Compound, polymer, coating agent and tool

Abstract

To provide a polymer having biocompatibility.SOLUTION: There are provided: a 5- to 7-membered compound having an ethoxycarbonylalkyl group represented by 2-(2-ethoxycarbonylethyl)-2-oxazoline at the 2-position and having only one nitrogen atom and one oxygen atom as heteroatoms at the 1- and 3-positions; and a polymer thereof.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to organic chemistry and relates to compounds, polymers, coatings, and devices. [Background technology]

[0002] Development is underway for minimally invasive artificial organs for medical treatment and home diagnostic devices that are highly safe and reduce the burden on patients. Many medical devices contain polymeric materials. When blood comes into contact with a polymeric material that is not hemocompatible, it can activate biological defense mechanisms such as the contact phase, coagulation system, leukocyte system, platelet system, and complement system. Therefore, hemocompatibility is required for polymeric materials that come into direct contact with blood. Furthermore, biocompatibility is required for polymeric materials that come into contact with living organisms, not just blood (see, for example, Patent Documents 1 and 2). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] International Publication No. 2023 / 054500 [Patent Document 2] International Publication No. 2023 / 162926 Summary of the Invention [Problem to be solved by the invention]

[0004] An object of the present invention is to provide a biocompatible polymer. [Means for solving the problem]

[0005] [1] The compound according to an embodiment of the present invention is represented by the following general formula (1). [ka] (In the above general formula (1), m is a number of 1 or more and 3 or less, and L is a substituted or unsubstituted alkylene chain having 2 or more and 6 or less carbon atoms.)

[0006] [2] A polymer according to an embodiment of the present invention is a polymer of the compound described in [1].

[0007] [3] The polymer described in [2] above may have a unit represented by the following general formula (2): [ka] (In the above general formula (2), m is a number of 1 or more and 3 or less, and L is a substituted or unsubstituted alkylene chain having 2 or more and 6 or less carbon atoms.)

[0008] [4] The coating agent according to an embodiment of the present invention contains the polymer according to [2] or [3].

[0009] [5] The hydrophilic coating agent according to an embodiment of the present invention contains the polymer according to [2] or [3].

[0010] [6] A biocompatible coating agent according to an embodiment of the present invention contains the polymer according to [2] or [3].

[0011] [7] A medical coating agent according to an embodiment of the present invention contains the polymer according to [2] or [3].

[0012] [8] The implant coating agent according to an embodiment of the present invention contains the polymer according to [2] or [3].

[0013] [9] An antifouling coating agent according to an embodiment of the present invention contains the polymer according to [2] or [3].

[0014]

[10] In any one of the coating agents [4] to [9] above, the solvent may be ethanol.

[0015]

[11] An agent for improving the adhesion of cancer cells according to an embodiment of the present invention comprises the polymer described in [2] or [3].

[0016]

[12] A drug carrier according to an embodiment of the present invention comprises the polymer according to [2] or [3].

[0017]

[13] The coating agent according to an embodiment of the present invention contains the compound described in [1].

[0018]

[14] The hydrophilic coating agent according to an embodiment of the present invention comprises the compound described in [1].

[0019]

[15] A biocompatible coating agent according to an embodiment of the present invention includes the compound described in [1].

[0020]

[16] A medical coating agent according to an embodiment of the present invention comprises the compound described in [1].

[0021]

[17] The implant coating agent according to an embodiment of the present invention contains the compound described in [1].

[0022]

[18] The antifouling coating agent according to an embodiment of the present invention comprises the compound described in [1].

[0023]

[19] Any of the coating agents

[13] to

[18] above may further contain a polymerization initiator.

[0024]

[20] In any one of the coating agents

[13] to

[19] above, the solvent may be ethanol.

[0025]

[21] A medical device according to an embodiment of the present invention comprises a substrate and a layer containing the polymer according to [2] or [3] disposed on the substrate.

[0026]

[22] A biosensor according to an embodiment of the present invention includes a substrate and a layer containing the polymer according to [2] or [3] disposed on the substrate.

[0027]

[23] A culture substrate according to an embodiment of the present invention comprises a substrate and a layer containing the polymer according to [2] or [3] disposed on the substrate.

[0028]

[24] A method for producing a compound represented by the general formula (1) according to an embodiment of the present invention comprises ring-closing ethyl X—(Y-haloalkyl)amino-X-oxoalkanoate (wherein X=L+2, Y=m+1, L is a number between 2 and 6, and m is a number between 1 and 3).

