JP2003277274A - Agent having female hormone-like action - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a food, a medicine or the like which has a female hormone(estrogen)-like action and is useful as an antiosteoporotic agent, as a menopausal syndrome-improving or -treating agent, as an agent for preventing or treating a prostate cancer, a prostatomegaly, a breast cancer, or a uterine cancer, or as an agent for improving or treating an acne of a female postpuberty sideration. <P>SOLUTION: The agent having a female hormone(estrogen)-like action, the antiosteoporotic agent, the menopausal-improving or -treating agent, the agent for preventing or treating a prostate cancer, a prostatomegaly, a breast cancer, or a uterine cancer, the agent for improving or treating an acne of a female postpuberty sideration, the food, the medicine, and the cosmetics are characterized by containing a propolis, a kale, or an extract thereof. <P>COPYRIGHT: (C)2004,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an anti-osteoporosis agent having a female hormone (estrogen) -like action, an agent for improving / treating menopausal disorders, an agent for preventing / treating prostate cancer / prostatic hypertrophy / breast cancer and uterine cancer, or a woman. The present invention relates to an agent for ameliorating and / or treating acne after puberty, and an oral composition, an external composition for skin or food, a medicine, and a cosmetic containing the same.

[0002]

2. Description of the Related Art Estrogen is a kind of female hormone, and is a general term for hormones having an estrus effect and substances having a similar effect. Estradiol is the major naturally occurring steroidal estrogen, but other steroidal estrogens such as estrone, estriol, ericin, homoestrone and ethinyl estradiol are known.
Further, as non-steroidal estrogen, for example, diethylstilbestrol, hexestrol and the like are known.

Of these estrogen-acting substances, steroidal estrogens are used as medicines for indications such as amenorrhea, anovulatory cycle, functional uterine bleeding, and uterine growth failure due to a decrease in estrogen. It is also used for the treatment of menopausal disorders and the suppression of lactation. In addition, nonsteroidal estrogens are mainly used as therapeutic agents for diseases such as prostate cancer and benign prostatic hyperplasia.

On the other hand, it is known that plants also contain estrogen acting substances, which are collectively called phytoestrogens. As such phytoestrogens, it has been reported that isoflavones such as genistein, daidzein and myloestrol exhibit relatively strong estrogenic activity (Chiharu Umehara, Takeo Sato, steroid hormones, clinical physiology of pharmaceuticals). , Estrogen, published by Nankodo, 1966). More recently, daily intake of plants containing these phytoestrogens has led to
It has been pointed out that it may contribute to a decrease in the expression of diseases such as osteoporosis and gynecological diseases, breast cancer, uterine cancer, prostate cancer, and benign prostatic hyperplasia, and further a decrease in blood cholesterol level.

More recently, among the metabolites of certain microorganisms, 5,7-dihydroxy-2- (4-hydroxyphenyl) -3-methylchroman-4-one has been isolated and found to be estrogenic. It has been reported to have activity (JP-A-2-218678 and JP-A-2-3).
No. 00181).

[0006] All the currently used estrogen agonists have a problem that they have side effects such as gastrointestinal disorders, thrombosis, uterine bleeding, and liver disorders. It is known that there is. Therefore, an estrogen agonist in which these side effects are reduced is desired.

Menopause, on the other hand, is defined as a syndrome of endocrine, somatic and physiological alterations that occur at the end of the reproductive period in women. Irregular menstruation is the manifestation of prolonged menstrual bleeding caused by loss of ovulation. Loss of ovulation is caused by impaired follicular development. Recently,
The most common method for the treatment of climacteric disorders is hormone replacement, including the administration of birth control pills, orally with estrogen and progesterone formulations or with progesterone only formulations (Shaaban, 1996.
Year). While alleviating the symptoms of menopause, these treatments present many associated risks and side effects. Risks associated with hormone therapy include endometrial cancer, hypertension, hyperlipidemia, gallstones (gallstones), breast cancer, and deep vein thrombosis (Barentsen, 1996).

