Biological substance adhesion-suppressing material

Abstract

Provided is a coating film having improved fixability to a base body, in particular, improved resistance to elution into a solvent after fixation, and having well-balanced desired properties, while maintaining excellent biological substance adhesion-suppressing performance similar to that of a coating film obtained from a conventional copolymer.　Provided are: a copolymer in which a repeating unit represented by formula (c1) derived from a monomer including a benzophenone skeleton is introduced into a copolymer including a repeating unit represented by formula (a1) derived from an anionic monomer and a repeating unit represented by formula (b1) derived from a cationic monomer; a composition including the copolymer; and a coating film including a cured product thereof.

Description

Biomaterial adhesion inhibitor

[0001] The present invention relates to a novel copolymer, a composition containing the copolymer, a coating film containing a cured product of the copolymer, and a method for producing the same.

[0002] To inhibit the adhesion of biomolecules such as cells, proteins, and sugars, various coating materials with the ability to inhibit the adhesion of biomolecules have been proposed and are applicable to medical or research equipment and instruments.

[0003] For example, phosphorylcholine group-containing polymers are known to have excellent biocompatibility, and various applications as coating materials are being investigated (see, for example, Patent Document 1). The present applicants have also previously discovered that coating materials containing copolymers with specific anionic and cationic groups can form coating films with excellent ability to suppress the adhesion of biomolecules, and that these coating films exhibit excellent adhesion to substrates, and have investigated various applications (see, for example, Patent Document 2).

[0004] International Publication No. 2016 / 140259, International Publication No. 2023 / 080165

[0005] The object of the present invention is to further improve the adhesion to a substrate (particularly the resistance to solvents after adhesion) of a coating agent comprising a copolymer containing specific anionic and cationic groups, while maintaining excellent ability to suppress the adhesion of biomaterials.

[0006] The inventors of the present invention have discovered that by introducing a benzophenone skeleton, which can act as a crosslinking component, into a copolymer containing specific anionic and cationic groups, and curing it by light or radiation irradiation, it is possible to obtain a coating film with a balanced set of desired performance while maintaining excellent ability to suppress the adhesion of biomaterials, and further improving adhesion to the substrate, particularly elution into the solvent after adhesion. This has led to the completion of the present invention.

[0007] The present invention is as follows: [1] Formula (a1) below: (In the formula, T ａ , U ａ１ and U ａ２each independently represent a hydrogen atom, or a linear or branched alkyl group having 1 to 5 carbon atoms; Q ａ represents a single bond, an ester bond or an amide bond; R ａ represents a linear or branched alkylene group having 1 to 10 carbon atoms which may be substituted with a halogen atom; n1 represents an integer of 1 to 10), a repeating unit represented by the following formula (b1): (wherein, T ｂ , U ｂ１ , U ｂ２ and U ｂ３ each independently represent a hydrogen atom, or a linear or branched alkyl group having 1 to 5 carbon atoms; Q ｂ represents a single bond, an ester bond or an amide bond; R ｂ represents a linear or branched alkylene group having 1 to 10 carbon atoms which may be substituted with a halogen atom; An － represents an anion selected from the group consisting of halide ions, inorganic acid ions, hydroxide ions and isothiocyanate ions), a repeating unit represented by the following formula (c1): [wherein, T ｃ represents a hydrogen atom, or a linear or branched alkyl group having 1 to 5 carbon atoms; Q ｃ represents a single bond, an ester bond or an amide bond; R ｃ１ represents a single bond, a linear or branched alkylene group having 1 to 10 carbon atoms which may be substituted with a halogen atom, or a group of the formula: (wherein, Alk is a linear or branched alkylene group having 1 to 5 carbon atoms which may be substituted with a hydroxy group, and n2 is an integer of 1 to 10); R ｃ２ and R ｃ３ each independently represent a hydrogen atom, a hydroxy group, or a linear or branched alkoxy group having 1 to 5 carbon atoms], a copolymer comprising the repeating unit represented by

[0008] [1-1] In the repeating unit represented by formula (a1), T ａ , U ａ１ and U ａ２ are each independently a hydrogen atom or a methyl group; Qａ However, it is an ester bond; R ａ The copolymer according to [1], wherein the group is an ethylene group or a propylene group; n1 is an integer from 1 to 10. [1-2] In a repeating unit represented by formula (b1), T ｂ , U ｂ１ , U ｂ２ and U ｂ３ However, each is independently a hydrogen atom, a methyl group, or an ethyl group; Q ｂ However, it is an ester bond; R ｂ However, it is a methylene group, an ethylene group, or a propylene group; An － However, the copolymer is a halide ion as described in [1] or [1-1]. [1-3] In a repeating unit represented by formula (c1), T ｃ Q ｃ However, it is an ester bond; R ｃ１ However, it is a single bond; R ｃ２ and R ｃ３ However, each is independently a hydrogen atom or a hydroxyl group, the copolymer according to any one of [1] to [1-2].

[0009] [2] Furthermore, the following equation (d1): [In the formula, T ｄ Q represents a hydrogen atom or a linear or branched alkyl group having 1 to 5 carbon atoms; ｄ R represents a single bond, ether bond, or ester bond; ｄ A copolymer according to any one of [1] to [1-3], comprising a repeating unit represented by [wherein the aryl portion is a linear or branched alkyl group having 1 to 18 carbon atoms, a cyclic hydrocarbon group having 3 to 10 carbon atoms, an aryl group having 6 to 10 carbon atoms, an aralkyl group having 7 to 15 carbon atoms, or an aryloxyalkyl group having 7 to 15 carbon atoms (wherein the aryl portion may be substituted with a linear or branched alkyl group having 1 to 5 carbon atoms, which may be substituted with a halogen atom)].

[0010] [2-1] In the repeating unit represented by equation (d1), T ｄ Q ｄ However, it is an ester bond; R ｄHowever, the copolymer is a linear or branched alkyl group having 4 to 18 carbon atoms, as described in either [1] or [2]. [2-2] R ｄ However, the copolymer is a branched alkyl group having 4 to 18 carbon atoms, as described in [2-1].

[0011] [3] The copolymer according to any one of [1] to [2-2], wherein the proportion of repeating units represented by formula (c1) contained in the copolymer is less than 50 mol%. [3-1] The copolymer according to any one of [1] to [3], wherein the proportion of repeating units represented by formula (c1) contained in the copolymer is 5 to 40 mol%. [3-2] The copolymer according to any one of [1] to [3-1], wherein the proportion of repeating units represented by formula (a1) contained in the copolymer is in the range of 10 to 50 mol%.

[0012] [4] A composition comprising the copolymer described in any of [1] to [3-2] and a solvent. [4-1] A composition for forming a coating film comprising the copolymer described in any of [1] to [3-2] and a solvent.

[0013] [5] A coating film comprising a cured copolymer according to any of [1] to [3-2]. [6] The coating film according to [5] having the ability to suppress the adhesion of biological substances.

[0014] [7] A method for producing a coating film, comprising the steps of applying the composition described in [4] or [4-1] to the surface of a substrate, and curing the coated surface. [8] The method for producing the coating film according to [7], wherein curing is carried out by irradiating the coated surface with light or radiation. [8-1] The method for producing the coating film according to [7] or [8], wherein curing is carried out by ultraviolet irradiation, gamma ray irradiation or electron beam irradiation. [8-2] The method for producing the coating film according to any one of [7] to [8-1], further comprising the step of washing the cured product with an aqueous alcohol solvent.

[0015] [9] A container comprising the coating film described in [5] or [6].

[10] The container described in [9] for cell culture or storage of biomaterials. [10-1] The container described in [9] or

[10] is a dish, flask, bag, plate, tube, tray, or bottle for cell culture or storage of biomaterials. [10-2] The container described in

[10] or [10-1] is a biopharmaceutical selected from the group consisting of enzymes, blood coagulation fibrinolytic factors, serum proteins, hormones, vaccines, interferons, erythropoietins, cytokines, toxins, antibodies, antibody-drug conjugates, and fusion proteins.

[11] A flow channel comprising the coating film described in [5] or [6]. [11-1] A flow channel comprising the coating film described in [5] or [6].

[12] A filter comprising the coating film described in [5] or [6]. [12-1] The filter according to

[12] , which is a depth filter, a membrane filter, an ultrafiltration filter, a hollow fiber membrane filter, or a dialysis / reverse osmosis membrane filter.

[0016] The present invention provides a copolymer obtained by introducing a repeating unit represented by formula (c1) derived from a monomer containing a benzophenone skeleton into a copolymer containing a repeating unit represented by formula (a1) derived from an anionic monomer and a repeating unit represented by formula (b1) derived from a cationic monomer, which are known to have excellent ability to suppress the adhesion of biomolecules, and a cured product thereof. A coating film containing such a cured product maintains the same excellent ability to suppress the adhesion of biomolecules as a coating film obtained from conventional copolymers, while further improving adhesion to the substrate, particularly elution into the solvent after adhesion, resulting in a balanced performance of the desired properties. In particular, the coating film of the present invention is typically obtained by a manufacturing method that includes the steps of applying a composition containing the copolymer of the present invention to the surface of a substrate and curing the coated surface. Since curing can be performed not only by light irradiation but also by radiation irradiation, curing is possible even in places where irradiation light cannot reach (for example, inside multilayer filters and dialysis filters). Furthermore, since curing and sterilization can be performed simultaneously by radiation irradiation, a shortening of the process can be expected.

[0017] <Copolymer> In one embodiment, the copolymer of the present invention comprises a repeating unit represented by formula (a1), a repeating unit represented by formula (b1), and a repeating unit represented by formula (c1).

[0018] The repeating unit represented by formula (a1) is a repeating unit containing an anionic group, and is defined by the following formula: (In the formula, T ａ , U ａ１ and U ａ２ Each of these independently represents a hydrogen atom or a linear or branched alkyl group having 1 to 5 carbon atoms; Q ａ R represents a single bond, ester bond, or amide bond; ａ (where n1 represents a linear or branched alkylene group having 1 to 10 carbon atoms, which may be substituted with halogen atoms; n1 represents an integer from 1 to 10).

[0019] In this specification, unless otherwise specified, "~" is used to mean that the numbers before and after it are included as the lower and upper limits.

[0020] In this specification, unless otherwise defined, "linear or branched alkyl group having 1 to 5 carbon atoms" means a monovalent linear or branched saturated aliphatic hydrocarbon having 1 to 5 carbon atoms. Examples include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, s-butyl group, t-butyl group, n-pentyl group, 1-methylbutyl group, 2-methylbutyl group, 3-methylbutyl group, 1,1-dimethylpropyl group, 1,2-dimethylpropyl group, 2,2-dimethylpropyl group, or 1-ethylpropyl group.

[0021] T ａ The group is preferably a hydrogen atom, a methyl group, or an ethyl group, more preferably a hydrogen atom or a methyl group, and particularly preferably a methyl group. ａ１ and U ａ２ The group is preferably a hydrogen atom, a methyl group, or an ethyl group, more preferably a hydrogen atom or a methyl group, and particularly preferably a hydrogen atom.

[0022] In this specification, unless otherwise defined, "ester bond" means -C(=O)-O- or -O-C(=O)-, and "amide bond" means -NHC(=O)- or -C(=O)NH-.

[0023] Q ａ Preferably, the bond is an ester bond (-C(=O)-O-).

[0024] In this specification, unless otherwise defined, “halogen atom” means a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom.

[0025] In this specification, unless otherwise defined, "linear or branched alkylene group having 1 to 10 carbon atoms, which may be substituted with halogen atoms" means a linear or branched alkylene group having 1 to 10 carbon atoms, or a linear or branched alkylene group having 1 to 10 carbon atoms substituted with one or more of the above halogen atoms. Here, "linear or branched alkylene group having 1 to 10 carbon atoms" means a linear or branched divalent group (alkanediyl group) of a saturated aliphatic hydrocarbon having 1 to 10 carbon atoms. Examples of "linear or branched alkylene groups having 1 to 10 carbon atoms" include methylene group, ethylene group, propylene group (propane-1,2-diyl group), trimethylene group, tetramethylene group, 1-methylpropylene group, 2-methylpropylene group, dimethylethylene group, ethylethylene group, pentamethylene group, 1-methyltetramethylene group, 2-methyltetramethylene group, 1,1-dimethyl-trimethylene group, 1,2-dimethyl-trimethylene group, and 2,2-dimethyl-trimethylene group. Examples include ethylene groups, 1-ethyl-trimethylene groups, hexamethylene groups, octamethylene groups, and decamethylene groups. Among these, ethylene groups, propylene groups, trimethylene groups, tetramethylene groups, pentamethylene groups, octamethylene groups, or decamethylene groups are preferred, linear or branched alkylene groups having 1 to 5 carbon atoms, such as ethylene groups, propylene groups, trimethylene groups, tetramethylene groups, and pentamethylene groups, are more preferred, and ethylene groups or propylene groups are particularly preferred. "A linear or branched alkylene group having 1 to 10 carbon atoms substituted with one or more halogen atoms" means that one or more arbitrary hydrogen atoms of the alkylene group are replaced by the halogen atoms. Preferably, a linear or branched alkylene group having 1 to 5 carbon atoms has some or all of its hydrogen atoms replaced by one or more halogen atoms. More preferably, an ethylene group or propylene group has some or all of its hydrogen atoms replaced by one or more halogen atoms. Particularly preferred is an ethylene group or propylene group in which one hydrogen atom is replaced by a chlorine atom (for example, 3-chloropropane-1,2-diyl).

