JPH01104273A - Body fluid purifying material and apparatus - Google Patents

Abstract

PURPOSE:To adsorb and remove an etiological cause substance by high adsorbing capacity and adsorbing specificity, by immobilizing dipeptide or a derivative thereof on the surface of a water-insoluble carrier. CONSTITUTION:Dipeptide or a derivative used as the ligand in a body fluid purifying material is a compound or a derivative thereof formed by mutually dehydrating two same or different amino acids between the carboxyl group of one of them and the amino group of the other to form an acid amide bond or peptide bond. In a body fluid purifying apparatus 1 having a column packed with the purifying material 5 prepared by immobilizing dipeptide or a derivative thereof on the surface of a water-insoluble carrier, a column container 4 equipped with a fluid introducing port 2 and a fluid lead-out port 3 is packed with the granular body fluid material 5 and said purifying material 5 is held in the column container by the filters 6, 6' provided in the vicinities of the fluid introducing port 2 and the fluid lead-out port 3.

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to a body fluid purification material and a body fluid purification device. Specifically, the present invention relates to a body fluid purification material and a body fluid purification device that adsorb and separate pathogenic substances from body fluids such as blood with high selectivity.

(Prior Art) In recent years, plasma exchange therapy has been used as a treatment for diseases in which specific components in body fluids cause serious symptoms. This therapy is particularly effective in cases where dialysis of the pathogenic substance is difficult, including autoimmune diseases such as myasthenia gravis, rheumatoid arthritis, and lupus erythematosus.
It is indicated for immune-related diseases such as glomerulonephritis, bronchial asthma, and polyneuritis, rejection reactions associated with organ transplants, liver failure, hypertension, and cancer diseases. Among these, autoimmune diseases refer to diseases that occur due to humoral or cellular immune responses against antigens possessed by one's own cells and tissues, resulting in the formation of antibodies against oneself.

However, since plasma exchange therapy indiscriminately removes all plasma components, it not only removes pathogenic substances such as immunoglobulin, fibrinogen, and immune complexes, but also loses albumin and other useful plasma components. There's a problem. In addition, there is a shortage of plasma and plasma products as replacement fluids,
Many problems have been pointed out, including serum hepatitis, AIDS, co-occurrence of allergic diseases, and side effects.

For this reason, a self-plasma purification method that selectively removes large molecular weight proteins such as immunoglobulin, fibrinogen, and immune complexes, which are pathogenic substances, and returns self-purified plasma containing albumin and other useful plasma components to the brain. are being studied and implemented.

Conventionally, adsorbents used in such self-plasma purification methods include porous resins (for example, Amberlite X A D, which is a styrene divinylbenzene copolymer)
-4 [To Amber ite X〇-4], o-
ion exchangers (eg, carboxymethyl cellulose, diethylaminoethyl agarose, etc.), inorganic porous bodies (eg, porous glass, ceramics, etc.), and affinity adsorbents. However, conventionally known porous resins and ion exchangers not only have low adsorption capacity but also have low adsorption specificity, so they also adsorb useful components such as albumin in body fluids.

As a result, when these are applied as adsorbents to the above-mentioned therapy, not only the therapeutic effect is insufficient, but also there are safety problems such as osmotic pressure abnormality, which is characteristic of hypoproteinemia, and edema. In addition, although inorganic porous materials have relatively good adsorption capacity and adsorption properties, they are still insufficient for practical use.

