JP2002017850A - Material for treating cardiac failure and blood cleaning column - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a material which prevents the degradation in a cardiac function by detoxicating on neutralizing the cardiac function suppressing material existing in blood or restoring the degraded cardiac function and to provide a blood cleaning column for removing the cardiac function suppressing material, more particularly the cardiac function suppressing material derived from bacteria by using the material described above. SOLUTION: The material for detoxicating or removing the cardiac function suppressing material is provided by the material which has an antibody or peptide or urea bond or thiourea bond or amide bond having affinity to the cardiac function suppressing material.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a material for treating heart failure and a blood purification column.

[0002]

2. Description of the Related Art Diseases such as myocardial infarction, myocarditis, cardiomyopathy, valvular disease, cardiac tamponade and endocarditis, as well as the influx of toxins caused by cardiotoxic drugs and infections often cause the heart pump function to deteriorate. However, a state in which a sufficient amount of blood cannot be ejected for the metabolic demand of living tissue, that is, heart failure is caused. Furthermore, when peripheral vascular resistance is reduced due to infection or the like, a sharp decrease in blood pressure occurs, and a complication of heart failure results in a shock state.

[0003] In the treatment of such heart failure, an inotropic agent for enhancing the function of the heart is first administered. As a cardiotonic agent, dopamine or dobutamine which is a β-receptor stimulant that enhances the contractile force of the heart and increases the heart rate is used. Further, when blood pressure is lowered due to dilation of peripheral blood vessels, epinephrine or norepinephrine, which is an α receptor stimulant having a vasoconstrictive action, may be used. These sympathetic stimulants have a short time to act and have a remarkable effect of restoring the contractile force of the myocardium, and are currently widely used.

[0004] However, sympathetic stimulants have no therapeutic effect on the underlying disease, and cardiac function declines again when drug administration is stopped in a state where cardiac function-suppressing factors such as influx of toxin are continuously acting. Furthermore, administration of a large amount of a sympathetic stimulant causes side effects such as arrhythmia, gastrointestinal disorders, and peripheral vascular necrosis.

[0005]

SUMMARY OF THE INVENTION The present inventors have focused on a substance that suppresses the function of the heart, which is a cause of heart failure, which was not considered in these prior arts, and identified the substance. The present inventors have conducted intensive studies from a new viewpoint of preparing a material which has an affinity for the inhibitor and removes the toxicity from the body by detoxifying or adsorbing the toxicity.

[0006]

That is, the present invention is a material for treating heart failure, which removes or detoxifies a substance inhibiting cardiac function in blood.

[0007] The present invention is also a blood purification column filled with the above-mentioned material for treating heart failure.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION It is important for the material for treating heart failure of the present invention to detoxify or remove a cardiac function inhibitor. Examples of the cardiac function inhibitor include chemicals, pharmaceuticals, endogenous proteins, exogenous toxin proteins, and the like.In the present invention, in particular, attention is paid to bacterial toxins as exogenous substances, The effect of suppressing odor was examined. Examples of bacteria to which the toxin is attributed include Escherichia coli, Pseudomonas aeruginosa, Gram-negative bacteria such as Klebsiella pneumoniae or Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus, and Gram-positive bacteria such as Bacillus subtilis. And superantigen-producing bacteria such as Staphylococcus aureus and group A streptococci were examined.

[0009] The superantigen, unlike conventional antigens, is converted into a major histocompatibility class II protein (hereinafter abbreviated as MHC class II) on the antigen-presenting cells without undergoing a processing step in the antigen-presenting cells. A compound that binds, and further stimulates T cells having a specific V region by forming a complex with this MHC class II, and abnormally activates the immune system.
It is said to cause a decrease in blood pressure, vomiting during food poisoning, or induction of autoimmune diseases. As a super antigen,
Staphylococcal enterotoxin A, B, C, D, E, H, which is a Staphylococcus aureus exotoxin, or toxic shock syndrome toxin-1 (hereinafter abbreviated as TSST-1), or Streptococcus fever, which is a group A streptococcal exotoxin. More than a dozen types have been identified, including sex exotoxin A and certain viral and plant proteins, but may be identified in the future.

