NMR (CD ₃ OD): δ 1.25 -1.70 (12H, m, C H ₂ × 6) 2.30 (2H, t, J = 7.5Hz, C H ₂ CO) 3.63 (3H, s, COOMe) 4.00 (1H, dd, J _{1 '', 2 ''} = 1.5Hz, J _2'',3'' = 3Hz, H-
2 '') 4.67 (1H, d, J _1,2 = 1.5Hz, H-1) 3.95 (1H, dd, J _2,3 = 3.2Hz, H-2) 4.80 (1H, d, J _{1 ', 2 '} = 1.5Hz, H-1') 5.04 (1H, d, J _{1 '', 2 ''} = 1.5Hz, H-1 '') 7.10-7.40 (40H, m, aromatic H)

Reference Example 35 Poly-N (ε)-[3- (4
-Hydroxyphenyl) propanoyl] -L-lysine hydrobromide (64) Poly-L-lysine hydrobromide (average molecular weight 11,70
75 mg (6.14 micromoles)
08M sodium tetraborate (Na ₂ B ₄ O ₇ ) and 0.1M sodium tetraborate.
Dissolve in 10 ml of 35M potassium bicarbonate (KHCO ₃ ) and at 4-5 ° C. 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (6
2) 8.4 mg (31.9 micromol) of dry N, N-
After dropping 0.2 ml of dimethylformamide solution, 4 ° C
And stirred for 18 hours in the same chamber. The pH of the reaction solution was adjusted to 1.0 by adding concentrated hydrobromic acid, and the
The solution was dialyzed against pure water using a dialysis membrane of pore 3 (molecular weight cut-off = about 3,500), and the obtained dialysate was lyophilized to give 41.6 mg of the complex (64) as a colorless solid. Obtained. ^{_{64: [α] D 21 -}} 47.25 ° (c 0.51, H 2 O) IR (KBr) ν max cm -1: 3400, 3300, 1650, 1540

Reference Example 36 Poly-N (ε)-[3- (4
-Hydroxyphenyl) propanoyl] -L-lysine hydrobromide (65) Poly-L-lysine hydrobromide (average molecular weight 21,70
0) (60) 150 mg (6.9 micromoles)
08M sodium tetraborate (Na ₂ B ₄ O ₇ ) and 0.1M sodium tetraborate.
Dissolve in 10 ml of 35M potassium bicarbonate (KHCO ₃ ) and at 4-5 ° C. 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (6
2) A solution of 10 mg (38 μmol) of 0.3 ml of dry N, N-dimethylformamide was added dropwise, and the mixture was stirred and reacted in a chamber at 4 ° C. for 18 hours. The pH of the reaction solution was adjusted to 0.85 by adding concentrated hydrobromic acid, and the solution was dialyzed against pure water using a dialysis membrane of Spectra / Pore 3 (molecular weight cutoff = about 3,500). The dialyzed solution was freeze-dried to obtain 143.8 mg of the complex (65) as a colorless solid. ^{_{65: [α] D 20 -}} 63.1 ° (c 0.52, H 2 O) IR (KBr) ν max cm -1: 3270, 1625, 1520

Reference Example 37 Poly-N (ε)-[3- (4
-Hydroxyphenyl) propanoyl] -L-lysine hydrobromide (66) poly-L-lysine hydrobromide (average molecular weight 59,00
0) (61) 150 mg (2.54 micromol)
Dissolve in 0.08 M sodium tetraborate (Na ₂ B ₄ O ₇ ) and 10 ml of 0.35 M potassium hydrogen carbonate (KHCO ₃ ) and at 4-5 ° C. N-hydroxy 3- (4-hydroxyphenyl) propionate Succinimide ester (6
2) A solution of 6 mg (22.8 μmol) of 0.3 ml of dry N, N-dimethylformamide was added dropwise, and the mixture was stirred and reacted in a chamber at 4 ° C. for 18 hours. The reaction solution was adjusted to pH 1.4 by adding concentrated hydrobromic acid, and dialyzed against pure water using a dialysis membrane of Spectra / Pore 3 (molecular weight cutoff = about 3,500). The dialysate was freeze-dried to obtain 151.2 mg of the complex (66) as a colorless solid. ^{_{66: [α] D 20 -}} 64.56 ° (c 0.57, H 2 O) IR (KBr) ν max cm -1: 3450, 3300, 1630, 1530

Reference Example 38 Poly-N (ε)-[3- (4
-Hydroxyphenyl) propanoyl] -N (ε)-
[(2-Methoxyethoxy) acetyl] -L-lysine (67) 360 mg (2.6 mg) of 2-methoxyethoxyacetic acid was added to the reaction solution of compound (64) obtained by the method described in Reference Example 35.
3 mmol) and N-hydroxysuccinimide 320
mg (2.78 mmol) in a solution of 2 ml of dry N, N-dimethylformamide was added under ice-cooling to a solution of 530 mg (2.78 mmol) of 1,3-dicyclohexylcarbodiimide in 3 ml of dry tetrahydrofuran. A solution of 2-methoxyethoxyacetic acid N-hydroxysuccinimide active ester (63) prepared by stirring under the above conditions was added. Thereafter, the mixture was stirred and reacted in a chamber at 4 ° C. for 18 hours to precipitate insoluble substances by a centrifugal separator.
Dialysis was performed using the dialysis membrane of (00). The obtained dialysis solution was taken out and freeze-dried to obtain 40.5 mg of the complex (67) as a colorless solid. Fluorescamine method [P. Bohlen,
S. Stein, W. Dairman and S. Udenfriend, Arch. Bioche
m. Biophys., 155 , 213-220 (1973)], the number of remaining amino groups was calculated to be 4. ^{_{67: [α] D 20 -}} 25.1 (c 0.52, H 2 O) IR (KBr) ν max cm -1: 3450, 3300, 1630, 1530

Reference Example 39 Poly-N (ε)-[8- (β
-D-mannopyranosyloxy) nonanoyl] -N
(Ε)-[3- (4-Hydroxyphenyl) propanoyl) -L-lysine hydrobromide (68a) 8- (methoxycarbonyl) octyl-β-D-mannopyranoside (14) was dissolved in methanol and excess 8- (hydrazinocarbonyl) octyl-β-D-mannopyranoside 67. obtained by adding hydrazine and reacting overnight.
3 mg (0.192 mmol) was dissolved in 1 ml of dry N, N-dimethylformamide, and 0.15 ml of a 4N hydrochloric acid-dioxane solution was added under a nitrogen stream, followed by cooling to -20 ° C. To this solution, a solution of 28 mg of t-butyl nitrite in 0.2 ml of dry N, N-dimethylformamide was added, and the mixture was stirred for 30 minutes to obtain an acid azide. Hydrochloride (average molecular weight 11,70
0) (59) 75 mg (6.41 micromoles).
08M sodium tetraborate (Na ₂ B ₄ O ₇ ), 0.35
M Potassium hydrogen carbonate (KHCO ₃ ) was added to a solution dissolved in 10 ml. Next, N-hydroxysuccinimide ester of 3- (4-hydroxyphenyl) propionic acid (6
2) A solution of 8.4 mg (32 μmol) in 0.3 ml of dry N, N-dimethylformamide was added dropwise, and the mixture was stirred and reacted in a chamber at 4 ° C. for 18 hours. The pH of the reaction solution was adjusted to 0.85 by adding concentrated hydrobromic acid, and dialyzed against pure water for 30 hours using a dialysis membrane of Spectra / pore 3 (molecular weight cutoff = about 3,500). The obtained dialysis solution is taken out and lyophilized to give 8 as a colorless solid.
6 mg of the complex (68a) was obtained. Sugar content is phenol-
When colorimetrically determined by the sulfuric acid method, 23 mol of 8- (β-D-mannopyranosyloxy) per mol of poly-L-lysine was determined.
It was found that a nonanoyl group was introduced. Also 3-
When the (4-hydroxyphenyl) propanoyl group content was quantified by absorbance (276 nm), it was found that 5 mol was introduced. Table 1 shows the sugar contents and the number of 3- (4-hydroxyphenyl) propanoyl groups introduced. The physical constants of the compounds are shown in Table 2.

[0081]

[Table 1]

[0082]

[Table 2]

Reference Example 40 Poly-N (ε)-[8- (β
-D-galactopyranosyloxy) nonanoyl] -N
(Ε)-[3- (4-hydroxyphenyl) propanoyl] -L-lysine hydrobromide (68b) 8- (methoxycarbonyl) octyl-β-D-galactopyranoside (15) [DRBundle, DABaker and R.
U. Lemieux, US Patent 4,137,401 (1979) and USP 4,23
8,473 (1980)] was treated with hydrazine monohydrate to obtain 8- (hydrazinocarbonyl) octyl-β-D-galactopyranoside 3.
5 mg (0.0895 mmol) was dissolved in 1 ml of dry N, N-dimethylformamide, 0.15 ml of a 4N hydrochloric acid-dioxane solution was added under a nitrogen stream, and the mixture was cooled to -20 ° C.
To this solution was added a solution of t-butyl nitrite (18 mg) in dry N, N-dimethylformamide (0.1 ml), and the mixture was stirred for 30 minutes to obtain an acid azide. Hydrochloride (average molecular weight 1
1,700) (59) 30 mg (2.56 micromol)
With 0.08 M sodium tetraborate (Na ₂ B ₄ O ₇ )
The solution was added dropwise to a solution dissolved in 6 ml of 0.35 M potassium hydrogen carbonate (KHCO ₃ ). Next, a solution of 6.7 mg (25.4 μmol) of 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) in 0.1 ml of dry N, N-dimethylformamide was added dropwise. The reaction was carried out by stirring in a chamber at a temperature of 18 ° C. for 18 hours. The same post-treatment as in Reference Example 39 was performed to obtain 46.7 mg of a complex (68b) as a colorless solid. Table 1 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 41 Poly-N (ε)-[8- (2
-Acetamido-2-deoxy-β-D-galactopyracylnooxy) nonanoyl] -N (ε)-[3- (4-hydroxyphenyl) propanoyl] -L-lysine hydrobromide (68c) 8- ( Methoxycarbonyl) octyl-2-deoxy-
2-acetamide-β-D-galactopyranoside (16)
[RM Ratcliffe, DA Baker and RU Lemieux, Carb
ohydr. Res., 93 (1981) 35-41.] was treated with hydrazine monohydrate to give 8- (hydrazinocarbonyl) octyl-2-deoxy-2-acetamide. -Β-D-galactopyranoside 6
2.6 mg (0.16 mmol) was converted to acid azide according to the reference example described above and bound to 75 mg (6.41 micromol) of poly-L-lysine hydrobromide (average molecular weight 11,700) (59). I let it. Thereafter, a solution of 8.4 mg (32 micromol) of 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) in 0.1 ml of dry N, N-dimethylformamide was added dropwise, and the mixture was heated at 4 ° C. The reaction was performed by stirring in a chamber for 18 hours. The same post-treatment as in Reference Example 39 was carried out to obtain 63.4 mg of a complex (68c) as a colorless solid. To determine the sugar content, a portion was heated and stirred with 4N hydrochloric acid at 90-100 ° C for 6 hours. The resulting hydrolysis product is Elson-
When colorimetrically determined by the Morgan method, 8- (2-acetamide-
The 2-deoxy-β-D-galactopyranosyloxy) nonanoyl group content is 18 per mole of poly-L-lysine.
Mole. Table 1 shows the number of introduced 3- (4-hydroxyphenyl) propanoyl groups and the sugar content. The physical constants of the compounds are shown in Table 2.

