JP2012158574A - Estrogen-like agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an estrogen-like agent for preventing/treating osteoporosis, gynecology disease, prostate cancer, prostatic hyperplasia or the like.SOLUTION: The estrogen-like agent includes a compound represented by general formula (I) [wherein, Ar denotes a C6 to C14 aryl group which may be substituted with one to three substituents selected from the group consisting of a halogen atom, a hydroxy group and a 1C to 6C alkoxy group; A denotes a 1C to 4C alkylene group, a 2C to 4C alkenylene group or a 2C to 4C alkynylene group which may be substituted; and Rdenotes a hydrogen atom, a 1C to 6C alkyl group which may be substituted, a 3C to 6C cycloalkyl group which may be substituted or a 6C to 14C aryl group which may be substituted] or its salt.

Description

  The present invention relates to an estrogenic agent containing a plant-derived compound of the following formula (I) or a salt thereof. Furthermore, the present invention relates to the estrogen-like agent for treatment / prevention of osteoporosis, gynecological disease, prostate cancer, prostatic hypertrophy, etc., and a pharmaceutical and a food containing the estrogen-like agent.

  Estrogens are a type of steroid hormone synthesized from cholesterol, and lack of estrogen increases the possibility of climacteric disorder and osteoporosis. In particular, postmenopausal women suddenly decrease estrogen secretion, and it is necessary to artificially incorporate estrogen into the body. At present, hormone replacement therapy (HRT) is actively performed, but there is data that it becomes easy to get uterine cancer and breast cancer by taking a large amount of estrogen, and the right or wrong is questioned.

  In recent years, phytoestrogens such as isoflavones contained in soybeans have attracted attention. Phytoestrogens have estrogenic effects when estrogen is deficient, and suppress estrogen when estrogen is in excess. Therefore, phytoestrogens are thought to be an alternative to HRT.

  Much research has been done to search for phytoestrogens. For example, coffee extract is known to have an estrogenic action, but its active ingredient has not been elucidated (Non-patent Document 1). In addition, caffeic acid ester, which is one of the components of coffee extract, includes high retroviral agents (Patent Documents 1 and 2), α-glucosidase inhibitors (Non-Patent Document 2), and antioxidants (Non-Patent Document 3). Is not known for estrogenic activity.

European Patent Publication No. 1166782 Japanese Patent Laid-Open No. 2002-020283

Journal of Toxicology and Environmental Health, 20: 37-49, 1987 Biosci. Biotechnol. Biochem. 74 (4), 741-745, 2010 Biosci. Biotechnol. Biochem. 60 (7), 1093-1095, 1996

  The object of the present invention is to provide plant-derived estrogenic agents, pharmaceuticals and foods that are used for the treatment and prevention of osteoporosis, gynecological diseases, prostate cancer or benign prostatic hyperplasia, etc., with low risk of uterine cancer and breast cancer There is to do.

  As a result of diligent research to solve the above-mentioned problems, the present inventors have found that estrogen is contained in an ethyl acetate soluble fraction of a methanol extract of the cruciferous family Arahana which is a traditional vegetable in Shiga Prefecture. An estrogen-like agent comprising the component (compound of formula (I) described below) useful for the treatment and prevention of osteoporosis, gynecological disease, prostate cancer, prostatic hypertrophy, etc. The inventors have found pharmaceuticals and foods and have completed the present invention.

That is, the present invention is as follows.
[1] General formula (I):

[Wherein Ar is a C6-C14 aryl group optionally substituted with 1 to 3 substituents selected from the group consisting of a halogen atom, a hydroxy group and a C1-C6 alkoxy group, and A is substituted. An optionally substituted C1-C4 alkylene group, an optionally substituted C2-C4 alkenylene group or an optionally substituted C2-C4 alkynylene group, R ¹ is a hydrogen atom, an optionally substituted C1-C6 An alkyl group, an optionally substituted C3-C6 cycloalkyl group or an optionally substituted C6-C14 aryl group], or an estrogenic agent containing the salt thereof.
[2] Ar is phenyl optionally substituted by 1 to 3 substituents selected from the group consisting of a halogen atom, a hydroxy group and a C1-C6 alkoxy group, A is a C2-C4 alkenylene group, and R ¹ The estrogenic agent according to [1], wherein is a hydrogen atom or a C1-C6 alkyl group.
[3] The compound represented by the formula (I) or a salt thereof is at least one selected from the group consisting of methyl caffeate, ethyl caffeate, propyl caffeate and butyl caffeate. The estrogenic agent described.
[4] The estrogen-like agent according to any one of [1] to [3], wherein the compound represented by the formula (I) or a salt thereof is derived from Suruga greens.
[5] The estrogenic agent according to any one of [1] to [4], which is used for treatment and / or prevention of osteoporosis or gynecological disease.
[6] The estrogenic agent according to any one of [1] to [4], which is used for treatment and / or prevention of prostate cancer or benign prostatic hyperplasia.
[7] A pharmaceutical comprising the estrogen-like agent according to any one of [1] to [6].
[8] A food containing the estrogen-like agent according to any one of [1] to [6].
[9] The food according to [8], wherein the food is a health functional food or a dietary supplement.
[10] The food according to [9], wherein the health functional food is a food for specified health use or a nutritional functional food.

