WO2020066608A1

WO2020066608A1 - Material non-adhesive to biological substances, composition, and compound

Info

Publication number: WO2020066608A1
Application number: PCT/JP2019/035630
Authority: WO
Inventors: 陽介山本; 悠太滋野井
Original assignee: 富士フイルム株式会社
Priority date: 2018-09-27
Filing date: 2019-09-11
Publication date: 2020-04-02
Also published as: JPWO2020066608A1; JP7086202B2

Abstract

The present invention provides: a material which is not adhesive to biological substances and is excellent in both non-adhesive properties to cells and adhesive properties to substrates; a composition; and a compound. This material non-adhesive to biological substances contains a polymer containing repeating units derived from a monomer represented by formula (1).

Description

Biomaterial non-adhesive materials, compositions and compounds

The present invention relates to non-adhesive materials, compositions and compounds for biological materials.

Many medical devices such as catheters and syringes generally have various coatings (coatings) on their outer surfaces. By applying such a coating, effects such as easy insertion of the medical device into the body or drainage of bodily fluids from the body can be obtained.

For example, Patent Document 1 discloses a hydrophilic coating composition containing a polyfunctional polymerizable compound having a large number of acrylamide-based functional groups, and discloses that a strong and uniform coating having good abrasion resistance can be provided. Has been stated. In the examples section of Patent Document 1, a polyfunctional polymerizable compound (PEG (polyethylene @ glycol) 1500 diacrylamide) is specifically used.

JP 2011-513566 A

On the other hand, in a coating film (coating) applied to various medical instruments as described above, various properties are required in addition to the abrasion resistance described in Patent Document 1.
For example, non-adhesiveness of cells and substrate adhesion are required.

However, the abrasion-resistant coating described in Patent Literature 1 showed room for improvement in both the non-adhesiveness of cells and the adhesion to substrates, as shown in Comparative Example 5 described below.

Therefore, an object of the present invention is to provide a biological material non-adhesive material which is excellent in both non-adhesiveness of cells and substrate adhesiveness.
Another object of the present invention is to provide a composition and a compound.

The present inventors have conducted intensive studies in order to solve the above-mentioned problems, and as a result, a biomaterial non-adhesive material containing a polymer containing a repeating unit derived from a monomer represented by the formula (1) described below has a non-cellular property. The inventors have found that both the adhesion and the substrate adhesion are excellent, and have completed the present invention.

That is, the present invention includes the following [1] to [10].
[1] A biological substance non-adhesive material containing a polymer containing a repeating unit derived from a monomer represented by the following formula (1).
[2] The biological material non-adhesive material according to [1], wherein R ³³ is any one selected from the group consisting of an ethylene group, a propylene group, a trimethylene group, a propylidene group, and an isopropylidene group. .
[3] The biological substance non-adhesive material according to the above [1] or [2], wherein the monomer represented by the above formula (1) is a monomer represented by the following formula (2).
[4] The non-adhesive biological material according to any one of [1] to [3], wherein R ³¹ is a hydrogen atom.
[5] The non-adhesive biological material according to any one of [1] to [4], wherein R ³² is a trimethylene group.
[6] The biological substance non-adhesive material according to any one of [1] to [5], wherein m is 3 or 4.
[7] The biological substance non-adhesive according to any one of [1] to [6], wherein the biological substance in the biological substance non-adhesive material is at least one selected from the group consisting of cells and proteins. Material.
[8] The biological substance non-adhesive material according to any one of the above [1] to [7], which is used for coating a medical device or coating a cell culture container.
[9] A composition for forming a biological substance non-adhesive material, comprising a monomer represented by the formula (1) and a solvent.
[10] A compound represented by the formula (3).

ADVANTAGE OF THE INVENTION According to this invention, the biological substance non-adhesive material excellent in both the non-adhesion of a cell and a base material adhesion can be provided.
Further, according to the present invention, a composition and a compound can be provided.

範囲 In this specification, the range expressed using “to” includes both ends of “to”. For example, a range represented by “AB” includes A and B.

において In the present specification, the solid content means a component contained in the composition excluding the solvent component, and is calculated as a solid content even if the property is liquid.

As used herein, the term “biological substance” broadly encompasses substances constituting the living body and substances involved in the living body. For example, it is meant to include proteins, cells, tissues where cells are collected, peptides, vitamins, hormones, blood cells, antigens, antibodies, bacteria, viruses, and the like.
Further, in the present specification, the term "non-adhesiveness of biological material" means not only no adhesion at all but also an improvement in adhesion (decrease in the amount of adhesion) before and after application even if adhesion is observed. Means that. Therefore, not only prevention of adhesion but also suppression of adhesion is included as a concept.

