Composition for adhering cancer cells, cancer cell collection filter, and method for detecting cancer cells

Abstract

[Problem] To provide a polymer material that is biocompatible and, at the same time, has cancer cell adhesion properties. [Solution] A composition for cancer cell adhesion according to the present invention contains a biocompatible copolymer, said copolymer comprising at least one kind of repeating unit (A) represented by formula (1) (wherein R1 represents a methyl group or an ethyl group) and at least one kind of repeating unit (B) represented by formula (2) (wherein R2 represents an aliphatic hydrocarbon group).

Description

[Technical field]

[0001] The present invention relates to a composition for cancer cell adhesion. The present invention also relates to a cancer cell collection filter having the composition for cancer cell adhesion. Furthermore, the present invention also relates to a method for detecting cancer cells using the cancer cell collection filter. [Background technology]

[0002] Conventionally, medical materials using various polymeric materials have been studied. Since medical materials are used by contacting synthetic materials, which are foreign to the living body, with tissues and blood in the living body, they are required to be biocompatible. To date, for example, poly(2-methoxyethyl acrylate) (PMEA) has been developed as a biocompatible material (see Patent Document 1). This PMEA is known to exhibit biocompatibility due to the presence of so-called intermediate water (water in a state in which an exothermic peak due to low-temperature crystallization of water is stably observed around -40°C in differential scanning calorimetry during the temperature rise process from -100°C), which is considered to be water weakly bound to the polymer chain due to the interaction with the polymer chain.

[0003] In recent years, a copolymer of diethylene glycol monoethyl monovinyl ether (EOEOVE) as a hydrophilic vinyl ether and n-butyl vinyl ether as a hydrophobic vinyl ether has been proposed as a biocompatible material that has an oxyethylene chain structure in the side chain like PMEA and intermediate water (see Patent Document 2). However, this copolymer has been shown to have extremely low adhesiveness to cancer cells.

[0004] On the other hand, currently, there is a demand for polymeric materials that are biocompatible and have cancer cell adhesive properties in order to detect cancer cells. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] JP 2004-161954 A [Patent Document 2] International Publication No. 2017 / 150000 Summary of the Invention [Problem to be solved by the invention]

[0006] Therefore, an object of the present invention is to provide a polymeric material having both biocompatibility and cancer cell adhesiveness, and to provide a cancer cell collection filter and a cancer cell detection method using the polymeric material having cancer cell adhesiveness. [Means for solving the problem]

[0007] As a result of intensive research aimed at solving the above-mentioned problems, the present inventors have found that a copolymer containing a specific repeating unit (A) derived from a specific hydrophilic vinyl ether other than diethylene glycol monoethyl monovinyl ether (EOEOVE) and a repeating unit (B) derived from a specific hydrophobic vinyl ether has both biocompatibility and cancer cell adhesiveness, thereby completing the present invention.

[0008] That is, according to the present invention, the following inventions are provided. [1] At least one of the following formula (1): [ka] (In the formula, R 1 indicates a methyl or ethyl group) A repeating unit (A) represented by the formula: At least one of the following formula (2): [ka] (In the formula, R 2 indicates an aliphatic hydrocarbon group) and a repeating unit (B) represented by the following formula (1): [2] The cancer cell adhesion composition described in [1], wherein the biocompatible copolymer is a random copolymer of segment A consisting of repeating unit (A) and segment B consisting of repeating unit (B). [3] The cancer cell adhesion composition according to [1] or [2], wherein the composition ratio (molar ratio) of the repeating unit (A) to the repeating unit (B) in the biocompatible copolymer is 90:10 to 10:90. [4] R in the repeating unit (B) of the biocompatible copolymer 2 is a linear or branched alkyl or alkenyl group having 2 to 10 carbon atoms, or a monocyclic or polycyclic alkyl or alkenyl group having 3 to 20 carbon atoms. [5] R in the repeating unit (B) of the biocompatible copolymer 2 The composition for cancer cell adhesion according to any one of [1] to [3], wherein is a linear or branched alkyl group having 2 to 10 carbon atoms. [6] The composition for cancer cell adhesion according to any one of [1] to [5], wherein the ratio of the weight average molecular weight (Mw) to the number average molecular weight (Mn) of the biocompatible copolymer is 1.0 to 3.0. [7] The cancer cell adhesion composition according to any one of [1] to [6], wherein the number average molecular weight (Mn) of the biocompatible copolymer is 3,000 to 30,000. [8] A cancer cell collecting filter having the composition for cancer cell adhesion according to any one of [1] to [7] on at least a part of its surface. [9] A method for detecting cancer cells, comprising: A method for detecting cancer cells, comprising a step of filtering blood using the cancer cell collection filter described in [8].

