JPH06198167A - Bilirubin adsorbent - Google Patents

Abstract

PURPOSE:To obtain a safe adsorbent with which bilirubin in blood can be selectively adsorbed and removed by fixing a polymer containing three compds. expressed by specified general formulae and/or derivatives of these containing amino groups and imino groups to nonwoven fabric. CONSTITUTION:This bilirubin adsorbent is used for an indirect reflux method to selectively adsorb and remove bilirubin in blood for acute intoxication, fulminating hepatitis, etc. This adsorbent is obtd. by fixing the following polymer to nonwaven fabric. The polymer contains three kinds of compds. expressed by three general formulae and/or derived material of these containing amino groups and imino groups. In formulae, R<1>, R<4> are hydrogen atoms or methyl groups, R<2>, R<3>, R<5>, R<6> are hydrogen atoms or alkyl groups of 1-5 carbon number, R<7> is a hydrogen atom, alkyl group of 1-10 carbon number, or aryl group or may be branched by H-(NH-CH2CH2)q-. (n), (m) are integers 2-5, and (p), (q) are integers 2-200000.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a bilirubin adsorbent and an adsorbent. More specifically, the present invention relates to a bilirubin adsorbent and an adsorption device that selectively adsorb and remove bilirubin in blood.

[0002]

2. Description of the Related Art In cases such as acute drug poisoning and fulminant hepatitis,
Metabolic function declines, metabolic abnormal substances increase in blood, and in severe cases, jaundice, hepatic coma, etc. are induced. The cause of hepatic coma has not been fully clarified, but bilirubin is one of the indicators.
Bilirubin binds to cells and mitochondrial membranes and causes cell death. Furthermore, neuropathy, cerebral palsy, hearing loss,
It also causes a stroke. Conventionally, as a means for treating severe liver failure and hepatic coma, direct transfusion such as exchange transfusion and plasma exchange, and
An indirect perfusion method using activated carbon or the like is performed. The bilirubin adsorbent used in the indirect perfusion method includes activated carbon and styrene-divinylbenzene copolymer (AR-1, BR6).
01, Amberlite XAD-4), a tertiary amino anion exchange resin (IONEX), a strongly acidic anion exchange type ion exchange fiber (QAS) and the like.

The bilirubin adsorbents already applied for are porous copolymers of crosslinkable monomers and monovinyl monomers (JP-A-54-69289), hydroxyl groups, amino groups and / or Alternatively, a polymer having an imino group (JP-A-57-64059), a porous copolymer of a crosslinkable polymerizable monomer and a cyano-containing monomer (JP-A-58-29134).
No.), bilirubin adsorbent obtained by surface-treating porous particles composed of a copolymer of divinylbenzene and glycidyl methacrylate with plasma protein or a hydrophilic synthetic polymer (JP-A-63-267436), divinyl having a porous structure. An organic compound having an affinity for bilirubin was immobilized on the surface of a porous body composed of a copolymer of benzene and glycidyl methacrylate (JP-A-63-305935) and a copolymer of divinylbenzene and glycidyl methacrylate. Bilirubin adsorbent (JP-A-63-315145
No.), a copolymer having an anti-thrombogenicity, which comprises a copolymer of a (meth) acrylic acid ester oligomer having a hydroxyl group at the terminal (JP-A-2-132110), divinylbenzene, glycidyl methacrylate and hydroxyalkyl (meth). There is a bilirubin adsorbent (JP-A-2-286173) in which an organic compound having an affinity for bilirubin is immobilized on the surface of a porous terpolymer of acrylate.

However, none of these adsorbents has a good selective adsorptivity for bilirubin, and it is difficult to sufficiently deal with hyperbilirubinemia such as severe jaundice. Furthermore, there is a bilirubin adsorbent which is not necessarily safe from the viewpoint of blood compatibility and antithrombotic property. Therefore, a large amount of bilirubin,
It is desired to develop a safe bilirubin adsorbent that can adsorb efficiently and has high blood compatibility. The present inventors have conducted intensive studies in order to develop an adsorbent capable of specifically adsorbing bilirubin in blood. As a result, an amino group, a non-woven fabric having a polymer containing an imino group immobilized thereon, The inventors have found that bilirubin is efficiently adsorbed, and have reached the present invention.