  According to the present invention, since the compound of the formula (I) specifically shows activity on estrogen β cells existing in a wide range in the body, the estrogen-like agent of the present invention is not only used for reproductive organs but also for bone. Estrogen-like action can be exerted in a wide range in the body such as brain and brain. In addition, since the compound of formula (I) also has an antioxidant action, it is useful for the treatment and prevention of arteriosclerosis and cancer. Furthermore, since the active ingredient is derived from a natural product called Suruga greens, it can be used safely for a long time and can be used as an alternative to HRT.

FIG. 1 shows a scheme for extracting active ingredients from Suruga vegetables. FIG. 2 shows the estrogenic activity (α cell) of the fraction obtained by subjecting the ethyl acetate soluble fraction (EtOAc ext) of Suruga greens to column chromatography. FIG. 3 shows the estrogenic activity (β cells) of the fraction obtained by subjecting the ethyl acetate soluble fraction (EtOAc ext) of Suruga greens to column chromatography. FIG. 4 shows the estrogenic activity (α cell) of caffeic acid ester. The horizontal axis shows the relative luminescence intensity when the luminescence intensity when the activity test is performed by adding only the solvent (ethanol) is 1 (negative control). FIG. 5 shows the estrogenic activity (β cells) of caffeic acid ester. The horizontal axis shows the relative luminescence intensity when the luminescence intensity when the activity test is performed by adding only the solvent (ethanol) is 1 (negative control).

  Hereinafter, the present invention will be described in detail.

  The present invention is directed to general formula (I):

[Wherein Ar is a C6-C14 aryl group optionally substituted with 1 to 3 substituents selected from the group consisting of a halogen atom, a hydroxy group and a C1-C6 alkoxy group, and A is substituted. An optionally substituted C1-C4 alkylene group, an optionally substituted C2-C4 alkenylene group or an optionally substituted C2-C4 alkynylene group, R ¹ is a hydrogen atom, an optionally substituted C1-C6 An alkyl group, an optionally substituted C3-C6 cycloalkyl group or an optionally substituted C6-C14 aryl group], or an estrogen-like agent (hereinafter referred to as this). Also referred to as an estrogenic agent of the invention).

  Examples of the “halogen atom” in the present specification include fluorine, chlorine, bromine and iodine.

  Examples of the “C1-C6 alkoxy (group)” in the present specification include methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, isopentyloxy, neopentyloxy Tert-pentyloxy, hexyloxy, 1-ethylpropoxy and the like.

Examples of the “C6-C14 aryl (group)” in the present specification include phenyl, naphthyl (eg, 1-naphthyl, 2-naphthyl), anthryl, phenanthryl, etc., preferably phenyl or naphthyl, More preferred is phenyl.
The “C1-C4 alkylene (group)” in the present specification may be either linear or branched, for example, —CH ₂ —, — (CH ₂ ) ₂ —, — (CH _{2 ) 3 -, - (CH 2} ) 4 -, - CH (CH 3) -, - C (CH 3) 2 -, - CH (CH 3) -CH 2 -, - CH 2 -CH (CH 3) - _{, -C (CH 3) 2 -CH} 2 -, - CH 2 -C (CH 3) 2 - as a "C2-C4 alkenylene (group)" is in this specification and the like, straight-chain or branched For example, -CH = CH-, -CH = CH (CH ₂ )-, -CH = C = CH-, -CH = CH (CH ₂ ) _2- , -CH ₂ CH = CH (CH ₂ )-and the like.
The “C2-C4 alkynylene (group)” in the present specification may be either linear or branched, for example, —C≡C—, —C≡C (CH ₂ ) —, — C≡C (CH ₂ ) ₂ —, —CH ₂ —C≡C—CH ₂ — and the like.
Examples of the “C1-C6 alkyl (group)” in the present specification include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, tert-pentyl and hexyl. 3,3-dimethylpropyl and the like, preferably a C1-C4 alkyl group.
Examples of the “C3-C6 cycloalkyl (group)” in the present specification include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

In the present specification, “optionally substituted C1-C4 alkylene group”, “optionally substituted C2-C4 alkenylene group”, “optionally substituted C2-C4 alkynylene group”, “substituted” Examples of the “optionally substituted C1-C6 alkyl group”, “optionally substituted C3-C6 cycloalkyl group” and “optionally substituted C6-C14 aryl group” include, for example,
(1) a halogen atom,
(2) a hydroxy group,
(3) an amino group,
(4) Nitro group,
(5) a cyano group,
(6) C1-C6 alkyl group,
(7) C1-C6 alkoxy group,
(8) C3-C6 cycloalkyl group,
(9) C6-C14 aryl group,
(10) a heterocyclic group;
“C1-C4 alkylene group”, “C2-C4 alkenylene group”, “C2-” each optionally substituted with 1 to 5, preferably 1 to 3 substituents selected from the group consisting of Examples include “C4 alkynylene group”, “C1-C6 alkyl group”, “C3-C6 cycloalkyl group” and “C6-C14 aryl group”.

