JPH0459733A - Functional disorder improver caused by stress comprising extracted and purified substance of cnidii monnieri fructus as active ingredient - Google Patents

Abstract

PURPOSE:To obtain the title improver capable of alleviating and curing disorders of various functions caused by stresses, comprising an extract having high polarity extracted from Cnidii monnieri Fructus or a benzofuran contained in the extract as an active ingredient. CONSTITUTION:At least one selected from an extract having high polarity obtained by extracting fruit of Cnidii monnieri Fructus with an extraction solvent comprising water or a hydrous organic solvent and removing the extraction solvent, or a compound shown by the formula (R<1> is H or methyl; R<2> is H or beta-D-glucopyranosyl) contained in the extract and a glycoside thereof as an active ingredient. The compound shown by the formula is obtained by dissolving the extract in water, subjecting to column chromatography using a porous styrene-based resin and to column chromatography using a filler such as higher alkylsilanol resin, Sephadex or silica gel. The improver is useful as vitalizing agent, tonic and psychomimetic agent.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of the Invention The present invention relates to an agent for improving stress-induced dysfunction, which contains as an active ingredient a highly polar extract extracted from Jasperum or benzofurans contained in the extract. .

Conventional technology Snake beetroot has been used since ancient times, as it is listed in the oldest Chinese herbal book, Shennong Cao Ching, under the name ``Jashoushi''; it is one of the herbal medicines. There is, and in Chinese medicine prescriptions, Sankomaru (Senkinpo: Jatoko, Mishiko,
Schisandra), Jatoko-to (medical sect Kinpo: Jatoko, Eireisen, Tokio, Oga, bitter ginseng, sand grain, green onion head), and single ingredients include Jatoko Yosan (Kanaki main route: Jatoko) Even today, it is considered a valuable material.

Cnidii monnleri Fruci
us) is a member of the family Apiaceae (Umbelliferae).
, (=Selium monnieri L,)) is dried. This okazeri is a biennial plant that grows wild in northeastern China, especially in Kahoku, wild plants, abscessive,
Produced in Jiangsen and Siyong provinces, the stems are 40-70cm tall.
The stem is hollow and has longitudinal grooves and knots. The leaves are stalked and twice pinnately lobed, and the lobes are oblong and 2 to IOC++ long! , radial ~3
cm, pinnate, flowers are white, fruits are small oval and 2 long.
~3 cm, 1-2 CTn double hanging flowers, glabrous and pale brown, but its congener plant H6racleumsc
abridum Franch, Tori
lis japonica (Houtt, )), 5cabra (Houtt, no), and 5cabra (Houtt, no) are also considered to be the progenitor plants of Jatoko ('Tsuneo Mami: “Illustrated Encyclopedia of Primary Colors of Japanese and Chinese Medicine (Part 1)” p. 233-234 (1980), Kinyoshi Akamatsu: Newly revised Japanese and Chinese Medicine p., 188 (1970)).Among them, the Japanese herb louse is sometimes used for the same purpose as a Japanese snake bed.

Next, Jaboko contains 1.3% of essential oil, the main components of which are Q-bin, camphene, isovaleric acid bornyl ester, and the presence of various coumarins such as osthole. It has also been reported (Tsuneo Namba: “Illustrated book of primary colors of Japanese and Chinese medicine (Part 1)” p. 234 (1980), Tatsuo Karimae:
Latest Pharmacognosy p, 395 (1987), Xiang:
Zhongguo Yaoke Daxue Xue
bao 20(2) p, 92-93 (1989)).

On the other hand, its pharmacological and physiological activities include antifungal components such as essential oils (B. epidermis purpura, gypsum-like microspores,
Antiviral, anthelmintic, and antitrichomoniacal effects have been reported (12i Wave Tsuneo: “Primary Colored Japanese and Chinese Medicine”).
Above) p-234 (1980), Oiu, 5hiyu:
CN 85103316). In addition, anti-mutagenicity (Liu, D: Bull chin mate)
r med 13(11) (1988)), antiallergic effect (Joji Yamahara: Chem, pharm, b
ull, 33(4) (1988)), antitumor effect (
Supervised by Shoji Shibata: “Drug Development Volume 2 - Pharmacologically Active Substances n
"p, 416 (1989)), local anesthetic effect (L
ianO:Bull chin mater med
13(9) p. 40-42 (1988)), as well as
Recently, the calcium antagonistic effect of ostole in the ingredient (Joji Yamahara: Yakushu 105 (5) p. 449-458 (198
5)), and it is expected to be used as an antihypertensive agent. As described above, regarding the components and pharmacological/physiological activities of the herbal medicine, essential oils (Q-pinene, camphene, bornyl isovalerate), coumarins (colum.
bianadin, O-acetylcolumbi
anetinzcolumbianetin, ○-1s
ovaleryl-columbianeLin.