In the present invention, as a material having an affinity for a cardiac function-suppressing substance, a low molecular structure such as an antibody or a peptide specifically recognizing these substances or a functional group of a chemically synthesized affinity chromatograph. There are various possibilities, and there is no particular limitation.

An antibody that specifically recognizes a cardiac function-suppressing substance can be prepared by a conventional method (Biochemical Experimental Method 1).
1 "Enzyme immunoassay" Tijssen
Author, translated by Eiji Ishikawa, 1989, Tokyo Doujinshi). In addition, a peptide that specifically suppresses a cardiac function-suppressing substance can be prepared by a known method (Sato. A).
Biochemistry, Vol. 35, No. 10441-
10447, 1996). Preferably, a chemically synthesized low molecular structure which is inexpensive and easily sterilized is used, and more preferably, a structure having a urea bond, a thiourea bond or an amide bond is used. The structure following the urea bond, thiourea bond or amide bond is not particularly limited, and an aliphatic compound such as propane, hexane, octane, and dodecane and an alicyclic compound such as cyclohexane and cyclopentane can be used. Considering the high affinity, aromatic compounds such as benzene, naphthalene, and anthracene are more preferably used. Derivatives such as bromoheptane, chlorocyclohexane, methylbenzene, chlorobenzene, nitrobenzene, diphenylmethane, and chloronaphthalene are also suitably used. A structure having an amino group, a hydroxyl group or a carboxyl group is used. For example, as a structure having an amino group,
Aminohexane, monomethylaminohexane, dimethylaminohexane, aminooctane, aminododecane, aminodiphenylmethane, 1- (3-aminopropyl) imidazole, 3-amino-1-propene, aminopyridine, aminobenzenesulfonic acid, tris (2- Aminoethyl) amine, diaminoethane, diethylenetriamine, triethylenetetramine, tetraethylenepentamine, dipropylenetriamine, polyethyleneimine,
N'-methyl-2,2'diaminodiethylamine, N-
A compound having a plurality of amino groups (sometimes referred to as polyamine) such as acetylethylenediamine, 1,2-bis (2-aminoethoxyethane), or the like is used. Examples of the structure having a hydroxyl group include hydroxypropane, 2-ethanolamine, 1,3 diamino-2-hydroxypropane, hydroxybutanone, hydroxybutyric acid,
Hydroxypyridine, glucose, glucosamine,
Monosaccharides such as galactosamine, maltose, cellobiose, sucrose, agarose, cellulose, chitin, chitosan, etc., saccharides such as oligosaccharides and polysaccharides, and derivatives thereof can be used. Further, as the structure having a carboxyl group, β-alanine, n-caproic acid, isobutyric acid, γ-amino-β-hydroxybutyric acid, and the like can be used. Most preferably, the material of the present invention can have an aromatic compound and an amino group or a hydroxyl group as a structure following a urea bond, a thiourea bond or an amide bond.

Further, polyurea, polythiourea and polyamide having a plurality of urea bonds, thiourea bonds and amide bonds in the molecular structure can be used as the material of the present invention. Also in this case, any of the above structures can be used as a structure following the urea bond, thiourea bond, or amide bond, but most preferably, both a hydroxyl group, an amino group or a carboxyl group and an aromatic compound are used. Can be.

Since the material of the present invention may be any of a monomer, an oligomer and a polymer, the material of the present invention includes those having the above structure or a part thereof polymerized. That is, as the above structure or a part thereof, synthetic polymers such as nylon, polymethyl methacrylate, polysulfone, polystyrene, polyethylene, polyvinyl alcohol, and polytetrafluoroethylene, and natural materials containing cellulose, collagen, chitin, chitosan, and derivatives thereof. A polymer or the like is preferably used. That is,
It is preferable to introduce a urea bond and / or a thiourea bond group and / or an amide bond into these homopolymerized, copolymerized or blended synthetic polymers and natural polymers. Further, a material obtained by coating an inorganic material such as metal, ceramics, and glass with a suitable polymer is also preferably used.

