JPS62204761A - Base material for extracorporeal recirculation therapy - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of the Invention The present invention relates to substrates for therapeutic materials used under conditions of contact with whole blood or body fluid cellular components.

For more details, please refer to 1. Materials for extracorporeal circulation treatment such as artificial livers, adsorbents for plasma components such as immunoadsorbers, cell separation and removal materials such as blood cell separators and blood cell removers, and materials for stimulating and activating immune cells. The present invention relates to a base material.

(Prior Art) As is well known, as an extracorporeal circulation treatment material used in the presence of blood cells, that is, a material for whole blood perfusion, activated carbon and 2-hydroxyethyl methacrylate (HEMA) are used.
) are in practical use, such as those coated with.

However, even with the materials currently in practical use, thrombotic substances such as platelets often adhere to and become activated by extracorporeal circulation treatment materials. As a result, biologically,
As the number of thrombotic substances such as platelets decreases, problems such as various symptoms arise, and on the surface of the treatment device, the primary function of the treatment device, for example, removal of toxic substances, is reduced due to the adhesion of thrombotic substances. In addition, the pore diameter of activated carbon is approximately several tens of amps.

It is extremely difficult to create pores with a pore diameter of several xooA to several thousand A. Therefore, it is used as an extracorporeal circulation treatment material for the adsorption and removal of substances with a molecular weight of about several thousand or less, but it is used for extracorporeal circulation treatment for proteins with a molecular weight of at least several thousand or more, which is the object of the present invention. It cannot be used as a material.

On the other hand, as a similar technology, synthetic polymers for biomaterials are disclosed in Japanese Patent Application Laid-Open No. 60-119955 and Japanese Patent Application Laid-open No. 60-119956.
No. 60-119957. They coated glass particles with a size of 250 to 350 microns with a polymer having a nitrogen-containing basic functional group, and evaluated using fresh rat blood containing heparin, and found that the blood did not clog.

However, glass particles cause glass elution during heat sterilization, which causes a serious problem of blood coagulation during extracorporeal circulation. Furthermore, in order to clearly demonstrate its suitability for the human body, the present inventors evaluated a material in which glass particles of 250 to 350 μm are coated with a polymer having a nitrogen-containing basic functional group using human whole blood containing heparin. As a result, it did not show practical platelet permeability.

Therefore, adaptation to the human body poses major problems in terms of safety and platelet adhesion and activation, and it has not yet been put to practical use.

(Problems to be Solved by the Invention) The objects of the present invention are to provide an artificial liver, an adsorbent for plasma components such as an immunoadsorber, a blood cell separator, and a blood cell separator to be used under conditions in which whole blood or body fluid cell components come into contact. Cell separation using eliminators, etc.
The object of the present invention is to provide a base material with good blood compatibility for extracorporeal circulation treatment materials such as removal materials or immune cell stimulation and activation materials.

In view of the current state of extracorporeal circulation treatment materials and synthetic polymers for biomaterials, the present inventors have determined that a suitable base material for extracorporeal circulation treatment materials has pores with a pore size of several tens of amps to several xoooox. It is a base material that can be used to produce blood cells, and there are no safety issues such as oxidation or elution. As a result, the contact angle was surprisingly 20~85.
The inventors have discovered that a substrate for extracorporeal circulation treatment, in which a water-insoluble material in the range of about 100% is coated with a polymer having a nitrogen-containing basic functional group, satisfies the object of the present invention, and has completed the present invention.

(Means for Solving the Problems) The present invention is characterized in that the surface of a water-insoluble material having a contact angle in the range of 20 to 85 degrees is coated with a polymer having a nitrogen-containing basic functional group. The present invention relates to a substrate for extracorporeal circulation treatment.

The present inventors have developed artificial livers, adsorbents for plasma components such as immunoadsorbers, and cell separation and removal materials such as blood cell separators and blood cell removers that are used under conditions where they come into contact with whole blood or body fluid cell components. , or extracorporeal circulation treatment materials such as immune cell stimulation and activation materials.

In order to obtain a base material with good blood compatibility, we extensively changed the water-insoluble material and coated the surface with a polymer to create a base material for extracorporeal circulation therapy, and to evaluate its suitability as a base material for extracorporeal circulation therapy. In order to clarify this further, we evaluated the permeability of platelets using human whole blood using heparin, which is widely used in extracorporeal circulation therapy, as an anticoagulant. The result was surprising.

The extracorporeal circulation treatment base material, which is made by coating the surface of a water-insoluble material with a contact angle in the range of 20 to 85 degrees with a polymer having a nitrogen-containing basic functional group, has no safety problems such as eluates. The present invention was completed based on the discovery that the platelet exhibits suitable platelet passage and almost no change in platelet morphology was observed.

These results demonstrate that a polymer containing nitrogen-containing basic functional groups can be coated on a water-insoluble material to form a base material for extracorporeal circulation therapy.
does not necessarily mean that any water-insoluble material is suitable;
The contact angle of the surface of a water-insoluble material is measured, and the contact angle is 2
This shows that a suitable substrate for extracorporeal circulation therapy can be created by coating the surface of a water-insoluble material with a temperature of 0 to 85 degrees with a polymer containing a nitrogen-containing basic functional group.
The reason for this is the compatibility, that is, the compatibility, between the polymer containing a nitrogen-containing basic functional group and the water-insoluble material. The present inventors took up the contact angle of air bubbles in water on a solid surface as a compatibility parameter.

