JPH0524126B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [0001] The present invention relates to a novel 5α-reductase inhibitor. To be more specific, Randium domesticus
Lansium domesticum Jack var. Duku) or Lansium (Lansium domesticum Jack var. Duku) or Lansium domesticum Jack var.
The present invention relates to a 5α-reductase inhibitor containing an extract obtained from the pericarp of A. Langsat as an active ingredient. [0002] Traditionally, the causes of androgenetic alopecia include (1) hormonal imbalance theory, (2) genetic theory, (3)
Many theories have been proposed, including the theory of poor blood circulation and (4) the nutritional theory, but it has long been suggested that the male hormone testosterone plays an important role in the development of hair. Adachi et al.'s theory [Biochem.
Biophys.Res.Commun., 41 , 884 (1970)], testosterone biosynthesized in the testicles is present in hair follicles, hypertrophic glands, etc. in the head.
It is converted to dihydrotestosterone by 5α-reductase, and this dihydrotestosterone significantly reduces the activity of adenyl cyclase, thereby inhibiting intracellular cyclic activity.
This results in a decrease in AMP levels, which in turn induces a decrease in energy production in and around the hair and inhibition of protein synthesis. Therefore, it is thought that due to a series of these phenomena, hair in the growth phase transitions to the resting phase, and while this state is repeated, the hair progresses from terminal hair to vellus hair, and finally to male-pattern baldness. In support of this theory, HVSchweikert et al. found that the hair follicles of male-pattern baldness contain more 5α-reductase metabolites, namely dihydrotestosterone, and more than the hair follicles of women and non-bald people. [J.Clin.Endocr., 38, 811]
(1974)]. [0003] In addition to androgenetic alopecia, dihydrotestosterone, which is produced from testosterone by 5α-reductase, also plays an important physiological role in the occurrence and aggravation of acne (▲acne, acne, etc.). It has been reported. i.e. JBHay
compared the metabolic rate of testosterone between affected skin and normal skin in acne patients.
It has been reported that the metabolism of testosterone by 5α-reductase is enhanced in acne-affected areas [Br.J.Dermatol. , 91 , 123 (1974)]. Also
G. Sansone et al. found that the ability to synthesize dihydrotestosterone from testosterone in the affected skin of acne patients was abnormally enhanced by 2 to 20 times that of normal people.
-It is suggested that dihydrotestosterone produced by reductase is largely involved [J. Invest. Dermatol. , 56, 366 (1971). [0004] Based on these findings, the present inventors strongly inhibited the action of 5α-reductase,
As a result of intensive research to find a 5α-reductase inhibitor that is useful for the treatment and/or prevention of diseases caused by overproduction of dihydrotestosterone, such as alopecia and acne, we found that 5α-reductase inhibitors, a member of the Meliaceae family, The present invention was completed based on the discovery that an extract obtained from the pericarp of a plant can achieve the purpose. Therefore, the present invention relates to a 5α-reductase inhibitor containing as an active ingredient an extract obtained from the pericarp of Zuku root. [0005] Zuku and Langsa are shrubs of the Meliaceae family that are widely distributed throughout Southeast Asia, especially Indonesia, and bear fruit during the rainy season from January to April.
In particular, the fruit of the Zuku tree is widely loved as a fruit, and its pulp is delicious, but the thick skin of the fruit contains a large amount of milky fluid, and has traditionally been said to be poisonous [EJHCorner, Kiyohiko Watanabe, Illustrated illustration] See "Tropical Products Collection" p. 403 (1969) Hirokawa Shoten]. [0006] Although not all of the components of the pericarp have been elucidated, the main ones have been isolated and their structures have been determined. The following four types of compounds have been reported as main extracts. [Chemical 1] [Chemical] [Chemical 2] [Chemical] [0007] Lansic Acid
is the most abundant component in the dried pericarp, and its isolation and structure were determined from 1967 to 1968 [Tetrahedron Letters, 37 , 3571
(1967) and Ibid., 34 , 3731 (1968)]. Lansoside is contained in a more polar fraction than lanjitsuacid, and since it is a glycoside, lansoside A and B, respectively.
