JP2743480B2 - Antithrombotic polymer material - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to an antithrombotic polymer material.

(Problems to be solved by conventional technology and invention) Conventionally, artificial blood vessels and artificial hearts using polymer materials,
Alternatively, blood processing instruments such as catheters and blood transfusion tubes have been actively used. As is well known, blood coagulates when it comes in contact with foreign matter, and therefore, the polymeric material used in the above various medical devices must be provided with antithrombotic properties.

Conventionally, a method of attaching or coating an anti-blood solid substance such as heparin to the surface of a polymer material has been proposed, but the binding force between the anti-coagulant substance and the polymer material is generally weak, The duration of the effect was not always sufficient.

(Means for Solving the Problems) Accordingly, the present inventors have conducted intensive studies to provide an antithrombotic polymer material having an improved binding force between the anticoagulant substance and the polymer material. By graft-polymerizing an addition-polymerizable ethylenic compound, to which an arginine derivative having antithrombin activity is bonded via its carboxylic acid group, on the surface of a polymer material plasma-treated by glow discharge or the like, Know that the purpose of is achieved,
The present invention has been reached.

That is, the gist of the present invention is to graft polymerize an addition-polymerizable ethylenic compound in which an arginine derivative having antithrombin activity is bonded via a carboxylic acid group of the derivative to a plasma-treated polymer material. An antithrombotic polymer material characterized by being obtained by:

Hereinafter, the present invention will be described. The antithrombotic component in the antithrombotic polymer material of the present invention is an arginine derivative having at least one carboxylic acid group and having antithrombin activity. JP-A-52-97934, JP-A-5-97934
No. 5-33499, European Patent No. 8746, Journal of Medical Chemistry (J. Med. Ch.)
em.) 23, from 1,293 to 1,299 (1980), or, Biochemistry (Biochemistry) 23, include compounds such as those described in 85-90 (1984). Among them, an arginine derivative represented by the following general formula (I) is preferable.

[In the above formula, R ¹ is (Wherein, R ³ represents an alkyl group, an alkoxyalkyl group or a tetrahydrofurfuryl group of C ₂ -C ₇ of C ₁ ~C _6, R ⁴
Represents an alkylene group of C ₁ -C _3. ) Or (Wherein, R ⁵ represents a hydrogen atom or a C ₁ -C ₄ alkyl group), and R ² represents (Wherein, R ⁶ and R ⁷ represent a hydrogen atom, a C ₁ -C ₄ alkoxy group or a C ₁ -C ₄ alkylamino group) or (In the formula, R ⁸ represents a hydrogen atom or a C _{1 to} C ₄ alkyl group.), And m represents an integer of 3 to 4. In the above formula, R ¹ is Or Represents a group represented by R ³ is a C ₁ to C ₃ group such as a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group, and a hexyl group.
Alkyl C _6, represents a methoxymethyl group, methoxyethyl group, ethoxymethyl group, ethoxyethyl group, a butoxymethyl group, alkoxyalkyl group or a tetrahydrofurfuryl group of C ₂ -C ₇ such as butoxyethyl, R ⁴ methylene group, ethylene group, an alkylene group of C ₁ -C ₃ such as a propylene group, R ⁵ is a hydrogen atom or a methyl group, an ethyl group, a propyl group, an alkyl group of C ₁ -C _4, such as butyl Represent.

Represents a group represented by R ⁶ and R ^{7 each} represent a hydrogen atom, a methoxy group, an ethoxy group, a propoxy group, a C ₁ to
Alkoxy or methylamino group of C _4, ethylamino group, propylamino group, butylamino group, dimethylamino group, diethylamino group, methylethylamino group, an alkylamino group of C ₁ -C _4, such as methyl propylamino group R ⁸ represents a hydrogen atom or a C ₁ -C ₄ alkyl group such as a methyl group, an ethyl group, a propyl group, and a butyl group. m is 3 ~
Represents an integer of 4. Specific examples of the arginine derivative include, for example, compounds shown in Table 1 below.

In the present invention, the arginine derivative is graft-polymerized to a polymer material with an addition-polymerizable ethylenic compound bonded via its carboxylic acid group.

As the ethylenic compound, any compound having an ethylenically unsaturated group capable of undergoing graft polymerization and a group capable of binding to the carboxylic acid group of the arginine derivative, for example, an amino group, may be used.