[0029]

[25] In the method for producing the compound described in the above

[24] , the ring-closing reaction may be carried out in the presence of a base.

[0030]

[26] A method for producing a polymer according to an embodiment of the present invention comprises subjecting a compound represented by the above general formula (1) to a ring-opening isomerization polymerization reaction.

[0031]

[27] In the method for producing a polymer according to the above

[26] , the ring-opening isomerization polymerization reaction may be carried out in the presence of a polymerization initiator. [Effects of the Invention]

[0032] According to the present invention, a biocompatible polymer can be provided. [Brief explanation of the drawings]

[0033] [Figure 1] 1 is a graph showing the 1H-NMR spectrum of the monomer in deuterated chloroform (CDCl3) according to Example 2. [Figure 2] 1 is a graph showing the 1H-NMR spectrum of the polymer according to Example 3. [Figure 3] 1 is a graph showing a gel permeation chromatography trace of the polymer according to Example 3. [Figure 4] 10 is a graph showing the ratio of the intermediate water amount and the ratio of the non-freezing water amount in the polymer according to Example 5. [Figure 5] 10 is a graph showing the water contact angle on the substrate according to Example 7. [Figure 6] 10 is a photograph of a droplet on a substrate according to Example 7. [Figure 7] 10 is a graph showing the amount of BSA adsorbed to the substrate after 10 minutes of incubation according to Example 8. [Figure 8] 10 is a graph showing the amount of BSA adsorbed to the substrate after 1 hour of incubation according to Example 8. [Figure 9] 10 is a graph showing the amount of fibrinogen adsorbed to the substrate after 10 minutes of incubation according to Example 8. [Figure 10] 10 is a graph showing the amount of fibrinogen adsorbed to the substrate after 1 hour of incubation according to Example 8. [Figure 11] 10 is a graph showing the amount of fibronectin adsorbed to the substrate after 10 minutes of incubation according to Example 8. [Figure 12] 10 is a graph showing the amount of fibronectin adsorbed to the substrate after 1 hour of incubation according to Example 8. [Figure 13] 10 is a graph showing the amount of platelet-poor plasma protein adsorbed to a substrate after 10 minutes of incubation according to Example 8. [Figure 14] 10 is a graph showing the amount of platelet-poor plasma protein adsorbed to a substrate after 1 hour of incubation according to Example 8. [Figure 15] 10 is a graph showing the amount of γ chain presented by denaturation of fibrinogen adsorbed to a substrate in Example 9. [Figure 16] 10 is a graph showing the amount of RGD sequence presented by fibronectin adsorbed to a substrate in Example 9. [Figure 17] 10 is a graph showing the number of platelets adsorbed onto the substrate according to Example 10. [Figure 18] 10 is a photograph of platelets adsorbed on a substrate according to Example 10. [Figure 19] 10 is a graph showing the water contact angle on the substrate after immersion in PBS for a predetermined time according to Example 11. [Figure 20]12 is a graph showing the number of platelets adsorbed onto a substrate after immersion in PBS for a predetermined period of time according to Example 12. [Figure 21] 13 is a graph showing the number of cancer cells adhered to the substrate in Example 13. DETAILED DESCRIPTION OF THE INVENTION

[0034] The following describes embodiments of the present invention. However, it should not be understood that the following embodiments limit the present invention. From this disclosure, various alternative embodiments, examples, and operating techniques will become apparent to those skilled in the art. It should be understood that the present invention encompasses various embodiments not described herein.

[0035] The polymer according to the embodiment of the present invention is a polymer obtained by polymerizing a compound represented by the following general formula (1), and has a unit represented by the following general formula (2). [ka] In the above general formula (1), m is a number of 1 or more and 3 or less, preferably 1 or 2, and more preferably 1. L is a substituted or unsubstituted alkylene chain having 2 or more and 6 or less carbon atoms. [ka] In the above general formula (2), m is a number of 1 to 3, preferably 1 or 2, and more preferably 1. L is a substituted or unsubstituted alkylene chain having 2 to 6 carbon atoms. The number of carbon atoms in the alkylene chain is preferably 2 to 4, and more preferably 2. When the alkylene chain is substituted, examples of the substituent include halogen and an alkyl group having 1 to 4 carbon atoms.

