JPS62246519A - 5alpha-reductase inhibitor - Google Patents

Abstract

PURPOSE:To provide a 5alpha-reductase inhibitor containing an extract of liverwort or liverwort callus as an active component and effective for suppressing the excessive production of dihydrotestosterone causing apopecia, acne, prostatomegaly, etc. CONSTITUTION:Liverwort or liverwort callus is extracted with an organic solvent, preferably a 5/1-1/5 (vol) mixture of a hydrophilic organic solvent and a hydrophobic solvent and the extract is used as a 5alpha-reductase inhibitor. The 5alpha-reductase is useful as a preventive and remedy for diseases caused by excessive production of dihydrotestosterone, e.g. male apopecia, acne, prostatomegaly, etc., by inhibiting the conversion of testosterone to dihydrotestosterone. It is administered systemically or locally by oral or parenteral administration.

Description

Detailed Description of the Invention [Field of Industrial Application] The present invention relates to a novel SO-reductase inhibitor, and more specifically, Liverwort te is a 5a-reductase inhibitor containing an extract from Liverwort callus as an active ingredient. O regarding agents [Conventional technology] Traditionally, the causes of male deafness are (1) Hormone imbalance theory? ) genetic theory, a> blood circulation failure theory,
(4) Although many theories have been proposed, including the nutritional theory, it has long been suggested that the male hormone testosterone plays an important role in hair growth. That is, testosterone, which is biosynthesized in the kidney, is converted to dihydrotestosterone in the head VC by 5α-reductase present in hair follicles, sebaceous glands, etc. This dihydrotestosterone significantly decreases the activity of adenyl cyclase, leading to a decrease in the intracellular cyclic oxidation level, resulting in a decrease in energy production in and around the hair and inhibition of Q-resynthesis. It is thought that this induces androgenetic alopecia progresses.

Dihydrotestosterone, which is produced from testosterone by 5α-reductase, is thought to be involved in the occurrence and exacerbation of acne, and enlargement of the prostate (
J, Steroid Biochemistry*l 1
t609 (1979)].

[Problem that the invention seeks to solve]

However, no effective drug for these diseases, particularly an effective drug based on inhibition of dihydrotestosterone production, has been found, and its development has been desired.

[Means for solving problems]

In view of the above circumstances, the present inventors conducted intensive research to find a 5α-reductase inhibitor that suppresses the excessive production of dihydrotestosterone, which causes alopecia, acne, and prostatic hyperplasia. The inventors discovered that extracts and the like meet the purpose, and completed the present invention.

That is, the present invention provides a 5α-reductase inhibitor containing an extract from liverwort callus as an active ingredient.

As the liverwort from which to extract the active ingredient of the 5α-reductase inhibitor of the present invention, any liverwort belonging to the order Liverwort can be used. For callus of liverworts, a medium in which liverworts can grow, such as Muras
hig@/5took mediums Murashige/
A synchronized culture cell collection cultured using Skoog medium or the like is used.

To extract the active ingredients from liverworts or liverwort callus, these dried products are soaked in a suitable organic solvent for several hours.
This is done by soaking for several days or by extracting using an extractor such as a Soxhlet extractor. Although the extract thus obtained can be used as it is as an active ingredient of a 5α-reductase inhibitor, it is also possible to use an appropriate separation means such as silica gel or high-performance liquid chromatography to isolate the extract with high activity. It can also be fractionated and used.

The extraction solvent may be either a hydrophilic organic solvent or a hydrophobic organic solvent;
1 to 115 (volume ratio) mixed solvent is preferred. Examples of hydrophilic organic solvents include methanol, ethanol, ZoronoQ-nol, and acetone. Examples of hydrophobic organic solvents include pentane, hexane, cyclohexane, benzene, toluene, xylene, methylene chloride, chloroform, carbon tetrachloride, diethyl ether, petroleum ether, and ethyl acetate. Any of these can be used as a mixture of two or more types. The results of an investigation of the inhibitory effect of the extract thus obtained on 5α-reductase, the pharmacological effect based on this effect, and the toxicity are shown below.

