JPH03236857A - Adsorbing material for medical care of purifying body fluid - Google Patents

Abstract

PURPOSE:To singularly adsorb self-antibodies and/or immune complex bodies at high activity and maintain stable activity and easily carry out operation of sterilization while providing safety by combining to an insoluble carrier a specific origomer in a specific amount, which origomer contains a specific hydrophobic amino acid or its derivative and/or another hydrophobic amino acid or its derivative. CONSTITUTION:An adsorbing material for medical care of purifying body fluids is composed of an origomer containing a hydrophobic amino acid or its derivative whose solubulity in physiological salt water is less than 100mmol/dl at 25 deg.C and having a molecular weight of less than 1000 and being combined to an insoluble carrier by 0.1 to 30mg per 1ml of the insoluble carrier. A compound whose solubility in physiologi cal salt water is above 100mmol/dl becomes too high in its hydrophilic property and its affinity for self-antibodies and/or immune complex bodies is lowered. When the amount of the hydrophobic amino acid or its derivative being held is below 0.1mg/ml the amount of self-antibodies and/or immune complex bodies is extremely lowered, while the amount of the hydrophobic amino acid or its derivative in excess of 30mg/ml is not preferable because then the adsorption specificity of the adsorbing material is lowered.

Description

Detailed Description of the Invention The present invention is an adsorption system for body fluid purification treatment that specifically adsorbs and removes autoantibodies and/or immune complexes that are thought to be closely related to various diseases caused by the immune system of the body. Regarding materials.

As is well known, autoantibodies and/or immune complexes expressed in the blood are associated with autoimmune diseases such as cancer, immunoproliferative syndrome, rheumatoid arthritis, and systemic lupus erythematosus, as well as allergies, rejection reactions during organ transplants, etc. It is thought that there is a close relationship with the cause or progression of diseases and phenomena related to the body's immune function.

Therefore, by specifically adsorbing and removing the above-mentioned autoantibodies and/or immune complexes from body fluid components such as blood and plasma, it is possible to prevent the progression of the above-mentioned diseases, alleviate symptoms, and further accelerate healing. That was expected.

Conventionally, adsorbents that can be used for this purpose include adsorbents in which protein A is immobilized on an insoluble carrier, acrylic acid ester-based porous resins (e.g., XAD-7, manufactured by Rohm and Haas), carboxymethyl cellulose, etc. Cation exchangers have been proposed.

However, although protein A immobilized on an insoluble carrier has specific adsorption ability for autoantibodies and/or immune complexes, it is difficult to secure raw materials because it is a bioactive protein derived from Staphylococcus aureus. The disadvantage is that the product costs are high, and since the activity is unstable, it is easily deactivated due to handling during immobilization, storage after immobilization, etc. Furthermore, it is difficult to use when in contact with body fluids. In this case, there is a risk of harmful effects due to protein A elution, and in addition, there is a problem that it is difficult to sterilize while suppressing deactivation.

In addition, acrylic acid ester-based porous resins and carboxymethyl cellulose have the disadvantages of low adsorption capacity and low adsorption specificity.Furthermore, they also adsorb albumen in body fluids, so they can prevent abnormalities in osmotic pressure. Therefore, it was impossible to use it as a safe treatment device.

In view of the problems of the adsorbent for autoantibodies and/or immune complexes based on the prior art as described above, an object of the present invention is to provide a material that can be generally used, and which is highly active and capable of absorbing autoantibodies and/or immune complexes. To provide an adsorbent that specifically adsorbs, maintains stable activity, is safe, and can be easily sterilized, and is suitable for body fluid purification or regeneration, and an adsorption device using the same. It is something.

As a result of extensive research in line with the above objectives, the present inventors bound various compounds to insoluble carriers and evaluated their binding activity towards autoantibodies and immune complexes, and found that, quite surprisingly, The present inventors have discovered that a hydrophobic amino acid or its derivative bound to an insoluble carrier hardly adsorbs albumin, but adsorbs autoantibodies and immune complexes with extremely high activity and specificity, leading to the completion of the present invention.

