JPS6253665A - Antithrombotic agent - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] [Industrial application field] The present invention relates to anticoagulants.

In recent years, with the development of clinical medicine, blood tests have played an extremely important role in determining the status of preventive medicine or treatment. Under these circumstances, the development of anticoagulants that prevent coagulation of blood collected for blood tests is actively underway.

Further, anti-blood coagulants are being developed to prevent blood from coagulating on the surfaces of materials that come into contact with blood during treatments such as extracorporeal circulation circuits and artificial organs.

The present invention relates to anticoagulants particularly suitable for these purposes.

[Conventional technology]

Heparin sodium is the most well-known anticoagulant, and is used to add blood during artificial dialysis, artificial lung therapy, etc., and to treat surfaces that come into contact with blood.

In addition, metal salts of ethylenediaminetetraacetic acid are used as anticoagulants for certain blood morphology tests.

[Problems to be solved by the invention]

However, since heparin can only be obtained by extraction from animal organs, it is not sufficient in quantity compared to synthetic products, and the manufacturing cost is significantly higher.

In addition, it is difficult to obtain heparin with the same structure and performance when the extraction source organs are different, and it also has the disadvantage that it is difficult to supply at a stable price because it depends on imports.

In addition, although metal salts of ethylenediaminetetraacetic acid can be used as anticoagulants for blood morphology testing,
It has drawbacks such as not being able to quantify inorganic ions, which is a type of biochemical test, and interfering with enzyme tests.
For this reason, blood tests require complicated methods such as selecting and using various anticoagulants depending on each test item and using a serum separation method in which serum is separated from blood before measurement. I was forced to find a way.

Therefore, there is a demand for the development of an anti-coagulant that is inexpensive, has anti-coagulant properties, and does not have an adverse effect on various blood tests, but an anti-coagulant that satisfies these demands has not yet been developed. It has not been.

As a result of extensive research in view of the current situation, the present invention, which relates to an anti-blood coagulant that can be used in various blood tests, has been arrived at.

[Means for solving problems]

That is, the gist of the present invention is that i mol % or more, preferably 5 mol % or more of the primary amino groups in polyallylamine
It is an anticoagulant whose main ingredient is sulfonated polyallylamine, in which 1% or more of the sulfonated polyallylamine is sulfonated.

The sulfonated polyallylamines may be partially or completely neutralized with alkali metal or alkaline earth metal hydroxides.

Polyallylamine is obtained by polymerizing allylamine, and is usually obtained by radical μ polymerization of allylamine inorganic acid salt in an aqueous solution to obtain polyallylamine inorganic acid salt, and then removing the inorganic acid. Hydrochloric acid is usually used. Methods for removing inorganic acids include a method using a basic ion exchange resin, a dialysis method, or a neutralization reaction with a strong base such as caustic soda. There is a method to separate P by precipitating it in it.

Also, 1! of the primary amino group of polyallylamine! n01-
As mentioned above, it is necessary that sulfonation is preferably at least 5 m01. If the degree of sulfonation is less than 1 mol 4, there are too few sulfonated functional groups and it is difficult to fully exhibit anti-blood coagulation properties.

Although this sulfonated polyallylamine bar can be used as it is, that is, in its sulfonic acid form, some or all of the sulfonic acid groups can be neutralized with an alkali metal or alkaline earth metal hydroxide or ammonium hydroxide. It is preferable that it is Japanese.

The molecular weight (weight average) of polyallylamine is 1000-1
It is preferable that it is about 0,000.

The method for nulfonation of polyallylamine to obtain the anticoagulant of the present invention will be described below.

Methods for sulfonation of polyallylamine include a method of reacting with chlorosulfonic acid and sulfonation, and a method of sulfonation with hot 113 sulfuric acid.

The nurefonation method using chlorosulfonic acid can be carried out by dissolving polyallylamine in a solvent such as methano-μ, adding an appropriate amount of chlorosulfonic acid, and causing the reaction. As the solvent, alcohols such as methanol and isopropano-μ, ketones such as acetone and methyμethylketone, and halogenated hydrocarbons such as chloroform, hydrogen tetrachloride, and dichloroethane can be used. The concentration of polyallylamine in the solvent should be 0.5% by weight or more and 50% by weight or less.