[0036] When m is 1 and L is 2 in the above general formula (1), the compound is 2-(2-ethoxycarbonylethyl)-2-oxazoline, which is represented by the following general formula (3). The compound represented by the following general formula (3) is also called ethyl 3-(4,5-dihydrooxazol-2-yl)propanoate. When m is 2 and L is 2 in the above general formula (1), the compound is ethyl 3-(5,6-dihydro-4H-1,3-oxazine) or ethyl 3-(5,6-dihydro-4H-1,3-oxazin-2-yl)propanoate. When m is 3 and L is 2 in the above general formula (1), the compound is 2-(2-ethoxycarbonylethyl)-2-(4,5,6,7-tetrahydro-1,3-oxazepin) or ethyl 3-(4,5,6,7-tetrahydro-1,3-oxazepin-2-yl)propanoate. [ka]

[0037] When m is 1 and L is an ethylene chain in the above general formula (2), the polymer is poly[2-(2-ethoxycarbonylethyl)-2-oxazoline], which has units represented by the following general formula (4). The polymer according to an embodiment of the present invention is also called poly[ethyl 3-(4,5-dihydrooxazol-2-yl)propanoate]. The polymer according to an embodiment of the present invention is also referred to as PC2EtstOx. [ka]

[0038] The number average molecular weight (Mn) of the polymer according to an embodiment of the present invention is, for example, 10,000 to 50,000, 10,000 to 40,000, 10,000 to 30,000, 10,000 to 20,000, 10,000 to 17,000, 11,000 to 16,000, or 12,000 to 15,000. The molecular weight dispersity (PDI=Mw / Mn) of the polymer according to an embodiment of the present invention is, for example, 1.00 to 4.00, 1.01 to 3.00, 1.01 to 2.00, or 1.01 to 1.60.

[0039] The compound represented by the above general formula (1) can be obtained by ring-closing ethyl X-(Y-haloalkyl)amino-X-oxoalkanoate (where X=L+2, Y=m+1, L is a number from 2 to 6, and m is a number from 1 to 3), as shown in the following reaction scheme. The ethyl X-(Y-haloalkyl)amino-X-oxoalkanoate is preferably ethyl 4-(2-haloalkyl)amino-4-oxobutanoate. The halogen in the haloalkyl is fluorine, chlorine, bromine, iodine, or astatine. The haloalkyl is preferably chloroalkyl. [ka]

[0040] 2-(2-Ethoxycarbonylethyl)-2-oxazoline can be obtained by ring-closing ethyl 4-(2-haloethyl)amino-4-oxobutanoate, as shown in the following reaction formula. The halogen in haloethyl is fluorine, chlorine, bromine, iodine, or astatine. The haloethyl is preferably chloroethyl. The ring-closing reaction may be carried out in the presence of a base. Examples of the base include alkali metal salts such as sodium carbonate (NaCO), sodium hydroxide (NaOH), potassium hydroxide (KOH), and potassium carbonate (KCO). 18-Crown-6 may also be added to the ring-closing reaction. [ka]

[0041] The polymer according to the embodiment of the present invention can be obtained by ring-opening polymerization of the compound represented by the above general formula (1). The polymer according to the embodiment of the present invention can be preferably obtained by ring-opening polymerization of 2-(2-ethoxycarbonylethyl)-2-oxazoline, as shown in the following reaction formula: [ka]

[0042] The ring-opening isomerization polymerization reaction may be carried out in the presence of a polymerization initiator. Examples of the polymerization initiator include sulfonic acid derivatives, alkyl halides, benzyl halides, and allyl halides. Examples of sulfonic acid derivatives include methyl tosylate.

[0043] Polymers according to embodiments of the present invention can be dissolved in a variety of solvents, including methanol, ethanol, toluene, benzene, chloroform, dichloromethane, acetone, acetonitrile, tetrahydrofuran (THF), 2-propanol, dimethyl sulfoxide (DMSO), dimethylformamide (DMF), 1,4-dioxane, ethyl acetate, dimethylacetamide (DMAC), and N-methyl-2-pyrrolidone (NMP).

[0044] The coating agent according to an embodiment of the present invention contains the above-described polymer. The coating agent containing the polymer according to an embodiment of the present invention can be produced by dissolving the above-described polymer in a solvent. By coating a substrate with the coating agent containing the polymer according to an embodiment of the present invention, it is possible to impart hydrophilicity and biocompatibility to the substrate.

[0045] A coating agent according to another embodiment of the present invention comprises the compound described above. The coating agent comprising the compound described above may further comprise a polymerization initiator. The coating agent comprising the compound according to an embodiment of the present invention can be produced by dissolving the compound described above in a solvent. Hydrophilicity and biocompatibility can be imparted to a substrate by polymerizing the compound while coating the substrate with a coating agent comprising the compound according to an embodiment of the present invention, or by coating the substrate with a coating agent comprising the compound according to an embodiment of the present invention and then polymerizing the compound.