That is, the present invention provides a hydrophobic amino acid or its derivative whose solubility in physiological saline at 25° C. is 100 mmol/d1 or less and/or an oligomer containing the hydrophobic amino acid or its derivative, which has a molecular weight of 1000. The present invention relates to an adsorbent for body fluid purification treatment of autoantibodies and/or immune complexes, characterized in that the following is bound to 0.1 to 30 μm per insoluble carrier IH1.

The adsorbed substances targeted by the present invention are autoantibodies and/or immune complexes, but to be more specific, rheumatoid factors, antinuclear antibodies, anti-DNA antibodies, antilymphocyte antibodies, antierythrocyte antibodies, antiplatelet antibodies, etc. Antibodies, autoantibodies such as acetylcholine receptor antibodies, serum demyelinating antibodies, anti-thyroglobulin antibodies, anti-microseam antibodies, and anti-colon antibodies, immunoglobulin derivatives such as reduction products of autoantibodies, chemical modification products, and inter-autoantibody or self-antibodies. These include complexes of antibodies and other substances, especially antigens and antigen-like substances. Among these, those that are particularly preferable as adsorption substances targeted by the present invention are:
Autoantibodies and immune complexes are closely related to the cause and progression of autoimmune diseases.

The hydrophobic amino acid or derivative thereof used in the present invention is
Solubility in physiological saline less than 100 mmol/d1 (25°
C), more preferably 30 mmol/d or less. A compound with a solubility in physiological saline greater than 100' or limole/d1 becomes too hydrophilic and has a reduced affinity for autoantibodies and/or immune complexes, resulting in an extremely reduced adsorption capacity. Furthermore, an affinity for albumin, which is more hydrophilic, is generated, and albumin is also non-specifically adsorbed, which is undesirable.

Hydrophobic amino acids and their derivatives are defined by Tanford
, Nozaki (J, Am, Chem, Soc,,
1fil-4240 (1962), J. Biol, Ch.
em, 246 2211 (1971)) (Tanford, Nozaki (Journal of American
Chemical Society, 4 Rainbows 4240 (1962),
Journal of Biological Chemistry".

2211 (1971)) refers to amino acids and derivatives thereof having a content of 1500 cal/mol or more and a solubility in physiological saline of 100 mmol/mole or less. Examples include lysine, valine, leucine, tyrosine, phenylalanine, isoleucine, tryptophan and derivatives thereof. Among these hydrophobic amino acids and their derivatives, tryptophan and its derivatives give particularly good results.

Furthermore, the amino acid can be used without any particular limitation on the 1 and d conformations.

Among hydrophobic achnoic acids or their derivatives, non-dissociable, zwitterionic, cationic compounds more selectively adsorb autoantibodies and/or immune complexes when bound to an insoluble carrier. It is preferable to anionic compounds. Anionic compounds have an affinity for albumin and adsorption of autoantibodies and/or immune complexes, contrary to what might be expected since albumin molecules are more electronegative than autoantibodies and immune complexes. quantity has decreased.

For example, phenylalanine methyl ester, phenylalanine, and tyrosine do not have much difference in their hydrophobicity, but when their amino groups are covalently bonded to an insoluble carrier and their autoantibody adsorption ability is evaluated, phenylalanine methyl ester>
An example of this can be seen in the fact that the adsorption capacity of phenylalanine and tyrosine decreases. Also, non-aromatic compounds showed similar results. In particular, a more hydrophilic compound having a sulfonic acid group as an anionic group had more affinity for albumin than autoantibodies and/or immune complexes, and acted as an albumin adsorbent.

This is presumed to be due to the interaction between the charge of the compound, albumin, and the charge of the microenvironment within the molecule.

