JP2846390B2 - Light-transmitting aromatic vinyl polymer molded article having amino groups on the surface and method for producing the same - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to an aromatic vinyl polymer molded article suitable for immobilizing a physiologically active substance and a method for producing the same.

(Prior art) Molded articles with a physiologically active substance immobilized on the surface are widely used as cell culture equipment, blood purification equipment, etc.
It is an important substance that is expected to expand further in the future.

Aromatic vinyl polymers represented by polystyrene are one of the most widely used plastics because of their excellent light transmittance, mechanical properties, processability, and economy. However, it has the drawback of being hydrophobic, which makes it unsuitable for use in direct contact with cells such as syringes, blood circuits, and cell culture dishes. Further, it does not have an amino group or a carboxyl group necessary for immobilizing a physiologically active substance. As a method for improving this drawback, a plasma discharge treatment is currently performed. This plasma discharge treatment method is a method in which a functional group is shallowly inserted into the surface of a plastic, and is a simple and convenient method in which a functional group can be inserted without changing the physical properties of the whole molded article. However, although this method can be applied to a molded article having a simple shape such as a cell culture dish, it has a drawback that it cannot be applied to the inside of a molded article having a complicated shape, for example, the inner surface of a tube or a bottle. Further, since the chemical structure of the introduced functional group is unclear, its application is limited.

On the other hand, chloroacetamidomethylated polystyrene fiber (Japanese Patent Application Laid-Open No. 57-12008) is known as a product obtained by introducing a functional group into a polystyrene molded product. The shape of the fiber is maintained. Therefore, this type of molded product is limited to a fibrous product, and its use is narrow. Also, since it is no longer polystyrene, the light transmittance is lost.

Further, when the reaction method used here is applied to a molded article having a large thickness such as a petri dish, there is a problem in a method of stopping the reaction. For example, after the petri dish is brought into contact with the reaction solution, when the molded article is immersed in a large excess of cold water, the nitrobenzene used as the solvent will undergo phase separation, dissolving or devitrifying the unreacted polystyrene. There is a disadvantage.

(Problems to be Solved by the Invention) In view of the problems of the related art, the present inventors have made various complicated shapes such as a syringe, a blood circuit, and a cell culture dish, and have a reactive surface on the surface. As a result of intensive studies to find a molded article of an aromatic vinyl polymer into which a functional group, particularly a primary amino group has been introduced, and a method for producing the same, the present invention has been achieved.

(Means for Solving the Problems) The configuration of the present invention for achieving such problems includes the following technical means.

(1) Part or all of the surface of an aromatic vinyl polymer molded article comprising a repeating unit represented by the following general formula (I): (In the formula, R ₁ represents a hydrogen atom or a methyl group, and A ₁ represents a benzene ring.) It is composed of an aromatic vinyl polymer represented by the following general formula (II), and is perpendicular to the film surface. Absorbance of 500-700
A light-transmitting aromatic vinyl polymer molded article having an amino group on the surface formed and coated with a film having a wavelength of 3 or less for light having a wavelength of nanometer.

(Wherein, R ₁ represents a hydrogen atom or a methyl group, A _{1 represents} a benzene ring, A ₂ represents a 1,4-disubstituted benzene ring. Further, n and m are such that n / (n + m) is 0.01 or more. (2) A molded article of the aromatic vinyl polymer represented by the general formula (I) is converted to an N-methylol-α-haloacylamide represented by the following general formula (III) by mononitration. After contacting with a solution comprising a hydrocarbon and sulfuric acid, the surface is washed with a lower alcohol or a lower fatty acid, and then treated with a hydrophilic tertiary amine and water. A process for producing a molded article of a light-transmitting aromatic vinyl polymer having a group.

(Wherein, R ₂ represents a hydrogen atom or a lower alkyl group;
Represents a halogen atom. (3) Molded products are petri dishes, test tubes, tubes, films, cell culture instruments, extracorporeal circulation columns, blood storage containers, bottles,
The molded article of a light-transmitting aromatic vinyl-based polymer having an amino group on its surface according to the above (1), which is a kind selected from a syringe and a catheter.

The aromatic vinyl polymer molded product represented by the general formula (I) in the present invention means a molded product of a homopolymer of styrene or α-methylstyrene or a copolymer containing these as a main component. There is no particular limitation on the thickness of the molded product.
Those having a thickness of 0 micron or more are preferably used. Specific examples thereof include petri dishes, examiners, tubes, films, bottles, syringes, catheters, and the like.

