JP2011084595A - Fluorine-containing ethylene oxide copolymer - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a fluorine-containing ethylene oxide copolymer which has an effect of inhibiting adhesion of a biotissue or a cell and an effect of inhibiting adhesion of a protein on a base material containing silica used particularly in a membrane, a filter, or the like, and has a biocompatibility, in medical and biomedical materials. <P>SOLUTION: The fluorine-containing ethylene oxide copolymer is an ethylene oxide copolymer having 0.1-50 mol% of a structural unit represented by formula (I) and 50-99.9 mol% of a structural unit represented by formula (II), in which a weight average molecular weight based on polyethylene oxide is 1,000 to 400,000. (In formula (I), R<SP>1</SP>represents a particular fluorine-containing group). <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a fluorine-containing ethylene oxide copolymer and use thereof.

  In recent years, medical and medical materials using various polymer materials have been studied, such as artificial kidney membranes, plasma separation membranes, virus removal membranes, artificial lung membranes, artificial blood vessels, adhesion prevention membranes, and wound coatings. Expected to be used for materials and artificial skin. In these fields, a synthetic material that is a foreign substance for a living body is used in contact with a living tissue or body fluid. Therefore, these medical and medical materials are required to have body fluid compatibility and body tissue compatibility so as not to cause interaction and interference with body tissues and body fluids.

  Body fluid compatibility and biological tissue compatibility required for medical and medical materials differ depending on the purpose and usage. For example, when used as a material in contact with blood, characteristics such as plasma protein adhesion inhibition, cell adhesion inhibition, blood coagulation system inhibition, platelet adhesion / activation inhibition and complement system activation inhibition are required. For this purpose, blood compatibility has been improved and various acrylate-based biocompatible materials have been reported (for example, Non-Patent Document 1 and Non-Patent Document 2).

  In rapid diagnosis using blood, the use of filters, non-woven fabrics or membranes with immobilized antibodies and proteins to capture the target component allows the capture of the target component simultaneously with the separation and removal of specific blood components. Or the function which can perform a coupling | bonding is calculated | required. In particular, there is a demand for suppression of non-specific adsorption, which is an obstacle to diagnosis, for substrates such as glass filters and nonwoven fabrics composed of silica materials, such as plasma protein adhesion suppression and cell adhesion suppression, which are the main causes. Function is required.

  As a functional material, an ethylene oxide copolymer, which is a biocompatible material other than acrylate, has been reported. In addition, various proposals have been made so far regarding copolymers having biofluidity and biocompatibility by polymerization of alkyl glycidyl ethers (for example, Patent Document 1).

JP 2005-281665 A

M. Tanaka, T. Motomura, M. Kawada, T. Anzai, Y. Kasori, T. Shiroya, K. Shimura, M. Onishi, A. Mochizuki, Biomaterials, 21, 1471-1481 (2000) K. Ishikawa, T. Ueda, N. Nakabayashi, Polymer Journal, 22, 355-360 (1990) TSE-LOK HO, Hard and Soft Acid and Bases Principle in Organic Chemistry, Academic Press, New York, 1977

  However, in Patent Document 1, for example, a material having a specific effect on a base material containing silica as a constituent material, such as a glass filter, has not been studied. In the medical and medical materials used for the parts that come into contact with body fluids or biological tissues, especially for base materials containing silica used for glass filters, membranes, etc., improved wettability to the base materials, suppression of tissue adhesion and tissue adhesion, Although suppression of plasma protein adhesion, suppression of cell adhesion, suppression of blood coagulation system, suppression of platelet adhesion / activation and suppression of complement system activation are particularly desired, no effective materials have been reported yet.

  The present invention has a biocompatibility in medical and medical materials, particularly with respect to a substrate containing silica, which is used for membranes, filters, etc., and has a biological tissue and cell adhesion inhibitory effect and a protein adhesion inhibitory effect. An object is to provide a fluorine-containing ethylene oxide copolymer and use thereof.

  The present invention provides biocompatibility in medical and medical materials, particularly for imparting biological tissue and cell adhesion suppression and protein adhesion suppression effects to substrates containing silica used in membranes, filters, and the like. A fluorine-containing ethylene oxide copolymer is provided.

  In the field of organosilicon chemistry, the silica cation is known as a hard acid, and the fluorine anion is known as a hard base. It is known to be stronger than “silica-oxygen bond” (Non-patent Document 3).

  Therefore, the present inventor believes that a stronger coating or addition / mixing is facilitated by allowing a polymer having a side chain containing fluorine atoms to act on a substrate containing silica as a constituent component. As a result, the present invention has been completed.

  Here, the silica in the present invention is a silicon compound having a siloxane bond portion or a silanol group on at least the surface. The main component of silica is silicon dioxide, but the “base material containing silica” described in this specification may contain alumina, sodium aluminate, etc. as a minor component, and sodium hydroxide as a stabilizer. Further, it may contain an inorganic base such as potassium hydroxide, lithium hydroxide or ammonia, or an organic base such as tetramethylammonium.