It is known that the contact angle is approximately proportional to the solubility parameter, which is known as another compatibility parameter, and to hydrophilicity and hydrophobicity, which are used to determine the magnitude of interaction with water.

According to the invention, the contact angle of the surface of the water-insoluble material is 20
A range of 85 degrees was suitable. This means that if interpreted in terms of solubility parameters, suitable parameter values, similar to the contact angle values, indicate an intermediate range, and the solubility parameters correspond to contact angles of 20 degrees or less and 85 degrees or more. This value is expected to have a large difference from the solubility parameter value of the polymer having a nitrogen-containing basic functional group, and it is expected that the water-insoluble material and the polymer having a nitrogen-containing basic functional group will undergo a phase separation phenomenon. Therefore, the structure of the coating layer may be formed in an incomplete form, the thickness of the coating layer may be uneven, or the coated polymer may peel off.

Also, if interpreted from the viewpoint of hydrophilicity and hydrophobicity.

Materials with a contact angle of 20 degrees or less are more hydrophilic materials,
A material with a contact angle of 85 degrees or more is a more hydrophobic material.

Compatibility between hydrophilic and hydrophobic materials is poor, and only a limited number of polymers with nitrogen-containing basic functional groups are suitable for coating water-insoluble materials with contact angles of 20 degrees or less and 85 degrees or more. Even if the product is suitable, the coating operation must be repeated many times or heat treatment and fixation must be performed, resulting in poor reproducibility (unusable).

In addition, more hydrophilic water-insoluble materials with a contact angle of 20 degrees or less tend to form a film of water on their surfaces and have poor compatibility with polymers having nitrogen-containing basic functional groups. In addition, more hydrophilic water-insoluble materials with a contact angle of 20 degrees or less, or with a contact angle of 85
Due to its stronger hydrophilic and hydrophobic properties, the orientation of the nitrogen-containing basic functional groups of the coated polymer with nitrogen-containing basic functional groups is more uneven and uneven. It is possible that it will become.

As mentioned above, by using a water-insoluble material with a contact angle of 20 to 85 degrees, the compatibility between the coating polymer having a nitrogen-containing basic functional group and the coated water-insoluble material can be improved. The coating layer has a more uniform thickness, the structure of the coating layer and the orientation of the nitrogen-containing basic functional groups are properly formed, As a result, it is thought that a suitable substrate for extracorporeal circulation treatment with less platelet adhesion was created.

The contact angle in the present invention is the contact angle of air bubbles on a solid surface in water, W, C, Hamilton
J.

Co11oid Interface 5et-*4
0. ．． 219-222 (1972) [Double-C.
Hamilton, Journal of Colloid Interface Science, 40°219-222 (1972)
)), J-D, Andrade, J, Polym.

Sci, Polym-8ymp, 66.313-33
6 (1979) [J.D. Andrade, Journal of Polymer φ Science Polymer Symposium, 66.313-336 (1979)]. Conventionally, the contact angle measurement method of a droplet on a solid surface in air changes with the passage of time for hydrophilic materials, making it difficult to use as a physical property value of the material. In addition, as samples, molded products such as sheets and films were produced, and the contact angle was measured at least 10 times at a temperature of 25° C., and the average value was taken as the value of the contact angle of the material.

The contact angle of the water-insoluble material of the present invention is preferably in the range of 20 to 85 degrees, more preferably 30 to 80 degrees, and more preferably 40 to 85 degrees.
~75 degrees is more preferred.

In the present invention, a water-insoluble material with a contact angle in the range of 20 to 85 degrees is a solid in an aqueous liquid, and if the contact angle measured by the method described above is 20 to 85 degrees, it is an inorganic material. Although all types of compounds and organic polymer compounds are included, organic polymer compounds are preferred because they allow easier and more precise control of eluates and pores. Examples of such are polypropylene, polystyrene, polymethacrylate esters, polyacrylate esters, polyacrylic acid.

Examples include polymers and copolymers of vinyl compounds such as polyvinyl alcohol, polyamide compounds such as nylon 6.66, and polyester compounds such as polyethylene terephthalate.

In the present invention, the contact angle may be in the range of 20 to 85 degrees, and is not limited to the above examples (S.

Further, based on the similarity of chemical structure with the polymer having a nitrogen-containing basic functional group to be coated (based on compatibility), polymers and copolymers of vinyl compounds are more preferable. Styrene, p-methylstine, p
- Polymers of styrene compounds such as ethylstyrene, polymers of (meth)acrylic acid ester compounds such as methyl (meth)acrylate, ethyl (meth)acrylate, and propyl (meth)acrylate, and the above compounds and divinylbenzene, Examples include copolymers with vinyl compounds such as methacrylonitrile, vinylpyrrolidone, ethylene glycol, and methyl acrylate.

Among these, methyl (meth)acrylate, ethyl (meth)acrylate, propyl (meth)acrylate, etc.
Polymers of meth)acrylic acid ester compounds and copolymers with the above-mentioned vinyl compounds are more preferably used.

Furthermore, the water-insoluble material of the present invention is more preferably a polymer of a (meth)acrylic acid ester compound and a copolymer b'- with the above-mentioned vinyl compound, and a material containing 70 times i4 or more of methyl methacrylate. Give results.