A description will be given of an estimating mechanism by which the present invention produces an effect.
The adhesion of the biological material is performed via the protein in the biological tissue fluid adsorbed on the member surface. It is also known that the structure of a protein is changed by adsorption, and cell adhesion is made possible by exposing a cell adhesion site that can be recognized as a scaffold by various cells. The reason why the characteristics of the present invention can be obtained is that the structure derived from the compound represented by the formula (1) in the cured film is hydrophilic, thereby suppressing protein adsorption on the surface. thinking. Further, polyfunctional monomers generally cause curing shrinkage and are liable to cracks, and thus are often not suitable for coating structures having bent portions such as medical instruments and cell culture vessels. It is considered that the film was moderately flexible and the generation of cracks could be suppressed due to the alkyleneoxy group having an appropriate length derived from the compound represented by 1). Therefore, it is considered that the uniformity of the film was increased, water intrusion could be suppressed, and the substrate adhesion (water resistance) was improved.

[Compound]
The present invention provides a compound represented by the formula (3).

In the formula (3), the meaning of each symbol is as follows.
R ³¹ represents a hydrogen atom or a methyl group, and a plurality of R ³¹ may be the same or different. R ³¹ is preferably a hydrogen atom.
m represents an integer of 3 to 5, and is preferably 3 or 4.

具体 Specific examples of the compound represented by the formula (3) are shown below, but it should not be construed that the invention is limited thereto.

<Method of synthesizing compound represented by formula (3)>
The method for synthesizing the compound represented by the formula (3) is not particularly limited. For example, the compound can be synthesized according to the following Scheme 1 or Scheme 2.

In Scheme 1 and Scheme 2, m represents an integer of 3 to 5, and R ³¹ represents a hydrogen atom or a methyl group.

[Composition]
Further, the present invention provides a composition for forming a non-adhesive material for a biological substance, comprising a monomer represented by the formula (1) and a solvent (hereinafter, may be referred to as “the composition of the present invention”). I will provide a. The biological substance non-adhesive material will be described later.

<Monomer represented by Formula (1)>
The monomer represented by the formula (1) contained in the composition of the present invention will be described.

In the formula (1), the meaning of each symbol is as follows.
R ³¹ represents a hydrogen atom or a methyl group, and a plurality of R ³¹ may be the same or different. R ³¹ is preferably a hydrogen atom.

R ³² represents a linear or branched alkylene group, a plurality of R ³² may be different may be the same as each other. R ³² is preferably any one selected from the group consisting of a methylene group, an ethylene group, a propylene group, a trimethylene group, a propylidene group, and an isopropylidene group, and more preferably an ethylene group or a trimethylene group. More preferably, it is a trimethylene group.

R ³³ represents a linear or branched alkylene group, and a plurality of R ³³ may be the same or different. R ³³ is preferably any one selected from the group consisting of an ethylene group, a propylene group, a trimethylene group, a propylidene group, and an isopropylidene group, more preferably an ethylene group or a trimethylene group, More preferably, it is a group.

M represents an integer of 3 to 8, preferably 3 to 5, and more preferably 3 or 4.

モノマー As the monomer represented by the formula (1), a monomer represented by the formula (2) is preferable.

中 In the formula (2), the meaning of each symbol is the same as the same symbol in the formula (1).

具体 Specific examples of the monomer represented by the formula (1) are shown below, but the invention is not limited thereto.

<Content of monomer represented by formula (1)>
In the composition of the present invention, the content of the monomer represented by the formula (1) is not particularly limited, but is preferably 50% by mass or more, more preferably 80% by mass or more in the solid content of the composition of the present invention. Is more preferably 90% by mass or more, and even more preferably 95% by mass or more. The upper limit is not particularly limited, but may be 100% by mass, and is often 99% by mass or less.

<solvent>
The solvent contained in the composition of the present invention will be described.
Although the solvent contained in the composition of the present invention is not particularly limited, for example, water, organic solvents (eg, esters such as ethyl acetate and n-butyl acetate; aromatic hydrocarbons such as toluene and benzene; n-hexane) And aliphatic hydrocarbons such as n-heptane; alicyclic hydrocarbons such as cyclohexane and methylcyclohexane; ketones such as methyl ethyl ketone (MEK), methyl isobutyl ketone and cyclohexanone; alcohols such as methanol and butanol); And a mixed solvent thereof.
Among them, alcohol solvents such as methanol and ethanol are preferred from the viewpoint that surface unevenness during coating is unlikely to occur.