Reference Example 42 Poly-N (ε)-{4- [2
-(2- (2-acetamido-2-deoxy-β-D-galactopyranosyl) ethoxy) ethoxy] butanoyl}
-N (ε)-[3- (4-hydroxyphenyl) propanoyl] -L-lysine hydrobromide (68d) 2- [2- (3-methoxycarbonylpropyloxy)
Ethoxy] ethyl-2-acetamido-2-deoxy-β
2- [2- (3-Hydrazinobonyloxy) ethoxy] ethyl-2-acetamide-2-deoxy-β-D obtained by reacting -D-galactopyranoside (22) with hydrazine monohydrate. -Galactopyranoside,
3- (4-Hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) was prepared in Reference Example 39.
The compound was coupled with poly-L-lysine hydrobromide (average molecular weight 11,700) (59) by the method described in (1) to obtain a complex (68d). Table 1 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups.
The physical constants of the compounds are shown in Table 2.

Reference Example 43 Poly-N (ε)-[8- (β
-L-fucopyranosyloxy) nonanoyl] -N (ε)
-[3- (4-hydroxyphenyl) propanoyl]-
L-Lysine hydrobromide (68e) 8- (Hydrazinocarbonyl) octyl-β-L obtained by treating 8- (methoxycarbonyl) octyl-β-L-fucopyranoside (26) with hydrazine monohydrate
-Fucopyranoside (50 mg, 0.149 mmol) was dissolved in dry N, N-dimethylformamide (1 ml), and thereto was added a 4N hydrochloric acid-dioxane solution (0.15 ml) under a nitrogen stream, and -2.
Cooled to 0 ° C. To this solution was added a solution of 28 mg of t-butyl nitrite in 0.2 ml of dry N, N-dimethylformamide, and the mixture was stirred for 30 minutes to obtain an acid azide. 75 mg (6.41 micromoles) of the hydrochloride (molecular weight 11,700) (59) was added to 0.08 M sodium tetraborate (Na ₂ B).
₄ O ₇ ) and 0.35 M potassium bicarbonate (KHCO
₃ ) The solution was added dropwise to a solution dissolved in 10 ml, and the solution was added to Reference Example 39.
And treated as a colorless solid in 5%.
7.9 mg of the complex (68e) were obtained. Sugar content and 3-
Table 1 shows the number of introduced (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 44 Poly-N (ε)-[8- (2
-Acetamide-2-deoxy-α-D-mannopyranosyloxy) nonanoyl] -N (ε)-[3- (4-hydroxyphenyl) propanoyl] -L-lysine hydrobromide (68 g) 8 8- (Hydrazinocarbonyl) octyl-2-acetamido-2-deoxy obtained by reacting-(methoxycarbonyl) octyl-2-acetamido-2-deoxy-α-D-mannopyranoside (39) with hydrazine monohydrate -Α-D-mannopyranoside and 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) were prepared by the method described in Reference Example 39 using poly-L-lysine hydrobromide (average). Molecular weight 11,
700) (59) to give the complex (68 g). Table 1 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 45 Poly-N (ε)-[8- (β
-D-xylopyranosyloxy) nonanoyl] -N
([Epsilon])-[3- (4-hydroxyphenyl) propanoyl] -L-lysine hydrobromide (68h) 8- (methoxycarbonyl) octyl- [beta] -D- Xylopyranoside (3
8-) obtained by treating 0) with hydrazine monohydrate.
(Hydrazinocarbonyl) octyl-β-D-xylopyranoside is converted into acid azide, and this is converted to 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) by poly-L-lysine bromide, respectively. Hydrogenate (average molecular weight 11,700) (59). Post-treatment was carried out according to the procedure described in Reference Example 39 to obtain 128.7 mg of the complex (68h) as a colorless solid. Sugar content was measured by the phenol-sulfuric acid method. As a result, 8- (β-D-
It turned out that 10 mol of xylopyranosyloxy) nonanoyl groups had been introduced. Table 1 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 46 Poly-N (ε)-｛8- [4
-O- (β-D-galactopyranosyl) -β-D-glucopyranosyloxy] nonanoyl} -N (ε)-[3-
(4-Hydroxyphenyl) propanoyl] -L-lysine hydrobromide (68i) 8- (methoxycarbonyl) octyl-4-O- (β-
D-galactopyranosyl) -β-D-glucopyranoside (50) [J. Banoub and DR Bundle, Can. J. Chem.,
57 , 2085 (1979)] was treated with hydrazine monohydrate to give 8- (hydrazinocarbonyl) octyl-4-O- (β-D-galactopyranosyl) -β-. 128 mg (0.25 mmol) of D-glucopyranoside was dissolved in 1 ml of dry N, N-dimethylformamide, and 4N hydrochloric acid-dioxane solution 0.15
ml was added and cooled to -20 ° C. To this solution was added t-nitrite
28 mg of butyl dry N, N-dimethylformamide
2 ml was added and stirred for 30 minutes to obtain acid azide.
117 mg (1) of N-dimethylformamide solution was added to poly-L-lysine hydrobromide (molecular weight 11,700) (59).
0 micromolar) with 0.08 M sodium tetraborate (N
a ₂ B ₄ O ₇ ), 0.35 M potassium bicarbonate (KHC
O ₃₎ was added dropwise to the melted solution to 10 ml, and treated similarly to the procedure described in Reference Example 39, colorless solid as 128.
7 mg of the complex (68i) were obtained. Sugar content and 3- (4-
Table 1 shows the number of introduced hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 47 Poly-N (ε)-{4- [2
-(2- [2-acetamide-2-deoxy-4-O-
(Β-D-galactopyranosyl) -β-D-glucopyranosyloxy] ethoxy) ethoxy] butanoyl} -N
(Ε)-[3- (4-hydroxyphenyl) propanoyl] -L-lysine hydrobromide (68j) 2- [2- (3-methoxycarbonylpropyloxy)
2- [Ethoxy] ethyl-4-O- (β-D-galactopyranosyl) -2-acetamido-2-deoxy-β-D-glucopyranoside (43) obtained by treating with hydrazine monohydrate. 2- (3-hydrazinocarbonylpropyloxy) ethoxy] ethyl-4-O- (β-D-
(Galactopyranosyl) -2-acetamide-2-deoxy-β-D-glucopyranoside was converted to acid azide according to the procedure described in Reference Example 39, and this was converted to N-hydroxysuccinic acid 3- (4-hydroxyphenyl) propionate. The acid imide esters (62) were each coupled to poly-L-lysine hydrobromide (average molecular weight 11,700) (59). Post-treatment was performed according to Reference Example 39 to obtain 95 mg of a complex (68j) as a colorless solid. Sugar content was measured by the phenol-sulfuric acid method. As a result, it was found that 7 mol of sugar chains were introduced per 1 mol of poly-L-lysine. Table 1 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 48 Poly-N (ε)-[8- (2
-Acetamide-2-deoxy-6-O- (β-L-fucopyranosyl) -β-D-glucopyranosyloxy) nonanoyl] -N (ε)-[3- (4-hydroxyphenyl) propanoyl] -L -Lysine hydrobromide (68
k) 8- (methoxycarbonyl) octyl-2-acetamido-2-deoxy-6-O- (β-L-fucopyranosyl)
-Β-D-glucopyranoside (49) was converted to acid azide according to the procedure described in Reference Example 39, and this was converted to 3- (4
-Hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) was converted to poly-L-lysine hydrobromide (average molecular weight 11,700) (59)
And combined. Post-treatment was carried out according to Reference Example 39 to obtain 46 mg of the complex (68k) as a colorless solid. Sugar content was measured by the phenol-sulfuric acid method. As a result, poly
It was found that 12 moles of sugar chains were introduced per mole of L-lysine. Table 1 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 49 Poly-N (ε)-[8- (β
-D-mannopyranosyloxy) nonanoyl] -N
(Ε)-[3- (4-Hydroxyphenyl) propanoyl] -L-lysine hydrobromide (69a) According to the method described in Example 26, 8- (hydrazinocarbonyl) octyl-β-D- 145 mg (0.414 mmol) of mannopyranoside were dissolved in 3 ml of dry N, N-dimethylformamide, and the solution was dissolved in 4N hydrochloric acid under a stream of nitrogen.
0.6 ml of a dioxane solution was added, and a solution of t-butyl nitrite (90 mg) in dry N, N-dimethylformamide (0.6 ml) was added under cooling to -20 ° C, followed by stirring for 30 minutes to obtain acid azide. This solution was mixed with 150 mg (6.91 μmol) of poly-L-lysine hydrobromide (molecular weight 21,700) (60) in 0.08 M sodium tetraborate (Na ₂ B ₄ O).
₇ ) 0.35M potassium hydrogen carbonate (KHCO ₃ ) 10ml
At 4 to 5 ° C. Further 3-
9 mg (34.2 micromoles) of (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) in dry N, N-dimethylformamide 0.5
After dropping the ml solution, the mixture was stirred and reacted in a chamber at 4 ° C for 18 hours. By adding hydrobromic acid to the solution
The pH was set to 0.7 and 4 ° C. against pure water using a dialysis membrane of Spectra / Pore 3 (molecular weight cutoff = about 3,500).
Dialysis for 30 hours, take out the resulting dialysis solution, freeze-dry it, and obtain 220.7 mg of the complex (69
a) was obtained. Sugar content was measured by the phenol-sulfuric acid method. As a result, it was found that 43 mol of sugar chains were introduced per 1 mol of poly-L-lysine. Sugar content and 3-
Table 3 shows the number of introduced (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

