JPH0526509B2 - - Google Patents

Description

[Detailed description of the invention]

[Industrial Field of Application] The present invention relates to a blood treatment agent that specifically adsorbs endotoxins and globulins in blood. [Prior Art] Endotoxins are substances that exhibit pyrogenicity and toxicity when they enter the blood of mammals, and are heat-stable substances that cannot be decomposed even by high-pressure steam sterilization. For this reason, endotoxins must not be mixed into medical injection solutions or dialysate fluids for hemodialysis. Although it has been considered to use an ion exchange resin as an adsorbent for endotoxins, its adsorption capacity is small and it has not been put to practical use. In addition, normal human serum protein fractions are 5% α ₁ -globulin, 10% α ₂ -globulin, 10% β-globulin, 20% γ-globulin, and the remainder is albumin. This ratio increases or decreases depending on various diseases and is collectively called immunoglobulin disease. Selective adsorption and removal of these specific proteins in serum is
It is very difficult to do so without using expensive immunological techniques. Also, although the purpose is different, "Experimental and Applied Affinity Chromatography" (Ichiro Chibata, Tetsuya Tosa,
As an adsorbent for chromatography, hormones are immobilized on Sepharose beads (author: Yushi Matsuo, published by Kodansha Scientific). However, those using hydrophilic carriers such as Sepharose have a high degree of swelling in blood, resulting in poor blood permeability, making them unsuitable for blood treatment. It is also difficult to sterilize. On the other hand, if the particle size of Sepharose beads is increased, both the adsorption rate and adsorption capacity will be insufficient, making it unsuitable for practical use. [Problems to be Solved by the Invention] The present invention has the feature of being able to selectively adsorb and remove globulins and endotoxins in the blood, which can be used for clinical analysis and treatment. The present invention provides a blood treatment agent that can be applied to the treatment of bloodemia. [Means for Solving the Problems] A blood treatment agent made of a vinyl polymer having an aromatic nucleus into which a functional group represented by the following general formula (1) is introduced as a side chain. -X-Y (1) In the above formula, X represents a spacer group having a chain of 4 or more atoms, and within this spacer group, N,
It may contain O and S. Y represents a compound having a steroid skeleton. In the present invention, the vinyl polymer having an aromatic nucleus refers to a homopolymer of a vinyl monomer having an aromatic nucleus such as styrene, α-methylstyrene, vinyltoluene, etc. or a copolymer mainly composed of these. It is more preferable if these polymers are crosslinked. Further, it is more preferable that the polymer is reinforced with polyα-olefin, such as crystalline polypropylene or polyethylene, since mechanical properties will be improved. Examples include copolymers with polyvinyl compounds such as divinylbenzene or methylenebisacrylamide, as well as monovinyl compound polymer molded products crosslinked with formaldehyde, chlorosulfonic acid, and the like. Since crosslinked polymers have no fluidity and are difficult to mold, a method is preferably employed in which a molded polymer product is made into a fiber and then crosslinked. If the surface area of the fibers of the present invention is too small, the immobilization density will be low, but if the surface area is too large, the liquid permeability of the column packed with the fibers of the present invention will be poor.
0.01 or more and 50m ² /g or less, more preferably 0.05 or more
10m ² /g or less is preferable. In the above general formula (1), it is preferable that Y is bonded via a spacer having a chain of 4 or more atoms, rather than directly bonded to the backbone polymer, since the adsorption capacity is larger. Note that the spacer described herein refers to a molecule of arbitrary length that is introduced between the ligand and the carrier when immobilizing the ligand (a substance that has affinity with the target substance) on the carrier. The method of bonding Y to X used in the present invention is not particularly limited and may be any method that utilizes the condensation reactivity of the amino group, carboxyl group, or hydroxyl group of X. Specific examples of Y include colanic acids such as cholic acid, deoxycholic acid, chenodeoxycholic acid, and lithocholic acid, and steroid hormones such as testosterone, androsterone, estradiol, progesterone, and cortisone. In addition, the water content of the blood treatment agent of the present invention is preferably 0.7 to 3, more preferably 1 to 2; if it is too large, the liquid permeability will be poor and the pressure drop will be large, making it unsuitable for blood treatment. If it is too small, the swelling property will be poor. Practical adsorption capacity decreases. However, water content is the wet weight (W ₁ ) after the fibers are sufficiently swollen and then centrifugally dehydrated.
and dry weight (W _p ) using the following formula. Water content = (W ₁ /W _p )-1 If the density of the functional group represented by the general formula (1) is too low, the adsorption capacity will be low, so preferably 1 g of the polymer
