JPH0791185B2 - Antiandrogen - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION Industrial Field of the Invention The present invention relates to pharmaceuticals, in particular anti-androgens.

(Prior art) An anti-androgen action is an action that reduces the androgen activity of testosterone, which inhibits the binding between 5α-dihydrotestosterone (5α-DHT) and an androgen receptor, and increases testosterone by 5α-DHT. It is brought about by inhibiting the action of an enzyme that reduces to.

Antiandrogens currently used for the treatment and prevention of androgen-dependent diseases such as hirsutism, convulsions, male pattern baldness, benign prostatic hyperplasia, prostate tumor, precocious boys, etc. The compound of the present invention is an active ingredient.

(Problems to be Solved by the Invention and Objects of the Invention) Conventional anti-androgens have a steroid-based compound as an active ingredient. The reality is that there are restrictions.

Therefore, it is an object of the present invention to provide a new type of antiandrogen which does not have a steroid type nucleus.

(Means and Actions for Solving Problems and Achieving Purpose) According to the present invention, the above problems are 1,3,5,8-tetrahydroxyxanthone, 1,3,6,8-tetrahydroxyxanthone, At least 1 selected from 1,3,5-trihydroxyxanthone, 1,3,6-trihydroxyxanthone, 1,3,8-trihydroxyxanthone, 1,3-dihydroxyxanthone and 1,6-dihydroxyxanthone The above object is achieved by solving an anti-androgen agent containing a compound of a class as an active ingredient.

That is, the present inventors, as a result of repeated studies to find a novel anti-androgen that does not have a steroid type nucleus, 1,3,5,8-tetrahydroxyxanthone,
1,3,6,8-tetrahydroxyxanthone, 1,3,5-trihydroxyxanthone, 1,3,6-trihydroxyxanthone, 1,3,8-trihydroxyxanthone, 1,3-dihydroxyxanthone and 1 , 6-dihydroxyxanthone
The inventors have confirmed that such substances are used and completed the invention.

Each of the above-mentioned compounds, which are the active ingredients of the anti-androgen agent according to the present invention, is known per se and is contained in some plants, and in this case, it can be obtained by extraction and purification by a conventional method. For example, 1,3,5,8-tetrahydroxyxanthone (conventional name: desmethylveridiforin) is contained in senburi, murasakisen, dog senburi, niitakasen, chiretta grass, etc. It can be extracted and purified by the method described in "Pharmaceutical Journal", Vol. 89, pages 410-417, 1969.

The above-mentioned compounds, which are the active ingredients of the anti-androgen agent according to the present invention, can be produced by chemical synthesis because of their relatively simple structures. For example 1,3,6-
Trihydroxyxanthone is synthesized using resorcinol and 2,4,6-trihydroxybenzoic acid according to the method of PK Grover et al., That is, “J. Chem. Soc.” Page 3982-3985, 1955. be able to.

(Dosage Form and Dose) The anti-androgen agent according to the present invention is not limited in dosage form, and includes tablets, powders, fine granules, granules, capsules, oral liquids, suppositories, and skin agents (liniment agents, lotions, Cream for external use or ointment) and the like, and this formulation can be carried out by a conventional method. The dose varies depending on the type of disease, symptoms, dosage form, age of the patient, etc.
It is appropriate to administer 2-10 mg / day and 0.005-25 mg / day for suppositories in 1 to 4 divided doses. In addition, the active ingredient in the skin agent
It is preferable to prepare a preparation containing 0.00001-0.1% by weight, and to administer this by applying it several times a day depending on the symptoms.

(Examples, etc.) Next, the present invention will be described in more detail with reference to drug efficacy test examples and formulation examples.

Drug efficacy test example 1 (androgen receptor binding inhibitory action) This test is a method of Takayasu et al. “J. Steroid Bioche
m. ”Vol. 19, pp. 1141-1146, 1983.

a) Preparation of Androgen Receptor Solution Male Syrian hamsters were castrated and after 16 hours flank gl
The sebaceous glands of and were removed, and 50 mM Tris-HCl buffer (pH 7.
4), 1.5 mM-EDTA, 1 mM-DTT, 10 mM-Na ₂ MoO ₄ and 10%
After homogenizing the sebaceous glands with a 5-10 volume solution containing (W / V) glycerol, 3000 rpm, 0-4
Centrifuge for 10 minutes at ℃ to collect the supernatant.
The supernatant was collected by centrifugation at 0 rpm and 0-4 ° C for 1 hour. The supernatant obtained in the second time is used as an androgen receptor solution.

