JP2016216364A - Bladder smooth muscle hypercontraction inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a novel bladder smooth muscle hypercontraction inhibitor.SOLUTION: The invention provides a bladder smooth muscle hypercontraction inhibitor containing Elaeagnaceae Hippophae extract as an active ingredient, the Elaeagnaceae Hippophae extract containing at least one of oleanol aldehyde, oleanolic acid, ursolic acid, pomolic acid, uvaol, a novel oleanolic acid ester compound, and isorhamnetin 7-O-rhamnoside. The invention also provides a bladder smooth muscle hypercontraction inhibitor containing Lamiaceae Rosmarinus extract as an active ingredient.SELECTED DRAWING: None

Description

  The present invention relates to a bladder smooth muscle overcontraction inhibitor. The present invention is widely used for pharmaceuticals and health foods (including foods for specified health use, functional indication foods, supplements, etc.).

  The contraction force of the smooth muscle of the bladder changes with stress. At this time, the concentration of TGF-β1 in the smooth muscle of the bladder is increased. TGF-β1 alters actin fibers in smooth muscle cells and increases the expression of stress fibers. That is, it is a cause of excessively increasing the contractile force of the bladder (overcontraction, overactivity) (Non-patent Document 1).

Overactive bladder is said to increase with age. Although physical strength and the function of various organs decrease with aging, the function of the bladder also decreases with aging.
The bladder involved in urination is subject to multiple control by three types of nerves: parasympathetic, sympathetic and somatic. The parasympathetic nerve controls the afferent and distal tracts of the bladder detrusor, and conveys urinary intention by bladder extension. The detrusor contracts with the excitement of the nerve. The sympathetic nerve mainly controls the bladder neck and is involved in maintaining urine while maintaining tension in the bladder neck. The somatic nerve controls the urogenital diaphragm and the external urethral sphincter, and is involved in voluntary contraction of the external urethral sphincter. By coordinating the movement of the bladder and urethra governed by these three types of nerves, even if a sufficient amount of urine is stored, there is no urinary incontinence and smooth urination becomes possible (Non-Patent Document 2). Therefore, urination and the autonomic nervous system are closely related, and dysuria is said to be one of the most frequent complaints of autonomic ataxia.

Overactive bladder makes urine easier. Thus, it can be seen that a drug for overactive bladder should suppress parasympathetic excitation that promotes urination. Acetylcholine is involved in the excitement of parasympathetic nerves. By using an anticholinergic agent that suppresses the action of acetylcholine, urine can be easily collected as the reverse action of promoting urination. As described above, there are solifenacin (trade name: Vesicare), propiverine (trade name: Bupfor), imidafenacin (trade name: Uritos, stabra) and the like as drugs for treating overactive bladder by anticholinergic action (Non-patent Document 3). ).

There is a β ₃ receptor stimulant as a therapeutic agent with another mechanism of action. The sympathetic and parasympathetic nerves have opposite effects, but the drug stimulates the sympathetic β ₃ receptor that controls the bladder. That is, the sympathetic nerve is stimulated in order to obtain the opposite effect of parasympathetic excitation. Thus, there is mirabegron (trade name: Betanis) as a drug for treating overactive bladder by β ₃ receptor stimulation (Non-patent Document 4).

Ramachandran A., et al. JunB mediates basal- and TGFβ1-inducedsmooth muscle cell contractility.PLoS One. 2013; 8 (1): e53430 Ryuji Sugawara, Masahiko Kishi, Yohei Tsukuzaki, Fuyuki Kanno, Tomoyuki Uchiyama, Tatsuya Yamamoto Neurology and bladder-urinary neurological mechanisms and how to treat and treat dysuria-Clinical Neurology, 53: 181-190, 2013. Satoshi Matsumoto, Hidehiro Kashizaki Considering overactive bladder medical care Characteristics, usage, and proper use of overactive bladder medicine-Based on evidence-Progress in Medicine 32 (4), 829-833 (2012). Bragg R, Hebel D, Vouri SM, Pitlick JM. Mirabegron: a Beta-3 agonist for overactive bladder.Consult Pharm. 29 (12), 823-837 (2014).

Under such a background, the present inventor has disclosed that the extract of the genus Hippophae and the genus Mannen of the family Lamiaceae, and the contraction of the collagen gel in which bladder smooth muscle cells are embedded for these components and the rat isolated bladder smooth muscle As a result of evaluating its action on acetylcholine contraction, it was found that it has an action of suppressing excessive contraction of bladder smooth muscle, and the present invention was completed.
An object of the present invention is to provide a novel bladder smooth muscle inhibitor and a urination function improving agent using the same.