On the other hand, acne vulgaris (acne) develops only after puberty, so that modulation of endocrine function associated with increased secretion of sex hormones is closely related to the development of this disease. it is obvious. In other words, it is the basis that the amount of androgen secreted by both men and women is relatively higher than that of estrogen (Fragrance Journal, No. 74, p. 4-7, 1985). Although acne is said to occur frequently during adolescence, recently, the number of female acne patients who develop after adolescence is increasing. It has been reported that a large number of these women have elevated blood levels of androgens such as testosterone and dihydrotestosterone (Journal of Japanese Cosmetic Science 21: 4, 337-340 1
997). Thus, female acne that develops after adolescence involves hormonal imbalance.

[0009] Such acne treatment includes antibiotics,
Internal medicines such as vitamins and hormones, and external medicines containing salicylic acid, sulfur and the like having a keratolytic action, a bactericidal action and an anti-inflammatory action are prescribed. However, like the premenstrual syndrome, it is performed under the supervision of a doctor, and it involves the annoyance of having to visit the doctor. In addition, vitamin B ₂ · B ₆ preparations and external medicines having bactericidal and anti-inflammatory effects are commercially available as internal medicines to which acne is applied as general medicines, but none of them pay attention to hormone balance.

On the other hand, in recent years, the problem of osteoporosis has become serious. Osteoporosis is a condition in which bone becomes brittle due to a decrease in bone mass.

In normal bone, bone resorption (dissolution of the pelvis) and bone formation are alternately carried out in a well-balanced manner, and a constant bone mass is maintained while undergoing metabolic turnover. However, when this balance is lost and absorption is promoted, the bone weight decreases, the bone becomes thin and the bone is prone to fracture, and pain may occur. Such a condition is called osteoporosis, and it is a characteristic feature especially in postmenopausal women. Although it varies according to statistics, it is also reported to be observed in about 1/4 of women of this age. In addition, it can cause the elderly to be bedridden, so the quality of life in an aging society (Quality of Life)
From the viewpoint of improvement of life), highly effective therapeutic agents are required. Furthermore, since the treatment is long-term, high safety is also required.

[0012] Although the mechanism of osteoporosis has not been completely clarified, it is considered necessary to suppress bone resorption or promote bone formation during treatment. Based on this idea, estrogen has been clinically introduced for the treatment of osteoporosis.

Estrogen has both a bone resorption suppressing action and a bone formation promoting action, and suppresses the progression of osteoporosis. However, during long-term administration, in addition to gastrointestinal symptoms such as abdominal bloating and nausea, the occurrence of breast and endometrial cancers, endometrial bleeding, increased swelling and breast pain, which are peculiar to female hormones, are observed. Serious side effects may occur,
Side effects such as abnormal lipid metabolism and venous thrombosis have also been observed. Therefore, there is a problem in safety in long-term administration. As described above, in spite of the desire to develop safe foods and medicines having excellent preventive or therapeutic effects on osteoporosis, at present, no products have been developed that have both efficacy and safety.

[0014]

The object of the present invention is to provide a medicine or food which has an estrogen-like action and is sufficiently satisfactory for safe and reliable treatment even after long-term administration or application, It is to provide cosmetics.

[0015]

Means for Solving the Problems As a result of intensive studies to solve the above-mentioned problems, the present inventor has found that it has been used as a dietary supplement in Japan for the purpose of anticancer action and immunostimulatory action. The present invention has been completed by discovering that propolis which is present in the plant, and kale of the family Cruciferae, which is commonly called and drunk as green juice, have an estrogenic effect.

That is, the present invention relates to an estrogen-like agent containing propolis or kale or an extract thereof, an anti-osteoporosis agent, an agent for improving / treating menopausal disorders, prevention of prostate cancer / prostatic hypertrophy / breast cancer and uterine cancer / Therapeutic agents, post-pubertal onset acne improving / therapeutic agents, oral compositions containing at least one of these agents, external compositions for skin, and further,
These compositions are foods, medicines, cosmetics.

[0017]

BEST MODE FOR CARRYING OUT THE INVENTION The propolis used in the present invention is
It is a resinous black mass collected by bees, Brazilian propolis, Chinese propolis, Australian propolis,
Uruguay-produced propolis, Japan-produced propolis, etc. may be of any origin, and is not particularly limited,
From the viewpoint of versatility, Brazilian propolis and Chinese propolis are preferable.

Propolis is a dried product obtained by drying propolis itself, a pulverized product thereof, a supercritical extract, a crude extract with water or an organic solvent such as alcohol, ether or acetone, and a crude extract, column chromatography, etc. It includes all of the extract fractions obtained by stepwise purification by various types of chromatography. These may be used alone or in combination of two or more.

For example, the extract obtained by adding 3 L of a 99.5% ethanol extract to 1 kg of a dried raw product of propolis from Brazil and immersing it overnight at room temperature may be used as it is, or may be subjected to various chromatographies. You may use it, combining and refine | purifying.

The concentration of the propolis extract in the solution of the extracted propolis extract is not particularly limited, but is preferably 15 to 70% by weight, preferably about 20 to 60% by weight. If this concentration is 15% by weight or less, it is necessary to evaporate a large amount of a solution such as ethanol or water at the time of drying.
If it exceeds the above range, the viscosity of the solution becomes too high, which may deteriorate the processability.

The kale (Brassicca Ol) used in the present invention
eracea L.var.acephala DC.) has kitchen kale,
Malo kale, bush kale, tree kale, collard, green algae and so on. The cruciferous plant is originally a vegetable originally from Southern Europe and is said to be the original cabbage species. The leaves may be those usually used for food, and the cultivation method and cultivation place are not particularly limited.

As the kale and its extract, a dried product obtained by drying the kale itself, a crushed product thereof, a squeezed juice, a crude extract with water or an organic solvent such as alcohol, ether or acetone, and a crude extract are distributed, It includes all of the extract fractions obtained by stepwise purification by various chromatography such as column chromatography. These may be used alone or in combination of two or more.

For example, 3 L of 99.5% ethanol extract may be added to 1 kg of dried matter such as leaves, stems, flowers and roots of kale, and the extract obtained by immersing at room temperature overnight may be used as it is. The extract may be purified by combining various chromatographies.

It has never been known in the past that the dried product or extract of propolis and kale according to the present invention has an estrogen-like effect, which is a new finding obtained by the present invention.

The propolis and kale according to the present invention have an excellent estrogenic action, and are anti-osteoporotic agents, agents for improving and treating menopausal disorders, and agents for preventing / treating prostate cancer / prostatic hypertrophy / breast cancer and uterine cancer. Alternatively, it can be used as a food, a medicine, or a cosmetic for the purpose of improving and / or treating acne for postpubertal onset in women.

The propolis and kale of the present invention are used as an estrogen-like agent, an anti-osteoporosis agent, an agent for improving / treating menopausal disorders, an agent for preventing / treating prostate cancer / prostatic hypertrophy / breast cancer and uterine cancer, or after puberty in women. It can be produced as a food or a medicine containing an acne improving / therapeutic agent for onset.

Estrogen-like agents, anti-osteoporosis agents, agents for improving / treating menopausal disorders, agents for preventing / treating prostate cancer / prostatic hypertrophy / breast cancer and uterine cancer, or agents for improving / treating acne after puberty in women As a method of applying such a medicine, either oral administration or parenteral administration can be adopted. Upon administration, the active ingredient can be mixed with a solid or liquid non-toxic pharmaceutical carrier suitable for administration methods such as oral administration, rectal administration and injection, and then administered in the form of a conventional pharmaceutical preparation. As such a formulation, for example,
Examples include solid agents such as tablets, granules, powders and capsules, solutions such as solutions, suspensions and emulsions, and freeze-dried preparations, and these preparations can be prepared by a conventional preparation method. Examples of the nontoxic carrier for pharmaceutical use include glucose, lactose, sucrose, starch, mannitol,
Examples thereof include dextrin, fatty acid glyceride, polyethylene glycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitan fatty acid ester, amino acid, gelatin, albumin, water and physiological saline. In addition, if necessary, a stabilizer, a wetting agent,
Conventional additives such as an emulsifier, a binder and a tonicity agent can be added as appropriate.

