Zwitterionic compounds, their production methods and uses

Abstract

The present invention addresses the problem of providing a zwitterion compound that has excellent protein adsoprtion–suppressing action. A zwitterion compound that is a polymer that includes units represented by formula (1) (in which R1 is a hydrogen atom or a methyl group, R2 is OH or O-, X1 is -O- or -N(Q1)-, Q1 is a hydrogen atom or a C1–6 alkyl group, m is an integer from 1 to 12, and n is an integer from 1 to 4).

Description

[Technical field]

[0001] The present invention relates to a zwitterionic compound, its preparation method and use. [Background technology]

[0002] In recent years, advances in medicine have led to the development of many artificial medical materials, such as artificial organs. It is important that artificial medical materials do not induce undesirable biological reactions in the body.

[0003] The living body has a self-defense reaction system that recognizes foreign substances and responds to them. For example, when a blood vessel is damaged and the subendothelial tissue is exposed and comes into contact with blood, a thrombus formation reaction immediately proceeds. This foreign substance recognition reaction of blood is also triggered when it comes into contact with artificial medical materials, and proteins are adsorbed to the surface of the artificial medical material to form a layer, and cells such as platelets are adsorbed to this layer, which stretches and denatures, resulting in a thrombus. For this reason, the surface of artificial medical materials used in the living body needs to have properties that suppress protein adsorption.

[0004] In general, it is believed that the surface of a material that exhibits protein adsorption inhibitory activity must have four properties: (1) be hydrophilic, (2) be electrically neutral, (3) be a hydrogen bond acceptor, and (4) not be a hydrogen bond donor (Non-Patent Document 1).

[0005] Although the mechanism by which proteins adsorb to the surface of materials is not fully understood, it is believed that the main cause is the energetic interactions that occur when a protein approaches an interface. The main interactions between proteins and interfaces include van der Waals forces, electrostatic attraction, and hydrogen bonds.

[0006] On hydrophobic surfaces, there is a large interfacial free energy between the aqueous solution and the protein, and therefore protein adsorption to the interface is promoted to eliminate this.In contrast, (1) on hydrophilic surfaces, the interfacial free energy is small, so protein adsorption is suppressed.

[0007] The electrostatic interaction between the protein and the interface is suppressed if the material surface is (2) electrically neutral, and (3) a hydrogen bond acceptor and (4) a non-hydrogen bond donor, thus suppressing the hydrogen bond interaction.

[0008] Therefore, the surface of a material that has the above four characteristics is thought to have the effect of suppressing protein adsorption by reducing the energetic interactions that occur when a protein approaches the interface.

[0009] Polymers are useful as materials for inhibiting protein adsorption, and polyethylene glycol (PEG) has been used for a long time. Materials with water-soluble PEG chains grafted onto their surfaces can inhibit protein adsorption to the material surface by repelling approaching proteins with the hydrated PEG chains. However, PEG chains are easily degraded by oxidation and are not suitable for long-term stable use in vivo (Non-Patent Document 1). Therefore, there is a demand for materials that are not easily degraded in vivo and have a high protein adsorption inhibitory effect.

[0010] 2-Methacryloyloxyethyl phosphorylcholine (MPC) is a monomer created to meet the above requirements, and as represented by the following formula, is a material that mimics a phospholipid called phosphatidylcholine, which is a component of cell membranes (Non-Patent Document 2). [ka]

[0011] Polymers polymerized using MPC as a monomer have the above four properties due to the zwitterionic group, i.e., the phosphorylcholine (PC) group, and exhibit protein adsorption inhibitory action. [Prior art documents] [Non-patent literature]

[0012] [Non-Patent Document 1] Holmlin, RE et al. Langmuir 2001, 17, pp. 2841-2850 [Non-Patent Document 2] Vaisocherova, H. et al., Biosens. Bioelectron. 2009, 24, 1924-1930 Summary of the Invention [Problem to be solved by the invention]

[0013] There is a demand for a material that has a more excellent protein adsorption inhibitory effect than a polymer polymerized using MPC as a monomer. Furthermore, the present inventors have found that a polymer polymerized using MPC as a monomer has a problem in that a decrease in pH reduces hydrophilicity, adversely affecting the protein adsorption inhibitory effect.

[0014] Therefore, an object of the present invention is to provide a zwitterionic compound that has an excellent protein adsorption inhibitory effect, particularly an excellent protein adsorption inhibitory effect over a wide pH range including the acidic range. [Means for solving the problem]

[0015] The present inventors have conducted intensive studies to solve the above problems, and have found that the protein adsorption inhibitory effect of the resulting polymer is improved by using a monomer having a structure in which a quaternary ammonium cation group and a phosphodiester group are replaced in the PC group of MPC. The present inventors have also found that a polymer obtained by using a monomer having the above structure exhibits high hydrophilicity even when the pH is lowered, and has an excellent protein adsorption inhibitory effect in a wide pH range including the acidic range. Furthermore, the present inventors have found that a polymer obtained by using a monomer having the above structure has excellent compatibility with sites that become locally acidic in the body (e.g., inflammation sites, cancer onset sites). The present inventors have further studied based on these findings and have completed the present invention.

[0016] That is, the present invention includes the following aspects. Section 1. The following formula (1): [ka] [In the formula, R 1 is a hydrogen atom or a methyl group, R 2 is OH or O - and X 1 is -O- or -N(Q 1 )-and Q 1 is a hydrogen atom or C 1-6 is an alkyl group, m is an integer from 1 to 12; n is an integer from 1 to 4. and the proportion of said units exceeds 90 mol % based on all monomer units. Section 2. The following formula (1): [ka] [In the formula, R 1 is a hydrogen atom or a methyl group, R 2 is OH or O - and X 1 is -O- or -N(Q 1 )-and Q 1 is a hydrogen atom or C 1-6 is an alkyl group, m is an integer from 1 to 12; n is an integer from 1 to 4. and the unit represented by the following formula (2): [ka] [In the formula, R 3 is a hydrogen atom or a methyl group, and R 4 is C 3-16 is an alkyl group, or R 3 is -CO-X2 -R 4 or -CH2-CO-X 2 -R 4 And R 4 is C 1-16 is an alkyl group, X 2 is -O- or -N(Q 2 )-and Q 2 is a hydrogen atom or C 1-6 is an alkyl group. A polymer comprising a unit represented by: Section 2a. The following formula (1): [ka] [In the formula, R 1 is a hydrogen atom or a methyl group, R 2 is OH or O - and X 1 is -O- or -N(Q 1 )-and Q 1 is a hydrogen atom or C 1-6 is an alkyl group, m is an integer from 1 to 12; n is an integer from 1 to 4. and other monomer units having an ethylenically unsaturated double bond, wherein the proportion of methyl methacrylate units in all monomer units is 10 mol % or less, or 5 mol % or less. Section 2b. The polymer according to item 2a, which does not contain methyl methacrylate units. Section 2c. Item 2b. The polymer according to item 2b, wherein the proportion of ethyl methacrylate units in all monomer units is 10 mol % or less, or 5 mol % or less. Section 2d. The polymer according to item 2b or 2c, which does not contain ethyl methacrylate units. Section 2e. The polymer according to any one of items 2a to 2d, wherein the molar ratio of the unit represented by formula (1) to the other monomer unit having an ethylenically unsaturated double bond is from 10:90 to 40:60. Section 3. Item 3. The polymer according to item 2, wherein a molar ratio of the unit represented by formula (1) to the unit represented by formula (2) is from 10:90 to 40:60. Section 4. The following formula (1): [ka] [In the formula, R 1 is a hydrogen atom or a methyl group, R 2 is OH or O - and X 1 is -O- or -N(Q 1 )-and Q 1 is a hydrogen atom or C 1-6 is an alkyl group, m is an integer from 1 to 12; n is an integer from 1 to 4. A film or sheet comprising a polymer containing a unit represented by the formula: Section 4a. A film or sheet comprising the polymer according to any one of items 1 to 3 and 2a to 2e. Section 5. The following formula (1): [ka] [In the formula, R 1 is a hydrogen atom or a methyl group, R 2 is OH or O - and X 1 is -O- or -N(Q 1 )-and Q 1 is a hydrogen atom or C 1-6 is an alkyl group, m is an integer from 1 to 12; n is an integer from 1 to 4. A biocompatible material comprising a polymer containing a unit represented by: Section 5a. A biocompatible material comprising the polymer according to any one of items 1 to 3 and 2a to 2e. Section 5b. A method for imparting biocompatibility to a material or improving the biocompatibility of a material, comprising: [ka] [In the formula, R 1 is a hydrogen atom or a methyl group, R 2 is OH or O - and X 1 is -O- or -N(Q 1 )-and Q 1 is a hydrogen atom or C 1-6 is an alkyl group, m is an integer from 1 to 12; n is an integer from 1 to 4. The method includes the step of applying a polymer comprising a unit represented by Section 5c. A method for imparting biocompatibility to a material or improving the biocompatibility of a material, the method comprising the step of applying a polymer according to any one of items 1 to 3 and 2a to 2e to a surface of the material. Section 5d. The method according to paragraph 5b or 5c, wherein the material is a medical material. Section 5e. In order to impart biocompatibility to a material or to improve the biocompatibility of a material, [ka] [In the formula, R 1 is a hydrogen atom or a methyl group, R 2 is OH or O - and X1 is -O- or -N(Q 1 )-and Q 1 is a hydrogen atom or C 1-6 is an alkyl group, m is an integer from 1 to 12; n is an integer from 1 to 4. The use of a polymer comprising a unit represented by: Section 5f. Use of a polymer according to any one of paragraphs 1 to 3 and 2a to 2e for imparting biocompatibility to a material or for improving the biocompatibility of a material. Section 5g. Use under paragraphs 5e or 5f where the material is a medical material. Section 6. Item 5. The biocompatible material according to item 5 or 5a, which is a surface coating material for a medical material. Section 7. A polymer brush comprising a substrate and a polymer layer formed of a plurality of polymer chains, one end of which is fixed to the substrate and the other end of which is a free end, The plurality of polymer chains are represented by the following formula (1): [ka] [In the formula, R 1 is a hydrogen atom or a methyl group, R 2 is OH or O - and X 1 is -O- or -N(Q 1 )-and Q 1 is a hydrogen atom or C 1-6 is an alkyl group, m is an integer from 1 to 12; n is an integer from 1 to 4. A polymer brush comprising a polymer comprising a unit represented by: Section 7a. A polymer brush comprising the polymer according to any one of items 1 to 3 and 2a to 2e. Section 8. The formula: [ka] [In the formula, R 1 is a hydrogen atom or a methyl group, R 2 is OH or O - and X 1 is -O- or -N(Q 1 )-and Q 1 is a hydrogen atom or C 1-6 is an alkyl group, m is an integer from 1 to 12; n is an integer from 1 to 4. A method for producing a compound represented by the following formula: The formula: [ka] (In the formula, R 5 and R 6 are the same or different and are alkyl groups, R 1 , X 1 , m, and n are the same as above. The compound represented by the following formula (5): [ka] (In the formula, R 7 ~R 9 are the same or different and are alkyl groups, Z is a halogen atom. and subsequently reacting with water and / or an alcohol. Section 9. The formula: [ka] [In the formula, R 1 is a hydrogen atom or a methyl group, R 2 is OH or O -and X 1 is -O- or -N(Q 1 )-and Q 1 is a hydrogen atom or C 1-6 is an alkyl group, m is an integer from 1 to 12; n is an integer from 1 to 4. A method for producing a compound represented by the following formula: The formula: [ka] (In the formula, R 1 , X 1 , m, and n are the same as above. with POCl3 or P2O5, followed by hydrolysis. Effect of the Invention

