JPH0368518A - Testosterone 5alpha-reductase inhibitor - Google Patents

Abstract

PURPOSE:To obtain the title inhibitor useful for treating prostatic carcinoma, male type alopecia, acne, etc., comprising a specific flavonoid contained in Morus Ihou of crude drug. CONSTITUTION:The title inhibitor containing a compound shown by formula I [R1 is 3-methyl-2-butenyl or group shown by formula II), formula III (R2 is 3-methyl-2-butenyl or 3-hydroxy-3-methylbutyl), formula IV (R3 is H or 3,7- dimethyl-2,6-octadienyl; R4 is H or group shown by formula V), formula VI, formula VII (R5 is group shown by formula VIII or formula IX) obtained from an extract of Morus Ihou or a homologous plant thereof in an organic solvent as an active ingredient.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention relates to a drug that has a testosterone 5α-reductase inhibitory effect and is useful for the treatment of i4 glandular hypertrophy, androgenetic alopecia, acne, etc. .

[Prior Art and Problems] In general, substances having male hormone effects are collectively referred to as androgens, and testosterone is considered to be the most important androgen in humans.

Testosterone is a true testicular hormone, and is said to exhibit hormonal effects after being taken into cells of target organs and converted into dihydrotestosterone by 5α-reductase.

Currently, the typical diseases caused by overproduction of dihydrotestosterone include androgenetic alopecia, prostatic hypertrophy,
There are acne etc. In general, therapeutic agents for these diseases include:
Drugs that modify the steroid structure have been administered, but there are problems such as the appearance of undesirable hormonal adverse effects and the mode of administration. Furthermore, at present, there is no therapeutic agent for the above-mentioned diseases that does not have a steroid structure that has been put to practical use yet.

Therefore, it has been desired to develop a testosterone 5α-reductase inhibitor that has fewer side effects, is easy to administer, and has excellent efficacy.

[Means for Solving the Problem] As a result of extensive research in order to provide an excellent testosterone 5α-reductase inhibitor, the present inventors have developed the herbal medicine Mulberry White Bark, which is also used clinically, and its original plant. A certain mulberry (
We discovered that some flavonoids contained in Morus 1hou (such as Morus 1hou) and other reported plants have the above-mentioned inhibitory effect, and completed the present invention ['J1].

That is, the present invention is as shown below.

(1) Testosterone containing a compound represented by the following formula I as an active ingredient 5
α-reductase inhibitor.

(2) The following 2■ (However, R1 is a 3-methyl-2-butenyl group or (1 group ■■ (However, R2 is a 3-methyl-2-butenyl group or a 3-hydroquino-3-methylbutyl group.) Testosterone containing a compound as an active ingredient 5
α-reductase inhibitor.

(3) The following formula■ (4) The following formula■■ (However, R8 is a hydrogen atom or 3,7-dimethyl26
A testosterone 5α-reductase inhibitor containing as an active ingredient a compound represented by the following formula: -octagenyl group, and R4 represents either a hydrogen atom or a group (■).

Testosterone whose active ingredient is the compound represented by (the following margin) 5
α-reductase inhibitor.

(5) Testosterone 5α containing a compound represented by the following formula ■ (wherein R6 represents either group ■ or IX) as an active ingredient
・Reductase inhibitor.

Hereinafter, the compounds of Formula 1, ■, ■, ■, and ■ will be collectively referred to as compounds of the formula.

For example, the following methods can be used to obtain the compound of the formula.

After degreasing the roots or whole plants of mulberry or its unlicensed plants with 11-hexane as necessary, benzene, ethyl acetate,
Extract with an organic solvent such as ethanol or methanol, remove the solvent from the extract, and dissolve the resulting residue in an appropriate solvent such as methanol, benzene, or ethyl acetate.
Porous polymers such as Amberlite XAD-2, Diaion HP-20, MCI Gel CHP 20 P, Sephadex, etc. are used as an elution solvent with at least one selected from ethanol, acetic acid, chloroform, ethyl acetate, n-hexane, acetone, and benzene. Sephadex such as Ll-1-20, reversed-phase silica gel, silica gel, polyamide, activated carbon, cellulose, etc. are used as a carrier at least once for column chromatography, and the target components are confirmed and separated using thin layer chromatography. It can be obtained by drawing.