For example, compounds obtained by reacting unsaturated carboxylic acid halides such as acrylic acid and methacrylic acid with diamines such as H ₂ N— (CH ₂ ) _n —NH ₂ (n = 1 to 10) are exemplified. Can be

Although there is no particular limitation on the polymer material, for example, polyurethane, polyamide, polypropylene, polyvinyl chloride, polyethylene, polyvinylidene chloride, polystyrene, polycarbonate, polyurea, polyester and the like can be used.

In the graft polymerization of an addition-polymerizable ethylenic compound having the arginine derivative onto a polymer material, first,
Plasma treatment is performed on the surface of the polymer material by glow discharge or corona discharge. 0.1-0.3 Torr for glow discharge,
The treatment is performed under the conditions of 0.1 to 10 mA for about 5 seconds to 3 minutes. In the case of corona discharge, the maximum applied voltage is 10 to 20 kV in the atmosphere.
The processing is performed under the condition of about 2 to 3 minutes.

Next, the polymer material subjected to the plasma treatment is immersed in a 5-80% aqueous solution of an addition-polymerizable ethylenic compound having the arginine derivative, and is subjected to UV irradiation or heat treatment to be graft-polymerized on the surface of the polymer material.

UV irradiation is performed by a mercury lamp at 4 to 30 ° C. for 6 to 24 hours, and heat treatment is performed at 60 to 130 ° C. for 3 to 12 hours.
Perform under time conditions. At this time, the graft amount of the addition-polymerizable ethylenic compound having the arginine derivative is 0.
It is good to choose so as to be in the range of 1 to 10 mg / cm ² .

Thus, the antithrombotic polymer material of the present invention can be obtained.

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

Example 1 (1) 15 ml of a 1.0 mmol solution of acrylic chloride in ethyl acetate cooled to −78 ° C. was gradually added dropwise to a solution of 2.0 mmol of ethylenediamine in ethyl acetate at −4 ° C. with vigorous stirring.
After reacting at -4 ° C for 1 hour, the reaction was further performed at room temperature for 4 hours. After completion of the reaction, the solvent was removed, and ethyl acetate 10
0 ml was added and the product was extracted. After passing the precipitate at a temperature of 55 to 65 ° C, ethyl acetate was distilled off under reduced pressure at 30 to 40 ° C,
Vacuum drying was performed at room temperature for 12 hours to obtain 0.8 g of N-aminoethylene acrylamide as a white powder.

Then, at 0 ° C., 0.10 mmol of (2R, 4R) -4-methyl-1- [N ² -((RS) -3-methyl-1,2,3,4-tetrahydro-8-quinolinesulfonyl)- L-arginyl]
-2-piperidinecarboxylic acid (hereinafter referred to as MD-805), 0.11 mmol of dicyclohexylcarbodiimide and
DM containing 0.15 mmol of hydroxybenzotriazole
To 20 ml of the F solution, 20 ml of 0.1 mmol of N-aminoethyleneacrylamide obtained above was added, and the mixture was reacted at room temperature for 72 hours with stirring. At the end of the reaction, 60 ml of ethyl acetate was added, followed by washing three times with 4% sodium bicarbonate, 10% citric acid and saturated saline sequentially, and then ethyl acetate was distilled off. The resulting residue was subjected to silica gel chromatography. It was separated and purified by chromatography [developing solvent: ethyl acetate / chloroform = 3/1], recrystallized from ethyl acetate / ethyl ether = 10/1, dried in vacuo, and dried with N-aminoethylene acrylamide.
A reaction product of MD-805 (hereinafter referred to as CM-MD-805) was obtained. The yield was 63.5%. Figure 1 shows CM-MD-805 and MD
The IR spectrum of -805 was shown. The results of elemental analysis were as follows.

Calculated C: 55.1%, H: 7.28%, N: 18.54% Analytical C: 55.76%, H: 7.02%, N: 18.72% Instead of MD-805, t-butoxycarbonylarginine is used as a control. N'-t-
Butoxycarbonylamide-N-aminoethyleneacrylamide (hereinafter referred to as CM-Arg) was obtained.

 The results of the elemental analysis were as follows.

