JPH11246413A - Steroid 5-alpha-reductase ii-type-specific inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject inhibitor having excellent steroid 5α-reductaseII- type-specific inhibitory effect, and useful in e.g. hair growth stimulants, anti- acnevulgaris bacteria skin drugs, by including a triterpene compound as active ingredient. SOLUTION: This inhibitor is obtained by including a compound of formula I (R is a 1-6C alkyl) (e.g. a compound of formula II) as active ingredient which is afforded by separation from extracts of plants such as cuachalalate; juliania adstringens, Amphiptery gumadstringens or the like, belonging to the family Julianiaceae. When this inhibitor is to be used in e.g. medicaments/quasi-drugs and cosmetic compositions, it is preferable that its daily dose is 4-10 ml and the above-mentioned extract (s) is included at 0.005-20 wt.% on a dry basis.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a steroid 5α-reductase type II-specific inhibitor containing triterpenes. INDUSTRIAL APPLICABILITY The inhibitor of the present invention can be effectively used as a hair restorer, a therapeutic agent for anti-acne and the like in the fields of pharmaceuticals, quasi-drugs, cosmetics and the like.

[0002]

Conventionally, male pattern baldness, alopecia, seborrhea and acne (acne vulgaris) are caused by male hormones and metabolites from male hormones. In particular, recent studies have revealed that testosterone, a major male hormone, is metabolized by 5α-reductase into 5α-dihydroxytestosterone (DHT), which causes various skin disorders.

[0003] For this reason, female hormonal agents such as estradiol and anti-androgenic agents have been used in conventional hair restorers.

[0004] Recently, the search for a substance that inhibits the activity of 5α-reductase, which controls the metabolism of testosterone to DHT, has been advanced.

[0005] In male pattern baldness, it has been found that the depilated crown has higher 5α-reductase activity than the undepilated temporal region, and DHT binds to nuclear receptors in cells. While promoting the growth of sebaceous glands, it also acts on hair mother cells to suppress cell growth and prevent hair growth.

[0006] Recent studies have revealed that human 5α-reductase has a type I isozyme that is present in all tissues and a type II isozyme that is also present in androgen target tissues such as the prostate. Was.

[0007] In human scalp hair follicles, both type I and type II isozymes are present, and both are thought to be involved in the progress of male pattern hair loss.

Recently, the search for substances that specifically inhibit 5α-reductase type I and type II has been advanced.

[0009]

However, in the above-mentioned prior art, hormonal agents must be used under the control of a physician from the viewpoint of side effects and safety, and cosmetics for general consumers have restrictions on the blending standards. There are limits to their use in cosmetics.

On the other hand, 5α-reductase activity inhibitors have been found in various plant extracts and have been reported (JP-A-60-146829 and JP-A-63-8).
No. 8121, No. 64-3125, Tokiko 6-4755
No. 4, etc.), however, none of these are inadequate in terms of their action and effect, or have not reached a level where they can be sufficiently satisfied in terms of safety and cost. Although various studies have been made on the active ingredients of these substances,
Many things were not identified.

[0011]

Means for Solving the Problems The present inventors have made intensive studies to solve the above-mentioned problems, and as a result, from plant extracts, 5α-reductase, which is extremely excellent, is specifically superior to type II. Successfully identified and identified a component having an inhibitory activity.

That is, the present invention provides the following general formula (I)

[0013]

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(Wherein R represents an alkyl group having 1 to 6 carbon atoms) or a steroid 5α-containing a pharmaceutically acceptable salt thereof as an active ingredient.
It relates to a reductase type II specific inhibitor.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail.

The steroid 5α-reductase I of the present invention
The type I-specific inhibitor has the following general formula (I):

[0017]

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(Wherein R represents an alkyl group having 1 to 6 carbon atoms) or a pharmaceutically acceptable salt thereof as an active ingredient.