In the present specification, unless otherwise specified, for example, as a ring-constituting atom, one or two kinds selected from the group consisting of a nitrogen atom, a sulfur atom and an oxygen atom in addition to a carbon atom are used. A 4 to 14 membered (preferably 5 to 10 membered) (monocyclic, bicyclic or tricyclic) heterocyclic group containing 1 to 4 heteroatoms, preferably (i) 5 to 14 membered (preferably 5 To 10-membered) aromatic heterocyclic group, and (ii) 4- to 10-membered (preferably 5- to 10-membered) non-aromatic heterocyclic group.
Examples of the “aromatic heterocyclic group” include furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, imidazolyl, pyrazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,3, 4-oxadiazolyl, furazanyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, tetrazolyl, pyridyl, Monocyclic aromatic heterocyclic groups such as pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl; for example, benzofuryl, isobenzofuryl, benzo [b] thienyl, indolyl, isoindolyl, 1H-indazolyl, benzimidazolyl, benzoxazolyl, benzo [d ] Isoxazoli , Benzothiazolyl, benzo [d] isothiazolyl, 1H-benzotriazolyl, quinolyl, isoquinolyl, cinnolinyl, quinazolinyl, quinoxalinyl, phthalazinyl, naphthyridinyl, purinyl, pteridinyl, carbazolyl, α-carbolinyl, β-carbolinyl, γ-carbolinyl, acridinyl, Phenoxazinyl, phenothiazinyl, phenazinyl, phenoxathiinyl, thiantenyl, phenatridinyl, phenanthrolinyl, indolizinyl, pyrrolo [1,2-b] pyridazinyl, pyrazolo [1,5-a] pyridyl, imidazo [1,2-a Pyridyl, imidazo [1,5-a] pyridyl, imidazo [1,2-a] pyridazinyl, imidazo [1,2-a] pyrimidinyl, 1,2,4-triazolo [4,3-a] pyridyl, , 2,4-triazolo [4,3-b] aromatic fused heterocyclic group pyridazinyl, and the like.
Examples of the “non-aromatic heterocyclic group” include azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, tetrahydrofuryl, thiolanyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl, thiazolidinyl, piperidyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, piperazinyl and the like Non-aromatic heterocyclic group; isochromanyl, dihydrobenzopyranyl, isochromenyl, chromenyl (2H-chromenyl, 4H-chromenyl), 1,2,3,4-tetrahydroisoquinolyl, 1,2,3,4-tetrahydroquino Non-aromatic condensed heterocyclic groups such as ril, 2,3-dihydrobenzofuranyl, benzo [1,3] dioxolyl are exemplified.

The “optionally substituted C6-C14 aryl group” represented by Ar is preferably substituted with 1 to 3 substituents selected from the group consisting of a halogen atom, a hydroxy group, and a C1-C6 alkoxy group It may be phenyl. More preferred is phenyl optionally substituted with 1 to 3 substituents selected from the group consisting of a hydroxy group and a C1-C6 alkoxy group, and more preferred is substitution with 1 to 3 hydroxy groups Optionally substituted phenyl, particularly preferably phenyl substituted with 1 to 3 hydroxy groups, particularly preferably phenyl substituted with 2 hydroxy groups.
A is preferably a C1-C4 alkylene group, a C2-C4 alkenylene group or a C2-C4 alkynylene group, more preferably a C2-C4 alkenylene group.
R ¹ is preferably an optionally substituted C1-C6 alkyl group, more preferably a C1-C6 alkyl group, still more preferably a C1-C4 alkyl group.

As a preferred embodiment of the compound represented by the formula (I),
Ar is phenyl optionally substituted by 1 to 3 substituents selected from the group consisting of a halogen atom, a hydroxy group and a C1-C6 alkoxy group, A is a C2-C4 alkenylene group, and R ¹ is hydrogen An atom or a C1-C6 alkyl group, more preferably, a compound in which Ar is a phenyl substituted with a hydroxy group, A is a C2-C4 alkenylene group and R ¹ is a C1-C6 alkyl group, or a salt thereof.

  Specific examples of particularly suitable compounds represented by the formula (I) include methyl caffeate, ethyl caffeate, propyl caffeate, and butyl caffeate.

  As the compound represented by the formula (I) or a salt thereof, any of chemical synthesis methods, natural products derived from animals and plants, those obtained by fermentation methods or gene recombination methods may be used.

