KR100470502B1 - Adsorbent carrier used in direct hemoperfusion and method for reducing the particle size thereof - Google Patents

Abstract

The present invention relates to a sulfated polysaccharide having an intrinsic viscosity of 0.005 to 0.5 dl / g and a sulfur content of 5 to 22 wt%, and / or a salt thereof in an amount of 0.02 to 200 mg per ml of water-insoluble carrier, A particle hardening method of a carrier for an adsorbent for use in direct blood perfusion. The method of the present invention enables the particle hardening of the adsorbent without causing pain and pain.

Description

Adsorbent carrier used for direct blood perfusion and its small size hardening method {ADSORBENT CARRIER USED IN DIRECT HEMOPERFUSION AND METHOD FOR REDUCING THE PARTICLE SIZE THEREOF}

The present invention relates to a carrier for an adsorbent for use in direct blood perfusion and to a particle hardening method thereof.

In order to adsorb | suck and remove the malignant substance in blood, it is known to use the adsorbent which immobilized the ligand to the carrier. The adsorption removal method includes a method of perfusion of blood and continuous processing. The perfusion method separates blood into blood cells (blood cell components) and plasma components to perform adsorption removal treatment on only plasma components, and blood cells and plasma. There is a direct blood perfusion method in which the components are adsorbed and removed in the form of whole blood without separating the components.

In general, in order to increase the adsorption efficiency, the effective surface area may be increased by reducing the particle diameter of the adsorbent and its support. This method is easy for direct blood perfusion by, for example, batch processing or adsorption treatment of only plasma components having low viscosity and not containing blood cells.

On the other hand, the direct blood perfusion method for blood is generally adopted by filling an adsorbent in a column or the like and passing the whole blood therein (also called an online method). After admixing with blood and adsorbing, the adsorbent is filtered to return whole blood to the patient (batch). Either way, the whole blood is in direct contact with the adsorbent, so that adhesion, breakage, activation of blood cells, such as platelets, or activation of the coagulation system occurs and the blood proceeds to coagulate. This phenomenon is greatly influenced by the material and the particle diameter constituting the carrier for the adsorbent, and in particular, the smaller the particle diameter, the more remarkable. Therefore, even if an adsorbent with a smaller particle size is used, the adsorption efficiency is increased, but the particle size hardening of the adsorbent is difficult in the direct blood perfusion method from the viewpoint of ensuring stable blood flow. In particular, in the online method, when blood cell adhesion or aggregation occurs, a blood flow path is not secured, thereby causing a problem in blood flow. In view of this, an adsorbent carrier having an average particle diameter of more than about 400 mu m is currently used.

In the batch method, the blood flow is not particularly a problem, and a small particle size adsorbent may be used as compared to the online method. However, even in this case, a small particle size causes adhesion of the blood cells and coagulation of the blood cells, and thus, there is a limit to small particle hardening.

Thus, the adsorbents used for direct blood perfusion for whole blood are not necessarily satisfied with only small particle diameters, and it is important to suppress the adhesion of blood cells and the like as much as possible.

An object of the present invention is to small-size the carrier for direct blood perfusion while maintaining excellent blood flow.

The present inventors have made intensive studies on heparin similarity, toxicity, interaction of blood cells and proteins of sulfated polysaccharides, and, when carrying out carrier processing with sulfated polysaccharides or salts thereof, even if the carrier for direct blood perfusion is further reduced in size, The present invention has been completed by discovering that high blood cell permeability is exhibited, activation is also inhibited, and blood coagulation time is not shortened.

Disclosure of the Invention

That is, the present invention relates to a carrier for an adsorbent for use in direct blood perfusion, wherein the sulfated polysaccharide and / or salt thereof is bound to a water-insoluble carrier, and a particle hardening method thereof.

The sulfated polysaccharide or salt thereof used in the present invention does not activate blood cells, does not cause pain in blood coagulation, etc., and release factors released from blood cells by other factors (bioactive amines such as serotonin and histamine) Overreaction with basic polypeptides such as platelet factor 4 (PF 4), β-thromboglobulin (βTG) and proteins having affinity with heparin such as thrombospondin, fibronectin, and neutral proteases such as granulocyte elastase Has a function of adsorbing and trapping, and is also useful as a ligand.