In the above, the alkyl group having 1 to 6 carbon atoms may have any of a straight-chain structure and a branched structure, and specifically includes a methyl group, an ethyl group, an n-propyl group, an isopropyl group and a butyl group. , T-butyl group, pentyl group, hexyl group and the like. In the present invention, a methyl group is particularly preferred.

As used herein, “pharmaceutically acceptable salts” include, for example, alkali metal salts such as sodium and potassium salts; alkaline earth metal salts such as calcium salts,
Magnesium salts and aluminum salts; various organic amines, i.e. amine salts formed from aliphatic or arylaliphatic primary, secondary or tertiary mono-, di- or polyamines or heterocyclic bases, such as triethylamine,
2-hydroxyethylamine, di- (2-hydroxyethyl) amine, tri- (2-hydroxyethyl) amine, 4-aminobenzoic acid-2-diethylaminoethyl ester, 1-ethylpiperidine, bicyclohexylamine, N, N ' -Dibenzyl-ethylenediamine, pyridine, collidine, quinoline, procaine, dibenzylamine, 1-ephenamine and salts derived from N-alkyl-piperidine, but are not limited thereto.

The triterpenes represented by the above general formula (I) have been previously filed by the present applicant as a novel compound which has been separated from a plant extract and successfully identified (Japanese Patent Application No. 9-1990). 62394). Plants that can be used for extraction include cuachalalate (Julianiaceae) of the family Julianiaaceae;
s), Amphiptery gumadstringens, Orthopterygium hua
ncuy and the like, among which quacharate is particularly preferably used. For the extraction, bark of these plants is preferably used, but not limited thereto.

It has been already clarified that the extract of quacharate has high safety and exhibits excellent hair-growth and hair-growth effects (Japanese Patent Laid-Open No. 6-24942). As described above, the present inventors have found a novel active ingredient having an extremely excellent inhibitory effect on 5α-reductase activity from these extracts, and succeeded for the first time in the separation and identification thereof (Japanese Patent Application No. 9-62394). Specification) did not find a type-specific activity inhibitory effect of 5α-reductase.

According to the present invention, a novel active ingredient having an excellent inhibitory activity specifically on type II of steroid 5α-reductase has been found from the above extracts,
The identification was successful. The specific method and the like will be described in detail in Examples.

Since the inhibitor of the present invention has an excellent inhibitory effect on the activity of steroid 5α-reductase type II specifically, it is particularly useful for the prevention and treatment of symptoms related to the activity of steroid 5α-reductase type II. It can be particularly suitably used for various pharmaceutical preparations and cosmetics in the field of medicine and cosmetics, and specifically, for example, cosmetics for prevention and treatment of acne vulgaris, and comprehensive prevention and treatment of acne It can be used for pharmaceutical preparations, skin cosmetics, cosmetics for hair growth and hair growth, etc. Here, the term “hair growth” is used in a concept that includes the promotion of hair growth, prevention of hair loss, and further, the effects of suppressing dandruff and itch.

The inhibitor of the present invention can be used for the prevention or treatment of injuries and diseases caused by the action of steroid 5α-reductase other than those described above. You may mix | blend a diluent, an excipient, etc. according to the objective. The dosage form is also arbitrary, and may be any of oral and parenteral dosage forms. For example, it can be a liquid, syrup-like or tablet dosage form. In particular, for a liquid preparation, physiological saline, ethanol, 1,3-
Butylene glycol can be used as a diluent or carrier. For preparing a creamy preparation, higher fatty acids, higher alcohols, esters and the like can be used as carriers.

The triterpenes represented by the above general formula (I) of the present invention also include isomers thereof within the scope of the present invention.

