JP2005213156A - Crude drug-containing composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a composition which prevents and improves the inhibition of immunocompetent cells due to being under stress, particularly the inhibition of the cellar immunity function. <P>SOLUTION: The composition comprises muira-puama having an action of appropriately maintaining the blood glucocorticoid level which rises on loading stress and a crude drug having an action of accelerating the production of interferon γ from splenic cells in the presence of a glucocorticoid, specifically one or more kinds selected from the dried seed of Helianthus annuus L. in Compositae, the dried root skin of Lycium chinense Miller in Solanaceae, the dried ripened pollen of Typha angustifolia L., T. latifolia L., T. orientalis Presl., T. davidiana Hand.-Mazz. or T. minima Funk. in Typhaceae, the dried root and rhizome of Aster tataricus L.f. in Compositae, the dried style and stigma of new flowers of Zea mays L. in Gramineae, the dried, exodermis-removed tuber of Sparganium erectum L. subsp. Stoloniferum Hara, S. simplex Huds. or S. Stenophyllum Maxim. in Sparganiaceae, the dried tuber of Arisaema consanguineum Scott, A. amurense Maxim., A. heterophyllum Blume, A. ambignum Engler, A. japonicum Blume var. atropureum (Engler) Kitamura or A. peninsulae Nakai in Araceae, and the crude drug made from Achillea millefolium L. in Compositae. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a drug effective for prevention or improvement of susceptibility to various viruses, which is a major factor of infection caused by immunosuppression caused by stress.

  Currently, modern people living in a complex social structure are exposed to a lot of stress. When humans are stressed, reactions such as an increase in blood pressure, an increase in body temperature, and an increase in blood sugar level occur. This is a reaction of the living body (stress reaction) in order to recover the abnormality of homeostasis (homeostasis) due to stress. This stress response is regulated by stress hormones such as glucocorticoids.

  When stress is applied for a long time, the blood glucocorticoid concentration continues to be high. Under such circumstances, adverse effects are caused in maintaining the homeostasis of the living body.

  The effect is particularly great in the immune system. Suppression of immunocompetent cells is caused by an increase in blood corticocoid concentration caused by stress. In particular, suppression of cellular immune function is remarkable. This cellular immune function is known to play an important role in resistance to viral infections that cause infections such as the common cold and influenza. Currently, there are no known readily available drugs that directly kill various viruses. Therefore, it can be said that the improvement of the body defense function, especially the activation of cellular immunity, which plays an important role in virus elimination, is the only effective treatment / prevention method.

A technique has been disclosed in which muirapuama regulates in vivo cortisol concentration under stress (Patent Document 1).
Japanese Patent Laid-Open No. 2003-40789

  An object of this invention is to provide the composition which prevents and improves suppression of the immunocompetent cell by receiving stress, especially suppression of cellular immune function. That is, an object of the present invention is to provide a composition that activates cellular immunity that is important for improving the defense function of a living body under stress load, particularly for virus elimination.

  As a result of diligent research to solve the above-mentioned problems, the present inventors have found that muirapuama, which has an effect of appropriately maintaining the blood glucocorticoid concentration that rises during stress loading, and interferon γ (hereinafter referred to as spleen cells) in the presence of glucocorticoid. Abbreviated to “INFγ”) containing a herbal medicine group having a production promoting action, specifically, one or more selected from Kojitsukiki, Jikoppi, Hou, Zion, Gyobabeju, Kaisan Ryo, Tennansho, and Yodo It has been found that the characteristic composition has an action to improve the stress-susceptible state, and the present invention has been completed.

  In other words, the present invention significantly improves the stress-susceptible state by combining muirapuama with a herbal medicine having an INFγ production promoting action. Patent Document 1 discloses a cold and influenza therapeutic agent containing muirapuama as an active ingredient, but the present invention further has a remarkably excellent effect by adding a crude drug having an INFγ production promoting action.

  According to the present invention, against the virus that causes infection caused by corticosterone that is excessively secreted by stress, through two mechanisms of suppression of excessive secretion of corticosterone or immune activation in an immunosuppressed state. It became possible to prevent and treat easily infectious diseases.

