WO2013176084A1 - Silane coupling agent containing silylalkyl phosphoramidate compound - Google Patents

Silane coupling agent containing silylalkyl phosphoramidate compound Download PDF

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WO2013176084A1
WO2013176084A1 PCT/JP2013/063943 JP2013063943W WO2013176084A1 WO 2013176084 A1 WO2013176084 A1 WO 2013176084A1 JP 2013063943 W JP2013063943 W JP 2013063943W WO 2013176084 A1 WO2013176084 A1 WO 2013176084A1
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silane coupling
coupling agent
substrate
group
silylalkyl
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PCT/JP2013/063943
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French (fr)
Japanese (ja)
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義和 大塚
武久 岩間
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日産化学工業株式会社
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic System
    • C07F9/02—Phosphorus compounds
    • C07F9/06—Phosphorus compounds without P—C bonds
    • C07F9/22—Amides of acids of phosphorus
    • C07F9/24—Esteramides
    • C07F9/2454—Esteramides the amide moiety containing a substituent or a structure which is considered as characteristic
    • C07F9/2458—Esteramides the amide moiety containing a substituent or a structure which is considered as characteristic of aliphatic amines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/54—Biologically active materials, e.g. therapeutic substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L29/00—Materials for catheters, medical tubing, cannulae, or endoscopes or for coating catheters
    • A61L29/14—Materials characterised by their function or physical properties, e.g. lubricating compositions
    • A61L29/16—Biologically active materials, e.g. therapeutic substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L31/16—Biologically active materials, e.g. therapeutic substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L33/00—Antithrombogenic treatment of surgical articles, e.g. sutures, catheters, prostheses, or of articles for the manipulation or conditioning of blood; Materials for such treatment
    • A61L33/0005—Use of materials characterised by their function or physical properties
    • A61L33/0011—Anticoagulant, e.g. heparin, platelet aggregation inhibitor, fibrinolytic agent, other than enzymes, attached to the substrate
    • A61L33/0041—Anticoagulant, e.g. heparin, platelet aggregation inhibitor, fibrinolytic agent, other than enzymes, attached to the substrate characterised by the choice of an antithrombatic agent other than heparin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L33/00—Antithrombogenic treatment of surgical articles, e.g. sutures, catheters, prostheses, or of articles for the manipulation or conditioning of blood; Materials for such treatment
    • A61L33/04—Use of organic materials, e.g. acetylsalicylic acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic System
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6564—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms
    • C07F9/6571—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms having phosphorus and oxygen atoms as the only ring hetero atoms
    • C07F9/657163—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms having phosphorus and oxygen atoms as the only ring hetero atoms the ring phosphorus atom being bound to at least one carbon atom
    • C07F9/657181—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms having phosphorus and oxygen atoms as the only ring hetero atoms the ring phosphorus atom being bound to at least one carbon atom the ring phosphorus atom and, at least, one ring oxygen atom being part of a (thio)phosphonic acid derivative
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
    • A61L2300/216—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials with other specific functional groups, e.g. aldehydes, ketones, phenols, quaternary phosphonium groups
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/424—Anti-adhesion agents

Definitions

  • the present invention includes a silylalkyl phosphoramide compound capable of suppressing the adhesion of biological materials such as proteins, cells, and platelets by performing a surface treatment on an inorganic material such as silicon or glass, or a resin such as polyethylene.
  • the present invention relates to a silane coupling agent.
  • an anticoagulant such as heparin or a drug such as an immunosuppressant must be used in combination.
  • MPC polymer poly (2-methacryloyloxyethyl phosphorylcholine) (hereinafter referred to as MPC polymer) having phosphorylcholine, which is the same amphoteric phospholipid as the biological membrane, in the side chain of the polymer chain is used Medical materials that have been proposed have been proposed (see, for example, Patent Document 1 and Patent Document 2).
  • CMB polymer N-methacryloyloxyethyl-N, N-dimethylammonium- ⁇ -N-methylcarboxybetaine
  • An object of the present invention is a compound that is a non-polymer that is easy to produce, and the compound is directly applied to the surface of a substrate by applying the solution in which the compound is dissolved to the substrate or immersing the substrate in the solution. It is another object of the present invention to provide a compound capable of firmly covering the surface of the body and exhibiting the same biological substance adhesion inhibitory effect as the conventional MPC polymer and CMB polymer.
  • silylalkyl phosphoramidade compounds are useful as silane coupling agents, and silane coupling agents containing such compounds are proteins, cells, It has been found that it has an effect of suppressing adhesion of biological substances such as platelets. That is, the present invention 1. Following formula (1):
  • a silane coupling agent composition comprising the silylalkyl phosphoramidade compound according to 1 above, 3.
  • a silane coupling agent comprising a step of applying and baking the silane coupling agent composition described in 2 or 3 above on a substrate, a step of washing the substrate with a polar solvent, and a step of drying the substrate.
  • Immobilization method 5. Immobilization of a silane coupling agent comprising a step of immersing a substrate in the silane coupling agent composition described in 2 or 3 above, a step of washing the substrate with a polar solvent, and a step of drying the substrate.
  • Conversion method 6).
  • silane coupling agent surface treatment agent
  • a base material a resin such as polyethylene
  • the surface is coated with a silylalkyl phosphoramidade compound having both stable positive and negative charges in one molecule. Therefore, adhesion of biological substances such as proteins, cells, and platelets to the inorganic substance or resin surface can be suppressed without using a polymer such as MPC polymer or CMB polymer.
  • FIG. 1 is a diagram showing the cell adhesion inhibitory action of 2- (trimethylammonio) ethyl (3- (ethoxydimethylsilyl) propyl) phosphoramidate on a glass substrate.
  • Cell free represents a glass substrate treated with 2- (trimethylammonio) ethyl (3- (ethoxydimethylsilyl) propyl) phosphoramidate (30 mg / ml), and the control was controlled with the silane coupling agent of the present invention. Represents the result when no processing is performed.
  • FIG. 2 is a diagram showing the results of measurement of living cells attached to a glass substrate of a silylalkyl phosphoramidade compound. Control represents the result when the treatment with the silane coupling agent of the present invention is not performed.
  • silylalkyl phosphoramidade compound of the present invention represented by the formula (1) is 0.8 to 1.2 equivalents, preferably 1 equivalent of the formula (II) to the aminoalkylsilane represented by the formula (I). ) Is reacted at 20 to 80 ° C. in the presence of a base to obtain a compound represented by the formula (III). It can be obtained by reacting 8 to 1.2 equivalents, preferably 1 equivalent of a tertiary amine represented by the formula (IV) at 0 to 50 ° C.
  • an organic base such as triethylamine is preferable.
  • R 1 , R 2 and R 3 are methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl.
  • Group, isobutyl group, s-butyl group, t-butyl group, cyclobutyl group, n-pentyl group, and cyclopentyl group, and methyl group and ethyl group are particularly preferable.
  • R 4 is methyl group, ethyl group, n-propyl group, isopropyl group, cyclopropyl group, n-butyl group, isobutyl group, s -Butyl group, t-butyl group, cyclobutyl group, n-pentyl group, and cyclopentyl group can be mentioned, and methyl group and ethyl group are particularly preferable.
  • R 5 represents methyl group, ethyl group, n-propyl group, isopropyl group, cyclopropyl group, n-butyl group, isobutyl group, s -Butyl group, t-butyl group, cyclobutyl group, n-pentyl group, and cyclopentyl group can be mentioned, and methyl group and ethyl group are particularly preferable.
  • n may be an integer of 1 to 5, and 2 or 3 is particularly preferable.
  • p may be an integer of 1 to 5, but 2 or 3 is particularly preferable.
  • q is an integer of 1 to 3.
  • the silylalkyl phosphoramariae compound of the present invention is used for surface treatment of an inorganic substance as a base material
  • the inorganic substance is not particularly limited.
