JPH1085329A - Material for removing or detoxificating polypeptide-based antibiotics - Google Patents

Abstract

PROBLEM TO BE SOLVED: To remove effectively antibiotics, above all, polypeptide-based antibiotics in blood, by using a substance with a functional group forming the hydrogen bonding ability, and more preferably, a substance with an urea or thiourea bond. SOLUTION: This detoxifying substance best suitable for removing polypeptide-based antibiotics, particularly vancomycin, has a functional group possible to form a hydrogen bond. As functional groups possible to form a hydrogen bond, such groups as hydroxyl, carboxyl, mercapto, acetyl, acyl, amide and so forth are used. These potential hydrogen bonding functional groups can also be bonded with a variety of substitution groups, and for instance, such compounds as aliphatic, alicyclic, aromatic and the like are used. At least one of amino groups uses a secondary or tertiary amino group so as to have saccharinity contain hydroxyl group.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a material for removing or detoxifying a polypeptide antibiotic. In particular, it is suitably used as a purification column for removing polypeptide antibiotics present in human blood or the like, or as a drug that loses toxicity (detoxification) by binding to polypeptide antibiotics.

[0002]

2. Description of the Related Art Among attempts to develop antibacterial agents effective against various infectious diseases, polypeptide antibiotics such as vancomycin, polymyxin B, teicoplanin, colistin, and vantramin have excellent bactericidal properties. Nevertheless, its high toxicity limits its use. Among them, vancomycin is specific to aerobic and anaerobic Gram-positive bacteria such as Staphylococcus, including methicillin-resistant Staphylococcus aureus (MRSA); It is known that it exhibits a strong antibacterial and bactericidal action, and its mechanism of action is to inhibit the bond between the murein cross-linking enzyme and the substrate by forming a hydrogen bond with D-alanyl-D-alanine, which is a substrate of mulein. And it is known to act as a cell wall peptidoglycan synthesis inhibitor of Gram-positive bacteria.

For clinical use, since its intravenous administration for Gram-positive bacterial infections in the United States in 1958 has been approved, it is currently approved for use in more than 100 countries worldwide, and is especially known as a specific agent for MRSA. .

[0004] However, it has been known that side effects such as hepatic dysfunction, hearing impairment, renal dysfunction and hypersensitivity occur in clinical use of vancomycin. Because vancomycin undergoes little metabolism in vivo and its main excretion route is the kidney, it is particularly suitable for administration to patients with impaired renal function, such as the elderly, renal dysfunction patients, dialysis patients, and neonates. Be careful.

[0005] Many of these side effects are said to appear depending on the concentration of vancomycin in the blood, but at this stage, even if the side effects of vancomycin appear once, there is no means for rapidly reducing the blood concentration of vancomycin. , High
There is only hemodialysis using performance membrane (HPM). It is said that about 50% of antibiotics in blood are removed by using HPM in hemodialysis. However, dialysis removal is time-consuming, and the dialysis system is complicated. Also, the use of dialysis membranes that are permeable to large molecular weights, such as HPM, often suffers from the loss of useful plasma proteins such as albumin. Also,
Polymyxin B, which is the same polypeptide antibacterial drug, has excellent antibacterial activity against Gram-negative bacteria, but its nephrotoxicity makes it impossible to administer it in the blood, and its oral administration is restricted.

[0006]

As described above, a simpler and more efficient means for removing antibiotics in blood, especially polypeptide antibiotics, which is different from hemodialysis, is desired. Therefore, an object of the present invention is to provide a material having high selectivity or affinity for polypeptide antibiotics.

[0007]

Means for Solving the Problems As a result of intensive studies, the present inventors have found that vancomycin, an antibacterial agent, is contained in a material having a functional group capable of forming a hydrogen bond, more preferably a material having a urea bond or a thiourea bond. The present inventors have found that they have high affinity with the first polypeptide antibiotics, and have led to the present invention. That is, the present invention has the following configuration.

(1) A material for removing or detoxifying a polypeptide antibiotic, which has a functional group capable of forming a hydrogen bond.

