JPH0647554B2 - Testosterone 5α-reductase inhibitor - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION <Industrial field of application> The present invention is applicable to hair growth / hair growth, acne cosmetics and pharmaceuticals, and has no side effects, and testosterone 5α-
It relates to a reductase inhibitor.

<Prior Art> Male baldness, alopecia, seborrhea, acne (acne vulgaris), etc. are said to be due to an excess of male hormones.
In particular, it is known that the main body of this androgen activity in organs such as hair roots and sebaceous glands is 5α-dihydrotestosterone (DHT), which is produced by reducing testosterone by testosterone 5α-reductase (TSR) in these target organs. ing. That is, testosterone (male hormone) biosynthesized and secreted in the testis and adrenal reaches the sebaceous gland by the bloodstream and is converted into DHT by TSR in the sebaceous gland cells. This DHT binds to an intracellular receptor and acts on the nucleus to promote the growth of sebaceous gland cells, while it also acts on hair mother cells to suppress cell division and prevent hair growth. .

Conventionally, for the treatment of male pattern baldness, alopecia or acne,
Substances having female hormone action such as estradiol, substances having anti-androgen action such as cyproterone acetate, substance having strong oxidizing power Chlorine dioxide (ClO)
₂ ) was used. In addition, focusing on the mechanism of action of testosterone described above, research on substances that inhibit TSR activity is also underway.

<Problems to be solved by the invention> However, hormones and oxidants have side effects and safety problems, and cannot be incorporated into cosmetics, or can be used only under the supervision of a doctor. There were only things.

Also, substances reported as substances that inhibit TSR activity have various problems such as insufficient action / effect, insufficient safety, or high cost. To do.

<Means for Solving the Problems> For many years, we have been screening herbal extracts that have no side effects or safety problems and can be incorporated into cosmetics. Then, it has been found that the application of a specific herbal extract is effective for the above problems.

That is, dry powder of medicinal herbs was immersed in 99.5% or 50% ethanol at a ratio of 1 g / 10 ml, and allowed to stand for 1 week and then filtered, and the filtrate was separated to examine the TSR activity. As a result, sage ( Salvia officinalis L.), hop ( Humulus
lupulus L.), rosemary ( Rosmarinus officinalis
L.), St. John's wort (Hypericum erectum Thunb.), Peppermint (Mentha arvensis L.var. Piperascens Malinv. ), Peppermint (Mentha piperita L.), chamomile (Matrica
ria chamomilla L.), Arnica montana L.),
It was found that each ethanol extract of Thyme ( Thymus vulgaris L.) was effective.

<Action> Table 1 shows the TSR activity inhibition rate and protein denaturation rate of the above herbal extracts.

Rat liver homogenate as TSR solution 9,000xg
The supernatant (S-9) was used. First, testosterone 3 μl
Is dissolved in 10 drops of propylene glycol, and 5 ml of Tris-HCl buffer (pH 7.2) is added. To this, NADPH 5mg, S-9
After adding 1 ml and 0.5 ml of the herb extract and incubating at 37 ° C. for 30 minutes, 50 ml of dichloromethane was added to stop the reaction, and the dichloromethane layer was separated and dried under reduced pressure.
Gas chromatography (GC) was used to detect and quantify reaction products such as DHT and androstanediol (G
C condition: column; ULBON HR-1, carrier gas; He 20m
l / min, column temperature; 200 ° C). The TSR activity inhibition rate was determined by comparing the amount of the reaction product when the herb extract was not added (control) with the amount of the reaction product when the herb extract was added.

On the other hand, the protein denaturation rate includes 0.02% ovalbumin.
0.15M sodium sulfate / 0.05M phosphate buffer (pH7.0) 9m
Add 1 ml of the herb extract to 1 l, stir for 5 minutes, filter with a membrane filter, and perform high performance liquid chromatography.
(Column: TSK-gel G-3000SW) was used to quantify albumin.

No. Regarding the herbal extracts 1 to 8, both 99.5% and 50% ethanol extracts showed a high TRS activity inhibition rate. These protein denaturation rates were slightly low in the 99.5% ethanol extract of rosemary (No. 3) and the 50% ethanol extract of Hypericum (No. 4), but were low in general. Therefore,
These herbal extracts showed a high inhibitory effect on TSR activity, and this effect was not due to denaturation of the enzyme protein.