In contrast, affinity adsorbents have excellent adsorption capacity and adsorption properties, and are considered promising as adsorbents for body fluid purification. This type of adsorbent has different adsorption properties,
They are classified into biological affinity adsorbents and physicochemical affinity adsorbents. Biological affinity adsorbents bind to pathogenic substances in body fluids through biological interactions such as antigen-antibody binding, complement binding, and F'c fragment binding.
It adsorbs this and has extremely excellent adsorption specificity. However, most of these use bioactive polymers such as DNA, anti-LDL antibodies, and protein A as ligands (substances that have an affinity for the target pathogenic substance), so raw materials are expensive and difficult to obtain. be. Furthermore, due to the poor stability of the ligand, it is difficult to maintain its activity during the manufacture, sterilization, storage, transportation, and storage of adsorbents and columns packed with them. In addition, since the ligand is essentially a physiologically active substance, it must be taken into account that when it comes into contact with blood, it not only exhibits the desired affinity, but also exhibits its original physiological action and causes side effects. . Moreover, since the ligand is a foreign protein, if it is released and eluted from the adsorbent, side effects will occur due to its antigenicity. On the other hand, physicochemical affinity adsorbents bind to pathogenic substances in body fluids through physical or chemical interactions such as electrostatic bonds and hydrophobic bonds, and remove them by adsorption and separation. As the ligand in this case, for example, synthetic compounds such as polylysine, methylated albumin, tryptophan, and phenylalanine can be used. Therefore, it has advantages such as being able to be manufactured in large quantities, being relatively inexpensive, and having stable activity. Furthermore, since many of them are generally excellent in safety when they come into contact with blood, they are the most promising adsorbent for use in the above-mentioned therapy. Furthermore, in terms of its action and price, it can be used not only as an adsorbent for the above-mentioned therapies, but also for the separation and purification of specific components of body fluids such as immunoglobulins, immune complexes, and immune-related soluble factors, or for the testing of these components. There is expected. Conventionally known such physicochemical affinity adsorbents include porous materials having carboxyl groups or sulfonic acid groups on the surface; JP-A-57-170263, JP-A-57-197294), hydrophilic carriers to which hydrophobic amino acids are bound (JP-A-57-122875, JP-A-57-197294);
-15924, 58 165859, 58-1
65861), hydrophilic carriers to which modified immunoglobulin (IgG) is bound (JP-A-57-77624, JP-A-57-77625, JP-A-57-156035), porous bodies to which methylated albumin is bound. Japanese Unexamined Patent Publication 1973-
120875, JP 55-125872), hydrophilic carriers to which sugars are bound (JP-A-57-134164,
JP 57-192560, JP 58-61752, JP 58-61752;
No. 8-98142), an insoluble carrier to which a peptide having affinity for sugar chains is bound (Japanese Patent Application Laid-Open No. 60-163667)
) etc.

However, these conventional physicochemical affinity adsorbents still cannot be said to have sufficient performance for application in the treatment of autoimmune diseases, etc. by self-plasma purification therapy as described above, and have even higher adsorption efficiency and adsorption specificity. It has been desired to develop a purification material and a purification device that can remove pathogenic substances and have less negative impact on body fluids.

(Problems to be Solved by the Invention) Therefore, an object of the present invention is to provide a novel body fluid purification material and a body fluid purification device. Another object of the present invention is to provide a highly safe body fluid purification material and body fluid purification device that can adsorb and remove pathogenic substances from body fluids such as blood with high adsorption capacity and adsorption specificity. A further object of the present invention is to provide a body fluid purification material and a body fluid purification device that have stable performance and are easy to sterilize, store, transport, and store. Another object of the present invention is to provide a body fluid purification material and a body fluid purification device that are inexpensive and can be manufactured in large quantities.

(Means for Solving the Problems) The above objects are achieved by an aqueous purification material characterized in that a dipeptide or a derivative thereof is immobilized on the surface of a water-insoluble carrier.

The present invention also provides a body fluid purifying material in which the dipeptide or its derivative is a dipeptide of an acidic amino acid and an aromatic ring amino acid or a heterocyclic amino acid, or a derivative thereof. The present invention further provides a body fluid purifying material, wherein the dipeptide or its derivative is a dipeptide of aspartic acid or glutamic acid and phenylalanine, tyrosine, tryptophan, proline or hydroxyproline, or a derivative thereof. The present invention further includes:
This indicates a body fluid purifying material in which the dipeptide or its derivative is aspartyl phenylalanine or its derivative. The present invention further provides a body fluid purifying material in which the dipeptide or its derivative is aspartyl phenylalanine methyl ester. The present invention also provides
This shows a body fluid purification material in which the water-insoluble carrier is a particulate carrier. The present invention further provides that the particulate carrier has an average particle diameter of 0°0.
This shows a body fluid purifying material having a thickness of 5 to 5 mm.