The present invention is as follows.
[1] An ethylene oxide copolymer having 0.1 to 50 mol% of a structural unit represented by the following formula (I) and 50 to 99.9 mol% of a structural unit represented by the following formula (II). And a fluorine-containing ethylene oxide copolymer having a weight average molecular weight in terms of polyethylene oxide of 1,000 to 400,000.
(In formula (I), R ¹ represents a monovalent group represented by the following formula (III).)
(In formula (III), m and n are integers that satisfy the conditions of 0 ≦ m ≦ 4 and 1 ≦ n ≦ 15, respectively, and R ² represents a fluorine atom or a hydrogen atom.)
[2] The fluorine-containing ethylene oxide copolymer of [1], wherein the ratio of the weight average molecular weight in terms of polyethylene oxide to the number average molecular weight in terms of polyethylene oxide is 1.1 to 2.7.
[3] Use of the fluorine-containing ethylene oxide copolymer of [1] or [2] for a portion that comes into contact with a biological tissue or body fluid.
[4] Use of the fluorine-containing ethylene oxide copolymer of [1] or [2] for a portion that contacts a peptide or protein.
[5] An additive to a resin material used in one or more applications selected from the group consisting of medical devices, biological protein purification, biological glycoprotein purification, and peptide purification, wherein [1] or [2] An additive comprising a fluorine-containing ethylene oxide copolymer.
[6] The additive according to [5], wherein the resin material is a resin material composed of one or more selected from the group consisting of polyester, polyamide, polyimide, polyether, polyolefin, polysulfone, silica, and polyfluorocarbon.
[7] A coating material for a resin material used for one or more applications selected from the group consisting of medical devices, biological protein purification, biological glycoprotein purification, and peptide purification, and the fluorine of [1] or [2] The coating agent which consists of a content ethylene oxide copolymer.
[8] The coating material according to [7], wherein the resin material is a resin material selected from the group consisting of polyester, polyamide, polyimide, polyether, polyolefin, polysulfone, silica, and polyfluorocarbon.
[9] A fiber treatment agent containing the fluorine-containing ethylene oxide copolymer of [1] or [2].
[10] A contact lens storage solution or cleaning solution containing the fluorine-containing ethylene oxide copolymer of [1] or [2].
[11] An additive or coating agent for a resin material used for diagnosis using body fluid or biological tissue, the additive or coating agent containing the fluorine-containing ethylene oxide copolymer of [1] or [2].
[12] A membrane containing silica, which is used for purifying or diagnosing one or more of the fluorine-containing ethylene oxide copolymers of [1] or [2] selected from the group consisting of body fluid, biological tissue, biologically derived protein and peptide A method for producing medical and medical members, comprising a step of coating one or more medical and medical members selected from the group consisting of a substrate, a filter and beads.
[13] A membrane containing silica, which is used for purification or diagnosis of one or more selected from the group consisting of body fluid, biological tissue, biological protein and peptide of the fluorine-containing ethylene oxide copolymer of [1] or [2] A method for producing medical and medical members, comprising the step of adding or mixing to a resin material constituting one or more medical and medical members selected from the group consisting of a substrate, a filter, and beads.

  According to the present invention, in medical and medical materials, it has a biological tissue and cell adhesion inhibitory effect and a protein adhesion inhibitory effect, especially on a substrate containing silica used for membranes, filters, etc., and has biocompatibility. The fluorine-containing ethylene oxide copolymer which has, its use, etc. can be provided.

It is a plot figure which shows the adsorption inhibitory effect of the human immunoglobulin G with respect to the 96-well EIA plate which coat | covered the copolymers (2)-(4) of Examples 2-4, respectively. It is a plot figure which shows the adsorption | suction suppression effect of human albumin with respect to the 96-well EIA plate which coat | covered the copolymers (2)-(4) of Examples 2-4, respectively. It is a plot figure which shows the adsorption | suction suppression effect of the human immunoglobulin G with respect to the glass filter which coat | covered the copolymers (2) of Example 2 and 4 and manufacture example 1, (4), and (6), respectively. It is a plot figure which shows the adhesion inhibitory effect of the HEL human lung cell with respect to the 96-well EIA plate which coat | covered the copolymers (2)-(4) of Examples 2-4, respectively.

  Hereinafter, a form for carrying out the present invention (hereinafter simply referred to as “the present embodiment”) will be described in detail with reference to the drawings as necessary. However, the present invention is not limited to the following embodiment. The present invention can be variously modified without departing from the gist thereof.

  The fluorine-containing ethylene oxide copolymer of the present embodiment (hereinafter also simply referred to as “copolymer”) has the structural units represented by the above formulas (I) and (II), preferably random copolymer It is a coalescence.

In formula (I), R ¹ represents a monovalent group represented by the above formula (III). In the formula (III), m and n are integers satisfying the conditions of 0 ≦ m ≦ 4 and 1 ≦ n ≦ 15, m is preferably 1, n is preferably 2 to 8, and 2 to 6 Is more preferable. R ² represents a fluorine atom or a hydrogen atom.

  In the copolymer of this embodiment, the content ratio of the structural unit represented by the above formula (I) and the structural unit represented by the above formula (II) is such that the structural unit represented by the above formula (I) is 0. 0.1 to 50 mol%, preferably 0.1 to 20 mol%, and the structural unit represented by the formula (II) is 50 to 99.9 mol%, preferably 80 to 99.9 mol%. The structural unit represented by the above formula (I) is 0.1 mol% or more, or the structural unit represented by the above formula (II) is 99.9 mol% or less. Thus, the affinity can be imparted more effectively. On the other hand, when the structural unit represented by the above formula (I) is 50 mol% or less, or the structural unit represented by the above formula (II) is 50 mol% or more, further suppression of adhesion of living tissue or cells. And the effect of protein adhesion suppression is acquired. The content ratio of these structural units is adjusted by the charging ratio of the monomer corresponding to each structural unit.

