JPS6245526A - Agent for ameliorating functional disorder caused by stress - Google Patents

Abstract

PURPOSE:To provide an agent for ameliorating functional disorder caused by mental stress, containing at least one kind of a phenylpropanoid glycoside as an active component, having anti-stress effect and low toxicity and especially effective to ameliorate and cure the hypogonadism and amnesia, etc. CONSTITUTION:The agent contains the phenylpropanoid glycoside of formula (R<1> and R<2> are H, OH or methoxy; R<3> is H, beta-D-glucopyranosyl or beta-D- apiofuranosyl; R<4> is H or acetyl) as an active component. Especially preferable examples of the compound are acteoside (R<1> and R<2> are OH; R<3> and R<4> are H), echinacoside (R<1> and R<2> are OH; R<3> is beta-D-glucopyranosyl; R<4> is H), etc. The functional disorder caused by stress, especially hypogonadism, amnesia and psychosomatic disease can be cured by the agent. Dose: 0.05-5g daily by oral administration.

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial Application Field) The present invention relates to an agent for improving stress-induced dysfunction, and particularly contains phenylpropanoid glycosides as an active ingredient.

Pertains to a stress-induced functional impairment improving agent.

(Prior art) Phenylpropanoid glycosides have the formula: R4 means a sil group, and R4 means a hydrogen atom or an acetyl group). Conventionally known phenylpropanoid glycosides include acteoside (R1, R2: hydroxy Examples include the following compounds, including groups R3, R4: hydrogen atoms.

Ethinacoside (R', R12: hydroxy group, R3: β-D-glucopyranosyl group, R4, hydrogen atom), 2'-acetyl acteoside (R1, R2: hydroxy group, R1: hydrogen atom).

R4: acetyl group), ottomansuside B (R1, R2, R3, R4: hydrogen atom), forcisoside B (R'', R2: hydroxy group, R3: β-D-apiofuranosyl group, R4: hydrogen Regarding the pharmacological and physiological effects of these phenylpropanoid glycosides, it has been reported that both echinacoside and forcisoside B have antibacterial effects (A,
5tal1 etc.11 Helv, Chim.

Acta,” p. 238, 1950 and Hayato et al., “Proceedings of the 28th Annual Meeting of the Japanese Society of Herbal Pharmaceutical Sciences,” p. 20, 1981), and acteoside has antiparkinsonism and β
-It has been reported that it has a blocking effect [West German Patent Application Publication No. 2609533] and that it has an antitumor effect (Numata et al. 33rd Kinki Branch of the Pharmaceutical Society of Japan) General Meeting Lecture Abstracts 2017, page 76, 1983
Year).

On the other hand, although the parasitic plant of the Physcomitraceae family is listed in the oldest herbal book, the Shennong Bencaojing, and has been used as a tonic since ancient times, there is no pharmacology to support its tonic effect. The reality is that its physiological effects have not been proven, and only a slight effect of lowering blood pressure and promoting saliva secretion have been reported. (2013) and ``Summary of Obsolete Research Literature,'' p. 259, 1975). For this reason, the present inventors have discovered the traditional efficacy of meat
We understand that this is synonymous with the healing of various symptoms caused by stress-induced decline in the function of biological organs and mental fatigue, and as a result of repeated research on Nikkujiyo focusing on this point, we found that Nikkujiyo has In addition to the various known phenylpropanoid glycosides, he discovered that the following new phenylpropanoid glycosides were contained, and reported this and filed a patent application for the isolation method (Kobayashi) etc. rchem, Phar
m, Bul, 1. J Volume 32, No. 3009 and 388
0 page, 1984 and Japanese Patent Application No. 60-29335).

Cystanoside A (R1: hydroxy group, R2: methoxyl group, R3
: β-D-glucopyranosyl group, R4: hydrogen atom) Cystanoside B (R1, R2: methoxyl group, R3: β-D-glucopyranosyl group, R4: hydrogen atom) Cystanoside C (R1: hydroxy group, R2: methoxyl group, R3,
R': hydrogen atom) and cystanoside D (R1, R'': methoxyl group, R3, R4, hydrogen atom) As a result of preliminary tests, these cystanosides were estimated to have anti-stress effects. has not been fully elucidated.