The material of the present invention can be synthesized by a generally known method. For example, when a urea bond or a thiourea bond is introduced into an aliphatic compound or an aromatic compound, a method of reacting an isocyanate compound or an isothiocyanate compound with an amino compound can be used. When an amide group is introduced into an aliphatic compound or an aromatic compound, for example, a method of reacting an acid, an acid chloride or an acid anhydride with an amino compound can be used. The mixing ratio of the amino compound and the isocyanate compound, the isothiocyanate compound, the acid, the acid chloride or the acid anhydride can be arbitrarily selected, and is usually 1 mol per 1 mol of the isocyanate compound, the isothiocyanate compound, the acid, the acid chloride or the acid anhydride. 0.1 to 10 mol of the amino compound is preferably used. As the isocyanate compound or isothiocyanate compound, for example, ethyl isocyanate, stearyl isocyanate, n-butyl isocyanate, iso-butyl isocyanate, n-propyl isocyanate, methyl isothiocyanate, ethyl isothiocyanate, n-butyl isothiocyanate, benzyl isothiocyanate, hexyl isocyanate Any of aliphatic isocyanate compounds such as methylene diisocyanate, cyclohexyl isocyanate, cyclohexyl isothiocyanate, and cyclohexyl diisocyanate or isothiocyanate compounds can be used, but phenyl isocyanate, chlorophenyl isocyanate, fluorophenyl isocyanate, bromophenyl isocyanate, bromophenyl isocyanate, Isocyanate, tolyl isocyanate, methoxyphenyl isocyanate, 1-naphthyl isocyanate, 4,4'diphenylmethane diisocyanate, 3,3 ', 5,5'tetraethyl 4,4'diisocyanatodiphenylmethane, phenylisothiocyanate, chlorophenylisothiocyanate, fluorophenyl Aromatic isocyanate compounds such as isothiocyanate, nitrophenyl isothiocyanate, tolyl isothiocyanate, methoxyphenyl isothiocyanate, 1-naphthyl isothiocyanate and the like, or isothiocyanate compounds are used. As the acid chloride, for example, any one of aliphatic acid chlorides such as isovaleryl chloride, stearoyl chloride, cyclohexanecarbonyl chloride, and 6-chloronicotinic acid chloride can be used. Aromatic acid chlorides such as 4-dichlorobenzoyl chloride, nitrobenzoyl chloride, 4-chlorobenzoyl chloride, 4-toluoyl chloride and benzo- [b] thiophen-2-carbonyl chloride can be used. As the acid anhydride, for example, acetic anhydride, succinic anhydride, phthalic anhydride, benzoic anhydride and the like can be preferably used. Further, as the amino group of the amino compound used in the present invention, a primary amino group,
Any of a secondary amino group and a tertiary amino group may be used. Examples of the amino compound include ammonia, sec-octylamine, 1- (3-aminopropyl) imidazole,
-Amino-1-propene, aminopyridine, aminobenzenesulfonic acid, tris (2-aminoethyl) amine and the like can be preferably used. Further, a polyamino compound or an amino compound having a hydroxyl group or a carboxyl group which can introduce a urea bond, a thiourea bond or an amide bond can also be preferably used. Examples of the polyamino compound include diaminoethane, diethylenetriamine, triethylenetetramine, tetraethylenepentamine, dipropylenetriamine, and N′-methyl-.
2,2 ′ diaminodiethylamine, polyethyleneimine,
Any of N-acetylethylenediamine, 1,2-bis (2-aminoethoxy) ethane and the like can be used. Examples of the amino compound having a hydroxyl group include 2-ethanolamine, 3-propanolamine, 6-hexanolamine, 1,3 diamino-2-hydroxypropane, 2- (2-aminoethoxy) ethanol, and 2- (2-aminoethyl Amino) Ethanol, glucamine, aliphatic amines such as N-methyl 1,3-diaminopropanol, or aromatic amines such as 4-aminophenol, diaminophenol, aminohydroxypyrimidine, diaminohydroxypyrimidine, diaminohydroxypyrazole, or serine ,
Amino acids such as tyrosine are used. Epichlorohydrin and amino compounds, or 1,3-dibromo-
By reacting 2-hydroxypropane, an amino compound having a hydroxyl group can be synthesized from a compound having only a hydroxyl group or a compound having only an amino group. In addition, when a urea bond, a thiourea bond or an amide bond is introduced into a saccharide, the same method as described above can be used. That is, in the case of a carbohydrate having an amino group such as chitosan or glucosamine, an isocyanate compound as described above, an isothiocyanate compound,
An acid, acid chloride or acid anhydride can be reacted. 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, diaminoethane, etc. to introduce an amino group. Utilizing this amino group, a urea bond, a thiourea bond, an amide bond or the like can be introduced into the saccharide. As the amino compound having a carboxyl group, for example, β-alanine, 4-amino-n-butyric acid, γ-amino-β-hydroxy-n-butyric acid, 6-amino-n-caproic acid and the like can be used.

