JP2002035118A - Column for treating inflammatory disease - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a column for treating inflammatory disease which can efficiently treat inflammatory diseases by selectively adsorbing a bacteria-derived toxin, such as the endotoxin, etc., and an inflammatory leukocyte directly from blood with high efficiency. SOLUTION: This column for treating inflammatory disease is filled up with an inflammatory agency adsorbent which can remove both the bacteria-derived toxin and inflammatory leukocyte. The adsorbent is composed of a water- insoluble polymer prepared by bonding N-substituted poly(alkylene imine) residues to side chains.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a column for treating both inflammatory diseases by removing both bacterial toxins and inflammatory leukocytes in blood.

[0002]

2. Description of the Related Art Ischemic reperfusion injury of organs, sepsis, ulcerative colitis, Crohn's disease, systemic inflammatory response syndrome (SIR)
S) It has long been known that inflammatory leukocytes such as granulocytes increase in blood in inflammatory diseases such as infectious diseases. Bacterial toxins such as endotoxin, peptidoglycan, lipoteichoic acid, superantigen, and enterotoxin act to activate leukocytes in blood and increase inflammatory leukocytes.

[0003] As a column for removing endotoxin, a column in which an adsorbent in which polymyxin is immobilized on polystyrene fibers is packed (Japanese Patent Application Laid-Open No. 60-209525).
However, they do not adsorb inflammatory leukocytes with high efficiency. Columns for removing inflammatory leukocytes include those packed with cellulose acetate beads (JP-A-2-193969) and those packed with a polyester nonwoven fabric (JP-A-60-193468). However, it cannot adsorb bacterial toxins such as endotoxin.

[0004] As described above, there is no known compound capable of simultaneously removing bacterial toxins and inflammatory leukocytes with high efficiency and being used for treatment of extracorporeal circulation. Further, even if these are used in combination, it is not preferable because the proportion of blood circulating outside the body increases, which increases the burden on the patient.

Therefore, the present inventors have paid attention to the fact that simultaneous removal is important for treating inflammatory diseases efficiently.

[0006] As described above, there is no known inflammatory protein and inflammatory leukocyte which can be simultaneously removed and can be used for extracorporeal circulation treatment.

[0007]

SUMMARY OF THE INVENTION In view of the background of the prior art, an object of the present invention is to selectively adsorb bacterial-derived toxins such as endotoxin and inflammatory leukocytes from blood directly and efficiently with high efficiency. An object of the present invention is to provide a column for treating inflammatory diseases, which can efficiently treat inflammatory diseases.

[0008]

The present invention employs the following means in order to solve the above-mentioned problems. That is, the column for treating inflammatory diseases of the present invention is a column filled with an inflammatory substance adsorbent capable of removing both toxins derived from bacteria in blood and inflammatory leukocytes.
As the inflammatory substance adsorbent, a water-insoluble polymer having an N-substituted poly (alkyleneimine) residue bonded to a side chain is used.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION The object of the present invention is to treat inflammatory diseases efficiently by selectively adsorbing toxins derived from bacteria such as endotoxin and inflammatory leukocytes with high efficiency from blood directly. A column for the treatment of inflammatory diseases that can be used has been studied diligently, and as such an inflammatory substance adsorbent, using a water-insoluble polymer to which a specific nitrogen-substituted residue is bound, It has been determined that both toxins and inflammatory leukocytes can be removed.

The bacterial toxin referred to in the present invention means endotoxin, peptidoglycan, lipoteichoic acid, superantigen, enterotoxin and the like.

The inflammatory leukocytes according to the present invention include granulocytes,
Monocytes, or activated lymphocytes.

An inflammatory substance adsorbent capable of simultaneously removing bacterial toxins and inflammatory leukocytes is provided by bonding an N-substituted poly (alkylimine) residue to a water-insoluble polymer by means such as grafting. can do.