[0026] R ａn1 is preferably a linear or branched alkylene group having 1 to 5 carbon atoms, more preferably a methylene group, an ethylene group, or a propylene group, and particularly preferably an ethylene group or a propylene group.

[0027] The repeating unit represented by formula (b1) is a repeating unit containing a cationic group, and is defined by the following formula: (In the formula, T ｂ , U ｂ１ , U ｂ２ and U ｂ３ Each of these independently represents a hydrogen atom or a linear or branched alkyl group having 1 to 5 carbon atoms; Q ｂ R represents a single bond, ester bond, or amide bond; ｂ An represents a linear or branched alkylene group having 1 to 10 carbon atoms, which may be substituted with halogen atoms; An － (where represents an anion selected from the group consisting of halide ions, inorganic acid ions, hydroxide ions, and isothiocyanate ions)

[0028] T ｂ The group is preferably a hydrogen atom, a methyl group, or an ethyl group, more preferably a hydrogen atom or a methyl group, and particularly preferably a methyl group. ｂ１ , U ｂ２ and U ｂ３ The group is preferably a hydrogen atom, a methyl group, or an ethyl group, more preferably a methyl group or an ethyl group, and particularly preferably a methyl group. ｂ The bond is preferably an ester bond (-C(=O)-O-). ｂ Preferably, it is a linear or branched alkylene group having 1 to 5 carbon atoms, more preferably a methylene group, an ethylene group, or a propylene group, and particularly preferably a methylene group or an ethylene group.

[0029] In this specification, unless otherwise defined, "halide ion" means fluoride ion, chloride ion, bromide ion, or iodide ion, and "inorganic acid ion" means anion derived from an inorganic acid. Examples of inorganic acid ions include carbonate ion, sulfate ion, phosphate ion, hydrogen phosphate ion, dihydrogen phosphate ion, nitrate ion, perchlorate ion, or borate ion.

[0030] An － The ions are preferably halide ions, sulfate ions, phosphate ions, hydroxide ions, and isothiocyanate ions, more preferably halide ions, and particularly preferably chloride ions or bromide ions.

[0031] The repeating unit represented by formula (c1) is a repeating unit containing a benzophenone skeleton, and is defined by the following formula: [In the formula, T ｃ Q represents a hydrogen atom or a linear or branched alkyl group having 1 to 5 carbon atoms; ｃ R represents a single bond, ester bond, or amide bond; ｃ１ This includes single bonds, linear or branched alkylene groups having 1 to 10 carbon atoms which may be substituted with halogen atoms, or formula: (Here, Alk represents a linear or branched alkylene group having 1 to 5 carbon atoms, which may be substituted with a hydroxyl group, and n2 is an integer from 1 to 10) the group; R ｃ２ and R ｃ３ Each of these independently represents a hydrogen atom, a hydroxyl group, or a linear or branched alkoxy group having 1 to 5 carbon atoms.

[0032] T ｃ The group is preferably a hydrogen atom, a methyl group, or an ethyl group, more preferably a hydrogen atom or a methyl group, and particularly preferably a methyl group. ｃ Preferably, the bond is an ester bond (-C(=O)-O-).

[0033] In this specification, unless otherwise defined, "linear or branched alkylene group having 1 to 5 carbon atoms that may be substituted with a hydroxyl group" means a linear or branched alkylene group having 1 to 5 carbon atoms, or a linear or branched alkylene group having 1 to 5 carbon atoms substituted with one or more hydroxyl groups. Examples of "linear or branched alkylene group having 1 to 5 carbon atoms" are as described above, with methylene, ethylene, propylene, or trimethylene groups being preferred. "Linear or branched alkylene group having 1 to 5 carbon atoms substituted with one or more hydroxyl groups" means an alkylene group in which one or more arbitrary hydrogen atoms are replaced by a hydroxyl group, and in particular, an ethylene, propylene, or trimethylene group in which one or two hydrogen atoms are replaced by a hydroxyl group (for example, 2-hydroxypropane-1,3-diyl) is preferred.

[0034] R ｃ１ Preferably, a single bond or formula: (Here, Alk is a methylene group, ethylene group, propylene group or trimethylene group which may be substituted with a hydroxyl group, and n2 is an integer from 1 to 10) and more preferably a single bond, (Here, n² is an integer between 1 and 10), and is particularly preferably a single combination. Also, n² is preferably an integer between 1 and 6, and more preferably an integer between 1 and 5.

[0035] In this specification, unless otherwise defined, "linear or branched alkoxy group having 1 to 5 carbon atoms" means a group -OR (where R is the linear or branched alkyl group having 1 to 5 carbon atoms). Examples include methoxy group, ethoxy group, propoxy group, butoxy group, t-butoxy group, pentyloxy group, etc.

[0036] R ｃ２ and R ｃ３ This is preferably a hydrogen atom or a hydroxyl group, and more preferably a hydrogen atom.

[0037] The proportion (molar ratio) of the repeating units represented by formula (c1) in the copolymer is preferably less than 50 mol%, more preferably in the range of 1 to 50 mol%, even more preferably in the range of 5 to 40 mol%, and particularly preferably in the range of 5 to 30 mol%. There may be two or more types of repeating units represented by formula (c1). In the present invention, it is believed that the benzophenone skeleton portion contained in the repeating units represented by formula (c1) forms crosslinks (i.e., hardens) with other skeleton portions of the copolymer or the substrate, for example, by light irradiation or radiation irradiation, thereby improving adhesion to the substrate and solvent resistance. By setting the proportion (molar ratio) of the repeating units represented by formula (c1) within such a range, such effects can be appropriately achieved.

[0038] The proportion (molar ratio) of the repeating units represented by formula (a1) in the copolymer is, for example, 3 to 80 mol%, preferably 5 to 70 mol%, more preferably 10 to 70 mol%, even more preferably 10 to 60 mol%, and particularly preferably 20 to 50 mol%. There may be two or more types of repeating units represented by formula (a1). Similarly, the proportion (molar ratio) of the repeating units represented by formula (b1) in the copolymer is, for example, 3 to 80 mol%, preferably 5 to 70 mol%, more preferably 10 to 60 mol%, even more preferably 10 to 50 mol%, and particularly preferably 20 to 50 mol%. There may be two or more types of repeating units represented by formula (b1). By setting the repeating units represented by formula (a1) derived from anionic monomers and the repeating units represented by formula (b1) derived from cationic monomers within these ranges, excellent ability to suppress the adhesion of biomolecules can be achieved.

[0039] The copolymer of the present invention is not particularly limited as long as it contains a repeating unit represented by formula (a1), a repeating unit represented by formula (b1), and a repeating unit represented by formula (c1), and may contain other repeating units other than the repeating unit represented by formula (a1) / formula (b1) / formula (c1) as long as it does not impair the object of the present invention. Such a copolymer may contain a total of 40 mol% or more, preferably 50 mol% or more, and more preferably 60 mol% or more of the repeating unit represented by formula (a1) / formula (b1) / formula (c1) as repeating units.

[0040] In another embodiment, the copolymer of the present invention further comprises a repeating unit represented by formula (d1). The repeating unit represented by formula (d1) is a repeating unit comprising a hydrophobic group, and is of the following formula: [In the formula, T ｄ Q represents a hydrogen atom or a linear or branched alkyl group having 1 to 5 carbon atoms; ｄ R represents a single bond, ether bond, or ester bond; ｄ [wherein] represents a linear or branched alkyl group having 1 to 18 carbon atoms, a cyclic hydrocarbon group having 3 to 10 carbon atoms, an aryl group having 6 to 10 carbon atoms, an aralkyl group having 7 to 15 carbon atoms, or an aryloxyalkyl group having 7 to 15 carbon atoms (wherein the aryl portion may be substituted with a linear or branched alkyl group having 1 to 5 carbon atoms, which may be substituted with a halogen atom).

[0041] T ｄ The group is preferably a hydrogen atom, a methyl group, or an ethyl group, more preferably a hydrogen atom or a methyl group, and particularly preferably a methyl group. ｄ Preferably, the bond is an ester bond (-C(=O)-O-).

[0042] In this specification, unless otherwise defined, "linear or branched alkyl groups having 1 to 18 carbon atoms" includes, in addition to the examples of "linear or branched alkyl groups having 1 to 5 carbon atoms" described above, hexyl groups, heptyl groups, octyl groups, nonyl groups, decyl groups, undecyl groups, dodecyl groups, tridecyl groups, tetradecyl groups, pentadecyl groups, hexadecyl groups, heptadecyl groups, or octadecyl groups, or their isomers (e.g., 2-ethylhexyl group).

[0043] In this specification, unless otherwise defined, "cyclic hydrocarbon group having 3 to 10 carbon atoms" means a monovalent group of a monocyclic or polycyclic, saturated or partially unsaturated aliphatic hydrocarbon having 3 to 10 carbon atoms. Among these, monovalent groups of a monocyclic or bicyclic, saturated aliphatic hydrocarbon having 3 to 10 carbon atoms are preferred, and examples include cycloalkyl groups having 3 to 10 carbon atoms such as cyclopropyl, cyclobutyl, or cyclohexyl groups, or bicycloalkyl groups having 4 to 10 carbon atoms such as bicyclo[3.2.1]octyl, bornyl, or isobornyl groups.

[0044] In this specification, unless otherwise defined, "aryl group having 6 to 10 carbon atoms" means a monovalent group of an aromatic hydrocarbon having 6 to 10 carbon atoms, either monocyclic or polycyclic, such as a phenyl group, naphthyl group, or anthryl group. The "aryl group having 6 to 10 carbon atoms" may be substituted with one or more of the following "linear or branched alkyl groups having 1 to 5 carbon atoms, which may be substituted with halogen atoms."

[0045] In this specification, unless otherwise defined, "aralkyl group having 7 to 15 carbon atoms" means the group -R'-R'' (where R' represents the above-mentioned "alkylene group having 1 to 5 carbon atoms" and R'' represents the above-mentioned "aryl group having 6 to 10 carbon atoms"), examples of which include the benzyl group, phenethyl group, or α-methylbenzyl group. The aryl portion of the "aralkyl group having 7 to 15 carbon atoms" may be substituted with one or more of the following "linear or branched alkyl groups having 1 to 5 carbon atoms which may be substituted with halogen atoms".

[0046] In this specification, unless otherwise defined, "aryloxyalkyl group having 7 to 15 carbon atoms" means the group -R'-O-R'' (where R' represents the above-mentioned "alkylene group having 1 to 5 carbon atoms" and R'' represents the above-mentioned "aryl group having 6 to 10 carbon atoms"), examples of which include the phenoxymethyl group, the phenoxyethyl group, or the phenoxypropyl group. The aryl portion of the "aryloxyalkyl group having 7 to 15 carbon atoms" may be substituted with one or more of the following "linear or branched alkyl groups having 1 to 5 carbon atoms which may be substituted with halogen atoms".

[0047] In this specification, unless otherwise defined, "linear or branched alkyl groups having 1 to 5 carbon atoms that may be substituted with halogen atoms" means the linear or branched alkyl groups having 1 to 5 carbon atoms as described above, or the linear or branched alkyl groups having 1 to 5 carbon atoms that are substituted with one or more halogen atoms as described above. Examples of "linear or branched alkyl groups having 1 to 5 carbon atoms" are as described above. On the other hand, "linear or branched alkyl groups having 1 to 5 carbon atoms that are substituted with one or more halogen atoms" means that one or more arbitrary hydrogen atoms of the linear or branched alkyl groups having 1 to 5 carbon atoms as described above are replaced with one or more halogen atoms, and examples include fluoromethyl group, difluoromethyl group, trifluoromethyl group, chlorodifluoromethyl group, chloromethyl group, dichloromethyl group, trichloromethyl group, bromomethyl group, iodomethyl group, 2,2,2-trifluoroethyl group, 2,2,2-trichloroethyl group, perfluoroethyl group, perfluorobutyl group, or perfluoropentyl group.

[0048] R ｄ The group is preferably a linear or branched alkyl group having 4 to 18 carbon atoms, a cyclic hydrocarbon group having 3 to 10 carbon atoms, or an aryl group having 6 to 10 carbon atoms, more preferably a linear or branched alkyl group having 4 to 18 carbon atoms, and particularly preferably an n-butyl group, i-butyl group, t-butyl group, n-pentyl group, i-amyl group, n-hexyl group, n-octyl group, 2-ethylhexyl group, dodecyl group, or stearyl group.

[0049] The proportion (molar ratio) of the repeating units represented by formula (d1) in the copolymer is preferably 0 to 50 mol%, more preferably 0 to 45 mol%, and particularly preferably 10 to 40 mol%. There may be two or more types of repeating units represented by formula (d1). By setting the repeating units represented by formula (d1) within this range, it is useful for maintaining excellent ability to suppress the adhesion of biomaterials and improving adhesion to the substrate.

[0050] The copolymer of the present invention may further include a repeating unit represented by formula (d1) in addition to the repeating unit represented by formula (a1), the repeating unit represented by formula (b1), and the repeating unit represented by formula (c1), and may also include other repeating units other than the repeating unit represented by formula (a1) / formula (b1) / formula (c1) / formula (d1) as long as the object of the present invention is not impaired. Such a copolymer may contain a total of 70 mol% or more, preferably 80 mol% or more, and more preferably 90 mol% or more of the repeating units represented by formula (a1) / formula (b1) / formula (c1) / formula (d1) as repeating units.