The solvent may be used alone or in combination of two or more.
The content of the solvent in the composition of the present invention (when a plurality of types are present, the total thereof) is not particularly limited, but should be 10% by mass to 95% by mass relative to the total mass of the composition of the present invention. Is preferably 30% by mass to 90% by mass, and more preferably 50% by mass to 80% by mass.

<Components other than monomer and solvent represented by formula (1)>
The composition of the present invention may contain components other than the monomer represented by the formula (1) and the solvent.
Such components include, but are not limited to, monomers other than the monomer represented by formula (1), a polymerization initiator, and other additives.

<< Monomers other than the monomer represented by Formula (1) >>
The composition of the present invention may further contain a monomer other than the monomer represented by the formula (1) in addition to the monomer represented by the formula (1).

The monomer other than the monomer represented by the formula (1) is not particularly limited, and includes at least one selected from the group consisting of commercially available monofunctional monomers, polyfunctional monomers, and betaine monomers described below. Preferably, at least one selected from the group consisting of

When the composition of the present invention contains a monomer other than the monomer represented by the formula (1), the content of the monomer other than the monomer represented by the formula (1) is not particularly limited. Is preferably 50% by mass or less, more preferably 20% by mass or less, still more preferably 10% by mass or less, and even more preferably 5% by mass or less. The lower limit is not particularly limited, but may be 0% by mass.

(Copolymerizable monomer)
The composition of the present invention contains a monomer copolymerizable with the monomer represented by the formula (1) (hereinafter, also simply referred to as a “copolymerizable monomer”) in that the substrate adhesion is further improved. May be.
The copolymerizable monomer is not particularly limited, and includes a monomer having an ethylenically unsaturated group. Examples thereof include (meth) acrylate monomers; (meth) acrylamide monomers; (meth) acrylic acid, crotonic acid, and Monomers having a carboxyl group-containing ethylenically unsaturated group such as itaconic acid (carboxylic acid group-containing monomers); and monomers having a betaine structure. The number of ethylenically unsaturated bonds in the monomer having an ethylenically unsaturated group in the molecule is not particularly limited, but is preferably 1 to 8, more preferably 1 to 4, and still more preferably 1 or 2.
As the copolymerizable monomer, a (meth) acrylate-based monomer, a (meth) acrylamide-based monomer, a monomer having a betaine structure, or the like is preferable.

(Meth) acrylate-based monomers Examples of (meth) acrylate-based monomers include methyl (meth) acrylate, ethyl (meth) acrylate, n-butyl (meth) acrylate, 2-ethylhexyl (meth) acrylate, and 2-methoxyethyl (Meth) acrylate, 2-hydroxyethyl (meth) acrylate, 2-hydroxy-1,3-propylenediol diacrylate, 2-hydroxypropyl (meth) acrylate, 3- (meth) acryloyloxypropylsulfonic acid, N, N- Dimethylaminoethyl (meth) acrylate, N, N-diethylaminoethyl (meth) acrylate, polyalkylene glycol mono (meth) acrylate, polyalkylene glycol di (meth) acrylate, 2- (meth) acryloyloxy Examples include siethyl methyl sulfoxide, tetraethylene glycol dimethacrylate, urethane dimethacrylate, and trimethylolpropane tri (meth) acrylate.

(Meth) acrylamide-based monomer Examples of the (meth) acrylamide-based monomer include (meth) acrylamide, N-methyl (meth) acrylamide, N-2-hydroxyethyl (meth) acrylamide, and a compound represented by the following formula (A-1) And polyfunctional (meth) acrylamide compounds represented by (A-4).

-Betaine monomer The betaine monomer is not particularly limited, and examples thereof include monomers having a betaine structure such as a sulfobetaine structure, a phosphobetaine structure, and a carboxybetaine structure. The skeleton of the betaine monomer that can be included in the composition of the present invention is not particularly limited, but an acrylate monomer or an acrylamide monomer is preferable. Examples of the betaine monomer include those described in International Publication WO2017 / 018146 and the like.
As the above-mentioned betaine monomer, a compound represented by the following formula (C) is particularly preferable in terms of more excellent biocompatibility.