The water-insoluble polymer is not particularly limited as long as it is insoluble in water and can immobilize an N-substituted poly (alkyleneimine) by a covalent bond. Examples thereof include aromatic polyvinyl compounds and polysulfone-based polymers. , Polyetherimide, polyimide, polyamide, polyether, polyphenylene sulfide and the like, and a water-insoluble polymer having a reactive functional group capable of immobilizing an N-alkylated polyalkyleneimine by a covalent bond.

As such a water-insoluble polymer, in particular,
Lisulfone-based polymers can be preferably used,
For example,-｛(p-C₆H_Four) -SO_Two− (P−C₆H
_Four) -O-poly (p-phenylene ethers)
Rufon) unit or-｛(p-C₆H_Four) -SO_Two− (P
-C₆H_Four) -O- (p-C₆H_Four) -C (CH_Three) _Two
− (P−C₆H_Four) -O｝-,-｛(p-C₆H_Four)-
SO_Two− (P−C₆H _Four) -O- (p-C₆H_Four)-
O｝-,-｛(p-C₆H_Four) -SO_Two− (P−C₆H
_Four) -O- (p-C₆H_Four) -C (CF_Three)_Two− (P−
C₆H_Four) -U｝ -polyether sulfone
Use a polymer represented by a unit, etc.
Can be.

The reactive functional group includes an active halogen group such as a halomethyl group, a haloacetyl group, a haloacetamidomethyl group, a halogenated alkyl group, an epoxide group, a carboxyl group, an isocyanate group, a thioisocyanate group,
Acid anhydride groups and the like can be used,
Active halogen groups are easy to produce, highly reactive,
It is preferably used because the immobilization reaction of the N-substituted poly (alkylimine) residue can be performed under mild conditions and the covalent bond formed at this time is chemically stable.

Specific examples of the water-insoluble polymer having such a reactive functional group include chloroacetamidomethyl polystyrene, chloroacetamidomethylated Udel polysulfone, and chloroacetamidomethylated polyetherimide. . Further, among these water-insoluble polymers, those soluble in an organic solvent are particularly preferably used because they are easy to mold.

In the present invention, the N-substituted poly (alkyleneimine) means a poly (alkyleneimine) obtained by substituting the hydrogen of a nitrogen atom with at least one selected from an alkyl group and an acyl group. .

Examples of such poly (alkyleneimine) include polyethyleneimine, polypropyleneimine, polybutyleneimine, polyhexamethyleneimine,
A poly (ethylene imine / decamethylene imine) copolymer or the like can be used. Further, as the alkyl group as the N-substituent, n-butyl group, n-hexyl group,
An n-decanyl group, a lauryl group, a myristyl group, an n-hexadecyl group, an n-stearyl group, a benzyl group, and the like can be used.As the acyl group as the N-substituent, a butyroyl group, an n-hexinoyl group, An n-decanoyl group, a lauroyl group, a myristoyl group, an n-hexadecyloyl group, an n-stearyloyl group, a benzoyl group, and halogen-substituted products thereof can be used.

The adsorption performance and processability of the N-substituted poly (alkyleneimine) are determined by the length of the alkylene group in the main chain, the number of alkyl carbon atoms in the N-substituent, for example, the N-alkyl group, and It varies depending on the N-alkylation rate. The parameters include the ratio of the number of carbon atoms to the number of nitrogen atoms in the molecule as a whole (hereinafter abbreviated as C / N ratio),
It can be determined using the N-substitution rate. That is,
When the C / N ratio is too small, the hydrophilicity becomes too high,
If it is difficult to process, on the other hand, if it is too large, the adsorbing ability will decrease, so it is preferably 2.3 to 26, more preferably 2.
The range is preferably 5 to 14. The N-substitution rate is
Since the adsorption capacity is lowered even if it is too small or too large, it is preferably 1% or more and less than 30%, more preferably 1% or more and 20% or less. It is good from the function of removing both inflammatory leukocytes. In addition, the higher the degree of polymerization (n) of poly (alkyleneimine), the higher the adsorption performance to a high molecular weight substance is. However, if the degree of polymerization is too large, it is difficult to fix the poly (alkyleneimine) in the form of extending the graft chain. Therefore, preferably 4 to 100,000, more preferably 20 to 3
00 is good. Further, a part of the alkylene group in the main chain portion of the N-alkylated polyalkyleneimine may be substituted with a cyclohexane ring, a benzene ring, or the like, or may be substituted with an amide group, an ether group, or the like.