  • silicon, copper, iron, aluminum, zinc or an alloy thereof, glass, silica, aluminum oxide, aluminum hydroxide, and magnesium oxide can be given.
  • the resin is not particularly limited.
  • examples include poly (meth) acrylamide, poly (meth) acrylamide derivatives, polysulfone, polycarbonate, cellulose, and cellulose derivatives.
  • the silylalkyl phosphoramidade compound of the present invention can be used for surface treatment of pharmaceuticals, quasi drugs, medical instruments and the like.
  • medical devices include drug delivery system materials, molding aids, packaging materials, artificial blood vessels, hemodialysis membranes, catheters, guard wires, contact lenses, blood filters, blood storage packs, endoscopes, artificial organs, biotechnology A chip, a cell culture sheet, and a sugar chain synthesizer can be mentioned, but there are no particular limitations on the medical instrument.
  • the silane coupling agent (surface treatment agent) composition containing the silylalkyl phosphoramidade compound of the present invention includes, for example, water and a polar organic solvent such as methanol, ethanol, propylene glycol monomethyl ether and the like on the silylalkylphosphoramidade compound. In addition, it is prepared by diluting to 0.001 to 20% by mass.
  • a polar organic solvent such as methanol, ethanol, propylene glycol monomethyl ether and the like on the silylalkylphosphoramidade compound.
  • an organic acid may be further added. Examples of the organic acid include acetic acid, formic acid, lactic acid, and oxalic acid.
  • the surface treatment of the substrate with the silane coupling agent composition of the present invention is not particularly limited, and for example, the substrate can be treated by dipping, coating (spin coating, spray coating, etc.) or vapor deposition.
  • silane coupling is performed through a step of applying and baking the silane coupling agent composition of the present invention on a substrate, a step of washing the substrate with a polar solvent, and a step of drying the substrate. Immobilize the agent.
  • the silane coupling agent is immobilized through a step of immersing the substrate in the silane coupling agent composition, a step of washing the substrate with a polar solvent, and a step of drying the substrate.
  • the polar solvent used in the washing step for example, water or a polar organic solvent contained in the silane coupling agent composition can be used.
  • 2-Chloro-2-oxo-1,3,2-dioxaphospholane (3.37 g) was dissolved in tetrahydrofuran (20 mL), cooled to 0 ° C., triethylamine (2.88 g, 1.2 molar equivalent) was added, and the mixture was stirred. . 3.82 g (1.0 molar equivalent) of (3-aminopropyl) dimethylethoxysilane was added dropwise, and the temperature was gradually raised to 25 ° C. After stirring overnight, the insoluble material was filtered under reduced pressure.
  • Example 4 Treatment of 2- (trimethylammonio) ethyl (3- (ethoxydimethylsilyl) propyl) phosphoramidate on a glass substrate
  • 2- (Trimethylammonio) ethyl (3- (ethoxy) synthesized in Example 2 10 mg of dimethylsilyl) propyl) phosphoramidate is dissolved in 1 mL of 70% ethanol aqueous solution, and 0.1 mL is dropped on a glass substrate (a square with a side of 2 cm) attached to a spin coater, and 300 rpm for 5 seconds, followed by 2500 rpm, 25 Spin coating was performed for 2 seconds. Then, it heat-processed for 60 second with a 100 degreeC hotplate, and was fixed.
  • a similar treatment was performed on a solution of 30 mg of 2- (trimethylammonio) ethyl (3- (ethoxydimethylsilyl) propyl) phosphoramidate in 1 mL of 70% ethanol aqueous
  • Example 5 Treatment of 2- (trimethylammonio) ethyl (3- (triethoxysilyl) propyl) phosphoramidate on a glass substrate
  • 2- (trimethylammonio) ethyl (3- (tri 30 mg of ethoxysilyl) propyl) phosphoramidate is dissolved in 1 mL of 70% ethanol aqueous solution, and 0.1 mL is dropped on a glass substrate (a square with a side of 2 cm) mounted on a spin coater, and 300 rpm for 5 seconds, followed by 2500 rpm, 25 Spin coating was performed for 2 seconds. Furthermore, it was fixed by heat treatment for 60 seconds on a 100 ° C. hot plate.
  • Comparative Example 1 Treatment of MPC polymer on glass substrate 15 mg of MPC (2-methacryloyloxyphosphorylcholine) polymer (PUREBRIGHT MB, registered by NOF Corporation) was dissolved in 0.5 mL of 70% aqueous ethanol solution, and 0.1 mL was spinned. It was dropped on a glass substrate (a square having a side of 2 cm) mounted on the coater, and spin coating was performed at 300 rpm for 5 seconds followed by 2500 rpm for 25 seconds. Furthermore, it was fixed by heat treatment for 60 seconds on a 100 ° C. hot plate.
  • MPC (2-methacryloyloxyphosphorylcholine) polymer PUREBRIGHT MB, registered by NOF Corporation
  • Example 6 Confirmation of Cell Adhesion Inhibition Action on Glass Substrate Treated with 2- (Trimethylammonio) ethyl (3- (ethoxydimethylsilyl) propyl) Phosphoramidate HEK293 cells (human kidney-derived cell line Human Embryonic Kidney ( 2 ⁇ 10 6 cells / ml) was pre-cultured in a Dulbecco's Modified Eagle Medium (D-MEM) medium at 37 ° C. for 2 days, and 3 mL seeded in a 12-well plate containing a glass substrate treated by the method of Example 4 Then, 0.8 mL of D-MEM was added and cultured for 1 day at 37 ° C.
  • D-MEM Dulbecco's Modified Eagle Medium
  • the glass substrate was then transferred to another 12-well plate and 1 mL phosphate buffer (Dulbecco phosphate buffered saline, Dulbecco's Phosphate Buffered Sali
  • phosphate buffer Dulbecco phosphate buffered saline, Dulbecco's Phosphate Buffered Sali
  • the cells were detached from the glass substrate by adding 0.3 mL of a 0.1% trypsin solution (manufactured by SIGMA), to which 3 mL of D-MEM was added, and WST was further added.
  • 0.3 mL of -8 Korean (registered trademark) was added to each well and incubated at 37 ° C. for 4 hours, and then 0.11 mL was transferred to a 96-well plate and the absorbance was measured.
  • 2- (trimethylammonio) ethyl (3- (ethoxydimethylsilyl) propyl) phosphoramidate is obtained by treating 0.1 mL of 10 mg / mL and 30 mg / mL solutions on a glass substrate. The adhesion of HEK cells to the glass substrate was suppressed.
  • Example 7 Measurement of living cells attached to a treated glass substrate.
  • HEK293 cells human kidney-derived cell line Human Embroidic Kidney (2 ⁇ 10 6 cells / ml) pre-cultured in Dulbecco's Modified Eagle Medium (D-MEM) medium at 37 ° C. for 2 days were treated in Example 4.
  • D-MEM Dulbecco's Modified Eagle Medium
  • these glass substrates were transferred to another 6-well plate and washed with 1 mL of phosphate buffer (Dulbecco's phosphate buffered saline, Dulbecco's Phosphate Buffered Saline), respectively, and 1 mL of D-MEM was then removed.
  • phosphate buffer Dulbecco's phosphate buffered saline, Dulbecco's Phosphate Buffered Saline
  • WST-8 Wooda Chemical
  • 2- (trimethylammonio) ethyl (3- (ethoxydimethylsilyl) propyl) phosphoramidate and 2- (trimethylammonio) ethyl (3- (triethoxysilyl) propyl) phosphoramidate are By treating 0.1 mL of a 30 mg / mL solution on a glass substrate, HEK cell adhesion to the glass substrate was suppressed. This inhibitory effect was equivalent to or better than that of MPC polymer.

Abstract

[Problem] To provide a novel silane coupling agent. [Solution] A silylalkyl phosphoramidate compound represented by formula (1), and a silane coupling agent composition which contains the compound. (In the formula, each of R1, R2 and R3 independently represents an alkyl group having 1-5 carbon atoms; each of R4 and R5 independently represents an alkyl group having 1-5 carbon atoms; each of n and p independently represents an integer of 1-5; and q represents an integer of 1-3.)