The present invention also provides a body fluid purification material in which the water-insoluble carrier is porous. The present invention also provides a body fluid purifying material in which the water-insoluble carrier is composed of a synthetic organic polymer, a natural organic polymer, or an inorganic substance. The present invention further provides a body fluid purifying material in which the water-insoluble carrier is porous acrylic ester particles or porous chitosan particles. Furthermore, the present invention provides a body fluid purifying material in which a dipeptide or a derivative thereof is immobilized on the surface of a water-insoluble carrier by covalent bonding. The present invention also provides a body fluid purifying material in which a dipeptide or a derivative thereof is covalently immobilized on the surface of a water-insoluble carrier via a spacer.

The above objects are also achieved by a body fluid purification device characterized by having a column filled with a purification material formed by immobilizing a dipeptide or a derivative thereof on the surface of a water-insoluble carrier.

The present invention also provides a body fluid purification device that is thermally sterilized. The present invention also provides a body fluid purification device in which the purifying material is formed by immobilizing a dipeptide or its derivative on the surface of a particulate water-insoluble carrier. The invention also provides a body fluid purification device in which the column is made of polypropylene or polycarbonate. The present invention further provides a body fluid purification device comprising a filter having a mesh between the inlet/outlet of the column and the purification material layer, through which body fluid components can pass, but through which the purification material cannot pass. The present invention also provides a body fluid purification device in which the dipeptide or its derivative is a dipeptide of an acidic amino acid and an aromatic or heterocyclic amino acid. The present invention further provides a body fluid purification device, wherein the dipeptide or its derivative is a dipeptide or a derivative thereof of aspartic acid or glutamic acid and phenylalanine, tyrosine, tryptophan, proline or hydroxyproline. The present invention further provides an immunoadsorption device in which the dipeptide or derivative thereof is aspartyl phenylalanine or a derivative thereof. The present invention further provides a body fluid purification device in which the dipeptide or its derivative is aspartyl phenylalanine methyl ester. The present invention also provides a body fluid purification device in which the particulate water-insoluble carrier has an average particle size of 0.05 to 5 mm. The present invention further provides a body fluid purification device in which the water-insoluble carrier is porous.

The present invention also provides a body fluid purification device in which the water-insoluble carrier is composed of a synthetic organic polymer, a natural organic polymer, or an inorganic substance. The present invention further provides a body fluid purification device in which the water-insoluble carrier is porous acrylic ester particles or porous chitosan particles. The present invention also provides a body fluid purification device in which a dipeptide or a derivative thereof is covalently immobilized on the surface of a water-insoluble carrier. The present invention also provides a body fluid purification device in which a dipeptide or a derivative thereof is covalently immobilized on the surface of a water-insoluble carrier via a spacer.

Incidentally, the pathogenic substance to which the body fluid purification material and body fluid purification device of the present invention is adsorbed is a harmful protein contained in body fluids, and in more detail, ordinary immunoglobulin (IgA).

IgD, IgE, IgG, IgM), rheumatoid factor, or anti-nuclear antibody, anti-DNA antibody, anti-lymphocyte antibody, anti-erythrocyte antibody, anti-platelet antibody, anti-acetylcholine receptor antibody, anti-colon antibody, anti-renal glomerulus/pulmonary base Membrane antibodies, desmosomal antibodies between epidermal cells, anti-insulin receptor antibodies, anti-minilin antibodies, anti-haemophilia factor II antibodies and other autoantibodies, reduced products of immunoglobulins, between immunoglobulins or between immunoglobulins and other substances These include complexes with antigens and antigen-like substances (particularly antigens and antigen-like substances), lymphocytes, complement, fibrinogen, and the like.

(Function) Therefore, the body fluid purifying material according to the present invention is a type of physicochemical affinity adsorbent characterized by immobilizing dipeptides or derivatives thereof on the surface of a water-insoluble carrier. As shown, it exhibits extremely high adsorption efficiency and adsorption specificity for pathogenic substances.

In general, purification materials composed only of hydrophobic compounds, that is, purification materials with strong hydrophobic binding, are selectively removed because not only pathogenic substances but also many useful proteins such as albumin fractions in body fluids adhere to them. Not suitable for On the other hand, purification materials made of hydrophilic compounds with strong hydrogen bonding properties or compounds with many charged groups with strong electrostatic bonding properties cannot sufficiently adsorb pathogenic substances because the amount of protein attached is extremely low. Considering these points, in order to separate and remove only pathogenic substances in body fluids with high adsorption efficiency and adsorption specificity, it can be said that a purifying material that has a suitable combination of various interactions is suitable.