In the formula (C), R ³⁰ represents a hydrogen atom or an alkyl group.
The number of carbon atoms of the alkyl group represented by R ³⁰ is not particularly limited, but is preferably 1 to 15, more preferably 1 to 10, still more preferably 1 to 6, and particularly preferably 1 to 3. The alkyl group may be linear, branched, or cyclic.
Examples of the alkyl group include a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, a t-butyl group, an n-hexyl group, a cyclopentyl group, and a cyclohexyl group.
The alkyl group may have a substituent.
As R ³⁰ , a hydrogen atom or an alkyl group having 1 to 6 carbon atoms is preferable, and a hydrogen atom or an alkyl group having 1 to 3 carbon atoms is more preferable.
L ³⁰ represents an oxygen atom or NR ^A- .
^RA represents a hydrogen atom or an alkyl group. The alkyl group represented by R ^A has the same meaning as the alkyl group represented by R ³⁰ described above, and the preferred embodiment is also the same. Among them, ^RA is preferably a hydrogen atom.
R ³¹ represents a monovalent group represented by the following formula (I), a monovalent group represented by the following formula (II), or a monovalent group represented by the following formula (III).

In the formula (I), L ³¹ and L ³² each independently represent a divalent linking group.
L ³¹ and L ³² are not particularly limited, and may be an alkylene group having 1 to 10 carbon atoms which may contain a hetero atom (which may be any of linear, branched, and cyclic; Is preferred). The number of carbon atoms in the alkylene group is more preferably 1 to 6, more preferably 1 to 4, and particularly preferably 2 to 4.

R ³² and R ³³ each independently represent an alkyl group.
The number of carbon atoms in the alkyl group represented by R ³² and R ³³ is not particularly limited, but is preferably 1 to 6, more preferably 1 to 3. The alkyl group may be linear, branched, or cyclic.
Examples of the alkyl group include a methyl group, an ethyl group, an n-propyl group, and an i-propyl group.
The alkyl group may have a substituent. The substituent which the alkyl group may have is not particularly limited.
* Represents a bonding position.

In formula (II), L ³³ and L ³⁴ each independently represent a divalent linking group.
The divalent linking groups represented by L ³³ and L ³⁴ have the same meanings as the divalent linking groups represented by L ³¹ and L ³² in the above formula (I), and the preferred embodiments are also the same. .
R ³⁴ ~ ^{R 36} each independently represents an alkyl group.
The alkyl group represented by R ³⁴ to R ³⁶ has the same meaning as the alkyl group represented by R ³² and R ³³ in the above formula (I), and the preferred embodiments are also the same.
* Represents a bonding position.

In the formula (III), L ³⁵ and L ³⁶ each independently represent a divalent linking group.
The divalent linking group represented by L ³⁵ and L ³⁶ has the same meaning as the divalent linking group represented by L ³¹ and L ³² in formula (I) described above, and the preferred embodiments are also the same. .
R ³⁷ and R ³⁸ each independently represent an alkyl group.
The alkyl group represented by R ³⁷ and R ³⁸ has the same meaning as the alkyl group represented by R ³² and R ³³ in the above formula (I), and the preferred embodiments are also the same.
* Represents a bonding position.

As R ³¹ in the formula (C), a group represented by the formula (I) or a group represented by the formula (II) is preferable in terms of more excellent biocompatibility.

The betaine monomer can be synthesized by a known method.

具体 Specific examples of betaine monomers are shown below, but the present invention is not limited thereto.

《Polymerization initiator》
The polymerization initiator (hereinafter sometimes referred to as “initiator”) is not particularly limited, but a thermal polymerization initiator or a photopolymerization initiator is preferable.
Examples of the photopolymerization initiator include, for example, an alkynephenone-based photopolymerization initiator, a methoxyketone-based photopolymerization initiator, an acylphosphine oxide-based photopolymerization initiator, and a hydroxyketone-based photopolymerization initiator (for example, Omnirad (registered trademark)) 184; 1,2-α-hydroxyalkylphenone), aminoketone photopolymerization initiators (eg, 2-methyl-1- [4- (methylthio) phenyl] -2-morpholino-propan-1-one (Omnirad (registered trademark) 907)), an oxime-based photopolymerization initiator, and an oxyphenylacetate-based photopolymerization initiator (Omnirad (registered trademark) 754).
Other initiators include, for example, azo-based polymerization initiators (eg, V-50, V-601), persulfate-based polymerization initiators, persulfate-based polymerization initiators, and redox-based polymerization initiators. No.

One type of polymerization initiator may be used alone, or two or more types may be used in combination.
When the composition of the present invention contains a polymerization initiator, the content of the polymerization initiator in the composition (when a plurality of types are present, the total thereof) is not particularly limited. % To 10% by mass, more preferably 0.5% to 8% by mass, still more preferably 1% to 5% by mass.