[10] The method for detecting cancer cells described in [9], further comprising a step of culturing the cancer cells collected in the filtration step. Effect of the Invention

[0009] According to the present invention, it is possible to provide a polymeric material having both biocompatibility and cancer cell adhesiveness. Also, according to the present invention, it is possible to provide a cancer cell collection filter and a cancer cell detection method using the polymeric material having cancer cell adhesiveness. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] <Composition for cancer cell adhesion> The cancer cell adhesion composition according to the present invention comprises a biocompatible copolymer containing at least one type of repeating unit (A) and at least one type of repeating unit (B).

[0011] (Biocompatible copolymer) The biocompatible copolymer of the present invention comprises at least one copolymer represented by the following formula (1): [ka] (In the formula, R 1 indicates a methyl or ethyl group) A repeating unit (A) represented by the formula: At least one of the following formula (2): [ka] (In the formula, R 2 indicates an aliphatic hydrocarbon group) and a repeating unit (B) represented by the following formula (1):

[0012] The monomer that provides the repeating unit (A) of the formula (1) is a monomer represented by the following formula (3): [ka] (In the formula, R 1 is R in formula (1). 1 is equivalent to Examples of the hydrophilic vinyl ether include those represented by the following formula:

[0013] More specifically, hydrophilic vinyl ethers represented by formula (3) include 2-methoxyethyl vinyl ether (MOVE) and 2-ethoxyethyl vinyl ether (EOVE). Among these, MOVE is preferred because of its superior cancer cell adhesiveness.

[0014] The monomer that provides the repeating unit (B) is represented by the following formula (4): [ka] (In the formula, R 2 is R in Eq. (2). 2 is equivalent to In this case, hydrophobic vinyl ethers represented by the formula: 2 is an aliphatic hydrocarbon group, specifically, a linear or branched alkyl or alkenyl group, or a monocyclic or polycyclic alkyl or alkenyl group.

[0015] The above linear or branched alkyl or alkenyl group preferably has 2 to 10 carbon atoms, more preferably has 3 to 8 carbon atoms, and further preferably has 4 to 6 carbon atoms.

[0016] Specific examples of linear or branched alkyl or alkenyl groups include, for example, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a tert-butyl group, a 1-pentyl group, a 2-pentyl group, a 3-pentyl group, a 1-(2-methyl)-butyl group, a 2-(2-methyl)-butyl group, a 1-(3-methyl)-butyl group, a 2-(3-methyl)-butyl group, a (2,2-dimethyl)-propyl group, a 1-hexyl group, a 2-hexyl group, a 3-hexyl group, a 1-heptyl group, a 2-heptyl group, a 2-hex ... linear or branched alkyl groups such as butyl, 3-heptyl, 4-heptyl, 1-octyl, and 1-(2-ethyl)-hexyl groups; and linear or branched alkenyl groups such as vinyl, 1-propenyl, allyl, 2-butenyl, 3-butenyl, isopropenyl, isobutenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, and 5-hexenyl groups.

[0017] Furthermore, the monocyclic or polycyclic alkyl or alkenyl group preferably has 3 to 20 carbon atoms, more preferably has 4 to 15 carbon atoms, and further preferably has 5 to 12 carbon atoms.

[0018] Specific examples of the monocyclic or polycyclic alkyl or alkenyl group include monocyclic alkyl or alkenyl groups such as a cyclopentyl group, a cyclopentylmethyl group, a methylcyclopentyl group, a dimethylcyclopentyl group, a cyclohexyl group, a cyclohexylmethyl group, a methylcyclohexyl group, a dimethylcyclohexyl group, a cyclohexenyl group, a cycloheptyl group, a cyclooctyl group, a cyclononyl group, a cyclodecyl group, a cycloundecyl group, a cyclododecyl group, a cyclotridecyl group, a cyclotetradecyl group, a cyclopentadecyl group, a cyclooctadecyl group, and a cycloicosyl group; and polycyclic alkyl or alkenyl groups such as a bicyclohexyl group, a decahydronaphthyl group, a norbornyl group, a methylnorbornyl group, an isobornyl group, an adamantyl group, a tricyclodecanyl group, a tricyclodecenyl group, and a tetracyclododecyl group.