[0093]

[Table 3]

Reference Example 50 Poly-N (ε)-[8- (β
-D-galactopyranosyloxy) nonanoyl] -N
(Ε)-[3- (4-Hydroxyphenyl) propanoyl] -L-lysine hydrobromide (69b) 8- (hydrazinocarbonyl) octyl-β-D-galactopyrase as in Reference Example 49 Is converted to an acid azide, which is combined with 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (6
2) was combined with poly-L-lysine hydrobromide (average molecular weight 21,700) (60) to form the complex (6)
9b) was obtained. Table 3 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 51 Poly-N (ε)-[8- (2
-Acetamide-2-deoxy-β-D-galactopyranosyloxy) nonanoyl] -N (ε)-[3- (4-hydroxyphenyl) propanoyl] -L-lysine hydrobromide (69c) Reference Example As in the case of 41, 8- (hydrazinocarbonyl) octyl-2-deoxy-2-acetamide-β-D
-Converting galactopyranoside to acid azide,
(4-Hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) was converted to poly-L
-Lysine hydrobromide (average molecular weight 21,700) (6
0) to give the complex (69c). Table 3 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 52 Poly-N (ε)-{4- [2
-(2- (2-Acetamide-2-deoxy-β-D-galactopyranosyloxy) ethoxy) ethoxy] butanoyl] -N (ε)-[3- (4-hydroxyphenyl)
Propanoyl] -L-lysine hydrobromide (69d) As in the case of Reference Example 42, 2- [2- (3-hydrazinocarbonylpropyloxy) ethoxy] ethyl-2-ethyl
Acetamide-2-deoxy-β-D-galactopyranoside is converted to acid azide, and 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) is converted to poly-L-lysine, respectively. Combined with hydrobromide (average molecular weight 21,700) (60),
A complex (69d) was obtained. Table 3 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 53 Poly-N (ε)-[8- (β
-L-fucopyranosyloxy) nonanoyl] -N (ε)
-[3- (4-hydroxyphenyl) propanoyl]-
L-Lysine hydrobromide (69e) In the same manner as in Reference Example 43, 8- (hydrazinocarbonyl) octyl-β-L-fucopyranoside was converted to acid azide, and this was converted to poly-L-lysine bromide. Hydrogenate (average molecular weight 21,700) was combined with (60), and further 3- (4
-Hydroxyphenyl) propionic acid was reacted with N-hydroxysuccinimide ester (62) to obtain a complex (69e). Table 3 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 54 Poly-N (ε)-[6- (2
-Acetamide-2-deoxy-α-D-mannopyranosyloxy) heptanoyl] -N (ε)-[3- (4-hydroxyphenyl) propanoyl] -L-lysine hydrobromide (69f) 8 -(Methoxycarbonyl) hexyl-2-acetamido-2-deoxy-α-D-mannopyranoside (35)
Was reacted with hydrazine monohydrate to give 6-
(Hydrazinocarbonyl) hexyl-2-acetamide-
Reference Example 39 was obtained using 2-deoxy-α-D-mannopyranoside.
And poly-L-lysine hydrobromide (average molecular weight 21,700) (60), and further 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) And a complex (69f) was obtained. In order to determine the sugar content, a portion was heated and stirred with 4N hydrochloric acid at 90-100 ° C for 6 hours.
When the obtained hydrolysis product was colorimetrically determined by the Elson-Mogen method, 6- (2-acetamido-2-deoxy-α) was obtained.
It was found that 28 mol of -D-mannopyranosyloxy) heptanoyl groups were introduced per mol of poly-L-lysine. Table 3 shows the number of 3- (4-hydroxyphenyl) propanoyl groups introduced. The physical constants of the compounds are shown in Table 2.

Reference Example 55 Poly-N (ε)-[8- (2
-Acetamide-2-deoxy-α-D-mannopyranosyloxy) nonanoyl] -N (ε)-[3- (4-hydroxyphenyl) propanoyl] -L-lysine hydrobromide (69 g) 8 -(Methoxycarbonyl) octyl-2-acetamido-2-deoxy-α-D-mannopyranoside (39)
Was reacted with hydrazine monohydrate to obtain 8-
(Hydrazinocarbonyl) octyl-2-acetamide-
Reference Example 39 was obtained using 2-deoxy-α-D-mannopyranoside.
And the poly (L-lysine hydrobromide) (average molecular weight 21,700) (60), and further 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62). ) To obtain a complex (69 g). Table 3 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 56 Poly-N (ε)-[8- (β
-D-xylopyranosyloxy) nonanoyl] -N
(Ε)-[3- (4-Hydroxyphenyl) propanoyl] -L-lysine hydrobromide (69h) As in the procedure described in Reference Example 39, 8- (methoxycarnibol) octyl-β-D Xylopyranoside (30)
Was treated with hydrazine monohydrate to convert 8- (hydrazinocarbonyl) octyl-β-D-xylopyranoside to acid azide, and poly-L-lysine hydrobromide (average molecular weight 21,700) (60), and further 3- (4-hydroxyphenyl) propionic acid N-
Reacted with hydroxysuccinimide ester (62). According to the operation described in Reference Example 39, post-processing is performed.
The complex (69h) was obtained as a colorless solid. Sugar content was measured by the phenol-sulfuric acid method. As a result, poly-L-
It was found that 29 moles of 8- (β-D-xylopyranosyloxy) nonanoyl groups were introduced per mole of lysine. Table 3 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 57 Poly-N (ε)-｛8- [4
-(1-β-D-galactopyranosyl) -β-D-glucopyranosyloxy] nonanoyl} -N (ε)-[3-
(4-Hydroxyphenyl) propanoyl] -L-lysine hydrobromide (69i) In the same manner as in the reaction procedure described in Reference Example 46, 8- (hydrazinocarbonyl) octyl-4- (1-β-D Acid azide obtained from (galactopyranosyl) -β-D-glucopyranoside was reacted with poly-L-lysine hydrobromide (average molecular weight 21,700) (60), and further reacted with 3- (4-
(Hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62). Reference Example 39
The compound (69i) was obtained as a colorless solid by post-treatment according to the method described in (1). Sugar content was measured by the phenol-sulfuric acid method. As a result, it was found that 53 mol of sugar chains were introduced per mol of poly-L-lysine. Table 3 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 58 Poly-N (ε)-{4- [2
-(2- [2-acetamide-2-deoxy-4-O-
(Β-D-galactopyranosyl) -β-D-glucopyranosyloxy] ethoxy) ethoxy] butanoyl} -N
(Ε)-[3- (4-Hydroxyphenyl) propanoyl] -L-lysine hydrobromide (69j) 2- [2- (3-) obtained by treating compound (43) with hydrazine monohydrate. Hydrazinocarbonylpropyloxy) ethoxy] ethyl-4-O- (β-D-galactopyranosyl) -2-acetamido-2-deoxy-β-D
-Glucopyranoside was converted to acid azide according to the procedure described in Reference Example 39, and bound to poly-L-lysine hydrobromide (average molecular weight 21,700) (60), followed by 3- (4-hydroxyphenyl) ) Propionic acid N-
Reacted with hydroxysuccinimide ester (62). According to Reference Example 39, post-treatment was carried out to give 7 as a colorless solid.
5 mg of the conjugate (69j) were obtained. Sugar content is phenol-
It was measured by the sulfuric acid method. As a result, poly-L-lysine 1
It was found that 13 moles of sugar chains were introduced per mole. Table 3 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 59 Poly-N (ε)-｛8- [6
-O- (α-D-mannopyranosyl) -α-D-mannopyranosyloxy] nonanoyl} -N (ε)-[3-
(4-Hydroxyphenyl) propanoyl] -L-lysine hydrobromide (69 l) 8- (methoxycarbonyl) octyl- [6-O- (α
-D-mannopyranosyl) -α-D-mannopyranoside (57) was converted to acid azide according to the procedure described in Reference Example 39, and poly-L-lysine hydrobromide (average molecular weight 21,700) (60 ) And then 3-
It was reacted with (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62). Post-treatment was carried out according to Reference Example 39 to obtain 95 mg of the complex (69 l) as a colorless solid. Sugar content was measured by the phenol-sulfuric acid method. As a result, it was found that 13 mol of sugar chains were introduced per 1 mol of poly-L-lysine. Table 3 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 60 Poly-N (ε)-｛8-
[3,6-di-O- (α-D-mannopyranosyl) -α
-D-mannopyranosyloxy] nonanoyl} -N
(Ε)-[3- (4-hydroxyphenyl) propanoyl] -L-lysine hydrobromide (69m) 8- (methoxycarbonyl) octyl-3,6-di-O
8- (hydrazinocarbonyloctyl-3,6-di-O- (α-D-mannopyranosyl) obtained by treating-(α-D-mannopyranosyl) -α-D-mannopyranoside (58) with hydrazine monohydrate. ) -Α-D-mannopyranoside is converted to acid azide, and this is converted to 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) by poly-L-lysine hydrobromide (average molecular weight) 21,700) (60).
Post-processing was performed according to Reference Example 39, and the complex (69
m). Sugar content was measured by the phenol-sulfuric acid method. As a result, it was found that 31 mol of sugar chains were introduced per 1 mol of poly-L-lysine. Sugar content and 3-
Table 3 shows the number of introduced (4-hydroxyphenyl) propanoyl groups. The physical constants of the compounds are shown in Table 2.

