JPH0126025B2 - - 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. The present invention relates to a blood coagulation promoter that is mixed into collected blood for purposes that can be accomplished in a short period of time. 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 sample. 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, bentonite, etc. to accelerate coagulation, a method of adding small spheres or berets 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. 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, certain phosphite esters have been found to have an effective effect on factor XII and platelets in blood coagulation, such as kaolin, celite, and
It has been discovered that it has a more excellent activity than glass, ellagic acid, etc. In addition, the combined use of such phosphites with 2,2',4,4'-tetrahydroxybenzophenone, catechol derivatives, P,P'-isopropylidene diphenol, and adsorbent inorganic substances can synergistically improve blood flow. We have discovered that it exhibits a coagulation-promoting effect. The gist of the present invention is as follows: 1 formula (In the above formula, each of R ₁ , R ₂ , and R ₃ is a phenyl group, or 1 to 3 of the hydrogen atoms of the phenyl group
is an alkyl phenyl group substituted with a nonyl group or an octyl group, R ₁ , R ₂ ,
One or two of R ₃ is a phenyl group, or an alkylphenyl group in which 1 to 3 of the hydrogen atoms of the phenyl group are substituted with a nonyl group or an octyl group, and the other is a decyl group. ) A blood coagulation promoter, characterized in that it is composed of a phosphite ester represented by the formula 2, and is used in an amount of 0.01 mg to 10 mg per 5 ml of blood volume. (In the above formula, each of R ₁ , R ₂ , and R ₃ is a phenyl group, or 1 to 3 of the hydrogen atoms of the phenyl group
is an alkylphenyl group substituted with a nonyl group or an octyl group, R ₁ , R ₂ ,
One or two of R ₃ is a phenyl group, or an alkylphenyl group in which 1 to 3 of the hydrogen atoms of the phenyl group are substituted with a nonyl group or an octyl group, and the other is a decyl group. ) 1 part by weight of a phosphite ester represented by
A blood coagulation promoter comprising 0.1 to 3.0 parts by weight of 2,2',4,4'-tetrahydroxybenzophenone, and characterized in that phosphite is used in an amount of 0.01 mg to 10 mg per 5 ml of blood volume. , 3 formulas (In the above formula, each of R ₁ , R ₂ , and R ₃ is a phenyl group, or 1 to 3 of the hydrogen atoms of the phenyl group
is an alkyl phenyl group substituted with a nonyl group or an octyl group, R ₁ , R ₂ ,
One or two of R ₃ is a phenyl group, or an alkylphenyl group in which 1 to 3 of the hydrogen atoms of the phenyl group are substituted with a nonyl group or an octyl group, and the other is a decyl group. ) 1 part by weight of phosphite ester represented by
- 0.1 to 5.0 parts by weight of a catechol derivative selected from tertiary butylcatechol, 4-tertiarybutylcatechol, paratertiarybutylcatechol, catechin, and nordihydrogayaretic acid, and phosphite per 5 ml of blood volume. A blood coagulation promoter, characterized in that it is used in an amount of 0.01mg to 10mg, formula 4. (In the above formula, each of R ₁ , R ₂ , and R ₃ is a phenyl group, or 1 to 3 of the hydrogen atoms of the phenyl group
There is an alkyl phenyl group in which 3 are substituted with a nonyl group or an octyl group, or one or two of R ₁ , R ₂ , and R ₃ are phenyl groups, or 1 to 3 of the hydrogen atoms of the phenyl group is an alkyl phenyl group substituted with a nonyl group or an octyl group, and the others are decyl groups. ) 1 part by weight of phosphite ester represented by P,
P′-isopropylidene diphenol 0.1 to 5.0
A blood coagulation promoter, characterized in that the phosphite ester is used in an amount of 0.01 mg to 10 mg per 5 ml of blood volume. (In the above formula, each of R ₁ , R ₂ , and R ₃ is a phenyl group, or 1 to 3 of the hydrogen atoms of the phenyl group
is an alkyl phenyl group substituted with a nonyl group or an octyl group, R ₁ , R ₂ ,
