WO2012124619A1

WO2012124619A1 - Polyanion-immobilized polymer substrate for removing viruses, and method for removing viruses

Info

Publication number: WO2012124619A1
Application number: PCT/JP2012/056068
Authority: WO
Inventors: 直人櫻井; 俊徳西山; 典孝吉川; 哲朗鈴木
Original assignee: Dic株式会社; 国立大学法人浜松医科大学
Priority date: 2011-03-11
Filing date: 2012-03-09
Publication date: 2012-09-20

Abstract

The present invention provides a polyanion-immobilized polymer substrate for removing viruses, said polyanion-immobilized polymer substrate enabling efficient removal of a virus from a liquid, and a method for removing viruses. In the case of applying to blood of a living organism, in particular, the present invention provides a method whereby a virus can be removed with low invasiveness, i.e., needing only a small amount of the blood to be taken out from the body and removing little useful blood components, and whereby the operation time can be shortened. The polyanion-immobilized polymer substrate for removing viruses according to the present invention is usable as a module that has a function of efficiently removing viruses and a function of not removing useful plasma components.

Description

Polymer base material for removing polyanion-immobilized virus, and method for removing virus

The present invention relates to a polymer substrate for removing a polyanion-immobilized virus and a method for removing a virus.

Hepatitis C is caused by chronic infection with hepatitis C virus (HCV), and a combination therapy of peginterferon and ribavirin is common as a therapeutic method using drugs. In patients with genotype 1b and a large amount of virus in the blood, the therapeutic result is about 50%, and the rate of transition to cirrhosis and liver cancer is high, so the development of more effective treatments and drugs is desired. (Non-Patent Document 1). In general, in the treatment with drugs, it is known that when the amount of virus in the blood is low, the treatment results are high, and when HCV in the blood is removed with a porous filter and combined therapy with the drug is performed, There is a report that treatment results are improved (Non-patent Document 2). That is, it is presumed that the treatment results have been improved by reducing the amount of virus in the body.

Further, in Patent Document 1, a blood inlet, an upstream blood circuit, a plasma separation unit, and a downstream blood circuit are connected in this order, and further, a plasma outlet of the plasma separation unit, an upstream plasma circuit, a plasma purification unit, and a downstream plasma The circuits are connected in this order, and the end of the downstream plasma circuit is a blood processing apparatus connected to blood plasma mixing means provided in the middle of the downstream blood circuit, and the blood plasma mixing means of the downstream blood circuit A device is described in which a blood cell treatment means comprising a water-insoluble carrier for removing at least viruses and virus-infected cells is provided on the downstream side, and the plasma purification means comprises a porous filtration membrane having a maximum pore diameter of 20 nm to 50 nm.

However, the method of removing with the above filter is to remove the virus from the plasma components after separating blood cells and plasma once, so the circuit configuration is complicated, and a method of removing the virus from the blood more easily is desired. Yes.

As an adsorbent for blood purification for hepatitis C virus to which a ligand or the like is immobilized, Patent Document 2 discloses that a peptide having affinity for immunoglobulin or the like is immobilized on a water-insoluble gel, and an immune complex type hepatitis C virus is An efficient removal method has been reported.

On the other hand, it is known that heparin is effective as a ligand that binds to HCV (Non-patent Document 3). Heparin is a kind of heparan sulfate that is chemically a glycosaminoglycan. It is a polymer in which β-D-glucuronic acid or α-L-iduronic acid and D-glucosamine are polymerized by 1,4-bonds. Thus, it is known as a polyanionic saccharide having a higher degree of sulfation than heparan sulfate.
Therefore, heparin is immobilized in a substrate in which heparin is immobilized on a polymer support such as a hollow fiber that can pass whole blood without separating blood cells and plasma, or in the pores of a blood cell plasma separation membrane. It is expected that HCV can be removed more easily with such a base material, and an HCV removal module or the like with less burden on the patient can be provided. Alternatively, the same effect can be expected if a base material on which polyanions having an action equivalent to that of heparin is immobilized.

Examples of the form of the substrate on which the polyanion is immobilized include beads and porous hollow fibers. The internal circulation type and filtration type extracorporeal circulation modules using porous hollow fibers have less blood retention than the extracorporeal circulation modules filled with the particulate polyanion-immobilized base material, Has the advantage of less formation. When the polyanion is immobilized on the porous hollow fiber, the type of surface functional group and the immobilization density vary depending on the material of the substrate, and it is necessary to find an optimum method for each substrate.

On the other hand, for a base material having an adsorption action for human immunodeficiency virus (hereinafter referred to as HIV), for example, Patent Document 3 discloses a polymer base material having a methylene group in the main chain, an ethylenically unsaturated bond and a sugar chain. Or a polymerizable composition containing the polymerizable compound is contacted and irradiated with ionizing radiation, or the polymeric base material is irradiated with ionizing radiation, and then the polymerizable compound or A sugar chain-immobilized polymer substrate for HIV adsorption having a sugar chain, obtained by contacting a polymerizable composition containing the polymerizable compound, is described.