Reference Example 61 Poly-N (ε)-[8- (β
-D-mannopyranosyloxy) nonanoyl] -N
(Ε)-[3- (4-Hydroxyphenyl) propanoyl] -L-lysine hydrobromide (70a) According to the method described in Example 39, 8- (hydrazinocarbonyl) octyl-β-D- 180 mg (0.514 mmol) of mannopyranoside are converted to the acid azide in dry N, N-dimethylformamide and the solution is treated with poly-L-lysine hydrobromide (molecular weight 59,000).
(61) 300 mg (5.08 micromol) in 0.08
M sodium tetraborate _{(Na 2 B 4 O 7)} , 0.35M
It was added to a solution dissolved in 20 ml of potassium hydrogen carbonate (KHCO ₃ ). Next, 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62)
A solution of 12 mg (45.6 μmol) of dry N, N-dimethylformamide (0.5 ml) was added dropwise, and the mixture was stirred and reacted in a 4 ° C. chamber for 18 hours. The pH of the reaction solution was adjusted to 1.0 to 2.5 with concentrated hydrobromic acid, and 30% of pure water at 4 ° C using a dialysis membrane of Spectra / Pore 3 (molecular weight cut off: about 3,500). Dialyzed for hours. The obtained dialysis solution was taken out and freeze-dried to obtain 39% as a colorless solid.
1.9 mg of the complex (70a) were obtained. Colorimetric determination of the sugar content by the phenol-sulfuric acid method revealed that 100 moles of 8- (β-D-mannopyranosyloxy) nonanoyl groups were introduced per mole of poly-L-lysine. When the 3- (4-hydroxyphenyl) propanoyl group content was quantified by absorbance (276 nm), it was found that 9 moles had been introduced. Table 4 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. Table 5 shows physical constants of the compounds.

[0106]

[Table 4]

[0107]

[Table 5]

Reference Example 62 Poly-N (ε)-[8- (β
-D-galactopyranosyloxy) nonanoyl] -N
(Ε)-[3- (4-Hydroxyphenyl) propanoyl] -L-lysine hydrobromide (70b) 8- (hydrazinocarbonyl) octyl-β-D-galactopyrase as in Reference Example 49 The acid azide obtained from the noside and N-hydroxysuccinimide imide ester of 3- (4-hydroxyphenyl) propionic acid (62)
Poly-L-lysine hydrobromide (average molecular weight 59,00
0) (61) to obtain a complex (70b).
Table 4 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. Table 5 shows physical constants of the compounds.

Reference Example 63 Poly-N (ε)-[8- (2
-Acetamide-2-deoxy-β-D-galactopyranosyloxy) nonanoyl] -N (ε)-[3- (4-hydroxyphenyl) propanoyl] -L-lysine hydrobromide (70c) Reference Example As in the case of 41, 8- (hydrazinocarbonyl) octyl-2-deoxy-2-acetamide-β-D
-Converting galactopyranoside to acid azide,
(4-Hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) was converted to poly-L
-Lysine hydrobromide (average molecular weight 59,000) (6
0) to give the complex (70c). Table 4 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. Table 5 shows physical constants of the compounds.

Reference Example 64 Poly-N (ε)-{4- [2
-(2- (2-Acetamide-2-deoxy-β-D-galactopyranosyloxy) ethoxy) ethoxy] butanoyl} -N (ε)-[3- (4-hydroxyphenyl)
Propanoyl] -L-lysine hydrobromide (70d) As in Reference Example 42, 2- [2- (3-hydrazinocarbonylpropyloxy) ethoxy] ethyl-2-ethyl
Acid azide obtained from acetamide-2-deoxy-β-D-galactopyranoside and 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) were converted to poly-L-lysine bromide. The complex (7) was reacted with a hydrochloride (average molecular weight: 59,000) (61).
0d) was obtained. Table 4 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. Table 5 shows physical constants of the compounds.

Reference Example 65 Poly-N (ε)-[8- (β
-L-fucopyranosyloxy) nonanoyl] -N (ε)
-[3- (4-hydroxyphenyl) propanoyl]-
L-Lysine hydrobromide (70e) In the same manner as in Reference Example 30, 8- (hydrazinocarbonyl) octyl-β-L-fucopyranoside was converted to acid azide, and this was converted to poly-L-lysine bromide. Hydrogenate (average molecular weight 59,000) (61) and further combined with 3- (4
-Hydroxyphenyl) propionic acid was reacted with N-hydroxysuccinimide ester (62) to obtain a complex (70e). Table 4 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. Table 5 shows physical constants of the compounds.

Reference Example 66 Poly-N (ε)-[6- (2
-Acetamide-2-deoxy-α-D-mannopyranosyloxy) heptanoyl] -N (ε)-[3- (4-hydroxyphenyl) propanoyl] -L-lysine hydrobromide (70f) 6 -(Methoxycarbonyl) hexyl-2-acetamido-2-deoxy-α-D-mannopyranoside (35)
With hydrazine monohydrate to give 6-hydrazinocarbonylhexyl-2-acetamido-2-deoxy-α-D-mannopyranoside according to the method described in Reference Example 39, using poly-L-lysine hydrogen bromide. Acid salt (average molecular weight 59,000) (61), and further 3-
This was reacted with (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) to obtain a complex (70f). Partly 4N to determine sugar content
The mixture was heated and stirred with hydrochloric acid at 90-100 ° C for 6 hours. When the obtained hydrolysis product was colorimetrically determined by the Elson-Morgan method, 6- (2-acetamido-2-deoxy-α-D-mannopyranosyloxy) heptanoyl group was converted to 1 mol of poly-L-lysine. It was found that 45 moles were introduced per unit. Table 4 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. Table 5 shows physical constants of the compounds.

Reference Example 67 Poly-N (ε)-[8- (2
-Acetamide-2-deoxy-α-D-mannopyranosyloxy) nonanoyl] -N (ε)-[3- (4-hydroxyphenyl) propanoyl] -L-lysine hydrobromide (70 g) As in Example 55, 8- (methoxycarbonyl) octyl-2-acetamido-2-deoxy-α-
8- (Hydrazinocarbonyl) obtained by reacting D-mannopyranoside (39) with hydrazine monohydrate
Octyl-2-acetamide-2-deoxy-α-D-mannopyranoside was prepared according to the method described in Reference Example 39 using poly-L-lysine hydrobromide (average molecular weight 21,700).
(60) and further reacted with 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) to obtain a complex (70 g).
Table 4 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. Table 5 shows physical constants of the compounds.

Reference Example 68 Poly-N (ε)-[8- (β
-D-xylopyranosyloxy) nonanoyl] -N
(Ε)-[3- (4-Hydroxyphenyl) propanoyl] -L-lysine hydrobromide (70h) As in the procedure described in Reference example 39, 8- (methoxycarbonyl) octyl-β-D- Xylopyranoside (3
0) was treated with hydrazine monohydrate to give 8-
(Hydrazinocarbonyl) octyl-β-D-xylopyranoside is converted to acid azide, bound to poly-L-lysine hydrobromide (average molecular weight 59,000) (61), and further treated with 3- (4-hydroxy (Phenyl) propionic acid N-hydroxysuccinimide ester (62). Post-treatment was performed according to the operation described in Reference Example 39 to obtain a complex (70h) as a colorless solid. Sugar content was measured by the phenol-sulfuric acid method. As a result, it was found that 77 moles of 8- (β-D-xylopyranosyloxy) nonanoyl group was introduced per mole of poly-L-lysine. Table 4 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. Table 5 shows physical constants of the compounds.

Reference Example 69 Poly-N (ε)-｛8- [4
-(1-β-D-galactopyranosyl) -β-D-glucopyranosyloxy] nonanoyl} -N (ε)-[3-
(4-Hydroxyphenyl) propanoyl] -L-lysine hydrobromide (70i) In the same manner as in the reaction procedure described in Reference Example 46, 8- (hydrazinocarbonyl) octyl-4- (1-β-D -Galactopyranosyl) -β-D-glucopyranoside was reacted with poly-L-lysine hydrobromide (average molecular weight 59,000) (61), and further reacted with 3-
It was reacted with (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62). Post-treatment was carried out by the method described in Reference Example 39 to obtain a complex (70i) as a colorless solid. Sugar content is phenol-
It was measured by the sulfuric acid method. As a result, poly-L-lysine 1
It was found that 25 mol of sugar chains were introduced per mol. Table 4 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. Table 5 shows physical constants of the compounds.

Reference Example 70 Poly-N (ε)-｛8- [6
-O- (α-D-mannopyranosyl) -α-D-mannopyranosyloxy] nonanoyl} -N (ε)-[3-
(4-Hydroxyphenyl) propanoyl] -L-lysine hydrobromide (701) 8- (methoxycarbonyl) octyl-6-O- (α-
D-mannopyranosyl) -α-D-mannopyranoside (57) was converted to acid azide according to the procedure described in Reference Example 39, and poly-L-lysine hydrobromide (average molecular weight 59,000) (61) was obtained. And then combined with 3-
It was reacted with (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62). Post-treatment was carried out according to Reference Example 39 to obtain 95 mg of the complex (701) as a colorless solid. Sugar content was measured by the phenol-sulfuric acid method. As a result, it was found that 12 mol of sugar chains were introduced per 1 mol of poly-L-lysine. Table 4 shows the sugar content and the number of introduced 3- (4-hydroxyphenyl) propanoyl groups. Table 5 shows physical constants of the compounds.