The features of the present invention for solving the above-described problems are as follows.
1. Bladder smooth muscle hyperconstriction inhibitor containing as an active ingredient an extract of the genus Hippophae.
2. The Gumiaceae Hippophae extract contains at least one of oleanolaldehyde, oleanolic acid, ursolic acid, pomolic acid, ubaol, compound A represented by the following chemical formula (1) and isorhamnetin 7-O-rhamnoside. Certain above 1. The bladder smooth muscle overcontraction inhibitor described in 1.
3. An inhibitor of bladder smooth muscle overcontraction by TGF-β1 containing an extract of the genus Hippophae as an active ingredient.
4). The above-mentioned Gumiaceae Hippophae extract contains at least one of oleanolaldehyde, oleanolic acid, ursolic acid, pomolic acid, ubaol, the above compound A and isorhamnetin 7-O-rhamnoside. The bladder smooth muscle overcontraction inhibitor described in 1.
5. 2. The above 3, which comprises at least one of oleanolaldehyde, oleanolic acid, ursolic acid, pomolic acid, ubaol, the above compound A, and isorhamnetin 7-O-rhamnoside as an active ingredient. The bladder smooth muscle overcontraction inhibitor described in 1.
6). Bladder smooth muscle hyperconstriction inhibitor with acetylcholine, which contains an extract of the genus Hippophae genus as an active ingredient.
7). 6. An extract of the genus Hippophae belonging to the family Gumiaceae, which contains at least one of ursolic acid, pomolic acid, compound A and isorhamnetin 7-O-rhamnoside. An inhibitor of bladder smooth muscle overcontraction with acetylcholine according to 1.
8). A bladder smooth muscle hyperconstriction inhibitor by acetylcholine containing at least one of ursolic acid, pomolic acid, compound A and isorhamnetin 7-O-rhamnoside as an active ingredient.
9. Compound A represented by the following chemical formula (1).
10. Bladder smooth muscle hyperconstriction inhibitor containing an extract of the genus Mannenro of Lamiaceae as an active ingredient.
11. 10. The ursolic acid is contained in the extract of the Lamiaceae Mannenrou genus. The bladder smooth muscle overcontraction inhibitor described in 1.

2 is a scheme showing a method for isolating oleanolaldehyde, oleanolic acid, ursolic acid, pomolic acid, ubaol, and compound A from seaberry. 1 is a scheme showing a method for isolating isorhamnetin 7-O-rhamnoside from sea berry. It is a graph which shows the TGF-b1-induced bladder smooth muscle cell-containing collagen gel contraction inhibitory effect of Seaberry extract. It is a graph which shows the TGF-b1-induced bladder smooth muscle cell-containing collagen gel contraction inhibitory effect of ursolic acid. It is a graph which shows the effect | action with respect to the shrinkage | contraction of the collagen smooth gel containing TGF-b1 induction | guidance | derivation of saw palm oil (comparative example). It is a graph which shows the inhibitory effect with respect to TGF-b1-induced bladder smooth muscle cell containing collagen gel contraction of a seaberry extract containing component. It is a graph which shows the effect | action with respect to the acetylcholine contraction of the rat isolated bladder smooth muscle in a seaberry extract. It is a graph which shows the effect | action with respect to the acetylcholine contraction of the rat isolated bladder smooth muscle in ursolic acid. It is a graph which shows the effect | action with respect to the acetylcholine contraction of the rat isolated urinary bladder smooth muscle in pomol acid, isorhamnetin 7-O-rhamnoside, and compound A (3-O-p-coumaroyl-2,19,23-trihydroxy oleanolic acid). It is a graph which shows the effect | action with respect to the acetylcholine contraction of the rat isolated bladder smooth muscle in a rosemary extract. FIG. 3 is a diagram showing a correlation between HMBC and ¹ H- ¹ H COZY of Compound A.

Hereinafter, the present invention will be described in detail.
In addition, the present invention is characterized in that an extract of the genus Hippophae is used as an active ingredient. Examples of the plant belonging to the genus Hippophae include, for example, sea berries (Hippophae rhhamnoides L.), salicifolia (Hippophae salifolia L.), tibetana (Hippophae thibetana L.), and p.
Among them, the use of sea berries (Hippophae ramnoides L.) is most preferable from the viewpoint of the content of the active ingredient in the obtained extract.

When preparing an extract of the genus Hippophae plant, there is no particular limitation on the extraction target part of the plant, and for example, an appropriate part such as a seed, a fruit, a leaf, a bark, a trunk, a root, or an above-ground part may be used. Of these, fruits are the most preferred. As the extraction method, if necessary, the extraction target sites can be washed with water, dried, chopped or pulverized in advance, and then brought into contact with an extraction solvent according to a conventional method such as an immersion method or a countercurrent extraction method.