JP-A-2005-230165 JP-A-10-323387 JP 2010-68910

In view of the prior art, the problem of the present invention is that a body fluid such as blood can flow without causing clogging due to generation of a thrombus and the like, and a substrate capable of efficiently removing viruses in the body fluid, To provide an instrument.

In order to solve the above problems, the present inventors have selected a polymer support, a method for immobilizing a polyanion having a function of removing a virus on a polymer support, and a polymer group on which the polyanion is immobilized. As a result of examining the materials in detail, the present invention has been completed.

That is, the present invention is a virus-removing polymer base material on which a polyanion is immobilized, in particular, the polymer base material is a polyolefin hollow fiber, and the polyanion is immobilized on the surface of the hollow fiber. It relates to a polymer substrate for virus removal having

ADVANTAGE OF THE INVENTION According to this invention, the polymeric base material which has the function which can selectively remove a virus, without adsorbing and removing the blood component which removal is not preferable, and a medical device using the same can be provided.

It is a schematic sectional drawing which shows an example of the medical device provided with the polymer base material of this invention.

That is, the present invention
1. Polyacrylic acid, polymethacrylic acid, polyitaconic acid, polymaleic acid, ring-opened polymer containing maleic anhydride polymer, polyfumaric acid, polyglutamic acid, polyaspartic acid, polyallylcarboxylic acid, polyvinylsulfuric acid, polyphosphoric acid, polyvinylphosphoric acid , Polyvinyl sulfonic acid, polystyrene sulfonic acid, polyallyl sulfonic acid, polyallyl phosphoric acid, 2- (meth) acryloyloxyethane sulfonate polymer, 2- (meth) acryloyloxy ethane phosphate polymer, 2-acrylamide-2 A polymer substrate for virus removal in which a polyanion selected from methylpropanesulfonic acid polymer and polyisoprenesulfonic acid polymer is immobilized;
2.1. In the polymer substrate for virus removal to which the polyanion described in is immobilized,
A polymer group for virus removal, wherein the polyanion is immobilized on a polymer support (A) surface-treated with a polymer material having a hydroxyl group via a compound (B) having a group capable of reacting with a hydroxyl group and a polyanion. Material,
3. Polymer support (A) surface-treated with a polymer material having a hydroxyl group is a partially saponified product of ethylene-vinyl alcohol copolymer, ethylene-vinyl alcohol-vinyl acetate copolymer, ethylene-vinyl acetate copolymer 1. A hollow fiber surface-treated with a vinyl alcohol-vinyl acetate copolymer, a hydroxymethacrylate copolymer, a partially saponified cellulose acetate, or a glycerin derivative. A polymer substrate for virus removal according to claim 1,
4). The compound (B) having a group capable of reacting with a hydroxyl group and a polyanion is epichlorohydrin or a diepoxy compound. Or 3. A polymer substrate for virus removal according to claim 1,
5.1. In the polymer substrate for virus removal to which the polyanion described in is immobilized,
A polymer substrate for virus removal in which the polyanion is immobilized on a polymer support (C) by graft polymerization;
6.1. In the polymer substrate for virus removal to which the polyanion described in is immobilized,
A polymer substrate for virus removal, wherein the polyanion is immobilized by surface-treating the polymer support (C) with the polyanion,
7. 4. The polymer support (C) is a hollow fiber. Or 6. A polymer substrate for virus removal according to claim 1,
8). 2. The hollow fiber is a porous hollow fiber. Or 7. A polymer substrate for virus removal according to claim 1,
9. 7. The hollow fiber is based on polyethylene, polypropylene or poly-4-methylpentene A polymer substrate for virus removal according to claim 1,
10. 7. The average flow pore size of the porous hollow fiber is in the range of 10 to 500 nm. Or 9. A polymer substrate for virus removal according to claim 1,
11. 7. The inner diameter of the porous hollow fiber is in the range of 150 to 500 μm. ~ 10. A polymer substrate for virus removal according to any one of
12 7. The film thickness of the porous hollow fiber is in the range of 30 to 100 μm. ~ 11. A polymer substrate for virus removal according to any one of
13. 7. The amount of polyanion immobilized on the porous hollow fiber is in the range of 1 to 100 μg / cm ² . ~ 12. A polymer substrate for virus removal according to any one of
14 1. The virus is hepatitis B or C virus ~ 13. A polymer substrate for virus removal according to any one of
15. Claim 1. ~ 14. A virus removal method using the polymer substrate for virus removal according to any one of
16.15. In the virus removal method described in the above, it has a step of mixing the liquid that has passed through the holes of the hollow fiber and the liquid that has not passed through the holes by passing the virus-containing liquid through the porous hollow fiber. Virus removal method,
17. The fluid containing virus is blood containing virus. The virus removal method as described in 1. above.