Based on the above experimental results, the mechanism of action of the present invention is as follows:
Hydrophobic compounds bound to insoluble carriers and autoantibodies and/or
or hydrophobic interaction forces of immune complexes (Van der
It is thought to be based on the Waals force. In addition, it is presumed that charge amount interaction force (Coulomb force) acts secondarily as a delivery force.

The oligomer containing the hydrophobic amino acid or derivative thereof of the present invention has a molecular weight of 1000 or less. This makes it easier to handle during immobilization and to store after immobilization compared to natural polymers such as protein A (molecular weight 42,000). Furthermore, even when the substance is eluted from the insoluble carrier, oligomers with a molecular weight of 10,000 or less are safe and have negligible antigenicity to living organisms, and can be easily sterilized. The oligomer can be obtained by copolymerizing the hydrophobic compound monomer alone or with other compounds. As the hydrophobic amino acid or its derivative monomer, for example, tryptophan or the like can be used.

The insoluble carrier used in the present invention can be either a hydrophilic carrier or a hydrophobic carrier, but when a hydrophobic carrier is used, nonspecific adsorption of albumin to the carrier sometimes occurs, so a hydrophilic carrier is preferable. gives the desired result.

The shape of the insoluble carrier may be any known shape such as particulate, fibrous, hollow fiber, or membrane, but from the viewpoint of the amount of hydrophobic compound retained and ease of handling as an adsorbent, particulate and fibrous carriers are preferred. Preferably.

As a particulate carrier, the average particle size is 150μ to 2500μ.
The average particle size is preferably in the range of μ
After classification in running water using a sieve specified in Z-8801, the intermediate value between the upper limit particle size and the lower limit particle size of each class is taken as the particle size of each class, and the average particle size is calculated as the weight average. Further, the particle shape is preferably spherical, but is not particularly limited.

If the average particle size is 2,500μ or more, the amount and rate of adsorption of autoantibodies and/or immune complexes will decrease;
If it is less than 0μ, activation of the coagulation system and blood cell adhesion are likely to occur. Particulate carriers that can be used include agarose-based, dextran-based, cellulose-based, polyacrylamide-based, glass-based, and activated carbon-based carriers, but hydrophilic carriers having a gel structure give good results. Furthermore, immobilized enzymes and known carriers commonly used in affinity chromatography can be used without any particular limitations.

Porous particles, especially porous polymers, can also be used as particulate carriers. The porous polymer particles used in the present invention are capable of immobilizing a hydrophobic compound on their surfaces, and have an average pore diameter of 300 to 9,000 pores, more preferably 1,000 to 6,000 pores. For the polymer composition, known polymers that can have a porous structure, such as polyamides, polyesters, polyurethanes, and vinyl compound polymers, can be used, but in particular, porous vinyl compounds made hydrophilic with hydrophilic monomers can be used. Polymeric particles give favorable results.

Preferably, the porous structure has an average particle size in the range of 300 to 9000 pores; however, if the average pore size is too small, the amount of autoantibodies and/or immune complexes adsorbed is small; In some cases, the strength of the polymer particles decreases and the surface area decreases, making it impractical.

The average pore diameter was measured using a mercury intrusion porosimeter.

In this method, mercury is injected into a porous material, and the pore volume is determined from the amount of mercury infiltrated, and the pore diameter is determined from the pressure required for intrusion, and pores of 40 Å or more can be measured. A pore in the present invention is defined as a pore communicating from the surface with a pore diameter of 40 Å or more.

The average pore diameter is the value of r when the dV/d log rO value is maximum, where r is the pore diameter and ■ is the cumulative pore volume measured by a porosimeter.

When a fibrous carrier is used, it is preferable that the fiber shape is in the range of 0802 denier to 10 denier, more preferably 0.1 denier to 5 denier. If the fiber diameter is too large, the adsorption amount and adsorption rate of autoantibodies and/or immune complexes will be reduced, and if it is too small, activation of the coagulation system, blood cell adhesion, and clogging are likely to occur.