[0051] In yet another embodiment, the copolymer of the present invention may further include a crosslinked structure. Examples of such a crosslinked structure include a structural unit derived from a monomer having two or more carbon-carbon unsaturated bonds. Specifically, a monomer having two or more carbon-carbon unsaturated bonds is a polyfunctional monomer having two or more carbon-carbon double bonds, such as a polyfunctional (meth)acrylate compound, a polyfunctional (meth)acrylamide compound, a polyfunctional polyester, or an isoprene compound. The polyfunctional monomer is preferably a difunctional monomer, and more preferably a difunctional (meth)acrylate compound or a difunctional (meth)acrylamide compound. In this specification, unless otherwise defined, "(meth)acrylate compound" means both an acrylate compound and a methacrylate compound. For example, (meth)acrylic acid means both acrylic acid and methacrylic acid. Similarly, "(meth)acrylamide compound" means both an acrylamide compound and a methacrylamide compound.

[0052] As examples of structural units derived from a difunctional (meth)acrylate compound, the following formulae (e1) and (f1): (wherein T ｅ , T ｆ and U ｆ each independently represent a hydrogen atom or a linear or branched alkyl group having 1 to 5 carbon atoms, and R ｅ and R ｆ each independently represent a linear or branched alkylene group having 1 to 10 carbon atoms which may be substituted with a halogen atom; n3 and n4 each independently represent an integer of 1 to 10); and examples include units represented thereby.

[0053] T ｅ and T ｆ are preferably a hydrogen atom, a methyl group or an ethyl group, more preferably a hydrogen atom or a methyl group. U ｆ is preferably a hydrogen atom, a methyl group or an ethyl group, more preferably a hydrogen atom. R ｅ and R ｆ are preferably a linear or branched alkylene group having 1 to 3 carbon atoms which may be substituted with a halogen atom, more preferably an ethylene group or a propylene group, or an ethylene group or a propylene group substituted with one chlorine atom, and particularly preferably an ethylene group or a propylene group. n3 and n4 are preferably an integer of 1 to 6, more preferably an integer of 1 to 5, and particularly preferably 1.

[0054] The proportion (molar ratio) of the crosslinked structure as described above in the copolymer of the present invention is preferably 0 to 40 mol%, more preferably 0 to 30 mol%, and particularly preferably 1 to 30 mol%. The crosslinked structure may be of two or more types. By setting the crosslinked structure within this range, gelation of the solid content during production due to excessive crosslinking can be suppressed, and production can be facilitated.

[0055] The weight-average molecular weight (Mw) of the copolymer of the present invention is typically 10,000 to 1,000,000, preferably 20,000 to 500,000, and more preferably 30,000 to 150,000. The weight-average molecular weight (Mw) can be determined, for example, by Gel Filtration Chromatography (GFC). Further, the copolymer may be any of a random copolymer, a block copolymer, and a graft copolymer, with a random copolymer being preferred.

[0056] <Production Method> In one embodiment, the copolymer of the present invention is obtained, for example, from formula (A): (wherein T ａ , U ａ１ , U ａ２ , Q ａ , R ａ and n1 have the same meanings and preferred embodiments as those in the above formula (a1)), an anionic monomer represented by the formula, and a cationic monomer represented by formula (B): (wherein T ｂ , U ｂ１ , U ｂ２ , U ｂ３ , Q ｂ , R ｂ and An － have the same meanings and preferred embodiments as those in the above formula (b1)), a cationic monomer represented by the formula, and formula (C): (wherein T ｃ , Q ｃ , R ｃ１ , R ｃ２ and R ｃ３ have the same meanings and preferred embodiments as those in the above formula (c1)), the copolymer can be obtained by a production method including a step of polymerizing a monomer mixture containing a monomer containing a benzophenone skeleton represented by the formula.

[0057] As used herein, the term "anionic monomer" means a monomer having an anionic group, and also includes those having a group that can dissociate in water to become anionic. Similarly, the term "cationic monomer" means a monomer having a cationic group, and also includes those having a group that can dissociate in water to become cationic.

[0058] Specific examples of monomers of formula (A) above include vinylphosphonic acid, acid phosphooxyethyl (meth)acrylate, 3-chloro-2-acid phosphooxypropyl (meth)acrylate, acid phosphooxypropyl (meth)acrylate, acid phosphooxymethyl (meth)acrylate, acid phosphooxypolyoxyethylene glycol mono(meth)acrylate, and acid phosphooxypolyoxypropylene glycol mono(meth)acrylate. Among these, vinylphosphonic acid, acid phosphooxyethyl methacrylate (= 2-(methacryloyloxy)ethyl phosphate), acid phosphooxypolyoxyethylene glycol monomethacrylate, and acid phosphooxypolyoxypropylene glycol monomethacrylate are preferred.

[0059] The structural formulas of vinylphosphonic acid, acid phosphooxyethyl methacrylate (= 2-(methacryloyloxy)ethyl phosphate), acid phosphooxypolyoxyethylene glycol monomethacrylate, and acid phosphooxypolyoxypropylene glycol monomethacrylate are represented by the following formulas (A-1) to (A-4).

[0060]

[0061] These can be prepared by known methods or obtained commercially from reagent suppliers. For example, acid phosphooxyethyl methacrylate (= 2-(methacryloyloxy)ethyl phosphate) is available under the product names: Fosmer M (manufactured by Unichemical Co., Ltd.) and Light Ester P-1M (manufactured by Kyoeisha Chemical Co., Ltd.), acid phosphooxypolyoxyethylene glycol monomethacrylate is available under the product name: Fosmer PE (manufactured by Unichemical Co., Ltd.), and acid phosphooxypolyoxypropylene glycol monomethacrylate is available under the product names: Fosmer PP (manufactured by Unichemical Co., Ltd.) and PPM-5P (manufactured by Toho Chemical Industry Co., Ltd.).

[0062] Specific examples of monomers of formula (B) above include dimethylaminoethyl (meth)acrylate, diethylaminoethyl (meth)acrylate, dimethylaminopropyl (meth)acrylate, 2-(t-butylamino)ethyl (meth)acrylate, and methacloylcholinchloride. Among these, dimethylaminoethyl (meth)acrylate, methacloylcholinchloride, or 2-(t-butylamino)ethyl (meth)acrylate are preferred. These can be prepared by known methods or obtained commercially from reagent suppliers.

[0063] Specific examples of monomers of formula (C) above include 4-benzoylphenyl (meth)acrylate and 4-benzoyl-3-hydroxyphenyl (meth)acrylate. These can be prepared by known methods or obtained commercially from reagent suppliers.

[0064] The proportion (molar ratio) of the monomer represented by formula (A) to the total monomer components in the monomer mixture is, for example, 3 to 80 mol%, preferably 5 to 70 mol%, more preferably 10 to 70 mol%, even more preferably 10 to 60 mol%, and particularly preferably 20 to 50 mol%. There may be two or more monomers represented by formula (A). Similarly, the proportion (molar ratio) of the monomer represented by formula (B) to the total monomer components in the monomer mixture is, for example, 3 to 80 mol%, preferably 5 to 70 mol%, more preferably 10 to 60 mol%, even more preferably 10 to 50 mol%, and particularly preferably 20 to 50 mol%. There may be two or more monomers represented by formula (B). By setting the anionic monomer (A) and cationic monomer (B) to these ranges, the resulting copolymer can exhibit excellent ability to suppress the adhesion of biomolecules.

[0065] Furthermore, the total ratio of the anionic monomer represented by formula (A) and the cationic monomer represented by formula (B) to the total monomer components in the monomer mixture is 40 mol% or more, preferably in the range of 40 to 90 mol%, more preferably in the range of 50 to 80 mol%, and particularly preferably in the range of 50 to 70 mol%. There are no particular limitations on the ratio (molar ratio) of the anionic monomer represented by formula (A) to the cationic monomer represented by formula (B), but it is preferably in the range of 1:5 to 5:1, more preferably in the range of 1:2 to 2:1, and particularly preferably in the range of 1:1.5 to 1.5:1.

[0066] The proportion (molar ratio) of the monomer represented by formula (C) to the total monomer components in the monomer mixture is preferably less than 50 mol%, more preferably in the range of 1 to 50 mol%, even more preferably in the range of 5 to 40 mol%, and particularly preferably in the range of 5 to 30 mol%. There may be two or more monomers represented by formula (C). By setting the proportion of the monomer represented by formula (C) within this range, it is believed that the benzophenone skeleton portion contained in the monomer represented by formula (C) forms crosslinks (i.e., hardens) between other skeleton portions in the copolymer and the copolymer and the substrate, for example, by light irradiation or radiation irradiation, thereby improving adhesion to the substrate and solvent resistance.

[0067] The copolymer of the present invention is not particularly limited as long as it is obtained by polymerizing a monomer mixture containing a monomer represented by formula (A), a monomer represented by formula (B), and a monomer represented by formula (C), and the monomer mixture may contain other monomers other than the monomer represented by formula (A) / formula (B) / formula (C) as long as it does not impair the purpose of the present invention. Such a copolymer may contain a total of 40 mol% or more, preferably 50 mol% or more, and more preferably 60 mol% or more of the monomer represented by formula (A) / formula (B) / formula (C) with respect to the total monomer components in the monomer mixture.

[0068] In another embodiment, the copolymer according to the present invention further comprises monomers represented by formulas (A), (B), and (C), in addition to formula (D): (In the formula, Tｄ Q ｄ , R ｄ The significance and preferred embodiment of the above formula (d1) can be obtained by a manufacturing method that includes a step of polymerizing a monomer mixture containing a monomer represented by the above formula (d1).

[0069] Specific examples of monomers of formula (D) above include linear or branched alkyl esters of (meth)acrylic acid such as butyl (meth)acrylate, 2-ethylhexyl (meth)acrylate, lauryl (meth)acrylate, and stearyl (meth)acrylate; cyclic alkyl esters of (meth)acrylic acid such as cyclohexyl (meth)acrylate and isobornyl (meth)acrylate; aralkyl esters of (meth)acrylic acid such as benzyl (meth)acrylate and phenethyl (meth)acrylate; styrene monomers such as styrene, methylstyrene, and chloromethylstyrene; vinyl ether monomers such as methyl vinyl ether and butyl vinyl ether; and vinyl ester monomers such as vinyl acetate and vinyl propionate. Among these, linear or branched alkyl esters of (meth)acrylic acid such as butyl (meth)acrylate, 2-ethylhexyl (meth)acrylate, lauryl (meth)acrylate, and stearyl (meth)acrylate are preferably used. These can be prepared by known methods or obtained commercially from reagent suppliers.

[0070] The proportion (molar ratio) of the monomer represented by formula (D) to the total monomer components in the monomer mixture is preferably 0 to 50 mol%, more preferably 0 to 45 mol%, and particularly preferably 10 to 40 mol%. There may be two or more monomers represented by formula (D). By setting the monomer represented by formula (D) within this range, it is useful for maintaining the copolymer's excellent ability to suppress the adhesion of biomaterials and improving its adhesion to the substrate.

[0071] The copolymer of the present invention is not particularly limited as long as it is obtained by polymerizing a monomer mixture containing a monomer represented by formula (A), a monomer represented by formula (B), a monomer represented by formula (C), and a monomer represented by formula (D), and the monomer mixture may contain other monomers other than those represented by formula (A) / formula (B) / formula (C) / formula (D) as long as the objective of the present invention is not impaired. Such a copolymer may contain a total of 70 mol% or more, preferably 80 mol% or more, and more preferably 90 mol% or more of the monomers represented by formula (A) / formula (B) / formula (C) / formula (D) with respect to the total monomer components in the monomer mixture.

[0072] In yet another embodiment, the copolymer according to the present invention may be obtained by a manufacturing method that includes polymerizing a monomer mixture containing monomers represented by formulas (A), (B), and (C), and optionally a monomer represented by formula (D), in addition to any other component. Examples of such optional components include difunctional monomers having two or more carbon-carbon unsaturated bonds. Specifically, a difunctional monomer is a monomer having two or more carbon-carbon double bonds, and examples include polyfunctional (meth)acrylate compounds, polyfunctional (meth)acrylamide compounds, polyfunctional polyesters, or isoprene compounds. The polyfunctional monomer is preferably a difunctional monomer, and more preferably a difunctional (meth)acrylate compound or a difunctional (meth)acrylamide compound.

[0073] Examples of difunctional (meth)acrylate compounds are shown below: (E) and (F): (In the formula, T ｅ , T ｆ , U ｆ , R ｅ , R ｆ Examples of monomers represented by formulas (e1) and (f1) include those whose meanings and preferred embodiments are the same as those of n3 and n4.

[0074] Specific examples of the difunctional monomer of formula (E) above include ethylene glycol di(meth)acrylate, triethylene glycol di(meth)acrylate, and propylene glycol di(meth)acrylate. Specific examples of the difunctional monomer of formula (F) above include bis(methacryloyloxymethyl) phosphate, bis[(2-methacryloyloxy)ethyl] phosphate, bis[3-(methacryloyloxy)propyl] phosphate, or difunctional monomers derived from formulas (A-3) or (A-4) above. These can be prepared by known methods or obtained commercially from reagent suppliers. Alternatively, these compounds may be included as by-products in monomers (preparations or reagents) of formula (A).

