JP2751004B2 - Novel polysaccharide derivative, its production method and its use - 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 polysaccharide derivative, a method for producing the same, and a use thereof. More specifically, the present invention relates to a chemical bond between a silane agent and a porous carrier silica gel, and further converts a polysaccharide to the compound at its reducing terminal. The present invention relates to a novel polysaccharide derivative in which a specific substituent is introduced into a part or all of hydroxyl groups of a polysaccharide moiety, a method for producing the same, and a use thereof.

[0002]

2. Description of the Related Art It has been known that a separating agent comprising a compound obtained by physically supporting a polysaccharide such as cellulose or amylose or a derivative thereof on silica gel is useful for optical resolution. I have. However, since this compound has low solvent resistance, there is a drawback that usable eluents are limited when used in liquid chromatography or the like. In addition, the usefulness of the polysaccharide cannot be sufficiently brought out, and the washing solution used for washing the contaminated column is limited, which causes column deterioration.

In order to solve such problems, it has been proposed to use a compound obtained by chemically bonding a polysaccharide derivative to silica gel. However, in this case, since the site to be chemically bonded to the silica gel cannot be selected, the higher order structure of the saccharide itself is affected, and the useful property of the polysaccharide is greatly reduced. In addition, there is a drawback that the quality of the obtained silica gel compound varies greatly due to the unspecified binding site.

The present inventors have already solved the above-mentioned problems by using silica gel as a porous carrier and allowing a polysaccharide to form an amide bond at the reducing end thereof on the inner and outer surfaces of the pore (Japanese Patent Application No. 5-135170). However, Japanese Patent Application No. 5-13
In order to produce the substance described in the specification of US Pat. No. 5,170, the reducing end of the sugar to be bound must be lactonized,
Since it is difficult to lactonize the reducing end in the state of polysaccharide, lactonization is performed at the time of oligosaccharide, and then the sugar chain is extended by enzyme synthesis. For this reason, this method: (1) Unable to synthesize a compound useful as a separating agent when there is no synthetic enzyme for polymerizing a useful oligosaccharide to a polysaccharide. (2) Even if there is a useful enzyme, it is difficult to purify the enzyme. (3) Enzymes and enzyme substrates are expensive and impractical. (4) The reaction process is complicated. And so on.

Therefore, the present inventors have focused on utilizing the polysaccharide as it is, and have found that the carbonyl group at the reducing end of the polysaccharide has
The present inventors have found for the first time a method of synthesizing a target compound by directly chemically bonding to a silica gel surface-treated to have an amino group in the presence of a reducing agent, and have completed the present invention. The compound represented by the above general formula (1), which is a novel substance synthesized by this method, is obtained by chemically bonding only the carbon at the reducing terminal of the sugar moiety to the inner and outer surfaces of the porous carrier via a silane treatment agent. It is.

[0006]

That is, according to the present invention, a polysaccharide or a derivative thereof is chemically bonded to the inner and outer surfaces of pores of silica gel at a reducing end of the polysaccharide site or the derivative site. The present invention provides a novel polysaccharide derivative having a structure represented by the following formula:

[0007]

Embedded image (Where R is Ra,

[0008]

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[0009]

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Wherein Ra is a hydrogen atom or a substituent selected from a substituted or unsubstituted alkyl group, a substituted or unsubstituted phenyl group and a substituted or unsubstituted heterocyclic compound; In order to improve remarkably, the substitution degree of R is preferably 30% or more and 100% or less. Rb is a group in which a substituted or unsubstituted methylene group, a substituted or unsubstituted phenylene group, a hetero atom or the like is covalently bonded. Z represents any one of a porous carrier surface, a halogen atom, an alkyl group, an alkoxy group, a phenyl group, a silane treatment agent and a sugar-bonded silane treatment agent. m ₁ indicates the number of monosaccharide units, indicates an average number of 10 to 500,
m ₂ represents an integer of 1 to 20. )

Further, after the silane treatment agent and the porous carrier are chemically bonded, the chemically bonded compound and the polymerization degree 11
A method for producing a novel polysaccharide derivative represented by the above general formula (1), wherein the polysaccharide or derivative thereof is chemically bonded at the reducing end of the polysaccharide or derivative thereof having a polymerization degree of 11 or more and 500 or less. A chemical bond with the silane treatment agent at the reducing end, and then a chemical bond between the chemically bonded compound and the porous carrier, wherein the novel polysaccharide derivative represented by the general formula (1) is obtained. It is intended to provide a production method and a separating agent for chromatography containing the polysaccharide derivative.