The sulfated polysaccharide or its salt bound to the water-insoluble carrier is preferably an intrinsic viscosity of 0.005 dl / g or more and 0.5 dl / g or less from the viewpoint of inhibiting adhesion of blood cells, and is 0.007 dl / g or more and 0.4 dl / g or less. In this regard, the immobilization can be more preferably achieved, and more preferably 0.008 dl / g or more and 0.2 dl / g or less, and is preferable in view of suppressing nonspecific adsorption of plasma components by adding such advantages.

In addition, the sulfur content of the sulfated polysaccharide or its salt is preferably 5 wt% or more and 22 wt% or less in terms of inhibiting adhesion of blood cells, and more preferably 8 wt% or more and 22 wt% or less in terms of anticoagulability, In particular, 13 wt% or more and 22 wt% or less are preferable in terms of more stable inhibition of adhesion of blood cells and expression of anticoagulation.

In addition, the amount of the sulfated polysaccharide or its salt bound to the water-insoluble carrier is preferably 0.02 mg or more and 200 mg or less per ml of the water-insoluble carrier, since it suppresses adhesion of blood cells and anticoagulant, and is 0.1 mg or more and 100 mg or less. It is more preferable at the point which suppresses excessive anticoagulability, and it is especially preferable that it is 0.5 mg or more and 40 mg or less in terms of practical surface use, the outstanding inhibitory effect of blood cell cell, and an anticoagulant expression.

And the intrinsic viscosity is 0.005 dl / g or more and 0.5 dl / g or less, and the sulfuric acid polysaccharide having a sulfur content of 5 wt% or more and 22 wt% or less and 0.02 mg or more and 200 mg or less per 1 ml of a water-insoluble carrier. Binding in amounts is desirable in ensuring blood flow.

Representative examples of sulfated polysaccharides or salts thereof suitable for use in the present invention include, for example, heparin, dextran sulfate, chondroitin sulfate, chondroitin polysulfuric acid, heparan sulfate, keratan sulfate, heparin sulfate, xylan sulfate, Ronin Sulfate, Chitin Sulfate, Chitosan Sulfate, Cellulose Sulfate, Agarose Sulfate, Agalopectin Sulfate, Pectin Sulfate, Inulin Sulphate, Alginate Sulfate, Glycogen Sulfate, Polylactose Sulfate, Carrageenan Sulfate, Starch Sulfate, Polyglucose Sulfate, Lami Sulfated polysaccharides, such as a flylin sulfuric acid, a galactan sulfuric acid, levane sulfuric acid, and mefesulfate, these potassium salts, a sodium salt, etc. are mentioned. This invention is not limited only to these. Examples of the most preferable examples in terms of production cost and raw material acquisition include heparin, dextran sulfate, and chondroitin polysulfuric acid.

In the present invention, the carrier capable of small particle hardening may be water insoluble, and may be a soft gel or a hard gel. As a soft gel, dextran, agarose, polyacrylamide, etc. are mentioned, for example. However, since it is preferable that it is pressure-resistant (small compaction) in the point used for direct blood perfusion, a hard gel piece is preferable. Among the hard gels, polymer hard gels are preferred because the porous body can be obtained relatively easily and the water swellability is small. The polymer hard gel in the present invention includes polymethyl methacrylate, polyvinyl alcohol, styrene-divinylbenzene copolymer, crosslinked polyvinyl alcohol, crosslinked polyacrylate, crosslinked vinyl ether-maleic anhydride copolymer, crosslinked styrene- Copolysaccharides, synthetic polymers, etc. are coated on the surfaces of synthetic polymers such as maleic anhydride copolymer and cross-linked polyamide, and inorganic porous materials such as porous polymers made of natural polymer compounds such as cellulose, porous glass, and porous silica gel, and surfaces thereof. One thing is included. These polymer hard gels may be used alone or in combination of two or more thereof. However, the present invention is not limited only to these.

Hard gel and soft gel can be distinguished by the following method. That is, the gel is uniformly filled in a glass cylindrical column (inner diameter 9 mm, column length 150 mm) equipped with a filter having a pore diameter of 15 μm at both ends, and after flowing an aqueous liquid, the relationship between pressure loss and flow rate is hard gel. In the case of a nearly straight line, the soft gel deforms and consolidates when the pressure exceeds a certain point, so that the flow rate does not increase. In the present invention, when the above-described column is used, those having the above linear relationship up to 0.3 kg / cm 2 or more are referred to as hard gels.