One or two of R ₃ is a phenyl group, or an alkylphenyl group in which 1 to 3 of the hydrogen atoms of the phenyl group are substituted with a nonyl group or an octyl group, and the other is a decyl group. ) and 0.05 to 10 parts by weight of an adsorbent inorganic material selected from glass, silica, kaolin, celite, and bentonite with a particle size of 50μ or less, and the phosphite is A blood coagulation promoter, characterized in that it is used in an amount of 0.01 mg to 10 mg per 5 ml. 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 phosphite represented by the following formula. (In the above formula, R ₁ , R ₂ , and R ₃ are all phenyl groups, or alkylphenyl groups in which 1 to 3 hydrogen atoms of the phenyl group are substituted with nonyl groups or octyl groups. , one or two of R ₁ , R ₂ , and R ₃ is a phenyl group, or an alkylphenyl group in which 1 to 3 hydrogen atoms of the phenyl group are substituted with a nonyl group or an octyl group, and others is a decyl group.) In the case of the above formula, the phosphite is R ₁ ,
They are classified as follows depending on the type of R ₂ and R ₃ . (1) When R ₁ , R ₂ , and R ₃ are all phenyl or alkylphenyl groups: For example, trisphenyl phosphite, tris (monononylphenyl) phosphite, tris (dinonylphenyl) phosphite, tris (torino These include nilphenyl) phosphite, tris(monoctylphenyl) phosphite, tris(dioctyl phenyl) phosphite, tris(trioctylphenyl) phosphite, and the like. (2) When R ₁ and R ₂ are a phenyl group or an alkylphenyl group and R ₃ is a decyl group: For example, diphenyldecyl phosphite, di(monononylphenyl)decyl phosphite,
di(dinonylphenyl)decyl phosphite,
Examples include di(trinonylphenyl)decyl phosphite, di(monooctyl phenyl) decyl phosphite, di(dioctyl phenyl) decyl phosphite, and di(trioctylphenyl) decyl phosphite. (3) When R ₁ is a phenyl group or an alkylphenyl group and R ₂ and R ₃ are decyl groups: For example, phenyl didecyl phosphite, nonylphenyl didecyl phosphite, dinonylphenyl didecyl Examples include phosphite, trinonylphenyl didecyl phosphite, monooctylphenyl didecyl phosphite, dioctyl phenyl didecyl phosphite, trioctyl phenyl didecyl phosphite, and the like. Among the above phosphite esters, those particularly preferably used include tris(monononylphenyl) phosphite, tris(dinonylphenyl) phosphite, tris(trinonylphenyl) phosphite, diphenyldecyl phosphite, phenyldidecyl phosphite and the like. As the blood coagulation promoter in the present invention, the above-mentioned phosphite can be directly added to blood. 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, varafin, chlorinated varafin, chlorinated polyethylene, chlorinated polybutene,
Polyethylene glycol, cellulose, cellulose acetate, chitin, etc. are used. 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; and glycerin. Esterified products of polyhydric alcohols such as 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 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 polymeric substance to migrate to the surface of the molded globules or pellets and to effectively promote blood coagulation, it is necessary to have the necessary amount present on the surface. It is especially preferred 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 use it by dissolving or dispersing it in ethylene glycol, glycerin, etc., and high-viscosity liquids such as polyethyl acrylate oligomers, liquid polybutadiene, liquid paraffin, chlorinated paraffin, chlorinated polybutene, polyethylene glycol, and other polymer oligomers. It can be used by mixing it with oil such as glycerin or silicone oil. In the present invention, the effective amount of phosphite added to blood is 0.01 mg to 10 mg per 5 ml of blood volume.
It is. The blood coagulation promoter of the present invention exhibits a significant blood coagulation promoting effect when the above-mentioned phosphite is used alone. However, the above phosphite ester and 2,
2',4,4'-tetrahydroxybenzophenone,
It was found that when catechol derivatives, p,p'-isopropylidene diphenol, and adsorbent inorganic substances were used together, it had a more excellent blood coagulation promoting effect than when the above-mentioned phosphite was used alone. . One of the substances that exhibits a synergistic blood coagulation promoting effect when used in combination with the above-mentioned phosphite is 2,2',4,4'-tetrahydroxybenzophenone. 2,2',4,4'-tetrahydroxybenzophenone has the following chemical structural formula. 2,2',4,4'-tetrahydroxybenzophenone has a melting point of 195℃ and a solubility in solvents of 30℃.