Description

シリルアルキルホスホラミダード化合物を含むシランカップリング剤Silane coupling agents containing silylalkyl phosphoramidade compounds
　本発明は、シリコン、ガラス等の無機物質、又はポリエチレン等の樹脂に対して表面処理を行うことにより、タンパク質、細胞、血小板等の生体物質の付着を抑制可能なシリルアルキルホスホラミダード化合物を含むシランカップリング剤に関する。 The present invention includes a silylalkyl phosphoramide compound capable of suppressing the adhesion of biological materials such as proteins, cells, and platelets by performing a surface treatment on an inorganic material such as silicon or glass, or a resin such as polyethylene. The present invention relates to a silane coupling agent.
　生体系は、人工材料など外的なものと接触した場合、当該人工材料を異物として認識するため、時間の経過に伴い血栓形成、免疫反応、炎症反応など様々な異物反応が引き起こされる。そのため、人工臓器などの医療用器具を用いる場合、ヘパリンなどの抗血液凝固剤、免疫抑制剤のような薬剤を併用しなければならない。 When a biological system comes into contact with an external material such as an artificial material, the artificial material is recognized as a foreign material, and thus various foreign material reactions such as thrombus formation, immune reaction, and inflammatory reaction are caused over time. Therefore, when a medical instrument such as an artificial organ is used, an anticoagulant such as heparin or a drug such as an immunosuppressant must be used in combination.
　ところが、上記抗血液凝固剤等を使用した場合には、肝臓障害、アレルギー反応などの様々な副反応を生じるおそれがある。 However, when the above anticoagulant or the like is used, various side reactions such as liver damage and allergic reaction may occur.
　そこで、これらの課題を解決するために、生体膜と同じ両性型のリン脂質であるホスホリルコリンを高分子鎖の側鎖に有するポリ（２－メタクリロイルオキシエチルホスホリルコリン）（以下、ＭＰＣポリマーという）を用いた医療用材料が提案されている（例えば、特許文献１及び特許文献２参照）。 Therefore, in order to solve these problems, poly (2-methacryloyloxyethyl phosphorylcholine) (hereinafter referred to as MPC polymer) having phosphorylcholine, which is the same amphoteric phospholipid as the biological membrane, in the side chain of the polymer chain is used Medical materials that have been proposed have been proposed (see, for example, Patent Document 1 and Patent Document 2).
　また、Ｎ－メタクリロイルオキシエチル－Ｎ，Ｎ－ジメチルアンモニウム－α－Ｎ－メチルカルボキシベタインの高分子量体（以下、ＣＭＢポリマーという）を用いた医療用材料が提案されている（例えば、特許文献３、特許文献４及び特許文献５参照）。 In addition, a medical material using a high molecular weight N-methacryloyloxyethyl-N, N-dimethylammonium-α-N-methylcarboxybetaine (hereinafter referred to as CMB polymer) has been proposed (for example, Patent Document 3). Patent Document 4 and Patent Document 5).
　ＭＰＣポリマー又はＣＭＢポリマーを材料表面に被覆することで、抗血液凝固剤を使用しない場合でも血液凝固を抑えることが可能となる。
　しかしながら、ＭＰＣポリマー及びＣＭＢポリマーの合成、さらに当該ポリマーを含む溶液を作製するまでの工程に手間がかかるという問題がある。 By coating the MPC polymer or CMB polymer on the surface of the material, it is possible to suppress blood coagulation even when no anticoagulant is used.
However, there is a problem that it takes time to synthesize the MPC polymer and the CMB polymer and further to prepare a solution containing the polymer.
特開昭５４－６３０２５号公報JP 54-63025 A 特開２０００－２７９５１２号公報JP 2000-279512 A 特開２００７－１３０１９４号公報JP 2007-130194 A 特開２００８－２７４１５１号公報JP 2008-274151 A 特開２０１０－５７７４５号公報JP 2010-57745 A
　本発明の課題は、製造が簡便な非ポリマーである化合物であり、該化合物が溶解した溶液を基材に塗布又は該溶液に基材を浸漬させることにより、直接、基材の表面に該化合物を強固に被覆することが可能であり、しかも、従来のＭＰＣポリマー及びＣＭＢポリマーと同等の生体物質の付着抑制効果を示す化合物を提供することにある。 An object of the present invention is a compound that is a non-polymer that is easy to produce, and the compound is directly applied to the surface of a substrate by applying the solution in which the compound is dissolved to the substrate or immersing the substrate in the solution. It is another object of the present invention to provide a compound capable of firmly covering the surface of the body and exhibiting the same biological substance adhesion inhibitory effect as the conventional MPC polymer and CMB polymer.
　本発明者らは、上記課題を解決するために鋭意検討した結果、ある種のシリルアルキルホスホラミダード化合物がシランカップリング剤として有用であり、当該化合物を含むシランカップリング剤がタンパク質、細胞、血小板等の生体物質の付着を抑制する効果を有することを見出した。
　すなわち、本発明は、
１．下記式（１）： As a result of intensive studies to solve the above problems, the present inventors have found that certain silylalkyl phosphoramidade compounds are useful as silane coupling agents, and silane coupling agents containing such compounds are proteins, cells, It has been found that it has an effect of suppressing adhesion of biological substances such as platelets.
That is, the present invention
1. Following formula (1):
Figure JPOXMLDOC01-appb-C000002
Figure JPOXMLDOC01-appb-C000002
（式中、Ｒ１、Ｒ２及びＲ３はそれぞれ独立に炭素原子数１乃至５のアルキル基を表し、Ｒ４及びＲ５はそれぞれ独立に炭素原子数１乃至５のアルキル基を表し、ｎ及びｐはそれぞれ独立に１乃至５の整数を表し、ｑは１乃至３の整数を表す。）
で表されるシリルアルキルホスホラミダード化合物、
２．上記１に記載のシリルアルキルホスホラミダード化合物を含むシランカップリング剤組成物、
３．極性有機溶媒がメタノール、エタノールおよびプロピレングリコールモノメチルエーテルからなる群より選ばれる少なくとも１種である上記２に記載のシランカップリング剤組成物、
４．上記２または３に記載のシランカップリング剤組成物を基材上に塗布しベークする工程、その後極性溶媒で前記基材を洗浄する工程、及び前記基材を乾燥させる工程を含むシランカップリング剤の固定化方法、
５．基材を上記２または３に記載のシランカップリング剤組成物中に浸漬する工程、その後極性溶媒で前記基材を洗浄する工程、及び前記基材を乾燥させる工程を含むシランカップリング剤の固定化方法、
６．上記４または５に記載の方法でシランカップリング剤を固定化した基材、
７．上記４または５に記載の方法でシランカップリング剤を固定化した医療用器具、
である。 (Wherein R 1 , R 2 and R 3 each independently represents an alkyl group having 1 to 5 carbon atoms, R 4 and R 5 each independently represents an alkyl group having 1 to 5 carbon atoms, n And p each independently represents an integer of 1 to 5, and q represents an integer of 1 to 3.
A silylalkyl phosphoramidade compound represented by:
2. A silane coupling agent composition comprising the silylalkyl phosphoramidade compound according to 1 above,
3. The silane coupling agent composition according to 2 above, wherein the polar organic solvent is at least one selected from the group consisting of methanol, ethanol and propylene glycol monomethyl ether,
4). A silane coupling agent comprising a step of applying and baking the silane coupling agent composition described in 2 or 3 above on a substrate, a step of washing the substrate with a polar solvent, and a step of drying the substrate. Immobilization method,
5. Immobilization of a silane coupling agent comprising a step of immersing a substrate in the silane coupling agent composition described in 2 or 3 above, a step of washing the substrate with a polar solvent, and a step of drying the substrate. Conversion method,
6). A substrate on which a silane coupling agent is immobilized by the method according to 4 or 5 above,
7). A medical instrument in which a silane coupling agent is immobilized by the method according to 4 or 5 above,
It is.