The shape of the carrier may be granular or somewhat deformed. According to the method of the present invention, the carrier can be considerably small (particle size of about 30 mu m). In the batch system, in order to reliably separate blood and adsorbent after the adsorption treatment, it is preferable to have an average particle diameter of about 30 µm or more. When used in the online method of direct blood perfusion, direct blood perfusion may become difficult due to individual differences when the lower limit average particle diameter is less than 80 µm. The upper limit is not particularly limited, but the average particle diameter is preferably 5000 µm or less in adsorption performance. It is more preferable that the thickness is 100 µm or more and 1000 µm or less, and particularly preferably 120 µm or more and 800 µm or less in terms of the performance and adsorption performance of stable direct blood perfusion in the on-line system. In addition, in the sense of stably performing direct blood perfusion and inadvertently activating blood cell cells to release the release factor, it is the gist of the present invention to limit the average particle size distribution of the particles. If the average particle size distribution is too wide, it causes the adhesion or activation of blood cells, and if the distribution is narrow, the physical adhesion or activation due to turbulence or the like shows a tendency to suppress. Therefore, in the present invention, it is preferable that the particle size of 80% by volume or more of the carrier particles is distributed within ± 75% of the average particle diameter, and the activation of the blood cell by the adsorption device itself is suppressed, and the release factor of the blood cell is In order to adsorb | suck and remove efficiently, it is more preferable to distribute within ± 50%. Especially preferably, it is distributed within +/- 25%.

The method of the present invention is to bind the sulfated polysaccharide and / or salt thereof to the water insoluble carrier. In addition to the conventionally known coating method, there are immobilization methods such as physical adsorption method, ion bonding method, and covalent bonding method, but are not particularly limited thereto. Sulfated polysaccharides are safer than synthetic polymer materials having various antithrombogenic properties, which are currently being actively researched and developed. However, since they are undesirable to enter the body, a method of covalent bonds that do not drop well is more preferable. . It is also important that the sulfated polysaccharides do not desorb during sterilization. Therefore, a firm covalent bonding method is also preferable in this regard, and even if an ionic bonding method is used, it is preferable to covalently crosslink the sulfated polysaccharides. desirable. In particular, by introducing a spacer between the carrier and the sulfated polysaccharide, it is possible to suppress interaction with proteins and blood cells.

In the immobilization of the sulfated polysaccharide, it is preferable that a functional group capable of easily reacting with the sulfated polysaccharide is present on the surface of the carrier. Representative examples of such functional groups include amino groups, carboxyl groups, hydroxyl groups, thiol groups, acid anhydride groups, succinylimide groups, chlorine atoms, aldehyde groups, amide groups, epoxy groups and the like. These functional groups are introduce | transduced into a carrier by a well-known method as needed. Specific examples of the immobilization by the covalent bonding method include cyanide halide, epichlorohydrin, bisepoxide, and triazine halide. For example, the bonding is particularly strong and the risk of desorption of sulfated polysaccharides is increased. Less epichlorohydrin method is most preferred.

The method of the present invention utilizes the action of inhibiting adhesion of blood cells of sulfated polysaccharides or salts thereof and extending blood coagulation time. Therefore, by immobilizing a ligand for a specific purpose on a carrier hardened by the method of the present invention, it is possible to efficiently adsorb and remove specific blood substances while maintaining good blood flow. A conventionally well-known method can be used for fixation of a ligand. Such ligands include (1) a substance having affinity for a specific substance derived from a living body, for example, a fragment such as an immunoglobulin or an immunoglobulin Fab, Fab ', F (ab') ₂ , or a glucosaminoglucan such as heparin; Sugars and the like; (2) Substances exhibiting affinity through electrostatic interaction such as (poly) anions or (poly) cations such as carboxyl groups, sulfonic acid groups, substances having anionic groups such as sulfuric acid groups or phosphoric acid groups, or amino groups or substituted amino groups Materials having cationic groups; (3) substances exhibiting affinity by hydrophobic interactions; For example, the substance which has a long chain alkyl group, an aromatic group, a silicon compound, a fluorine compound, etc. is not limited to these.