  The molecular weight of the fluorine-containing ethylene oxide copolymer of this embodiment is 1000 to 400,000 in terms of weight average molecular weight in terms of polyethylene oxide (PEO). When the weight average molecular weight is less than 1000, the copolymer becomes extremely soluble in water, but the excretion rate from the body becomes extremely high. On the other hand, when the weight average molecular weight exceeds 400,000, the copolymer is soluble in water, but the viscosity increases. Considering safety to living bodies from these viewpoints, the weight average molecular weight of the copolymer is preferably 1000 to 150,000, and more preferably 1000 to 100,000. Judging from suppression of blood retention and ease of renal excretion, the weight average molecular weight is more preferably 1000 to 70000. This is because when the weight average molecular weight of plasma albumin in the living body is lower than the weight average molecular weight (about 67,000), the excretion of the copolymer outside the body proceeds rapidly. Therefore, it is particularly preferable that the weight average molecular weight is 1000 to 70000 which is highly safe to living bodies while being controlled to be excreted outside the body.

  Here, the weight average molecular weight in terms of PEO means that when molecular weight is measured by GPC (gel permeation chromatography), polyethylene oxide (TSK standard Polyethylene Oxide, manufactured by TOSOH Co., Ltd.) is used as a standard sample, and its weight average. The molecular weight obtained in terms of molecular weight. Since PEO is also soluble in water-soluble solvents and organic solvents, it was used as a molecular weight standard substance when measuring the molecular weights of hydrophilic polymers and hydrophobic polymers.

  Moreover, since the weight average molecular weight is 1000-400000, since it melt | dissolves in either of a wide variety of organic solvents including water and ethanol, it is preferable. On the other hand, when the weight average molecular weight is less than 1000, the adsorption stability of the coating agent containing the same is lowered, so that it may be peeled off from the substrate surface to be coated. Further, when the weight average molecular weight exceeds 400,000, the viscosity of the fluorine-containing ethylene oxide copolymer solution becomes high, so that the dispersion or copolymer of the base particles when the base is added to the copolymer solution It may be difficult to prepare a particle surface having the above characteristics uniformly. From such a viewpoint, the weight average molecular weight is more preferably 1000 to 200000, and still more preferably 1000 to 100000.

  The fluorine-containing ethylene oxide copolymer of this embodiment is preferably a random copolymer in order to randomly disperse the hydrophilic portion and the hydrophobic portion in the same molecule. If the copolymer of the present embodiment is a block copolymer, a polyether that acts as a surfactant may be generated. When such a copolymer is taken into a living body, there is a possibility that a hemolytic action may be exhibited, which is less preferable for a living body than using a random copolymer.

  In the copolymer of this embodiment, the ratio of the weight average molecular weight (Mw) in terms of PEO to the number average molecular weight (Mn) in terms of PEO, that is, the molecular weight distribution represented by weight average molecular weight / number average molecular weight (Mw / Mn). Is preferably 1.1 to 2.7, since uniformity of quality is important in order to more fully exhibit the target tissue compatibility. In order to ensure a higher quality with respect to the copolymer, the molecular weight distribution is more preferably 1.1 to 2.5, still more preferably 1.1 to 2.2, and 1.1 to 1 .8, particularly preferably 1.0 to 1.5.

  The copolymer of the present embodiment is preferably produced using a monomer having a fluorine-containing group introduced as a monomer. Thereby, a fluorine-containing group can be directly introduced into the polymer in a one-step polymerization process. Furthermore, it is also preferable in that the amount of fluorine-containing group introduced can be controlled by adjusting the amount of monomer used with respect to ethylene oxide.

  As described above, the fluorine-containing ethylene oxide copolymer of the present embodiment, which is preferably obtained by random polymerization using a monomer having a fluorine-containing group, is excellent in various respects as compared with PEO which is the prior art. That is, in the copolymer of this embodiment, since a fluorine-containing group is introduced into the side chain, polyester, polyamide, polyimide, polyether, polyolefin, poly, in addition to silica particularly used for membranes and filters. It can act effectively on a substrate made of a resin material containing sulfone or polyfluorocarbon. The fluorine-containing ethylene oxide copolymer of the present embodiment has improved affinity for the substrate due to the effect of fluorine atoms in the side chain. Thereby, the copolymer of the present embodiment is improved in wettability, tissue adhesion and tissue adhesion inhibition, plasma protein adhesion inhibition, cell adhesion inhibition, blood coagulation system inhibition, platelet adhesion / activation inhibition and As a material for suppressing the activation of the complement system, it can be used as an effective material for coating on the surface of the substrate, or for addition to and mixing with the substrate.

  Furthermore, the fluorine-containing ethylene oxide copolymer of the present embodiment can be applied to electronic materials such as DNA substrates and silicon substrates such as glass substrates for small molecule microarrays and electronic substrates due to the effect of fluorine atoms in the side chain. In addition, antifouling effects such as protein adsorption inhibition and biological tissue adhesion inhibition can be exhibited. Here, examples of the “board” include an electronic circuit board, a printed wiring board, and a module board.

  About the monomer composition of the copolymer of the present embodiment, the type of raw material used in the reaction, the mass of the raw material, the type of polymerization initiator, the addition amount of the polymerization initiator, the presence or absence of the reaction solvent, the selection of the reaction solvent, the reaction temperature, the reaction Copolymers having various composition ratios are produced by adjusting time, raw material concentration, polymerization initiator concentration, reaction atmosphere, reaction pressure, stirring state, stirring speed, and the like. For example, in a reaction solvent or in the absence of a solvent, one or more glycidyl ethers together with ethylene oxide are used alone or in combination with a metal polymerization initiator or other polymerization initiator, and argon, nitrogen, etc. The copolymer of this embodiment is obtained by ring-opening polymerization under an inert gas atmosphere, ice-cooling, room temperature, or heating as necessary under normal pressure or pressure.