[0017] The zwitterionic compound of the present invention has an excellent protein adsorption inhibitory effect, and can exhibit an excellent protein adsorption inhibitory effect particularly over a wide pH range including the acidic range. [Brief description of the drawings]

[0018] [Figure 1] FIG. 1 shows the results of measuring the Raman spectrum of the polymer brush of the example. [Diagram 2] FIG. 2 shows the results of measuring the contact angle of the polymer brush of the example with water. [Diagram 3] FIG. 3 shows the results of measuring the contact angle of the polymer brush of the example with a buffer. [Figure 4] FIG. 4 is a micrograph showing that the polymer brush of the example has an excellent inhibitory effect on protein adsorption. [Diagram 5] FIG. 5 is a graph quantifying the protein adsorption inhibitory effect of the polymer brushes of the examples. [Figure 6]FIG. 6 is a micrograph showing that the polymer brush of the example has an excellent inhibitory effect on protein adsorption at pH 2.8. [Figure 7] FIG. 7 is a graph quantifying the protein adsorption inhibitory effect of the polymer brushes of the examples at pH 2.8. [Figure 8] FIG. 8 is a micrograph showing that the film of the example has the protein adsorption suppressing effect. [Figure 9] FIG. 9 is a graph quantifying the protein adsorption inhibitory effect of the film of the example. [Figure 10] FIG. 10 is a graph quantifying the protein adsorption inhibitory effect of the film of the example. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] <Zwitterionic compounds> In one embodiment, the zwitterionic compound has the following formula (1): [ka] (In the formula, R 1 , R 2 , X 1 , m, and n are the same as above. The zwitterionic compound may be in the form of a salt (hereinafter referred to as "zwitterionic polymer A").

[0020] X 1 -N(Q 1 )-, then Q 1 C, as shown 1-6 Examples of the alkyl group include a methyl group, an ethyl group, a propyl group (n-propyl group or i-propyl group), and a butyl group (n-butyl group, i-butyl group, s-butyl group, and t-butyl group). 1 is preferably a hydrogen atom or C 1-4 is an alkyl group, more preferably a hydrogen atom or C 1-3 It is an alkyl group.

[0021] X1 is preferably -O- or -N(H)-. 1 When is -N(H)-, it is less susceptible to bond cleavage than -O- and is stable even under harsh conditions such as high temperatures.

[0022] m and n can be appropriately selected depending on the solubility in the solvent, etc. m is preferably an integer of 4 to 12 from the viewpoint of improving the solubility in the organic solvent, and is preferably an integer of 1 to 3 from the viewpoint of improving the water solubility.

[0023] n is preferably an integer of 1 to 3, and more preferably 1 or 2.

[0024] In the zwitterionic polymer A, the proportion of the units represented by formula (1) relative to all monomer units is, for example, 10 mol % or more, preferably 20 mol % or more, more preferably 30 mol % or more, even more preferably 40 mol % or more, still more preferably 50 mol % or more, particularly preferably 60 mol % or more, and particularly preferably 70 mol % or more.

[0025] In order to further enhance the protein adsorption inhibitory effect, the proportion of units represented by formula (1) relative to all monomer units is preferably 80 mol % or more, more preferably 90 mol % or more, even more preferably more than 90 mol %, still more preferably 95 mol % or more, particularly preferably 99 mol % or more, and especially preferably 100 mol % (homopolymer).

[0026] The zwitterionic polymer A may be a copolymer and may further contain other monomer units in addition to the unit represented by formula (1). The other monomers are usually monomers having an ethylenically unsaturated double bond. In the monomer, the number of ethylenically unsaturated double bonds is, for example, 1 to 3, preferably 1 or 2, and more preferably 1.

[0027] Examples of the monomer having an ethylenically unsaturated double bond include unsaturated carboxylic acids or their salts, esters, or amides; vinyl ethers; vinyl esters; styrenesulfonic acid or its salts; and the like.

[0028] Examples of the unsaturated carboxylic acid include (meth)acrylic acid, crotonic acid, maleic acid, maleic anhydride, fumaric acid, and itaconic acid.

[0029] Examples of salts of unsaturated carboxylic acids include alkali metal salts, and specific examples thereof include sodium salts and potassium salts.

[0030] Examples of the ester of an unsaturated carboxylic acid include alkyl esters and hydroxyalkyl esters. Examples of the alkyl ester include C esters such as methyl ester, ethyl ester, propyl ester, and butyl ester. 1-6 Examples of hydroxyalkyl esters include hydroxy C esters such as hydroxyethyl esters. 2-6 Alkyl esters are included.

[0031] The amide of the unsaturated carboxylic acid includes N-substituted amide and N,N-disubstituted amide. Examples of the N-substituted amide include alkylamide, and specific examples thereof include C amides such as methylamide and ethylamide. 1-6 Examples of N,N-disubstituted amides include dialkylamides, and specific examples thereof include diC amides such as dimethylamide, diethylamide, and ethylmethylamide. 1-6 Examples of such amides include alkylamides.

[0032] Examples of vinyl ethers include ethyl vinyl ether, ethylene glycol monovinyl ether, diethylene glycol monovinyl ether, propyl vinyl ether, and cyclohexyl vinyl ether.

[0033] Examples of vinyl esters include vinyl acetate, vinyl propionate, and vinyl butyrate.

[0034] The other monomers may be used alone or in combination of two or more.

[0035] The other monomer is preferably (meth)acrylic acid, maleic acid, fumaric acid, itaconic acid, a salt thereof, an ester, or an amide thereof, for example, at least one selected from the group consisting of an alkyl(meth)acrylate and an alkyl(meth)acrylamide.

[0036] Examples of alkyl (meth)acrylates include C alkyl (meth)acrylates such as methyl (meth)acrylate, ethyl (meth)acrylate, propyl (meth)acrylate, butyl (meth)acrylate, pentyl (meth)acrylate, hexyl (meth)acrylate, heptyl (meth)acrylate, and octyl (meth)acrylate. 1-10 Examples include alkyl (meth)acrylates.

[0037] The alkyl(meth)acrylamide includes N-alkyl(meth)acrylamide and N,N-dialkyl(meth)acrylamide. Examples of N-alkyl(meth)acrylamide include N-methyl(meth)acrylamide, N-ethyl(meth)acrylamide, N-isopropyl(meth)acrylamide, and the like. 1-4 Examples of N,N-dialkyl(meth)acrylamides include N,N-dialkyl(meth)acrylamides such as N,N-dimethyl(meth)acrylamide and N,N-diethyl(meth)acrylamide. 1-4 Examples include alkyl(meth)acrylamides.