The adsorbent using the carrier of the present invention can be used in any of the above-described batch method and online method, but is not limited thereto, and direct blood perfusion method of returning the whole blood to the patient's body after the adsorption treatment. Can be applied in any way.

Further, as described above, the sulfated polysaccharide or salt thereof used in the present invention, a physiologically active amine such as a release factor released from blood cells, such as serotonin and histamine; Basic polypeptides such as platelet factor 4, β-thromboglobulin, platelet-derived growth factor (PDGF), and basic fibroblast growth factor (bFGF); Proteins having affinity with heparin such as thrombospondin, fibronectin, epidermal growth factor (EGF), etc .; Factors causing an overreaction to living bodies such as neutral proteases such as granulocyte elastase; It has the effect of adsorbing and capturing lipoproteins such as LDL and VLDL in the blood. Therefore, the carrier for the adsorbent obtained by the present invention, even without immobilizing other ligands, is useful as an adsorbent for adsorbing and removing the release factor or lipoproteins as direct blood perfusion alone.

Hereinafter, although the method of this invention is demonstrated concretely based on an Example, this invention is not limited only to this Example.

In the following examples, the interaction of the small-size hardening carrier and the blood cell prepared according to the method of the present invention and the variation of blood coagulation time were examined, and the blood flow evaluation in direct blood perfusion was evaluated. In general, blood is caused by the rapid attachment of blood cells to the foreign body surface or activation of the coagulation system when the blood comes into contact with a foreign substance that does not exist in the blood vessel. Therefore, using these items as an index evaluates the interaction between the carrier and the blood. Suitable for In Examples, the results of the interaction with blood cells are evaluated by the passage rate (%) and the non-adhesion rate (%), and in all cases, the upper limit is 100% and the higher the value, the more the interaction between the various carriers and the blood cells. Means weak action. In addition, regarding the blood coagulation time, two types of coagulation time were measured: an endogenous coagulation time, which is likely to reflect the influence of foreign substance contact, and an exogenous coagulation time activated by a tissue factor or the like. The blood coagulation time is preferably invariant or prolonged (from the viewpoint of suppressing coagulation due to foreign substance contact) rather than being shortened by activation by contact with a carrier. Also in this embodiment, when it is short compared with the numerical value at the time of blood collection, it shows that blood becomes easy to coagulate and it can be judged that compatibility with a carrier is low. On the other hand, in the case of invariant or prolonged tendency, it can be judged that the suitability is high in the sense that the blood is not easily coagulated.

Example 1

20 ml of reverse osmosis water (RO water, Yamato Pure Line RO 21) was divided into 20 ml of porous cellulose gel (manufactured by Chisso Co., Ltd., with an average particle diameter of 80 µm and particles with a volume of 80 vol% or more within an average particle diameter of ± 10%) by 20 ml. And Yamato Chemical Co., Ltd.) are added, and internal temperature is heated up to 40 degreeC. 11 ml of 2M NaOH was added here, and it stirred for 30 minutes at 40 degreeC. Next, 3.6 ml of epichlorohydrin is added and reacted at 40 degreeC for 2 hours. After completion of the reaction, the gel is washed with about 2 L of RO water to obtain an epoxidized gel. The amount of epoxy groups introduced is 13 μmol / g.

15 ml of the epoxidized gel was divided into precipitates, and 10.5 g of dextran sulfate (molecular weight: about 2,000, intrinsic viscosity: 0.012 dl / g, sulfur content: 13 wt%) was added to a solution obtained by dissolving in 19 ml of reverse osmosis water. Stir. Subsequently, after adjusting to pH10 with 2N NaOH, it is made to react at 45 degreeC for 24 hours, and dextran sulfate is immobilized.

RO water is added to 115 µl of monoethanolamine to make the whole amount 3.3 ml. An aqueous solution is added to the dextran sulfate fixed gel (sedimentation volume 15 ml), followed by 23 ml of RO water. It is made to react at 45 degreeC for 2 hours, ring-opening reaction (sealing reaction) of an unreacted epoxy group, and an adsorbent (henceforth CKA80-DS) is obtained. The dextran sulfate fixed amount of this carrier is 2.09 mg / mL (gel sedimentation volume).

