Hydrophilic substrate and method for producing a hydrophilic substrate

Abstract

The present application provides a hydrophilic substrate having a hydrophilic polymer layer formed thereon with controlled surface uniformity and unevenness, and a method for manufacturing the same. A hydrophilic substrate having a hydrophilic polymer layer formed on the substrate surface represented by a group of -N(R 1 )2 and / or -N(R 2 )3 + (R 1 and R 2 are the same or different, and are a hydrogen atom or a substituted or unsubstituted hydrocarbon group which can contain a hetero atom).

Description

TECHNICAL FIELD

[0001] The present application relates to a hydrophilic substrate having a hydrophilic polymer layer formed on the surface of a substrate and a method for producing the same. BACKGROUND

[0002] In order to produce a device for capturing specific cells (blood cells, cancer cells present in blood and body fluid, etc.) in blood and body fluid, a technique of coating the surface of a substrate with a special polymer has been proposed.

[0003] However, among the special polymers, there are polymers that are difficult to coat to produce a smooth surface, and the unevenness of the surface can affect the capturing performance of specific cells, and thus, it is desirable to provide a substrate having a surface on which the capturing performance of specific cells such as cancer cells, etc. is excellent and the unevenness of which is controlled (see Patent Document 1, etc.).

[0004] PRIOR ART DOCUMENTS

[0005] PATENT DOCUMENTS

[0006] Patent Document 1: Japanese Patent Laid-Open No. 2005-523981 SUMMARY

[0007] PROBLEMS TO BE SOLVED BY THE INVENTION

[0008] The present application has been made to solve the above-described technical problems, and provides a hydrophilic substrate having a hydrophilic polymer layer on which the uniformity and the unevenness of the surface are controlled and a method for producing the same.

[0009] TECHNICAL SOLUTION

[0010] The present application relates to a hydrophilic substrate having a hydrophilic polymer layer formed on the surface of a substrate having a group represented by -N(R 1 )2and / or -N(R 2 )3 + , wherein R 1 and R 2 are each the same or different and are a hydrogen atom or a substituted or unsubstituted hydrocarbon group which can contain a hetero atom.

[0011] The above-described hydrophilic substrate is preferably a substrate into which a group represented by -N(R 1 )2and / or -N(R 2 )3 + is introduced by treating the surface of the substrate with a silane coupling agent having a group represented by -N(R 1 )2and / or -N(R 2 )3 + .

[0012] The hydrophilic substrate is preferably a layer of a hydrophilic polymer formed using a hydrophilic polymer solution and / or a hydrophilic polymer dispersion.

[0013] The hydrophilic substrate is preferably a layer of a hydrophilic polymer formed using a hydrophilic polymer solution and / or a hydrophilic polymer dispersion.

[0014] [Chemical Formula 1]

[0015]

[0016] (In the formula, R 51 represents a hydrogen atom or a methyl group, R 52 represents an alkyl group. p is 1 to 8, m is 1 to 5, and n represents the number of repetitions).

[0017] The hydrophilic substrate is preferably a layer of a hydrophilic polymer formed using a hydrophilic polymer solution and / or a hydrophilic polymer dispersion.

[0018] [Chemical Formula 2]

[0019]

[0020] (In the formula, R 51 , R 52 , p, and m are the same as described above).

[0021] The hydrophilic substrate is preferably a layer of a hydrophilic polymer formed using a hydrophilic polymer solution and / or a hydrophilic polymer dispersion.

[0022] The hydrophilic substrate is preferably a layer of a hydrophilic polymer formed using a hydrophilic polymer solution and / or a hydrophilic polymer dispersion.

[0023] In addition, the present application relates to a method for producing a hydrophilic substrate, in which a layer of a hydrophilic polymer is formed on a substrate surface having a group represented by -N(R 1 )2 and / or -N(R 2 )3 + (R 1 and R 2 are each the same or different and are a hydrogen atom or a substituted or unsubstituted hydrocarbon group that can include a hetero atom).

[0024] The method for producing a hydrophilic substrate is preferably a method in which the group represented by -N(R 1 )2 and / or -N(R 2 )3 + is formed on a substrate surface by using a substrate having a group represented by -N(R 1 )2 and / or -N(R 2 )3 +The silane coupling agent of the indicated group is introduced by treating the surface of the substrate.

[0025] The hydrophilic substrate production method preferably forms the hydrophilic polymer layer using a hydrophilic polymer solution and / or a hydrophilic polymer dispersion.

[0026] The hydrophilic substrate production method preferably forms the hydrophilic polymer layer using at least one hydrophilic polymer selected from the group consisting of polymers represented by the following formula (I).

[0027] [Chemical Formula 3]

[0028]

[0029] (In the formula, R 51 represents a hydrogen atom or a methyl group, R 52 represents an alkyl group. p is 1 to 8, m is 1 to 5, and n represents the number of repetitions).

[0030] The hydrophilic substrate production method preferably forms the hydrophilic polymer layer using a copolymer of at least one hydrophilic monomer selected from the group consisting of compounds represented by the following formula (II) and other monomers.

[0031] [Chemical Formula 4]

[0032]

[0033] (In the formula, R 51 , R 52 , p, and m are the same as described above).

[0034] The hydrophilic substrate production method preferably has a thickness of 10 to 1000 nm.

[0035] The hydrophilic substrate production method preferably has fibronectin adsorbed on the surface of the hydrophilic polymer layer.

[0036] Effects of the Invention

[0037] According to the present application, since a hydrophilic substrate having a hydrophilic polymer layer formed on the surface of a substrate having a group represented by -N(R 1 )2 and / or -N(R 2 )3 + (R 1 and R 2 are each independently a hydrogen atom or a substituted or unsubstituted hydrocarbon group that can include a hetero atom), a hydrophilic substrate having a hydrophilic polymer layer with controlled uniformity and unevenness of the surface can be provided. Therefore, it is expected that the capturing performance of specific cells such as cancer cells will be improved, and the unevenness of the capturing performance will be suppressed. BRIEF DESCRIPTION OF DRAWINGS

[0038] Figure 1 is an example of a photograph of the hydrophilic substrate produced in Examples 4, 5, and Comparative Example 1. DETAILED DESCRIPTION

[0039] The hydrophilic substrate of the present application is a substrate in which a hydrophilic polymer layer is formed on the surface of a substrate having a group represented by -N(R 1 )2and / or -N(R 2 )3 + (R 1 and R 2 are each the same or different and are a hydrogen atom or a substituted or unsubstituted hydrocarbon group which can contain a hetero atom). By forming a hydrophilic polymer layer on the surface of a substrate in which a group represented by -N(R 1 )2and / or -N(R 2 )3 + is introduced beforehand, a hydrophilic substrate in which the uniformity of the surface, the unevenness is controlled, the surface roughness is small, and the smoothness of the hydrophilic polymer layer coated on the designated site is high can be provided. Therefore, it is expected that the capturing performance of specific cells such as cancer cells is improved, and the unevenness of the capturing performance is suppressed.

