JP2016112397A - Member for cell culture - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a member for cell culture that has a low adsorptivity of plasma protein, hemolyticity and cytotoxicity.SOLUTION: Provided is a member for cell culture (4) including a member for cell culture (3), and a coating for cell culture placed on the surface of the member for cell culture (3), and in which said coating for cell culture comprises a block polymer (A) obtained by bonding with a particular bonding chain between an acrylic polymer moiety (A1) formed from an acrylic monomer having an average value of water/1-octanol distribution coefficient (LogP) of 0 or more and 2 or less, and a polyethylene oxide moiety (A2).SELECTED DRAWING: None

Description

  The present invention relates to a cell culturing member using a novel biocompatible resin.

  Conventionally, polymer materials used in many medical devices such as blood bags and extracorporeal circulation circuits for cardiopulmonary bypass have no blood compatibility, so the use of anticoagulants is indispensable when using medical devices. there were. In addition, when biological components such as blood cells and proteins are adsorbed and denatured on the surface of the polymer material, blood coagulation is caused, causing not only adverse effects such as thrombus formation and inflammatory reaction to the living body, but also deterioration of the material. This is one of the important issues that must be solved in medical materials. In this way, with the advancement of medical technology, the opportunity for polymer materials to come into contact with living tissue or blood is increasing, and the blood compatibility of polymer materials themselves has become a major issue. .

  In the development of cell culture materials, which are one of the tools for drug discovery research and regenerative medicine research, it is generally known that collagen, polystyrene and polymethyl methacrylate are good. It is superior in low cytotoxicity, economical efficiency, and processability, and in the current tissue culture, polystyrene hydrophilized is used. However, in such a culture member, although cell growth and proliferation are observed, it is impossible to form a three-dimensional cell structure that is advantageous for maintaining the function of the cell. There is a need for polymeric materials.

  Against this background, many surface design methods have been studied with the aim of realizing excellent biocompatible polymer materials.

  As the surface of the biocompatible polymer, effectiveness such as a microphase separation surface, a hydrophilic surface, and a surface structure similar to a cell membrane containing a zwitterionic molecule has been reported. For example, a polymer having 2-methacryloyloxyethyl phosphorylcholine (MPC) having a phosphobetaine-type phospholipid polar group as a component can obtain a superhydrophilic surface when treated on the surface of the substrate. Since cells such as spheres do not adhere at all, they are used as surface treatment agents for various medical products such as heart-lung machines, stents, contact lenses, and cell culture substrates. Although 2-methoxyethyl acrylate has a simple chemical structure, it is a polymer material that exhibits biocompatibility in the same way as MPC polymer, so it can be used as a surface treatment agent for many medical devices such as heart-lung machines. It has become.

  As other biocompatible materials, many resins incorporating a polyethylene oxide moiety (hereinafter sometimes abbreviated as PEG) have been studied. PEG has high hydrophilicity and low antigenicity, and has been conventionally used as a nonionic surfactant, a plasticizer, a pharmaceutical base material, and the like. However, while PEG has excellent blood compatibility, it is water-soluble, so when used as a medical material, it needs to be used as a block copolymer or graft copolymer with other polymers. is there.

  In order to meet such a demand, use of a polymer in which a PEG chain is incorporated into a main chain or a side chain has been proposed. For example, as a polymer in which a PEG chain is incorporated into a main chain, Patent Document 1 discloses the use of a PEG-containing polyester, and Patent Document 2 discloses the use of a PEG-containing liquid crystalline polyester. Patent Document 3 discloses the use of PEG-containing polyamide, Patent Document 4 discloses the use of PEG-containing polyimide, and Patent Document 5 discloses a PEG copolymer having an alkyl chain as a side chain. As a polymer in which a PEG chain is incorporated in a side chain, an acrylic resin as disclosed in Patent Document 6 has been proposed. Furthermore, Patent Document 7 discloses an acrylic resin in which a PEG chain is incorporated in the main chain as in the present invention, but there is no description regarding biocompatibility taken up in the present invention.

JP-A-9-290019 JP-A-11-111050 JP-A-9-302223 JP-A-6-14989 JP 2005-281665 A WO2004 / 87228 JP 2007-77292 A

  An object of the present invention is to provide a cell culture member having low plasma protein adsorptivity, hemolysis, and low cytotoxicity.

  In general, PEG is considered to cause thrombus formation by promoting fibrin formation by inducing complement activation at the time of blood contact due to high mobility and free hydroxyl group that interacts strongly with polar groups in the living body. It is not appropriate to use it for important medical materials. Therefore, the present inventors reduced the number of hydroxyl groups present at the end of PEG and excessive mobility by incorporating PEG into the main chain of the block copolymer, and maintained the hydrophilicity and flexibility of PEG. However, a polymer with excellent biocompatibility, in which the problem of thrombus formation was improved, was investigated. As a result, a block copolymer having an acrylic polymer part (A1) and a polyethylene oxide part (A2) using an acrylic monomer having an average value of water / 1-octanol partition coefficient (LogP) of 0 or more and 2 or less. When forming a coalescence, the block copolymer has been found to exhibit good blood compatibility and biocompatibility while maintaining the properties of PEG, and can further improve the workability during coating. Arrived.

That is, the present invention is a cell culture member (4) comprising a cell culture member (3) and a cell culture coating film positioned on the surface of the cell culture member (3),
The cell culture coating film,
An acrylic polymer portion (A1) formed from an acrylic monomer having an average value of water / 1-octanol partition coefficient (LogP) of 0 or more and 2 or less, and a polyethylene oxide portion (A2) are represented by the following formula (I And a block polymer (A) bonded with
The present invention relates to a cell culture member (4).

  In the said invention, the mass of a polyethylene oxide part (A2) contained in a block polymer (A) / [total mass of an acrylic polymer part (A1) and a polyethylene oxide part (A2)] = 0.01-0 .5 is preferable.

  In the invention, the block polymer (A) is a polyethylene oxide moiety (A2) represented by the following formula (II), wherein a monomer having an average water / 1-octanol partition coefficient (LogP) of 0 or more and 2 or less. And a block copolymer obtained by polymerization using a polymer azo polymerization initiator having a mass average molecular weight of 5,000 to 100,000.

In the formula, m and n represent an integer of 1 or more.

  Furthermore, in the said invention, an acrylic polymer part (A1) is a homopolymer part or a copolymer part formed only from the acrylic monomer whose water / 1-octanol partition coefficient (LogP) is 0 or more and 2 or less. Is preferred.

In addition, the present invention provides a cell culture member (3) having a surface,
An acrylic polymer portion (A1) formed from an acrylic monomer having an average value of water / 1-octanol partition coefficient (LogP) of 0 or more and 2 or less, and a polyethylene oxide portion (A2) are represented by the following formula (I A coating agent containing the block polymer (A) formed by bonding with a), and then dried or cured to provide a cell culture coating (4). Regarding the method.