The body fluid purifying material of the present invention contains the above-mentioned pathogenic substances, especially
Immunoglobulin or immunoglobulin-related compounds (hereinafter referred to as immunoglobulin compounds).

), the hydrophobic and electrostatic properties of the purifying material exert interaction forces with the amino acid residues of the adsorbed portion of the adsorbed compound (immunoglobulin compound), and the three-dimensional structure of the surface of the purifying material It can be interpreted that the favorable results were obtained because the adsorbed compound (immunoglobulin compound) was appropriately applied to the intramolecular pocket.

In addition, the compound used as a ligand in the body fluid purification material of the present invention is a dipeptide, and unlike the physiologically active polymers used in conventional biological affinity adsorbents, it can be easily synthesized or synthesized at a relatively low cost. It is readily available and has excellent stability.

Therefore, the production, sterilization, storage, transportation, storage, etc. of the body fluid purification material and body fluid purification device according to the present invention are all extremely easy, and the activity of the purification material is not reduced or lost in these conditions. Furthermore, regarding safety when it comes into contact with body fluids, even if dipeptides, which are ligands, are released from purifying materials, dipeptides do not exhibit physiological effects in body fluids and are easily degraded and metabolized as amino acids. Therefore, it is extremely good.

Hereinafter, the present invention will be explained in more detail based on embodiments.

The body fluid purifying material of the present invention is characterized in that a dipeptide or a derivative thereof is immobilized on the surface of a water-insoluble carrier.

The dipeptide or its derivative used as a ligand in the body fluid purification material of the present invention is a dipeptide or a derivative thereof that is formed by dehydrating two amino acids of the same type or of the same type between one carboxyl group and the other amine group to form an acid amide bond, that is, a peptide. It is a compound formed by combining or a derivative thereof. Such dipeptides or derivatives thereof are preferably aspartic acid, glutamic acid,
Acidic amino acids such as asparagine, glutamine, etc. and aromatic ring amino acids such as phenylalanine, tyrosine, short tyrosine, surinamine or tryptophan,
Dipeptides with heterocyclic amino acids such as proline, hydroxyproline, histidine, etc. or derivatives thereof, particularly preferably aspartic acid or glutamic acid with 722-alanine, tyrosine, tryptophan, etc.
It is a dipeptide with proline or hydroxyproline or a derivative thereof. Among these, more preferred dipeptides or derivatives thereof include aspartyl phenylalanine, which is a dipeptide of aspartic acid and phenylalanine, and its derivatives, and aspartyl phenylalanine methyl ester is particularly preferred.

Such dipeptides interact with pathogenic substances, especially immunoglobulin compounds, due to the hydrophobicity of the main part of the compound and the electrostatic properties of heteroatoms that interact with the amino acid residues of adsorption sites in the molecular chain of the pathogenic substance. It is thought that the high affinity of the non-antigen/antibody system was obtained because each of them exerted an interaction force and, in addition, the three-dimensional structure of the compound appropriately fit into the intramolecular pocket of the pathogenic substance.