The aromatic vinyl polymer film represented by the general formula (II) referred to in the present invention is a polymer film composed of the aromatic vinyl polymer represented by the general formula (II), and A film having sufficient light transmission to allow the shape of the material present on the opposite side to be ascertained. The light transmittance of a film is determined mainly by scattering on the film surface and absorption of the film.
It is necessary that the absorbance for light having a wavelength of about 700 nanometers is 3 or less. The smaller the absorbance, the better the light transmittance of the molded article. It is more preferable that the absorbance is 1 or less.

The chemical structure of the polymer film of the present invention is not particularly limited as long as it is represented by the general formula (II), and n / (n + m)
If the particle size is too small, quantification becomes difficult and the structure becomes uncertain. Therefore, a particle size of usually 0.01 or more, more preferably 0.1 or more is used.

Specific examples of the light-transmitting aromatic vinyl-based polymer molded product having an amino group on the surface formed and coated with the aromatic vinyl-based polymer film represented by the general formula (II) of the present invention include:
A molded article of an aromatic vinyl polymer represented by the general formula (I) obtained by chemically treating the surface of the molded article to form an aromatic vinyl polymer represented by the general formula (II) to a certain depth, A molded article of an aromatic vinyl polymer represented by I) coated with a liquid aromatic vinyl polymer represented by the general formula (II), and an aromatic vinyl polymer represented by the general formula (I): Examples include a product obtained by laminating a film-like aromatic vinyl polymer represented by the general formula (II) on the surface of a united molded product. In particular, the molded product has discontinuous chemical structure in the depth direction. Since it does not have, it has essentially good peeling resistance, and is particularly preferable.

If the thickness of the film is too large, the light transmittance is lowered and the rub resistance is also lowered. Particularly preferred is 50 microns or less, especially 5 microns or less.

In the production of the molded article of the present invention, a necessary part of the aromatic vinyl polymer molded article represented by the general formula (I) described in claim 1 is replaced with an N-methylol-based compound represented by the general formula (III).
It is immersed in a solution composed of α-haloacylamide, mononitrated hydrocarbon and sulfuric acid or a solution obtained by further adding paraformaldehyde to this solution for a required time, washed with a lower alcohol or a lower fatty acid, and then washed with a hydrophilic alcohol. It is easily achieved by treatment with a tertiary amine and water. In this case, the film thickness and chemical structure general formula (II)
N / (n + m) can be adjusted by selecting the composition of the reaction solution of N-methylol-α-haloacylamide and the reaction temperature and time.

The N-methylol-α-haloacylamide used in the present invention is not particularly limited as long as it is represented by the general formula (II). Specific examples thereof include N-methylol-α-chloroacetamide, N-methylol-α-bromoacetamide, N-methylol-α-iodoacetamide, N-methylol-α-chloropropionamide,
N-methylol-α-bromo-n-butyric acid amide and N-methylol-α-bromo-n-valeric acid amide acid are exemplified. ₁ is a hydrogen atom, X is a chlorine atom,
That is, N-methylol-α-chloroacetamide is particularly preferably used. Further, as the mononitrated hydrocarbon, 1-nitropropane, 2-nitropropane, nitrobutane, nitromethane, nitrobenzene, and the like are preferably used. Nitropropane, which is easy to operate, is particularly preferably used.

There is no particular limitation on the amount of N-methylol-α-haloacylamide used, but the smaller the amount, the easier the transparency of the molded article is to be maintained. Since it does not dissolve, usually 0.5 to
It is used at a concentration of 8%, especially 1-8%. Further, in order to treat the molded article without damaging it, it is important to use the reaction solution as a homogeneous solution.

The concentration of the mononitrated hydrocarbon is preferably used within a range of 70% or less so that the reaction solution becomes a homogeneous solution. The higher the concentration of the mononitrated hydrocarbon, the greater the swelling of the molded article, and the higher the reaction rate.

The preparation temperature of the haloacylamide methylation reaction solution is 5 to 10 ° C below zero in order to avoid decomposition of the N-methylol compound.
The contact temperature between the haloacylamide methylation reaction solution and the molded article is preferably 0 ° C. to avoid side reactions.
To 20 ° C.

The depth and amount of the α-haloacylamidomethyl group introduced into the molded article depend, of course, on the reaction temperature and the composition of the reaction solution, especially the concentration of the N-methylol compound.