[0040] The number of tumor cells (cancer cells, etc.) appearing in a body fluid such as circulating tumor cells in blood (several to several hundred per 1 mL of blood) is very small, and it is important to capture as many tumor cells present in the collected body fluid as possible for examination. In the present application, by coating a hydrophilic polymer on the surface of a substrate in which a group represented by -N(R 1 )2and / or -N(R 2 )3 + is introduced, a hydrophilic polymer layer in which the uniformity of the surface, the unevenness is controlled, the surface roughness is small, and the smoothness is high is formed. Moreover, the uniformity of the hydrophilic polymer layer, the unevenness has an influence on the capturing performance of specific cells such as cancer cells, and it is expected that by controlling the uniformity of the surface, the unevenness, the smoothness is improved, and thus an excellent specific cell capturing performance is exerted. In addition, it is expected that a capturing performance with less unevenness is exerted. Therefore, by measuring the number of tumor cells captured by the hydrophilic polymer layer on the surface of the substrate after the introduction of -N(R 1 )2and / or -N(R 2 )3 + groups, the number of tumor cells in the body fluid can be determined, and it is expected that the confirmation of the effect of cancer treatment, etc. is performed. In addition, the captured tumor cells are cultured, the effect of an anticancer agent, etc. is confirmed by the cells of the culture, and before the administration of the anticancer agent, etc., the effect of the anticancer agent, etc. can be confirmed in vitro, and at the same time, the selection of the anticancer agent, etc. also contributes.

[0041] Regarding the "substrate having a group represented by -N(R 1)2and / or -N(R 2 )3 + )2and / or -N(R 1 )2represents a group in which R 1 , -N(R 2 )3 + )3represents a group in which R 2 are the same or different, and are (1) a hydrogen atom or (2) a substituted or unsubstituted hydrocarbon group which can contain a hetero atom.

[0042] R 1 ~ R 2 The substituted or unsubstituted hydrocarbon group which can contain a hetero atom can be any of a linear, branched or cyclic group. As the hetero atom, there is no particular limitation, and examples include oxygen, nitrogen and the like. As the substituent, there is no particular limitation, and examples include a hydroxyl group, a halogen group (-CI, -Br and the like) and the like. These hetero atoms and substituents can be one or a plurality of.

[0043] The number of carbon atoms of the substituted or unsubstituted hydrocarbon group which can contain a hetero atom is preferably 1 to 20, more preferably 1 to 10, and further preferably 1 to 8.

[0044] As the substituted or unsubstituted hydrocarbon group which can contain a hetero atom, examples include a monovalent hydrocarbon group, a divalent hydrocarbon group and the like.

[0045] As the substituted or unsubstituted monovalent hydrocarbon group which can contain a hetero atom, examples include a substituted or unsubstituted linear alkyl group, a branched alkyl group, a cyclic alkyl group, a cyclic ether group, an aryl group, an aralkyl group, an alkoxy group and the like which can contain a hetero atom.

[0046] As the substituted or unsubstituted straight-chain alkyl group, branched alkyl group which can contain a hetero atom, there are exemplified methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, t-butyl group, pentyl group, hexyl group, heptyl group, 2-ethylhexyl group, octyl group, nonyl group, decyl group, and the like, and groups containing a hetero atom thereof. As the substituted or unsubstituted cyclic alkyl group which can contain a hetero atom, there are exemplified cyclopropyl group, cyclobutyl group, cyclopentyl group, cyclohexyl group, cycloheptyl group, cyclooctyl group, adamantyl group, 1-ethylcyclopentyl group, 1-ethylcyclohexyl group, and the like, and groups containing a hetero atom thereof. As the substituted or unsubstituted cyclic ether group which can contain a hetero atom, there are exemplified oxiranyl group, oxetanyl group, oxolanyl group, oxanyl group, oxepanyl group, oxocanyl group, oxonanyl group, oxecanyl group, oxete group, oxole group, dioxolanyl group, dioxiranyl group, dioxepanyl group, dioxecanyl group, and the like, and groups containing a hetero atom thereof. As the substituted or unsubstituted aryl group which can contain a hetero atom, there are exemplified phenyl group, tolyl group, xylyl group, biphenyl group, naphthyl group, anthryl group, phenanthryl group, and the like, and groups containing a hetero atom thereof. As the substituted or unsubstituted aralkyl group which can contain a hetero atom, there are exemplified benzyl group, phenethyl group, and the like, and groups containing a hetero atom thereof. As the substituted or unsubstituted alkoxy group which can contain a hetero atom, there are exemplified methoxy group, ethoxy group, n-propoxy group, isopropoxy group, n-butoxy group, sec-butoxy group, isobutoxy group, t-butoxy group, n-pentoxy group, n-hexyloxy group, and the like, and groups containing a hetero atom thereof.

[0047] As the substituted or unsubstituted divalent hydrocarbon group which can contain a hetero atom, there are exemplified substituted or unsubstituted alkylene group, alkenylene group, cycloalkylene group, cycloalkylalkylene group, arylene group, aralkylene group, oxyalkylene group, and the like, which can contain a hetero atom.