In the body fluid purification material of the present invention, the water-insoluble carrier on which such organically synthesized ligands are immobilized may be hydrophilic or hydrophobic, such as agarose-based, dextran, etc. cellulose-based, polyacrylamide-based, polyvinyl alcohol-based, polyvinyl and lolidone-based, polyacrylonitrile-based, styrene-divinylbenzene copolymer, polystyrene-based, acrylic ester-based, methacrylic ester-based, polyethylene-based, polypropylene-based, poly 4-fluoroethylene system,
Organic polymers such as ethylene-vinyl acetate copolymer system, polyamide system, polycarbonate system, polyvinylidene fluoride system, polyvinyl formal system, polyarylate system, polyether sulfone system, glass system, alumina system, titanium system, activated carbon system, ceramics Collagen, chitosan, etc., which are natural organic polymers derived from inorganic materials or living organisms, may be used. Furthermore, immobilized enzymes and known carriers used in affinity chromatography can be used without any particular limitations. The form of such a water-insoluble carrier is not particularly limited, and various forms such as particulate, fibrous, hollow fiber, membrane, and plate forms can be used, but body fluids such as blood and plasma can be used. From the viewpoint of liquid permeability and ease of handling during preparation of the purifying material, particles are preferably used, particularly those having an average particle size of 0.05 to 5 mm. In addition, the average particle size is calculated as the weight average of the particle size of each grade, which is the intermediate value between the upper limit particle size and the lower limit particle size of each grade after classification using a sieve specified in JI 5-Z-8801. This is what I did. Further, the particle shape is preferably spherical from the viewpoint of not causing physical damage to cells and making it easier to obtain uniform particles. In addition, the water-insoluble carrier preferably has a porous structure, since it can retain a larger amount of the above-mentioned dipeptide or its derivative on its surface, thereby achieving even higher adsorption efficiency. It is desirable that the average pore diameter of the pores of the porous body is 100 to 5,000, particularly 200 to 5,000.
Something in the range to 3000 is desired. When the water-insoluble carrier has a porous structure, the average pore size of the pores is set to 100 to 5,000 mm.If the average pore size is smaller than 100 mm, the penetration and diffusion of pathogenic substances in body fluids into the pores is reduced. On the other hand, if the average pore diameter is larger than 5,000 mm, the strength of the porous material decreases and the surface area decreases, making it impractical. Note that the average pore diameter referred to here refers to the value measured by a mercury intrusion porosimeter, which calculates the amount of pores by injecting mercury into a porous body and calculating the pore size from the amount of mercury that entered, and then calculates the pore diameter from the pressure required for intrusion. It is.

Taking these points into consideration, particularly preferred water-insoluble carriers include porous acrylic ester particles and porous chitosan particles.

In the body fluid purification material of the present invention, any known method such as covalent bonding, ionic bonding, physical adsorption, embedding, or insolubilization by precipitation on the surface of the carrier can be used to immobilize the dipeptide or its derivative on the surface of the water-insoluble carrier. However, in terms of the elution properties of unimmobilized ligands,
It is preferable to use it after being fixed and insolubilized by covalent bonding.

Therefore, in general, known insoluble carrier activation methods and ligand immobilization methods used in the field of immobilized enzymes, affinity chromatography may be preferably used. Furthermore, if necessary, it is also possible to introduce a spacer of arbitrary length between the surface of the water-insoluble carrier and the dipeptide or its derivative as a ligand. The spacer used in this case may be anything that is in the form of a molecule, can be interposed between the surface of the water-insoluble carrier and the ligand, and can have an arbitrary length, and can form a bond between the surface of the water-insoluble carrier and the ligand. Any skeleton structure may be used as long as it does not sterically inhibit the structure. When such a spacer is interposed between the surface of the water-insoluble carrier and the ligand, the fluidity of the spacer and the excluded volume effect prevent body fluid components from approaching and interacting with the surface of the purifying material. As a result, it is thought that the adhesion of albumin fractions, etc., which have a weak interaction with the purifying material, is suppressed preferentially, and the effect of suppressing the non-specific adhesion of these non-pathogenic substances is produced. In the case where a spacer is interposed between the surface of the water-insoluble carrier and the ligand, any known method may be used for immobilization as described above, but it is preferable to use a covalent bond. It is fixed.

The body fluid purification device of the present invention includes a water-insoluble carrier as described above,
Preferably, it is characterized by having a column not filled with a body fluid purifying material formed by immobilizing a dipeptide or a derivative thereof on the surface of a particulate water-insoluble carrier.

In a body fluid purification device having a column filled with this body fluid purification material, the material constituting the column container may be synthetic resins such as polypropylene, polycarbonate, polystyrene, polymethyl methacrylate, etc., or metals such as glass and stainless steel. However, polypropylene, polycarbonate, etc., which can be sterilized by autoclaving and are easy to handle, are particularly preferred. Further, it is preferable that a filter having a mesh that allows body fluid components to pass through but does not allow the purification material to pass is preferably provided between the inlet/outlet of the column of this body fluid purification device and the body fluid purification material layer filled with the body fluid purification material. The filter may be made of any material as long as it is physiologically inert and has high strength, especially polyester,
Polyamide is preferred.