Hereinafter, the present invention will be described in detail.
Polyanion The polyanion used in the present invention is not particularly limited as long as it is a polymer repeatedly containing an anionic functional group such as a carboxyl group, a sulfonic acid group, a sulfuric acid group, and a phosphoric acid group. Examples of such polymers include polyacrylic acid, polymethacrylic acid, polyitaconic acid, polymaleic acid, maleic anhydride polymer-containing polymer ring-opened product, polyfumaric acid, polyglutamic acid, polyaspartic acid, polyallylcarboxylic acid, polyvinyl sulfate, Polyphosphoric acid, polyvinyl phosphoric acid, polyvinyl sulfonic acid, polystyrene sulfonic acid, polyallyl sulfonic acid, polyallyl phosphoric acid, 2- (meth) acryloyloxyethane sulfonate polymer, 2- (meth) acryloyloxyethane phosphate polymer And 2-acrylamido-2-methylpropanesulfonic acid polymer, polyisoprenesulfonic acid polymer, and the like. In the case of surface graft polymerization, the corresponding monomer may be reacted.

-Porous hollow fiber The porous hollow fiber used for this invention will not be restrict | limited especially if prepared by a well-known and usual method. Examples include olefin resins such as polyethylene, polypropylene, and poly-4-methylpentene, styrene resins, sulfone resins, acrylic resins, urethane resins, ester resins, ether resins, and cellulose mixed esters. More specifically, polyethylene terephthalate, polymethyl methacrylate, polysulfone, polyether sulfone, polyacrylonitrile and the like can be mentioned. What is necessary is just to manufacture a porous hollow fiber by a well-known and usual method according to the intended purpose. In the case of a polyolefin hollow fiber, those having various pore diameters and pore diameter distributions can be prepared by subjecting the spun yarn to annealing treatment, cold drawing, hot drawing, and heat setting.

-Graft polymerization to porous hollow fiber The graft polymerization reaction used in the present invention may be a known method such as a method using ionizing radiation irradiation or a method using a radical generating reagent. The method using ionizing radiation irradiation can be said to be an environmentally friendly polymerization reaction because no solvent or reagent is used when radicals are generated.

-Graft polymerization method by ionizing radiation irradiation In the present invention, in particular, a method of graft polymerization by reacting radicals generated by irradiating the hollow fiber with ionizing radiation and an ethylenically unsaturated group of a vinyl monomer or the like. Can be illustrated. Examples of the ionizing radiation source used in the graft polymerization include known and commonly used α-rays, β-rays, γ-rays, accelerated electron beams, X-rays and the like, and practically preferred are γ-rays and accelerated electron beams. When actually performed, the simultaneous irradiation graft polymerization method in which the porous hollow fiber and the vinyl monomer are brought into contact with each other and irradiated with ionizing radiation, and after the hollow fiber has been previously irradiated with ionizing radiation and before being brought into contact with the vinyl monomer. Any of the irradiation graft polymerization methods is possible and can be selected according to the purpose.

In the graft polymerization method using ionizing radiation used in the present invention, since the irradiation dose and the acceleration voltage differ depending on the hollow fiber, it is not possible to determine the range in general, and appropriate considering the material, form, thickness, etc. of the hollow fiber It is necessary to adjust. For example, when the irradiation amount is large, dielectric breakdown due to charging occurs, and when the irradiation amount is small, the polymerization reaction does not proceed. For this reason, in consideration of the material and form of the hollow fiber, the amount of irradiation that does not cause dielectric breakdown due to charging and the polymerization reaction proceeds sufficiently may be adjusted as appropriate. Further, the acceleration voltage is related to the permeability and varies depending on the thickness of the hollow fiber. In the case of a thin form such as a film, the acceleration voltage is generally small and may be selected depending on the form of the polymer support.

For example, in the case of a hollow fiber having a thickness of 10 (μm) to 100 (μm) made of poly-4-methylpentene, the hollow fiber is 10 (kGy) or more and 300 (kGy) or less, and more preferably 90 (KGy) or less, and the acceleration voltage can be selected as appropriate.

In the irradiation graft polymerization, radicals in the substrate after irradiation are quickly inactivated by temperature increase and contact with oxygen. Therefore, after irradiation, it is preferable to store at a low temperature in a state where oxygen is sufficiently removed, and to carry out the polymerization reaction promptly. For the above reasons, the polymerization reaction is preferably carried out under deoxygenation or under an inert gas.

The hollow fiber after the graft polymerization reaction may be removed from unreacted monomers and homopolymers by various methods such as washing with a solvent.

In addition, when polymerizing the monomer to the hollow fiber, a polymerization reaction can be performed on either the inner surface or the outer surface of the yarn, or both, depending on the embodiment. For example, when the blood is perfused inside the hollow fiber, the monomer solution may be contacted only inside the hollow fiber. When the blood is perfused outside the hollow fiber, the monomer solution is contacted only outside the hollow fiber. It ’s fine. In order to pass through the hole of the hollow fiber, the monomer solution may be brought into contact with the inside of the hole for polymerization.

