JPH05245198A - Leucocyte capturing body and its production - Google Patents

Abstract

PURPOSE:To provide the leucocyte capturing body required for removing leucocytes from blood components and the process for production of this capturing body. CONSTITUTION:This leucocyte capturing body consists of the leucocyte capturing body formed by introducing at least one kind of functional groups by a plasma initiated graft polymn. method to at least a part of the surface of a base material constituting the leucocyte capturing body and bonding these functional groups and a cationic material having the section reactable therewith. As a result, the secure introduction of the cationic polymer having a leucocyte capturing ability on the surface of the base material is possible and further, the capturing of the leucocytes while obtaining the stable filtration flow rate is possible without intrusion of foreign matters and the supply of means for medical treatment having no problems at all in terms of safety is possible.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a leukocyte capturing body required for removing leukocytes from blood components and a method for producing the same. More specifically, it is possible to capture leukocytes by imparting a cationic property to the surface of the capturing body by the plasma-initiated graft polymerization method without mixing foreign substances into the blood components after leukocyte removal and obtaining a stable filtration flow rate. The present invention relates to a body and a method for manufacturing the body.

[0002]

2. Description of the Related Art In principle, currently performed component transfusion is to transfuse only blood components necessary for a patient and not unnecessary components. However, while the concentrated red blood cells that are used on a daily basis are said to be component preparations, in reality many white blood cells,
Blood diseases with a high frequency of blood transfusion of these concentrated red blood cells, including platelets and plasma (malemic anemia, hemolytic anemia, malignant lymphoma, etc.), malignant tumor, renal failure, infection,
In collagen disease, chronic hemorrhagic anemia, etc., in order to suppress the production of antibodies against white blood cells and platelets, and for patients who develop side effects of blood transfusion, the white blood cells and platelets contained in concentrated red blood cells are removed to prevent it and the purity is high. Need to transfuse red blood cells. Recently, attention has been focused on Graft-versus-Host Disease (GVHD), in which mitogenic lymphocytes in the transfused blood recognize and recognize the patient's histocompatibility antibody as a foreign substance, and the use of leukocyte-depleted blood products has been noticed. It is increasing.

Therefore, it is necessary to rupture the mitotic ability of lymphocytes in blood for transfusion, and the widely known method at this time is to rupture the ability of lymphocytes to divide themselves by radiation. Examples include capturing leukocytes (lymphocytes) by filtration.

In the method using radiation, blood products containing lymphocytes (fresh blood, preserved blood, red blood cell concentrate, concentrated platelets, etc.) are irradiated with radiation of 1500 rad (15 Gy),
It destroys the ability of lymphocytes to divide. In Japan,
The irradiation of the blood bag is performed in the radiation treatment room mainly by an X-ray technician, and one batch is completed in about 5 to 6 minutes.
In Europe and the United States, a dedicated irradiation device for blood products that uses Cs ¹³⁷ as a radiation source is commercially available. 400-500 ml with this
The blood bag can be irradiated in 2 to 6 minutes. According to Leitman et al., Transfusion 25 (4): 293-300,1985, this dose did not impair the functions of platelets and granulocytes as well as red blood cells.
However, expensive equipment and a management system by the person in charge of handling radiation are indispensable for equipment installation, and not everyone can operate it. It is also possible that transfusion is not possible if the patient is in emergency use at night and the risk is high.

On the other hand, with regard to the capture of leukocytes by filtration, the operation is very easy, it is inexpensive and anyone can operate it. This takes advantage of the characteristic of being physically adsorbed on a substance having a small curvature unique to white blood cells, and is obtained by compressing and laminating a porous body or laminating fibers having a small diameter. However, since it physically captures leukocytes, it has a problem that reliability such as radiation treatment is lacking.

According to Zierdt et al. (Applied and E
nviromental Microbiology, 1979, 12, 1166-1172), bacteria, erythrocytes, leukocytes, polystyrene spheres, etc. Have found things.
Zierdt et al. Treated the membrane with a nonionic surfactant and confirmed that the particles were not trapped, concluding that the trapping was due to electrostatic interactions.
In addition, Zierdt et al., Solvent cast membranes like nylon have a strong surface charge,
On the other hand, it has been confirmed that ordinary fibrous cellulosic materials which are not solvent-cast have a small surface charge.