  The starting material for the production of the compound represented by the formula (I) or a salt thereof is described in J. Med. Chem. 1997, 40, 1186-1194, Chem. Biol. Interaction, 84 (1992) 277-290, etc. It can be prepared by the methods described and known processes. That is, it can be produced by applying various known synthesis methods utilizing the characteristics based on the basic skeleton of the compound represented by formula (I) or the type of substituent. For example, alkylation, acylation, amination, imination, halogenation, reduction, oxidation, condensation and the like can be mentioned, and reactions or methods usually used in the art can be used.

  Examples of the salt of the compound represented by the formula (I) include pharmacologically acceptable salts such as trifluoroacetic acid, acetic acid, lactic acid, succinic acid, maleic acid, tartaric acid, citric acid, and gluconic acid. , Ascorbic acid, benzoic acid, methanesulfonic acid, p-toluenesulfonic acid, cinnamic acid, fumaric acid, phosphonic acid, hydrochloric acid, nitric acid, hydrobromic acid, hydroiodic acid, sulfamic acid, sulfuric acid, etc. Acid addition salts; for example, metal salts such as sodium, potassium, magnesium, calcium; for example, salts with organic bases such as trimethylamine, triethylamine, pyridine, picoline, N-methylpyrrolidine, N-methylpiperidine, N-methylmorpholine, etc. Is mentioned.

  When the compound represented by the formula (I) has an isomer such as an optical isomer, a stereoisomer, a positional isomer, and a rotational isomer, these are also contained as the compound represented by the formula (I). In addition, each can be obtained as a single product by a synthesis method and a separation method known per se. For example, when an optical isomer exists in the compound represented by the formula (I), the optical isomer resolved from the compound is also included in the compound represented by the formula (I). Here, the optical isomer can be produced by a method known per se.

The compound represented by the formula (I) may be a crystal.
The crystal of the compound represented by the formula (I) can be produced by crystallization by applying a crystallization method known per se to the compound represented by the formula (I). When the compound represented by the formula (I) is a crystal, it is included in the compound represented by the formula (I) whether the crystal form is single or a crystal form mixture.

The compound represented by the formula (I) may be labeled with an isotope (eg, ³ H, ¹⁴ C, ³⁵ S, ¹²⁵ I, etc.) and the like.
Furthermore, the compound represented by the formula (I) may be a hydrate, a non-hydrate, a non-solvate or a solvate.
Further, a deuterium converter obtained by converting ¹ H to ² H (D) is also included in the compound represented by the formula (I).

An embodiment in which the compound represented by the formula (I) or a salt thereof (sometimes referred to as a compound of the present invention) is derived from Suruga vegetables is also included in the present invention.
Suruga greens are cruciferous vegetables that are Shiga's traditional vegetables, and are native to the Suruga indigenous to the Suruga in Tsuchiyama Town, Koka City, Shiga Prefecture. The compound represented by the above formula (I) derived from Suruga vegetable is obtained from Suruga vegetable as follows.

  Suruga greens are usually sown in mid-October, and the stalks and leaves in the center of the strains erected from March to April are edible. Extract and concentrate Suruga rape leaves with methanol to obtain a methanol extract. The obtained methanol extract is subjected to solvent fractionation successively with hexane and ethyl acetate. The obtained ethyl acetate-soluble fraction is fractionated with a column packed with ODS, and the fraction showing estrogenic activity is further subjected to conventional purification and the like to obtain the compound of the present invention.

  Since the compound of the present invention has an estrogenic action, the estrogenic agent containing the compound of the present invention can be used in mammals (eg, humans, monkeys, cats, pigs, horses, cows, mice, rats, guinea pigs, dogs). , Rabbits, etc.) are useful for the treatment or prevention of diseases involving estrogen, particularly for the prevention, improvement or treatment of diseases or symptoms resulting from decreased estrogen or hormonal imbalance.

  Examples of diseases or symptoms resulting from decreased estrogen or hormonal balance include osteoporosis, gynecological diseases, lipid metabolism abnormalities, and the like. Examples of gynecological diseases include climacteric disorder, juvenile climacteric disorder, amenorrhea, menstrual abnormalities, menstrual syndrome, breast cancer, and anovulatory uterine bleeding.

  Examples of lipid metabolism abnormalities include hyperlipidemia, arteriosclerotic diseases such as myocardial infarction, angina pectoris, and cerebral infarction.

  Of these, the estrogenic agent of the present invention is suitable for the treatment and / or prevention of osteoporosis and menopause.

  The estrogen-like agent of the present invention is suitable for hormone replacement therapy (HRT) for estrogen reduction.

  Furthermore, since estrogen has an action of suppressing the action of male hormones, the estrogen-like agent of the present invention can be applied mainly to prostate cancer, prostatic hypertrophy, etc. whose growth is promoted by testosterone, a male hormone. Especially, it is suitable for the treatment and / or prevention of prostate cancer and prostatic hypertrophy.

The estrogenic agent of the present invention has an activity specific to estrogen receptor (ER) β cells.
In order for estrogen to act, binding to the estrogen receptor (ER) is required. There are two types of ER, ERα and ERβ. ERα is mainly expressed in the liver and reproductive organs, and ERβ is expressed not only in the reproductive organs but also in a wide range in the body such as bones and brains. Therefore, the estrogen-like agent of the present invention having an activity specific to β cells is effective for symptoms and diseases caused by a decrease in estrogen and hormonal malfunction in a wide range of the body.