Example 1 Poly-N (ε)-[8- (β-
D-mannopyranosyloxy) nonanoyl] -N (ε)
-[3- (4-hydroxyphenyl) propanoyl]-
N (ε)-[(2-methoxyethoxy) acetyl] -L
-Lysine (71a) 44.8 mg (0.128 mmol) of 8- (hydrazinocarbonyl) octyl-β-D-mannopyranoside is dissolved in 1 ml of dry N, N-dimethylformamide and 4N hydrochloric acid-dioxane solution under a nitrogen stream. Add 0.15 ml, -2
Cooled to 0 ° C. To this solution was added a solution of 28 mg of t-butyl nitrite in 0.2 ml of dry N, N-dimethylformamide and stirred for 30 minutes. After confirming that the raw materials had been consumed by thin-layer chromatography, 14 mg of sulfamic acid was added. A 0.2 ml solution of dry N, N-dimethylformamide was added and reacted. Poly-L-lysine hydrobromide (molecular weight 1
1,700) (59) 75 mg (6.41 micromol)
With 0.08 M sodium tetraborate (Na ₂ B ₄ O ₇ )
The solution was dissolved in 10 ml of 0.35 M potassium hydrogen carbonate (KHCO ₃ ), and the N, N-dimethylformamide solution of the acid azide was added dropwise at 4 to 5 ° C. Next, 8.4 mg (32 micromil) of 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester in dry N, N
-0.2 ml of dimethylformamide solution was added dropwise and then 4
The reaction was carried out by stirring in a chamber at a temperature of 18 ° C. for 18 hours. Subsequently, 161.4 mg of N-hydroxysuccinimide was added to a solution of 180 mg of 2-methoxyethoxyacetic acid (10) in 1 ml of dry N, N-dimethylformamide, and the mixture was stirred. 295 mg of (dimethylamino) propyl-carbodiimide hydrochloride (EDCHCl)
A solution of 2 ml of dry N, N-dimethylformamide was added to give 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (63). This solution was added to an aqueous solution of poly-L-lysine and reacted by stirring. This reaction was repeated twice to complete the reaction, and dialyzed against pure water at 4 ° C. for 30 hours using a dialysis membrane of Spectra / Pore 3 (molecular weight cut-off = about 3,500). Was taken out and freeze-dried to obtain 120 mg of the complex as a colorless solid. Further, this was applied to a column of Sephadex G100 (20
The solution was purified by gel filtration column chromatography eluting with 0.1M saline using a solution having a diameter of 50 mmφ. Each fraction eluted was subjected to high performance liquid chromatography <column: Tosoh, TSK Gel G3,000PW _XL , eluent:
0.1 M saline, elution rate: 0.4 ml / min, detector: U
V (280 nm)>, the target fractions were collected and dialyzed again against pure water at 4 ° C. for 30 hours. Subsequently, the mixture was freeze-dried to obtain the complex (71) as a colorless solid.
a) 95.7 mg were obtained. The number of residual amino groups in the complex was determined by the fluorescamine method [P. Bohlen, S. Stein, W. Dairman and
S. Udenfriend, Arch. Biochem. Biophys., 155,213-220
(1973)], determined by measuring the fluorescence intensity at 475 nm, with excitation at 390 nm. Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

[0118]

[Table 6]

[0119]

[Table 7]

Example 2 Poly-N (ε)-[8- (β-
D-galactopyranosyloxy) nonanoyl] -N
(Ε)-[3- (4-hydroxyphenyl) propanoyl] -N (ε)-[(2-methoxyethoxy) acetyl] -L-lysine (71b) 8- (hydrazinocarbonyl) octyl-β-D- 45 mg (0.128 mmol) of galactopyranoside was dissolved in 1 ml of dry N, N-dimethylformamide, and 0.15 ml of a 4N hydrochloric acid-dioxane solution was added under a stream of nitrogen to give -2.
Cooled to 0 ° C. To this solution was added a solution of 28 mg of t-butyl nitrite in 0.2 ml of dry N, N-dimethylformamide, and the mixture was stirred for 30 minutes to obtain an acid azide. Thereafter, the reaction was carried out in the same manner as in Example 1. . Further, the remaining amino group was 2-methoxyethoxyacetic acid (10) 280 mg,
240 mg of N-hydroxysuccinimide and 1,3-
2 obtained from 412 mg of dicyclohexylcarbodiimide.
-Methoxyethoxyacetic acid N-hydroxysuccinimide ester (63) in tetrahydrofuran was added to give 1
The reaction was performed for 8 hours. After completion of the reaction, the mixture was dialyzed using a dialysis membrane of Spectra / pore (molecular weight cut off = about 3,500), and the obtained dialysate was freeze-dried to obtain a colorless solid of 13%.
3 mg of the complex (71b) were obtained. Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 3 Poly-N (ε)-[8- (2-
Acetamide-2-deoxy-β-D-galactopyranosyloxy) nonanoyl] -N (ε)-[3- (4-hydroxyphenyl) propanoyl] -N (ε)-[(2-
[Methoxyethoxy) acetyl] -L-lysine (71c) In a solution of 50 mg of the complex (68c) in 2 ml of water, a solution of 270 mg of 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (63) in 1 ml of tetrahydrofuran was added.
08M sodium tetraborate (Na ₂ B ₄ O ₇ ) and 0.1M sodium tetraborate.
8M solution of 35M potassium bicarbonate (KHCO ₃ ) was added alternately, and the mixture was stirred at room temperature for 18 hours. The same post-treatment as in Example 1 was performed to obtain 49.3 mg of a complex (71c). Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 4 Poly-N (ε)-[8- (β-
L-fucopyranosyloxy) noninol]-[3- (4
-Hydroxyphenyl) propanoyl] -N (ε)-
[(2-Methoxyethoxy) acetyl] -L-lysine (71e) According to Example 1, conjugate (68e) was converted to 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (6
By reacting with 3), a complex (71e) was obtained. Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 5 Poly-N (ε)-[8- (2-
Acetamide-2-deoxy-α-D-mannopyranosyloxy) nonanoyl] -N (ε)-[3- (4-hydroxyphenyl) propanoyl] -N (ε)-[(2-methoxyethoxy) acetyl ] -L-Lysine (71 g) The conjugate (68 g) was prepared according to Example 1 by converting 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (6
By reacting with 3), a complex (71 g) was obtained. Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 6 Poly-N (ε)-[8- (β-
D-xylopyranosyloxy) nonanoyl] -N (ε)
-[3- (4-hydroxyphenyl) propanoyl]-
N (ε)-[(2-methoxyethoxy) acetyl] -L
-Lysine (71h) The conjugate (68h) was treated with 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (63) according to Example 1.
And a complex (71h) was obtained. Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 7 Poly-N (ε)-{8- [4-
(1-β-D-galactopyranosyl) -β-D-glucopyranosyloxy] nonanoyl} -N (ε)-[3-
(4-hydroxyphenyl) propanoyl] -N (ε)
-[(2-methoxyethoxy) acetyl] -L-lysine (71i) According to Example 1, the conjugate (68i) was converted to 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (63)
And a complex (71i) was obtained. Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 8 Poly-N (ε)-{8- [3,
6-di-O- (α-D-mannopyranosyl) -α-D-
[Mannopyranosyloxy] nonanoyl} -N (ε)-
[3- (4-hydroxyphenyl) propanoyl] -N
(Ε)-[(2-methoxyethoxy) acetyl] -L-
Lysine (71m) The conjugate (68m) was prepared according to Example 1 using 2-methoxyethoxyacetic acid N-hydroxysuccinimide (63).
Was reacted to obtain a complex (71 m). Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 9 Poly-N (ε)-[8- (2-
Acetamide-2-deoxy-6-O- (β-L-fucopyranosyl) -β-D-glucopyranosyloxy) nonanoyl] -N (ε)-[3- (4-hydroxyphenyl)
Propanoyl] -N (ε)-[(2-methoxyethoxy) acetyl] -L-lysine (71k) According to Example 1, the complex (68k) was converted to 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (63)
Was reacted to obtain a complex (71k). Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 10 Poly-N (ε)-[8- (β
-D-mannopyranosyloxy) nonanoyl] -N
(Ε)-[3- (4-hydroxyphenyl) propanoyl] -N (ε)-[(2-methoxyethoxy) acetyl] -L-lysine (72a) 2-methoxyethoxyacetic acid (10) 200 mg (1.49)
Mmol) and N-hydroxysuccinimide 60 mg
(0.52 mmol) in 2 ml of dry tetrahydrofuran was added to 1,3-dicyclohexylcarbodiimide (DC
C) A solution of 300 mg of 1 ml of dry tetrahydrofuran was added, and the mixture was stirred and reacted at room temperature for 18 hours. Thereafter, the precipitated dicyclohexylurea was filtered off to obtain 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (6
A solution of 3) in tetrahydrofuran was alternately mixed with 8 ml of 0.08 M sodium tetraborate (Na ₂ B ₄ O ₇ ) and 8 ml of 0.35 M potassium hydrogen carbonate (KHCO ₃ ).
9a) A solution of 70 mg (2.19 μmol) in 2 ml of pure water was added and stirred in a chamber at 4 ° C. overnight. This is 2
After repeating the reaction twice, the insoluble matter of the reaction solution was precipitated by a centrifugal separator, further dialyzed, and lyophilized to obtain a crude complex 67.3.
mg was obtained. Purification was performed in the same manner as in Example 1 to obtain 51.4 mg of the complex (72a). Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 11 Poly-N (ε)-[8- (β
-D-galactopyranosyloxy) nonanoyl] -N
(Ε)-[3- (4-hydroxyphenyl) propanoyl] -N (ε)-[(2-methoxyethoxy) acetyl] -L-lysine (72b) In the same manner as in Example 10, the complex (69b ) Was reacted with 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (63) to obtain a complex (72b). Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 12 Poly-N (ε)-[8- (2
-Acetamido-2-deoxy-β-D-galactopyranosyloxy) nonanoyl] -N (ε)-[3- (4-hydroxyphenyl) propanoyl] -N (ε)-[(2
-Methoxyethoxy) acetyl] -L-lysine (72
c) In the same manner as in Example 10, the complex (69c) was reacted with 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (63) to obtain a complex (72c). Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 13 Poly-N (ε)-[8- (β
-L-fucopyranosyloxy) nonanoyl]-[3-
(4-hydroxyphenyl) propanoyl] -N (ε)
-[(2-Methoxyethoxy) acetyl] -L-lysine (72e) In the same manner as in Example 10, the complex (69e) was reacted with 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (63). Thus, a composite (72e) was obtained. Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 14 Poly-N (ε)-[8- (2
-Acetamido-2-deoxy-α-D-mannopyranosyloxy) nonanoyl] -N (ε)-[3- (4-hydroxyphenyl) propanoyl] -N (ε)-[(2-
[Methoxyethoxy) acetyl] -L-lysine (72 g) In the same manner as in Example 10, the complex (69 g) was reacted with 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (63) to give a complex. (72 g) was obtained. Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 15 Poly-N (ε)-[8- (β
-D-xylopyranosyloxy) nonanoyl] -N
(Ε)-[3- (4-hydroxyphenyl) propanoyl] -N (ε)-[(2-methoxyethoxy) acetyl] -L-lysine (72h) In the same manner as in Example 10, the complex (69h ) Was reacted with 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (63) to obtain a complex (72h). Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 16 Poly-N (ε)-｛8- [4
-(1-β-D-galactopyranosyl) -β-D-glucopyranosyloxy] nonanoyl} -N (ε)-[3-
(4-hydroxyphenyl) propanoyl] -N (ε)
-[(2-methoxyethoxy) acetyl] -L-lysine (72i) Reaction of conjugate (69i) with 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (63) in the same manner as in Example 10. Thus, a complex (72i) was obtained. Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 17 Poly-N (ε)-[8- (β
-D-mannopyranosyloxy) nonanoyl] -N
(Ε)-[3- (4-hydroxyphenyl) propanoyl] -N (ε)-[(2-methoxyethoxy) acetyl] -L-lysine (73a) 70 mg (0.769 μmol) of the complex (70a) Was reacted with 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (63). Post-treatment was carried out in the same manner as in Example 10 to obtain 56 mg of the complex (73a). Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 18 Poly-N (ε)-[8- (β
-D-galactopyranosyloxy) nonanoyl] -N
(Ε)-[3- (4-hydroxyphenyl) propanoyl] -N (ε)-[(2-methoxyethoxy) acetyl] -L-lysine (73b) Complex (70b) 70 mg (0.769 μmol) Was reacted with 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (63). Post-treatment was carried out in the same manner as in Example 10 to obtain 35 mg of the complex (73a). Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 19 Poly-N (ε)-[8- (2
-Acetamido-2-deoxy-β-D-galactopyranosyloxy) nonanoyl] -N (ε)-[3- (4-hydroxyphenyl) propanoyl] -N (ε)-[(2
-Methoxyethoxy) acetyl] -L-lysine (73
c) 50 mg of the complex (70c) was treated with 2-methoxyethoxyacetic acid N
-Reacted with hydroxysuccinimide ester (63). Post-processing was performed in the same manner as in Example 10 to obtain 5
7 mg of the complex (73c) was obtained. Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound. Table 7 shows the physical constants.