cnidiadin, arch-angellcin
Sedultin, 2'(S), 3'(R)-isobutyryl oxine-〇-acetyl-2', 3'-dihydrooroserol, bergapten, isobimbifurin, oroseron, ostol, etc.), a- candinenelltorilli
Research has mainly been carried out on fat-soluble components such as n, and there are still many unknowns about the other components and their pharmacological and physiological activities, and these have not yet been elucidated.

In particular, there have been no reports regarding benzofuran derivatives or their glycosides as constituents of the herbal medicine Jatoko. In addition, regarding the pharmacological and physiological activities of these substances having a benzofuran skeleton, for example, amiodarin
arine) antiarrhythmia effect (TheMerck In
dex 11 edition p, 501 (198
9)), Antifungal action of hordatin (hordat 1ne) (supervised by H, Wagner and P, Wolff:
“New natural products and
plantdrugs with pharmaco
logical, biological orthe
rapeutical activity''p, 227
(1977)) are only known.

Problems to be Solved by the Invention As mentioned above, what has been elucidated so far regarding the ingredients, pharmacology, and physiological activities related to Jabedoko is limited to fat-soluble ingredients such as essential oils and coumarins. In terms of its effects, it is different from the traditional efficacy of Jabedoko, such as antifungal, local anesthetic, antiallergic, antitumor, and calcium antagonistic effects. .

In other words, the traditional efficacy of this jaboko is interpreted as relieving and curing various symptoms caused by the decline in biological functions caused by various types of stress and mental and physical fatigue.

A wide variety of stress factors can be considered as characteristics of modern society. For example, due to advances in electronics, companies'
In addition to various stresses related to social problems such as techno stress caused by the aging society and the rapidly increasing number of elderly people in an aging society, there is also stress associated with the speeding up of information in the international information age, and the advancement of women in the workforce. We live in an era of excessive stress, as it is said to be the cause of various types of stress. For this reason, there is a strong demand in modern society for the development of safe anti-stress drugs.

Under these circumstances, the present invention was made for the purpose of providing the above-mentioned medicines from a snail.

Means for Solving the Problems In addition to the above-mentioned traditional efficacy of Jabedoko, the present inventors have conducted intensive research focusing on the relatively highly polar component, which has not been studied much in the past. It was confirmed that the highly polar extract has a safe anti-stress medicinal effect. Furthermore, this crude extract contains Columbianadin, O-acet, which is a compound already known from Jatonicum.
ylcolumbianetin.

Columbianition, Oolsova 1
ery Ico lumb 1ane tin.

Cn1diadin, Archangellcin,
Edultin, 2'(S), 3'(R)-isobutyryloxy-〇-acetyl-2', 3'-dihydrooroserol, bergapten, isovinpilin, oroseron, ostol,
In addition to a-candinene and torillin, 8-hydroxy 3(S)-linalool 3-0-β-
In addition to clarifying the presence of monoterpenoid compounds such as D-glucopyranoside and 6-hydroxylinaroyl oxide, we also found that new benzofurans represented by the following general formula (1) are contained, and that the benzofurans have the above-mentioned medicinal properties. We found that this shows that

As described above, the inventors have found that the above-mentioned serpentine extract and purified products (benzofurans) are effective as an ameliorating agent for stress-induced functional disorders, and have completed the present invention based on this knowledge.

That is, the present invention provides a highly polar extract obtained by extracting the fruit of the serpentine fruit with an extraction solvent consisting of water or a water-containing organic solvent, and then removing the extraction solvent, or - A stress-resistant product characterized by containing as an active ingredient at least one selected from benzofuran derivatives and glycosides thereof represented by a hydrogen atom or a methyl group, and R2 represents a hydrogen atom or a β-D-glucopyranosyl group. The present invention provides a functional disorder improving agent.

These active ingredients are preferably isolated and purified, but
It can also be used in the form of both components containing at least one of these purified products.

The new benzofurans represented by formula (I) are as follows:
It is a species compound.