《Other additives》
Other additives include, for example, polymerization inhibitors, binder resins, polyfunctional amines, polyfunctional thiols, surfactants, plasticizers, surface lubricants, leveling agents, softeners, antioxidants, antioxidants, light Stabilizers, UV absorbers, inorganic or organic fillers, metal powders and the like.
The binder resin is not particularly limited, for example, acrylic resin, styrene resin, vinyl resin, polyolefin resin, polyester resin, polyurethane resin, polyamide resin, polycarbonate resin, polydiene resin, epoxy resin , A silicone-based resin, a cellulose-based resin, and a chitosan-based resin.

[Biomaterial non-adhesive material]
The biomaterial non-adhesive material of the present invention includes a polymer containing a repeating unit derived from the monomer represented by the formula (1).

<Content of repeating unit derived from monomer represented by formula (1) in polymer>
In the biomaterial non-adhesive material of the present invention, the content of the repeating unit derived from the monomer represented by the formula (1) in the polymer is not particularly limited, but is 50 mass% with respect to all the repeating units in the polymer. % Or more, more preferably 80% by mass or more, even more preferably 90% by mass or more, and even more preferably 95% by mass or more. The upper limit is not particularly limited, but may be 100% by mass.

<Polymer content in biomaterial non-adhesive material>
The content of the polymer containing the repeating unit derived from the monomer represented by the formula (1) in the biomaterial non-adhesive material of the present invention is not particularly limited. Is preferably 50% by mass or more, more preferably 80% by mass or more, still more preferably 90% by mass or more, and even more preferably 95% by mass or more. The upper limit is not particularly limited, but may be 100% by mass.

<Production method of non-adhesive material for biological material>
The method for producing the biological material non-adhesive material of the present invention is not particularly limited.
For example, a method using the above-described composition of the present invention can be mentioned. As an example, when producing a biomaterial non-adhesive material in the form of a film, the composition of the present invention is placed on the surface of a medical device or a cell culture container to form a cured film precursor film on the surface, There is a method of curing this to form a biological material non-adhesive material as a cured film.

<Form of non-adhesive biomaterial>
The form of the non-adhesive material for a biological material of the present invention is not particularly limited, but is often in the form of a film.

The method for disposing the composition of the present invention on the surface of a substrate is not particularly limited, and examples thereof include a method using a bar coater, spin coating, dipping, or painting.
The substrate surface may be subjected to a surface treatment such as a plasma treatment and an ozone treatment before disposing the composition of the present invention. Further, the surface of the medical device may be coated.
The method for producing the cured product (cured film) of the present invention is not particularly limited. For example, the above-described composition of the present invention is applied on a substrate, and then heated or irradiated with light (as light, for example, ultraviolet light). , Visible light, X-rays, etc.).

The material of the substrate is not particularly limited, and examples thereof include a metal material, a ceramic material, and a plastic material.
Examples of the type of the plastic material include, for example, polyethylene terephthalate, polybutylene terephthalate, polyethylene naphthalate, polyethylene, polypropylene, cellophane, diacetyl cellulose, triacetyl cellulose, acetyl cellulose butyrate, polyvinyl chloride, polyvinylidene chloride, polyvinyl alcohol, Ethylene-vinyl acetate copolymer, polystyrene, polycarbonate, polymethylpentene, polysulfone, polyetheretherketone, polyethersulfone, polyetherimide, polyimide, fluororesin, nylon, acrylic resin, polyamide, cycloolefin, nylon, and poly Ether sulfone and the like can be mentioned.
Examples of the type of the metal material include gold, stainless steel, cobalt chromium alloy, amalgam alloy, silver palladium alloy, gold silver palladium alloy, titanium, nickel titanium alloy, and platinum.
Examples of the type of the ceramic material include hydroxyapatite.
The shape of the substrate is not particularly limited, and may be a plate shape or a three-dimensional shape.

The method for applying the composition of the present invention is not particularly limited. For example, dipping, roll coating, kiss roll coating, gravure coating, reverse coating, roll brushing, spray coating, dip roll coating, bar coating, spin coating, and knife coating , An air knife coat, a curtain coat, a lip coat, and a method such as an extrusion coat method using a die coater.

The method for heating is not particularly limited, and examples thereof include a method using a blow dryer, an oven, an infrared dryer, a heating drum, and the like.
The heating temperature is not particularly limited, but is preferably from 30 ° C to 150 ° C, more preferably from 40 ° C to 120 ° C.
The heating time is not particularly limited, but is usually 1 minute to 6 hours. When drying in a coating apparatus, the heating time is 1 minute to 20 minutes, and the heating temperature during heating after application (for example, heating in a winding form) is preferably room temperature to 50 ° C.