　本発明のシリルアルキルホスホラミダード化合物を含むシランカップリング剤（表面処理剤）組成物を用いることにより、シリコン、ガラス等の無機物質又はポリエチレン等の樹脂（以下、基材という）の表面処理が可能で、その表面は、一分子内に安定な正負両電荷を持つシリルアルキルホスホラミダード化合物で被覆される。そのため、ＭＰＣポリマー、ＣＭＢポリマーのようなポリマーを用いることなく、上記無機物質又は樹脂表面にタンパク質、細胞、血小板等の生体物質の付着を抑制することができる。 By using the silane coupling agent (surface treatment agent) composition containing the silylalkyl phosphoramidade compound of the present invention, surface treatment of an inorganic substance such as silicon or glass or a resin such as polyethylene (hereinafter referred to as a base material) is possible. It is possible that the surface is coated with a silylalkyl phosphoramidade compound having both stable positive and negative charges in one molecule. Therefore, adhesion of biological substances such as proteins, cells, and platelets to the inorganic substance or resin surface can be suppressed without using a polymer such as MPC polymer or CMB polymer.
図１は、２－（トリメチルアンモニオ）エチル（３－（エトキシジメチルシリル）プロピル）ホスホラミダートのガラス基板への細胞付着抑制作用を示す図である。細胞フリーは、２－（トリメチルアンモニオ）エチル（３－（エトキシジメチルシリル）プロピル）ホスホラミダート（３０ｍｇ／ｍｌ）で処理したガラス基板を表し、コントロールは、本発明のシランカップリング剤での処理をしていない場合の結果を表す。FIG. 1 is a diagram showing the cell adhesion inhibitory action of 2- (trimethylammonio) ethyl (3- (ethoxydimethylsilyl) propyl) phosphoramidate on a glass substrate. Cell free represents a glass substrate treated with 2- (trimethylammonio) ethyl (3- (ethoxydimethylsilyl) propyl) phosphoramidate (30 mg / ml), and the control was controlled with the silane coupling agent of the present invention. Represents the result when no processing is performed. 図２は、シリルアルキルホスホラミダード化合物のガラス基板に付着した生細胞測定の結果を示す図である。コントロールは、本発明のシランカップリング剤での処理をしていない場合の結果を表す。FIG. 2 is a diagram showing the results of measurement of living cells attached to a glass substrate of a silylalkyl phosphoramidade compound. Control represents the result when the treatment with the silane coupling agent of the present invention is not performed.
　本明細書における「ｎ」はノルマルを、「ｓ」はセカンダリーを、「ｔ」はターシャリーを表す。
　前記式（１）で表される本発明のシリルアルキルホスホラミダード化合物は、式（Ｉ）で表されるアミノアルキルシランに、０．８乃至１．２当量、好ましくは１当量の式（ＩＩ）で表される環状リン化合物を、塩基存在下、２０乃至８０℃で反応させて式（ＩＩＩ）で表される化合物を得た後、式（ＩＩＩ）で表される化合物に対して０．８乃至１．２当量、好ましくは１当量の式（ＩＶ）で表される三級アミンを０乃至５０℃で反応させることにより得ることができる。 In this specification, “n” represents normal, “s” represents secondary, and “t” represents tertiary.
The silylalkyl phosphoramidade compound of the present invention represented by the formula (1) is 0.8 to 1.2 equivalents, preferably 1 equivalent of the formula (II) to the aminoalkylsilane represented by the formula (I). ) Is reacted at 20 to 80 ° C. in the presence of a base to obtain a compound represented by the formula (III). It can be obtained by reacting 8 to 1.2 equivalents, preferably 1 equivalent of a tertiary amine represented by the formula (IV) at 0 to 50 ° C.
Figure JPOXMLDOC01-appb-C000003
Figure JPOXMLDOC01-appb-C000003
　式（Ｉ）で表されるアミノアルキルシランと式（ＩＩ）で表される環状リン化合物との反応で用いられる塩基としては、トリエチルアミン等の有機塩基が好ましい。 As the base used in the reaction between the aminoalkylsilane represented by the formula (I) and the cyclic phosphorus compound represented by the formula (II), an organic base such as triethylamine is preferable.
　本発明の式（１）で表されるシリルアルキルホスホラミダード化合物において、Ｒ１、Ｒ２及びＲ３としてはメチル基、エチル基、ｎ－プロピル基、イソプロピル基、シクロプロピル基、ｎ－ブチル基、イソブチル基、ｓ－ブチル基、ｔ－ブチル基、シクロブチル基、ｎ－ペンチル基、シクロペンチル基が挙げられるが、特に、メチル基およびエチル基が好ましい。 In the silylalkylphosphoramidade compound represented by the formula (1) of the present invention, R 1 , R 2 and R 3 are methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl. Group, isobutyl group, s-butyl group, t-butyl group, cyclobutyl group, n-pentyl group, and cyclopentyl group, and methyl group and ethyl group are particularly preferable.
　本発明の式（１）で表されるシリルアルキルホスホラミダード化合物において、Ｒ４としてはメチル基、エチル基、ｎ－プロピル基、イソプロピル基、シクロプロピル基、ｎ－ブチル基、イソブチル基、ｓ－ブチル基、ｔ－ブチル基、シクロブチル基、ｎ－ペンチル基、シクロペンチル基が挙げられるが、特に、メチル基およびエチル基が好ましい。 In the silylalkylphosphoramidade compound represented by the formula (1) of the present invention, R 4 is methyl group, ethyl group, n-propyl group, isopropyl group, cyclopropyl group, n-butyl group, isobutyl group, s -Butyl group, t-butyl group, cyclobutyl group, n-pentyl group, and cyclopentyl group can be mentioned, and methyl group and ethyl group are particularly preferable.
　本発明の式（１）で表されるシリルアルキルホスホラミダード化合物において、Ｒ５としてはメチル基、エチル基、ｎ－プロピル基、イソプロピル基、シクロプロピル基、ｎ－ブチル基、イソブチル基、ｓ－ブチル基、ｔ－ブチル基、シクロブチル基、ｎ－ペンチル基、シクロペンチル基が挙げられるが、特に、メチル基およびエチル基が好ましい。 In the silylalkylphosphoramidade compound represented by the formula (1) of the present invention, R 5 represents methyl group, ethyl group, n-propyl group, isopropyl group, cyclopropyl group, n-butyl group, isobutyl group, s -Butyl group, t-butyl group, cyclobutyl group, n-pentyl group, and cyclopentyl group can be mentioned, and methyl group and ethyl group are particularly preferable.
　本発明の式（１）で表されるシリルアルキルホスホラミダード化合物において、ｎとしては１乃至５の整数が挙げられるが、特に、２または３が好ましい。ｐとしては１乃至５の整数が挙げられるが、特に、２または３が好ましい。ｑとしては１乃至３の整数が挙げられる。 In the silylalkyl phosphoramidade compound represented by the formula (1) of the present invention, n may be an integer of 1 to 5, and 2 or 3 is particularly preferable. p may be an integer of 1 to 5, but 2 or 3 is particularly preferable. q is an integer of 1 to 3.
　本発明のシリルアルキルホスホラミダード化合物を基材である無機物質の表面処理に用いる場合、その無機物質には特に制限がない。例えば、シリコン、銅、鉄、アルミニウム、亜鉛またはそれらの合金、ガラス、シリカ、酸化アルミニウム、水酸化アルミニウム、酸化マグネシウムが挙げられる。 When the silylalkyl phosphoramidade compound of the present invention is used for surface treatment of an inorganic substance as a base material, the inorganic substance is not particularly limited. For example, silicon, copper, iron, aluminum, zinc or an alloy thereof, glass, silica, aluminum oxide, aluminum hydroxide, and magnesium oxide can be given.