0.1% by weight for water, 50% by weight for methanol, 40% by weight for ethanol, and 10% by weight for a 1:1 water-ethanol solution. Further, the maximum absorption wavelength position is 345 mμ, and Color Parieux (Gardner) is No. 8 in a 1% by weight methanol solution. And 2, 2', 4,
It has been known that 4'-tetrahydroxybenzophenone can be incorporated into synthetic resins, cosmetics, etc. as an ultraviolet absorber, but its applicability as a blood coagulation promoter was completely unknown. The weight ratio of 2,2',4,4'-tetrahydroxybenzophenone is 1 part phosphite.
0.1 to 3.0 are used. Other substances that synergistically exhibit a blood coagulation promoting effect when used in combination with the phosphite are catechol derivatives. As the catechol derivative, one represented by the following formula is used. In the case of the above formula, the catechol derivative is R ₁ ,
Depending on the type of R ₂ and R ₃ , 3-alkylcatechol,
It is divided into 4-alkylcatechol, para-alkylcatechol, catechin, nordihydrogayaretic acid, ellagic acid, etc. Among these, those which show particularly remarkable synergistic effects are 3-tert-butylcatechol, 4-tert-butylcatechol, para-tert-butylcatechol, catechin, and nordihydrogayaretic acid. The weight ratio of the catechol derivative used is 0.1 to 5.0 to 1 part of the phosphite ester. Another substance that synergistically exhibits a blood coagulation promoting effect when used in combination with the phosphite is p,p'-isopropylidene diphenol.
This is a substance commercialized as bisphenol A and has been used as a raw material for producing epoxy resins, but it was not known to have a blood coagulation promoting effect. The weight ratio of p,p'-isopropylidene diphenol used is 0.1 to 5.0 per 1 part of the phosphite. Still another substance that synergistically exhibits a blood coagulation promoting effect when used in combination with the phosphite is an adsorbent inorganic substance. The adsorbent inorganic material is selected from glass, silica, kaolin, celite, and bentonite, and these have excellent blood adsorption properties. The particle size of these adsorptive inorganic substances is selected to be 50μ or less. These substances have also been used as blood coagulation promoters, but when used in combination with the above-mentioned phosphite ester, they have a more excellent blood coagulation promoting effect than when each of these substances is used alone. It was confirmed that the When the adsorptive inorganic material is in powder form, the weight ratio of the adsorbent inorganic material used is 0.05 to 10 parts of the phosphorous acid ester. Furthermore, when the adsorbent inorganic substance is in the form of small spheres or pellets, it is particularly preferable to use a substance coated with the phosphite ester melted by heating or dissolved in a solvent. When using the blood coagulation accelerator of the present invention, the blood may be collected into a blood test container such as a spittoon in advance after it has been placed in the container, 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 contamination of residual fibrin and blood clot components in the collected serum is also solved, and furthermore, since the contraction of the blood clot components progresses sufficiently, the yield of serum is significantly increased. Examples of the present invention are described below. Example 1 A blood coagulation promoter was prepared by attaching tris(dinonylphenyl) phosphite to the surface of polystyrene beads having a diameter of 2.5 mm. Tris(dinonylphenyl) phosphite was attached to the surface of the beads by adding an ethanol solution of tris(dinonylphenyl) phosphite to the beads, stirring thoroughly, and then drying the ethanol. The amount of tris(dinonylphenyl)phosphite attached to the surface of the beads was 0.21 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 tris(dinonylphenyl) phosphite was used, a remarkable blood coagulation promoting effect was achieved, and the serum separation condition was also extremely good. Examples 2-3 In Example 1, tris(dinonylphenyl)phosphite was added per gram of polystyrene beads.
