JPS6367858B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a blood coagulation promoter that has the effect of promoting coagulation of collected blood, and more specifically, for the purpose of separating serum in a short time or determining the clotting time in a blood coagulation test in a short time. The present invention relates to a blood coagulation promoter that is used by mixing it into collected blood. Coupled with the remarkable progress in preventive medicine in recent years, blood tests such as serum biochemistry tests, serum immunology tests, and hematology tests have become widespread. Serum tests in particular form the main body of blood tests, and the serum required for testing is After the blood collected in a blood test container is coagulated, it is centrifuged to separate and collect blood clots (gel-like lumps formed by mixing fibrin and blood cells) with different specific gravities. Blood coagulation tests determine various bleeding disorders, such as vascular dysfunction diseases, platelet dysfunction diseases, and coagulation disorder diseases, by evaluating the activity of various factors related to blood coagulation or platelet function. It has practical significance as an important clinical testing method. Various conventional blood coagulation tests are based on measuring the coagulation time of blood collected from a specimen. In the above serum test and blood coagulation test,
The problem that has been considered in the past is that the test is carried out within a short period of time after blood is collected from the specimen, making it impossible to obtain measured values.This is because it takes a considerable amount of time for the collected blood to coagulate. It was necessary to do so. To solve the above problems, conventionally, kaolin, celite, etc. were added to the blood collected in blood test containers.
A method of adding inorganic powder such as glass, silicon oxide, or bentonite to accelerate coagulation, a method of adding small spheres or pellets of organic synthetic resin such as polystyrene, or a method of adding a substance with coagulation-promoting activity extracted from living organisms, such as snake venom. For example, methods using thrombin-like substances obtained from animal brains and thromboplastin substances extracted from animal brains have been used. However, the methods using inorganic powders or organic synthetic resin globules or pellets have problems such as insufficient blood coagulation time shortening effects and hemolysis when used in large quantities. In addition, the above-mentioned methods using extracts from living organisms have problems such as the fact that they tend to lose their activity during storage and are expensive. In order to solve the above-mentioned problems, the present inventors have earnestly searched for substances that are effective for activating coagulation factors and platelets in blood coagulation. As a result, it was discovered that certain catechol derivatives have superior activity to factors such as kaolin, celite, glass, and ellagic acid, which have been known to have effective effects on factor XII and platelets in blood coagulation. It came to this. The gist of the present invention is as follows: 1 formula [In the formula, for R ₁ , R ₂ , and R ₃ , the following (a), (b),
Either (c). (a) One or two of R ₁ , R ₂ , and R ₃ is an alkyl group having 1 to 8 carbon atoms, and the others are hydrogen atoms; (b) R ₁ is an alkyl group having 1 to 8 carbon atoms;

[Formula] is a group in which R ₂ and R ₃ are hydrogen atoms, and (c) R ₁ is

A blood coagulation promoter, characterized in that it consists of a catechol derivative represented by the following formula: 2. 1 part by _weight of the catechol derivative and glass, silica, kaolin _, Adsorptive inorganic substance selected from the group consisting of celite and bentonite 0.05 to
A blood coagulation promoter, characterized in that it consists of 10 parts by weight. Next, the blood coagulation promoter of the present invention will be explained in more detail. The blood coagulation promoter of the present invention consists of a catechol derivative represented by the following formula. In the case of the above formula, the catechol derivative is R ₁ ,
They are classified as follows depending on the types of R ₂ and R ₃ . (1) 3-Alkylcatechol This is the case when R ₁ is an alkyl group having 1 to 8 carbon atoms and R ₂ and R ₃ are hydrogen atoms, such as 3-propylcatechol, 3-normal Butylcatechol, 3-tertiarybutylcatechol, 3-n-hexylcatechol, 3-
2-ethylhexylcatechol and the like exist. (2) 4-Alkylcatechol This applies when R ₂ is an alkyl group having 1 to 8 carbon atoms, and R ₁ and R ₃ are hydrogen atoms, such as 4-propylcatechol, 4-normal Butylcatechol, 4-tertiarybutylcatechol, 4-n-hexylcatechol, 4-
2-ethylhexylcatechol and the like exist. (3) Para-alkylcatechol This is the case when R ₂ and R ₃ are alkyl groups having 1 to 8 carbon atoms and R ₁ is a hydrogen atom, for example, 1-ethyl-4-tert-butyl Catechol, 1.4-ditertyabutylcatechol,
There are 1.4-dihexylcatechol, 1.4-2-ethylhexylcatechol, etc. (4) Catechin