Furthermore, the animal testing method for proving the medicinal efficacy of crude drugs, which are classified as supplementary medicines in Oriental medicine, was established by Saigo et al.
66, 1983), Eleuthero extract (Japanese Unexamined Patent Publication No. 59-534) is effective in recovering functional disorders caused by stress.
26), Syringin (JP 59-33221), Syringaresinol diglucoside (JP 59-1162)
20) and iridoids, glycosides or salts thereof (JP-A-59
-164717) has been reported to be acceptable.

(Problems that inventions aim to solve) In modern times, where scientific and technological innovations are progressing rapidly, social mechanisms are becoming more complex, and as a result, people are forced to live lives that are subject to a lot of stress. As a result, psychosomatic disorders and neuroses tend to occur frequently. Psychosomatic disorders appear as disorders in the respiratory system, digestive system, circulatory system, urogenital system, etc., and neurosis appears as insomnia, emotional instability, amnesia, etc. Therefore, in order to lead a healthy social life in the modern living environment, pharmaceuticals that improve physical and mental dysfunction caused by stress are extremely important.

Therefore, the problem to be solved by the present invention is to provide this type of medicine, especially an anti-stress agent containing substances obtained from herbal medicine as an active ingredient, thereby improving functional disorders caused by stress. .

(Means and effects for solving the problem) As mentioned above, it has been reported that phenylpropanoid glycosides have a few pharmacological and physiological effects, but anti-stress effects was previously unknown.

The inventors of the present invention have used five-labor-seven-skin, yang-wi, and
It is a herbal medicine that is said to be effective for amnesia, cold pain in the lower back, enuresis, infertility, leucorrhoea, constipation, etc., and can be used alone or with other herbal medicines, as well as in Chinese herbal prescriptions such as Nikshuyogan, Kanshotan, and Shuyojunchogan. We have also been conducting research on niku kongo used in medicinal alcoholic beverages, and as a result, we have announced that niku junin contains various phenylpropanoid glycosides, as mentioned above. but,
After that, further research was carried out, and the pharmacological and physiological effects of both parts of each stage obtained during extraction of Nikkongyo and the constituents obtained by further purification of these parts were investigated in the animal tests described by Soran et al. As a result of investigation in accordance with the law, it was found that stress-induced sexual behavior disorder and decline in learning and memory ability can be improved by administering phenylpropanoid glycosides, leading to the completion of the present invention.

Therefore, according to the present invention, it contains as an active ingredient at least one phenylpropanoid glycoside represented by the formula (wherein R", R", R' and R1 have the meanings given in 1)q. It is characterized by
An agent for improving functional impairment caused by stress is provided.

The active ingredients of the agent according to the present invention are: I'-Chick purified acteoside, echinacoside, cistanoside A
Alternatively, cystanoside C or the like is advantageous, but it can be used even if it has not been isolated and purified in this way.

(Dosage form and dosage) There are no particular restrictions on the dosage form of the agent according to the present invention, and therefore, at least one compound of tephenylpropanoid glycosides may be administered as such, or it may be administered after being formulated. When formulated, it can be made into oral preparations such as powders, fine granules, granules, capsules, liquids, syrups, etc., or parenteral preparations such as injections and intestinal injections. You can also do that.

The dosage depends on the patient's age, weight, symptoms, dosage form, etc., but when administered orally to adults, the dose is 0.05-5 phenylpropanoid glycosides divided into 1-3 doses per day. It is appropriate to take and use it.

(Effects of the Invention) The present invention is based on the discovery and confirmation of new pharmacological or physiological effects and anti-stress effects regarding phenylpropanoid glycosides. Disorders of various functions, especially sexual dysfunction and amnesia, can be alleviated or cured, and psychosomatic disorders can be prevented and treated.

Furthermore, the toxicity of phenylpropanoid glycosides is extremely low;
Therefore, the agent according to the present invention has excellent safety in use.

(Formulation Examples, etc.) Next, the present invention will be explained in more detail with reference to production examples, pharmacological and pharmacological test examples, and formulation examples of phenylpropanoid glycosides.

yλ = 迭banoid drunken prisoner! Area 11: Phenylpropanoid glycosides are found in lilac, osmanthus, osmanthus, privet, etc. in the Oleaceae family, in lilacs in the Oleaceae family, in Nicotiana tabacum, etc. in the Nicotianaceae family, in Physcomitrella sinensis, Nikkisei, etc. in the Mouseaceae family, and in Lamiaceae lilac, osmanthus, and other plants in the Lamiaceae family. , and is also contained in trumpet, etc. of the family Phosophyllaceae, and therefore, it can be obtained by extracting these plants as raw materials. Considering the fact that there are many types of phenylpropanoid glycosides contained among these raw materials, meat ゛4゜蓜蓝 was selected as a raw material, and this was used in Kobayashi et al.'s rChem.