-Graft polymerization reaction by radical generating reagent In this invention, the polymerization method of the porous hollow fiber surface-treated with the polymeric material which has a hydroxyl group can be illustrated especially. In this case, it is possible to use diammonium cerium nitrate (CAN), which is known and commonly used as a radical generating method for polyvinyl alcohol and cellulose, as an initiator. The surface-treated porous hollow fiber is immersed in a vinyl monomer solution, and CAN is put in a reaction vessel in the form of a solution or a solid and reacted for a predetermined time. The concentration of the vinyl monomer or CAN may be appropriately adjusted according to the target graft ratio, and the monomer concentration may generally be in the range of 1 to 200 mg / mL, and CAN may be in the range of 0.1 to 100 mM. The reaction temperature may be about room temperature to 100 ° C.
In order to immobilize the polyanion by generating a radical and performing a graft polymerization reaction, as described above, the polymerization reaction is performed by generating a radical at the hydroxyl group of the polymer support (A) surface-treated with the polymer material having a hydroxyl group. Alternatively, polymerization may be performed by generating radicals directly on the surface of a hollow fiber made of polyolefin such as polyethylene, polypropylene or poly-4-methylpentene.

-Polymeric material having a hydroxyl group Examples of the polymeric material having a hydroxyl group that can be used in the present invention include an ethylene-vinyl alcohol copolymer, an ethylene-vinyl alcohol-vinyl acetate copolymer, and an ethylene-vinyl acetate copolymer. Examples include partially saponified products of polymers, those containing vinyl alcohol copolymers such as vinyl alcohol-vinyl acetate copolymers, those containing hydroxy methacrylate copolymers, partially saponified products of cellulose acetate, or glycerin derivatives. it can. However, in addition to those exemplified, there is no particular limitation as long as the resin has a hydroxyl group.
The surface treatment with the polymer material having a hydroxyl group can be performed by a known and usual method. For example, after the porous polyolefin is immersed in a solution in which the polymer material having a hydroxyl group is dissolved and pulled up, Drying and the like can be mentioned as a preferable method. Among these, an ethylene-vinyl alcohol copolymer is preferable in that the polyolefin porous hollow fiber can be easily hydrophilized as disclosed in JP-A-61-271003. Alternatively, it is also possible to use a method of making a porous mixture of polyolefin or the like and a polymer material having a hydroxyl group in advance.

Compound having a group capable of reacting with hydroxyl group and polyanion (B)
In the present invention, a compound capable of reacting with a polyanion is immobilized on a hollow fiber surface-treated with a polymer material having a hydroxyl group, and the polyanion can be immobilized via the compound. In particular, a diepoxy compound bonded with epichlorohydrin, an alkylene group or a phenylene group is preferable (hereinafter referred to as an epoxy compound). Here, as the diepoxy compound, ethylene glycol diglycidyl ether, 1,4-butanediol diglycidyl ether, neopentyl glycol diglycidyl ether, bisphenol A diglycidyl ether, 1,3-butanediene diepoxide, 1,5 -Hexadiene diepoxide and the like. These compounds can be immobilized on the porous hollow fiber by a known and conventional method. Furthermore, since an epoxy group remains, a polyanion can be immobilized through the epoxy group, which is preferable. In addition, when fixing an epoxy compound to this hollow fiber, according to embodiment, it can fix to either the inner surface of a thread | yarn, an outer surface, or both. For example, when blood is perfused inside the hollow fiber, an epoxy compound may be brought into contact with the inside of the hollow fiber, and conversely, during external perfusion, an epoxy compound may be brought into contact with the outside of the hollow fiber. When trying to pass through the hole of the hollow fiber, the epoxy compound may be brought into contact with the inside of the hole and fixed.

The reaction between the remaining epoxy group and the polyanion may be performed by a known and conventional method. A solution in which a polyanion is dissolved may be prepared and reacted as it is, or a reaction may be performed by adding a catalytic amount of an acid, a base, a Lewis acid or the like. The reaction solution, temperature, time, concentration, catalyst, and the like may be appropriately selected depending on the type of polyanion and the substrate, and are not particularly limited as long as the polyanion can be immobilized. *

-Surface treatment of polymer support (C) with polyanion In the present invention, a method of immobilizing a polyanion on the surface by treating a porous raw hollow fiber with a polyanion-containing polymer material is also included. As a treatment method, a preferable method is to immerse the porous hollow fiber in a solution in which the polymer material is dissolved, pull it up and dry it. For example, when a polyanion-containing resin is immobilized on a polyolefin hollow fiber, it may be a polyanion formed solely from an anionic monomer, or may be a copolymer in consideration of the solubility of the resin and the adhesion to the polyolefin hollow fiber. Also good. As such a resin, if it is a polymer formed from a single monomer, polyacrylic acid, polymethacrylic acid, polyitaconic acid, polymaleic acid, a ring-opened polymer of a maleic anhydride polymer, polyfumaric acid, polyglutamic acid, poly Aspartic acid, polyallylcarboxylic acid, polyvinylsulfuric acid, polyphosphoric acid, polyvinylphosphoric acid, polyvinylsulfonic acid, polystyrene sulfonic acid, polyallylsulfonic acid, polyallylphosphoric acid, 2- (meth) acryloyloxyethane sulfonate polymer, 2- Examples include (meth) acryloyloxyethane phosphate polymer, 2-acrylamido-2-methylpropanesulfonic acid polymer, and polyisoprenesulfonic acid polymer.