b) Measurement of androgen receptor binding inhibitory activity 1 nM
[ ³ H] -R1881 (methyltrienolone, 86.0 Ci / mmol)
After incubating a mixed solution (total amount 150 μl) of 1 μM triamcinolone acetonide, the receptor solution according to the above item a) and various specimen samples at 0 ° C. for 16 hours,
500 μl of a solution containing 0.5% activated carbon and 0.05% dextran T-70 was added and left at 0 ° C. for 10 minutes, and then centrifuged at 3000 rpm for 10 minutes to obtain a supernatant. This supernatant 300
After collecting μl and mixing with liquid scintillation cocktail, [ ³ H]-
The specific total amount of R1881 was measured, the obtained data was introduced into the following equation to determine the inhibitory activity as the inhibitory rate, and from this inhibitory rate
The 50% inhibitory concentration was calculated.

Inhibition rate (%) = [(cs) / c] x100 c: Receptor protein and [ ³ H] -R1 without addition of sample
Specific binding to 881 s: Receptor protein and [ ³ H] -R188 when sample is added
Specific binding amount with 1 c) Results The results are shown in Table 1 below.

Sample: A; 1,3,5,8-Tetrahydroxyxanthone B; 1,3,6,8-Tetrahydroxyxanthone C; 1,3,5-Trihydroxyxanthone D; 1,3,6-Trihydroxyxanthone E; 1,3,8-Trihydroxyxanthone F; 1,3-Dihydroxyxanthone G; 1,6-Dihydroxyxanthone Drug efficacy test example 2 (5α-reductase inhibitory action) a) Preparation of 5α-reductase standard Is adjusted by the method of JD Wilson et al.
l. Chem. ", Vol. 246, pp. 2544-2593, 1971, according to the method described below.

The prostate was excised from Wistar male rats (body weight: 260-290 g) and the capsules and adipose tissue of the prostate were removed to obtain about 480 mg of prostate per animal. A 0.88 M sucrose-1.5 mM CaCl ₂ solution was added at a rate of 3.3 ml per 1 g of this prostate, and the mixture was ground using Ultratrax (manufactured by Janke & Kunke) and a Teflon homogenizer. The resulting homogenate is filtered through gauze and the filtrate is 500x
After centrifuging at 10g for 10 minutes, remove the supernatant and use 1g of prostate
The above sucrose-CaCl ₂ solution was added to the sediment at a ratio of 4.1 ml per 1 to suspend the sediment, and the suspension was suspended at 550 xg for 1
After centrifuging for 0 minutes, the supernatant was removed. This operation, that is, the procedure of suspending the precipitate, centrifuging, and washing the precipitate by removing the supernatant was repeated 5 times. To the precipitate thus obtained, a buffer solution A having the following composition was used.
The mixture was added at a ratio of 1 and suspended, and then sonicated.

Buffer A: 0.01 M Tris-HCl buffer (pH 7.4), 50 μM EDTA, 5 mM MgCl ₂ and 500 μM mercaptoethanol.

After sonication, the solution was centrifuged at 12000 xg for 10 minutes and the precipitate was collected. The following buffer solution B was added to the sediment at a ratio of 0.3 ml per 1 g of the used prostate, the resulting suspension was further sonicated, and then centrifuged at 12000 xg for 10 minutes, and then the supernatant (primary supernatant) Was collected. Regarding the precipitate separated by centrifugation, buffer B was added in the same manner as above, ultrasonic treatment and centrifugation were carried out to collect a secondary supernatant, and this supernatant and the previously collected primary supernatant were collected. A sample of 5α-reductase (hereinafter simply referred to as “enzyme solution”) was obtained by merging with Sei.

Buffer B: 0.3 M NaCl and _{0.1M Na 2 HPO 4 -NaH 2 PO} 4 (pH7.0).

The above operations were carried out under the temperature condition of 0-4 ° C.

After performing the above operations, the enzyme solution was obtained from 30 rats.
11 ml was obtained. When the protein concentration of this enzyme solution was measured by the Lowry method, it was 5.1 mg / ml.

b) 5 by high performance liquid chromatography-fluorescence analysis
Measurement of α-reductase activity inhibition In this test, the fluorescent labeling of steroids and the analytical operation by high performance liquid chromatography were carried out by referring to the method of Goto et al. (JP-A-58-57356).