[0038] The other monomer unit is preferably represented by the following formula (2): [ka] (In the formula, R 3 , R 4 , and X2 is the same as above) It is a unit expressed as:

[0039] In one embodiment, R 3 is a methyl group, and R 4 is C 3-6 An alkyl group, specifically, a propyl group, a butyl group, a pentyl group, or a hexyl group is preferred.

[0040] In other embodiments, R 3 is -CO-X 2 -R 4 or -CH2-CO-X 2 -R 4 And R 4 is C 1-6 An alkyl group, specifically, a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group, or a hexyl group, is preferred.

[0041] X 2 -N(Q 2 )-, then Q 2 C, as shown 1-6 Examples of the alkyl group include a methyl group, an ethyl group, a propyl group, and a butyl group.

[0042] In one embodiment, the copolymer preferably contains 10 mol% or less or 5 mol% or less of methyl methacrylate units in all monomer units, and preferably does not contain methyl methacrylate units. In this embodiment, the copolymer preferably contains 10 mol% or less or 5 mol% or less of ethyl methacrylate units in all monomer units, and preferably does not contain ethyl methacrylate units.

[0043] The molar ratio of the unit represented by formula (1) to other monomer units (for example, monomer units having an ethylenically unsaturated double bond such as the unit represented by formula (2)) is preferably 10:90 to 40:60, more preferably 10:90 to 35:65, and even more preferably 10:90 to 30:70.

[0044] Depending on its polymerization form, the zwitterionic polymer A may be, for example, a random copolymer, an alternating copolymer, a block copolymer, or a graft copolymer, and the shape of the zwitterionic polymer A may be, for example, linear, comb, star, or ladder.

[0045] Zwitterionic polymer A has an excellent protein adsorption inhibitory effect over a wide pH range, for example, a pH range of 1 to 10. Zwitterionic polymer A can also exhibit an excellent protein adsorption inhibitory effect in an acidic range (for example, pH 1 to 3) that is not suitable for a polymer polymerized using MPC as a monomer.

[0046] <Method of producing zwitterionic compound> The method for producing the zwitterionic polymer A is not particularly limited, but for example, a method for producing the zwitterionic polymer A may be carried out by reacting the zwitterionic polymer A with a compound represented by the following formula (3): [ka] (In the formula, R 1 , R 2 , X 1 , m, and n are the same as above. The method may include a step of polymerizing a compound represented by the formula (zwitterionic monomer):

[0047] In the following description, a "compound represented by formula (N)" (N is a positive integer) is referred to as a "compound (N)".

[0048] (Production method of compound (3)) Compound (3) can be obtained, for example, by the following reaction scheme: [ka] (In the formula, L 1 and L 2 are the same or different and each is a halogen atom; R 1 , R 2 , R 5 ~R 9 , X 1, Z, m, and n are the same as above.

[0049] By producing compound (3) according to the above reaction scheme, compound (3) with high purity can be obtained. The purity of compound (3) is, for example, 90% or more, preferably 95% or more, and more preferably 99% or more. The purity is, 1 H-NMR, 31 It is measured by P-NMR.

[0050] <Process (A1)> Step (A1) is a step of reacting compound (9) with compound (8) to obtain compound (7).

[0051] In compounds (8) and (9), L 1 and L 2 Examples of the halogen atom represented by the formula (I) include a chlorine atom, a bromine atom, and an iodine atom.

[0052] The amount of compound (8) used is usually 0.8 to 2.0 mol, preferably 1.0 to 1.2 mol, per 1 mol of compound (9).

[0053] The reaction in step (A1) is preferably carried out in the presence of a base. Examples of the base include chain amines (e.g., di- or tri-C amines such as triethylamine and N,N-diisopropylethylamine). 1-4 alkylamines), cyclic amines (e.g., 1,4-diazabicyclo[2.2.2]octane, pyridine, N,N-dimethyl-4-aminopyridine, 2,6-lutidine, 2,6-di-tert-butylpyridine), etc. These bases can be used alone or in combination of two or more.

[0054] The amount of the base used is usually 1.1 to 2.0 mol, preferably 1.2 to 1.5 mol, per 1 mol of compound (9).

[0055] The reaction in step (A1) is preferably carried out in a solvent. Examples of the solvent include polar aprotic solvents, and specific examples thereof include haloalkanes such as dichloromethane and dichloroethane, diethyl ether, tetrahydrofuran, acetonitrile, dimethylsulfoxide, dimethylformamide, and hexamethylphosphoric triamide. These solvents can be used alone or in combination of two or more.

[0056] The reaction temperature and reaction time in step (A1) are not particularly limited as long as the reaction proceeds. The reaction temperature is, for example, 0 to 100° C., preferably 5 to 40° C. The reaction time is, for example, 1 to 24 hours, preferably 2 to 12 hours.

[0057] <Process (B1)> Step (B1) is a step of reacting compound (7) with compound (6) to obtain compound (4).

[0058] The amount of compound (6) used is, for example, 0.5 to 2.0 mol, preferably 0.9 to 1.1 mol, and usually about 1 mol, relative to 1 mol of compound (7).

[0059] The reaction in step (B1) is preferably carried out in a solvent. Examples of the solvent include aprotic solvents, and specific examples thereof include nitriles such as acetonitrile, ethers such as diethyl ether and tetrahydrofuran, ethyl acetate, dimethyl sulfoxide, dimethylformamide, hexamethylphosphoric triamide, hexane, benzene, toluene, and xylene. These solvents can be used alone or in combination of two or more.

[0060] The reaction temperature and reaction time in step (B1) are not particularly limited as long as the reaction proceeds. The reaction temperature is, for example, 0 to 100° C., preferably 5 to 40° C. The reaction time is, for example, 1 to 100 hours, preferably 2 to 80 hours.

[0061] <Process (C1)> Step (C1) is a step of reacting compound (4) with compound (5) and subsequently reacting it with water and / or alcohol to obtain compound (3).

[0062] In compound (5), R 7 ~R 9 Examples of the alkyl group represented by the formula (I) include C methyl, ethyl, propyl, and butyl groups. 1-4 Examples of such groups include alkyl groups.

[0063] Examples of the halogen atom represented by Z include a chlorine atom, a bromine atom, and an iodine atom.

[0064] The amount of compound (5) used is usually 2 to 40 mol, preferably 12 to 20 mol, per 1 mol of compound (4).

[0065] The reaction with compound (5) is preferably carried out in a solvent. Examples of the solvent include aprotic solvents, and specific examples thereof include acetonitrile, hexane, benzene, toluene, xylene, etc. These solvents can be used alone or in combination of two or more.

[0066] In the reaction with compound (5), the reaction temperature and reaction time are not particularly limited as long as the reaction proceeds. The reaction temperature is, for example, −10 to 40° C., preferably −5 to 30° C. From the viewpoint of purity, the reaction time is preferably 5 hours or more, more preferably 6 hours or more, even more preferably 7 hours or more, still more preferably 8 hours or more, particularly preferably 9 hours or more, and is usually 20 hours or less, preferably 15 hours or less.

[0067] Examples of alcohol include C such as methanol and ethanol. 1-4 These alcohols may be used alone or in combination of two or more.

[0068] In the reaction with water and / or alcohol, the reaction temperature and reaction time are not particularly limited as long as the reaction proceeds. The reaction temperature is, for example, 0 to 100° C., preferably 5 to 40° C. The reaction time is, for example, 1 to 24 hours, preferably 2 to 12 hours.

[0069] <Process (A2)> Step (A2) is a step of reacting compound (9) with compound (6) to obtain compound (10).

[0070] The amount of compound (6) used is, for example, 0.5 to 2.0 mol, preferably 0.9 to 1.1 mol, and usually about 1 mol, relative to 1 mol of compound (9).

[0071] The reaction in step (A2) is preferably carried out in a solvent, similarly to step (B1).

[0072] The reaction temperature and reaction time in step (A2) are not particularly limited as long as the reaction proceeds. The reaction temperature is, for example, 10 to 150° C., preferably 20 to 120° C. The reaction time is, for example, 1 to 48 hours, preferably 2 to 36 hours.

[0073] <Process (B2)> In one embodiment, step (B2) is a step of reacting compound (10) with POCl 3 , followed by hydrolysis to obtain compound (3).

[0074] The amount of POCl3 used is usually 1 to 100 mol, preferably 8 to 10 mol, per mol of compound (10).

[0075] The reaction with POCl3 may be carried out in the presence of a base. Examples of the base include linear amines (e.g., tri-C such as triethylamine). 1-4 alkylamines), cyclic amines (e.g., pyridine, imidazole), etc. These bases can be used alone or in combination of two or more.

[0076] The amount of the base used is usually 1.2 to 120 mol, preferably 9 to 12 mol, per 1 mol of compound (10).

[0077] The reaction with POCl3 is preferably carried out in a solvent. Examples of the solvent include aprotic solvents, and specific examples thereof include acetonitrile, diethyl ether, tetrahydrofuran, ethyl acetate, dimethyl sulfoxide, dimethylformamide, hexamethylphosphoric triamide, hexane, benzene, toluene, and xylene. These solvents can be used alone or in combination of two or more.