[0019] Among these aliphatic hydrocarbon groups, linear or branched alkyl groups are preferred. More specifically, as the hydrophobic vinyl ether represented by formula (4), n-butyl vinyl ether (NBVE) and 2-ethylhexyl vinyl ether (EHVE) are preferred. Among these, NBVE is preferred because it has better cancer cell adhesion.

[0020] The composition ratio (molar ratio) of the repeating unit (A) to the repeating unit (B) in the biocompatible copolymer of the present invention (hereinafter sometimes referred to as "copolymer") is preferably in the range of 90:10 to 10:90, more preferably in the range of 85:15 to 15:85, and particularly preferably in the range of 80:20 to 20:80.

[0021] The molecular weight of the copolymer in the present invention, as the number average molecular weight (Mn) determined from a standard polystyrene calibration curve by gel permeation chromatography (GPC), is preferably 1,000 to 100,000, more preferably 2,000 to 50,000, and even more preferably 3,000 to 30,000. If Mn is within the above range, the copolymer will have better adhesiveness to cancer cells.

[0022] The ratio of weight average molecular weight (Mw) to number average molecular weight (Mn) of the copolymer in the present invention (molecular weight distribution, Mw / Mn) is preferably 1.0 to 3.0, more preferably 1.0 to 2.0, even more preferably 1.0 to 1.5, and even more preferably 1.0 to 1.3. If the molecular weight distribution (Mw / Mn) is within the above range, the copolymer will have better adhesiveness to cancer cells.

[0023] The copolymer in the present invention may be either a random copolymer or a block copolymer, but from the viewpoint of adhesiveness to cancer cells, a random copolymer containing a segment A consisting of a repeating unit (A) and a segment B consisting of a repeating unit (B) is preferred. The composition distribution of the segments A and B in the random copolymer is not particularly limited, and examples thereof include a completely random copolymer having a statistically random composition and a tapered (gradient) random copolymer having a gradient in composition distribution. When the copolymer is a random copolymer, sufficient adhesiveness to cancer cells can be obtained regardless of the molecular weight.

[0024] The copolymer of the present invention can be prepared by polymerizing the hydrophilic vinyl ether represented by the above formula (3) and the hydrophobic vinyl ether (4) represented by the above formula (4) according to a conventional method. As the polymerization method, living cationic polymerization is particularly preferred in order to obtain a copolymer with a desired composition ratio and molecular weight with good reproducibility. In the living cationic polymerization method, the molecular weight of the copolymer is almost uniquely determined by the molar ratio of the monomer and the polymerization initiator, so that the molecular weight of the copolymer can be freely controlled over a wide range by changing the amounts of the monomer and the polymerization initiator used.

[0025] The polymerization initiator used in the polymerization of the copolymer is not particularly limited as long as it causes cationic polymerization to proceed in a living manner. For example, as a living cationic polymerization initiator for vinyl ethers, an HI / I2-based initiator (e.g., JP-A-60-228509) or a polymerization initiator combining a Lewis acid catalyst (organoaluminum compound or the like) with an additive such as a base (ether or the like) (e.g., Japanese Patent No. 3096494, JP-B-7-2805, JP-A-62-257910, JP-A-1-108202 and JP-A-1-108203) is preferably used.

[0026] The amount of the polymerization initiator used is preferably 0.001 to 20 mol %, more preferably 0.01 to 10 mol %, and particularly preferably 1 mol % or less, based on the total amount of the raw material monomers.

[0027] In addition, the living cationic polymerization reaction is preferably carried out in the presence of a suitable organic solvent, but may be carried out in the absence of such an organic solvent. Examples of organic solvents that can be used include aromatic hydrocarbon solvents such as benzene, toluene, and xylene; aliphatic hydrocarbon solvents such as propane, n-butane, isobutane, n-pentane, isopentane, n-hexane, n-heptane, n-octane, isooctane, decane, hexadecane, and cyclohexane; halogenated hydrocarbon solvents such as methylene chloride, ethylene chloride, and carbon tetrachloride; and ether solvents such as diethyl ether, dibutyl ether, tetrahydrofuran (THF), dioxane, and ethylene glycol diethyl ether. These organic solvents may be used alone or in combination of two or more kinds as necessary. Among these organic solvents, aromatic hydrocarbon solvents and aliphatic hydrocarbon solvents are preferred, and toluene or cyclohexane is particularly preferred.