Examples of the extraction solvent for the extract include alcohols such as methanol, ethanol, propanol and butanol; glycols such as 1,3-butylene glycol, glycerin and propylene glycol; esters such as ethyl acetate and butyl acetate; Ethers such as propyl ether, isopropyl ether, tetrahydrofuran and dioxane; halogenated hydrocarbons such as dichloromethane and chloroform; ketones such as acetone, methyl ethyl ketone, diethyl ketone and cyclohexanone; use hexane, cyclohexane, petroleum ether, water, etc. Can do. Of these, ethanol, 1,3-butylene glycol, water, acetone, dichloromethane and the like are preferably used. These solvents can be used alone or in admixture of two or more. When mixing and using a solvent, what is necessary is just to adjust the mixing ratio of each solvent suitably according to the kind of solvent.

The extraction method is not particularly limited, and after adding a solvent (for example, ethanol) to the fruit of the genus Hippophae, heat is heated to a degree that does not deactivate the active ingredient contained in the extract. An extraction method, a supercritical extraction method, or the like can be applied as appropriate. Moreover, various well-known extraction methods, such as the immersion extraction method which carries out batch processing by immersing the fruit of the genus Hippophae in a fixed amount of solvent, and the continuous extraction method which continues sending a solvent continuously, can be applied.

An example of a specific extraction method is, for example, about 5 to 50 times the dry weight, preferably about 10 to 40 times the extraction solvent, and immersed in the genus Hippophae. The active ingredient can be extracted by heating (25 to 100 ° C.) and stirring the solvent for about 1 to 6 hours. Of course, the type of solvent, the amount of solvent, the heating temperature, the heating time, etc. may be appropriately adjusted so that the active ingredient can be extracted efficiently.
Moreover, it is preferable that the extract of the genus Hippophae contains at least one of oleanolaldehyde, oleanolic acid, ursolic acid, pomolic acid, ubaol, compound A, and isorhamnetin 7-O-rhamnoside.

Further, the bladder smooth muscle hyperconstriction inhibitor of the present invention comprises at least one of oleanolaldehyde, oleanolic acid, ursolic acid, pomolic acid, ubaol, compound A, and isorhamnetin 7-O-rhamnoside as an active ingredient. Features. The method for obtaining oleanolaldehyde, oleanolic acid, ursolic acid, pomolic acid, ubaol, compound A, and isorhamnetin 7-O-rhamnoside is not particularly limited,
It is preferable to extract and isolate from the fruit of the genus Hippophae. This is because these compounds are contained at a high concentration. In addition, these compounds can be isolated and purified based on, for example, the methods of Examples in the present specification.

The bladder smooth muscle overcontraction inhibitor of the present invention contains an extract of the genus Mannenro of Lamiaceae as an active ingredient. Rosemary is particularly preferable as the genus Mannenrou.
There are no particular limitations on the site to be extracted from the plant, and suitable parts such as seeds, fruits, flowers, leaves, bark, trunks, roots, and above-ground parts can be used, but leaves are the most preferable. As the extraction method, if necessary, the extraction target sites can be washed with water, dried, chopped or pulverized in advance, and then brought into contact with an extraction solvent according to a conventional method such as an immersion method or a countercurrent extraction method.

Examples of the extraction solvent for the extract include alcohols such as methanol, ethanol, propanol and butanol; glycols such as 1,3-butylene glycol, glycerin and propylene glycol; esters such as ethyl acetate and butyl acetate; Ethers such as propyl ether, isopropyl ether, tetrahydrofuran and dioxane; halogenated hydrocarbons such as dichloromethane and chloroform; ketones such as acetone, methyl ethyl ketone, diethyl ketone and cyclohexanone; use hexane, cyclohexane, petroleum ether, water, etc. Can do. Of these, ethanol, 1,3-butylene glycol, water, acetone, dichloromethane and the like are preferably used. These solvents can be used alone or in admixture of two or more. When mixing and using a solvent, what is necessary is just to adjust the mixing ratio of each solvent suitably according to the kind of solvent.

The extraction method is not particularly limited, and after adding a solvent (for example, ethanol) to the genus Ganoderaceae, it is heated and heated to such an extent that the activity of the active ingredient contained in the extract is not inactivated. Alternatively, a supercritical extraction method or the like can be applied as appropriate. Moreover, various well-known extraction methods, such as the immersion extraction method which carries out batch processing by immersing Labiatae in the fixed amount of solvent, the continuous extraction method which continues sending a solvent, etc., are applicable.