[0078] In the reaction with POCl3, the reaction temperature is, for example, −40 to 5° C., and preferably −30 to 0° C. The reaction time is, for example, 0.5 to 24 hours, and preferably 1 to 12 hours.

[0079] In another embodiment, step (B2) is a step of reacting compound (10) with P2O5, followed by hydrolysis to obtain compound (3).

[0080] The amount of P2O5 used is usually 0.5 to 10 mol, preferably 1 to 5 mol, per 1 mol of compound (10).

[0081] The reaction with P2O5 is preferably carried out in a solvent. Examples of the solvent include aprotic solvents, and specific examples thereof include acetonitrile, diethyl ether, tetrahydrofuran, ethyl acetate, dimethyl sulfoxide, dimethylformamide, hexamethylphosphoric triamide, hexane, benzene, toluene, and xylene. These solvents can be used alone or in combination of two or more.

[0082] In the reaction with P2O5, the reaction temperature is, for example, 10 to 150° C., and preferably 30 to 100° C. The reaction time is, for example, 1 to 12 hours, and preferably 2 to 10 hours.

[0083] Compound (3) and its intermediates may be purified by a conventional method (eg, filtration, chromatography, reprecipitation, concentration, repeating each of these operations two or more times, or a combination of each of these operations).

[0084] (Step of polymerizing compound (3)) The zwitterionic polymer A can be produced by polymerizing the compound (3) obtained in step (C1) or step (B2), etc.

[0085] Compound (3) may be polymerized alone or with other monomers, such as the same monomers described in the section on zwitterionic compounds.

[0086] The molar ratio of compound (3) to other monomers is preferably 1:99 to 99:1, more preferably 5:95 to 95:5, even more preferably 10:90 to 90:10, still more preferably 15:85 to 85:15, and particularly preferably 20:80 to 80:20.

[0087] The polymerization of the compound (3) may be carried out in the presence of a catalyst, a crosslinking agent, a polymerization initiator, etc. Examples of the polymerization initiator include a thermal polymerization initiator and a photopolymerization initiator.

[0088] Examples of the thermal polymerization initiator include peroxides and azo compounds.

[0089] Examples of peroxides include dialkyl peroxides (e.g., di-t-butyl peroxide, dicumyl peroxide), diacyl peroxides (e.g., lauroyl peroxide, benzoyl peroxide), peracids or peracid esters (e.g., t-butyl hydroperoxide, cumene hydroperoxide), and the like.

[0090] Examples of the azo compound include azobisisobutyronitrile (AIBN), 2,2'-azobis(2-methylbutyronitrile), and 2,2'-azobis(2,4-dimethylvaleronitrile).

[0091] The thermal polymerization initiators can be used alone or in combination of two or more.

[0092] Examples of the photopolymerization initiator include benzoins (eg, benzoin, benzoin methyl ether, benzoin ethyl ether, benzoin isopropyl ether), acetophenone, benzophenone, benzoylbenzoic acid, 1,4-dibenzoylbenzene, dibenzoyl, and combinations thereof.

[0093] The photopolymerization initiator may be combined with a photosensitizer, such as trialkylamine, trialkanolamine (e.g., triethanolamine), dialkylaminobenzoic acid alkyl ester (e.g., ethyl dimethylaminobenzoate, amyl dimethylaminobenzoate), dialkylaminobenzophenone (e.g., 4-(dimethylamino)benzophenone, 4,4-bis(diethylamino)benzophenone), or a combination thereof.

[0094] The polymerization of compound (3) may be carried out in the presence of a solvent. Examples of the solvent include protic solvents (e.g., water, methanol, ethanol, propanol, butanol, etc.) and aprotic solvents (e.g., ethyl acetate, dimethyl sulfoxide, dimethylformamide, hexamethylphosphoric triamide, hexane, benzene, toluene, xylene, etc.). These solvents may be used alone or in combination of two or more.

[0095] The polymerization of compound (3) is not particularly limited as long as the polymerization proceeds.

[0096] In the case of thermal polymerization, the polymerization temperature is, for example, 30 to 150° C., and preferably 50 to 100° C. The polymerization time is, for example, 5 to 20 hours, and preferably 10 to 15 hours.

[0097] In the case of photopolymerization, active energy rays (e.g., ultraviolet rays) are usually irradiated. The amount of irradiation is not particularly limited, but is, for example, 50 to 10,000 mJ / cm. 2 It is.

[0098] The polymer of compound (3) may be purified by a conventional method (eg, dialysis).

[0099] The polymer may be in a solid or gel form. The solid polymer may have, for example, a one-dimensional shape (e.g., a chain shape), a two-dimensional shape (e.g., a film shape, a sheet shape), or a three-dimensional shape (e.g., a tube shape).

[0100] The film or sheet-shaped polymer can be obtained, for example, by a method including a step of applying a polymerizable composition containing compound (3) and a solvent to a substrate, and a step of polymerizing the polymerizable composition applied to the substrate.

[0101] Examples of the material of the substrate include glass, metal (e.g., chromium, cobalt, nickel, titanium, and alloys containing these), ceramics (e.g., hydroxyapatite), and resin (e.g., fluorine-based resin, polyolefin-based resin, (meth)acrylic resin, polystyrene-based resin, polyester-based resin, polycarbonate-based resin, cellulose resin, and polyamide-based resin). The substrate may be a releasable substrate.

[0102] Examples of methods for applying the composition to a substrate include coating methods (eg, spin coating, bar coating, gravure coating), dipping, and spraying.

[0103] <Molded body> In one embodiment, the shaped body comprises zwitterionic polymer A. The shaped body is preferably a film or sheet.

[0104] The molded article may further contain additives. Examples of additives include stabilizers (e.g., heat stabilizers, antioxidants, and ultraviolet absorbers), fillers, antistatic agents, flame retardants, surfactants, plasticizers, antifouling agents, and the like. These additives may be used alone or in combination of two or more.

[0105] The thickness of the molded product varies depending on the shape, but in the case of a film, the thickness is, for example, 0.5 to 500 μm, preferably 1 to 200 μm, and in the case of a sheet, the thickness is, for example, 0.5 to 5 mm, preferably 1 to 3 mm.

[0106] The molded article can be produced by molding the zwitterionic polymer A by a conventional molding method such as extrusion molding, injection molding, blow molding, cast molding, calendar molding, inflation molding, etc. For example, when the molded article is a film or sheet, the molded article can be produced by a melt film-forming method (e.g., T-die method, calendar method, inflation method) or a solution film-forming method (e.g., cast method).

[0107] <Polymer brush> In one embodiment, the polymer brush is a polymer brush comprising a substrate and a polymer layer formed of a plurality of polymer chains, one end of which is fixed to the substrate and the other end of which is free, wherein the plurality of polymer chains comprise a zwitterionic polymer A.

[0108] The substrate may be, for example, the same as that described in (Step of polymerizing compound (3)) of <Method for producing zwitterionic compound>. The substrate may be one that has been subjected to a surface treatment (for example, a cleaning treatment or an etching treatment).

[0109] The polymer brushes can be produced, for example, by a method including the steps of (a) fixing functional groups that serve as initiation points of a polymerization reaction to the surface of a substrate, and (b) polymerizing a monomer from the initiation points.

[0110] As the step (a), a method of fixing an initiator containing a functional group that serves as the initiation point of the polymerization reaction (hereinafter referred to as "brush initiator") to the surface of the substrate is preferable. As the method, a method of applying a composition containing the brush initiator and a solvent to the substrate is preferable. As the method of application to the substrate, for example, the same method as the method described as the method of application to the substrate in the method of producing a film-like polymer in <Method of producing a zwitterionic compound> can be adopted.

[0111] As the brush initiator, a compound having a bonding group capable of bonding to the base material and a polymerization initiation group serving as a polymerization initiation point can be preferably used.

[0112] Examples of the binding group include reactive silyl groups such as trimethoxysilyl group and triethoxysilyl group, thiol group, disulfide group, phosphate group, etc. Among these, reactive silyl groups are preferred.

[0113] Examples of the polymerization initiator group include halogenated alkyl groups such as 1-bromoethyl group, 1-methyl-1-bromoethyl group, and chloroethyl group; nitroxy group-containing functional groups such as 2,2,6,6-tetramethylpiperidinyl-1-oxy group, N-(t-butyl)-1-phenyl-2-methylpropylnitroxy group, and N-(t-butyl)-1-diethylphosphono-2,2-dimethylpropylnitroxy group; and SO2Cl group. Among these, halogenated alkyl groups are preferred.

[0114] Examples of brush initiators include trialkoxysilylalkyl haloalkanoates, specific examples of which include those represented by the following formula: [ka] (In the formula, R 10 ~R 12 are each independently an alkoxy group; R 13 is a haloalkyl group, and k is an integer of 2 or greater. Examples of the compound include compounds represented by the following formula:

[0115] R 10 ~R 12 Examples of the alkoxy group represented by the formula (I) include a C alkoxy group such as a methoxy group, an ethoxy group, a propoxy group, and a butoxy group. 1-6 Among these, C 1-4 An alkoxy group is preferred, and a methoxy group or an ethoxy group is more preferred.

