JPS6090039A - Blood purifying adsorbing body - Google Patents

Abstract

PURPOSE:To obtain an adsorbing body not coagulating blood even if adsorbs the malignant substance in body fluids and contacted therewith, by providing an polyanion graft chain and an org. compound having a functional region bondable with a substance to be adsorbed to the surface of an insoluble carrier. CONSTITUTION:A polyanion graft chain and an org. compound having a functional region bondable with a substance to be adsorbed to the surface of an insoluble carrier. By using this blood purifying adsorbing body, even if the malignant substance or cell in body fluids is selectively adsorbed therewith and brougt into contact therewith, blood is not coagulated. The negative change density of the aforementioned polyanion graft chain is pref. 0.5-500mueq/m<2> per a surface area of the insoluble carrier and, as the negatively charged functional group, a sulfonic acid group or a carboxylic acid group is pref. used. In addition, as the functional region bondable with the substance to be adsorbed, for example, there is a homopolymer of mono-, di- and trinucleotide such as adenine used in the adsorptive removal of an antinuclear antibody.

Description

Detailed Description of the Invention The present invention relates to a blood purification adsorbent that removes malignant substances and malignant cells from blood and plasma.
The present invention relates to an adsorbent suitable for whole blood purification. More details include cancer, immunoproliferative disorders, rheumatoid arthritis,
Diseases and phenomena related to the body's immune system, such as systemic lupus erythematosus, allergies, and rejection reactions during transplantation with 1 reduction in benefit;
Or malignant substances or malignant A1 cells that are expressed in body fluids such as blood and plasma in kidney diseases such as gastritis, liver diseases such as hepatitis, etc., and are thought to have WJ-related factors that are closely related to the cause or progression of the disease. Regarding adsorbents for absorption and removal from body fluids〇 Conventionally, as adsorbents for body fluid purification treatment, activated carbon or activated carbon coated with hydrophilic polymers have been used mainly as artificial livers for liver diseases. I've been exposed to it. However, in many diseases, malignant substances and skin cells that are closely related to the cause or progression of the disease are known, and even more selectively than in the whole liquid of the malignant substances and malignant cells. There has been a growing demand for removal, but activated carbon-based adsorbents are
At present, this request cannot be met because the substances that can be adsorbed are limited.

In addition, in body fluid purification treatment using an adsorbent, a major problem has been blood coagulation induced by contact between the adsorbent, which is a foreign substance to the living body, and a body fluid such as blood.

In order to completely prevent this blood coagulation, attempts have been made to coat the blood with l-occupied carbonaceous hydrophilic polymers, but sufficient antithrombotic properties have not been achieved, and heparin, an anticoagulant, has been applied to the blood. blood'#. Currently, coagulation is completely prevented. In this case, since a large amount of heparin is used, there are problems such as side effects such as bleeding in some cases.

As a result of intensive research in response to the demand for selective adsorption and removal of malignant substances, the present inventors have found that the carrier retains a special substance that selectively interacts chemically with the adsorbed substance through chemical bonds. Various adsorbents were discovered and patent applications were previously filed (% Patent Application No. 7152/1982, Japanese Patent Application No. 76776/1982, Japanese Patent Application No. 159444/1982, Japanese Patent Application No. 18923/1983).

``Kienori'' was arrived at as a result of extensive research into the above-mentioned blood coagulation problems regarding the previous invention.

That is, the present inventors, in view of the problem of blood coagulation due to contact between the absorbent and blood based on the prior art as described above,
As a result of intensive research into an adsorbent that selectively adsorbs all of the tragic and malignant cells in body fluids and does not cause blood clots upon contact, we found that the surface of the insoluble carrier contains polyanion graft chains and an adsorbent. The present invention has been completed based on the idea that an adsorbent formed by fully bonding an organic compound (hereinafter referred to as "Pa IJ Gant") having all functional sites capable of binding to an adsorbent substance can solve the above problems. reached.