The block polymer (A) mainly composed of PEG and a specific acrylic polymer part has low plasma protein adsorptivity, hemolysis and cytotoxicity, and has good processability when applied. It is an excellent biocompatible resin.
The biocompatible resin can be applied to a cell culture member, which is one of the tools for drug discovery research and regenerative medicine research. The biocompatible resin may be any disposable medical device used in combination with an anticoagulant, such as a vacuum blood collection tube, blood bag, extracorporeal circuit, medical device such as an oxygenator, a stent, and the like. Can be suitably used for medical devices that come into contact with blood, such as artificial organs.

<Block polymer (A)>
The block polymer (A) is an acrylic polymer part (A1) formed from a monomer having an average value of water / 1-octanol partition coefficient (cLogPow: hereinafter referred to as LogP) of 0 or more and 2 or less (hereinafter referred to as 0 to 2). ) And a polyethylene oxide part (A2).

  LogP is one of the numerical values representing the properties of the chemical substance, and is a constant value that does not depend on the addition amount. The target substance is a mixture of water and 1-octanol, and the concentration of each phase is shown in the common logarithm for the case where the aqueous phase and the octanol phase are in an equilibrium state in contact with each other. . When LogP increases, the hydrophobicity tends to increase relatively, and when LogP decreases, the hydrophilicity tends to increase relatively.

  The measurement of LogP can be generally performed by a flask immersion method described in JIS Japanese Industrial Standard Z7260-107 (2000). In addition, LogP can be estimated by a computational chemical method or an empirical method instead of actual measurement. As a method and software used for calculating LogP, publicly known methods can be used, but in the present invention, LogP is obtained by using a program incorporated in the system: ChemdrawPro 11.0 of CambridgeSoft.

  The block polymer (A) of the present invention is an acrylic polymer portion formed from a monomer having an average value of LogP of 0 or more and 2 or less (hereinafter, 0 to 2) as compared with a conventional PEG polymer. Therefore, solubility in water is controlled and fat solubility is improved. As a result, when applied to a medical device that comes into contact with blood or living tissue, coating on the medical device proceeds rapidly. Furthermore, a crosslinked coating film can also be formed by copolymerizing a monomer having a functional group capable of reacting with a crosslinking agent described later on the acrylic polymer portion (A1) in the block polymer (A). By forming the coating film, peeling and elution of the block polymer (A) can be suppressed. Examples of the functional group that the acrylic polymer portion (A1) may have include a carboxyl group, a hydroxyl group, an epoxy group, an amino group, and an isocyanate group.

Moreover, since the block polymer (A) of the present invention incorporates a polyethylene oxide moiety in the main chain with respect to a conventional acrylic polymer, microphase separation is induced, resulting in a low antigen due to a change in the surface aggregate structure. Sex can be imparted.
As described above, the block polymer (A) of the present invention has a molecular weight, a side chain functional group, and a plasma protein adsorption property, hemolysis property, cytotoxicity control and biological safety. It is possible to optimize the group type and side chain functional group introduction amount according to the purpose of use.

Due to the balance between the highly hydrophilic polyethylene oxide part (A2) and the hydrophobic acrylic polymer part (A1) constituting the block polymer (A), it is possible to suppress plasma protein adsorption, hemolysis, cytotoxicity, etc. It can be controlled effectively.
When the block polymer (A) of the present invention is used as a coating material, the amount and physical properties of the acrylic polymer portion (A1) are controlled in order to improve the wettability to the base material, that is, the medical device (1) described later. It is possible. By introducing the acrylic polymer portion (A1) suitable for the properties of the base material, it is possible to sufficiently exhibit the target plasma protein adsorptive properties, hemolytic properties, and cytotoxicity suppression effects. The block polymer (A) has a mass of polyethylene oxide part (A2) / [total mass of acrylic polymer part (A1) and polyethylene oxide part (A2)] = 0.01 to 0.5. Preferably, (A2) / [(A1) + (A2)] = 0.05 to 0.3 is more preferable.

  The block polymer (A) is polymerized using a polymer azo polymerization initiator having a polyethylene oxide part (A2) represented by the following formula (II) and having a mass average molecular weight of 5,000 to 100,000. It is preferable to obtain it.

(In the formula, m and n represent an integer of 1 or more.)

The mass average molecular weight of the polymer azo polymerization initiator may be about 5000 to 100,000, preferably 10,000 to 50,000. The molecular weight of the PEG moiety of the initiator may be about 800 to 10,000, preferably about 1000 to 8000.
The polymer azo polymerization initiator includes a polyethylene oxide part (A2) and a repeating part containing an azo group (—N═N—).
Since the polymer azo polymerization initiator has a polyethylene oxide part (A2), it is soluble in water, alcohol, and organic solvent, and the block polymer (A) is formed by solution polymerization, emulsion polymerization, or dispersion polymerization. Synthesis is possible. In addition, since it has a polymerization initiation part (radical generation part: -N = N-) in the molecular chain skeleton, it is not necessary to use a separate polymerization initiator, and more radically compared to the terminal reactive macromonomer. It is characterized by high reactivity and stability.
The polymer azo polymerization initiator is: C (CH ₃ ) CN— (CH ₂ ) ₂ —COO— (CH ₂ CH ₂ O) _m —CO— (CH ₂ ) ₂ —C (CH ₃ ) CN A radical as shown below is generated, and an acrylic monomer described later is polymerized. And the main chain which the acrylic polymer part (A1) formed from an acrylic monomer and the part derived from the said radical couple | bonded is formed, and a block polymer (A) is formed. The polyethylene oxide part (A2) is derived from a part of the radical.
Specific examples of the polymer azo polymerization initiator include a polymer azo initiator VPE0201 manufactured by Wako Pure Chemical Industries, Ltd. (the molecular weight of the (CH ₂ CH ₂ O) _m portion is about 2000, and n is about 6). .

  In the present invention, 2,2′-azobisisobutyronitrile, 2,2′-azobis (2-methylbutyronitrile), 1,1′- is used in combination with the above-mentioned polymer azo polymerization initiator. Azobis (cyclohexane 1-carbonitrile), 2,2′-azobis (2,4-dimethylvaleronitrile), 2,2′-azobis (2,4-dimethyl-4-methoxyvaleronitrile), dimethyl 2,2 ′ -Azobis (2-methylpropionate), 4,4'-azobis (4-cyanovaleric acid), 2,2'-azobis (2-hydroxymethylpropionitrile), 2,2'-azobis [2 -(2-imidazolin-2-yl) propane] and the like, benzoyl peroxide, tert-butyl perbenzoate, cumene hydroperoxide and diisopropyl peroxydi -Bonate, di-n-propyl peroxydicarbonate and di (2-ethoxyethyl) peroxydicarbonate, tert-butylperoxy-2-ethylhexanoate, tert-butylperoxyneodecanoate and tert-butyl Organic peroxide initiators such as peroxybivalate, (3,5,5-trimethylhexanoyl) peroxide, dipropionyl peroxide and diacetyl peroxide can be used. By using these initiators together, the starting efficiency can be increased and the acrylic polymer portion (A1) can be efficiently incorporated into PEG, and the residual monomer can be reduced.