[0005]

SUMMARY OF THE INVENTION Therefore, the object of the present invention is to provide a novel bilirubin adsorbent. The present invention also has a gist of the invention in that bilirubin can be selectively adsorbed and removed from human plasma, and bilirubin can be safely adsorbed and removed even in whole blood without separating blood cell components and plasma. Usually, when trying to remove bilirubin using a bead carrier, after filling the column with beads, blood is passed through the column to remove harmful substances in blood, but in this case, the pressure loss is large,
When whole blood is used, hemolysis of red blood cells occurs,
The current situation is that activation of complement and coagulation factors is caused, blood cell adhesion and blood coagulation occur, and safe and efficient adsorption and removal cannot be performed. An object of the present invention is to provide an adsorbent and an adsorbing device that have excellent blood compatibility and can adsorb and remove bilirubin efficiently even in whole blood treatment.

[0006]

[Means for Solving the Problems] The above-mentioned object is to immobilize a compound represented by Chemical formula 1, Chemical formula 2, Chemical formula 3 and / or a polymer containing amino groups and imino groups derived therefrom, on a non-woven fabric. Is achieved with a bilirubin adsorbent. The material of the nonwoven fabric used in the present invention,
Polyethylene terephthalate (PET), polypropylene (PP), polyethylene (PE), polyamide (N
y), cotton, rayon (Ry) and the like, but PET, PE, cotton and Ry are preferable in view of the strength retention after modification.

The fiber diameter used for these non-woven fabrics is
0.5 μm to 50 μm, preferably 1.0 μm to 3
In view of 0.0 μm, blood cell adhesion prevention and bilirubin adsorption capacity, 2.0 μm to 20.0 μm is particularly preferable. Weight per m ^{2 of} non-woven fabric used (unit weight)
Is 10.0 to 300.0 g, preferably 20.0 g
˜200.0 g, more preferably 30.0 g to 15
It is 0.0 g, and the thickness of one nonwoven fabric is 0.1 mm to 1
0.0 mm, preferably 0.2 mm to 7.0 mm, and in actual use, several non-woven fabrics are laminated and further pressed to adjust to a desired thickness and packed in an adsorption column. It is preferable.

There are various methods for introducing an amino group and an imino group into these nonwoven fabrics by using the compounds represented by Chemical formulas 1, 2, and 3. (A) In the case of PP or PE, the compounds represented by Chemical Formulas 1 and 2 are added during the polymerization of PP or PE, and the copolymerized polymer is spun to produce a non-woven fabric. Or P containing an imino group
A method of obtaining a nonwoven fabric of E. (B) The compounds represented by the general formulas 1 and 2 or solutions of these are applied to these nonwoven fabrics, and the electron beam (E
B), a method in which the compound represented by the general formula 1 or 2 is graft-polymerized by exposing it to ultraviolet rays (UV) or ozone to generate radicals or ions. When the compounds represented by the general formulas 1 and 2 are applied to these non-woven fabrics, it is preferable to apply using a solvent that dissolves these volatile compounds from the viewpoint of uniform application. The volatile solvent is not limited, but water, methanol, ethanol, methylene chloride, chloroform, acetone, dioxane, tetrahydrofuran, or a mixed solvent thereof is preferable. But,
Even when the compounds represented by the general formulas 1 and 2 (hereinafter abbreviated as vinyl compounds) are applied using such a method, if the boiling point of the vinyl compound is low, it will evaporate under dry conditions and the amount of grafting can be controlled. At the same time, it may become impossible to graft.

In order to suppress the volatilization of these vinyl compounds and improve the grafting efficiency, it is preferable to add a high-boiling point compound to the coating solution and coat the solution to suppress the evaporation of the vinyl compound. The high boiling point compound to be added is not particularly limited, but polyhydric alcohols such as ethylene glycol, diethylene glycol, polyethylene glycol having a degree of polymerization of 10 or less, and glycerin are particularly preferable. The addition amount of these high boiling point compounds is not particularly limited, but is 0.3% by weight ratio to the vinyl compound.
About 10 times the amount is preferable. As a method of further improving the grafting efficiency of the vinyl compound rather than the addition of the high boiling point compound, the addition of the crosslinkable vinyl compound is preferable. Examples of these crosslinkable vinyl compounds include methylenebisacrylamide, trimethylolpropane diacrylate, triallyl isocyanurate, trimethylolpropane diacrylate, tetramethylolmethane tetraacrylate, and other compounds having a plurality of vinyl groups in one molecule. Examples include compounds represented by general formula 4.

