JP2008194362A - Antithrombogenic coating agent and medical device - Google Patents

Abstract

A phosphorcholine group-containing antithrombotic coating agent having a temperature responsiveness that is hydrophilic at a temperature lower than a predetermined temperature (T) lower than a living body temperature and becomes hydrophobic at a temperature higher than the predetermined temperature. I will provide a.
An antithrombogenic coating agent comprising a polymer material having a phosphorcholine group-containing polymer chain and a polymer chain of N-isopropylacrylamide or a derivative thereof. The polymer material is hydrophilic at a temperature lower than a predetermined temperature (T) lower than the living body temperature, and is hydrophobic at a temperature higher than the predetermined temperature (T). This polymer material preferably uses a compound having three or more N, N-dialkyl-dithiocarbamylmethyl groups in the same molecule as an iniferter, and 2- (meth) acryloyloxyethyl-2 ′-(trimethylammoni E) Block polymerization of ethyl phosphate and N-isopropylacrylamide.
[Selection figure] None

Description

  The present invention can maintain excellent anticoagulability (antithrombogenicity) over a long period of time, and can be applied to living tissue or as a constituent material of various medical devices such as artificial organs and artificial blood vessels. More particularly, the present invention relates to an antithrombotic coating agent having a phosphorcholine group. The present invention also relates to a medical device coated with this antithrombotic coating agent.

  I. When blood comes into contact with foreign substances, it has the property of coagulating due to the action of various components in the blood. Therefore, constituent materials for medical devices used for parts that come into contact with blood, such as artificial hearts, artificial heart valves, artificial blood vessels, vascular catheters, cannulas, cardiopulmonary bypasses, vascular bypass tubes, aortic balloon pumping, transfusion devices and extracorporeal circulation circuits Is required to have high anticoagulability. However, many of the constituent materials of conventional medical devices are inevitably caused by blood coagulation when used for a long period of time, which is not sufficient in terms of anti-blood coagulation sustainability.

  II. It is known that a phosphoroline group-containing monomer polymer has antithrombotic properties as described in paragraph 0002 of JP-A-9-183819. In the same issue, random copolymers or alternating copolymers of phosphorylcholine group-containing monomers such as 2- (meth) acryloyloxyethyl-2 ′-(trimethylammonio) ethyl phosphate and silyl group-containing monomers and medical treatment thereof A medical device coated on the surface of the material is described.

JP-A-9-183819

The copolymer of the above-mentioned JP-A-9-183819 adheres to the material surface by hydrolysis of its silyl group. (Paragraphs 0052-0053 of the same publication).
Therefore, when the copolymer of the same number is used as an aqueous solution for coating, as described in paragraph 0052, it is necessary to dissolve the copolymer in water when coating, and it cannot be stored for a long time as an aqueous solution for coating.

  In the case where the copolymer of the same number is stored as a stable solution for a long period of time, it is necessary to use an alcohol solution as seen in the 0053 paragraph of the same number or the example (paragraph 0091).

  Thus, when alcohol is used as a solvent, it may not be used for surface treatment of cells used in regenerative medicine and hybrid tissues composed of synthetic or biological materials, or biological tissues extracted from a host (cells may be affected by alcohol). To kill or be injured). Some materials themselves are vulnerable to alcohol. Alternatively, in the case of surface treatment of a medical device coated with a drug such as a drug-releasing stent, damage may be caused by peeling off a polymer layer carrying the drug with alcohol.

  The present invention solves the above-mentioned conventional problems, is water-soluble (hydrophilic) at a temperature lower than a predetermined temperature (T) lower than the living body temperature, and water-insoluble (at a temperature higher than the predetermined temperature ( It is an object of the present invention to provide a phosphorcholine group-containing antithrombotic coating agent and a medical device coated with the antithrombotic coating agent, which have a temperature responsiveness that is hydrophobic.

  The antithrombotic coating agent of the present invention is an antithrombotic coating agent comprising a polymer material having a phosphorcholine group-containing polymer chain and a polymer chain of N-isopropylacrylamide or a derivative thereof. It is characterized in that it is hydrophilic at a temperature lower than the predetermined temperature (T), and is hydrophobic at a temperature higher than the predetermined temperature (T).