  Moreover, at the time of synthesis | combination, you may control molecular weight and terminal structure using chain transfer agents, such as mercaptans, such as lauryl mercaptan and n-dodecyl mercaptan, (alpha) -methylstyrene dimer, limonene according to a use.

<Monomer>
Next, an acrylic monomer that is a raw material for the acrylic polymer portion (A1) of the block polymer (A) will be described. In the present invention, the acrylic monomer means both an acrylic monomer and a methacrylic monomer.
From the viewpoint of biocompatibility of the formed block polymer (A), the average value of the water / 1-octanol partition coefficient (LogP) of the acrylic monomer is 0 or more and 2 or less. When synthesizing using an acrylic monomer having an average value of LogP lower than 0, the hydrophilicity of the finally obtained block copolymer is increased and the solubility in water is improved, so that a stable coating film is formed. I can't.
On the other hand, when synthesizing using an acrylic monomer having an average value of LogP higher than 2, although the solubility of the block copolymer finally obtained in water can be suppressed, the hydrophobicity increases too much, so protein adsorption Can not be suppressed. Furthermore, LogP has a high correlation with biological activity (toxicity) and maximizes the effects of protein / cell adsorption, platelet adhesion / activation, and biological safety, which are the subject of the present invention. Can not be demonstrated.

  A method for obtaining the average value of LogP in the present invention will be described. In the present invention, the average value of LogP is a value obtained by averaging LogP of each monomer used by the weight% of each monomer. That is, when a monomer having LogP of 0 and a monomer having LogP of 2 are charged at a ratio of 50:50 wt%, the average value of LogP is 1.

  In this invention, if LogP of the acryl-type monomer to be used is the range of 0-2, it can be used as a raw material of a homopolymer part. Moreover, even if LogP of the acrylic monomer to be used is outside the range of 0 to 2, if the average value of LogP including other acrylic monomers is in the range of 0 to 2, it is used as a raw material for the copolymer portion. be able to.

Examples of the monomer having a water / 1-octanol distribution coefficient (LogP) of 0 or more and 2 or less include, for example, a monomer having no functional group serving as a crosslinking point and a monomer having a functional group serving as a crosslinking point. Can do.
As a monomer having no functional group that becomes a crosslinking point, for example,
An alkyl acrylate having 1 to 4 carbon atoms in the alkyl group;
An alkyl methacrylate having 1 to 3 carbon atoms in the alkyl group;
Methoxymethyl (meth) acrylate, 2-methoxyethyl (meth) acrylate, ethoxymethyl (meth) acrylate, ethoxyethyl (meth) acrylate, propoxymethyl (meth) acrylate, propoxyethyl (meth) acrylate, 2- (2-methoxy Ethoxy) ethyl (meth) acrylate, 2- [2- (2-methoxyethoxy) ethoxy] ethyl (meth) acrylate, 2- (2-ethoxyethoxy) ethyl (meth) acrylate, 2- [2- (2-ethoxy) Alkoxy group-containing monomers such as ethoxy) ethoxy] ethyl (meth) acrylate, tetrahydrofurfuryl (meth) acrylate;
Vinyl group-containing monomers such as (meth) acrylonitrile, vinyl acetate, butadiene, isoprene;
Amide group-containing monomers such as N-vinylpyrrolidone, N-vinyl-ε-caprolactam, 1-vinylimidazole, N-isopropylacrylamide, N, N-dimethylacrylamide;
Etc.
These may be used alone or in combination of two or more. Of these compounds, 2-methoxyethyl acrylate and tetrahydrofurfuryl acrylate are particularly preferable from the viewpoints of economy and operability.

As the monomer having a functional group that becomes a crosslinking point, a carboxyl group-containing monomer, a hydroxyl group-containing monomer, an epoxy group-containing monomer, an amino group-containing monomer, an isocyanate group-containing monomer, and the like can be used.
For example, a copolymer having a carboxyl group introduced therein can be crosslinked with an epoxy compound, an aziridine compound, a carbodiimide compound, a metal chelate compound, or an N-hydroxyethylacrylamide compound. A copolymer having a hydroxyl group introduced can be crosslinked with an isocyanate compound, a carbodiimide compound, or the like. A copolymer having an amino group introduced therein can be crosslinked with an epoxy compound. A copolymer having an isocyanate group introduced therein can be crosslinked with a hydroxyl group-containing compound. The amount of the monomer having a functional group serving as a crosslinking point is preferably 10% by mass or less in a total of 100% by mass of all monomers. By using it at 10 mass% or less, when a crosslinking agent is used together, a coating film having an appropriate crosslinking density can be obtained.

  The carboxyl group-containing monomer is not particularly limited as long as it has a carboxyl group in its structure. For example, (meth) acrylic acid, 2-carboxyethyl (meth) acrylate, or alkylene such as ethylene oxide or propylene oxide Examples include alkylene oxide addition succinic acid (meth) acrylate having a carboxyl group at the terminal where oxide is repeatedly added.

  The hydroxyl group-containing monomer is not particularly limited as long as it has a hydroxyl group in its structure. For example, 2-hydroxyethyl (meth) acrylate, 1-hydroxypropyl (meth) acrylate, (meth) acrylic acid 2-hydroxypropyl, 3-hydroxypropyl (meth) acrylate, 1-hydroxybutyl (meth) acrylate, glycerol monofunctional (meth) acrylate, polylactones having a hydroxyl group at the terminal by ring-opening addition of a lactone ring (meta ) An alkylene oxide addition system (meth) acrylate ester and glucose ring system (meth) acrylate ester having a hydroxyl group at the terminal to which an alkylene oxide such as an acrylic ester, ethylene oxide or propylene oxide is repeatedly added.

  The epoxy group-containing monomer is not particularly limited as long as it has an epoxy group in its structure, and examples thereof include glycidyl (meth) acrylate and 3,4-epoxycyclohexylmethyl (meth) acrylate. .

  The amino group-containing monomer is not particularly limited as long as it has an amino group in its structure. For example, monomethylaminoethyl (meth) acrylate, monoethylaminoethyl (meth) acrylate, (meth) acrylic Examples thereof include monomethylaminopropyl acid and monoethylaminopropyl (meth) acrylate.