Is it possible to achieve both platelet adhesion suppression and affinity-type adsorption ability, which were not easily solved in the past? In view of this, it is necessary to bind an organic ligand with affinity for malignant substances and malignant cells in the blood to a carrier, and at the same time provide a means for suppressing platelet adhesion on the same carrier. We have discovered the binding of polyanion graft chains as an excellent means for inhibiting platelets. This is expected to make platelets more difficult to approach due to the electrostatic repulsion of platelets due to their negative charge and the whisker effect (excluded volume effect) on the carrier due to the hydrophilic graft chain, and this effect is actually observed. I was able to confirm that.

As the insoluble carrier in the present invention, when the malignant substance is a soluble body fluid component, it is preferable to use a porous carrier, particularly a porous polymer. The exclusion limit molecular weight (protein) of the porous polymer used in the present invention corresponds to the size of the malignant substance to be adsorbed, and should be at least 20,000 or more to effectively purify blood for the purpose of the present invention. It is necessary to achieve this. The adsorbed substance is an immune complex, especially IgM.
In the case of immune complexes, the number reaches 10 million, so 2 to 1
0 million is preferred. The most common exclusion limit molecular weight for purposes of this invention is 100,000 to 5,000,000.

The specific surface area of the insoluble carrier in the present invention is preferably 577 or more, and more preferably 5571 t/f or more.

There are various methods for measuring the specific surface area, but in the present invention, it was determined by the most common BET method using h'1 elementary gas.

In addition, the sample used for specific surface measurement must be sufficiently dry, but if the carrier is difficult to dry, it should be equilibrated with the water-wet carrier carecetone and then dried under reduced pressure at 60C or less for measurement. It was used as a sample.

The insoluble carrier referred to in the present invention can be in any known shape such as particulate, wood fiber, inner wall filament, membrane, or sheet, but depending on the +tie amount of the organic ligand and the ease of handling as an adsorbent, Particulate or fibrous ones are preferable,
In particular, particles in the form of particles give preferable results.

The particle size of the spherical or particulate carrier is determined by its specific surface area (adsorption capacity as an adsorbent) and body fluid circulation surface, from 50 to 150.
Preferably, the thickness is 0 μm. A more preferable range is 200
~1000 μm.

In addition, when using a fibrous carrier sound, the fiber diameter; 7E
0.02 denier to 10 denier, preferably tL< is in the range of Oj denier to 5 denier. If the fiber diameter is too large, the amount and speed of adsorption of globulin compounds will be reduced, while if it is too small, activation of the coagulation system, blood cell adhesion, and clogging may easily occur.

The above-mentioned insoluble carrier immobilizes the ligand, so the carrier can be activated, and the carrier activation reaction, immobilization reaction,
It is sufficient that the compound is physically safe throughout the entire process including the adsorption operation and can be selected from either hydrophilic compounds or hydrophobic compounds.

Specifically, inorganic-based carriers include activated carbon, glass, etc., and their @conductors, and natural polymer-derived carriers include simple polysaccharides such as cellulose, sepharose, dextran, starch, alginic acid, and chitin. and its derivatives, complex polysaccharides and their derivatives such as agar, pectin, konjac, gum arabic, proteins such as wool and silk proteins, and their derivatives. After that, it is used as a carrier.

1. Regarding synthetic polymers, vinyl polymers include styrene, vinyl acetate, methacrylic ester, acrylic ester, vinyl halide, vinylidene halide, acrylonitrile, acrylamide, methyl vinyl ketone, vinyl pyrrolidone, There are polymers and copolymers of 2-vinylpyridine, ethylene, propylene, butadiene, isoprene, etc. and their derivatives, and ring-opening polymers of cyclic compounds include dimethylcyclopropane, spiro-di-xylylene, norpholyne. , cyclobutene, trioxane, lactide, cyclopolysiloxane, phosphonitrile chloride, polymers and copolymers of N-carboxy-α-amino acid anhydrides and their derivatives, polyformaldehyde, polyethylene oxide, polypropylene glycol, poly-5,3 -bis(chloromethyl)oxacyclobutane, polytetrahydrofuran, polycaprolactam, etc., and derivatives thereof.