The compound obtained by chemically bonding the silane treatment agent and the porous carrier used in the production of the novel polysaccharide derivative represented by the above general formula (1) is obtained by a known reaction between the silane treatment agent and the silica gel of the porous carrier. It can be obtained by bonding. As a method of chemically bonding a carbon at the reducing end of the polysaccharide to the silane-treating agent, a Schiff base is formed between the aldehyde group and the amino group, and then reduced in the presence of a reducing agent.
How to make a higher amine (Glycoconjugate J (1986) 3, 311
-319 ELISABETH KALLIN et al.) (Hereinafter abbreviated as reductive amination method) can be used. In addition, the following chemical formula (A)
, An amide bond of the following chemical formula (C), a thiourethane bond of the following chemical formula (D), and the like.
It is also possible to bind to the porous carrier with only carbon at the position.

[0013]

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[0014]

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[0015]

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[0016]

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(In the above formula, R, Rb, m ₁ and m ₂ have the same meanings as described above.)

The polysaccharide used in the present invention is a synthetic polysaccharide,
Any of natural polysaccharides and polysaccharide derivatives may be used as long as they have a reducing end and are optically active, but preferably have a high degree of regularity in the binding mode. For example, α-
1,4-glucan (amylose), β-1,4-glucan (cellulose), α-1,6-glucan (dextran), β-1,6-glucan (pustulan), α-1,
3-glucan, β-1,3-glucan (eg, curdlan, schizophyllan), α-1,2-glucan, β-
1,2-glucan, β-1,4-chitosan, β-1,4
-N-acetylchitosan (chitin), β-1,4 galactan, α-1,6-galactan, β-1,2-fructan (inulin), β-2,6-fructan (levan),
β-1,4-xylan, β-1,3-xylan, β-
1,4-mannan, α-1,6-mannan, pullulan,
Agarose, alginic acid, starch having a high amylose content, and more preferably cellulose, amylose, β-1,4-chitosan, chitin, β-1,4-mannan, from which a high-purity polysaccharide can be easily obtained; β-1,4-xylan, inulin, curdlan and the like. The number average degree of polymerization of these polysaccharides is 11 or more, and there is no particular upper limit, but it is preferably 500 or less for ease of handling.

The porous carrier used in the present invention is a porous inorganic carrier or a porous organic carrier, and specifically, silica gel, diatomaceous earth, porous glass, hydroxyapatite, alumina, titanium oxide, magnesia And porous organic carriers such as polyacrylamide and polyacrylate, and silica gel is particularly desirable. Silica gel has a particle size of 1 to 1000 μm, preferably 2 to 100 μm, and an average pore size of 1 nm to 1 μm.
00 μm, preferably 2 nm to 500 nm. In addition, by using a porous membrane as a porous carrier,
It is also possible to obtain a substance useful as a novel separation membrane for chromatography.

When the reductive amination method is used, a silane treating agent having an amino group is used, and a silane treating agent having a primary amine is particularly preferable. The silane treatment agent may be arbitrarily used, such as a commercially available silane coupling agent or a compound synthesized with an amine.