[0028] The polymerization temperature in this polymerization reaction varies depending on the type of polymerization initiator, monomer, and solvent used, but is usually -80 to 150°C, preferably -50 to 100°C, and particularly preferably -20 to 80°C. The polymerization time varies depending on the polymerization initiator, monomer, solvent, reaction temperature, and the like used, but is usually about 10 minutes to 100 hours. The polymerization reaction can be suitably carried out by either a batch method or a continuous method. After the polymerization reaction, if necessary, a purification treatment may be carried out by a known method to remove unreacted monomers.

[0029] <Cancer cell collection filter> The cancer cell collection filter of the present invention (hereinafter, sometimes simply referred to as "filter") has the above-mentioned cancer cell adhesion composition on at least a part of its surface, and by filtering blood with the filter, cancer cells in the blood can be collected. The filter is not particularly limited as long as it is a filter for blood filtration, and any conventionally known filter can be used. The filter only needs to have the cancer cell adhesion composition on at least a part of its surface, but it is preferable that 50% or more of the entire surface area on the side that comes into contact with the blood to be filtered is covered with the cancer cell adhesion composition, more preferably 80% or more, and even more preferably that the entire surface on the side that comes into contact with the blood to be filtered is covered with the cancer cell adhesion composition.

[0030] In terms of more efficient collection of cancer cells, the filter has through-holes with an average pore size of preferably 5 to 30 μm, more preferably 5 to 15 μm, and even more preferably 5 to 10 μm.

[0031] The average aperture ratio of the filter is preferably 5 to 50%, more preferably 10 to 40%, and further preferably 20 to 40%. Here, the average aperture ratio of the filter means the ratio of the area of ​​the through holes to the area of ​​the entire filter.

[0032] As a method for retaining the above-mentioned cancer cell adhesion composition on the surface of a filter, various methods can be used, such as a method of coating the surface of a filter with the cancer cell adhesion composition, a method of bonding the surface of a filter to a biocompatible polymer using active energy rays such as radiation, electron beams, and ultraviolet rays, and a method of reacting and bonding the functional group on the surface of a filter with the cancer cell adhesion composition. When using a coating method, any method such as a coating method, a spray method, or a dip method may be used as a method for coating the cancer cell adhesion composition. Coating is performed by attaching the cancer cell adhesion composition to the surface of a substrate by a dipping method, a spray method, a spin coating method, or the like, and then removing (drying) the solvent. The film thickness after drying is preferably 0.01 μm to 1.0 mm, more preferably 0.1 to 100 μm, and even more preferably 0.5 to 50 μm. If the film thickness is within the above range, the adhesiveness of cancer cells can be fully expressed.

[0033] In order to more firmly fix the composition for cancer cell adhesion to the substrate, the substrate may be heated after being coated with the composition for cancer cell adhesion. The biocompatible polymer may also be crosslinked. For example, a method of crosslinking may include a method of adding a crosslinkable monomer to the polymer material in advance. Electron beams, gamma rays, or light irradiation may be used for crosslinking.

[0034] The material and shape of the filter are not particularly limited, and for example, a porous body, a fiber, a nonwoven fabric, a film, a sheet, or a tube can be used. Materials for the substrate include natural polymers such as cotton and hemp, synthetic polymers such as nylon, polyester, polyacrylonitrile, polyolefin, halogenated polyolefin, polyurethane, polyamide, polysulfone, polyethersulfone, poly(meth)acrylate, halogenated polyolefin ethylene-polyvinyl alcohol copolymer, butadiene-acrylonitrile copolymer, and mixtures thereof. In addition, metals, ceramics, and composite materials thereof can be used, and the substrate may be composed of multiple substrates.

[0035] The metal is not particularly limited, and may be, for example, a precious metal such as gold or silver, a base metal such as copper, aluminum, tungsten, nickel, chromium, or titanium, or an alloy of these metals. The metal may be used alone, or may be used as an alloy with other metals or as a metal oxide to impart functionality. From the viewpoint of cost and availability, it is preferable to use nickel, copper, or a metal containing these as the main component. Here, the main component refers to a component that occupies 50% by weight or more of the material forming the substrate. A screen filter may also be made by forming through holes in these metals using a method such as photolithography.

[0036] <Cancer cell detection method> The method for detecting cancer cells of the present invention comprises a step of filtering blood with the above-mentioned cancer cell-collecting filter, and preferably further comprises a step of culturing the cancer cells collected in the filtering step. Each step will be described in detail below.