[Chemical 4] (In the formula, R ⁸ , R ⁹ , and R ¹⁰ each represent a hydrogen atom or a methyl group, and S represents an integer of 1 to 1000.) The addition amount of these crosslinkable vinyl compounds is a weight ratio to the vinyl compound. 1/50 to 50/1, preferably 1/30 to 3
It is 0/1. Among these crosslinkable vinyl compounds, the compound represented by the general formula 4 has a high solubility of the vinyl compound, can be uniformly applied, and has a high grafting efficiency. In addition, after grafting, a polyethylene glycol unit is introduced on the fiber surface. By making it hydrophilic, it suppresses adsorption of other useful components such as plasma proteins such as albumin and globulin when adsorbing and removing bilirubin in blood, and also suppresses blood cell adhesion such as leukocytes and platelets. However, it is particularly preferable because it has excellent blood compatibility.

The concentration of the solution for coating the nonwoven fabric with the mixture of the vinyl compound and the high boiling point compound or the crosslinkable vinyl compound is 0.1 to 20.0% by weight, preferably 0.1%.
It is 5 to 10.0% by weight. The solvent is not particularly limited as long as it dissolves both the vinyl compound and the crosslinkable vinyl compound or the high boiling point compound, but the above solvents are particularly preferable. As described above, as a method of coating the nonwoven fabric after coating the vinyl compound and the crosslinkable vinyl compound, there is a method of exposing to EB, UV or ozone as described above, but a method of grafting by EB irradiation is It is particularly preferable because it is simple and causes little decrease in strength of the material. The irradiation dose is preferably 1 to 50 Mrad,
It is 2 to 20 Mrad.

As described above, in addition to the method of applying a vinyl compound in the state of a monomer to a nonwoven fabric and then grafting it,
Homopolymers of the compounds represented by the general formulas 1 and 2 or copolymers with other vinyl monomers, and further polymers represented by the general formula 3 (hereinafter abbreviated as PEI)
It is also possible to graft onto a non-woven fabric by using. In order to graft a polymer or PEI derived from the compounds represented by the general formulas 1 and 2, it is essential to add the above-mentioned crosslinkable vinyl compound, and any polymer alone is subjected to EB irradiation. However, it cannot be grafted. Also in this case, it is essential to add the compound represented by the general formula 4 to the crosslinkable vinyl compound, and any of the polymers alone cannot be grafted by EB irradiation. Also in the crosslinkable vinyl compound in this case, the use of the compound represented by the general formula 4 suppresses non-specific adsorption and blood cell adhesion of useful blood components at the time of grafting and use after grafting, Particularly preferred. The other vinyl monomer referred to here corresponds to all vinyl monomers generally present, but by improving the hydrophilicity of the copolymer,
It is particularly preferable to use hydrophilic monomers such as acrylamide, hydroxyethyl methacrylate, N-isopropylacrylamide, acrylic acid, vinyl alcohol, etc., since useful plasma proteins and blood cell adhesion are suppressed. In these copolymers, the content of the compounds represented by the general formulas 1 and 2 is 2 mol% or more,
It is preferably 10 mol% or more.

Homopolymers derived from the compounds represented by the general formulas 1 and 2 or copolymers with other vinyl monomers and polymers represented by the general formulas 3 are used without using a crosslinkable vinyl compound. The grafting method, the amino group or imino group in the polymer, allyl chloride, a compound having both a vinyl group and a halide group, such as vinylbenzyl halide, alkylated amino group or imino group, After acidifying an amino group or an imino group in a polymer with an acid halide having a vinyl group such as acryloyl halide, dissolving it in the above volatile solvent, coating it on a non-woven fabric, drying and EB irradiation. Can be grafted. The concentration of the solution and the irradiation dose of EB are as described above.