FIG. 1 is a sectional view schematically showing an embodiment of a body fluid purification device according to the present invention. In this embodiment, the body fluid purification device 1 includes a column container 4 having a fluid inlet 2 and a fluid outlet 3 filled with particulate body fluid purification material 5. and is retained in the column vessel by a filter 6.6- provided near the fluid outlet 3. In order to perform extracorporeal blood circulation therapy using this body fluid purification device 1, the body fluid purification device 1 is incorporated into a circuit as shown in FIG. 2, for example.

Blood from a patient is introduced into the circuit from a blood introduction lower, passes through a blood pump 8 and a chamber 9, is supplied at a constant flow rate to a plasma separation device 10, and is separated into blood cell components and plasma components. Plasma drawn out from the plasma outlet 11 of the plasma separation device 10 passes through the plasma pump 12 and the chamber 13 1 and is introduced into the body fluid purification device 1 from the fluid introduction port 2, and then is introduced into the body fluid purification material 5 housed in the column container 4. After being subjected to adsorption treatment, the fluid is led out from the fluid outlet 3. The plasma components processed in the body fluid purification device 1 are led out from the blood cell outlet 15 of the plasma separation device 10 in the mixing chamber 1.4, mixed with the blood cell components again, passed through a thermostatic chamber 16, and then delivered to the patient through the blood outlet 17. is returned to the body of

The body fluid purification device of the present invention is thermally sterilized by filling purified water, physiological saline, etc. into the device and heating the device before use.

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

Example 1 First, glutaraldehyde was dissolved in 0.1M phosphate buffer (pH 7.4) to a concentration of 5 (w/v)%. Chitosan beads (
Chitopearl BCW3003, lot number H-0132, manufactured by Fujibo Co., Ltd., 0°2 to 0.4 g (wet type!)
ml, and after degassing, add the blood mixer (
Stir overnight at room temperature using a BM-101 (manufactured by Kayagaki Irika Kogyo Co., Ltd.). Next, after washing the obtained reaction product with distilled water, O, LM carbonate M solution (pH 10>
, a solution of 05M aspartyl phenylalanine methyl ester (Ajinomoto layer, aspartame) was added and degassed,
Stir overnight at room temperature using a blood mixer. After washing with distilled water, 1M monoethanolamine (pH
8,0> was added, degassed, and stirred overnight in a blood mixer.

After washing this with distilled water, 1 (w/v)% sodium borohydride in 0.1M phosphate buffer (pH 7,4
) solution and allowed to react at room temperature for 3 hours with occasional stirring. This reaction reduced and stabilized the azomethine bond produced by the reaction between the aldehyde group of glutaraldehyde and the amine group of chitosan and aspartyl phenylalanine methyl ester. Furthermore, distilled water, pH 4.0 containing 0.5M sodium chloride,
A cleaning material was prepared by repeatedly washing with 0.02M acetic acid M solution, PHIOlo, and 2M charcoal MM buffer solution, and was used in the following adsorption experiment.

For the adsorption experiment, first, a glass test tube (manufactured by Terumo, Larbo) was used.
Add 1 g of the purification material obtained above (wet nest) and 5 ml of normal human plasma without adding 140 μD/ml of CPD solution as an anticoagulant to the container. After degassing, stir at 37°C using a blood mixer. Hot air circulation thermostat (manufactured by Tabai, P
(S)-212 type) for 90 minutes. After a predetermined period of time, polyester mesh (225
After filtration using bromcresol green (bromcresol green), the amount of albumin and total protein in the solution were measured, respectively.
BCG) method and Biuret method. Furthermore, the amount of globulin was determined as the difference between the amount of total protein and the amount of albumin. That is, for convenience, fibrinogen was also included in globulin in the calculation.

Apart from this, the same operation as above without adding purification material (
blank), and the albumin and globulin levels were measured and calculated in the same manner. The amount of adsorption is calculated by subtracting the amount of albumin and globulin when using the purification material from the amount of albumin and globulin in the blank, respectively, and further by dividing the value when using the purification material by this value. The adsorption rate for each was calculated.

Further, the oxidation protein, gG, and Ig in the furnace liquid obtained in the above operation.
IgG by measuring the amount of M by single radial immunodiffusion (SRID) and subtracting it from the blank value in the same manner as above.
, by dividing the amount of IgM adsorbed by this value, I
The adsorption rates of gG and IgM were determined. These results are shown in Table 1.