By the same method, the carrier for an adsorbent is produced using the water-insoluble carrier shown in Table 1.

And manufacture number 5 is produced by the following method.

Epoxidized styrene-divinylbenzene copolymer (POROS 50 EP manufactured by Persecution Biosystems Co., Ltd., average particle diameter of 36 μm, particles of 80% by volume or more are referred to as the average particle diameter of within ± 20%, hereinafter referred to as POR-EP) Weigh 10 ml by sedimentation volume. After completely dissolving 7.0 g of dextran sulfate and 5.0 ml of reverse osmosis water as used in CKA80-DS, the mixture was mixed with the weighed POR-EP and adjusted to pH 9.2 by adding 2 M NaOH at room temperature. Subsequently, the mixture is stirred at 45 ° C. for 24 hours in a constant temperature water bath to fix dextran sulfate. Subsequently, reverse osmosis water is added to 77 µl of monoethanolamine to make the whole amount 2.2 ml. This solution is added to 10 ml (precipitated volume) of the dextran sulfate-fixed gel, and then 15.3 ml of reverse osmosis water is added and reacted at 45 ° C for 2 hours to encapsulate unreacted epoxy.

Manufacturing number Abbreviation Water Insoluble Carrier material Average particle size (㎛) Particle size distribution ¹⁾ (%) Immobilization Amount ²⁾ (mg / ml) 1 2 3 4 5 CKA80-DSCKA200-DSCKA250-DSCKA350-DSPOR-DS Porous Cellulose Gel Porous Cellulose Gel Porous Cellulose Gel Porous Cellulose Gel Epoxylated Styrene-Divinylbenzene Copolymer 8020025035036 ± 10 ± 10 ± 10 ± 20 ± 20 2.092.602.272.142.26 1): indicates what percentage of the average particle diameter is included in the particle size of particles of 80% by volume or more 2): based on 1 ml of gel sedimentation volume

Example 2

Adsorption Experiment

The dextran sulfuric acid immobilized porous cellulose gel prepared in Examples 1 to 5 was washed with heparin physiological saline (adjusted to have a final concentration of heparin of 7 U / ml) to equilibrate heparin. After defoaming the gel, 1 ml of the gel is packed into a polypropylene minicolumn (inner diameter of 9 mm, height of 16 mm: manufactured by Termo Co., Ltd.). A polycolumn tube (inner diameter 1 mm, outer diameter 3 mm, length 1 m) was mounted on the column inlet side to produce a minicolumn with a blood circuit.

Blood is carefully drawn using heparin as an anticoagulant and using an 18 G needle from a healthy common person. The collected blood is then placed in a Teflon Erlenmeyer flask (100 mL content, manufactured by Shanwa Co., Ltd.), rotated at low speed in a stirring bar in a 37 ° C incubator to start blood flow into the column at a flow rate of 0.5 mL / min.

The time when blood flowed out of the minicolumn outlet side was the start of blood flow. After that, a predetermined amount of blood was extracted from the column outlet side every 5 minutes, and the number of blood cells (red blood cells, white blood cells, platelets) in the blood was counted with a blood cell counter (Microcell). Counter) Measured with CC-180, manufactured by Sysmex Co., Ltd., the pass rate is calculated according to the following equation. The results (three times average) are shown in Table 2. And all the red blood cells passed through were 100%.

Passage rate (%) = (column cell on the outlet side / blood cell count on blood collection) × 100

For comparison, the blood flowability is examined in the same manner for the porous cellulose gel without immobilizing dextran sulfate. The results are shown in Table 2 together. Those having an average particle diameter of 80 µm, 200 µm, 250 µm and 350 µm are abbreviated as CKA80, CKA200, CKA250 and CKA350, respectively. Also in these carriers, the passage rate of red blood cells was 100%.