Alternatively, t11 may be produced by recrystallization using a suitable solvent such as benzene, ethyl ether, n-hexane, acetone, methanol, or ethanol, as the case may be.

A specific example of the production of the compound of the formula is shown below. 4-0 Specific Example 1 Dry the root bark of cultivated mulberry (Morus Ihou (dry weight: 26 kg)), defatt with n-hexane, and extract sequentially with benzene and ethyl acetate. did. The solvent of the extract was distilled off under reduced pressure to obtain 300 g of Hensen's extract and 1.3 to 9 of the ethyl acetate extract.

Next, benzene extract 3009 was dissolved in methanol to obtain 200 g of methanol-soluble extract. This methanol-soluble extract +001i1 was subjected to column chromatography using silica gel and eluted with a benzene-methanol system. The benzene eluted area was searched by thin-layer chromatography, and the part containing spots that emitted dark blue fluorescence under ultraviolet irradiation was collected. This part was purified by preparative thin-layer chromatography and re-purified from benzene-n-hexane. By crystallization, 200 colorless plate crystals were obtained.

The physical and chemical properties of this colorless plate-like crystal are described in the literature [Heterocycles, 9.1593 (1978
)] was consistent with that of Marberofuran A described.

Concrete example 2 Concrete example! In column chromatography using silica gel, the 1% methanol-benzene eluate was purified by preparative thin layer chromatography and recrystallized from benzene-ethyl ether to obtain pale yellow prism nucleus crystals 14+y.

The physical and chemical properties of this pale yellow prism-like product are described in the literature [Ch.
em, Pharm, Bull, , 26.1453 (1
978)] was consistent with that of Kwanon C described.

Specific Example 3 In the column chromatography using silica gel in Specific Example 1, the 1% methanol-benzene eluate was purified by preparative thin layer chromatography, and ethyl ether-n-
Recrystallized from hexane, colorless prismatic crystals 230■
I got it.

The physical and chemical properties of this colorless prism nucleus crystal are described in the literature [Pla
The results were consistent with those of Kwanone E described in [Nta Med, 42, 79 (1981)].

Concrete Example 4 In the column chromatography using silica gel in Concrete Example 1, the 2% methanol-benzene eluate was searched for by preparative thin layer chromatography, and the part that showed green color in the ferric chloride reaction was collected and reconstituted with methanol. Crystallization was carried out, and 60 counterfeit crystals (9) were obtained.

The physical and chemical properties of this yellow needle crystal are described in the literature [Chem, P.
harm, Bull, 26.1453 (197g)]
It corresponded to that of the described oxydihydromorsin.

Specific Example 5 In the column chromatography using silica gel in Specific Example 1, the 0.1% methanol-benzene eluate was searched for by preparative thin layer chromatography, and the part showing green color due to the ferric chloride reaction was collected, and the methanol The soluble portion was further subjected to column chromatography using silica gel, the water-saturated benzene eluted portion was searched for by preparative thin layer chromatography, the portion showing green color in the ferric chloride reaction was collected, and ethyl ether-〇 By recrystallizing from -hexane, 2.4 g of pale yellow prism nucleus crystals were obtained.

The physical and chemical properties of this pale yellow prism nucleus crystal are described in the literature [Ch.
em, Pharm, Bull, 26, 1394 (19
78)] was consistent with that of morsin described.

Specific example 6 3009 was taken from the ethyl acetate extract of specific example 1,
It was subjected to silica gel column chromatography using benzene-methanol as an eluent. 5% methanol-
The benzene eluate was further treated with benzene-acetone (3:2
), the Rr value was 4.4, the color was green due to ferric chloride reaction, and 20-30
After a few seconds, the brown discolored area was collected.

This was again subjected to column chromatography using silica gel, and 25% aceton/n-hegisan elution part 12
I got a 9.