Further, when the material of the present invention is an oligomer or a polymer, for example, a method of reacting an amino group with 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 is preferable. Used. Oligomers, polymers or ammonia having amino groups,
Diaminoethane, 1,3 diaminopropane, 1,3 diamino-2
-Hydroxypropane, 1,2-bis (2-aminoethoxy) ethane, tris (2-aminoethyl) amine, 2-
It is also a preferable method to react an oligomer or polymer having an amino group introduced with (2-aminoethylamino) ethanol or the like with the above-mentioned isocyanate compound, isothiocyanate compound, acid, acid chloride or acid anhydride. Further, it is preferable to control the reaction time, the reaction temperature, the mixing ratio, and the like, and to use a protecting group so that the acid chloride and the isocyanate compound do not react with groups other than the amino group. Amino group, isocyanate group, active ester group of carboxylic acid such as carboxyl group or succinimide group, acid chloride group, functional group such as acid anhydride group,
It can be introduced into an oligomer or a polymer as needed.

Further, when the material of the present invention is polyurea or polythiourea, for example, a method of reacting a polyisocyanate compound or a polyisothiocyanate compound with a polyamino compound can be used. Normal,
The amount of the reagent is preferably 0.1 to 10 mol of the polyamine per 1 mol of the polyisocyanate compound or the polyisothiocyanate compound. As polyisocyanate compounds or polyisothiocyanate compounds, hexamethylene diisocyanate, cyclohexyl diisocyanate, tolylene diisocyanate, 4,4 'diphenylmethane diisocyanate, 3,3', 5,5 'tetraethyl 4,4' diisocyanato diphenylmethane, xylene diisocyanate, Methylene bis (4-phenylisothiocyanate) and the like are preferably used. As polyamino compounds, diaminoethane, diaminopropane, 1,
3 diamino-2-hydroxypropane, N'-methyl 1,3-
Diamino-2-propanol, diaminophenol, N,
N'-diaminopiperazine, diethylenetriamine, triethylenetetramine, tetraethylenepentamine, polyethyleneimine, dipropylenetriamine, N'-methyl-2,2'-diaminodiethylamine and the like can be preferably used. Further, when the material of the present invention is a polyamide, for example, a method of polycondensing a polycarboxylic acid and a polyamine can be used. In addition, in any of polyurea, polythiourea and polyamide, polyurea, polythiourea and polythiourea are finally introduced by sequentially introducing each functional group one by one without using polyisocyanate, polyisothiocyanate, polycarboxylic acid and the like. urea,
A method for obtaining a polyamide is also preferably performed.

