JP2000344835A - Stimulus-responsive polymeric material and its preparation - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a polymeric compound capable of showing temperature response in a solution and utilizable as a material having the function of adsorbing, separating, or releasing a substance, which compound comprises repeating units being units derived from a hydroxylalkylamide monomer having a large number of carbon atoms. SOLUTION: Provided is a polymeric compound comprising repeating units of the formula (wherein Z is H or methyl; X is H, a 1-8C linear or branched structure, or an aliphatic hydrocarbon group having a hydroxyl group in at least one position; Y is a 2-8C linear or branched aliphatic hydrocarbon group having a hydroxyl group in at least one position, provided that X and Y may be combined to form a chemical bond; and n is an integer of 2 or greater). The monomer from which each repeating unit is derived is exemplified by a compound synthesized from the reaction of acrylic chloride, methacrylic chloride, acrylic anhydride or methacrylic anhydride with a 3-16C linear, branched, or cyclic alkylaminoalcohol.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a polymer compound which causes the polymer itself to expand and contract and agglomerate together with a change in the polarity of the polymer itself due to a physical stimulus such as a temperature change, a process for producing the polymer, The present invention relates to a material having separation, adsorption and release functions and a method for separating a substance using the compound.

[0002]

2. Description of the Related Art Known as a temperature-responsive polymer,
N-alkylacrylamide represented by N-isopropylacrylamide is widely used for application to DDS and a separating agent. However, there is only an alkyl group having a small number of carbon atoms in the side chain of an alkylacrylamide-based monomer exhibiting temperature response. For this reason, it is difficult to efficiently separate, adsorb, and release all biological components, organic substances, and the like, because of their low hydrophobicity or low hydrogen bonding property. Further, by obtaining a copolymer using a monomer having a hydrogen-bonding group, a hydrogen-bonding group can be introduced, but in this case, the cloud point becomes high or the temperature responsiveness disappears, In particular, separation, adsorption and release of biological components are severe.

[0003]

SUMMARY OF THE INVENTION It is an object of the present invention to provide a polymer having a side chain having a large number of carbon atoms for imparting hydrophobicity and having a functional group having a hydrogen bonding property. Is to provide molecules.

[0004]

Means for Solving the Problems The present inventors have synthesized a hydroxyalkylamide monomer having a large number of carbon atoms in order to synthesize a thermoresponsive polymer having high hydrogen bonding and high hydrophobicity. However, it was found that a polymer material having a hydroxyl group having hydrogen bonding properties and a hydrophobic group by polymerizing the polymer exhibits temperature responsiveness. In addition, the inventor
By utilizing a material for adsorbing, separating and releasing a substance containing such a temperature-responsive polymer compound, the present inventors have found application to the separation of various substances. The present invention has been completed based on such findings.

That is, the present invention provides a compound of the formula I

[0006]

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(Wherein Z represents a hydrogen atom or a methyl group. X represents a hydrogen atom or an aliphatic hydrocarbon group having a linear or branched structure having 1 to 8 carbon atoms and having at least one hydroxyl group.) , Y represents an aliphatic hydrocarbon group having a linear or branched structure having 2 to 8 carbon atoms and having one or more hydroxyl groups, and in some cases, X and Y may form a chemical bond. Is an integer of 2 or more.) A polymer compound having a repeating unit represented by the following formula, or a copolymer and a gel structure containing this unit structure, and a polymer compound exhibiting temperature responsiveness in a solution; The present invention relates to a method and a material for separating, adsorbing and releasing a substance using the method.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION The polymer compound according to the present invention has the following structure.

[0009]

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(Wherein Z represents a hydrogen atom or a methyl group. X represents a hydrogen atom or an aliphatic hydrocarbon group having a linear or branched structure having 1 to 8 carbon atoms and having at least one hydroxyl group.) , Y represents an aliphatic hydrocarbon group having a linear or branched structure having 2 to 8 carbon atoms and having one or more hydroxyl groups, and in some cases, X and Y may form a chemical bond. Represents an integer of 2 or more.) Or a polymer material comprising a repeating unit alone or a copolymer and a gel structure containing this repeating unit structure and exhibiting a temperature responsiveness in an aqueous solution system. .

In the present invention, the aliphatic hydrocarbon group having at least one straight-chain or branched hydroxyl group having 1 to 8 carbon atoms refers to a straight-chain or branched hydroxylalkyl group having 1 to 8 carbon atoms. The aliphatic hydrocarbon group having one or more hydroxyl groups having a linear or branched structure having 2 to 8 carbon atoms refers to a linear or branched hydroxylalkyl group having 2 to 8 carbon atoms.

