JP2010255001A - Acrylamide derivative and polymer containing the same derivative - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a monomer and a polymer, or hydrogel for preparing the polymer or the hydrogel incorporating many functional groups while maintaining the sensitive temperature response. <P>SOLUTION: An acrylamide derivative represented by general formula (I) is used to prepare the polymer or copolymer, wherein X represents an amino group, a hydroxyl group, a halogen atom, a carboxyl group or -COOR<SP>4</SP>. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to an acrylamide derivative having a functional group in a side chain, a homopolymer and a copolymer of the derivative, and a crosslinked product of the polymer. The acrylamide derivative of the present invention can be easily radically polymerized to easily obtain a homopolymer, and can be copolymerized with other monomers. The copolymer synthesized from the acrylamide derivative of the present invention can be a temperature-responsive polymer that dissolves in the aqueous solution at the low temperature side and insolubilizes on the high temperature side at a specific temperature. Therefore, by immobilizing on the solid surface, it becomes hydrophilic on the low temperature side and hydrophobic on the high temperature side with a specific temperature as a boundary. When the copolymer of the present invention is used, not only changes in hydrophilicity and hydrophobicity are caused in a narrow temperature range, but also switching based only on temperature change from a charged state to an uncharged state becomes possible. Furthermore, since it has various functional groups, it is possible to introduce various molecules by using covalent bonds or hydrogen bonds.

  N-alkyl-substituted acrylamide polymer is one of temperature-responsive polymers. Among them, poly (N-isopropylacrylamide) (hereinafter abbreviated as PIPAAm) is below the phase transition temperature of 32 ° C. It shows water solubility, and above that, it rapidly insolubilizes and precipitates. A PIPAAm chain is introduced on the surface of a solid, the hydrophilicity / hydrophobicity is changed only by temperature change, and the cells are collected after cell culture or developed into chromatography. Such techniques are described in the following documents, for example.

T. Okano, N. Yamada, H. Sakai, Y. Sakurai, Journal of Biomedical Materials Research, 27, 1243-1251 (1993) T.Okano, N.Yamada, M.Okuhara, H.Sakai, Y.Sakurai, Biomaterials, 16, 297-303 (1995) H. Kanazawa, K. Yamamoto, Y. Matsushima, N. Takai, A. Kikuchi, Y. Sakurai, T. Okano, Analytical Chemistry, 68, 100-105 (1996) H. Kanazawa, Y. Kashiwase, K. Yamamoto, Y. Matsushima, A. Kikuchi, Y. Sakurai, T. Okano, Analytical Chemistry, 69, 823-830 (1997)

  In order to introduce a functional group into the temperature-responsive polymer, copolymerization with a comonomer having a functional group such as acrylic acid is generally performed. However, in a linear polymer or hydrogel (that is, a cross-linked product), when the introduction rate of acrylic acid is increased, the phase transition temperature rises remarkably, and the transition behavior becomes insensitive. It was inevitable. That is, until now, there has been no method for introducing many functional groups while maintaining a sensitive temperature responsiveness in a temperature-responsive polymer or hydrogel.

  The present invention relates to a monomer for preparing a polymer or hydrogel (that is, a cross-linked product) into which many functional groups are introduced while maintaining a sensitive temperature response, and a polymer or hydrogel (that is, a cross-linked product). The purpose is to provide goods.

  The present inventors made extensive studies based on the above problems. As a result, it has been found that by making the structure very similar to that of the monomer to be copolymerized, the temperature responsiveness retained by the homopolymer can be expressed, and the present invention has been completed.

  That is, the present invention provides the following general formula (I)

(Wherein R ¹ represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms, and R ² and R ³ each independently represents an alkylene group having 1 to 6 carbon atoms, or R ² and R ³ may be combined to form a cycloalkane, where X is a hydrogen atom, amino group, hydroxyl group, halogen atom, carboxyl group or —COOR ⁵ (R ⁵ is a straight chain having 1 to 6 carbon atoms). Or a branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.) Y represents an amino group, a hydroxyl group, a halogen atom, a carboxyl group, or —COOR ⁵ (R ⁵ represents 1 to 6 represents a linear or branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, a substituted benzyl group.

  The present invention also provides the following general formula (II)

(Wherein R ¹ represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms, and R ² and R ³ each independently represents an alkylene group having 1 to 6 carbon atoms, or R ² and R ³ may be combined to form a cycloalkane, where X is a hydrogen atom, amino group, hydroxyl group, halogen atom, carboxyl group or —COOR ⁵ (R ⁵ is a straight chain having 1 to 6 carbon atoms). Or a branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.) Y represents an amino group, a hydroxyl group, a halogen atom, a carboxyl group, or —COOR ⁵ (R ⁵ represents 1 to 6 represents a linear or branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.)).

  Furthermore, the present invention relates to a repeating unit represented by the general formula (II) and a general formula (III)

(Wherein R ¹ represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms, R ⁵ represents a linear or branched alkyl group having 1 to ⁶ carbon atoms, and R ⁶ represents a carbon number. 1 to 6 represents a linear or branched alkyl group.).