  In this invention, it is preferable to copolymerize the monomer which has a functional group used as a crosslinking point from a viewpoint of suppressing peeling and elution of a block polymer (A). Thereby, the functional group which can react with the crosslinking agent mentioned later can be introduce | transduced, and a crosslinked coating film can be formed.

In the present invention, the monomer having the above-mentioned log P outside the range of 0 to 2 can be copolymerized as the monomer, but the acrylic polymer portion (A1) has a water / 1-octanol distribution coefficient (Log P) of 0. More preferably, it is a homopolymer part or a copolymer part formed only from the monomer of ~ 2.
That is, a single copolymer having a partition coefficient LogP of 0 to 2 may be used, and a block copolymer may be obtained by polymerization using the polymer azo polymerization initiator, or the partition coefficient LogP may be A block copolymer may be obtained by using a plurality of monomers of 0 to 2 and polymerizing using the above polymer azo polymerization initiator.

  Among the monomers having a LogP outside the range of 0 to 2, examples of the monomer having no functional group serving as a crosslinking point include acrylates having 5 to 20 carbon atoms in the alkyl group, and 4 to 4 carbon atoms in the alkyl group. And vinyl group-containing monomers such as 20 methacrylates and styrene.

  Among monomers having a LogP outside the range of 0 to 2, examples of the monomer having a functional group serving as a crosslinking point include carboxyl group-containing monomers such as maleic acid, 4-hydroxystyrene, and N-hydroxyethyl (meth) acrylamide. And other hydroxyl group-containing monomers.

Furthermore, in the present invention, a polyfunctional monomer may be further copolymerized in order to partially introduce a crosslinked structure into the block polymer (A).
In the case of crosslinking while copolymerizing, the crosslinking ratio increases as the amount of the polyfunctional monomer used increases, and the possibility of gelation during the reaction increases. Therefore, as a quantity of a polyfunctional monomer, the range of 0.01-10 mass% is preferable in 100 mass% of all monomers, and 0.1-5 mass% is more preferable. When the block polymer (A) is crosslinked with a polyfunctional monomer, it becomes hardly water-soluble, so that the problem of peeling off from the substrate can be reduced as in the case of using a crosslinking agent.

The polyfunctional monomer is not particularly limited as long as it has two or more ethylenically unsaturated groups in its structure. For example, ethylene glycol di (meth) acrylate, diethylene glycol di (meth) acrylate, polyethylene oxide di (Meth) acrylate, propylene glycol polyethylene oxide di (meth) acrylate, dipropylene glycol polyethylene oxide di (meth) acrylate, polypropylene glycol di (meth) acrylate, 1,6-hexanediol di (meth) acrylate, glycerin di ( (Meth) acrylate, dimethylol tricyclodecane di (meth) acrylate, tripropylene glycol di (meth) acrylate, or neopentyl glycol modified trimethylolpropane di (meth) Difunctional (meth) acrylates such as acrylate;
Trifunctional or more polyfunctional compounds such as trimethylolpropane tri (meth) acrylate, pentaerythritol tri (meth) acrylate, pentaerythritol tetra (meth) acrylate, dipentaerythritol hexa (meth) acrylate, or dipentaerythritol penta (meth) acrylate Functional (meth) acrylates; or
(Meth) acrylic acid adduct of 1,6-hexanediol diglycidyl ether, (meth) acrylic acid adduct of neopentyl glycol diglycidyl ether, (meth) acrylic acid adduct of glycerol diglycidyl ether, bisphenol A diglycidyl Examples include (meth) acrylic acid adducts of ether, (meth) acrylic acid adducts of bisphenol F type epoxy, and (meth) acrylic acid adducts of novolac type epoxy. Moreover, what modified | denatured the (meth) acrylate mentioned above further by (poly) alkylene oxide, (poly) caprolactone, etc. can also be used.

  In the present invention, the block polymer (A) can be synthesized by a known method. For example, solution polymerization, bulk polymerization, emulsion polymerization, dispersion (precipitation) polymerization and the like are preferable, and solution polymerization and dispersion (precipitation) polymerization are more preferable.

As described above, the block polymer (A) has a mass of the polyethylene oxide portion (A2) / [total mass of the acrylic polymer portion (A1) and the polyethylene oxide portion (A2)] = 0.01 to 0.5. It is preferable that (A2) / [(A1) + (A2)] = 0.05 to 0.3.
The method for obtaining the mass of the polyethylene oxide portion (A2) / [total mass of the acrylic polymer portion (A1) and the polyethylene oxide portion (A2)] in the present invention will be described.
Strictly speaking, when calculating the mass of the polyethylene oxide part (A2), it should be determined by calculating only the weight of the polyethylene oxide in the azo polymerization initiator. In the present invention, the polyethylene oxide part (A2) is calculated. The weight of the polymer azo polymerization initiator having the above is applied as it is. The mass of the acrylic polymer portion (A1) is the total amount of acrylic monomers used for polymerization.
That is, when synthesized using 90 parts by weight of a monomer and 10 parts by weight of a polymer azo polymerization initiator having a polyethylene oxide part (A2), the mass of the polyethylene oxide part (A2) / [acrylic polymer part (A1) and The total mass of the polyethylene oxide part (A2)] is 0.1.

The mass average molecular weight of the block polymer (A) is preferably from 3,000 to 1,000,000, more preferably from 5,000 to 500,000, from the viewpoint of handling properties and application to medical devices and the like. It is.
The mass average molecular weight of the block polymer (A) can be adjusted, for example, by appropriately changing the ratio of the total number of moles of the acrylic monomer to the number of moles of the azo group contained in the polymer azo polymerization initiator. It becomes possible. For example, if the ratio of the total number of moles of acrylic monomers to the number of moles of azo groups contained in the polymer azo polymerization initiator is 200, a 200-mer acrylic polymer is incorporated between PEG chains. Thus, a high cohesive force due to the entanglement of the polymer can be imparted.

The glass transition temperature of the block polymer (A) is preferably −70 ° C. to 50 ° C., more preferably −60 ° C. to 40 ° C. By preparing the glass transition temperature of the block polymer (A) in the range of −70 ° C. to 50 ° C., good embedding property with respect to the medical device body becomes possible, and adhesion to the medical device body can be imparted. Furthermore, the workability at the time of applying to a molded body can be improved.
The glass transition temperature can be adjusted by appropriately preparing the monomer constituting the acrylic polymer part (A1). For example, by increasing the ratio of polyalkoxyalkyl (meth) acrylates such as 2-methoxyethyl (meth) acrylate, flexibility unique to the side chain ether bond can be imparted, so the glass transition temperature is −50 ° C. Can be prepared in a range close to.