Examples of the polycondensate include polyester, polyamide, boria/hydride, polycarbonate, polyurea, polysulfonamide, polyimide, polybenzimidazole, and derivatives thereof.

Examples of resins and others include acrylic resins, methacrylic resins, fluororesins, epoxy resins, urea resins, amino resins, styrene resins, melamine resins, polyurethanes, silicone resins, alkyd resins, and derivatives thereof.

The polymer carrier prepared above can be completely insolubilized by using appropriate comonomers and crosslinking agents as required. , epoxy compounds, dienes, glutaraldehyde, etc. can be selected according to the functional group of the object to be crosslinked (Taiseisha, "Crosslinking agent"),
P5-77゜1981).

Among these insoluble carriers, hydrophilic carriers are generally used because the carrier itself must not non-specifically adsorb biological substances, since the adsorbent is created by utilizing the entire affinity of the ligand immobilized on the carrier. It is preferable that the character is of the same character.

1. Since increasing the immobilization capacity of the ligand is advantageous in increasing the adsorption capacity, it is preferable to use an organic polymer carrier that can easily obtain a high activation rate carrier. Furthermore, it is preferable to use an organic synthetic polymer carrier that can control mechanical and/or hydrodynamic properties more appropriately. That is, a hydrophilic synthetic polymer provides a more suitable carrier. Among such carriers, polymers or copolymers containing vinyl alcohol-/' units are more preferred carriers because of their ease of handling and wide applicability.

In addition, polymer cylinders containing vinyl alcohol units tj coM
In the case of one combination, compounds having triazine groups such as carboxylic acid vinyl ester and triallyl cyanurate and triallyl isocyanurate, and di(meth)acrylic IJ L'-ates such as ethylene glycol dimethacrylate and diethylene glycol dimethacrylate. , polyvinyl ethers such as butanediol divinyl ether, diethylene glycol divinyl ether, and tetravinylglyoxal, and polyallyl ethers such as diarylidene pentaerythritol and tetraallyloxyethane. An insoluble carrier obtained by transesterifying or saponifying the copolymer is preferred from the viewpoint of strength, micropore structure, and chemical stability.

The polyanion graft chain retained on the carrier of the present invention exhibits a negative charge in a neutral electrolyte such as blood or body fluid while bound to the carrier. In the present invention, the negative charge density of the polyanion graft chain is at least 0.02 μeq/rrt per surface area of the insoluble carrier, which can be expected to have the effect of reducing the total interaction with platelets. Furthermore, the preferred range of negative charge density is 0.2, which exhibits excellent antithrombotic properties and suppresses endogenous coagulation activation to a low level.
μeq/ba to 2500 μeq/ba, more preferable range is 0.5 μeq/Tt to 5 otte
q/rn”.

In the present invention, in the case of a multiphase structure in which negative charges are localized on the carrier surface, the negative charge density in the surface layer may be within the above range.

The molecular weight of the polyanion graft chains retained on the carrier of the present invention is preferably 600 Daltons or more on average in number, due to the excluded volume effect of the graft chains.

1, the molecular range that can provide a more stable platelet mucus layer suppression effect and not hinder the adsorption ability is 1.50.
It ranges from 0 to 1,000,000 Daltons.