When the inhibitor of the present invention is used for hair growth, in addition to alkoxycarbonylpyridine N-oxide, vasodilators such as carpronium chloride, assembly extract and acetylcholine derivatives, amino acids such as serine and methionine, vitamin B ₆ , vitamins E and its derivatives, vitamins such as biotin, pantothenic acid and its derivatives, glycyrrhetinic acid and its derivatives, nicotinic acid esters such as benzyl nicotinate, skin function enhancers such as cepharanthin, and female hormones such as estradiol at the same time. You may mix. Further, additives usually used for hair restoration, for example, hinokitiol, hexachlorophene,
Antibacterial agents such as benzalkonium chloride, cetylpyridinium chloride, undecylenic acid, trichlorocarbanilide and bithionol; cooling agents such as menthol; salicylic acid, zinc and its derivatives; drugs such as lactic acid and its alkyl esters; and organic acids such as citric acid. Acids, protease inhibitors such as tranexamic acid, amino acids such as arginine, olive oil, squalane, liquid paraffin, isopropyl myristate, oils such as higher fatty acids and higher alcohols, polyhydric alcohols such as glycerin and propylene glycol, and other interfaces Activators, fragrances, antioxidants, ultraviolet absorbers, dyes, ethanol, water, humectants, thickeners, etc.
It can be appropriately compounded within a range that does not impair the effects of the present invention.

When the inhibitor of the present invention is used as a composition, particularly as a pharmaceutical / quasi-drug or cosmetic composition, its dosage can be varied depending on the form of the composition or the purpose of use, and so is specified. But generally between 4 and 1 per day
It is about 0 ml. Specifically, for example, when the compound of the present invention is obtained from an extract obtained according to the method described in Examples described below, the extract (dry weight) is generally 0.005 to 20% by weight, preferably in pharmaceuticals and cosmetics. Is 0.001
It is blended to be 10% by weight. 0.005% by weight
If it is less than 20%, it is difficult to sufficiently exert the effects of the present invention, while if it exceeds 20% by weight, it is not preferable in terms of formulation.

[0029]

Next, the present invention will be described more specifically with reference to examples. The present invention is not limited by this. The compounding amount is% by weight.

Example 1 Isolation and Purification of the Compound of the Present Invention 2 kg of a dried product of Quacharate was added to a room temperature (about 23 ° C.)
Extraction by immersion in 2 liters of ethanol for 24 hours
Times to obtain an extract, and then the extract is dried under reduced pressure,
40 g of an ethanol extract were obtained.

For 40 g of this ethanol extract, 2 kg of "Diaion HP-20" (manufactured by Nippon Nishui) was used.
The ratio of methanol to distilled water was changed from 40% to 100% to elute. The 100% methanol eluted fraction was collected and dried under reduced pressure to obtain 10 g of the dried product under reduced pressure.

10 g of the dried product under reduced pressure was fractionated using 1 kg of silica gel ("Wakogel C-200") while changing the ratio of hexane to ethyl acetate from 15: 1 to 1: 1. Each fraction was dried under reduced pressure, and the inhibitory activity of 5α-reductase by type was measured by the following method. As shown in Table 1 below, fractions having high inhibitory activity (in Table 1, sample (6))
The product was purified by silica gel column treatment and recrystallization according to a known method to obtain 40 mg of a compound.

With respect to this compound, the following ¹ H-NMR, ¹³
When its structure was determined from the physical property values of C-NMR, the compound 6-oxo-3α-hydroxyeupha-7,
It was found to be 24-dien-26-oic acid.

[0034]

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This compound has been previously filed by the present applicant as a novel compound (Japanese Patent Application No. 9-62394). However, when this compound is 5α
-It has not been reported at all that reductase type II has an activity inhibitory effect specifically.

It should be noted that 5α- of the compound represented by the above formula (4)
The reductase type-based inhibitory activity measurement and tricogram test were performed by the following methods.

I. Measurement of inhibitory activity of each type of 5α-reductase The inhibitory activity of each type of 5α-reductase can be measured by the following method.