Comparative Examples 1-2 0.05M dissolved in 0.1M carbonate buffer (pH 10>
Instead of using a solution of aspartyl phenylalanine methyl ester, use the same <O, 1M carbonate buffer (pH 10
A purifying material was prepared in the same manner as in the example except that a solution of 0.1M aspartic acid (Comparative Example 1) or 0.1M phenylalanine (Comparative Example 2) dissolved in 100% of 0.1M aspartic acid (Comparative Example 2) was used and subjected to the same adsorption experiment. The results are shown in Table 1.

Comparative Example 3 Chitosan beads (manufactured by Fujibo Co., Ltd., Chito Pearl BCW30
03, lot number H-0132> was used as a purifying material and subjected to the same adsorption experiment as in the example. The results are shown in Table 1.

As is clear from the results shown in Table 1, the cleaning material according to the present invention (Example 1), which is made by immobilizing aspartyl phenylalanine methyl ester, which is a dipeptide, on the surface of chitosan beads, which is a carrier, uses amino acids as ligands. Compared to Comparative Examples 1 and 2) and those using chitosan beads themselves as purification materials, IgG was clearly reduced.
It was shown to more selectively adsorb IgM and IgM.

Example 2 A purifying material was prepared in the same manner as in Example 1, except that a dipeptide of glutamic acid and phenylalanine (α-L-glutamyl-L-phenylalanine, manufactured by Kokusan Kagaku) was used instead of aspartyl phenylalanine methyl ester. was used for adsorption experiments. The results are shown in Table 2.

Example 3 A purifying material was prepared in the same manner as in Example 1, except that a dipeptide of aspartic acid and tryptophan (L-tryptophyl-L-asparatic acid, manufactured by Kokusan Kagaku) was used instead of aspartyl phenylalanine methyl ester. , and subjected to similar adsorption experiments. Second result
Shown in the table.

Comparative Examples 4-7 Example 1 except that instead of using aspartyl phenylalanine methyl ester, glutamic acid (Comparative Example 4), aspartic acid (Comparative Example 5), phenylalanine (Comparative Example 6) or tryptophan (Comparative Example 7) was used. A purifying material was prepared in the same manner as above and subjected to the same adsorption experiment. The results are shown in Table 2.

It should be noted that the adsorption experiments on purification materials in Example 1 and Comparative Examples 1 to 3 and the adsorption experiments on purification materials in Examples 2 to 3 and Comparative Examples 4 to 7 were performed on plasma from separate normal individuals. It was done.

The results shown in Table 2 are similar to the results shown in Table 1, and the dipeptide α-L-glutamyl-L-phenylalanine (Example 2) or L-tryptophyl-L-asparatic acid was applied to the surface of the chitosan beads as a carrier. (
The purification material according to the present invention obtained by immobilizing Example 3) shows that it clearly adsorbs IgG and IgM more selectively compared to those using amino acids (Comparative Examples 4 to 7). Met.

(Effects of the Invention) As described above, the present invention is a body fluid purifying material characterized by immobilizing dipeptides or derivatives thereof on the surface of a water-insoluble carrier. It is possible to safely adsorb and remove proteins such as immune-related soluble factors efficiently, with high selectivity, and is therefore extremely suitable as a purification material for adsorbing and removing pathogenic substances from blood and other body fluids and purifying body fluids. It enables the effective treatment of autoimmune diseases, immune-related diseases, etc. by self-plasma purification therapy, and is not only used for such therapeutic purposes, but also for immunoglobulins, immune complexes, immune-related soluble factors, etc. It is expected to be widely applied to the separation and purification of specific components of body fluids, and the testing of these components.Furthermore, because its activity is stable, it can be used for manufacturing, sterilization, storage, transportation, storage, etc. It is easy and inexpensive. Furthermore, in the body fluid purifying material of the present invention, the dipeptide or its derivative is a dipeptide of an acidic amino acid and an aromatic ring amino acid or a heterocyclic amino acid or a derivative thereof, preferably a combination of aspartic acid or glutamic acid and phenylalanine, tyrosine, tryptophan, proline or hydroxyproline. A dipeptide or a derivative thereof, more preferably aspartyl phenylalanine or a number of derivatives thereof is also preferably aspartyl phenylalanine methyl ester, the water-insoluble carrier is a particulate carrier, particularly a particulate carrier having an average particle size of 0.05 to 5 mm, and Particularly preferred are porous acrylic acid ester particles or porous chitosan particles, which have porosity and are made of synthetic organic polymers, natural organic polymers, or inorganic substances. If it is immobilized on the surface of a water-insoluble carrier by a covalent bond, or if it is immobilized by a covalent bond on the surface of a water-insoluble carrier via a spacer, the above-mentioned properties will be even better, and the effect will be even better. can be expected.