The shape of the water-insoluble material may be spherical, granular, membrane-like, hollow fiber-like, filament-like, etc., but particle-like shapes such as spherical, single-grain, etc. This is because, when filled with a water-insoluble material, the area that comes into contact with blood becomes larger, increasing the efficiency of removing toxic substances, etc.

Particulate form is preferred.

Furthermore, when the water-insoluble material is in the form of particles, those having a particle size of 50 to 2000 microns give more preferable results as the water-insoluble material of the present invention. Particle size is 50μ
If it is smaller than 2,000 μm, the distance between the particles becomes small, and platelets etc. will adhere to it, making it easier for them to get clogged. If it is larger than 2000 μm, it will be difficult for platelets etc. to adhere to it, but the area that comes into contact with blood will be small, making it difficult for toxic substances etc. Since the removal efficiency decreases, it is not preferable as a substrate for extracorporeal circulation treatment.

Further, a material having a particle size of 100 to 1000 μ gives more preferable results as the water-insoluble material of the present invention.

The "nitrogen-containing basic functional group" used in the present invention is a functional group that has a positive charge on a nitrogen atom and can become a cation in an acidic aqueous solution. Examples of such functional groups include primary amine groups, secondary amino groups, tertiary amino groups, quaternary ammonium groups, and nitrogen-containing aromatic ring groups such as pyridyl groups and imidazolinyl groups. Therefore, examples of the polymer having a nitrogen-containing basic functional group used in the present invention include vinylamine: 2-vinylpyridine, 4'-vinylpyridine, 2-methyl-5-vinylpyridine, 4-vinylimidazole, N -vinyl-2-ethylimidazole,
Vinyl derivatives of nitrogen-containing aromatic ring compounds such as N-vinyl-2-methylimidazole; dimethylamine ethyl (meth)
Acrylate, diethylaminoethyl (meth)acrylate, dimethylaminoethyl (meth)acrylate, 3
- Acrylic acid and methacrylic acid derivatives such as dimethylamine-2-hydroxypropyl (meth)acrylate; Acrylic acid amides such as N-dimethylaminoethyl (meth)acrylic acid amide, N-diethylaminoethyl (meth)acrylic acid amide; Examples include polymers containing methacrylic acid amide derivatives; styrene derivatives such as p-dimethylaminomethylstyrene and p-diethylaminoethylstyrene; and derivatives made into quaternary ammonium salts with alkyl halides, etc. of the above-mentioned vinyl compounds. It will be done.

Among these, dimethylaminoethyl (meth)acrylate, diethylaminoethyl (meth)acrylate, p-dimethylaminomethylstyrene, p--
/ethylaminoethylstyrene and the like.

This means that by making the water-insoluble material a water-insoluble material with a contact angle of 20 to 85 degrees, the compatibility with the above-mentioned polymer becomes favorable, and therefore problems such as polymer peeling do not occur, and weak cation It is assumed that the material has a uniform surface structure and a uniform coating layer thickness, and as a result, it has good permeability for platelets in an evaluation using human blood, making it a suitable substrate for extracorporeal circulation treatment. It is thought that

The polymer having a nitrogen-containing basic functional group in the present invention is preferably a polymer whose surface has a contact angle of 10 to 95 degrees. As mentioned in 5 above, a polymer having a nitrogen-containing basic functional group also has a nitrogen-containing basic functional group with a contact angle of 10 to 95 degrees by measuring the contact angle of its surface in the same way as a water-insoluble material. By coating a water-insoluble material with a contact angle of 20 to 85 degrees with a polymer, a more preferable substrate for extracorporeal circulation treatment can be obtained. This is due to the compatibility between the coating polymer having nitrogen-containing basic functional groups and the coated water-insoluble material. If interpreted in terms of solubility parameters, the solubility parameter values of both are similar. This prevents the occurrence of phase separation at the interface, prevents peeling of the coated polymer having nitrogen-containing basic functional groups, and allows the structure of the coating layer to be formed appropriately, resulting in the adhesion of platelets. It is thought that a suitable base material for extracorporeal circulation treatment with less oxidation can be created.

Furthermore, a contact angle of the polymer having a nitrogen-containing basic functional group of the present invention of 15 to 85 degrees gives more preferable results.

In addition, the polymer having a nitrogen-containing basic functional group referred to in the present invention is preferably a copolymer of a vinyl compound and a monomer having a nitrogen-containing basic functional group, and the nitrogen content in the functional group is
The weight range is preferably 0.05 to 3.5. Furthermore, more preferable results are obtained when the nitrogen content is in the range of O,1 to 2.5 by weight. The nitrogen content referred to herein is the weight range of nitrogen atoms in the above functional group in the total polymer.

By coating a water-insoluble material having a contact angle of 20 to 85 degrees with a polymer having a nitrogen-containing basic functional group within the above range, a preferred substrate for extracorporeal circulation treatment can be obtained. This means that by making a copolymer with a defined nitrogen content, a surface structure with controlled cationic properties can be obtained, and since it is a copolymer with a vinyl compound, it is easier to control the contact angle (therefore, This is thought to be because the compatibility with water-insoluble materials having a contact angle of 20 to 85 degrees is better, resulting in a base material for extracorporeal circulation treatment with a surface structure with better cationic control.