α) Inhibitory effect on 5α-reductase Male rats (body weight 300-4509) were subjected to cervical vertebrae, and the prostate gland was aliquoted and homogenized in 3 volumes of 20 rrM phosphate buffer (pH 7.5) containing 0.32 M sucrose. The precipitate obtained by centrifugation (140ρ00t×60min) was solubilized with 10mM phosphate buffer containing 40% glycerin containing 511/lId zogitonin, and then centrifuged (1
The supernatant was used as an enzyme solution. (T, Liaug et al. J, 1llio1
．． chem,, 256 e 7998 * 198
1) Measurement of enzyme activity was performed using 50 μj of the above enzyme solution, (1,
2゜6 e 7 a 16 e l 7- ``H]-tes) containing 0.25 i/10 t4 Nl of PH (50 hermitage) and various concentrations of analyte or non-radioactive testosterone. A total volume of 50 ILl of the reaction solution was incubated at 37°C for 1 hour.To this, ethyl acetate was added to stop the reaction.The ethyl acetate fraction was collected, and after distilling off the solvent with nitrogen, it was diluted with 50 g of methanol. After re-dissolution, 20 m of the solution was taken and subjected to high-performance liquid chromatography, and the amounts of testosterone and dihydrotestosterone were measured using a W detector and an isotope detector, thereby determining the amount of enzyme activity inhibition.

Table 1 Extracts 1 to 3 were produced according to Reference Examples 1 to 3.
Oe 45-54 a1983)
Day C3) 1 The hair on the back of the mouse was clipped with clippers, and the lotion l- shown in Example 2 was applied to this area once a day. Table 2 shows the results of a comparison of the number of days required until hair growth was observed in all areas compared to a control using only ethanol.

Table 2 0) Effect on Ni-mei-pi 15 healthy men and women were divided into left and right based on the midline of the forehead, and one of them received the cream shown in Example 3, and the other received only the base. It was applied twice a day, and the number of pimples and the amount of sebum secreted at the site were measured every day. After one week, a decrease in the number of pimples and a decrease in the amount of sebum secreted was observed in the cream VC. It was also found to be effective against vulgaris.

(4) When performing acute toxicity tests by oral administration using toxic mice, no deaths were observed even when 10r/IlJ was administered.

Since the extract included in the present invention has a 5α-reductase inhibitory effect, it is useful for treating and/or preventing diseases caused by excessive production of dihydrotestosterone by 5α-reductase in salivary mammals, especially humans. Such diseases include, for example, alopecia including androgenetic alopecia, acne, and benign prostatic hyperplasia.

To use the extract included in the present invention for the above-mentioned purposes, it is usually administered systemically or locally, daily or parenterally.

The dosage varies depending on age, body weight, symptoms, therapeutic effect, administration method, treatment time, etc., but for the treatment and/or prevention of benign prostatic hyperplasia, it is usually 1 kg to IP once per adult.
, preferably administered orally in the range of 20 mg to 200 mg, once to several times a day, or once per adult.
C100/Jg ~IQ□ay.

It is preferably administered parenterally (preferably intravenously) once to several times a day within the range of 9 to 10 doses per day. For the treatment and/or prevention of alopecia and acne, it is usually 10 μg to 5019, preferably 1100 μg per adult.
It is administered transdermally once to several times a day in the range of IJ to 5 strokes.

Of course, as mentioned above, the dosage varies depending on various conditions, so there may be cases where an amount smaller than the above dosage range is sufficient,
There may also be cases where it is necessary to administer over the range.

Solid formulations for oral administration according to the present invention include tablets, pills, powders, granules, and capsules. In such a solid preparation, in addition to the extract as the active ingredient, K such as whey, mannitol, grape thorn, hydroxypropyl cellulose, microcrystalline cellulose, starch,
? It may contain an inert diluent such as ribinylpyrrolidone and magnesium aluminate metasilicate; a surface lubricant such as magnesium stearate; and a disintegrant such as cellulose calcium gluconate. For tablets, pills may be coated with a film of gastrointestinal or enteric substances such as sandalwood, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose, phthalate, etc., or they may be coated with two or more layers. Good too.

Liquid preparations for oral administration include emulsions, solutions,
Examples include suspensions, syrups, and elixirs. Such liquid preparations may contain, in addition to the active ingredients and inert diluents, adjuvants such as wetting agents and suspending agents, sweeteners, flavoring agents, aromatics, preservatives, etc. Other formulations for oral administration include sprays and the like.

