JPH0368602A - Cyclodextrin having formylated branch and preparation thereof - Google Patents

Abstract

PURPOSE:To enable the formylation of only the branched part to thereby enable immobilization by reacting a branched cyclodextrin with a guest compd. and oxidizing the reaction product with periodic acid. CONSTITUTION:One mol of a branched cyclodextrin having a branched part comprising a saccharide contg. at least two vicinal OH groups and a cyclic part comprising alpha-, beta-, or gamma-cyclodextrin is mixed with 0.1-0.3mol of a guest compd. and is allowed to stand for 1-24hr at room temp. to cause the branched cyclodextrin to include the guest compd. 0.001-1mol of a periodata is added to the reaction product, which is then treated in an aq. solvent at 0-20 deg.C for 0.5-10hr and, if necessary, washed and purified to give a branched cyclodextrin wherein only the branched part is formylated.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a branched cyclodextrin in which only the branch portions are aldehyded, and a method for producing the same.

[Conventional technology]

Cyclodextrin is a compound obtained by treating starch with cyclodextrin glucanotransferase and has a cyclic structure in which 6 to F2 glucose residues are bonded with α-1,4. Common cyclodextrins that are industrially mass-produced include α-cyclodextrin, which is composed of 6 glucose residues, and 7
There are three kinds of β-cyclodextrin, 8 β-cyclodextrin and 8 T-cyclodextrin. Since cyclodextrin has the function of including various molecules in its cylindrical structure, various uses have been developed that utilize this function.

Various studies have also been conducted on cyclodextrin derivatives. For example, Deshiro et al. attempted to improve the catalytic effect by combining metal complexes with β-cyclodextrin modified with ethylenediamine as ligands [J, Am, C
hem, Sac, 99.7100 (1977)
]. In addition, Kikuchi et al. formed a complex between methylated cyclodextrin and 1-hexylcarbamoyl-5-fluorouracil, a type of anticancer drug, and investigated the solubility, stability, and stability of the complex as part of their investigation of the antitumor effect. in
It has been reported that in vivo absorption is improved [
[ncl, Phenom, .

2.623 (1984) and Int, J., Phar.
m.

38.191 (1987)). And Ueno et al.
We are considering the inclusion of guest compounds by attaching naphthyls to the primary hydroxyl group area of cyclodextrin [J
, Chem, Sac,, Chem, Commu
n.

(1983)]. Furthermore, Yamaguchi et al. succeeded in isolating and purifying IJn (Etherified cyclodextrin) [Special Publication No. 1]
-24401].

As described above, research and development regarding cyclodextrin is actively conducted, but there is a problem with the solubility of cyclodextrin itself. In this regard, Kobayashi and Kainuma et al. have established a method for efficiently producing glucosyl cyclodextrin, and have obtained branched cyclodextrin (IJ) with excellent solubility [Starch Science, 30.231 (19
83)].

As described above, although the solubility has been greatly improved by producing branched cyclodextrins, there is basically no difference in their uses and usage from conventional cyclodextrins. In other words, as in the case of cyclodextrin + J, the target substance is included in the cavities of branched cyclodextrin in an aqueous solution, powdered through a process such as drying or concentration, and the included target substance is recovered and used. The method is common. However, it is difficult to handle and use the branched cyclodextrin containing the target substance in an aqueous solution by separating it outside the system and is expensive.

Therefore, a technology to immobilize cyclodextrin has been developed, making it possible to recover and separate target substances from aqueous solutions. For example, a method of cross-linking cyclodextrin with a cross-linking agent such as epichlorohydrin to form a bead-like polymer to adsorb and reduce bitter components in grapefruit juice and navel juice (J, Agric, Food
Chem, 32.832836 (1984)], cyclodextrin immobilized on an acrylic resin carrier via a spacer, cyclodextrin crosslinked using an acrylic resin as a crosslinking agent, and a spacer attached to a silica carrier. There are also those in which cyclodextrin is immobilized via a cyclodextrin.

However, when cyclodextrin is immobilized via a spacer, there is a problem in terms of cost when used industrially in large quantities. Furthermore, when a bead-shaped cyclodextrin polymer crosslinked with an acrylic resin is used for adsorption and fractionation of useful components in an aqueous solution, it takes time to recover the components adsorbed at the center of the beads. Furthermore, cyclodextrin polymers obtained with crosslinking agents such as epichlorohydrin have problems in terms of safety.

By the way, derivatives related to the above-mentioned branched cyclodextrin, especially derivatives having a functional group only at the branch portion, have not been obtained. Therefore, immobilization of this branched cyclodextrin has not yet been performed.

[Problem to be solved by the invention]

It is therefore an object of the present invention to provide a branched cyclodextrin with useful functional groups for immobilization and a method for producing the same.

[Means to solve the problem]

The present invention provides an aldehyded branched cyclodextrin in which only the branch portion of the branched cyclodextrin is aldehyded.