The lower alcohol and lower fatty acid used for washing the molded product treated with the reaction solution of N-methylol haloacylamide of the present invention may be anything as long as it is not decomposed by sulfuric acid and is mixed with sulfuric acid and mononitrated hydrocarbon.
There is no particular limitation. Specific examples thereof include lower alcohols such as methanol, ethanol, isopropanol, n-butanol and methyl cellosolve, and lower fatty acids such as formic acid, acetic acid and propionic acid. In addition, it is preferable to use these materials at a lower use temperature because a temperature increase due to the solvation heat of sulfuric acid can be avoided and, therefore, decomposition of the chloroacetamidomethyl group can be avoided. Usually below zero 30
Temperatures in the range C to 20C are preferably used.

The hydrophilic tertiary amine used in the present invention is not particularly limited as long as it is a hydrophilic tertiary amine. Specific examples thereof include N, N-dimethylaminoethanol, N, N-
Dimethylamino-2-propanol, N, N-dimethylamino-3-propanol, N, N-dimethylaminoethanol, N, N-dimethylamino-2-butanol,
Trimethylamine and the like can be mentioned, but N, N-dimethylaminoethanol is particularly excellent from the viewpoint of high reactivity and economy. A solvent is not required, but any solvent that does not dissolve polystyrene, does not dissolve tertiary amines, and is neutral can be used. Suitable solvents are lower alcohols such as methanol, ethanol, isopropanol, n-butanol and methyl cellosolve, and dimethyl sulfoxide.

As the hydrolysis treatment with the hydrophilic tertiary amine and water performed in the present invention, the molded article having the surface into which the haloacetamidomethyl group is introduced may be warmed with an aqueous solution, or may be subjected to the hydrophilic tertiary amine.
This is achieved by treating with a solution of a secondary amine and then immersing in water. At this time, 70 ° C to prevent deformation and devitrification of the molded product
It is important to do at the following temperatures:

Aminomethyl groups present on the surface of the molded article of the present invention,
Not only does it help to improve the cell affinity of the molded product, but it can also be used directly for immobilizing a physiologically active substance by utilizing its reactivity.

Examples of the use of the molded article of the present invention include immobilizing a physiologically active substance such as an antithrombotic agent, an antibiotic, or an antibody on the surface of the molded article to form a blood circuit, a catheter, a syringe, a cell culture instrument, or an extracorporeal circulation. It can be used for columns, blood storage containers and the like.

(Effect of the Invention) The molded article of the present invention has a clear surface functional group, so that a physiologically active substance can be immobilized with a clear chemical bond, and the aromatic vinyl polymer moiety represented by the general formula (I) Can be easily extracted and removed with a solvent, so that the immobilization density and chemical structure can be clearly analyzed after immobilization, and the mechanical properties of polystyrene are maintained as a whole molded product. Since it is possible and has light transmittance, it has an advantage that the inside can be visually recognized at the time of use.

Further, the usefulness of the method for producing a molded article of the present invention is that the molded article of any shape can be produced, inexpensive polystyrene can be used, and there is no discontinuity in the chemical structure, so the peeling resistance is excellent. It has advantages such as being able to manufacture molded articles.

Hereinafter, the present invention will be described more specifically with reference to examples.

 In addition, the evaluation method in an Example followed the following.

1. Chemical analysis of the aromatic vinyl polymer film represented by the general formula (II). Soxhlet extraction of the molded product with methylene chloride yields an insoluble film, which is dried in vacuum, weighed, and absorbed by infrared light. Spectrum measurement and the like were performed.

2. Infrared absorption spectrum KB using Shimadzu Fourier transform infrared spectrophotometer FTIR-4300
r Measured by the tablet method.

3. Measurement of light transmittance of molded product The absorbance in the vertical direction of the plate-shaped molded product was measured at 500 nanometers using a Shimadzu spectrophotometer UV2100. Measuring the absorbance of the same thickness of the polystyrene plate molded as a control, under both was A _500. Therefore, the smaller the value, the higher the light transmittance of the molded product.

Example 1. A mixed solution of 500 ml of 1-nitropropane and 272 ml of sulfuric acid was added to 0
After cooling to 2 ° C., 20.4 g of N-methylol-α-chloroacetamide was added and dissolved at 0 to 10 ° C.