In the present invention, "X and Y form a chemical bond" means that X and Y each have an aliphatic hydrocarbon group, and the total number of carbon atoms of X and Y is in the range of 5 to 16, and the chemical bond forms a hydroxyl group. In one or two or more locations. The fact that X and Y are chemically bonded means that X and Y are covalently bonded.

The number of hydroxyl groups contained in the hydroxylalkyl group is one or two or more. Preferred hydroxyalkyl groups include 1-hydroxypentyl, 2-hydroxypentyl, 3-hydroxypentyl, tran
s-hydroxycyclohexyl group, 6-hydroxyhexyl group, 2-hydroxy-3-methylpentyl group, 5-
Hydroxy-3-ethylpentyl group, 3-hydroxyhexyl group, 7-hydroxyheptyl group, 6-hydroxyheptyl group, 8,3-dihydroxyoctyl group, 8.5
-Dihydroxyoctyl group and the like.

N is not particularly limited as long as it is an integer of 2 or more, but is preferably in a range where the molecular weight is 700,000 or less. A particularly preferred molecular weight range is from 1,000 to
700,000.

The monomers represented by the above formula (I) include:
Acrylic acid chloride, methacrylic acid chloride, acrylic acid anhydride, or a compound synthesized by the reaction of methacrylic anhydride with an alkylamino alcohol can be exemplified. The alkylamino alcohol in the present invention is an aliphatic hydrocarbon having a linear or branched structure having 3 to 16 carbon atoms, having one or more hydroxyl groups, or having 1 to 3 hydroxyl groups having a cyclic structure having 3 to 16 carbon atoms. It is an alicyclic hydrocarbon group having more than one site. Preferred alkylamino alcohols are those having one or more hydroxyl groups having 1 to 12 carbon atoms, such as 4-aminopentanol, 5-aminopentanol, 3-aminopentanol, 2-aminopentanol, and trans-amino. Pentanol, 6-aminohexanol, N-5-hydroxypentyl-N'-methyl-8-amino-3,5-dihydroxyoctyl and the like can be mentioned.

In the present invention, the copolymer containing the above repeating unit is defined as a monomer represented by the above chemical formula and another kind of t
-Butylacrylamide, n-butylacrylamide,
Alkyl acrylamide such as i-butyl acrylamide, acrylamide, hexyl acrylamide, heptyl acrylamide, or alkyl methacrylamide such as t-butyl methacrylamide, n-butyl methacrylamide, butyl methacrylamide, hexyl methacrylamide, heptyl methacrylamide, or Methacrylic acid, alkyl acrylates such as n-butyl acrylate, s-butyl acrylate, t-butyl acrylate, n-propyl acrylate, i-propyl acrylate, or methyl methacrylate, n-butyl methacrylate, s Alkyl methacrylates such as -butyl methacrylate, t-butyl methacrylate, n-propyl methacrylate, i-propyl methacrylate,
Or hydroxyethyl methacrylate, hydroxyethyl acrylamide, 2-aminoethyl methacrylamide, aminostyrene, 2- (t-butylamino) ethyl methacrylate, 2-sulfoethyl methacrylate, 3
-Sulfopropyl acrylate, hydroxy group such as glycidyl methacrylate, amino group, sulfone group, a monomer having a functional group such as an epoxy group, or a random copolymer with a monomer such as styrene, It means a block copolymer.

The gel structure in the present invention means a gel structure obtained by reacting with a crosslinking agent such as methylenebisacrylamide. The polymer compound of the present invention is produced as follows. One or two or more monomers used as raw materials of a polymer compound and a polymerization initiator are dissolved in a polymerization solvent, and a polymerization reaction is started by heating or the like. At this time, a bifunctional monomer may be dissolved in order to obtain a gel structure containing the polymer compound. Further, at this time, a chain transfer agent may be dissolved in the polymerization solvent in order to adjust the molecular weight of the polymer compound or to introduce a reactive functional group into a terminal of the polymer compound. After the polymerization reaction, the polymer is reprecipitated in a solvent in which the polymer is not dissolved, whereby a stimulus-responsive polymer compound at a desired temperature can be obtained.