　本発明のシリルアルキルホスホラミダード化合物を基材である樹脂の表面処理に用いる場合、その樹脂には特に制限がない。例えば、ポリエチレン、ポリプロピレン、ポリスチレン、ポリ塩化ビニル、ナイロン、ポリウレタン、ポリウレア、ポリ乳酸、ポリグリコール酸、ポリビニルアルコール、ポリ酢酸ビニル、ポリ（メタ）アクリル酸、ポリ（メタ）アクリル酸誘導体、ポリアクリロニトリル、ポリ（メタ）アクリルアミド、ポリ（メタ）アクリルアミド誘導体、ポリスルホン、ポリカーボネート、セルロース、セルロース誘導体が挙げられる。 When the silylalkyl phosphoramidade compound of the present invention is used for the surface treatment of a resin as a base material, the resin is not particularly limited. For example, polyethylene, polypropylene, polystyrene, polyvinyl chloride, nylon, polyurethane, polyurea, polylactic acid, polyglycolic acid, polyvinyl alcohol, polyvinyl acetate, poly (meth) acrylic acid, poly (meth) acrylic acid derivatives, polyacrylonitrile, Examples include poly (meth) acrylamide, poly (meth) acrylamide derivatives, polysulfone, polycarbonate, cellulose, and cellulose derivatives.
　本発明のシリルアルキルホスホラミダード化合物は、医薬品、医薬部外品、医療用器具等の表面処理が可能である。前記医療用器具の例として、ドラッグデリバリーシステム材、成形補助材、包装材、人工血管、血液透析膜、カテーテル、ガードワイヤー、コンタクトレンズ、血液フィルター、血液保存パック、内視鏡、人工臓器、バイオチップ、細胞培養シート、糖鎖合成機器が挙げられるが、その医療用器具には特に制限がない。 The silylalkyl phosphoramidade compound of the present invention can be used for surface treatment of pharmaceuticals, quasi drugs, medical instruments and the like. Examples of medical devices include drug delivery system materials, molding aids, packaging materials, artificial blood vessels, hemodialysis membranes, catheters, guard wires, contact lenses, blood filters, blood storage packs, endoscopes, artificial organs, biotechnology A chip, a cell culture sheet, and a sugar chain synthesizer can be mentioned, but there are no particular limitations on the medical instrument.
　本発明のシリルアルキルホスホラミダード化合物を含むシランカップリング剤（表面処理剤）組成物は、例えば、当該シリルアルキルホスホラミダード化合物に水及びメタノール、エタノール、プロピレングリコールモノメチルエーテル等の極性有機溶媒を加え０．００１乃至２０質量％になるように希釈して、調製される。前記シランカップリング剤組成物のｐＨを調整するために、さらに有機酸を添加してもよい。前記有機酸としては、例えば、酢酸、蟻酸、乳酸、シュウ酸が挙げられる。 The silane coupling agent (surface treatment agent) composition containing the silylalkyl phosphoramidade compound of the present invention includes, for example, water and a polar organic solvent such as methanol, ethanol, propylene glycol monomethyl ether and the like on the silylalkylphosphoramidade compound. In addition, it is prepared by diluting to 0.001 to 20% by mass. In order to adjust the pH of the silane coupling agent composition, an organic acid may be further added. Examples of the organic acid include acetic acid, formic acid, lactic acid, and oxalic acid.
　前記基材に、本発明のシランカップリング剤組成物で表面処理する方法は特に限定されず、例えば、浸漬、塗布（スピンコート、スプレーコートなど）又は蒸着によって処理できる。 The surface treatment of the substrate with the silane coupling agent composition of the present invention is not particularly limited, and for example, the substrate can be treated by dipping, coating (spin coating, spray coating, etc.) or vapor deposition.
　具体的には、本発明のシランカップリング剤組成物を基材上に塗布しベークする工程、その後極性溶媒で前記基材を洗浄する工程、及び前記基材を乾燥させる工程を経てシランカップリング剤を固定化させる。あるいは、基材を前記シランカップリング剤組成物中に浸漬する工程、その後極性溶媒で前記基材を洗浄する工程、及び前記基材を乾燥させる工程を経てシランカップリング剤を固定化させる。洗浄工程に使用する極性溶媒としては、例えば、前記シランカップリング剤組成物に含まれる水又は極性有機溶媒を使用することができる。 Specifically, silane coupling is performed through a step of applying and baking the silane coupling agent composition of the present invention on a substrate, a step of washing the substrate with a polar solvent, and a step of drying the substrate. Immobilize the agent. Alternatively, the silane coupling agent is immobilized through a step of immersing the substrate in the silane coupling agent composition, a step of washing the substrate with a polar solvent, and a step of drying the substrate. As the polar solvent used in the washing step, for example, water or a polar organic solvent contained in the silane coupling agent composition can be used.
　以下に、実施例を挙げて本発明をより具体的に説明するが、本発明はこれらに限定されるものではない。
　なお、１Ｈ－ＮＭＲおよび吸光度は以下の機器および条件で測定した。また、ＮＭＲは核磁気共鳴スペクトルを表す。
［１］１Ｈ－ＮＭＲ
　機種：ＪＮＭ－ＥＣＰ３００　（ＪＥＯＬ製）（３００ＭＨｚ）
　測定溶媒：重クロロホルムまたは重水
［２］吸光度測定
　機種：マイクロプレートリーダーＳＰＥＣＴＲＡ　ｍａｘ（日本モレキュラーデバイス社製）
　波長：４５０ｎｍ
　また、実施例の１Ｈ－ＮＭＲのケミカルシフト値における記号は、下記の意味を表す。
ｓ：シングレット、ｄ：ダブレット、ｔ：トリプレット、ｍ：マルチプレット。 Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to these examples.
1 H-NMR and absorbance were measured with the following equipment and conditions. NMR represents a nuclear magnetic resonance spectrum.
[1] 1 H-NMR
Model: JNM-ECP300 (manufactured by JEOL) (300MHz)
Measurement solvent: deuterated chloroform or deuterated water [2] Absorbance measurement Model: Microplate reader SPECTRA max (manufactured by Nihon Molecular Devices)
Wavelength: 450nm
Further, the symbols in the chemical shift values of 1 H-NMR in the examples have the following meanings.
s: singlet, d: doublet, t: triplet, m: multiplet.
実施例１：２－（（３－（エトキシジメチルシリル）プロピル）アミノ）－２－オキソ－１，３，２－ジオキサホスホランの合成 Example 1: Synthesis of 2-((3- (ethoxydimethylsilyl) propyl) amino) -2-oxo-1,3,2-dioxaphospholane
Figure JPOXMLDOC01-appb-C000004
Figure JPOXMLDOC01-appb-C000004
　２－クロロ－２－オキソ－１，３，２－ジオキサホスホラン３．３７ｇをテトラヒドロフラン２０ｍＬに溶解させて０℃に冷却し、トリエチルアミン２．８８ｇ（１．２モル当量）を加えて撹拌した。（３－アミノプロピル）ジメチルエトキシシラン３．８２ｇ（１．０モル当量）を滴下し、徐々に２５℃に昇温させた。一晩撹拌した後、不溶物を減圧ろ過した。ろ液を濃縮することで、黄色油状物として４．８９ｇ（収率７７％）の２－（（３－（エトキシジメチルシリル）プロピル）アミノ）－２－オキソ－１，３，２－ジオキサホスホランを得た。
　１Ｈ　ＮＭＲ（ｐｐｍ、３００ＭＨｚ、重クロロホルム）：σ＝４．３（ｍ、１Ｈ）、４．２（ｍ、１Ｈ）、４．１（ｍ、１Ｈ）、３．６（ｍ、１Ｈ）、３．５（ｄｄ、２Ｈ）、３．０（ｄｄ、１Ｈ）、２．８（ｍ、２Ｈ）、１．４（ｍ、１Ｈ）、１．１（ｔ、３Ｈ）、０．５（ｍ、２Ｈ）、０．０（ｓ、６Ｈ）。 2-Chloro-2-oxo-1,3,2-dioxaphospholane (3.37 g) was dissolved in tetrahydrofuran (20 mL), cooled to 0 ° C., triethylamine (2.88 g, 1.2 molar equivalent) was added, and the mixture was stirred. . 3.82 g (1.0 molar equivalent) of (3-aminopropyl) dimethylethoxysilane was added dropwise, and the temperature was gradually raised to 25 ° C. After stirring overnight, the insoluble material was filtered under reduced pressure. The filtrate was concentrated to give 4.89 g (yield 77%) of 2-((3- (ethoxydimethylsilyl) propyl) amino) -2-oxo-1,3,2-dioxa as a yellow oil. A phosphorane was obtained.