The present invention also provides a method for producing an aldehyded branched cyclodextrin in which only the branched portion is aldehyded, which is characterized in that a guest compound is included in the branched cyclodextrin and then subjected to periodic acid oxidation treatment. This is what we provide.

The branched cyclodextrin ft used in the present invention is a saccharide in which the branching part has at least two adjacent ○H groups, such as glucosyl, maltosyl, and vanosyl, and the cyclic part is α-1β- and T-cyclodextrin. It is preferable to use one of the following. These may be monobranched cyclodextrins, homologous polybranched cyclodextrins, or heterologous polybranched cyclodextrins.

To produce the aldehyded branched cyclodextrin of the present invention, a suitable guest compound is first included in the branched cyclodextrin. Examples of such guest compounds include butanol, cyclohexane, cyclohexanol, toluene, and phenols (phenol,
2-Nitrofene/-JL, 3-Nitrophe/-JLt
, 4-nitrophenol, 3-cyanophenol, 4-
Cyanophenol, 3,5-dimethylphenol, p-
nitrophenol, etc.), benzoic acids (benzoic acid, 〇-aminobenzoic acid, m-aminobenzoic acid, p-aminobenzoic acid, 0-nitrobenzoic acid, m-nitrobenzoic acid, p-nitrobenzoic acid, 2-methyl Benzoic acid, 3-methylbenzoic acid, 4-methylbenzoic acid, 0-hydroxybenzoic acid, m
-hydroxybenzoic acid, p-hydroxybenzoic acid, etc.),
Alcohols (ethanol, 2-methyl-2-propanol, 212-dimethyl-1-propanol, 1-butanol, 1-pentanol, l-hexanol, etc.), organic acids (formic acid, acetic acid, propionic acid, 3-ethylphenylacetic acid, Trimethylacetic acid, etc.>, amino acids (L-phenylalanine, L-tyrosine, L-tryptophan, etc.), pyridine, indole, arylisothiocyanate, 1.
Examples include 4-dioxane and acetonitrile. Guest compound is 0.0% per mole of branched cyclodextrin.
It is preferable to use about 1 to 0.3 mol. Mix branched cyclodextrin (branched cyclodextrin) IJ and a guest compound,
When left for 24 hours, the guest compound is included.

After removing excess guest compound as necessary, this is subjected to periodic acid oxidation treatment. Add a periodate salt such as periodic acid or sodium metaperiodate to give a concentration of 0.00 to 1 to 1 mole, and add 0.5 to 10 moles of periodate at 0 to 20°C in an aqueous solvent.
By time treatment, branched cyclodextrin) IJ
Only the branch of the chain is aldehyded. 2 aldehyde groups when using glucosylcyclodextrin, 2 or 4 aldehyde groups when using maltosylcyclodextrin, 2 when using panosylcyclodextrin , 4 or 6 aldehyde groups are formed.

Next, excess periodic acid is decomposed by adding ethylene glycol, etc., and then adsorbed onto an activated carbon column and washed with water. After washing with water, it is eluted with 15-50% ethanol, and purified by concentration and recrystallization.

〔Effect of the invention〕

The aldehyde-branched cyclodextrin of the present invention can be easily immobilized by chemical bonding to a carrier having an amino group, such as an amino group-containing polysaccharide, protein, or amino group-containing synthetic resin. The cyclodextrin immobilized in this manner can be packed into a column in the same way as ion exchange resins and the like, and can be advantageously used for the removal and recovery of specific useful substances and harmful substances.

Example 1 1.4 g of glucosyl β-cyclodextrin and 16 rn of cyclohexane! ! was added and left overnight to form a clathrate. Excess cyclohexane was removed with a Pasteur pipette, 950 ml of distilled water was added thereto, and the mixture was stirred well.
50ml of 1M aqueous sodium metaperiodate solution! were added, mixed, and left to react at 5° C. for 3 hours while shielding from light. Of the 5 mmol periodic acid, 2 mmol was consumed. As a control experiment, a similar reaction was carried out using β-cyclodextrin without side chains, but periodic acid was not consumed. Next, add ethylene glycol 2- to give 15
The excess meta-periodic acid was left to stand for a minute to decompose the IJium.

Next, the reaction solution was passed through an activated carbon column in a dark place, and after thorough washing with water, 30 to 50% ethanol was passed through the column, and the eluate was concentrated to dryness. A small amount of distilled water was added to dissolve it, 50% of ethanol was added to recrystallize it, and S was condensed to dryness to obtain β-cyclodextrin with an aldehyde group in the side chain.
．． 9g was obtained.

Example 2 T-cyclodextrin 20g and maltose 150g
g of 50 mmol acetate buffer (ρ113.75) 175
ml was used as a substrate to react with the immobilized pullulanase enzyme at 50 to 52°C for 5 to 7 days. The reaction solution was passed through an activated carbon column, and unreacted maltose was removed by washing with water and 7.5-15% ethanol. (2) Elution was carried out with 5 to 50% ethanol, and the eluate was concentrated to dryness, redissolved in a small amount of distilled water, and recrystallized by adding ethanol. After vacuum drying, 7.8 g of maltosyl T-cyclodextrin was obtained.