  Examples of the reaction solvent include toluene, hexane, heptane, 2-methoxyethyl ether, ethylene glycol dimethyl ether, and ethylene glycol diethyl ether. Examples of glycidyl ethers include 3- (2,2,3,3-tetrafluoropropoxy) -1,2-propene oxide, 3- (1H, 1H, 5H-octafluoropentyloxy) -1,2- Propene oxide, 3- (1H, 1H, 7H-dodecafluoroheptyloxy) -1,2-propene oxide, 3- (1H, 1H, 9H-hexadecafluorononyloxy) -1,2-propene oxide, 3- (1H, 1H-pentafluoropropoxy) -1,2-propene oxide, 3- (1H, 1H-nanofluoropentyloxy) -1,2-propene oxide, 3- (1H, 1H-tridecafluoroheptyloxy) -1,2-propene oxide, 3- (1H, 1H-heptadecafluorononyloxy) -1,2-propene oxide It is below. Examples of the metal polymerization initiator include Lewis acids such as boron trifluoride ether complex, trialkylaluminum, triethylaluminum, triphenylaluminum, and tributylaluminum. Other polymerization initiators include, for example, potassium hydroxide, sodium hydroxide, cesium hydroxide, sodium methoxide, sodium ethoxide, sodium propoxide, sodium t-butoxide, potassium propoxide, potassium t-butoxide, potassium -T-2-methyl-2-butoxide.

  As described above, it is important that the weight average molecular weight of the copolymer in the present embodiment is a molecular weight that can be easily excreted from the living body to the outside of the body from the viewpoint of safety. For example, in order to set the weight average molecular weight in terms of PEO in GPC to 1000 to 400,000, a known method, for example, purification of starting materials, fractional removal of a high molecular weight fraction or a low molecular weight fraction after the reaction is performed as necessary. It may be adopted. For example, specific methods for molecular weight fractionation include size exclusion chromatography (SEC), chromatography such as GPC, ultrafiltration including UF module, ultracentrifugation, organic solvents such as ether and ethanol, etc. The precipitation fraction used is mentioned.

  As described above, for example, in the field of medical and medical materials in which the copolymer of the present embodiment is used for a part in contact with a body fluid or biological tissue, in addition to silica used for membranes and filters, polyester, polyamide, polyimide When used for a base material made of resin materials including polyether, polyolefin, polysulfone or polyfluorocarbon, select the optimal molecular weight and introduce the optimal amount of side chain fluorine-containing groups according to the purpose of use. It becomes possible to do.

  The present embodiment relates to a silica-affinity polymer comprising a fluorine-containing ethylene oxide copolymer that can be used for a part that comes into contact with a body fluid or a biological tissue, and a part that comes into contact with a peptide or a protein. It is. The fluorine-containing ethylene oxide copolymer of the present embodiment has not only a biological tissue adhesion and adhesion suppression effect, a protein adhesion suppression effect and a cell adhesion suppression effect, but also silica, polyester, polyamide, polyimide, polyether, polyolefin, polysulfone. Or it has the improvement effect of the wettability with respect to the base material which consists of a resin material containing polyfluorocarbon, the adhesion effect of platelets, and the suppression effect of activation.

  The fluorine-containing ethylene oxide copolymer of this embodiment can be used for a part that comes into contact with a body fluid or a biological tissue, and a part that comes into contact with a peptide or protein, and can be used for a medical device. In addition, the fluorine-containing ethylene oxide copolymer of the present embodiment has an effect of suppressing interaction and interference with body fluids, biological tissues, peptides, and proteins, and suppresses adhesion and adhesion of biological tissues, plasma protein It has an adhesion inhibitory effect, a cell adhesion inhibitory effect, a platelet adhesion and activation inhibitory effect, a complement system activation inhibitory effect, a peptide adhesion inhibitory effect, and a protein adhesion inhibitory effect. Furthermore, the copolymer of the present embodiment improves the recovery rate of the target peptide and protein by mixing or coating the material constituting the equipment for the purpose of separation and purification of peptides and proteins. In addition, the intended effect sustainability and effect sustainability of the equipment can be achieved. Therefore, the fluorine-containing ethylene oxide copolymer of the present embodiment is an additive to a resin material used for one or more applications selected from the group consisting of medical devices, biological protein purification, biological glycoprotein purification and peptide purification. Alternatively, it is useful as a coating agent for the resin material.

  The copolymer of the present embodiment is used for, for example, medical use as an artificial kidney membrane, plasma separation membrane, artificial lung membrane, artificial blood vessel, adhesion prevention membrane, wound dressing material, artificial skin, virus removal membrane, leukocyte removal membrane. It can be used as a constituent material or a coating agent of biomaterials such as medical materials, medical materials, and medical devices. The copolymer of this embodiment further covers wounds due to bed sores, burns, ulcers, trauma, or wounds caused by tissue destruction that covers the dermis, subcutaneous tissue, muscles, muscles, joints, and bones in the form of a film. Can be used to Furthermore, the copolymer of this embodiment can be used also as a contact lens storage solution or cleaning solution by utilizing the hydrophilicity imparting effect (moisturizing effect) and the protein adsorption inhibiting effect and exhibiting an antifouling effect. In addition, the copolymer of the present embodiment can also exert an antifouling effect on electronic materials such as electronic substrates by utilizing the protein adsorption inhibiting effect on DNA and glass substrates for small molecule microarrays or silica substrates. . Furthermore, the copolymer of this embodiment can be used also for fiber treatments, such as cosmetics or a nonwoven fabric, utilizing the hydrophilicity provision and moisture retention provision effect.