  In addition, the present invention provides a cross-linked product containing the polymer and a cross-linked product containing the copolymer.

FIG. 1 is a graph showing the equilibrium swelling degree of each cross-linked product in a phosphate buffer solution at pH 6.4. FIG. 2 is a graph showing the equilibrium swelling degree of each cross-linked product in a phosphate buffer at pH 7.4. FIG. 3 is a graph showing the equilibrium swelling degree of each cross-linked product in a phosphate buffer at pH 9.0.

  In the present invention, examples of the linear or branched alkyl group having 1 to 6 carbon atoms include methyl group, ethyl group, n-propyl group, i-propyl group, n-butyl group, i-butyl group, s- And butyl, t-butyl, n-pentyl and n-hexyl groups).

  In the present invention, examples of the alkylene group having 1 to 6 carbon atoms include a monomethylene group, a dimethylene group, a trimethylene group, a tetramethylene group, a pentamethylene group, and a hexamethylene group. A preferred methylene group is a monomethylene group.

In the present invention, the halogen atom means any of a fluorine atom, a chlorine atom, a bromine atom and an iodine atom. A particularly preferred halogen atom is a chlorine atom.
In the present invention, the substituent of the substituted phenyl group is not particularly limited, and examples thereof include a nitro group and a methyl mercapto group. A particularly preferred substituent of the substituted phenyl group is a nitro group.

In the present invention, the substituted benzyl group is not particularly limited, and examples thereof include a triphenylmethyl group and a diphenylmethyl group.
In general formula (I) and general formula (II), when R ² and R ³ are combined to form a ring, preferred rings include cyclopropane, cyclobutane, cyclopentane, cyclohexane, cyclooctane and the like. .

  In the present invention, the degree of polymerization of the polymer composed of the repeating unit represented by formula (II) is not particularly limited as long as the number of repeating units is 2 or more, but is preferably 50 to 500.

  In the present invention, the copolymer composed of the repeating unit represented by the general formula (II) and the repeating unit represented by the general formula (III) has a total number of repeating units of 2 or more. Although it does not specifically limit, Preferably it is 50-500. Further, the ratio between the former repeating unit and the latter repeating unit is not particularly limited, but the number of repeating units represented by the general formula (II): the number of repeating units represented by the general formula (III) = 1 to 1. 50: 99-50 is preferable.

Hereinafter, the production method of the acrylamide derivative of the present invention, a polymer, a copolymer and a crosslinked product obtained using the derivative will be shown.
The acrylamide derivative of the present invention includes, for example, the general formula (IV)

Wherein R ² is a linear or branched alkyl group having 1 to 4 carbon atoms, R ³ is an alkylene group having 1 to 6 carbon atoms, X is a hydroxyl group, a halogen atom, a carboxyl group, —COOR ⁴ (R ⁴ is Represents a linear or branched alkyl group having 1 to 6 carbon atoms, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group), and Y represents an amino group, a hydroxyl group, a halogen atom, a carboxyl group, or —COOR ⁵ ( R ⁵ represents a linear or branched alkyl group having 1 to 6 carbon atoms, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.) —NH—R ⁵ (R ⁵ represents C1-C6 linear or branched alkyl group, alkyloxycarbonyl group, phenyl group, substituted phenyl group, benzyl group, substituted benzyl group, benzyloxycarbonyl group, substituted benzyl Represents an oxycarbonyl group.), And R ² and R ³ may form a cycloalkane together to form a general formula (V).

(Wherein R ¹ represents a hydrogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms) and can be easily obtained.
Examples of the compound represented by the compound (V) include acrylic acid, methacrylic acid, α-ethylacrylic acid, α-n-butylacrylic acid, α-n-hexylacrylic acid, and those that are generally commercially available is there. This reaction proceeds more efficiently by using a condensing agent, which is dicyclohexylcarbodiimide, diisopropylcarbodiimide, N-ethyl-N′-3-dimethylaminopropylcarbodiimide, benzotriazol-1-yl-tris (dimethylamino). ) Phosphonium hexafluorophosphide diphenylphosphoryl azide and the like can be exemplified and used alone or in combination with N-hydroxysuccinimide, 1-hydroxybenzotriazole and the like. The reaction is preferably carried out in a solvent, and any solvent may be used as long as it dissolves the compound but does not participate in the reaction. Water, chloroform, dichloromethane, dichloroethane, benzene, toluene, xylene, acetone, methyl ethyl ketone, tetrahydrofuran , Dioxane, ethyl acetate, acetonitrile, dimethyl sulfoxide, N, N-dimethylformamide, N, N-dimethylacetamide and the like can be used, but are not limited thereto. The reaction normally proceeds smoothly in the range of 0 ° C to 100 ° C.

  When these condensing agents are not used, the reaction proceeds easily by using a compound obtained by converting the general formula (V) into an acid halide. Examples of the acid halide include acrylic acid chloride, methacrylic acid chloride, acrylic acid bromide, and methacrylic acid bromide, some of which are commercially available. Others can be converted to acid halides by a known method using thionyl chloride, phosphorus trichloride, phosphorus pentoxide or the like. The reaction proceeds easily in an organic solvent under an inert gas atmosphere. Any solvent may be used as long as it does not relate to the reaction. For example, chloroform, dichloromethane, dichloroethane, benzene, toluene, xylene, acetone, methyl ethyl ketone, tetrahydrofuran, dioxane, ethyl acetate, acetonitrile, dimethyl sulfoxide, N, N -Dimethylformamide, N, N-dimethylacetamide and the like can be used, but are not limited thereto.