[0046] By coating a substrate with the coating agent according to an embodiment of the present invention, it is possible to inhibit the adsorption of proteins such as albumin, fibrinogen, and fibronectin to the substrate. Furthermore, by coating a substrate with the coating agent according to an embodiment of the present invention, it is possible to inhibit the adsorption of platelets to the substrate. By coating a substrate with the coating agent according to an embodiment of the present invention, it is possible to impart antithrombotic properties to the substrate. By coating a substrate with the coating agent according to an embodiment of the present invention, it is possible to promote the adhesion of cancer cells to the substrate.

[0047] By coating an instrument with a coating agent according to an embodiment of the present invention, it is possible to impart biocompatibility, hydrophilicity, antithrombogenicity, and antifouling properties to the instrument. The instrument may be included in a device. Examples of instruments and devices include dental instruments such as implants, stents, catheters, blood bags, transfusion instruments, surgical instruments, blood circulation devices, blood purification devices, plasma separation devices, artificial blood vessels, and artificial organs such as heart-lung machines and artificial kidneys. The instrument may also be a biosensor.

[0048] The polymer according to the embodiment of the present invention can be used as a drug carrier in a drug delivery system. For example, since the polymer according to the embodiment of the present invention has affinity for cancer cells, it can be used as a carrier of a cancer therapeutic drug. [Example]

[0049] Example 1: Synthesis of Monomer Precursor Ethyl succinyl chloride (100 g, 0.607 mol) and 2-chloroethylamine hydrochloride (71.16 g, 0.607 mol) were suspended in dry dichloromethane (1,000 mL) under an argon atmosphere. Triethylamine (EtN) (1.335 mol, 187.62 mL) was added dropwise over 2 hours while the reaction mixture was continuously cooled to 0 °C. The reaction mixture was allowed to warm to room temperature overnight. After the reaction, 160 mL of water was added, and the organic phase was separated from the aqueous phase. The organic phase was extracted with 160 mL of water (twice) and 160 mL of saturated brine (once). The organic phase was then dried over MgSO and concentrated in vacuo to give the monomer precursor, ethyl 4-(2-chloroethyl)amino-4-oxobutanoate. [ka] Scheme 1: Synthetic route to monomer precursors

[0050] Example 2: Synthesis of Monomer Sodium carbonate (Na2CO3) was dried at 180 °C for 24 h. Next, Na2CO3 (1 eq.) was added to the monomer precursor in a round-bottom flask. The ring-closure reaction of the monomer precursor was carried out using a rotary evaporator at 50 mbar and 40 °C. The conversion of the monomer precursor to the monomer, 2-(2-ethoxycarbonylethyl)-2-oxazoline, was carried out as follows: 1 The reaction mixture was monitored by H-NMR. 2-(2-ethoxycarbonylethyl)-2-oxazoline is also referred to as C2EtstOx below. After 4 days, 90% conversion was confirmed. Dichloromethane was then added to the reaction mixture, and Na2CO3 was removed by filtration. The solvent was evaporated to obtain the crude monomer. The monomer was further distilled under reduced pressure three times using barium oxide (BaO) and ninhydrin. The monomer in deuterated chloroform (CDCl3) was 1 The 1 H NMR spectrum is shown in Figure 1. [ka] Scheme 2: Synthetic route to the monomer

[0051] Example 3: Polymer synthesis A reaction mixture containing the monomer and methyl tosylate as an initiator was prepared in acetonitrile to a monomer concentration of 3 mol / L. The polymerization was carried out at 45°C for 4 days. The conversion of the monomer to polymer was 1 It was followed by H-NMR. 1 The H NMR spectrum is shown in Figure 2. The conversion rate from monomer to polymer reached 84%. The polymerization reaction was terminated by adding dry sodium azide (10 eq. per methyl tosylate). The mixture was then stirred at room temperature for 2 days. The sodium azide was removed by syringe filtration to isolate the polymer. The filtrate was then precipitated in cold diethyl ether and dried under vacuum to obtain the polymer poly[2-(2-ethoxycarbonylethyl)-2-oxazoline] (PC2EtstOx). [ka] Scheme 3: Polymer synthesis route

[0052] The molecular weight of PC2EtstOx was measured by gel permeation chromatography (GPC) at 40 °C using an HPLC system (Prominence, Shimadzu) equipped with four columns (TSKgel, Tosoh) and a refractive index detector. The eluent was dimethylformamide containing 10 mmol / L LiBr at a flow rate of 1.0 mL / min. The gel permeation chromatography trace is shown in Figure 3. The number-average molecular weight (Mn) of PC2EtstOx was 14,300. The weight-average molecular weight (Mw) of PC2EtstOx was 15,100. The polydispersity index (PDI), given by Mw / Mn, was 1.06.