Fibrous carriers that can be used include regenerated cellulose fibers,
Generally known fibers such as nylon, acrylic, and polyester can be used.

Hydrophobic amino acids or their derivatives can be immobilized on the surface of an insoluble carrier by any known method such as covalent bonding, ionic bonding, physical adsorption, embedding, or precipitation insolubilization on the polymer surface. Considering the elution properties of amino acids or derivatives thereof, it is preferable to use them after they are immobilized and insolubilized by covalent bonding. Therefore, it is possible to use commonly known methods of activating immobilized enzymes, insoluble carriers used in affinity chromatography, and methods of binding with hydrophobic compounds. Furthermore, if necessary, a molecule (spacer) of any length may be introduced between the insoluble carrier and the hydrophobic anannoic acid or its derivative. For example, the hydroxyl group of agarose and the isocyanate group on one side of hexamethylene diisocyanate are reacted and bonded, and the remaining isocyanate group is combined with the amino group, hydroxyl group, sulfhydryl group, carboxyl group, etc. of a hydrophobic amino acid or its derivative. It can be carried out by reacting and combining. The spacer length is
Particularly favorable results are obtained when the number of atoms contained therein ranges from no spacer to 20.

In the present invention, the amount of hydrophobic amino acid or its derivative bound to the insoluble carrier ranges from 0.1 to 30 μ per 1 d of the insoluble carrier. More preferably 0. It ranges from 5■ to 15■. If the retained amount is less than 0.1 .mu./-, the adsorption amount of autoantibodies and/or immune complexes will be extremely reduced, and if it exceeds 30 .mu./rd, the adsorption specificity will be undesirably reduced.

The autoantibody and/or immune complex adsorption device used in the present invention is one in which the autoantibody and/or immune complex adsorbent described above is filled and held in a container equipped with a body fluid inlet/outlet. It is.

In FIG. 1, ■ indicates an example of an adsorption device for autoantibodies and/or immune complexes used in the present invention, in which a backing 4 with a filter 3 stretched inside is attached to the opening at one end of the cylinder 2. A cap 6 having a body fluid inlet 5 through the cap 6
A camp 8 having a lower body fluid outlet is screw-fitted to the other end opening of the cylinder 2 through a backing 4'' with a filter 3° inside to form a container. The adsorbent layer 9 is formed by filling and retaining an adsorbent in the gap of .degree.

The adsorbent 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, adsorbents for other malignant substances (antigens) such as DNA, activated carbon having a wide range of adsorption capacities, etc. can be used. As a result, a wider range of clinical effects can be expected due to the synergistic effect of the adsorbent. When used for extracorporeal circulation, the volume of the adsorbent layer 9 is 50 to 40011i! The degree is appropriate.

When the device of the present invention is used for extracorporeal circulation, there are two methods as follows: One method is to separate blood extracted from the body into plasma components and blood cell components using a centrifuge or membrane plasma separator, and then pass the blood t* through the device to purify it. One method is to return the blood to the body together with blood cell components, and the other method is to pass the blood taken out from the body directly through a nuclear device and purify it.

In addition, regarding the passage speed of blood or plasma, since the adsorption efficiency of the adsorbent is extremely high, the particle size of the adsorbent can be made coarser, and the degree of packing can be lowered, so the shape of the adsorbent layer can be changed. Regardless, high passing speeds can be provided. Therefore, a large amount of body fluid can be treated.

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

As described above, the adsorbent and adsorption device of the present invention adsorb and remove autoantibodies and/or immune complexes in body fluids at a high rate and specifically, are extremely compact, and are simple and safe. . It also has the advantage that sterilization operations can be carried out easily and reliably.

The present invention is applicable to the general use of purifying and regenerating body fluids such as autologous plasma, and is suitable for safe and reliable treatment of diseases related to the body's immune function, especially autoimmune diseases such as rheumatoid arthritis and systemic lupus erythematosus. Effective in treating diseases.