<Crosslinking agent>
Next, the crosslinking agent that can be used in the present invention will be described. Examples of the crosslinking agent that can be used in the present invention include those having at least one functional group selected from an epoxy group, an isocyanate group, and an aziridinyl group, a metal chelate compound, a carbodiimide group-containing compound, and the like. These crosslinking agents can be used for the purpose of increasing the elastic modulus and resistance of the coating film, or for adjusting the adhesive force.

[Crosslinking agent having epoxy group]
The crosslinking agent having an epoxy group used in the present invention is not particularly limited as long as it has two or more epoxy groups in one molecule.
Examples of the crosslinking agent having a bifunctional epoxy group include ethylene glycol diglycidyl ether, polyethylene oxide diglycidyl ether, propylene glycol diglycidyl ether, polypropylene glycol diglycidyl ether, tetramethylene glycol diglycidyl ether, and polytetramethylene glycol diglycidyl. Ether, 1,6-hexanediol diglycidyl ether, neopentyl glycol diglycidyl ether, aliphatic epoxy compounds such as polybutadiene diglycidyl ether, bisphenol A type epoxy resin, bisphenol F type epoxy resin, bisphenol S type epoxy resin, biphenol type Epoxy resin, dihydroxybenzophenone diglycidyl ether, resorcinol diglycidyl ether Ter, hydroquinone diglycidyl ether, dihydroxyanthracene type epoxy resin, bisphenol fluorenediglycidyl ether, N, N-diglycidyl aniline and other aromatic epoxy compounds, hydrogenated aromatic epoxy compounds as described above, hexahydrophthalic acid di Examples include alicyclic epoxy compounds such as glycidyl esters.
Examples of the crosslinking agent having three or more epoxy groups include triglycidyl isocyanurate, trisphenol type epoxy compound, tetrakisphenol type epoxy compound, phenol novolac type epoxy compound and the like.

[Crosslinking agent having isocyanate group]
The crosslinking agent having an isocyanate group used in the present invention is not particularly limited as long as it is a compound having two or more isocyanate groups in one molecule.
Examples of the bifunctional isocyanate compound include 2,2′-diphenylmethane diisocyanate, 2,4′-diphenylmethane diisocyanate, 4,4′-diphenylmethane diisocyanate, 2,4-tolylene diisocyanate, 2,6-tolylene diisocyanate, hexa Examples thereof include methylene diisocyanate and isophorone diisocyanate.
Examples of the trifunctional isocyanate compound include a trimethylolpropane adduct of diisocyanate described above, a burette reacted with water, and a trimer having an isocyanurate ring.
Moreover, the isocyanate group in the crosslinking agent which has an isocyanate group may be blocked, and does not need to be blocked.
The blocked isocyanate crosslinking agent used in the present invention may be a blocked isocyanate compound in which the isocyanate group in the isocyanate compound is blocked with ε-caprolactam, MEK oxime, cyclohexanone oxime, pyrazole, phenol, etc., and is particularly limited. Is not to be done.

[Crosslinking agent having aziridinyl group]
The aziridine compound used in the present invention is not particularly limited as long as it is a compound having two or more aziridine groups in one molecule. Examples of the aziridine compound include 2,2′-bishydroxymethylbutanol tris [3- (1-aziridinyl) propionate], 4,4-bis (ethyleneiminocarbonylamino) diphenylmethane, and the like.

[Metal chelate compounds]
Examples of the metal chelate compound used in the present invention include organoaluminum compounds such as aluminum chelate compounds, aluminum alkoxide compounds and aluminum acylate compounds, organotitanium compounds such as titanium chelate compounds, titanium alkoxide compounds and titanium acylate compounds, zirconium Examples thereof include organic zirconium compounds such as chelate compounds, zirconium alkoxide compounds, and zirconium acylate compounds.

[Carbodiimide group-containing compound]
As the carbodiimide group-containing compound used in the present invention, carbodilite series of Nisshinbo Co., Ltd. can be used, and water-based types such as V-02, V-04 and V-06, V-01 and V-03. , V-05, V-07, V-09 and other oily types.

[Β-Hydroxyalkylamide group-containing compound]
In the present invention, a β-hydroxyalkylamide group-containing compound can also be used as a crosslinking agent.
The β-hydroxyalkylamide group-containing compound is not particularly limited as long as it is a compound containing a β-hydroxyalkylamide group in the molecule. Examples of the β-hydroxyalkylamide group-containing compound include various compounds including N, N, N ′, N′-tetrakis (hydroxyethyl) adipamide (PrimidXL-552 manufactured by Ems Chemie).

  In the present invention, the crosslinking agent may be used alone or in combination. The amount of the crosslinking agent used may be determined in consideration of the type and mole number of functional groups contained in the block polymer (A), and is not particularly limited, but is usually a block polymer (A). It is used in the range of 0.1 to 10 parts by mass with respect to 100 parts by mass. By blending in a range smaller than the number of moles of the functional group contained in the block polymer (A), the concern that the unreacted crosslinking agent is liberated can be eliminated. Within this range, particularly excellent performance is exhibited in each of the effects of intended blood compatibility, biological tissue compatibility, biological tissue adhesion suppression, plasma protein adsorption, hemolysis, and cytotoxicity.

<Use as biocompatible material>
Since the block polymer (A) has low plasma protein adsorption and platelet adhesion, all disposable medicines used in combination with anticoagulants such as vacuum blood collection tubes, blood bags, extracorporeal circuits, etc. It can be suitably used for medical devices that come into contact with blood, such as devices, medical devices such as heart-lung machines and stents, and artificial organs.
Moreover, the coating agent containing a block polymer (A) is excellent in coating property. Therefore, a blood contact medical device (2) can be provided by applying a coating agent on the surface of the blood contact medical device (1) and then drying or curing to form a blood contact coating film. .

  In addition, since the block polymer (A) has low cytotoxicity and low plasma protein adsorptivity, the block polymer (A) is applied to the surface of the cell culture member (3), and then dried or cured to form a cell culture coating film. The cell culture member (4) which is formed and is one of research materials for drug discovery research and regenerative medicine research can be provided. The coating film for cell culture is not a medium for cells to be cultured, but supports and promotes rapid growth and proliferation of cells.

As a method for applying to the medical device (1) or the cell culture member (3), the solution is applied to the substrate by various application methods such as brush, dipping, roller, spray, injection, and coating machine, and dried. Is done. On the other hand, examples of the coating method for the film include dip coating, comma coating, gravure coating, curtain coating, die coating, lip coating, micro gravure coating, slot die coating, reverse coating, and kiss coating. After coating, the solvent is removed, and the film thickness of the coated film is not limited, but is preferably 1 mm or less.
Examples of the solvent include water, methanol, ethanol, acetone, organic solvents such as methyl ethyl ketone (hereinafter referred to as MEK), and the like.