Pharm, Bull, J Vol. 32, pp. 3009 and 3880, 1984 and patent application 1984-2933.
A phenylpropanoid glycoside was prepared by processing according to the method described in No. 5 specification.

That is, the meatballs are cut into small pieces, extracted with an aqueous alcohol solution (water and a highly polar organic solvent can also be used together), and the resulting extract or its n-butanol soluble portion is added to a styrene-based non-polar adsorption resin, such as Commercially available Diaion IIP-20 (
manufactured by Mitsubishi Chemical Industries, Ltd.) and Amberlight XAD-2
(manufactured by Rohm and Haas) to obtain a crude glycoside fraction (hereinafter referred to as "both fraction-1"). Next, these two parts-1 are mixed with a polyamide resin such as commercially available polyamide C-2.
00 (manufactured by Wako Pure Chemical Industries, Ltd.) to form a mixed fraction of crude iridoid glycosides and crude lignans (hereinafter referred to as "fraction-2") and crude phenyl glycosides. Propanoid glycoside fraction (hereinafter referred to as "Fraction-3")
It was separated into two parts.

The results of thin layer chromatography analysis of both fractions-2 under the following conditions are shown in Figure 2.As is clear from the analysis results, both fractions contain 8-ebroganic acid (1), Ajugol (2), Musaenosidic acid (
3), genipocidin M (4), 6-dioxycatalbol (6), glucoside (7).

Iridoids and their glycosides such as baltosioside (8), 8-epideoxyloganic acid (9), cystanine (12), syringaresinol diglucoside (5),
Noridanane glycosides such as syringaresinol monoglucoside (11) and 8-hydroxygeraniol-1-
Compounds such as glucoside (10) are included, but
Contains no phenylpropanoid glycosides.

Thin plate Nijirikageru 60F2. , (manufactured by Merck) Developing solution: Chloroform/methanol/formic acid/water = 70
/ 30 / 2 / ] Color development method = 20ν/v (1) After spraying sulfuric acid, heating at 105°C. On the other hand, thin layer chromatography analysis was performed on both parts -3 under the following conditions. The results are shown in Figure 1. As is clear from the results of this analysis, both of these components contain cistanosides A-D (2.4, 5, and 8 in the figure), which are phenylpropanoid glycosides.

Ethinacoside (1), acteoside (3), 2'-
Acetyl acteoside (6) and ottomanside B
(7), but no iridoid or lignan-related compounds.

Thin plate Nijirikageru 60F2. (Manufactured by Merck) Developing solution: Chloroform/methanol/water = 6/4/1 Color development method: 20 v/v (%) 105 after spraying with sulfuric acid
Various phenylpropasoyl contained in 1-3 heated at ℃
~To obtain 9 lIi of each glycoside, one column of 1-3 was subjected to silica gel column chromatography and non-polar adsorption resin (
For example, it may be treated with 1 gram of Sephadex L11-20) column chroma manufactured by Pharmacia Fine Chemicals.

1 effect, 1 night, 1 night boiling - Soran et al.'s method (``The 16th Japanese and Chinese Medicine Symposium Abstracts,'' p. 66, 1983) is used to treat sexual behavior disorders and learning and memory disorders that occur after stress loading. The improvement effect of phenylpropanoid glycosides was investigated.

(1) Experimental Animals Select RV-CS H male mice (weight 28-32 g) that are 9 weeks old or older and have normal sexual behavior, and
The animals were reared under conditions of ±1°C and humidity of 55±5%.

In this case, "normal sexual behavior" means that male mice raised in private cages are allowed to cohabit with 10 female mice that have been brought into heat by subcutaneously administering 10 μg/kg of estradiol every day for 105) days. Point out when you successfully complete the intro mission 5 or more times in a week.

(2) Stress loading A male mouse, which is an experimental animal, was suspended in the air and fixed at a height such that the tip of its nose touched the water surface.

This state was maintained for 30 minutes on the first day of the test, the suspended time was gradually extended from the second day onwards, and on the final day of the test, the 15th, the subjects were maintained in the suspended state for 2 hours to apply stress. Even when continuous stress was applied for 15 days using this method, no weight loss was observed compared to the control non-stressed group, and motor coordination disorders, decreased muscle tone, and decreased spontaneous and exploratory movements were observed. However, a decrease in sexual behavior and learning and memory behavior was observed.