(1) Kunideoside A (R1”' H, R1!β-D
・Glucopyranosyl group) 4-oxy-4,5-floromelilorinoc annod 2
-0-β-glucopyranoside (2) Kunideoside B (R, = metquin group, R2 = β
-D-glucopyranosyl group) 4-oxy-4,5-flo 3-methquine meliloric acid 2-0-β-glucopyranondo (3) Cnideol b (R, = methoxy group, R2 = H) 4-oxy-4 , 5-Flow 3-Methoxymeryloric acid The benzofuran derivative of the formula (I) can be obtained by subjecting the benzofuran derivative of the formula (I) to solvent extraction of serpentium oleracea, dissolving the obtained extract in water, and subjecting it to column chromatography using a porous styrene resin. After applying it to
Repeatedly subjecting the obtained crude benzofuran derivative mixed metal compound to column chromatography using at least one filler selected from higher alkylsilanol resins, Sephadex, silica gel, celite, and porous styrene resins. obtained by.

In this method, water, organic solvents, and water-containing organic solvents can be used as extraction solvents. Preferred organic solvents are highly polar ones, such as methanol, ethanol, acetone, dioxane, dimethyl sulfoxide, and the like. As a water-containing organic solvent, the organic solvent concentration is 20 to 70%.
(v/v), particularly preferably 40 to 60%. The raw material Jabedoko can be cut into small pieces and then extracted, or can be defatted according to a conventional method and then subjected to extraction. It is preferable to use the extraction solvent in an amount 4 to 10 times the amount of the crude drug.

The extraction time varies depending on the type of extraction solvent and temperature conditions, but if the extraction process is carried out at room temperature using 5 times the amount of water-containing methanol on the shredded scallops, the extraction time will be approximately 2 to 4 hours. It is appropriate to repeat 2rff1 times. After each extraction is completed, the extracts obtained by filtration are combined in a conventional manner and the extraction solvent is removed to obtain a crude extract. As a method for removing the extraction solvent, a conventional concentration method such as a vacuum concentration method or a spray drying method can be used. Since the obtained crude extract contains fat and the like, if necessary, the extract is suspended in water and treated with a low polar organic solvent to remove fat. Examples of the low polar organic solvent include hexane, benzene, ether, and ethyl acetate. However, usually the crude extract is suspended in water, and the aqueous solution from which water-insoluble parts are removed is directly treated with a porous styrene resin. As a specific commercial product of this porous styrene resin, for example, styrene-based Diaion HP
/ Leeds (manufactured by Mitsubishi Chemical Industries, Ltd.), polystyrene-based MC
Examples include lgel CHP 20P (manufactured by Mitsubishi Chemical Industries, Ltd.). In this treatment, desired benzofurans and the like are adsorbed onto the resin by passing the above aqueous solution through a column filled with an adsorption resin. Next, after washing with a predetermined amount of water, benzofurans and the like adsorbed on the resin can be eluted using a highly polar organic solvent or a water-containing highly polar organic solvent.

As this highly polar organic solvent, for example, methanol, ethanol, acetone, dioxane, dimethyl sulfoxide, etc. can be used, but especially when the methanol concentration is 4.
0 to 60% (V/V) aqueous methanol is preferred.

Among the benzofurans, those in which R2 in formula (I) is a glucopyranosyl group are preferably used in higher alkylranol-based resins such as octate'yl/lanol (hereinafter abbreviated as ○DS) and Sephadex-based resins. After removing the eluate, the treatment is repeated in a column filled with these resins, and the liquid is separated and purified by sequentially changing water, water-containing organic solvent, and organic solvent, resulting in a pale yellow color. A powder of Cnideoside A1 Cnideoside B is obtained. On the other hand, among the benzofurans of the present invention, those in which R2° in formula (1) is a hydrogen atom are preferably adsorbed on silica gel or oDS-based resin. By treating with a column filled with these resins and repeating solvent treatment,
Cnideol b can be obtained as a pale yellow powder. In addition, during these purification operations, a new monoterpene derivative, 8-8-
It is also possible to obtain 6-hydroxylinaroyl oxide from hydroxy-3-(S)-linalool and from both of the more polar components.