The polymer compound of the present invention can be immobilized on the surface of a carrier such as silica gel or a polymer gel by utilizing a reactive functional group introduced into the terminal. Alternatively, after immobilizing a polymerization initiator on a solid surface such as silica gel or a polymer, one or two or more types of monomers used as a raw material of the polymer compound are dissolved in a polymerization solvent to immobilize the polymerization initiator. By initiating the polymerization reaction by heating or the like in the presence of a carrier such as silica gel or polymer gel, such a polymer compound can be immobilized on the surface of the carrier such as silica gel or polymer gel. At this time, a bifunctional crosslinking agent may be dissolved in order to obtain a gel structure of the polymer compound. Further, at this time, a chain transfer agent may be dissolved in the polymerization solvent to adjust the molecular weight of the polymer compound or to introduce a reactive functional group. The material containing such a high molecular compound can be applied to various liquid chromatography carriers, adsorption / separation carriers such as adsorbents, releasing agents such as bioproducts, and biofunctional materials.

[0019]

The present invention will be described in more detail with reference to the following experimental examples, but the present invention is not limited to these examples.

[0020]

Example 1 5 parts in dimethylformamide (140 ml)
-Aminopentanol (5.4 g) and triethylamine (5.6 ml) were added, and a mixed solution of dimethylformamide (30 ml) in which acrylic acid chloraloid (4.2 ml) was dissolved was added at -40 ° C. Then, the mixture was stirred while being dropped. After 2 hours, the solution was filtered to remove the precipitate, and then the solvent was removed by a rotary evaporator. After removing the solvent, the residue was dissolved in acetone, the eluent containing the target substance was separated by a silica column, and the solvent was removed with a vacuum pump to purify 5-hydroxypentylacrylamide (2.1 g).

The obtained 5-hydroxypentylacrylamide (1.0 g) and 4,4'-azobis (4-cyanovaleric acid) (20 mg) as a polymerization initiator were added to dimethylformamide (5 ml), and the mixture was freeze-thawed. After degassing, radical polymerization was performed at 65 ° C. for 3 hours.
After the polymerization reaction, dimethyl sulfoxide (7 m
l) was added to the reaction solution, and this mixed solution was reprecipitated in a mixed solution of acetone-ether = 1: 3 (volume ratio) to obtain a desired homopolymer of poly (5-hydroxypentylacrylamide). (0.51 g). The number average molecular weight (Mn) of the polymer obtained at this time was 2
000 and a molecular weight distribution (Mw / Mn) of 2.7.
It was 7.

0.1% by weight of this homopolymer in aqueous solution
And the change in transmittance due to temperature change was measured (FIG. 1). As a result, the cloud point is 4
It was shown to be around 0.8 ° C.

To immobilize the polymer on a chromatography support, dissolve the polymer (0.3 g), hydroxysuccinimide (0.8 g) and dicyclohexylcarbodiimide (0.8 g) in dimethylformamide (30 ml). And stirred overnight at room temperature.
After the stirring, the deposited precipitate is sufficiently removed, reprecipitated in a mixed solvent of acetone: diethyl ether = 1: 3 (volume ratio), and the desired polymer chain terminal is obtained by drying under vacuum to obtain hydroxysuccinimide. A polymer was obtained. Next, the activated polymer (0.15 g) and aminopropyl silica (0.75 g) were added to dimethylformamide (30 ml) to immobilize the polymer on silica. After the immobilization, the same operation was performed under the same conditions to immobilize the polymer. From the result of the elemental analysis, it was confirmed that the polymer was immobilized because the organic matter content increased by 10 wt%.

This silica was packed in a 4.6 × 30 mm stainless steel column tube to separate the peptide.
It was confirmed that the retention time of peptides such as beta-endorphin, angioten and the like varies depending on the temperature in the aqueous mobile phase.

[0025]

Example 2 trans-Aminocyclohexanol (5.0 g) and triethylamine (6.1 ml) were added to chloroform (100 ml). In this mixed solution, acrylic acid chloraloid (3.4 ml) was added to chloroform (30 ml). The dissolved mixed solution was dropped over 2 hours in ice cooling to cause a reaction. After the reaction, after the dropwise addition, the mixture was stirred at room temperature for 1 hour, and then the solvent was removed with a rotary evaporator. After removing the solvent, ethyl acetate (250 m
l) was added, the precipitate was filtered, the filtrate was applied to a silica column to obtain an eluent containing the target substance, concentrated, concentrated, and recrystallized by adding n-hexane (250 ml). After drying, the desired 4-hydroxycyclohexylacrylamide (2.4 g) was obtained.