The immobilization density of the N-substituted poly (alkyleneimine) group in the adsorbent of the present invention depends on the C / N ratio of the N-substituted poly (alkyleneimine), the size of n, and the water-insoluble polymer serving as the trunk. Although it depends on the chemical structure and use of the polymer, if the amount is too small, the function does not appear.On the other hand, if the amount is too large, the moldability of the polymer after immobilization becomes poor, and the function as an adsorbent also decreases. The density is preferably 0.0001 per repeating unit of the water-insoluble polymer.
0.5 to 0.5 mol, more preferably 0.001 to 0.1 mol.

The method for preparing the N-substituted poly (alkyleneimine) is as follows.
It can be easily adjusted by mixing the required amount of the acid chloride or acid anhydride of the alkyl bromide or carboxylic acid at a temperature of 00 ° C. or less. This reaction proceeds quantitatively in a polar solvent such as ethyl alcohol, tetrahydrofuran or dimethylformamide even at room temperature. When the molecular weight of the poly (alkyleneimine) is high and the N-alkylation is high, the N-alkylated polyalkyleneimine becomes difficult to dissolve in the solvent. Therefore, the one having a low alkylation rate was reacted with the water-insoluble polymer. Thereafter, it is convenient to further add a predetermined amount of an acid chloride or acid anhydride of an alkyl bromide or a carboxylic acid to the reaction system to obtain a target alkylation ratio. In particular, when the substituent is an acyl group, the latter is preferably used because it is difficult to dissolve in a solvent.

The method for producing the adsorbent of the present invention includes a homogeneous reaction method in which a water-insoluble polymer and an N-substituted poly (alkyleneimine) are reacted in a solution, and a method in which a water-insoluble polymer is molded into a water-insoluble polymer. Any of the methods of heterogeneous reaction of contacting a substituted poly (alkyleneimine) solution can be employed.

An example of a method for producing an adsorbent by a homogeneous reaction is as follows. A corresponding polyamine is added to a solution of chloroacetamidomethylated polysulfone and the solution is added at 0 to 100 ° C.
It is easily manufactured by reacting at a temperature of The amount is not particularly limited, but is preferably at least 1 mole per haloacetamide methyl group in order to obtain a soluble polymer. In particular, in the case of a branched polyamine, it is preferable to use a large excess of the polyamine in order to obtain a soluble polymer.

When the reaction is carried out in a homogeneous system, tetrahydrofuran, dimethyl sulfoxide, N, N-dimethylformamide (DMF), N, N
N-dimethylacetamide, N-methylpyrrolidone and the like are preferably used. In addition, a method of surface-treating a molded product such as a membrane or a fiber can be employed. For this purpose, a solvent that does not dissolve polysulfone such as water, methanol, and ethanol but dissolves polyamine is preferably used.

As an example of a method for producing an adsorbent by a heterogeneous reaction, a molded article such as chloroacetamidomethylated polysulfone fiber or hollow fiber is immersed in an N-alkylated polyalkyleneimine solution in isopropanol,
It is easily manufactured by reacting at a temperature of 0 to 100 ° C.