[0075] The ratio (molar ratio) of the difunctional monomer to the total monomer components in the monomer mixture is preferably 0 to 40 mol%, more preferably 0 to 30 mol%, and particularly preferably 1 to 30 mol%. Furthermore, there may be two or more types of difunctional monomers. By setting the difunctional monomer within this range, gelation of the solid content during manufacturing due to excessive crosslinking can be suppressed, and manufacturing can be facilitated.

[0076] The copolymer of the present invention is obtained by polymerizing a monomer mixture comprising monomers represented by formulas (A), (B), and (C), and optionally a monomer represented by formula (D) and / or any other monomer. Polymerization can be carried out by methods known in themselves (for example, the methods described in Japanese Patent Application Publication No. 2014-162865 and International Publication No. 2020 / 040247). For example, it can be synthesized by methods such as radical polymerization, anionic polymerization, and cationic polymerization, which are common methods for synthesizing (meth)acrylic polymers. Various methods are possible, including solution polymerization, suspension polymerization, emulsion polymerization, and bulk polymerization.

[0077] Polymerization can be prepared, for example, by a manufacturing method that includes the step of reacting (polymerizing) monomers represented by formulas (A), (B), and (C) (and optionally, monomers represented by formula (D) and / or any other monomers) in a solvent.

[0078] The reaction conditions involve heating a reaction vessel containing various raw materials (monomers, solvents, initiators, etc.) in an oil bath or the like to 50°C to 200°C, and stirring for 1 to 48 hours, more preferably 80°C to 150°C, for 5 to 30 hours, to allow the polymerization reaction to proceed and obtain the copolymer according to the present invention. A nitrogen atmosphere is preferred for the reaction atmosphere.

[0079] The solvent used in the polymerization reaction may be water, phosphate buffer, alcohols such as ethanol, or a mixed solvent combining these, but it is preferable to include water or ethanol. Furthermore, it is preferable to include water or ethanol in an amount of 10 to 100% by mass. Furthermore, it is preferable to include water or ethanol in an amount of 50 to 100% by mass. Furthermore, it is preferable to include water or ethanol in an amount of 80 to 100% by mass. Furthermore, it is preferable to include water or ethanol in an amount of 90 to 100% by mass. Preferably, the total amount of water and ethanol is 100% by mass.

[0080] The reaction procedure may involve adding all the raw materials to a reaction solvent at room temperature and then heating to the above temperature to polymerize them, or adding all or part of the mixture of raw materials dropwise to a preheated solvent. For example, since the anionic monomer represented by formula (A) is a monomer that readily associates, it may be added dropwise to the reaction solvent in small amounts so that it can disperse quickly when added to the reaction system. In this case, the reaction solvent may be heated (e.g., 40°C to 100°C) to increase the solubility of the monomer and polymer.

[0081] To efficiently advance polymerization reactions, it is desirable to use polymerization initiators, particularly radical polymerization initiators. Examples of radical polymerization initiators include azo polymerization initiators such as dimethyl 1,1'-azobis(1-cyclohexanecarboxylate) (VE-073, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), 2,2'-azobis(2,4-dimethylvaleronitrile) (V-65, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), 2,2'-azobis(isobutyronitrile) (AIBN, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), 2,2'-azobis[N-(2-carboxyethyl)-2-methylpropionamidine]n hydrate (VA-057, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), and 2,2'-(N-butyl-2-methylpropionamide) (VAm-110, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.).

[0082] The amount of polymerization initiator added is 0.05% to 10% by mass, relative to the total mass of monomers used in polymerization.

[0083] After the reaction is complete, the obtained copolymer may be isolated and purified by known methods, such as adding a poor solvent to the reaction solution, or replacing the medium of the reaction solution with a suitable solvent and performing dialysis. Alternatively, the reaction solution may be used as is as a copolymer-containing solution to prepare the coating film-forming composition of the present invention.

[0084] <Composition> In one embodiment, the composition of the present invention comprises the copolymer and solvent described above. The solvent may be derived from the reaction solution of the copolymer or may be added separately.

[0085] Solvents included in the composition of the present invention include water, phosphate-buffered saline (PBS), and alcohols. Examples of alcohols include C2-C6 alcohols, such as ethanol, propanol, isopropanol, 1-butanol, 2-butanol, isobutanol, t-butanol, 1-pentanol, 2-pentanol, 3-pentanol, 1-heptanol, 2-heptanol, 2,2-dimethyl-1-propanol (= neopentyl alcohol), 2-methyl-1-propanol, 2-methyl-1-butanol, 2-methyl-2-butanol (= t-amyl alcohol), 3-methyl-1-butanol, 3-methyl-3-pentanol, cyclopentanol, 1-hexanol, 2-hexanol, 3-hexanol, 2,3- Examples include dimethyl-2-butanol, 3,3-dimethyl-1-butanol, 3,3-dimethyl-2-butanol, 2-ethyl-1-butanol, 2-methyl-1-pentanol, 2-methyl-2-pentanol, 2-methyl-3-pentanol, 3-methyl-1-pentanol, 3-methyl-2-pentanol, 3-methyl-3-pentanol, 4-methyl-1-pentanol, 4-methyl-2-pentanol, 4-methyl-3-pentanol, and cyclohexanol. These can be used individually or as mixed solvents of combinations thereof, but from the viewpoint of dissolving the copolymer, they are preferably selected from water, PBS, ethanol, and propanol.

[0086] The composition of the present invention can be provided as a coating film-forming composition. In particular, the composition of the present invention is preferably a coating film-forming composition that has the ability to suppress the adhesion of biological substances. When provided as a coating film-forming composition, there are no particular limitations on the concentration of solids as long as a uniform coating film can be formed, but 0.01 to 50% by mass is desirable.

[0087] Furthermore, in addition to the copolymer and solvent described above, the coating film-forming composition of the present invention may also contain other substances as needed, provided that they do not impair the performance of the resulting coating film. Examples of other substances include preservatives, surfactants, primers to improve adhesion to the substrate, antifungal agents, and sugars.

[0088] To adjust the ion balance of the copolymer in the coating film-forming composition according to the present invention, the process of obtaining the coating film of the present invention may further include a step of pre-adjusting the pH of the coating film-forming composition. pH adjustment may be carried out, for example, by adding a pH adjusting agent to a composition containing the copolymer and a solvent, and setting the pH of the composition to 1 to 13, preferably 1.5 to 8.5, more preferably 1.5 to 5.5, or preferably 8 to 13, even more preferably 10 to 13. The type and amount of pH adjusting agent that can be used are appropriately selected according to the concentration of the copolymer and the ratio of its anions to cations.

[0089] Examples of pH adjusters include organic amines such as ammonia, triethylamine, diethanolamine, pyridine, N-methyl-D-glucamine, and tris(hydroxymethyl)aminomethane; alkali metal hydroxides such as potassium hydroxide and sodium hydroxide; alkali metal halides such as potassium chloride and sodium chloride; inorganic acids such as sulfuric acid, phosphoric acid, hydrochloric acid, and carbonic acid, or their alkali metal salts; quaternary ammonium cations such as choline, or mixtures thereof (for example, buffer solutions such as phosphate-buffered saline).

[0090] <Coating Film / Cured Product> In one embodiment, the coating film of the present invention is a cured product of a coating film-forming composition. The cured product can be formed by applying the coating film-forming composition according to the present invention to at least a portion of the surface of a substrate and curing it. There are no particular restrictions on the application method, and conventional application methods such as spin coating, dip coating, spray coating, and solvent casting can be used.

[0091] Specific application methods include, for example, immersing a substrate as described later in the coating film-forming composition, adding the coating film-forming composition to a container and letting it stand for a predetermined time, or applying the coating film-forming composition to the surface of a container or plate. In the case of a container, such as a cell culture vessel, the coating film-forming composition is added to the container and left to stand for a predetermined time. Addition can be done, for example, by adding 0.5 to 1 times the total volume of the container's coating film-forming composition using a syringe or the like. The standing time is performed by appropriately selecting the time and temperature depending on the material of the substrate and the components of the coating film-forming composition, but for example, it is performed from 1 minute to 24 hours, preferably from 5 minutes to 3 hours, at 10 to 80°C. This allows an uncured coating film to form on at least a part, preferably over the entire surface, of the container.

[0092] Next, the coated film may be subjected to a drying process. The drying process can be carried out under air or vacuum at a temperature in the range of -200 to less than 200°C, for example, at room temperature (10°C to 35°C, for example, 25°C), but to dry more quickly, it may be dried at, for example, 40°C to 100°C. Alternatively, a drying process at extremely low to low temperatures (around -200°C to -30°C) using the freeze-drying method may be used. Freeze-drying is also called vacuum freeze-drying, and is a method in which the material to be dried is cooled with a refrigerant and the solvent is removed by sublimation under vacuum. Common refrigerants used in freeze-drying include a mixture of dry ice and methanol (-78°C), liquid nitrogen (-196°C), etc.

[0093] Before or after the drying step, the coated film may be washed with one or more solvents selected from aqueous solutions containing water and electrolytes (for example, running water washing or ultrasonic washing) in order to remove any remaining impurities, unreacted monomers, etc., or to adjust the ion balance of the copolymer. The aqueous solution containing water and electrolytes may be heated to a temperature range of, for example, 40°C to 95°C. The aqueous solution containing electrolytes is preferably PBS, physiological saline (containing only sodium chloride), Dulbecco phosphate-buffered physiological saline, Tris-buffered physiological saline, HEPES-buffered physiological saline, and Veronal-buffered physiological saline, with PBS being particularly preferred.

[0094] Next, the coated surface is subjected to a curing process to form a coating film containing the cured copolymer of the present invention. The curing process can be carried out by irradiating the coated surface with light or radiation.

[0095] Light irradiation is typically carried out by irradiating with ultraviolet light in a high-humidity environment (e.g., 50-90% RH) or in the ambient environment. Examples of ultraviolet light sources that can be used include UV-LEDs, low, medium, or high-pressure mercury lamps, mercury xenon lamps, metal halide lamps, tungsten lamps, arc lamps, excimer lamps, excimer lasers, semiconductor lasers, YAG lasers, and various ultraviolet irradiation devices. The ultraviolet irradiation dose (cumulative light dose) is, for example, approximately 10-5000 mJ / cm². ２ That is the case.

[0096] Radiation irradiation means, for example, irradiation with gamma rays, X-rays, or electron beams, preferably gamma rays or X-rays, more preferably gamma rays.

[0097] Furthermore, radiation irradiation is preferable because it allows for sterilization at the same time as hardening. Sterilization by radiation can be performed in the final packaged form, with the object to be sterilized (the substrate on which the coating film is formed) sealed in a container, and can be performed at room temperature. Therefore, it is a safe sterilization method that does not involve concerns about material changes or damage caused by high-temperature treatment, as in sterilization by heating methods (e.g., high-pressure steam method, dry heat method), nor does it involve concerns about harmful residues such as toxic gases, as in sterilization by gas methods (e.g., ethylene oxide gas method). In addition, radiation sterilization has many advantages, such as easy control of the sterilization process and the ability to continuously sterilize large quantities of products under the same conditions.

[0098] Radiation hardening can be carried out using doses similar to those used in conventional sterilization processes. For example, a gamma ray irradiation dose of about 5 to 40 kGy is sufficient, preferably 10 to 30 kGy.

[0099] The coating film containing the cured copolymer of the present invention obtained in this manner can be used as a coating film having the ability to inhibit the adhesion of biomolecules. Due to the crosslinking structures formed between functional groups in the copolymer upon curing (for example, between the carbonyl group of the benzophenone skeleton and the polymer main chain and / or hydrophobic groups), and the crosslinking structures formed between the copolymer and the substrate material, the cured product exhibits excellent adhesion (particularly resistance to solvents) and excellent ability to inhibit the adhesion of biomolecules. In this invention, "ability to inhibit the adhesion of biomolecules" means that the relative amount of adhesion (%) when comparing a substrate on which a coating film has been formed with a substrate on which a coating film has not been formed is 50% or less, preferably 30% or less, and more preferably 20% or less. For example, in a protein adhesion test performed by the method described in the example (Test Example 2) later, when comparing the average absorbance of a plate on which the coating film of the present invention is formed with the average absorbance of a plate on which the coating film is not formed, the relative adhesion amount (%) ((average absorbance of the plate on which the coating film is formed) / (average absorbance of the uncoated plate) × 100) is 50% or less, preferably 30% or less, and more preferably 20% or less.

[0100] The coating film of the present invention may be subjected to a further cleaning step after the curing step. The above cleaning may be carried out by known methods, but running water cleaning or ultrasonic cleaning is preferable. Examples of cleaning solvents include water, aqueous solutions containing electrolytes, and alcohols. Here, the aqueous solution containing electrolytes is preferably PBS, physiological saline (containing only sodium chloride), Dulbecco's phosphate-buffered physiological saline, Tris-buffered physiological saline, HEPES-buffered physiological saline, and Veronal-buffered physiological saline, with PBS being particularly preferred. The alcohol is preferably an alcohol having 2 to 6 carbon atoms, with ethanol being particularly preferred. As the cleaning solvent, an aqueous alcohol solvent, which is a mixture of water or an aqueous solution containing an electrolyte and an alcohol, is preferred, and an aqueous ethanol solvent, which is a mixture of water and ethanol, is more preferred. The cleaning solvent is usually used at room temperature (e.g., 10 to 35°C), but may also be heated to a range of, for example, 40 to 95°C. After adhesion, the coating film remains firmly attached to the substrate without dissolving even when washed with water, PBS, alcohol, etc., resulting in minimal change in film thickness before and after washing, i.e., minimal dissolution of the coating film into the solvent.

