JP2004189682A - Antiplatelet action of glycyrrhizin - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a pharmaceutical composition for suppressing accumulation of platelets into the lung and/or the liver and/or degranulation of the platelets. <P>SOLUTION: The pharmaceutical composition comprises glycyrrhizin or glycyrrhetic acid in an active amount for suppressing accumulation of platelets into the lung and/or the liver and/or degranulation of the platelets and arbitrarily a pharmaceutically acceptable carrier. <P>COPYRIGHT: (C)2004,JPO&NCIPI

Description

【００２６】
【発明の効果】
本発明においては、グリチルリチン及びグリチルレチン酸を利用して、肺及び/又は肝臓への血小板の集積、及び/又は当該血小板の脱顆粒を抑制することが可能である。
【図面の簡単な説明】
【図１】この図は、グリチルリチンの投与による抗血小板作用を示すグラフである。[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a pharmaceutical composition for inhibiting platelet accumulation in the lung and / or liver and degranulation of the platelet, and a method for producing the same.
[0002]
Problems to be solved by the prior art and the invention
[0003]
The present inventors intravenously inject lipopolysaccharide (LPS; also called endotoxin), which is a membrane component of Gram-negative bacteria, and bacterial cells (Heat-killed bacteria such as Gram-negative bacteria, Gram-positive bacteria, and fungi) themselves into mice. Then, they found that an immediate platelet reaction occurs, and have already reported it in the literature (Endo Y. & Nakamura M., Br. J. Pharmacol., 1992, 105: 613).
[0004]
[Non-patent document 1]
Endo, Nakamura, "The effect of lipopolysaccharide, interleukin-1, and tumour necrosis factor on the hepatic accumulation of 5-hydroxytryptamine and platelet in the mouse" (The effect of lipopolysaccharide) , interleukin-1, and tumor necrosis factor on the hepatic accumulation of 5-hydroxytryptaine and platelets in the mouse), "British Journal of Pharmacology", (UK), 1992, vol. 105, p613.
[Non-patent document 2]
Kroes, Boekelman, van den Berg, wolbink, van Dijk, Labadie, "Inhibition of Human Completion by Beta- "Glycyrrhetinic acid"("Inhibition of human complement by b-glycyrrhetinic acid"), "Immunology", (UK), 1997, vol. 90, p115-120.
[Non-Patent Document 3]
Shibazaki, Kawabata, Yokochi, Nishida, Takada, Endo, "Complement-Dependent Accumulation and Degradation of Platelets in the Lang and River Induces" , “Complement-dependent accumulation and degradation of platelets in the lung and liver induced following injection of lipopolysaccharides”, “Infection and Immunology”, (USA), 1999, Volume 67, p5186-5191.
[Non-patent document 4]
Zhao, Ohtaki, Yamaguchi, Matsushita, Fujita, Yokochi, Takada, Endo, "El Ps-Indust Platelet Response and Rapid Shock in Mice: Contribution of O-Antigen Region of LPS and Involvement of the Lectin Pathway of the Completion System ("LPS-induced platelet response and rapid shock in mice: contribution of O -antigen region of LPS and involvement of the lectin pathway of the complement system ")," Blood ", (USA), 2002, vol. 100, p3233-3239.
[0005]
When LPS is intravenously administered to mice,
(1) Platelets accumulate in the lungs and liver within 2 to 3 minutes after LPS administration, especially large amounts of platelets accumulate in the lungs,
(2) Five to ten minutes after LPS administration, the return of platelets to the bloodstream begins,
(3) When the dose of LPS is high, platelets accumulated in the liver undergo degranulation, which causes anaphylactic shock,
(4) Ten minutes after LPS administration, platelet accumulation in the spleen occurs, and platelet accumulation in the liver occurs relatively slowly after one to three hours.
Of these reactions, the reactions (1) and (2) are called "immediate platelet reactions" and are used in this sense in this application. That is, a reaction that occurs in a living body within 10 minutes after LPS administration is referred to as such.
[0006]
The present inventors have confirmed by an electron microscope that platelets accumulate in the lung and liver by the immediate platelet reaction, and furthermore, the immediate platelet response was induced by the activation of the complement system lectin pathway. (Shibazaki et al., Infect. Immun., 1999, 67: 5186-5191).
[0007]
The present inventors have reported that glycyrrhizin (GL) and glycyrrhetinic acid in which a sugar chain has been removed from GL have an anti-complement effect in vitro (Kroes et al., Immunology, 1997, 90: 115-120; Zhao et al., Blood, 2002, 100: 3233-3239). An object of the present invention is to provide a pharmaceutical composition for inhibiting platelet accumulation in the lung and liver, degranulation of the platelets, and a method for producing the same.
[0008]
[Means for Solving the Problems]
The present inventors have further studied intensively based on the above-mentioned research results, and as a result, have found that glycyrrhizin and glycyrrhetinic acid have an antiplatelet effect, and have completed the present invention.
[0009]
That is, the present invention is a pharmaceutical composition for inhibiting platelet accumulation in the lung and / or liver, and degranulation of the platelets, and is effective in inhibiting the accumulation and / or the degranulation. There is provided a pharmaceutical composition comprising an amount of glycyrrhizin or glycyrrhetinic acid, optionally together with a pharmaceutically acceptable carrier.