[0048] As specific examples of the substituted or unsubstituted alkylene group which can contain a hetero atom, there can be mentioned methylene, ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene, octylene, nonylene, decylene, 1,2- propylene, and groups containing a hetero atom thereof, and the like. As the substituted or unsubstituted alkenylene group which can contain a hetero atom, there can be mentioned ethenylene, 1- propenylene, 2-propenylene, and groups containing a hetero atom thereof, and the like. As the substituted or unsubstituted cycloalkylene group which can contain a hetero atom, there can be mentioned cyclohexylene and groups containing a hetero atom thereof, and the like. As the substituted or unsubstituted cycloalkylalkylene group which can contain a hetero atom, there can be mentioned cyclohexylmethylene and groups containing a hetero atom thereof, and the like. As the substituted or unsubstituted arylene group which can contain a hetero atom, there can be mentioned phenylene, benzylene, xylylene, and groups containing a hetero atom thereof, and the like. As the substituted or unsubstituted aroyl group which can contain a hetero atom, there can be mentioned benzylidene and groups containing a hetero atom thereof, and the like. As the substituted or unsubstituted oxyalkylene group which can contain a hetero atom, there can be mentioned oxyethylene, oxypropylene, oxybutylene, oxytetramethylene, and groups containing a hetero atom thereof, and the like.

[0049] As to the "substrate surface having a group represented by -N(R 1 )2and / or -N(R 2 )3 + As the substrate, there can be mentioned soda lime glass, borosilicate glass, and the like glass; polymethyl acrylate, polymethyl methacrylate, polyacrylic acid, polymethacrylic acid, and the like acrylic resin (polyacrylic resin); cycloolefin resin (polycycloolefin), carbonate resin (polycarbonate), styrene resin (polystyrene), polyester resin such as polyethylene terephthalate (PET); polydimethylsiloxane, and the like. Among them, in the coating of the hydrophilic polymer, it is preferable that the material constituting the substrate has high hydrophilicity, and it is preferable that the material is a soda lime glass or a polyacrylic resin.

[0050] As to the "substrate surface having a group represented by -N(R 1 )2and / or -N(R 2 )3 + The "substrate surface having a group represented by -N(R 1 )2and / or -N(R 2 )3 + The "substrate surface having a group represented by -N(R 1 )2and / or -N(R 2 )3 +The substrate surface is treated with a silane coupling agent containing the indicated group (silane coupling agent treatment); the substrate surface is treated with NH3 gas (NH3 gas treatment), etc. Among these, silane coupling agent treatment is preferred from the viewpoint of controlling the uniformity and unevenness of the hydrophilic polymer layer.

[0051] As a silane coupling agent, it has -N(R) 1 )2 and / or -N(R 2 )3 + The silane coupling agent with the indicated group is not particularly limited; for example, one with -N(R) can be used. 1 )2 and / or -N(R 2 )3 + The shown groups and -Si(R) 3 Compounds with the group shown in Figure 3 (R) 1 and R 2 They may be the same or different, consisting of hydrogen atoms or substituted or unsubstituted hydrocarbon groups that may contain heteroatoms; R 3 (Similar or different, including substituted or unsubstituted hydrocarbon groups that may contain heteroatoms, etc.)

[0052] "-Si(R 3 The R in ")3 represents a group" 3 Whether the hydrocarbon group is the same or different, it can be a substituted or unsubstituted hydrocarbon group that may contain heteroatoms. Examples of substituted or unsubstituted hydrocarbon groups that may contain heteroatoms include: [examples of R mentioned above]. 1 and R 2 It may contain a group identical to a heteroatom-substituted or unsubstituted monovalent hydrocarbon group. Preferably, it is a straight-chain alkyl, branched alkyl, cyclic alkyl, or alkoxy group. Specific examples of straight-chain alkyl, branched alkyl, cyclic alkyl, and alkoxy groups are listed above. 1 and R 2 The same functional groups (methyl, ethyl, methoxy, ethoxy, etc.). Additionally, R is preferred. 3 At least one of them is an alkoxy group, more preferably two, and even more preferably three.

[0053] As having -N(R) 1 )2 and / or -N(R 2 )3 + Groups and -Si(R) 3 Compounds with the group shown in )3, for example, compounds represented by the following formulas (1-1) and (1-2) may be preferred.

[0054] [Chemical Formula 5]

[0055]

[0056] (where R is in the formula) 11~R 12 the same or different, is a hydrogen atom, or a substituted or unsubstituted monovalent hydrocarbon group which can contain a hetero atom; R 16 ~R 18 the same or different, is a substituted or unsubstituted monovalent hydrocarbon group which can contain a hetero atom; R 19 is a substituted or unsubstituted divalent hydrocarbon group which can contain a hetero atom).

[0057] [Chemical Formula 6]

[0058]

[0059] (In the formula, R 11 ~R 13 the same or different, is a hydrogen atom, or a substituted or unsubstituted monovalent hydrocarbon group which can contain a hetero atom; R 16 ~R 18 the same or different, is a substituted or unsubstituted monovalent hydrocarbon group which can contain a hetero atom; R 19 is a substituted or unsubstituted divalent hydrocarbon group which can contain a hetero atom; X is a halogen).

[0060] as the "substituted or unsubstituted monovalent hydrocarbon group which can contain a hetero atom" of R 11 , R 12 , R 13 of formula (1-1), (1-2), for example, the same group as the aforementioned R 1 ~R 2 substituted or unsubstituted monovalent hydrocarbon group which can contain a hetero atom. Among them, R 11 , R 12 , R 13 are preferably a hydrogen atom, a linear or branched alkyl group which can contain a hetero atom, more preferably a hydrogen atom, a linear alkyl group, a branched alkyl group, -C p H 2p -O-C q H 2q+1 a group represented by (p, q are integers).

[0061] as the "substituted or unsubstituted monovalent hydrocarbon group which can contain a hetero atom" of R 16 , R 17 , R 18 of formula (1-1), (1-2), for example, the same group as the aforementioned R 1 ~R 2 substituted or unsubstituted monovalent hydrocarbon group which can contain a hetero atom. Among them, R 16 , R 17 , R 18 are preferably a substituted or unsubstituted alkoxy group which can contain a hetero atom, more preferably an alkoxy group, further preferably a methoxy group, an ethoxy group.

[0062] R 19 "substituted or unsubstituted divalent hydrocarbon group which can contain a hetero atom" of formula (1-1), (1-2) R 1 ~R 2 "substituted or unsubstituted divalent hydrocarbon group which can contain a hetero atom" of formula (1-1), (1-2) R 1 ~R 2 "substituted or unsubstituted divalent hydrocarbon group which can contain a hetero atom" of formula (1-1), (1-2) R 111 )-(R 112 )-indicates a group (R 111 indicates a hydrogen atom or an alkyl group; R 112 is -C p H 2p -(p is an integer of 1 to 8) indicates a group). Among them, R 19 is a substituted or unsubstituted alkylene group which can contain a hetero atom, preferably a group represented by -CO-N(R 111 )-(R 112 )-, more preferably a methylene group, an ethylene group, a trimethylene group, a tetramethylene group, a group represented by -CO-N(R 111 )-(R 112 )-.