[0101] The film thickness of the coating film of the present invention is in the range of 1 to 1000 nm, preferably in the range of 5 to 500 nm, 10 to 300 nm, 10 to 200 nm, 10 to 100 nm, and 10 to 50 nm.

[0102] <Substrate> In this specification, the substrate may be a container, instrument, etc. having any structure. As described above, the coating film containing the cured copolymer of the present invention can be used as a coating film having the ability to suppress the adhesion of biological substances. Therefore, examples of such substrates include instruments for collecting or delivering biological substances (e.g., blood glucose meters, injection needles, catheters, etc.), containers for storing biological substances (e.g., bags, bottles, vials, ampoules, etc., specifically glass or plastic vials, blood bags, storage containers for antibody drugs, etc.), instruments for culturing, separating, isolating or analyzing biological substances (e.g., various filters for coarse filtration, microfiltration, ultrafiltration, tangental flow filtration (TFF), dialysis and reverse osmosis, carriers, cover slips and other microscope peripheral instruments, microfluidic devices including flow cytometers such as cell sorters, etc. Examples of materials include tubes, cell culture plates, cell spheroid arrays, cell separation columns, microchannel chips, microwell array chips, assay chips, biochips, magnetic beads, measuring cells for fully automated analyzers, etc.), biopharmaceutical manufacturing equipment and instruments (e.g., filters, reaction vessels, fluid delivery tubes (flow channels), transfer pipes, purification equipment, cell culture plates, etc.), prosthetic materials (e.g., implants, bone fixation materials, sutures, adhesion prevention membranes, artificial blood vessels, etc.), as well as drug delivery media such as vesicles, microparticles, and nanoparticles, materials for diagnostic equipment such as gastroscopes, and medical application materials such as microfibers, nanofibers, magnetic particles, and filters. By applying the coating film-forming composition to the surface of a substrate and curing it, a substrate with the ability to suppress the adhesion of biological substances can be manufactured. Here, "surface" refers to the surface that comes into contact with biological substances.

[0103] The cell culture vessel of the present invention is provided with a coating film of the present invention on at least a portion of its surface. Preferably, the coating film is formed over the entire surface on which cell culture takes place. Examples of cell culture vessels include dishes (Petri dishes) such as Petri dishes, tissue culture dishes, and multi-dishes commonly used for cell culture, flasks such as cell culture flasks and spinner flasks, bags such as plastic bags, Teflon® bags, and culture bags, plates such as microplates, microwell plates, multi-plates, and multi-well plates, chamber slides, tubes for cell culture or cryopreservation, trays, bottles such as roller bottles, culture vessels having internal stirring blades for stirring cell suspensions, large culture tanks, bioreactors, etc. Preferably, dishes, plates, and trays are used.

[0104] The fluid channel tube of the present invention has a coating film of the present invention on at least a portion of its surface. Preferably, the coating film is formed over the entire surface of the inner wall that comes into contact with biological material. Examples of fluid delivery tubes (fluid channel tubes) include medical tubes and bioprocess tubes. A fluid channel tube equipped with the coating film of the present invention can be manufactured by passing the coating film-forming composition of the present invention through the tube and then curing it. Therefore, the inner diameter of the fluid channel tube is not particularly limited as long as it is within the range through which the coating film-forming composition of the present invention can pass. Furthermore, the inner wall of the fluid channel tube can be easily cured by irradiation from the outside of the fluid channel tube. In the fluid delivery tube (fluid channel tube) of the present invention, various materials known as medical tubes can be used from the viewpoint of their mechanical strength, flexibility, chemical resistance, and biocompatibility. Specifically, polyolefins such as polyethylene (PE) and polypropylene (PP), polyvinyl chloride (PVC), ethylene-vinyl acetate copolymer (EVA), polyurethane (PU), silicone elastomers, or fluororesins such as polytetrafluoroethylene (PTFE), tetrafluoroethylene-hexafluoropropylene copolymer (FEP), and tetrafluoroethylene-perfluoroalkyl vinyl ether copolymer (PFA), as well as thermoplastic elastomers (TPE), can be used. However, the material is not limited to these, and other resin materials with equivalent properties may be used.

[0105] The filter of the present invention has a coating film of the present invention on at least a portion of its surface. Preferably, the coating film is formed over the entire surface that comes into contact with biological material. Examples of filters include medical filters and filters for biopharmaceutical manufacturing, such as depth filters, membrane filters, ultrafiltration filters, hollow fiber membrane filters, and dialysis / reverse osmosis membrane filters. For example, a dialysis filter equipped with the coating film of the present invention can be manufactured by passing the coating film-forming composition according to the present invention through a hollow fiber filter with a diameter of 0.1 to 500 μm, and then curing it. Furthermore, by performing curing by radiation, curing can be performed even in places where the irradiation light cannot reach (for example, inside the dialysis filter), and curing and sterilization can be performed simultaneously, thus shortening the process. Moreover, the filtration method as a form of use for the filter is not particularly limited, and various known methods can be appropriately adopted. Examples include, but are not limited to, cross-flow filtration, which filters by flowing the fluid parallel to the membrane surface; dead-end filtration, which captures solids by introducing the fluid perpendicular to the membrane surface; direct-flow filtration, which pressurizes the supply flow directly towards the membrane surface and allows it to permeate; and vacuum filtration, which extracts the filtrate by reducing the pressure.

[0106] Furthermore, the substrate material can be, for example, glass, metal, metal-containing compound or metalloid-containing compound, activated carbon, or resin. Examples of metals include typical metals: (alkali metals: Li, Na, K, Rb, Cs; alkaline earth metals: Ca, Sr, Ba, Ra); magnesium group elements: Be, Mg, Zn, Cd, Hg; aluminum group elements: Al, Ga, In; rare earth elements: Y, La, Ce, Pr, Nd, Sm, Eu; tin group elements: Ti, Zr, Sn, Hf, Pb, Th; iron group elements: Fe, Co, Ni; iron group elements: V, Nb, Ta; chromium group elements: Cr, Mo, W, U; manganese group elements: Mn, Re; precious metals: Cu, Ag, Au; platinum group elements: Ru, Rh, Pd, Os, Ir, Pt, etc. Examples of metal-containing compounds or metalloid-containing compounds include ceramics, which are sintered bodies whose basic component is a metal oxide and are hardened by heat treatment at high temperatures; semiconductors such as silicon; inorganic solid materials such as molded bodies of inorganic compounds such as metal oxides or metalloid oxides (silicon oxide, alumina, etc.), metal carbides or metalloid carbides, metal nitrides or metalloid nitrides (silicon nitride, etc.), metal borides or metalloid borides; and aluminum, nickel titanium, and stainless steel (SUS304, SUS316, SUS316L, etc.).

[0107] Since we expect the formation of a crosslinked structure between the copolymer and the substrate material of the present invention, the substrate material is preferably a resin. The resin may be a natural resin or its derivative, or a synthetic resin. Preferred natural resins or their derivatives include cellulose, cellulose triacetate (CTA), nitrocellulose (NC), cellulose immobilized with dextran sulfate, etc. Preferred synthetic resins include polyacrylonitrile (PAN), polyester polymer alloy (PEPA), polystyrene (PS), polysulfone (PSF), polyethylene terephthalate (PET), polymethyl methacrylate (PMMA), polyvinyl alcohol (PVA), polyurethane (PU), ethylene vinyl alcohol (EVAL), polyethylene (PE), polyester, polypropylene (PP), polyvinylidene fluoride (PVDF), polyethersulfone (PES), polycarbonate (PC), polyvinyl chloride (PVC), polytetrafluoroethylene (PTFE), ultra-high molecular weight polyethylene (UHPE), cycloolefin polymer (COP), cycloolefin copolymer (COC), polydimethylsiloxane (PDMS), acrylonitrile-butadiene-styrene resin (ABS), or Teflon®. For example, in the manufacturing of the cell culture vessel of the present invention, since high-temperature treatment is not required when coating the surface of the vessel with the coating film-forming composition so that it is present on at least a portion of the surface, resins with low heat resistance can also be used.

[0108] The substrate material may be one type or a combination of two or more types. Among these materials, it is preferable that the substrate be glass, silicon, silicon oxide, polystyrene (PS), polypropylene (PP), polyethersulfone (PES), polyethylene terephthalate (PET), polycarbonate (PC), polyvinyl chloride (PVC), Teflon (registered trademark), cycloolefin polymer (COP), cycloolefin copolymer (COC), polydimethylsiloxane (PDMS), or stainless steel (SUS304, SUS316, SUS316L, etc.) alone or a combination selected from these, and it is particularly preferable that the substrate be glass, polystyrene (PS), polypropylene (PP), stainless steel (SUS304, SUS316, SUS316L, etc.), cycloolefin polymer (COP), cycloolefin copolymer (COC), or polydimethylsiloxane (PDMS).

[0109] In the present invention, biomolecules include proteins, sugars, viruses, nucleic acids and cells or combinations thereof, or biological tissues and body fluids containing them. The proteins include fibrinogen, bovine serum albumin (BSA), human albumin, various globulins, β-lipoproteins, various antibodies (IgG, IgA, IgM), peroxidases, various complements, various lectins, fibronectin, lysozyme, von Willebrand factor (vWF), serum γ-globulin, pepsin, ovalbumin, insulin, histones, ribonucleases, collagen, and cytochrome c. The sugars include glucose, galactose, mannose, fructose, heparin, and hyaluronic acid. The nucleic acids include deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). The above cells include fibroblasts, bone marrow cells, B lymphocytes, T lymphocytes, neutrophils, erythrocytes, platelets, macrophages, monocytes, osteocytes, pericytes, dendritic cells, keratinocytes, adipocytes, mesenchymal cells, epithelial cells, epidermal cells, endothelial cells, vascular endothelial cells, hepatocytes, chondrocytes, cumulus cells, nervous system cells, glial cells, neurons, oligodendrocytes, microglia, astrocytes, cardiac cells, esophageal cells, muscle cells (e.g., smooth muscle cells or skeletal muscle cells), pancreatic beta cells, melanocytes, hematopoietic progenitor cells, mononuclear cells, embryonic stem cells (ES cells), embryonic tumor cells, embryonic germline stem cells, and artificial cells. Examples include pluripotent stem cells (iPS cells), neural stem cells, hematopoietic stem cells, mesenchymal stem cells, hepatic stem cells, pancreatic stem cells, muscle stem cells, germline stem cells, intestinal stem cells, cancer stem cells, hair follicle stem cells, and various cell lines (e.g., HCT116, Huh7, HEK293 (human fetal kidney cells), HeLa (human cervical cancer cell line), HepG2 (human hepatitis cell line), UT7 / TPO (human leukemia cell line), CHO (Chinese hamster ovary cell line), MDCK, MDBK, BHK, C-33A, HT-29, AE-1, 3D9, Ns0 / 1, Jurkat, NIH3T3, PC12, S2, Sf9, Sf21, High Five, Vero).

[0110] Furthermore, the biosubstances in the present invention may be substances that can be administered to and act upon a living organism. Examples of such substances include small molecule drugs such as peptides (cyclic peptides) and small molecule compounds, and biopharmaceuticals such as enzymes, blood coagulation and fibrinolytic factors, serum proteins, hormones, vaccines, interferons, erythropoietins, cytokines, toxins, antibodies, antibody-drug conjugates, and fusion proteins.

[0111] <Method for Measuring Molecular Weight> The weight-average molecular weight shown in the synthesis example below is the result obtained by Gel Filtration Chromatography (hereinafter abbreviated as GFC). (Measurement conditions) ・Apparatus: HLC-8320GPC (manufactured by Tosoh Corporation) ・GFC column: TSKgel GMPWXL ・Flow rate: 0.5 mL / min ・Eluent: Salt-containing water / organic mixed solvent ・Column temperature: 40°C ・Detector: RI ・Injection concentration: Polymer solids 0.1% by mass ・Injection volume: 100 μL ・Calibration curve: Cubic approximation curve ・Standard samples: Sodium polystyrene sulfonate (manufactured by Agilent) × 8 types

[0112] <Synthesis Example 1> 8.03 g of acid phosphooxypolypropylene glycol monomethacrylate (average number of moles of propylene oxide added: 5; molecular weight per phosphate group calculated by titration: 618) (product name: PPM-5P, manufactured by Toho Chemical Industries, Ltd.) was mixed with 3.36 g of approximately 80% aqueous solution of methacloylcholinchloride (manufactured by Tokyo Chemical Industries, Ltd.), 4.17 g of 2-ethylhexyl methacrylate (manufactured by Tokyo Chemical Industries, Ltd.), 1.38 g of 4-benzoylphenyl methacrylate (manufactured by Tokyo Chemical Industries, Ltd.), 64.8 g of ethanol (manufactured by Kanto Chemical Co., Ltd.), and 0.164 g of dimethyl-1,1'-azobis(1-cyclohexanecarboxylate) (product name: VE-073, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) and stirred uniformly to prepare a mixture. This mixture was added to a four-necked flask equipped with a condenser, and the flask was purged with nitrogen. The mixture was then heated and stirred until it reached the reflux temperature. This condition was maintained for 24 hours while heating and stirring. After the reaction was complete, the mixture was cooled to obtain a copolymer-containing solution with a solid content of approximately 20.4% by mass. The weight-average molecular weight of this polymer, as determined by GFC, was 48,000.