It is present in an amount of 0.2 mmol or more, more preferably 1.0 mmol or more. Furthermore, when a functional group having a tertiary amino group is present as a side chain in the vinyl polymer having an aromatic nucleus of the present invention in addition to the functional group represented by the above general formula (1), the adsorption capacity is increased. It is preferable because it has the characteristic of increasing. Although there is no particular limitation on the chemical structure of such a functional group, a functional group represented by the following general formula (2) is usually preferably selected from the viewpoint of ease of production. In the above formula, R ₁ , R ₂ , R ₆ and R ₇ represent a hydrogen atom or a lower alkyl group. Even in the case of a group in which such a functional group coexists, the general formula (1)
The functional group is preferably present in an amount of at least 0.01 mmol per gram of the polymer. When the functional groups represented by general formula (2) are present together, the sum of the functional groups represented by general formulas (1) and (2) is at least 1.0 per gram of the polymer.
The amount is selected from the viewpoint of adsorption capacity, preferably 2.0 mmol or more. An example of the production of the blood treatment agent of the present invention is as follows:
A molded article made of an α-haloacetamidomethylated aromatic vinyl polymer (Japanese Patent Application Laid-Open No. 12008/1983) is mixed with a solution of an amine represented by the general formula (3), or the amine and an amine represented by the general formula (4). First, a carrier is obtained by immersing it in a mixed solution. R ₆ -NH-R ₇ (4) Next, the basic skeleton of the following general 5β shown by equation (5)
The blood treatment agent of the present invention can be produced by immersing the carrier in a mixed solution of -colanic acid and N,N'-dicyclohexylcarbodiimide. The blood treatment agent of the present invention can be used by filling it into a column for extracorporeal circulation and bringing it into continuous or intermittent contact with whole blood or plasma, or by filling it into a syringe and placing it inside the syringe. Methods include, but are not limited to, methods of aspirating whole blood or plasma and then returning the whole blood or plasma to the body. [Example] Example 1 Polypropylene (Mitsui “Noblen” J3HG)
50 parts as an island component, polystyrene (“Styron”)
666) 46 parts, polypropylene (Sumitomo “Noblen”)
Sea-island composite fiber containing 4 parts of WF-727-F) as a sea component (number of islands: 16, monofilament fineness: 2.6 denier,
Tensile strength 2.9g/d, elongation 50%, number of filaments
42) 100g, N-methylol-α-chloroacetamide 120g, nitrobenzene 800g, 98% sulfuric acid
It was immersed in a mixed solution consisting of 800 g and 1.7 g of paraformaldehyde, and reacted at 20°C for 1 hour. The fibers were taken out from the reaction solution, placed in ice water at 0°C to stop the reaction, and then washed with water.
Nitrobenzene adhering to the fibers was extracted and removed with methanol. This fiber (fiber A) was vacuum dried at 50°C to form a chloroacetamide methylated fiber.
Obtained 140g. Dodecamethylene diamine 23.7g 50% of 2100ml
68 g of the fiber A obtained above was added to the mixed solution obtained by dissolving it in a dimethylamine aqueous solution, and the mixture was heated to 48 g at room temperature.
Allowed time to react. The fibers were taken out and thoroughly washed with dilute hydrochloric acid and water to obtain mixed aminated fibers (fiber C). The amount of dimethylamino groups in this fiber C is 2.51
Milliequivalent/g, amount of dodecamethylenediamine is 0.18
It was milliequivalent/g. 1N-30g of fiber C
After treatment with an aqueous NaOH solution, the fibers were thoroughly washed with water and then dried to obtain an aminated fiber (fiber D). Next, add 3.25 g of deoxycholic acid to 200 ml of N,
It was dissolved in N dimethylformamide (hereinafter referred to as DMF) (liquid E). On the other hand, 2.05 g of N,N'-dicyclohexylcarbodiimide (DCCD) was dissolved in 20 ml of DMF and added to the above solution E. Immediately stir to make a homogeneous solution and add 6g of fiber D (dry weight) into it.
was added and allowed to react at room temperature for 72 hours. The reaction fibers were taken out, packed into a chromatography column, and washed with 250 ml of DMF, washed with a 1N-NaOH aqueous solution, and further washed with water to obtain fibers (1), which are the blood treatment agent of the present invention.
This product had an exchange capacity of 2.46 meq/g and a Cl type water content of 0.87. The difference between this exchange capacity and the exchange capacity of fiber C was regarded as the amount of deoxycholic acid immobilized on the fiber. The immobilized amount was 58 mg/g. Example 2 The following adsorption experiment was conducted on the fiber (1) of the blood treatment agent of the present invention obtained in Example 1. Add 200mg of the above fiber (1) to 10ml of human serum and heat at 37°C.
After shaking for 3 hours, the amount of blood components in the supernatant was determined, and the adsorption rate for each component was determined. The results are shown in Table 1.