From the above, it is suggested that the surface should have a strong surface charge (Zeta-potential is positive) in order to capture leukocytes, and since it is possible to chemically capture, leukocyte Even if it is expected that the clogging of the pores is expected, the pore diameter can be expected to increase.

As a specific surface modification method, for example, in US Pat. Nos. 3,242,073 and 3,352,424, a conventional anion type filter medium having an adsorbed cationic organic polyelectrolyte is used to remove microorganisms from a liquid. It is described to be removed. It is possible to trap particles with a negative surface charge on this filter media. U.S. Pat. No. 4,178,438 describes a method for purifying industrial effluents using a cationically modified cellulosic material. Also, U.S. Pat. No. 3,808,305.
The publication describes the charge obtained by a liquid having a matrix polymer, a polyelectrolyte and a crosslinker. The solvent is evaporated from the cast film and the film is later chemically crosslinked to insolubilize it. Further, U.S. Pat. Nos. 3,944,485 and 4,045,352 describe an ion exchange resin obtained by preliminarily allowing a monomer to be present on a fiber to produce an ion exchange hollow fiber, and then polymerizing the monomer.

However, only adsorbing the cationic polyelectrolyte on the anion type filter medium has a problem in safety such as desorption, peeling and elution, and it is not preferable as a medical device and some insolubilization operation is required. I have to do it. Further, when the base material is a poorly soluble material such as polyolefin, polyester, or polyamide that composes the filter medium, the solvent is limited, and high temperature and high pressure are required. In many cases.

[0010]

When a substance which is hardly soluble or hardly compatible composes the base material as described above, there are restrictions on the solvent, and there are risks such as high temperature and high pressure. It was a thing. Further, the coating has a problem in safety, and the operation after the coating becomes complicated in the crosslinking.

Therefore, the object of the present invention is to solve the above-mentioned problems, there is almost no limitation on the type of base material, and a substance having leukocyte-capturing ability is chemically bound to the surface of the base material. Another object of the present invention is to provide a leukocyte-capturing body which can be expected to be suitable for use as a medical instrument and a method for producing the same.

[0012]

Means for Solving the Problems As a result of intensive studies on the leukocyte-capturing body and a method for producing the same, the present inventors have introduced a functional group onto the surface of a substrate by a plasma-initiated graft polymerization method, It has been found that the above objects can be achieved by chemically binding to a substance having a leukocyte-capturing ability, and the present invention has been completed based on this finding.

That is, the above-mentioned objects are as follows: (1) At least one kind of functional group is introduced into the surface of at least a part of the substrate constituting the leukocyte-capturing body by the plasma-initiated graft polymerization method, and these functional groups are combined. It is achieved by a leukocyte-capturing body characterized by being bound with a cationic substance having a site capable of reacting.

Another object of the present invention is also achieved by the leukocyte-capturing body according to (1) above, wherein (2) the cationic substance is a compound having a quaternary nitrogen in its main chain or side chain.

Another object of the present invention is (3) above, wherein the functional group is at least one selected from the group consisting of an acid halide group, an isocyanate group and an epoxy group. This is achieved by the leukocyte trapper described in 2).

Still another object of the present invention is (4) wherein the shape of the leukocyte-capturing body is a porous body, a flat membrane, a hollow fiber, a solid thread, a non-woven fabric, a woven fabric, a knitted fabric or a composite thereof. This is achieved by the leukocyte-capturing body according to any one of (1) to (3) above.

In the present invention, (5) the base material constituting the leukocyte-capturing body is polyolefin, halogenated polyolefin, polyester, polyurethane, polysulfone, polyether sulfone, poly (meth) acrylate, butadiene-acrylonitrile copolymer, polyamide, This is achieved by the leukocyte-capturing body according to any one of (1) to (4) above, which is an etheramide-polyamide block copolymer, an ethylene-vinyl alcohol copolymer, or a mixture thereof.