Pass rate (%) CKA80-DS CKA80 CKA200-DS CKA200 CKA250-DS CKA250 CKA350-DS CKA350 Leukocytes 10 minutes after blood flow 20 minutes after blood flow 30 minutes after blood flow 968685 756763 979892 868075 1009990 927975 10010097 948380 Platelet 10 minutes after the blood 20 minutes after the blood 30 minutes after the blood 487378 383678 657592 514777 588290 474284 598391 534976

As can be seen in Table 2, the passage rate of leukocytes indicates that the CKA350-DS, CKA250-DS, and CKA200-DS all pass close to 100%. In addition, CKA80-DS shows a passage rate of about 90%. The platelet transit rate was almost no difference in CKA200-DS, CKA250-DS, and CKA350-DS, and it was about 50% at the beginning of the onset of blood flow, but increased over time, and reached 80% after 20 minutes at the time of onset of blood flow. In addition, CKA80-DS showed about 40% at the beginning of a blood flow, and after that time, it showed the passage rate increase, and the 30-minute numerical value of completion | finish of evaluation was about 75%.

In the case of the gel without immobilization of dextran sulfate, the passage rate of leukocytes was 10% to 20% lower than that of the gels corresponding to the particle sizes of CKA80, CKA200, CKA250, and CKA350 immobilized with dextran sulfate. On the other hand, the particle size of each of the dextran sulfate-free gels in the platelet passage rate was 10% to 40% lower than that of the dextran sulfate-fixed gel.

Example 3

After using the experimental system prepared in Example 2 and passing a minicolumn packed with dextran sulfate-fixed gel of various particle diameters prepared in Example 1 of the same method as the same example, blood coagulation time was measured. Measure with apparatus (Blood Coagulation analyzer CA-100 Sysmex Co., Ltd.). Blood coagulation time (exogenous and endogenous) was measured by collecting a predetermined amount of blood on the column exit side every 10 minutes from the beginning of blood flow, centrifugation at 3000 rpm for 15 minutes, and neutralizing heparin in plasma with protamine sulfate. do. It is also measured for gels of each particle diameter unfixed dextran sulfate for comparison. The results (three times average) are shown in Table 3.

CKA80-DS CKA80 CKA200-DS CKA200 CKA250-DS CKA250 CKA350-DS CKA350 Exogenous coagulation time (sec) (at the time of blood collection) 10 minutes after blood flow 20 minutes after blood flow 30 minutes after blood flow (20) 542724 (20) 201618 (20) 252120 (20) 201919 (20) 252323 (20) 232324 (20) 242122 (20) 242323 Endogenous coagulation time (seconds) (at the time of blood collection) 10 minutes after blood flow 20 minutes after blood flow 30 minutes after blood flow (82) 290516881016 (82) 768268 (82) 1393459368 (82) 767275 (82) 588433278 (82) 969895 (82) 396280268 (82) 1009695

Exogenous coagulation time (prothrombin time) did not change in blood collection values in CKA350-DS and CKA250-DS, but CKA200-DS tended to extend slightly after 10 minutes from the start of blood flow treatment. In CKA80-DS, 10 minutes after commencement of blood flow treatment, the value was about three times higher than blood collection. Thereafter, the extrinsic coagulation time shows a tendency to recover rapidly, and at the end of blood transfusion, there is almost no difference with gels of other particle diameters.

With regard to the endogenous solidification time (activation partial thromboplastin time), it was confirmed that all gels of any particle size were significantly extended at the evaluation time of 10 minutes and 20 minutes. As time passed, the coagulation time tended to be close to the value at the time of blood collection, but did not recover 30 minutes after the end of the evaluation.

On the other hand, in the dextran sulfate unfixed gel, the gel of any particle size and the blood coagulation time (both exogenous and endogenous) retained the values at the time of blood collection and showed little variation.

Example 4

POR-DS prepared in Preparation No. 5 of Example 1 was washed with 20 times the amount of physiological saline solution (prepared to have a final heparin concentration of 7 U / ml) of the gel sedimentation volume.

POR-DS was placed in a polypropylene test tube with 0.5 mL of sedimentation volume (800 rpm, gel sedimentation was performed for 30 seconds), and then 3 mL of blood (7 U / mL) anticoagulated with heparin by removing the supernatant. After each addition, the lid was sealed, and shaken well for 30 minutes at 37 ° C. at 60 times / min.