  Since the compound of the present invention has an antioxidant action, the estrogenic agent of the present invention can be applied to the prevention and treatment of arteriosclerotic diseases such as myocardial infarction, angina pectoris, and cerebral infarction, and cancer.

  A medicament containing the estrogenic agent of the present invention is also encompassed by the present invention (hereinafter also referred to as the medicament of the present invention).

  The pharmaceutical of the present invention can be administered orally or parenterally with the estrogenic agent of the present invention as it is or in combination with a pharmacologically acceptable carrier.

  The pharmaceutical preparation of the present invention can be administered orally, for example, as tablets (including sugar-coated tablets and film-coated tablets), pills, granules, powders, capsules (including soft capsules and microcapsules), and syrups. Examples of dosage forms for parenteral administration include injections, infusions, drops, suppositories, and the like. It is also effective to make a sustained-release preparation by combining with an appropriate base.

  As a method for producing the pharmaceutical product of the present invention into the above-mentioned dosage form, a known production method generally used in this field can be applied. In addition, when manufacturing into the above-mentioned dosage forms, excipients, binders, disintegrants, lubricants, sweeteners, interfaces commonly used in the pharmaceutical field when manufacturing into the dosage forms, if necessary. An activator, a suspending agent, an emulsifier, etc. can be appropriately contained and produced.

  For example, when the pharmaceutical product of the present invention is produced into tablets, it can be produced by containing excipients, binders, disintegrants, lubricants, etc., and when produced into pills and granules. , Excipients, binders, disintegrants and the like. In addition, excipients and the like are used for powders and capsules, sweeteners and the like are used for syrups, and suspending agents and surfactants are used for emulsions and suspensions. It can be produced by adding an emulsifier and the like.

  Examples of excipients include lactose, sucrose, glucose, starch, sucrose, microcrystalline cellulose, licorice powder, mannitol, sodium bicarbonate, calcium phosphate, calcium sulfate and the like.

  Examples of the binder include 5 to 10% by weight starch paste solution, 10 to 20% by weight gum arabic solution or gelatin solution, 1 to 5% by weight tragacanth solution, carboxymethyl cellulose solution, sodium alginate solution, glycerin and the like.

  Examples of disintegrants include starch and calcium carbonate.

  Examples of the lubricant include magnesium stearate, stearic acid, calcium stearate, purified talc and the like.

  Examples of sweeteners include glucose, fructose, invert sugar, sorbitol, xylitol, glycerin, simple syrup and the like.

  Examples of the surfactant include sodium lauryl sulfate, polysorbate 80, sorbitan monofatty acid ester, polyoxyl 40 stearate and the like.

  Examples of the suspending agent include gum arabic, sodium alginate, sodium carboxymethyl cellulose, methyl cellulose, bentonite and the like.

  Examples of emulsifiers include gum arabic, tragacanth, gelatin, polysorbate 80 and the like.

  Furthermore, when the pharmaceutical product of the present invention is produced in the above-mentioned dosage form, an appropriate amount of a coloring agent, preservative, fragrance, flavoring agent, stabilizer, thickener, etc., which are usually used in the pharmaceutical field, are added as desired. can do.

  The dose when the pharmaceutical product of the present invention is orally administered varies depending on the sex, symptom, age, and administration method of the patient to be administered, but usually the dose of the compound of the present invention per day for an adult (body weight 60 kg) 1 mg to 500 mg, preferably 1 mg to 70 mg. The daily dose can be administered at once or in several divided doses. It does not matter before meals, after meals, and between meals. The administration period is not particularly limited.

  When administering the pharmaceutical of this invention parenterally, it administers normally in the form of a liquid agent (for example, injection). The single dose varies depending on the administration subject, target organ, symptom, administration method, and the like, but is usually about 1 μg to about 1 mg, preferably about 1 μg to about 200 μg, more preferably about 1 μg to 1 kg body weight in the form of injection, for example. Conveniently, about 1 μg to about 100 μg is administered by intravenous injection. In addition to intravenous injections, subcutaneous injections, intradermal injections, intramuscular injections, intravenous infusions, and the like are included as injections, and sustained-release preparations include iontophoretic transdermal agents and the like. Such an injection is prepared by a method known per se, that is, by dissolving, suspending or emulsifying the compound of the present invention in a sterile aqueous or oily liquid. Aqueous solutions for injection include isotonic solutions (eg, D-sorbitol, D-mannitol, sodium chloride, etc.) containing physiological saline, glucose and other adjuvants, and suitable solubilizers such as alcohol (For example, ethanol), polyalcohol (for example, propylene glycol, polyethylene glycol), nonionic surfactant (for example, polysorbate 80, HCO-50) and the like may be used in combination. Examples of the oily liquid include sesame oil and soybean oil, which may be used in combination with benzyl benzoate, benzyl alcohol or the like as a solubilizer. Buffers (eg, phosphate buffer, sodium acetate buffer), soothing agents (eg, benzalkonium chloride, procaine hydrochloride, etc.), stabilizers (eg, human serum albumin, polyethylene glycol, etc.), preservatives (For example, benzyl alcohol, phenol, etc.) may be blended.