Dosage Form and Dosage The dosage form of the agent for improving stress-induced dysfunction of the present invention is not particularly limited. Both components can be administered as they are, or they can be formulated and administered. For formulation, powders, fine granules, granules, tablets, capsules, tea preparations, liquid preparations (alcoholic preparations,
Forms for internal use such as tinctures, liquid extracts, syrups, etc.) are preferred. Note that solid or liquid excipients used here are those known in the art. Namely, to list some of them, there are lactose, starch, dextrin, calcium phosphate, synthetic and natural aluminum cinnamate, magnesium oxide, dried aluminum hydroxide, magnesium stearate, dried yeast, etc.
Excipients used in liquid preparations include water, glycerin, monosilox, propylene glycol, ethanol, fatty oil, ethylene glycol, polyethylene glycol, and sorbitol.

The dosage of the present agent varies depending on the type of active ingredient, dosage form, patient's age, weight, disease process, etc., but the crude extract is usually 1 to 100 g, preferably 1 to 100 g, as lFE amount for adults.
10 g, especially 3 to 9 g, of purified products (Fraction ①: mixture of crude Kuri deoside A and B, fraction B: high purity Kuri deoside A
, B mixture, fraction V: crude chlorideol), usually 0.0
05 to 19, preferably C 0.05 to 0.6 g, especially fractions m and V tesoreso hO, 3 to 0.6 g, fraction B 0.1
It is appropriate to take ~0.3g orally in 3 to 4 divided doses per day.

Effects of the Invention The agent of the present invention has an anti-stress effect and can reduce and cure various functional disorders caused by stress, such as sexual dysfunction, and also quickly recover from mental and physical fatigue. It is suitable for use in the prevention and treatment of psychosomatic disorders caused by these diseases, and is therefore useful as a tonic, a tonic, a psychoactive agent, and the like.

In addition, in acute toxicity tests using mice, the crude extract and purified product used in the present invention showed that even though the former had a concentration of 109/kg and the latter had a concentration of 39/kg, within 72 hours after oral administration, there were no cases of death or noticeable death in the applied animals. No change was observed, and therefore, the agent of the present invention is extremely safe to use.

Example Next, the present invention will be explained in more detail with reference to Examples.

Production example: 50% for 1.5 kg of shredded snake roe (made in China)
Extraction treatment with (v/v) methanol aqueous solution 8Q at room temperature for 3 hours was repeated twice. These extracts were combined and concentrated under reduced pressure to obtain 139.09 g of crude extract.

This was suspended in water 1501RQ, and the aqueous solution (crude extract content: 116.h) from which water-insoluble parts were removed was passed through a column packed with polystyrene resin MCI get CHP 20P. % methanol, 70
% methanol and 100% methanol were sequentially passed through each lQ to separate them into 5 fractions. That is, the water-soluble portion is classified as fraction (■), the part that elutes at a methanol concentration not exceeding 40% is classified as (■), and the part that elutes at a methanol concentration of 40% (40% part) is classified as (■) fraction, 40 to 70. % part is 1 fraction, 70-100% part is 7 fractions, and 109.
6g, 1.1g, 4.0g, 2.5g and 1.9g were obtained.

Next, the fraction (1) is passed through a column packed with silica gel, and then a mixture of chloroform 8:methanol 2.5:water species 1, chloroform 7:methanol 3:water 0.5 and methanol are sequentially passed through the column. A
Separate into fraction, B fraction, and C fraction, and about this B fraction,
The fractions eluted with 20% methanol in ODS column treatment were collected, then treated with Sephadex column, and both fractions eluted with water were treated with MCI gel CHP 20P again, and 50% methanol was passed through. Cnideoside A44 is removed by removing both solvents that are eluted.
I got stg.

In addition, the fraction eluted with 50% methanol in the above ODS column treatment was treated with Sephadex, both fractions eluted with water were treated with ODS column again, and both fractions eluted with 40% methanol were treated with Sephadex. Cnideoside B34. by removing the solvent. I got Omg.

On the other hand, after passing the above 7 fractions through a column packed with silica gel, a mixture of 9 parts chloroform: 2 parts methanol: 0.1 water, a 0.5 mixture of chloroform 7: methanol 3: water 0.5, and methanol are sequentially passed through the column. These 67 fractions were further divided into 6 fractions by ODS column (20 to 4 fractions).
0% methanol), the two fractions eluted with 30% methanol were collected, and the solvent was removed to obtain 11.5 mg of Cnideol b.

Furthermore, in the same purification process, 6-hydroxylinaroyl oxide, a monoterpene derivative, was obtained from one fraction, and 8-hydroxy-3-(S)-linalool was obtained from fraction (2).
3-0-β-D-glucopyranoside can also be obtained.