Further, it has a spacer for bonding the silane treatment agent and the sugar, that is, two or more functional groups of the same or different kind, and one of the functional groups chemically bonds to the reducing end of the sugar and the other functional group. And those chemically bonded to the silane treatment agent may be used. Here, the functional group includes, for example, vinyl group, amino group, hydroxy group, carboxyl group, aldehyde group, isocyanate group, thiocyanate group, isothiocyanate group, thiol group, silanol group, epoxy group, ether group, ester group, Examples include an amide group and a halogen atom. The silane treatment agent may be any one capable of binding to these functional groups, and representative silane treatment agents are shown below. Note that Rb in the general formula (1) represents a group in which a substituted or unsubstituted methylene group, a substituted or unsubstituted phenylene group, a hetero atom, or the like is covalently bonded. Or a part of a spacer in which a silane treatment agent is chemically bonded.

[0022]

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[0023]

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[0024]

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[0025]

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[0026]

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[0027]

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[0028]

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[0029]

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In the above formula, n ₁ is an integer of ₁ to 3, R ¹ is a hydrogen atom or an alkyl chain having about 1 to 20 carbon atoms or a derivative thereof, and R ² is an alkyl chain having about 1 to 20 carbon atoms or a derivative thereof. , X represents at least one substituted or unsubstituted alkoxy group having 1 to 10 carbon atoms, a halogen atom (preferably a chlorine atom), a hydroxyl group, a substituted or unsubstituted phenoxy group, and Y represents a halogen atom.

The substituent (R) introduced into a part or all of the hydroxyl group of the polysaccharide represented by the following formula (2) is
The hydroxyl group is modified, and two or more different substituents can be introduced into the hydroxyl group in one molecule of the polysaccharide moiety.
Ra in the substituent (R) is selected from a substituted or unsubstituted alkyl group, a substituted or unsubstituted phenyl group and a substituted or unsubstituted heterocyclic compound, and specifically, methyl, ethyl, propyl Group, butyl group, t-butyl group, phenyl group, methylphenyl group, dimethylphenyl group, ethylphenyl group, diethylphenyl group, trimethylsilylphenyl group, alkoxyphenyl group, dialkoxyphenyl group, halogenated phenyl group, dihalogenated phenyl Group, phenylazophenyl group, naphthalene group, anthracene group, pyridyl group, furyl group and the like.

[0032]

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The introduction of these substituents into the hydroxyl group of the sugar moiety of the compound represented by the general formula (2) can be carried out by a known method.

Hereinafter, a method for synthesizing the novel compound of the present invention represented by the general formula (1) will be exemplified. Production method 1 First, a method for synthesizing the compound represented by the above general formula (2) will be exemplified.

Reaction The binding of the polysaccharide having a reducing end to the surface-treated porous silica gel is exemplified.

The surface treatment of the silica-containing agent having an amino group, specifically 3-aminopropyltriethoxysilane, on the inner and outer surfaces of the pores of the silica gel can be carried out by a conventionally known method. This is added to a solution in which a polysaccharide having a reducing end is dissolved in a solvent such as dimethyl sulfoxide (DMSO), and a reducing agent is further added to reduce the amino group and polysaccharide present on the surface of the silica gel at 50-80 ° C.
The reaction is allowed to proceed for 2 hours, followed by reductive amination. At the time of this reaction, acetic acid may be added to neutralize the pH to 6 to 8 for the reaction. The unreacted polysaccharide is removed by washing with a solvent such as DMSO, acetone and hexane, and dried under reduced pressure to obtain the compound represented by the general formula (2).

At this time, NaBH is used as a reducing agent.
₄ (sodium borohydride), NaBH ₃ CN (sodium cyanoborohydride), borane pyridine complex, borane dimethylamine complex, borane trimethylamine, and other borane compounds. Since the reduction of the carbonyl group takes precedence in a strong reducing agent, a reducing agent having a weak reducing power such as sodium cyanoborohydride is preferable. For reductive amination, it is also possible to chemically bond the sugar reducing terminal to the silane treatment agent via a spacer. That is, it is possible to perform reductive amination at the sugar-reducing end portion with a spacer having an amino group as one of the functional groups, and to chemically bond with the silane treatment agent at the other functional group.