[0116] R 13 Examples of the haloalkyl group represented by the formula (I) include haloC groups such as 1-bromoethyl group and 1-methyl-1-bromoethyl group. 1-6 R is an alkyl group. 13 is preferably haloC 1-4 It is an alkyl group.

[0117] The brush initiators may be used alone or in combination of two or more.

[0118] Examples of the solvent include protic solvents (e.g., water, methanol, ethanol, propanol, butanol, etc.) and aprotic solvents (e.g., ethyl acetate, acetone, dimethyl sulfoxide, dimethylformamide, hexamethylphosphoric triamide, hexane, benzene, toluene, xylene, etc.) These solvents can be used alone or in combination of two or more.

[0119] When a composition containing a surface initiator and a solvent is applied to a substrate, the surface of the substrate reacts with the surface initiator to form a bond. The reaction temperature and reaction time are not particularly limited as long as the surface initiator is fixed to the substrate. The reaction temperature is, for example, 10 to 40°C, preferably 15 to 30°C, and the reaction time is, for example, 1 to 36 hours, preferably 5 to 30 hours.

[0120] In step (b), the monomer used is not particularly limited as long as it contains compound (3), and may be compound (3) alone or a combination of compound (3) and another monomer. As the other monomer, for example, the same monomers as those described in the section <Zwitterionic compound> can be used.

[0121] The polymerization method in step (b) is preferably a radical polymerization method, more preferably a living radical polymerization method. Examples of living radical polymerization methods include atom transfer radical polymerization (ATRP), reversible addition / fragmentation chain transfer polymerization (RAFT), and nitroxide-mediated polymerization (NMP). Among these, ATRP and NMP are preferred, and ATRP is preferred from the viewpoint of reaction stability.

[0122] The polymerization is preferably carried out in the presence of a catalyst. The catalyst is usually a metal catalyst, specific examples of which include metal halide catalysts and metal complex catalysts.

[0123] Examples of the metal halide catalyst include copper halide catalysts such as copper chloride (I), copper chloride (II), copper bromide (I), and copper bromide (II); titanium halide catalysts such as titanium chloride (II), titanium chloride (III), titanium chloride (IV), and titanium bromide (IV); iron halide catalysts such as iron chloride (II), iron chloride (III), iron bromide (II), and iron bromide (III); cobalt halide catalysts such as cobalt chloride (II) and cobalt bromide (II); nickel halide catalysts such as nickel chloride (II) and nickel bromide (II); molybdenum halide catalysts such as molybdenum chloride (III) and molybdenum chloride (V); and ruthenium halide catalysts such as ruthenium chloride (III).

[0124] Examples of the metal complex catalyst include ruthenium complex catalysts, iron complex catalysts, nickel complex catalysts, palladium complex catalysts, rhodium complex catalysts, copper complex catalysts, rhenium complex catalysts, and molybdenum complex catalysts.

[0125] The catalyst may be used alone or in combination of two or more. The catalyst is preferably a metal halide catalyst, more preferably a copper halide catalyst.

[0126] The catalyst may be combined with a ligand. The ligand is preferably an electron donating ligand, for example, a polydentate amine such as tris[2-(dimethylamino)ethyl]amine, 1,4,8,11-tetraazacyclotetradecane, 1,4,8,11-tetramethyl-1,4,8,11-tetraazacyclotetradecane, 1,1,4,7,10,10-hexamethyltriethylenetetramine, N,N,N',N",N"-pentamethyldiethylenetriamine; tris[2-pyridylmethyl]amine, N-butyl-2-pyridylmethanimine, N-dodecane, etc. pyridine-based compounds such as N-(2-pyridylmethylene) amine, N-octadecyl-N-(2-pyridylmethylene) amine, N-octyl-2-pyridylmethanimine, 4,4'-dinonyl-2,2'-dipyridyl, 4,4'-di-(t-butyl)-2,2'-dipyridyl, 4,4'-dimethyl-2,2'-dipyridyl, N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine, and 2,2'-bipyridyl; phosphine-based compounds; and cyclopentadiene-based compounds.

[0127] The polymerization may be carried out in the presence of a solvent. Examples of the solvent include protic solvents (e.g., water, methanol, ethanol, propanol, butanol), aprotic solvents (e.g., ethyl acetate, dimethyl sulfoxide, dimethylformamide, hexamethylphosphoric triamide, hexane, benzene, toluene, xylene), etc. These solvents can be used alone or in combination of two or more.

[0128] The polymerization is preferably carried out in the presence of an initiator. Examples of the initiator include bromine-based initiators (e.g., ethyl bromoacetate, methyl bromoacetate, (1-bromoethyl)benzene, ethyl 2-bromoisobutyrate), chlorine-based initiators (e.g., methyl chloroacetate, methyl 2-chloropropionate, 2,2-dichloroacetophenone), etc. These initiators can be used alone or in combination of two or more.

[0129] <Biocompatible materials> Zwitterionic polymer A has excellent biocompatibility, and by applying it to the surface of a material, it is possible to impart biocompatibility to the material or improve the biocompatibility of the material, and the material can be used as a biocompatible material. In one embodiment, the biocompatible material contains zwitterionic polymer A. The term "biocompatible material" refers to a material that has properties to which proteins, cells, etc. have difficulty adhering.

[0130] Examples of biocompatible materials include surface covering materials (or surface coating materials) for medical materials such as artificial blood vessels, catheters, artificial organs, artificial joints, artificial dialysis membranes, artificial skin, artificial bones (joints, etc.), contact lenses, adhesive plasters, and bandages; preservative solutions for contact lenses; surface modifiers for diagnostic medical devices such as biochips and microarray chips; cell culture sheets; preservatives for blood, proteins, and cells; anticoagulants for proteins; moisturizers, pack materials (cosmetics), and the like. EXAMPLES

[0131] The present invention will be described in more detail below based on examples, but the present invention is not limited to these examples.

[0132] [Synthesis example: Zwitterionic monomer] Synthesis Example 1. Synthesis of (2-(methacryloyloxy)ethyldimethylammonio)ethyl phosphate (MCHP) Step A1: Synthesis of diethyl 2-bromoethylphosphate [ka] N,N-Dimethyl-4-aminopyridine (0.18 g, 1.5 mmol) was placed in a flask with a navel under a nitrogen atmosphere. To this was added dehydrated dichloromethane (27 mL), dehydrated triethylamine (3.0 mL, 22 mmol), and 2-bromoethanol (1.0 mL, 15 mmol). Diethyl chlorophosphate (2.9 mL, 20 mmol) was added dropwise to this solution and stirred at 0° C. for 1 hour. The resulting reaction mixture was warmed to room temperature and stirred overnight. The resulting triethylamine hydrochloride was then filtered off, and the filtrate was transferred to a separatory funnel and washed with saturated sodium bicarbonate water. The resulting organic layer was washed with water, saline, and then with 2N HCl. The resulting organic layer was further washed with water and saline, dehydrated over anhydrous magnesium sulfate, filtered, and concentrated to obtain the title compound as a colorless liquid (3.16 g, 81%). 1 H NMR (400 MHz,CDCl3): δ=1.35-1.37(t, J=6.87, 7.16, 6H, CH3-CH2), 3.53-3.56(t, J=6.30, 2H, CH2-Br), 4.12-4.18(quin, J=6.87, 7.16, 4H, PO-CH2-CH3), 4.29-4.33(dt, J=6.30, 2H, Br-CH2-CH2-OP)

[0133] Step B1: Synthesis of (2-(methacryloyloxy)ethyldimethylammonio)ethyl diethylphosphate [ka] Diethyl 2-bromoethylphosphate (1.04 g, 4 mmol) obtained in step A1, 2-(dimethylamino)ethyl methacrylate (0.76 mL, 4.5 mmol), and acetonitrile (7 mL) were stirred at room temperature for 3 days. The solution was concentrated to obtain a crude product as a colorless liquid. The crude product was reprecipitated with ethyl acetate and washed with ethyl acetate to obtain the title compound as a colorless viscous liquid (1.43 g, 64%). 1H NMR (400 MHz,DMSO-d6): δ=1.24-1.28 (dt,J=0.91, 7.02, 6H,CH3CH2-OP), 1.90 (s, 3H,CH3-), 3.19 (s, 6H, -N + (CH3)2-), 3.79-3.83 (m, 4H, CH2-N + (CH3)2-CH2-CH2-OP), 3.83 (dq, 4H, CH3CH2-OP), 4.43-4.67 (q, J=3.4, 6.12, 2H, -N-CH2-CH2-OP), 4.53 (br, 2H, C(O)-O-CH2-), 5.717, 6.162 (t, 2H, C=CH2)