The physical properties and other characteristics of the benzofuran derivative obtained as described above are shown below.

(1) Cunideoside A (R'-H, R"-β-D-glucopyranosyl group) property
Shape: Pale yellow powder Optical rotation: [e]z7- 42.1 Cc-0,59
, H*O) thin layer chromatography (developing solvent chloroform: methanol: water = 7:3:0.5, plate ki
se1gel 60); Rf value -0,52 10% sulfuric acid spray and UV absorption: positive ultraviolet absorption spectrum (C=2.5X 10-', H2
0); UVλH20nm (log t); 283(
3,39) aX 251 (3,62) 244 (3,69) Fast atom bombardment ionization (FAB)-mass spectrum; FAB-MS m/z; 369 [M+H]”391
[M+Na]" 407[M+] From these, the molecular weight was determined to be 368.

Nuclear magnetic resonance spectrum; The structural formula below and the nuclear magnetic resonance spectrum that could be analyzed are as follows.

(i) 'H-NMR (in D20) [Aglycon] H-3= H-6: H-7= H-8: H-9= H-1Q: [Glucose] H-1': 4.77 H -2'-6': (u)'' CNMR (D20) [Aglycon] C-1: (IH, d, J = 7.0) 3.35-3.74 (6H, m) (100 MHz) δ; 128.1 7.17 7.09 7.45 6.54 2.78 2.32 (400MHz) δ; (IH,5) (IH,5) (IH,5) (LH,5) (2H,t , J = 7.3Hz) (2H,br,
d, J-7, 3Hz) C-2= C-3: C-4: C-5: C-6: C-7: C-8= C-9: C-10: C-11: [Glucose ] C-1': 102.4 C-2': 74.2 C-3': 77.3 C-4': 70.7 C-5': 77.0 C-5': 61.8 ( 2) Kunideoside B Properties: Pale yellow powder Optical rotation: [α126 153.9 122.5 154.9 123.4 99.8 146.6 107.7 28.3 39.4 184.1 19.4° ( c = 0.54.H2O) thin layer chromatography (developing solvent: chloroform: methanol: water =
7:3:0.5, plate Kiselgel 60)
; Rf value - 0,5910% sulfuric acid spray and UV absorption:
Positive ultraviolet absorption spectrum (c=2.2X 10-',)l
zo); 276 (3,38) 247 (3,75) Fast atom bombardment ionization (FAB)-mass spectrum: FAB-MS m/z; 399 [M+H]”421
[M+Na]" 439[M+] From these, the molecular weight was found to be 398.

Nuclear magnetic resonance spectrum; The structural formula below and the nuclear magnetic resonance spectrum that could be analyzed are as follows.

(l) 'HNMR (tn DzO) [Aglycone] H-6: 7-19 H-7: 7.74 H-8: 6.80 H-9: 3.06 H-10: 2.56 OMe: 4 .13 [Glucose] H-2'-6': (ii) "CNMR (DzO) [Aglycon] C-1: 127.1 3.20" 3.84 (6H, m) (100MHz) δ; (400MHz ) δ; (LH, 5) (18, d, J = 2.0 Hz) (IH, d, J = 2.0 Hz) (2H, m) (2H, m) (3H, 5) C-2: 144 .0 C-3:, 138.5 C-4: 146.5 C-5: 133. I C-6: 115.8 G-7: 146.8 C-8: 107.6 cm9: 28.0 C-10: 38.8 C-11: 181.8 OMe: 61.7 [Glucose] C -1': 105.2°C-2': 75.3 C-3': 77.7 C-4': 70.8 C-5': 77.4 C-(i': 61.8 ( 3) Cnideol b Properties: pale yellow powder thin layer chromatography (developing solvent: chloroform: methanol: water ±8:2:0.2, plate kise
lgel 60) ; Rf value -0,531O%'
fN acid spray and UV absorption: positive nuclear magnetic resonance spectrum; The structural formula and the nuclear magnetic resonance spectrum that could be analyzed are as follows.