  Examples of the material for the medical device (1) and the cell culture member (3) include polyester, polystyrene, silicone, polyamide, polyimide, polyolefin, polysulfone, polyethersulfone, polyethylene, polypropylene, polyvinyl chloride, and polyvinylidene chloride. , Resins such as polyvinylidene fluoride, polytetrafluoroethylene, halogenated polyolefin, polycarbonate, polyvinylidene fluoride, and the like. In addition to the resin, metal materials such as stainless steel, titanium, and titanium alloys, ceramics such as hydroxyapatite, graphite, and titanium nitride can be used.

The form of the medical device (1) and the cell culture member (3) may be a three-dimensional molded body, a sheet shape, or a thread shape.
Examples of the sheet-like material include a film, a nonwoven fabric, and paper.
Examples of the film include various plastic films such as silicone, PET, cellulose ester, polyethylene, polypropylene, ethylene / vinyl acetate copolymer, polyvinyl chloride, and polyurethane. As the cell culture member (3), it is preferable to use a silicone film or a nonwoven fabric from the viewpoint of oxygen permeability.

  The polymer (A) of the present invention can be stably present at the interface in contact with blood, biological tissue, protein, and cells by being applied to the above-mentioned substrate, molded body, and film. There is very little risk of polymer elution. As a result of holding the copolymer at the interface, effects such as inhibition of protein and cell adsorption, inhibition of platelet adhesion and activation are exhibited.

  Furthermore, the polymer (A) of the present invention can be used not only by applying it to the medical device (1) or the cell culture member (3) but also making the polymer (A) into a sheet form or a thread form. . For example, it is also possible to obtain a structural material such as a hollow fiber for dialysis or artificial heart lung from the polymer (A) of the present invention.

  The following examples further illustrate the present invention, but the following examples do not limit the scope of the present invention in any way. In the examples, “parts” and “%” represent “parts by weight” and “% by weight”, respectively, Mw means mass average molecular weight, and Tg means glass transition temperature.

<Measurement method of mass average molecular weight (Mw)>
Mw was measured using Showa Denko GPC (Gel Permeation Chromatography) “GPC-101”. GPC is liquid chromatography that separates and quantifies substances dissolved in a solvent (THF; tetrahydrofuran) based on the difference in molecular size. For the measurement in the present invention, “KF-805L” (manufactured by Showa Denko KK: GPC column: 8 mm ID × 300 mm size) is connected in series to the column, the sample concentration is 1 wt%, the flow rate is 1.0 ml / min, the pressure The measurement was performed under the conditions of 3.8 MPa and a column temperature of 40 ° C., and the mass average molecular weight (Mw) was determined in terms of polystyrene. In the data analysis, a calibration curve, molecular weight, and peak area were calculated using software built in the manufacturer, and a mass average molecular weight was obtained with a retention time of 15 to 30 minutes as an analysis target.

<Measuring method of glass transition temperature (Tg)>
About the block copolymer from which the solvent has been removed by drying, “DSC-1” manufactured by METTLER TOLEDO is used, a sample amount of about 5 mg is weighed in an aluminum standard container, the temperature modulation amplitude is ± 1 ° C., and the temperature modulation cycle is 60 seconds. The glass transition temperature was measured from −80 to 200 ° C. under a temperature rising rate of 2 ° C./min, and the glass transition temperature was determined from the differential heat curve of the reversible component.

<Synthesis of block copolymer>
[Production Example 1]
In a reaction vessel equipped with a thermometer, a stirrer, a nitrogen introducing tube, a reflux condenser, and a dropping tube, 65 parts by weight of MEK as an organic solvent was charged in a nitrogen stream and heated at 80 ° C. for 30 minutes with stirring. In the dropping tube, 49.4 parts by weight of methoxyethyl acrylate as a monomer, 0.6 parts by weight of acrylic acid, 2.5 parts by weight of VPE0201 (manufactured by Wako Pure Chemical Industries, Ltd .: Macroazo Initiator) as a polymerization initiator, and as a solvent MEK was charged in 5 parts by weight and added dropwise over 2 hours. After completion of the dropwise addition, the mixture was aged for 1 hour, and then a solution in which 0.0125 parts by weight of 2,2′-azobis (isobutyric acid) dimethyl was dissolved in 2 parts by weight of MEK was added and aged for 1 hour. Thereafter, a solution in which 0.0125 parts by weight of 2,2′-azobis (isobutyric acid) dimethyl was dissolved in 2 parts by weight of MEK was further added, aged for 1 hour, and then cooled to room temperature to stop the reaction. Then, after removing MEK with a diaphragm pump, ethanol was added and diluted to obtain a block polymer solution having a solid content of 10% by weight. Mw of the obtained block copolymer was 281,000, Tg was -45 ° C, and the proportion of PEG was 4.8% by weight.

[Production Examples 2 to 6]
Synthesis was performed in the same manner as in Production Example 1 according to the composition and parts by weight of Table 1, and a block copolymer solution was obtained. The characteristic values are shown in Table 1.

[Production Example 7]
A reaction vessel equipped with a thermometer, a stirrer, a nitrogen inlet tube, and a reflux condenser was charged with 526 parts by weight of ion-exchanged water as a solvent, 29 parts by weight of methanol, and 50 parts by weight of methoxyethyl acrylate as a monomer under a nitrogen stream. The bottom was heated to 80 ° C. Separately, as a polymerization initiator, a solution prepared by dissolving 8.8 parts by weight of VPE0201 (manufactured by Wako Pure Chemical Industries, Ltd .: Macroazo Initiator) with 20 parts by weight of ion-exchanged water was prepared, and 30 times after the solvent in the reaction vessel was refluxed. Added after a minute. After completion of the addition, the mixture was aged for 5 hours, and then cooled to stop the reaction. After removing methanol with a diaphragm pump, ion exchange water was added to dilute to obtain a block polymer aqueous dispersion having a solid content of 10% by weight. The obtained block copolymer had a Tg of −50 ° C. and a PEG ratio of 15.0% by weight. Mw is not measured because the resin is insoluble in THF.

[Production Example 8]
In the same manner as in Production Example 7, synthesis was carried out according to the composition and parts by weight of Table 1 to obtain a block copolymer solution. The characteristic values are shown in Table 1.

[Comparative Production Example 1]
In a reaction vessel equipped with a thermometer, a stirrer, a nitrogen introducing tube, a reflux condenser, and a dropping tube, 65 parts by weight of MEK as an organic solvent was charged in a nitrogen stream and heated at 80 ° C. for 30 minutes with stirring. In a dropping tube, 50 parts by weight of ethyl acrylate as a monomer, 0.3 part by weight of 2,2′-azobis (isobutyric acid) dimethyl as a polymerization initiator, and 5 parts by weight of MEK as a solvent were added dropwise over 2 hours. After completion of the dropwise addition, the mixture was aged for 1 hour, and then a solution in which 0.0125 parts by weight of 2,2′-azobis (isobutyric acid) dimethyl was dissolved in 2 parts by weight of MEK was added and aged for 1 hour. Thereafter, a solution in which 0.0125 parts by weight of 2,2′-azobis (isobutyric acid) dimethyl was dissolved in 2 parts by weight of MEK was further added, aged for 1 hour, and then cooled to room temperature to stop the reaction. Then, after removing MEK with a diaphragm pump, an acrylic resin solution having a solid content of 10% by weight was obtained by adding ethanol and diluting. Mw of the obtained acrylic resin was 62,000, Tg was −18 ° C., and the proportion of PEG was 0% by weight.