The vinyl compounds mentioned here include alkyl (meth)acrylates such as 2-hydroxyethyl methacrylate, methyl (meth)acrylate, ethyl (meth)acrylate, and n-butyl (meth)acrylate;
Examples include amides such as acrylamide and N-methyl (meth)acrylamide, N-vinylpyrrolidone, vinyl acetate, and styrene.

Furthermore, the vinyl compound of the copolymer of the vinyl compound shown above and the monomer having a nitrogen-containing basic functional group is %2-
More preferred is hydroxyethyl methacrylate. This means that by using 2-hydroxyethyl methacrylate as the vinyl compound, it becomes a copolymer with a micro-heterogeneous phase separation structure having a hydrophilic part and a hydrophobic part, but the contact angle is 20 to 85. By coating a water-insoluble material with a high degree of compatibility, problems such as peeling of the copolymer do not occur, and the thickness and structure of the coating layer are uniform. It is thought that a non-uniform structure was formed, and as a result, a suitable base material for extracorporeal circulation treatment with good platelet permeability was created.

The monomer having a nitrogen-containing basic functional group mentioned here includes the monomer of the polymer having a nitrogen-containing basic functional group shown above. The monomer having a nitrogen-containing basic functional group is
More preferred are dimethylamine ethyl (meth)acrylate, diethylaminoethyl (meth)acrylate, p-diethylaminomethylstyrene, and p-diethylaminoethylstyrene.

Furthermore, copolymers of vinyl compounds and monomers having nitrogen-containing basic functional groups include block copolymers, graft copolymers, random copolymers, etc. Preferably, the polymer has a microdomain structure with an average length of 100A to 100μ.

The copolymer of the vinyl compound and the monomer having a nitrogen-containing basic functional group is most preferably a random copolymer.

Regarding this effect, contact angle of water-insoluble material is 20 to 8.
Because it is a water-insoluble material of 5 degrees, it has good compatibility with random copolymers.

A vinyl compound and a monomer having a nitrogen-containing basic functional group copolymerize in a more random manner, resulting in a form in which amino groups are present in a more random manner on the surface that comes into contact with blood, which is a preferred method with less interaction with platelets. It is thought to be used as a base material for extracorporeal circulation therapy.

In addition, the water-insoluble material of the present invention coated with a polymer having a nitrogen-containing basic functional group was tested in accordance with the Ministry of Health and Welfare's quality standards for adsorption-type blood purifiers, and an eluate test and an outflow foreign matter count test were conducted. , all were within the standards. Furthermore, in the blood coagulation ability test using the Lee-White (9-White) method, all of the base materials obtained by coating the water-insoluble material of the present invention with a polymer having a nitrogen-containing basic functional group contained glass. The blood coagulation time was observed to be longer than the substrate coated with the polymer having nitrogen basic functional groups.

In addition, the average pore diameter of the base material obtained by coating the water-insoluble material of the present invention with a polymer having a nitrogen-containing basic functional group was measured by mercury intrusion method (for example, Catalyst Engineering Course 4. Basic Measurement Methods of Catalysts, edited by Catalysis Society of Japan). , Chie Shokan.

(pages 69 to 73), the number 1 OA
We were able to produce rice plants of up to several hundred OA.

These results indicate that the base material obtained by coating the water-insoluble material of the present invention with a polymer having a nitrogen-containing basic functional group is
It is a base material that can make wires with different diameters from 0A to several thousand amps, and it is also a monopolymer (there are no safety problems such as release and elution of impurities, etc., and it is resistant to blood oranges such as platelets). This indicates that there is almost no adhesion or activation of the substance.

Below, (1) a method for producing a water-insoluble material, (2) a method for producing a polymer having a nitrogen-containing basic functional group, and (3) a coating method, conditions, etc. will be exemplified.
Of course, the invention is not limited to this example.

(1) Method for producing water-insoluble materials The method for producing the water-insoluble material of the present invention is bulk polymerization.

Generally known methods such as solution polymerization, suspension polymerization, and emulsion polymerization involve purchasing commercially available monomers and adding additives, a polymerization initiator, and a solvent for dissolving the monomers to each polymer. This is done using a manufacturing method. For example, in styrene to divinylbenzene copolymer.

By stirring in the coexistence of styrene, ethylbenzene, divinylbenzene, toluene, octatool, and AIBN (azobisisobutyronitrile), porous particles having a diameter of about 50 to 1000 μm can be produced. Particles with various particle sizes and pore sizes can also be produced by radical polymerization in a suspension polymerization system.

(2) Method for producing a polymer having a nitrogen-containing basic functional group There are no particular restrictions on producing the polymer of the present invention, and addition polymerization including radical polymerization, anionic polymerization, etc. using monomers, ring-opening polymerization, etc. , polymerization by dehydrohalogenation, condensation reaction, etc. can be used. Furthermore, methods by polymer reaction can also be adopted. For example, a predetermined raw material polymer can be aminated by a known method, or if necessary, hydroxyl groups can be introduced. When producing graft polymers, pressure can also be obtained by copolymerization of macromers and other monomers.

It is also possible to perform a graft reaction on the polymer raw material.

(3) Coating method and conditions As a coating method, all known methods such as a spray method and a dipping method can be used. Here, details of the immersion method will be explained.