Typical preparations for parenteral administration according to the present invention include injections. In order to form an injection containing the active ingredient of the present invention, an aqueous medium such as distilled water for injection or physiological saline; propylene glycol; -lyethylene glycol, vegetable oils such as olive oil, alcohols such as ethanol, ?
It can be obtained by dissolving, suspending or emulsifying it in a non-aqueous medium such as Linorpate 80. Injectables may further contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Sterilization measures required for injections include (1) filtration through an S cutilia retention filter, addition of a bactericidal agent, and irradiation. These can be prepared as sterile solid compositions and dissolved in sterile water or sterile injectable solvents before use.

Other preparations for parenteral administration include topical s solutions;
Ointments such as ointments, lotions, tonics, sprays, suspensions, and emulsions; main agents for rectal administration: pessaries for intravaginal administration, etc. Especially for alopecia,
Preferable preparations for treating and preventing acne include lotions, tonics, sprays, solutions, and ointments. In addition to the active ingredients, these preparations contain distilled water; lower alcohols such as ethanol; higher alcohols such as cetanol;
polyhydric alcohols such as ethylene glycol and propylene glycol; celluloses such as hydroxyzorobyl cellulose; animal and vegetable fats; petrolatum; wax; silicone; vegetable sources such as olive oil; surfactants; oxidation Diluents such as zinc, as well as wetting agents, suspending agents,
Auxiliary agents such as fragrances and preservatives can be added.

[Action and effect of the invention]

The 5α-reductase inhibitor of the present invention inhibits the conversion of testosterone to dihydrotestosterone, thereby preventing and preventing diseases caused by excessive production of dihydrotestosterone, such as androgenetic alopecia, acne, and benign prostatic hyperplasia. Useful as a therapeutic agent.

〔Example〕 Next, the present invention will be explained with reference to reference examples and examples.

Reference Example 1 Extraction from Liverwort Plants 50 tons of dried liverwort plants were crushed and immersed in chloroform/methanol l/2v/ma 200dK for 3 days.

Filter the mixture and concentrate the filtrate to an oil (Extract 1)
I got 1o and 4f.

Reference Example 2 Improved medium of Muraahige/Skoog extracted from Liverwort callus (MSK-
2) Freeze-dried product of nine liverwort callus 5
0t, chloroform/methanol l/2v/v 20
0ml! It was immersed in VC for 3 days. The mixture was filtered and the filtrate was concentrated to obtain 20.4 t of oil (extract 2).

Reference Example 3 Separation of Flavonoid Fraction from Liverwort Callus Extract Using a preparative thin-layer chromatography plate, first remove the carotenoid and chlorophyll fractions with petroleum ether as a developing solvent, then use chloroform as a developing solvent, The flavonoid fraction (extract 3) was carefully selected.

This flavonoid fraction was about 10% of the spot amount.

Example 1 Tablet tearing Reference example t, Z, extract lt shown in 3, cellulose calcium gluconate (disintegrant) 200 cm, magnesium stearate (wetting agent) x00+y and crystalline cellulose 8.7
1 was mixed in a conventional manner and compressed into tablets to obtain 10 m of the extract.
100 tablets containing f were obtained.

Example 2 Manufacture of 0-silon Extracts shown in Reference Examples 1 and 2.3 0.19s Hydroxypropyl cellulose i, l? A lotion was obtained by dissolving a few drops of perfume in 80% ethanol in a conventional manner to make a total volume of 100 ml.

Example 3 cream manufacturing Extract 0.11P shown in Reference Example 1.2.3.

A mixture of polyethylene glycol 4000 s, or and cetanol 0.52 was dissolved by heating at 80°C, and then cooled to room temperature. Add water and stir thoroughly to make a cream.

that's all

Claims (1)

[Scope of Claims] 1. A 5α-reductase inhibitor containing an extract from liverworts or liverwort callus as an active ingredient. 2. The 5α-reductase inhibitor according to claim 1, wherein the extract from liverworts or liverwort callus is an extract using an organic solvent. 3. 5α according to claim 1, wherein the extract from liverworts or liverwort callus is a mixed solvent of a hydrophilic organic solvent and a hydrophobic organic solvent at a volume ratio of 5/1 to 1/5.
- Reductase inhibitors.