An example of a specific extraction method is, for example, about 5 to 50 times by weight, preferably about 10 to 40 times by weight of an extraction solvent, and dipped in a genus Lamaceae. The active ingredient can be extracted by heating (25 to 100 ° C.) and stirring the solvent for about 1 to 6 hours. Of course, the type of solvent, the amount of solvent, the heating temperature, the heating time, etc. may be appropriately adjusted so that the active ingredient can be extracted efficiently. It is preferable that an extract of the genus Labiatae belongs to ursolic acid. At this time, the content of ursolic acid is not particularly limited, but it is preferably 0.5% or more, preferably 1.5% or more, and more preferably 3.0% or more.
Furthermore, when using a rosemary extract as an extract of Lamiaceae Mannenrou, a commercially available product may be used. Specifically, “rosemary (rosemary extract)” manufactured by Sabinsa may be used. Incidentally, “Rosemary (Rosemary extract)” manufactured by Sabinsa Co., Ltd. contains 1.5% or more of ursolic acid.

The ratio of the bladder smooth muscle overcontraction inhibitor of the present invention is not particularly limited as long as the bladder smooth muscle overcontraction inhibitor exhibits the bladder smooth muscle overcontraction inhibitory action. Moreover, it may consist only of a bladder smooth muscle overcontraction inhibitor. Specific examples of the content of the bladder smooth muscle overcontraction inhibitor include 0.005% by weight or more in terms of the total amount in terms of dry weight. In the case of using an extract of the genus Hippophae belonging to the family as an inhibitor of bladder smooth muscle overcontraction, the amount of the compound or the like contained therein may be used.

The shape of the bladder smooth muscle overcontraction inhibitor of the present invention is not particularly limited. Examples thereof include powder, granule, tablet, pill, emulsion, liquid, suspension, concentrated extract, and dry extract.

The bladder smooth muscle overcontraction inhibitor of the present invention can be applied to various uses for the purpose of the above-mentioned effects. Applications include pharmaceuticals, reagents (for research and test kits), and compositions for oral consumption (for example, health foods, functional labeling foods, supplements, foods for specific uses, and supplements for pets).

The pharmaceutical composition containing the bladder smooth muscle overcontraction inhibitor of the present invention may contain an excipient, a carrier or an additive. The excipient, carrier and additive are not particularly limited as long as they are usually used and pharmaceutically acceptable, and the type and composition thereof can be appropriately changed.

Examples of the excipient include sodium chloride and sodium citrate, and examples of the carrier include sterilized water, physiological saline, and various buffers. Examples of additives include thickeners, buffer materials, preservatives, preservatives, and the like.

The dosage form of the pharmaceutical composition is not particularly limited and may be appropriately selected as necessary. For example, oral preparations such as tablets, capsules, granules, fine granules, powders; injections And parenterals such as suppositories and coating agents.

Oral preparations such as tablets, capsules, granules, fine granules, powders and the like are produced according to a conventional method using, for example, starch, lactose, sucrose, trehalose, mannitol, carboxymethylcellulose, corn starch, inorganic salts, The compounding quantity of the compound of this invention in these formulations is not specifically limited, It can set suitably. In this type of preparation, binders, disintegrants, surfactants, lubricants, fluidity promoters, corrigents, colorants, fragrances and the like can be used as appropriate.

In the case of a parenteral preparation, the dose is adjusted according to the age, weight, disease level, etc. of the patient, and for example, intravenous administration, intravenous infusion, subcutaneous injection, intramuscular injection or the like is used. This parenteral preparation is produced according to a conventional method, and distilled water for injection, physiological saline and the like can be generally used as a diluent. Furthermore, you may add a bactericidal agent, antiseptic | preservative, a stabilizer, etc. as needed. In addition, from the viewpoint of stability, this parenteral preparation can be frozen after filling into a vial or the like, the water can be removed by ordinary freeze-drying treatment, and the liquid preparation can be re-prepared from the freeze-dried product immediately before use. Further, if necessary, an isotonic agent, a stabilizer, an antiseptic, a soothing agent and the like may be added. Examples of other parenteral agents include liquid preparations for external use, coating agents such as ointments, suppositories for rectal administration, etc., and these are also produced according to conventional methods.

When the present invention is used for health foods (including functional indication foods, foods for specified health use, health drinks and supplements), the compound of the present invention is added to the health foods as raw materials for various health foods, or dextrin as necessary It can be processed into pellets, tablets, granules, etc. together with excipients such as lactose and starch, fragrances, pigments, etc., or coated with gelatin and formed into capsules to produce health foods.

