JPS6367660B2 - - 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 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, conventional methods include adding inorganic powders such as kaolin, celite, glass, silicon oxide, bentonite, etc. to blood collected in a blood container to promote coagulation, and methods using organic synthetic resins such as polystyrene. Methods that include adding spheres or pellets, or using substances with procoagulant activity extracted from living organisms, such as thrombin-like substances obtained from snake venom 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 problems, we have conducted an intensive search for substances that are effective for activating coagulation factors and platelets in blood coagulation. As a result, 2,2',4,4'-tetrahydroxybenzophenone was found to be more effective than kaolin, celite, glass, ellagic acid, etc., which have been thought to have an effective effect on factor XII and platelets in blood coagulation. It was also discovered that the active ingredient has even better activity. The gist of the present invention is as follows: 1. A blood coagulation promoter characterized by comprising 2,2',4,4'-tetrahydroxybenzophenone.2 2,2',4,4'-tetrahydroxybenzophenone1. 1 part by weight of 2,2',4,4'-tetrahydroxybenzophenone, glass, silica, kaolin,
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. Next, the blood coagulation promoter of the present invention will be explained in more detail. 2,2',4,4'-tetrahydroxybenzophenone (hereinafter abbreviated as 2,2',4,4'-THBP) has the following chemical structural formula. 2,2',4,4'-THBP has a melting point of 195℃, and its solubility in solvents is 0.1% by weight in water, 50% by weight in methanol, 40% by weight in ethanol, and the solubility in water-ethanol at 30℃. 10% by weight for a 1:1 solution. Further, the maximum absorption wavelength position is 345 mμ, and the color value (Gardner) is No. 8 in a 1% by weight methanol solution. And 2, 2', 4,
4′-THBP is a synthetic resin used as an ultraviolet absorber.
Although it was known that it could be incorporated into cosmetics, etc., it was completely unknown that it had applicability as a blood coagulation promoter. The blood coagulation promoter in the present invention is 2,2',4,
4′-THBP can be added to blood alone. 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 used in the present invention that do not have the effect of denaturing blood 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 stearate, 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 the blood coagulation promoter of the present invention is in the form 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 with the above-mentioned polymeric substance. After the inventive blood coagulation promoter is uniformly mixed with the above-mentioned polymeric substance, it can be formed into spherules or pellets. In the latter case, the blood coagulation promoter of the present invention present in the base polymeric substance is
In order for the particles to migrate to the surface of the molded globules or pellets and to effectively promote blood coagulation, it is necessary to present them in the necessary amount on the surface, and compounds that promote surface migration, such as higher aliphatic alcohols and It is particularly preferable to use alcohols, fatty acid esters, higher aliphatic carboxylic acids, hydrocarbons, waxes, etc. in combination. When the blood coagulation promoter of the present invention is in a solid state, its particle size may be arbitrary, 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, for example, polyethyl acrylate oligomer,
It can be used by being mixed into a polymer oligomer such as liquid polybutadiene, liquid paraffin, chlorinated paraffin, chlorinated polybutene, polyethylene glycol, or oil such as glycerin or silicone oil. In the present invention, the effective amount of 2,2',4,4'-THBP 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 2,2',4,4'-THBP is used alone, a significant blood coagulation promoting effect is observed. However, when 2,2′,4,4′-THBP and ellagic acid are used together, and when 2,2′,4,4′-
When using THBP and adsorbent inorganic substances, 2.
It was found that 2',4,4'-THBP has a more excellent blood coagulation promoting effect than when used alone. One of the substances that exhibits a synergistic blood coagulation promoting effect when used in combination with 2,2',4,4'-THBP is ellagic acid. Elazinic acid is a substance with the following chemical structural formula. Ellagic acid is known as a substance that activates factor XII, which is one of the blood coagulation factors.