[0134] Step C1: Synthesis of MCHP [ka] (2-(methacryloyloxy)ethyldimethylammonio)ethyl diethylphosphate (0.61 g, 1.5 mmol) obtained in step B1 was placed in a flask with a navel under a nitrogen atmosphere. Dehydrated acetonitrile (15 mL) was added thereto. Bromotrimethylsilane (1.2 mL, 9.0 mmol) was added dropwise to this solution, and the mixture was stirred at 0°C for 9 hours. The solution was concentrated to obtain a crude product as a viscous liquid (yield: 61%). Methanol was added to this crude product, and the mixture was stirred for 9 hours. The product was purified with a strong acid cation exchange resin (H type) (DOWEX 50Wx8 50-100 mesh) along with two washings with a mixed solvent of water and acetone (water:acetone = 9:1) to obtain a colorless liquid. The liquid was then concentrated to remove acetone, and the resulting solution was freeze-dried overnight. Finally, the freeze-dried product was dissolved in a small amount of dehydrated methanol, and the solution was added dropwise to dehydrated acetone under a nitrogen atmosphere to obtain a white powder precipitate. This reprecipitation procedure was repeated three times to obtain the title compound (0.103 g, 27%). 1 H NMR (400 MHz,DMSO-d6): δ=1.90 (s, 3H,CH3-), 3.19 (s, 6H, -N + (CH3)2-), 3.75 (m, 2H, -N+ (CH3)2-CH2-CH2-OP), 3.83 (br, 2H, -O-CH2-CH2-N), 4.27 (br, 2H, -CH2-OP), 4.53 (br, 2H, -O-CH2-), 5.717, 6.162 (t, 2H, C=CH2) (purity 99.89%)

[0135] Synthesis Example 2. Synthesis of MCHP Step A2: Synthesis of 2-cholinium methacrylate bromide [ka] 2-(Dimethylamino)ethyl methacrylate (0.5 mL, 3.0 mmol), 2-bromoethanol (0.2 mL, 3 mmol), and acetonitrile (6 mL) were added to a flask with a navel and replaced with nitrogen, and the mixture was stirred at room temperature for 24 hours. The solution was concentrated to obtain a crude product as a colorless viscous liquid. The crude product was dissolved in chloroform and reprecipitated in ethyl acetate, and filtered and washed with ethyl acetate to obtain a white solid (0.81 g, 96%). 1 H NMR (400 MHz,CD3OD): δ=1.97(s, 3H, CH3-), 3.276(s, 6H, -N + (CH3)2-), 3.613(m, 2H, -N + (CH3)2-CH2-CH2-OH), 3.88(m, 2H, -O-CH2-CH2-), 4.028(m, 2H, -CH2-OH), 4.641(m, 2H, -O-CH2-), 5.717, 6.162(2s, 2H, C=CH2)

[0136] Step B2: Synthesis of MCHP In a three-necked flask purged with nitrogen, 2-cholinium methacrylate bromide (0.34 g, 1.2 mmol) obtained in step A2, imidazole (0.90 g, 12 mmol), and dehydrated acetonitrile (8 mL) were added. To this solution, phosphoryl chloride (0.4 mL, 4.3 mmol) and dehydrated acetonitrile (20 mL) were added dropwise at -25°C. After 20 minutes, water was added and the mixture was left until the next day. The resulting imidazole hydrochloride was filtered off and concentrated under vacuum to obtain the crude product as a white solid (0.624 g). The crude product was passed through DIAION SK104H, which was repeated three times to obtain a yellow oil (0.4976 g, 52%). Furthermore, isolation was performed by reverse-phase silica column chromatography (0.1% by mass TFA aqueous solution:acetonitrile=95:5) to obtain the title compound as a colorless viscous liquid (47.4 mg, 13%). 1 H NMR (400 MHz,DMSO-d6): δ=1.90 (s, 3H,CH3-), 3.19 (s, 6H, -N + (CH3)2-), 3.75 (m, 2H, -N + (CH3)2-CH2-CH2-OP), 3.83 (br, 2H, -O-CH2-CH2-N), 4.27 (br, 2H, -CH2-OP), 4.53 (br, 2H, -O-CH2-), 5.717, 6.162(2s, 2H, C=CH2)

[0137] Synthesis Example 3. Synthesis of MCHP In a nitrogen-purged three-neck flask, the 2-cholinium methacrylate bromide (0.282 g, 1.0 mmol) obtained in step A2 and dehydrated acetonitrile (2.5 mL) were added, and the flask was placed in an ice bath, after which phosphoryl chloride (0.28 mL, 3.0 mmol) was added dropwise. After 12 hours, deionized water (0.28 mL, 16.0 mmol) was added and the mixture was stirred for 12 hours. The solution was concentrated and then dried under reduced pressure to obtain a crude product as a yellow liquid ( 1H NMR yield: 61%). The crude product (0.845 g) was isolated by reverse-phase silica column chromatography (0.1% by mass TFA aqueous solution:acetonitrile = 95:5) to obtain the title compound (0.139 g, 49%).

[0138] Synthesis Example 4. Synthesis of MCHP Step A4: Synthesis of 2-cholinium methacrylate chloride [ka] The 2-cholinium methacrylate bromide obtained in step A2 was dissolved in water, adsorbed onto a strongly acidic cation exchange resin (H type) (DOWEX 50Wx8 50-100 mesh), and eluted with 0.1 M hydrochloric acid. The title compound was obtained by freeze-drying.

[0139] Step B4: Synthesis of MCHP (1) In a flask with a navel under a nitrogen atmosphere, P2O5 (2.896 g, 15 mmol) was placed, and 2-cholinium methacrylate chloride (2.183 g, 10 mmol) obtained in step A4 and dry acetonitrile (15 mL) were placed in the dropping funnel. Dry acetonitrile (50 mL) was added to the flask. (2) The white solid solution was added dropwise to the flask (6 to 7 drops per minute) while adjusting the temperature to 25° C. using an oil bath. After the addition, the reaction was carried out at 70° C. for 5 hours, and the mixture was filtered to separate it into a liquid and a solid. (3) Mili-Q water (1 mL) was added dropwise to the liquid phase, stirred at 70° C. for 3 hours, and freeze-dried to obtain a crude product. (4) The crude product was dissolved in methanol and then dropped into dehydrated acetone under a nitrogen atmosphere to obtain a precipitate. This reprecipitation procedure was repeated three times to obtain a white powder. (5) The obtained white powder was dissolved in a small amount of acetonitrile, and the insoluble matter was removed by filtration. The solution was then added dropwise to dehydrated acetone under a nitrogen atmosphere to obtain the title compound. (5') As an alternative to (5), the obtained white powder was dissolved in ion-exchanged water, adsorbed onto a strongly acidic cation exchange resin (H type) (DOWEX 50Wx8 50-100 mesh), and eluted with an acetic acid solution to obtain the title compound. 1 H NMR (400 MHz, DMSO -d6): δ=1.899 (s, 3H,C H 3-), 3.148 (s, 6H, -N + (C H 3)2-), 3.484 (m, 2H, -N + (CH3)2-C H 2-CH2-OP), 3.760 (m, 2H, -O-CH2-C H 2-N), 4.262 (m, 2H, -C H 2-OP), 4.515 (m, 2H, -OC H 2-), 5.758, 6.081 (s, 2H, C=CH2)(purity 99.97%) 31 P NMR (400 MHz, DMSO-d6): δ=-0.656 (purity 99.93%)

[0140] Synthesis Example 5. Synthesis of 2-{(3-methacrylamidopropyl)dimethylammonio}ethyl phosphate (MACHP) Step A5: Synthesis of N-[3-(dimethylamino)propyl]methacrylamide [ka] Dehydrated dichloromethane was placed in a flask with a navel under a nitrogen atmosphere. Triethylamine (42 mL, 415 mmol) and N,N-dimethyl-1,3-propanediamine (12 g, 143 mmol) were added to this solution. Methacryloyl chloride (9.7 ml, 87 mmol) was added dropwise to this solution and stirred at 0°C overnight. This solution was filtered to remove triethylamine hydrochloride, and the filtrate was concentrated to obtain a crude product as a viscous liquid. This crude product was purified by reduced pressure distillation (5 mmHg, 140°C) to obtain a colorless viscous liquid. This viscous liquid was then subjected to silica chromatography (chloroform:methanol = 100:2 → 100:25) to obtain the title compound as a colorless viscous liquid (8.05 g, 47 mmol, 51%). 1 H NMR (400 MHz, CDCl3):δ=1.53-1.59(m, 2H, -NH-CH2-C H 2-CH2-N(CH3)2), 1.80(s, 3H, C H 3-CH2), 2.11(s, 6H, N(C H 3)2), 2.27-2.30(m, 2H, -NH-CH2-CH2-C H 2-N(CH3)2), 3.23-3.29(m, 2H, -NH-C H 2-CH2--CH2-N(CH3)2), 5.16, 5.61 (2s, 2H, C=CH2), 7.82(s, 1H, N H )

[0141] Step B5: Synthesis of 2-{(3-methacrylamidopropyl)dimethylammonio}ethyl diethyl phosphate [ka] In a round-bottom flask, N-[3-(dimethylamino)propyl]-2-methyl-2-propenamide (3.48 g, 20 mmol) obtained in step A5, diethyl 2-bromoethylphosphate (6.69 g, 26 mmol) obtained in step A1, 5 ml of acetonitrile, and a small amount of polymerization inhibitor 4-methoxyphenol were added. After stirring at room temperature for 24 hours, the solution was concentrated to obtain a crude product as a viscous liquid. This crude product was decanted with 20 times the amount of ethyl acetate, and a viscous liquid was obtained as the title compound (2.71 g, 6.2 mmol, 31%). 1 H NMR (400 MHz, DMSO-d6): δ=1.20-1.23(t, 6H, C H 3-CH2-OP), 1.82(s, 3H, C H 3-CH2), 1.84-1.88(br, 2H, -NH-CH2-C H 2-CH2-N(CH3)2), 3.08 (s, 6H, -N + (C H 3) 2-), 3.12-3.16(m, 2H, -NH-C H2-CH2-CH2-N(CH3)2), 3.35-3.41 (m, 2H, -NH-CH2-CH2-C H 2-N(CH3)2), 3.66-3.68 (m, 2H, -N + (CH3)2-C H 2-CH2-OP), 4.01-4.06(m, 4H, PO-C H 2-CH3), 4.35-39 (br, 2H, CH2-C H 2-OP), 5.28, 5.67 (2s, 2H, C=CH2), 8.02(s, 1H, N H )