  The predetermined temperature (T) is preferably a temperature between 25 and 35 ° C.

  The polymer material preferably has a plurality of branched chains made of a block copolymer of a phosphorcholine group-containing monomer and N-isopropylacrylamide or a derivative thereof.

  As the phosphoroline group-containing monomer, a phosphorocholine group-containing (meth) acrylic acid ester is preferable, and 2- (meth) acryloyloxyethyl-2 '-(trimethylammonio) ethyl phosphate is particularly preferable.

  The polymerization degree of the phosphorcholine group-containing monomer is 10 to 6000, and the number average molecular weight of the polymer chain of N-isopropylacrylamide or its derivative is preferably about 3,000 to 150,000.

  The polymer material uses an iniferter as a compound having three or more N, N-dialkyl-dithiocarbamylmethyl molecular groups in the same molecule, and is irradiated with a phosphoroline group-containing monomer and N-isopropylacrylamide or a derivative thereof. A polymerized branched polymer is preferred.

  The compound having three or more N, N-dialkyl-dithiocarbamylmethyl groups in the same molecule has a benzene ring as a nucleus and three or more N, N-dialkyl-dithio as a branched chain in this nucleus. Those having a carbamylmethyl molecular group bonded thereto are preferred. For this iniferter, either a phosphorcholine group-containing monomer and N-isopropylacrylamide or a derivative thereof may be polymerized first.

  The medical device of the present invention is coated with the antithrombotic coating agent of the present invention.

  The antithrombotic coating agent of the present invention is hydrophilic and water-soluble at a temperature lower than the predetermined temperature due to the action of the polymer chain of N-isopropylacrylamide. Accordingly, this antithrombotic coating agent is dissolved in water and applied to a living body or a medical device, and then becomes hydrophobic (water insoluble) by setting it to a temperature higher than a predetermined temperature. Adhere to. The attached antithrombotic coating agent has an antithrombotic action due to the phosphorcholine group-containing polymer chain.

  This antithrombotic coating agent can be stably stored for a long time as an aqueous solution, and can also be applied to a hybrid material or a living body.

  When the polymer material constituting the antithrombotic coating agent has a plurality of branched chains composed of a block copolymer of a phosphoroline group-containing monomer and N-isopropylacrylamide or a derivative thereof, the antithrombotic coating agent is attached to a medical device or the like. The wearing power is high. That is, in the antithrombotic coating agent having a plurality of branched chains, if all of the plurality of branched chains in one molecule that are hydrophobically bonded to the surface of a medical device or the like are not peeled off simultaneously, the molecule (polymer) is removed. If the remaining branched chain is bonded even if one is peeled off, the branched chain once peeled can be rebound to the material surface. On the other hand, in the case of a linear block polymer having only one polymer chain in one molecule, if this one polymer chain is peeled off, the molecule is peeled off as it is, so that the adhesion is weak. Conceivable.

  The antithrombotic coating agent of the present invention has a phosphorcholine group-containing polymer chain and a polymer chain of N-isopropylacrylamide or a derivative thereof.

  As the above polymer material, a compound having three or more N, N-dialkyl-dithiocarbamylmethyl molecular groups in the same molecule is used as an iniferter, and a phosphoroline group-containing monomer and N-isopropylacrylamide or a derivative thereof are used as light. A branched polymer subjected to irradiation living polymerization is preferred.

  In this specification, the iniferter is a polymerization initiator that generates radicals upon irradiation with light, a function as a chain transfer agent, a function that bonds with the growth terminal to stop the growth, and a polymerization when light irradiation stops. It is a molecule that functions as a polymerization initiator / termination agent for stopping.

  As a compound having three or more N, N-dialkyl-dithiocarbamylmethyl molecular groups in the same molecule, three or more N, N-dialkyl-dithiocarbamylmethyl molecular groups are bonded as a branched chain to the benzene ring. These are preferred, and specific examples are as follows. That is, as a 3-branched chain, 1,3,5-tri (bromomethyl) benzene and sodium N, N-dithiocarbamate (sodium N, N-dithiocarbamate) can be obtained by addition reaction in ethanol. , 3,5-tri (N, N-dithiocarbamylmethyl) benzene, and four branched chains include 1,2,4,5-tetrakis (bromomethyl) benzene and sodium N, N-dithiocarbamylate ( 1,2,4,5-tetrakis (N, N-dithiocarbamylmethyl) benzene obtained by addition reaction with sodium N, N-dithiocarbamate) in ethanol. Hexakis (N, N-dithio) obtained by addition reaction of bromomethyl) benzene and sodium N, N-dithiocarbamate in ethanol Rubamirumechiru) is a benzene.

  The above iniferter is almost insoluble in polar solvents such as alcohol, but is easily soluble in nonpolar solvents. The nonpolar solvent is preferably a hydrocarbon, an alkyl halide or an alkylene halide, and particularly preferably benzene, toluene, chloroform or methylene chloride, particularly toluene and chloroform.

Monomers to be polymerized to this iniferter are phosphoroline group-containing monomers and N-isopropylacrylamide, and any of them may be polymerized first. In addition, (meth) acrylate represents an acrylate and / or a methacrylate.
As a phosphoroline group-containing monomer, the following general formula (1)

(Wherein R ¹ and R ² represent a hydrogen atom or a methyl group, and n represents a number of 1 to 10), and a phosphorocholine group-containing (meth) acrylic acid ester is preferred. 2- (meth) acryloyloxyethyl-2 ′-(trimethylammonio) ethyl phosphate, 2- (meth) acryloyloxypropyl-2 ′-(trimethylammonio) ethyl phosphate, 2- (meth) acryloyloxyethoxyethyl- 2 '-(trimethylammonio) ethyl phosphate, 2- (meth) acryloyloxydiethoxyethyl-2'-(trimethylammonio) ethyl phosphate, 2- (meth) acryloyloxytriethoxyethyl-2 '-(trimethylammoni E) Ethyl phosphate is preferred. Among these, 2- (meth) acryloyloxyethyl-2 ′-(trimethylammonio) ethyl phosphate is particularly preferable from the viewpoints of economy and availability.

  To react the iniferter with the above monomer, prepare a raw material solution containing the iniferter and one monomer, and irradiate it with light, whereby the first step reaction product in which the one monomer is bonded to the iniferter Then, the other monomer is block copolymerized with each branched chain of the first step reaction product. As the solvent in this case, the solvent of the above iniferter may be used.

  The concentration of the phosphorcholine group-containing monomer in the raw material solution is preferably 0.1 M or more, for example, 0.1 to 2.5 M. The concentration of N-isopropylacrylamide or a derivative thereof is preferably 0.1M or more, for example, about 0.1 to 2.5M.

  The concentration of the iniferter is preferably about 0.1 to 100 mM.

200-400 nm is suitable for the wavelength of the light to irradiate. The irradiation time of the light depends on the irradiation intensity, about 1 to 60 minutes are preferred, about 1 to 30 minutes at a low irradiation intensity of about 1μW ^/ cm ² ~10mW ^/ cm 2 is particularly preferred.

  The reaction product produced in the light irradiation step (first light irradiation step) using one monomer is preferably purified, and then preferably dissolved in alcohol and the second light irradiation using the other monomer. Perform the process. As the alcohol, methanol or ethanol, particularly methanol is preferable. The concentration of the first step reaction product in the alcohol solution is preferably about 0.01 mM to 100 mM.

  The polymerization degree of the phosphorcholine group-containing monomer is particularly preferably 10 to 6000, and the number average molecular weight of the polymer block of N-isopropylacrylamide or a derivative thereof is particularly preferably about 3,000 to 150,000.

  Since the target branched polymer is produced in the reaction solution by this light irradiation, the antithrombotic coating agent comprising the branched polymer can be obtained by purification if necessary.

  The molecular weight of this branched polymer depends on the number of branched chains, but is preferably about 3,000 to 600,000, particularly 3,000 to 150,000, and more preferably about 3,000 to 100,000.

  The branched polymer thus produced has a polymer block of N-isopropylacrylamide or a derivative thereof. This polymer chain of N-isopropylacrylamide or a derivative thereof has a temperature dependency that becomes hydrophilic at a temperature lower than 30 ° C. and becomes hydrophobic at a temperature higher than 30 ° C. It is insoluble in water at temperatures higher than 0 ° C. and becomes water-soluble at temperatures lower than about 30 ° C.

  Accordingly, an antithrombotic coating agent aqueous solution (concentration is preferably about 3 to 150 mg / mL) at a temperature lower than about 30 ° C., for example, about 10 to 25 ° C. is attached to a living body or a medical device by application or the like. The antithrombotic coating having a water-insoluble, phosphoroline group-containing polymer block is formed by raising the temperature to a higher temperature and drying as necessary. In the case of a living body, the temperature increase at this time is performed by the body temperature of the living body. The formed antithrombotic coating has a plurality of branched chains containing a polymer block of N-isopropylacrylamide or a derivative thereof, and thus has high adhesion to living bodies or medical devices and is difficult to peel off.

This medical device is used for parts that come into contact with blood, such as an artificial heart, an artificial heart valve, an artificial blood vessel, a blood vessel catheter, a blood vessel stent, a cannula, an artificial heart lung, a blood vessel bypass tube, an aortic balloon pumping, a blood transfusion device, and an extracorporeal circuit. Examples thereof include medical devices to be used. When applied to a medical device, it is preferable to attach about 0.1 to 30 mg of the antithrombotic coating agent per 1 cm ² of the surface of the medical device.

Example 1
i) Synthesis of iniferter According to the following reaction formula, 1,2,4,5-tetrakis (NN-diethyldithiocarbamylmethyl) benzene was synthesized as follows.

  1.0 g of 1,2,4,5-tetrakis (bromomethylbenzene) and 4.0 g of sodium N, N-diethyldithiocarbamate were added to 100 mL of ethanol and stirred at room temperature for 4 days under light shielding. The precipitate was filtered, dried under reduced pressure, dissolved in 200 mL of chloroform, 150 mL of water was added, liquid-liquid extraction separation was performed, and sodium bromide was removed. After repeating this operation three times, the chloroform layer was dried over magnesium sulfate for 24 hours, filtered, added with n-hexane, recrystallized and purified, and white 1,2,4,5-tetrasakis ( Acicular crystals of (N, N-diethyldithiocarbamylmethyl) benzene were obtained (yield 90%). The starting material peak disappeared by high performance liquid chromatography, and it was confirmed that the purified product was a single substance.

The measurement result of ¹ H NMR (in CDCl ₃ ) is as follows: σ1.26-1.31 ppm (t, 24H, CH ₂ CH ₃ ), σ 3.69-3.77 ppm (q, 8H, N (CH ₂ CH ₃ ) ₂ ), Σ3.99-4.07 ppm (q, 8H, N (CH ₂ CH ₃ ) ₂ ), σ 4.57 ppm (s, 8H, Ar—CH ₂ ), σ 7.49 ppm (s, 2H, Ar—H) It became.

ii) Synthesis of cationic homopolymer comprising 4-branched star polymer by photopolymerization According to the following reaction formula, 1,2,4,5-tetrakis [(N, N-diethyldithiocarbamyl) -A homopolymer composed of poly (methacryloyloxyethyl-2 '-(trimethylammonio) ethyl phosphate) -methyl] benzene (hereinafter sometimes referred to as pMPC) was synthesized.