Instead of using 0.21 mg of diphenyldecyl phosphite attached to each 1 g of polystyrene beads (Example 2),
Polystyrene beads in which 0.22 mg of phenyldidecyl phosphite was attached per gram of polystyrene beads (Example 3) were 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. As is clear from the results of Examples 2 and 3 in Table 1, a remarkable blood coagulation promoting effect was obtained in both cases, and the serum separation state was also extremely good. Examples 4 to 7 0.12 mg of tris(dinonylphenyl) phosphite and 0.10 mg of 2,2',4,4'-tetrahydroxybenzophenone were attached to the surface of polystyrene beads with a diameter of 2.5 mm per 1 g of beads (example). Example 4), 0.11 mg of tris(dinonylphenyl) phosphite per 1 g of polystyrene beads and 4
- 0.08 mg of tertiary butylcatechol attached (Example 5), 0.11 tris(dinonylphenyl) phosphite per 1 g of polystyrene beads
mg and p,p'-isopropylidene diphenol
0.11 mg of Tris(dinonylphenyl) per 1 g of polystyrene beads (Example 6)
A sample to which 0.11 mg of phosphite and 0.15 mg of fine silica powder with a particle size of 2 to 5 μm were attached (Example 7) was prepared. For attachment to the bead surface, a solution in which the attachment component is dissolved or suspended in ethanol is added to the beads,
This was done by thoroughly stirring and then drying the ethanol. 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 4 to 7 in Table 1, a remarkable blood coagulation promoting effect was obtained in all cases, and the serum separation state 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 5mμ to 40m
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】

Claims (1)

[Claims] 1 formula (In the above formula, R ₁ , R ₂ , and R ₃ are all phenyl groups, or alkylphenyl groups in which 1 to 3 hydrogen atoms of the phenyl group are substituted with nonyl groups or octyl groups. , one or two of R ₁ , R ₂ , and R ₃ is a phenyl group, or an alkylphenyl group in which 1 to 3 hydrogen atoms of the phenyl group are substituted with a nonyl group or an octyl group, and others is a decyl group), and is used in an amount of 0.01 mg to 10 mg per 5 ml of blood volume. 2 formulas (In the above formula, R ₁ , R ₂ , and R ₃ are all phenyl groups, or alkylphenyl groups in which 1 to 3 hydrogen atoms of the phenyl group are substituted with nonyl groups or octyl groups. , one or two of R ₁ , R ₂ , and R ₃ is a phenyl group, or an alkylphenyl group in which 1 to 3 hydrogen atoms of the phenyl group are substituted with a nonyl group or an octyl group, and others is a decyl group.) 1 part by weight of a phosphite ester represented by
2',4,4'-tetrahydroxybenzophenone 0.1
3.0 parts by weight, and the phosphite ester is used in an amount of 0.01 mg to 10 mg per 5 ml of blood volume. 3 formulas (In the above formula, R ₁ , R ₂ , and R ₃ are all phenyl groups, or alkylphenyl groups in which 1 to 3 hydrogen atoms of the phenyl group are substituted with nonyl groups or octyl groups. , one or two of R ₁ , R ₂ , and R ₃ is a phenyl group, or an alkylphenyl group in which 1 to 3 hydrogen atoms of the phenyl group are substituted with a nonyl group or an octyl group, and others is a decyl group.) 1 part by weight of a phosphite ester represented by
It consists of 0.1 to 5.0 parts by weight of a catechol derivative selected from tertiary butylcatechol, 4-tertiarybutylcatechol, paratertiarybutylcatechol, catechin, and nordihydrogayaretic acid, and phosphite per 5ml of blood volume.
A blood coagulation promoter characterized in that it is used in an amount of 0.01mg to 10mg. 4 formula (In the above formula, R ₁ , R ₂ , and R ₃ are all phenyl groups, or alkylphenyl groups in which 1 to 3 hydrogen atoms of the phenyl group are substituted with nonyl groups or octyl groups. , one or two of R ₁ , R ₂ , and R ₃ is a phenyl group, or an alkylphenyl group in which 1 to 3 hydrogen atoms of the phenyl group are substituted with a nonyl group or an octyl group, and others is a decyl group) and 0.1 to 5.0 parts by weight of P,P'-isopropylidene diphenol, and 0.01 mg to 10 mg of phosphite is used per 5 ml of blood volume. A blood coagulation promoter characterized by: 5 formula (In the above formula, R ₁ , R ₂ , and R ₃ are all phenyl groups, or alkylphenyl groups in which 1 to 3 hydrogen atoms of the phenyl group are substituted with nonyl groups or octyl groups. , one or two of R ₁ , R ₂ , and R ₃ is a phenyl group, or an alkylphenyl group in which 1 to 3 hydrogen atoms of the phenyl group are substituted with a nonyl group or an octyl group, and others is a decyl group), and 0.05 to 10 parts by weight of an adsorbent inorganic substance selected from glass, silica, kaolin, celite, and bentonite and having a particle size of 50μ or less, A blood coagulation promoter characterized in that 0.01 mg to 10 mg of phosphite is used per 5 ml of blood volume.