On the other hand, for copolymers, ethylene-anionic monomer copolymer, butadiene-anionic monomer copolymer, methyl acrylate-anionic monomer copolymer, ethyl acrylate-anionic monomer copolymer, butyl acrylate- An anionic monomer copolymer etc. are mentioned, A meta (acrylic acid), a styrenesulfonic acid, etc. can be illustrated as a preferable anionic monomer especially. In addition, when processing this hollow fiber with a polyanion containing polymeric material, it can fix to either the inner surface of a thread | yarn, an outer surface, or both according to embodiment. For example, when blood is perfused into the hollow fiber, a polymer material may be fixed inside the hollow fiber, and conversely, during external perfusion, the polymer material may be brought into contact with the outside of the hollow fiber. When trying to pass through the hole of the hollow fiber, the polymer material may be brought into contact with the inside of the hole and fixed.
In the present invention, a post-processed post-processed porous hollow fiber is illustrated, but it is also possible to use a method of making a porous mixture of polyolefin or the like and a polyanion or polyanion-containing polymer in advance. .

As a method of removing the virus, it is possible to remove the virus from the liquid by immersing and contacting the polyanion-immobilized hollow fiber in a liquid containing the virus, but when using a porous hollow fiber, By passing the virus-containing liquid through the hole of the hollow fiber, the virus can be efficiently removed. Considering the case of processing blood during extracorporeal circulation, it is convenient and desirable that whole blood can be processed in the pores, but high biocompatibility is required because of the problem of stagnation and blood cells directly contacting the pores.

On the other hand, a method of separating blood cells and plasma components, allowing only the plasma components to pass through the pores, and removing viruses from the plasma can be mentioned. In that case, a liquid that has passed through the holes of the hollow fiber and a liquid that has not passed through the holes are produced. From the examination of the virus removal rate in the liquid containing virus, the virus removal rate in the liquid that passed through the hole of the hollow fiber is a good result as described in the following examples, and is a useful component in the blood It became clear that albumin was not removed. Here, passing through the hole means a state in which liquid passes through from the hollow inner surface to the outer surface side or from the outer surface to the inner surface side.

The pore diameter of the porous hollow fiber used is not particularly limited as long as it has a diameter capable of efficiently removing the virus. For example, in order to efficiently remove viruses from plasma by extracorporeal circulation, it is necessary to design as follows. When blood cells and plasma are separated and virus is removed from the plasma, it must have a function as a plasma separation membrane. Therefore, it is desirable that the mean flow pore size is 500 nm or less so that blood cell components and platelets do not enter the pores.

Furthermore, since it is necessary to design so that the permeability | transmittance of the protein component in plasma does not fall, 50 nm or more is desirable as an average flow hole diameter. Therefore, in order to have a function as a plasma separation membrane, it is necessary to design the average flow pore size to 50 to 500 nm. Since the present invention has a feature of efficiently removing viruses from plasma separated from blood, the optimal pore size varies depending on the size of the virus when virus removal is assumed.
Taking hepatitis C virus as an example, the preferred pore size is 80 to 250 nm, more preferably 100 to 180 nm. In the case of a relatively large human immunodeficiency virus, the preferable pore size is 100 to 250 nm, more preferably 120 to 200 nm. When the polyanion-immobilized hollow fiber is to be removed by immersing it in a virus-containing liquid, there is no particular limitation, and the pore diameter may be adjusted according to the purpose.

The inner diameter of the porous hollow fiber used is not particularly limited as long as it has an inner diameter capable of efficiently removing the virus. For example, when using a hollow fiber in extracorporeal circulation, it is necessary to design as follows.

Since the amount of blood that can be circulated from humans is limited, the size of the circulation module and the like cannot be excessively increased. When the inner diameter is large, the number of yarns that can be put into the module is reduced, so that the contact area may be reduced or the linear velocity may be inferior and blood may be retained.

On the other hand, if the inner diameter becomes too small, the blood cell component is likely to be clogged. Considering these points, the inner diameter is preferably 150 to 500 μm, more preferably 160 to 400 μm, and further preferably 170 to 350 μm.

The thickness of the porous hollow fiber used is not particularly limited as long as it has a thickness capable of efficiently removing the virus. For example, when the virus is efficiently removed from the plasma by extracorporeal circulation, it is preferably 30 to 100 μm, more preferably 35 to 80 μm, taking into consideration the plasma separation performance, contact area, mechanical strength of the hollow fiber, etc. More preferably, the thickness is 40 to 60 μm.

Furthermore, it is also possible to improve the virus removal rate by combining another substrate having a function of capturing and removing viruses outside the hollow fiber. Such other substrate is not particularly limited as long as it has a function of capturing and removing viruses, and examples thereof include a material in which a compound capable of adsorbing viruses is immobilized on a gel or a nonwoven fabric. it can.
The amount of polyanion immobilized on the porous hollow fiber used in the present invention is preferably in the range of 1 to 100 μg / cm ² in view of efficiently removing viruses.

-Liquid containing virus The liquid containing virus targeted in the present invention is not particularly limited as long as it contains a virus. More specifically, for example, a body fluid which is a human body fluid component, a culture solution containing a virus, and the like can be mentioned. More specific examples of body fluids include blood, saliva, sweat, urine, runny nose, semen, plasma, lymph, tissue fluid and the like.