  On the other hand, according to the present embodiment, with respect to a substrate made of silica used for a membrane, a filter, or the like, the wettability is specifically improved by the effect of improving the silica affinity by the side chain containing fluorine atoms, and the biological tissue It is possible to impart an adhesion and adhesion inhibiting effect, a protein adhesion inhibiting effect, a cell adhesion inhibiting effect, a platelet adhesion and activation inhibiting effect.

  The copolymer of the present embodiment can be used for separation, purification, concentration, filtration, desalting and concentration of biological peptides or proteins including humans, mammals, reptiles, microorganisms, insects and the like. Alternatively, it can be used for separating, purifying, concentrating, filtering, desalting and the like of pharmaceuticals, pharmaceutical active ingredients, and pharmaceutical raw materials comprising the peptide or protein. Moreover, the copolymer of this embodiment can be used as an additive with respect to the base material which comprises the equipment for using for these uses, or a coating agent with respect to the equipment. Furthermore, the copolymer of the present embodiment utilizes a biological non-recognition action such as its moisturizing action and protein adsorption inhibiting action, as a cosmetic, for example, a hair cosmetic for the purpose of suppressing the charging of hair, It can be used as an irritation inhibitor for fibers that can reduce irritation of a fiber material to which an irritating substance is attached.

  According to the present embodiment, the fluorine-containing ethylene oxide copolymer is a film, substrate, or filter containing silica used for purification or diagnosis of one or more selected from the group consisting of body fluid, biological tissue, biologically derived protein, and peptide. In addition, by coating one or more known medical and medical members selected from the group consisting of beads, medical and medical members that exhibit the above effects can be manufactured. Further, according to the present embodiment, the fluorine-containing ethylene oxide copolymer is a film or substrate containing silica that is used for purification or diagnosis of one or more selected from the group consisting of body fluid, biological tissue, biological protein and peptide Producing a medical and medical member having the above-mentioned effects by adding or mixing to one or more known medical and medical members that may be selected from the group consisting of a filter and a bead Can do.

EXAMPLES Hereinafter, although an Example demonstrates this invention further more concretely, the scope of the present invention is not limited to the following Example. Moreover, the conditions of GPC in a present Example are as follows.
Column: G40000PWXL (manufactured by Tosoh Corporation)
Mobile phase: 20% acetonitrile / 50 mM lithium chloride Flow rate: 0.8 mL / min Column temperature: 40 ° C.
GPC device: LC-10A system (manufactured by Shimadzu Corporation)
Detector: RID-10A (RI: differential refractometer, manufactured by Shimadzu Corporation)
The calibration curve for calculating the molecular weight and molecular weight distribution of the compound was standard polyethylene oxide (manufactured by Tosoh Corporation, weight average molecular weight: 2.4 × 10 ⁴ , 5.00 × 10 ⁴ , 10.7 × 10 ⁴ , 14. 0 × 10 ⁴ ).

Example 1
Under an argon atmosphere, in a pressure-resistant reaction vessel, 2- (2,2,3,3-tetrafluoropropoxy) -1,2-propoxide 22.4 g, ethylene oxide 80 mL, potassium t-butoxide 1 M tetrahydrofuran solution 1 and triisobutylaluminum 1 M hexane solution 10 mL and 300 mL of hexane as a solvent were added, and the reaction was performed at 25 ° C. for 24 hours. After completion of the reaction, the reaction product was recovered, and the solvent was removed under reduced pressure to obtain 71 g of the desired random copolymer (1) as a white solid. The result of having measured the weight average molecular weight of PEO conversion of the obtained fluorine-containing ethylene oxide copolymer by GPC was 51000. Introduction rate of fluorine-containing groups (structural units represented by the above formula (I), the same applies hereinafter) by ¹ H-NMR analysis in a deuterated methanol (CD ₃ OD) solvent using tetramethylsilane as a standard. (Content ratio; the same applies hereinafter) was 4.5 mol%.

(Example 2)
Under an argon atmosphere, in a pressure-resistant reaction vessel, 3- (1H, 1H, 5H-Octafluoropentyloxy) -1,2-propoxide 13.6 g, ethylene oxide 41 mL, potassium t-butoxide 1M tetrahydrofuran solution 0.5 mL and triisobutylaluminum 1M hexane solution 4.5 mL and 200 mL of hexane as a solvent were added and reacted at 25 ° C. for 20 hours. After completion of the reaction, the reaction product was recovered, and the solvent was removed under reduced pressure to obtain 41 g of the desired random copolymer (2) as a white solid. The result of measuring the weight average molecular weight in terms of PEO of the obtained fluorine-containing ethylene oxide copolymer by GPC was 46000. The introduction rate of fluorine-containing groups was 3.2 mol% by ¹ H-NMR analysis in a deuterated methanol (CD ₃ OD) solvent using tetramethylsilane as a standard.