This monomer (0.8 g) was treated with 4,4′-azobis (4-cyanovaleric acid) (5 mg) as a polymerization initiator.
Together with dimethylformamide (5 ml)
After deaeration by the freeze-thaw method, at 65 ° C. for 3 hours,
Polymerization was performed. After the polymerization, dimethyl sulfoxide (7 ml) was added to this solution, and acetone: diethyl ether =
Reprecipitation was performed in a mixed solution of 1: 3 (volume ratio), and the precipitate was dried in vacuum to obtain the target poly (trans-hydroxycyclohexylacrylamide) (0.90 g). As a result of the molecular weight analysis by GPC, the number average molecular weight (Mn) of this homopolymer was 28,000 and the molecular weight distribution (Mw / Mn) was 2.68. The measurement of the transmittance according to the temperature change showed that the cloud point was around 41.2 ° C. The measurement of the number average molecular weight by the terminal titration method showed almost the same value.

[0027]

Example 3 5-hydroxypentylacrylamide (1.0 g) and t-butylacrylamide (0.12 g)
g) and 2,2′-azobisisobutylnitrile (12 m
g) was dissolved in dimethyl sulfoxide (6 ml) and
Polymerization was performed at 3 ° C. for 3 hours. After the polymerization reaction, acetone:
The desired copolymer was obtained by mixing in a mixed solution of ether (1: 3 (volume ratio)). 1 w of this copolymer polymer
It was dissolved in an aqueous solution so that the concentration became t%, and it was observed whether or not it showed a cloud point due to a change in temperature. Was done.

[0028]

Example 4 6-aminohexanol (2.0 g) and triethylamine (2.1 ml) were added to a chloroform solvent (8
0 ml), and into this solution, chloroform (2 ml) in which acrylic acid chloraroid (1.4 ml) was dissolved.
0 ml) The solvent was added dropwise over 3 hours under ice cooling. After the dropwise addition, the mixture was stirred at room temperature for 3 hours, and the solvent was removed with an evaporator to obtain a precipitate. Ethyl acetate (120 ml) was added to the precipitate, the precipitate was removed by filtration, and the filtrate was concentrated with an evaporator. After concentration, the concentrate was applied to a silica column to separate an eluate containing the target substance, which was concentrated and recrystallized to obtain the desired 6-hydroxyhexylacrylamide (1.4 g). Obtained, 6
-Hydroxyhexylacrylamide (1.0 g) and 2,2′-azobisisobutylnitrile (10 mg) were dissolved in dimethylformamide (5 ml), deaerated by freeze-thawing method, and then polymerized at 70 ° C. for 2 hours. The reaction was performed. After the polymerization reaction, the polymer was reprecipitated with a mixed solvent of tetrahydrofuran: ether = 1: 1. After reprecipitation, drying was performed under vacuum to obtain the target poly (6-hydroxyhexylacrylamide) (0.84 g). GPC analysis showed that the polymer had a number average molecular weight (Mn) of 9,600 and a molecular weight distribution (Mw / M
n) was 4.05.

This polymer was dissolved in an aqueous solution so as to have a concentration of 1 wt%, and a change in transmittance due to a change in temperature was measured (FIG. 3). In addition, in order to examine the temperature response in a guanidine aqueous solution, a polymer was dissolved at a similar concentration in a 1.0 M guanidine-hydrochloric acid aqueous solution, and a change in transmittance due to a temperature change was measured. As a result, 1.0 M guanidine-
It was found that the aqueous hydrochloric acid solution exhibited a cloud point at 5.2 ° C.

[0030]

Example 5 5-Hydroxypentylacrylamide (0.8 g), acrylamide (0.2 g) and 2,2 ′
-Azobisisobutylnitrile (10 mg) was dissolved in dimethylsulfoxide (7 ml), and polymerization was carried out at 70 ° C for 2 hours. After the polymerization reaction, the mixture was put into a mixed solution of acetone and ether (1: 3 (volume ratio)) to obtain a desired copolymer. The copolymer polymer was dissolved in an aqueous solution so as to have a concentration of 1 wt%, and it was observed whether or not a cloud point was exhibited by a change in temperature.
Turbidity at a temperature of, it was confirmed to have a cloud point.

[0031]

According to the present invention, it is possible to synthesize a thermoresponsive polymer having a large amount of hydroxyl groups and a high hydrophobicity as compared with a conventional amide-based thermoresponsive polymer. Noh can be controlled. In addition, since the polarity and hydrogen bonding properties also change depending on the molecular weight, concentration, density, etc. of the polymer, it exhibits adsorption, separation, and release behavior for substances such as bioproducts that could not be obtained with conventional thermoresponsive polymers. It is possible.