[0063] As the halogen of formula (1-2) X, chlorine (chloride ion), bromine (bromide ion), etc. can be mentioned.

[0064] As the compound having a group represented by -N(R 21 )2and / or -N(R 23 )3 24 and a group represented by -Si(R 26 )3, a compound represented by the following formula (1-3) can also be preferably used.

[0065] [Chemical Formula 7]

[0066]

[0067] (in the formula, R 27 ~R 30 are the same or different, and are a hydrogen atom, or a substituted or unsubstituted monovalent hydrocarbon group which can contain a hetero atom; R 21 ~R 22 are the same or different, and are a substituted or unsubstituted monovalent hydrocarbon group which can contain a hetero atom; R 23 ~R 1 are a substituted or unsubstituted divalent hydrocarbon group which can contain a hetero atom; X is a halogen; and n indicates the number of repetitions).

[0068] R as in equation (1-3) 21 R 22 R 23 The phrase "may contain substituted or unsubstituted monovalent hydrocarbon groups with heteroatoms" can be exemplified by, for example, the aforementioned R 1 ~R 2 It can contain a group that is identical to a heteroatom-substituted or unsubstituted monovalent hydrocarbon group. Wherein, R 21 R 22 R 23 Preferably, it contains hydrogen atoms, or may contain straight-chain alkyl or branched alkyl groups containing heteroatoms, and more preferably hydrogen atoms.

[0069] R as in equation (1-3) 24 R 25 R 26 The phrase "may contain substituted or unsubstituted monovalent hydrocarbon groups with heteroatoms" can be exemplified by, for example, the aforementioned R 1 ~R 2 It can contain a group that is identical to a heteroatom-substituted or unsubstituted monovalent hydrocarbon group. Wherein, R 24 R 25 R 26 Preferably, it may contain substituted or unsubstituted alkoxy groups with heteroatoms, more preferably alkoxy groups, and even more preferably methoxy or ethoxy groups.

[0070] R as in equation (1-3) 27 R 28 R 29 R 30 The phrase "may contain substituted or unsubstituted divalent hydrocarbon groups with heteroatoms" can be exemplified by, for example, the aforementioned R 1 ~R 2 It can contain a group that is identical to a heteroatom-substituted or unsubstituted divalent hydrocarbon group. Wherein, R 27 R 28 R 29 R 30 Preferably, it may contain substituted or unsubstituted alkylene groups that contain heteroatoms, more preferably methylene, ethylene, trimethylene, or tetramethylene.

[0071] Examples of halogens for X in formula (1-3) include chlorine (chloride ion) and bromine (bromine ion).

[0072] The degree of polymerization of the structural unit represented by equation (1-3) is not particularly limited. It can be appropriately selected by considering the processability (coating properties) of the compound and its adhesion to the hydrophilic polymer layer.

[0073] The number-average molecular weight (Mn) of the compound (polymer) represented by the above formula (1-3) is preferably 500 to 6000, more preferably 1000 to 4000, and even more preferably 1000 to 3000.

[0074] As having -N(R) 1 Specific examples of silane coupling agents with the groups shown in )2 include: 3-aminopropyltrimethoxysilane, 3-aminopropyltriethoxysilane, 3-aminopropyltriisopropoxysilane, 3-aminopropylmethyldimethoxysilane, 3-aminopropylmethyldiethoxysilane, 3-(2-aminoethyl)aminopropyltrimethoxysilane, 3-(2-aminoethyl)aminopropylmethyldimethoxysilane, 3-(2-aminoethyl)aminopropyltriethoxysilane, 3-(2-aminoethyl)aminopropylmethyldiethoxysilane, 3-(2-aminoethyl)aminopropyltriisopropoxysilane, 3 Silanes containing amino groups include: -(2-(2-aminoethyl)aminoethyl)aminopropyltrimethoxysilane, 3-(6-aminohexyl)aminopropyltrimethoxysilane, 3-(N-ethylamino)-2-methylpropyltrimethoxysilane, N-phenyl-3-aminopropyltrimethoxysilane, N-benzyl-3-aminopropyltrimethoxysilane, N-vinylbenzyl-3-aminopropyltriethoxysilane, N-cyclohexylaminomethyltriethoxysilane, N-cyclohexylaminomethyldiethoxymethylsilane, N-phenylaminomethyltrimethoxysilane, and (2-aminoethyl)aminomethyltrimethoxysilane. Coupling agents; 3-Ureapropyltrimethoxysilane (1-[3-(trimethoxysilyl)propyl]urea), 3-Ureapropyltriethoxysilane, 3-Ureapropyltriisopropoxysilane, 3-Ureapropylmethyldimethoxysilane, 3-Ureapropylmethyldiethoxysilane, ureamethyltrimethoxysilane, ureamethyltriethoxysilane, ureamethyltriisopropoxysilane, ureamethylmethyldimethoxysilane, ureamethylmethyldiethoxysilane, 3-[(2-ureaethyl)amino]propyltrimethoxysilane, 1,3-bis[3-(trimethoxysilyl)propyl]urea, 1,3-bis[3-(trimethoxysilyl)propyl]urea, Silane coupling agents containing urea groups, such as 1-[3-(triethoxysilyl)propyl]-3-phenylurea, 1,3-bis[3-(triethoxysilyl)propyl]urea, 1-[3-(triethoxysilyl)propyl]-3,3-dioctylurea, 1-[3-(triethoxysilyl)propyl]-3,3-hexadecylurea, 1-[3-(triethoxysilyl)propyl]-3-propylurea, 3-(3-dodecylurea)propyltriethoxysilane, and N-(1-phenylethyl)-N'-[3-(triethoxysilyl)propyl]urea; etc.