To illustrate the above polyanion graft chain, as the negatively charged functional group, a sulfonic acid group, a carboxylic acid group, a phosphoric acid ester group, etc. are preferably used, and to give specific examples,
Polygalaccuronic acid, phosphorylated mannan, chondroitin, chondroitin sulfate A% chondroitin C,
Acidic polysaccharides such as hyaluronic acid, alginic acid, pectic acid, and fucanic acid, and those based on natural molecule molecules include carboxymethyl cellulose, 4A+Af2 cellulose, carboxymethyl starch, cellulose phosphate, etc., and their derivatives. Those obtained from polyacrylic acid, polymethacrylic acid, polyvinyl sulfuric acid, polystyrene sulfonic acid, maleic acid copolymer, polyphosphoric acid, polyvinyl phosphoric acid, polystyrene phosphoric acid, polyglutamic acid, polyaspartic acid yHs polyphosphate ester, etc. Although there are derivatives thereof, the present invention is not limited to any polyanionic polymer and is not limited to the above substances.

The organic ligand to be supported on the carrier 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. , RNA, and other nucleic acids, as well as nucleobases, sugars, oligosaccharides, and the like can be used. Also, D present in the blood
N.A.

R) For adsorption and removal of IA and ENA, anti-nucleic acid antibodies such as anti-mercury DNA antibody, anti-dimercury DNA antibody, anti-RNA antibody, and anti-ENA antibody, methylated albumin actinomycin D
Basic compounds such as the following can be used. Furthermore, for the adsorption and removal of immune complexes in the blood, antibodies that imprint on immune complexes such as sleeve components such as CIq, specific proteins such as protein A, anti-heavy chain constant region phase 2 antibodies, and rheumatoid factor etc. can be used.

For the treatment of rheumatoid arthritis and rheumatoid arthritis,
Modified γ-globulins modified by chemical denaturation (aggregation) methods such as urea, guanidine hydrochloride, mercaptoethanol, surfactants, and specific solvents, and physical denaturation (aggregation) methods such as heat, ultrasound, and gas bubbling; Denatured immunoglobulin, aggregated γ-globulin, aggregated immunoglobulin, Fc part of immunoglobulin, heavy chain constant part 2 phase of immunoglobulin, and their modified products by the above-mentioned denaturation methods, antigen-like substances against rheumatoid factor, antirheumatoid factor Antibodies and hydrophobic organic compounds such as tryptophan and phenylalanine can be used. For removal of rheumatoid dead ash complexes, antibodies against immune complexes such as complement components such as C1q, specific proteins such as protein A, anti-heavy chain constant region phase 2 antibodies, and rheumatoid factor can be used.

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

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

Glomerular basement membrane components for the treatment of glomerular gastritis, platelet membrane components and platelet granule fraction components for the treatment of idiopathic thrombocytopenic purpura, and transcortisin and anti-cortiscin antibodies for the treatment of Cushing syndrome. Can be used.

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

For the treatment of hypertension, anti-angiotensin antibody can be used, and for the treatment of hyperlipidemia, heparin and anti-riboprotein antibodies can be used.

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. Can be used.

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

For the treatment of breast cancer and the like, antibodies against protein A, ltt: immunoglobulin antibodies and immunosuppressive factors 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 A5 phytohemaagglutinin, nucleic acids, amino acids, lipids, glycolipids, protamine, heparin, antigens, Known substances capable of binding to adsorbed substances such as antibodies, enzymes, substrates, coenzymes, and glycoproteins can be used.

Furthermore, the carrier of the present invention can be used with two or more kinds of glue cants held therein. Furthermore, two or more types of carriers holding ligands can be used in combination.

In the present invention, the method for immobilizing a polyanion graft chain and an organic ligand on the surface of an insoluble carrier includes covalent bonding,
Any known method can be used, such as ionic bonding, physical adsorption, embedding, or precipitation insolubilization on the surface of the carrier.
Considering the elution properties of these compounds, it is preferable to use them after being immobilized and insolubilized by covalent bonding. For organic ligands, conventional immobilized enzymes, known methods for activating insoluble carriers used in affinity chromatography, and methods for binding to ligands can be used. Furthermore, if necessary, a molecule (spacer) of any length can be introduced between the insoluble carrier and the ligand.