In the case of measuring 5α-reductase activity, conventionally, a radiolabeled testosterone has been generally used as a substrate, but in this test, JD described above was used.
The 4-pregnene-17α, 21-diol-3,20-dione used in the method of Wilson et al. Was used as a substrate.

The operation method is as follows.

0.583 μM 4-pregnene-17α, 21-diol-3,20-dione, various sample samples (sample concentration: 100 μM), 100
μM NADPH and 0.04M sodium phosphate buffer (pH 6.6)
And a mixed solution (total amount: 1 ml) with 200 μl of the enzyme solution described in the above item a) were incubated at 35 ° C. for 1 hour. Then, ethyl acetate was added to stop the enzymatic reaction, and the substrate and its reduction product, 5α-pregnane-17α, 21-diol-3,20-dione, were extracted from the reaction solution. The extract was concentrated to dryness and 1 was added to the residue.
-0.87 μmol of anthroyl nitrile and 3.6 of quinuclidine
μmol and 200 μl of acetonitrile were added and reacted at 60 ° C. for 30 minutes. The solvent was removed under a nitrogen stream, the residue was dissolved in hexane, and then the extract root 1 column (Merc
(manufactured by Company k) to remove excess labeling reagent and impurities, and then eluting with dichloromethane,
Substrate and its reduction product, 5α-pregnane-17
A compound in which the hydroxy group at the 21st position in α, 21-diol-3,20-dione was 1-anthroylated was obtained.

These compounds were subjected to high performance liquid chromatography (HPLC) under the conditions shown below, and separated and quantified to give 4-pregnene-17α, 21-diol-3,20-dione as 5α-pregnane-17α, 21. The amount reduced to the -diol-3,20-dione was measured.

HPLC conditions: Device: Shimadzu LC-4A (manufactured by Shimadzu), Detector: Shimadzu fluorescence spectrophotometer RF-530 (manufactured by Shimadzu), EX379nm, EM459nm, column: COSMOSIL 5C18 (4.6 x 100 mm), mobile phase: water / Methanol (14:86), flow rate; 1.0 ml / min., Temperature; 25 ° C.

The obtained data was introduced into the following equation, and the inhibitory activity was calculated as the inhibition rate.

Inhibition rate (%) = (1-a / b) × 100 a: 5α-pregnane-17α, 2 when a sample is added
Amount of 1-diol-3,20-dione produced a: 5α-pregnane-17 without addition of analyte sample
Amount of α, 21-diol-3,20-dione produced c) Results The results are shown in Table 2 below.

Samples: A: 1,3,5,8-tetrahydroxyxanthone, B: 1,3,6,8-tetrahydroxyxanthone, C: 1,3,5-trihydroxyxanthone, D: 1,3,6- Trihydroxyxanthone, E: 1,3,8-trihydroxyxanthone, F: 1,3-dihydroxyxanthone, and G: 1,6-dihydroxyxanthone Formulation Example 1 (lininiment agent) Formulation: 1,3,5,8 -Tetrahydroxyxanthone 2.5 mg Tragant 50 g Glycerin 30 ml Ethanol 100 ml Purified water Remaining amount 1000 ml Collect ethanol in a mortar and add 1,3,5,8-tetrahydroxyxanthone to dissolve it. Add and mix, then add glycerin and mix, further mixing while adding 500 ml of purified water to form a paste,
After that, purified water was further added to adjust the total amount to 1000 ml and mixed to prepare a liniment agent. Formulation example 2 (lotion) Formulation: 1,3,5,8-tetrahydroxyxanthone 0.5 mg Hydroxypropyl cellulose 1 g Macrogol 400 10 ml Ethanol 10 ml Purified water The rest 100 ml 1,3,5,8-tetra Dissolve hydroxyxanthone in ethanol, add Macrogol 400 and mix well,
Hydroxypropyl cellulose was added to and mixed with this, and purified water was added to adjust the total amount to 100 ml, followed by treatment with a vacuum homogenizer to produce a lotion.

Reference example (composition for formulation) 1,3,5,8-tetrahydroxyxanthone was dissolved in 100 times amount of ethanol, and 5 times amount of dextrin was added to this and mixed well to form a uniform product, and dried. Then, a pharmaceutical composition was obtained.