[Comparative Production Examples 2-3]
Synthesis was performed in the same manner as in Production Example 1 according to the composition and parts by weight of Table 1, and a block copolymer solution was obtained. The characteristic values are shown in Table 1.

[Comparative Production Example 4]
A reaction vessel equipped with a thermometer, a stirrer, a nitrogen inlet tube, and a reflux condenser was charged with 50 parts by weight of toluene and 36.2 parts by weight of polyethylene glycol having a hydroxyl value of 279 mgKOH / g under a nitrogen stream and up to 110 ° C. with stirring. The temperature was raised, 13 parts by weight of toluene was distilled off, and at the same time, the raw material was dehydrated. After cooling to 50 ° C., 19.1 parts by weight of isophorone diisocyanate was added, the temperature was raised to 110 ° C., and the reaction was continued until the peak derived from the isocyanate group (near 2254 cm −1) disappeared by FT-IR. After stopping the reaction, toluene was removed with a diaphragm pump, and ethanol was added for dilution to obtain a PEG-containing urethane solution having a solid content of 10% by weight. Mw of the obtained resin was 43,000, Tg was −28 ° C., and the ratio of PEG was 65.5% by weight.

[Comparative Production Example 5]
In a reaction vessel equipped with a thermometer, a stirrer, a nitrogen inlet tube, and a reflux condenser, 50 parts by weight of toluene, 43.0 parts by weight of polyethylene glycol having a hydroxyl value of 111 mgKOH / g, and dimethylolbutanoic acid 1.1 were added under a nitrogen stream. The temperature was raised to 110 ° C. with stirring, 14 parts by weight of toluene was distilled off, and the raw material was dehydrated at the same time. After cooling to 50 ° C., 10.6 parts by weight of isophorone diisocyanate was added, the temperature was raised to 110 ° C., and the reaction was carried out until the peak derived from the isocyanate group (near 2254 cm −1) disappeared by FT-IR. After stopping the reaction, toluene was removed with a diaphragm pump, and ethanol was added for dilution to obtain a PEG-containing urethane solution having a solid content of 10% by weight. Mw of the obtained resin was 38,000, Tg was −51 ° C., the ratio of PEG was 78.6% by weight, and the acid value was 7.7 mgKOH / g.

In Table 1, abbreviations are as follows.
MEA: Methoxyethyl acrylate, Log P = 0.48
AA: Acrylic acid, Log P = 0.38
THFA: tetrahydrofurfuryl acrylate, Log P = 0.78
HEA: 2-hydroxyethyl acrylate, LogP = 0.12
BA: Butyl acrylate, Log P = 1.88
HEAA: N-hydroxyethylacrylamide, LogP = −0.56
130A: Methoxy-polyethylene glycol acrylate, Log P = −0.76
POA: Phenoxyethyl acrylate, LogP = 2.3
EA: ethyl acrylate, Log P = 0.98
ACMO: acryloylmorpholine, LogP = −0.2
2EHA: 2-ethylhexyl acrylate, LogP = 3.53
V601: 2,2'-azobis (isobutyric acid) dimethyl

[Examples 1, 6 to 10], [Comparative Examples 1 to 5]
Ethanol was added to each of the block polymers described in Production Examples 1 to 6 and Comparative Production Examples 1 to 5 so that the solid content was 5% to obtain a coating agent. Hereinafter, solubility in water, cytotoxicity, spheroid formation, protein adsorption, hemolysis, and coating processability were evaluated. The results are shown in Table 2.

[Examples 11 and 12]
Ion exchange water was added to each of the block copolymers described in Production Examples 7 and 8, and the solid content was adjusted to 5% to obtain a coating agent. The evaluation was made in the same manner as in Example 1.

[Examples 13 and 14]
Each block copolymer described in Production Example 8 and a crosslinking agent were blended at the solid content ratio shown in Table 2, and the solid content was adjusted to 5% with ion-exchanged water to obtain a coating agent. The evaluation was made in the same manner as in Example 1.

[Examples 2 to 5], [Comparative Examples 6 to 7]
In the solid content ratio shown in Table 2, each block polymer described in Production Example 1 and Comparative Production Example 5 and a crosslinking agent are blended, and the solid content is adjusted to 5% with ethanol to obtain a coating agent. It was. The evaluation was made in the same manner as in Example 1.

In Table 2, abbreviations are as follows.
ALCH: Kawaken Fine Chemical Co., Aluminum Chelate Compound Chemitite PZ: Nippon Shokubai Co., Ltd., Polyfunctional Aziridine Compound EX321L: Nagase ChemteX Corp., Multifunctional Aliphatic Epoxy Compound V05: Nisshinbo Co., Ltd., Oily Carbodiimide Group-Containing Compound V02: Nisshinbo Co., Ltd., aqueous carbodiimide group-containing compound XL552: manufactured by Ms Chemie, hydroxyalkylamide group-containing compound

<Evaluation>
(1) Solubility in water About 2 g of the coating agent prepared in Examples and Comparative Examples was added to a precisely weighed shallow metal container and dried at 150 ° C. for 10 minutes. After taking out from the oven and accurately weighing the entire shallow metal container, 5 g of ion-exchanged water was added to the shallow metal container and allowed to stand overnight. After ion-exchanged water was sucked and discharged from the shallow metal container, it was again dried at 150 ° C. for 10 minutes to accurately measure the shallow metal container. The solubility in water was calculated by the following formula, and the solubility in water was evaluated based on three-stage evaluation criteria.
Solubility in water = 100 − [(z−x) / (y−x)] × 100
x: mass of shallow metal container y: mass before treatment with ion-exchanged water z: mass after treatment with ion-exchanged water a: solubility in water ≦ 5%
b: 5% <solubility in water ≦ 50%
c: 50% <solubility in water