  This reaction is preferably performed in the presence of a basic substance, such as lithium hydroxide, potassium hydroxide, sodium hydroxide, aluminum hydroxide, potassium carbonate, sodium carbonate, potassium acetate, sodium acetate, sodium phosphate, sodium hydride, Calcium hydride, pyridine, triethylamine, N, N-dimethylaniline and the like are preferably used.

  Compounds used in the general formula (IV) include 1- (hydroxymethyl) ethylamine, 1- (hydroxymethyl) propylamine, β-aminobutyric acid, β-aminobutyric acid methyl ester, β-aminobutyric acid p-nitrophenyl ester , Β-aminobutyric acid benzyl ester, 2-aminopropyl chloride, 2-aminopropyl bromide, γ-aminovaleric acid, γ-aminovaleric acid methyl ester, γ-aminovaleric acid p-nitrophenyl ester, γ-aminovaleric acid Benzyl ester, β-aminovaleric acid, β-aminovaleric acid benzyl ester, 1-methylamino-2-aminopropane, 1- (t-butoxycarbonyl) amino-2-aminopropane, 1-benzylamino-2-amino Examples of propane and 1- (benzyloxycarbonyl) amino-2-aminopropane it can. Some of these are commercially available.

  The acrylamide derivative represented by the general formula (I) of the present invention is obtained by homopolymerization, and a polymer comprising a repeating unit represented by the general formula (II) is obtained by a general polymerization method. It can be easily manufactured. In the radical polymerization method, known methods such as bulk polymerization, solution polymerization, and emulsion polymerization can be used, and the polymerization is efficiently started by adding a radical initiator. Examples of the radical initiator suitably used for the reaction include inorganic peroxides such as potassium persulfate and ammonium persulfate. N, N, N ′, N′-tetramethylethylenediamine, which is called a polymerization accelerator, N, N -Rapid polymerization at low temperatures is possible by using in combination with an amine compound such as dimethylparatoluidine. Furthermore, organic peroxides such as dilauroyl peroxide, benzoyl peroxide, di-t-butyl peroxide, t-butyl hydroperoxide, cumene hydroperoxide, and α, α-azobisisobutyronitrile and azobiscyclohexanecarbonitrile. Examples of such azo compounds can be given.

  Examples of the solvent that can be used for the radical polymerization reaction in this case include water, methanol, ethanol, normal propanol, isopropanol, 1-butanol, isobutanol, hexanol, benzene, toluene, xylene, chlorobenzene, dichloromethane, chloroform, and carbon tetrachloride. , Acetone, methyl ethyl ketone, tetrahydrofuran, dioxane, acetonitrile, dimethyl sulfoxide, N, N-dimethylformamide, N, N-dimethylacetamide, and the like, but are not limited thereto. The reaction normally proceeds smoothly in the range of 0 ° C to 100 ° C.

  Copolymers composed of the repeating unit represented by the general formula (II) and the repeating unit represented by the general formula (III) are, for example, isopropylacrylamide, isobutylacrylamide, 3-pentylacrylamide, cyclopentylacrylamide, and 3-hexyl. It can be easily synthesized by copolymerizing acrylamide or the like with a compound represented by the general formula (I). At this time, a general polymerization method is used, and in particular, it can be easily produced by a radical polymerization method. In the radical polymerization method, known methods such as bulk polymerization, solution polymerization, and emulsion polymerization can be used, and the polymerization is efficiently started by adding a radical initiator. Examples of the radical initiator suitably used for the reaction include inorganic peroxides such as potassium persulfate and ammonium persulfate. N, N, N ′, N′-tetramethylethylenediamine, which is called a polymerization accelerator, N, N -Rapid polymerization at low temperatures is possible by using in combination with an amine compound such as dimethylparatoluidine. Furthermore, organic peroxides such as dilauroyl peroxide, benzoyl peroxide, di-t-butyl peroxide, t-butyl hydroperoxide, cumene hydroperoxide, and α, α-azobisisobutyronitrile and azobiscyclohexanecarbonitrile. Examples of the azo compound are not limited to these.

  Examples of the solvent that can be used for the radical polymerization reaction in this case include water, methanol, ethanol, normal propanol, isopropanol, 1-butanol, isobutanol, hexanol, benzene, toluene, xylene, chlorobenzene, dichloromethane, chloroform, and carbon tetrachloride. , Acetone, methyl ethyl ketone, tetrahydrofuran, dioxane, acetonitrile, dimethyl sulfoxide, N, N-dimethylformamide, N, N-dimethylacetamide, and the like, but are not limited thereto. The reaction normally proceeds smoothly in the range of 0 ° C to 100 ° C.