Production Method 2 Reaction An example of the bonding between a polysaccharide having a reducing end and a silane treatment agent is described below. A polysaccharide having a reducing end is dissolved in a solvent such as DMSO, a silane treating agent having an amino group and a reducing agent are added, and the reducing end of the polysaccharide is reacted with the amino group at 50 to 80 ° C. and bonded by a reductive amination method. The following general formula (3)
Is obtained. When using this method,
Since the silane treatment agents may partially polymerize with each other, it is preferable to perform the treatment under anhydrous conditions.

[0039]

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Reaction A combination of the compound represented by the general formula (3) obtained by the above reaction with silica gel is exemplified. The polysaccharide derivative represented by the above general formula (3) obtained by the reaction is dissolved in DMSO or lithium chloride-dimethylacetamide (LiCl-DMA) solution or the like, and further activated by adding pyridine as a catalyst and activating silica gel. Is added to the porous silica gel at the silane portion of the compound represented by the general formula (3) in the same manner as in the conventionally known silane treatment method, whereby the compound represented by the general formula (2) is represented. Obtain the compound. The compound represented by the general formula (3) synthesized by the production method 2 has a relatively unstable silane treatment agent, and therefore the production method 1 is more preferable in terms of handling.

Production method 3 The sugar moiety hydroxyl group of the compound represented by the general formula (2) obtained by the above production method 1 or 2 and, for example, 4-methylphenyl isocyanate; 3,5-dimethylphenyl isocyanate;
5-dichlorophenyl isocyanate; phenyl isocyanate or the like is reacted in an anhydrous DMA / LiCl / pyridine solution or anhydrous DMSO / pyridine to replace all or a part of the hydroxyl group in the sugar moiety of the compound, and the compound is represented by the general formula (2). The compounds represented can be obtained. This reaction can be carried out in a known manner.

The amount of the compound chemically reacted with the silica gel is not particularly limited, but is usually preferably about 5 to 50% by weight. In order to eliminate the influence of the residual silanol group of the obtained present compound, the performance as a separating agent can be improved by performing an endcapping treatment by a conventionally known method. In the present invention, in showing the structure of each compound, the structure showing the position of the hydroxyl group of the saccharide and the like is partially simplified for convenience and convenience.

EXAMPLES Hereinafter, the present invention will be described with reference to Examples. Example 1 (1-1) Synthesis of surface-treated silica gel (Part 1) Silica gel (manufactured by Fuji Silysia Chemical Ltd., having an average pore diameter of 50 nm, which was previously activated (vacuum dried at 180 ° C. for 2 hours)
12 ml of anhydrous benzene and 1 ml of anhydrous pyridine were added to 10 g of 10 μm (particle size: 5 μm), 0.7 ml of 3- (2-aminoethylaminopropyl) trimethoxysilane was added, and the mixture was reacted at 90 ° C. for 12 hours. This surface-treated silica gel is washed with methanol, acetone, and hexane, and then
Vacuum dried for a period of time to obtain surface-treated silica gel (1-1 silica gel).

(1-2) Synthesis of Surface-treated Silica Gel (Part 2) Silica gel (manufactured by Fuji Silysia Chemical Ltd., average pore size: 50 nm, previously activated (180 ° C., vacuum drying for 2 hours)
12 ml of anhydrous benzene and 1 ml of anhydrous pyridine were added to 10 g of 10 μm (particle size: 5 μm), 0.7 ml of 3-aminopropyltriethoxysilane was added, and the mixture was reacted at 90 ° C. for 12 hours.
The surface-treated silica gel is washed with methanol, acetone, and hexane, and then dried under vacuum at 60 ° C. for 2 hours to obtain a surface-treated silica gel (1-2 silica gel).