Although dextrin is used as a carrier in the above-mentioned pharmaceutical composition, it may be replaced by other known pharmaceutical carriers such as starch, lactose, light anhydrous silicic acid, and magnesium aluminometasilicate. it can. Formulation Example 3 (cream for external use or ointment) Prescription: Composition for formulation (Reference Example) 30 (g) Diethyl sebacate 8 Whale wax 5 Polyoxyethylene oil ether sodium phosphate 6
Sodium benzoate 0.5 Vaseline balance 100 The external preparation cream or ointment was manufactured by the above-mentioned prescription and a conventional method. Formulation Example 4 (Suppository) Formulation: Composition for formulation (Reference Example) 60 (mg) Oil-based base (cacao butter) 1640 1700 per piece A suppository was prepared by the above-mentioned formulation and by a conventional method. . Formulation Example 5 (Powder) Formulation: Pharmaceutical composition (Reference Example) 30 (mg) Lactose 800 Corn starch 170 1000 per packet A powder was produced by the above-mentioned formulation and by a conventional method. Formulation Example 6 (Granule) Formulation: Composition for formulation (Reference Example) 30 (mg) Lactose 754 Corn starch 200 Hydroxypropylcellulose 16 1000 per 1 package Granules were produced according to the above-described formulation and by a conventional method. Formulation Example 7 (Tablets, coated tablets and sugar-coated tablets) Formulation: Composition for formulation (reference example) 30 (mg) Crystalline cellulose 40 Lactose 52.5 Corn starch 30 Magnesium stearate 7.5 160 per tablet According to the above formulation and according to a conventional method Manufactured.

On the other hand, a part of the tablets thus obtained was made into a water-soluble film-coated tablet with the following coating formulation (weight ratio) and by a conventional method.

Water-soluble film coating formulation: Hydroxypropyl cellulose 40 Macrogol 6000 10 Titanium oxide 3 Talc 5 Purified water 942 Furthermore, for some of the above tablets, the following sub-coating formulation and coloring formulation (weight ratio) and sugar-coated tablets were prepared by a conventional method. I said.

Sub-coating formulation: Sucrose 40 Gelatin 0.5 Gum arabic 1.4 Precipitated calcium carbonate 22 Talc 15.6 Purified water 20 Coloring formulation: Sucrose 10 Titanium oxide 73 Lake dye 56 Purified water 5 Formulation example 8 (hard capsule) Formulation: Pharmaceutical composition ( Reference Example) 30 (mg) Lactose 104 Corn starch 40 Hydroxypropyl cellulose 16 190 per 1 capsule Lactose and corn starch are added to and mixed with a pharmaceutical composition, and an aqueous solution of hydroxypropyl cellulose is added and kneaded. Granules were produced by an ordinary method using an extrusion granulator. The granules were filled into a gelatin hard capsule to manufacture a hard capsule.

(Effect of the Invention) 1,3,5, which is the active ingredient of the anti-androsen agent according to the present invention,
8-tetrahydroxyxanthone, 1,3,6,8-tetrahydroxyxanthone, 1,3,5-trihydroxyxanthone, 1,3,6-trihydroxyxanthone, 1,3,8-trihydroxyxanthone, 1, Unlike the drugs conventionally used for the treatment and prevention of androgen-dependent diseases, 3-dihydroxyxanthone and 1,6-dihydroxyxanthone do not have a steroid type nucleus, and therefore have high safety in use.

Since these compounds themselves are known compounds, can be extracted and purified from various plants, have a simple structure and can be chemically synthesized, the anti-androgen agent according to the present invention is provided at a low price. be able to.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Eiichi Kato 3-36 Sennari-dori, Nakamura-ku, Nagoya, Aichi (72) Inventor Seiko Oishi 1-9, Matsukaze-cho, Gifu-shi, Gifu Inventor Kunio Yagi 2-21 Nishizato-cho, Meito-ku, Nagoya-shi, Aichi

Claims (1)

[Claims] 1. A 1,3,5,8-tetrahydroxyxanthone,
1,3,6,8-tetrahydroxyxanthone, 1,3,5-trihydroxyxanthone, 1,3,6-trihydroxyxanthone, 1,3,8-trihydroxyxanthone, 1,3-dihydroxyxanthone and 1 An anti-androgen agent, which comprises at least one compound selected from 6,6-dihydroxyxanthone as an active ingredient.