JP7614652B2 - Steroid 5α-reductase inhibitors - Google Patents

Description

The present invention relates to a steroid-5α-reductase inhibitor, a hair growth agent, a prostate hyperplasia suppressant, and an agent for preventing and/or improving acne.

One of the causes of male pattern hair loss is known to be the accumulation of large amounts of male hormones in the metabolic system, which increases the action of male hormones. It is believed that testosterone is reduced to dihydrotestosterone (DHT) by 5α-reductase, and the action of the reduced DHT shortens the hair growth phase. Therefore, it is believed that male pattern hair loss caused by increased action of male hormones can be prevented or reduced by inhibiting the reduction of testosterone to DHT, i.e., by inhibiting steroid-5α-reductase (testosterone-5α-reductase) (Non-Patent Document 1).

It has also been established that DHT is the main cause of prostatic hyperplasia, and 5α-reductase inhibitors are known to be effective in treating prostatic hyperplasia (Non-Patent Document 2).

It is known that increased production of DHT increases sebum secretion and leads to the development of seborrheic alopecia and acne (acne vulgaris). Therefore, it is believed that by inhibiting the production of DHT, seborrheic alopecia and acne can be prevented or reduced.

Propolis is a glue-like substance made by mixing tree sap, plant buds and exudates collected by honeybees with beeswax and other substances, and is known to have antibacterial, antioxidant, anti-inflammatory, antitumor and anti-allergic effects.

The components of propolis are influenced by the type and composition of the original plant from which it is made, so it is known that the types and amounts of active ingredients contained in propolis, as well as their physiological activity, can vary greatly depending on the country or region where it is harvested.

In recent years, research has been conducted into the components of propolis from different regions. For example, the main components of Brazilian propolis, which is derived from Baccharis doracunclifolia, are p-coumaric acid, artepillin C, drupanin, baccharin, and other cinnamic acid derivatives. On the other hand, the main components of propolis from China, Europe, and Australia, which is derived from poplar, are flavonoids such as chrysin and galangin.

Patent document 1 reports that extracts of Chinese and Brazilian propolis mixed with hydrophilic solvents or water have excellent effects in inhibiting testosterone-5α-reductase.

Patent Document 2 discloses that nemorosone contained in Cuban propolis has testosterone-5α-reductase inhibitory activity, and therefore nemorosone and Cuban propolis are useful as active ingredients in hair growth agents, prostate hyperplasia inhibitors, and anti-acne agents.

JP 2000-53539 A Patent No. 6300436

AL. Dallob et al. , The Effect of Finasteride, a 5α-Reductase Inhibitor, on Scalp Skin Testosterone and Dihydrotestosterone Concentrations in Patients with Male Pattern Baldness. J Clin Endocrinol Metab, 79, 703-706 (1994) S. Tu-Yue et al. , A Novel In Vitro Model to Screen Steroid 5α-Reductase Inhibitors Against Benign Prostatic Hyperplasia. Meth Find Exp Clin Pharmacol, 20(4), 283-287 (1998)

The present invention aims to provide a steroid-5α-reductase inhibitor, a hair growth agent, a prostate hyperplasia suppressant, and an agent for preventing and/or improving acne.

Since propolis is made from plants harvested by honeybees, its ingredients vary depending on the place of origin, and it is expected that its effectiveness will vary depending on the place of origin. Therefore, the inventors conducted extensive research to achieve the above-mentioned objective, and discovered that propolis derived from plants of the genus Lepidosperma, propolis derived from plants of the genus Macaranga, and propolis derived from plants of the genus Betula have high steroid-5α-reductase inhibitory activity.

The present invention was completed based on these findings and through further investigation, and provides the following steroid 5α-reductase inhibitor, hair growth agent, prostate hyperplasia suppressant, and acne prevention and/or improvement agent.