(3) Experimental method and method for evaluating the effect of test drug The stress load was carried out every day from the specified time (1:00 p.m.), and the test drug was orally administered after the stress load.

The next morning, sexual behavior will be examined from 9:00 am and learning (memory) behavior will be examined from 11:00 am, and this will be repeated for 15 days to examine the improving effects of the test drug.

a) Determination of sexual behavior Estradiol was administered daily (10 gg/kg)
Cages containing 10 female mice each (30 x 40
One male experimental animal mouse was allowed to live with the mouse in a 20 cm x 20 cm room for 10 minutes, and the number of times each sexual behavior of licking, mounting, and intromission was performed during that time.
Measure the number of actions and the time it takes to take action. In this case, there were 10 animals per group, and the χ2 test was used for the number of significant differences between groups, and the analysis of variance method was used for other cases.

b) Judgment of learning (memory) behavior A rubber stopper (diameter 5 cm, height 5 am) was placed on the floor where a current (DC 36 V, O, 1 mA) was flowing, and a male mouse, which was an experimental animal, was Let them learn not to get off the rubber stopper for a few minutes. A male mouse subjected to this learning procedure was placed on a rubber stopper, and the number of times it accidentally got off the rubber stopper within 5 minutes and the number of times it got off incorrectly out of 110 mice were calculated as dI. Established. In this case, to test for significant differences between groups, analysis of variance was used for the number of incorrect landings, and χ2 test was used for the four reasons for incorrect landings.

(4) Results and analysis 8) Effect on sexual behavior j) Results The results of investigating the decrease in sexual behavior due to stress load and its recovery due to administration of each test drug were as shown in the table below.

ii) Analysis The sexual behavior of male mice follows the steps of linking, mounting, introduction, and ejaculation.

Therefore, the effectiveness of the treatment can be determined by examining what changes occur in the sexual behavior of male mice treated with female mice in heat. In other words, it is determined that the more often a group of 10 animals performs each sexual behavior within a certain period of time, the more times they engage in sexual behavior, and the shorter the time it takes to initiate sexual behavior, the more active their sexual behavior is. I can do it.

As is clear from the above table, the sexual behavior of male mice subjected to stress is generally reduced. When comparing the stressed test group and the non-stressed control group, it became clear that the difference was more pronounced in later stages of sexual behavior than in licking, which is the early stage of sexual behavior. . Regarding the number of times an individual mouse engages in sexual behavior and the time required to initiate it, stress load has relatively little effect on the number of times an individual mouse engages in sexual behavior, but the number of times an individual mouse engages in sexual behavior A clear difference was observed in the time required to start the process.

Thus, by administering the test drug to male mice whose sexual behavior had decreased due to stress, a clear improvement in sexual behavior was observed. Regarding the relationship between the type of test drug and its improving effect, Ryoubun-1 (crude glycoside fraction, which also contains various phenylpropanoid glycosides and lignan and iridoid-related compounds) was found to be effective. The degree of improvement was the lowest, and the improvement effect was observed at a dose of 50 mg/kg. The latter, that is, phenylpropanoid glycosides, was found to be more effective when compared with those containing glycosides, and actiosites obtained by further purifying fraction-3, It was found that ethinacoside, cistanoside A, and cistanoside C can produce effective improvement effects with a relatively small dose of 10 mg/kg.

These facts indicate that phenylpropanoid glycosides have the effect of effectively improving the decline in sexual function caused by stress.

b) Effect on learning (memory) behavior i) Results The test results were as shown in FIG.

ii) Analysis: In the non-stressed control group, the number of times they accidentally fell off the rubber stopper during the 5-minute test period was an average of 0.75 times per day during the 15-day test period. In the stressed test group, this was an average of 2.6 times a day, indicating that stress caused a decline in learning behavior.

By administering the test drug to male mice whose learning behavior had decreased due to stress, the number of times they accidentally fell off the rubber stopper was clearly reduced. A similar trend was also observed for the number of mice that made errors.

Furthermore, the action of echinacoside is weaker than that of other phenylpropanoid glycosides, and the action of Ryobu-2, which consists of lignans and iridoid-related compounds, is similar to that of various phenylpropanoid glycosides. Ryobu as a component -3
The effect was weaker than that of individual phenylpropanoid glycosides other than ethinacoside.

These facts indicate that phenylpropanoid glycosides have the effect of improving the decline in memory ability caused by stress.