[0037] (filtration process) In the method of detecting cancer cells, blood is filtered through a filter, and cancer cells in the blood can be collected with a high recovery rate. For example, a method of filtering blood through a filter includes a method of filtering blood by placing a filter in a flow path through which blood circulates. In such a method, blood pooled in bone marrow, spleen, liver, etc., blood such as umbilical cord blood, lymph, tissue fluid, etc. can be used as samples, but it is most convenient to use peripheral blood circulating in the body. Detecting the presence of circulating cancer cells (hereinafter referred to as "CTC") in peripheral blood is a useful means of determining the progression of cancer.

[0038] When detecting the presence of cancer cells by the method of filtering blood by placing a filter in the flow path through which blood circulates, the method can be carried out, for example, by incorporating a filter in the flow path, introducing peripheral blood into the flow path to capture cells containing CTCs, and confirming whether or not CTCs are present among the captured cells. Examples of methods for introducing blood into the flow path include a method using pressurization from the inlet direction of the flow path, a method using depressurization from the outlet direction of the flow path, and a method using a peristaltic pump. The area of ​​the filter used is, for example, 1 to 10 cm when concentrating CTCs from 1 mL of blood. 2 is appropriate.

[0039] When CTCs are collected by the above method, not only CTCs but also blood cells such as white blood cells are collected at the same time. Therefore, it is necessary to confirm whether the collected cells contain cancer cells. For example, after collecting CTCs by the above method, they can be stained with an antibody against a fluorescently labeled cancer marker to confirm that they are cancer cells. An example of the antibody against the cancer marker is an anti-EpCAM antibody.

[0040] In addition, the genes of the cells collected by the above method can be analyzed to confirm that they are cancer cells. For example, mutations in genes such as p53, K-RAS, H-RAS, N-RAS, BRAF, and APC can be analyzed to confirm that they are cancer cells. The results of these gene analyses can also be used to determine the treatment policy for the patient. Alternatively, the telomerase activity of the cells collected by the above method can be measured to confirm that they are cancer cells.

[0041] (Culture process) The method for detecting cancer cells can detect even a small amount of cancer cells by further including a step of culturing the cancer cells. The method for culturing the cancer cells is not particularly limited, and a method commonly used for cell culture can be used. EXAMPLES

[0042] EXAMPLES Next, the present invention will be described in more detail with reference to examples and comparative examples, but the present invention is not limited to these examples in any way.

[0043] In the examples, the composition ratio of the copolymer is 1 From the results of H NMR analysis, the weight average molecular weight (Mw), number average molecular weight (Mn) and molecular weight distribution (Mw / Mn) were calculated from the results of GPC molecular weight analysis (polystyrene equivalent). The analytical equipment and measurement conditions are as follows:

[0044] (NMR measurement conditions) Equipment: JEOL, Model: AL-400 Solvent: deuterated chloroform ·Measurement temperature: 30℃

[0045] (GPC measurement conditions) Equipment: Tosoh Corporation, "HLC-8320GPC" Detector: RI detector Mobile phase: Tetrahydrofuran ·Flow rate: 1mL / min Column: Showa Denko K.K. "Shodex LF-804" x 3 Column temperature: 40℃

[0046] [Example 1] Synthesis of 2-methoxyethyl vinyl ether / n-butyl vinyl ether random copolymer (MOVE-ran-NBVE) A Schlenk flask equipped with a three-way stopcock was heated and dehydrated at 300°C or higher for 10 minutes under a dry nitrogen atmosphere, and then 76.0 mL of toluene as a solvent, 32.6 mL of ethyl acetate as an added base, 2.3 mL of 2-methoxyethyl vinyl ether (MOVE) as a hydrophilic vinyl ether, 10.3 mL of n-butyl vinyl ether (NBVE) as a hydrophobic vinyl ether, and 494.0 μL of 1-isobutoxyethyl acetate (IBEA) as an initiating species were added and thoroughly stirred.

[0047] Then, the temperature was kept at 0° C., and Et 1.5 AlCl 1.5 0.93M (3.7 mL) was added to initiate polymerization, and the reaction was carried out for 90 minutes.