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. Although a reaction solvent is not always necessary, the reaction is generally performed in the presence of a solvent. Examples of usable solvents include aliphatic hydrocarbons such as methanol, ethanol, isopropyl alcohol, n-butanol, hexane, acetone, N, N dimethylformamide and dimethyl sulfoxide, and aromatic hydrocarbons such as benzene, toluene and xylene. And halogenated hydrocarbons such as dichloromethane, chloroform and chlorobenzene, and ethers such as diethyl ether, tetrahydrofuran and dioxane. After completion of the reaction, the reaction solution can be purified by an operation such as column chromatography or recrystallization, if necessary, after usual post-treatments 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.

Among the materials of the present invention, those which are insoluble in water are preferably used as columns for removing cardiac function-inhibiting substances.
The shape is not particularly limited, but when used as a column, beads, fibers, hollow fibers, yarn bundles, yarns,
Nets, knitted fabrics, woven fabrics and the like are preferred. In addition, this material can be used not only alone, but also immobilized on an appropriate material, or mixed with another material and used as one column or coating material. Operations such as fixing or mixing may be performed before processing into the above-mentioned shape, or may be performed after processing. When a column using the material of the present invention is used as a column for extracorporeal circulation, blood extracted outside the body may be directly passed through the column, or may be used in combination with a plasma separation membrane or the like.

[0019]

EXAMPLES Example 1 Preparation of a Material Having Affinity with Cardiac Inhibitors 50 weight ratio of sea component (a mixture of 46 weight ratio of polystyrene and 4 weight ratio of polypropylene) and 50 weight ratio of island component (polypropylene) US Pat. No. 4,661,260 consisting of
Of the sea-island composite fiber (thickness: 2.6 denier, number of islands: 16) described in Example 1 of Example 1 was treated with 50 g of N-methylol-α-
Chloroacetamide, 400 g nitrobenzene, 40
The mixture was reacted with a mixed solution of 0 g of 98% sulfuric acid and 0.85 g of paraformaldehyde at 20 ° C. for 1 hour. Thereafter, the fiber was washed with nitrobenzene, the reaction was stopped with methanol, and then washed with methanol to obtain an α-chloroacetamidomethylated crosslinked polystyrene fiber (hereinafter abbreviated as AMPSt fiber).

Next, 6.3 of tetraethylenepentamine
g and tert-butylamine (7.2 g) were dissolved in 500 ml of N, N dimethylformamide (hereinafter abbreviated as DMF), and 20 g of AMPSt fiber was added with stirring. The reaction was performed at 30 ° C. for 3 hours, and then the reacted fiber was washed on a glass filter using 500 ml of DMF, and the fiber was added to a solution of 2.3 g of 4-chlorophenylisocyanate in 500 ml of DMF, and the reaction was performed. For 1 hour at 25 ° C. Thereafter, the resultant was washed with 1000 ml of DMF on a glass filter, and further washed with 1000 ml of distilled water to obtain AMPSt fiber (abbreviated as UAMP fiber) into which a urea bond was introduced as a material for treating heart failure of the present invention.

Here, using the produced UAMP fiber,
Toxic shock syndrome toxin-1 (TSS), a superantigen derived from bacteria (Staphylococcus aureus)
Abbreviated as T-1. (Toxin Technology Co., Ltd.). 0.1 M sodium phosphate buffer (pH 7.4) containing 0.15 M sodium chloride and 5 mg / ml bovine serum albumin (fraction V) (manufactured by Seikagaku Corporation) so that TSST-1 has a concentration of 1 ng / ml. It was dissolved in the solution to obtain a solution to be absorbed. 0.1 g of UAMP was added to 1 ml of the solution to be adsorbed, and the adsorbing reaction was carried out while rotating and stirring at 37 ° C. for 2 hours. The TSST-1 concentration before and after adsorption was measured. (Table 1)

[0022]

[Table 1]

Thus, it was shown that fibers having a high adsorption rate to TSST-1 were produced.