As a method for irradiating light, for example, a low-pressure mercury lamp, a medium-pressure mercury lamp, a high-pressure mercury lamp, a metal halide lamp, a Deep-UV (ultraviolet) light, an LED (light emitting diode) lamp, a xenon lamp, a chemical lamp, a carbon arc lamp, and the like are used. Method. Although the energy of light irradiation is not particularly ^{limited, 0.1J / cm 2 ~ 10J /} cm 2 is preferred.
The thickness of the cured film is not particularly limited, but is preferably 0.05 μm to 500 μm, more preferably 0.1 μm to 100 μm, and further preferably 1 μm to 50 μm.

[Use]
The use of the non-adhesive material of the present invention is not particularly limited, but it is preferably used for coating medical devices or coating cell culture containers.
The medical device to which the biological material non-adhesive material of the present invention is applied is not particularly limited. For example, artificial blood vessels, hemodialysis membranes, catheters, blood filters, blood storage packs, artificial organs, thrombus collection devices, dentures, and dentures Floors and the like.
The cell culture container to which the biological material non-adhesive material of the present invention is applied is not particularly limited, and examples thereof include a cell culture microplate, a cell culture dish, a cell culture tube, and a cell culture flask.
Further, the biological material in the biological material non-adhesive material is preferably at least one selected from the group consisting of cells and proteins.

Hereinafter, the present invention will be described more specifically with reference to examples, but the present invention is not limited to these examples.

[Synthesis Example 1]
<Synthesis of Monomer (1-1)>
The monomer (1-1) was prepared according to the method of Kimura et al. (Polyacrylamide pseudo-crown ethers via hydrogen bond-assisted cyclopolymerization, Journal of Pharmaceutical Sciences, Journal of Pharmaceutical Science and Technology, Journal of Pharmaceutical Science and Technology). did.

[Synthesis Example 2]
<Synthesis of Monomer (2-1)>
Triethylene glycol (11.4 g, 76.0 mmol), acrylonitrile (20.3 g, 382 mmol), and potassium hydroxide (35 mg, 0.62 mmol) as a catalyst were mixed to obtain a reaction solution. The resulting reaction solution was stirred at 40 ° C. for 24 hours. Ethyl acetate (50 mL) was added to the reaction solution after stirring, and the mixture was purified by silica gel column chromatography (eluent: ethyl acetate only) to obtain an intermediate (M1-1) (19.4 g, yield 99). %).

A solution obtained by diluting the intermediate (M1-1) (18.5 g, 72.2 mmol) in tetrahydrofuran (100 mL) was dropped into about 0.9 mol / L borane-tetrahydrofuran complex (400 mL, 360 mmol) under reflux. did. After the addition was completed, the reaction solution was stirred under reflux for 5 hours. After the reaction solution was cooled to 0 ° C., methanol (125 mL) and concentrated hydrochloric acid (6 mL) were added to quench the reaction. The reaction solution was filtered through celite, and the obtained filtrate was concentrated to dryness to obtain an intermediate (M2-1) (25.9 g). The obtained crude product was used for the next reaction without purification.

Intermediate (M2-1) (25.9 g, theoretical maximum content: 72.2 mmol) of the crude product obtained in the above reaction, acetonitrile (40 mL), water (80 mL), and sodium hydrogen carbonate (29. 1 g, 347 mmol) and 4-hydroxy TEMPO (4-hydroxy-2,2,6,6-tetramethylpiperidine 1-oxyl) (10 mg, 6 μmol) were mixed, and the temperature was lowered to 0 ° C. 3-Chloropropionyl chloride (22.0 g, 172 mmol) was added dropwise to the reaction solution. The reaction solution was heated to 30 ° C. and stirred for 30 minutes.

After allowing the reaction solution to stand, the aqueous layer was removed to obtain an organic layer. The temperature of the organic layer was lowered to 10 ° C., an aqueous sodium hydroxide solution (60 mL) adjusted to 5 mol / L was added, the temperature was raised to 40 ° C., and the mixture was stirred for 2 hours. After allowing the reaction solution to stand, the aqueous layer was removed, and hydrochloric acid was added to the organic layer to adjust the pH to 9 to 10. 4-Hydroxy TEMPO (10 mg, 6 μmol) was added to the obtained organic layer, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained crude product was purified by subjecting it to silica gel column chromatography (eluent: ethyl acetate only to ethyl acetate / methanol = 7: 3) to obtain monomer (2-1) (10.1 g, yield 38%). ) Got. The obtained monomer (2-1) was confirmed to be the target substance by ¹ H NMR (Nuclear Magnetic Resonance).