When the pharmaceutical product of the present invention is administered (applied) as a transdermally absorbable external preparation, known additives and the like are mixed as necessary, and creams, solutions, lotions, emulsions, tinctures, and the like. And ointments, aqueous gels, oily gels, aerosols, powders, shampoos, soaps, and other external preparations.
The compounds of the present invention can be made into cosmetics or quasi-drugs as they are or by adding appropriate additives.

  In addition to the above components, the above external preparation includes a water-soluble component, an oily component, a powder component, a surfactant, a polymer component, a thickener, a tackifier, a film forming agent, a pH adjuster, an antioxidant, Preservatives, preservatives, excipients, moisturizers, skin protectants, refreshing agents, fragrances, coloring agents, chelating agents, lubricants, antipruritic agents, blood circulation promoters, astringents, tissue repair promoters, antiperspirants In addition, additives necessary for plant extract ingredients, animal extract ingredients, cosmetics, quasi drugs, and the like can be blended as necessary.

  When administered (applied) as the above-mentioned external preparation, the dose of the compound of the present invention, which is an active ingredient per day for an adult, is usually different depending on the sex, symptoms, administration site, purpose, age, administration method, etc. It is about 1 μg / kg to 10 mg / kg / day, preferably about 1 μg / kg to 1 mg / kg / day. The number of times of application is not particularly limited, but is usually 1 to 5 times a day, preferably 3 to 5 times a day.

  The pharmaceutical of the present invention can be used in combination with drugs such as estrogen preparation, luteinizing hormone preparation, prostatic hypertrophy treatment agent, male hormone preparation, osteoporosis treatment agent, antihyperlipidemic agent, anti-arteriosclerosis agent and the like.

  A food containing the estrogenic agent of the present invention is also encompassed by the present invention (hereinafter also referred to as the food of the present invention). The food of the present invention may be a food consisting only of ingredients derived from Suruga vegetables.

  The food in the present invention may be any general food form containing the compound of the present invention. For example, it can be made into a drink, for example, a soft drink or a powdered drink, by itself or with an appropriate flavor. Specifically, it can be mixed with juice, milk, confectionery, jelly, etc. to eat and drink. It is also possible to provide such food as a health functional food. This health functional food includes foods and drinks that are labeled for use in the present invention such as improvement of symptoms caused by estrogen reduction, particularly food for specified health use, nutritional functional food, and the like.

  Furthermore, the food in the present invention can be used as a food supplement. When used as a food supplement, it can be prepared in the form of tablets, capsules, powders, granules, suspensions, chewables, syrups and the like. The food supplement in the present invention refers to those ingested for the purpose of supplementing nutrition in addition to those ingested as food, and also includes nutritional supplements, dietary supplements and the like.

When the compound of the present invention is ingested as a food, the intake amount of the compound of the present invention per day for an adult (body weight 60 kg) is usually about 1 mg to 300 mg, preferably about 1 mg to 30 mg.
It is preferable to take the above amount in one to several times a day. In this case, the daily intake or the intake per time can be made into one unit packaging.

  EXAMPLES Hereinafter, although a reference example, an Example, and a test example are given and this invention is demonstrated further in detail, an Example is described for description of this invention and does not limit this invention.

(Reagent / Equipment)
Methanol (abbreviated as Wako Pure Chemical Industries, MeOH), Ethanol (abbreviated as Wako Pure Chemical Industries, EtOH), Chloroform (abbreviated as Wako Pure Chemical Industries, CHCl ₃ ), Hexane (abbreviated as Wako Pure Chemical Industries, Hexane), Ethyl acetate (Wako Pure Chemical Industries, Ltd.) Industrial abbreviation EtOAc), acetonitrile (abbreviated as Wako Pure Chemical Industries CH ₃ CN), deuterium methanol (abbreviated as Nippon Oxygen CD ₃ OD), estradiol (abbreviated as Wako Pure Chemical Industries E2), FCS-containing steroid-free DMEM (DULBECCO'S MODIFIED EAGLE'S MEDIUM Modified without phenol red) and Promega's Steady-Glo (registered trademark) Luciferase Assay System was used as a substrate. Caffeic acid (Tokyo Kasei Kogyo), trimethylsilyldiazomethane (Tokyo Kasei Kogyo), and column chromatography used Wakogel 50C18 manufactured by Wako. Thin layer chromatography (abbreviated as TLC) plates were Merck Silica gel 60F254, and ODS plates were Merck RP-18F254s. Vanillin sulfate (vanillin 10 mg, sulfuric acid 1 mL), 13C- and 1H-nuclear magnetic resonance (abbreviated as NMR) were measured with a JEOL GSX-500 nuclear magnetic resonance apparatus. Luminescencer-JNR II AB-2300 ATTA was used as a luminometer. The absorbance meter used was a JASCO V-630 BIO spectrophotometer. A high-performance liquid chromatograph mass spectrometer (abbreviated as LCMS) was SIMADZU LIQUID CHROMATOGRAPH MASS SPECTROMETER LCMS-IT-TOF. A gas chromatograph mass spectrometer (abbreviated as GCMS) was SIMADZU GC-2010 GCMS-QP 2010 plus.