Calculated value C: 51.89%, H: 8.11%, N: 22.70% Analysis value C: 51.74%, H: 8.32%, N: 22.54% (2) Journal of Biomedical Materials Research (J. Biomed. Mater. Res. ), 20,1157 (198
Glow discharge treatment of a polyurethane (polytetramethylene glycol: 4,4'-diphenylmethane diisocyanate: butanediol = 1: 2: 1 (charge ratio)) film (1.5 cm × 1.5 cm) synthesized as described in 6). (Condition: 0.2Tor
r, 5 mA, 1 minute), and put it in an aqueous solution of (a) 5%, (b) 10%, (c) 15% and (d) 20% of CM-MD-805 obtained in (1) above. UV irradiation was performed at 20 ° C. for 24 hours in a nitrogen atmosphere to graft-polymerize CM-MD-805 on the surface of the polyurethane film. After the completion of the reaction, the resultant was washed three times with distilled water and dried at room temperature. The obtained product is hereinafter referred to as GR-PEU-MD
(A, b, c, d).

GR-PEU-AR (a, b, c, d) was synthesized in the same manner except that CM-Arg was used instead of CM-MD-805.

CM-MD-805 and CM-Arg for each polyurethane film
The amount of the graft was measured as follows. That is,
Each polyurethane film was placed in 2 ml of 6N hydrochloric acid and subjected to a hydrolysis reaction at 110 ° C. for 12 hours. After the reaction, the reaction was neutralized with 6N hydrochloric acid and sodium hydroxide, and a borate buffer (pH 8.5) was added to a total volume of 10 m.
l. Fluram (25 mg fluram / 100 ml acetone) was added thereto, mixed immediately, excited at 390 mm, and the content was quantified by the fluorescence emission intensity at 480 nm. Figure 2 shows the result.
It was shown to. In addition, it measured using the polyurethane film (henceforth PEU) without a graft polymerization as a control.

Test Example 1 (a) Production rate of thrombus on GR-PEU-MD membrane surface 200 μl of canine citrated whole blood was dropped on the polymer membrane, and 20 μl of 0.1 M calcium chloride solution was added.
The mixture was allowed to stand for 1 minute, the formed thrombus was fixed with formalin, dried, and weighed to measure.

The amount of thrombus generated when the above experiment was performed on a glass plate was set to 100%, and the GR-PEU-MD (a, b, c,
The thrombus formation rate of d) was determined. The results are shown in Table 2.
For comparison, PEU and GR-PEU-AR obtained in Example 1 were used.
The results when (a, b, c, d) were used are also shown.

(B) Adhesion of platelets on GR-PEU-MD membrane surface Platelet-rich plasma (PRP) of citrated dogs
It was dropped on the surface of GR-PEU-MD (d). After immersion at 37 ° C for 20 minutes, it was washed twice with physiological saline solution (PBS),
The cells were fixed in PBS containing 2.5% glutaraldehyde for 20 hours. Next, after dehydration treatment with ethanol and drying, the surface of the membrane was observed with an SEM to measure the amount of platelet adhesion to the membrane surface. Table 3 shows the results. For comparison, the results when PEU and GR-PEU-AR (d) were used are also shown.

(Effect of the Invention) The antithrombotic polymer material of the present invention can be introduced into the polymer material more since the antithrombotic component has a relatively low molecular weight,
In addition, since it is graft-polymerized, the bond with the polymer material is better than before, and the effect can be maintained for a long time.
It can be advantageously used for various blood processing instruments.

[Brief description of the drawings]

FIG. 1 is a diagram showing the IR spectra of MD-805 and CM-MD-805, wherein (A) is the IR spectrum of MD-805 and (B) is the CM-MD.
The IR spectrum of -805 indicates absorption indicating allyl, absorption indicating secondary amide, and absorption indicating sulfonamide. The vertical axis indicates absorption, and the horizontal axis indicates wavelength (cm ^-1 ). FIG. 2 shows GR-PEU-MD (a, b, c, d) obtained in Example 1 (in the figure,
(Shown by △) and GR-PEU-AR (a, b, c, d) (in the figure, ○
CM-MD-805)
FIG. 6 is a diagram showing the amount of CM-Arg. The vertical axis indicates the content of the graft-polymerized CM-MD-805 or CM-Arg (× 10 ⁻⁶ mo
l / cm ² ), and the horizontal axis indicates the content (%) of CM-MD-805 or CM-Arg.

Claims (1)

(57) [Claims] 1. An addition-polymerizable ethylenic compound in which an arginine derivative having antithrombin activity is bonded via a carboxylic acid group of the derivative to a plasma-treated polymer material, which is obtained by graft polymerization. An antithrombotic polymer material, characterized in that: