JPS5850621B2 - Polyion complexes - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides a polyionic complex (hereinafter referred to as PIC) produced from a strong electrolyte type polyanionic polysaccharide and a polycationic polymer containing a nucleobase or its related compound.

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In the production of resins for the separation and purification of biochemical-related compounds, such as nucleic acid-related substances, polysaccharides, especially cellulose, can be obtained at low cost and at the same time have a similar biological composition (JJ properties), so various derivatives can be manufactured. has been done.

Furthermore, various polymers containing both a nucleic acid base and a polysaccharide have been produced for the purpose of separating and purifying nucleic acid-related substances.

The method involves polymerizing vinyl monomers of tacleoside.

Therefore, only low molecular weight products can be obtained.

Also, the production of monomers is quite complex and there are still many problems.

In order to solve these problems, the inventors of the present invention have conducted extensive research, and as a result, we have developed a polyion complex of a strong electrolyte type polyanionic polysaccharide and a polycationic polymer containing a nucleobase or its related compound. I found out what to do.

The present invention is based on the following general formula (where R1 and R4 are a hydrogen atom or an alkyl group having 1 to 4 carbon atoms).

R2 and R3 are alkyl groups having 1 to 4 carbon atoms.

B is a halogen atom or a hydroxyl group.

y is the number O or 1. X is any number including O.

a is an arbitrary number of 10 or more.

b is a number of 1 or 2. n is a number of 10 or more.

X is adenine, thymine, cytosine, uracil or ethoxypyrimidine.

The purine ring reacts at the 91st position, and the piIJ midine ring reacts at the 11th position.

M is a vinyl monomer that does not become a polymer electrolyte.

) A polycationic polymer containing at least one kind of adenine, thymine, cytosine, uracil or ethoxypyrimidine selected from the group shown in ) and sodium alginate, heparin, an alkali metal salt of carboxymethylcellulose, sulfated cellulose or sulfated methylcellulose The present invention relates to a method for producing a polyion complex, characterized in that at least one strong electrolyte type polyanionic polysaccharide selected from the following is reacted in an aqueous solution or an organic solvent miscible with water.

According to the method of the present invention, an insoluble polymer containing simultaneously a polysaccharide and a nucleobase or a related compound thereof can be produced very easily.

The strong electrolyte type polyanionic polysaccharide used in the present invention means a polymer electrolyte having a strong electrolyte type anion group in the polysaccharide molecule, and these include natural ones such as sodium alginate, Heparin, etc.; examples of semi-synthetic acids include sodium salt of carboxymethylcellulose, sulfated cellulose, sulfated methylcellulose, etc.

On the other hand, the polycationic polymer used in the present invention must contain a nucleobase or a compound related thereto, and is specifically represented by the following general formula.

(However, R1 and R4 are a hydrogen atom or an alkyl group having 1 to 4 carbon atoms.

R2 and R3 are hydrocarbon groups having 1 to 4 carbon atoms.

B is...a rogen atom or a hydroxyl group.

y is the number O or 1. X is any number including O.

a is an arbitrary number of 10 or more.

b is a number of 1 or 2. n is a number of 10 or more.

X is adenine, thymine, cytosine, uracil or ethoxypyrimidine.

The purine ring reacts at the 91st position, and the pyrimidine ring reacts at the 11th position.

M is a vinyl monomer that does not become a polymer electrolyte.

) M in general formulas (1) to 3) is a vinyl monomer that does not become a polymer electrolyte, and is an aromatic vinyl monomer,
Substituted styrenes such as styrene, α-methylstyrene, nitrogen-containing heterocyclic monomers such as vinylpyridine: Aliphatic vinyl monomers, such as acrylonitrile, substituted acrylonitriles such as mequacrylonitrile, vinyl monomers in the broad sense of acrylic acid or nuclear esters I can give my body away.

PIC obtained by the present invention is a new substance, and can be produced by the reaction of the above-mentioned strong electrolyte type polyanionic polysaccharide with the above-mentioned nucleobase or related compound-containing polycationic polymer. It is something that you have.

The PIC according to the present invention can be produced by dissolving a polyanionic polymer and a polycationic polymer in appropriate solvents, and then reacting the solutions.

In this case, the solvent that can be used is water or an organic solvent that is miscible with water, and examples of the organic solvent include alcohol,
For example methanol, ethanol; ketones such as acetone; cyclic ethers such as tetrahydrofuran:
Examples include nitrogen-containing or sulfur polar solvents, such as N-N-dimethylformamide and N-N-dimethylacetamide.

The method for producing PIC is simple; it is sufficient to dissolve the polyanionic polymer and the polycationic polymer in the above-mentioned solvents, and then mix the two solutions.

The purpose of the present invention can also be achieved by adding a solid substance directly into one of the solutions, but this may result in non-uniform composition of the resulting polyion complex, poor reproducibility, and problems in the purification stage*. It takes time.

The concentration of polycationic and polyanionic polymers used in this invention can vary from 0.01 to 70% by weight.

Preferably it is 0.1 to 15% by weight.

The temperature at which the two solutions are reacted can range from 0°C to 90°C, but preferably from room temperature to 50°C.

The equivalent ratio of polyanion to polycation ranges from 0.05 to 20.

Due to its structure, the polyion complex produced by the present invention is an extremely effective material for resins for separating and purifying nucleic acid bases, nucleosides, nucleotides, or their related compounds, and can also be used as a material for polyion complex membranes. It has many uses.

The present invention will be explained in more detail below using Examples, but is not limited to these Examples.

Example 1 A polycationic polymer represented by the following formula (poly-4-vinylpyridine (molecular weight 1X10") and 1-(2'hydroxy-
37-Bromopropyl)-thymintho was obtained by heating in N-N'-dimethylformamide.

) and dissolved it in 450wt1 of distilled water.

On the other hand, a solution was prepared by dissolving sodium alginate (1000 cps/1% aqueous solution) 41 in 500 rul of distilled water.

This solution was slowly added to the polycationic polymer solution at room temperature with vigorous stirring.

3, a white precipitate was formed at the same time.

After standing for a day and night, the precipitate was collected by filtration, washed with a large amount of water, then washed with acetone, and dried under vacuum, resulting in a yield of 8 tons of PIC.

The infrared absorption spectrum of the PIC obtained has 1650c.
Absorption based on thymine and ether bonds were observed at m and 1200 to 1050 crrL-1, respectively.

Example 2 A polycationic polymer represented by the following formula (poly-4-vinylpyridine (molecular weight 1 x 105) and 1-(2'hydroxy-3'-bromopropyl)-uracil) was placed in a 21 pressure-resistant beaker. '-obtained by heating in dimethylformamide.

)7. FW was added and dissolved in 800rrLl of distilled water.

On the other hand, sodium carboxymethyl cellulose (100
-300 cps)/1% aqueous solution) A solution was prepared by dissolving 4.51 in 500 ml of distilled water.

By reacting and treating in the same manner as in Example 1, PIC9
, got 5P.

The infrared absorption spectrum of the obtained polyion complex contains uracil at 1640 CrrL' and uracil at 1200~
Absorption based on ether bonds was observed at 1050cIrL-1.

Example 3 A polycationic polymer represented by the following formula (holJ-hinylbenzyldimethylamine-styrene copolymer (molecular weight 7X105) and 1-(2'-hydroxy-3'-7" rompropyl)--y-mine and N・N
Obtained by heating in '-dimethylformamide.

)16′? was dissolved in distilled water 11. On the other hand, 71 sodium alginate (same type as in Example 1) was added at 800 r.
fLl was dissolved in distilled water.

While stirring vigorously at room temperature, an aqueous solution of a polyanionic polymer was slowly dropped into the prepared aqueous solution of a polythionic polymer.

A white precipitate was formed simultaneously with the dropping.

After standing for a day and night, the precipitate was filtered and collected.

Wash the precipitate with a large amount of water, then with acetone,
As a result of drying under vacuum, PIC18.5f was obtained.

Examples 4 to 6 Reactions and treatments were carried out in the same manner as in Example 1, except that the polycationic and polyanionic polymers shown in Table I were used.

The reaction conditions and reaction results are shown in Table I. , Example 4.
The polycationic polymer of 5.6 is polyvinylbenzyldimethylamine (molecular weight 44 x 104) and 9-(2
'-chloroethyl)-adenine, poly-dimethylaminomethyl acrylate (molecular weight 1.2X104)
and 1-(2'-hydroxy-3'-bromopropyl)
- Thymine, poly-4-vinylpyridine (molecular weight 1x10
”) and 1-(2′-hydroxy-3′-7” romflovir)-4-ethoxypyrimidine, each with N
-N'-dimethylformamide.

Claims (1)

[Scope of Claims] 1 The following general formula (where R1 and R4 are a hydrogen atom or an alkyl group having 1 to 4 carbon atoms. R2 and R3 are an alkyl group having 1 to 4 carbon atoms. B is a halogen atom or a hydroxyl group. y is O or a number of 1. X is an arbitrary number including O. a is an arbitrary number of 10 or more. b is a number of 1 or 2. n is 10 The above numbers are as follows. A polycationic polymer containing at least one kind of adenine, thymine, cytosine, uracil or ethoxypyrimidine selected from the group shown in ), sodium alginate, heparin, an alkali metal salt of carboxymethylcellulose, sulfuric acid. 1. A method for producing a polyion complex, which comprises reacting at least one strong electrolyte type polyanionic polysaccharide selected from cellulose cellulose or methyl cellulose sulfate in an aqueous solution or an organic solvent miscible with water.