(Reference Example 1)
<Preparation of active ingredients in Suruga vegetable extract>
1. Extraction / Purification (1) Methanol extraction Suruga green leaves (3125 g) were extracted and concentrated with 8.2 L of methanol to obtain a methanol extract (83 g).

(2) Solvent fractionation As shown in the scheme of FIG. 1, the methanol extract of Suruga greens was sequentially fractionated with hexane and ethyl acetate to give a hexane soluble fraction (Hexane ext) and an ethyl acetate soluble fraction (EtOAc ext), a water-soluble fraction (H ₂ O ext) was obtained.

(3) Estrogen assay The cells used were human embryonic kidney 293 (HEK293) cells in which the luciferase gene and estrogen receptors α and β were incorporated. The medium used was steroid-free DMEM containing 5% FCS.
Medium was added to the cells so that the final concentration of the sample was 1 / 100th of the stock concentration. After culturing at 37 ° C. for 24 hours, the substrate was added according to Steady-Glo (registered trademark) Luciferase Assay System, and the luminescence value was measured with a luminometer. The positive control was E2 (estradiol), the negative control was EtOH, and the sample was 4-fold diluted 4 times.
An estrogen assay was performed on each obtained fraction. From this result, it was found that the ethyl acetate soluble fraction showed the strongest estrogenic activity.

2. Search for estrogenic active component from ethyl acetate soluble fraction (1) Column chromatography The ethyl acetate soluble fraction (200 mg) which showed the strongest estrogenic activity was fractionated on a column packed with about 10 g of ODS. The solvent was MeOH—H ₂ O (MeOH: 50 → 60 → 70 → 80 → 90 → 100%).

(2) TLC analysis of fractions Silica gel TLC analysis was performed on the fractions obtained by column chromatography.

(3) Estrogen assay The obtained fraction was subjected to an estrogen assay.

3. Mass fractionation of active ingredients (1) Column chromatography 2. In (3), the estrogenic activity was observed in the fraction obtained at around 50% methanol concentration. Column chromatography was repeatedly performed to collect a large amount of the active ingredient contained in the ethyl acetate soluble fraction. The ethyl acetate soluble fraction (400 mg each) was fractionated on a column packed with about 10 g of ODS. The solvent was MeOH: H ₂ O = 1: 1, and this operation was performed 5 times.

(2) TLC check of fractions TLC check was performed on four fractions A to D obtained by column chromatography.

(3) Estrogen assay The estrogen assay was performed in the above fractions. From this result, it was found that fraction C [B112-6-3-2o frC] has activity (FIGS. 2 and 3).

4). Search for estrogen-like active ingredients from [B112-6-3-2o frC] (1) Column chromatography [B112-6-3-2o frC] (70 mg) with estrogenic activity was packed with about 10 g of ODS. Fractionated on a column. The solvent was MeOH—H ₂ O (MeOH: 20 → 25 → 30 → 35 → 40 → 45 → 50%).

(2) TLC check of fractions TLC check was performed on the fractions obtained by column chromatography.

(3) Estrogen assay An estrogen assay was performed on the fraction obtained by column chromatography. As a result, activity was confirmed in fraction C-2. Therefore, structural analysis of fraction C-2 was performed.

5. Structural analysis of the estrogenic active ingredient C-2 (1) NMR · LCMS measurement active ingredient C-2 results LCMS measurement, the m / z 193 [MH] ^- since the ion peak was observed, the active ingredient C- The molecular weight of 2 was found to be 194.
Subsequently, the active ingredient C-2 was dissolved with CD ₃ OD and measured using one-dimensional NMR. Data were recorded at 500 MHz ⁽¹ H) and ^{125 MHz (13 C) (27} ℃), multiplicity was determined by DEPT method.
The spectral data of ¹ H and ¹³ C NMR of active ingredient C-2 are as shown below.

  From Table 1, methyl caffeate was estimated as the structure of the active ingredient C-2.

Therefore, caffeic acid was methylated using trimethylsilyldiazomethane.
Caffeic acid (5 mg) was taken in an eggplant flask, dissolved in methanol (2 mL), and 10% trimethylsilyldiazomethane in hexane (2 mL) was added until the solution turned yellow. Thereafter, the solvent was removed with an evaporator, and purification was performed with an ODS column. This was used as a sample of methyl caffeate.