¹ H NMR (DMSO-d ₆ , 400 MHz) δ: 1.65 (4H, m), 3.16 (4H, q), 3.40 (4H, t), 3.43-3.54 (12H, m), 5.56 (2H, dd), 6.02-6.24 (4H, m), 8.05 (2H, s).

[Synthesis Example 3]
<Synthesis of Monomer (2-2)>
Monomer (2-2) was synthesized in the same manner as monomer (2-1) according to the following route. The obtained monomer (2-2) was confirmed to be the desired product by ¹ H NMR.

¹ H NMR (DMSO-d ₆ , 400 MHz) δ: 1.66 (4H, m), 3.17 (4H, q), 3.41 (4H, t), 3.45-3.53 (16H, m), 5.56 (2H, dd), 6.02-6.24 (4H, m), 8.05 (2H, s).

[Synthesis Example 4]
<Synthesis of Monomer (2-7)>
The intermediate (M2-1) (5 g, 18.9 mmol) obtained in Synthesis Example 1, tetrahydrofuran (60 mL), and methacryloyl chloride (4.35 g, 41.6 mmol) were mixed, and the mixture was heated to 0 ° C. The temperature has dropped. After triethylamine (4.40 g, 43.5 mmol) was added dropwise to the reaction solution, the temperature was raised to room temperature, and the mixture was stirred for 3 hours. After water was added to the reaction solution and extracted with ethyl acetate, the organic layer was dried over magnesium sulfate, filtered, and the filtrate was collected. The filtrate was concentrated under reduced pressure. The obtained crude product was purified by subjecting it to silica gel column chromatography (eluent: ethyl acetate only to ethyl acetate / methanol = 7: 3) to give the monomer (2-7) (3.7 g, yield 49). %). The obtained monomer (2-7) was confirmed to be the desired product by ¹ H NMR.

[Examples 1 to 8, Comparative Examples 1 to 6]
<Preparation of curable composition>
Each component shown in Table 1 was dissolved in a solvent (methanol) to prepare curable compositions (Examples 1 to 8 and Comparative Examples 1 to 6) having a solid content of 20% by mass. In the curable composition, the solid content means all components other than the solvent.

The compounds (monomers) used in the examples and comparative examples are as follows.
Monomer (1-1) Synthesized in Synthesis Example 1 Monomer (2-1) Synthesized in Synthesis Example 2 Monomer (2-2) Synthesized in Synthesis Example 3 Monomer (2-7) Synthesized in Synthesis Example 4 What

{Monomer (A)} Compound represented by formula (A) (manufactured by Tokyo Chemical Industry Co., Ltd.)

{Monomer (B)} Compound represented by formula (B) (synthesis method will be described later)

{Monomer (C)} Compound represented by formula (C) (synthesis method will be described later)

Both the monomer (B) and the monomer (C) were prepared using pentaerylene glycol and hexaethylene glycol as raw materials, and were described in Mabuchi et al. (Improvement of solid material for affinity resin resins by application FK506, Bioorganic & Medicinal Chemistry Letters, 2015, July 15, 25 (14): 2788-2792).

{Monomer (D)} Compound represented by Formula (D)

The monomer (D) is synthesized according to the method described in JP-A-2017-57350.

{Monomer (E)} Compound represented by Formula (E)

The monomer (E) was synthesized according to the method described in paragraph 0080 of JP-T-2011-513566. In addition, n in the said monomer (E) was about 33.

<< MPC >> 2- (methacryloyloxy) ethyl phosphorylcholine (manufactured by Tokyo Chemical Industry Co., Ltd.)

{SB} Sulfobetaine monomer represented by the following formula

Ｓ The SB used in the examples and comparative examples was synthesized with reference to International Publication No. WO2016 / 067995.

The polymerization initiators used in Examples and Comparative Examples are as follows.
Omnirad (registered trademark) 2959 (manufactured by IGM Resins)