(Example 3)
Under an argon atmosphere, in a pressure resistant reactor, 3- (1H, 1H, 7H-Dodecafluoroxy) -1,2-propenoxide 13.9 g, 41.3 mL of ethylene oxide, 4.5 mL of 1 molar hexane solution of triisobutylaluminum and potassium t -Butoxide 56mg and hexane 400mL were added as a solvent, and reaction was performed at 25 degreeC for 20 hours. After completion of the reaction, the solvent was removed to obtain 36 g of the desired random copolymer (3) as a white solid. The result of measuring the weight average molecular weight in terms of PEO of the obtained fluorine-containing ethylene oxide copolymer by GPC was 43,000. The introduction rate of the fluorine-containing group was 2.9 mol% by ¹ H-NMR analysis in a deuterated methanol (CD ₃ OD) solvent using tetramethylsilane as a standard.

Example 4
Under an argon atmosphere, in a pressure-resistant reaction vessel, 9.3 g of 3- (2,2,3,3-tetrafluoropropoxy) -1,2-propenoxide, 70 mL of ethylene oxide, 1.1 mL of potassium 2-methyl-2-butoxide 1M tetrahydrofuran solution and Triisobutylaluminum 1M hexane solution 11.1mL and hexane 300mL as a solvent were added, and reaction was performed at 25 degreeC for 6 hours. After completion of the reaction, the reaction product was collected and the solvent was removed under reduced pressure to obtain 49 g of the desired random copolymer (4) as a white solid. The weight average molecular weight in terms of PEO of the obtained fluorine-containing ethylene oxide copolymer was measured by GPC to be 49000. The introduction rate of fluorine-containing groups was 2.9 mol% by ¹ H-NMR analysis in a deuterated methanol (CD ₃ OD) solvent using tetramethylsilane as a standard.

(Example 5)
Under an argon atmosphere, in a pressure-resistant reaction vessel, 9.3 g of 3- (2,2,3,3-tetrafluoropropoxy) -1,2-propenoxide, 70 mL of ethylene oxide, 1 mL of potassium t-butoxide 1M tetrahydrofuran solution and triisobutylaluminum 1M hexane solution 10 mL and 300 mL of hexane as a solvent were added, and the reaction was performed at 30 ° C. for 4.5 hours. After completion of the reaction, the reaction product was collected and the solvent was removed under reduced pressure to obtain 50 g of the desired random copolymer (5) as a white solid. The result of having measured the weight average molecular weight of PEO conversion of the obtained fluorine-containing ethylene oxide copolymer by GPC was 35000. The introduction rate of fluorine-containing groups was 2.2 mol% by ¹ H-NMR analysis in a deuterated methanol (CD ₃ OD) solvent using tetramethylsilane as a standard.

(Production Example 1)
As a comparative control, a random copolymerized ethylene oxide copolymer containing no fluorine atom was produced with reference to JP-A-2005-281665. Under an argon atmosphere, n-butyl glycidyl ether 14.2 mL, ethylene oxide 41.3 mL, triisobutylaluminum 1 M hexane solution 4.5 mL and potassium t-butoxide 1 M THF solution 0.5 mL in a pressure-resistant reaction vessel, and hexane 200 mL as a solvent And reacted at 25 ° C. for 20 hours. After completion of the reaction, the solvent was removed to obtain 46 g of the desired copolymer (6) as a white solid. The result of measuring the weight average molecular weight in terms of PEO of the obtained copolymer by GPC was 67,000. According to ¹ H-NMR analysis, multiple line peaks derived from n-butyl groups were observed at δ0.94, δ1.39, and δ1.54 by measurement in a CD ₃ OD solvent using tetramethylsilane as a standard. It was. Furthermore, a peak mainly derived from polyethylene glycol was observed at δ 3.30 to 3.90. Moreover, the introduction rate of n-butyl group by ¹ H-NMR analysis was 2.8 mol%.

(Example 6)
Copolymers (2), (3) and (4) in the above examples were respectively added to a 70% aqueous ethanol solution at concentrations of 10, 1, 0.1, 0.01, 0.001 and 0 mg / mL. A sample solution was obtained by dissolution. By adding the above sample solution at a rate of 0.2 mL / well to a Coaster 96-well EIA plate (product number 3590) manufactured by Corning, Inc. (Corning, NY, USA), and allowing to stand at 4 ° C. overnight. Covered. Similarly, a 70% aqueous ethanol solution was added to the control well. Thereafter, the sample solution was removed by suction, and the plate was dried to produce a plate coated with the copolymer. This plate was subjected to plasma protein adsorption evaluation by the following method.

  5 μg / mL solution of purified human immunoglobulin G (ICN / Cappel, USA) manufactured by Capel Co., Ltd. was added to the EIA plate at a rate of 0.1 mL / well, and the well surface was 37 ° C. For 2 hours. Subsequently, the amount of human immunoglobulin G adsorbed on the well surface was determined by using the horseradish peroxidase (HRP) -conjugated goat IgG fraction to human IgG (Whole molecule ICN / Cappel, USA)), and was evaluated by enzyme-linked immunosorbant assay (ELISA).

  The evaluation results are shown in FIG. FIG. 1 shows copolymer (2) of Example 2 (here, denoted as C (2)), copolymer (3) of Example 3 (here, denoted as C (3)), and Example 4 It is a plot figure which shows the adsorption inhibitory effect of the human immunoglobulin G with respect to the 96-well EIA plate each coat | covered with the copolymer (4) (here, it describes with C (4)). A calibration curve was prepared by adding an immunoglobulin standard of known concentration to the same plate and measuring. Based on this calibration curve, the amount of immunoglobulin G adsorbed on the plate was quantified.