[0142] Step C5: Synthesis of MACHP [ka] 2-{(3-methacrylamidopropyl)dimethylammonio}ethyl diethylphosphate (2.71 g, 6.2 mmol) obtained in step B5 was placed in a flask with a navel under a nitrogen atmosphere. Anhydrous acetonitrile (70 mL) was added thereto. Bromotrimethylsilane (10 mL, 65 mmol) was added dropwise to this solution, and the mixture was stirred at 0°C for 9 hours. The solution was concentrated to obtain a crude product as a viscous liquid. The crude product was dissolved in a small amount of anhydrous methanol and added dropwise to anhydrous acetone under a nitrogen atmosphere to obtain the title compound (2.11 g). 1 H NMR (400 MHz, DMSO-d6): δ=1.85 (s, 3H,C H 3-), 1.86-1.90(m, 2H, -NH-CH2-C H 2-CH2-N(CH3)2), 3.09 (s, 6H, -N + (C H 3)2-), 3.24-3.39(m, 2H, -NH-C H 2-CH2-CH2-N(CH3)2), 3.60-3.63 (m, 2H, -NH-CH2-CH2-C H 2-N(CH3)2), 3.73-3.86 (br, 2H, -O-CH2-C H2-N), 4.23 (br, 2H, -C H 2-OP), 5.33, 5.69 (2s, 2H, C=CH2), 8.07(s, 1H, N H )

[0143] Example 1: Zwitterionic polymer Example 1-1: MCHP homopolymer MCHP (0.191 g, 0.68 mmol) and AIBN (1.1 mg, 0.0068 mmol) were dissolved in a DMSO solution (0.68 mL) in a tubular reactor, and degassed by bubbling nitrogen for 30 minutes. Oxygen in the system was removed by freeze degassing and nitrogen purging. Polymerization was carried out at 70°C for 16 hours. The obtained product was purified by cellulose dialysis tube (3500 MWCO) with methanol and deionized water for one day each, and freeze-dried to obtain MCHP polymer (0.0028 g, 15%). 1 H NMR (400 MHz,methanol-d4): δ=1.8-2.0 (br, 4H,-CH2-CH2-), 3.28 (br, 6H, -N + (CH3)2-), 3.74 (br, 2H, -N + (CH3)2-C H 2-CH2-OP), 3.85 (br, 2H, -O-CH2-C H 2-N), 4.31 (br, 2H, -CH2-OP), 4.547 (br, 2H, -O-CH2-)

[0144] Example 1-2: MCHP-BMA (butyl methacrylate) copolymer An MCHP-BMA copolymer (p(MCHP-co-BMA)) was obtained in the same manner as in Example 1-1, except that a monomer mixture of MCHP and BMA in a molar ratio of 40:60 was used instead of MCHP. In the obtained MCHP-BMA copolymer, the molar ratio of MCHP units to BMA units was 13:87. 1 H NMR (400 MHz,methanol-d4): δ=0.959 (br, 3H,C H3-CH2-), 1.42 (br, 2H, CH3-C H 2-CH2-), 1.63 (br, 2H, -CH2-C H 2-CH2-), 3.28 (br, 6H, -N + (CH3)2-), 3.74 (br, 2H, -N + (CH3)2-C H 2-CH2-OP), 3.85 (br, 2H, -O-CH2-C H 2-N), 3.97 (br, 2H, -CH2-CH2-C H 2-O-), 4.31 (br, 2H, -C H 2-OP), 4.48 (br, 2H, -O-CH2-)

[0145] Examples 1-3: MCHP-BMA Copolymers and MCHP-DI (Dimethyl Itaconate) Copolymers Using the same procedure as in Example 1-2, MCHP-BMA copolymers having a molar ratio of MCHP units to BMA units of 22:78 or 29:71, and MCHP-DI copolymers having a molar ratio of MCHP units to DI units of 18:82 (p(MCHP-co-DI)) were produced. Specifically, MCHP (0.6 mmol), AIBN (0.06 mmol), and BMA (2.4 mmol) were dissolved in absolute ethanol (8 mL) in a side-arm flask after N2 exchange, and the temperature was raised to 70 °C for 24 hours to react. The resulting copolymer was dissolved in methanol, purified by a cellulose dialysis tube (3500 MWCO), dissolved in ethanol, and precipitated in hexane to produce an MCHP-BMA copolymer with a molar ratio of MCHP units to BMA units of 22:78. The same procedure was followed as above, except that 0.9 mmol of MCHP and 2.1 mmol of BMA were used, to produce an MCHP-BMA copolymer having a molar ratio of MCHP units to BMA units of 29:71. An MCHP-DI copolymer with a molar ratio of MCHP units to DI units of 18:82 was produced in the same manner as above, except that DI was used instead of BMA and a mixture of deionized water:methanol = 4:6 (volume ratio) was used instead of absolute ethanol. 1 H NMR (500 MHz,methanol-d4): δ= 1.2-1.5 (br, 2H,-C-CH2-C-),3.24 (br, 6H, -N + (CH3)2-), 3.62 (br, 3H, -O-CH3), 3.74 (br, 2H, -N + (CH3)2-CH2-CH2-OP), 4.26 (br, 2H, -CH2-OP), 4.34 (br, 2H, -O-CH2-) As controls, a BMA homopolymer (p(BMA)), an MPC-BMA copolymer (p(MPC-co-BMA)) having a molar ratio of MPC units to BMA units of 29:71, and a DI homopolymer (p(DI)) were produced in the same manner as above.

[0146] [Example 2: Film] A 0.1 g / mL polymer solution (solvent: ethanol) was prepared using the polymers obtained in Examples 1-1 and 1-2, and the polymer solution was cast on a glass substrate and dried at room temperature for one day to produce a film. Also, a 40 mg / mL polymer solution (solvent: ethanol) was prepared using the polymer obtained in Example 1-3, and the polymer solution was cast on a glass substrate and dried at room temperature for one day to produce a film.

[0147] [Example 3: Polymer brush (SI-ATRP)] Glass wafers were cut into 1.0 cm x 2.0 cm pieces, cleaned by sonication in ethanol, water, acetone, and ethanol, successively, for 5 min each, and etched by oxygen plasma for 10 min. (3-Trimethoxysilyl)propyl 2-bromo-2-methylpropionate (TSBM) (0.045 mmol, 0.012 mL) was used as the initiator to obtain a homogeneous initiator monolayer on the glass wafer. 3 mmol of the modifier [MPC (Tokyo Chemical Industry Co., Ltd., trade name "2-methacryloyloxyethyl phosphorylcholine"), MCHP, or MACHP], a glass wafer with a TSBM monolayer, copper (I) bromide (0.12 mmol, 0.017 g), and 2,2'-bipyridyl (0.3 mmol, 0.047 g) were added to a 200 mL navel flask and replaced with a nitrogen atmosphere. Methanol and water (methanol:water = 1:1) as a solvent, followed by ethyl 2-bromoisobutyrate (EBIB) (0.12 mmol, 0.018 mL) were added, and the mixture was shaken at room temperature for 24 hours to obtain MPC-modified glass wafer (Si-p(MPC)), MCHP-modified glass wafer (Si-p(MCHP)), and MACHP-modified glass wafer (Si-p(MACHP)).

[0148] Test Example 1: Raman Spectrum The Raman spectra of the slide glass, the glass wafer with a TSBM monolayer (simply referred to as "TSBM"), the Si-p(MCHP), and the Si-p(MPC) were measured using a laser Raman microscope (Technospex, uRaman-532). The results are shown in Figure 1.

[0149] Si-p(MPC) is 1090cm -1 A peak was observed at 1000 s, which represents the symmetric stretching vibration of OPO in the MPC. For Si-p (MCHP), 917 cm -1 A peak was observed at 1000 nm, which represents the PO stretching vibration of POH in MCHP. In addition, TSBM's 582cm -1 is due to C-Br.

[0150] Test Example 2: Contact angle (WCA) The contact angles of the slide glass (SG), TSBM, Si-p(MPC), Si-p(MCHP), and Si-p(MACHP) with water or buffer were measured using an automatic contact angle meter (Kyowa Interface Science Co., Ltd., DMe-201) as follows. The contact angle with respect to water was measured at room temperature using a 5 μL drop of Mili-Q water. The contact angle for the buffer was measured at room temperature using the buffers shown in the table below, with 5 μL of each buffer being dropped.