In addition, the adsorbent of the present invention can be used not only as a therapeutic device by filling it into a device, but also as an adsorbent for separating and purifying immunoglobulins, autoantibodies, immunoglobulin derivatives, and immunoglobulin complexes, and as a substrate for measuring these. It can also be used extremely effectively as a material.

Hereinafter, embodiments of the present invention will be explained in more detail with reference to Examples.

Reference example I CNBr activated Sepharose 4B (Sweden).

A hydrophobic compound containing various aromatic rings and hydrophobic amino acids or derivatives thereof was bonded to a compound (manufactured by Pharmacia, average particle size: 260 μm) by a conventional method, and excess active groups were blocked with ethanolamine. The amount retained is
The primary amino group of the remaining hydrophobic compound was converted to 4-phenylspiro[furan-2(3H), 1"-phthalane)-3,3"-
Reactively bonded with dione (“Fluram 0” manufactured by Roche),
It was calculated by measuring fluorescence at 475 to 490 nm (excitation wave 390 nm) and subtracting the amount of physical adsorption, which was separately experimented with unactivated Sepharose 4B.

The adsorbent after ethanolamine blocking was adjusted to pH 4-
After repeated washing with 0.1M acetate buffer and pH 8.5-0.1M boric acid buffer, the plate was washed with physiological saline, drained, and used for experiments.

In the adsorption experiment, 1 volume of human plasma and 1 volume of adsorbent were mixed, and 37
Incubate for 3 hours at °C. The amount of globulin and albumin after adsorption was measured using an A/G test kit (A/GB test manufactured by Wako, Wako Pure Chemical Industries, Ltd.).In addition, an adsorption experiment was conducted using unactivated Sepharose, which was used as a control. .

The results are shown in Table-1. It is clear from Table 1 that hydrophobic compounds containing at least one aromatic ring specifically and highly adsorb globulin.

Table 1 Furthermore, using the adsorbent (tryptamine/Sepharose 4B) IId after the adsorption experiment, the adsorbed globulins were quantified by immunoglobulin class.

After the adsorbent was washed five times with ice-cold PBS and dehydrated, the adsorbed globulin was eluted with ice-cold 50-O, 1M glycine-hydrochloride buffer (pH 2,5). The eluate was quantified by globulin class using single radial immunodefusion (SRID). For 5RID, a plate manufactured by ■Medical Biology Research Co., Ltd. was used. The detected amounts for each globulin class were 6 ■ for immunoglobulin G, 0.6 ■ for immunoglobulin M, and 1.4 ■ for immunoglobulin A. This shows that the adsorbent of the present invention adsorbs immunoglobulin with high activity regardless of its class.

Reference Example 2 Reference Example 1 except that cyclohexylamine and isoleucine methyl ester were used as hydrophobic compounds.
When we conducted an adsorption experiment in the same manner as above, we found that cyclohexylamine had a globulin adsorption rate of 12% and an albumin adsorption rate of 3%.
%, good results were obtained with isoleucine methyl ester, such as a globulin adsorption rate of 25% and an albumin adsorption rate of 6%.

Reference Example 3 An experiment similar to Reference Example 1 was conducted using a hydrophilic compound instead of a hydrophobic compound. The results are shown in Table-2. It is clear from this that hydrophilic compounds are significantly inferior in globulin adsorption ability and adsorption specificity.

Table 2 Example 1 Experiment similar to Reference Example 1 except that tryptophan, tryptophan methyl ester, tributatrine, and l-phenylalanine methyl ester were used as hydrophobic compounds and human rheumatoid arthritis patient plasma was used instead of human plasma. I did it. The ability to adsorb and remove rheumatoid factor (autoantibody) and immune complexes, which are malignant substances in rheumatoid arthritis, was measured.

Rheumatoid factor was measured using a latex agglutination test and a passive sensitization hemagglutination test.