1 H NMR (ppm, 300 MHz, deuterated chloroform): σ = 4.3 (m, 1H), 4.2 (m, 1H), 4.1 (m, 1H), 3.6 (m, 1H), 3.5 (dd, 2H), 3.0 (dd, 1H), 2.8 (m, 2H), 1.4 (m, 1H), 1.1 (t, 3H), 0.5 (m 2H), 0.0 (s, 6H).
実施例２：２－（トリメチルアンモニオ）エチル（３－（エトキシジメチルシリル）プロピル）ホスホラミダートの合成 Example 2: Synthesis of 2- (trimethylammonio) ethyl (3- (ethoxydimethylsilyl) propyl) phosphoramidate
Figure JPOXMLDOC01-appb-C000005
Figure JPOXMLDOC01-appb-C000005
　得られた２－（（３－（エトキシジメチルシリル）プロピル）アミノ）－２－オキソー１，３，２－ジオキサホスホラン２．００ｇをアセトニトリル５ｍＬに溶解させ、トリメチルアミン９１０ｍｇのテトラヒドロフラン溶液５ｍＬを加えた。溶液を４５℃で２４時間撹拌し、析出した結晶を減圧ろ過により回収することで、無色結晶として２－（トリメチルアンモニオ）エチル（３－（エトキシジメチルシリル）プロピル）ホスホラミダートを３５０ｍｇ（収率１４％）得た。
　１Ｈ　ＮＭＲ（ｐｐｍ、３００ＭＨｚ、重水）：σ＝４．１（ｄｄ、４Ｈ）、３．５（ｄｄ、２Ｈ）、３．１（ｍ、１Ｈ）、２．９（ｍ、１Ｈ）、２．８（ｓ、９Ｈ）、１．６（ｍ、１Ｈ）、１．４（ｍ、１Ｈ）、１．０（ｔ、３Ｈ）、０．５（ｍ、２Ｈ）、０．０（ｓ、６Ｈ）。 2.00 g of the obtained 2-((3- (ethoxydimethylsilyl) propyl) amino) -2-oxo-1,3,2-dioxaphosphorane was dissolved in 5 mL of acetonitrile, and 5 mL of a tetrahydrofuran solution of 910 mg of trimethylamine was added. It was. The solution was stirred at 45 ° C. for 24 hours, and the precipitated crystals were collected by filtration under reduced pressure to give 350 mg (yield) of 2- (trimethylammonio) ethyl (3- (ethoxydimethylsilyl) propyl) phosphoramidate as colorless crystals. 14%).
1 H NMR (ppm, 300 MHz, heavy water): σ = 4.1 (dd, 4H), 3.5 (dd, 2H), 3.1 (m, 1H), 2.9 (m, 1H), 2 .8 (s, 9H), 1.6 (m, 1H), 1.4 (m, 1H), 1.0 (t, 3H), 0.5 (m, 2H), 0.0 (s, 6H).
実施例３：２－（トリメチルアンモニオ）エチル（３－（トリエトキシシリル）プロピル）ホスホラミダートの合成 Example 3: Synthesis of 2- (trimethylammonio) ethyl (3- (triethoxysilyl) propyl) phosphoramidate
Figure JPOXMLDOC01-appb-C000006
Figure JPOXMLDOC01-appb-C000006
　３－アミノプロピルトリエトキシシラン１．００ｇをテトラヒドロフラン５ｍＬに溶解させて５℃に冷却し、トリエチルアミン０．５０ｇ（１．１モル当量）を加えて撹拌した。２－クロロ－２－オキソ－１，３，２－ジオキサホスホラン０．６４ｇ（１．０モル当量）を滴下し、徐々に２０℃に昇温させた。一晩撹拌した後、不溶物をろ過除去した。ろ液を濃縮することで無色結晶として１．５６ｇ（収率６８％）の２－（３－トリエトキシシリル）プロピルアミノ）－２－オキソ－１，３，２－ジオキサホスホランを得た。
　得られた２－（３－トリエトキシシリル）プロピルアミノ）－２－オキソ－１，３，２－ジオキサホスホラン１．５６ｇをアセトニトリル５ｍＬに溶解させ、トリメチルアミン８２０ｍｇのテトラヒドロフラン溶液５ｍＬを加えた。溶液を４５℃で４８時間撹拌し、析出した結晶を減圧ろ過により回収することで、無色結晶として２－（トリメチルアンモニオ）エチル（３－（トリエトキシシリル）プロピル）ホスホラミダートを３００ｍｇ（収率１６％）得た。
　１Ｈ　ＮＭＲ（ｐｐｍ、３００ＭＨｚ、重水）：σ＝４．１（ｄｄ、４Ｈ）、３．５（ｄｄ、６Ｈ）、３．１（ｍ、１Ｈ）、２．９（ｍ、１Ｈ）、２．８（ｓ、９Ｈ）、１．６（ｍ、１Ｈ）、１．４（ｍ、１Ｈ）、１．０（ｔ、９Ｈ）、０．５（ｍ、２Ｈ）。 1.00 g of 3-aminopropyltriethoxysilane was dissolved in 5 mL of tetrahydrofuran, cooled to 5 ° C., 0.50 g (1.1 molar equivalent) of triethylamine was added and stirred. 0.64 g (1.0 molar equivalent) of 2-chloro-2-oxo-1,3,2-dioxaphosphorane was added dropwise, and the temperature was gradually raised to 20 ° C. After stirring overnight, the insoluble material was removed by filtration. The filtrate was concentrated to obtain 1.56 g (68% yield) of 2- (3-triethoxysilyl) propylamino) -2-oxo-1,3,2-dioxaphospholane as colorless crystals. .
1.56 g of the obtained 2- (3-triethoxysilyl) propylamino) -2-oxo-1,3,2-dioxaphosphorane was dissolved in 5 mL of acetonitrile, and 5 mL of a tetrahydrofuran solution of 820 mg of trimethylamine was added. The solution was stirred at 45 ° C. for 48 hours, and the precipitated crystals were collected by vacuum filtration to give 300 mg (yield) of 2- (trimethylammonio) ethyl (3- (triethoxysilyl) propyl) phosphoramidate as colorless crystals. (Rate 16%).
1 H NMR (ppm, 300 MHz, heavy water): σ = 4.1 (dd, 4H), 3.5 (dd, 6H), 3.1 (m, 1H), 2.9 (m, 1H), 2 .8 (s, 9H), 1.6 (m, 1H), 1.4 (m, 1H), 1.0 (t, 9H), 0.5 (m, 2H).
実施例４：ガラス基板への２－（トリメチルアンモニオ）エチル（３－（エトキシジメチルシリル）プロピル）ホスホラミダートの処理
　実施例２において合成した２－（トリメチルアンモニオ）エチル（３－（エトキシジメチルシリル）プロピル）ホスホラミダート１０ｍｇを７０％エタノール水溶液１ｍＬに溶解し、０．１ｍＬをスピンコーターに装着したガラス基板（一辺が２ｃｍの正方形）に滴下して３００ｒｐｍ　５秒間に続いて２５００ｒｐｍ、２５秒間処理のスピンコートを行なった。その後、１００℃ホットプレートで６０秒加熱処理し、固定化させた。
　２－（トリメチルアンモニオ）エチル（３－（エトキシジメチルシリル）プロピル）ホスホラミダート３０ｍｇを７０％エタノール水溶液１ｍＬに溶解した溶液についても、同様の処理を行った。 Example 4: Treatment of 2- (trimethylammonio) ethyl (3- (ethoxydimethylsilyl) propyl) phosphoramidate on a glass substrate 2- (Trimethylammonio) ethyl (3- (ethoxy) synthesized in Example 2 10 mg of dimethylsilyl) propyl) phosphoramidate is dissolved in 1 mL of 70% ethanol aqueous solution, and 0.1 mL is dropped on a glass substrate (a square with a side of 2 cm) attached to a spin coater, and 300 rpm for 5 seconds, followed by 2500 rpm, 25 Spin coating was performed for 2 seconds. Then, it heat-processed for 60 second with a 100 degreeC hotplate, and was fixed.