  The muirapuama used in the present invention is a medicinal plant based on the root of Ptychopetalum olacoides, Ptychopetalum uncinatum or Liriosma ovata of the Olacaceae plant that grows in the Amazon River basin of Brazil. It has been used empirically and in the private sector in the hope of acting on the reproductive and circulatory systems. In particular, muirapuama liquor and its decoction have been used mainly in anticipation of aphrodisiac, tonic, and sperm action, as well as effects on the central nervous system, improvement of paralysis and motor dysfunction, and action on the digestive system. In the literature, such as indigestion, loss of appetite, stomach weakness, diarrhea, dysentery, duodenal helminthiasis and the like are described in the literature.

  Zicopi used in the present invention is a dried root bark of a medicinal plant based on Lycium chinense Miller, a solanaceae plant, and has been used as an antipyretic and tonic and has a known antihypertensive effect. It has been.

  The koji tsushiki used in the present invention is a dried seed of a medicinal plant based on the compositae plant Kelitsuki Helianthus annuus L. and has been used in the private sector as a nourishing tonic and antipruritic agent.

  As for the cormorant used in the present invention, Typha angustifolia L., Gama T. latifolia L., Kogama T. orientalis Presl., And others T. davidiana Hand.-Mazz., T. minima Funk. The dried medicinal pollen of a medicinal plant as a base, which has been used in anticipation of the diuretic action by the contained flavonoid and the hemostatic effect by the vasoconstriction action.

  Zion used in the present invention is a medicinal plant based on the compositae Zion Aster tataricus L. f., Which has been thoroughly washed and dried, and has been used as an antitussive, expectorant, diuretic. It was.

  Gyobe beige used in the present invention is a dried medicinal plant and stigma of a medicinal plant based on Gramineae corn Zea mays L. It has been reported to have diuresis, hypotension, and promotion of bile secretion. ing.

  As for the Kasan Ryo used in the present invention, a tuber of a medicinal plant based on Sparganium erectum L. subsp. Stoloniferum Hara, Ezomikuri S. simplex Huds. And Shimenostea stenophyllum Maxim. It has been peeled and dried, and has been used for shakuju, menopause, postpartum nausea, and milk.

  The Tennansho used in the present invention is Arisaema consanguineum Shott, A. amurense Maxim., A. heterophyllum Blume, A. ambignum Engler, A. japonicum Blume var. Atropureum. (Engler) Kitamura, A. peninsulae Nakai, and other tubers of the genus Arisaema are used as they are and dried by cutting them into slices and used as antispasmodic, expectorant and dehumidifying agents. In the private sector, powder has been applied to wounds for the purpose of analgesia and antitumor.

  Pteridocarpus used in the present invention is a herbal medicine made from a medicinal plant based on the compositae plant Achillea millefolium L., which is hemostatic, anti-inflammatory, bitter healthy stomach, blood pressure drop and slight heat loss. The action is known.

The herbal medicine used in the present invention including muirapuama can use an extract in addition to the herbal powder. Here, the extract can be in any form such as a herbal extract, a concentrated extract, and a dry extract. The extract can be produced by an ordinary method using water, alcohol (methanol, ethanol, propanol, isopropanol, etc.), ethyl acetate, acetone, a mixed solution thereof or the like as an extraction solvent.
The effective dose of muirapuama in the composition of the present invention can be appropriately increased or decreased in consideration of age, sex, etc., but is usually 10 mg to 5000 mg, preferably 50 mg to 1500 mg per day as an active ingredient equivalent in adults.

  In addition, the effective dose of the crude drug group having an INFγ production promoting effect can be appropriately increased or decreased in consideration of age, gender, etc., but is usually an adult as a crude drug equivalent amount, and in the case of dicoppi, 1 mg to 10000 mg per day, Preferably, it is 5 mg to 2000 mg. In the case of Koujitsuki, the daily dose is 10 mg to 50000 mg, preferably 50 mg to 10000 mg. In the case of cormorant, it is 100 mg to 100,000 mg per day, preferably 500 mg to 10000 mg. In the case of Zion, the daily dose is 100 mg to 100,000 mg, preferably 500 mg to 10000 mg. In the case of Gyokubeige, the daily dose is 10 mg to 50000 mg, preferably 50 mg to 10000 mg. In the case of Keisan Ryo, the daily dose is 10 mg to 50000 mg, preferably 50 mg to 10000 mg. In the case of tennan show, it is 100 mg to 100,000 mg per day, preferably 500 mg to 10000 mg. In the case of Hemophila, it is 10 mg to 50000 mg per day, preferably 50 mg to 10000 mg.