[Formula] This is a catechol derivative represented by the formula on the left. Catechins are widely distributed in tree wood, twigs, bark, leaves, etc., and are particularly abundant in gambia and betel nut. It is colorless fine needle-shaped crystals, and those recrystallized from water contain 4 molecules of water of crystallization. Decomposition points are 93-95
℃, the melting point of anhydride is 96℃, and the decomposition point is 174~
The temperature is 175℃. The absorption maximum is 280 mμ. (5) Nordihydrogayaretic acid It is a catechol derivative represented by the above formula. Nordihydrogayaretic acid is a substance obtained as the main component of resin from the leaves and stems of Tribulaceae plants, or is a substance obtained by synthesis, and is a white or pale yellow crystal with a melting point of 184-185°C. Slightly soluble in water, insoluble in petroleum ether, easily soluble in ethanol, ether, and acetic acid. Among the above catechol derivatives, those which are particularly excellent as blood coagulation promoters include 3-tertiarybutylcatechol, 4-tertiarybutylcatechol, paratertiarybutylcatechol, catechin, nordihydrogayaretic acid, etc. be. As the blood coagulation promoter in the present invention, the above-mentioned catechol derivative can be added to blood as it is. It can also be used in combination with any other substance that does not inhibit blood coagulation promotion or interfere with blood tests. Substances that can be used in combination with the blood coagulation promoter include polymeric substances, oligomeric substances, and low-molecular substances that do not have the effect of denaturing blood. Examples of polymeric substances and oligomeric substances include polystyrene, polyethylene, polypropylene, polyvinyl chloride, polymethyl methacrylate, polymethyl acrylate, polyethyl acrylate, polyacrylonitrile, polybutadiene, polybutene, polydimethylsiloxane, paraffin, chlorinated paraffin, and chlorine. Used are chlorinated polyethylene, chlorinated polybutene, polyethylene glycol, cellulose, cellulose acetate, chitin, and the like. Examples of low-molecular substances that do not have the effect of denaturing blood used in the present invention include higher aliphatic alcohols such as lauryl alcohol, palmityl alcohol, and stearyl alcohol;
Polyhydric alcohols such as glycerin and sorbitan; esterified products of polyhydric alcohols such as glycerin monostearate, glycerin distearate, and sorbitan monostearate; vegetable and animal oils and fats are preferably used. The blood coagulation promoter of the present invention may be in either solid or liquid form. If it is in solid form, it can take any form such as powder, spherules, pellets, etc. When taking the shape of a small sphere or pellet, the blood coagulation promoter of the present invention may be attached to the surface of a carrier that has been previously formed into the shape of a small sphere or pellet using the above-mentioned polymeric substance, or alternatively,
After the blood coagulation promoter of the present invention is uniformly mixed with the above-mentioned polymeric substance, it can be formed into the shape of small spheres or pellets. In the latter case,
In order for the blood coagulation promoter of the present invention present in the base polymeric substance to migrate to the surface of the molded globules or pellets and to effectively promote blood coagulation, the necessary amount must be present on the surface. It is preferable to use compounds that promote surface migration, such as higher aliphatic alcohols, polyhydric alcohols, fatty acid esters, higher aliphatic carboxylic acids, and hydrocarbon waxes. When the blood coagulation promoter of the present invention is in a solid state, its particle size can be set arbitrarily, but it is preferable that it has a large contact area with blood.
A diameter in the range of 1 micron to 10 mm is preferred. When the blood coagulation promoter of the present invention is in liquid form, water, methyl alcohol, ethyl alcohol,