(1-3) Synthesis of compound represented by the following general formula (4)

[0046]

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(Wherein Rc is a hydrogen atom or

[0048]

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And the substituent is 9% by weight analysis.
0% or more. m ₃ is an average of about 100, and Z ₁ is a silica gel surface, a methoxy group and either a silane treatment agent or a sugar-bonded silane treatment agent)

Amylose (1.0 g, average Dp) was added to the surface-treated silica gel (1-1 silica gel) having an aminoethylaminopropyl group bonded to the surface obtained in (1-1) above.
100) dissolved in 8 ml of anhydrous DMSO,
Disperse. Further, a solution prepared by dissolving 150 mg of NaBH ₃ CN in ₅ ml of anhydrous DMSO and a solution obtained by adding 30 mg of acetic acid were added thereto, and the mixture was reacted at 50 ° C. for 12 hours under nitrogen to chemically bond the amino group of the surface-treated silica gel with the reducing terminal of amylose.

The sugar-bonded silica gel thus obtained was filtered using a glass filter G4, and the residue was treated with DMS.
It was washed with O, tetrahydrofuran, methanol, acetone and hexane to remove unbound amylose and the like, and dried under vacuum at 60 ° C. for 2 hours. Elemental analysis (Table 1) confirmed the binding of the sugar to give a compound represented by the following general formula (5). The elemental analysis values were as follows: C: 4.40%, H: 1.03%,
N: 0.40%.

[0052]

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The sugar-bonded silica gel was treated with anhydrous DMA
-Dispersed in a liquid composed of 8 ml of LiCl and 3 ml of pyridine, 3.0 ml of 3,5-dimethylphenyl isocyanate was added to the reaction solution, and reacted at 80 ° C. for 12 hours under nitrogen to chemically bond to the silica gel surface. The hydroxyl group of the sugar moiety was modified. After confirming that an excess of isocyanate group remained in the solution by confirming stretching vibration between C and N at 2270 cm ^-1 in the IR spectrum, the surface-treated silica gel in the reaction solution was treated with tetrahydrofuran, methanol, After washing with acetone and hexane to remove impurities, vacuum drying was performed at 60 ° C. for 2 hours. Then
This compound was subjected to IR spectrum analysis and elemental analysis (Table 1).

[0054]

[Table 1]

As a result, in the IR spectrum, 173
A stretching vibration of a carbonyl group (absorption of C ＝ O of the secondary carbamate) was observed at around 0 cm ⁻¹ , and further, a bond to silica gel was confirmed from an elemental analysis value, and the above formula (4) was used. The compound represented by was obtained. This was used as a separating agent for chromatography. Table 2 shows the results of evaluating this compound as a separating agent for various racemic compounds.

[0056]

[Table 2]

In Table 2, k ₁ is a holding capacity ratio.
Α is a separation coefficient, which is obtained by the following equation. Also, k ₂ is k ₁ and a retention capacitance ratio similarly, those obtained by the following equation.

[0058]

(Equation 1)

[0059]

(Equation 2)

[0060]

(Equation 3)

(1-4) Synthesis of compound represented by the following general formula (6)

[0062]

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(Wherein Rc is a hydrogen atom or

[0064]

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The degree of substitution was 90% based on the weight calculation.
That's it. m ₄ is an average of about 160, and Z ₂ is a silica gel surface, an ethoxy group and either a silane treatment agent or a sugar-bonded silane treatment agent. )

A solution obtained by dissolving 1.0 g of amylose (average Dp160) in 8 ml of anhydrous DMSO was added to the surface-treated silica gel (1-2 silica gel) having an aminopropyl group bonded to the surface obtained in the above (1-2). Disperse.
Further, 150 mg of NaBH ₃ CN was added to ₅ m of anhydrous DMSO.
1 and 30 mg of acetic acid were added, and the mixture was reacted at 50 ° C. for 12 hours under nitrogen to chemically bond the amino group of the surface-treated silica gel with the reducing terminal of amylose.

The sugar-bound silica gel thus obtained was filtered using a glass filter G4, and the residue was treated with DMS.
It was washed with O, tetrahydrofuran, methanol, acetone and hexane to remove unbound amylose and the like, and dried under vacuum at 60 ° C. for 2 hours. Elemental analysis (Table 1) confirmed the binding of the saccharide to give a compound represented by the following general formula (7). The elemental analysis values were as follows: C: 4.29%, H: 0.90%,
N: 0.22%.