  The cross-linked product containing the repeating unit represented by the general formula (II) and the cross-linked product containing the copolymer represented by the general formula (II) and the general formula (III) are generally polyfunctional compounds called crosslinking agents. Can be easily obtained by carrying out the polymerization reaction in the presence of. Examples of the cross-linking agent include methylene bisacrylamide, ethylene glycol dimethacrylate, glycerin triacrylate, glycerin trimethacrylate, trimethylolpropane triacrylate, pentaerythritol tetraacrylate, etc., some of which are commercially available. At this time, a general polymerization method is used, and in particular, it can be easily produced by a radical polymerization method. In the radical polymerization method, known methods such as bulk polymerization, solution polymerization, and emulsion polymerization can be used, and the polymerization is efficiently started by adding a radical initiator. Examples of the radical initiator suitably used for the reaction include inorganic peroxides such as potassium persulfate and ammonium persulfate. N, N, N ′, N′-tetramethylethylenediamine, which is called a polymerization accelerator, N, N -Rapid polymerization at low temperatures is possible by using in combination with an amine compound such as dimethylparatoluidine. Furthermore, organic peroxides such as dilauroyl peroxide, benzoyl peroxide, di-t-butyl peroxide, t-butyl hydroperoxide, cumene hydroperoxide, and α, α-azobisisobutyronitrile and azobiscyclohexanecarbonitrile. Examples of the azo compound are not limited to these examples.

  Examples of the solvent that can be used for the radical polymerization reaction in this case include water, methanol, ethanol, normal propanol, isopropanol, 1-butanol, isobutanol, n-hexanol, benzene, toluene, xylene, chlorobenzene, dichloromethane, chloroform, and four. Carbon chloride, acetone, methyl ethyl ketone, tetrahydrofuran, dioxane, acetonitrile, dimethyl sulfoxide, N, N-dimethylformamide, N, N-dimethylacetamide and the like can be used, but are not limited thereto. The reaction normally proceeds smoothly in the range of 0 ° C to 100 ° C.

  The temperature responsiveness of the polymer obtained in the present invention is determined by measuring the turbidity change when the temperature of the solution is changed at a constant rate after being dissolved in water or a buffer solution. It can be evaluated by measuring the temperature range required for the operation. In addition, the cross-linked product can be evaluated by measuring the degree of equilibrium swelling at each temperature when the temperature is changed, and evaluating the degree of change.

  Hereinafter, the present invention will be described in more detail with reference to Examples, Comparative Examples, and Reference Examples. However, it is needless to say that the present invention is not limited to these.

[Reference Examples 1 to 4]
N-isopropylacrylamide (abbreviated as IPAAm), acrylic acid (abbreviated as AAc), and 11.5 mg of polymerization initiator azobisisobutyronitrile (abbreviated as AIBN) in the amounts shown in Table 1 were dissolved in 35 ml of tetrahydrofuran. After deaeration and sealing, the mixture was stirred at 65 ° C. for 2.5 hours. After the reaction, precipitation into diethyl ether was repeated twice for purification.

[Reference Examples 5 to 8]
In the examiner, the amounts of N-isopropylacrylamide (abbreviated as IPAAm), acrylic acid (abbreviated as AAc), 26.6 mg of the cross-linking agent methylenebisacrylamide (abbreviated as MBAAm) and 48 μl of the polymerization accelerator N described in Table 2. , N, N ′, N′-tetramethylethylenediamine was dissolved in 10 ml of water. After bubbling nitrogen gas, 200 μl of 40 mg / ml ammonium persulfate aqueous solution was immediately added, and the solution was sucked into the capillary tube. After maintaining at 0 ° C. for 24 hours, the crosslinked product prepared from the capillary was taken out and purified by dipping in cold water for 1 day.

[Example 1]

According to the synthesis scheme of [Chemical 6], 18.1 g of β-aminobutyric acid, 87.5 ml of benzyl alcohol and 40.0 g of p-toluenesulfonic acid were first dissolved in 175 ml of benzene and refluxed. The water formed was removed as an azeotrope with benzene. To the reaction solution, 280 ml of diethyl ether and 280 ml of hexane were added to form a precipitate. This precipitate was separated by filtration and recrystallized in a mixed solution of ether / ethanol to form. 53.8 mg of β-aminobutyric acid benzyl ester / p-toluenesulfonate was obtained as white crystals. ¹ H-NMR measurement results confirmed the synthesis of β-aminobutyric acid benzyl ester / p-toluenesulfonate. The ¹ H-NMR spectrum measurement results are shown below. The underlined portion indicates the position of the corresponding proton in the compound. In the NMR data below, “Φ-” represents a phenyl group.
¹ H-NMR δ (DMSO-d ₆ , ppm) 1.20 (d, 3H, CH ₃ CHCH ₂ ), 2.29 (s, 3H, Ф- CH ₃ ), 2.68 (m, 2H, CH ₃ CH ₂ CH ₂ ), 3.54 (m, 1H, CH ₃ CH ₂ CH ₂ ), 5.14 (s, 2H, COO CH ₂ —Ф), 7.11 and 7.48 (m, 4H, p-toluene) hydrogen of the benzene ring of the sulfonate), 7.39 (m, 5H, hydrogens of the benzene ring of the benzyl ester), 7.81 (m, 2H, CH 3 CH (NH 2) CH 2)
Next, 50.0 g of β-aminobutyric acid benzyl ester / p-toluenesulfonate was dispersed in 300 ml of ether containing 54.3 ml of triethylamine. To this solution, 12.7 ml of acrylic acid chloride was slowly added at 0 ° C. and stirred for 2 hours. The ether layer was collected, concentrated, and purified by silica gel column chromatography to obtain 19.59 g of the objective 2- (benzyloxycarbonyl) isopropylacrylamide as white crystals. ^The synthesis was confirmed by ¹ H-NMR measurement and elemental analysis. The results of ¹ H-NMR spectrum measurement and elemental analysis are shown below. The underlined portion indicates the position of the corresponding proton in the compound.
¹ H-NMR δ (DMSO-d ₆ , ppm) 1.11 (d, 3H, CH ₃ CHCH ₂ ), 2.53 (m, 2H, CH ₃ CH CH ₂ ), 4.20 (m, 1H, CH ₃ CH ₂ CH ₂ ), 5.06 (s, 2H, COO CH ₂ —Ф), 5.55, 6.07 and 6.18 (m, 3H, CH═CH ₂ ), 7.35 (m, 5H) , Benzene ring hydrogen ), 8.10 (d, 1H, CO NH )