A similar treatment was performed on a solution of 30 mg of 2- (trimethylammonio) ethyl (3- (ethoxydimethylsilyl) propyl) phosphoramidate in 1 mL of 70% ethanol aqueous solution.
実施例５：ガラス基板への２－（トリメチルアンモニオ）エチル（３－（トリエトキシシリル）プロピル）ホスホラミダートの処理
　実施例３において合成した２－（トリメチルアンモニオ）エチル（３－（トリエトキシシリル）プロピル）ホスホラミダート３０ｍｇを７０％エタノール水溶液１ｍＬに溶解し、０．１ｍＬをスピンコーターに装着したガラス基板（一辺が２ｃｍの正方形）に滴下して３００ｒｐｍ　５秒間に続いて２５００ｒｐｍ、２５秒間処理のスピンコートを行なった。さらに、１００℃ホットプレートで６０秒加熱処理し、固定化させた。 Example 5: Treatment of 2- (trimethylammonio) ethyl (3- (triethoxysilyl) propyl) phosphoramidate on a glass substrate 2- (trimethylammonio) ethyl (3- (tri 30 mg of ethoxysilyl) propyl) phosphoramidate is dissolved in 1 mL of 70% ethanol aqueous solution, and 0.1 mL is dropped on a glass substrate (a square with a side of 2 cm) mounted on a spin coater, and 300 rpm for 5 seconds, followed by 2500 rpm, 25 Spin coating was performed for 2 seconds. Furthermore, it was fixed by heat treatment for 60 seconds on a 100 ° C. hot plate.
比較例１：ガラス基板へのＭＰＣポリマーの処理
　ＭＰＣ（２－メタクロイルオキシホスホリルコリン）ポリマー（ＰＵＲＥＢＲＩＧＨＴ　ＭＢ　日本油脂社製　登録商標）１５ｍｇを７０％エタノール水溶液０．５ｍＬに溶解し、０．１ｍＬをスピンコーターに装着したガラス基板（一辺が２ｃｍの正方形）に滴下して３００ｒｐｍ　５秒間に続いて２５００ｒｐｍ、２５秒間処理のスピンコートを行なった。さらに、１００℃ホットプレートで６０秒加熱処理し、固定化させた。 Comparative Example 1: Treatment of MPC polymer on glass substrate 15 mg of MPC (2-methacryloyloxyphosphorylcholine) polymer (PUREBRIGHT MB, registered by NOF Corporation) was dissolved in 0.5 mL of 70% aqueous ethanol solution, and 0.1 mL was spinned. It was dropped on a glass substrate (a square having a side of 2 cm) mounted on the coater, and spin coating was performed at 300 rpm for 5 seconds followed by 2500 rpm for 25 seconds. Furthermore, it was fixed by heat treatment for 60 seconds on a 100 ° C. hot plate.
実施例６：２－（トリメチルアンモニオ）エチル（３－（エトキシジメチルシリル）プロピル）ホスホラミダート処理したガラス基板への細胞付着抑制作用の確認
　ＨＥＫ２９３細胞（ヒト腎臓由来の細胞株Ｈｕｍａｎ　Ｅｍｂｒｙｏｎｉｃ　Ｋｉｄｎｅｙ　（２×１０６ｃｅｌｌｓ／ｍｌ）　をＤｕｌｂｅｃｃｏ’ｓ　Ｍｏｄｉｆｉｅｄ　Ｅａｇｌｅ　Ｍｅｄｉｕｍ（Ｄ－ＭＥＭ）培地で３７℃で２日間前培養したものを、実施例４の方法により処理したガラス基板を入れた１２穴プレートに３ｍＬ播種し、０．８ｍＬの　Ｄ－ＭＥＭを加えて、３７℃で１日　培養した。次いで、ガラス基板を、別の１２穴プレートに移して、１ｍＬリン酸緩衝液（ダルベッコリン酸緩衝生理食塩水、Ｄｕｌｂｅｃｃｏ’ｓ　Ｐｈｏｓｐｈａｔｅ　Ｂｕｆｆｅｒｅｄ　Ｓａｌｉｎｅ）で洗浄した。次に、０．３ｍＬの０．１％トリプシン溶液（ＳＩＧＭＡ社製）を加えることにより、細胞をガラス基板から剥がした。そこへ、３ｍＬのＤ－ＭＥＭを加え、さらにＷＳＴ－８（キシダ化学　登録商標）の０．３ｍＬを各ウエルに加えて、３７℃にて４時間インキュベート後、そのうち０．１１ｍＬを９６穴プレートに移して、吸光度を測定した。 Example 6: Confirmation of Cell Adhesion Inhibition Action on Glass Substrate Treated with 2- (Trimethylammonio) ethyl (3- (ethoxydimethylsilyl) propyl) Phosphoramidate HEK293 cells (human kidney-derived cell line Human Embryonic Kidney ( 2 × 10 6 cells / ml) was pre-cultured in a Dulbecco's Modified Eagle Medium (D-MEM) medium at 37 ° C. for 2 days, and 3 mL seeded in a 12-well plate containing a glass substrate treated by the method of Example 4 Then, 0.8 mL of D-MEM was added and cultured for 1 day at 37 ° C. The glass substrate was then transferred to another 12-well plate and 1 mL phosphate buffer (Dulbecco phosphate buffered saline, Dulbecco's Phosphate Buffered Sali The cells were detached from the glass substrate by adding 0.3 mL of a 0.1% trypsin solution (manufactured by SIGMA), to which 3 mL of D-MEM was added, and WST was further added. 0.3 mL of -8 (Kishida Chemical (registered trademark)) was added to each well and incubated at 37 ° C. for 4 hours, and then 0.11 mL was transferred to a 96-well plate and the absorbance was measured.
　図１に示すとおり、２－（トリメチルアンモニオ）エチル（３－（エトキシジメチルシリル）プロピル）ホスホラミダートは、１０ｍｇ／ｍＬおよび３０ｍｇ／ｍＬ溶液の０．１ｍＬをガラス基板に処理することにより、ガラス基板へのＨＥＫ細胞の付着を抑制した。 As shown in FIG. 1, 2- (trimethylammonio) ethyl (3- (ethoxydimethylsilyl) propyl) phosphoramidate is obtained by treating 0.1 mL of 10 mg / mL and 30 mg / mL solutions on a glass substrate. The adhesion of HEK cells to the glass substrate was suppressed.
実施例７：処理したガラス基板に付着した生細胞測定。
　ＨＥＫ２９３細胞（ヒト腎臓由来の細胞株Ｈｕｍａｎ　Ｅｍｂｒｙｏｎｉｃ　Ｋｉｄｎｅｙ　（２×１０６ｃｅｌｌｓ／ｍｌ）　をＤｕｌｂｅｃｃｏ’ｓ　Ｍｏｄｉｆｉｅｄ　Ｅａｇｌｅ　Ｍｅｄｉｕｍ（Ｄ－ＭＥＭ）培地で３７℃で２日間前培養したものを、実施例４にて処理したガラス基板、実施例５にて処理したガラス基板または比較例１にて処理したガラス基板をいれた１２穴プレートに０．７ｍＬ播種し、０．８ｍＬの　Ｄ－ＭＥＭ加えて、３７℃で１日培養した。
　次いで、これらのガラス基板を、別の６穴プレートに移して、それぞれ、１ｍＬリン酸緩衝液（ダルベッコリン酸緩衝生理食塩水、Ｄｕｌｂｅｃｃｏ’ｓ　Ｐｈｏｓｐｈａｔｅ　Ｂｕｆｆｅｒｅｄ　Ｓａｌｉｎｅ）で洗浄後、１ｍＬのＤ－ＭＥＭを加え、さらにＷＳＴ－８（キシダ化学　登録商標）の０．１ｍＬを各ウエルに加えて、３７℃にて４時間インキュベート後、吸光度を測定した。 Example 7: Measurement of living cells attached to a treated glass substrate.