Example 2 A pig (25 kg; female) was anesthetized, endotracheally intubated, and controlled with an oxygen concentration of 30% using an artificial respirator. Anesthesia was continued continuously with gas anesthesia. In addition, physiological saline was used as an infusion at 3 ml / k.
It was continuously administered intravenously at g / hr. A Swan-Ganz catheter was inserted through the jugular vein.

Toxic shock syndrome toxin-1 (TSST-1) as a bacterial toxin was 5 μg / kg / kg.
It was administered intravenously over 1 hour at a rate of hr. Then 4
Time cardiac function was measured with a Swan-Ganz catheter. The work function of the left ventricle was used as the cardiac function.

The following three types of treatment were performed on the toxin-administered pig. (1) Administration of polyclonal anti-TSST-1 antibody. (2) Administration of TSST-1 neutralizing peptide.
(3) Blood purification by TSST-1 adsorbing material.

In (1), an anti-TSST-1 antibody purified by a TSST-1 immobilized column from an antiserum collected from a rabbit immunized with TSST-1 was injected 1 m 5 minutes before administration of the toxin.
After intravenous injection of g into pigs, administration of TSST-1 is started under the above conditions.

In (2), the anti-TSST-administered in (1)
Amino acid sequence DRSYLSFIHLY instead of one antibody
After administration of 5 mg of the synthetic peptide of PELA, T
Administration of SST-1 was started.

In (3), immediately after the administration of the toxin, blood purification was carried out by a conventional method using an extracorporeal circulation method using a column filled with the TSST-1 adsorption fiber prepared in Example 1. Extracorporeal circulation was performed by inserting a double-lumen catheter into the jugular vein to secure blood vessels, and using a circulating pump for dialysis. The flow rate was 80 ml / min, and heparin was used as an anticoagulant.

[0030]

[Table 2]

As described above, administration of the antibody or peptide that detoxifies TSST-1 before the administration of the toxin could prevent a decrease in cardiac function, in particular, the workload of the left ventricle. Also, T
Even after SST-1 administration, it was shown that the column which removes TSST-1 restores reduced heart function, especially the left ventricular workload.

[0032]

Industrial Applicability According to the present invention, a material capable of preventing a decrease in cardiac function or recovering a decreased function by detoxifying or removing a substance that inhibits cardiac function in a solution having a high protein concentration can be provided. By using the material of the present invention, it is possible to detoxify or remove a cardiac function-suppressing substance present in blood, and to provide a drug or a blood purification column capable of treating or preventing a decrease in cardiac function due to sepsis or the like. It became possible to do.

 ──────────────────────────────────────────────────続 き Continued on the front page F term (reference) 4C077 AA01 BB03 EE01 KK11 MM02 MM08 NN14 PP07 PP21 4C084 AA16 BA42 BA44 ZA361 ZA362 ZC801 ZC802 4D017 AA11 BA20 CA11 DA03

Claims (8)

[Claims] 1. A material for treating heart failure, which removes or detoxifies a substance inhibiting cardiac function in blood. 2. The material for treating heart failure according to claim 1, wherein the substance inhibiting cardiac function is a substance derived from bacteria. 3. The material for treating heart failure according to claim 2, wherein the bacterium is Staphylococcus aureus or Group A streptococcus. 4. A material for treating heart failure wherein the cardiac function-suppressing substance is a superantigen. 5. The material for treating heart failure according to claim 1, wherein the cardiac function-suppressing substance is a left ventricular function-suppressing substance. 6. The material for treating heart failure according to claim 1, which has a urea bond, a thiourea bond or an amide bond. 7. The material for treating heart failure according to claim 1, which is water-insoluble. 8. A blood purification column filled with the material for treating heart failure according to any one of claims 1 to 7.