[0075] As having -N(R) 2 )3 +Specific examples of the silane coupling agent of the group shown can be mentioned: N-trimethoxysilylpropyl-N,N,N-trimethylammonium chloride (N-TRIMETHOXYSILYLPROPYL-N,N,N-TRI-METHYLAMMONIUM CHLORIDE), N-trimethoxysilylpropyl-N,N,N-tri-n-butylammonium chloride (N-TRIMETHOXYSILYLPROPYL-N,N,N-TRI-n-BUTYLAMMONIUM CHLORIDE), octadecyldimethyl(3-trimethoxysilyl-propyl)ammonium chloride (OCTADECYLDIMETHYL(3-TRIMETHOXYSILYL-PROPYL)AMMONIUM CHLORIDE), N-(trimethoxysilyl ethyl) benzyl-N,N,N-trimethylammonium chloride (N-(TRIMETHOXYSILYLETHYL)BENZYL-N,N,N-TRIMETHYLAMMONIUM CHLORIDE), N,N-didecyl-N-methyl-N-(3-trimethoxysilyl-propyl)ammonium chloride (N,N-DIDECYL-N-METHYL-N-(3-TRIMETHOXYSILYL-PROPYL)AMMONIUM CHLORIDE), bis(methoxyethyl)-3-trimethoxysilylpropylammonium chloride (BIS(METHOXYETHYL)-3-TRIMETHOXYSILYLPROPYLAMMONIUM CHLORIDE), trimethoxysilylpropyl modified (polyethyleneimine) (TRIMETHOXYSILYLPROPYL MODIFIED (POLYETHYLENIMINE)), dimethoxymethylsilylpropyl modified (polyethyleneimine) (DIMETHOXYMETHYLSILYLPROPYL MODIFIED (POLYETHYLENIMINE)), and the like.

[0076] Among these, 3-aminopropyltriethoxysilane, bis(methoxyethyl)-3-trimethoxysilylpropylammonium chloride (BIS(METHOXYETHYL)-3-TRIMETHOXYSILYLPROPYLAMMONIUM CHLORIDE), trimethoxysilylpropyl modified (polyethyleneimine) (TRIMETHOXYSILYLPROPYL MODIFIED (POLYETHYLENIMINE)), N-trimethoxysilylpropyl-N,N,N-trimethylammonium chloride (N-TRIMETHOXYSILYLPROPYL-N,N,N-TRI-METHYLAMMONIUM CHLORIDE) are preferred. These have the -N(R1 )2 and / or -N(R 2 )3 + One type of silane coupling agent with the indicated group may be used, or two or more may be used in combination.

[0077] Treatment as a silane coupling agent (by having -N(R) 1 )2 and / or -N(R 2 )3 + The method for treating the substrate surface with a silane coupling agent containing the indicated group is not particularly limited, and any method capable of coating (treating) the substrate surface with a silane coupling agent can be used. Specifically, a substrate with a silane coupling agent coating (treating) its surface can be manufactured by applying (coating), spraying (spraying), or impregnating a silane coupling agent solution / dispersion obtained by dissolving / dispersing the silane coupling agent in various solvents onto the substrate surface using known methods. The coating method, spraying method, and impregnation method can be any known method.

[0078] As a solvent, any substance that the silane coupling agent can dissolve / disperse is acceptable; there are no particular limitations, as long as the appropriate silane coupling agent is selected. Examples include: water, organic solvents, and mixtures thereof. Examples of organic solvents include: alcohols such as methanol, ethanol, n-propanol, isopropanol, and methoxypropanol; ketones such as acetone and methyl ethyl ketone; tetrahydrofuran, acetonitrile, ethyl acetate, and toluene.

[0079] The concentration of the silane coupling agent solution / dispersion can be appropriately set according to the type of silane coupling agent, solvent, and treatment method (coating method, spraying method, impregnation method).

[0080] The processing conditions, such as processing temperature and processing time, for silane coupling agent treatment can be appropriately set according to the type of silane coupling agent, solvent, and processing method. However, it is preferable to perform the treatment (maintaining) under conditions of humidity of 50% or higher during the treatment of the substrate surface with the silane coupling agent solution / dispersion (coating, spraying, impregnation, etc.). This tends to strengthen the chemical bond between the substrate surface and the silane coupling agent. More preferably, the humidity is 60% or higher, and even more preferably 80% or higher. There is no particular upper limit, but for example, 100% or lower is preferred.

[0081] When processing (holding) under conditions of humidity above 50%, the processing time and temperature can be appropriately set from the viewpoints of strengthening the chemical bond between the substrate surface and the silane coupling agent, as well as from the viewpoints of economy. For example, the processing time (holding time) is preferably 1 to 60 hours, more preferably 1 to 20 hours. The processing temperature (holding temperature) is preferably 20 to 65°C, more preferably 25 to 50°C.

[0082] As the substrate whose surface is coated (treated) with a silane coupling agent, in addition to the substrate produced by the above-described method, commercially available products can also be used. As the commercially available product treated with a silane coupling agent, for example, MAS coated slide glass, MAS-GP type coated slide glass, Frontier coated slide glass, APS coated slide glass (manufactured by Matsunami Glass Industrial Co., Ltd.), and the like can be used.

[0083] From the viewpoint of the adhesion of the hydrophilic polymer layer to the surface of the substrate, at least a part (part or all) of the surface of the substrate having the group represented by -N(R 1 )2and / or -N(R 2 )3 + of the above-described method or the like preferably has a contact angle of 50 to 120 degrees with respect to water, and more preferably 70 to 100 degrees.

[0084] The above-described hydrophilic substrate is a substrate whose surface having the group represented by -N(R 1 )2and / or -N(R 2 )3 + of the above-described method or the like is formed with a hydrophilic polymer layer, and the hydrophilic polymer constituting the hydrophilic polymer layer can be appropriately selected from a substance having hydrophilicity.

[0085] As the hydrophilic polymer, for example, one or two or more kinds of homopolymers and copolymers of a hydrophilic monomer; copolymers of one or two or more kinds of hydrophilic monomers and one or two or more kinds of other monomers; and the like can be exemplified.

[0086] As the hydrophilic monomer constituting the above-described homopolymers and copolymers, various monomers having a hydrophilic group can be used. The hydrophilic group can be exemplified by, for example, amide group, sulfate group, sulfonic acid group, carboxylic acid group, hydroxyl group, amino group, amide group, ethylene oxide group, and the like.