It was confirmed that when used in combination with 2',4,4'-THBP, it has a more excellent blood coagulation promoting effect than when each of these is used alone. In this case, the weight ratio of ellagic acid used is 2,2′,
4,4'-THBP is set to 0.1 to 5.0 for 1. Other substances that synergistically exhibit a blood coagulation promoting effect when used in combination with 2,2',4,4'-THBP are adsorbent inorganic substances. The adsorbent inorganic substance is selected from the group consisting of glass, silica, kaolin, celite, and bentonite, and these have excellent blood adsorption properties. These substances were also used as blood coagulation promoters, but when used together with 2,2',4,4'-THBP,
It was confirmed that they have a more excellent blood coagulation promoting effect than when each of these is used alone. 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 for 1 of 2,2',4,4'-THBP. In addition, when the adsorptive inorganic substance is in the form of small spheres or pellets, it is particularly recommended to apply 2,2',4,4'-THBP to its surface by melting it by heating or by dissolving it in a solvent. preferable. When using the blood coagulation promoter of the present invention, the blood may be collected into a blood container such as a spittoon in advance after it has been placed in the blood container, or it may be used in addition to the blood collected into the blood container. . 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 collected serum is also solved, and furthermore, the blood clot components are sufficiently contracted, so the yield of serum is significantly increased. Examples of the present invention are described below. Example 1 2, on the surface of polystyrene beads with a diameter of 2.5 mm
A product with 2',4,4'-THBP attached was prepared as a blood coagulation promoter. To attach 2,2',4,4'-THBP to the bead surface, add an ethanol solution of 2,2',4,4'-THBP to the beads, stir thoroughly, and then dry the ethanol. It was done by The amount of 2,2',4,4'-THBP attached to the bead surface was 0.2 mg per gram of beads, and the amount attached per bead surface area was 0.009 mg per 1 cm ² . 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 2,2',4,4'-THBP was used, a remarkable blood coagulation promoting effect was observed, and the serum separation condition was also very good. Ta. Example 2 2,2',4,4'-THBP0.2g, ellagic acid 0.2
A liquid blood coagulation promoter was prepared from 100 g of water and 100 g of water. After placing 0.1 cc of the above blood coagulation promoter into a polymethyl methacrylate spittoon, 5 cc of blood was collected from a normal healthy person and immediately injected at 20°C.
I left it there. Next, the blood coagulation time was measured in the same manner as in Example 1, and the state of blood separation was observed. As is clear from the results in the Example 2 column of Table 1, when 2,2',4,4'-THBP and ellagic acid were used together, a remarkable synergistic effect was observed in promoting blood coagulation, and serum separation status was observed. It was also very good. Example 3 0.2 g of 2,2',4,4'-THBP, 0.8 g of colloidal silica obtained by reducing silicon tetrachloride in a reducing air flow (average particle size 5 mμ to 40 mμ), 100 g of water
A liquid dispersion of blood coagulation promoter was prepared. After putting 0.1 cc of the above blood coagulation promoter into a polymethyl methacrylate spittoon, 5 cc of blood collected from a normal healthy person was immediately injected, and the temperature was kept at 20°C.
I left it there. 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 in the Example 3 column of Table 1, when 2,2',4,4'-THBP and colloidal silica are used together, a remarkable synergistic effect is observed in promoting blood coagulation, and serum separation status is observed. It was also very good. Comparative Example 1 2,2',4,4' - 1 g of polystyrene beads with a diameter of 2.5 mm to which THBP is not attached, an outer diameter of 15 mm,
A container made of polymethyl methacrylate with a height of 100 mm 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 1 column of Table 1. Comparative Example 2 A liquid blood coagulation promoter was prepared consisting of 0.5 g of ellagic acid and 100 g of water, and 0.1 cc of this was placed in a polymethyl methacrylate spittoon having an outer diameter of 15 mm and a height of 1.00 mm. 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 2 column of Table 1. Comparative Example 3 Colloidal silica (average particle size 5mμ to 40m
A blood coagulation promoter was prepared in the form of a dispersion liquid 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.

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Claims (1)

[Scope of Claims] 1. A blood coagulation promoter characterized by comprising 2,2',4,4'-tetrahydroxybenzophenone. 2. A blood coagulation promoter comprising 1 part by weight of 2,2',4,4'-tetrahydroxybenzophenone and 0.1 to 5.0 parts by weight of ellagic acid. 3. It is characterized by consisting of 1 part by weight of 2,2',4,4'-tetrahydroxybenzophenone and 0.05 to 10 parts by weight of an adsorbent inorganic substance selected from the group consisting of glass, silica, kaolin, celite, and bentonite. Blood coagulation promoter.