[0068]

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This sugar-bonded silica gel was treated with anhydrous DMA-
Disperse in a solution consisting of 8 ml of LiCl and 3 ml of pyridine, add 3.0 ml of 3,5-dimethylphenyl isocyanate to this reaction solution, react at 80 ° C. for 12 hours under nitrogen, and react with the sugar chemically bonded to the silica gel surface. Some of the hydroxyl groups were modified. Excess isocyanate groups remaining in the solution were confirmed by IR spectrum at 2270 cm ^-1.
After confirming by confirming the stretching vibration between C = N,
The surface-treated silica gel in the reaction solution was washed with tetrahydrofuran, methanol, acetone, and hexane to remove impurities, and then dried under vacuum at 60 ° C. for 2 hours. Next, the compound was subjected to IR spectrum analysis and elemental analysis (Table 1).

As a result, 173 was found in the IR spectrum.
The stretching vibration of the carbonyl group (absorption of C ＝ O of the secondary carbamate) was observed at around 0 cm ⁻¹ , and further, the bond to silica gel was confirmed from the elemental analysis values, and the above formula (6) was used. The compound represented by was obtained. This was used as a separating agent for chromatography. Table 2 shows the results of evaluating this compound as a separating agent for various racemic compounds.

(1-5) Synthesis of compound represented by the following general formula (8)

[0072]

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(Wherein Rc is a hydrogen atom or

[0074]

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The substitution degree was 90% based on the weight calculation.
That's it. m ₅ is about 200, Z ₂ is the silica gel surface,
An ethoxy group and either a silane treatment agent or a sugar-bonded silane treatment agent. )

To the surface-treated silica gel (1-2 silica gel) having an aminopropyl group bonded to the surface obtained in the above (1-2), 1.0 g of cellulose (manufactured by Merck) was added in anhydrous D.
A solution dissolved in 21 ml of MA / LiCl is added and dispersed. Further, 150 mg of NaBH ₃ CN was added to anhydrous DM.
A / LiCl dissolved in 5 ml and acetic acid 30 mg
Was added thereto, and the mixture was reacted at 50 ° C. for 36 hours under nitrogen to chemically bond the amino group of the surface-treated silica gel with the reducing terminal of cellulose.

The sugar-bonded silica gel thus obtained was filtered using a glass filter G4, and the residue was treated with DMA.
/ LiCl, tetrahydrofuran, methanol, acetone, hexane to remove unbound amylose, etc.
Vacuum dried at 60 ° C. for 2 hours. Elemental analysis (Table 1) confirmed the binding of the saccharide to give a compound represented by the following general formula (9). The elemental analysis values were as follows: C: 2.12%, H: 0.5
6%, N: 0.15%.

[0078]

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The sugar-bonded silica gel was treated with anhydrous DMA-
Disperse in a solution consisting of 8 ml of LiCl and 3 ml of pyridine, add 3.0 ml of 3,5-dimethylphenyl isocyanate to this reaction solution, react at 80 ° C. for 12 hours under nitrogen, and react with the sugar chemically bonded to the silica gel surface. Some of the hydroxyl groups were modified. Excess isocyanate groups remaining in the solution were confirmed by IR spectrum at 2270 cm ^-1.
After confirming by confirming the stretching vibration between C = N,
The surface-treated silica gel in the reaction solution was washed with tetrahydrofuran, methanol, acetone, and hexane to remove impurities, and then dried under vacuum at 60 ° C. for 2 hours. Next, the compound was subjected to IR spectrum analysis and elemental analysis (Table 1).

As a result, in the IR spectrum, 173
The stretching vibration of the carbonyl group (absorption of C ＝ O of the secondary carbamate) was observed around 0 cm ⁻¹ , and further, the bond to silica gel was confirmed from the elemental analysis value, and the above-mentioned general formula (8) was obtained. The compound represented by was obtained. This was used as a separating agent for chromatography. Table 3 shows the results of evaluating this compound as a separating agent for various racemic compounds.