It is practically preferable to coat the surface of a molded product such as nylon fiber with such an adsorbent, since a high-order molded product having a large surface area can be easily obtained. As a coating method, it is possible to coat easily by immersing the knitted fabric or woven fabric of nylon in a solution in which the adsorbent is dissolved in a low boiling point solvent such as methylene chloride or tetrahydrofuran, and then evaporating the solvent. Also,
A wet coating method in which a substance dissolved in a solvent such as N, N-dimethylformamide is placed in water or the like can also be used. The polymer of the molded article to be coated may be any polymer having good adhesiveness with the adsorbent of the present invention, such as polyamide, polyurethane, polyimide, polysulfone, polyvinyl chloride, and polyester, and the type thereof is not particularly limited. However, since amide polymers such as nylon and polyetherimide have particularly good adhesion,
It is preferably used.

When the adsorbent of the present invention is used for extracorporeal circulation,
It is preferable to use the anticoagulant treated with heparin, since the blood compatibility is improved and the adsorbability to LDL is improved. In particular, when the C / N ratio is high, the adsorbent becomes hydrophobic, but heparin treatment is preferred because it becomes hydrophilic and the adsorbing ability increases.

In the present invention, the specific adsorbent thus obtained is packed in a column and used. That is, such a column has a blood inflow portion for flowing blood into the adsorption portion filled with the adsorbent, and a blood outflow portion for flowing out the blood that has flowed into the adsorption portion. Any structure that does not cause drift is used without particular limitation.

[0029]

The present invention will be described more specifically with reference to the following examples.

Example 1 A mixed solution of 16 mL of nitrobenzene and 32 mL of sulfuric acid was diluted with 0
After cooling to ° C., 4.2 g of N-methylol-α-chloroacetamide was added and dissolved. This was dissolved in a 3 L nitrobenzene solution (300 g / 300 g / ml) of udel polysulfone (P3500, manufactured by Teijin Amoco Engineering Co., Ltd.) at 10 ° C.
To 3 L) with good stirring, and further stirred at room temperature for 3 hours. Thereafter, the reaction mixture was placed in a large excess of cold methanol to precipitate the polymer. The precipitate was thoroughly washed with methanol and then dried to obtain 300 g of α-chloroacetamidomethylated polysulfone (substitution ratio: 0.05; polymer A).

Polyethyleneimine (average molecular weight 1000
0: Wako Pure Chemical Industries, Ltd.) A solution of 20 g in 50 mL of DMF was added with 11.4 g of lauryl bromide, and the mixture was stirred at 50 ° C. for 5 hours. Then, 10 g of polymer A was added to 100 ml of DM.
The solution dissolved in F was added and heated at 50 ° C. for 5 hours.

[0032] 11.4 g of lauryl bromide was added to the reaction mixture, and the mixture was further stirred at 50 ° C for 8 hours, then added to isopropanol, and the precipitated polymer was collected by filtration.

The obtained polymer was dried under vacuum to obtain DM
F, reprecipitated with isopropanol, dried under vacuum, and N-alkylated polyalkyleneimine-bound polysulfone having a C / N ratio of 4.4 and an alkylation ratio of 20% (Example 1).
Adsorbent 1) was prepared.

A C / N ratio of 7.8 and an alkylation ratio of 4 were obtained in the same manner as described above, except that lauryl bromide was used in a double amount.
A 0% N-alkylated polyalkyleneimine linked polysulfone (Adsorbent 2 of Example 2) was prepared.

Adsorbents 1 and 2 of Examples 1 and 2
Was immersed in 250 ml of a solution of 3% concentration in tetrahydrofuran, and each 20 g of nylon 6 having a fiber diameter of 10 μm and 52 filaments was immersed. After 1 hour, the tube was taken out, drained, air-dried, and 21 g each (The knitted fabrics 1 and 2 of Examples 1 and 2) were obtained.

After 0.5 g of each of these knitted fabrics was subjected to autoclave sterilization, the adsorbability was evaluated (at 37 ° C. for 2 hours) using 15 ml of bovine serum containing endotoxin. The results shown in Table 1 were obtained.

[0037]

[Table 1]

However, in Comparative Example 1, uncoated nylon 6 was used.
It is a knitted fabric.