A solution is prepared in advance by dissolving a polymer having a nitrogen-containing basic functional group in a polymer solvent. The polymer solvent is a solvent that uniformly dissolves the polymer and facilitates impregnation or application of the polymer onto the membrane surface.In the present invention, basically, a solvent that can dissolve the polymer is used. If so, all are available. An appropriate solvent must be selected in consideration of ease of removal, safety if a trace amount remains, etc. In the present invention, such solvents are preferably lower alcohols such as methanol and ethanol, acetone and dimethylformamide, and mixtures of K and water.

Water-insoluble particles 35- of various spherical diameters were immersed in this solution 100- for about 5 minutes at room temperature under a nitrogen atmosphere (with occasional stirring), and after removing the excess solution by suction on a glass filter, the Dry with nitrogen on a glass filter for 20 minutes while balancing the amount of nitrogen gas input and the amount of nitrogen gas suctioned. Then in a vacuum dryer at room temperature, 755 wmHf
Dry for 24 hours under the above conditions.

The method of using the extracorporeal circulation treatment substrate in which the water-insoluble material of the present invention is coated with a polymer having a nitrogen-containing basic functional group will be exemplified below, but the present invention is of course not limited to this example. .

The base material of the present invention is used for the purpose of the present invention while holding a ligand that interacts with the target substance in some way.

The ligands held by the base material of the present invention can be freely selected depending on the purpose, and some of them are exemplified below.

For the treatment of systemic lupus erythematosus, homopolymers or copolymers of mono-, di-, and trinucleotides such as adenine, guanine, cytosine, uracil, and thymine, and naturally occurring DNA are used for adsorption and removal of anti-nuclear antibodies and anti-DNA antibodies. Nucleic acids such as , RNA, as well as nucleic acid bases, sugars, oligosaccharides, and hydrophobic organic compounds such as tryptophan and phenylalanine can be used. In addition, it is used to adsorb and remove DNA, RNA, and ENA present in the blood.
Heavy chain DNA antibody, anti-double strand DNA antibody, anti-RNA antibody,
Anti-nucleic acid antibodies such as anti-ENA antibodies, basic compounds such as methylated albumin actinomycin D, etc. can be used.Furthermore, for adsorption and removal of immune complexes in blood, C1
Complement components such as q, specific proteins such as protein A, anti-hepechain constant phase II antibodies, antibodies against immune complexes such as rheumatoid factor, and hydrophobic organic compounds such as tryptophan and phenylalanine can be used.

For the treatment of chronic rheumatoid arthritis and malignant rheumatoid arthritis,
Modified r-globulin, denatured by chemical denaturation (aggregation) methods such as urea, guanidine hydrochloride, mercaptoethanol, surfactants, organic solvents, physical denaturation (aggregation) methods such as heat, ultrasound, gas bubbling, etc. Immunoglobuli?
, aggregated r-globulin, aggregated immunoglobulin, the Fe part of immunoglobulin, the second phase of the heavy chain constant part of immunoglobulin, and their modified forms according to the above-mentioned modification methods, and other antigen-like substances against rheumatoid arthritis factors.

Anti-rheumatoid factor antibodies and hydrophobic organic compounds such as tryptophan and phenylalanine can be used. Also, for removing immune complexes from rheumatism.

Complement components such as C1q, 41FM proteins such as protein A, anti-heavy chain constant region 81!2 phase antibodies, antibodies against immune complexes such as rheumatoid factor, and hydrophobic organic compounds such as tryptophan and phenylalanine can be used. .

For the treatment of Hashimoto's disease, thyroglobulin, a fractionated component of thyroid microsomes, can be used.

For the treatment of myasthenia gravis, neuromuscular acetylcholine receptor fraction components, tryptophan, adenine, etc. can be used.

Glomerular basement membrane components for the treatment of glomerulonephritis; platelet membrane components and platelet granule fraction components for the treatment of idiopathic thrombocytopenic purpura; transcortisin and anti-cortiscin antibodies for the treatment of Cushing syndrome. can be used.

For the prevention and treatment of hepatitis, hepatitis A virus, Bffl
Antibodies against surface antigens of viruses such as hepatitis viruses can be used.

Anti-angiotensin antibodies can be used to treat hypertension, and heparin, anti-boprotin antibodies, and polyanions such as polyacrylic acid, polyethylene sulfonic acid, and dextran sulfate can be used to treat hyperlipidemia.

For the treatment of immune diseases based on lymphocyte abnormalities, anti-lymphocyte antibodies such as anti-B cell antibodies, anti-pressor T antibodies, anti-lymphocyte antibodies, anti-monocyte antibodies, anti-natural killer cell antibodies, and anti-erythrocyte antibodies are used. Lectins such as or peanut lectin, histamine, etc. can be used.

Anti-erythrocyte antibodies and anti-platelet antibodies can be used for membrane diseases of red blood cells and platelets.

For cancer treatment such as breast cancer, protein A, anti-immunoglobulin antibodies, and antibodies against immunosuppressive factors can be used.

As an immune cell stimulation, activating stimulus, and activating agent,
Pressure lymphokine, cytokines, lectin, protein A, lipopolysaccharide (L) for inducing antitumor cells.
PS), 0K432, oligosaccharides, nucleobases, nucleosides, nucleotides, pyrazolo(3,4-a) pyrimidines and derivatives thereof, or organic compounds having anionic groups can be exemplified as ligands.