[0028] As foods, as they are, or by adding various nutritional components or by being contained in foods and drinks, anti-osteoporosis agents, agents for improving and treating menopausal disorders, prostate cancer, benign prostatic hyperplasia, breast cancer and uterine cancer. It is eaten as a health food or a food material useful as a preventive / therapeutic agent or an agent for improving / treating acne of females after puberty. For example, after adding the above-mentioned suitable auxiliaries, using a conventional means, a form suitable for food, such as granules, granules, tablets, capsules, paste, etc. may be provided for food, Further, various foods, for example, ham, processed meat products such as sausage, fish products such as kamaboko, chikuwa, bread, confectionery, butter, milk powder, and used by adding to fermented dairy products, water, fruit juice, milk, You may use it, adding it to drinks, such as tea and a soft drink.

The effective doses of the propolis and kale of the present invention are appropriately selected and determined according to the age, body weight, symptom of the patient, degree of the patient, administration route, administration schedule, formulation form, strength of inhibitory activity of the material, etc. However, for example, in the case of oral administration of propolis, it is generally 10 to 500 mg per day.
/ kg body weight, preferably about 150 to 350 mg / kg body weight per day, and may be administered in several divided doses per day. For kale, the dry weight is usually 0.1 g / body weight k in terms of adults
It is more than g and may be administered in several divided doses per day.

When the propolis or kale of the present invention is used as a cosmetic or a cosmetic material containing an estrogen-like agent, for example, a dried product or an extract of Chinese propolis is added to wheat germ oil or olive oil. The composition containing an estrogen-like agent can be added to the composition and used as a cosmetic material. The amount of the dried product or extract of kale of the present invention is not particularly limited, but as an example, 0.1% by weight or more based on the weight of wheat germ oil or olive oil 6
It is suitable to be 0% by weight or less, preferably 0.5% by weight or more and 50% by weight or less. In the case of propolis, 0.01% by weight or more and 60% by weight or less, preferably 0.1% by weight or more and 50% by weight or less is suitable for the weight of wheat germ oil or olive oil.

Further, the above estrogen-like agent-containing composition can be directly used as a cosmetic ingredient to produce a cosmetic having an estrogen-like effect. Cosmetics include oils and fats such as vegetable oils, higher fatty acids, higher alcohols, silicones, anionic surfactants, cationic surfactants, amphoteric surfactants, nonionic surfactants, preservatives,
Sugars, sequestering agents, polymers such as water-soluble polymers, thickeners, powder components, UV absorbers, UV blockers,
A moisturizing agent such as hyaluronic acid, a fragrance, a pH adjusting agent, a desiccant and the like can be contained. Other medicinal and physiologically active ingredients such as vitamins, skin activating agents, blood circulation promoting agents, indigenous bacteria control agents, active oxygen scavengers, anti-inflammatory agents, anti-cancer agents, whitening agents and bactericides can also be contained.

As the cosmetics, skin cosmetics such as lotion, emulsion, cream and pack, makeup base lotion, makeup cream, emulsion or cream-like or ointment type foundation, lipstick, eye color,
It can be used as makeup cosmetics such as blush color, body creams such as hand creams, leg creams and body lotions, bath salts, oral cosmetics and hair cosmetics. As the cosmetic containing the estrogen-like agent-containing composition obtained by the method of the present invention, from the viewpoint of function, for example, emulsion, cosmetics, face cream, hand cream, lotion, essence, shampoo, rinse are preferable. .