  When the sample was subjected to NMR measurement under the same conditions as described above, it was confirmed that it coincided with the NMR spectrum of the active component C-2. Therefore, it became clear that the active ingredient C-2 was methyl caffeate.

(2) Calculation of minimum effective concentration of methyl caffeate In order to obtain the minimum effective concentration of methyl caffeate, an estrogen assay was performed using a methyl caffeate standard. FIG. 5 shows the activity in β cells. This result revealed that the minimum effective concentration of methyl caffeate in the estrogen assay was 6.25 ppm (0.0322 mM).

(Example 1)
Caffeic acid (15 mg) was dissolved in methanol (2 mL) and 10% trimethylsilyldiazomethane in hexane (2 mL) was added until the solution turned yellow. Thereafter, the solvent was removed, and the residue was purified by column chromatography (eluent: ethyl acetate-hexane, 1: 1) to obtain methyl caffeate (10 mg).

(Example 2)
Caffeic acid (50 mg) was dissolved in ethanol (2 mL), Dowex 50 (H ⁺ form) (50 mg) was added, and the mixture was stirred at 60 ° C. in the dark for 3 days. Thereafter, the reaction solution was filtered, the solvent was removed, and the residue was purified by column chromatography (eluent; ethyl acetate-hexane, 1: 1) to obtain ethyl caffeate (20 mg).

Example 3
Caffeic acid (50 mg) was dissolved in 1-propanol (2 mL), Dowex 50 (H ⁺ form) (50 mg) was added, and the mixture was stirred at 60 ° C. in the dark for 3 days. Thereafter, the reaction solution was filtered, the solvent was removed, and the residue was purified by column chromatography (eluent; ethyl acetate-hexane, 1: 1) to obtain propyl caffeate (25 mg).

Example 4
Caffeic acid (50 mg) was dissolved in 1-butanol (2 mL), Dowex 50 (H ⁺ form) (50 mg) was added, and the mixture was stirred at 60 ° C. in the dark for 3 days. Thereafter, the reaction solution was filtered, the solvent was removed, and the residue was purified by column chromatography (eluent; ethyl acetate-hexane, 1: 1) to obtain butyl caffeate (30 mg).

(Test Example 1)
The cells used were human embryonic kidney 293 (HEK293) cells in which a luciferase gene and estrogen receptors α and β were incorporated. The medium used was steroid-free DMEM containing 5% FCS.
Medium was added to the cells so that the final concentration of the caffeic acid ester was 1/100 of the stock solution concentration. After culturing at 37 ° C. for 24 hours, the substrate was added according to Steady-Glo (registered trademark) Luciferase Assay System, and the luminescence value was measured with a luminometer. The positive control was E2 (estradiol), the negative control was EtOH, and the caffeic acid esters of Examples 1 to 4 were diluted four times in four stages.
The horizontal axis shows the relative luminescence intensity when the luminescence intensity when the activity test is performed by adding only the solvent (ethanol) is 1 (negative control). Moreover, the significant difference test with respect to the negative control was performed.

  As shown in FIGS. 4 and 5, all caffeic acid esters showed estrogen activity specific to β cells. The minimum effective concentration was 6.25 ppm for all caffeic acid esters.

Claims (10)

Formula (I):
[Wherein Ar is a C6-C14 aryl group optionally substituted with 1 to 3 substituents selected from the group consisting of a halogen atom, a hydroxy group and a C1-C6 alkoxy group, and A is substituted. An optionally substituted C1-C4 alkylene group, an optionally substituted C2-C4 alkenylene group or an optionally substituted C2-C4 alkynylene group, R ¹ is a hydrogen atom, an optionally substituted C1-C6 An alkyl group, an optionally substituted C3-C6 cycloalkyl group or an optionally substituted C6-C14 aryl group], or an estrogenic agent containing the salt thereof. Ar is phenyl optionally substituted by 1 to 3 substituents selected from the group consisting of a halogen atom, a hydroxy group and a C1-C6 alkoxy group, A is a C2-C4 alkenylene group, and R ¹ is hydrogen The estrogen-like agent according to claim 1, which is an atom or a C1-C6 alkyl group.   The estrogen according to claim 1, wherein the compound represented by the formula (I) or a salt thereof is at least one selected from the group consisting of methyl caffeate, ethyl caffeate, propyl caffeate and butyl caffeate. Like agent.   The estrogenic agent according to any one of claims 1 to 3, wherein the compound represented by the formula (I) or a salt thereof is derived from Suruga greens.   The estrogenic agent according to any one of claims 1 to 4, which is used for treatment and / or prevention of osteoporosis or gynecological diseases.   The estrogenic agent according to any one of claims 1 to 4, which is used for treatment and / or prevention of prostate cancer or benign prostatic hyperplasia.   A pharmaceutical comprising the estrogenic agent according to any one of claims 1 to 6.   The foodstuff containing the estrogenic agent of any one of Claims 1-6.   The food according to claim 8, wherein the food is a health functional food or a dietary supplement.   The food according to claim 9, wherein the health functional food is a food for specified health use or a nutritional functional food.