The form of the medical device comprising the polymer base material of the present invention is not particularly limited as long as it is a shape applicable to the above-mentioned use, and examples thereof include a hollow fiber module, a filtration column, and a filter. . In the hollow fiber module and the filtration column, the shape and material of the container are not particularly limited, but when applied to extracorporeal circulation of body fluid (blood), a cylindrical container having an internal volume of 10 to 400 mL and an outer diameter of about 2 to 10 cm It is preferable to use a cylindrical container having an internal volume of 20 to 300 mL and an outer diameter of about 2.5 to 7 cm. An example is shown in FIG.

As a method for using the medical device of the present invention, any method may be used as long as it can be removed from and separated from the virus containing the virus.

The following examples illustrate the invention in more detail.

<Pore diameter of porous polymer substrate>
In accordance with ASTM F316-86 and ASTM E1294-89, Porous Materials, Inc. Using a “Palm Porometer CFP-200AEX” manufactured by the company, the average flow pore size (average pore size of the constricted portion of the hole penetrating from one side of the membrane to the other) was measured by the half dry method. Perfluoropolyester (trade name “Galwick”) was used as the test solution.

The pores do not necessarily have to penetrate the membrane as a straight tube, and may be bent inside the membrane. Also, some holes may be fused inside the membrane, or conversely, one hole may be branched, or these may be mixed.

<Polyanion immobilized amount>
The amount of polyanion immobilized on the hollow fiber was determined by weight increase or titration.
<HCV removal test>
A hollow fiber module with a membrane area of 1.8 cm ² was prepared, 0.6 mL of plasma (stock solution) of HCV patient was passed through, 0.3 mL of liquid (filtrate) that passed through the hole, and liquid that did not pass through the hole ( (Inner liquid) 0.3 mL was obtained. Specimen was measured by ortho HCV antigen ELISA test,
HCV removal rate (%) = (1-HCV amount in filtrate / HCV amount in stock solution) × 100
Asked.

<ELISA>
The specimen is pretreated with a pretreatment solution (SDS) to release the HCV core antigen and simultaneously deactivate the coexisting HCV antibody to obtain a measurement sample. A measurement sample is added to an HCV core antigen antibody-immobilized plate and incubated. After the reaction for a predetermined time, washing is performed, and whole radish-derived peroxidase-labeled HCV core antigen antibody is added and incubated. After reacting for a predetermined time, washing is performed, and o-phenylenediamine reagent is added and incubated. After the reaction for a predetermined time, a reaction stop solution is added. Measure the color development at a wavelength of 492 nm. The concentration is calculated from the absorbance of the sample.

<Plasma albumin permeation amount>
Bromocresol green reagent is added to the specimen, and color development is measured at a wavelength of 630 nm. The concentration was calculated from the absorbance of the sample.
Albumin permeability (%) = (albumin content in filtrate / albumin content before filtration) × 100

-Preparation example of porous hollow fiber High density polyethylene (HIZEX 2200J manufactured by Mitsui Petrochemical Co., Ltd.) having a density of 0.968 g / cm ³ and a melt index of 5.5 is used. Using a spinneret for hollow fiber shaping having a discharge cross-sectional area of 1.06 cm ² , spinning was performed at a spinning temperature of 160 ° C. and wound up with a spinning draft 1427. The dimensions of the obtained unstretched hollow fiber were an inner diameter of 308 μm and a film thickness of 64 μm. This unstretched hollow fiber was heat-treated at 115 ° C. for 24 hours at a constant length. Subsequently, the film was stretched 1.8 times at a deformation rate of 21400% / min at room temperature, and then heated in a heating furnace at 100 ° C. until the total stretching ratio was 4.8 times so that the deformation rate was 330% / min. Further, the film was further subjected to thermal shrinkage in a heating furnace at 125 ° C. until the total draw ratio became 2.8 times to obtain a porous raw drawn yarn.

Example of surface treatment with a polymer material having a hydroxyl group When treating with a polymer material having a hydroxyl group, for example, an ethylene vinyl alcohol copolymer (EVOH) having an ethylene content of 44% is heated and dissolved in a 75% aqueous ethanol solution. As a result, a solution having a concentration of 2.5% by weight was obtained. The above-mentioned porous raw stretched yarn was immersed in the solution kept at 50 ° C. for 100 seconds, kept at 80 ° C. under ethanol saturated steam at 50 ° C. for 80 seconds, and then the solvent was further dried over 80 seconds. The obtained EVOH hydrophilized porous hollow fiber membrane had an inner diameter of 287 μm and a film thickness of 52 μm.