That is, 45.6 mg of 1,2,4,5-tetrakis (N, N-diethyldithiocarbamylmethyl) benzene synthesized by the above i) was dissolved in 20 mL of chloroform, and methacryloyloxyethyl-2 ′-(trimethylammonium) was dissolved. E) 10 g of ethyl phosphate was added and mixed, and the whole amount was adjusted to 50 mL with chloroform. After purging with high-purity nitrogen gas for 5 minutes with vigorous stirring in a quartz cell, ultraviolet light was irradiated for 20 minutes with a 200 W high-pressure mercury lamp. The irradiation intensity was adjusted to 1 mW / cm ² (250 nm) using an illuminometer (UVR-1, TOPCON, Tokyo, Japan). After 20 minutes, the polymerization solution was concentrated with an evaporator, purified by reprecipitation of the polymer with diethyl ether, dissolved in a small amount of water, filtered through a 0.2 μm filter, and freeze-dried to form a 4-branch star type. A homopolymer pMPC was obtained (polymerization rate 40%). The molecular weight was measured as 30,000 by GPC.

iii) Synthesis of an antithrombotic coating agent comprising a polymer material (4-branched pMPC-b-pNIPAM) by block copolymerization of N-isopropylacrylamide with a homopolymer In accordance with the following reaction formula, tetrakis [(N , N-diethyldithiocarbamyl-poly (N-isopropylacrylamide) -block- (2-methacryloyloxyethyl-2 ′-(trimethylammonio) ethyl phosphate) -methyl] benzene (hereinafter referred to as pMPC-b-pNIPAM) In some cases).

  That is, the whole amount of the 4-branched pMPC homopolymer synthesized in the above ii) was transferred to a 1 liter flask, re-precipitated by adding about 800 mL of diethyl ether, and diethyl ether was removed by decantation. About 50 mL of toluene was added thereto to dissolve the polymer component, and diethyl ether was added again to reprecipitate the polymer component. This operation was repeated three times. The polymer component was dissolved in about 20 mL of methanol, and 0.5 g of N-isopropylacrylamide (NIPAM) was mixed to adjust the total amount to 60 mL with methanol. Photoirradiation polymerization was performed under the same conditions as in ii) to obtain a polymer material composed of a block polymer (4-branched pMPC-b-pNIPAM) of 4-branched pMEA and poly N-isopropylacrylamide (pNIPAM). The molecular weight was determined to be 46,000 by GPC.

v) Coating treatment 20 mg of the above polymer material (4-branched pMPC-b-pNIPAM) was dissolved in water to adjust the total amount to 2 mL. 10 μL of this solution was homogeneously cast onto a 2 cm × 3 cm square PET film and dried by a dryer to carry out coating treatment. The film was placed on a plate temperature-controlled at 37 ° C., and the contact angle was measured. The contact angle in this state was 15 °, and the hydrophilization phenomenon of the film surface presumed to be caused by pNIPAM was confirmed. When this film was continuously washed with warm water at 37 ° C., the contact angle of the surface gradually increased with the washing time and reached a plateau at about 44 °. Although the contact angle of the PET film approached 74 °, it was confirmed that the polymer material was insolubilized and fixed.

vi) Confirmation of antithrombogenicity Human peripheral blood was collected, and immediately applied onto a PET film as it was, and then rinsed lightly with physiological saline after a while to observe the adhesion of the thrombus on the surface. In the PET film, thrombus generation was confirmed in about 5 minutes, but in the coated PET film, thrombus generation was not confirmed even after 20 minutes.

Comparative Example 1
(N, N-diethyldithiocarbamylmethyl) benzene was prepared in the same manner as i) of Example 1 except that (bromomethyl) benzene was used instead of 1,2,4,5-tetrakis (bromomethylbenzene). Synthesized. The purified product was a colorless liquid, and the yield was about 93%.

The measurement results of ¹ H NMR (in CDCl ₃ ) are σ7.41 to 7.27 ppm (m, 5H, Ar—H), σ4.54 ppm (s, 2H, Ar—CH ₂ —S—), σ4.08 to 4.01 ppm (q, 2H, N—CH ₂ —), σ 3.72 ppm (q, 2H, N—CH ₂ —), σ 1.25-1.31 ppm (m, 6H, —CH ₂ —CH ₃ ) became.

  Next, a linear block polymer pMPC-b-pNIPAM having only one linear chain was synthesized according to the above methods ii) and iii), and the coating treatment and water washing on the PET film of (v) above The experiment was conducted. The contact angle increased from 15 ° to 74 ° just by washing lightly at 37 ° C., and it was confirmed that the coated polymer was quickly peeled off from the PET film.

