JPH09302034A - Polymeric material which can remove nucleic acid derivative and urea by adsorption - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a polymeric material which can effectively remove nucleic acid derivatives and urea from a hydrophilic solvent by forming specified residues. SOLUTION: Nucleic acid derivatives representing DNA or RNA constituents including nucleotides, nucleosides and nucleic acid bases, uric acid, xanthine and theobromine are entrapped in a polymer by forming complementary hydrogen bonding as shown in Fig in a hydrophilic solvent. The hydrophilic solvent herein mentioned means water or an organic solvent miscible with water in a volume ratio of 1:1, may be a mixture of at least two solvents in an arbitrary mixing ratio and is desirably a methanol/ethanol mixed solvent. It is desirable that the polymeric material which can remove nucleic acid derivatives and urea in a hydrophilic solvent has a main chain into the side chains of which residues represented by the formula (wherein X1 and X2 are each N or CH; Y is H or NHR2 ; R1 and R2 are each H, an alkyl, a phenyl or an acyl) are introduced through covalent bonds, and the main chain is a polyethylene, a polypeptide, a polyester or a polyether.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a polymeric material capable of efficiently adsorbing and removing a nucleic acid derivative and urea from a hydrophilic solvent system.

[0002]

Nucleic acid derivatives and urea are present in the blood as one of the final products of the metabolic system, and are eventually excreted as urine from the body. When present in blood, it causes various diseases. For example, uric acid, a type of nucleic acid derivative, causes hyperuricemia and gout. Thus, the development of a technique for selectively removing nucleic acid derivatives and urea is medically important, and it is particularly necessary to remove nucleic acid derivatives such as urea and uric acid from the body in dialysis treatment of patients with renal impairment. was there. The hollow fibers used for conventional dialysis treatment are mainly cellulosic fibers with fine pores, and substances unnecessary in the blood can be filtered by sieving the molecular size through the above pores. It was done. That is, since the material itself does not have the ability to recognize nucleic acid derivatives, the separation efficiency is poor, and a dialysis treatment requires a large-scale dialysis machine and a long time.

On the other hand, many attempts have been made to selectively bind and recognize biologically relevant substances such as these nucleic acid derivatives by hydrogen bonds. Rebek et al., "Journal, Of, American, Chemical, Society" (Journ
al of American Chemical S
The compounds described in Journal of the Society, Vol. 113, p. 5125, 1991 were synthesized as part of an attempt to bind and recognize a nucleic acid derivative by the above hydrogen bond, but most of them are nonpolar solvents such as chloroform. However, it did not efficiently remove the nucleic acid derivative and urea by hydrogen bonding in an aqueous solution. Hamilton et al., "Journal of American, Chemical, Society" (Journal of American)
Chemical Society, 1st 1993.
It is described that the compound described in Vol. 15, page 369 is capable of recognizing organic molecules even in a mixed solution of an organic solvent and water, but in a solvent system having a low ratio of the organic solvent, the recognition ability is almost lost. . Therefore, it has been desired to develop a polymer material capable of efficiently binding and recognizing a nucleic acid derivative in a hydrophilic solvent.

[0004]

An object of the present invention is to obtain a polymeric material capable of efficiently removing a nucleic acid derivative and urea in a hydrophilic solvent.

[0005]

[Means for Solving the Problems] The above object is to provide a polymeric material capable of adsorbing and removing a nucleic acid derivative and urea in a hydrophilic solvent, wherein the polymeric material has a residue represented by the formula (1). Was achieved by a polymeric material characterized by: Here, X ₁ , X ₂ = N, CH Y = H, NHR ₂ R ₁ , R ₂ = H, an alkyl group, a phenyl group, and an acyl group are shown.

The term "nucleic acid derivative" as used herein refers to nucleotides, nucleosides and nucleobases constituting DNA or RNA, and uric acid, xanthine and theobromine. These nucleic acid derivatives are incorporated into a polymer by forming complementary hydrogen bonds as shown in FIG. 1 in a hydrophilic solvent. Therefore, the functional group in Formula 1 is
It may be appropriately selected so that the nucleic acid derivative to be removed is complementary in hydrogen bonding mode. For example, uracil, uridine,
When there are three complementary hydrogen bond pairs such as thymine, thymidine, uric acid, xanthine and theobromine, Y = N
HR ₂ is preferred. R ₁ and R ₂ may be any functional groups, since one hydrogen atom may be present in each bond for forming a hydrogen bond pair, but preferably H, an alkyl group, a phenyl group, an acyl group, It is more preferably H. On the other hand, like adenine or adenosine, 2
When it has two hydrogen-bonding capacities, Y = H is preferable.

FIG. 1

The hydrophilic solvent of the present invention means water or an organic solvent which can be mixed with water in a volume ratio of 1: 1, and examples thereof include methanol, ethanol, propanol, butanol, acetonitrile, tetrahydrofuran and dimethyl sulfoxide. , Dimethylformamide and the like. The mixing ratio of these solvents may be arbitrary, and two or more kinds of solvents may be mixed. Of the above-mentioned solvents, water, methanol, ethanol or a mixed solvent thereof is preferable.

The present invention will be described in detail below. In the polymer material of the present invention, it is desirable that the residue represented by Formula 1 be introduced into the side chain of the polymer main chain by a covalent bond. The main chain may be polyethylene, polypeptide, polyester, or polyether. Further, the residue of Formula 1 may be directly linked to the main chain, or a spacer may be present between the main chain and the residue. The _{spacer, - (CH 2) n -} ,
_{- (C 6 H 4) -} , - CONH -, - COO -, - CO
_{O- (CH 2) n -,} - (C 6 H 4) - (CH 2)
_{n -, - COO- (CH 2} ) n -, - CONH- (CH
₂ ) _n −, and the like, but are not limited thereto. The aromatic ring of Formula 1 is a pyridine ring (X ₁ = X ₂
= CH), a pyrimidine ring (X ₁ = CH, X ₂ = N, or X ₁ = N, X ₂ = CH), a triazine ring (X ₁ = X ₂₎
= N) Either may be used. It can be appropriately selected depending on the synthesis method. If the residue of Formula 1 is contained in the polymer, another residue may be introduced into the same polymer. That is, a monomer containing Formula 1 and another monomer may be copolymerized. As the monomer used for the copolymerization, styrene, acrylamide, aminostyrene, vinyl acetate, vinyl benzoic acid, methyl acrylate, methyl methacrylate, cyanostyrene, acrylic acid, methylstyrene, tryptamide acrylate, naphthylethyl acrylate, acrylonitrile, And so on.

The preferred usage forms of the present invention are shown below. (1) A polymer material capable of adsorbing and removing a nucleic acid derivative and urea in a hydrophilic solvent, wherein the polymer material has a residue as shown in Formula 1. Here, X ₁ , X ₂ = N, CH Y = H, NHR ₂ R ₁ , R ₂ = H, an alkyl group, a phenyl group, and an acyl group are shown. (2) The polymer material according to (1), wherein R ₁ = R ₂ = H in the formula ₁ . (3) The nucleic acid derivative is uracil, uridine, thymine,
In the case of thymidine, uric acid, xanthine or theobromine, Y is NH ₂ , and the polymer material according to (2). (4) The polymer material according to (1) to (3), wherein the hydrophilic solvent is water, methanol and ethanol (5) The hydrophilic solvent is water, or a mixed solvent of water and methanol Or a mixed solvent of water and ethanol, wherein the polymer material according to (4) has a pH of 4 in the water or the mixed solvent in (5).
The polymer material according to (5), characterized in that it is used between 1 and 9.

The introduction of the residue of formula 1 into the polymer main chain may be carried out by a polymer reaction, or the corresponding monomer may be synthesized and polymerized. Further, it can be copolymerized with various monomers depending on the intended function. For example, Noguchi et al., "Organic Synthetic Chemistry", 1996, Vol.
By the method described on page 5, a diaminotriazine can be introduced by introducing a —CN group and then reacting with guanidine under basic conditions. Also, "Journal,
Of, American, Chemical, Society "(Jour
nal of American Chemical
The vinyl monomer containing a triazine ring can be previously synthesized by the method described in Sociity), Vol. 79, page 941, 1957, and this can be polymerized by a method such as radical polymerization or ionic polymerization. The obtained polymeric material can be used at any pH, but it is preferably 2 to 10, more preferably 4 to 9.

Specific examples of the residue introduced into the polymer are those shown in FIG.

FIG. 2

[0014]

The present invention will be described in more detail with reference to specific examples, but the present invention is not limited to the examples unless it exceeds the gist of the invention. Example 1 Synthesis of Polymer of the Present Invention A polymerization tube having a capacity of 50 ml was charged with commercially available 2-vinyl-4,6.
-Adding 2 g of diamino-1,3,5-triazine and 32 mg (?) Of azobisisobutyronitrile as a polymerization initiator, and further adding 50 ml of dimethyl sulfoxide as a solvent to dissolve the above-mentioned reagent, followed by freezing. Degas 3
The polymerization tube was vacuum-sealed repeatedly. This was polymerized by stirring in a hot water bath at 70 ° C. for 5 hours. The polymerization product was insoluble in the solvent dimethylsulfoxide, and thus was obtained as a precipitate as the polymerization proceeded. This is separated by filtration, further washed with a large amount of a solvent in the order of dimethylsulfoxide, water and methanol, and then dried under reduced pressure to obtain the polymer A (poly2-vinyl-4,6-diamino-1,
3,5-triazine) was obtained. The polymer A has a pH of 4
It was insoluble in substantial water between -8.

A polymerization tube having a volume of 50 ml was charged with 11 ml of styrene purified by vacuum distillation and 4 ml of acrylonitrile, and 15 ml of dimethylformamide was added as a solvent.
Furthermore, after adding 50 mg of azobisisobutyronitrile as a polymerization initiator, freeze deaeration was repeated 3 times to seal the polymerization tube in a vacuum. This was polymerized by stirring in a water bath at 70 ° C. for 5 hours. Immediately thereafter, the temperature was lowered to stop the polymerization, and the white precipitate obtained by pouring the above mixed solution into a large amount of methanol was separated by filtration and vacuum dried. 5 g of this copolymer was again added to dimethylformamide 3
The solution was dissolved in 0 ml, 1.7 g of dicyandiamide was added, 0.11 g of potassium hydroxide was further added as a catalyst, and the mixture was stirred at 130 ° C. for 1 hour to introduce a diaminotriazine ring. After the temperature was lowered to stop the reaction, the mixture was poured into a large amount of methanol again to obtain a yellowish white precipitate, which was filtered and vacuum dried to obtain a polymer B of the present invention. The amount of diaminotriazine introduced into the obtained polymer is
It was found from ¹ H-NMR to be 0.37 mmol / g.

Separation Experiment of Nucleic Acid Derivatives 1 mg of the obtained polymer was added to 1 ml of an aqueous solution (pH 5 to 8) of various nucleic acid derivatives having a concentration of 1.0 × 10 ⁻⁴ mol / liter, and 1 The temperature was kept at 20 ° C. After that, the polymer was removed by centrifugation, and the concentration of the remaining nucleic acid derivative in the supernatant was analyzed and quantified by the reversed phase high performance liquid chromatography method or the visible / ultraviolet spectrophotometer. Urea separation experiment 1 ml of urea aqueous solution with a concentration of 0.025 mol / liter
Then, 10 mg of the obtained polymer was added, and the mixture was kept at 20 ° C. for 1 hour with occasional shaking. After this, the polymer was removed by centrifugation and the supernatant was collected. Color was developed by adding an aqueous solution of p-dimethylbenzaldehyde hydrochloride thereto, and the concentration of urea was quantified from the absorbance at 430 nm.

[0017]

As shown in the table, the polymers A and B of the present invention showed a clear adsorption activity for the nucleic acid derivative and urea. Further, surprisingly, the polymer A of the present invention showed very high activity against uric acid, and all the uric acid in the aqueous solution was adsorbed on the polymer A. On the other hand, cellulose used as a raw material for hollow fibers did not show significant adsorption activity for nucleic acid derivatives.

Example 2 An experiment for separating nucleic acid and urea was conducted in the same manner as in Example 1 except that methanol was used as a solvent instead of water. As a result, polymers A and B
Efficiently removed urea and nucleobase derivatives even in methanol.

[Example 3] A separation experiment for nucleic acid and urea was carried out in the same manner as in Example 1 except that ethanol was used as a solvent instead of water. As a result, polymers A and B
Efficiently removed urea and nucleobase derivatives even in methanol.

As described above, the nucleic acid derivative and urea were efficiently removed from the hydrophilic solvent by using the polymer material of the present invention.

[Brief description of drawings]

FIG. 1 is a schematic diagram of a binding mode between a residue introduced into a polymer material of the present invention and a nucleic acid derivative or urea.

FIG. 2 is an example of residues that can be introduced in the present invention.

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[Procedure amendment]

[Submission date] September 30, 1996

[Procedure amendment 2]

[Document name to be amended] Drawing

[Correction target item name] All figures

[Correction method] Change

[Correction contents]

FIG.

[Fig. 2]

Continuation of front page (51) Int.Cl. ⁶ Identification code Office reference number FI Technical display area C08G 69/48 NRH C08G 69/48 NRH

Claims (3)

[Claims] 1. A polymeric material capable of adsorbing and removing a nucleic acid derivative and urea in a hydrophilic solvent, wherein the polymeric material has a residue as shown in formula (1). Here, X ₁ , X ₂ = N, CH Y = H, NHR ₂ R ₁ , R ₂ = H, an alkyl group, a phenyl group, and an acyl group are shown. 2. The polymer material according to claim ₁ , wherein R 1 in the formula 1 is H. 3. The polymer material according to claim 1 or 2, wherein the hydrophilic solvent is ethanol, methanol, or water.