After the temperature of this solution was raised to 10 ° C., 9 ml each of the solution was placed in 50 polystyrene culture dishes (thickness: 1 mm) of φ3.5 cm × 1.2 cmH, and reacted at room temperature for 1 hr. The reaction solution was discarded, and the culture dish was immersed in methanol at a temperature of 20 ° C. below zero, further washed with methanol and water, and dried under vacuum to obtain an intermediate molded product A in which only the inner surface was chloroacetamidomethylated. A _{500 of} this molded product was 0.050.

When one molded product A obtained as described above was subjected to Soxhlet extraction with methylene chloride, 3.1 mg of a thin film-like insoluble matter was obtained. Since the surface area of the reaction surface of the culture dish is 21.1 cm ² , the thickness of the reaction product is considered to be approximately 1.5 μ. The degree within this infrared absorption spectrum of the insoluble matter, 1659 cm ^-1 (amide -I) and 1529cm ^-1 (amide -II) to the primary amide strong absorption and amide N-H group to 3297cm ^-1 of group Was confirmed, and its structure was confirmed.

The intermediate molded product A40 obtained above was immersed in 2000 ml of 10% dimethylaminoethanol / dimethyl sulfoxide solution, allowed to stand at room temperature for 18 hours, and then immersed in 2000 ml of 10% dimethylaminoethanol aqueous solution at 70 ° C. Heated for 2 hr. The molded product was washed and dried to obtain a molded product B of the present invention. A _{500 of} this molded product was 0.050.

When the above-mentioned molded product B of the present invention was subjected to Soxhlet extraction with methylene chloride, 2.3 mg of a thin film-like insoluble matter was obtained. In the infrared absorption spectrum of this insoluble matter, the molded product A
1659 cm ^-1 (amide-I) and 1529 cm
The structure was confirmed because strong absorption cm ^-1 of the primary amide group of ^-1 (amide-II) had completely disappeared.

The molded article B of the present invention was immersed in 200 ml of water, and N / 100−
After adjusting the pH to 5 to 6 with hydrochloric acid, 5 ml of a 0.2 mg / ml sodium glucuronate aqueous solution and 8 mg of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride were added, and
It was left at room temperature for 48 hours. The concentration of glucuronic acid in the mother liquor was analyzed by the phenol-sulfuric acid method and found to be 0.122 mg / ml, indicating that 0.391 mg was immobilized. Also,
It was confirmed that glucuronic acid on the molded product was not eluted even after washing with N / 40-NaOH.

Example 2 A mixed solution of 148 ml of 1-nitropropane and 81 ml of sulfuric acid was diluted with 0
After cooling to 0 ° C, 3 g of N-methylol-α-chloroacetamide was added and dissolved at 0 to 10 ° C.

After the temperature of the solution was raised to 10 ° C., 35 ml of each solution was placed in six polystyrene culture dishes (diameter: 1 mm) of φ8.6 cm × 1.2 cmH, and reacted at room temperature for 1 hour. The reaction solution was discarded, and the culture dish was immersed in methanol at a temperature of 20 ° C. below zero, further washed with methanol and water, and dried in vacuo to obtain an intermediate molded product C in which only the inner surface was chloroacetamidomethylated.

When one molded product C obtained above was subjected to Soxhlet extraction with methylene chloride, 6.5 mg of a thin film of insoluble matter was obtained.
Since the surface area of the reaction surface of the culture dish is 74.3 cm ² , the thickness of the reaction product is considered to be 0.87 μm. The degree within this infrared absorption spectrum of the insoluble matter, 1659 cm ^-1 (amide -I) and 1529cm ^-1 (amide -II) to the primary amide strong absorption and amide N-H group to 3297cm ^-1 of group Was confirmed, and its structure was confirmed.

The molded product C5 obtained above was immersed in 1000 ml of a 10% dimethylaminoethanol / dimethyl sulfoxide solution,
After standing at room temperature for 18 hours, it was heated at 70 ° C. for 2 hours in 2000 ml of a 10% aqueous solution of dimethylaminoethanol. The molded article was washed with water and dried to obtain a molded article D of the present invention. Of this molded product
A ₅₀₀ was 0.036.

When one molded article D of the present invention was subjected to Soxhlet extraction with ethylene chloride, 5 mg of a thin film of insoluble matter was obtained.
In the infrared absorption spectrum of this insoluble matter, the strong absorption cm ^-1 of the primary amide group at 1659 cm ^-1 (amide-I) and 1529 cm ^-1 (amide-II) observed in the molded product C completely disappeared. The structure was confirmed.