Example 1 Immobilization of Polyanion to Polyethylene Porous Hollow Fiber According to JP-A-2-0259260, 100 mL of acetone, 80 mL of epichlorohydrin, 21 mL of 40% NaOH aqueous solution were put in a test tube, an inner diameter of 257 μm, a film thickness of 54 μm, an average An EVOH hydrophilized porous hollow fiber membrane having a pore size of 183 nm was immersed. The reaction was carried out at 30 to 40 ° C. for 5 hours while applying ultrasonic waves. After completion of the reaction, the reaction product was washed with acetone and water to obtain a hollow fiber having an epoxy group introduced therein. Next, a 10% aqueous solution of polyacrylic acid having a molecular weight of 5000 was prepared, and the epoxy-introduced hollow fiber was immersed and reacted at 40 ° C. for 3 days. From the value of weight increase, 31 μg / cm ² (in terms of inner surface area) was immobilized as polyacrylic acid. When the plasma of an HCV patient was filtered with the module obtained using the above-mentioned polyacrylic acid-immobilized hollow fiber membrane, 53% of HCV was removed. At this time, the albumin permeability was 98%.

(Comparative Example 1)
When the same evaluation was performed using the EVOH hydrophilized hollow fiber membrane in which the polyacrylic acid used in Example 1 was not immobilized, the HCV removal rate was 19%. Therefore, it was found that the hollow fiber in which polyacrylic acid having a molecular weight of 5000 is immobilized can improve the removal rate of HCV without removing biological components such as albumin.

(Example 2)
Polyacrylic acid having a molecular weight of 25000 was immobilized in the same manner as in Example 1 except that the molecular weight of the polyacrylic acid in Example 1 was changed to 25000. From the value of weight increase, 73 μg / cm ² (in terms of inner surface area) was fixed as polyacrylic acid. When the plasma of an HCV patient was filtered with a module obtained by using the above-described polyacrylic acid-immobilized hollow fiber membrane, 63% of HCV was removed. At this time, the albumin permeability was 98%.

(Comparative Example 2)
When the same evaluation was performed using the EVOH hydrophilized hollow fiber membrane in which the polyacrylic acid used in Example 1 was not immobilized, the HCV removal rate was 19%. Therefore, it was found that the hollow fiber in which polyacrylic acid having a molecular weight of 25000 is immobilized can improve the removal rate of HCV without removing biological components such as albumin.

(Example 3) Production of acrylic acid grafted hollow fiber 1.5 g of acrylic acid monomer was dissolved in 100 mL of ethyl acetate, and dissolved oxygen in the solution was removed by reducing the pressure while stirring on an ice bath.
On the other hand, a porous raw hollow fiber bundle made of poly-4-methylpentene (hollow fiber inner surface area: 146 cm ² , manufactured by DIC Corporation) is irradiated with an electron beam of 90 kGy, placed in a glass test tube and sealed with a rubber stopper. The inside of the test tube was evacuated.

Next, a deoxygenated acrylic acid monomer solution was added to a hollow fiber bundle-filled test tube irradiated with an electron beam to start graft polymerization. After standing for 1 hour, the hollow fiber bundle was taken out, and washing with methanol and water was repeated until the unreacted monomer and the like were below the detection limit (1 μg / mL, the same applies hereinafter) by GPC measurement. Thus, the properties of the resulting acrylic acid polymer-bonded hollow fiber were an acrylic acid polymer bond amount of 26 mg and a bond density (in monomer conversion) of 2.5 μmol / cm ² (inner surface area conversion).

Example 4
In the same manner as in Example 3, methacrylic acid was graft-polymerized on the porous raw hollow fiber to obtain a methacrylic acid grafted hollow fiber having a bond density (in terms of monomer) of 2.5 μmol / cm ² (in terms of internal surface area).

(Comparative Example 3)
Hydroxyethyl methacrylate (HEMA) was graft polymerized in the same manner as in Example 3 to obtain a HEMA graft hollow fiber having a bond density (in terms of monomer) of 2.7 μmol / cm ² (in terms of internal surface area).
<HCV removal test by immersion>
The grafted poly-4-methylpentene hollow fiber was cut into 5 cm ² , put in a microtube, added with 500 μL of HCV patient serum diluted with PBS, and rotated at 23 ° C. for 60 minutes. The amount of HCV core antigen before and after rotation was measured by ELISA, and the HCV removal rate was measured. The removal rate was 56% for the acrylic acid grafted hollow fiber (Example 3) and 48% for the methacrylic acid grafted hollow fiber (Example 4).
On the other hand, in the hollow fiber grafted with HEMA of Comparative Example 3, the removal rate was 9%.

(Example 5)
A 200 mL three-necked flask was charged with 60 mL of a 0.5N nitric acid aqueous solution and 2.5 g of acrylic acid, and an EVOH hydrophilized porous membrane (inner diameter 257 μm, film thickness 54 μm) was immersed therein. After the atmosphere in the flask was replaced with nitrogen on an ice bath, 100 mg of diammonium cerium nitrate was dissolved and reacted at 80 ° C. for 1 hour. After the reaction, unreacted acrylic acid and homopolymer were washed with water. The target polyacrylic acid-immobilized hollow fiber was obtained with a graft ratio of 2.7%, calculated from the weight increase of the hollow fiber. When the plasma of an HCV patient was filtered with a module obtained by using the acrylic acid grafted hollow fiber membrane, 66% of HCV was removed. At this time, the albumin permeability was 99% or more.