Item 1. A steroid-5α-reductase inhibitor comprising at least one selected from the group consisting of propolis derived from plants of the genus Lepidosperma, propolis derived from plants of the genus Macaranga, and propolis derived from plants of the genus Betula.
Item 2. A hair growth agent comprising the steroid 5α-reductase inhibitor according to item 1.
Item 3. A prostatic hyperplasia inhibitor comprising the steroid-5α-reductase inhibitor according to item 1.
Item 4. An acne prevention and/or amelioration agent comprising the steroid 5α-reductase inhibitor according to item 1.
Item 5. The agent according to any one of Items 1 to 4, which is a food or drink, a cosmetic, a medicine, or a quasi-drug.

Propolis derived from plants of the genus Lepidosperma, propolis derived from plants of the genus Macaranga, and propolis derived from plants of the genus Betula have excellent steroid-5α-reductase inhibitory activity, and are therefore useful as active ingredients in steroid-5α-reductase inhibitors, hair growth agents, prostate hyperplasia suppressants, and acne prevention and/or improvement agents.

In addition, propolis has traditionally been used as a food ingredient, so it is highly safe.

The following describes in detail the embodiments of the present invention.

In this specification, the word "comprise" includes the meanings "essentially consisting of" and "consisting only of."

The katakana reading of "Reductase" is generally written as "reductase" or "riductase", but these have the same meaning. In the present invention, the katakana reading of "Reductase" is "riductase".

In addition, in the present invention, steroid-5α-reductase and testosterone-5α-reductase are considered to have the same meaning.

The steroid-5α-reductase inhibitor of the present invention is characterized by containing at least one propolis selected from the group consisting of propolis derived from plants of the genus Lepidosperma, propolis derived from plants of the genus Macaranga, and propolis derived from plants of the genus Betula.

Propolis is a resinous or waxy substance that forms the walls of beehives, and is made by mixing the buds and resins of plants that bees have collected with their saliva. The plants from which the propolis of the present invention is derived are plants of the genus Lepidosperma, propolis derived from plants of the genus Macaranga, and plants of the genus Betula.

The propolis used in the present invention is not particularly limited as long as it is derived from the above-mentioned plants, and may be derived from any of the places of origin, such as Brazil, China, Taiwan, European countries, Russia, Oceania, and America, among which Australian propolis, American propolis, Taiwanese propolis, and Russian propolis are preferred. Propolis derived from plants of the Lepidosperma genus includes Australian propolis, propolis derived from plants of the Macaranga genus includes American propolis and Taiwanese propolis, and propolis derived from plants of the Betula genus includes Russian propolis.

The propolis in the present invention includes, for example, raw propolis, its pulverized product, supercritical extract, and extract with water and/or hydrophilic organic solvent. Among them, extract with water and/or hydrophilic organic solvent, supercritical extract, etc. are preferred, and extract with water and/or hydrophilic organic solvent is more preferred. Extract with water and/or hydrophilic organic solvent is preferred because propolis components are extracted efficiently and in a balanced manner in a short time, and large-scale equipment is not required when scaling up.

Examples of hydrophilic organic solvents include lower alcohols such as ethanol, methanol, and propanol, ketones (acetone, methyl ethyl ketone, methyl butyl ketone, etc.), esters (methyl acetate, ethyl acetate, etc.), glycols (ethylene glycol, propylene glycol, etc.), glycerin, tetrahydrofuran, acetonitrile, acetic acid, propionic acid, acetamide, dimethylformamide, and dimethyl sulfoxide. Lower alcohols are preferred, and ethanol is particularly preferred.

These extraction solvents can be used alone or in any combination of two or more to form a mixed solution. Preferred are water, lower alcohol (preferably ethanol) or a mixture thereof (water-containing alcohol, preferably water-containing ethanol). When water-containing alcohol (preferably water-containing ethanol) is used as the extraction solvent, the alcohol concentration is not limited, but is preferably 50 to 100% by volume, and particularly preferably 70 to 100% by volume.

For extraction, raw propolis blocks, raw propolis blocks washed with alcohol, etc. can be used, and the extraction efficiency can be improved by using raw propolis blocks cut or crushed to an appropriate size. The volume of the extraction solvent used is, for example, 1 to 20 times, preferably 2 to 10 times, the mass of the raw propolis blocks.