[0151] [Table 1]

[0152] The contact angle results for water are shown in FIG. 2, and the contact angle results for buffer are shown in FIG.

[0153] The static water contact angle of the glass slide is about 46.3°, whereas Si-p(MPC) and Si-p(MCHP) are more hydrophilic than the glass slide, with static water contact angles of about 37.8° and 22.7°, respectively, indicating that Si-p(MCHP) has a highly hydrophilic surface under aqueous conditions.

[0154] The contact angles against the buffer solution show that Si-p(MCHP) and Si-p(MACHP) are more hydrophilic and more stable against various pH values ​​than the slide glass, Si-p(TSBM), and Si-p(MPC). For example, as the pH decreases from 3.8 to 1.0, the contact angle of Si-p(MPC) changes significantly (increases), whereas the contact angle of Si-p(MCHP) and Si-p(MACHP) hardly changes.

[0155] Test Example 3: Protein adsorption A solution containing 2 mg / ml of protein (fibrinogen) was prepared in 0.1 M, pH 9.0 sodium carbonate buffer. Separately, FITC (fluorescein isothiocyanate) was dissolved in DMSO to a concentration of 1 mg / mL. While gently stirring the protein solution, 50 μL of FITC solution was added to each 1 mL of protein solution to make 5 mL aliquots. After adding the required amount of FITC solution, the reaction was incubated for 8 hours in the dark at 4 ° C. FITC-labeled fibrinogen PBS solution was obtained by dialysis twice overnight at 0-5 ° C using PBS pH 7.4 and a dialysis membrane (Fujifilm Wako Pure Chemical Industries, Ltd., Dialysis Membrane, size 27). Also, instead of PBS pH 7.4, Mcllvaine buffer pH 2.8 was used, and the same operation as above was performed to obtain FITC-labeled fibrinogen Mcllvaine buffer solution.

[0156] Comparison of Si-p(MPC) and Si-p(MCHP) at pH 7.4 The slide glass, Si-p(MPC), and Si-p(MCHP) were sonicated in PBS at pH 7.4 and incubated overnight at 37°C and 5% CO2. They were incubated in the above FITC-labeled fibrinogen PBS solution for 60 minutes, washed with PBS, and then observed using a fluorescent microscope. The fluorescent microscope photograph is shown in Figure 4(A), and the fluorescence intensity (corresponding to the amount of protein adsorbed) is shown in Figure 5.

[0157] The adsorbed proteins were fixed with 75% alcohol and observed using a scanning electron microscope (SEM). The SEM photograph is shown in Figure 4(B), and the protein adsorption area is shown in Figure 5.

[0158] The amount of protein adsorbed on the slide glass is significantly greater than that on Si-p(MCHP) and Si-p(MPC). The adsorption area of ​​protein on the slide glass is also significantly greater than that on Si-p(MCHP) and Si-p(MPC). Si-p(MCHP) has the least amount of protein adsorption and a low adsorption area of ​​approximately 5%, demonstrating its excellent biocompatibility.

[0159] Comparison of Si-p(MPC) and Si-p(MCHP) at pH 2.8 Glass (Plane), Si-p (MPC), and Si-p (MCHP) were sonicated in pH 2.8 Mcllvaine buffer and incubated overnight at 37°C and 5% CO2. These were incubated for 60 minutes in the above FITC-labeled fibrinogen Mcllvaine buffer solution, washed with PBS, and then observed using a fluorescent microscope. Fluorescent microscope images are shown in Figure 6, and the fluorescence intensity and protein adsorption area are shown in Figure 7.

[0160] Comparison of p(BMA) and p(MCHP-co-BMA) A protein adsorption test was carried out in the same manner as described above for the glass having the p(MCHP-co-BMA) film formed on its surface obtained in Example 2, and for the glass having the p(BMA) film formed on its surface as a control. Fluorescence microscopy photographs are shown in Figure 8, and the fluorescence intensity and protein adsorption area are shown in Figure 9.

[0161] The p(MCHP-co-BMA) film adsorbed significantly less protein than the p(BMA) film, and the protein adsorption area was also smaller, indicating that the film is more biocompatible.

[0162] Comparison between p(MPC-co-BMA) and p(MCHP-co-BMA), and between p(DI) and p(MCHP-co-DI) A protein adsorption test was carried out in the same manner as described above for the glass having the p(MCHP-co-BMA) film formed on its surface and the glass having the p(MCHP-co-DI) film formed on its surface obtained in Example 2, as well as for the glass having the p(BMA) film formed on its surface and the glass having the p(DI) film formed on its surface as controls. The fluorescence intensity and the protein adsorption area are shown in Figure 10.

[0163] The p(MCHP-co-BMA) film has a reduced amount of protein adsorption and an adsorption area compared to the p(MPC-co-BMA) film, and is therefore superior in biocompatibility. In addition, when DI is used instead of BMA as another copolymerized monomer, the amount of protein adsorption and the adsorption area can be reduced, and the biocompatibility is also superior.

Claims

1. The following formula (1): 【Chemistry 1】 [In the formula, R １ is a hydrogen atom or a methyl group, R ２ is OH or O － and X １ is -O- or -N(Q １ )- and Q １ is a hydrogen atom or C １－６ is an alkyl group, m is an integer from 1 to 12, n is an integer from 1 to 4. and the proportion of said units exceeds 90 mol % based on all monomer units.

2. The following formula (1): 【Chemistry 2】 [In the formula, R １ is a hydrogen atom or a methyl group, R ２ is OH or O － and X １ is -O- or -N(Q １ )- and Q １ is a hydrogen atom or C １－６ is an alkyl group, m is an integer from 1 to 12, n is an integer from 1 to 4. and a unit represented by the following formula (2): 【Chemistry 3】 [In the formula, R ３ is a hydrogen atom or a methyl group, and R ４ is C ３－１６ is an alkyl group, or R ３ is -CO-X ２ -R ４ or -CH ２ -CO-X ２ -R ４ And R ４ is C １－１６ is an alkyl group, X ２ is -O- or -N(Q ２ )- and Q ２ is a hydrogen atom or C １－６ is an alkyl group. and the proportion of the unit represented by formula (1) is 10 mol % or more and less than 100 mol % based on all monomer units.

3. 3. The polymer according to claim 2, wherein the molar ratio of the unit represented by formula (1) to the unit represented by formula (2) is from 10:90 to 40:

60.

4. The following formula (1): 【Chemistry 4】 [In the formula, R １ is a hydrogen atom or a methyl group, R ２ is OH or O － and X １ is -O- or -N(Q １ )- and Q １ is a hydrogen atom or C １－６ is an alkyl group, m is an integer from 1 to 12, n is an integer from 1 to 4. wherein the proportion of the units represented by formula (1) is 10 mol % or more based on all monomer units.

5. The following formula (1): 【Chemistry 5】 [In the formula, R １ is a hydrogen atom or a methyl group, R ２ is OH or O － and X １ is -O- or -N(Q １ )- and Q １ is a hydrogen atom or C １－６ is an alkyl group, m is an integer from 1 to 12, n is an integer from 1 to 4. A biocompatible material comprising a polymer containing a unit represented by the formula (1), wherein the proportion of the unit represented by the formula (1) is 10 mol % or more based on all monomer units.

6. The biocompatible material according to claim 5 , which is a surface coating material for a medical material.

7. A polymer brush comprising a substrate and a polymer layer formed of a plurality of polymer chains, one end of which is fixed to the substrate and the other end of which is a free end, The plurality of polymer chains are represented by the following formula (1): 【Chemistry 6】 [In the formula, R １ is a hydrogen atom or a methyl group, R ２ is OH or O － and X １ is -O- or -N(Q １ )- and Q １ is a hydrogen atom or C １－６ is an alkyl group, m is an integer from 1 to 12; n is an integer from 1 to 4. A polymer brush comprising a polymer containing a unit represented by the formula (1), wherein the ratio of the unit represented by the formula (1) is 10 mol % or more based on all monomer units.

8. The following formula: 【Chemistry 7】 [In the formula, R １ is a hydrogen atom or a methyl group, R ２ is OH or O － and X １ is -O- or -N(Q １ )- and Q １ is a hydrogen atom or C １－６ is an alkyl group, m is an integer from 1 to 12, n is an integer from 1 to 4. A method for producing a compound represented by the following formula: The following formula: 【Chemistry 8】 (In the formula, R ５ and R ６ are the same or different and are alkyl groups, R １ , X １ , m, and n are the same as above. The compound represented by the following formula (5): 【Chemistry 9】 (In the formula, R ７ ~R ９ are the same or different and are alkyl groups, Z is a halogen atom. and subsequently reacting with water and / or an alcohol.

9. The following formula: 【Chemistry 10】 [In the formula, R １ is a hydrogen atom or a methyl group, R ２ is OH or O － and X １ is -O- or -N(Q １ )- and Q １ is a hydrogen atom or C １－６ is an alkyl group, m is an integer from 1 to 12; n is an integer from 1 to 4. A method for producing a compound represented by the following formula: The following formula: 【Chemistry 11】 (In the formula, R １ , X １ , m, and n are the same as above. The compound represented by the formula: ３ Or P ２ O ５ followed by hydrolysis.

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