Examples of functional groups that can form covalent bonds include hydroxyl group, amino group, carboxyl group, epoxy group, chloroformyl group, azide group, formyl group, bromoacetyl group, incyanato group, cyanide group, acid chloride group, and acid anhydride group. However, if these functional groups have low reactivity and it is difficult to form all covalent bonds directly with the ligand, highly reactive functional groups can be activated using known activation methods. After conversion, a covalent bond may be formed with the ligand. For example, the hydroxyl group of the carrier can be activated by a cyanogen halide method, an epichlorohydrin method, a bisepoxide method, a halotriazine method, a bromoacetyl bromide method, an ethyl chloroformate method, a carbonyldiimidazole method, and the like. These functional groups can react with nuclear reactive groups such as amine groups, hydroxyl groups, carboxyl groups, and thiol groups of the 4'' ligand to form covalent bonds.

On the other hand, methods for bonding polyanion graft chains to insoluble carriers include, in addition to the method similar to the above-mentioned IJ cant, oxidation graphs using oxygen, ozone, cerium salts, etc. Examples include craft polymerization using peroxides such as nitrile and benzoyl peroxide, radiation graft polymerization, and other graft polymerizations of anionic monomers on carriers. The method of coupling is discussed.

The method for producing an adsorbent according to the present invention comprising the above elements is as follows:
It does not specify the order in which its constituent elements are combined. Specifically, a carrier or adsorbent obtained by introducing a polyanion graft chain into a carrier or adsorbent that does not have a polyanion graft chain also represents one embodiment of the present invention.

Furthermore, the bonding order of the polyanion graft chain and the organic ligand can also be selected depending on the ease of production. In addition, in the method of introducing an organic ligand, all the objectives can be achieved by combining the organic ligand with a monomer and performing polymerization, or by performing simultaneous reactions during the formation of an insoluble carrier by post-crosslinking of a polymeric substance.

That is, the present invention is not dependent on the manufacturing method, as long as the polyanion graft chain and the organic ligand are present in the adsorbent.

The adsorbent of the present invention can be used by being filled and held in a container equipped with an inlet and outlet for body fluids.

The drawing shows an example of a sorption device using the adsorbent of the present invention, in which a body fluid 5 is injected into the opening of one end of a cylinder 2 through a backing 4' (with a filter 3 on the 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 discharge lower part is screwed into the other end opening of the cylinder 2 through a backing 4' with a filter 3' stretched inside to form a container. The adsorbent layer 9 is formed by filling and holding an adsorbent in the gap between 3 and 5'.

The absorbent/body layer 9 may be filled with the adsorbent of the present invention alone, or may be mixed or laminated with other adsorbents. As other adsorbents, for example, activated carbon having a wide range of adsorption capacities can be used.

As a result, a wide range of clinical effects can be expected due to the synergistic effect of the adsorbent. The appropriate volume of the adsorbent layer 9 is about 50 to 400 m1 when used for extracorporeal circulation.

When using the apparatus of the present invention for external circulation, there are two methods as shown below. One method is to pass blood taken from the body directly through a nuclear device for purification, and the other is to purify blood taken from the body using a centrifuge or a model plasma separator. This is a method in which specific components in blood are separated, and then malignant substances and malignant cells contained in those components are removed. At this time, the method for passing body fluids may be determined depending on clinical necessity, or depending on the equipment status of the facility.
The liquid may be passed continuously or intermittently.

As described above, the adsorbent of the present invention can be applied to general uses for purifying and regenerating body fluids such as autologous blood and autologous plasma, and can be applied to cancer, immunoproliferative syndrome, rheumatoid arthritis, Collagen diseases such as systemic lupus erythematosus, autoimmune diseases such as myasthenia gravis, allergic diseases such as bronchial asthma, diseases and phenomena related to biological immune system such as organ transplant rejection, psoriasis, For extracorporeal circulation treatment of skin dandruff diseases such as aging, kidney diseases such as nephritis, liver diseases such as hepatitis, blood diseases, etc., without sacrificing the adsorption capacity, which is the basic property of adsorbents, it has high performance. It is a blood purification adsorbent that can be used to purify whole blood by direct extracorporeal circulation, as it shows excellent antithrombotic properties.