(i) 'H-NMR (in CDCl, +CD30
D) (400MHz) δ:H-6: 6.94
(1B, 5) H-7: 7.49 (IH, d, J
-2,2Hz) H-8: 6.59 (LH,d,
J=2.2Hz) H-9: 2.91 (2H, m
)H-10: 2.58 (2H, m)OMe
: 4.02 (3H,s)(ii)”
CNMR (CDCIs + CD5OD) (100M
Hz) δ; C-1: 126.7 C-2: 144.0 C-3: 133.4 C-4: 145.8 C-5: 122. IC-6: 115.3 C-7: 144.2 C-8: 107.0 C-9: 27. I C-10: 38.3 C-11: 183.0 OMe = 61.1 Example 1 (tonic effect) Saito et al.'s method (Saito, H, and 3 others, “5tres
s ; theroleof catecholami
nes and other neurotransmi
The effect of the active ingredient of the present agent on the sexual behavior disorder in mice that occurs after stress loading was investigated in accordance with the authors' ``Pages 467-480 (1984)''.

That is, male IVOS mice with normal sexual behavior at 9 weeks of age or older and female homologous mice with normal sexual behavior at 4 weeks of age were used. All mice were preliminarily housed in private cages for one week before the start of the animal experiment, and then healthy male mice were allowed to live with female mice (10 mice per group) that had been subcutaneously administered with estradiol benzony hlOμ9/kg/day for 10 minutes. Sexual behavior training was conducted once a day for 10 days, during which time male mice that appeared to have normal sexual behavior were selected for successful intromission, and divided into groups to be equalized by the total number of successful intromission days. The animals were divided and subjected to the following tests.

Suspension stress was applied to the male mouse by suspending and fixing it upside down so that the tip of the mouse's nose was submerged in the water surface at a temperature of 20°C. This suspended state was maintained for 30 minutes on the first day of the test, and from the second day onwards, the suspended time was gradually extended by 1, and on the final day of the test, the 155th day, stress was applied for 120 minutes every day. This significantly reduced sexual behavior compared to a control group that was not subjected to stress.

First, the test was conducted every day for 10 minutes between 8pm and 9pm, with male mice from each test group cohabiting with a group of 10 acid mice, and sexual behaviors such as licking, mounting, and introductory behavior. The number of Intromissi○n movements and the time required to perform these behaviors for the first time (initial time required) were observed and measured. In other words, male IV-CS mice with normal sexual behavior that passed the above selection and 4-week-old female mice of the same species were preliminarily housed for one week, and then 10 μg/kg of estradiol benzoate was administered once every day until the day before the end of the test. The subcutaneously administered material was placed in a cage (26 x 42 x 18 cm).
) for 10 minutes to observe and measure sexual behavior (licking, mounting, introduction). When administering sex hormones to female animals, 1 mg/kg of progesterone was simultaneously administered once on the third day after the start of administration.

The above sexual behavior test will start from the next day, and the following 1.
The tests were conducted at approximately the same time for five days.

The effectiveness evaluation method is based on the number of licking, mounting, and introductions obtained through observation of the sexual behavior mentioned above, as well as the time required for the first time, and is scored in 10 stages for each.
or Index) S B I was calculated and compared and studied.

S B I = ((A+ A')X l/2) +
((B10B')X 2/21+((C+C')X3/2
) A: Score of time required until first linking A': m
Score B for the number of lickings: Score B' for the time required for the first mounting: Score C for the total number of mountings: Score C' for the time required for the first intromission: Score for the total number of intromissions. The total number of intro mission score (C') was rated as 10 regardless of the number of missions. Furthermore, the evaluation using SBI between each group is as follows:
For each group, each SBI value and the average SBI value for 15 days were calculated, and expressed as a percentage (%) with respect to day 0 for comparative study.

In addition, changes in excised organ weight and serum testosterone concentration were also evaluated.

Next, when evaluating these data, as a statistical method, all comparisons between groups are
We adopted st.

(a) Regarding sexual behavior, the average SBI percentage for 15 days is as shown in Table 1, when the SBI value on day 0 in each group is taken as 100%.

1) Percentage (%) of average SBI for days 1 to 15 when the SBI of day 08 is taken as 100 2) Normal group refers to animals that are not subjected to stress during the test period, are normally reared in individual cages, and are not given the test drug. Instead of 1% carboxymethyl cellulose sodium salt solution (hereinafter referred to as C
This is the group to which MCNa (abbreviated as MCNa) was administered.

3) Control group and other test drug administration groups were all subjected to the stress described above. However, in this control group, a 1% concentration CMCNa solution was administered instead of the test drug as in the normal group.

4) The crude extract group refers to a group to which the 50% methanol extract of Jasperum chinensis shown in Reference Example was administered as a test drug.