[Brief description of the drawings]

FIG. 1 is a diagram in which poly (5-hydroxypentylacrylamide) has exhibited temperature responsiveness in an aqueous solution.

FIG. 2 is a diagram in which poly (trans-hydroxycyclohexylacrylamide) has exhibited temperature responsiveness in an aqueous solution.

FIG. 3 is a diagram in which poly (6-hydroxyhexylacrylamide) has exhibited temperature responsiveness in an aqueous solution.

 ────────────────────────────────────────────────── ─── Continued on the front page (72) Inventor Mitsuo Okano 6-12-12 Kokudaidai, Ichikawa-shi, Chiba F-term (reference) 4D017 AA03 BA03 CA13 CB01 DA02 DA03 EA01 4J100 AB02Q AJ02Q AL03Q AL09Q AM15Q AM17Q AM21P AM21Q BA03P BA03Q BA16P BA29P BA29Q BA40P BA56Q BC54Q CA01 CA04 DA24 DA61 EA03 FA03 JA15 JA17 JA51

Claims (16)

[Claims] 1. A compound of the formula I (In the formula, Z represents a hydrogen atom or a methyl group. X represents a hydrogen atom or an aliphatic hydrocarbon group having a linear or branched structure having 1 to 8 carbon atoms and having at least one hydroxyl group, and Y is Having a linear or branched structure having 2 to 8 carbon atoms,
It represents an aliphatic hydrocarbon group having one or more hydroxyl groups. In some cases, X and Y may form a chemical bond.
n shows the integer of 2 or more. A) a polymer compound containing a repeating unit represented by the formula (1) and showing temperature responsiveness in a solution. 2. The compound according to claim 1, wherein the monomer represented by the formula I is a compound synthesized by reacting acrylic acid chloride, methacrylic acid chloride, acrylic anhydride or methacrylic anhydride with an alkylamino alcohol. High molecular compounds. 3. The polymer compound according to claim 1, which comprises only a repeating unit represented by the formula (I). 4. The polymer compound according to claim 1, which contains a repeating unit represented by the formula I and has a copolymer structure. 5. The polymer compound according to claim 1, wherein a terminal of the polymer material has a functional group such as a carboxyl group, an amino group, a hydrogen group, or a cyano group. . 6. The cloud point of the polymer material is determined by the salt concentration in the aqueous solution,
The concentration of the polymer, the pH concentration, the concentration of the organic solvent, the size of the molecular weight of the polymer, and the number of the cationic group or the anionic group in the copolymer are controlled by one or two or more. Item 6. The polymer compound according to any one of Items 1, 2, 3, 4, and 5. 7. The polymer compound according to any one of claims 1, 2, 3, 4, 5, and 6, wherein the polymer material is in the form of a polymer gel. 8. The material according to claim 1, wherein the temperature-responsive material changes its hydration state via polarity and hydrogen bonding at the cloud point. The polymer compound according to claim 1. 9. A polymer capable of converting a hydroxyl group of the polymer compound according to claim 1 into another type of functional group such as an amino group or a carboxyl group by a chemical reaction, and exhibiting temperature responsiveness in a solution even after the conversion. Compound. 10. The method of claim 1, 2, 3, 4, 5, 6, 7,
The polymer material according to any one of 8 and 9 is immobilized on a filler, and a polar or hydrogen bond interacting with a target substance can be produced at a constant temperature by using the filler. A method for separating a substance, comprising separating, adsorbing, and releasing a substance by utilizing the fact that the polarity or hydrogen bonding property of the material is changed. 11. The separation method according to claim 10, wherein the substance is a biological component or an organic compound. 12. The method according to claim 11, wherein the biological component is any one of a protein, a nucleic acid, a peptide, and a cell. 13. The method according to claim 11, wherein the organic compound is a medicament. 14. The method of claim 1, 2, 3, 4, 5, 6, 7,
10. The polymer material exhibiting temperature responsiveness according to any one of 8 and 9, which is immobilized on a filler, and using the filler, an organism such as a protein, a nucleic acid, a peptide, or a cell characterized by a temperature change is used. Separation, adsorption, and release materials that separate, adsorb, and release components or organic compounds. 15. The method of claim 1, 2, 3, 4, 5, 6, 7,
10. A packing material for chromatography using the polymer material exhibiting temperature response according to any one of 8 and 9. 16. A monomer having a repeating unit according to claim 1 or a monomer and another monomer dissolved in a solution together with a polymerization initiator, and a polymerization reaction is carried out by heat or light. 2. The method for producing a polymer compound according to claim 1, wherein the method is performed.