Example 20 Poly-N (ε)-[8- (β
-L-fucopyranosyloxy) nonanoyl] -N (ε)
-[3- (4-hydroxyphenyl) propanoyl]-
N (ε)-[(2-methoxyethoxy) acetyl] -L
-Lysine (73e) The complex (70e) was reacted with 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (63).
Post-treatment was carried out in the same manner as in Example 10 to obtain a composite (73c). Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound.
Table 7 shows the physical constants.

Example 21 Poly-N (ε)-[8- (2
-Acetamido-2-deoxy-α-D-mannopyranosyloxy) nonanoyl] -N (ε)-[3- (4-hydroxyphenyl) propanoyl] -N (ε)-[(2-
[Methoxyethoxy) acetyl] -L-lysine (73 g) The conjugate (70 g) was reacted with 2-methoxyethoxyacetic acid N-hydroxysuccinimide ester (63).
Post-treatment was carried out in the same manner as in Example 10 to obtain a complex (73 g). Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound.
Table 7 shows the physical constants.

Example 22 Poly-N (ε)-[8- (β
-D-xylopyranosyloxy) nonanoyl] -N
(Ε)-[3- (4-Hydroxyphenyl) propanoyl] -N (ε)-[(2-methoxyethoxy) acetyl] -L-lysine (73h) Complex (70h) was converted to 2-methoxyethoxyacetic acid N- Reaction with hydroxysuccinimide ester (63).
Post-treatment was carried out in the same manner as in Example 10 to obtain a complex (73h). Table 6 shows the number of residual amino groups and the number of (2-methoxyethoxy) acetyl groups introduced in the compound.
Table 7 shows the physical constants.

Reference Example 71 Poly-ε- [3- (4-hydroxyphenyl) propanoyl] -ε- [2-methoxyethoxyacetyl) -L-lysine (78) Poly-L-lysine hydrobromide (average) Molecular weight 21,40
0) 250 mg (20.5 micromoles)
Sodium tetraborate (Na ₂ B ₄ O ₇ ) and 0.35M
Dissolve in 15 ml of potassium hydrogen carbonate (KHCO ₃ ) solution,
3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) 1 at 4-5 ° C
A solution of 5 mg (57.0 micromol) of 0.2 ml of dry N, N-dimethylformamide was added dropwise, and the mixture was stirred in a 4 ° C champer for 5 hours. A solution of 750 mg (5.48 mmol) of 2-methoxyethoxyacetic acid (10) and 638 mg (5.54 mmol) of N-hydroxysuccinimide in a 2 mL solution of dry N, N-dimethylformamide was dissolved in ice-cooled 1,3- 530mg of dicyclohexylcarbodiimide
10 ml of dry tetrahydrofuran (2.78 mmol)
After adding the solution, the solution of 2-methoxyethoxyacetic acid N-hydroxysuccinimide active ester (63) prepared by stirring overnight at room temperature was filtered, and the filtrate was added to the reaction solution. Thereafter, the mixture was stirred in a chamber at 4 ° C. for 18 hours. The reaction solution was analyzed using Spectra / Pore 1 (molecular weight cutoff =
(About 6,000) using a dialysis membrane. The obtained in-dialysis solution was subjected to centrifugation to precipitate insolubles. The supernatant was applied to a column of Sephadex G100 (20 mmφ × 50
cm) and purified by gel filtration column chromatography eluting with 0.1 M saline. Fractions of the target substance were collected and dialyzed again against pure water at 4 ° C. for 30 hours. The colorless solid obtained by freeze-drying the dialysis solution was re-
Methoxyethoxyacetic acid N-hydroxysuccinimide active ester (63) was reacted and subjected to the same post-treatment to obtain 90 mg of a colorless solid complex (78). The number of remaining amino groups determined by the fluorescamine method was 6. 278 nm
The number of 3- (4-hydroxyphenyl) propionic acids determined by the absorbance was 6. ^{_{78: [α] D 29 -}} 70.0 ° (c 0.12, H 2 O) IR (KBr): ν max cm -1: 3400, 2933, 1657, 1545

Example 23 Poly-ε- [8- (β-D-
Galactopyranosyloxy) octylcarbonyl] -ε
-[3- (4-hydroxyphenyl) propanoyl]-
L-Lysine hydrobromide (79, 80) (a) 8- (methoxycarbonyl) octyl β-D-
Galactopyranoside (15) was treated with hydrazine monohydrate to obtain 72 mg (0.2 mmol) of 8- (hydrazinocarbonyl) octyl β-D-galactopyranoside in 2 ml of dry N, N-dimethylformamide. , And 0.2 ml of 4N hydrochloric acid-dioxane solution was added.
Cooled to ° C. To this solution is added tert-butyl nitrite 0.02
85 ml was added and stirred for 30 minutes to obtain acid azide. The N, N-dimethylformamide solution was added to 150 mg of poly-L-lysine hydrobromide (average molecular weight 21,400).
(6.9 micromoles) was added dropwise to a solution of 15 mL of 0.08 M sodium tetraborate (Na ₂ B ₄ O ₇ ) and 0.35 M potassium bicarbonate (KHCO ₃ ) solution. Next, a solution of 8 mg of 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62) in 0.1 ml of dry N, N-dimethylformamide was added dropwise, followed by stirring in a 4 ° C. chamber for 17 hours. . The reaction solution was treated with Spectra / Pore 1 (molecular weight cutoff = about 6,0
Dialysis was performed using the dialysis membrane of (00). The dialysate was freeze-dried to obtain a colorless solid complex (79). The number of galactose per molecule determined by the phenol-sulfuric acid method is 24
Met. ^{_{79: [α] D 20 -}} 18.7 ° (c 0.25, H 2 O) IR (KBr): ν max cm -1: 3300, 3069, 2930, 2867, 1650,
1547

(B) 8- (Methoxycarbonyl) octyl β-D-galactopyranoside (15) 8- (hydrazinocarbonyl) octyl β-D-galactopyrano obtained by treating hydrazine monohydrate Sid 36mg
(0.1 mmol) was dissolved in 2 ml of dry N, N-dimethylformamide, 0.1 ml of a 4N hydrochloric acid-dioxane solution was added, and the mixture was cooled to -20 ° C. To this solution was added 0.014 ml of tert-butyl nitrite, and the mixture was stirred for 30 minutes to form an acid azide. The N, N-dimethylformamide solution was added to poly-
L-Lysine hydrobromide (average molecular weight 21,400) 1
50 mg (12.8 μmol) was added dropwise to a solution of 15 ml of 0.08 M sodium tetraborate (Na ₂ B ₄ O ₇ ) and 0.35 M potassium hydrogen carbonate (KHCO ₃ ) solution. Next, 3- (4-hydroxyphenyl) propionic acid N-hydroxysuccinimide ester (62)
After 8 mg of a solution of 0.1 ml of dry N, N-dimethylformamide was added dropwise, the mixture was stirred in a chamber at 4 ° C. for 18 hours. The reaction solution was analyzed using Spectra / Pore 1 (molecular weight cutoff =
(About 6,000) using a dialysis membrane. The dialysis solution was freeze-dried to obtain a colorless solid complex (80).
The number of galactose per molecule determined by the phenol-sulfuric acid method was 12. ^{_{80: [α] D 20 -}} 36.1 ° (c 0.21, H 2 O) IR (KBr): ν max cm -1: 3296, 2937, 1650, 1545