[Table] In the table, comparative sample 1 was prepared by soaking 100 g of fiber A in 10% aqueous ethanol 2 containing 100 g of potassium iodide, and
5 g of this fiber was immersed in a solution consisting of 30 g of tri-n-propylamine, 60 g of dimethyl sulfoxide, and 60 g of ethanol, heated at 55° C. for 12 hours, and diluted with dilute hydrochloric acid. and tri-n-propylammonium chloride acetamidomethylated fibers obtained by washing with 1M saline. The neutral salt decomposition capacity of this product was 1.64 milliequivalents/g, there was no weak basic group content, and the Cl type water content was 1.57. Table 1 shows that the fiber that is the blood treatment agent of the present invention adsorbs globulin and HDL-cholesterol well. Example 3 The following endotoxin adsorption experiment was conducted on the fibers of the blood treatment agent of the present invention obtained in Example 1. Prepare 5 test tubes (outer diameter 24 mm) for each sample, and hold 20 mg, 40 mg, 60 mg, 100 mg, and 200 mg in each tube.
of fiber, then add 10 ml of lipopolysaccharide aqueous solution (E.
Coli055: B5, trichloroacetic acid extraction method, 0.11 mg/ml manufactured by Deif Collaborative Laboratories) was added, and the temperature was 37°C.
After shaking for 4 hours, it was filtered using a No. 2 qualitative filter paper, and the concentration of lipopolysaccharide in the filtrate was determined using the phenol/sulfuric acid method (2 ml of sample + 1 ml of 5% phenol water + 1 ml of 5% phenol water +
Measured with 5 ml of 98% sulfuric acid: 485 mμ). The difference between the lipopolysaccharide concentration in the mother liquor and the initial concentration (0.11 mg/ml) was regarded as the amount of lipopolysaccharide adsorbed, and an isothermal adsorption line was determined. In order to compare the adsorption capacity of each fiber, the adsorption capacity when the mother liquor concentration was 0.05 mg/ml was determined from the isothermal adsorption curve, and Table 2 shows the results.

[Table] In the table Comparative sample (1) is the same tri-n-propylammonium chloride acetamidomethylated fiber used in Example 2. Comparative sample (2) is the result of using an ion exchange resin (manufactured by Organo Co., Ltd.: Amberlite IRA-938). Table 2 shows that the fibers of the blood treatment agent of the present invention have excellent adsorption properties for endotoxins. [Effects of the Invention] The present invention has an excellent ability to selectively adsorb globulins and endotoxins in the blood, and at the same time can achieve its objectives with a high fluid passage rate and adsorption capacity. The present invention enables miniaturization of the device at a practical level, and is a blood processing agent convenient for clinical analysis and treatment.

Claims (1)

[Scope of Claims] 1. A blood treatment agent comprising a vinyl polymer having an aromatic nucleus into which a functional group represented by the following general formula (1) is introduced as a side chain. -X-Y (1) In the above formula, X represents a spacer group having a chain of 4 or more atoms, and within this spacer group, N,
It may contain O and S. Y represents a compound having a steroid skeleton.