JPH0152027B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing an adsorbent that has the ability to adsorb albumin-binding substances that are present in large amounts in the blood of patients suffering from liver failure or various drug addictions.

In cases of liver failure, etc., the amount of metabolically abnormal substances increases in the patient's blood, and as the disease progresses, so-called hepatic coma is induced. As a treatment for such liver failure, auxiliary liver therapy has been attempted to detoxify patients through extracorporeal blood perfusion using adsorbents such as activated charcoal, and has been shown to be effective in helping patients wake up from coma. . However, activated carbon has a low adsorption capacity for albumin-binding substances, and it remains difficult to increase the survival rate. In addition, adsorbents in which albumin is immobilized on particles such as agarose are known for removing albumin-binding substances such as bilirubin, but these particles have a small amount of albumin immobilized, and the albumin-binding substances The adsorption capacity of is not sufficient. Furthermore, attempts have been made to use anion exchange resins or various synthetic adsorbents, but these all have insufficient adsorption capacity for albumin-binding substances.
It has not yet been put into practical use. Therefore, it is desired to develop a new adsorbent with high adsorption efficiency for albumin-binding substances.

The present inventor conducted extensive research to develop an adsorbent that can efficiently adsorb and remove harmful substances in the blood, such as bilirubin, which has albumin binding properties, and found that a polyvalent amine compound and a polyvalent epoxy The present invention was achieved by discovering that a polymer prepared by suspension polymerizing a compound in an aqueous medium under specific conditions can efficiently adsorb and remove albumin-binding substances in blood.

That is, in the present invention, a polyvalent amine compound and a polyvalent epoxy compound are subjected to a polyaddition reaction in an inert organic solvent immiscible with water at a temperature of less than 30°C, and then, if necessary, the polyvalent epoxy compound is By adding, all the epoxy groups in the polyvalent epoxy compound used in the reaction are reduced to one of the amine groups and imino groups in the polyvalent amine compound used in the reaction.
1.05 to 1.70 per equivalent or per equivalent of amine group if the polyvalent amine compound does not contain an imino group
An adsorbent that consists of crosslinking and hardening the suspended particles by adjusting the weight to an equivalent range, suspending and dispersing them in an aqueous medium containing a dispersion stabilizer, and further reacting at a temperature of 30°C or higher. This is a manufacturing method.

The present invention will be explained.

Non-limiting examples of the polyvalent amine compound which is a necessary component for producing the adsorbent of the present invention include the following compounds.

H ₂ N−(CH ₂ −) _o NH ₂ .R—NH−(CH ₂ CH ₂ NH
-) _n R'. [However, n represents an integer of 1 to 10, m represents an integer of 1 to 10,000, and R and R' independently represent a hydrogen atom or alkyl, alkenyl, aryl, having 1 to 20 carbon atoms, Represents an aralkyl or alkanol. ] etc.

Non-limiting examples of polyepoxy compounds used in the production of the adsorbent of the present invention include the following materials:

or (However, p, q, r represent integers from 1 to 10.)
or glycerol diglycidyl ether, glycerol triglycidyl ether, 1,1,1-trimethylolpropane triglycidyl ether,
Neopentyl glycol diglycidyl ether,
Sorbitol polyglycidyl ether, hydroquinone diglycidyl ether, resorcin diglycidyl ether, triglycidyl isocyanurate, etc.

In the method for producing the adsorbent of the present invention, first, a polyvalent amine compound and a polyvalent epoxy compound are subjected to a polyaddition reaction in an inert organic solvent immiscible with water at a temperature of less than 30°C. After preparing the linear prepolymer, the total amount of epoxy groups used in the reaction is adjusted to be in excess of the amine groups and/or imino groups in the reaction mixture, and then this is mixed with a dispersion stabilizer. Suspension and dispersion in an aqueous medium of 30
Spherical particles are produced by crosslinking and curing by heating to a temperature of .degree. C. or higher.

In this case, in the first stage prepolymerization in which the reaction is carried out at a temperature below 30°C, the quantitative ratio of amine groups and/or imino groups to epoxy groups can be either excessive epoxy groups or excessive amine groups. There isn't. Of course, it is preferable that the degree of polymerization of the prepolymer is higher, since this increases the mechanical strength of the adsorbent, and therefore it is preferable that the molar ratio of the amine groups to the epoxy groups is approximately equimolar. Regarding the amount of reaction reagents to be charged, the reaction can be carried out by first adding the entire amount of excess polyvalent epoxy compound and polyvalent amine compound into a reaction vessel. In addition, by first adding equimolar amounts of a polyvalent epoxy compound and a polyvalent amine compound and causing the reaction, the degree of polymerization is sufficiently increased, and then an excess amount of the polyvalent epoxy compound is added to the reaction mixture and used for the reaction. It is also possible to adjust the total amount of epoxy groups to be 1.05 to 1.70 equivalents per equivalent of amine and/or imino groups in the reaction mixture.