[0010]
The present invention further relates to a method for producing a pharmaceutical composition which suppresses the accumulation of platelets in the lung and / or liver, and the degranulation of the platelets, the method comprising adding glycyrrhizin or glycyrrhetinic acid. provide.
[0011]
Glycyrrhizin (or glycyrrhizic acid) is a compound consisting of glycyrrhizic acid and two molecules of glucuronic acid, which has been known for a long time to have an anti-inflammatory effect. It is also known to have an antiallergic effect, an immunosuppressive activity, a liver function enhancing effect, a detoxifying effect, an effect of increasing resistance to viruses, and the like. Particularly, it is a compound widely used in the clinical field as an agent for liver disease and an agent for allergy.
[0012]
Glycyrrhizin has an inhibitory effect on HIV intracellular proliferation in recent years, and when glycyrrhizin is administered to patients with asymptomatic carriers (AC) at 150 to 225 mg daily, they survive without AIDS for more than 10 years It is reported to have an effect.
[0013]
BEST MODE FOR CARRYING OUT THE INVENTION
Glycyrrhizin and glycyrrhetinic acid used in the present invention can be, for example, those available from Minophagen Pharmaceutical. Glycyrrhizin and glycyrrhetinic acid used in the present invention can also be obtained by extracting from licorice. Pharmaceutically acceptable salts of glycyrrhizin and glycyrrhetinic acid can also be used, but examples of such salts include those obtained by reacting glycyrrhizin with an inorganic or organic base at a fixed molar ratio. Glycyrrhizin ammonium salt, glycyrrhizin alkali metal salt, or glycyrrhizin choline salt. In the present invention, one kind of glycyrrhizin or a salt thereof can be used, and a mixture of any two kinds of salts can be used. It is preferred to use one of the salts.
[0014]
In using the pharmaceutical composition containing glycyrrhizin and glycyrrhetinic acid of the present invention, a pharmaceutical composition formulated by appropriately blending glycyrrhizin and glycyrrhetinic acid as active ingredients in a pharmaceutically acceptable carrier or the like is used. It may be prepared, and the preparation may be performed according to a method known in the art. In the present invention, such a pharmaceutical composition can be suitably used.
[0015]
The pharmaceutical composition can be prepared in various forms according to a conventional method. The dosage form is not particularly limited, but one or more pharmaceutically acceptable vehicles, carriers, excipients, binders, disintegrants, dispersants, lubricants, coatings, preservatives, stable By mixing with agents, flavors / flavors, coloring agents, etc., tablets, powders, granules, capsules, liquid medicines and other internal preparations, and sterile solutions, suspensions and other injections can be prepared. These can be manufactured using a known technique.
[0016]
As for the dosage form, for example, glycyrrhizin or a pharmacologically acceptable salt thereof, a binder such as corn starch and gelatin, a shaper such as microcrystalline cellulose, a bulking agent such as sodium alginate, a sweetener such as lactose, sucrose, etc. By dispensing ingredients and the like, powders, tablets, granules, and capsules can be appropriately prepared. These dosage forms are used for oral administration. When used as an injection, distilled water for injection or polyethylene glycol is used as the solution, or a dispersing agent, a buffer, a preservative, an antioxidant, etc. may be added as necessary. You can also. Further, glycyrrhizin or a pharmacologically acceptable salt thereof, an absorption enhancer, a pH adjuster, and if necessary, ursodesoxycholic acid and the like can be blended to prepare a rectal preparation.
[0017]
Specific applications of the pharmaceutical composition used in the present invention include, for example, adult respiratory insufficiency caused by bacterial infection (gram-negative bacteria, gram-positive bacteria, and fungus-induced sepsis). Includes mammalian diseases. In addition, it can be applied not only to diseases caused by endotoxin circulating in the circulatory system but also to inflammation in local bacterial infections such as skin infections, because platelets are involved in such inflammations. It is possible. In addition, the present invention can be applied to general inflammation due to infection.
[0018]
The pharmaceutical composition used in the present invention can be administered to mammals by oral, enteral, topical, transdermal, intravascular, intramuscular administration and the like. It is appropriately selected by a clinician depending on the weight and health of the patient to be treated or prevented, and the condition of the disease to be treated.
[0019]
When the antiplatelet agent is administered to the subject in the use of the present invention, it is usually administered at a dosage of 0.1 to 100 mg per kg of body weight per day.
[0020]
【Example】