From this, 59 was taken and purified by preparative thin layer chromatography to obtain 2 g of pale yellow powder.

The physical and chemical properties of this pale yellow powder are described in the literature [letter
ocyles, 14, 1943 (1980)]
It corresponded to that of the described quasin I-1.

Specific Example 7 The same operation as in Specific Example 6 was performed to obtain a pale yellow powder of 30Q.

The physical and chemical properties of this pale yellow powder are described in the literature [PIanta
Med, 47, 151 (1983)].

Specific Example 8 In column chromatography using benzene-methanol as the eluent in Specific Example 6, the 3% meta/-Luhensen eluate was searched with a thin layer of silica gel [1. Part 20 that emits
I got a 9.

From this, 29 was taken, separated by preparative thin layer chromatography, and further purified by Sephadex L l-120 column chromatography, resulting in colorless powder 3.
Obtained 0m9.

The physicochemical properties of this colorless powder are described in the literature [Chem.

Pharm, Bull, J3, 3195 (1985)
] It corresponded to that of Marberofuran G described.

Specific Example 9 3 h of commercially available mulberry bark was extracted sequentially with n-hexane, benzene, and methanol, and the solvent of the methanol extract was distilled under reduced pressure to obtain 300 g of methanol extract.

This methanol extracted extract was dissolved in ethyl acetate and the solvent was distilled off to obtain ethyl acetate soluble extract 709. This ethyl acetate soluble portion was subjected to column chromatography using silica gel, and the 20-41!5% acetone-benzene eluate portion 8? I got it. This was further purified by preparative thin layer chromatography to obtain 2 g of yellow powder.

The physical and chemical properties of this yellow powder are described in the literature [11e Lerocycles, 16, 2141. (
1981)] was consistent with that of Sangenone C described in [1981].

Next, the fact that the compound of the formula has a testosterone 5α-reductase inhibitory effect will be explained with reference to experimental examples.

Experimental Example 1 <Preparation of Testosterone 5α-Reductase> An 11-week-old F344 male rat was dissected and the prostate gland was removed. The obtained prostatic capsule and adipose tissue were removed, and a 0.25M N-Eucrose solution was added thereto at a ratio of 51 d1g, followed by homogenization using a cell crusher (POLYTRON). The obtained homogenate was filtered through gauze,
The filter limbs were further homogenized using a Dounce homogenizer, and after sonication, a microsomal fraction was obtained by centrifugation. This microsomal fraction was 3.37
The enzyme was suspended in 0.05M potassium-phosphate buffer (PI-46, 6) containing 0.25M sucrose, and this was used as an enzyme solution.

<Measurement of testosterone 5α-reductase inhibitory activity> The above enzyme solution 50Ii! 0.05M potassium-phosphate buffer (pH 6.6), NADPH (final concentration 1x10-'M), glucose-6-phosphate (5x
l O-3M) glucose-6-phosphate dehydrogenase (0.51U), bovine serum albumin (0°1%)
, [4-"C]-testosterone, and the compound of the formula were added to bring the total amount to 600. After reacting this mixture at 30°C for 60 minutes, IN hydrochloric acid was added to terminate the reaction. Next, testosterone and 5α were added as carriers. - Dihydrotestosterone was added, and the reaction product was extracted with ethyl acetate 1,jd. After concentrating the extracted substance, testosterone and 5α-dihydrotestosterone were separated by thin layer chromatography, and 5α-dihydrotestosterone was extracted using a nontyrene counter. Testosterone radioactivity was measured.

In addition, except that the compound of the formula was not added, the inhibition rate was calculated using the following formula in the same manner as in the case of Nisoku, using shino as a rhyme.

-B Inhibition rate (%) - X I OO A: 5α when the compound of formula is not added Dihydrotestosterone production amount B: 5α when adding the compound of formula Nohydrotestosterone production amount The results are shown in Table 1.

From the results shown in Table 1 and above, the inhibitory effect of the compound of the formula on testosterone 5α-reductase was confirmed.

Next, an acute toxicity test of the compound of the formula was conducted using ICR male mice, and it was found that oral administration of 1.097 ksl L>
There were no cases of death, and the drug was highly safe.