  As shown in FIG. 1, the 96-well EIA plate coated with the copolymer (2) of Example 2, the copolymer (3) of Example 3 and the copolymer (4) of Example 4 was coated. The amount of immunoglobulin G adsorbed to the uncoated well was approximately 5 μg / mL, and the test sample showed a tendency to suppress human immunoglobulin G adsorption depending on the amount of coating.

(Example 7)
Copolymers (2), (3) and (4) in the above examples were respectively added to a 70% aqueous ethanol solution at concentrations of 10, 1, 0.1, 0.01, 0.001 and 0 mg / mL. A sample solution was obtained by dissolution. By adding the above sample solution at a rate of 0.2 mL / well to a Coaster 96-well EIA plate (product number 3590) manufactured by Corning, Inc. (Corning, NY, USA), and allowing to stand at 4 ° C. overnight. Covered. Similarly, a 70% aqueous ethanol solution was added to the control well. Thereafter, the sample solution was removed by suction, and the plate was dried to prepare a plate coated with the above compound. This plate was subjected to plasma protein adsorption evaluation by the following method.

  A 2 μg / mL solution of purified human albumin (Purified Human Albumin; ICN / Cappel, USA) manufactured by Kapell is added to a 96-well EIA plate at a rate of 0.1 mL / well, and the well surface is contacted at 37 ° C. for 2 hours. I let you. Then, the amount of human albumin adsorbed on the well surface was determined using ELISA using Horseradish peroxidase (HRP) -conjugated goat IgG fraction to human albumin (ICN / Cappel, USA). It was evaluated by.

  The evaluation results are shown in FIG. FIG. 2 shows copolymer (2) of Example 2 (denoted here as C (2)), copolymer (3) of Example 3 (denoted as C (3) here), and Example 4 It is a plot figure which shows the adsorption | suction suppression effect of the human albumin with respect to the 96-well EIA plate which each coat | covered the copolymer (4) (here, it describes with C (4)). A standard curve was prepared by adding a standard human albumin standard of known concentration to the same plate and measuring. As a result, 96-hole EIA plates respectively coated with the copolymer (2) of Example 2, the copolymer (3) of Example 3 and the copolymer (4) of Example 4 are shown in FIG. As can be seen, the amount of immunoglobulin G adsorbed to the uncoated well was approximately 2 μg / mL, and the test sample showed a tendency to suppress the adsorption of human immunoglobulin G according to the amount of coating.

(Example 8)
Plasma protein adsorption evaluation was performed by the following method using a glass filter having a diameter of 8 mm (filter paper manufactured by Kiriyama Seisakusho, GFP 8 mmφ) instead of the EIA plate in Example 6. That is, the copolymers (2), (4) and (6) in the above examples and polyethylene glycol (molecular weight 20000, PEG 20000, manufactured by Wako Pure Chemical Industries) as a control, Sample solutions were obtained by dissolving at concentrations of 0.1, 0.01, 0.001 and 0 mg / mL. Place a glass filter with a diameter of 8 mm, washed with methanol and dried in a 24-well plate for cell culture, add 50 μL of the sample solution to the glass filter, and dry under reduced pressure. Produced. This glass filter was evaluated for plasma protein adsorption by the following method.

  A 5 μg / mL solution of purified human immunoglobulin G (Purified Human IgG; ICN / Cappel, USA) manufactured by Capel Co., Ltd. was added to a 0.4-well plate in a 24-well plate containing a glass filter coated with the copolymer. Wells were added in proportions and the glass filters were contacted at 37 ° C. for 2 hours. Thereafter, the amount of human immunoglobulin G adsorbed on the glass filter was measured by using horseradish peroxidase (HRP) -conjugated goat IgG fraction to human IgG (wholemolecule; ICN / Cappel, USA)).

  The evaluation results are shown in FIG. FIG. 3 shows the copolymer (2) of Example 2 (indicated here as C (2)), the copolymer (4) of Example 4 (indicated here as C (4)), and Production Example 1 It is a plot figure which shows the adsorption | suction suppression effect of the human immunoglobulin G with respect to the glass filter which each coat | covered the copolymer (6) (here, it describes with C (6)). A calibration curve was prepared by adding a known concentration of immunoglobulin G standard to a 24-well plate and measuring. Based on this calibration curve, the amount of human immunoglobulin adsorbed on a glass filter placed in a 24-well plate was quantified.

  As a result, the glass filter coated with the copolymer (2) of Example 2, the copolymer (4) of Example 4 and the copolymer (6) of Production Example 1 was subjected to the coating amount. Inhibition of human immunoglobulin G adsorption was observed. On the other hand, a glass filter coated with PEG 20000 as a control had weak inhibition of human immunoglobulin G adsorption. Further, the glass filter coated with the copolymers (2) and (4) tended to have a higher human immunoglobulin G adsorption inhibitory effect than the glass filter coated with the copolymer (6).

Example 9
<Evaluation of human cell adhesion: Experiments for examining cell adhesion inhibitory effect and cytotoxicity>

  Copolymers (2), (3) and (4) in the above examples were added in 70% ethanol aqueous solution at concentrations of 5, 2.5, 1.0, 0.5, 0.1 and 0 mg / mL, respectively. To obtain a sample solution. This sample solution was added to each 96-well plate for cell culture at a rate of 0.2 mL / well and allowed to stand overnight at 4 ° C. to cover the inside of the plate well. Control wells were coated with 70% aqueous ethanol. Thereafter, the sample solution was removed by suction, and the plate was dried to produce a plate coated with the copolymer. Two plates each coated with the same copolymer were prepared.

  About these two plates, cell adhesion evaluation was performed with the following method.