[0053] Example 4: Evaluation of the solubility of PC2EtstOx The PC2EtstOx obtained in Example 3 was added to water, methanol, ethanol, toluene, benzene, chloroform, dichloromethane, acetone, acetonitrile, tetrahydrofuran (THF), 2-propanol, dimethyl sulfoxide (DMSO), dimethylformamide (DMF), 1,4-dioxane, ethyl acetate, dimethylacetamide (DMAC), N-methyl-2-pyrrolidone (NMP), diethyl ether, and hexane at a final concentration of 10 mg / mL. These mixtures were allowed to stand at room temperature for 4 hours, and the solubility was measured. As shown in Tables 1 to 4, PC2EtstOx was soluble in methanol, ethanol, toluene, benzene, chloroform, dichloromethane, acetone, acetonitrile, tetrahydrofuran (THF), 2-propanol, dimethyl sulfoxide (DMSO), dimethylformamide (DMF), 1,4-dioxane, ethyl acetate, dimethylacetamide (DMAC), and N-methyl-2-pyrrolidone (NMP). PC2EtstOx was insoluble in water, diethyl ether, and hexane. Generally, poly(2-methoxyethyl acrylate) (PMEA) is known to be insoluble in ethanol, and the PC2EtstOx of the present example was confirmed to be soluble in ethanol. [Table 1] [Table 2] [Table 3] [Table 4]

[0054] Example 5: Evaluation of hydration water of PC2EtstOx by DSC It is known that water saturated in polymers can be classified into free water, non-freezing water, and intermediate water. Pure water exhibits a melting peak at 0°C. Although free water is adsorbed to the polymer, it has properties similar to bulk water and has a weak interaction with the polymer, melting at around 0°C. Non-freezing water has a strong interaction with the polymer and does not freeze even at -100°C. Intermediate water is neither free water nor non-freezing water, but has an intermediate interaction with the polymer, forming crystals at temperatures below 0°C and melting at temperatures below 0°C.

[0055] The amount of water of hydration contained in the PC2EtstOx obtained in Example 3 was quantified using a differential scanning calorimeter (X-DSC7000, Seiko). The PC2EtstOx obtained in Example 3 was immersed in a large excess of pure water (10 g of pure water per 30 mg of PC2EtstOx) and allowed to stand at room temperature (25°C) for 3 days to allow it to absorb water. The hydrated PC2EtstOx was removed from the water using tweezers, and water adhering to the hydrated PC2EtstOx was removed using a medical paper. Then, 3 to 5 mg of PC2EtstOx was placed on an aluminum pan and weighed. The weighed PC2EtstOx was cooled from 30°C to −100°C using a sealed differential scanning calorimeter, held at −100°C for 5 minutes, and then heated to 50°C at a rate of 5°C / min. Based on the observed exothermic transition during the low-temperature crystallization of water, the water contained in the hydrated PC2EtstOx was classified into intermediate water, free water, and non-freezable water according to the following method, and the intermediate water and non-freezable water were quantified.

[0056] Water whose exothermic peak temperature was observed at -35°C or below during the temperature-lowering (cooling) process in measurements using a differential scanning calorimeter, and water whose exothermic peak temperature was observed during the temperature-raising (heating) process, were both considered intermediate water. Water whose exothermic peak temperature was observed at a temperature above -35°C during the temperature-lowering process was considered free water. Water that did not form low-temperature crystals even at -100°C, the lower limit of the temperature range for measurements using a differential scanning calorimeter, was considered nonfreezing water.