If the weight is less than α5 weight, the amount of solvent used becomes too large, making it difficult to recover the polymer after chlorosulfonation. Also, 50
If the concentration exceeds % by weight, it becomes difficult to control the reaction heat during the cross-μ sulfonation reaction. In addition, the amount of chlorosulfonic acid used is 1 (1,0 parts by weight) per 7100 parts by weight of polyamide
It is preferable that the amount is at least 1 part by weight; if it is less than 10.0 parts by weight, the sulfonation reaction will not proceed sufficiently.

However, it is difficult to sulfonate both of the two hydrogen atoms in the primary amino nitrogen of the amino group of polyallylamine, and usually only one of them is sulfonated.

Next, in the sulfonation method using hot concentrated sulfuric acid, 96 to 10
Sulfonation can be achieved by directly adding concentrated sulfuric acid with a purity of 0% by weight to polyallylamine and heating. Here, the degree of disulfonation is determined by the amount of sulfuric acid reacted with polyallylamine and the heating temperature. That is, the amount of sulfuric acid used should be at least 50 parts by weight per 100 parts of polyallylamine.

If the amount is less than 50 parts, sulfonation with sulfuric acid will not proceed effectively. Concentrated sulfuric acid is gradually added to polyallylamine,
It should not be added locally in large amounts to cause a dehydration reaction from polyallylamine. Next, the reaction temperature after adding concentrated sulfuric acid is 100°C or higher and lower than 200°C for 30 minutes to 12 minutes.
Should be added for 0 minutes. At high temperatures, sulfonation is completed in a relatively short time, and at low temperatures, sulfonation is completed in about 120 minutes. The degree of sulfonation can be confirmed by analysis using Fourier transform infrared absorption spectrum μ.

Since the polyallylamine sulfonated compound reacted as described above contains unreacted chlorsulfonic acid, sulfuric acid, etc., these are removed by purification. For example, cine deposits can be removed by dissolving a sulfonated polyallylamine compound in water, dropping it into an alcohol such as methanol-μ, isozolopanol, or a ketone such as acetone or meth-ethyl ketone, reprecipitating it, separating it, and then drying it. I can do it. It is preferable that the amount of water dissolved during this purification be as small as possible. Therefore, in the case of reprecipitation, the concentration of the aqueous solution of the polyallylamine sulfonated compound dropped into alcohol or ketone is 10% by weight or more and 80% by weight or less, preferably 30% by weight or more.
It is preferable that the amount is from - to 60% by weight. 10
When the weight is i%, the amount of the sulfonated compound of polyallylamine that precipitates when added dropwise to Alco-p or ketone is small, resulting in a decrease in recovery rate. Moreover, if the weight exceeds 80%, it becomes difficult to prepare an aqueous solution of the sulfonated compound.

Neutralization of the sulfonated compound of polyallylamine can be done by preparing an aqueous solution of the sulfonated compound and neutralizing it by adding an aqueous solution of the specified alkali metal or alkaline earth metal hydroxide or ammonium hydroxide. . Examples of the neutralizing agent used here include sodium hydroxide, potassium hydroxide, magnesium hydroxide, barium hydroxide, ammonium hydroxide, and the like. A reaction product obtained by neutralizing a sulfonated compound with a neutralizing agent can also be purified by a reprecipitation method using alcohol-μ or the like.

The anticoagulant of the present invention can use a sulfonated compound of polyallylamine, a portion thereof, or a completely neutralized product thereof, either alone or in combination. At this time, it may contain trace amounts of heparin salt, oxalate, oxalate complex salt, citrate, etc., which do not affect blood test results.

〔Example〕

The present invention will be explained in more detail with reference to Examples below.

In the Examples, the degree of sulfonation of the sulfonated polyamine was determined by Fourier transform infrared absorption spectrum analysis.

Example 1 Polyallylamine with molecular weight IQ of Goo (PAA-HCL-L manufactured by Nittobo Co., Ltd.) 5 I! Methanol/L15
It was dissolved in 0-. To this was gradually added 20 μm of Croxuphonic acid. After finishing the addition, while stirring 6.
The reaction was completed by raising the temperature to 0° C. and maintaining the reaction rate for 30 minutes. At this time, the sulfonation reaction product of polyallylamine precipitated as a substance insoluble in the solvent, so it was separated by nine-mouth filtration using a glass filter. 5 cc of water was added to this product and the aqueous solution was stirred.
/I/200- by gradually dropping it into the solution. The generated reaction product was separated and dried under reduced pressure to obtain crystalline powder 58y. When the degree of sulfonation of this powder was measured, 38 of the -NH groups of polyallylamine were sulfonated. A portion of this powder was collected and dissolved in water to prepare a 10 wt % aqueous solution.