Another object of the present invention is to (6) introduce at least one functional group into the surface of at least a part of the base material constituting the leukocyte-capturing body by a plasma-initiated graft polymerization method, and to introduce these functional groups. It can also be achieved by a method for producing a leukocyte trap, which comprises binding a cationic substance having a site capable of reacting with

Another object of the present invention is also achieved by the method for producing a leukocyte-capturing body according to the above (6), wherein (7) the cationic substance is a compound having a quaternary nitrogen in its main chain or side chain. To be done.

Another object of the present invention is (8) above, wherein the functional group is at least one selected from the group consisting of an acid halide group, an isocyanate group and an epoxy group. This is achieved by the method for producing a leukocyte-capturing body according to 7).

Still another object of the present invention is that the cationic substance (9) has the general formula (1):

[0022]

[Chemical 1]

(In the formula, at least one of R ₁ , R ₂ and R ₃ has at least one functional group selected from the group consisting of an amino group, a carboxyl group and a hydroxyl group, and the rest of the moieties are alkyl groups or An alkyl group containing an ether bond, an ester bond, an amide bond, and likewise R ₅ , R ₆ , and R
_At least one of ₇ has at least one kind of functional group selected from the group consisting of an amino group, a carboxyl group and a hydroxyl group, and the remaining sites are an alkyl group or an ether bond,
An alkyl group containing an ester bond or an amide bond, R
₄ is an alkylene group or a benzyl group, an ether bond,
An alkylene group containing an ester bond and an amide bond,
X ⁻ represents a counter ion), which is achieved by the method for producing a leukocyte-capturing body according to any one of the above (6) to (8).

Another object of the present invention is (10) the shape of the leukocyte-capturing body is a porous body, a flat membrane, a hollow fiber, a solid thread, a non-woven fabric, a woven fabric, a knitted fabric or a composite thereof. This is achieved by the method for producing a leukocyte-capturing body according to any of (6) to (9) above.

Still another object of the present invention is that (11) the base material constituting the leukocyte-capturing body is polyolefin, halogenated polyolefin, polyester, polyurethane, polysulfone, polyethersulfone, poly (meth) acrylate, butadiene-acrylonitrile. This is achieved by the method for producing a leukocyte-capturing body according to any one of the above (6) to (10), which is a copolymer, a polyamide, an etheramide-polyamide block copolymer, an ethylene-vinyl alcohol copolymer, or a mixture thereof.

[0026]

The leukocyte-capturing substance used in the present invention has at least one functional group introduced into the surface of at least a part of the substrate constituting the leukocyte-capturing substance by a plasma-initiated graft polymerization method, and these functional groups are introduced. It is a leukocyte-capturing body obtained by binding a cationic substance having a site capable of reacting with.
As this functional group, —COOH, —NH ₃ , and —OH are also conceivable, but an acid halide group, an isocyanate group and an epoxy group, which have high reactivity, are preferable. The substance having these functional groups to be introduced is preferably any substance as long as it has at least one selected from the group consisting of an acid halide group, an isocyanate group and an epoxy group. Well, more specifically, (meth) acrylic acid halides such as (meth) acrylic acid chloride and (meth) acrylic acid bromide, 2- (meth) acryloyloxyethyl isocyanate, and 3- (meth) acryloyloxypropyl isocyanate. Nato, 4- (meth) acryloyloxybutyl isocyanate, (meth) acryloyloxyisopropyl isocyanate, 2- (meth) acryloyloxy-1-methylethyl isocyanate, 2- (meth) acryloyloxy-1-ethylethyl isocyanate Nato, 4- (meth) acryloyloxy-2 (Meth) acryloyloxyalkyl isocyanate such as ethyl butyl isocyanate, 2- (meth) acryloyloxy-2,2-dimethylethyl isocyanate, 2,3-epoxypropyl (meth) acrylate, 3,4-epoxybutyl ( (Meth) acrylate,
2,3-epoxyisobutyl (meth) acrylate,
Examples thereof include epoxy group-containing monomers such as 3,4-epoxy-1-methylbutyl (meth) acrylate and 2,3-epoxy-1-methylethyl acrylate. Further, two or more kinds of substances having these functional groups to be introduced may be introduced, and after introduction to the surface of the base material, chemical treatment is performed to convert them into acid halide groups, isocyanate groups, epoxy groups, etc. It may be combined with a cationic substance.
The cationic substance having a leukocyte-capturing ability may be any as long as it reacts with the functional group, but preferably,
Those having an acid halide group, an isocyanate group, an amino group that reacts with an epoxy group, a carboxyl group, and a hydroxyl group are desirable, and specifically, a compound having a quaternary nitrogen in the main chain or side chain is the main component. I wish I had it. For example, the general formula (1),