Extraction can be performed by adding propolis to a solvent, for example, at a temperature between 0°C and the temperature at which the solvent boils (usually a temperature below 100°C), preferably between 2°C and 40°C. Extraction can be performed at the above temperature for a specified period of time (for example, one hour or more) with stirring or by leaving the mixture to stand. After extraction, a process such as filtration can be performed to remove insoluble matter, and the extract can be obtained.

The solvent extract thus obtained can be used as is, or, if necessary, can be further purified to increase the content of active ingredients by ultrafiltration, molecular sieve chromatography (gel filtration), adsorption chromatography, ion exchange chromatography, affinity chromatography, high performance liquid chromatography (HPLC), dialysis, or a combination of these.

The solvent extract can also be concentrated and dried by freeze-drying, spray-drying, or the like. Concentration, freeze-drying, and spray-drying can be carried out according to conventional methods. The propolis extract of the present invention also includes powders obtained by powdering such extracts, and granular forms obtained by granulating the powders.

The steroid-5α-reductase inhibitor of the present invention may contain other known ingredients as appropriate, provided that they do not impair the effects of the present invention.

The content of propolis in the steroid-5α-reductase inhibitor of the present invention is not particularly limited as long as the effects of the present invention are obtained, and can be appropriately adjusted depending on the final form, etc., and can be, for example, 0.00001% by mass or more, 0.0001% by mass or more, 0.001% by mass or more, 0.01% by mass or more, 0.1% by mass or more, 0.5% by mass or more, 1% by mass or more, 3% by mass or more, 5% by mass or more, 7% by mass or more, 10% by mass or more, 15% by mass or more, based on the total solid content of the steroid-5α-reductase inhibitor. The above may be 20% by mass or more, 30% by mass or more, 40% by mass or more, 50% by mass or more, 60% by mass or more, 70% by mass or more, 80% by mass or more, or 90% by mass or more, and 100% by mass or less, 90% by mass or less, 80% by mass or less, 70% by mass or less, It may be 60 mass% or less, 55 mass% or less, 50 mass% or less, 40 mass% or less, 30 mass% or less, 20 mass% or less, 15 mass% or less, 10 mass% or less, 7 mass% or less, 5 mass% or less, 3 mass% or less, 1 mass% or less, or 0.5 mass% or less.

Propolis is preferably applied periodically to inhibit steroid-5α-reductase, and particularly preferably applied once a day or in divided doses. Although not particularly limited, propolis can be used at a dose of 1 μg to 1,000 mg per day for an adult weighing 60 kg, preferably at a dose of 2 μg to 500 mg, more preferably at a dose of 3 μg to 250 mg, and even more preferably at a dose of 4 μg to 100 mg, in solid form, depending on factors such as the health condition of the person to be administered, the method of administration, and the combination with other agents.

The steroid-5α-reductase inhibitor of the present invention can be used as cosmetics, food and drink (particularly food and drink intended for health maintenance or promotion (e.g., health food, functional food, nutritional composition, dietary supplement, supplement, health food, food for specified health use, nutritional functional food, or food with functional claims)), quasi-drugs, medicines, etc. The steroid-5α-reductase inhibitor of the present invention also includes the meaning of an additive that imparts a steroid-5α-reductase inhibitory effect.

In addition to the propolis, the above-mentioned cosmetics can contain ingredients that are normally used in cosmetics, such as disinfectants, preservatives, surfactants, alcohols, aqueous components, water, colorants, pH adjusters, solubilizers, abrasives, foaming agents, enzymes, flavorings, chelating agents, excipients, thickeners, bases, emulsifiers, solvents, stabilizers, oils, cleaning agents (lactic acid bacteria), etc., as needed.

Cosmetics include any cosmetic product that is applied to the skin, mucous membranes, body hair, head hair, scalp, nails, teeth, facial skin, lips, etc. of animals (including humans).

Cosmetics can take a wide range of formulations, including aqueous solutions, solubilized solutions, emulsions, powders, oils, gels, ointments, aerosols, two-layer water-oil systems, and three-layer water-oil-powder systems.