[0087] As specific examples of the hydrophilic monomer, (meth)acrylic acid, (meth)acrylate ((meth)acrylate methoxyethyl ester and the like (meth)acrylate alkoxyalkyl ester; (meth)acrylate hydroxyalkyl ester and the like), (meth)acrylamide, (meth)acrylamide derivative having a cyclic group ((meth)acryl morpholine and the like), and the like can be exemplified. Among these, (meth)acrylic acid, (meth)acrylate, (meth)acrylate alkoxyalkyl ester, and more preferably (meth)acrylate alkoxyalkyl ester, and particularly preferably 2-methoxyethyl acrylate are preferred.

[0088] The other monomer in the above copolymer can be appropriately selected within a range not to hinder the effect of the hydrophilic polymer. For example, aromatic monomers such as styrene, vinyl acetate, N-isopropyl acrylamide, which can impart temperature responsiveness, and the like can be exemplified.

[0089] As the above homopolymer and copolymer, specifically, a homopolymer composed of one kind of hydrophilic monomer such as polyacrylic acid, polyacrylate, polymethacrylic acid, polymethacrylate, polyacrylamide, polymethacrylamide, and the like; a copolymer composed of two or more kinds of the above exemplified hydrophilic monomers; and a copolymer composed of one or more kinds of the above exemplified hydrophilic monomers and one or more kinds of the above exemplified other monomers can be exemplified. One kind of hydrophilic polymer can be used, or two or more kinds thereof can be used in combination.

[0090] Among them, as the hydrophilic polymer, at least one kind of polymer represented by the following formula (I) is preferable.

[0091] [Chemical Formula 8]

[0092]

[0093] (In the formula, R 51 represents a hydrogen atom or a methyl group, R 52 represents an alkyl group; p is 1 to 8, m is 1 to 5, and n represents the number of repetitions).

[0094] As the above polymer represented by formula (I), for example, a polymer represented by the following formula (I-1) can be appropriately used.

[0095] [Chemical Formula 9]

[0096]

[0097] (In the formula, R 51 represents a hydrogen atom or a methyl group, R 52 represents an alkyl group. m represents 1 to 5, and n represents the number of repetitions).

[0098] R 52 The number of carbon atoms of the alkyl group of R 52 is preferably 1 to 10, and more preferably 1 to 5. Among them, R 52 is particularly preferably a methyl group or an ethyl group. p is preferably 1 to 5, and more preferably 1 to 3. m is preferably 1 to 3. n (the number of repeating units) is preferably 15 to 1500, and more preferably 40 to 1200.

[0099] In addition, as the hydrophilic polymer, a copolymer of at least one kind of hydrophilic monomer selected from the compounds represented by the following formula (II) and other monomers can be appropriately used.

[0100] [Chemical Formula 10]

[0101]

[0102] (In the formula, R 51 , R 52 , p, and m are the same as described above).

[0103] As the compound represented by the above formula (II), for example, a compound represented by the following formula (II-1) can be suitably used.

[0104] [Chemical Formula 11]

[0105]

[0106] (In the formula, R 51 , R 52 , and m are the same as described above).

[0107] Among the aforementioned hydrophilic polymers, from the viewpoint of uniformity, unevenness, and the like of the hydrophilic polymer layer, the hydrophilic polymer represented by the above formula (I) is particularly suitable.

[0108] From the viewpoint of selective adsorptivity / adhesivity to cancer cells, the number average molecular weight (Mn) of the hydrophilic polymer is preferably from 8,000 to 150,000, more preferably from 10,000 to 60,000, and further preferably from 12,000 to 50,000. Note that, in the present specification, Mn can be obtained from a measured value based on gel permeation chromatography (GPC) (GPC-8000 series manufactured by Tosoh Corporation, detector: differential refractometer, column: TSKGEL SUPERMULTIPORE HZ-M manufactured by Tosoh Corporation) and conversion to a standard polystyrene.

[0109] The film thickness of the hydrophilic polymer layer (layer formed of a hydrophilic polymer) is preferably from 10 to 1,000 nm, more preferably from 30 to 700 nm, and further preferably from 50 to 400 nm. By adjusting to the above range, good low adsorptivity to proteins or cells and selective capturing of cancer cells can be obtained.

[0110] At least a part (part or all) of the surface of the hydrophilic polymer layer (surface of the hydrophilic polymer layer of the aforementioned hydrophilic substrate) preferably has a contact angle with water of from 20 degrees to 70 degrees or less, and more preferably from 30 degrees to 60 degrees or less.

[0111] The surface of the hydrophilic polymer layer (surface of the hydrophilic polymer layer of the aforementioned hydrophilic substrate) preferably has a surface roughness Ra of 0.30 μm or less, more preferably 0.20 μm or less, and further preferably 0.18 μm or less. The lower limit is not particularly limited, and the smaller the better.

[0112] Note that in the present specification, the surface roughness Ra is the center line average surface roughness Ra defined in JIS B 0601-2001.

[0113] The hydrophilic polymer solution / dispersion liquid obtained by (1) dissolving / dispersing the hydrophilic polymer in various solvents is injected to the surface of the substrate (substrate recess, etc.) having a group represented by -N(R 1 )2 and / or -N(R 2 )3 + The hydrophilic polymer solution / dispersion liquid is applied (sprayed) to the surface of the substrate, and the substrate is left for a predetermined time and dried, whereby a substrate having a hydrophilic polymer layer formed of the hydrophilic polymer is produced. Further, by adding other components to the substrate having the hydrophilic polymer layer as necessary, a device capable of capturing, culturing, and inspecting specific cells, etc. can be produced.

[0114] The solvent injection method, the application (spraying) method, etc. can use conventionally known materials and methods. The leaving and drying times of (1) and (2) can be appropriately set according to the size of the substrate, the type of the liquid to be introduced, etc. The leaving time is preferably 5 minutes to 10 hours, more preferably 10 minutes to 5 hours, and further preferably 15 minutes to 2 hours. The drying is preferably performed at room temperature (about 23°C) to 80°C, and more preferably at room temperature to 50°C. In addition, reduced pressure drying can be performed. Further, the excess hydrophilic polymer solution / dispersion liquid can be appropriately drained after leaving for a certain time, and dried.