[0027]

[Chemical 1]

(In the formula, at least one of R ₁ , R ₂ and R ₃ has at least one functional group selected from the group consisting of an amino group, a carboxyl group and a hydroxyl group, and the remaining sites are alkyl groups or An alkyl group containing an ether bond, an ester bond, an amide bond, and similarly,
At least one of R ₅ , R ₆ and R ₇ is an amino group,
It has at least one kind of functional group selected from the group consisting of a carboxyl group and a hydroxyl group, and the remaining part is an alkyl group or an alkyl group containing an ether bond, an ester bond or an amide bond, and R ₄ is an alkylene group or a benzyl group. Group, an alkylene group containing an ether bond, an ester bond, an amide bond, and X ⁻ represents a counter ion).

Specific examples of the general formula (1) will be given below. N, N'-tetramethyl-di (2-hydroxy-3)
-Chloropropyl) -alkylenediamine dichloride,
N, N'-tetramethyl-di (2-carboxy-3-chloropropyl) -alkylenediamine dichloride, N,
N'-tetrapropyl-di (2,3-epoxy-2-chloromethyl)-(alkylenediamine dichloride, N,
N'-Tetramethyl-di (2-hydroxy-3-chloropropyl) -alkylenediamine dibromide, N, N '
-Tetrachloromethyl-di (2-carboxy-3-chloropropyl) -alkylenediamine dichloride, N,
N'-tetramethyl-di (3-hydroxy-2-chlorobutyl) -alkylenediamine chloride, N, N'-tetramethyl-di (2,3-epoxy-2-chloro-3-
Bromopropyl) -alkylenediamine dichloride and the like. Other cationic substances include (meth) acrylic acid- (meth) acryloyloxyethyldiethylmethylammonium methylsulfate copolymers,
Copolymer of (meth) acrylic acid-quaternary ammonium ester of (meth) acrylic acid and (meth) acrylic acid such as a copolymer of (meth) acryloyloxypropyldiethylmethylammonium methylsulfate and guanidine in the main chain, Examples thereof include polymers containing ethyleneimine and imidazole and salts thereof.

Further, these cationic polymers having leukocyte-capturing ability are contained in an amount of 0.1 to 30% by weight based on the base material.
Preferably 0.5 to 20% by weight, more preferably 1 to 1
It is preferably 5% by weight.

Furthermore, the present invention introduces at least one of an acid halide group, an isocyanate group and an epoxy group into the surface of at least a part of the base material constituting the leukocyte-capturing body by a plasma-initiated graft polymerization method, and these The present invention relates to a method for producing a leukocyte-capturing body, which comprises binding a cationic substance having a site capable of reacting with a functional group. In order to introduce an acid halide group, an isocyanate group or an epoxy group, plasma is irradiated in the presence of a monomer having a reactive group in the molecule to perform plasma polymerization on the base material or the surface of the base material. After radicals are generated in the polymer, plasma polymerization may be performed by using the radicals as polymerization initiation points. For example, in order to generate radicals on the surface of the base material, hydrogen abstraction agents, neutron irradiation, γ-ray irradiation, electron beam irradiation, plasma irradiation, etc. may be mentioned. Production is preferred. The gas used for plasma irradiation is not particularly limited and may be in the presence of a monomer gas, but plasma irradiation with an inert gas such as argon is preferable, and the pressure during irradiation is 0.01 to 10 torr, preferably 0.0
It is desirable to be 5 to 5 torr, more preferably 0.1 to 1 torr. The irradiation time may be any time as long as the physical properties of the substrate are not impaired, but preferably 1
Seconds to 2 minutes, more preferably 5 seconds to 1 minute, still more preferably 10 to 30 seconds.