(2) A body fluid purification column using a material having an affinity for the polypeptide antibiotic described in 1 above.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, a functional group capable of forming a hydrogen bond (hereinafter referred to as a hydrogen bond-forming functional group) includes a hydroxyl group, a carboxyl group, a mercapto group, an acetyl group, an acyl group, an amide group. And the like, and particularly, an amino group, a urea bond, a thiourea bond and the like are preferably used. Various substituents can be bonded to these hydrogen bond-forming functional groups, and examples thereof include aliphatic compounds, alicyclic compounds, aromatic compounds, and derivatives of the above substituents. Specifically, hexyl group, octyl group, dodecyl group, cyclohexyl group, cyclopentyl group, phenyl group, diphenylmethyl group, naphthyl group, aminohexyl group, monomethylaminohexyl group, dimethylaminohexyl group, aminooctyl group, aminododecyl Groups, tolyl group, chlorophenyl group, nitrophenyl group, aminodiphenylmethyl group and the like. Further, as a substituent to be bound, monosaccharides such as glucose, glucosamine, galactosamine, maltose, cellobiose, sucrose, agarose, cellulose, chitin, chitosan and the like, and saccharides such as oligosaccharides and polysaccharides or derivatives thereof are also preferably used. Among them, in the present invention, it is most preferable to have both an aromatic compound and a hydroxyl group or an amino group as a substituent of a urea bond or a thiourea bond.

The material of the present invention may be any of a monomer, an oligomer and a polymer, and the above-mentioned substituent or a polymer of a part thereof is also included in the material of the present invention. That is, as the functional group or a part thereof of the material of the present invention, nylon, polymethyl methacrylate, polysulfone, polystyrene, polyethylene, polyvinyl alcohol, polytetrafluoroethylene, polyallylamine, polyvinylamine, synthetic polymers such as polyethyleneimine, A repeating unit such as a natural polymer containing cellulose, collagen, chitin, chitosan and a derivative thereof is preferably used. Further, a synthetic polymer or a natural polymer into which an amino group or a hydroxyl group which is a hydrogen bond-forming group or a urea bond or a thiourea bond is introduced may be homopolymerized, or may be copolymerized or blended. Further, a support obtained by coating a support made of an inorganic material such as metal, ceramic, glass or the like with a suitable polymer is also preferably used.

The material of the present invention can be synthesized by a generally known method. For example, among the functional groups capable of forming hydrogen bonds, as a method for introducing a urea bond or a thiourea bond into an aliphatic compound or an aromatic compound, a method of reacting an isocyanate derivative or an isothiocyanate derivative with an amine is used. Can be. Although the amount ratio of the reagents can be arbitrarily selected, it is preferable that the amine is 1 mol per 0.1 to 5 mol of the isocyanate derivative or isothiocyanate derivative. When a urea bond or a thiourea bond is introduced into a saccharide, a saccharide having an amino group such as chitosan or glucosamine can be reacted with an isocyanate derivative or an isothiocyanate derivative. In the case of a saccharide having no amino group such as cellulose, the hydroxyl group of the saccharide is activated using epichlorohydrin, tresyl chloride, etc., and then reacted with ammonia or diaminoethane to introduce an amino group. Utilizing this amino group, a urea bond or a thiourea bond can be introduced into the saccharide.

When the material of the present invention is an oligomer or a polymer, for example, an oligomer or a polymer having an active ester group of a carboxylic acid such as an isocyanate group, a carboxyl group or a succinimide group may be added to an amino or urea derivative or a thiourea derivative. A method of reacting a group is preferably used. As the amino group used for the reaction, the amino group at the terminal of the urea bond or the thiourea bond is low in reactivity, and therefore, an amino group present at other sites is preferable. Usually, 1 to 5 moles of amino groups are preferably used for 1 mole of active ester groups contained in the oligomer or polymer. Oligomers having a primary amino group, polymers, or oligomers having an amino group introduced by diaminoethane or the like, phenyl isocyanate, p-tolyl isocyanate, p-chlorophenyl isocyanate, 1-naphthyl isocyanate, phenylisothiocyanate, Reaction of an organic isocyanate such as chlorophenyl isothiocyanate or an organic isothiocyanate is also a preferable method. Usually, 1 to 5 mol of organic isocyanate is preferably used for 1 mol of amino group contained in the oligomer or polymer. Functional groups such as an active ester group of a carboxylic acid such as an isocyanate group, a carboxyl group, and a succinimide group and an amino group can be introduced into an oligomer or a polymer as needed.

All of the above reactions are typically carried out at a reaction temperature of 0 to 150 ° C. and a reaction time of 0.1 to 24 hours, but are not limited thereto. Although a reaction solvent is not always necessary, the reaction is generally performed in the presence of a solvent. Solvents that can be used include aliphatic hydrocarbons such as methanol, ethanol, isopropyl alcohol, n-butanol, hexane, acetone, N, N dimethylformamide and dimethyl sulfoxide; halogenated hydrocarbons such as benzene, toluene and xylene; Ethers such as diethyl ether, tetrahydrofuran, and dioxane; After completion of the reaction, the reaction solution
It can be purified by an operation such as column chromatography and recrystallization after usual post-treatment such as filtration and concentration. In the case of a water-insoluble material, washing with a glass filter or the like is also a preferable method.