[0115] As the solvent, any solvent that can dissolve the hydrophilic polymer can be used without particular limitation, and can be appropriately selected according to the hydrophilic polymer to be used. For example, the solvent that can be used in the dissolution / dispersion of the silane coupling agent described above can be used.

[0116] The hydrophilic polymer layer can also be a substance having fibronectin adsorbed to the surface of the hydrophilic polymer layer. By further adsorbing fibronectin to the surface of the hydrophilic polymer layer, the capturing property of specific cells such as cancer cells can be improved.

[0117] The method of adsorbing fibronectin to the hydrophilic polymer layer is not particularly limited, and a publicly known method can be used. For example, by contacting a buffered solution (phosphate buffered saline (PBS), etc.) of fibronectin with the hydrophilic polymer layer using a publicly known method, leaving it at a prescribed temperature for a prescribed period of time, and performing washing as necessary, etc., fibronectin can be adsorbed. The temperature and time can be suitably set, and for example, it can be performed at 10 to 60°C for 0.1 to 24 hours.

[0118] From the viewpoint of adsorbing fibronectin to the hydrophilic polymer layer, it is preferable to use a solution, dispersion, etc. in which the concentration of fibronectin is preferably adjusted to 0.5 to 500 μg / ml, and more preferably adjusted to 1 to 250 μg / ml. By adjusting the concentration to the above range, superior capturing properties for specific cells such as cancer cells can be obtained.

[0119] The hydrophilic substrate production method of the present application is a method of forming a hydrophilic polymer layer on a substrate surface having a group represented by -N(R 1 )2and / or -N(R 2 )3 + (R 1 and R 2 are each the same or different, and are a hydrogen atom or a substituted or unsubstituted hydrocarbon group which can contain a hetero atom). As described above, first, by performing an introduction treatment of a group represented by -N(R 1 )2and / or -N(R 2 )3 + on the substrate surface, and then forming a hydrophilic polymer layer on the surface to which the group has been introduced, a hydrophilic polymer layer having controlled uniformity of the surface, controlled unevenness of the surface, small surface roughness, and high smoothness can be formed.

[0120] When the above hydrophilic substrate production method is used, for example, by contacting a sample (blood, body fluid, etc.) with a hydrophilic substrate on which a hydrophilic polymer layer has been formed on the substrate surface after the introduction of a group represented by -N(R 1 )2and / or -N(R 2 )3 + , it is expected that the capturing properties of specific cells will be improved, and the unevenness of the capturing properties will be suppressed. Furthermore, by measuring the number of captured specific cells, the number of specific cells in the collected blood or body fluid can be known, and it is expected that the confirmation of the effects of cancer treatment, etc. will be possible.

[0121] [Examples]

[0122] The present application will be specifically described below based on examples, but the present application is not limited only to this.

[0123] (Production of hydrophilic polymer)

[0124] Poly(2-methoxyethyl acrylate) (PMEA) was produced by thermal polymerization of 2-methoxyethyl acrylate (25 wt% in toluene) at 60°C for 7 hours using AIBN (azobisisobutyronitrile) 12.5 mg / ml in toluene.

[0125] (Example 1)

[0126] A 1N aqueous NaOH solution was added to the chambered slide (bottom surface: soda lime glass) and left for 1 hour. After that, the chamber surface was washed with water and naturally dried. Next, a 0.5% concentration of 3-aminopropyltriethoxysilane (Tokyo Chemical Industry Co., Ltd. A0439) solution (mixed solution of methanol / water = 50 / 50) was added to the chambered slide, left for 5 hours at 80% humidity and 40°C, and dried under reduced pressure at 60°C. Next, water washing and drying were performed, and a 0.25% methanol solution of the produced PMEA was injected into the chambered slide. After that, vacuum drying was immediately performed in an oven at 40°C for 5 minutes to produce a hydrophilic substrate.

[0127] (Example 2)

[0128] The silane coupling agent was changed from 3-aminopropyltriethoxysilane to bis(methoxyethyl)-3-trimethoxysilylpropylammonium chloride (SIB1500 manufactured by Gelest, the following formula), and otherwise the same operation as Example 1 was performed to produce a hydrophilic substrate.

[0129] [Chemical Formula 12]

[0130]

[0131] (Example 3)

[0132] The silane coupling agent was changed from 3-aminopropyltriethoxysilane to trimethoxysilylpropyl modified (polyethylenimine) (SSP-060 manufactured by Gelest, the following formula, Mn 1500-1800), and otherwise the same operation as Example 1 was performed to produce a hydrophilic substrate.

[0133] [Chemical Formula 13]

[0134]

[0135] (Example 4)

[0136] A 0.5% concentration of 3-aminopropyltriethoxysilane (Tokyo Chemical Industry Co., Ltd. A0439) solution (mixed solution of methanol / water = 50 / 50) was added to the chambered slide, left for 5 hours at 80% humidity and 40°C, and dried under reduced pressure at 60°C. Next, water washing and drying were performed, and a 0.25% methanol solution of the produced PMEA was injected into the chambered slide. After that, vacuum drying was immediately performed in an oven at 40°C for 5 minutes to produce a hydrophilic substrate.+ A commercially available chambered glass slide (manufactured by Matsunami Glass Industry Co., Ltd., MAScoated type) was used instead of a standard chambered glass slide, into which a 0.25% methanol solution of the prepared PMEA was injected. Subsequently, the slide was immediately vacuum-dried in an oven at 40°C for 5 minutes to produce a hydrophilic substrate.

[0137] (Example 5)

[0138] In Example 4, after vacuum drying in an oven at 40°C for 5 minutes, fibronectin was further adsorbed. Specifically, a 200 μg / ml PBS solution (phosphate-buffered saline) containing fibronectin was prepared and injected, and after being placed at 40°C for 1 hour, it was washed with PBS solution to prepare a hydrophilic substrate.

[0139] (Comparative Example 1)

[0140] A 0.25% methanol solution of the prepared PMEA was injected into the untreated slide chamber. Immediately afterwards, it was vacuum dried in an oven at 40°C for 5 minutes to prepare a hydrophilic substrate.

[0141] [Water contact angle (with -N(R)] 1 )2-base or -N(R 2 )3 + [Substrate surface of base]

[0142] Importing -N(R) 1 )2 or -N(R 2 )3 + Two μl of distilled water was dropped onto the surface of the substrate with the indicated group, and the contact angle was measured after 30 seconds using the θ / 2 method (room temperature).

