JPS5928564B2 - Novel amino sugar polymer and its production method - Google Patents

Description

[Detailed description of the invention] The present invention is based on the formula: (However, n indicates an integer from 70 to 120.

) and a method for producing the same. Polymers of α-1, 6-linked glucose may be contaminated with microorganisms such as Leuconostoc.
LeucOnOstOemese
There is a dextran produced by P. interOides).

Dextran itself is useful as a plasma expander, but its sulfate ester (dextran sulfate) has a lipid serum clarifying effect and lowers blood cholesterol and triglycerides. It also has anti-hyaluronidase and fibrinolytic effects, and is known as an effective drug for treating hyperlipidemia and arteriosclerosis. On the other hand, heparin, a mucopolysaccharide present in animal tissues, has a strong anti-blood coagulation effect.
It has a wide range of physiological effects such as lipid serum clarifying action, and is characterized by having sulfated amino sugar units, such as the following units, in its molecule.

(However, in the formula, X represents H or SO3-.

) Therefore, if it is possible to synthesize polysaccharides and sulfated products thereof that have α-1,6-linked sugar chains such as dextran and also contain amino groups such as heparin in their molecules, various physiological functions can be expected. However, polysaccharides having free amino groups are not naturally produced, and there have been no reports to date on the artificial synthesis of polysaccharides by polymerization of amino sugar monosaccharides.

Therefore, in view of the above-mentioned purpose, the present inventors have conducted intensive research and have synthesized a polysaccharide having an α-1,6-linked sugar chain and containing an amino group such as heparin in its molecule. They were very successful and completed the present invention.

(However, in the formula, Bn represents a benzyl group, and n is the same as above.

), and is the first compound synthesized by the present inventors.

Below, the synthesis process is shown in diagrams, and specific examples thereof are shown.

(However, in the formula, Bz represents a benzoyl group, Ms represents a methanesulfonyl group, and Bn and .n are the same as above.

) Compound (2) can be easily obtained by treating 1,6-anthro-β-D-glucopyranose (1) derived from β-D-glucopyranose with benzoyl chloride [M. Thank you EtalCOl
lectiOnCzechOslOvlChem. C.O.
nlnlun. vOl. 26, 2542 (1961).

]. Compound (2) 217 is dissolved in 500 ml of dry pyridine and cooled on ice.

Methanesulfonyl chloride 19f7 was added dropwise while stirring, and the temperature was gradually returned to room temperature. After 3 hours, the mixture is poured into a large amount of ice water and stirred to precipitate crude crystals of compound (3).

After removing P, washing with water, drying, and recrystallizing from methanol, white crystals 17y (yield 68%) are obtained. Compound (3) 107
Dissolve in 50ml of chloroform and cool on ice.

A sodium methoxide solution prepared by dissolving metallic sodium 4y in 60 ml of methanol is added dropwise to this while stirring. After the mixture was left at room temperature overnight, it was neutralized with 5% hydrochloric acid and concentrated to dryness under reduced pressure. Dilute the residue with 20ml of acetone.
Extract twice and concentrate the extract under reduced pressure to obtain an oil. When purified by column chromatography on silica gel (developing solvent: chloroform-methanol, 100:1 v/v), a mixture of compounds (4-a) and (4-b) was obtained with a yield of 3.1y (yield: 97%).
%). Mixture 6 of compounds (4-a) and (4-b)
．． 77 was dissolved in 300 ml of dry tetrahydrofuran, and 2. sodium hydride (purity 60%) was added under ice cooling.
87 and stir for 30 minutes.

Next, benzyl bromide 167 is added and reacted at room temperature for 4 hours. Add ice-cooled saturated ammonium chloride aqueous solution,
After stirring for 0 min, extract 3 times with 200 ml of ether. The extract was dried over anhydrous magnesium sulfate, concentrated under reduced pressure, and the resulting oil was purified by silica gel column chromatography (developing solvent: benzene monoethyl acetate 20:1 v/v) to obtain an oily compound (5-a). A mixture of (5-b) and 107 (yield: 86%) is obtained. Compound (5-a) and (
The mixture 107 of 5-b) is dissolved in a mixed solvent of 200 ml of ethanol and 50 ml of saturated ammonium chloride water, 17.57 ml of sodium azide is added, and the mixture is heated under reflux for 60 hours while stirring.

After cooling, 500 ml of distilled water is added, ethanol is removed under reduced pressure, and the remaining aqueous solution is extracted with chloroform. The extract was dried over anhydrous magnesium sulfate and concentrated under reduced pressure to obtain an oil. When purified by column chromatography on silica gel (developing solvent: benzene-ether, 20:1 v/v), a mixture of compounds (6-a) and (6-b) was obtained with 9.67
(yield 81%). [Compounds (6-a) and (6-b
Physical properties of the mixture of compounds (6-a) and (6-b)
) 9.3y of the mixture was dissolved in 180ml of dry tetrahydrofuran.
1 and chill on ice.

Add 1.9 f of sodium hydride (purity 60%) and stir for 30 minutes, then add 11.57 g of benzyl bromide. After stirring at room temperature for 3 hours, ice-cooled saturated ammonium chloride aqueous solution 500 g
Add m1 and stir for an additional 30 minutes. This mixed solution is extracted with ether, and the extract is dried over anhydrous magnesium sulfate and concentrated under reduced pressure to obtain an oil. Silica gel chromatography (developing solvent, hexane-ethyl acetate 5:1v/
) to obtain oily compound (7) 87 (yield 68%)
get. It crystallizes if left standing and can be recrystallized from cyclohexane. [Physical Properties of Compound (7)] The thus obtained compound (7) was polymerized by the following method to obtain the azido sugar polymer (8) which is the starting material of the present invention.
).