The latex agglutination test is a detection method that measures the property of latex particles agglutinating when patient plasma containing rheumatoid factor is applied to polystyrene latex particles adsorbed with human γ-globulin. A dilution series is created and the rheumatoid factor concentration is evaluated at the plasma dilution rate at which latex particles no longer aggregate.

Plasma containing a high concentration of rheumatoid factor has a high dilution factor, while plasma with a low concentration has a low dilution factor.

Passive sensitization hemagglutination test was performed using rabbit-gamma on sheep red blood cells.
- Globulin is adsorbed, and the rest is the same as the latex agglutination test. Generally, the passive sensitization hemagglutination test is considered to be more specific for rheumatoid factor than the latex agglutination test.

A dilution series was prepared with glycine saline buffer, and the dilution ratio at which rheumatoid factor was negative in a latex agglutination test was determined. Latex agglutination tests were performed using a kit from Japan Freeze Drying Institute. Similarly, passive sensitization hemagglutination test)
Evaluation was made using [RAHA test, manufactured by Fuji Organ Pharmaceutical Co., Ltd.].

In addition, measurement of immune complexes can be performed using polyethylene glycol,
By complement hemolysis. In this method, immune complexes precipitated with polyethylene glycol are reacted with complement in the serum of a healthy human, and the amount of remaining complement is measured by the amount of hemolyzed red blood cells bound to antibodies. It evaluates quantity.

The operating method and conditions for this method are as follows.

(1) Pour 0.31 d of sample into a separation tube. 0.2MEDT
Add 50 μl of A and stir. Oxalate buffer (PB
S) Add 50μ and stir. Add 12.5% PEG (polyethylene glycol) (Mw 7500), stir, and let stand at 4° C. for 90 min.

(2) Centrifuge at 1700g 10+win at 4°C, and wash the resulting precipitate with 2.5% PEG1.0-. 1700
g Centrifuge for 15 sins and drain the supernatant.

(3) 37°C GVB” (gelatin veronal buffer containing divalent cations) 30 μl) Ic processing, dissolve the precipitate. Add 10 ui of pooled healthy human serum as a source of complement. 37°C 130C13O, immune complex react with complement.

(40,5X 10”/dEA (antibody-sensitized red blood cells) 1
0- is added and soaked in 60 ml of water at 37°C to promote hemolytic reaction due to residual complement.

(5) After the reaction, add 6.5 d of saline at 4°C, centrifuge, and measure the absorbance (OD, 4) of the supernatant.

(6) Calculate the inhibition rate of hemolysis relative to the control (serum from a healthy person), and set the unit to PEG-cc%.

Table 3 For EA, sensitized red blood cells (KW) for complement value measurement manufactured by Nippon Freeze Drying Research were used. In addition, globulin is reference example 1
It was measured by the method used in .

Unactivated Sepharose was used as a control in the same manner as in Reference Example 1, and the results are shown in Table 3 as removal rates when the control is set to 0.

From Table 3, it is clear that hydrophobic amino acids and their derivatives tryptophan, tryptophan methyl ester, tryptamine, and l-phenylalanine methyl ester specifically adsorb and remove globulin, rheumatoid factor, and immune complexes at a high rate. .

From the above table, it is clear that hydrophobic amino acids and their derivatives specifically and highly adsorb globulin, and furthermore, that they adsorb rheumatoid factors and immune complexes at a higher rate and specifically.

Comparative Example 1 An experiment similar to Example 1 was conducted using Protein A-Sepharose CL-4B (manufactured by Pharmacia, Sweden). Globulin was quantified using a high performance liquid chromatograph (2 columns 3000 SW, manufactured by Toyo Soda). In addition, in order to evaluate storage stability, Protein A-Sepharose CL-4B was suspended in physiological saline and
A similar experiment was conducted using a sample treated with 00''C for 160 minutes.

Sepharose CL-4B was used as a control.

The results are shown in Table-4.