As described above, there are various methods for introducing an amino group or an imino group into a non-woven fabric, but a homopolymer derived from the compounds represented by the general formulas 1 and 2 or another vinyl compound is used. The method of grafting onto the copolymerized polymer in the presence of the crosslinkable vinyl compound represented by the general formula 4 is an extremely excellent method because of high grafting efficiency and high blood compatibility of the resulting grafted nonwoven fabric. The content of the amino group or imino group thus introduced is 1 μeq / g to 5 meq / g, preferably 100 μeq / g to 4 meq / g, and more preferably 300 μeq / g to 3 meq / g. The non-woven fabric introduced with amino groups or imino groups obtained as above retains excellent bilirubin adsorption performance, non-specific adsorption of useful components in blood is also suppressed, and is useful as a bilirubin adsorbent. is there.

[0014]

EXAMPLES The present invention will be described in more detail with reference to the following examples. <Example 1> - Preparation of amino group-introduced nonwoven - polyethyleneimine _{(- (CH 2 -CH 2 -NH} )
_n −) 2 g and PEG-DAR (CH ₂ ═CH—COO
_{(CH 2 CH 2 O) 14} -OC-CH = CH 2) 0.6g
And are dissolved in 200 ml of methanol and the fiber diameter is 3.5 μm.
6 pieces (1.0 g / sheet) of non-woven fabric (PET-SPS: 15 cm × 12 cm) are soaked and air-dried, and 2 EB5Mrads are applied.
Irradiated twice, washed with distilled water 3 times and with methanol 3 times.
By the above operation, the amino group in the non-woven fabric is 0.611 me
q / g was introduced. --Adsorption test of bilirubin-- In a centrifuge tube containing 20 ml of bovine plasma in which bilirubin was dissolved, 0.5 g of the prepared non-woven fabric was cut into 1 mm squares and shaken in a 37 ° C. thermostat for 1 hour. The same operation was performed using only bovine plasma without using a non-woven fabric as a control. For the treated plasma, the total bilirubin and direct bilirubin were quantified using a bilirubin B2 test (manufactured by Wako Pure Chemical Industries, Ltd.) kit. The results are shown in Table 1. As shown in Table 1, this nonwoven fabric adsorbed bilirubin better than the nonwoven fabrics of Comparative Example 1 and Comparative Example 2.

[0015]

[Table 1]

[0016] <Example 2> - Preparation of amino group-introduced nonwoven - polyethyleneimine _{(- (CH 2 -CH 2 -NH} )
_n −) 2 g and PEG-DAR (CH ₂ ═CH—COO
(CH ₂ CH ₂ O) ₁₄ —OC—CH═CH ₂ ) 2 g and 200 ml of methanol are dissolved, and a nonwoven fabric (PET-SPS: 15 cm × 12 cm) having a fiber diameter of 3.5 μm 6 sheets (1.0 g / sheet) ), Air dry, and add EB5Mrad to 2
Irradiated twice, washed with distilled water 3 times and with methanol 3 times.
By the above operation, the amino group in the nonwoven fabric is 1.132me.
q / g was introduced. --Adsorption test of bilirubin-- In a centrifuge tube containing 20 ml of bovine plasma in which bilirubin was dissolved, 0.5 g of the prepared non-woven fabric was cut into 1 mm squares and shaken in a 37 ° C. thermostat for 1 hour. The same operation was performed using only bovine plasma without using a non-woven fabric as a control. For the treated plasma, the total bilirubin and direct bilirubin were quantified using a bilirubin B2 test (manufactured by Wako Pure Chemical Industries, Ltd.) kit. The results are shown in Table 1. As shown in Table 1, this nonwoven fabric adsorbed bilirubin better than the nonwoven fabrics of Comparative Example 1 and Comparative Example 2.