(2) Cytotoxicity About 0.5 ml of the coating agent prepared in Examples and Comparative Examples was injected into each well into a U-shaped 96-well plate. This was sucked out and then dried at 50 ° C. for 3 hours to prepare a plate whose inner surface was coated with a coating agent.
After sterilizing this with ethylene oxide gas, 10% fetal bovine serum (FBS) was added to Dulbecco's modified Eagle's medium (DMEM), and the mouse fibroblast cell line (NIH / 3T3 cells) was added per well. 1 × 10 4 cells were seeded and cultured in an incubator with 5% CO 2/37 ° C. until the fifth day.
Cytotoxicity was assessed by performing an ATP assay after 1, 2, 5 days. Specifically, 100 μl of ATP reagent (“Clot” ATP measurement reagent: manufactured by Toyo B-Net) was added to the well after culture, pipetted 5 times, allowed to stand at room temperature for 5 minutes, and then 100 ml of reagent. -The cell lysate was dispensed to another plate and stirred for 1 minute. The amount of luminescence was measured using Mitras LB940 (manufactured by Berthold).
Cytotoxicity = (Luminescence after 5 days in culture) / (Luminescence after 1 day in culture) × 100
a: 80% ≦ cytotoxic b: 20% ≦ cytotoxic <80%
c: cytotoxicity <20%

(3) Spheroid formation For spheroid formation, a well plate prepared by the same method as in (2) above was used, and the cell culture state after 5 days was photographed with a transmission optical microscope 40 times to observe the cell morphology. Was evaluated by
a: one spheroid formed b: a plurality of spheroids formed c: no spheroid formed

(4) Protein adsorptivity About 5 mL of the coating agent prepared in Examples and Comparative Examples was poured into a polystyrene test tube (12 × 75 mm), allowed to stand, then sucked out and dried at 50 ° C. for 3 hours. A test tube whose inner surface was coated with a coating agent was obtained.
Protein-containing solution in which bovine serum albumin is added to phosphate buffer (PBS) to a concentration of 0.5% by weight: 4.0 mL is placed in the test tube, incubated for 24 hours, and then the solution in the test tube is discarded. PBS: Washed with about 4.0 mL, and the washing solution was discarded. Thereafter, the protein adsorbed solution in which 1% by weight of sodium dodecyl sulfate is added to PBS: 4.0 mL is added to elute the protein adsorbed on the inner wall of the test tube, and the protein concentration in the protein eluting solution is determined. It was measured by MicroBCA Protein Assay Kit (manufactured by Takara Bio Inc.), and the amount of protein adsorbed on the coating film was determined.
a: Protein adsorption amount ≦ 0.60 μg
b: 0.60 μg <protein adsorption amount ≦ 2.00 μg
c: 2.00 μg <protein adsorption amount

(5) Hemolysis test The coating agent prepared in Examples and Comparative Examples was applied to a polyethylene terephthalate (PET) film so that the film thickness after drying was 2 to 3 μm, and dried at 100 ° C. for 2 minutes. Aged at 50 ° C. for 3 hours.
10 mL of physiological saline was added to the test piece cut out to 60 cm ² and extracted at 37 ° C. for 72 hours to obtain 10 mL of a test solution.
0.2 mL of rabbit defibrinated blood was added to this test solution and incubated at 37 ° C. for 1, 2 and 4 hours. The absorbance at 576 nm was measured for the supernatant after centrifuging the test solution after incubation at about 750 × g for 5 minutes. Absorbance was measured in the same manner using 10 mL of physiological saline as a negative control and 10 mL of distilled water added with 0.2 mL of rabbit defibrinated blood. The hemolysis rate was calculated by the following formula, and hemolysis (hemolysis toxicity) was evaluated.
Hemolysis rate (%) = (absorbance of specimen−absorbance of negative control) / (absorbance of positive control−absorbance of negative control) × 100
a: hemolysis rate ≦ 2
b: 2 <hemolysis rate ≦ 20
c: 20 <hemolysis rate

(6) Coating film processability A coating agent prepared in Examples and Comparative Examples was applied to a polyethylene terephthalate (PET) film so that the film thickness after drying was 20 to 30 μm, and dried at 100 ° C. for 2 minutes. Then, it aged at 50 degreeC for 3 hours. When cutting this coating film, the workability was evaluated based on whether the coating film adhered to the blade. The processability of the coating film was evaluated based on the following three evaluation criteria.
a: The coating film does not adhere to the blade at all b: The coating film slightly adheres to the blade c: The coating film adheres badly to the blade, causing a practical problem

As can be seen from the examples and comparative examples shown in Table 2, the block polymer (A) having the acrylic polymer part (A1) and the polyethylene oxide part (A2) used in Examples 1 to 14, For the polymer having a polyethylene oxide part used in Comparative Examples 2 to 7, since the main chain has an acrylic polymer part composed of a monomer having an average value of LogP of 0 to 2, Solubility was suppressed, and physical properties satisfying all of cytotoxicity, protein adsorption and hemolysis were obtained. On the other hand, even if the main chain has an acrylic polymer portion composed of a monomer having an average value of LogP of 0 to 2, protein adsorption is remarkable if the polyethylene oxide portion (A2) is not present.
Further, by using a cross-linking agent in combination, it was possible to impart better coating film processability as a coating film.

The polymer (A) of the present invention can be used for blood, a part that comes into contact with living tissue, and a part that comes into contact with cells and proteins. Further, it can be developed not only for use on a molded body but also in the form of a film or a sheet, and further applied to structural materials such as dialysis and heart-lung lung fibers.

Claims (5)

A cell culture member (4) comprising a cell culture member (3) and a cell culture coating film located on the surface of the cell culture member (3),
The cell culture coating film,
An acrylic polymer portion (A1) formed from an acrylic monomer having an average value of water / 1-octanol partition coefficient (LogP) of 0 or more and 2 or less, and a polyethylene oxide portion (A2) are represented by the following formula (I And a block polymer (A) bonded with
Cell culture member (4).
  The mass of polyethylene oxide part (A2) contained in the block polymer (A) / [total mass of acrylic polymer part (A1) and polyethylene oxide part (A2)] = 0.01 to 0.5. The member for cell culture (4) according to claim 1. The block polymer (A) has a polyethylene oxide part (A2) represented by the following general formula (II), and uses a polymer azo polymerization initiator having a mass average molecular weight of 5,000 to 100,000. The cell culture member (4) according to claim 1 or 2, which is a block copolymer obtained by polymerizing an acrylic monomer.


In the formula, m and n represent an integer of 1 or more.   The acrylic polymer portion (A1) is a homopolymer portion or a copolymer portion formed only from an acrylic monomer having a water / 1-octanol partition coefficient (LogP) of 0 or more and 2 or less. The member for cell culture (4) according to item 1. On the surface of the cell culture member (3),
An acrylic polymer portion (A1) formed from an acrylic monomer having an average value of water / 1-octanol partition coefficient (LogP) of 0 or more and 2 or less, and a polyethylene oxide portion (A2) are represented by the following formula (I A coating agent containing the block polymer (A) formed by bonding with a), and then dried or cured to provide a cell culture coating (4). Method.