20 ml of cyclohexanol was added to 5 g of maltosyl γ-cyclodextrin thus obtained, and
The mixture was shaken at ℃ to include cyclohexanol. After inclusion, 50 mmol acetate buffer (+) H4,5) 10
00-, glucoamylase (Glucozyme NL-3 manufactured by Amano Pharmaceutical Co., Ltd.) Ig was added, and the mixture was reacted at 40°C for 2 hours. After the reaction, glucoamylase was inactivated by boiling at 100° C. for 5 minutes, the reaction solution was passed through an activated carbon column, and unreacted maltose was removed by washing with water. After washing thoroughly with water 3
Elution was performed with 0-50% ethanol, and the eluate was concentrated to dryness. After dissolving by adding a small amount of distilled water, ethanol was added for recrystallization, and 2.65 g of glucosyl γ-cyclodextrin was obtained by concentration to dryness.

Next, glucosylarcyclodextrin 2.65 g
Toluene 40- was added to the annular part to include toluene, and then dissolved in distilled water of i o o oil'. 0,
A 1 molar sodium metaperiodate solution (100 -) was added, mixed well, and reacted for 4 hours in a refrigerator at 4°C. Ethylene glycol was added and the mixture was allowed to stand for 15 minutes to decompose excess periodic acid, followed by purification with an activated carbon column to obtain 1.8 g of γ-cyclodextrin having an aldehyde group in the side chain.

Reference Example 1 1.0 g of β-cyclodextrin having an aldehyde group in the side chain obtained in Example 1 was dissolved in 400 g of distilled water, 10 g of partially deacetylated chitin was added, and the mixture was magnetically heated for 3 hours. The mixture was stirred with a stirrer to react. By centrifugation, 10.5 g of chitin with β-cyclodextrin immobilized on its surface was obtained. The weight content of cyclodextrin on the chitin surface of the main island is 5%.
Met.

After acetylation treatment to eliminate excess amino groups on the main island, 10 g of crystal was packed in a column and 0.1% Naringin 30- was passed through it. Approximately 29% of benaringin was removed compared to chitin (deacetylated chitin).

The amount of naringin was determined by colorimetrically fixing the glucose produced by the action of naringinase (at 40° C. for 20 minutes) using the glucose oxidase method.

Reference Example 2 3 g of T-cyclodextrin having an aldehyde group in the side chain obtained by periodic acid oxidation of the branched portion of the glucosyl T-cyclodextrin obtained in Example 2 was added to 500 ml of distilled water! It was dissolved in

Add to this the weakly basic anion exchange resin Amberlite IRA.
30g of -45 was added and reacted for 3 hours while stirring with a stirrer.

Weakly basic anion exchange resin Amberlite IRA- on which branched T-cyclodextrin is immobilized by filtration.
45 (30.6 g) was obtained. When the amount of aldehyded γ-cyclodextrin in the supernatant before and after the reaction was measured by the phenol-sulfuric acid method, it was found to have decreased by 25% after the reaction. Therefore, it can be seen that 25% of the aldehyded T-cyclodextrin used is bound to the resin.

In addition, the total amine content of Honjima and weakly basic anion exchange resin Amberlite IRA-45 was 51 gg1a.

Confirmed using the Hanna and Kervenski method, the total number of amine moles/sample (g) on the main island is 1.91X10
-', total amine moles of control/sample (g) is 2.79
It was X1O-4. Therefore, it was confirmed that the amino groups of the weakly basic anion exchange resin T Inverlite IRA-45 were reduced by the Schiff base reaction.

Table 1 shows the physical properties of cyclodextrin having two aldehyde groups in its side chain, which is obtained by converting glucosyl cyclodextrin into aldehyde.

[Brief explanation of drawings]

Figure 1 shows β-cyclodextrin (β-cyclodextrin) in which the glucosyl moiety of glucosyl β-cyclodextrin has been converted into a dialdehyde.
Figure 2 shows the infrared absorption spectrum of γ-cyclodextrin (γ-cyclodextrin), in which the glucosyl moiety of glucosyl-arcyclodextrin has been converted into a dialdehyde. Flat bottom 21 Title of invention Aldehyde-branched cyclodextrin and process for producing the same 3. Person making the amendment Relationship with the case 4. 5. Date of amendment order The following sentence is added at the end of page 17 of the specification. "Figure 3 shows the infrared absorption spectrum of α-cyclodextrin in which the glucosyl moiety of glucosyl α-cyclodextrin has been converted into dialdehyde." Figure 3 is added as an attachment.

Claims (2)

[Claims] (1) Aldehydated branched cyclodextrin in which only the branched portion of the branched cyclodextrin is aldehyded. (2) A method for producing an aldehyded branched cyclodextrin in which only the branched portion is aldehyded, which comprises including a guest compound in the branched cyclodextrin and then subjecting it to periodic acid oxidation treatment.