It is preferable to dissolve or disperse in ethylene glycol, glycerin, etc., and high viscosity liquids, such as polymer oligomers such as polyethyl acrylate oligomer, liquid polyptadiene, liquid paraffin, chlorinated paraffin, chlorinated polybutene, polyethylene glycol, silicone oil, etc. It can be used by mixing it with oil. In the present invention, the effective amount of the catechol derivative added to blood can be selected as desired depending on the desired coagulation rate, but a particularly preferred range is 0.01 mg to 10 mg per 5 ml of blood volume. In the present invention, when the catechol derivative is used alone, a significant blood coagulation promoting effect is observed. However, it has been found that when the catechol derivative and the adsorbent inorganic substance are used together, the blood coagulation promoting effect is more excellent than when the catechol derivative or the adsorbent inorganic substance is used alone. The adsorbent inorganic substance is selected from the group consisting of glass, silica, kaolin, celite, and bentonite, and these have excellent blood adsorption properties. When the adsorptive inorganic material is in powder form, it is preferable that the weight ratio of the adsorbent inorganic material used is 0.05 to 10 to 1 of the catechol derivative. When the adsorptive inorganic substance is in the form of small spheres or pellets, it is particularly preferable to use one coated with the catechol derivative heated and melted or dissolved in a solvent on the surface thereof. Among the adsorptive inorganic materials, silicate glasses such as soda lime glass, phosphosilicate glass, and borosilicate glass, and quartz glass are used as glasses. In addition, silica is produced by hydrolyzing silicon tetrachloride in an oxyhydrogen flame and has an average particle size of 5 to 5.
Fine silicon dioxide particles of 40 millimicrons, fine silicic acid with an average particle size of 2 to 5 microns produced from sodium silicate, and fine silicic acid with an average particle size of 2 to 10 microns obtained from natural amorphous silicic acid. etc. are suitable. When using the blood coagulation accelerator of the present invention, the blood may be collected into a blood test container such as a spittoon or the like after it has been placed in advance, or it may be used in addition to the blood collected in the blood test container. There is no problem. According to the blood coagulation promoter of the present invention, blood coagulation factors are activated rapidly, the time required for blood coagulation is significantly shortened, and serum and blood clots can be easily separated, separated, and collected. The problem of residual fibrin and blood clot components being mixed into the blood serum has also been resolved.
Further, since the contraction of blood clot components progresses sufficiently, there are advantages such as a significantly large yield of serum. Examples of the present invention are described below. Example 1 3- on the surface of polystyrene beads with a diameter of 2.5 mm
A product with methylcatechol attached was created as a blood coagulation promoter. The attachment of 3-methylcatechol to the surface of the beads was carried out by adding a methanol solution of 3-methylcatechol to the beads, stirring thoroughly, and then drying the methanol. The amount of 3-methylcatechol attached to the bead surface was 0.15 mg per gram of beads. 1 g of the above blood coagulation promoter with an outer diameter of 15 mm and a height of
A 100 mm polymethyl methacrylate spittoon was prepared. Immediately after blood collection, 5 c.c. of blood from a normal healthy person was injected into each spittoon and left at 20°C. After blood collection, the time required for the whole blood to stop flowing completely was measured as the blood coagulation time. Immediately after blood coagulation, perform centrifugation at a rotation speed of 3000 rpm for 5 minutes, observe the state of serum separation, and collect the supernatant serum with a pipette.