The use of the cosmetics is also arbitrary. For example, basic cosmetics include face wash, lotion, milky lotion, cream, gel, essence, beauty serum, pack, mask, etc.; makeup cosmetics include foundation, lipstick, blusher, eye shadow, eyeliner, mascara, etc.; nail cosmetics include nail polish, base coat, top coat, nail polish remover, etc.; and other products include face wash, (paste or liquid) dentifrice, mouthwash, massage agent, cleansing agent, aftershave lotion, preshave lotion, shaving cream, body soap, soap, shampoo, rinse, hair treatment, hair styling product, hair tonic, hair growth agent, antiperspirant, bath additive, etc.

The above-mentioned propolis can be used as is in the above-mentioned foods and beverages, or, if necessary, can be mixed with minerals, vitamins, flavonoids, quinones, polyphenols, amino acids, nucleic acids, essential fatty acids, cooling agents, binders, sweeteners, disintegrants, lubricants, colorants, flavorings, stabilizers, gelling agents, preservatives, sustained-release regulators, surfactants, solubilizers, wetting agents, etc.

Food and drink includes any food and drink that can be consumed by animals (including humans). The types of food and drink are not particularly limited, and examples include dairy products; fermented foods (yogurt, etc.); beverages (soft drinks such as coffee, juice, and tea drinks, dairy drinks, lactic acid bacteria drinks, drinks containing lactic acid bacteria, yogurt drinks, carbonated drinks, sake, Western liquor, and alcohol such as fruit wine); spreads (custard cream, etc.); pastes (fruit paste, etc.); Western confectionery (chocolate, donuts, pies, cream puffs, gum, gummy candies, jellies, candies, cookies, cakes, puddings, etc.); Japanese confectionery (daifuku, mochi, manju, castella, anmitsu, yokan, etc.); frozen desserts (ice cream, popsicles, sorbets, etc.); foods (curry, beef bowl, porridge, miso soup, soup, meat sauce, pasta, pickles, jam, etc.); and seasonings (dressings, furikake, umami seasonings, soup bases, etc.).

The method for producing the food or beverage is not particularly limited, and any known method can be used as appropriate.

The dosage unit form when used as a supplement is not particularly limited and can be selected appropriately, and examples include chewable tablets, troches, capsules, granules, liquids, powders, syrups, pastes, drinks, gummies, etc.

The above-mentioned propolis can be used alone in the above medicines, or it can be mixed with other medicinal ingredients listed in the Japanese Pharmacopoeia, such as vitamins and herbal medicines.

When the steroid-5α-reductase inhibitor of the present invention is prepared as a pharmaceutical product, the propolis can be prepared together with pharmaceutical acceptable ingredients into the form of tablets (including plain tablets, sugar-coated tablets, effervescent tablets, film-coated tablets, chewable tablets, troches, etc.), capsules, pills, powders (dispersed medicines), fine granules, granules, liquids, suspensions, emulsions, syrups, pastes, injections (including the case where it is mixed with distilled water or infusions such as amino acid infusions or electrolyte infusions at the time of use to prepare a liquid), etc., to form a pharmaceutical preparation.

Medicinal products may be administered locally or systemically. There are no particular limitations on the method of administration, and they may be administered orally or parenterally. Parenteral administration routes include subcutaneous, intradermal, intramuscular, intraperitoneal, intravenous or intraarterial administration, and transdermal administration.

In addition to propolis, the pharmaceutical product of the present invention may contain, as necessary, pharma- ceutically acceptable ingredients such as excipients, binders, disintegrants, lubricants, colorants, suspending agents, thickeners, antioxidants, absorption promoters, pH adjusters, preservatives, stabilizers, surfactants, sweeteners, flavorings, and fragrances.

The cosmetics and pharmaceuticals of the present invention also include quasi-drugs.

The steroid-5α-reductase inhibitor of the present invention described above is applicable to mammals, including humans (preferably humans).

At least one selected from the group consisting of propolis derived from plants of the genus Lepidosperma, propolis derived from plants of the genus Macaranga, and propolis derived from plants of the genus Betula, which the steroid-5α-reductase inhibitor of the present invention contains as an active ingredient, has steroid-5α-reductase inhibitory activity as shown in the examples described below, and is therefore useful as an active ingredient of the steroid-5α-reductase inhibitor. For this reason, the steroid-5α-reductase inhibitor of the present invention can also be used as a hair growth agent, a prostate hypertrophy inhibitor, and an acne prevention and/or improvement agent. The above description regarding the steroid-5α-reductase inhibitor can be applied to the hair growth agent, the prostate hypertrophy inhibitor, and the acne prevention and/or improvement agent. In other words, the content and dosage of propolis in the hair growth agent, the prostate hypertrophy inhibitor, and the acne prevention and/or improvement agent of the present invention are the same as those described above.