C ₁₄ H ₁₇ NO ₃ 247.32
Calculated value: C67.93 H6.94 N5.66
Measurement: C67.92 H6.90 N5.73

^{From the} results of ¹ H-NMR spectrum measurement and elemental analysis, the synthesis of the target compound was confirmed.

[Example 2]

According to the synthesis scheme of [Chemical Formula 7], 1.0 g of 2- (benzyloxycarbonyl) isopropylacrylamide synthesized in Example 1 was dispersed in 50 ml of 1N aqueous sodium hydroxide solution and stirred for 1 hour. After removing the deprotected benzyl alcohol with ether, concentrated hydrochloric acid was added until the total solution was pH2. After hydrogen chloride and water were completely distilled off, methanol was added to the residue and stirred for 2 hours. Only the methanol layer was separated and concentrated to obtain the desired 2-carboxyisopropylacrylamide as a transparent viscous liquid. The structure was confirmed using ¹ H-NMR. ¹ H-NMR δ (DMSO-d ₆ , ppm) 1.35 (d, 3H, CH ₃ CHCH ₂ ), 2.38 (m, 2H, CH ₃ CH CH ₂ ), 4.15 (m, 1H, CH ₃ CH ₂ CH ₂ ), 5.55, 6.06 and 6.12 (m, 3H, CH═CH ₂ ), 8.10 (d, 1H, CO NH )
¹ H-NMR spectrum measurement results confirmed the synthesis of the target compound.

[Examples 3 to 4]
35 ml of tetrahydrofuran containing N-isopropylacrylamide (abbreviated as IPAAm), 2-carboxyisopropylacrylamide (abbreviated as CIPAAm), and 11.5 mg of polymerization initiator azobisisobutyronitrile (abbreviated as AIBN) in the amounts shown in Table 3. After being degassed and sealed, the mixture was stirred at 65 ° C. for 2.5 hours. After the reaction, precipitation in diethyl ether was repeated twice for purification.

[Examples 5 to 7]

  In a test tube, 100 μl and 4 of an aqueous solution of N-isopropylacrylamide (abbreviated as IPAAm), 2-carboxyisopropylacrylamide (CIPAAm), 26.6 mg / ml of a crosslinking agent methylenebisacrylamide (abbreviated as MBAAm) shown in Table 4 8 μl of polymerization accelerator N, N, N ′, N′-tetramethylethylenediamine was dissolved in 0.9 ml of water. After bubbling nitrogen gas, 20 μl of 40 mg / ml ammonium persulfate aqueous solution was immediately added, and the solution was sucked into the capillary tube. After maintaining at 0 ° C. for 24 hours, the crosslinked product prepared from the capillary was taken out and purified by dipping in cold water for 1 day.

[Example 8]

According to the scheme of [Chemical Formula 8], 119.4 ml of 1,2-diaminopropane first purified by distillation and 194.9 ml of triethylamine were dissolved in 500 ml of dehydrated diethyl ether. At 0 ° C., 20.0 ml of carbobenzoxyl chloride was slowly added dropwise and stirred at room temperature for 18 hours. The supernatant was collected, washed with water and concentrated. Ethyl acetate was added to precipitate crystals, which were filtered and removed. The filtrate was concentrated and purified by column chromatography to give 9.51 g of 1-benzyloxycarbonylamino-2-amino-propane as a clear colorless viscous liquid. The synthesis was confirmed by the ¹ H-NMR spectrum measurement results shown below.
¹ H-NMR δ (DMSO-d ₆ , ppm) 0.91 (d, 3H, CH ₃ CHCH ₂ ), 2.80 (m, 1H, CH ₃ CH CH ₂ ), 2.85 (m, 2H, CH ₃ CH ₂ CH ₂ ), 5.01 (s, 2H, COO CH ₂ —Ф), 7.35 (m, 5H, hydrogen on the benzene ring)
8.0 g of 1-amino-2- (benzyloxycarbonyl) aminopropane and 8.0 ml of triethylamine were dissolved in 200 ml of diethyl ether, and 3.74 ml of acryloyl chloride was slowly added dropwise at 0 ° C. After stirring for 2 hours, the mixture was concentrated and ethyl acetate was added to the residue to dissolve it. Furthermore, this solution was washed with water and dried over anhydrous sodium sulfate. Purification by column chromatography gave 3.43 g of 2- (benzyloxycarbonyl) aminoisopropylacrylamide as plate crystals. The synthesis was confirmed by the ¹ H-NMR spectrum measurement results shown below.
¹ H-NMR δ (DMSO-d ₆ , ppm) 1.03 (d, 3H, CH ₃ CHCH ₂ ), 3.04 (m, 2H, CH ₃ CH CH ₂ ), 3.92 (m, 1H, CH ₃ CH ₂ CH ₂ ), 5.01 (m, 2H, COO CH ₂ —Ф), 5.57, 6.07 and 6.18 (m, 3H, CH═CH ₂ ), 7.34 (m, 5H , Hydrogen in the benzene ring)