The mixing ratio of the bladder smooth muscle overcontraction inhibitor blended in the health food is not particularly limited as long as the expected effect of the bladder smooth muscle overcontraction inhibitor is obtained. Is about 0.0001 to 20 mg. In the case of using an extract, it is sufficient to set an amount such that the compound (1) contained therein has the above ratio.

These health foods can be blended with various components depending on the type, for example, glucose, fructose, sucrose, maltose, sorbitol, stevioside, corn syrup, lactose, citric acid, tartaric acid, malic acid, Succinic acid, lactic acid, L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid ester, polyglycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, gum arabic Food materials such as carrageenan, casein, gelatin, pectin, agar, vitamin Bs, nicotinic acid amide, calcium pantothenate, amino acids, calcium salts, pigments, fragrances and preservatives can be used. In addition, bladder smooth muscle hyperconstriction inhibitors with health maintenance functions include other antioxidants and health food ingredients such as antioxidants (reduced ascorbic acid (vitamin C), vitamin E, reduced) Type glutatin, tocotrienol, vitamin A derivatives, lycopene, lutein, astaxanthin, zeaxanthin, fucoxanthin, uric acid, ubiquinone, coenzyme Q10, folic acid, garlic extract, allicin, sesamin, lignans, catechin, isoflavone, chalcone, tannins, flavonoids , Coumarin, isocoumarins, blueberry extract), health food ingredients (V. (vitamin) A, V.B1, V.B2, V.B6, V.B12, VC, VD, VE, VP, choline, niacin, pantothene) Acid, calcium folate, EPA, oligosaccharide, dietary fiber, squalene, soybean lecithin, taurine, dona Ella, protein, octacosanol, DHA, egg yolk lecithin, linoleic acid, lactoferrin, magnesium, zinc, chromium, selenium, potassium, heme iron, oyster meat extract, chitosan, chitin oligosaccharide, collagen, chondroitin, turmeric, licorice, kokushi, kehi , Hawthorn, ginger, ganoderma, swordfish extract, turtle, licorice, wolfberry, keihi, hawthorn, ginger, ganoderma, plantain, chamomile, chamomile, dandelion, hibiscus, honey, boren, royal jelly, lime, lavender, rosehip, bifido Fungus, faecalis, lacris, wheat germ oil, sesame oil, perilla oil, soybean oil, medium chain fatty acid, agaricus, ginkgo biloba extract, turmeric, chondroitin, brown rice germ extract, litchi, onion, DHA, EPA, DPA, green tea, winter Summer grass, garlic, bee, papaya, puer, propolis, mugwort, yabushitake, royal jelly, saw palmetto, hyaluronic acid, collagen, gabba, harp seal oil, shark cartilage, glucosamine, lecithin, phosphatidylserine, paddy carrot , Mulberry leaves, soy extract, echinacea, sorghum, barley extract, olive leaf, olive fruit, gymnema, banaba, salacia, garcinia, chitosan, st jones wort, jujube, carrot, passion flower, broccoli, placenta, pearl barley, grape seeds , Peanut seed coat, bilberry, black cohosh, maria thistle, laurel, raffa, black vinegar, bitter gourd, maca, safflower, flax, oolong tea, flower spine, caffeine, capsaicin, xylooligosaccharide, glucosamine, buckwheat, Truss, dietary fiber, protein, prunes, spirulina, barley young leaves, nucleic acid, yeast, shiitake mushroom, plum meat, amino acids, deep sea potato extract, noni, oyster meat, suppon, champignon, plantain, acerola, pineapple, banana, peach, apricot , Melon, strawberry, raspberry, orange, fucoidan, mesimacob, cranberry, chondroitin sulfate, zinc, iron, ceramide, silk peptide, glycine, niacin, chesttree, L-cysteine, red wine leaf, millet, horsetail, biotin, center Asiatica, Haskap, Pycnogenol, Fuuki, Rhubarb, Clove, Catechin, Puer, Citric Acid, Beer Yeast, Merirot, Black Zinger, Ginger, Gadju, Nattokinase, Benikouji, Tocotrienol, Lactoferi , Cinnamon, buckwheat, cocoa, yuzu seed extract, perilla seed extract, lychee seed extract, evening primrose extract, black rice extract, alpha-lipoic acid, raw coffee bean extract, Unshu mandarin orange extract, triterpenoid, kiwi seed extract, red ginger extract , Astaxanthin, Walnut extract, Resveratrol, Red rice extract, White fungus polysaccharide, Strawberry seed extract, Strawberry seed extract, Leek seed extract, Lingonberry extract, Cherry blossom extract, Sasakurehito yotake extract, Makiberry extract, Black ginger extract , Rice polyamine, wheat polyamine, Junsai extract, purple tea extract, chrysanthemum flower extract) and the like.