[0048] The polymerization was terminated with methanol containing a small amount of sodium methoxide (1M). 3 g of ion exchange resin (product name: Amberlyst MSPS2-1·DRY, manufactured by Organo Corporation) was added to the terminated solution and stirred overnight. The solution was then passed through Celite and a filter with a pore size of 1 μm. After evaporating the solvent, the mixture was dried under reduced pressure to obtain the desired random copolymer A. The composition ratio, weight average molecular weight (Mw), number average molecular weight (Mn), and molecular weight distribution (Mw / Mn) of the obtained random copolymer A are shown in Table 1.

[0049] [Example 2] Synthesis of 2-methoxyethyl vinyl ether / n-butyl vinyl ether random copolymer (MOVE-ran-NBVE) Except for changing the amount of IBEA to 185.0 μL, the same procedure as in Example 1 was performed to obtain a random copolymer B. The composition ratio, weight average molecular weight (Mw), number average molecular weight (Mn), and molecular weight distribution (Mw / Mn) of the obtained random copolymer B are shown in Table 1.

[0050] [Example 3] Synthesis of 2-Methoxyethyl Vinyl Ether / n-Butyl Vinyl Ether Random Copolymer (MOVE-ran-NBVE) A random copolymer C was obtained by the same procedure as in Example 1, except that 9.3 mL of MOVE as a hydrophilic vinyl ether, 2.6 mL of NBVE as a hydrophobic vinyl ether, and 300.0 μL of IBEA as an initiating seed were added. The composition ratio, weight average molecular weight (Mw), number average molecular weight (Mn), and molecular weight distribution (Mw / Mn) of the obtained random copolymer C are shown in Table 1.

[0051] [Example 4] Synthesis of 2-Methoxyethyl Vinyl Ether / n-Butyl Vinyl Ether Random Copolymer (MOVE-ran-NBVE) A random copolymer D was obtained by the same procedure as in Example 1, except that 5.8 mL of MOVE as a hydrophilic vinyl ether, 6.4 mL of NBVE as a hydrophobic vinyl ether, and 300.0 μL of IBEA as an initiating seed were added. The composition ratio, weight average molecular weight (Mw), number average molecular weight (Mn), and molecular weight distribution (Mw / Mn) of the obtained random copolymer D are shown in Table 1.

[0052] [Example 5] Synthesis of 2-ethoxyethyl vinyl ether / n-butyl vinyl ether random copolymer (EOVE-ran-NBVE) A random copolymer E was obtained by the same procedure as in Example 1, except that 7.3 mL of 2-ethoxyethyl vinyl ether (EOVE) as a hydrophilic vinyl ether, 6.4 mL of NBVE as a hydrophobic vinyl ether, and 300.0 μL of IBEA as an initiating seed were added. The composition ratio, weight average molecular weight (Mw), number average molecular weight (Mn), and molecular weight distribution (Mw / Mn) of the obtained random copolymer E are shown in Table 1.

[0053] [Example 6] Synthesis of 2-Methoxyethyl Vinyl Ether / 2-Ethylhexyl Vinyl Ether Random Copolymer (MOVE-ran-EHVE) A random copolymer F was obtained by the same procedure as in Example 1, except that 5.8 mL of MOVE as a hydrophilic vinyl ether, 8.0 mL of 2-ethylhexyl vinyl ether (EHVE) as a hydrophobic vinyl ether, and 300.0 μL of IBEA as an initiating seed were added. The composition ratio, weight average molecular weight (Mw), number average molecular weight (Mn), and molecular weight distribution (Mw / Mn) of the obtained random copolymer F are shown in Table 1.

[0054] [Example 7] Synthesis of 2-Methoxyethyl Vinyl Ether / n-Butyl Vinyl Ether Random Copolymer (MOVE-ran-NBVE) Except for changing the amount of IBEA to 74.0 μL, the same procedure as in Example 4 was performed to obtain a random copolymer G. The composition ratio, weight average molecular weight (Mw), number average molecular weight (Mn), and molecular weight distribution (Mw / Mn) of the obtained random copolymer G are shown in Table 1.

[0055] [Comparative Example 1] Synthesis of Poly(n-butyl vinyl ether) (PNBVE) Homopolymer H was obtained by the same procedure as in Example 1, except that no hydrophilic vinyl ether was added, and 12.9 mL of NBVE as a hydrophobic vinyl ether and 300.0 μL of IBEA as an initiating seed were added. The weight average molecular weight (Mw), number average molecular weight (Mn), and molecular weight distribution (Mw / Mn) of the obtained homopolymer H are shown in Table 1.