Example 24 Poly-ε- [8- (β-D-
Galactopyranosyloxy) octylcarbonyl] -ε
-[3- (4-hydroxyphenyl) propanoyl]-
N-ε-[(2-methoxyethoxy) acetyl] -L-
Lysine (81, 82) (a) To 2 ml of the aqueous solution of the galactose-containing polylysine (79) obtained in Example 23 (a), 500 mg of 2-methoxyethoxyacetic acid (10) and N-hydroxysuccinimide 4
A solution of 2-methoxyethoxyacetic acid N-hydroxysuccinimide (63) obtained from 25 mg and 760 mg of 1,3-dicyclohexylcarbodiimide in tetrahydrofuran and 0.08 M sodium tetraborate (Na ₂ B)
₄ O ₇ ) and 0.35 M potassium bicarbonate (KHCO
₃ ) 15 ml of the solution was added and reacted for 18 hours. The reaction solution was treated with Spectra / Pore 1 (molecular weight cut off = about 6,000).
Dialysis was performed using the dialysis membrane of 0). A colorless solid obtained by freeze-drying the dialyzed solution was purified by gel filtration column chromatography using a Sephadex G100 column (20 mmφ × 50 cm), eluting with 0.1 M saline. The fractions of the target substance are collected and again in pure water at 4 ° C. for 30 minutes.
Time dialysis was performed. The colorless solid obtained by freeze-drying the inner dialysate was reacted again with 2-methoxyethoxyacetic acid N-hydroxysuccinimide active ester (63) to obtain 56 mg of a colorless solid complex (81). The number of residual amino groups determined by the fluorescamine method was 13. The number of 3- (4-hydroxyphenyl) propionic acids determined by the absorbance at 278 nm was 3. ^{_{81: [α] D 29 -}} 54.6 ° (c 0.13, H 2 O) IR (KBr): ν max cm -1: 3301, 2930, 2864, 1655, 1547

(B) The aqueous solution of the galactose-containing polylysine (80) obtained in Example 23 (b) was treated in the same manner as in the above (a) to obtain 71 mg of a colorless solid complex (82).
The number of remaining amino groups was 10. The number of 3- (4-hydroxyphenyl) prokionic acids was 3. ^{_{82: [α] D 29 -}} 14.8 ° (c 0.50, H 2 O) IR (KBr): ν max cm -1: 3300, 2930, 2866, 1650, 1547

Example 25 Introduction of Methotrexate into Mannose-Containing Polylysine (Average Degree of Polymerization 282) To a solution of 5 mg of methotrexate in 0.5 ml of DMF was added 2.1 mg of dicyclocarbodiimide, and the mixture was stirred at 4 ° C. for 17.5 hours. 1.15 mg of hydroxysuccinimide was added, and the mixture was stirred and reacted at room temperature for 5 hours. After the reaction solution was filtered, the filtrate was filtered using the poly-N (ε)-obtained in Reference Example 61.
[8- (β-D-mannopyranosyloxy) nonanoyl] -N (ε)-[3- (4-hydroxyphenyl) propanoyl] -L-lysine hydrobromide (70a) 30
mg of water in 1 ml of water was added to 0.08 M sodium tetraborate (N
a ₂ B ₄ O ₇ ), 0.35 M potassium bicarbonate (KHC
O ₃ ) were alternately added dropwise and stirred at room temperature overnight to react. The pH was adjusted to 1.5 by adding concentrated hydrobromic acid to the reaction solution, and the spectrum / pore 3 (molecular weight cut off =
Dialysis was performed overnight against pure water using a dialysis membrane of about 3,500). The obtained dialysis solution was taken out and freeze-dried to obtain a yellow amorphous substance (74). ^{_{74: [α] D 28 -}} 11.4 ° _{(c = 0.11, H 2 O} ) IR (KBr) ν max cm -1: 3320, 1629, 1544, 1450, 1246, 1
50 mg of 2-methoxyethoxyacetic acid and 43 mg of N-hydroxysuccinimide were dissolved in 1 ml of dry DMF and cooled with ice.
To this solution, 76 mg of DCC was added with stirring, and the mixture was stirred and reacted at room temperature overnight. After the reaction solution was filtered, the filtrate was added to a solution of mannose-methotrexate-containing polylysine (74) obtained above in 4 ml of water, and 0.08 M sodium tetraborate (Na ₂ B ₄ O ₇ ) and 0.35 M potassium bicarbonate were added. (KHCO ₃ ) was alternately added dropwise with 4 ml, and the mixture was stirred and reacted at room temperature for 22 hours. After dialysis of the reaction solution, it was freeze-dried. 0.1 M saline was added to the obtained colorless amorphous substance, and the mixture was centrifuged. The supernatant was purified with a Sephadex G-100 column. The fraction containing polylysine was dialyzed for 2 days and freeze-dried to give 18 mg of yellow amorphous substance (75).
Obtained. Fluorescamine method [P. Bohlen, S. Stein, W. Dai
rman and S. Udenfriend, Arch.Biochem.Biophys., 15
5,213-220 (1973)], the number of remaining amino groups was calculated to be 10. Absorbance (360 nm) of methotrexate content
And it was found that 1 mol was introduced. 75: [α] _D ²⁸ + 3.0 ゜ (c = 0.20, H ₂ O) IR (KBr) ν _max cm ^-1 : 3510, 1640, 1384, 1116

Example 26 Introduction of Methotrexate into Mannose-Containing Polylysine (Average Degree of Polymerization 104) To a solution of 10 mg of methotrexate in 0.5 ml of DMF was added 4.5 mg of dicyclocarbodiimide, and the mixture was stirred at 4 ° C. for 18 hours. 2.3 mg of succinimide was added, and the mixture was stirred and reacted at room temperature for 5 hours. After the reaction solution was filtered, the filtrate was subjected to poly-N (ε)-[8-
(Β-D-mannopyranosyloxy) nonanoyl] -N
(Ε)-[3- (4-hydroxyphenyl) propanoyl] -L-lysine hydrobromide (69a) 20 mg water 1
Add 0.08 M sodium tetraborate (Na ₂ B ₄
O ₇ ), 0.35 M potassium bicarbonate (KHCO ₃ ) 3
The mixture was added dropwise to the mixture alternately with ml and stirred at room temperature overnight to react. The pH was adjusted to 1.5 by adding concentrated hydrobromic acid to the reaction solution, and the spectrum / pore 3 (molecular weight cutoff = about 3,5
00) was dialyzed against pure water overnight using a dialysis membrane. The obtained dialysis solution was taken out and freeze-dried to obtain a yellow amorphous substance (76). ^{_{76: [α] D 28 -}} 15.7 ° (c = 0.14, DMF) IR (KBr) ν max cm -1: 3330, 1676, 1540, 1414, 1352, 1
100 mg of 142,1000 2-methoxyethoxyacetic acid and 86 mg of N-hydroxysuccinimide were dissolved in 1 ml of dry DMF and cooled with ice. To this solution was added, with stirring, 152 mg of DCC.
The reaction was stirred overnight at room temperature. After filtering the reaction solution,
The filtrate was alternately added to a solution of mannose-methotrexate-containing polylysine (76) obtained above in 3 ml of water, alternately with 4 ml of 0.08 M sodium tetraborate (Na ₂ B ₄ O ₇ ) and 0.35 M potassium hydrogen carbonate (KHCO ₃ ). At room temperature.
The mixture was reacted by stirring for 2 hours. The reaction solution was dialyzed using a dialysis membrane of Spectra / pore (molecular weight cut-off = about 3,500) and freeze-dried. 0.1 M saline was added to the obtained yellow amorphous substance, and the mixture was centrifuged. The supernatant was purified with a Sephadex G-100 column. The fraction containing polylysine was dialyzed for 2 days and lyophilized to obtain 20 mg of a yellow amorphous substance (77). Fluorescamine method [P. Bo
hlen, S.Stein, W.Dairman and S.Udenfriend, Arch. Bi
ochem. Biophys., 155, 213-220 (1973)], the number of remaining amino groups was calculated to be 12. The methotrexate content was quantified by using the absorbance (360 nm), and it was found that 2 mol was introduced. ^{_{77: [α] D 28 -}} 3.90 ° _{(c = 0.13, H 2 O} ) IR (KBr) ν max cm -1: 1647, 1558, 1384, 1161, 1042

Reference Example 72 Polylysine (average degree of polymerization: 10
4) Introduction of Methotrexate to (84, 85) (a) To a solution of 175 mg of methotrexate in 1.75 ml of DMF, 60 mg of 1,3-dicyclohexylcarbodiimide was added.
1.5 ml of a DMF solution was added, and the mixture was stirred at 4 ° C. for 19.5 hours. Then, 42 mg of N-hydroxysuccinimide and 0.053 ml of pyridine were added, and the mixture was stirred at room temperature for 3.5 hours. After filtering 0.25 ml of the reaction solution, the filtrate was poly-L-
Lysine hydrobromide (average molecular weight 21,400) 100
mg of 0.08 M sodium tetraborate (Na ₂ B ₄ O ₇ )
It was added dropwise to a solution of 0.35M and potassium hydrogen carbonate (KHCO ₃ ), and the mixture was stirred at room temperature for 19.5 hours. The reaction solution was dialyzed overnight against pure water using a dialysis membrane of Spectra / Pore 1 (molecular weight cut off = about 6,000). The obtained dialysis solution was taken out and lyophilized to give a yellow amorphous solid (8
3, r = 2). 250 mg of methoxyethoxyacetic acid and 215 mg of N-hydroxysuccinimide were dried in DMF5
The mixture was dissolved in ml and cooled on ice. DCC3 is added to this solution while stirring.
80 mg was added, and the mixture was stirred at room temperature overnight. After filtering half of the reaction solution, the filtrate was added to 4 ml of the aqueous solution of methotrexate-containing polylysine obtained above, and 0.08 M sodium tetraborate (Na ₂ B ₄ O ₇ ) and 0.35 M potassium hydrogen carbonate (KHCO ₃ ) were added. 10 ml were alternately added dropwise and stirred at room temperature for 24 hours. Further, the same amount of methoxyethoxyacetic acid N-hydroxysuccinimide active ester was added, and the mixture was added at room temperature for 2 hours.
Stir for 5 hours. After dialysis of the reaction solution, it was freeze-dried.
After 0.1 M saline was added to the obtained colorless amorphous solid and centrifuged, the supernatant was purified with a Sephadex G-100 column. The fraction containing polylysine was dialyzed and freeze-dried to obtain 30 mg of a yellow amorphous complex (84). By the fluorescamine method, the number of remaining amino groups is 16
Was calculated. The methotrexate content was measured at an absorbance of 377.
Quantification using nm revealed that 2 mol of poly-L-lysine was introduced per 1 mol. ^{_{84: [α] D 28 -}} 12.2 ° _{(c 0.15, H 2 O)} IR (KBr) ν max cm -1: 3450, 1650, 1545, 1110