  The present invention includes vitamins, xanthine derivatives, crude drugs, natural products, excipients, pH adjusting agents, cooling agents, suspending agents, antifoaming agents, viscosities, in a qualitative and quantitative range that does not impair the effects of the invention. Thickeners, solubilizers, disintegrants, binders, lubricants, antioxidants, coating agents, colorants, flavoring agents, surfactants, plasticizers, fragrances, etc. can be mixed. Oral or parenteral preparations such as liquids, tablets, granules, powders, capsules, dry syrups, chewable tablets and transmucosal agents can be used.

  The composition of the present invention restores the immunity that decreases during stress to the original state, and has the effect of preventing or treating diseases. In particular, it is effective for the prevention and treatment of infectious diseases caused by viruses and bacteria, and is particularly effective for the prevention and treatment of common cold and influenza.

Hereinafter, the present invention will be described in more detail with reference to test examples.
(Test Example 1)
Constrained stress loading experiments on mice significantly increase blood corticosterone concentration. Therefore, by measuring the blood corticosterone concentration of the mouse administered with the sample containing the test component, it is possible to evaluate the component having an improvement effect on restraint stress. In this study, we confirmed the inhibitory effect of muirapuama on the rise of blood corticosterone concentration during stress loading.

  As test animals, 8-week-old male C3B6F1 mice were used. The mice were stressed for 14 hours at night (18:30 to 8:30) using a mouse restraint cage. The period of stress load was fasting and fasting.

  As a test sample, muirapuama extract was added to a 0.2% Carboxy methyl cellulose (CMC) solution at a ratio of 4000 mg / 10 ml and uniformly stirred. A 0.2% CMC solution was used as a control sample. It was orally administered with a syringe for oral administration once a day for 7 days every day from 7 days before stress loading so that the amount was 0.25 ml / head. In order to measure the corticosterone concentration in blood, blood was collected under anesthesia immediately before stress loading and at 2, 6, and 14 hours after starting stress. In addition, changes in blood corticosterone with time under no stress load were also measured in the same manner.

The results are shown in FIG. At 14 hours after stress loading, the blood corticosterone concentration in the control sample administration group was significantly higher than that in the group not stressed. On the other hand, in the test sample 1 administration group, an increase in blood corticosterone concentration was significantly suppressed as compared with the control sample 1 administration group.
(Test Example 2)
In order to evaluate a crude drug having an activation effect on the cellular immunosuppressive state due to stress, the immune cell activation effect in the presence of corticosterone was measured using mouse spleen cells.
Seven-week-old male C3B6F1 mice were used as test animals. The mouse spleen was removed under anesthesia, a spleen lymphocyte suspension was prepared with the culture solution, and the cell concentration was adjusted to 3 × 10 ⁶ cells / ml.
As test samples, Koji Tsukiki, Dikoppi, Hou, Zion, Ginkgo Beige, Keisan Ryo, Tennansho, and Akishima, each 50% ethanol extract was diluted to 100 mg / mL with DMSO, and further 5 mg / mL in cell culture solution. did. A control sample prepared by diluting DMSO, the solvent used in the test sample, 20-fold with a culture solution was prepared.

  Add each test sample to the culture solution to 100 ng / ml and incubate for 1 hour in an incubator. Then, reconstitute stress and corticosterone for cell stress and Conacanavalin (ConA) for cell stimulation at 50 nM and 5 mg / ml, respectively. In addition, after further incubation for 20 hours, the supernatant was collected and the IFN-γ concentration was measured with an ELISA kit (Pharmingen, CA, USA). A similar experiment was performed for the control sample.

  The results are shown in FIG. The test sample to which each herbal medicine was added significantly increased the IFN-γ concentration in the supernatant compared to the control.

(Reference example)
Listeria monocytogenes is classified as facultative intracellular parasitic bacteria, and induction of cellular immunity is essential for infection healing. Therefore, it can be used as an indicator of the degree of infection of Listeria monocytogenes when examining the degree of cellular immunity activity that plays an important role in virus infection. In addition, since Listeria monocytogenes is easy to handle compared to viruses, it is also effective in simplifying the operation. Therefore, in order to confirm the relationship between increased blood corticosterone concentration during stress and immunosuppression using Listeria monocytogenes, the following experiment was conducted.

  During 14 hours at night (18:30 to 8:30 the following), which was the activity time of mice, stress was applied for 3 consecutive days using a restraint cage for mice. Immediately after the stress load on the first day, Listeria monocytogenes was inoculated intraperitoneally, and the number of Listeria monocytogenes infections in the liver on the third day after infection was measured. The period of stress load was fasting and fasting. The non-stress group was similarly tested.