Examples of the present invention will be described below.
Example 1. Preparation of Seaberry (Hippophae rhamnoides L.) extract Seaweed (Hippophae rhamnoides L.) 5 times the weight of the dried and crushed dried fruits of n-hexane at 20 to 40 ° C., 1 hour stirring After that, the product was degreased and solid-liquid separation was performed to obtain a degreased raw material.
Then, this degreased raw material is dried, and extracted with 70 volumes of 70 vol% aqueous ethanol at 70 ° C or higher for 1 hour with stirring, followed by solid-liquid separation with a centrifuge. The extract was concentrated to dryness to obtain a seaberry extract.

2. Isolation of components of seaberry extract (1) Among components of seaberry extract, oleanolaldehyde, oleanolic acid, ursolic acid, pomolic acid, ubaol, and compound A represented by the above chemical formula (1) are shown in FIG. Isolated according to the scheme shown. That is, dried fruit of Hippophae ramnoides L. was pulverized and pulverized fruit (364 g) was extracted with 50 wt% aqueous methanol under stirring for 2 hours to obtain an extract. Thereafter, this extract was subjected to liquid-liquid partitioning (n-hexane, ethyl acetate, butanol, and water), an n-hexane layer (0.73 g), an ethyl acetate layer (6.0 g), a butanol layer (4.3 g), and an aqueous layer ( 13.2 g) was obtained.
Then, the n-hexane layer and the ethyl acetate layer are mixed and separated by a silica gel column (see FIG. 1 for conditions such as a mobile phase). Fractions 1-8 were obtained. In HPLC, Fr. From the two fractions, oleanolaldehyde (9 mg), Fr. From the three fractions, ursolic acid (34 mg), Fr. From the four fractions, pomolic acid (7 mg), ursolic acid (205 mg) and a novel substance (23 mg), Fr. From 5 fractions, pomolic acid (14 mg), ubaol (35 mg) and a new substance (5 mg) were obtained.

(3) ¹ H and ¹³ C-NMR spectra were measured for the new substance. The data is shown below.
I) Physicochemical data Pale yellow powder. ¹ H-NMR (CD ₃ OD, 600 MHz) d _H : 0.77 (3H, s, 24-H), 0.79 (3H, s, 26-H), 0.92 (3H, s, 29-H), 0.93 (3H, s, 30-H), 1.01 (3H, s, 25-H), 1.20 (3H, s, 27-H), 2.50 (1H, brs, 18-H) , 3.88 (2H, m, 23-H), 3.92 (1H, m, 2-H), 4.93 (1H, m, 3-H), 5.30 (1H, brs, 12-H), 6.40 (1H, d , J = 5.5, 8'-H), 6.79 (1H, d, J = 8.9, 3 ', 5'-H), 7.45 (1H, d, J = 8.2, 2', 6'-H), 7.65 (1H, d, J = 8.2, 7'-H). 13C-NMR (CD ₃ OD, 150 MHz) d _C : 13.9 (C-24), 14.0 (C-26), 16.0 (C-29), 16.0 (C-30), 16.9 (C-25), 26.0 (C-27) 42.5 (C-14), 43.7 (C-1), 48.1 (C-9), 54.5 (C-18), 64.5 ( C-23), 67.1 (C-2), 73.0 (C-19), 79.4 (C-3), 115.2 (C-8 '), 116.3 (C-3', 5 '), 123.5 (C-1 '), 128.6 (C-12), 130.6 (C-2', 6 '), 139.6 (C-13), 146.0 (C-7'), 160.7 (C-4 '), 169.4 (C-9' ), 181.7 (C-28).
II) Two-dimensional NMR
^{HMQC (1 H-detected multiple quantum} coherence spectrum) and ^{HMBC (1 H-detected multi-} bond heteronuclear multiple quantum coherence spectrum), 1 H- 1 H COSY (1 H- 1 H correlation spectroscopy) results of the analysis of 11 Correlation between HMBC and ¹ H- ¹ H COZY shown in FIG. From the above, it was confirmed that the novel substance was the compound A represented by the chemical formula (1).

(2) Isoramnetin 7-O-rhamnoside was isolated according to the scheme of FIG. That is, the above butanol layer was subjected to Fr. using a silica gel column (refer to FIG. 1 for conditions such as a mobile phase). Separated into 1B-6B fractions. Of these, Fr. From the 3B fraction, isorhamnetin 7-O-rhamnoside (6.4 mg) was obtained with RP-HPLC (TSK-Gel ODS-120T) mobile phase 60% MeOH.