[0081]

[Table 3]

(1-6) Synthesis of compound represented by the following general formula (10)

[0083]

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(Wherein Rc is a hydrogen atom or

[0085]

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The degree of substitution is 90% from the weight calculation.
That is all. m ₄ is an average of about 160, and Z ₂ is a silica gel surface, an ethoxy group and either a silane treatment agent or a sugar-bonded silane treatment agent. )

To 3 g of the surface-treated silica gel (1-2 silica gel) having an aminopropyl group bonded to the surface obtained in (1-2), 1.0 g of amylose (average Dp160)
Was dissolved in 8 ml of anhydrous DMSO and dispersed. Further, 150 mg of NaBH ₃ CN was added to anhydrous DM.
A solution dissolved in 5 ml of SO and a solution to which 30 mg of acetic acid was added were added, and the mixture was reacted at 50 ° C. for 12 hours under nitrogen to chemically bond the amino group of the surface-treated silica gel with the reducing terminal of amylose.

The sugar-bound silica gel thus obtained was filtered using a glass filter G4, and the residue was treated with DMS.
It was washed with O, tetrahydrofuran, methanol, acetone and hexane to remove unbound amylose and the like, and dried under vacuum at 60 ° C. for 2 hours.

The sugar-bonded silica gel was treated with anhydrous DMA
-Dispersed in a liquid composed of 8 ml of LiCl and 3 ml of pyridine, and dissolved in 5 ml of DMA in this reaction solution.
-Dichlorophenyl isocyanate (3,5
5 g was added, and the mixture was added under nitrogen under 8 g.
The reaction was carried out at 0 ° C. for 12 hours to modify the hydroxyl group of the sugar moiety chemically bonded to the silica gel surface. After confirming that an excess isocyanate group remained in the solution by confirming stretching vibration between C and N at 2270 cm ^-1 in the IR spectrum, the surface-treated silica gel in the reaction solution was treated with tetrahydrofuran, methanol, After washing with acetone and hexane to remove impurities, vacuum drying was performed at 60 ° C. for 2 hours. Next, the compound was subjected to IR spectrum analysis and elemental analysis (Table 1).

As a result, in the IR spectrum, 173
The stretching vibration of the carbonyl group (absorption of C ＝ O of the secondary carbamic acid ester) was observed around 0 cm ⁻¹ , and further, the bond to silica gel was confirmed from the elemental analysis value, and the above formula (10) was obtained. The compound represented by was obtained. This was used as a separating agent for chromatography. Table 2 shows the results of evaluating this compound as a separating agent for various racemic compounds.

(1-7) Synthesis of compound represented by the following general formula (11)

[0092]

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(Wherein Rc is a hydrogen atom or

[0094]

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The degree of substitution was 90% based on the weight calculation.
That is all. m ₄ is an average of about 160, and Z ₂ is a silica gel surface, an ethoxy group and either a silane treatment agent or a sugar-bonded silane treatment agent. )

To 3 g of the surface-treated silica gel (1-2 silica gel) having an aminopropyl group bonded to the surface obtained in the above (1-2) was added 1.0 g of amylose (average Dp 160).
Was dissolved in 8 ml of anhydrous DMSO and dispersed. Further, 150 mg of NaBH ₃ CN was added to anhydrous DM.
A solution dissolved in 5 ml of SO and a solution to which 30 mg of acetic acid was added were added, and the mixture was reacted at 50 ° C. for 12 hours under nitrogen to chemically bond the amino group of the surface-treated silica gel with the reducing terminal of amylose.

The sugar-bound silica gel thus obtained was filtered using a glass filter G4, and the residue was treated with DMS.
It was washed with O, tetrahydrofuran, methanol, acetone and hexane to remove unbound amylose and the like, and dried under vacuum at 60 ° C. for 2 hours.