The hair growth agent, prostate hyperplasia inhibitor, and acne prevention and/or improvement agent of the present invention can be used as cosmetics, food and drink (particularly food and drink intended for health maintenance and promotion, etc. (e.g., health foods, functional foods, nutritional compositions, dietary supplements, supplements, health foods, foods for specified health uses, nutritional functional foods, or foods with functional claims)), quasi-drugs, medicines, etc. The hair growth agent, prostate hyperplasia inhibitor, and acne prevention and/or improvement agent of the present invention also include the meaning of additives that impart hair growth effects, prostate hyperplasia inhibitor effects, and acne prevention and/or improvement effects. The cosmetics, food and drink, quasi-drugs, and medicines are the same as those described above.

In the present invention, hair growth includes the concepts of hair growth, hair nourishment, and hair loss prevention. The hair growth agent of the present invention may contain other ingredients (finasteride, minoxidil, t-flavanone, Panax ginseng extract, Panax ginseng extract, Swertia japonica extract, 6-benzylaminopurine, adenosine, ethinyl estradiol, forskolin, etc.) to enhance the hair growth effect.

In addition, propolis has traditionally been used as a food ingredient, so it is highly safe.

The present invention will be described in detail below with reference to examples. However, the present invention is not limited to these examples.

Example 1 (Preparation of propolis extract)
The propolis raw mass was pulverized, and 80% by volume ethanol was added and stirred at room temperature for 24 hours to extract the propolis. The extract was filtered through a filter paper (No. 2, manufactured by Advantec Toyo Co., Ltd.) and then concentrated under reduced pressure using an evaporator. The concentrated liquid was freeze-dried to obtain a propolis extract.

Test Example 1 (Method for measuring steroid-5α-reductase inhibitory activity)
Steroid-5α-reductase isolated from Wistar rat liver by conventional centrifugation was used.

Australian propolis extract (derived from plants of the genus Lepidosperma) was diluted with DMSO to final concentrations of 10, 3, 1, and 0.3 μg/ml, American and Taiwanese propolis extracts (derived from plants of the genus Macaranga) were diluted with DMSO to final concentrations of 10, 2, 0.4, and 0.08 μg/ml, and Russian propolis extract (derived from plants of the genus Betula) was diluted with DMSO to final concentrations of 100, 20, 4, 0.8, and 0.16 μg/ml.

Each diluted propolis was mixed with DTT buffer (dithiothreitol buffer) (pH 6.5) containing 20 μg/ml steroid-5α-reductase containing 0.9 μM testosterone and 50 μM NADPH (nicotinamide adenine dinucleotide phosphate), and incubated at 37°C for 30 minutes. The reaction was stopped by adding 1N HCl, and neutralized with 1N NaOH. Testosterone was quantified using a Testosterone EIA Kit. DMSO, a sample diluent, was used as a control, and the inhibition rate was calculated by the following calculation method. The 50% inhibitory activity concentration (IC50: μg/ml) was calculated from the obtained inhibition rate and the active concentration.
Inhibition rate (%)=[(absorbance intensity of sample−absorbance intensity of control)/absorbance intensity of control]×100

The results in Table 1 show that propolis from Australia, the United States, Taiwan and Russia have superior steroid-5α-reductase inhibitory activity to the Brazilian and Chinese propolis disclosed in JP 2000-53539 A and the Cuban propolis disclosed in JP 6300436 A.

Claims (2)

A steroid-5α-reductase inhibitor comprising at least one member selected from the group consisting of propolis derived from plants of the genus Lepidosperma and propolis derived from plants of the genus Macaranga. The agent according to claim 1 , which is a food or beverage, a cosmetic, a medicine or a quasi-drug.