Embodiments of the present invention will be explained in more detail with reference to Examples below.

Example 1 Vinyl acetate 100f, triallyl isocyanurate) 41
y, ethyl acetate 1002, polyvinyl acetate (polymerization degree 50
0) A homogeneous mixture of 7@ and 3.62% of 2,2'-azobisisobutyronitrile, 1% by weight of polyvinyl alcohol, 0.05% by weight of sodium dihydrogen phosphate dihydrate, and phosphorus. Disodium oxyhydrogen dodecahydride 1
．． 5 weight - 0.4 t of total dissolved water into a flask,
Suspension polymerization was carried out by heating and stirring at 65C for 18 hours and then at 75U for 5 hours to obtain a granular copolymer. This polymer was hydrolyzed with caustic soda to obtain a polyvinyl alcohol cross-polarized polymer. The average particle size of this polymer is 350 μm,
It had a surface area of 120 m'/V and a protein exclusion limit molecular weight of 750,000. Using this polymer, all organic ligands were introduced by the following method. That is, the dried carrier 15f was suspended in a solution consisting of all dimethyl sulfoxide iaome and hyoepichlorohydrin 120td, 50% sodium hydroxide aqueous solution 15-1 was added, and the mixture was stirred at 60C for 75 hours to form an epoxy activated carrier. Obtained.

The epoxy group binding site of the gel is 6 per tnt of gel.
It was 5 μmol. Using the epoxy binding gel, L-
An adsorbent was created by binding phenylalanine. Phenylalanine f 0.025 mmoA/d dissolved in p) 19.8 carbonate buffer rffl
'r was prepared, added to the epoxy bonded glue at a ratio of 40 to 20, and reacted at 50C for 20 hours.

Excess active groups were blocked with 1 moz/l glycine. The amount of phenylalanine immobilized was calculated from the absorption at 257.5 nm of phenylalanine remaining in the reaction solution. The amount of phenylalanine bound to the adsorbent is 31μ
It was moz/-. The adsorbent was thoroughly washed with water, then washed with physiological saline, dehydrated, and used as an evaluation test subject.

Further, this adsorbent was taken as 10-ml, and after freeze-drying, styrene sulfonic acid nograft polymerization was carried out on the hydroxyl groups of the adsorbent using ceric ammonium nitrate salt by a conventional method.

The reaction is styrene sulfonic acid 0,145nol/t
%cerium nitrate salt 10 mmol/L total water 250
It was dissolved in πe and mixed with the adsorbent and stirred for 3 hours at 35C.

The negative charge density of the obtained polyanion was measured by cation exchange capacity measurement and was found to be 6.0 μeq/rrt.

The polyanion graft chain of this adsorbent was prepared in the form of a sodium salt by a conventional method and subjected to evaluation.

This adsorbent f 3 me column (L/D = 5) was packed with fresh bovine whole blood (heparin added 500 U/100 ml).
dish liquid) k 1. The solution was passed for 1 hour each using a single pass method using Ome/mix. As a result, no decrease in packing volume, clogging, or decrease in mud storage was observed, and the change in pressure H1 in front of the column was 10 to 20 uuuHg.

The adsorption experiment was carried out by mixing chronic rheumatism patient plasma 3 and adsorbent 1 and incubating at 37C for 2 hours. Analysis of rheumatoid factor is performed using adsorption reaction supernatant, RA) I
A test (RAB number) [manufactured by Fujirebio Co., Ltd.] and R
Using the A (KW) kit [manufactured by Kyowa Pharmaceutical Co., Ltd.],
Positive or negative results were determined based on the presence or absence of an agglutination reaction caused by mixing the patient's plasma with a serially diluted solution, and the antibody titer was calculated based on the highest dilution that showed positivity. The antibody titer of the patient's plasma was RA160 and RAHAi280, and the antibody titer of the adsorbent according to this example was 20.80.