This sugar-bonded silica gel was treated with anhydrous DMA-
Disperse in a liquid composed of 8 ml of LiCl and 3 ml of pyridine, and add 2 ml of phenyl isocyanate to the reaction solution.
Was added thereto and reacted at 80 ° C. for 12 hours under nitrogen to modify the hydroxyl group of the sugar moiety chemically bonded to the silica gel surface. The presence of excess isocyanate groups in the solution was confirmed by IR
After confirming that the stretching vibration between C and N was observed at 2270 cm ^-1 in the spectrum, the surface-treated silica gel in the reaction solution was washed with tetrahydrofuran, methanol, acetone and hexane to remove impurities.
Vacuum dried at 2 ° C. for 2 hours. Next, the compound was subjected to IR spectrum analysis and elemental analysis (Table 1).

As a result, 173 in the IR spectrum
The stretching vibration of the carbonyl group (absorption of C ＝ O of the secondary carbamate) was observed around 0 cm ⁻¹ , and further, the bond to silica gel was confirmed from the elemental analysis value, and the above formula (11) was obtained. The compound represented by was obtained. This was used as a separating agent for chromatography. Table 2 shows the results of evaluating this compound as a separating agent for various racemic compounds.

Application Example Preparation of Optical Resolution Column and Optical Resolution A novel compound obtained in Example 1 (1-3) was obtained in an amount of 0.46 × 2.
An empty 5 cm stainless steel column was packed by a slurry filling method. The filling apparatus used was a PS10, PS-20 auto-packing system manufactured by Kyoto Chromato. Using this column, the optical resolution of the compound was evaluated. The high performance liquid chromatograph system used a Waters 510 pump and a 486 UV detector. Further, as a comparative example, a product prepared by lactonizing amylose and chemically bonding to surface-treated silica gel through an amide bond was used as a separating agent.

Further, in order to examine the solvent resistance of the column for optical resolution obtained by the present invention, the optical resolution was examined after flowing a tetrahydrofuran (THF) solution at a flow rate of 1 ml / min for 2 hours. Was not recognized, and excellent solvent resistance was confirmed. The analysis was performed using eluent → hexane:
IPA = 90: 10, # indicates 95: 5. Also, the flow velocity
The test was performed under the conditions of 0.5 ml / min and room temperature.

[0102]

The novel substance of the present invention has excellent solvent resistance and is useful as a chromatographic separating agent for resolving optically active substances. Further, according to the present invention, the novel substance can be efficiently produced.

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Hirofumi Akano 2-46-28 Ariwaki-cho, Handa-shi, Aichi Prefecture

Claims (4)

(57) [Claims] 1. A polysaccharide or a derivative thereof is chemically bonded to the inner and outer surfaces of pores of a porous carrier at a reducing end of the polysaccharide site or the derivative site, and mainly has a structure represented by the following general formula (1). A novel polysaccharide derivative having a structure. Embedded image (Wherein R is Ra, Embedded image Wherein Ra is a hydrogen atom or a substituent selected from a substituted or unsubstituted alkyl group, a substituted or unsubstituted phenyl group and a substituted or unsubstituted heterocyclic compound, and Rb is a substituted or unsubstituted A compound in which a methylene group, a substituted or unsubstituted phenylene group, a hetero atom or the like is covalently bonded. Z represents any one of a porous carrier surface, a halogen atom, an alkyl group, an alkoxy group, a phenyl group, a silane treatment agent and a sugar-bonded silane treatment agent. m ₁ indicates the number of monosaccharide units, and on average 10 to 500
And m ₂ represents an integer of 1 to 20. ) 2. After chemically bonding a silane treatment agent and a porous carrier, the chemically bonded compound and a polymerization degree of at least 11
2. The method for producing a novel polysaccharide derivative according to claim 1, wherein a chemical bond is made at the reducing end of the polysaccharide or its derivative of 00 or less. 3. The method according to claim 1, wherein the reducing end of the polysaccharide having a degree of polymerization of 11 or more and 500 or less is chemically bonded to a silane treatment agent, and further chemically bonded to the porous carrier. A method for producing a novel polysaccharide derivative according to claim 1, wherein 4. A separating agent for chromatography containing the novel substance according to claim 1.