30 minutes after the start of shaking, by measuring the number of platelets present in the blood, the ratio of the number of platelets not adhered to the gel (specific adhesion ratio) is calculated by the following equation. The sample was centrifuged at 3000 rpm for 15 minutes, and 0.5 ml of supernatant was collected to neutralize heparin with protamine sulfate, and the blood coagulation time (endogenous and exogenous) was measured in the same manner as in Example 3. In addition, the result (mean twice) of the raw material POR-EP which did not fix the dextran sulfate was shown in Table 4 for comparison.

Specific adhesion rate (%) = (platelet count in sample gel-added blood / platelet count in gel unadded blood) * 100

Non-Adhesion Rate (%) Exogenous clotting time (seconds) Endogenous solidification time (seconds) POR-EPPOR-DS during blood collection -5770 182158 90841945

As can be seen from Table 4, the specific adhesion rate of platelets to POR-DS shows a high value of 70%. In terms of blood coagulation time, the coagulation time of the exogenous system was extended by about three times the value at the time of blood collection, and the endogenous coagulation time was extended by about 20 times the value at the time of blood collection.

On the other hand, in POR-EP without immobilizing dextran sulfate, the ratio (non-adhesion rate) of platelets that did not adhere to the gel shows a low value of about 57% compared to the control without adding gel. On the other hand, the blood coagulation time maintained the values at the time of blood collection for both the coagulation time of the exogenous and the endogenous, and no large variation was recognized.

By using the method of the present invention, adhesion of leukocytes or platelets can be greatly suppressed, and blood coagulation time can be significantly extended, and as a result, the particle size of the water-insoluble carrier used for direct blood perfusion can be further reduced in size. As a result, it is possible to simultaneously achieve an increase in adsorption capacity and an improvement in the permeability of direct blood perfusion.

Claims (8)

(Iii) sulfated polysaccharides, salts thereof, or sulfated polysaccharides and salts thereof, and (ii) these sulfated polysaccharides, salts thereof, or sulfated polysaccharides and ligands different from the salts that adsorb a particular substance during direct blood perfusion treatment. Adsorbent used for direct blood perfusion formed by binding to a water-insoluble carrier. (Iv) sulfated polysaccharides, salts thereof, or sulfated polysaccharides and salts thereof having an intrinsic viscosity of not less than 0.005 dl / g and not more than 0.5 dl / g of sulfur content of not less than 5% and not more than 22% by weight and (ii) direct blood perfusion An adsorbent for use in direct blood perfusion in which a sulfated polysaccharide, a salt thereof, or a sulfated polysaccharide and a ligand different from the salt thereof is bound to a water-insoluble carrier during the treatment. The adsorbent according to claim 1 or 2, wherein the sulfated polysaccharide, its salt, or sulfated polysaccharide and its salt are bonded to the water insoluble carrier in an amount of 0.02 mg or more and 200 mg or less per ml of the water insoluble carrier. The adsorbent according to claim 1 or 2, wherein the water-insoluble carrier has an average particle diameter of 30 µm or more. (I) sulfated polysaccharides, salts thereof, or sulfated polysaccharides and salts thereof; and (ii) these sulfated polysaccharides, salts thereof, or sulfated polysaccharides and salts which adsorb certain substances during direct blood perfusion treatment different from sulfated polysaccharides and salts thereof. A method for producing an adsorbent for use in direct blood perfusion, wherein a polysaccharide, a salt thereof, or a sulfated polysaccharide and a ligand different from the salt are bound to a water-insoluble carrier. (Iv) sulfated polysaccharides, salts thereof, or sulfated polysaccharides and salts thereof having an intrinsic viscosity of not less than 0.005 dl / g and not more than 0.5 dl / g of sulfur content of not less than 5% and not more than 22% by weight and (ii) direct blood perfusion A method for producing an adsorbent for use in direct blood perfusion, wherein a sulfated polysaccharide, a salt thereof, or a sulfated polysaccharide and a ligand different from the salt thereof is bound to a water-insoluble carrier during the treatment. 7. The sulfated polysaccharide, its salt, or sulfated polysaccharide and its salt are bound to the water-insoluble carrier according to claim 5 or 6 in an amount of 0.02 mg or more and 200 mg or less per ml of the water-insoluble carrier. Preparation of Adsorbents. 4. The adsorbent according to claim 3, wherein the water insoluble carrier has an average particle diameter of 30 mu m or more.