Further, in order to bring the base material having a radical into contact with the substance having an acid halide group, an isocyanate group or an epoxy group, it may be directly contacted in the presence or absence of a solvent. In consideration of sex and safety,
It is preferable to supply the above-mentioned substance having a functional group in a gaseous state and to contact with a base material having a radical, and the pressure at the time of supply is 0.001 to 100 torr, preferably 0.01 to 5
It is desirable that the pressure is 0 torr, more preferably 0.1 to 10 torr. Also, the supply time is 1 from the viewpoint of operability.
It is desirable that the time is 0 second to 2 hours, preferably 30 seconds to 1 hour, and more preferably 1 minute to 30 minutes. In addition, it is not necessary to directly bond a substance having an acid halide group, an isocyanate group, or an epoxy group to the radicals generated on the surface of the base material. After introducing a substance having no group, a substance having an acid halide group, an isocyanate group or an epoxy group may be introduced to the surface of the substrate. Also,
In order to bind the cationic substance to the functional group-introduced substrate, known methods such as polymerization and dipping can be mentioned, and it is preferable to immerse the cationic substance in an appropriate solvent for a certain period of time.

Furthermore, the present invention is characterized in that the shape of the leukocyte trap is a porous body, a flat membrane, a hollow fiber, a solid thread, a non-woven fabric, a woven fabric, a knitted fabric, or a complex thereof. It is about. Specifically, a porous body, a flat membrane, a non-woven fabric, a woven fabric, a knitted fabric, or a composite thereof which can easily generate radicals by plasma irradiation is preferable. Further, the pore size suitable for capturing leukocytes while obtaining a stable filtration flow rate with this capturing body is 0.1 to 30 μm, preferably 1 to 25 μm, as measured by mercury porosimetry.
More preferably, the thickness is 2 to 20 μm.

Further, in the present invention, the base material constituting the leukocyte-capturing body is polyolefin, halogenated polyolefin, polyester, polyurethane, polysulfone, polyether sulfone, poly (meth) acrylate, butadiene-.
The present invention relates to a leukocyte trap, which is an acrylonitrile copolymer, a polyamide, an etheramide-polyamide block copolymer, an ethylene-vinyl alcohol copolymer, or a mixture thereof.
From the viewpoint of operation, it is preferably polyolefin, halogenated polyolefin, polyurethane, ethylene-vinyl alcohol copolymer or a mixture thereof.

[0035]

EXAMPLES As the material for the base material used in the present invention, one or more kinds of materials may be used as long as they are as described above. Further, the shape of the base material may be a single shape or a composite shape thereof as long as it is as described above.

Examples 1 to 4 Rubicell (polyurethane porous body manufactured by Toyo Polymer Co., Ltd.) was subjected to Soxhlet washing with methanol for 8 hours, sufficiently dried, and then low temperature plasma (argon, 0.2 torr).
Was irradiated for 20 seconds to carry out surface graft polymerization of various monomers shown in Table 1 at a temperature of 288K. Structural formula (2),

[0037]

[Chemical 2]

After dipping in a 5% by weight aqueous solution (containing 0.5% by weight of pyridine) of cathionone UK (a surface cationizing agent manufactured by Meisha Co., Ltd.) for 12 hours at 60 ° C. and thoroughly washing with ion-exchanged water, A sample was obtained after drying. When this sample was evaluated by IR and ESCA, the presence of a cationizing agent was confirmed on the surface, and this sample was confirmed to be a polyurethane porous body having a cationized surface. Also,
The average pore diameter was 8 μm.