Table 4 As is clear from the above table, protein A-Sepharose has a lower removal rate than hydrophobic amino acids and their derivatives, and when heat treated, its specificity decreases, making it undesirable.

Example 2 As a detection method, the property of chicken blood cells to aggregate when treated with patient plasma containing antibody A was measured. Otherwise, the amount of anti-DNA antibody is measured in the same manner as the latex agglutination test. The measurement was performed using a DNA test kit (Fuji Organ Pharmaceutical EL).
) was used.

Example 3 d, f tryptophan, d, f phenylalanine and d,
A l:1 oligopeptide of the oligomer was synthesized by the N-carboxylic anhydride method using n-hexylamine as an initiator. The ε-amino group of lysine was protected in advance with a carbobenzoxy group, and removed by a conventional method after oligopeptide synthesis. The degree of polymerization (number average) of the viable oligopeptide was determined by fluorescence analysis of the terminal amino group in the same manner as in Reference Example 1. Each oligomer was bound to activated CH-Sepharose 4B by a conventional method, blocked with glycine buffer and washed, and the same experiment as in Example 1 was conducted. In addition, the control was CH-5epharos blocked with ethanolamine.
It was set as e 4 B. The results are shown in Table-6.

The procedure was carried out in the same manner as in Example 1, except that systemic lupus erythematosus patient plasma was used instead of rheumatism patient plasma. The results are shown in Table-5.

Table 5 From the above table, hydrophobic amino acids and their derivatives are associated with immune complexes and anti-DN in plasma of patients with systemic lupus erythematosus.
It is clear that the A antibody is adsorbed more efficiently and specifically.

For the measurement of anti-DNA antibodies, formalin-fixed chicken blood cells were sensitized with DNA. It is clear that this specifically adsorbs and removes.

Reference Example 4 As a vinyl compound-based porous polymer, terpolymer copolymers consisting of hydroxylethyl methacrylate, ethylene glycol dimethacrylate, and glecidyl methacrylate had various average particle diameters and average pore diameters. An experiment was conducted using The epoxy density in the polymer is 1 mo1%
It is. l-tryptophan methyl ester was used as a hydrophobic amino acid derivative and fixed by a conventional method. Unreacted epoxide was blocked using ethanolamine. The amount retained was 4.3 μ/resin. As a control, a sample in which the entire amount of epoxide was blocked with ethanol agone was used. Similar to Reference Example 1, the sample was thoroughly washed and used for the experiment. The adsorbent 5rIdl was placed in a container as shown in FIG. 1 to prepare a globulin removal device.

A globulin adsorption experiment was conducted using the experimental system shown in FIG.

That is, a container 10 is filled with healthy human blood 11, pumped out at a flow rate of 1 H/min by a pump 12, sent to an immunoglobulin removal device 1, and then transferred to a drip chamber 1.
5 and a sampling port 13, and a tube 14 was arranged so that the sample was returned to the container 10. Note that 250 U of heparin was added to the blood.

After blood was circulated for 1 hour using the above device, the blood was sampled and the amount of globulin in the plasma was measured. The results are shown in Table-7.

In all cases, there was a small decrease in white blood cells and platelets before and after circulation.

Table 7 Reference Example 5 Using 0.5 denier Bembelb long fibers (manufactured by Asahi Kaei Kogyo), CNBr was activated by a conventional method to fix f-) liptophan methyl ester.

Blocking and washing were carried out in accordance with Reference Example 1. The amount retained is 9.
It was 8■/g Bemberg. An experiment similar to Example 1 was conducted using the adsorbent. The adsorbent was stored in the container shown in FIG.
．． 2g/ll1). Separately, untreated benbelub was used as a control for comparison. The results are shown in Table-8. Furthermore, there was a relatively small decrease in white blood cells and platelets before and after circulation.