[Example 9]

1.0 g of 2- (benzyloxycarbonyl) aminoisopropylacrylamide synthesized in Example 8 was dispersed in a hydrobromic acid solution and stirred for 2 hours. After thoroughly removing hydrogen bromide and acetic acid, water was added to the residue. After extracting and removing the deprotected benzyl bromide with ether, the aqueous layer was concentrated. Methanol was added to the residue and stirred for 2 hours. Only the methanol layer was separated and concentrated to obtain the desired 2-aminoisopropylacrylamide hydrobromide as a white solid. The structure was confirmed using ¹ H-NMR.
¹ H-NMR δ (DMSO-d ₆ , ppm) 1.35 (d, 3H, CH ₃ CHCH ₂ ), 2.89 (m, 2H, CH ₃ CH CH ₂ ), 4.21 (m, 1H, CH ₃ CH ₂ CH ₂ ), 5.50, 6.06 and 6.12 (m, 3H, CH═CH ₂ ), 8.08 (d, 1H, CONH)
¹ H-NMR spectrum measurement results confirmed the synthesis of the target compound.

[Comparative Examples 1 to 12]
The N-isopropylacrylamide / acrylic acid copolymer or N-isopropylacrylamide homopolymer prepared in Reference Examples 1 to 4 was dissolved in phosphate buffers at various pH so as to be 0.6%. An intermediate temperature between 90 ° C. and 10% transmittance when the temperature was raised from 20 ° C. was defined as the lower critical solution temperature (LCST). Furthermore, this temperature range was used as an index of the sensitivity of phase transition. That is, the smaller this index is, the more sensitive the phase transition occurs in a narrower temperature range. Table 5 shows the measurement results.

[Examples 10 to 15]

  The N-isopropylacrylamide (abbreviated as IPAAm) / 2-carboxyisopropylacrylamide (abbreviated as CIPAAm) copolymer prepared in Examples 3 and 4 was dissolved in a phosphate buffer solution to a concentration of 0.6%. The LCST and phase transition sensitivity indices were evaluated in the same manner as in Comparative Examples 1-12. Table 6 shows the measurement results. The results of Comparative Examples 1, 5, and 9 are also shown in the table for comparison with the PIPAAm homopolymer.

  As apparent from Table 6 and Table 5 of the comparative example, a copolymer composed of 2-carboxyisopropylacrylamide (CIPAAm) having a structure similar to that of N-isopropylacrylamide (IPAAm) obtained in Examples was about 10 mol. % Of the CIPAAm content, the phase transition occurs at almost the same temperature as observed with the IPAAm homopolymer, and the temperature range at that time is very small. It is completely different.

[Comparative Examples 13 to 16]
Equilibrium swelling degree of each of N-isopropylacrylamide (IPAAm) and acrylic acid (AAc) copolymer crosslinked products of Reference Examples 5 to 8 at a temperature of 10 ° C. to 50 ° C. in a phosphate buffer solution of pH 6.4. It was measured. The equilibrium swelling degree here is a value obtained by dividing the diameter (d) when the equilibrium at each temperature is reached by the initial diameter (d ₀ ) corresponding to the capillary diameter when prepared in Reference Examples 5 to 8. It was defined as the cube ((d / d ₀ ) ³ ). The results are shown in FIG.

[Comparative Examples 17 to 20]
Equilibrium swelling degree of each of N-isopropylacrylamide (IPAAm) and acrylic acid (AAc) copolymer cross-linked products of Reference Examples 5 to 8 at a temperature of 10 ° C. to 50 ° C. in a phosphate buffer solution of pH 7.4. It was measured. The equilibrium swelling degree here is the same definition as in Comparative Examples 13-16. The results are shown in FIG.

[Comparative Examples 21-24]
Equilibrium swelling degree of each N-isopropylacrylamide (IPAAm) and acrylic acid (AAc) copolymer crosslinked product of Reference Examples 5 to 8 at each temperature from 10 ° C. to 50 ° C. in a phosphate buffer solution of pH 9.0. It was measured. The equilibrium swelling degree here is the same definition as in Comparative Examples 13-16. The results are shown in FIG.