The amount was defined as the serum yield. As is clear from the results in the Example 1 column of Table 1, when 3-methylcatechol was used, a remarkable blood coagulation promoting effect was achieved, and the serum separation condition was also very good. Examples 2 to 5 In Example 1, instead of using 0.15 mg of 3-methylcatechol per 1 g of polystyrene beads, 0.23 mg of 4-tertiarybutylcatechol was used per 1 g of polystyrene beads ( Example 2), 0.27 mg of paratertiary liptylcatechol per 1 g of polystyrene beads.
(Example 3), 0.23 mg of catechin per 1 g of polystyrene beads (Example 4), and 0.2 mg of nordihydrogayaretic acid per 1 g of polystyrene beads (Example 4). Example 5) were prepared respectively. Next, the blood coagulation time was measured in the same manner as in Example 1, the state of serum separation was observed, and the serum yield was measured. As is clear from the results of Examples 2 to 5 in Table 1, a remarkable blood coagulation promoting effect was obtained in all cases, and the serum separation condition was also extremely good. Examples 6 to 7 A blood coagulation promoter in the form of a dispersion (Example 6) consisting of 0.06 g of 4-tertiarybutylcatechol, 0.24 g of fine silica powder (average particle size 4 μ), and 100 g of water, and 0.06 g of catechin, and fine silica powder. Powder (average particle size 4μ)
A blood coagulation promoter (Example 7) in the form of a dispersion liquid consisting of 0.24 g and 100 g of water was prepared. When testing these blood coagulation promoters, the dispersion was sufficiently stirred to sufficiently uniformly suspend the precipitated components, and then 0.1 cc was added to each spittoon made of polymethyl methacrylate.
were added one by one. Next, after collecting blood from a normal healthy person,
Immediately injected 5 c.c. and left at 20°C. Next, the blood coagulation time was measured in the same manner as in Example 1, and the state of blood separation was observed. In all cases, a remarkable synergistic effect in promoting blood coagulation was observed, and the blood separation condition was also extremely good. Comparative example 1 1 g of polystyrene beads with a diameter of 2.5 mm and an outer diameter of 15
A container made of polymethyl methacrylate with a height of 100 mm and a height of 100 mm was prepared. Next, the blood coagulation time was measured in the same manner as in Example 1, and the state of serum separation was observed. Table 1 shows the results.
It is described in the column of Comparative Example 1. Comparative Example 2 1 g of small disc-shaped polystyrene beads with a diameter of 3.3 mm and a thickness of 1.8 mm were placed in a polymethyl methacrylate spittoon with an outer diameter of 15 mm and a height of 100 mm. Next, the blood coagulation time was measured in the same manner as in Example 1, and the state of serum separation was observed. Table 1 shows the results.
It is described in the column of Comparative Example 2. Comparative example 3 Colloidal silica (average particle size 5 mμ to 40 m
A blood coagulation promoter was prepared in the form of a dispersion consisting of 1 g of μ) and 100 g of water. A polymethyl methacrylate spittoon containing 0.1 cc of the above blood coagulation promoter was prepared. Next, the blood coagulation time was measured in the same manner as in Example 1, the state of serum separation was observed, and the serum yield was measured. The results are shown in the Comparative Example 3 column of Table 1.

【table】

【table】

Claims (1)

[Claims] 1 formula [In the formula, for R ₁ , R ₂ , and R ₃ , the following (a), (b),
Either (c). (a) One or two of R ₁ , R ₂ , and R ₃ is an alkyl group having 1 to 8 carbon atoms, and the others are hydrogen atoms; (b) R ₁ is a group of [formula]; (c) R ₁ is a group of [Formula], R ₂ and R _{3 are} hydrogen atoms, and R 2 and R ₃ are hydrogen atoms; Accelerator. 2 formulas [In the formula, for R ₁ , R ₂ , and R ₃ , the following (a), (b),
Either (c). (a) One or two of R ₁ , R ₂ , and R ₃ is an alkyl group having 1 to 8 carbon atoms, and the others are hydrogen atoms; (b) R ₁ is a group of [formula]; ₁ part by weight of _{a catechol derivative represented by R 2 and R 3 are hydrogen atoms} ; A blood coagulation promoter characterized by comprising 0.05 to 10 parts by weight of an adsorbent inorganic substance selected from the group consisting of celite and bentonite.