HEK293 cells (human kidney-derived cell line Human Embroidic Kidney (2 × 10 6 cells / ml) pre-cultured in Dulbecco's Modified Eagle Medium (D-MEM) medium at 37 ° C. for 2 days were treated in Example 4. 0.7 mL of the glass substrate treated in Example 5, or the glass substrate treated in Comparative Example 1 in a 12-well plate, 0.8 mL of D-MEM was added, Cultured on day.
Then, these glass substrates were transferred to another 6-well plate and washed with 1 mL of phosphate buffer (Dulbecco's phosphate buffered saline, Dulbecco's Phosphate Buffered Saline), respectively, and 1 mL of D-MEM was then removed. In addition, 0.1 mL of WST-8 (Kishida Chemical) was added to each well and incubated at 37 ° C. for 4 hours, and then the absorbance was measured.
　図２に示すとおり、２－（トリメチルアンモニオ）エチル（３－（エトキシジメチルシリル）プロピル）ホスホラミダートおよび２－（トリメチルアンモニオ）エチル（３－（トリエトキシシリル）プロピル）ホスホラミダートは、３０ｍｇ／ｍＬ溶液の０．１ｍＬをガラス基板に処理することにより、ガラス基板へのＨＥＫ細胞の付着を抑制した。この抑制効果は、ＭＰＣポリマーと同等かそれ以上の効果であった。 As shown in FIG. 2, 2- (trimethylammonio) ethyl (3- (ethoxydimethylsilyl) propyl) phosphoramidate and 2- (trimethylammonio) ethyl (3- (triethoxysilyl) propyl) phosphoramidate are By treating 0.1 mL of a 30 mg / mL solution on a glass substrate, HEK cell adhesion to the glass substrate was suppressed. This inhibitory effect was equivalent to or better than that of MPC polymer.

Claims (7)

  1. 　下記式（１）：
    Figure JPOXMLDOC01-appb-C000001
     （式中、Ｒ１、Ｒ２及びＲ３はそれぞれ独立に炭素原子数１乃至５のアルキル基を表し、Ｒ４及びＲ５はそれぞれ独立に炭素原子数１乃至５のアルキル基を表し、ｎ及びｐはそれぞれ独立に１乃至５の整数を表し、ｑは１乃至３の整数を表す。）
    で表されるシリルアルキルホスホラミダード化合物。     Following formula (1):
    Figure JPOXMLDOC01-appb-C000001
     (Wherein R 1 , R 2 and R 3 each independently represents an alkyl group having 1 to 5 carbon atoms, R 4 and R 5 each independently represents an alkyl group having 1 to 5 carbon atoms, n And p each independently represents an integer of 1 to 5, and q represents an integer of 1 to 3.
    The silyl alkyl phosphoramidade compound represented by these.
  2. 　請求項１に記載のシリルアルキルホスホラミダード化合物、極性有機溶媒及び水を含むシランカップリング剤組成物。 A silane coupling agent composition comprising the silylalkyl phosphoramidade compound according to claim 1, a polar organic solvent, and water.
  3. 　前記極性有機溶媒がメタノール、エタノールおよびプロピレングリコールモノメチルエーテルからなる群より選ばれる少なくとも１種である請求項２記載のシランカップリング剤組成物。 The silane coupling agent composition according to claim 2, wherein the polar organic solvent is at least one selected from the group consisting of methanol, ethanol and propylene glycol monomethyl ether.
  4. 　請求項２または請求項３に記載のシランカップリング剤組成物を基材上に塗布しベークする工程、その後極性溶媒で前記基材を洗浄する工程、及び前記基材を乾燥させる工程を含むシランカップリング剤の固定化方法。 A silane comprising a step of applying and baking the silane coupling agent composition according to claim 2 or 3 on a substrate, a step of washing the substrate with a polar solvent, and a step of drying the substrate. Coupling agent immobilization method.
  5. 　基材を請求項２または請求項３に記載のシランカップリング剤組成物中に浸漬する工程、その後極性溶媒で前記基材を洗浄する工程、及び前記基材を乾燥させる工程を含むシランカップリング剤の固定化方法。 Silane coupling including a step of immersing a base material in the silane coupling agent composition according to claim 2 or 3, a step of washing the base material with a polar solvent, and a step of drying the base material. Immobilization method of agent.
  6. 　請求項４または請求項５に記載の方法でシランカップリング剤を固定化した基材。 The base material which fixed the silane coupling agent by the method of Claim 4 or Claim 5.
  7. 　請求項４または請求項５に記載の方法でシランカップリング剤を固定化した医療用器具。 A medical instrument in which a silane coupling agent is immobilized by the method according to claim 4 or 5.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2957563A1 (en) * 2014-06-19 2015-12-23 Pentracor GmbH Separation material comprising phosphoryl choline derivatives

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH08507293A (en) * 1993-01-28 1996-08-06 バイオコンパテイブルズ・リミテツド New material
JP2005187456A (en) * 2003-12-02 2005-07-14 Shiseido Co Ltd Phosphorylcholine group-containing compound and surface-modifying agent consisting of the same
CN101195638A (en) * 2007-12-24 2008-06-11 常州市绿空能源设备有限公司 Trinuclear phosphamide ester compounds containing tegafur and aza-silicon
JP2011236345A (en) * 2010-05-11 2011-11-24 Shiseido Co Ltd Surface modification method and surface-modified material
WO2013038901A1 (en) * 2011-09-16 2013-03-21 日産化学工業株式会社 Organic silicon compound and silane coupling agent containing same

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH08507293A (en) * 1993-01-28 1996-08-06 バイオコンパテイブルズ・リミテツド New material
JP2005187456A (en) * 2003-12-02 2005-07-14 Shiseido Co Ltd Phosphorylcholine group-containing compound and surface-modifying agent consisting of the same
CN101195638A (en) * 2007-12-24 2008-06-11 常州市绿空能源设备有限公司 Trinuclear phosphamide ester compounds containing tegafur and aza-silicon
JP2011236345A (en) * 2010-05-11 2011-11-24 Shiseido Co Ltd Surface modification method and surface-modified material
WO2013038901A1 (en) * 2011-09-16 2013-03-21 日産化学工業株式会社 Organic silicon compound and silane coupling agent containing same

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
YONG-ZHI OIU ET AL.: "A NOVEL ZWITTERIONIC SILANE COUPLING AGENT FOR NONTHROMBOGENIC BIOMATERIALS", CHINESE JOURNAL OF POLYMER SCIENCE, vol. 23, no. 6, 2005, pages 611 - 617 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2957563A1 (en) * 2014-06-19 2015-12-23 Pentracor GmbH Separation material comprising phosphoryl choline derivatives
WO2015193504A1 (en) * 2014-06-19 2015-12-23 Pentracor Gmbh Separation material comprising phosphoryl choline derivatives
CN106459108A (en) * 2014-06-19 2017-02-22 彭特科尔有限公司 Separation material comprising phosphoryl choline derivatives
RU2666357C2 (en) * 2014-06-19 2018-09-07 Пентракор Гмбх Separation material comprising phosphoryl choline derivatives
CN106459108B (en) * 2014-06-19 2019-10-15 彭特科尔有限公司 Separation material comprising Phosphorylcholine derivative

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