The present invention further provides a body fluid purification device characterized by having a column filled with a purification material formed by immobilizing a dipeptide or a derivative thereof on the surface of a water-insoluble carrier.
As mentioned above, it is possible to more easily and efficiently bring the body fluid into contact with the body fluid purification material, which has the excellent properties. It becomes possible to separate, purify, or test substances in a shorter time and more successfully. Furthermore, since the body fluid purification device of the present invention exhibits stable adsorption activity without deteriorating its performance even when subjected to moist heat sterilization treatment, safer treatment or operation can be performed.Furthermore, the column is made of polypropylene or polycarbonate. In addition, if a filter is provided between the inlet and outlet of the column and the purification material layer, which has a mesh that allows body fluid components to pass through but does not allow the purification material to pass through, operations such as sterilization will be easier and more efficient. There is less risk of damaging components in the passing body fluid, and pathogenic substances can be selectively adsorbed and removed more safely and efficiently. Furthermore, in the body fluid purification device of the present invention, the body fluid purification material is formed by immobilizing a dipeptide or its derivative on the surface of a particulate water-insoluble carrier, and the dipeptide or its derivative is an acidic amino acid, an aromatic ring amino acid, or a heterocyclic amino acid. or a derivative thereof, more preferably a dipeptide of aspartic acid or glutamic acid and phenylalanine, tyrosine, tryptophan, proline or hydroxyproline, or a derivative thereof, more preferably aspartyl phenylalanine or a derivative thereof, most preferably aspartyl phenylalanine methyl ester. In addition, the water-insoluble carrier is in the form of particles with an average particle size of 0.05 to 5 mm, is porous, and is made of a synthetic organic polymer, a natural organic polymer, or an inorganic substance. Porous acrylic acid ester particles or porous chitosan particles are particularly preferred, and dipeptides or derivatives thereof are immobilized on the surface of the water-insoluble carrier by covalent bonding or via spacers. If it is, it will exhibit even more excellent characteristics.

[Brief explanation of the drawing]

FIG. 1 is a sectional view schematically showing an embodiment of the body fluid purification device of the present invention, and FIG. 2 is a circuit diagram of an extracorporeal circulation therapy incorporating the body fluid purification device of the present invention. 1... Body fluid purification device, 2... Fluid inlet, 3...
Fluid outlet, 4... Column container, 5... Body fluid purification material, 6, 6-... Filter, -110... Plasma separation device, 14... Mixing chamber, 16... Constant temperature bath .

Claims (6)

[Claims] (1) A body fluid purification material characterized by having a dipeptide or its derivative immobilized on the surface of a water-insoluble carrier. (2) The body fluid purifying material according to claim 1, wherein the dipeptide or its derivative is a dipeptide of an acidic amino acid and an aromatic ring amino acid or a heterocyclic amino acid, or a derivative thereof. (3) The dipeptide or its derivative is aspartic acid or glutamic acid, phenylalanine, tyrosine,
The body fluid purifying material according to claim 1 or 2, which is a dipeptide with tryptophan, proline, or hydroxyproline, or a derivative thereof. (4) The body fluid purifying material according to any one of claims 1 to 3, wherein the dipeptide or its derivative is immobilized on the surface of a water-insoluble carrier by a covalent bond. (5) The body fluid purifying material according to any one of claims 1 to 3, wherein the dipeptide or its derivative is covalently immobilized on the surface of a water-insoluble carrier via a spacer. (6) A body fluid purification device comprising a column filled with the body fluid purification material according to any one of claims 1 to 5.