5) Fraction ■ group refers to a group to which the crude nideoside A and B mixed fraction shown in Reference Example was administered as a test drug.

6) Fraction B refers to a group to which the purified mixed product of Kunideoside A and B shown in the Reference Example was administered as a test drug.

7) Fraction V indicates the group to which crude Nideol b was administered.

8) A comparative example is shown in which a 50% methanol extract similarly extracted from Epimedium is used as an If quasi-drug.

9) A comparative example is shown in which yohimbine is used as a standard drug in the same manner as in 6) above.

Incidentally, the sexual behavior of male mice normally follows the sequence of licking, mounting, and introduction, and finally moves to ejaculation. Therefore, the influence or effectiveness of the device can be investigated by examining what changes occur in the sexual behavior of male mice subjected to the S-position when they coexist with female mice in heat. In other words, the more times the animal engages in sexual behavior with each of a group of 10 females within a certain period of time, the shorter the time it takes to achieve this, the more active the determined behavior is. can. Therefore, it can be determined that courageous behavior with a large SBI coefficient is active.

As is clear from Table 1, the SBI coefficient of stressed male mice clearly decreased.

From Table 1, it is clear that when the crude extract and purified product obtained from the above-mentioned production example were administered as test drugs to male mice whose sexual behavior had decreased due to stress, there was a clear improvement in sexual behavior. Is recognized. This indicates that these test drugs have the effect of improving the decline in sexual function caused by stress. It is also clear from Table 1 that the crude extract and purified product are far superior to Epimedium and yohimbine, which were used as comparative drugs.

Next, after observing the sexual behavior, the above-mentioned stress load was applied at the specified time (1:00 p.m. to 3:00 p.m.) every day, and then the test drug or 1% CMCNa solution was administered to each test group once a day. It was orally administered twice for 15 days.

Furthermore, on the final day of the test, after cervical dislocation and thoracotomy, blood was quickly collected from the heart, and the prostate, seminal vesicles, and anal muscles were removed and their weights were measured. In addition, only the serum was collected from the blood after centrifugation, and the weight of the extracted organ was measured for Radioii (b) Extracted Organ Weight, which was used as an index of sexual function improvement. The results are shown in Table 2.

This shows that the weight of the prostate and seminal vesicles clearly decreases under stress (no change in body weight was observed between the normal group and the control group); It has been found that administering the purified product obtained from the serpent roe can suppress the weight loss of the prostate and seminal vesicles caused by stress. This confirms that the test drug has the effect of improving sexual dysfunction caused by stress, since the prostate and seminal vesicles play an important role in sexual function.

(C) Regarding the serum testosterone concentration, the T in the serum
Table 3 shows the results of measuring the S concentration.

From this, it can be seen that the serum TS concentration decreases due to stress load in the control group compared to the normal group, but this decrease is significantly improved in the test drug administration group.

As described above, the agent of the present invention has a remarkable effect of improving sexual function decline caused by stress.

Example 2 (Tonic effect) The effect of the active ingredient of the agent of the present invention was investigated by causing fatigue by reducing the motor function of animals by stress loading, and measuring the recovery process of this fatigue using a rotary load.

That is, 4-week-old ddy-strain male mice were rotary loaded (axis circumference: 9.8 cm, number of rotations: 14.3).
The animals were subjected to training (times per minute), and only those that did not fall within 300 force loads were selected as experimental animals, and after a certain rest period, they were assigned to each group and subjected to the following tests.

First, the amount of movement of each animal before shaking was measured. In other words, the rotation speed of the low reload is expressed as the time (0 to 12 minutes).
changes from ((14,3 rotations/1 minute) to (28,0
rotation/1 minute)] and measured the cumulative distance walked before falling, which was scored on a 90-point scale (0 to 4).
9cm; Score 1, 50-99cm; Score 2, ... 4400cm or more: Score 90).

Note that this measurement time was interrupted at 15 minutes, and the amount of exercise of the animal corresponding to this period was treated as a score of 90. After resting for about 1 hour after completing these measurements, 1.0% concentration of CM
CNa or a suspension of the test drug in CNa was administered, and 30 minutes later, the male mice were placed in a cage (1
7X 24X 12cm), put 5 fish, 3.5c on the left and right
The sample was shaken for 3 hours in an incubator that generated 160 vibrations per minute to apply stress. This made it possible to significantly reduce motor function before shaking. Next, immediately after the end of the stress load, the amount of exercise was measured using a rotary load in the same manner as described above, and it was defined as 0 minutes. Similarly 3
The amount of exercise was measured at 0 minutes and 60 minutes, and the effects of the test drugs were compared and examined. The final tally was expressed as a percentage (%) of the momentum before shaking. The results are shown in Table 4.