[0143] In addition, the contact angle of the untreated slide was measured in Comparative Example 1.

[0144] [Film thickness of the hydrophilic polymer layer (coating layer)]

[0145] Regarding the film thickness of the hydrophilic polymer layer on the hydrophilic substrate, TEM was used to measure (photograph) the cross-section of the hydrophilic polymer layer by accelerating the voltage to 15kV, 1000 times.

[0146] [Surface roughness Ra (hydrophilic polymer layer)]

[0147] Using a laser microscope, the surface roughness at four points (the first peak) on a hydrophilic substrate (hydrophilic polymer layer) was measured non-contactly, and the average value of the four points Ra was taken as the surface roughness Ra (the average value of the centerline surface roughness Ra specified in JIS B0601-2001).

[0148] [Table 1]

[0149]

[0150] In Figure 1 Fig. 1 shows a photograph of the coated substrate prepared in Example 1.

[0151] In Comparative Example 1, while the entire surface was observed to be slightly hazy, portions where white spots were observed. When poly(2-methoxyethyl acrylate) was prepared several times in the same manner as in Comparative Example 1, the haze was sometimes observed in a ring shape, or in a spot shape, but portions where haze was observed were often observed.

[0152] On the other hand, in Examples 4 and 5, in which the substrate having a surface with a -N(R 1 )2group or a -N(R 2 )3 + group was used, a coated film was formed that was not hazy (Example 4) or was very slightly hazy (Example 5), and the uniformity and unevenness of the surface were controlled. The portions where haze was observed reflected the aggregation state of the poly(2-methoxyethyl acrylate) and the surface unevenness, and it was easy to imagine that the capture performance of specific cells that were different from the other smooth portions was exhibited in these portions, and that this was the cause of the uneven capture performance. Therefore, it was expected that a surface that was not hazy, in which the uniformity and unevenness were controlled (Ra 0.30 or less), would exhibit a capture performance with less unevenness.

Claims

1. A hydrophilic substrate, which has a hydrophilic polymer layer formed on the substrate surface with a group represented by the following formula (1-2) introduced to the substrate surface by treating the substrate surface with a silane coupling agent, -N(R1)2 (1-2) 2 )3 + wherein, R 2 respectively identical or different, are a hydrogen atom, or a substituted or non- substituted hydrocarbon group, with or without heteroatoms, wherein R 11 ~R 13 are the same or different, a hydrogen atom, or a substituted or unsubstituted monovalent hydrocarbon group with or without heteroatoms; R 16 ~R 18 are the same or different, a substituted or unsubstituted monovalent hydrocarbon group with or without heteroatoms; R 19 is a substituted or unsubstituted divalent hydrocarbon group with or without heteroatoms; X is halogen.

2. The hydrophilic substrate according to claim 1, wherein, The hydrophilic polymer layer is a layer formed using a hydrophilic polymer solution and / or a hydrophilic polymer dispersion.

3. The hydrophilic substrate according to claim 1, wherein, The hydrophilic polymer layer is formed of at least one hydrophilic polymer selected from a polymer represented by the following formula (I), wherein R 51 represents a hydrogen atom or a methyl group, R 52 represents an alkyl group; p represents 1 to 8, m represents 1 to 5, and n represents the number of repetitions.

4. The hydrophilic substrate according to claim 1, wherein, The hydrophilic polymer layer is formed of a copolymer of at least one hydrophilic monomer selected from a compound represented by the following formula (II) and other monomers, wherein R 51 represents a hydrogen atom or a methyl group, R 52 represents an alkyl group, p represents 1 to 8, and m represents 1 to 5.

5. The hydrophilic substrate according to claim 1, wherein, The thickness of the hydrophilic polymer layer is 10 to 1000 nm.

6. The hydrophilic substrate according to claim 1, wherein, Fibronectin is adsorbed on the surface of the hydrophilic polymer layer.

7. The hydrophilic substrate according to claim 1, wherein, The silane coupling agent is bis(methoxyethyl)-3-trimethoxysilylpropyl ammonium chloride, trimethoxysilylpropyl modified (polyethyleneimine), or N-trimethoxysilylpropyl-N,N,N-trimethylammonium chloride.

8. A method for producing a hydrophilic substrate, in which a hydrophilic polymer layer is formed on a substrate surface having a group represented by -N(R1)2 introduced by treating the substrate surface with a silane coupling agent represented by the following formula (1-2) ###0002### (1-2) 2 )3 + represented by -N(R1)2 introduced by treating the substrate surface with a silane coupling agent represented by the following formula (1-2) ###0003### (1-2) R 2 each independently of one another or of one another, a hydrogen atom, or a substituted or unsubstituted hydrocarbon group, with or without heteroatoms, wherein R 11 ~R 13 are the same or different, a hydrogen atom, or a substituted or unsubstituted monovalent hydrocarbon group with or without heteroatoms; R 16 ~R 18 are the same or different, a substituted or unsubstituted monovalent hydrocarbon group with or without heteroatoms; R 19 is a substituted or unsubstituted divalent hydrocarbon group with or without heteroatoms; X is halogen.

9. The method of making a hydrophilic substrate according to claim 8, wherein, The hydrophilic polymer layer is a layer formed using a hydrophilic polymer solution and / or a hydrophilic polymer dispersion.

10. The method of making a hydrophilic substrate according to claim 8, wherein, The hydrophilic polymer layer is formed of at least one hydrophilic polymer selected from a polymer represented by the following formula (I), wherein R 51 represents a hydrogen atom or a methyl group, R 52 represents an alkyl group, p is 1 to 8, m is 1 to 5, and n represents the number of repetitions.

11. The method for producing a hydrophilic substrate according to claim 8, wherein the hydrophilic polymer layer is formed of a copolymer of at least one hydrophilic monomer selected from a compound represented by the following formula (II) and other monomers, wherein R 51 represents a hydrogen atom or a methyl group, R 52 represents an alkyl group, p represents 1 to 8, and m represents 1 to 5.

12. The method of making a hydrophilic substrate according to claim 8, wherein, The thickness of the hydrophilic polymer layer is 10 to 1000 nm.

13. The method of making a hydrophilic substrate according to claim 8, wherein, Fibronectin is adsorbed on the surface of the hydrophilic polymer layer.

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