  The same test was conducted in the group in which RU486, a corticosteroid receptor antagonist, was suspended in Macrogol 400 at a concentration of 25 mg / 10 ml, and 50 ml / head was inoculated subcutaneously immediately before stress loading for 3 consecutive days. It was.

  FIG. 3 shows the results of the reference example. In statistical processing, the average value of the comparison target was tested by Student t-test, and p <0.05 was considered significant.

  The number of Listeria monocytogenes infections in the stress group was significantly increased compared with the non-stress group. On the other hand, the number of Listeria monocytogenes in the RU486-inoculated group was significantly reduced compared to the stress-loaded group. This confirmed that it was closely related to immunosuppression due to increased blood corticosterone concentration due to stress loading. From this result, it is possible to evaluate a drug having an improvement effect on the susceptibility to infection in the state of suppression of cellular immunity due to stress load.

(Test Example 3)
The effect of improving the susceptibility to infection in the cellular immunosuppressive state during stress loading was evaluated.
As a test sample, muirapuama extract or dicoppi was added to a 0.2% CMC solution at a rate of 20,1000 mg / 10 ml, respectively, and stirred to make it uniform. In addition, muirapuama extract and dicoppi were added to a 0.2% CMC solution at a ratio of 20,1000 mg / 10 ml, respectively, and stirred to make uniform.
As a control sample, a 0.2% CMC solution used for crude drug suspension was prepared.
Seven-week-old male C3B6F1 mice were used as test animals. The above samples were orally administered once a day for 7 days every day from 5 days before stress loading by a syringe for oral administration at a dose of 0.25 ml / head.

Each test sample was stressed using a mouse restraint cage for 14 hours at night (18:30 to 8:30 the following), which is the activity time of the mice for 3 days from the 5th day of administration. Immediately after the end of stress loading on the first day, Listeria monocytogenes (5.6 × 10 ⁵ CFU / head) was intraperitoneally inoculated, and the number of Listeria monocytogenes infections in the liver on the third day after infection was measured. The period of stress load was fasting and fasting. The control sample administration group was similarly tested.

The results are shown in FIG. In statistical processing, the average value of the comparison target was tested by Student t-test, and p <0.05 was considered significant.
There was no change in the number of Listeria monocytogenes infections in the group administered with muirapuama and dicoppi alone compared with the control sample administration group. On the other hand, the combined group of muirapuama and dicoppi showed a significant decrease in the number of infectious bacteria compared to the control sample administration group.

  Therefore, a combination of muirapuama, which has the effect of appropriately maintaining the blood glucocorticoid concentration that rises when stress is applied, and dicoppi, which has the effect of promoting the production of interferon γ (hereinafter abbreviated as “INFγ”) from spleen cells in the presence of glucocorticoid. Has been found to have an effect.

It is the figure which showed the time-dependent change of the blood corticosterone density | concentration by stress load, the vertical axis | shaft showed the blood corticosterone density | concentration and the horizontal axis | shaft showed the time from the stress load start. * Indicates a significant difference between the stress group and the non-stress group. † indicates a significant difference between the stress group and the muirapuama administration group. A graph showing the amount of each IFN-γ produced when the test sample is added, with the control sample IFN-γ production from spleen lymphocytes stimulated by ConA in the presence of corticosterone (50 nM) as 100%. is there. It is the figure which showed the number of infection in the liver 3 days after Listeria monocytogenes infection. * Indicates a significant difference between the stress group and the non-stress group. ** indicates a significant difference between the RU486-inoculated group and the stress group. It is the figure which showed the number of infectious bacteria in a liver 3 days after Listeria monocytogenes infection of a test sample administration group and a control sample administration group. * Indicates a significant difference in the combination of muirapuama and dicoppi with respect to stress.

Claims (6)

  A composition comprising (a) muirapuama and (b) one or more selected from kojishiki, jikoppi, halibut, zion, ginkgo beige, keisan leo, tennansho and iodope.   A composition comprising muirapuama and dicoppi.   The composition according to claim 1, which is an antistress agent.   The composition according to claim 1, which is an agent for improving stress susceptibility.   The composition according to claim 1, which is an agent for preventing or treating infectious diseases. The composition according to claim 5, wherein the infection is common cold or influenza.

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