[0113] <Synthesis Example 2> Using 4.02 g of acid phosphooxypolypropylene glycol monomethacrylate (average number of moles of propylene oxide added: 5; molecular weight per phosphate group calculated by titration: 618) (product name: PPM-5P, manufactured by Toho Chemical Industry Co., Ltd.), 1.68 g of approximately 80% aqueous solution of methacloylcholinchloride (manufactured by Tokyo Chemical Industry Co., Ltd.), 1.55 g of 2-ethylhexyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 1.37 g of 4-benzoylphenyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 33.1 g of ethanol (manufactured by Kanto Chemical Co., Ltd.), and 0.083 g of dimethyl-1,1'-azobis(1-cyclohexanecarboxylate) (product name: VE-073, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), the copolymer-containing solution with a solid content of approximately 25.2% by mass was synthesized in the same manner as in Synthesis Example 1. The weight-average molecular weight of this polymer, according to GFC, was 45,000.

[0114] <Synthesis Example 3> To 1.71 g of acid phosphooxyethyl methacrylate (product name: Fosmer M, manufactured by Unichemical Co., Ltd.), 1.41 g of approximately 80% aqueous solution of methacloylcholinchloride (manufactured by Tokyo Chemical Industry Co., Ltd.), 1.75 g of butyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 0.37 g of 4-benzoylphenyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 6.93 g of ethanol (manufactured by Kanto Chemical Co., Ltd.), 4.54 g of pure water, and 0.148 g of 2,2'-azobis(N-(2-carboxyethyl)-2-methylpropionamidine) n-hydrate (product name: VA-057, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) were added and the mixture was stirred uniformly to prepare a mixed solution. Meanwhile, 10.44 g of ethanol (manufactured by Kanto Chemical Co., Ltd.) and 6.81 g of pure water were added to a four-necked flask equipped with a condenser, and the flask was purged with nitrogen. The mixture was then heated to the reflux temperature while stirring. While maintaining this condition, the above mixture was added dropwise, and the mixture was heated and stirred for 3 hours while maintaining the above conditions. After the reaction was complete, the mixture was cooled to obtain a copolymer-containing solution with a solid content of approximately 15.3% by mass. The weight-average molecular weight of this polymer, as determined by GFC, was 524,000.

[0115] <Synthesis Example 4> To 1.72 g of acid phosphooxyethyl methacrylate (product name: Fosmer M, manufactured by Unichemical Co., Ltd.), 1.43 g of approximately 80% aqueous solution of methacloylcholinchloride (manufactured by Tokyo Chemical Industry Co., Ltd.), 1.60 g of butyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 0.74 g of 4-benzoylphenyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 8.37 g of ethanol (manufactured by Kanto Chemical Co., Ltd.), 3.49 g of pure water, and 0.154 g of 2,2'-azobis(N-(2-carboxyethyl)-2-methylpropionamidine) n-hydrate (product name: VA-057, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) were added and the mixture was stirred uniformly to prepare a mixed solution. Meanwhile, 12.55 g of ethanol (manufactured by Kanto Chemical Co., Ltd.) and 5.22 g of pure water were added to a four-necked flask equipped with a condenser, and the flask was purged with nitrogen. The mixture was then heated to the reflux temperature while stirring. While maintaining this condition, the above mixture was added dropwise, and the mixture was heated and stirred for 3 hours while maintaining the above environment. After the reaction was complete, the mixture was cooled to obtain a copolymer-containing solution with a solid content of approximately 15.2% by mass. The weight-average molecular weight of this polymer, as determined by GFC, was 542,000.

[0116] <Synthesis Example 5> To 1.61 g of acid phosphooxyethyl methacrylate (product name: Fosmer M, manufactured by Unichemical Co., Ltd.), 1.44 g of approximately 80% aqueous solution of methacloylcholinchloride (manufactured by Tokyo Chemical Industry Co., Ltd.), 1.11 g of butyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 0.30 g of 4-benzoylphenyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 9.10 g of ethanol (manufactured by Kanto Chemical Co., Ltd.), 6.15 g of pure water, and 0.126 g of 2,2'-azobis(N-(2-carboxyethyl)-2-methylpropionamidine) n-hydrate (product name: VA-057, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) were added and the mixture was stirred uniformly to prepare a mixed solution. Meanwhile, 13.68 g of ethanol (manufactured by Kanto Chemical Co., Ltd.) and 9.25 g of pure water were added to a four-necked flask equipped with a condenser, and the flask was purged with nitrogen. The mixture was then heated to the reflux temperature while stirring. While maintaining this condition, the above mixture was added dropwise, and the mixture was heated and stirred for 3 hours while maintaining the above conditions. After the reaction was complete, the mixture was cooled to obtain a copolymer-containing solution with a solid content of approximately 10.7% by mass. The weight-average molecular weight of this polymer, as determined by GFC, was 23,000.

[0117] <Synthesis Example 6> To 1.81 g of acid phosphooxyethyl methacrylate (product name: Fosmer M, manufactured by Unichemical Co., Ltd.), 1.70 g of approximately 80% aqueous solution of methacloylcholinchloride (manufactured by Tokyo Chemical Industry Co., Ltd.), 0.78 g of butyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 0.29 g of 4-benzoylphenyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 9.51 g of ethanol (manufactured by Kanto Chemical Co., Ltd.), 6.30 g of pure water, and 0.127 g of 2,2'-azobis(N-(2-carboxyethyl)-2-methylpropionamidine) n-hydrate (product name: VA-057, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) were added and the mixture was stirred uniformly to prepare a mixed solution. Meanwhile, 13.86 g of ethanol (manufactured by Kanto Chemical Co., Ltd.) and 9.40 g of pure water were added to a four-necked flask equipped with a condenser, and the flask was purged with nitrogen. The mixture was then heated to the reflux temperature while stirring. While maintaining this condition, the above mixture was added dropwise, and the mixture was heated and stirred for 3 hours while maintaining the above conditions. After the reaction was complete, the mixture was cooled to obtain a copolymer-containing solution with a solid content of approximately 9.9% by mass. The weight-average molecular weight of this polymer, as determined by GFC, was 19,000.

[0118] <Synthesis Example 7> To 2.02 g of acid phosphooxyethyl methacrylate (product name: Fosmer M, manufactured by Unichemical Co., Ltd.), 1.96 g of approximately 80% aqueous solution of methacloylcholinchloride (manufactured by Tokyo Chemical Industry Co., Ltd.), 0.48 g of butyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 0.29 g of 4-benzoylphenyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 8.63 g of ethanol (manufactured by Kanto Chemical Co., Ltd.), 7.18 g of pure water, and 0.134 g of 2,2'-azobis(N-(2-carboxyethyl)-2-methylpropionamidine) n-hydrate (product name: VA-057, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) were added and the mixture was stirred uniformly to prepare a mixed solution. Meanwhile, 12.94 g of ethanol (manufactured by Kanto Chemical Co., Ltd.) and 10.77 g of pure water were added to a four-necked flask equipped with a condenser, and the flask was purged with nitrogen. The mixture was then heated to the reflux temperature while stirring. While maintaining this condition, the above mixture was added dropwise, and the mixture was heated and stirred for 3 hours while maintaining the above conditions. After the reaction was complete, the mixture was cooled to obtain a copolymer-containing solution with a solid content of approximately 10.2% by mass. The weight-average molecular weight of this polymer, as determined by GFC, was 107,000.

[0119] <Synthesis Example 8> 5.53 g of acid phosphooxypolypropylene glycol monomethacrylate (average number of moles of propylene oxide added: 5; molecular weight per phosphate group calculated by titration: 618) (product name: PPM-5P, manufactured by Toho Chemical Industry Co., Ltd.), 2.33 g of approximately 80% aqueous solution of methacloylcholinchloride (manufactured by Tokyo Chemical Industry Co., Ltd.), 0.53 g of 4-benzoylphenyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 31.82 g of ethanol (manufactured by Kanto Chemical Co., Ltd.), and 0.078 g of dimethyl-1,1'-azobis(1-cyclohexanecarboxylate) (product name: VE-073, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) were polymerized in the same manner as in Synthesis Example 1 to obtain a copolymer-containing solution with a solid content of approximately 20.9% by mass. The weight-average molecular weight of this polymer according to GFC was 67,000.

[0120] <Synthesis Example 9> 5.03 g of acid phosphooxypolypropylene glycol monomethacrylate (average number of moles of propylene oxide added: 5; molecular weight per phosphate group calculated by titration: 618) (product name: PPM-5P, manufactured by Toho Chemical Industry Co., Ltd.), 2.07 g of approximately 80% aqueous solution of methacloylcholinchloride (manufactured by Tokyo Chemical Industry Co., Ltd.), 1.08 g of 4-benzoylphenyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 31.35 g of ethanol (manufactured by Kanto Chemical Co., Ltd.), and 0.080 g of dimethyl-1,1'-azobis(1-cyclohexanecarboxylate) (product name: VE-073, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) were polymerized in the same manner as in Synthesis Example 1 to obtain a copolymer-containing solution with a solid content of approximately 31.4% by mass. The weight-average molecular weight of this polymer according to GFC was 68,000.

[0121] <Synthesis Example 10> 4.72 g of acid phosphooxypolypropylene glycol monomethacrylate (average number of moles of propylene oxide added: 5; molecular weight per phosphate group calculated by titration: 618) (product name: PPM-5P, manufactured by Toho Chemical Industries, Ltd.), 1.96 g of approximately 80% aqueous solution of methacloylcholinchloride (manufactured by Tokyo Chemical Industries, Ltd.), 1.74 g of 4-benzoylphenyl methacrylate (manufactured by Tokyo Chemical Industries, Ltd.), 32.40 g of ethanol (manufactured by Kanto Chemical Co., Ltd.), and 0.083 g of dimethyl-1,1'-azobis(1-cyclohexanecarboxylate) (product name: VE-073, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) were polymerized in the same manner as in Synthesis Example 1 to obtain a copolymer-containing solution with a solid content of approximately 23.4% by mass.

[0122] <Synthesis Example 11> To 35.01 g of acid phosphooxyethyl methacrylate (product name: Fosmer M, manufactured by Unichemical Co., Ltd.), 14.62 g of approximately 80% aqueous solution of methacloylcholinchloride (manufactured by Tokyo Chemical Industry Co., Ltd.), 3.35 g of 4-benzoylphenyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 67.43 g of ethanol (manufactured by Kanto Chemical Co., Ltd.), and 0.500 g of dimethyl-1,1'-azobis(1-cyclohexanecarboxylate) (product name: VE-073, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) were added and the mixture was stirred to prepare a mixed solution. Meanwhile, 101.25 g of ethanol (manufactured by Kanto Chemical Co., Ltd.) was added to a four-necked flask equipped with a condenser, and the flask was purged with nitrogen. The mixture was heated to the reflux temperature while stirring. While maintaining this condition, the above mixed solution was added dropwise, and the mixture was heated and stirred for 23 hours while maintaining the above environment after addition. After the reaction was complete, a copolymer-containing solution with a solid content of approximately 20.0% by mass was obtained by cooling. The weight-average molecular weight of this polymer, as determined by GFC, was 62,000.

[0123] <Comparative Synthesis Example 1> Using 8.03 g of acid phosphooxypolypropylene glycol monomethacrylate (average number of moles of propylene oxide added: 5; molecular weight per phosphate group calculated by titration: 618) (product name: PPM-5P, manufactured by Toho Chemical Industry Co., Ltd.), 3.36 g of approximately 80% aqueous solution of methacloylcholinchloride (manufactured by Tokyo Chemical Industry Co., Ltd.), 3.44 g of 2-ethylhexyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 56.4 g of ethanol (manufactured by Kanto Chemical Co., Ltd.), and 0.142 g of dimethyl-1,1'-azobis(1-cyclohexanecarboxylate) (product name: VE-073, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), the polymer was synthesized in the same manner as in Synthesis Example 1 to obtain a copolymer-containing solution with a solid content of approximately 19.9% ​​by mass. The weight-average molecular weight of this polymer according to GFC was 58,000.

[0124] <Comparative Synthesis Example 2> 4.02 g of acid phosphooxypolypropylene glycol monomethacrylate (average number of moles of propylene oxide added: 5; molecular weight per phosphate group calculated by titration: 618) (product name: PPM-5P, manufactured by Toho Chemical Industry Co., Ltd.) was mixed with 1.69 g of approximately 80% aqueous solution of methacloylcholinchloride (manufactured by Tokyo Chemical Industry Co., Ltd.), 0.64 g of 2-ethylhexyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 4.30 g of 4-benzoylphenyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 93.5 g of ethanol (manufactured by Kanto Chemical Co., Ltd.), and 0.103 g of dimethyl-1,1'-azobis(1-cyclohexanecarboxylate) (product name: VE-073, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) and stirred uniformly to prepare a mixture. This mixture was added to a four-necked flask equipped with a condenser, and the flask was purged with nitrogen. The mixture was then heated to the reflux temperature while stirring. This condition was maintained and heating and stirring continued, but a white precipitate formed within one hour.