(B) A yellow amorphous complex (85) was obtained from compound (83, r = 4) in the same manner as in (a) except that the reaction time was changed. The number of amino groups was calculated to be 18. It was found that the content of methotrexate was 4 mol per mol of poly-L-lysine. ^{_{85: [α] D 28 -}} 11.1 ° _{(c 0.13, H 2 O)} IR (KBr) ν max cm -1: 3320, 1630, 1545, 1110

Example 27 Introduction of galactose into methotrexate-containing polylysine (average degree of polymerization 104) (8
8, 89) (a) 8- (Methoxycarbonyl) octyl β-D-galactopyranoside (15) obtained by treating hydrazine monohydrate with 8- (hydrazinocarbonyl) octyl β-D-galactopyr. 50 mg of noside is dried N, N-
It was dissolved in 2 ml of dimethylformamide, 0.15 ml of a 4N hydrochloric acid-dioxane solution was added, and the mixture was cooled to -20 ° C. To this solution was added 0.025 ml of tert-butyl nitrite to give 30
Stir for a minute to acid azide, filter half of the reaction,
The filtrate was treated with 0.08 M sodium tetraborate (Na ₂ B ₄ ) of the methotrexate-containing polylysine (83) obtained above.
O ₇ ) and 0.35M potassium bicarbonate (KHCO ₃ )
The solution was added dropwise and stirred at room temperature for 20 hours. After dialyzing the reaction solution, it was freeze-dried to obtain (86). The number of galactose per molecule determined by the phenol-sulfuric acid method was 47. 250 mg of 2-methoxyethoxyacetic acid and 215 mg of N-hydroxysuccinimide were dissolved in 5 ml of dry DMF and cooled with ice. 380 mg of 1,3-dicyclohexylcarbodiimide was added to this solution with stirring, and the mixture was stirred at room temperature for 19 hours. After filtering half of the reaction solution, the filtrate was washed with the aqueous solution 4 of methotrexate-containing polylysine (86) obtained above.
ml of 0.08 M sodium tetraborate (Na ₂ B ₄ O
₇ ) and 10 ml of a 0.35 M potassium hydrogen carbonate (KHCO ₃ ) solution were alternately added dropwise, and the mixture was stirred at room temperature for 24 hours. Further, the same amount of 2-methoxyethoxyacetic acid N-hydroxysuccinimide active ester (63) was added, and the mixture was added at room temperature for 2 hours.
Stir for 5 hours. After dialysis of the reaction solution, it was freeze-dried.
0.1 M saline was added to the obtained yellow amorphous solid, and the mixture was centrifuged. The supernatant was purified with a Sephadex G-100 column. The fraction containing polylysine was dialyzed and freeze-dried to obtain 10 mg of a yellow amorphous (88). The number of remaining amino groups was calculated to be 20 by the fluorescamine method. When the methotrexate content was quantified by using an absorbance of 377 nm, it was found that 3 mol was introduced per 1 mol of poly-L-lysine. ^{_{88: [α] D 28 -}} 11.4 ° _{(c 0.11, H 2 O)} IR (KBr) ν max cm -1: 3320, 1629, 1544, 1245, 1090

(B) By carrying out in the same manner as in the above (a), a compound (89) was obtained from the compound (87) having 26 galactose molecules per molecule. The number of remaining amino groups is 18
It was found that 4 mol of the methotrexate content was introduced per 1 mol of poly-L-lysine. ^{_{89: [α] D 28 -}} 13.8 ° _{(c 0.11, H 2 O)} IR (KBr) ν max cm -1: 3320, 1630, 1450, 1080

Experimental Example 1 Samples Sample 1 was a polylysine derivative (67) modified with ethylene glycol obtained in Reference Example 38, and Sample 2 was a polylysine derivative (71) modified with ethylene glycol and N-acetylgalactosamine obtained in Example 3.
c) was prepared. Unmodified polylysine hydrobromide (64) was prepared as a control sample. These three samples
^A hydroxyphenylpropanoyl group has each been introduced for ¹²⁵ I labeling. 2. Methods The sample was labeled with ¹²⁵ I prior to the experiment. ^{The 125} I labeling was carried out by the chloramine T method as usual, and the reaction solution was subjected to gel filtration with a Sephadex G-25 column to obtain a polymer fraction. To this fraction was added 1% of bovine serum albumin and a non-labeled compound to prepare a 1 mg / ml administration solution, which was used in the following experiments. The pharmacokinetics were examined by administering a ¹²⁵ I-labeled sample or a control sample at 1 mg / kg into the femoral vein of a male SD rat (7 weeks old) at 0.5 mg / kg after administration.
Blood was collected from the jugular vein 1, 2, and 3 minutes later, and 5 minutes later, the whole blood was collected from the abdominal aorta and killed, and 11 major organs were removed. Blood was subjected to centrifugal separation of the plasma, and the removed organ was weighed as it was. The radioactivity was measured by a gamma counter, and the plasma concentration and the concentration in each organ were calculated. Similarly, the concentration in the main organs 1 hour after the administration of the sample or control sample was also measured. Each animal experiment was performed with three animals per group.

The degradability (stability) of macromolecules in plasma was determined by the measurement of ¹²⁵ I in 0.45 ml of plasma collected from rats.
0.05 ml of 1 mg / ml of the labeled sample or control sample was added,
Heated at 37 ° C. for 2 hours. After 0, 5, 10, 30, 60 and 120 minutes, take 0.05 ml each, add 0.4 ml of acetonitrile or 25% trichloroacetic acid, precipitate the polymer part, measure the precipitate and the radioactivity of the supernatant, The ratio of the remaining polymer was determined.

[0154] 3. Results and Discussion FIGS. 1, 2 and 3 show the concentration of each sample in the organ at 5 minutes and 60 minutes after administration. The control sample was widely distributed in various organs, disappeared slowly from each organ until 60 minutes later, and was considered to be non-specifically adhered to each organ. However, since Sample 1 accumulates almost exclusively in the kidney and disappears quickly, its adhesion to nonspecific organs such as the control sample has been improved. Furthermore, Sample 2 modified with sugar had less accumulation in the kidney than Sample 1, and its accumulation in the liver was likely to be greater. Since it disappeared quickly in the kidney but tended to accumulate only in the liver, It was found to be useful as a drug carrier to the liver.

FIG. 4 shows the change in plasma concentration up to 5 minutes after the administration. Despite the same dose, control samples show very low plasma concentrations from the beginning. This is compared with the non-specific attachment of the control sample to each organ, as described above with reference to FIG. On the other hand, sample 1
And the sample 2 both have a high initial concentration, and the phenomenon in the control sample is not observed. After the initial stage, the sample 2 rapidly disappears from the plasma, and the effect of sugar modification appears.

FIG. 5 shows the ratio of the polymer when heated with plasma in a test tube. The control sample is rapidly degraded and degraded to a low molecular weight.
Even after a minute, the polymer remained almost as a polymer, indicating that the stability in blood was improved as compared with the control sample.

Therefore, it was found that by modifying the ε-amino group of polylysine with a group containing ethylene glycol, stability in blood was improved and specific adhesion to organs could be avoided. Modification with a sugar can provide an organ-specific drug carrier.

[Brief description of the drawings]

FIG. 1 is a graph showing the concentration in a main organ of a control sample at 5 minutes and 60 minutes after intravenous administration.

FIG. 2 is a graph showing the concentration in main organs at 5 minutes and 60 minutes after intravenous administration of Sample 1.

FIG. 3 is a graph showing the concentration in main organs at 5 minutes and 60 minutes after intravenous administration of Sample 2.

FIG. 4 is a graph showing changes in plasma concentrations of various samples after intravenous administration.

FIG. 5 is a graph showing the change over time in the ratio of the high molecular fraction when various samples are heated with plasma.

 ────────────────────────────────────────────────── ─── Continuing from the front page (72) Inventor Akinori Gonmasa 3-8-7 Chiyoda, Kashiwa-shi, Chiba 208 Vinus Kashiwa 208

Claims (3)

(57) [Claims] 1. A sugar-modified polylysine derivative comprising the structural units (A), (B) and (C) represented by the following formula (I). Embedded image 中 In the formula, R ¹ represents a glycosyl group, and X represents the following formula (II). Embedded image R ² represents the following formula (III) or (IV). Embedded image R ³ represents a hydrogen atom or a drug residue. p, q and r represent the numbers of the structural units (A), (B) and (C), and the numbers are 1 to 200, 10 to 300 and 1 to 20, respectively. ｝ 2. R ¹ is D-mannopyranosyl, D-galactopyranosyl, L-fucopyranosyl, xylopyranosyl, 2-acetamido-2-deoxy-D-galactopyranosyl, 2-acetamido-2-deoxy-D- Mannopyranosyl, 2-acetamido-2-deoxy-D-glucopyranosyl, 4-galactopyranosyl-D-glucopyranosyl, 4-galactopyranosyl-2-acetamide-2
-Deoxy-D-glucopyranosyl, 2-acetamide-
2-deoxy-6-fucosyl-D-glucopyranosyl,
6-mannopyranosyl-D-mannopyranosyl, 3,6
The sugar-modified polylysine derivative according to claim 1, which is -di-mannopyranosyl-D-mannopyranosyl. ^3. The sugar-modified polylysine derivative according to claim 1, wherein R ³ is a drug residue derived from methotrexate.