Cytokines include interleukin-11, which is natural or obtained using genetic engineering.
Interyukin 2. Examples include r-interferon and tumor necrosis factor (TNF).

As a lectin, Phaseolus vulgaris (Pha
Red bean lectin (PHA) from Concanavalia ensiformis (Seolus vulgaris), Concanavalia enaifor
Concanavalin A (Con A) derived from mLs)
* Rice Terrier/Aori Panda (Wisteria)
aoribanda) and Lens culinaris (Lens culinalis).
lentil lectin (LCH) derived from
), Phytolacca americana (Phyto-1ac
ca americana), glycine max (Glyci
Soybean lectin (SBA) derived from Phaseolus limensis (Phaseolus lime)
Glue mamame lectin (LBA) from Lobina pseudoacacia (R□bina pseudoaca)
cassia lectin (RPA) derived from caeia
, Sophora japonica (5opnora japonica)
dog bean lectin (SJA) derived from ca ),
Piaunlaativum
)-derived pea lectin (PSA).

Examples include Vicia faba-derived Vicia faba lectin (VFA).

The oligosaccharide is N-acetylglucosamine.

It is composed of three or more of N-acetylgalactosamine, galactose, fucose, mannose, glucose, and sialic acid. For example, sialic acid.

Galactose, N-acetylglucosamine, an oligosaccharide consisting of 7 courses, or galactose.

N-acetylglucosamine, an oligosaccharide consisting of fucose, for example.

Gal/l-+4 G1cNAcβ1-+ 3 Gal
An example is an oligosaccharide having the structure βI↑Fucα1.

Any natural or synthetic nucleobases, nucleosides, nucleotides, and chemically modified derivatives thereof can be used.

Namely, adenine, 1-methyladenine, 2-methyladenine, N-('IJ-6-ylcarbamoyl)-
L-threonine, N@(Δ2-isopentenyl)atenine, 2-methylthio-N'-(Δ2-inpentenyl)adenine, 2-hydroxyadenine, N6-methyladenine, N"I N'-dimethyladenine, N"
(eta-4-hydroxyisopentenyl)adenine, zeatin, 2-aminoadenine, quanine, l-methylguanine, N2-methylguanine, N”,N”-dimethylguanine, 7-methylguanine, cytosine, 3-methylcytosine , N4-acetylcytosine, N4-methylcytosine, 5-methylcytosine, 5-hydroxymethylcytosine, 2-thiocytosine, uracil, 3-methylcytosine, thymine, 4-thiocracil, 5-hydroxyuracil, 5-hydroxymethyluracil, 5-medoxyuracil, 5-carboxymethyluracil, 2-thiothymine, 5-carboxymethyl-2-thiouracil, 5-
(methoxycarbonylmethyl)-2-thiouracil, 5
-(N-methylaminomethyl)-2-thiouracil, 5
．． 6-cyhydrouracil, 5.6-cyhydrothymine, 5
-(butrezitzmethyl)uracil, S(t)5-(4,
Nucleic acid bases such as 5-dihydroxypentyl) uracil, orotic acid, Y, wiputin, peroxywiputin, hypoxanthine, 1-methylhyboxanthin, xanthine, uric acid, adenosine, 2-methyladenosine, guanosine, 1-methylguanosine, inosine , cytidine, uridine, and other nucleosides.

Adenosine 3-phosphate, guanosine 3-phosphate.

Examples include nucleotides such as cytidine 3-phosphate and uridine 3-phosphate.

As the organic compound having an anionic group, any compound having an anionic group can be used, but
Especially sulfonic acid groups and carboxylic acid groups.

Preferred are organic compounds having at least one of a phosphoric acid group, an arsonic acid group, a selenonic acid group, and more preferably a sulfonic acid group, a carboxylic acid group, and a phosphoric acid group.

The ligand for stimulating and activating immune cells is not limited to the above examples, and any ligand that can be immobilized on the substrate of the present invention to induce antitumor cells can be used.

Ligands that can be used in the present invention are not limited to the above examples, but include lectins such as congnitinin, concanavalin A, phytohemaagglutinin, and bourk-wietomidien, nucleic acids, amino acids, lipids, glycolipids, and protamines. , heparin.

Known substances that can interact with target substances such as antigens, antibodies, enzymes, substrates, coenzymes, and glycoproteins can be used.

Furthermore, the base material of the present invention can be used with two or more ligands retained therein. Furthermore, two or more types of materials holding ligands can be used in combination.

In the present invention, any known method can be used to immobilize the organic ligand on the surface of the substrate, such as covalent bonding, ionic bonding, physical adsorption, embedding, or precipitation insolubilization on the surface of the substrate. Considering the dissolution properties of the compound, by covalent bonding.

It is preferable to use it after being fixed or insolubilized. Therefore, it is possible to use commonly known methods of activating an immobilized enzyme, a substrate used in affinity chromatography, and a method of binding to a ligand. Further, if necessary, a molecule (spacer) of arbitrary length can be introduced between the base material and the ligand.