[0057] After measurement using a differential scanning calorimeter, holes were made in the aluminum pan, and the PC2EtstOx was vacuum-dried at 110°C. The mass (unit: g) of the dried PC2EtstOx was measured. The change in mass of PC2EtstOx before and after drying was taken as the amount of hydration water (unit: g) contained in PC2EtstOx. The intermediate water, free water, and unfrozen water were quantified according to the following equations (1) to (3). Intermediate water volume = Total enthalpy change of intermediate water ÷ Latent heat of fusion of water (1) Amount of free water = Total enthalpy change of free water ÷ Latent heat of fusion of water (2) Unfrozen water volume = hydration water volume of PC2EtstOx - intermediate water volume - free water volume (3) In equations (1) to (3), the units for the intermediate water volume, free water volume, and unfrozen water volume are g, the unit for the enthalpy phase change is J, and the latent heat of fusion of water is 334 J / g. The total enthalpy change of the intermediate water was calculated based on the sum of the peak area of ​​the exothermic peak observed at temperatures below -35°C during the cooling process and the peak area of ​​the exothermic peak observed during the heating process.

[0058] As a control, the amount of intermediate water and the amount of nonfreezing water in poly(2-methoxyethyl acrylate) (PMEA) were measured using the same method. As shown in Figure 4, the amount of intermediate water per unit polymer weight (1 g) of PC2EtstOx obtained in Example 3 was significantly higher than the amount of intermediate water per unit polymer weight (1 g) of PMEA. Furthermore, the amount of nonfreezing water per unit polymer weight (1 g) of PC2EtstOx obtained in Example 3 was significantly higher than the amount of nonfreezing water per unit polymer weight (1 g) of PMEA.

[0059] Example 6: Preparation of PC2EtstOx-coated substrate Substrates made of polystyrene (PS), glass, polycarbonate (PC), polyethylene (PE), polymethyl methacrylate (PMMA), polypropylene (PP), and polyethylene terephthalate (PET) were prepared. The PC2EtstOx solution of Example 6 was prepared by dissolving the PC2EtstOx obtained in Example 3 in ethanol at a concentration of 0.2% w / v. Using a spin coater (Mikasa, MS-A100), the PC2EtstOx solution of Example 6 was spin-coated onto each substrate twice. The spin-coating conditions for each cycle were: 500 rpm for 5 seconds, 2000 rpm for 10 seconds, ramping up to 4000 rpm for 5 seconds, 4000 rpm for 5 seconds, and ramping down for 4 seconds. The substrates were then dried in an incubator (AS ONE, WI-50), forming coating layers on the substrates.

[0060] As a control, PMEA was dissolved in methanol at a concentration of 0.2% w / v to obtain a PMEA solution, which was then spin-coated onto the substrate using the same method as above.

[0061] Example 7: Measurement of water contact angle A 2 μL water droplet was placed on each of a substrate coated with the PC2EtstOx solution of Example 6, a substrate coated with the PMEA solution, and an uncoated substrate, and the water contact angle was measured by the sessile drop method. As shown in FIG. 5, when the PMEA solution was coated, the water contact angle decreased compared to before coating on all of the substrates, improving hydrophilicity. Furthermore, when the PC2EtstOx solution of Example 6 was coated, the water contact angle decreased compared to before coating on all of the substrates, improving hydrophilicity.

[0062] On polystyrene (PS) substrates, glass substrates, polycarbonate (PC) substrates, polymethyl methacrylate (PMMA) substrates, polypropylene (PP) substrates, and polyethylene terephthalate (PET) substrates, PC2EtstOx obtained in Example 3 improved hydrophilicity more than PMEA. On polyethylene (PE) substrates, PMEA improved hydrophilicity more than PC2EtstOx obtained in Example 3. Photographs of droplets on uncoated polyethylene terephthalate (PET) substrates, PMEA-coated PET substrates, and PC2EtstOx-coated PET substrates obtained in Example 3 are shown in Figure 6.

[0063] Example 8: Evaluation of protein adsorption A 96-well cell culture polystyrene (TCPS) plate (IWAKI) was prepared. 15 μL of the PC2EtstOx solution according to Example 6 was added to each well. As a control, 15 μL of the PMEA solution according to Example 6 was added to each well of the plate. The plate was gently air-dried at room temperature for 3 days.

[0064] Proteins were prepared using bovine serum albumin (BSA), fibrinogen, and fibronectin, and 1 mg / mL protein solutions were prepared. Platelet-poor plasma (PPP) was also prepared. 50 μL of either the protein solution or platelet-poor plasma was added to each well of the plate and incubated at 37°C for 10 minutes or 1 hour. After incubation, the wells were washed five times with PBS and then treated with a solution of 5% sodium dodecyl sulfate (SDS) and 0.1 N NaOH for 60 minutes at 37°C.