Such cosmetics can be manufactured by a conventional method. The amount of the dry product or extract of the propolis of the present invention in the cosmetic is not particularly limited, but as an example, 0.01% by weight or more and 20% by weight or less of the total weight of the cosmetic is suitable. .

Since the propolis and kale of the present invention are natural products, their toxicity is low. For example, the ethanol extract of kale was orally administered to rats for a long period of 1000 mg / kg daily for 100 days. No change was observed and no change in body weight was observed.

[Embodiment] The embodiment will be specifically described below, but the invention is not limited thereto. Production Example 1 (Production of propolis extract) Propolis original masses from Brazil and China are 10 g each for 99.
After adding 3 L of 5% ethanol and immersing it overnight at 50 ° C., ethanol was removed by a rotary evaporator to obtain 324 mg and 216 mg of propolis extract, respectively.

Production Example 2 (Production of Kale Extract) Kale leaves were dried at 90 ° C. to deactivate the enzyme which is a bitter and astringent ingredient. 20 liters of 99.5% ethanol (Wako Pure Chemical Industries) while refluxing 4 kg of the crushed product with an electric water bath
80 g of kale extract was obtained.

Test Method The binding test to the rat uterine estrogen receptor was evaluated by B.
KOFFMAN et al. (J. Steroid Biochem. Molec. Bio
l., 38 (2) 135-139, 1991) with some modifications.

[Preparation of cytosol (cytosol) fraction] Unless otherwise specified, all the operations described below were carried out at a temperature of 4 ° C.

First, four times the amount of phosphate buffer (pH 8.0) was added to crushed frozen uterine tissue, and a Polytron PT-10 probe (manufactured by Brinkman Instrument Co. Inc.) was used under full control conditions. Homogenization was performed by crushing for 5 seconds, cooling in ice for 60 seconds until the next crushing, and then crushing for 5 seconds twice. 1050 this homogenate
After centrifugation at 00 × g for 30 minutes, the obtained supernatant was decanted and used as a cytosolic fraction. The protein content of the cytosolic fraction was measured by the method of Lowry et. Al. (J. Biol. Chem. 193 265-275 1951) using bovine serum albumin as a protein standard sample.

[Evaluation of Binding Test to Rat Uterus Estrogen Receptor] The cytosolic fraction prepared so as to have a protein concentration of 1.2 to 7.2 mg / ml was dissolved in a phosphate buffer (pH 8.0) to give 1 nM [ ^[3 H] -labeled 17-β-estradiol was added to each propolis extract or kale extract obtained in Production Examples 1 and 2 at 500 μg / ml, and the mixture was reacted at 40 ° C. for 20 hours. In addition, non-specific binding test (blank)
Was performed in the presence of a 3000-fold excess of distilbesterol.

Next, the released steroids and sample samples were treated with a dextran-treated activated carbon suspension (0.5% Norrit A, 0.005% dextran-containing phosphate buffer solution (pH
7.4); hereinafter abbreviated as DCC), and removed by incubation at 4 ° C. for 10 minutes. Then 50
DCC was precipitated by centrifuging at 00 × g for 10 minutes, and the resulting supernatant containing protein-bound steroid 100
μl scintillation cocktail solution (Packard Instr
ument Co., Downer Grove, III) 4ml was added and scintillation counter (Packard Model 2425, 30-40%
efficiency). The inhibition rate was calculated by the following formula. Inhibition rate (%) = 100 - _{[(C 1 -B) / (C} 0 -
B)] × 100 (wherein C ₁ is a known amount of the test compound and [ ³ H] -labeled 17-β.
- ^[3 H] labeled in a state where estradiol coexist
Represents the amount of 17-β-estradiol bound to the rat uterine estrogen receptor, C ₀ represents the amount of [ ³ H] -labeled 17-β-estradiol bound to the rat uterine estrogen receptor when the test compound was excluded, B is [ ^{3 in the} presence of excess distilbestrol (3 × 10 ^-6 M)]
[H] Labeled amount of 17-β-estradiol bound to rat uterine estrogen receptor. ) The results are shown in Table 1.