That is, for the measurement of 5α-reductase type I-specific inhibitory activity, normal human liver cells (Chang Liver) or dermal papilla cells, which mainly produce type I, are preferably used, and 5α-reductase is preferably used. For measurement of the type II-specific inhibitory activity, human neonatal foreskin gland fibroblasts (Hs-68) and prostate cells, which mainly produce type II, are preferably used. In this example, normal human liver cells (Chang Liver) were used for type I measurement, and live human neonatal foreskin gland fibroblasts (Hs-68) were used for type II measurement.

Using each of these human-derived cells, the amount of testosterone reduced to DHT or the amount of androstenedione reduced to androstanedione was measured in the cell culture supernatant. The steroid 5-α-reductase inhibitory activity of a 70% ethanol solution of the compound represented by Chemical Formula 4 obtained by separation and purification can be measured by type, and the inhibition rate can be determined according to the following formula. In this example, the measurement was performed by measuring the amount of androstenedione reduced to androstanedione. Table 1 shows the results.

In Table 1, sample (1) is the ethanol extract described in Example 1;
(4) each elution fraction at the end of the operation by "Diaion HP-20" column chromatography;
Samples (5) and (6) are elution fractions using sample (4); sample (7) was isolated and purified from sample (6). That is, from the results in Table 1, the highly polar fraction of the fraction 2 eluted with hexane: ethyl acetate (1: 1) had a high inhibitory activity, and the fractions were collected, treated with a silica gel column and recrystallized to form 6-ethyl acetate. oxo-3α-hydroxyeupha-7,24-dien-26
40 mg of -oic acid were obtained.

[0041]

(Equation 1)

[0042]

[Table 1]

II. Tricogram Test The effect of the present invention on hair restoration (that is, promotion of hair growth or prevention of hair loss) can be evaluated by performing the following tricogram test. (Preparation of test sample) The 6-oxo-3α-hydroxyeupha-7,24-dien- obtained in Example 1 was added to 70% by weight of 95% ethanol.
0.1% by weight of 26-oic acid and 0.5% by weight of an adduct of hydrogenated castor oil with ethylene oxide (40 mol) were added and dissolved by stirring. Next, ion-exchanged water was added to 100% by weight and mixed to prepare a liquid hair nourishing material. As a control, 6-oxo-3α-hydroxyeupha-7,24-dien-
A composition containing no 26-oic acid (control) was prepared. (Evaluation by tricogram test) A tricogram test was performed by a male panel (18 persons) using the test samples (product of the present invention and control product).

That is, the hair roots of the extracted hair before and after the use of the test sample are observed under a microscope, the number of hair roots at the stasis stage is counted from the form of the hair root, and the hair growth effect of the hair nourishing agent is compared by increasing or decreasing the ratio. did. A telogen follicle is a hair follicle that has stopped growing, and it has been recognized that the proportion of this telogen follicle is higher in people who report hair loss than in normal people.

Specifically, the product of the present invention was applied to the scalp of 10 male panels and the control product was applied to the scalp of 8 other male panels twice a day, 2 ml at a time, for 6 months continuously. Immediately before application and immediately after application for 6 months, 100 hairs were removed from each subject, and the percentage of resting roots before and after application was examined. Of the hair follicles) was measured and measured. Table 2 shows the results.

[0046]

[Table 2]

Example 2 Test for Improvement of Acne Vulgaris (Acne Symptoms) A lotion having the following composition was prepared as a test sample. In addition, the compounding amount is weight%. (Phase A) Sorbitol 3.0 Glycerin 5.0 Resorcin 0.02 Ion-exchanged water Residue (Phase B) 6-oxo-3α-hydroxyeupha-7,24-dien-26-oic acid (Example 1) 1 Polyoxyethylene (60) hydrogenated castor oil 0.5 95% ethanol 20.5 Perfume Appropriate amount (Preparation method) Mix and dissolve each component of phase A, and add the mixed solution of phase B to this while stirring. A lotion was prepared as a homogeneous solution.