(Comparative Example 4)
When the same evaluation was performed using the EVOH hydrophilized hollow fiber membrane not grafted with acrylic acid used in Example 5, the HCV removal rate was 17%. Therefore, it was found that the hollow fiber grafted with acrylic acid can improve the HCV removal rate without removing biological components such as albumin.

(Example 6)
Acrylic acid was grafted in the same manner as in Example 5 except that the amount of acrylic acid in Example 5 was changed to 0.8 g. The graft ratio at this time was 0.4%. When the plasma of an HCV patient was filtered with a module obtained using the acrylic acid grafted hollow fiber membrane, 56% of HCV was removed. At this time, the albumin permeability was 97%.

(Example 7)
An ethylene-acrylic acid copolymer (acrylic acid 15 wt%) was dissolved in heated toluene to prepare a 2.5 wt% solution. The solution was kept warm to such an extent that the resin did not precipitate, the porous stretched yarn was immersed, pulled up and dried to obtain a hollow fiber having acrylic acid fixed on the surface. When the plasma of HCV patients was filtered with a module obtained using the above-described surface acrylic acid-immobilized hollow fiber membrane, 30% of HCV was removed. At this time, the albumin permeability was 98% or more.

The instrument using the hollow fiber of the present invention can be used for removing viruses such as hepatitis virus.

1: Inlet containing virus 2: Outlet of virus not passing through hole 3: Hollow fiber membrane 4: Outlet of virus passing through hole 5: Container 6: Septum

Claims

Polyacrylic acid, polymethacrylic acid, polyitaconic acid, polymaleic acid, ring-opened polymer containing maleic anhydride polymer, polyfumaric acid, polyglutamic acid, polyaspartic acid, polyallylcarboxylic acid, polyvinylsulfuric acid, polyphosphoric acid, polyvinylphosphoric acid , Polyvinyl sulfonic acid, polystyrene sulfonic acid, polyallyl sulfonic acid, polyallyl phosphoric acid, 2- (meth) acryloyloxyethane sulfonate polymer, 2- (meth) acryloyloxy ethane phosphate polymer, 2-acrylamide-2 A polymer substrate for virus removal, on which a polyanion selected from a methylpropanesulfonic acid polymer and a polyisoprenesulfonic acid polymer is immobilized;
In the polymer substrate for virus removal to which the polyanion according to claim 1 is immobilized,
A polymer group for virus removal, wherein the polyanion is immobilized on a polymer support (A) surface-treated with a polymer material having a hydroxyl group via a compound (B) having a group capable of reacting with a hydroxyl group and a polyanion. Wood.
Polymer support (A) surface-treated with a polymer material having a hydroxyl group is a partially saponified product of ethylene-vinyl alcohol copolymer, ethylene-vinyl alcohol-vinyl acetate copolymer, ethylene-vinyl acetate copolymer The polymer substrate for virus removal according to claim 2, which is a hollow fiber surface-treated with a vinyl alcohol-vinyl acetate copolymer, a hydroxymethacrylate copolymer, a partially saponified product of cellulose acetate, or a glycerin derivative.
The polymer substrate for virus removal according to claim 2 or 3, wherein the compound (B) having a group capable of reacting with a hydroxyl group and a polyanion is epichlorohydrin or a diepoxy compound.
In the polymer substrate for virus removal to which the polyanion according to claim 1 is immobilized,
A polymer substrate for virus removal, wherein the polyanion is immobilized on a polymer support (C) by a graft polymerization reaction.
In the polymer substrate for virus removal to which the polyanion according to claim 1 is immobilized,
A polymer substrate for virus removal, wherein the polyanion is immobilized by surface-treating the polymer support (C) with the polyanion.
The polymer substrate for virus removal according to claim 5 or 6, wherein the polymer support (C) is a hollow fiber.
The polymer substrate for virus removal according to claim 3 or 7, wherein the hollow fiber is a porous hollow fiber.
9. The polymer substrate for virus removal according to claim 8, wherein the hollow fiber is made of polyethylene, polypropylene or poly-4-methylpentene as a substrate.
The polymer substrate for virus removal according to claim 8 or 9, wherein an average flow pore size of the porous hollow fiber is in the range of 10 to 500 nm.
The virus-removable polymer substrate according to any one of claims 8 to 10, wherein the inner diameter of the porous hollow fiber is in the range of 150 to 500 µm.
The polymer substrate for virus removal according to any one of claims 8 to 11, wherein the porous hollow fiber has a thickness of 30 to 100 µm.
The polymer substrate for virus removal according to any one of claims 8 to 12, wherein the amount of polyanion immobilized on the porous hollow fiber is in the range of 1 to 100 µg / cm 2 .
The polymer substrate for virus removal according to any one of claims 1 to 13, wherein the virus is a hepatitis B or C virus.
A virus removal method using the virus-removable polymer substrate according to any one of claims 1 to 14.
The virus removal method according to claim 15, comprising a step of mixing the liquid that has passed through the holes of the hollow fiber and the liquid that has not passed through the holes by passing the virus-containing liquid through the porous hollow fiber. A virus removal method characterized by the above.
The method for removing a virus according to claim 16, wherein the liquid containing the virus is blood containing the virus.