[Examples 16 to 18]
Equilibrium of each of the N-isopropylacrylamide (IPAAm) and 2-carboxyisopropylacrylamide (CIPAAm) copolymer crosslinked products of Examples 5 to 7 at a temperature of 10 ° C. to 50 ° C. in a phosphate buffer at pH 6.4 The degree of swelling was measured. The equilibrium swelling degree here is the same definition as in Comparative Examples 13-16. The results are shown in FIG.

[Examples 19 to 21]
Equilibrium of each of the N-isopropylacrylamide (IPAAm) and 2-carboxyisopropylacrylamide (CIPAAm) copolymer crosslinked products of Examples 5 to 7 at a temperature of 10 ° C. to 50 ° C. in a phosphate buffer at pH 7.4 The degree of swelling was measured. The equilibrium swelling degree here is the same definition as in Comparative Examples 13-16. The results are shown in FIG.

[Examples 22 to 24]
Equilibrium of each of the N-isopropylacrylamide (IPAAm) and 2-carboxyisopropylacrylamide (CIPAAm) copolymer crosslinked products of Examples 5 to 7 at a temperature of 10 ° C. to 50 ° C. in a phosphate buffer at pH 9.0. The degree of swelling was measured. The equilibrium swelling degree here is the same definition as in Comparative Examples 13-16. The results are shown in FIG.

  1-3, in the cross-linked copolymer of N-isopropylacrylamide and acrylic acid, as the acrylic acid content increases, the temperature at which the volume change completes moves to the high temperature side, and the equilibrium of each temperature The change in swelling degree also tended to be less sensitive. On the other hand, in the copolymer composed of PIPAAm and 2-carboxyisopropylacrylamide (CIPAAm), the volume change was completed at the same temperature as the PIPAAm homopolymer, and the equilibrium swelling degree change was also sharp. That is, a cross-linked product having an extremely sensitive volume change behavior similar to the PIPAAm homopolymer was obtained while having a carboxyl group rich in reactivity.

[The invention's effect]
According to the present invention, a monomer for preparing a polymer or hydrogel (that is, a crosslinked product) into which many functional groups are introduced while maintaining a sensitive temperature response, and a polymer or hydrogel (that is, a crosslinked product). Product) can be provided.

Claims (24)

General formula (I)

(Wherein R ¹ represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms, and R ² and R ³ each independently represents an alkylene group having 1 to 6 carbon atoms, or R ² and R ³ may be combined to form a cycloalkane, where X is a hydrogen atom, amino group, hydroxyl group, halogen atom, carboxyl group or —COOR ⁵ (R ⁵ is a straight chain having 1 to 6 carbon atoms). Or a branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.) Y represents an amino group, a hydroxyl group, a halogen atom, a carboxyl group, or —COOR ⁵ (R ⁵ represents 1 to 6 represents a linear or branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.)). General formula (II)

(Wherein R ¹ represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms, and R ² and R ³ each independently represents an alkylene group having 1 to 6 carbon atoms, or R ² and R ³ may be combined to form a cycloalkane, where X is a hydrogen atom, amino group, hydroxyl group, halogen atom, carboxyl group or —COOR ⁵ (R ⁵ is a straight chain having 1 to 6 carbon atoms). Or a branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.) Y represents an amino group, a hydroxyl group, a halogen atom, a carboxyl group, or —COOR ⁵ (R ⁵ represents 1 to 6 represents a linear or branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.)). The following general formula (II)

(Wherein R ¹ represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms, and R ² and R ³ each independently represents an alkylene group having 1 to 6 carbon atoms, or R ² and R ³ may be combined to form a cycloalkane, where X is a hydrogen atom, amino group, hydroxyl group, halogen atom, carboxyl group or —COOR ⁵ (R ⁵ is a straight chain having 1 to 6 carbon atoms). Or a branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.) Y represents an amino group, a hydroxyl group, a halogen atom, a carboxyl group, or —COOR ⁵ (R ⁵ represents 1 to 6 represents a linear or branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, a substituted benzyl group.) And a repeating unit represented by the following general formula (III):

(Wherein R ¹ represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms, R ⁵ represents a linear or branched alkyl group having 1 to ⁶ carbon atoms, and R ⁶ represents a carbon number. 1 to 6 represents a linear or branched alkyl group.) And a temperature-responsive copolymer.   The temperature-responsive copolymer according to claim 3, wherein the repeating unit represented by formula (III) is N-isopropylacrylamide.   A crosslinked product comprising the temperature-responsive polymer according to claim 2.   A crosslinked product comprising the temperature-responsive copolymer according to claim 3.   The cross-linked product according to claim 6, wherein the repeating unit represented by formula (III) is N-isopropylacrylamide.   The temperature-responsive polymer according to claim 1 or 2, wherein X is a hydrogen atom.   The temperature-responsive copolymer according to claim 3 or 4, wherein X is a hydrogen atom. General formula (I)