[0042]

[Table 1]

The IC ₅₀ value of 17-β-estradiol as a positive control in this assay system was 0.
It was 84 nM (2.29 × 10 ⁻⁴ ng / ml). As can be seen from Table 1, the propolis extract and kale extract of the present invention antagonize the estrogen receptor binding inhibitory action of 17-β-estradiol and have a strong estrogen-like action.

Example 1 [Production of tablet] Using the ethanolic extract of propolis from Brazil obtained in Production Example 1, a tablet having the following composition was produced according to a conventional method. (Composition) (Composition: wt%) Brazilian Propolis Extract 24 Lactose 63 Corn Starch 12 Guar Gum 1

Example 2 [Production of juice] Using the ethanolic extract of kale obtained in Production Example 2, a juice having the following composition was produced according to a conventional method. (Composition) (Composition: wt%) Frozen concentrated Unshu mandarin orange juice 5.0 5.0 Fructose glucose liquid sugar 11.0 Citric acid 0.2 L-Ascorbic acid 0.02 Perfume 0.2 Pigment 0.1 Kale extract 0. 2 Water 83.28

Example 3 [Production of Face Cream] Using the ethanol extract of Chinese propolis obtained in Production Example 1, a face cream having the following composition was produced according to a conventional method. (Composition) (Composition: wt%) Isopropyl isostearate 8.0 Jojoba oil 6.0 Cetanol 8.0 Stearyl alcohol 2.0 Polyoxyethylene lauryl ether 1.5 Propylene glycol 6.0 Sorbitol 1.0 Paraben 0.0 4 Chinese Propolis Extract 0.5 Vitamin E 0.5 Perfume 0.1 Purified Water 66.0

[0047]

EFFECT OF THE INVENTION Since the propolis and kale of the present invention have a female hormone (estrogen) -like action, foods, pharmaceuticals or cosmetics containing them are anti-osteoporosis agents,
Improvement / treatment of menopausal disorders, prostate cancer / prostatic hypertrophy /
It is useful as a prophylactic / therapeutic agent for breast cancer and uterine cancer or an ameliorating / therapeutic agent for post-pubertal acne in women.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61K 35/78 A61K 35/78 C A61P 13/08 A61P 13/08 15/00 15/00 15/12 15 / 12 17/10 17/10 19/10 19/10 35/00 35/00 F term (reference) 4B018 MD48 MD78 ME05 ME06 ME08 ME10 ME14 MF02 4C083 AA081 AA082 AA111 AA122 AC072 AC122 AC132 AC182 AC342 AC482 AD662 CC02 DD31 EE14 4C087 AA01 AA02 BB22 CA06 CA47 MA17 MA28 MA35 MA52 MA63 NA09 NA10 NA14 ZA81 ZA89 ZA97 ZB26 4C088 AB15 AC03 AC05 AC11 BA10 CA06 MA17 MA28 MA35 MA52 MA63 NA09 NA10 NA14 ZA81 ZA89 ZA97 ZB26

Claims (9)

[Claims] 1. A female hormone-like agent containing propolis, kale, or an extract thereof. 2. An anti-osteoporosis agent containing propolis, kale or an extract thereof. 3. An agent for improving and / or treating menopausal disorders, which comprises propolis, kale, or an extract thereof. 4. A prophylactic / therapeutic agent for prostate cancer / prostatic hypertrophy / breast cancer and uterine cancer, which comprises propolis, kale or an extract thereof. 5. A remedy for acne after puberty, which contains propolis, kale or an extract thereof. 6. An oral composition containing the agent according to claim 1. 7. A composition for external use for skin, which comprises the agent according to claim 1. 8. The composition according to claim 6, which is a food or a medicine. 9. The composition according to claim 7, which is a cosmetic or a medicine.