When the first step of prepolymerization for prepolymer preparation is completed in this way, the prepolymer solution is suspended and dispersed in an aqueous medium containing a dispersion stabilizer, etc., and the reaction temperature is increased to 30°C or higher. Spherical adsorbent particles are obtained by raising and crosslinking and hardening the suspended particles.

The reaction temperature at which the polyaddition reaction is carried out in the production of the adsorbent of the present invention is such that the reaction is carried out at a temperature of less than 30°C during the preliminary polymerization for preparing the prepolymer in the first stage. At temperatures below 30°C, epoxy groups and amine groups (primary amine groups) react preferentially, and the reaction between epoxy groups and imino groups (secondary amine groups) is suppressed, so linear prepolymers is selectively produced, and the production of branched prepolymers that lead to crosslinking and curing reactions can be suppressed.

Next, in the second step, the crosslinking and curing reaction is carried out by suspending and dispersing the prepolymer solution in an aqueous medium, and then raising the temperature to 30°C or higher. A reaction can occur to crosslink and harden the suspended particles. This crosslinking curing reaction is carried out at a temperature range of 30°C to 100°C, preferably 40°C to 80°C, particularly preferably 45°C to 70°C.
The reaction can be carried out at a temperature of °C.

In addition, the total epoxy group used in the reaction is per equivalent of amine group and/or imino group in the reaction mixture.
It ranges from 1.05 to 1.70 equivalents. If the total amount of epoxy groups is less than 1.05 equivalents per equivalent of amino groups and/or imino groups, the amount of crosslinking will be insufficient and the curing of the adsorbent particles will be insufficient, making it impossible to obtain spherical particles with high strength. On the other hand, if the total amount of epoxy groups exceeds 1.70 equivalents per equivalent of amine group and/or imino group, the amount of crosslinking will be too large and will prevent the adsorbed substance from diffusing into the adsorbent particles, reducing the adsorption efficiency. This is not desirable because it becomes impractical.

The water-immiscible inert organic solvent used in the production method of the present invention is optional, but it is necessary that it has the ability to dissolve the polyaddition reaction product between the polyvalent amine compound and the polyvalent epoxy compound. be. Examples of such solvents include halogenated aliphatic hydrocarbons such as methylene chloride, chloroform, dichloroethanes, trichloroethanes, trichloroethylenes, and tetrachloroethanes; halogenated aromatic hydrocarbons such as monochlorobenzene and dichlorobenzene; Non-limiting examples include aliphatic hydrocarbons such as hexane, heptane, octane, and cyclohexane, aromatic hydrocarbons such as benzene, toluene, and xylenes, and mixtures thereof.

Furthermore, when using a solvent with relatively high specific gravity such as halogenated hydrocarbons, sodium chloride, sodium sulfate, sodium phosphate, etc. are added to the dispersion medium to ensure smooth dispersion with water, which is the dispersion medium. of inorganic salts in an amount of 1% to 50% by weight, preferably 5% by weight.
It is desirable to add and dissolve amounts ranging from % to 20% by weight.

Examples of the dispersion stabilizer used when suspending and dispersing the prepolymer solution obtained by the polyaddition reaction of a polyvalent amine compound and a polyvalent epoxy compound in an aqueous medium in the production method of the present invention include bentonite,
Examples include talc, barium sulfate, calcium carbonate, pectin, gelatin, polyacrylic acid, polyacrylic acid, polyvinyl alcohol, and the like.

The concentration of a mixture of a polyvalent amine compound and a polyvalent epoxy compound or a prepolymer obtained by a polyaddition reaction of these in an organic solvent is 2% by weight to 80% by weight,
Preferably 5% to 60% by weight, particularly preferably
It ranges from 10% to 50% by weight.

The volume ratio of the prepolymer solution obtained by the reaction of the polyvalent amine compound and the polyvalent epoxy compound to the aqueous dispersion medium in which it is suspended and dispersed is 1:2 to 100, preferably 1:2.5 to 50, particularly preferably 1. :3～
The range is 10.