[0125] <Comparative Synthesis Example 3> 4.03 g of acid phosphooxypolypropylene glycol monomethacrylate (average number of moles of propylene oxide added: 5; molecular weight per phosphate group calculated by titration: 618) (product name: PPM-5P, manufactured by Toho Chemical Industry Co., Ltd.) was mixed with 1.67 g of approximately 80% aqueous solution of methacloylcholinchloride (manufactured by Tokyo Chemical Industry Co., Ltd.), 3.45 g of 4-benzoylphenyl methacrylate (manufactured by Tokyo Chemical Industry Co., Ltd.), 79.8 g of ethanol (manufactured by Kanto Chemical Co., Ltd.), and 0.087 g of dimethyl-1,1'-azobis(1-cyclohexanecarboxylate) (product name: VE-073, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) and stirred uniformly to prepare a mixture. This mixture was added to a four-necked flask equipped with a condenser, the flask was purged with nitrogen, and the temperature was raised to the reflux temperature while stirring. This state was maintained and heating and stirring continued, but a white precipitate formed within 1 hour.

[0126] <Preparation Example 1> 9.00 g of the copolymer-containing solution obtained in Synthesis Example 1 was mixed with 37.7 g of ethanol and 44.9 g of pure water, and thoroughly stirred to prepare a coating film-forming composition. The pH was 2.4.

[0127] <Preparation Example 2> To 1.33 g of the copolymer-containing solution obtained in Synthesis Example 2, 7.29 g of ethanol and 8.28 g of pure water were added and thoroughly stirred to prepare a coating film-forming composition. The pH was 2.4.

[0128] <Preparation Example 3> 2.00 g of the copolymer-containing solution obtained in Synthesis Example 3 was mixed with 9.50 g of ethanol and 3.83 g of pure water, and thoroughly stirred to prepare a coating film-forming composition. The pH was 2.5.

[0129] <Preparation Example 4> To 2.00 g of the copolymer-containing solution obtained in Synthesis Example 4, 9.39 g of ethanol, 3.02 g of pure water, and 0.79 g of 1 mol / L hydrochloric acid (1N) (manufactured by Kanto Chemical Co., Ltd.) were added and thoroughly stirred to prepare a coating film-forming composition. The pH was 1.7.

[0130] <Preparation Example 5> 4.51 g of the copolymer-containing solution obtained in Synthesis Example 5 was mixed with 9.36 g of ethanol and 10.18 g of pure water, and thoroughly stirred to prepare a coating film-forming composition. The pH was 2.2.

[0131] <Preparation Example 6> 4.81 g of the copolymer-containing solution obtained in Synthesis Example 6 was mixed with 9.09 g of ethanol and 9.95 g of pure water and stirred thoroughly to prepare a coating film-forming composition. The pH was 2.2.

[0132] <Preparation Example 7> 4.51 g of the copolymer-containing solution obtained in Synthesis Example 7 was mixed with 9.02 g of ethanol and 9.45 g of pure water and stirred thoroughly to prepare a coating film-forming composition. The pH was 2.2.

[0133] <Preparation Example 8> 2.50 g of the copolymer-containing solution obtained in Synthesis Example 8 was mixed with 16.74 g of ethanol and 6.88 g of pure water, and thoroughly stirred to prepare a coating film-forming composition. The pH was 2.6.

[0134] <Preparation Example 9> 1.60 g of the copolymer-containing solution obtained in Synthesis Example 9 was mixed with 16.54 g of ethanol and 6.95 g of pure water, and thoroughly stirred to prepare a coating film-forming composition. The pH was 2.6.

[0135] <Preparation Example 10> 2.11 g of the copolymer-containing solution obtained in Synthesis Example 10 was mixed with 15.08 g of ethanol and 6.61 g of pure water, and thoroughly stirred to prepare a coating film-forming composition. The pH was 2.6.

[0136] <Preparation Example 11> To 3.00 g of the copolymer-containing solution obtained in Synthesis Example 6, 6.03 g of ethanol, 5.67 g of pure water, and 0.19 g of 1 mol / L hydrochloric acid (1N) (manufactured by Kanto Chemical Co., Ltd.) were added and thoroughly stirred to prepare a coating film-forming composition. The pH was 2.0.

[0137] <Preparation Example 12> To 1.50 g of the copolymer-containing solution obtained in Synthesis Example 11, 9.10 g of ethanol and 4.41 g of pure water were added and thoroughly stirred to prepare a coating film-forming composition. The pH was 2.6.

[0138] <Preparation Example 13> To 0.80 g of the copolymer-containing solution obtained in Synthesis Example 11, 10.45 g of ethanol and 4.75 g of pure water were added and thoroughly stirred to prepare a coating film-forming composition. The pH was 3.0.

[0139] <Comparative Preparation Example 1> 2.01 g of the copolymer-containing solution obtained in Comparative Synthesis Example 1 was mixed with 8.23 ​​g of ethanol and 9.86 g of pure water, and thoroughly stirred to prepare a coating film-forming composition. The pH was 2.4.

[0140] <Test Example 1: Evaluation of Coating Film Elution> The coating film-forming compositions obtained in Preparation Examples 1-10, 12, and 13, and Comparative Preparation Example 1, were each spin-coated onto HMDS-treated silicon wafers at 1500 rpm / 60 sec, and the solvent was dried to obtain a coating film (uncured). This coating film was exposed to UV light at 20 mW / cm using a high-pressure mercury lamp in an air atmosphere. ２The wafers were irradiated for 50 seconds. Subsequently, they were washed with a mixed solution of pure water and ethanol, then dried to obtain the coating films (cured products) of Examples 1 to 12 and Comparative Example 4 on HMDS-treated silicon wafers. The film thickness of each coating film (cured product) was measured using a spectroscopic ellipsometer, and this film thickness was defined as the initial film thickness. Next, each coating film (cured product) was immersed in PBS for 24 hours, then washed with pure water, dried, and the film thickness was measured again using a spectroscopic ellipsometer. The initial film thickness and the film thickness after PBS immersion were compared, and the residual film percentage after PBS immersion, with the initial film thickness set to 100%, was used as an indicator of elution. The results are shown in Table 1. Similarly, the coating film-forming compositions of Preparation Examples 1 and 2 were spin-coated onto HMDS-treated silicon wafers, the solvent was dried, and after washing with a mixed solution of pure water and ethanol and drying, the coating films (uncured products) were obtained. The coating film was placed in a plastic bag with an oxygen absorber (product name: Ageless®, manufactured by Mitsubishi Gas Chemical Co., Ltd.) and an oxygen indicator (product name: Ageless Eye®, manufactured by Mitsubishi Gas Chemical Co., Ltd.), sealed, and irradiated with 25 kGy of gamma rays to obtain the coating films (cured products) of Examples 13 and 14. Subsequently, the coating films (cured products) were subjected to the same elution test as described above, and the initial film thickness and the film thickness after PBS immersion were compared to calculate the residual film percentage. The results are shown in Table 1. Similarly, the coating film forming compositions of Preparation Examples 12 and 13 were spin-coated onto HMDS-treated silicon wafers, the solvent was dried, and the wafers were washed with a mixed solution of pure water and ethanol and dried to obtain the coating films (uncured products). These coating films were packaged and irradiated with 25 kGy of gamma rays to obtain the coating films (cured products) of Examples 15 and 16. Subsequently, the coating film (cured product) was subjected to the same dissolution test as described above, and the initial film thickness and the film thickness after PBS immersion were compared to calculate the residual film percentage. The results are shown in Table 1. Furthermore, similarly to the above, the coating film forming compositions of Preparation Examples 1, 2, and 12 were spin-coated onto HMDS-treated silicon wafers, the solvent was dried, and the film was washed with a mixed solution of pure water and ethanol and dried to obtain the coating films (uncured products) of Comparative Examples 1, 2, and 3. Subsequently, these coating films (uncured products) were subjected to the same dissolution test as described above, and the initial film thickness and the film thickness after PBS immersion were compared to calculate the residual film percentage.The results are shown in Table 1.

[0141]

[0142] <Test Example 2: Protein Adsorption Evaluation> (Preparation of Coated Plates) The coating film-forming compositions obtained in Preparation Examples 1 to 13 and Comparative Preparation Example 1 were each added to 5 wells of a 96-well plate (Corning, #3363, 0.32 mL volume, made of polypropylene (PP), and / or Corning, #9017, 0.32 mL volume, made of polystyrene (PS), and / or Corning, #2797, 0.25 mL volume, made of polyvinyl chloride (PVC)) at a rate of 150 μL / well. Only in Preparation Example 11, ethanol was added as a pretreatment at a rate of 150 μL / well, drained, washed, dried, and then irradiated with plasma for 1 minute using an oxygen plasma device (Izumi Kogyo Co., Ltd., IPSOLON, voltage 105V). After standing at room temperature for 1 hour, the liquid was drained and dried to obtain a coated (uncured) plate. This coating plate is exposed to UV light at 20 mW / cm using a high-pressure mercury lamp in an air atmosphere. ２The plates were irradiated for 50 seconds. Afterwards, each well was washed three times with a 200 μL mixed solution of pure water and ethanol, dried, and coated (cured) plates for Examples 17-30 and Comparative Example 8 were prepared. As a negative control, wells from uncoated 96-well plates (Corning, #3363, 0.32 mL volume, made of polypropylene, and / or Corning, #9017, 0.32 mL volume, made of polystyrene, and / or Corning, #2797, 0.25 mL volume, made of polyvinyl chloride (PVC)) were used. Similarly, the coating film-forming compositions of Preparation Examples 1 and 2 were added to the 96-well plates, drained, dried, washed with a mixed solution of pure water and ethanol, and dried to obtain coated (uncured) plates. The coated plate was placed in a plastic bag with an oxygen absorber (product name: Ageless®, manufactured by Mitsubishi Gas Chemical Co., Ltd.) and an oxygen indicator (product name: Ageless Eye®, manufactured by Mitsubishi Gas Chemical Co., Ltd.), sealed, and irradiated with 25 kGy of gamma rays to produce the coated (cured) plates of Examples 31 and 32. Similarly, the coating film-forming compositions of Preparation Examples 12 and 13 were added to a 96-well plate, drained, dried, washed with a mixed solution of pure water and ethanol, and dried to obtain the coated (uncured) plates. These coated plates were packaged and irradiated with 25 kGy of gamma rays to produce the coated (cured) plates of Examples 33 and 34. Furthermore, similarly, the coating film-forming compositions of Preparation Examples 1, 2 and 12 were added to a 96-well plate, drained, dried, washed with a mixed solution of pure water and ethanol, and dried to produce the coated (uncured) plates of Comparative Examples 5 to 7.

[0143] (Preparation of IgG-HRP Dilution Solution) Goat anti-mouse IgG antibody HRP conjugate (Southern Biotechnology Associates) was diluted with PBS to a concentration of 1 mg / g to prepare an IgG-HRP dilution solution.

[0144] (Protein adsorption evaluation) 100 μL / well of IgG-HRP diluent was added to each well of the plate prepared above, as well as to the negative control, and the plates were left to stand at room temperature for 30 minutes. After that, the IgG-HRP diluent was drained, and each well was washed three times with 200 μL of PBS. 100 μL / well of TMB solution (SureBlue, sera care) was added to each well, and after 1 minute, 100 μL / well of TMB STOP solution (sera care) was added to each well. As background, TMB solution and TMB STOP solution were similarly added to the wells to which IgG-HRP diluent had not been added. Absorbance at 450 nm and 650 nm was measured using a microplate reader (TECAN, infinite M200PRO). The absorbance at 450 nm was calculated by subtracting the background absorbance, and the average absorbance of 5 wells was obtained for each coating film-forming composition. The average absorbance of each coating film-forming composition was compared with that of the negative control, and the average absorbance of each coating film-forming composition, with the average absorbance of the negative control set to 100%, was defined as the protein adsorption rate. The results are shown in Table 2.

[0145]

[0146] <Test Example 3: Protein Adsorption Evaluation (Solvent Resistance)> Using the coating film-forming compositions obtained in Preparation Examples 1 to 7 and a 96-well plate (Corning, #3363, 0.32 mL volume, made of PP), 300 μL of ethanol was added to each well of the coated (cured) plates of Examples 35 to 41 and the coated (uncured) plates of Comparative Examples 9 and 10, which were prepared in the same manner as in Test Example 2. The plates were left to stand at room temperature for 24 hours. After that, each well was washed three times with 200 μL of pure water and dried to prepare coating plates for solvent resistance evaluation. Subsequently, protein adsorption evaluation was performed in the same manner as in Test Example 2. The results are shown in Table 3.

[0147]

[0148] In the case of uncured coating films that were not irradiated with UV or gamma rays, as shown in Comparative Examples 5 to 7 of Test Example 2, the protein adsorption inhibition ability was excellent, but as shown in Comparative Examples 1 to 3 of Test Example 1, the residual film rate was low and elution of the coating film was observed. Similarly, when a coating film-forming composition without benzophenone (Comparative Preparation Example 1) was used, as shown in Comparative Example 6 of Test Example 2, the protein adsorption inhibition ability was excellent, but as shown in Comparative Example 4 of Test Example 1, elution of the coating film was confirmed. In contrast to these, when a coating film-forming composition containing benzophenone was used and the coating film was irradiated with UV or gamma rays (cured), as shown in Examples 1 to 16 of Test Example 1, the residual film rate was high and elution of the coating film was suppressed, and as shown in Examples 17 to 34 of Test Example 2, the protein adsorption inhibition ability was excellent, confirming that it was a coating film with a balanced performance as desired. Furthermore, in Test Example 3, the coated (cured) plates of Examples 35 to 41 showed excellent protein adsorption inhibition ability even after immersion in ethanol, confirming that the coating film of the present invention exhibits high solvent resistance not only to aqueous solvents such as PBS but also to organic solvents such as ethanol. On the other hand, the coated (uncured) plates of Comparative Examples 9 and 10 showed a significant decrease in protein adsorption inhibition ability (a substantial increase in adsorption rate) after immersion in ethanol, suggesting that they had poor solvent resistance.