Thus, the compound of the formula has extremely low toxicity and high safety.

Next, the dosage and formulation of compounds of formula will be described.

The compound of the formula can be administered to animals and humans as is or together with conventional pharmaceutical carriers.

The dosage form is not particularly limited and may be selected and used as required, including tablets, capsules, granules, fine granules,
Examples include oral preparations such as powders, parenteral preparations such as injections, and cross preparations.

In order to exert the desired effect as an oral agent, although it depends on the age, weight, and level of illness of the patient, the usual adult dosage is 50 mg to 5 mg of the compound of the formula in divided doses several times a day. It seems appropriate to take .

Oral agents include, for example, starch, lactose, sucrose, mannitol,
It is manufactured using conventional methods using carboxymethyl cellulose, corn starch, inorganic salts, etc.

In addition to the excipients mentioned above, binders, disintegrants, surfactants, lubricants, glidants, flavoring agents, coloring agents, fragrances, and the like can be used in this type of preparation as appropriate. Specific examples of each are shown below.

[Binder] Starch, dextrin, gum arabic powder, gelatin,
Hydroxypropyl starch, methylcellulose, sodium carboxylmethylcellulose, hydroxypropylcellulose, crystalline cellulose, ethylcellulose, polyvinylpyrrolidone, macrogol.

[Disintegrant] Starch, hydroquinepropyl starch, carpoquine methylcellulose sodium, carpoquine methylcellulose carnoum, carpoxymethylcellulose, low substituted hydroxypropylcellulose.

[Surfactant] Sodium lauryl sulfate, soybean lecithin, polysaccharide fatty acid ester, polysorbate 80° [Lubricant] Talc, waxes, hydrogenated vegetable oil, polysaccharide fatty acid ester, magnesium stearate, turquoise stearate, stearin Aluminum acid, polyethylene glycol.

[Ryusha accelerator] Light anhydrous silicic acid, dry aluminum hydroxide gel, synthetic aluminum silicate, magnenoum silicate.

The compounds of the invention can also be administered as suspensions, emulsions, solots, elixirs,
These various dosage forms may contain flavoring agents and coloring agents.

In order to exert the desired effect as a parenteral agent, it is usually necessary to administer the compound of the formula intravenously to an adult at a dose of 0.1 to + + g per day, although this will vary depending on the age, weight, and severity of the disease of the patient. , intravenous drip, subcutaneous injection, and intramuscular injection are considered appropriate.

This parenteral preparation is manufactured according to a conventional method, and diluents generally include distilled water for injection, physiological saline, glucose aqueous, distilled water for injection, sesame oil, peanut oil, soybean oil, corn oil, propylene glycol, polyethylene glycol, etc. can be used. Furthermore, bactericides, preservatives, and stabilizers may be added as necessary. In addition, from the viewpoint of stability, this parenteral preparation may be filled into a vial or the like, then frozen, water removed by ordinary freeze-drying techniques, and a liquid preparation prepared from the freeze-dried product immediately before use. Furthermore, isotonizing agents, stabilizers, preservatives, soothing agents, etc. may be added as appropriate.

Other parenteral preparations include liquid preparations for external use, liniments such as ointments, and preparations for intrarectal administration, and are manufactured according to conventional methods.

Next, the present invention will be explained in more detail with reference to Examples.
The present invention is not limited to this.

Example 1 ■Corn starch 449 ■Crystalline cellulose 409 ■Carboquine methylcellulose calcium 5g ■Light silicic anhydride 0.5g ■Magnenium stearate 0.5g ■Compound obtained in specific example 1
+09 Total of 100 pieces ■ to ■ were mixed uniformly according to the above recipe, and compression molded using a tablet machine to obtain one tablet of 2001!9.

This tablet-tablet contains 20 mg of the compound obtained in Gutai Fei 1, and adults should take 10 to 25 tablets a day in several doses.