Further, knitted fabrics 1 and 2 of Examples 1 and 2
For, the adsorptivity of granulocytes was examined, and the results in Table 2 were obtained.

To a New Zealand White rabbit (3.5 kg in weight), LPS (E. coli O111: B4 dissolved at a concentration of 100 μg / mL) was administered at 5 μg / kg body weight via the ear vein, and 14 days later, the same amount was used. 2 days after administration of LPS, blood was collected from the ear artery,
It was used for the following evaluation.

After 100 mg of the adsorbent was placed in 2 ml of the above blood and shaken at 37 ° C. for 1 hour, the composition of the blood cells was examined by an automatic hematology analyzer, and the ratio of granulocytes to lymphocytes was determined by a flow cytometer. When the numbers of the blood cells were calculated, the results shown in Table 2 were obtained.

[0042]

[Table 2]

From Tables 1 and 2, the knitted fabric 1 of Examples 1 and 2 was obtained.
It can be seen that the column packed with Nos. And 2 has the ability to adsorb both endotoxin and granulocytes, whereas the column of Comparative Example 1 adsorbs granulocytes but does not adsorb endotoxin.

[0044]

According to the present invention, an adsorbent capable of simultaneously removing both toxins derived from bacteria in blood and inflammatory leukocytes, which had not been known before, can be provided stably with good reproducibility, and , Can quickly treat inflammatory diseases.

Continuation of the front page F term (reference) 4C077 AA11 BB03 CC06 EE01 HH03 KK27 LL02 LL05 NN14 NN15 PP13 PP15 4D017 AA11 BA20 CA13 CB01 DA02 DA03 EA01

Claims (11)

[Claims] 1. A column for treating inflammatory diseases, which is filled with an inflammatory substance adsorbent capable of removing both bacterial toxins and inflammatory leukocytes in blood. 2. The column for treating inflammatory diseases according to claim 1, wherein the inflammatory substance adsorbent is a water-insoluble polymer having an N-substituted poly (alkyleneimine) residue bonded to a side chain. 3. The method according to claim 2, wherein the N-substituted poly (alkyleneimine) residue has a basic nitrogen atom, and the ratio of the number of carbon atoms to the number of basic nitrogen atoms is 2.3 to 26. Column for treating inflammatory diseases. 4. The N-substituted poly (alkylimine) residue, wherein the N-substituent is an alkyl group or an acyl group having 4 or more carbon atoms, and the number of N-substituents is the same as the number of nitrogen atoms. 1
The column for treating inflammatory diseases according to claim 2 or 3, wherein the concentration is not less than 30%. 5. The poly (alkyleneimine) moiety of the N-substituted poly (alkyleneimine) residue has a degree of polymerization of 4 or more and 1
The column for treating an inflammatory disease according to any one of claims 2 to 4, which has a molecular weight of 0000 or less. 6. The column for treating an inflammatory disease according to claim 2, wherein the water-insoluble polymer is a polysulfone polymer having an amine binding group. 7. The column for treating an inflammatory disease according to claim 2, wherein the water-insoluble polymer is a poly (vinyl aromatic compound) having an amine-binding group. 8. The column for treating an inflammatory disease according to claim 2, wherein the water-insoluble polymer is a polyimide polymer. 9. The method according to claim 1, wherein the polysulfone-based polymer is poly (p-
7. The column for treating an inflammatory disease according to claim 6, which is phenylene ether sulfone). 10. The polysulfone-based polymer, wherein-｛(p
_{-C 6 H 4) -SO 2 -} (p-C 6 H 4) -O- (p-
_{C 6 H 4) -C (CH} 3) 2 - (p-C 6 H 4) -O}
The inflammatory response protein adsorbent according to claim 6 or 9, which is a polymer composed of a unit represented by-. 11. The adsorbent is at least one selected from a membrane, a fiber, a hollow fiber, and a granular material.
0. The column for treating an inflammatory disease according to any one of 0.