JPH0446961B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides a novel cyclodextrin polymer resin, more specifically, a polycyclodextrin resin having the general formula NH ₂ -(CH ₂ ) _o -NH- (in which n is 2 to 20). This invention relates to a method for producing a polycyclodextrin resin obtained by introducing an aminoalkylamino group represented by the following formula.

Cyclodextrin is a cyclic oligosaccharide in which six or more glucose units are α-1,4-linked,
It is a substance that has a cylindrical cavity within its molecule and can envelop various substances in the cavity to form clathrate compounds. Substances that have such clathrate-forming properties form clathrate compounds by incorporating into them substances with molecular shapes and sizes that suit the cavity, and depending on the type of clathrate substance, Forms clathrate compounds with different affinities. By utilizing these properties of cyclodextrin, it is possible to divide or isolate a specific compound or group of compounds from a mixture of various substances. When used for practical purposes such as substance separation, cyclodextrin is usually made insolubilized by, for example, cross-linked polymerization, and then used by filling it into a chromatography column.

Incidentally, cyclodextrin can be made into a gel-like insoluble resin by polymerizing it using a crosslinking agent such as epichlorohydrin or a diepoxy compound, but such resins only exhibit the same clathration effect as cyclodextrin. Rather, it has properties similar to those of polysaccharide gels such as dextran gels. In the case of cyclodextrin resins, attempts have been made to utilize the characteristics of cyclodextrin, but no attempts have been made yet to utilize characteristics common to other polysaccharide gels. Considering that cyclodextrin production technology has developed in recent years and highly purified products are now available at low cost, the development of more general-purpose usage methods is also considered promising.

As a result of extensive research into various uses of polycyclodextrin resin obtained by cross-linking polymerization of cyclodextrin, the present inventors found that polycyclodextrin resin has the general formula NH ₂ −(CH ₂ ) _o −
We have discovered that by introducing an aminoalkylamino group represented by NH-, a polycyclodextrin resin that can be used as a carrier for enzyme immobilization and a carrier for affinity chromatography can be obtained, and we have accomplished the present invention. Ivy.

That is, in the present invention, a sulfonyl chloride represented by the general formula R-SO ₂ Cl (R in the formula is an alkyl group or _an aryl group) is applied to a polycyclodextrin resin, and then a sulfonyl chloride represented by the _{general formula} −
Obtained by reacting a diaminoalkane represented by NH ₂ (n in the formula is a number from 2 to 20),
The present invention provides a polycyclodextrin resin having an aminoalkylamino group and a method for producing the same.

The cyclodextrin used to form the polycyclodextrin resin in the present invention is α-
Cyclodextrin, β-cyclodextrin,
or γ-cyclodextrin, or a combination of two or three of these may be used. Polycyclodextrin resin is a polymerized version of these cyclodextrins, and any resin can be used as long as it can withstand the reaction of introducing aminoalkylamino groups and sulfonyl groups. A crosslinked polymerized polycyclodextrin resin is preferred. Such cross-linked polymerized resins have a structure as shown in Figure 1, and are extremely resistant to heat and alkali, as they also have a large number of hydroxyl groups to react with sulfonyl chloride. suitable.

In the method of the present invention, such an insoluble polycyclodextrin resin is swollen in a solvent and then reacted with sulfonyl chloride. Examples of the sulfonyl chloride include benzenesulfonyl chloride, p-toluenesulfonyl chloride, and naphthalenesulfonyl chloride. Aromatic sulfonyl chlorides such as chloride and alkylsulfonyl chlorides are used.

In this reaction, the polycyclodextrin resin is vacuum dried at 60° C. to 100° C. using phosphorus pentoxide, dispersed in 5 to 50 times the weight of dehydrated pyridine, and then sulfonyl chloride is added.
Since solid polycyclodextrin resin swells in pyridine and the mechanical strength of the resin decreases, the polycyclodextrin resin is preferably used in the reaction in the form of beads, but it can also be used in the form of pulverized powder.

The amount of sulfonyl chloride added to the dispersion is selected depending on the desired rate of introduction of sulfonyl groups, but it is usually preferably in the range of 0.1 to 1 part by weight per 1 part by weight of the polycyclodextrin resin.
However, if an extremely high or extremely low introduction rate of sulfonyl groups is desired, an amount outside this range can be added.

Sulfonyl chloride is added and dissolved in a pyridine solution in which insoluble polycyclodextrin resin is swollen and dispersed, and the sulfonyl chloride is kept at room temperature to 50°C and allowed to react for 1 hour to several days. A polycyclodextrin resin having the following properties is obtained.

PCD-OH+R- _SO2Cl → PCD-O- _SO2 -R+HCl Here, PCD indicates polycyclodextrin. After the reaction is completed, the reaction product is separated by filtration or the like, thoroughly washed with acetone or lower alcohol, and then dried.

The thus obtained polycyclodextrin resin having a sulfonyl group is reacted with a diaminoalkane to introduce an aminoalkylamino group according to the following reaction.