(Wherein R ¹ represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms, and R ² and R ³ each independently represents an alkylene group having 1 to 6 carbon atoms, or R ² and R ³ may be combined to form a cycloalkane, where X is a hydrogen atom, amino group, hydroxyl group, halogen atom, carboxyl group or —COOR ⁵ (R ⁵ is a straight chain having 1 to 6 carbon atoms). Or a branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.) Y represents an amino group, a hydroxyl group, a halogen atom, a carboxyl group, or —COOR ⁵ (R ⁵ represents 1 to 6 represents a straight-chain or branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, and a substituted benzyl group.)). How. General formula (II)

(Wherein R ¹ represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms, and R ² and R ³ each independently represents an alkylene group having 1 to 6 carbon atoms, or R ² and R ³ may be combined to form a cycloalkane, where X is a hydrogen atom, amino group, hydroxyl group, halogen atom, carboxyl group or —COOR ⁵ (R ⁵ is a straight chain having 1 to 6 carbon atoms). Or a branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.) Y represents an amino group, a hydroxyl group, a halogen atom, a carboxyl group, or —COOR ⁵ (R ⁵ represents 1 to 6 represents a linear or branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.). The following general formula (II)

(Wherein R ¹ represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms, and R ² and R ³ each independently represents an alkylene group having 1 to 6 carbon atoms, or R ² and R ³ may be combined to form a cycloalkane, where X is a hydrogen atom, amino group, hydroxyl group, halogen atom, carboxyl group or —COOR ⁵ (R ⁵ is a straight chain having 1 to 6 carbon atoms). Or a branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.) Y represents an amino group, a hydroxyl group, a halogen atom, a carboxyl group, or —COOR ⁵ (R ⁵ represents 1 to 6 represents a linear or branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, a substituted benzyl group.) And a repeating unit represented by the following general formula (III):

(Wherein R ¹ represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms, R ⁵ represents a linear or branched alkyl group having 1 to ⁶ carbon atoms, and R ⁶ represents a carbon number. 1 to 6 represents a linear or branched alkyl group), and a method for maintaining the temperature responsiveness of the polymer.   The method according to claim 12, wherein the repeating unit represented by the general formula (III) is N-isopropylacrylamide.   The method according to claim 10 or 11, wherein X is a hydrogen atom.   The method according to claim 12 or 13, wherein X is a hydrogen atom. In the production of a polymer having temperature response, the following general formula (I)

(Wherein R ¹ represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms, and R ² and R ³ each independently represents an alkylene group having 1 to 6 carbon atoms, or R ² and R ³ may be combined to form a cycloalkane, where X is a hydrogen atom, amino group, hydroxyl group, halogen atom, carboxyl group or —COOR ⁵ (R ⁵ is a straight chain having 1 to 6 carbon atoms). Or a branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.) Y represents an amino group, a hydroxyl group, a halogen atom, a carboxyl group, or —COOR ⁵ (R ⁵ represents 1 to 6 represents a linear or branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.)). General formula (II) in the production of polymers with temperature response

(Wherein R ¹ represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms, and R ² and R ³ each independently represents an alkylene group having 1 to 6 carbon atoms, or R ² and R ³ may be combined to form a cycloalkane, where X is a hydrogen atom, amino group, hydroxyl group, halogen atom, carboxyl group or —COOR ⁵ (R ⁵ is a straight chain having 1 to 6 carbon atoms). Or a branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.) Y represents an amino group, a hydroxyl group, a halogen atom, a carboxyl group, or —COOR ⁵ (R ⁵ represents 1 to 6 represents a linear or branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.)). The following general formula (II) in the production of a temperature-responsive copolymer

(Wherein R ¹ represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms, and R ² and R ³ each independently represents an alkylene group having 1 to 6 carbon atoms, or R ² and R ³ may be combined to form a cycloalkane, where X is a hydrogen atom, amino group, hydroxyl group, halogen atom, carboxyl group or —COOR ⁵ (R ⁵ is a straight chain having 1 to 6 carbon atoms). Or a branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, or a substituted benzyl group.) Y represents an amino group, a hydroxyl group, a halogen atom, a carboxyl group, or —COOR ⁵ (R ⁵ represents 1 to 6 represents a linear or branched alkyl group, a phenyl group, a substituted phenyl group, a benzyl group, a substituted benzyl group.) And a repeating unit represented by the following general formula (III):

(Wherein R ¹ represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms, R ⁵ represents a linear or branched alkyl group having 1 to ⁶ carbon atoms, and R ⁶ represents a carbon number. 1 to 6 linear or branched alkyl groups are used.)   The use according to claim 18, wherein the repeating unit represented by formula (III) is N-isopropylacrylamide.   A method for producing a crosslinked product, wherein the polymer having temperature responsiveness according to claim 17 is further crosslinked with a crosslinking agent.   A method for producing a crosslinked product, wherein the copolymer having temperature responsiveness according to claim 18 is further crosslinked with a crosslinking agent.   The method for producing a crosslinked product according to claim 21, wherein the repeating unit represented by the general formula (III) is N-isopropylacrylamide.   The use according to claim 16 or 17, wherein X is a hydrogen atom.   20. Use according to claim 18 or 19, wherein X is a hydrogen atom.

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