Table 8 Example 4 Hydrophobic amino acids, 2-1 ml of liptophan and 1-phenylalanine, were immobilized on CNBr-activated Sepharose 4B (manufactured by Pharmacia, Sweden) by a conventional method to form an adsorbent. Blocking and washing were performed in the same manner as in Reference Example 1, and the amount retained was also measured in the same manner. The amount retained is
f-)Liptophan is 2.3■/d Cepharose (wet column volume, j!-Phenylalanine is 2.0■/d
It was Seph Arrows.

In the adsorption experiment, 1 volume of myasthenia gravis patient plasma was mixed with 1 volume of drained adsorbent, and the mixture was incubated at 37°C for 13 hours with shaking. Anti-acetylcholine receptor antibodies (autoantibodies) in the plasma after adsorption were measured by Lindstrom's two-antibody method. In addition, adsorption experiments were conducted using unactivated Sepharose 4B,
This was used as a control.

The results are shown in Table 9 in terms of removal rate when the control is set to 0.
It was shown to. From Table 9, it is clear that the adsorbent in which l-tryptophan and l-phenylalanine are immobilized on Sepharose 4B selectively and highly adsorbs anti-acetylcholine receptor antibodies, which are autoantibodies associated with myasthenia gravis. It is.

Table 9 It can be seen that globulin, rheumatoid factor, and immune complexes are selectively adsorbed and removed at a high rate.

Examples 5 to 9 and Comparative Examples 2 to 4 Tyrosine (Example 5), tryptophan (Example 6), phenylalanine (Example 7), histidine (Example 8), and valine (Example 9) were used as hydrophobic compounds. The same experiment as in Example 2 was conducted using DL-1ml leonine (Comparative Example 2), hydroxyproline (Comparative Example 3), and proline (Comparative Example 4) as hydrophilic compounds.

The results are shown in Table-10. From this result, it was found that an adsorbent bonded with a hydrophobic amino acid whose solubility in physiological saline at 25°C is 100 mmol/a or less,

[Brief explanation of drawings]

FIG. 1 is a cross-sectional view showing one example of the globulin compound adsorption apparatus of the present invention, and FIG. 2 is an explanatory diagram of a model experiment in an example. l...Autoantibody and/or immune complex removal device 2...Cylinder 3,3"...Filter 4.4
゛...Backing 5...Body fluid inlet 6...Cap 7...Body fluid outlet 8...Cap 9
... Adsorbent 10 ... Container 11 ... Blood 12 ... Pump 13 ... Sampling port 14 ... Tube 1
5... Drip chamber - Figure 2

Claims (1)

[Scope of Claims] 1. A hydrophobic amino acid or derivative thereof having a solubility in physiological saline at 25°C of 100 mmol/dl or less in an insoluble carrier, and/or an oligomer containing the hydrophobic amino acid or derivative thereof, which has a molecular weight An adsorbent for body fluid purification treatment of autoantibodies and/or immune complexes, characterized in that 0.1 to 30 mg of autoantibodies and/or immune complexes are bound to each ml of an insoluble carrier. 2. The adsorbent according to claim 1, wherein the hydrophobic amino acid or its derivative is tryptophan or its derivative. 3. Claim 1 in which the insoluble carrier is a hydrophilic carrier
The adsorbent according to any one of Items 1 to 2. 4. The adsorbent according to any one of claims 1 to 3, wherein the insoluble carrier is in the form of particles and has an average particle size in the range of 150μ to 2500μ. 5. The adsorbent according to claim 4, wherein the insoluble carrier is a porous polymer particle, and the average pore diameter is in the range of 300 Å to 9000 Å. 6. Claim 1 in which the insoluble carrier is fibrous and the fiber diameter is in the range of 0.02 denier to 10 denier.
The adsorbent according to any one of Items 1 to 3. 7. An oligomer containing an amino acid or its derivative and/or the compound having a solubility in physiological saline at 25° C. of 100 mmol/dl or less is bound to the insoluble carrier by a covalent bond, according to claim 1. adsorbent.