5 μL, 2 μL, and 1 μL of this 10 wt aqueous solution were collected and placed in test tubes, respectively, and approximately 1 μL of fresh blood was added thereto.

When the presence or absence of blood clots was visually measured 3 hours after addition, no blood clots occurred in any of the samples /L/.

When the blood of these samples was observed using an optical microscope, no changes were observed in red blood cells, white blood cells, platelets, etc.

Example 2 Next, various amounts of crosμ-μphonic acid were investigated. At this time, the amount of polyallylamine as a raw material was 5.0 g, and the reaction temperature, reaction time, purification method, etc. were the same as in Example 1. The results are shown in Table 1.

From Table 1, it was found that when the amount of chlorosulfonic acid added was reduced and the degree of sulfonation was less than 1% (comparative example), it did not act as an anticoagulant.

Table 1 Judgment of pot anticoagulability was performed as follows.

(1) 1vt of sulfonated polyethyleneimine
Put 100μt of % aqueous solution into a test tube and add fresh blood to it.
After adding gIt and mixing several times, let it stand. (2) Visually observe the presence or absence of blood clots at 30 minute intervals for 5 hours.

When blood did not coagulate, it was given an X when some Ol aggregation was observed.

(3) For blood that did not coagulate, the presence or absence of deformation in the morphology of platelets and blood cells was examined using an optical microscope.

Example 3 Sulfonated product 1 of polyallylamine prepared in Example 1
g was collected and dissolved in 9 cc of water to create a 10 ml aqueous solution of the reaction product.

To this was added 2.2 td of 4N aqueous sodium hydroxide solution for neutralization. This aqueous solution was gradually dropped into 400 d methanol-μ for recrystallization. The generated reaction product was separated and dried under reduced pressure to obtain a crystalline powder CL65J.

This crystalline powder was subjected to the same anticoagulant test as in Example 2. 1. of crystalline powder. Owt%
50 μL of the aqueous solution was collected in a test tube and approximately 1.0 μL of blood was added to measure blood coagulation, but no blood coagulation was observed even after 5 hours.

Further, when this sample μ was observed under a microscope, no aggregation of red blood cells, white blood cells, or platelets was observed.

Example 4 Polyaryl with OOO molecular weight of 1! Amine 5F was collected, and 5 g of concentrated sulfuric acid with a purity of 98 wt was gradually added dropwise thereto and mixed. The sulfuric acid solution was then heated to 120°C and maintained at this temperature for 90 minutes. At this time, stirring of the sulfuric acid solution was maintained to prevent local heating. Add 10 c of water to this reaction solution.
After adding and dissolving c, the reactant was recrystallized by dropping it into methanol-A1500, and the remaining sulfuric acid was removed by methanol-μ. The separated reaction product was dried under reduced pressure to obtain 2.7 g of crystalline powder.

The degree of sulfonation of this reaction product was measured and found to be 22. A tOvt% aqueous solution of this crystalline powder was prepared, 100μt and 150μt were collected into test tubes, and blood was collected at approximately 1% each. The blood coagulability was measured by adding low. No clots were observed even after 5 hours of blood addition, and these samples ~
Microscopic observation of the patient revealed no abnormalities in red blood cells, white blood cells, or platelets.

〔Effect of the invention〕

The anti-blood coagulant of the present invention can be easily synthesized using polyallylamine, a chemical product, is cheaper and more abundant than conventional anti-blood coagulants, and exhibits sufficient anti-blood coagulability. It has the characteristic of not causing shape deformation.

Claims (1)

[Scope of Claims] 1. An anti-blood coagulant whose main component is a sulfonated polyallylamine in which 1 mol% or more of -NH_2 groups in the polyallylamine are sulfonated. 2. The anti-blood coagulation according to claim 1, wherein the sulfonated polyallylamine is at least partially neutralized with an alkali metal or alkaline earth metal hydroxide or ammonium salt. agent.