Example 2 ■Crystalline cellulose 84.5Li■Magnesium stearate 0.59■Carboxomethyl cellulose calcium 59 ■Compound obtained in Example 2 tog total 1009 According to the above recipe, ■, ■, and part of ■ were uniformly mixed. After compression molding, the mixture was pulverized, the remaining amounts of (1) and (2) were added and mixed, and the mixture was compressed and molded using a tablet machine to obtain a portion of 200 Q tablets.

These tablets contain 20 ml of the compound obtained in Example 2, and adults should take 10 to 25 tablets in several doses per day.

Example 3 ■ Crystalline cellulose 49.59 ■ 10% hydroquinpropyl cellulose ethanol solution 35 g ■ Calcium carboxymethyl cellulose 59 ■ Magnesium stearate 059 ■ Compound obtained in Example 3 10 g Total 100 g According to the above recipe ■, ■ and After uniformly mixing (1), making a netting using a conventional method, granulating it using an extrusion granulator, drying and crushing, (2) and (2) were mixed, and compression molded using a tablet machine. mg of ji2 agent was obtained.

These tablets contain Compound 20-9 obtained in Specific Example 3, and are taken by adults in 10 to 25 tablets a day in several doses.

Example 4 ■Corn starch 34.5y ■Magnesium stearate 509 ■Calcium carboxymethylcellulose 59 ■Light anhydrous silicic acid 0.590 Compound obtained in Example 4 10g Total 100g Mix ■~■ uniformly according to the above recipe and compress. After compression molding with a molding machine, it was crushed with a crusher and sieved to obtain granules.

1 g of this granule contains the compound l00R9 obtained in specific example 4.
It contains 2 to 5 g per day for adults, divided into several doses.

Example 5 ■ Crystalline cellulose 55 g ■ IO% hydroxypropyl cellulose ethanol solution 359 ■ Compound obtained in Example 5 Log total 1003 According to the above recipe, ■ to ■ were uniformly mixed and suspended. After going into an extrusion granulator and granulating, the mixture was dried and sieved to obtain granules.

This granule 11? is the compound obtained in specific example 5 + oox
Adults should take 2 to 59 doses in several doses per day.

Example 6 ■Corn starch 89.5 Lj ■Light anhydrous silicic acid 9.5 g ■Compound obtained in Example 6 to, 1 roog Uniformly combine ■ to ■ according to the above recipe, 20011
9 was filled into a No. 2 capsule.

One capsule of this preparation contains 20 mg of the compound obtained in Example 6, and adults should take 10 to 25 capsules in several doses per day.

Example 7 ■ Soybean #Ib 59 ■ Distilled water for injection 89.5 g ■ Soybean phospholipid 2.59 ■ Glycerin 2 g ■ Compound obtained in Example 7 19 Total amount 100 g Dissolve ■ in ■ and ■ according to the recipe in L And this ■
The n1 relatives of (1) and (2) were added and emulsified to obtain an injection.

Claims (5)

[Claims] (1) A compound represented by the following formula I ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (However, R_1 is a 3-methyl-2-butenyl group or a group II ▲There are mathematical formulas, chemical formulas, tables, etc.▼) Testosterone 5α with active ingredient
- Reductase inhibitors. (2) Formula III below ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (However, R_2 represents a 3-methyl-2-butenyl group or a 3-hydroxy-3-methylbutyl group) as an active ingredient. A testosterone 5α-reductase inhibitor. (3) Formula IV below ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (However, R_3 is a hydrogen atom or 3,7-dimethyl-
2,6-octadienyl group, R_4 represents either a hydrogen atom or group V) ▲ Numerical formula, chemical formula, table, etc. are available ▼ Testosterone 5α whose active ingredient is a compound represented by
- Reductase inhibitors. (4) Testosterone 5α whose active ingredient is a compound represented by the following formula VI ▲Mathematical formula, chemical formula, table, etc.▼
- Reductase inhibitors. (5) Formula VII below ▲Mathematical formulas, chemical formulas, tables, etc.▼ (However, R_5 represents either group VIII or IX)
▲There are mathematical formulas, chemical formulas, tables, etc.▼ Testosterone 5α whose active ingredient is the compound represented by
- Reductase inhibitors.