  From the above, the superiority of the branched structure of Example 1 was confirmed. That is, in the antithrombotic coating agent of Example 1, the molecule is peeled from the material surface unless all of the pNIPAM blocks of a plurality of branched chains in one molecule that are hydrophobically bonded to the material surface are peeled from the material surface at the same time. Even if one peels, if the remaining branched chain is bonded, the branched chain once peeled can be recombined, so it is considered that it is firmly fixed on the material surface. On the other hand, the linear block polymer synthesized in Comparative Example 1 has only one p-NIPAM polymer chain in one molecule. If this P-NIPAM polymer chain peels from the material surface, the molecule remains as it is. Since it peels from the surface of the material, it is considered weak against the cleaning treatment.

vi) Confirmation of Surface Immobilization 0.1 μL of the solution of the block polymer (concentration 20 mg / 2 mL) of Example 1 and Comparative Example 1 was applied and dried on the gold-deposited surface of the crystal oscillator, and the frequency of QCM was measured. I investigated. Thereafter, it was immersed in water at 37 ° C. for 10 minutes, dried, and then subjected to QCM measurement to examine the adsorption amount. Furthermore, it was immersed in 10 degreeC water for 10 minutes, and the desorption amount was investigated by QCM measurement. As the QCM apparatus, AFFINIX Q manufactured by Initiative was used. The results are shown in Table 1.

  As shown in Table 1, since the change at 30 Hz corresponds to 1 ng, the linear block polymer of Comparative Example 1 has a large amount of desorption even after washing at 37 ° C., confirming the three-dimensional structural significance of the branched structure of Example 1. It was done.

Claims (11)

An antithrombotic coating agent comprising a polymer material having a phosphorcholine group-containing polymer chain and a polymer chain of N-isopropylacrylamide or a derivative thereof,
The polymer material is hydrophilic at a temperature lower than a predetermined temperature (T) lower than a living body temperature, and is hydrophobic at a temperature higher than the predetermined temperature (T). .   The antithrombotic coating agent according to claim 1, wherein the predetermined temperature (T) is a temperature between 25 and 35 ° C.   3. The antithrombotic coating agent according to claim 1, wherein the polymer material has a plurality of branched chains made of a block copolymer of a phosphorcholine group-containing monomer and N-isopropylacrylamide or a derivative thereof.   4. The antithrombotic coating agent according to claim 3, wherein the phosphoroline group-containing monomer is a phosphorocholine group-containing (meth) acrylic acid ester.   5. The antithrombotic coating agent according to claim 4, wherein the phosphoric choline group-containing (meth) acrylic acid ester is 2- (meth) acryloyloxyethyl-2 '-(trimethylammonio) ethyl phosphate.   6. The degree of polymerization of the phosphorcholine group-containing monomer is 10 to 6000 in any one of claims 3 to 5, and the number average molecular weight of the polymer block of N-isopropylacrylamide or a derivative thereof is 3,000 to 150,000. An antithrombotic coating agent characterized by that.   The polymer material according to any one of claims 3 to 6, wherein the polymer material is an iniferter having a compound having three or more N, N-dialkyl-dithiocarbamylmethyl groups in the same molecule, and a phosphoroline group-containing monomer and An antithrombotic coating agent, which is a branched polymer obtained by light-irradiating living polymerization of N-isopropylacrylamide or a derivative thereof.   The compound having three or more N, N-dialkyl-dithiocarbamylmethyl molecular groups in the same molecule as defined in claim 7 having a benzene ring as a nucleus and three or more N, N- An antithrombogenic coating agent, wherein a dialkyl-dithiocarbamylmethyl molecular group is bonded.   9. The antithrombotic coating agent according to claim 7, wherein the phosphorocholine group-containing monomer is first polymerized and then N-isopropylacrylamide or a derivative thereof is polymerized.   9. The antithrombotic coating agent according to claim 7, wherein N-isopropylacrylamide or a derivative thereof is first polymerized and then a phosphoroline group-containing monomer is polymerized.   A medical device coated with the antithrombotic coating agent according to any one of claims 1 to 10.