The obtained leukocyte-capturing body was punched out to a diameter of 25 mm and filled in a dedicated module. On the other hand, C of a healthy person
CRC prepared from PD-added fresh blood (WBC: 3.0 x 1
0 ^{3 to} 5.0 × 10 ³ , PLT: 1.0 × 10 ^{5 to} 3.
0 × 10 ⁵ , RBC: 3.0 × 10 ^{6 to} 5.0 × 1
0 ⁶ ) was flown and the ability to capture leukocytes was observed. as a result,
It was confirmed that the cationized surface of the polyurethane porous body had an improved trapping ability as compared with the polyurethane porous body before the treatment. The results are shown in Table 1.

Examples 5 to 6 Rubicell (polyurethane porous material manufactured by Toyo Polymer Co., Ltd.) was subjected to Soxhlet washing with methanol for 8 hours, sufficiently dried, and then low temperature plasma (argon, 0.2 torr).
Was irradiated for 20 seconds to carry out surface graft polymerization of glycidyl acrylate at a temperature of 288K. 3 for the porous body
Seed cationizing agents (PANFIX PX, Eretat U
-52, all in one company's surface cationizing agent) in a 10% by weight aqueous solution (containing 0.5% by weight of pyridine) at 60 ° C., 1
The sample was obtained by immersing it for 2 hours, thoroughly washing it with ion-exchanged water, and then drying it.

When this sample was evaluated by IR and ESCA, the presence of a cationizing agent was confirmed on the surface, and it was confirmed that this sample was a polyurethane porous body whose surface was cationized. The average pore diameter was 8 μm.

The obtained leukocyte-capturing body was punched out to a diameter of 25 mm and filled in a dedicated module. On the other hand, C of a healthy person
CRC prepared from PD-added fresh blood (WBC: 3.0 x 1
0 ^{3 to} 5.0 × 10 ³ , PLT: 1.0 × 10 ^{5 to} 3.
0 × 10 ⁵ , RBC: 3.0 × 10 ^{6 to} 5.0 × 1
0 ⁶ ) was flown and the ability to capture leukocytes was observed. as a result,
It was confirmed that the cationized surface of the polyurethane porous body had an improved trapping ability as compared with the polyurethane porous body before the treatment. The results are shown in Table 2.

Comparative Example 1 Rubicell (polyurethane porous body manufactured by Toyo Polymer Co., Ltd.) was subjected to Soxhlet washing with methanol for 8 hours to remove existing impurities. The average pore size was 6 μm. This porous body was punched out to a diameter of 25 mm, filled in a dedicated module, and fresh human whole blood was flown to observe the leukocyte capturing ability, but it was not as captureable as the polyurethane porous body having a cationized surface. .. The results are shown in Table 1.
Shown in.

Comparative Example 2 The same porous material as in Example 1 was immersed in an aqueous cathionone UK solution under the same conditions without plasma treatment. The obtained porous body was punched out to a diameter of 25 mm, filled into a dedicated module, and fresh human whole blood was flowed to observe the leukocyte capturing ability. The polyurethane porous body having a cationized surface has a better capturing ability. There wasn't.

[0045]

[Table 1]

[0046]

[Table 2]

[0047]

EFFECT OF THE INVENTION By using the method for producing a leukocyte-capturing body according to the present invention, it becomes possible to firmly introduce a cationic polymer having leukocyte-capturing ability onto the surface of a substrate. Therefore, the leukocyte-capturing body obtained by the present invention is capable of capturing leukocytes while obtaining a stable filtration flow rate without mixing foreign matter, and can achieve an excellent leukocyte removal rate. It is possible to supply medical devices with no problems in terms of safety.

Claims (2)

[Claims] 1. A cationic substance having a site capable of reacting with at least one kind of functional group introduced into the surface of at least a part of a base material constituting a leukocyte trap by a plasma-initiated graft polymerization method. A leukocyte-capturing body which is bound to a substance. 2. A cationic material having a site capable of reacting with at least one kind of functional group introduced into the surface of at least a part of a base material constituting a leukocyte trap by a plasma-initiated graft polymerization method. A method for producing a leukocyte trap, which comprises binding a substance.