As a control, 6-oxo-3α-hydroxyeupha-7,24-dien-26-oic acid was used as a component of the above lotion.
Was prepared without the addition of

Using the test samples (product of the present invention and control product), 40 males and 40 females ages 13 to 23 were used as panels.
After one week, the effect of improving acne symptoms was determined according to the following evaluation criteria. Table 3 shows the results. (Evaluation criteria) Complete cure: No symptoms at all Significant effect: Significant improvement effect was observed Effective: Symptoms were lightened overall Invalid: No change in symptoms before and after use

[0050]

[Table 3]

Example 3 Cream (Phase A) Beeswax 10.0 Paraffin Wax 6.0 Lanolin 3.0 Isopropyl Myristate 6.0 Squalane 8.0 Liquid Paraffin 26.0 Polyoxyethylene Sorbitan Stearate 2.0 Sorbitan Monostearate 4.2 Preservative Appropriate amount (phase B) Propylene glycol 2.0 Polyoxyethylene (60) hydrogenated castor oil 1.0 6-oxo-3α-hydroxyeupha-7,24-dien-26-oic acid 0 1.1 Purified water residue (Preparation method) Mix the ingredients of phase A, heat and dissolve at about 75 ° C, add the combined liquid of phase B heated to 75 ° C with stirring, and then heat to 45 ° C. Continue stirring while cooling to
The cream was obtained by standing.

When this cream was subjected to a practical use test on humans in the same manner as in Example 2, it was confirmed that the cream was excellent in improving acne symptoms.

Example 4: O / W emulsion type hair restorer (A phase) 6-oxo-3α-hydroxyeupha-7,24-dien-26-oic acid 0.01 Polyoxyethylene (60 mol) addition-hardened castor oil 2.0 Glycerin 10.0 Dipropylene glycol 10.0 1,3-butylene glycol 5.0 Polyethylene glycol (molecular weight 1500) 5.0 (Phase B) Cetyl isooctanoate 10.0 Squalane 5.0 Vaseline 2.0 Propylparaben 2.0 (C phase) Carboxyvinyl polymer 1% aqueous solution 30.0 Sodium hexametaphosphate 0.03 Ion exchange water 8.35 (D phase) Ion exchange water 4.5 (E phase) Kaseikari 0.12 Ion exchange Water residue (Production method) A phase and B phase are each heated and dissolved at 60 ° C.
The mixture was mixed and treated with a homomixer to obtain a gel substance. Phase D is gradually added thereto, and dispersed by a homomixer. Then, dissolved C phase is added thereto, and further dissolved E phase is added, and emulsified by a homomixer to obtain an O / W emulsion type hair restorer. Was.

When this hair regrowth was subjected to a practical use test on humans in the same manner as in Example 2, it was confirmed that the hair regrowth effect was excellent.

Example 5: Creamy hair restorer (Phase A) 6-oxo-3α-hydroxyeupha-7,24-dien-26-oic acid 1.0 Liquid paraffin 5.0 Cetostearyl alcohol 5.5 Glyceryl monostea Rate 3.0 EO (20 mol) -2-octyldodecyl ether 3.0 Propylparaben 0.3 Fragrance 0.1 (Phase B) Glycerin 8.0 Dipropylene glycol 20.0 Polyethylene glycol (molecular weight 4000) 5.0 Sodium hexametaphosphate 0.005 Ion-exchanged water Residue (Production method) A phase and B phase are each heated and dissolved and mixed.
The mixture was emulsified with a homomixer to obtain a creamy hair restorer. An actual use test was carried out on the human hair by the tricogram test described in Example 1, and it was confirmed that the hair growth effect was excellent.