No. Although the lindane extract of 9 has a high TSR activity inhibitory rate, it has a large protein denaturation rate of 92.21% and is considered to have an inhibitory effect due to the denaturation of enzyme protein, which is not preferable for our purposes.

No. Significant TSR for each of 10-15 herbal extracts
No activity inhibitory effect was observed.

The effect of ethanol was not observed in this measurement system.

Furthermore, among the above eight herbal extracts, there are some that exhibit an action of suppressing lipid biosynthesis in the sebaceous glands. That is,
Inhibition of lipid biosynthesis was observed in organ cultures of auricular skin of 10-week-old golden hamsters. The organ culture was carried out by removing the cartilage by peeling the inner skin from a hamster ear piece (6 mmφ), and then [ ¹⁴ C] -sodium acetate 4.4 μm.
Culture medium containing 20 μl of Ci and sample solution (Eagl containing 10% FMS
2 ml of eMEM medium to which penicillin, streptomycin, fungizone, and L-glutamine were added) was floated so that the epidermis side was facing up, and the incubation was carried out at 37 ° C for 6 hours. As a control, the other auricle skin was cultivated in a culture medium containing only the solvent instead of the sample solution. After culturing, wash with physiological saline, put in 2N NaBr solution, leave at 37 ° C for 1 hour, separate dermis and epidermis, put the obtained dermal sheet in 2 ml of chloroform: methanol (2: 1). The lipids were extracted for 12 hours. The inhibition rate of lipid biosynthesis is [ ¹⁴ C]-
The inhibition rate of sodium acetate uptake was determined, and the results are shown in FIG.

Significant suppression of uptake was observed in the hop ( Humulus lupulus L.) extract and the rosemary ( Rosmarinus officinalis L.) extract. These suppression rates are 46.2 each
The values were 1% and 19.89%, respectively, showing a higher inhibition rate than pyridoxine hydrochloride, which is a vitamin generally said to inhibit lipid biosynthesis. This lipid biosynthesis inhibitory action is likely to be mediated by inhibition of TSR activity.

In addition, it is generally known that, in a biosynthesis experiment by incorporation of [ ¹⁴ C] -labeled substance in a culture system, promotion and inhibition of biosynthesis by hormone action do not appear until 12 hours or more after culture (V. Wheatly. and J.Brind ； JI
nvest.Dermatol. 76 293-296, 1981). Therefore, the above-mentioned effects are not mediated by hormones, and there is no concern that serious side effects will occur.

<Effect> As described above, sage ( Salvia officinalis L.), hops ( Humulus lupulus L.), rosemary ( Rosmarinus of
ficinalis L.), Hypericum ( Hypericum erectum Thun)
b.), peppermint (Mentha arvensis L.var. piperascens Mali
nv.), European peppermint ( Mentha piperita L.), chamomile
( Matricaria chamomilla L.), Arnica ( Aunica montan
a .) and thyme ( Thymus vulgaris L.) ethanol extracts exhibit TSR inhibitory action and sebum production inhibitory action. Therefore, select one or two or more of these from the TSR.
As an inhibitor, it can be incorporated into a hair nourishing / hair-growth agent or a hair cosmetic, or a skin external preparation for treating acne or a skin cosmetic. In that case, since the above-mentioned action is not due to protein denaturing action or hormonal action, it is possible to provide a safe cosmetic or the like which is free from skin irritation and side effects.

[Brief description of drawings]

FIG. 1 is a diagram showing inhibition of [ ¹⁴ C] -sodium acetate uptake, which represents the inhibition rate of lipid biosynthesis.

Claims (1)

[Claims] 1. Sage ( Salvia officinalis L.), hop
( Humulus lupulus L.), rosemary ( Rosmarinus offi
cinalis L.), Hypericum ( Hypericum erectum Thun)
b.), peppermint (Mentha arvensis L.var. piperascens Mali
nv.), European peppermint ( Mentha piperita L.), chamomile
( Matricaria chamomilla L.), Arnica montan
a .), Thymus vulgaris L.), and one or more testosterone 5α-reductase inhibitors selected from the ethanol extract.