When bonding the hydroxyl heligant on the base material, the base material can be bonded using the cyanogen halide method, epichlorohydrin method, bisepoxide method, halogenated triazine method, bromoacetylbromide method, ethyl chloroformate method, carbonyldiimidazole method, etc. It can be activated by These functional groups can react with nucleophilic reactive groups such as amine groups, hydroxyl groups, carboxyl groups, and thiol groups of the ligand to form covalent bonds.

The material of the present invention in which a ligand is immobilized on a base material can be used by being filled and held in a container equipped with an inlet and outlet for a body fluid.

The drawing shows an example of a device using the material of the present invention in which a ligand is immobilized on a base material.A body fluid inlet 5 is connected to one end opening of a cylinder 2 through a backing 4 with a filter 3 stretched inside. A cap 6 having a body fluid is screwed into the opening at the other end of the cylinder 2, and a cap 8 having a body fluid lead-out lower part is screwed into the other end opening of the cylinder 2 through a backing 4' having a filter 3' stretched inside to form a container. A device 1 in which a material layer 9 is formed by filling and holding a material in the gaps 3 and 3'.
It is.

The material layer 9 may be filled with a single material having a substrate pressure ligand of the present invention fixed thereon, or may be mixed with or laminated with other materials. The volume of material layer 9 is:

When used for extracorporeal circulation, approximately 50-400° is appropriate.

When the device of the present invention filled with a material in which a ligand is immobilized on a base material is used for extracorporeal circulation, there are two methods as shown below by Ohji. One K is 1. Blood taken from the body is separated into two or more components using a centrifuge or a model plasma separator, and then the plasma component or blood cell component is passed through the device,
After purifying, regenerating, and activating immune cells, they are returned to the body together with plasma components or blood cell components that have not passed through the device.The other method is to directly nucleate blood taken from the body. This is a method of passing it through a device.

The method for passing body fluids may be either continuous or intermittent, depending on clinical needs or equipment conditions.

The liquids used in the present invention include whole blood, a patchy coat layer separated using a centrifuge, lymphocyte fluid,
Furthermore, all fluids containing body fluid cell components such as bone marrow fluid and plasma are included. It can also be effectively used for plasma separated using a model centrifuge or the like.

(Example) Next, the present invention will be described in more detail with reference to Examples.

Evaluation of Platelet Passability A) Preparation of Evaluation Column A base material for extracorporeal circulation treatment in which a water-insoluble material is coated with a polymer having a nitrogen-containing basic functional group.

A cylindrical column with a diameter of 5 φ and a length of 10m was placed at a vacuum level of 20
The column is filled while flowing physiological saline under suction of 0 to 300 μHf (80 mesh polyester mesh is attached to the inlet and outlet of the column). It is then kept in twilight for 16 hours.

B) Calculation of human blood permeability and platelet permeability 0, 1 d/
Heparinized food (l uni
t/d: heparin concentration) is flowed with a syringe-type micropump. Next, human peripheral blood (15 units/d: heparin concentration) was injected at room temperature (20°C) at 0.1-/rn in
flow at the speed of The time when the saline in the column is pushed out from the column outlet is set as sampling 0 time, and the sample bottle blood containing ethylenediaminetetraacetic acid=sodium is sampled from the column outlet. BRECHER-CRONKI within 2 hours after finishing sampling
The number of platelets and morphological changes at the column inlet and column outlet were measured using the TE method to determine suitability as a substrate for extracorporeal circulation therapy.

Platelet permeability of the column (1) Example 1 As water-insoluble materials, methyl methacrylate-divinylbenzene copolymer (go:zofcf#I), styrene-divinylbenzene copolymer (80:2OItfi)
%), sheets of vinyl alcohol-triallyl isocyanurate copolymer and particles of 350 to 500μ were prepared, and the contact angle of air bubbles in water was measured. -A random copolymer film of hydroxyethyl methacrylate and diethylaminoethyl methacrylate (2-hydroxyethyl methacrylate content: 90 moles) was produced, and the contact angle of air bubbles in water was measured. Next, 2 of the above random copolymer of 2-hydroxyethyl methacrylate and diethylaminoethyl methacrylate was prepared.
4. Prepare a methanol solution of vrt7'V.

The above water-insoluble material (spherical diameter 350 to 5 ooμ) was coated according to the coating method and pressure described above to prepare an evaluation column and evaluated using one person's blood.

Table 1 shows changes in platelet passage over time and average values.

Table 2 shows an outline of the materials used.

Table 1 Contact angle of 2-human cylinder xyethyl methacrylate-diethylaminoethyl methacrylate copolymer; 19±4
([) Table 2 Comparative Example 1 The water-insoluble materials were glass particles (particle size 350 to 500 μ) and polyethylene particles (particle size 350 to 500 μ),
The coating was carried out in the same manner as in Example 1, using the same polymer having a nitrogen-containing basic functional group as in Example 1.

Further, according to the method described above, an evaluation column was prepared and evaluated using one person's blood. Table 3 shows the changes in platelet permeability over time and the average value, and Table 4 shows an outline of the materials used.

Table 6 Contact angle of polymer of 2- and droxyethyl methacrylate; 17 ± 3 (2) Comparative Example 2 The same styrene-divinylbenzene copolymer as in Example 2 was used as the water-insoluble material, and the coating agent As, 2-hydroxyethyl methacrylate polymer 2
%wt/'V methanol solution and a 2'lpwtlV benzene solution of polystyrene (commercially available, manufactured by Wako Pure Chemical Industries),
In addition to coating according to the method described above, a column for evaluation was prepared and evaluated using human blood. The results are shown in Table 7.