From this, a clear decrease in exercise capacity was observed with shaking stress, but in the group administered the test drug, the decrease in exercise capacity due to shaking stress was significantly improved.
The test drug was found to have fatigue-relieving and tonic effects.

Example 3 (Acute toxicity test) Ddy-strain male mice (body weight 20-30 g) were kept at room temperature for 22 hours.
Preliminary breeding was carried out for one week under the conditions of temperature 1°C and humidity 55 ± 5%, healthy animals were selected, and 10 animals were added to each group to test the drug (crude extract group; 10,000 mg/kg, fraction group; 1).
1000rn/kg, fraction 8 group; 500Tng/kg,
Fraction group 7; 9 in 1000 mg/control group; 1% concentration of C
MCNa administration, dose; 0, l+x (2/mouse 109
, administration route; oral administration), and the presence or absence of death within 72 hours and simple behavioral status for approximately 60 minutes after administration.
This was carried out in accordance with the multidimensional observation method using Win. As a result, no noticeable difference in behavior from that of the control group was observed, and no deaths were observed in any of the above groups even after 72 hours. These results show that the toxicity of the agent of the present invention is extremely low even when compared with the effective doses used in Examples 1 and 2, and therefore, it can be seen that the agent of the present invention has excellent safety in use.

Formulation Example 1 (Granules) 15 g of either fraction ① or fraction B1 fraction ⋯ obtained in the course of the production example was made into a fine powder, and 1349 lactose and 1 g of magnesium stearate were mixed with this mixture. is compressed into slug tablets with a diameter of 20 mm.

This was crushed, sized, and sieved to obtain granules of 20 to 50 mesh particles.

This granule is available in the same amount from 250mg to 15% depending on the symptoms.
0 (1 mg (fraction ■, ■, B as 25 mg -15
(equivalent to 0 mg), taken 3 to 4 times a day.

Formulation Example 2 (Capsules) 20g of either fraction ① or fraction B1 fraction ⋯ obtained in the course of the production example was made into fine powder, and 95g of starch and lactose valve 809 were mixed therewith, and then , hydroxypropyl (5g
) Wet granulation is performed using an ethanol solution according to a conventional method, followed by drying and sizing to obtain granules.

Capsules were obtained by filling each capsule with 200 my of this granule.

This capsule is taken 2 to 8 capsules at a time, 3 to 4 times a day, depending on the symptoms.

Formulation example 3 (liquid for internal use) Fraction m1 obtained in the course of manufacturing example One of fractions B and V (fraction ■ to V; 0.1 to 0.3 g Fraction B; 0
．． 03~O, 1 g), glucose 2.51 g, and an appropriate amount of fragrance were added and dissolved in a 1.0% or less ethanol aqueous solution to give a total amount of 50 rnO to obtain a liquid preparation for internal use. Also, instead of the 1.0% or less ethanol mentioned above, about 14% ethanol solution was used to dissolve the solution, and the total amount was 50m0. A liquid preparation for internal use can also be obtained.

Patent applicant: Ohta Isan Co., Ltd.

Claims (1)

[Scope of Claims] 1. A highly polar extract obtained by extracting the fruit of Jasperum japonicus with an extraction solvent consisting of water or a water-containing organic solvent and then removing the extraction solvent as an active ingredient. A stress-related dysfunction improving agent. 2. The agent for improving stress-induced dysfunction according to claim 1, wherein the organic solvent is a hydrophilic solvent. 3 General formula ▲ Numerical formula, chemical formula, table, etc. ▼ (In the formula, R_1 is a hydrogen atom or a methyl group, R_2 is a hydrogen atom or a β-D-glucopyranosyl group) A stress-related dysfunction improving agent characterized by containing at least one selected from among the active ingredients. 4. The stress-induced dysfunction improving agent according to any one of claims 1 to 3, which is used as a tonic. 5. The stress-induced dysfunction improving agent according to any one of claims 1 to 3, which is used as a tonic. 6. The stress-induced dysfunction improving agent according to any one of claims 1 to 5, which is prepared for oral administration.