PCD−O−SO ₂ R+NH ₂ −(CH ₂ ) _o −NH ₂ →PCD−NH−(CH ₂ ) _o −NH ₂ +RSO ₃ H As a diaminoalkane, the number of methylene groups is 2.
~20 types are used, but 26 types are preferred because they are liquid and easy to handle. If it is a solid, it is used after being dissolved in a solvent such as N,N-dimethylformamide or dimethyl sulfoxide. That is, a sulfonylated polycyclodextrin resin is dispersed in a diaminoalkane solution of 2 to 10 times its weight, or a solution diluted with a solvent, and the mixture is maintained at a temperature of 50° C. to 150° C. and allowed to react while being slowly stirred. To prevent oxidation of the diaminoalkanes, the reaction is carried out under a nitrogen stream if necessary.
The reaction is usually completed in 1 to 5 hours, and a polycyclodextrin resin having diaminoalkyl groups is obtained. The reaction product is separated, then thoroughly washed with a lower alcohol and then dried. The thus obtained polycyclodextrin resin having aminoalkylamino groups has a structure as shown in Figure 1, and since it has primary and secondary amino groups, it can adsorb anions and is used as an anion exchanger. Not only can it be used, but enzymes, peptides, etc. can be bound by treatment with diepoxy compounds, dialdehyde compounds, etc., so it can be used as immobilized enzymes and carriers for affinity chromatography.

Next, the present invention will be explained in more detail with reference to Reference Examples and Examples.

Reference example: 25g of powdered β-cyclodextrin and 12.5g of water
ml and knead, 30% aqueous sodium hydroxide solution
Add 37.5 ml and 100 mg of sodium borohydride to make a homogeneous solution, keep it at 50°C, add 7.5 ml of epichlorohydrin dropwise while stirring at 200 rpm, stir for 30 minutes, and then remove the bubbles.
Keep at 50 °C for 30 min. Add 375ml of liquid paraffin to the resulting syrupy liquid and keep it at 50℃.
Stir at 1000 rpm for 5 minutes to uniformly disperse, then add 25 ml of epichlorohydrin and stir for another 2 hours.
The mixture was kept at 50°C and stirred. The obtained beads were separated by filtration, washed three times with 100 ml of hexane, then three times with 100 ml of acetone, washed with distilled water until the pH of the washing solution became neutral, and then soaked in distilled water 1 for 1 day. water-soluble components were removed. After washing with 50% acetone water, it was immersed in 300 ml of acetone for 1 day to dehydrate it, dried at 70°C for 1 day, and further dried with phosphorus pentoxide in a vacuum desiccator for 1 day. By sieving 29 g of the poly-β-cyclodextrin beads obtained in this way,
26g of 200 mesh beads were obtained.

Example 1 10 g of beaded poly-β-cyclodextrin resin obtained in Reference Example and 10 g of p-toluenesulfonyl chloride were dispersed in 150 ml of dehydrated pyridine,
The mixture was left to react at room temperature for one week. The reaction product was separated, thoroughly washed with ethanol and then water until the pyridine odor disappeared, then dehydrated with ethanol and vacuum dried to obtain 14 g of bead-shaped polycyclodextrin resin having p-toluenesulfonyl groups. It was done.

2 g of this resin was dispersed in 10 ml of ethylenediamine and reacted for 1 hour while being kept at 100° C. in a nitrogen stream. The reaction product is ethanol, then distilled water,
The sample was washed until the pH of the water became neutral, dehydrated with ethanol, and then dried in vacuum. In this way, 1.3 g of polycyclodextrin resin having aminoethylamino groups was obtained. Take 1g of this and 1N
Titration with hydrochloric acid revealed that it contained 1.9 x 10 ^-3 moles of aminoethylamino groups.

Therefore, 1 g of this resin has the ability to adsorb 3.8×10 ⁻³ equivalents of anions and can be used as an anion exchanger.

Moreover, since it is an insoluble resin, it can be used repeatedly by washing.

Example 2 Bead-shaped polycyclodextrin resin 2 having p-toluenesulfonyl group obtained in Example 1
g was dispersed in 10 g of 1,5-diaminopentane, kept at 100° C. in a nitrogen stream, and reacted for 2 hours. The reaction product was washed with ethanol and then with distilled water until the pH of the water became neutral, dehydrated with ethanol, and then vacuum-dried. Thus, 1.4 g of polycyclodextrin resin with aminopentylamino groups
was gotten. When 1 g of this was taken and titrated with 1N hydrochloric acid, it was found to contain 1.9 x 10 ^-3 moles of aminopentylamino groups.

Therefore, 1 g of this resin can adsorb 3.8×10 ⁻³ equivalents of anions and can be used as an anion exchanger.

Moreover, since it is an insoluble resin, it can be used repeatedly by washing.

[Brief explanation of the drawing]

Deduced structures of polycyclodextrin resin and polycyclodextrin resin having aminoalkylamino groups. Polycyclodextrin resin has a crosslinking agent bonded to the hydroxyl group of cyclodextrin, and is presumed to have a network structure as a whole. The aminoalkylamino group is attached to the hydroxyl group of the cyclodextrin and the crosslinking chain. A: Estimated structure of polycyclodextrin resin,
B: Estimated structure of polycyclodextrin resin having aminoalkylamino group, C: Structure of cyclodextrin unit, n-6: α-cyclodextrin n-7: β-cyclodextrin n-
8: γ-cyclodextrin, X: aminoalkylamino group.

Claims (1)

[ _Scope of Claims] 1. After crosslinking cyclodextrin with epichlorohydrin or diepoxy compound and swelling the polycyclodextrin resin in which the reaction product is insolubilized in an organic solvent, R is an alkyl group or an aryl group) A sulfonyl chloride represented by the following formula is reacted to introduce a sulfonyl group, and then the general formula NH ₂ -(CH ₂ )n-NH ₂ (n in the formula is 2 to 20) is introduced. 1. A method for producing a polycyclodextrin-based resin having an aminoalkylamino group, which comprises substituting a sulfonyl group with an aminoalkylamino group by reacting a diaminoalkane represented by the following formula: 2. The manufacturing method according to claim 1, wherein the sulfonyl chloride is p-toluenesulfonyl chloride, and the diaminoalkane is ethylenediamine and 1,5-diaminopentane.