<Evaluation of cell adhesion>
The surface of a well of a polystyrene 96-well microplate (manufactured by Evergreen Scientific) was subjected to surface treatment using oxygen plasma (130 V, 120 s) using a plasma processing apparatus (IP-200, manufactured by Izumi Kogyo).
The curable composition prepared above was applied to the surface of a polystyrene 96-well microplate that had been subjected to a surface treatment, and dried to form a cured film precursor film.
Thereafter, the cured film precursor film was exposed to an exposure amount of ² J / cm ² using an ultraviolet exposure machine (ECS-401G, manufactured by Eye Graphic Co., Ltd .; the light source was a high-pressure mercury lamp), cured, and cured. A sample for evaluating adhesion was prepared.
Using the mouse-derived fibroblasts (3T3), cell adhesion was evaluated using the prepared cell adhesion evaluation sample 1.
Using a sample for evaluating cell adhesion, the cells were dispersed in Dulbecco's modified Eagle's medium at a seeding density of 1.0 × 10 ⁴ cells / cm ² , and 37 ° C. in the presence of 5% CO ₂ using a CO ₂ incubator. For 72 hours.
Then, the sample for cell adhesion evaluation was taken out, and it was confirmed using a phase contrast microscope (an inverted research microscope, manufactured by Olympus Corporation) whether or not the cells were attached. The magnification was 4 times.
This operation was performed on 20 of the 96 wells of the sample for cell adhesion evaluation, and the cell adhesion was evaluated according to the following evaluation criteria based on the number of wells to which the cells had adhered.
A: 0 to 1 B: 2 to 3 C: 4 to 6 D: 7 or more Each evaluation result is shown in Table 1.

<Evaluation of substrate adhesion (water resistance)>
The surface of the polystyrene sheet was subjected to a surface treatment using oxygen plasma (130 V, 120 seconds) using a plasma processing apparatus (IP-200, manufactured by Izumi Kogyo KK).
The curable composition prepared above was applied to the surface of the polystyrene sheet subjected to the surface treatment, and dried to form a cured film precursor film.
Thereafter, the cured film precursor film was exposed to an exposure amount of ² J / cm ² using an ultraviolet exposure machine (ECS-401G, manufactured by Eye Graphic Co., Ltd .; the light source was a high-pressure mercury lamp), cured, and cured. A sample for evaluating material adhesion was prepared.
The prepared sample for evaluating substrate adhesion was immersed in PBS (phosphate buffered saline) at 37 ° C. for 72 hours. The substrate adhesion evaluation sample was pulled up from PBS, and the substrate adhesion was evaluated from the area of the cured film remaining on the polystyrene sheet (hereinafter, also referred to as “residual cured film”). The area of the remaining cured film with respect to the area of the polystyrene sheet was expressed as a percentage as a coating rate, and the substrate adhesion was evaluated according to the following evaluation criteria.
A: The coating rate is 95% or more. B: The coating rate is 80% or more and less than 95%. C: The coating rate is less than 80%.

In Examples 1 to 8, the cell adhesion and the substrate adhesion were good.
Above all, Examples 2 and 4 were evaluated as A for both cell adhesion and substrate adhesion, and were particularly excellent.

Claims

A biomaterial non-adhesive material containing a polymer containing a repeating unit derived from the monomer represented by the formula (1).

In the formula (1), R 31 represents a hydrogen atom or a methyl group, a plurality of R 31 may be the same or different, and R 32 is a linear or branched alkylene group. And a plurality of R 32 may be the same or different from each other; R 33 represents a linear or branched alkylene group; and a plurality of R 33 may be the same as each other. M may be different, and m represents an integer of 3 to 8.
Wherein R 33 is an ethylene group, a propylene group, a trimethylene group is any one selected from the group consisting of propylidene, and isopropylidene groups, biological material non-adhesive material according to claim 1.
The biomaterial non-adhesive material according to claim 1 or 2, wherein the monomer represented by the formula (1) is a monomer represented by the formula (2).

The meaning of each symbol in the formula (2) is the same as the same symbol in the formula (1).
The biological substance non-adhesive material according to any one of claims 1 to 3, wherein the R 31 is a hydrogen atom.
Wherein R 32 is a trimethylene group, a biological material non-adhesive material according to any one of claims 1-4.
生体 The biological material non-adhesive material according to any one of claims 1 to 5, wherein m is 3 or 4.
The biological material non-adhesive material according to any one of claims 1 to 6, wherein the biological material in the biological material non-adhesive material is at least one selected from the group consisting of cells and proteins.
8. The non-adhesive biological material according to any one of claims 1 to 7, which is used for coating a medical device or coating a cell culture container.
A composition for forming a biological substance non-adhesive material, comprising a monomer represented by the formula (1) and a solvent.

In the formula (1), R 31 represents a hydrogen atom or a methyl group, a plurality of R 31 may be the same or different, and R 32 is a linear or branched alkylene group. And a plurality of R 32 may be the same or different from each other; R 33 represents a linear or branched alkylene group; and a plurality of R 33 may be the same as each other. M may be different, and m represents an integer of 3 to 8.
A compound represented by the formula (3).

In the formula (3), R 31 represents a hydrogen atom or a methyl group, a plurality of R 31 may be the same or different, and m represents an integer of 3 to 5.