Example 1 Influence of Glycyrrhizin and Glycyrrhetinic Acid on Immediate Platelet Response The accumulation of platelets in the lung was measured using serotonin (5HT) contained in a large amount in platelets as an index. The experiments were performed at room temperature (26-28 ° C.) (because the platelet response to LPS depends on room temperature). BALB / c mice were intravenously injected with glycyrrhizin (200 mg / kg) prepared in physiological saline or physiological saline. Ten minutes after the injection, mice were intravenously injected with LPS (2 mg / kg) derived from Klebsiella O3 bacteria (KO3). At 4 minutes and 4 minutes after LPS administration, the mice were decapitated, and the blood flowing out of each mouse was first used for platelet count measurement (2-3 drops) and then for serotonin measurement. (1-2 drops). Immediately after blood collection, lungs were removed and stored with dry ice until serotonin was measured. The platelet count was measured by an automatic blood cell analyzer. Serotonin concentrations in blood and lung were measured by a fluorescence method after extracting serotonin with perchloric acid, separating the extract by column chromatography using an ion exchanger. Serotonin in the blood was unstable, so the measurement was performed immediately after blood collection. The measurement was carried out according to the method described in the previous report (Yoshida A. et al., Br. J. Pharmacol. 2002; 137: 146-152). The degree of shock was indicated by the following score observed from 5 to 10 minutes after LPS administration. Each measured value was shown as an average value +/- SD (n = 4). In the experiment shown in FIG. 1, a total of 20 mice were used. Prior to this experiment, the effect of glycyrrhizin itself (200 mg / kg) on mouse platelets was examined, but no significant difference was found.
Score indicating the degree of shock:
0: No symptoms 1: Difficulty walking 2: Stomaching 3: Weak convulsions 4: Strong convulsions
[0021]
Results The results are shown in FIG. FIG. 1 (a) is a graph showing the platelet count in blood. It can be seen that 4 minutes after LPS administration, the control group of mice administered with physiological saline had a significantly reduced platelet count. On the other hand, in the mice in the treatment group to which glycyrrhizin was administered, the platelet count was decreased as in the control group, but the degree was smaller than that in the control group. Ten minutes after LPS administration, the mice in the treatment group and the control group both had recovered blood platelet counts, but the degree of recovery (the slope of the straight line in the graph) was greater in the treatment group administered with glycyrrhizin. It turns out that it is big. In addition, the platelet count in blood at both 4 minutes and 10 minutes after LPS administration had a statistically significant difference between the treated group and the control group (p <0.05).
[0022]
FIG. 1 (b) shows a comparison of the serotonin concentration in the blood, and a result corresponding to the result of the experiment on the number of platelets was obtained. The medium concentration was decreased in both the treatment group and the control group, but the degree of decrease was lower in the treatment group, and the serotonin concentration was recovered 10 minutes after LPS administration.
[0023]
FIG. 1 (c) shows the result of measuring the serotonin concentration in the lung. In contrast to the change in the serotonin concentration in the blood, the serotonin concentration in the lung increased 4 minutes after LPS administration. The degree of the increase was greater in the control group, and in the mice in the treatment group to which glycyrrhizin was administered, the increase in serotonin concentration was lower than that in the control group. At 10 minutes after LPS administration, an increase in serotonin concentration in the lung was suppressed, and a tendency for recovery was observed. Serotonin levels at both 4 and 10 minutes after LPS administration were statistically significant between the treated and control groups (p <0.05).
[0024]
Table 1 shows the results of comparing the degree of shock of mice after LPS administration between the treatment group and the control group. As shown, the mean score of shock for mice in the treatment group was 2.00 +/- 0.4 (mean +/- SD), while the mean score for shock in mice in the control group was , 3.75 +/- 0.5 (Id.), Which were significant differences.
[0025]
[Table 1]

[0026]
【The invention's effect】
In the present invention, glycyrrhizin and glycyrrhetinic acid can be used to suppress platelet accumulation in the lung and / or liver and / or degranulation of the platelets.
[Brief description of the drawings]
FIG. 1 is a graph showing an antiplatelet effect by administration of glycyrrhizin.

Claims (2)

A pharmaceutical composition for inhibiting accumulation of platelets in the lung and / or liver, and degranulation of the platelets,
A pharmaceutical composition comprising an effective amount of glycyrrhizin or glycyrrhetinic acid to inhibit the accumulation and / or the degranulation, optionally together with a pharmaceutically acceptable carrier. A method for producing a pharmaceutical composition which suppresses accumulation of platelets in the lung and / or liver and degranulation of the platelets, comprising glycyrrhizin or glycyrrhetinic acid.

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