In this way, spherical particles of polyaddition polymer are obtained. The particle size of these spherical particles can be adjusted by adjusting the stirring speed, etc., and is preferably in the range of 0.1 to 2 mm (preferably 0.5 to 1.5 mm) in diameter.

The optimum stirring speed varies depending on the viscosity of the prepolymer solution, the ratio of the prepolymer solution to the dispersion medium, etc., but is generally 50 to 1000 rpm, preferably
A range of 100 to 500 r.pm, particularly preferably 120 to 300 r.pm, is preferably selected.

In the production method of the present invention, the spherical particles of the adsorbent can be made more porous by adding a pore opening agent such as dioctyl phthalate or paraffin to the organic solvent solution of the prepolymer.

Although the adsorbent produced by the method of the present invention can be used in any manner, a method of filling the adsorbent particles into a column or the like and perfusing it with blood, plasma, etc. is preferably used. In particular, one of the major features of the adsorbent produced by the present invention is that blood can be brought into direct contact with the adsorbent and perfused without forming thrombus or the like.

The adsorbent produced according to the present invention can contain albumin-binding substances found in blood that are difficult to remove by operations such as activated carbon adsorption or dialysis, such as thyroxine, triiodothyronine, bilirubin, uric acid, bile acids, guanidine, and indole compounds. It is extremely useful as an adsorbent for acetylcholine, barbituric acid, digitoxin, salicylic acid, and various other drugs.

The present invention will be explained below with reference to Examples. Note that "%" in the examples is based on weight, and the bilirubin concentration in plasma was measured by the Evelyn-Molloy method.

In addition, the concentrations of indocyanine green (ICG) and bromosulfophthalein (BSP) were measured by the absorption intensity of electronic spectra.

Example 1 Bisphenol-A-diglycidyl ether was added to two three-necked flasks equipped with a stirrer and a thermometer.
19.0g (0.05mol), glycerol diglycidyl ether 39.2g (0.2mol), hexamethylene diamine
32.4g (0.28mol) was dissolved in 50ml of chloroform and added, and dioctyl phthalate was added as a pore opening agent.
A prepolymer solution is obtained by adding 20 g and reacting at a reaction temperature of 28° C. for 3 hours. Next, a solution of 10 g of triglycidyl isocyanurate dissolved in 30 ml of chloroform was added to this, and the reaction temperature was 28°C.
After reacting for 1 hour at °C, 40 g of sodium chloride,
Add 800 ml of an aqueous solution containing 6 g of bentonite and suspend and disperse at a stirring speed of 130 rpm.

Next, the reaction temperature was raised to 50°C and the reaction was carried out for 1 hour, and then the reaction temperature was gradually raised to 70°C and the reaction was carried out for 2 hours while distilling off the solvent chloroform. After the reaction was completed, the spherical polymer was separated and further extracted with methanol, washed, and washed with water to obtain 80 g of adsorbent.

After immersing this adsorbent in a phosphate buffer solution with a pH of 7.4, 2 ml was collected, and bilirubin was 10 mg/dl.
After soaking in 20 ml of bovine serum containing 1 mg/dl of indocyanine green (ICG) and 5 mg/dl of bromosulfophthalein (BSP) and shaking at a temperature of 37°C for 3 hours, the adsorbent was separated. When we measured the concentration of each component remaining in bovine serum and determined the adsorption rate, we obtained adsorption rates of 50% for bilirubin, 41% for ICG, and 48% for BSP. It was recognized that there was. Furthermore, this adsorbent
20 ml was packed into a silicone-treated glass column, and extracorporeal blood perfusion was performed for 3 hours at a flow rate of 20 ml/min using an approximately 3.0 kg domestic rabbit that had undergone carotid arteriovenous shunt surgery, and then the blood was returned. As a result of observing the state of the adsorbent, no formation of thrombi or the like was observed, and blood compatibility was good.

Comparative Example 1 Bilirubin, indocyanine green (ICG), In a shaking adsorption test of bromosulfophthalein (BSP), cellulose-coated activated carbon had an adsorption rate of 0%, and Amberlite
XAD-7 is bilirubin 30%, ICG 35%, BSP15
% of each adsorption rate.

Example 2 An adsorbent made of spherical polymer particles was produced in the same manner as in Example 1, except that 34.8 g of ethylene glycol diglycidyl ether was used instead of glycerol diglycidyl ether, and all other conditions were the same.