[0056] [Comparative Example 2] Synthesis of poly(2-methoxyethyl acrylate) (PMEA) I 15.00 g of 2-methoxyethyl acrylate (MEA) as a monomer was dissolved in 60.23 g of 1,4-dioxane, and argon gas was bubbled for 30 minutes. Then, 15.03 mg of 2,2'-azobisisobutyronitrile (AIBN) was added as an initiating species, and polymerization was carried out at 75 ° C for 6 hours while argon gas was bubbled. The polymer solution of the produced polymer was dropped into 1 L of n-hexane to recover the polymer as a precipitate, and the precipitation operation was repeated three times in a THF / n-hexane system to purify it. After the precipitation operation, the substance dissolved in the water was removed by contacting with pure water and stirring for more than 24 hours, and then the solvent was removed by evaporating and vacuum drying (40 ° C for more than 72 hours). Homopolymer I was obtained as the final product. The weight average molecular weight (Mw), number average molecular weight (Mn), and molecular weight distribution (Mw / Mn) of the obtained homopolymer I are shown in Table 1.

[0057] [Comparative Example 3] Synthesis of diethylene glycol monoethyl monovinyl ether / n-butyl vinyl ether random copolymer (EOEOVE-ran-NBVE) In a dry nitrogen atmosphere, a 300 mL three-neck flask equipped with a three-way stopcock was heated and dehydrated at 300°C or higher for 10 minutes, and then 181.0 mL of toluene as a solvent, 76.4 mL of ethyl acetate as an added base, 4.0 mL of diethylene glycol monoethyl monovinyl ether (EOEOVE) as a hydrophilic vinyl ether, 28.2 mL of N-butyl vinyl ether (NBVE) as a hydrophobic vinyl ether, and 4 mM (450.0 μL) of an acetic acid adduct of isobutyl vinyl ether as an initiating species were added and thoroughly stirred.

[0058] Then, the temperature was kept at 0° C., and Et 1.5 AlCl 1.5 Polymerization was initiated by adding 8 mM (8.8 mL), and the reaction was carried out for 90 minutes.

[0059] The polymerization was terminated with methanol containing a small amount of sodium methoxide (1M). 5% by mass of ion exchange resin (product name: Amberlyst MSPS2-1·DRY, manufactured by Organo Corporation) was added to the terminated solution and stirred at room temperature for 1 hour. The solution was then passed through Celite and a filter with a pore size of 1 μm. After evaporating the solvent, the mixture was dried under reduced pressure to obtain the target random copolymer J. The composition ratio, weight average molecular weight (Mw), number average molecular weight (Mn), and molecular weight distribution (Mw / Mn) of the obtained random copolymer J are shown in Table 1.

[0060] [Evaluation test] <Platelet adhesion test> To examine blood compatibility, a platelet adhesion test was carried out on the polyethylene terephthalate (PET) plates (Examples 1 to 7, Comparative Examples 1 to 3) whose surfaces were coated with each of the polymers A to J synthesized above.

[0061] The surface of the PET plate was coated with each polymer by spin coating a 1.0 wt % ethanol solution of each polymer on the surface of the PET plate.

[0062] Human whole blood for experiments purchased from the United States was returned to room temperature before the experiment and centrifuged at 400rcf for 5 minutes in a centrifuge. Approximately 0.5mL of the supernatant was collected and used as platelet rich plasma (PRP). After that, it was further centrifuged at 2500rcf for 10 minutes. Approximately 2mL of the supernatant was collected and used as platelet poor rich plasma (PPP). The seeding density was 4.0×10 7 cells / cm 2Platelet suspensions were prepared by diluting PRP with PPP so that the platelet suspension was 0.2 mL. 0.2 mL of the prepared platelet suspension was dropped with a pipette onto a PET plate with the surface coated with each polymer and onto an uncoated PET plate (blank), and left to stand at 37°C for 60 minutes. The platelets were then rinsed with phosphate buffer solution and incubated in a 1% glutaraldehyde solution at 37°C for 120 minutes to fix the adherent platelets. After that, the platelets were washed with phosphate buffered saline and water, and dried in a container containing silica gel for more than two days. After drying, the sample surfaces were observed under a scanning electron microscope, and a platelet count of 1 × 10 was determined. 5 μm 2 The number of platelets adhering to the area was counted. The number of platelets adhering to an uncoated PET plate (blank) was taken as 1, and the relative number of platelets adhering in each Example and Comparative Example was evaluated according to the following criteria. The evaluation results are shown in Table 1. (Evaluation Criteria) ◎: The number of adhered platelets was less than 0.1. ○: The platelet adhesion number was 0.1 or more and less than 0.2. ×: The number of adhered platelets was 0.2 or more.