[0065] The amount of protein adsorbed to the wells was then measured using a total protein quantification kit (Micro BCA Protein Assay Kit, Thermo Scientific). As shown in Figures 7 and 8, less BSA was adsorbed to the wells coated with PC2EtstOx than to the wells coated with PMEA. As shown in Figures 9 and 10, less fibrinogen was adsorbed to the wells coated with PC2EtstOx than to the wells coated with PMEA. As shown in Figures 11 and 12, less fibronectin was adsorbed to the wells coated with PC2EtstOx than to the wells coated with PMEA. As shown in Figures 13 and 14, less proteins contained in platelet-poor plasma were adsorbed to the wells coated with PC2EtstOx than to the wells coated with PMEA.

[0066] Example 9: ELISA Assay The conformational changes of fibrinogen and fibronectin adsorbed to the plate were evaluated by ELISA assay. Wells coated with PC2EtstOx and PMEA were prepared using the same method as in Example 8. Each well of the plate was loaded with 50 μL of either platelet-poor plasma or 10 μg / mL fibronectin and incubated at 37°C for 1 hour. Note that platelet-poor plasma is rich in fibrinogen. After incubation, the wells were washed five times with PBS and then treated with blocking solution (Blocking One, Nacalai Tesque) at room temperature for 1 hour.

[0067] The wells were incubated with mouse anti-human fibronectin RGD sequence antibody (M002; Takara Bio) or mouse anti-human fibrinogen γ-chain antibody (SC-133157; Santa Cruz Biotechnology) for 90 minutes at room temperature. The wells were then incubated with a secondary antibody (HRP-conjugated goat anti-mouse IgG, STAR207P; Bio-Rad) for 2 hours at 37°C. The wells were then rinsed five times with PBS and incubated with ABTS solution (1 mg / mL) for 10 minutes at 37°C. The absorbance at 405 nm was then measured using a microplate reader (Infinite 200 PRO M Plex, Tecan).

[0068] As shown in Figure 15, the amount of fibrinogen γ-chain displayed due to conformational changes was low in both PC2EtstOx-coated wells and PMEA-coated wells. Furthermore, as shown in Figure 16, the amount of fibronectin RGD sequence displayed was higher in both PC2EtstOx-coated wells and PMEA-coated wells than in untreated wells. The amount of fibronectin RGD sequence displayed in PC2EtstOx-coated wells was lower than in PMEA-coated wells.

[0069] Example 10: Human platelet adsorption test Platelet-rich plasma (PRP) and platelet-poor plasma (PPP) were obtained from human whole blood by two-step centrifugation. PRP and PPP were then mixed to obtain 4 × 10 7 pieces / cm 2A plasma solution containing 100 μL of platelets was prepared. Substrates made of PMMA, PP, PC, and PET were also prepared. Each substrate was coated with PC2EtstOx or PMEA. 200 μL of plasma solution was applied to the surface of a PC2EtstOx or PMEA-coated substrate (8 mm × 8 mm square) and incubated at 37°C for 1 hour. The substrate was then rinsed with PBS and then immersed in a 1% glutaraldehyde solution in PBS for 120 minutes at 37°C. The substrate was then rinsed with PBS and Milli-Q water. The platelet adsorption profile on the substrate was evaluated using scanning electron microscopy (SEM).

[0070] As shown in Figure 17, platelet adsorption was significantly less on the substrates coated with PC2EtstOx or PMEA than on the uncoated substrates. The degree of platelet activation was also classified into spherical platelets (type I), round platelets with a small amount of pseudopodia (type II), and partially activated platelets (type III). Platelets can be classified into three types: flattened platelets with numerous pseudopodia (type III, activated platelets that are prone to thrombus formation), and activated platelets with many pseudopodia (type III, activated platelets that are prone to thrombus formation). On the PC2EtstOx- or PMEA-coated substrates, the degree of activity of most of the slightly adsorbed platelets was type I. Figure 18 shows SEM images of platelets on an untreated PET substrate, a PMEA-coated PET substrate, and a PC2EtstOx-coated PET substrate.

[0071] (Example 11: Measurement of water contact angle after PBS immersion) Substrates coated with PC2EtstOx and PMEA were prepared. The substrates were immersed in 37°C PBS for 1 hour, 6 hours, or 24 hours. The substrates were then dried in an incubator (AS ONE, WI-50), and the water contact angle of each substrate was measured using the same method as in Example 7. As shown in Figure 19, the contact angle of the PMEA-coated substrate increased after immersion in PBS, resulting in a decrease in hydrophilicity. In contrast, the contact angle of the PC2EtstOx-coated substrate increased only slightly after immersion in PBS, maintaining its hydrophilicity. This demonstrates that PC2EtstOx has high durability.