Collect 2 ml of this adsorbent and use the same method as in Example 1 to detect bilirubin, indocyanine green (ICG), and promosulfophthalein (BSP) in bovine serum.
When a shaking adsorption test was conducted, bilirubin 60%,
Adsorption rates of ICG 60% and BSP 65% were obtained.

Example 3 In the same manner as in Example 1, bisphenol-A
-Diglycidyl ether 19.0g, 1,4-butanediol diglycidyl ether 40.4g, 4,4'-
A prepolymer solution was prepared by reacting 27.6 g of diaminodiphenylmethane and 16.2 g of hexamethylene diamine in the presence of 40 ml of chloroform, 20 ml of cyclohexane, and 20 g of dioctyl phthalate at a temperature of 28°C, and this was mixed with 40 g of sodium chloride and calcium carbonate. 10 g of triglycidyl isocyanurate after suspension dispersion in an aqueous medium containing 10 g of
A solution consisting of 20 ml of chloroform and 10 ml of cyclohexane was added, and a crosslinking and curing reaction was carried out at a temperature of 50 to 70°C to obtain 90 g of an adsorbent made of a spherical polymer.

Collect 2 ml of this adsorbent and use the same method as in Example 1 to detect bilirubin, indocyanine green (ICG), and promosulfophthalein (BSP) in bovine serum.
When a shaking adsorption test was conducted, bilirubin was 75%,
Adsorption rates of ICG 75% and BSP 81% were obtained.

Example 4 In the same manner as in Example 1, bisphenol-A
- Reaction of 19.0 g of diglycidyl ether, 34.8 g of ethylene glycol diglycidyl ether, 27.6 g of 4,4'-diaminodiphenylmethane and 16.2 g of hexamethylene diamine in the presence of 40 ml of chloroform and 20 g of dioctyl phthalate at a temperature of 28°C. A prepolymer solution was prepared, and this was suspended and dispersed in 800 ml of an aqueous medium containing 40 g of sodium chloride and 6.0 g of bentonite by stirring at a stirring speed of 150 rpm, followed by 10 g of triglycidyl isocyanurate and chloroform. Add a solution consisting of 30 ml
A crosslinking and curing reaction was carried out at a temperature of 50 to 70°C to obtain 85 g of an adsorbent made of a spherical polymer.

Using 2 ml of this adsorbent, bilirubin, indocyanine green (ICG), and promosulfophthalein (BSR) in bovine serum were detected in the same manner as in Example 1.
When a shaking adsorption test was conducted, bilirubin was 65%,
Adsorption rates of ICG 62% and BSP 61% were obtained. Furthermore, good blood compatibility was confirmed in an extracorporeal blood perfusion test using rabbits that had undergone carotid artery shunt surgery.

Example 5 In the same manner as in Example 1, 20 g of triglycidyl isocyanurate, 34.8 g of ethylene glycol diglycidyl ether, and hexamethylene diamine were added.
A prepolymer solution was prepared by reacting 31.0 g of stearylamine and 17.2 g of stearylamine in the presence of 40 ml of chloroform and 20 g of paraffin at a temperature of 28° C., which was dissolved in 800 ml of an aqueous medium containing 40 g of sodium chloride and 6.0 g of bentonite. After suspension and dispersion by stirring at a stirring speed of 150 rpm, a solution consisting of 10 g of triglycidyl isocyanurate and 30 ml of chloroform is added, and a crosslinking reaction is carried out at a temperature of 50 to 70°C to form an adsorbent made of spherical polymers. 100g of the drug was obtained.

Using 2 ml of this adsorbent, bilirubin, indocyanine green (ICG), and promosulfophthalein (BSP) in bovine serum were detected in the same manner as in Example 1.
A shaking adsorption test was conducted, and bilirubin 70%, ICG68
% and eyebrow SP70% were obtained. It was found that the protein-binding substance of the present invention was well adsorbed.

Claims (1)

[Claims] 1 Polyvalent amine compound and polyvalent epoxy compound are subjected to a polyaddition reaction in an inert organic solvent immiscible with water at a temperature of less than 30°C, and then the polyvalent epoxy compound is added as necessary. Therefore, all the epoxy groups in the polyvalent epoxy compound used in the reaction are per equivalent of the amine groups and imino groups in the polyvalent amine compound used in the reaction, or the polyvalent amine compound does not contain imino groups. In case, the amount is adjusted to be in the range of 1.05 to 1.70 equivalents per equivalent of amino group, suspended and dispersed in an aqueous medium containing a dispersion stabilizer, and further reacted at a temperature of 30°C or higher. A method for producing an adsorbent comprising crosslinking and curing particles.