[0063] <Cancer cell adhesion test> To investigate the adhesiveness of cancer cells, a cancer cell adhesion test was performed on polyethylene terephthalate (PET) plates (Examples 1 to 7, Comparative Examples 2 and 3) whose surfaces were coated with each of the polymers A to G, I, and J synthesized above. Note that a cancer cell adhesion test was not performed on polymer H because it had a high number of adhered platelets and was poor in blood compatibility.

[0064] In the same manner as in the above-mentioned platelet adhesion test, PET plates coated with each polymer and uncoated PET plates (blanks) were prepared. Each of the prepared PET plates was placed in a 24-well plate (bottom area 1.9 cm 2 ), then the cancer cell suspension (10% serum-supplemented medium at a seeding density of 10,000 cells / cm 21.0 mL of the solution (adjusted to be 1.0 mL) was dropped onto each PET plate in each well with a pipette and left to stand at 37°C for 60 minutes. Human fibrosarcoma cell lines HT-1080 and SW480 were used as cancer cells. Then, the sample was rinsed with phosphate-buffered saline, and the number of cancer cells adhering to the sample surface was counted. To facilitate the counting, the cells were fixed with 4% paraformaldehyde, and the cell nuclei were stained with 4',6-diamino-2-phenylindole (DAPI), and F-actin was stained with phalloidin labeled with Alexa Fluor488. The number of cell nuclei was counted using an all-in-one fluorescence microscope (KEYENCE, BZ-X710), and was taken as the number of cancer cells. The relative number of cancer cells adhering to the uncoated PET plate (blank) was evaluated in each Example and Comparative Example according to the following criteria. The evaluation results are shown in Table 1. Since Polymer J did not show any adhesiveness to HT-1080, an adhesion test to SW480 was not performed. (Evaluation Criteria) ◎: The number of adherent cancer cells was more than 3.0. ○: The number of adherent cancer cells was more than 2.5 and not more than 3.0. △: The number of adherent cancer cells was greater than 1.0 and less than 2.5. ×: The number of adherent cancer cells was 1.0 or less.

[0065] [Table 1]

Claims

1. At least one of the following formula (1): 【Chemistry 1】 (In the formula, R １ indicates a methyl or ethyl group) A repeating unit (A) represented by the formula: At least one of the following formula (2): 【Chemistry 2】 (In the formula, R ２ represents an aliphatic hydrocarbon group) and a repeating unit (B) represented by the following formula (1):

2. The cancer cell adhesion composition according to claim 1 , wherein the biocompatible copolymer is a random copolymer of a segment A consisting of the repeating unit (A) and a segment B consisting of the repeating unit (B).

3. 3. The cancer cell adhesion composition according to claim 1, wherein the composition ratio (molar ratio) of the repeating unit (A) to the repeating unit (B) in the biocompatible copolymer is 90:10 to 10:

90.

4. R in the repeating unit (B) of the biocompatible copolymer ２ is a linear or branched alkyl or alkenyl group having 2 to 10 carbon atoms, or a monocyclic or polycyclic alkyl or alkenyl group having 3 to 20 carbon atoms.

5. R in the repeating unit (B) of the biocompatible copolymer ２ The cancer cell adhesion composition according to any one of claims 1 to 3, wherein is a linear or branched alkyl group having 2 to 10 carbon atoms.

6. The cancer cell adhesion composition according to any one of claims 1 to 5, wherein the ratio of weight average molecular weight (Mw) to number average molecular weight (Mn) of the biocompatible copolymer is 1.0 to 3.

0.

7. The cancer cell adhesion composition according to any one of claims 1 to 6, wherein the number average molecular weight (Mn) of the biocompatible copolymer is 3,000 to 30,000.

8. A cancer cell collecting filter having the cancer cell adhesion composition according to any one of claims 1 to 7 on at least a part of its surface.

9. A method for detecting cancer cells, comprising: A method for detecting cancer cells, comprising a step of filtering blood using the cancer cell collecting filter according to claim 8.

10. The method for detecting cancer cells according to claim 9 , further comprising a step of culturing the cancer cells collected in the filtration step.

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