[0149] <Test Example 4: Evaluation of Protein Adsorption in Glass Vials> (Preparation of Coated Glass Vials) The coating film-forming composition obtained in Preparation Example 8 was added to glass vials (JG Finneran, #32009-1232) at a rate of 2 mL / vial. After standing at room temperature for 5 minutes, the liquid was drained and dried to obtain coated (uncured) glass vials. UV light at 20 mW / cm² was applied to these coated glass vials using a high-pressure mercury lamp in an air atmosphere. ２ The vials were irradiated for 50 seconds. Afterwards, each vial was washed with a 2.5 mL mixture of pure water and ethanol, dried, and the coated (cured) glass vials of Example 42 were prepared. An uncoated glass vial (JG Finneran, #32009-1232) was used as a negative control.

[0150] (Protein adsorption evaluation) 0.5 mL of IgG-HRP diluent was added to each of the coated glass vials and negative control prepared above, and the mixture was allowed to stand at room temperature for 30 minutes. After that, the IgG-HRP diluent was drained, and each vial was washed three times with 1 mL of PBS. 0.5 mL of TMB solution (SureBlue, sera care) was added to each vial, and after 1 minute, 0.5 mL of TMB STOP solution (sera care) was added to each vial and mixed. 200 μL of this mixture was dispensed into each of the 96-well transparent microplates (Corning, #9017). As background, 100 μL each of TMB solution and TMB STOP solution were added to the wells to which the above mixture had not been added. The absorbance at 450 nm was measured using Spectramax (manufactured by Molecular Devices Japan Co., Ltd.). The average absorbance of the three vials was calculated by subtracting the average background absorbance from the 450 nm absorbance obtained from the coated glass vials and the negative control. The average absorbance of the coated glass vials and the negative control in Example 42 was compared, and the average absorbance of the coated glass vials was calculated as the protein adsorption rate, with the average absorbance of the negative control set to 100%, resulting in a value of 2.1%. This confirmed that the coating film of the present invention exhibits excellent protein adsorption inhibition ability even on glass vials.

[0151] <Test Example 5: Evaluation of Protein Adsorption on a 0.22 μm Pore Size Membrane Filter> (Preparation of Coated Membrane Filter) A 0.22 μm pore size polyethersulfone (PES) membrane filter (GVS, #1214193) was set in a filter holder (Merck, #SX0002500). A 10 mL syringe filled with 1 mL of the coating film-forming composition obtained in Preparation Example 8 was connected to the filter holder and passed through the filter. The filter was removed from the filter holder and dried to obtain a coated (uncured) filter. UV light at 20 mW / cm² was applied to both sides of this coated filter using a high-pressure mercury lamp in an air atmosphere. ２The filter was irradiated for 50 seconds. After that, the coated filter was placed in the filter holder, and a 10 mL syringe filled with a mixed solution of pure water and ethanol was connected to the filter holder and passed through the filter. The filter was removed from the filter holder and dried to obtain the coated (cured) filter of Example 43. An uncoated PES membrane filter (GVS, #1214193) was used as a negative control.

[0152] (Preparation of BSA solution (2 mg / mL)) Bovine serum-derived albumin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd., #015-27053) was dissolved in PBS to a concentration of 2 mg / mL to prepare a BSA solution.

[0153] (Preparation of SDS aqueous solution (1 w / v%)) Sodium dodecyl sulfate (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd., #192-13981) was dissolved in ultrapure water to a concentration of 1 w / v% to prepare an SDS aqueous solution.

[0154] (Protein adsorption evaluation) The filter prepared above was immersed in 5 mL of PBS for more than 1 hour, and then immersed in 5 mL of BSA solution (2 mg / mL) for 2 hours to adsorb BSA. Next, the filter was washed by repeating the immersion in 5 mL of PBS three times. After that, the filter was immersed in 5 mL of SDS aqueous solution (1 w / v%) for 2 hours to dissolve the BSA adsorbed on the filter, and this was used as the SDS extraction sample.

[0155] (BCA Assay) A 62.5 μg / mL BSA solution was prepared by mixing 15 μL of BSA solution (2 mg / mL) with 465 μL of PBS. This solution was then diluted twice with PBS to prepare BSA solutions of 31.2, 15.6, 7.8, 3.9, and 1.95 μg / mL, which were used as a calibration curve BSA dilution series. The calibration curve BSA dilution series, SDS extract samples, and PBS and SDS aqueous solutions (1 w / v%) as background were added in 25 μL in four wells of a 96-well transparent microplate (Corning, #9017). A solution prepared by mixing solutions A and B of the Protein Assay BCA Kit (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd., #297-73101) in a 50:1 ratio was added to each well of the above-mentioned sample, and after gentle shaking, the mixture was heated at 60°C for 30 minutes. After cooling to room temperature, the absorbance at 562 nm was measured using Spectramax (manufactured by Molecular Devices Japan Co., Ltd.). The average absorbance of the four wells was calculated by subtracting the average background absorbance from the 562 nm absorbance obtained from the calibration BSA dilution series and the SDS extract sample. A calibration curve was created based on the average absorbance and BSA concentration of the calibration BSA dilution series, and the BSA concentration of the SDS extract sample was calculated. Furthermore, the amount of BSA adsorbed per unit area of ​​the negative control and coated filter was calculated based on the filter area. The results are shown in Table 4. We were able to confirm that the coating film of the present invention exhibits excellent protein adsorption suppression ability even on membrane filters.

[0156]

[0157] <Test Example 6: Evaluation of Filtration Performance of a 0.22 μm Pore Size Membrane Filter> (Measurement of Differential Pressure During Pure Water Flow) A coated (cured) filter, prepared in the same manner as in Example 43 using the coating film-forming composition obtained in Preparation Example 8, was set in a filter holder (Merck, #SX0002500). A 10 mL syringe filled with 10 mL of pure water was connected to the filter holder and the water was passed through the filter. Then, pure water was passed through the filter at a constant flow rate, and the differential pressure 1 minute after the start of flow was measured using a KrosFlo Research III TFF system (Spectrum Laboratories). As a negative control, the differential pressure was similarly measured for an uncoated PES membrane filter (GVS, #1214193), and as a comparative control, for a 0.22 μm pore size polyvinylidene fluoride (PVDF) membrane filter (Merck, #GVWP02500). The results are shown in Table 5. It was confirmed that the coating film of the present invention suppresses the increase in differential pressure of the membrane filter and can achieve both excellent protein adsorption inhibition and filtration performance.

[0158]

[0159] <Test Example 7: Evaluation of Protein Adsorption in Fluid Delivery Tubes (Flow Channel Tubes)> (Preparation of Coated Fluid Delivery Tubes) The coating film-forming composition obtained in Preparation Example 11 was filled into bioprocess tubes (Saint-Gobain, #374-125-2) and medical tubes (Saint-Gobain, #ADF00006) so that the inner walls of the tubes were fully immersed. After standing at room temperature for 5 minutes, the liquid was drained to obtain coated (uncured) fluid delivery tubes. These coated fluid delivery tubes were exposed to UV light at 20 mW / cm using a high-pressure mercury lamp in an air atmosphere. ２The tubes were irradiated for 50 seconds. Afterwards, each fluid delivery tube was washed with a mixed solution of pure water and ethanol in an amount 10 times that of the coating film-forming composition, dried, and coated (cured) bioprocess tubes of Example 44 and coated (cured) medical tubes of Example 45 were prepared. In addition, fluid delivery tubes that had been irradiated with plasma for 1 minute using an oxygen plasma device (IPSOLON, manufactured by Izumi Kogyo Co., Ltd., voltage 105V) after passing ethanol through them were subjected to the same coating procedure to prepare coated (cured) bioprocess tubes of Example 46 and coated (cured) medical tubes of Example 47. As a negative control, fluid delivery tubes without coating were used.

[0160] (Protein adsorption evaluation) The coated delivery tubes and negative control prepared above were cut to a length of 7 cm, and 0.5 mL of IgG-HRP diluent was added to each tube using a syringe. The tubes were then held at room temperature for 30 minutes. After that, the IgG-HRP diluent was drained, and each delivery tube was washed once with 30 mL of PBS. 0.5 mL of TMB solution (SureBlue, sera care) was held in the delivery tubes using a syringe, and after 1 minute, it was mixed with 0.5 mL of TMB STOP solution (sera care). 200 μL of this mixture was dispensed into 96-well transparent microplates (Corning, #9017). 100 μL each of TMB solution and TMB STOP solution were added as background. The absorbance at 450 nm was measured using a Nivo multimode microplate reader (manufactured by Leviti Japan Co., Ltd.). The absorbance at 450 nm obtained from the coated fluid delivery tube and the negative control was calculated by subtracting the average background absorbance. The absorbance of the coated fluid delivery tube in Example 44 was compared with that of the negative control, and the protein adsorption rate was calculated by setting the absorbance of the coated fluid delivery tube to 100%. The results are shown in Table 6. It was confirmed that the coating film of the present invention exhibits excellent protein adsorption inhibition ability even on the inner wall of the fluid delivery tube.

[0161]

[0162] Thus, the coating film containing the cured product of the present invention maintains the same excellent ability to suppress the adhesion of biomolecules as coating films obtained from conventional copolymers, while further improving adhesion to the substrate, particularly elution into the solvent after adhesion, resulting in a balanced performance of the desired characteristics. In particular, the coating film of the present invention is typically obtained by a manufacturing method that includes the steps of applying a composition containing the copolymer of the present invention to the surface of a substrate and curing the coated surface. However, since curing can be performed not only by light irradiation but also by radiation irradiation of the coated surface, curing can be performed even in places where irradiation light cannot reach (for example, inside multilayer filters or dialysis filters), and curing and sterilization can be performed simultaneously, thus shortening the process.

Claims

1. Formula (a1) below: (wherein T ａ , U ａ１ and U ａ２ each independently represent a hydrogen atom or a linear or branched alkyl group having 1 to 5 carbon atoms; Q ａ represents a single bond, an ester bond or an amide bond; R ａ represents a linear or branched alkylene group having 1 to 10 carbon atoms which may be substituted with a halogen atom; n1 represents an integer of 1 to 10), a repeating unit represented by formula (b1) below: (wherein T ｂ , U ｂ１ , U ｂ２ and U ｂ３ each independently represent a hydrogen atom or a linear or branched alkyl group having 1 to 5 carbon atoms; Q ｂ represents a single bond, an ester bond or an amide bond; R ｂ represents a linear or branched alkylene group having 1 to 10 carbon atoms which may be substituted with a halogen atom; An － represents an anion selected from the group consisting of halide ions, inorganic acid ions, hydroxide ions and isothiocyanate ions), a repeating unit represented by formula (c1) below: [wherein T ｃ represents a hydrogen atom or a linear or branched alkyl group having 1 to 5 carbon atoms; Q ｃ represents a single bond, an ester bond or an amide bond; R ｃ１ represents a single bond, a linear or branched alkylene group having 1 to 10 carbon atoms which may be substituted with a halogen atom, or a group of the formula: (wherein Alk is a linear or branched alkylene group having 1 to 5 carbon atoms which may be substituted with a hydroxy group, and n2 is an integer of 1 to 10); R ｃ２ and R ｃ３ represent a hydrogen atom, a hydroxy group, or a linear or branched alkoxy group having 1 to 5 carbon atoms], a copolymer comprising the repeating unit represented by the formula.

2. Furthermore, the following equation (d1): [In the formula, T ｄ Q represents a hydrogen atom or a linear or branched alkyl group having 1 to 5 carbon atoms; ｄ R represents a single bond, ether bond, or ester bond; ｄ The copolymer according to claim 1, comprising a repeating unit represented by [wherein the aryl portion is a linear or branched alkyl group having 1 to 18 carbon atoms, a cyclic hydrocarbon group having 3 to 10 carbon atoms, an aryl group having 6 to 10 carbon atoms, an aralkyl group having 7 to 15 carbon atoms, or an aryloxyalkyl group having 7 to 15 carbon atoms (wherein the aryl portion may be substituted with a linear or branched alkyl group having 1 to 5 carbon atoms, which may be substituted with a halogen atom)].

3. The copolymer according to claim 1, wherein the proportion of repeating units represented by formula (c1) contained in the copolymer is less than 50 mol%.

4. A composition comprising a copolymer according to any one of claims 1 to 3 and a solvent.

5. A coating film comprising a cured copolymer according to any one of claims 1 to 3.

6. The coating film according to claim 5, which has the ability to suppress the adhesion of biological substances.

7. A method for producing a coating film, comprising the steps of applying the composition described in claim 4 to the surface of a substrate, and curing the coated surface.

8. The manufacturing method according to claim 7, wherein curing is performed by irradiating the coated surface with light or radiation.

9. A container comprising the coating film described in claim 5.

10. The container according to claim 9, which is for cell culture or storage of biological materials.

11. A flow channel tube comprising the coating film described in claim 5.

12. A filter comprising the coating film described in claim 5.

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