Example 3 A mixed solution of 98.3 g of 1-nitropropane and 53.4 g of sulfuric acid was mixed with 0
After cooling to 0 ° C, 7.0 g of N-methylol-α-chloroacetamide was added and dissolved at 0 to 10 ° C.

After the temperature of the solution was raised to 10 ° C., 9 ml each of the solution was placed in 26 polystyrene culture dishes (1 mm thick) of φ3.5 cm × 1.2 cmH, and reacted at room temperature for 1 hour. The reaction solution was discarded, and the culture dish was immersed in methanol at 20 ° C. below zero, further washed with methanol and water, and dried under vacuum to obtain an intermediate molded product E in which only the inner surface was chloroacetamidomethylated. The E _{500 of} this molded product was 0.940.

When one molded product A obtained above was subjected to Soxhlet extraction with methylene chloride, 7.0 mg of a thin film-like insoluble matter was obtained. Since the surface area of the reaction surface of the culture dish is 21.1 cm ² , the thickness of the reaction product is considered to be approximately 3.3 μm. The structure of the insoluble matter was confirmed by an infrared absorption spectrum.

The intermediate product E20 obtained above was immersed in 2000 ml of a 10% dimethylaminoethanol / dimethyl sulfoxide solution, allowed to stand at room temperature for 18 hours, and then placed in a 2000 ml 10% dimethylaminoethanol aqueous solution at 70 ° C. Heated for 2 hr. The molded product was washed with water and dried to obtain a molded product F of the present invention. The A _{500 of} this molded product was 0.490.

When one molded article F of the present invention was subjected to Soxhlet extraction with methylene chloride, 2.3 mg of a thin film-like insoluble matter was obtained. In the infrared absorption spectrum of this insoluble matter, the molded product E
Accepted 1659 cm ^-1 (amide -I) and 1529cm ^-1 primary amide groups strongly absorb cm ^-1 of (amide -II) confirms the structure since it had completely disappeared.

Example 4 5 ml and 2.5 ml of the reaction solution of Example 1 were placed in each hole of a multi-plate for cell culture made of polystyrene and 12F and 24F (manufactured by Sumitomo Bakelite), and reacted at room temperature for 2 to 60 minutes. The reaction solution was discarded, poured into methanol at a temperature of 20 ° C. below zero, and treated in the same manner as in Example 1 to obtain an intermediate molded product in which only the inner surface was chloroacetamidomethylated. This was treated with a 10% dimethylaminoethanol / dimethylsulfoxide solution and a 10% dimethylaminoethanol aqueous solution in the same manner as in Example 1 to obtain a molded article of the present invention. The A _{500 of} this molded product was 0.490.

Claims (3)

(57) [Claims] 1. A part or all of the surface of an aromatic vinyl polymer molded article comprising a repeating unit represented by the following general formula (I): (In the formula, R ₁ represents a hydrogen atom or a methyl group, and A ₁ represents a benzene ring.) It is composed of an aromatic vinyl polymer represented by the following general formula (II), and is perpendicular to the film surface. Absorbance of 500-700
A light-transmitting aromatic vinyl polymer molded article having an amino group on the surface formed and coated with a film having a wavelength of 3 or less for light having a wavelength of nanometer. (Wherein, R ₁ represents a hydrogen atom or a methyl group, A _{1 represents} a benzene ring, A ₂ represents a 1,4-disubstituted benzene ring. Further, n and m are such that n / (n + m) is 0.01 or more. Is a natural number.) 2. An aromatic vinyl polymer molded product represented by the general formula (I) is prepared from an N-methyl roll-α-haloacylamide, a mononitrated hydrocarbon and sulfuric acid represented by the following general formula (III): (1) washing with a lower alcohol or a lower fatty acid, followed by treatment with a hydrophilic tertiary amine and water.
The method for producing a molded article of a light-transmitting aromatic vinyl polymer having an amino group on the surface according to the above item. (Wherein, R ₂ represents a hydrogen atom or a lower alkyl group;
Represents a halogen atom. ) 3. The molded article is a kind selected from a petri dish, a test tube, a tube, a film, a cell culture device, a column for extracorporeal circulation, a blood storage container, a bottle, a syringe, and a catheter. The molded article of a light-transmitting aromatic vinyl polymer having an amino group on the surface according to claim (1).