Example 6 Hair Tonic 6-oxo-3α-hydroxyeupha-7,24-dien-26-oic acid 10.0 Peppermint (1,3-butylene glycol solution) 0.1 N, N-dimethyl-2 -Dodecylamine oxide 1.0 Hinokitiol 1.0 Vitamin B ₆ 0.2 Vitamin E acetate 0.02 Menthol 0.2 Assembly extract 1.0 Salicylic acid 0.1 Mica (ethanol extract) 0.5 Propylene glycol 2.0 Sodium hyaluronate 0.01 Polyoxyethylene (10 mol) monostearate 2.0 75% ethanol Residue

Example 7: Hair tonic 6-oxo-3α-hydroxyeupha-7,24-dien-26-oic acid 10.0 Altea (ethanol extract) 1.5 Yokuinin (ethanol extract) 1.5 N, N-dimethyl-2-tetradecylamine oxide 0.05 Hinokitiol 1.0 Vitamin B ₆ 0.2 Vitamin E acetate 0.02 Menthol 0.2 Salicylic acid 0.1 Caccon (ethanol extract) 0.5 Propylene glycol 0. 01 Sodium hyaluronate 0.01 Polyoxyethylene (10 mol) monostearate 2.0 70% ethanol Residue

Example 8: Hair tonic 6-oxo-3α-hydroxyeupha-7,24-dien-26-oic acid 0.6 95% ethanol 50.0 Monoammonium glycyrrhizinate 0.05 Althea (ethanol extract) 0 .05 Yokuinin (1,3-butylene glycol solution) 0.05 Peppermint (ethanol extract) 0.05 Saffron (ethanol extract) 0.05 Rosemary (ethanol extract) 0.05 Senkyu (ethanol extract) 0 .05 Touki (ethanol extract) 0.05 Diou (ethanol extract) 0.05 Silkworm (ethanol extract) 0.05 Sodium lauryl sulfate 0.1 N, N-dimethyl-2-tridecylamine oxide 0.5 Hydrogenated castor oil ethylene oxide (40 mol) adduct 0.5 succinic acid qs perfume qs pigment Volume Purified water Residue

Example 9: Aerosol hair restorer (stock solution formulation) 6-oxo-3α-hydroxyeupha-7,24-dien-26-oic acid 0.6 95% ethanol 50.0 Glycyrrhetinic acid 0.1 Artair (ethanol extraction Liquid) 0.05 ぺ permint (ethanol extract) 0.05 assembly extract 0.1 sodium lauryl sulfate 0.1 N, N-dihydroxymethyl-2-0.2 decylamine oxide hydrogenated castor oil ethylene oxide (40 mol) addition Product 0.5 Lactic acid proper amount Sodium lactate proper amount Fragrance proper amount Pigment proper amount Purified water residue (filling formula) Stock solution 50.0 Liquefied petroleum gas 50.0 After dissolving the stock solution formulation, fill it in a can and fill it with a valve. After mounting, fill with gas. An actual use test was performed on this hair tonic by the tricogram test described in Example 1 above, and it was confirmed that the hair growth effect was excellent.

[0060]

As described in detail above, the inhibitor of the present invention has a particularly excellent steroid 5α-reductase type II-specific inhibitory effect, and is useful in the fields of pharmaceuticals, quasi-drugs, cosmetics, etc. It can be effectively used as a therapeutic agent for acne and the like.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁶ Identification symbol FI // C07J 9/00 C12N 5 / 00F (72) Inventor Yasuo Fujimoto 8-32-8 Oizumi Gakuen-cho, Nerima-ku, Tokyo

Claims (4)

[Claims] 1. A compound represented by the following general formula (I) (Wherein, R represents an alkyl group having 1 to 6 carbon atoms) or a steroid 5α-reductase type II specific inhibitor comprising, as an active ingredient, a compound represented by the formula: . 2. The steroid 5α-reductase II according to claim 1, wherein in the general formula (I), R is —CH _3.
Type-specific inhibitors. 3. The steroid 5α-reductase type II specific inhibitor according to claim 1, which is used as a hair restorer. 4. The steroid 5α-reductase II according to claim 1, which is used as a therapeutic agent for skin disorders.
Type-specific inhibitors.