HEL human lung cell solution (3 × 10 ⁵ cells / mL) was added to each 96-well plate coated with the test sample solution, and 0.1 mL / well was added thereto, followed by incubation for 2 hours. After completion of the culture, the number of viable cells attached to the plate was evaluated by CellTiter 96® AQueous Assay system (manufactured by Promega Corp. (Madison, Wis., USA)). Further, using another plate to evaluate the cytotoxicity of the copolymers (2), (3) and (4), in addition to the attached cells, the cells that did not adhere and floated, that is, Total cells added to each well were evaluated using the CellTiter 96 (R) AQueous Assay system under the same conditions as described above.

  The evaluation results are shown in FIG. FIG. 4 is a plot diagram showing the adhesion inhibitory effect of HEL human lung cells on 96-well EIA plates coated with the copolymers (2) to (4) of Examples 2 to 4, respectively. As shown in FIG. 4, each of the 96-well plates coated with the copolymer (2) of Example 2, the copolymer (3) of Example 3, and the copolymer (4) of Example 4 was used. When the cell adhesion amount to the control coated (polymer non-coated) well was taken as 100%, HEL human lung cell adhesion suppression tendency corresponding to the coating amount was observed for any copolymer (C in the figure). (Indicated by (2), C (3), C (4)).

  Further, regarding cytotoxicity, in the 96-well plate coated with the copolymer (2) of Example 2, the copolymer (3) of Example 3 and the copolymer (4) of Example 4, respectively, FIG. As shown, any copolymer had a viable cell count of 80% or more with respect to the control coating (polymer uncoated), and showed no cytotoxicity regardless of the coating amount (C ( 2) Indicated by *, C (3) *, C (4) *).

  According to the present invention, fluorine-containing ethylene that can effectively act on a substrate containing silica, particularly used in membranes and filters, in the field of medical and medical materials used in parts that come into contact with body fluids or biological tissues. An oxide copolymer can be provided. Due to the effect of fluorine atoms in the side chain in the fluorine-containing ethylene oxide copolymer of the present invention, the affinity for the substrate is improved, thereby improving wettability, suppressing tissue adhesion and adhesion, and suppressing plasma protein adhesion. , As a material for inhibiting cell adhesion, inhibiting blood coagulation system, inhibiting platelet adhesion / activation and inhibiting the activation of complement system, in particular, coating on the surface of the substrate, or addition to the substrate, The copolymer can be used as an effective material for mixing.

Claims (13)

An ethylene oxide copolymer having 0.1 to 50 mol% of a structural unit represented by the following formula (I) and 50 to 99.9 mol% of a structural unit represented by the following formula (II), A fluorine-containing ethylene oxide copolymer having a weight average molecular weight in terms of oxide of 1,000 to 400,000.
(In formula (I), R ¹ represents a monovalent group represented by the following formula (III).)
(In formula (III), m and n are integers that satisfy the conditions of 0 ≦ m ≦ 4 and 1 ≦ n ≦ 15, respectively, and R ² represents a fluorine atom or a hydrogen atom.)   The fluorine-containing ethylene oxide copolymer according to claim 1, wherein the ratio of the weight average molecular weight in terms of polyethylene oxide to the number average molecular weight in terms of polyethylene oxide is 1.1 to 2.7.   Use of the fluorine-containing ethylene oxide copolymer according to claim 1 or 2 for a portion that comes into contact with a biological tissue or body fluid.   Use of the fluorine-containing ethylene oxide copolymer according to claim 1 or 2 for a portion in contact with a peptide or protein.   The fluorine-containing ethylene according to claim 1 or 2, which is an additive to a resin material used for one or more uses selected from the group consisting of medical devices, biological protein purification, biological glycoprotein purification and peptide purification. An additive comprising an oxide copolymer.   The additive according to claim 5, wherein the resin material is a resin material composed of one or more selected from the group consisting of polyester, polyamide, polyimide, polyether, polyolefin, polysulfone, silica, and polyfluorocarbon.   A fluorine-containing ethylene oxide according to claim 1 or 2, which is a coating material for a resin material used for one or more applications selected from the group consisting of medical devices, biological protein purification, biological glycoprotein purification, and peptide purification. A coating comprising a copolymer.   The coating agent according to claim 7, wherein the resin material is a resin material composed of one or more selected from the group consisting of polyester, polyamide, polyimide, polyether, polyolefin, polysulfone, silica, and polyfluorocarbon.   The fiber processing agent containing the fluorine-containing ethylene oxide copolymer of Claim 1 or 2.   A contact lens preservation solution or cleaning solution comprising the fluorine-containing ethylene oxide copolymer according to claim 1.   An additive or coating agent for a resin material used for diagnosis using a body fluid or a biological tissue, comprising the fluorine-containing ethylene oxide copolymer according to claim 1 or 2.   A film, a substrate containing silica, used for purifying or diagnosing at least one selected from the group consisting of body fluid, biological tissue, biological protein and peptide, the fluorine-containing ethylene oxide copolymer according to claim 1 or 2; A method for producing medical and medical members, comprising a step of coating one or more medical and medical members selected from the group consisting of a filter and beads.   A film, a substrate containing silica, used for purifying or diagnosing at least one selected from the group consisting of body fluid, biological tissue, biological protein and peptide, the fluorine-containing ethylene oxide copolymer according to claim 1 or 2; A method for producing medical and medical members, comprising a step of adding or mixing to a resin material constituting one or more medical and medical members selected from the group consisting of filters and beads.