1,6-Anthitro 3-azido-2,4-di0-benzyl-3-deoxy-β-D-glucopyranose (7)
(0.50y11.36mm01) to 10-5mmHg
The mixture was dried under high vacuum overnight and dissolved in methylene chloride (1.0 ml) previously dried over calcium hydride in a vacuum ampoule.

This solution was mixed at -60°C into phosphorus pentafluoride (0.068 mm01), benzoyl fluoride (15 μElO.l4 mmOl), and methylene chloride (0.3 ml), which had been reacted for 30 minutes in a polymerization tube. After reacting at -60°C for 23 hours, the polymerization ampoule was opened and methanol was poured into it. At this time, the polymer precipitates, so chloroform is added to it until the polymer is sufficiently dissolved, neutralized with an aqueous sodium bicarbonate solution, washed with water, and dried over anhydrous sodium sulfate. After removing the desiccant by f, liquid f is concentrated and petroleum benzine is added to reprecipitate it. The operations of dissolving, concentrating, and reprecipitating were further performed twice, and the solution was dissolved in benzene and freeze-dried to obtain a polymer (8-a) of 0.40937 (conversion rate 8).
1.9%). IR: 2130cm-1' HNMR: δ7.2-7.0 (IOH, .d) Aromatech), 5.2-4.2 (5H, .m, .H-
1,10CH2Ph), 4.0~3.2(5H,
．． m,. H-2, 4, 5, 6, 6'), 3.08 (IH
、． q, H- 3) 13CNMR: δ96.81 (IC
、． C-1), 78.57 (IC,.C-5), 75
．． 98(IC)C-2), 74.81(IC,-C
H2Ph), 72.32(IC)-CH2Ph), 70
．． 96(1C)C-4), 65.59(2C,.C
-3, C-6) Next, the reaction was carried out in the same manner as above while changing the amount of compound (7) used and the polymerization time. Table 1 shows the results.

In this way, the starting material (8) is obtained, which is reduced by the following method to obtain the amino sugar polymer (9) of the present invention.
get. The reduction of the polymer (8) is carried out by Verte reduction, ie with an alkali metal in liquid ammonia.

Alkali metals include potassium, sodium, and lithium. It is necessary to add six or more atoms of alkali metal per unit in the polymer, and it is usually desirable to add three times the amount. The polymer (8) is dissolved in a suitable solvent such as 1.2-dimethoxyethane, diglyme, etc. and then added. The reaction temperature is
Approximately -70 to -80°C is suitable, and the reaction time is approximately 3
0 minutes to 3 hours is appropriate.

Specific examples of the present invention will be explained below. *Metal sodium and liquid ammonia are mixed and kept at -78°C, and a solution of polymer (8) in 1,2-dimethoxyethane is gradually added dropwise. 1,2-dimethoxyethane must be sufficiently dried with metallic sodium in advance.

During the reaction, it is best to stir the system to make it homogeneous. After reacting for a predetermined time, the reaction is stopped by adding ethanol or ammonium chloride, and a small amount of water is added and left overnight to evaporate ammonia. Unreacted polymer is removed by extraction with methylene chloride. After dialysis against distilled water for 3 days and removing any insoluble parts,
The insoluble portion is vacuum-dried, and the soluble portion is concentrated and freeze-dried to obtain the amino sugar polymer (9) of the present invention (wherein, Bn and .n are the same as above.

) Hereinafter, the present invention will be explained with reference to Examples. Example 1 Polymer (8-c) (0.30y) was mixed with 1,2-dimethoxyethane (1,2-dimethoxyethane) which had been previously dried with sodium.
10ml).

This solution was kept at -78°C using liquid ammonia (4
0m1) and metallic sodium (0.34y) 14.8m
m01) under a nitrogen stream. -7 8 reaction system
The mixture was stirred at ℃, and after 2 hours, ethanol was added until the blue color of the reaction system disappeared. Further water (10 ml) is added and the mixture is left at room temperature overnight to evaporate the ammonia. Water (50
m1), washed 5 times with methylene chloride, and washed 3 times with distilled water.
Dialyze for days. The precipitate is separated from P and dried in vacuo, and the solution 1 is concentrated and freeze-dried to obtain the amino sugar polymer (9-c). No difference is observed in the respective IR spectra.
Water soluble part 0.0147y, water insoluble part 0.0955y (total 0.1102y) Yield 83.3%) [Polymer (9
- Physical properties of c)] Ninhydrin reaction: +IR
:3400C77L-1, 1590 (V7l-1 Examples 2-3 The reaction was carried out in the same manner as in Example 1, except that the starting materials azido sugar polymer (8) and the alkali metal were changed.

Claims (1)

[Claims] 1. An amino sugar polymer represented by the following formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (wherein, n represents an integer from 70 to 120). 2 Formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (However, in the formula, Bn represents a benzyl group, and n is 70
represents an integer from 120 to 120. ) is treated with an alkali metal in liquid ammonia to produce an amino sugar represented by the formula: ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (where n in the formula is the same as above). A method for producing an amino sugar polymer, characterized by obtaining a polymer.