Table 7 °-γ Example 3 As the water-soluble material, methyl methacrylate-divinylbenzene copolymer (same as in Example 1) was used, and as the coating agent, 2-hydroxyethyl methacrylate and N, Graft copolymer with N'-diethyl-N-(4-vinylphenethyl)ethylenediamine (content of 2-hydroxyethyl methacrylate 90 mol
Using a 2% wt/V methanol solution of
It was evaluated using human blood. The evaluation results are shown in Table 81C, and the outline of the materials used is shown in Table 9.

Table 8 Table 9 Comparative Example 3 As a water-insoluble material, glass particles (particle size 350-500
The same as in Example 3 except that μ, the same as in Comparative Example 1 of Example 1) was used. The results are shown in Table 10.

Table 10 Example 4 Average spherical diameter 10 obtained by suspension polymerization as a water-insoluble material
The same conditions as in Example 1 were used except that particles of methyl methacrylate-thyl methyl acrylate copolymer (96:4% by weight) of 0μ, 400μ, and 800μ were used.

Table 11 shows the results.

Table 11 Example 5 Potyl methacrylate divinylbenzene copolymer (same as Example 1) with a sphere diameter of 350 to 500 μm was used as the water-insoluble material, and the coating agent was %2-
A 2% wt/V methanol bath solution of a copolymer of hydroxyethyl methacrylate and dimethylaminoethyl methacrylate was used and coated according to the method described above to prepare an evaluation column, which was evaluated using human blood. Table 12
The results are shown in

Table 12 Contact angle of 2-hydroxyethyl methacrylate-dimethylaminoethyl methacrylate copolymer: 18±4
Comparative Example 4 The same conditions as in Example 3 were used except that a 2-hydroxyethyl methacrylate polymer was used as the coating agent. The results are shown in Table 13.

Table 13 Contact angle of 2-hydroxyethyl methacrylate polymer; 17 ± 3 (degrees) Comparative Example 5 As an example of a material used for an artificial liver, activated carbon manufactured by Kenbetsu Kagaku (particle size 350 ~500μ)
Using a 2% wt/'V methanol solution of a polymer of 2-hydroxyethyl methacrylate as a coating agent, coating was performed according to the method described above, and an evaluation column was prepared and evaluated using one human blood. . Table 141C shows changes in platelet passage over time and average values.

Table 14 All samples of Examples 1 to 5 were used to observe column population and platelet morphological changes in blood at the outlet side using a microscope. I couldn't recognize it.

(Effects of the Invention) As is clear from the above explanation, the extracorporeal circulation treatment base material of the present invention has a remarkable effect on the viscosity and activation of thrombotic substances such as platelets, compared to conventional whole blood perfusion materials. It is a base material with good blood compatibility.

Furthermore, since there are no safety issues such as eluates, it can be more safely applied to the human body.

Since it is possible to have a molecular weight of 1000 to several 1000, it can be used as a material for extracorporeal circulation treatment targeting proteins with a small molecular weight of several 1000 or more. Since it is formed into particles, the surface area is large (the volume of the container filled with the base material is 50 to 4
The target component can be sufficiently removed at about 0-0. Therefore, the priming volume decreases, which reduces the burden on the patient.

Sarak can freely immobilize ligands on the base material of the present invention depending on the purpose of treatment, so it is possible to selectively remove target malignant substances while maintaining good blood compatibility, or to activate immune cells. can do. Therefore, there are no symptoms of decreased platelet count during extracorporeal circulation, and there are no problems such as pressure loss or clogging of the treatment device.

There is no reduction in the removal performance or activation ability of the target component (suitable extracorporeal circulation therapy can be performed).

Based on these facts, the base material of the present invention can be applied to general uses such as purifying and regenerating body fluids such as plasma, blood, and body fluid cells, and activating immune cells, and can be used to treat cancer, immunoproliferative syndrome, etc. , rheumatoid arthritis, collagen diseases such as systemic lupus erythematosus, autoimmune diseases such as myasthenia gravis, allergic diseases such as bronchial exhalation, and diseases and phenomena related to the body's immune system such as organ transplant rejection. It can be effectively used for extracorporeal circulation treatment of skin diseases such as psoriasis and albinism, kidney diseases such as nephritis, and liver diseases such as hepatitis.

In addition, 1 artificial blood vessel, 1 artificial valve, 1 artificial organ, including an artificial heart and heart-lung machine, 1 artificial bone, and 1 artificial joint.

Artificial organs implanted within tissues, such as pacemakers,
Furthermore, 1 is used for medical device materials such as syringes and catheters.
The base material of the present invention can be effectively used.

[Brief explanation of drawings]

The drawing is a sectional view showing an example of the form of a device using a material in which a ligand is immobilized on a base material of the present invention. l...Device 2...Cylinder 3.
3'...- Filter 4.4'... Backing 5... Body fluid inlet 6... Cap 7... Body fluid outlet 8... ... Cap 9 ... Material layer

Claims (1)

[Claims] A substrate for extracorporeal circulation therapy, characterized in that a water-insoluble material having a contact angle in the range of 20 to 85 degrees is coated with a polymer having a nitrogen-containing basic functional group.