3. Preparation of rosemary extract 100g of rosemary leaves and stems were crushed and extracted with 70 volumes of 70vol% hydrous ethanol at 70 ℃ or higher for 1 hour with stirring, and then solid-liquid separation was performed with a centrifuge. This extract was concentrated to dryness to obtain 7.6 g of rosemary extract. This rosemary extract contained 0.98% ursolic acid.

Test Example 1: Evaluation of bladder smooth muscle overcontraction inhibitory action (evaluation of contractile action of collagen gel embedded with bladder smooth muscle cells)
Human bladder smooth muscle cell fluid (1.5 × 10 ⁶ cells / mL) and rat tail-derived collagen fluid (1.2 mg / mL) were mixed at a ratio of 1: 9, and seeded at 500 uL in a 24-well plate. After incubation for 1 hour, 500 μL of 0.5% FCS-containing medium was added to each well and incubated overnight. Here, TGF-b1 (2.5 ng / mL) and an evaluation sample were added and cultured for 24 hours, and then the gel was taken out. After imaging in PBS, the gel area was calculated. The results are shown in FIG. 3 (seaberry extract), FIG. 4 (ursolic acid) and FIG.

Results and results of test of Example 1 in Test Example 1, Saw Palmetto Oil (Indena Co., Ltd.), known for its urinary dysfunction-inhibiting effect, in which seaberry extract (Fig. 3) and ursolic acid (Fig. 4) have been shown to suppress gel contraction As a result of evaluation in the same manner, almost no contraction inhibitory action was observed (FIG. 5). It was confirmed that the seaberry extract has the effect of suppressing chronic bladder overcontraction due to stress.
Further, as shown in FIG. 6, as a result of the same evaluation of the components contained in the seaberry extract, triterpenes showed ubaol, pomolic acid, oleanolaldehyde, and a novel substance (compound A; 3-Op-coumaroyl-2,19 , 23-trihydroxy oleanolic acid) was found to have a contractile inhibitory effect. The flavonoid isorhamnetin 7-O-rhamnoside was also inhibited.

Test Example 2: Effects on acetylcholine-induced contraction of isolated rat urinary bladder smooth muscle In rats, the bladder smooth muscle was suspended in a Magnus device and stimulated with carbachol (a parasympathomimetic agent) in nutrient solution. Examined. Specifically, 3 x 5 mm sections were prepared from the bladder removed from SD rats (male, 7-8 weeks old) and suspended in a Magnus tank containing Krebs solution. While a gas mixture of 95% oxygen / 5% carbon dioxide was vented, a load of 1 g was applied at 37 ° C. The tension waveform was recorded via an isotonic transducer, and after stabilization, 10 ^-6 M carbachol was added to induce contraction. After confirmation of shrinkage, washing and stabilization, an evaluation sample was added and allowed to act for 10 minutes. Thereafter, 10 ^-8 to 10 ^-4 M carbachol was cumulatively added, and the contraction waveform was recorded. Finally, lavage and confirmation of contraction with 10 ^-4 M carbachol were performed to confirm that the bladder smooth muscle was not toxic.
The obtained results were graphed by calculating the contraction rate of each carbachol concentration with respect to the maximum contraction caused by carbachol. The results are shown in FIG. 7 (seaberry extract), FIG. 8 (ursolic acid), FIG. 9 (other seaberry constituents), and FIG. 10 (rosemary extract).

Results and Effects of Examples in Test Example 2 As shown in FIG. 7, the seaberry extract exhibited an inhibitory action on carbachol contraction of 10 ⁻⁶ M or less. Further, as shown in FIG. 8, the action of ursolic acid as a main component was clearer, and carbachol contraction of 3 × 10 ⁻⁶ M or less was suppressed in the range of 1 to 100 ug / mL. In addition, as shown in FIG. 9, it showed an inhibitory action on carbachol contraction of isorhamnetin 7-O-rhamnoside 3 × 10 ⁻⁵ M or less. It was significantly suppressed at 10 ^-7 M. In addition, Compound A (10 μg) showed a tendency to suppress 10 ⁻⁴ M carbachol contraction, and pomolic acid suppressed 3 × 10 ⁻⁶ M to 3 × 10 ⁻⁵ M carbachol contraction. The results revealed that seaberry extract and ursolic acid inhibit bladder contraction by acting on bladder smooth muscle. Furthermore, as shown in FIG. 10, in the rosemary extract, 10 ⁻⁵ M carbachol contraction was suppressed in the range of 10 to 100 ug / mL. As a result, it became clear that rosemary extract acts on bladder smooth muscle to suppress bladder contraction.