Since the material of the present invention has an affinity for polypeptide antibiotics such as vancomycin, vancomycin can be removed from body fluids such as blood by adsorbing vancomycin, for example. In addition, the use of medicine as an antidote is also possible. In particular, those insoluble in water are preferably used as vancomycin removal columns, analytical / diagnostic measurement materials, and the like. Also,
A water-soluble material can also be used as a vancomycin removal column, an analytical / diagnostic measurement material, etc. by immobilizing it on a water-insoluble material by a covalent bond. For example,
Utilizing a part of the functional group capable of forming a hydrogen bond, it can be immobilized on a synthetic polymer such as polysulfone or a natural polymer such as cross-linked chitosan and made insoluble in water. The shape is not particularly limited, but when used as a column, beads, fibers, hollow fibers, woven fabric, etc. are preferable, and when used for analytical / diagnostic measurement, beads, fibers, etc.
Plates, tubes and the like are preferred. For example, those obtained by introducing a functional group capable of forming a hydrogen bond into crosslinked chitosan beads are preferably used as vancomycin removal columns or analytical / diagnostic measurement materials.

Hereinafter, the present invention will be described in detail with reference to examples, but the present invention is not limited to the examples.

[0017]

【Example】

Example 1 Urea binding to chitosan beads and introduction of polyethyleneimine and vancomycin adsorption removal test using the beads Chitosan beads having the following structural formula [I] and having a particle diameter of 0.1 mm (“Chitopearl” manufactured by Fujibo Co., Ltd.) AL-01)
12 ml (sedimentation volume, dry weight 1.0 g) to 20 m
The mixture was stirred in 1 l of N, N-dimethylformamide (abbreviated as DMF), and the beads and the solution were separated by a glass filter. This operation was repeated once for 5 minutes for 20 times to completely replace the water content with DMF.

The beads were gradually added to 100 ml of DMF in which 1 g of p-chlorophenyl isocyanate was dissolved, and reacted at room temperature for 1 hour with stirring. Thereafter, the beads and the solution were separated using a glass filter, and the beads were washed by stirring them in 20 ml of DMF for 5 minutes. This washing operation was repeated 20 times to completely remove unreacted p-chlorophenyl isocyanate. Next, a washing operation with distilled water was performed in the same manner, and DMF was replaced with distilled water to obtain chitosan beads having the following structural formula [II].

Further, beads 12 in which the solvent was replaced with DMF were used.
was suspended in 90 ml of DMF, and while stirring, 10 ml of a DMF solution in which 0.5 g of epichlorohydrin was dissolved.
Was gradually added dropwise. With stirring, the reaction was further carried out at room temperature for 3 hours. Thereafter, the beads were washed 20 times in the same manner as described above using 20 ml of DMF using a glass filter. The beads are treated with polyethyleneimine (average molecular weight:
600) 9 g was dissolved in dissolved DMF and reacted at room temperature for 3 hours. Thereafter, the beads and the solution were separated using a glass filter, and the beads were washed by stirring them in 20 ml of DMF for 5 minutes. This washing operation was repeated 20 times to completely remove unreacted p-chlorophenyl isocyanate. Subsequently, washing operation with distilled water was performed in the same manner, and DMF was replaced with distilled water to obtain chitosan beads having the following structural formula [III].

The modified chitosan beads ([II], [III
]) And unmodified chitosan beads ([I]) as a control were used to remove vancomycin by adsorption in rabbit plasma. The initial concentration of vancomycin is 50
μg / ml, 120 ° C.
1 ml of the above chitosan beads after autoclaving for 20 minutes was added and shaken at 37 ° C. for 60 minutes. Table 1 shows the results of measurement of vancomycin in rabbit plasma after the reaction for 60 minutes by fluorescence polarization immunoassay. As shown by these results, the introduction of a urea bond imparted vancomycin adsorption ability to chitosan beads.

[0021]

Embedded image In the structural formula [III], n, m, and l each represent an integer of 0 or more.

[0022]

[Table 1] Example 2 Preparation of Polystyrene Fiber Having a Urea Derivative Having a Hydroxyl Group in the Side Chain From a 50 weight ratio of a sea component (a mixture of 46 weight ratio of polystyrene and 4 weight ratio of polypropylene) and a 50 weight ratio of an island component (polypropylene). U.S. Patent 4,661,260
50 g of N-methylol-α-chloroacetamide, 400 g of nitrobenzene, 400 g of 98 were added to the sea-island composite fiber (thickness: 2.6 denier, number of islands: 16).
Reaction with a mixed solution of 0.85 g of paraformaldehyde and 20% sulfuric acid at 20 ° C. for 1 hour. Then, the fiber was washed with nitrobenzene and put into water to stop the reaction. The chloroacetamidomethylated crosslinked polystyrene fibers (AMPSt) were then washed again with warm water.
(Abbreviated as fiber).