Comparative Example 1--Preparation of Polyethyleneimine-Introduced Nonwoven Fabric--Polyethyleneglycol-diacrylate (CH ₂ ═CH—CO (OCH ₂ CH ₂ ) ₃ —O—OC
CH = CH ₂ ) 3 g and methylenebisacrylamide ((CH ₂ ═CHCONH) ₂ CH ₂ ) 0.5 g were dissolved in 150 ml of methanol, and 6 sheets of non-woven fabric (PET-SPS: 15 cm × 12 cm) having a fiber diameter of 3.5 μm (1.
(0 g / sheet) was immersed, air-dried, irradiated with EB5Mrad twice, and washed with distilled water three times and methanol three times. --Adsorption test of bilirubin-- In a centrifuge tube containing 20 ml of bovine plasma in which bilirubin was dissolved, 0.5 g of the prepared non-woven fabric was cut into 1 mm squares and shaken in a 37 ° C. thermostat for 1 hour. The same operation was performed using only bovine plasma without using a non-woven fabric as a control. For the treated plasma, the total bilirubin and direct bilirubin were quantified using a bilirubin B2 test (manufactured by Wako Pure Chemical Industries, Ltd.) kit. The results are shown in Table 1. As shown in Table 1, this non-woven fabric hardly adsorbed bilirubin as compared with the non-woven fabrics of Example 1 and Example 2.

<Comparative Example 2> --Adsorption test of bilirubin-- A non-woven fabric (PET-SPS) having no functional group introduced into a centrifuge tube containing 20 ml of bovine plasma in which bilirubin was dissolved
0.5 g was cut into 1 mm square and put in it, and shaken in a constant temperature bath at 37 ° C. for 1 hour. The same operation was performed using only bovine plasma without using a non-woven fabric as a control. For the treated plasma, the total bilirubin and direct bilirubin were quantified using a bilirubin B2 test (manufactured by Wako Pure Chemical Industries, Ltd.) kit. The results are shown in Table 1. As shown in Table 1, this nonwoven fabric is
Compared with the nonwoven fabrics of Example 1 and Example 2, bilirubin was hardly adsorbed.

<Example 3> --Plasma protein adsorption test-- 0.5 g of the nonwoven fabric used in Examples 1 and 2 was placed in a centrifuge tube containing 20 ml of bovine plasma in which bilirubin was dissolved.
Was cut into 1 mm square, and shaken in a constant temperature bath at 37 ° C. for 1 hour. The same operation was performed using only bovine plasma without using a non-woven fabric as a control. Regarding the treated plasma, a protein quantification kit (BCA Protein Assay Re
Plasma proteins were quantified using an agent: BCA), an albumin quantification kit (Albumin B Test Wako: Wako Pure Chemical Industries, Ltd.), and C3 and C4 quantification plates (Hoechst). Plasma (non-woven fabric) was used as a control (before starting). The results are shown in Table 2. As shown in Table 2, the nonwoven fabrics used in Example 1 and Example 2 were
Almost no plasma proteins were adsorbed.

[0020]

[Table 2] In Tables 1 and 2, * 1 and * indicate units of mg / g.

As is clear from the above results, the bilirubin adsorbent of the present invention can selectively and easily remove bilirubin in blood and has little effect on other plasma components such as total protein, albumin and complement. I knew that.

[0022]

Since the bilirubin adsorbent of the present invention has good blood compatibility, it is not necessary to take a complicated method of separating a plasma component from blood and performing perfusion in advance,
A sufficient effect can be expected by directly perfusing blood. Therefore, it is widely used for removing bilirubin in blood by a simple operation.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Masakazu Tanaka 2-1-1 Katata, Otsu City, Shiga Prefecture Toyobo Co., Ltd.

Claims (1)

[Claims] 1. A compound represented by the following chemical formula 1, chemical formula 2, and chemical formula 3.
And / or amino groups and derivatives derived therefrom
A polymer containing mino groups is immobilized on a non-woven fabric.
Bilirubin adsorbent (in the formula, R¹, R^FourIs a hydrogen atom or
Methyl group, R ², R³, R^FiveAnd R⁶Is a hydrogen atom
Or an alkyl group having 1 to 5 carbon atoms or R²And R³, R
^FiveAnd R⁶May each form a heterocycle. R₇
Is a hydrogen atom, an alkyl group having 1 to 10 carbon atoms, an aryl group
Or- (NH-CH₂CH₂)_q-Even if branched
good. n and m each represent an integer of 2 to 5, and p and
And q are integers of 2 to 200,000, respectively.
You ). [Chemical 1][Chemical 2][Chemical 3]