(Toxicity test) ddY mice (weight 25-30g) were kept at a temperature of 23±1°C.
.

Healthy individuals were preliminarily reared for one week under conditions of ad libitum feeding and water supply in a room with a humidity of 55±5%, and a group of five animals was selected. Then, after fasting for 18 hours, they were given 1 g/kg each for echinacoside, acteoside, and cystanoside A, and 500 mg/kg for cystanoside C by three routes: oral, intraperitoneal, and intravenous. administered.

For all test drugs and for all administration routes, sedation and hypothermia were observed for approximately 5 hours immediately after administration, but recovery occurred approximately 6 hours later, and no deaths were observed. After that, body temperature and general symptoms were examined for one week, but no differences were observed between the control group and the control group.
No abnormalities were found upon autopsy.

This shows that the toxicity of phenylpropanoid glycosides is extremely low, suggesting that it is extremely safe when used as an active ingredient in pharmaceuticals.

Formulation Example 1 (Granules) A crude extract obtained by immersing Physcomitrella sinensis in a 50% ethanol aqueous solution was dissolved in water and subjected to Diaion 1 (P-20 column chromatography treatment to obtain the crude glycoside fraction (fraction). -1) was obtained. Both fractions were subjected to polyamide column chromatography to obtain a crude phenylpropanoid glycoside fraction (fraction-3).

95g of starch and 980g of milk were added to 20g of both, mixed uniformly, and then hydroxypropylcellulose (
5g) Wet granulation using an ethanol solution in a conventional manner,
It was dried and sized to obtain granules.

1 stable! (Capsules) 100 g of lactose and 85 g of starch were added to Echinacoside Log and mixed uniformly, then wet granulated using an ethanol solution of hydroxypropyl cellulose (5 g) by a conventional method, dried, and sized. Hard capsules were obtained by filling 200 mg per capsule.

Juranko" (tablets) 90 g of starch, 97 g of lactose, hydroxypropyl cellulose 'Ig and 2 g of magnesium stearate were added to 10 g of acteoside, mixed uniformly, wet-granulated by a conventional method, dried, and sized, The mixture was compressed to obtain seven 200 mg tablets.

(Liquid for internal use) Cystanoside A iog, 100g of glucose,
An appropriate amount of fragrance was dissolved in a 14% ethanol solution to give a total volume of 10,100O to obtain a liquid preparation for internal use.

[Brief explanation of the drawing]

Figure 1 is a thin-layer chromatogram of crude phenylpropanoid glycosides (fraction-3) extracted from Mekonggyong;
The figure is a thin-layer chromatogram of iridoid and lignan-related compounds (fraction-2) extracted from Physcomitrella sinensis.
The figure is a graph showing the effects of stress on the learning and memory behavior of mice, and the ameliorating effects of Ryoubun-2, Ryobun-3 and each phenylpropanoid glycoside on the decline in learning and memory behavior. Figure 1 *: **; Strike 4! Negative Kamitsuji sheep stock suction fraction 3ff-5# (20mg/Kg 7 days) [)<0.0
5pku0. O1

Claims (5)

[Claims] (1) Formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (In the formula, R^1 and R^2 mean a hydrogen atom, a hydroxyl group, or a methoxyl group, and R^3 is a hydrogen atom, β-D-
Glucopyranosyl group or β-D-apiofuranosyl group, R^4 means hydrogen atom or acetyl group) Contains at least one phenylpropanoid glycoside as an active ingredient. characterized by
An agent for improving functional disorders caused by stress. (2) R^1 and R^2 mean a hydroxy group, and R
The functional disorder improving agent according to claim 1, characterized in that the active ingredient is ethinacoside, in which ^3 means a β-D-glucopyranosyl group and R^4 means a hydrogen atom. (3) R^1 and R^2 mean a hydroxy group, and R
The functional disorder improving agent according to claim 1, characterized in that the active ingredient is acteoside in which ^3 and R^4 represent hydrogen atoms. (4) Effective cistanoside A where R^1 means a hydroxy group, R^2 means a methoxyl group, R^3 means a β-D-glucopyranosyl group, and R^4 means a hydrogen atom. The functional disorder improving agent according to claim 1, characterized in that it is a component. (5) A patent characterized in that the active ingredient is cistanoside C, in which R^1 means a hydroxy group, R^2 means a methoxyl group, and R^3 and R^4 mean a hydrogen atom. The functional disorder improving agent according to claim 1.