The compounding example of the bladder smooth muscle excessive contraction inhibitor (Gumiaceae Hippophae genus extract) by this invention is shown. In addition, the following compounding examples do not limit the present invention. Moreover, although not mentioned in the compounding example, it can mix | blend similarly in a rosemary extract.
Formulation Example 1: Chewing gum (functional indication food or food for specified health use)
53.0wt% sugar
Gum base 20.0
Glucose 10.0
Minamata 16.0
Fragrance 0.5
Gummy Hiphopafae Extract 0.5
100.0wt%

Formulation Example 2: Gummy (Functional indication food or food for specified health use)
Reduced water tank 40.0wt%
Granulated sugar 20.0
Buto sugar 20.0
Gelatin 4.7
Water 9.68
Yuzu fruit juice 4.0
Yuzu Flavor 0.6
Dye 0.02
Gummyaceae Hippophae Extract 1.0
100.0wt%

Formulation example 3: Candy (functional indication food or food for specified health use)
50.0 wt% sugar
Minamata 33.0
Water 14.4
Organic acid 2.0
Fragrance 0.2
Gummy Hiphopafae Extract 0.4
100.0wt%

Formulation Example 4: Yogurt (hardware / soft) (functional food or food for specified health use)
Milk 41.5wt%
Nonfat dry milk 5.8
Sugar 8.0
Agar 0.15
Gelatin 0.1
Lactic acid bacteria 0.005
Gummy Hiphopafae Extract 0.4
Perfume
Water residue
100.0wt%

Formulation Example 5: Soft drink (functional display food or food for specified health use)
Fructose glucose liquid sugar 30.0wt%
Emulsifier 0.5
Gummy Hippophae Extract 0.3
Perfume
Purified water residue
100.0wt%

Formulation Example 6: Tablet confectionery (functional display food or food for specified health use)
76.4 wt% sugar
Glucose 19.0
Sucrose fatty acid ester 0.2
Gummy Hiphopafae Extract 0.5
Purified water 3.9
100.0wt%

Formulation Example 7: Soft capsule (supplement or medicine)
Brown rice germ oil 47.0wt%
Yuzu seed oil 40.0
Emulsifier 12.0
Gummyaceae Hippophae Extract 1.0
100.0wt%

Formulation Example 8: Tablet (supplement or medicine)
Lactose 54.0wt%
Crystalline cellulose 30.0
Starch degradation product 10.0
Glycerin fatty acid ester 5.0
Gummyaceae Hippophae Extract 1.0
100.0wt%

  As described above, the present invention is useful as a bladder smooth muscle hyperconstriction inhibitor and can be used for pharmaceuticals, health foods and the like.

Claims (11)

Bladder smooth muscle hyperconstriction inhibitor containing as an active ingredient an extract of the genus Hippophae. The Gumiaceae Hippophae extract contains at least one of oleanolaldehyde, oleanolic acid, ursolic acid, pomolic acid, ubaol, compound A represented by the following chemical formula (1) and isorhamnetin 7-O-rhamnoside. The bladder smooth muscle overcontraction inhibitor according to claim 1.
An inhibitor of bladder smooth muscle overcontraction by TGF-β1 containing an extract of the genus Hippophae as an active ingredient. 4. The gummy family Hippophae extract comprises at least one of oleanolaldehyde, oleanolic acid, ursolic acid, pomolic acid, ubaol, the compound A and isorhamnetin 7-O-rhamnoside. Bladder smooth muscle hyperconstriction inhibitor. The bladder smooth muscle overcontraction inhibitor according to claim 3, comprising at least one of oleanolaldehyde, oleanolic acid, ursolic acid, pomolic acid, ubaol, the compound A, and isorhamnetin 7-O-rhamnoside as an active ingredient. Bladder smooth muscle hyperconstriction inhibitor with acetylcholine, which contains an extract of the genus Hippophae genus as an active ingredient. The urinary urinary hippophae extract contains at least one of ursolic acid, pomolic acid, compound A and isorhamnetin 7-O-rhamnoside, and is acetylcholine-induced bladder according to claim 6 Smooth muscle overcontraction inhibitor. A bladder smooth muscle hyperconstriction inhibitor by acetylcholine containing at least one of ursolic acid, pomolic acid, compound A and isorhamnetin 7-O-rhamnoside as an active ingredient. Compound A represented by the following chemical formula (1).
Bladder smooth muscle hyperconstriction inhibitor containing an extract of the genus Mannenro of Lamiaceae as an active ingredient. 11. The bladder smooth muscle hyperconstriction inhibitor according to claim 10, wherein the extract of the genus Lamaceae contains ursolic acid.