[0072] Example 12: Adsorption test of human platelets after PBS impregnation Substrates coated with PC2EtstOx and PMEA were prepared. The substrates were immersed in PBS at 37°C for 1 hour or 6 hours. The substrates were then dried in an incubator (AS ONE, WI-50), and the number of platelets adsorbed to each substrate was evaluated using the same method as in Example 10. As shown in Figure 20, the number of platelets adsorbed to the PMEA-coated substrate increased significantly after 6 hours of immersion in PBS. In contrast, the number of platelets adsorbed to the PC2EtstOx-coated substrate increased only slightly even after 6 hours of immersion in PBS.

[0073] Example 13: Cancer cell adhesion test A 24-well tissue culture polystyrene (TCPS) plate was coated with 40 μL of 0.5% w / v poly(n-butyl methacrylate-co-2-methacryloyloxyethyl phosphorylcholine) solution and allowed to dry overnight. Each well contained 14 mm diameter uncoated PET substrates, PC2EtstOx-coated PET substrates, and PMEA-coated PET substrates. The wells were then exposed to UV light for 1 hour for sterilization. The wells were then thoroughly rinsed with PBS, followed by addition of cell culture medium and incubation at 37°C for an additional 1 hour.

[0074] HepG2 cells, MDA-MB-231 cells, and MCF-7 cells were prepared as cancer cells. Each cancer cell was placed on the substrate in a well at a density of 1 × 10 4 cells / cm 2 Cells were seeded at a concentration of 100 μL each and allowed to adhere for 1 hour. HepG2 cells were cultured in DMEM / F12 containing 10% FBS, while MDA-MB-231 and MCF-7 cells were cultured in DMEM containing 10% FBS. The culture medium was replaced with 1000 μL of fresh medium, and 100 μL of a viable cell count kit (CCK-8, Dojindo) was added to each well. After incubation at 37°C for 2 hours, the absorbance at 405 nm in each well was measured using a microplate reader (Infinite 200 PRO M Plex, Tecan), and cell numbers were calculated using the standard curve method.

[0075] As a result, as shown in Figure 21, it was confirmed that a greater number of cancer cells adhered to the PET substrate coated with PC2EtstOx than to the PET substrate coated with PMEA.

Claims

1. A compound represented by the following general formula (1): 【Chemistry 1】 (In the above general formula (1), m is a number of 1 or more and 3 or less, and L is a substituted or unsubstituted alkylene chain having 2 or more and 6 or less carbon atoms.)

2. A polymer of the compound of claim 1.

3. The polymer according to claim 2, having a unit represented by the following general formula (2): 【Chemistry 2】 (In the above general formula (2), m is a number of 1 or more and 3 or less, and L is a substituted or unsubstituted alkylene chain having 2 or more and 6 or less carbon atoms.)

4. A coating agent comprising the polymer of claim 2.

5. An agent for improving the adhesion of cancer cells, comprising the polymer of claim 2.

6. A drug carrier comprising the polymer of claim 2.

7. A coating agent comprising the compound of claim 1.

8. The coating agent according to claim 12, further comprising a polymerization initiator.

9. A substrate; a layer comprising the polymer of claim 2 disposed on the substrate; A medical device comprising:

10. A substrate; a layer comprising the polymer of claim 2 disposed on the substrate; A biosensor comprising:

11. A substrate; a layer comprising the polymer of claim 2 disposed on the substrate; A culture substrate comprising:

12. cyclization of ethyl X-(Y-haloalkyl)amino-X-oxoalkanoate, where X=L+2, Y=m+1, L is a number between 2 and 6, and m is a number between 1 and 3; A method for producing a compound represented by the following general formula (1): 【Transformation 3】 (In the above general formula (1), m is a number of 1 or more and 3 or less, and L is a substituted or unsubstituted alkylene chain having 2 or more and 6 or less carbon atoms.)

13. The method for producing the compound according to claim 12, wherein the ring-closure reaction is carried out in the presence of a base.

14. A method for producing a polymer, comprising subjecting a compound represented by the following general formula (1) to a ring-opening isomerization polymerization reaction: 【Chemistry 4】 (In the above general formula (1), m is a number of 1 or more and 3 or less, and L is a substituted or unsubstituted alkylene chain having 2 or more and 6 or less carbon atoms.)

15. The method for producing a polymer according to claim 14 , wherein the ring-opening isomerization polymerization reaction is carried out in the presence of a polymerization initiator.

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