On the other hand, a similarly prepared adsorbent without polyanion graft chains resulted in a decrease in platelets. Moreover, the antibody titer to which phenylalanine was not bound was RA80 and RAHA320 in the same patient's plasma, and good adsorption ability for equine factor was not obtained.

Example 2 As a vinyl compound-based porous polymer, an experiment was conducted using a terpolymer consisting of bitroxylethyl methacrylate-ethylene glycol dimethacrylate-glycidyl methacrylate with an average particle size and an average pore size of 350 μm. I did it. The epoxy density in the polymer is 1 mo1
%. As a specific ligand, β-(p-aminophenyl)-ethylamine was reacted with D-(+)-lactose (4-0-β-D-galactopyranosyl-D-glucose). phenyl)-ethylated D-
(+)-Lactose was used, and the polyanion graft chain was obtained by radical telomerization of acrylic acid hetamine and 2-aminoethanethiol and had an average molecular weight of so, o o oO. (Journal of the Chemical Society of Japan, 1977, (1), p. 88-92).

Polymer 20- was competitively reacted with the former organic ligand 20 (My) and the latter one-terminal amine polyacrylic M1.orv, each with 2.7 m9/7 resin and 3.

An adsorbent gold containing 1R9/ml of resin was obtained.

The negative charge density of the polyanion graft chain at this time is 1.
The average molecular weight was 5 μeq/g, and the average molecular weight was s, o oa daltons. In the same manner as in Example 1, all column flow tests using fresh bovine whole blood were carried out, and no thrombus occurred, and the pressure gauge in front of the column was below s mmHg, indicating stable flow i'L.
showed that.

In the adsorption experiment, 5 volumes of systemic lupus erythematosus patient plasma and 1 volume of adsorbent were mixed and incubated for 3 hours. The amount of anti-T cell antibodies in the plasma after adsorption was measured by fluorescent antibody method using a total normal T cell detector. The measurement operations and conditions are as follows.

1. Separate lymphocytes from peripheral blood of healthy individuals by specific gravity centrifugation, and separate into T cells by nylon wool column method.
Lymphocyte Function Search Method” Chugai Igakusha.

p15. P51 (1980)).

2. Add 0.1 di of plasma after the adsorption experiment to I x 10'' of isolated healthy human T cells and treat with 'U% for 1 hour.

3. After cleaning, apply FITC-labeled anti-human immunoglobulin rabbit antiserum and count all fluorescence-positive cells.

4. Express this as a percentage of fluorescence-positive cells.

As a result, the patient's plasma before adsorption was 23.6%, and the plasma treated with this adsorbent was 0.3%. In addition, the amount of globulin and alpine in each plasma was measured using the iA/G test (A/GB test, Wako pure line), but there were no major differences.

On the other hand, the amount of anti-T cell antibodies in plasma treated with an adsorbent containing only polyanion graft chains that do not bind the organic ligand D-(+')-lactose was 22.8 cm, indicating that the anti-lymphoid antibodies of systemic lupus erythematosus Almost no spherical antibodies were adsorbed. Additionally, D-
With the adsorbent binding only (+)-lactose, a decrease in platelets was observed in a column flow test using fresh blood.

[Brief explanation of the drawing]

The drawing is a sectional view taken along line-j, showing an example of the form of an adsorption device using the blood purification adsorption body of the present invention. 1... Adsorption device 2... Circle f*3 6
＋ ”...Filter 4.4'...Backing 5...Body fluid inlet 6...Cap 7...Body fluid outlet 8. ... Cap 9 ... Adsorbent layer

Claims (1)

[Claims] A blood purification adsorbent characterized in that a polyanion graft chain and a shoulder compound having a functional site capable of binding to an adsorbed substance are bonded to the surface of an insoluble carrier.