10 g of diaminohydroxypropane (abbreviated as DAHP) was dissolved in 500 ml of dimethyl sulfoxide (abbreviated as DMSO) DMSO. Also, 16 g of tetraethylenepentamine (TEPA) was added to 500 ml of DMF.
Was dissolved. 20 g of AMPS was added to each solution.
t-fiber (corresponding to a chloro content of 20 mmol) was added with stirring. The reaction was performed at 25 ° C. for 6 hours. Then AMPSt
On a glass filter with 500 ml of DMSO followed by D
Washed with 50 ml of MF. After washing, 1 g of AMPSt was added to AMPSt reacted with DAHP in a solution of 0.30 g of p-chlorophenylisocyanate in 50 ml of DMF. The reaction was performed at 25 ° C. for 1 hour. Then, 200 ml of DMF on a glass filter
And washed with 500 ml of distilled water. The obtained compound was designated as (a). In addition, AMP reacted with TEPA
St was washed on a glass filter with 200 ml DMF and 500 ml distilled water. The obtained compound was designated as (d).

The adsorption removal test of vancomycin using the modified polystyrene fiber (compound (a)) prepared as described above was performed in the same manner as in Example 1. The initial concentration of vancomycin was 50 μg / ml, and 1 g of modified AMPSt was added to 10 ml of plasma, and shaken at 37 ° C. for 60 minutes. All modified AMPSt fibers were used after autoclaving at 121 ° C. for 20 minutes. The result of having measured vancomycin in rabbit plasma after reaction for 60 minutes by the fluorescence polarization immunoassay is shown.

As a control, p-chlorobenzoic acid chloride was reacted under the same conditions in place of p-chlorophenyl isocyanate, and the AMPSt fiber compound (b) and DAH in which an amide bond was introduced instead of a urea bond.
AM obtained by reacting p-chlorophenylisocyanate after reacting diaminopropane (abbreviated as DAP) instead of P
The PSt fiber compound (c) was used.

As shown by these results, the polystyrene fiber having no urea bond (b) does not have the ability to adsorb vancomycin, indicating that the introduction of the urea bond exhibits the ability to adsorb vancomycin. In addition, it was revealed that vancomycin adsorption ability was enhanced by introducing a hydroxyl group.

[0027]

[Table 2]

[0028]

Industrial Applicability According to the present invention, it is a material having excellent adsorption to polypeptide antibiotics, excellent in controlling the concentration of the drug in blood, and effective in alleviating symptoms of drug intoxication. Material can be provided.

Claims (12)

[Claims] 1. A material for removing or detoxifying a polypeptide antibiotic, which has a functional group capable of forming a hydrogen bond. 2. The method according to claim 1, wherein at least one of the functional groups capable of forming a hydrogen bond has a structure having a urea bond or a thiourea bond. Or materials for detoxification. 3. The material for removing or detoxifying a polypeptide antibiotic according to claim 1, wherein the material has an aromatic ring. 4. The material for removing or detoxifying a polypeptide antibiotic according to claim 1, which has a hydroxyl group or an amino group. 5. The material for removing or detoxifying a polypeptide antibiotic according to claim 1, wherein at least one of the functional groups capable of forming a hydrogen bond is an amino group. 6. An amino group wherein at least one is 2
6. The material for removing or detoxifying a polypeptide antibiotic according to claim 5, wherein the material is a tertiary or tertiary amino group. 7. A compound having a hydroxyl group.
Material for removing or detoxifying the described polypeptide antibiotics 8. The material for removing or detoxifying a polypeptide antibiotic according to claim 7, wherein the hydroxyl group is contained in a saccharide. 9. The material for removing or detoxifying a polypeptide antibiotic according to claim 8, wherein the carbohydrate comprises at least one selected from chitosan, cellulose and derivatives thereof. 10. The material for removing or detoxifying a polypeptide antibiotic according to claim 1, wherein the polypeptide antibiotic is vancomycin. 11. The material for removing or detoxifying a polypeptide antibiotic according to claim 1, wherein said material is insoluble in water. 12. A body fluid purification column using the material for removing or detoxifying a polypeptide antibiotic according to any one of claims 1 to 11.