and named C [Tetrahedron Letters,
23, 1349 (1982) and J.Org.Chem., 48 , 4462
(1983)]. [0008] The extract included in the present invention includes all components extracted from the dried pericarp of Zuku and Langsa and mixtures thereof, and may also include components whose structures have not yet been isolated or determined. This is also supported by the examples described below. That is, in Experimental Example 1, the ethanol extract from Zuku (presumed to be a mixture of several components with known and unknown structures) has a strong 5α-reductase inhibitory effect, and this fact indicates that the ethanol extract from Zuku (presumed to be a mixture of several components with known and unknown structures) has a strong 5α-reductase inhibitory effect. This suggests the possibility that there are also strong inhibitory effects. A preferred extract is an ethanol extract. [0009] The pharmacological effects of the extract included in the present invention have not been reported at all so far. For some isolated components, landioside, landoside A inhibits the contraction of guinea pig ileal tissue induced by leukotriene _D4 ;
Landioside B and C are Landioside A
It has only been reported that the inhibitory activity is only about 1/10 of that of [J.Org.Chem., 48 ,
4462 (1983)]. The 5α-reductase inhibitory effect of the present invention has a completely different mechanism of action from the inhibitory effect on leukotriene _D4 , and therefore the 5α-reductase inhibitory effect of the present extract
Reductase inhibitory action is a useful pharmacological action discovered for the first time by the present inventors. [0010] Methods for the extraction, isolation and purification of the extracts included in the present invention, in particular landiac acid and landiside A, B and C, can be found in the aforementioned literature, Tetrahedron Letters and J.Org.
Chem., but the extracts of the present invention are not limited to those obtained by the methods described therein. Also, in the literature, extraction is performed using zuku as a raw material, but it can also be extracted from the peel of langsa in the same way. [0011] To briefly explain the extraction, isolation and purification method, first, the dried peel of Zuku or Lanza is crushed and soaked in a suitable solvent for several hours to several weeks, or using a Soxhlet extractor etc. Extract. The extract is isolated into fractions enriched in each component using a suitable separation means, such as cold silica gel, column or high performance liquid chromatography, and further optionally subjected to the chromatography and/or crystallization methods described above. This can be repeated repeatedly to isolate and purify specific components. The solvent used in each operation may be any solvent suitable for the respective operation, such as pentane, hexane,
Aliphatic hydrocarbons such as heptane and cyclohexane, aromatic hydrocarbons such as benzene, toluene and xylene, halogenated hydrocarbons such as methylene chloride, chloroform and carbon tetrachloride, alcohols such as methanol and ethanol, water, Examples include diethyl ether, petroleum ether, ethyl acetate, acetone, and a mixed solvent of two or more thereof. [0012] The extract included in the present invention has a 5α-reductase inhibitor action, so it can be used in mammals,
It is particularly useful for treating and/or preventing diseases caused by excessive production of dihydrotestosterone by 5α-reductase in humans. Such diseases include, for example, alopecia including androgenetic alopecia, and acne. [0013] To use the extract included in the present invention for the above purpose, it is usually administered systemically or locally, orally or parenterally. The dosage varies depending on age, body weight, symptoms, therapeutic effect, administration method, treatment time, etc., but for example, for the treatment and/or prevention of alopecia and acne, it is usually 10 μg to 10 μg per adult. 50mg, preferably 100μg
It is administered transdermally in the range of ~5 mg once to several times a day. Of course, as mentioned above, the dosage varies depending on various conditions, so there are cases where it is sufficient to use an amount smaller than the above-mentioned dosage range, and there are also cases where it is necessary to administer the dosage beyond the range. [0014] Solid compositions for oral administration according to the present invention include tablets, powders, granules, and the like. In such solid compositions, one or more active substances are present in at least one inert diluent, such as lactose, mannitol, dextrose, hydroxypropylcellulose, microcrystalline cellulose, starch, polyvinylpyrrolidone,
Mixed with magnesium aluminate metasilicate. The compositions may contain additives other than inert diluents in conventional manner, such as lubricants such as magnesium stearate and disintegrants such as fibrin calcium gluconate. Tablets or pills may be coated with a film of gastric or enteric substances such as white sugar, gelatin, hydroxypropyl cellulose, hydroxypropyl methyl cellulose phthalate, or may be coated with two or more layers, if necessary. Also included are capsules of absorbable materials such as gelatin. Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solution suspensions, syrups, elixirs, etc., and commonly used inert diluents, such as Contains purified water and ethanol. In addition to an inert diluent, this composition contains a wetting agent,
It may also contain adjuvants such as suspending agents, sweetening agents, flavoring agents, aromatics, and preservatives. [0016] Other compositions for oral administration include one or more active substances,
Sprays formulated by methods known per se are included. [0017] Injections for parenteral administration according to the present invention include sterile aqueous or non-aqueous solutions;
Includes suspending agents and emulsifying agents. Examples of aqueous solutions and suspensions include distilled water for injection and physiological saline. As non-aqueous solutions and suspensions,
Examples include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, alcohols such as ethanol, and polysorbate 80. Such compositions may further contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. These are sterilized, for example, by filtration through bacteria-retaining filters, by incorporation of sterilizing agents, or by irradiation. They can also be prepared as sterile solid compositions and dissolved in sterile water or sterile injectable solvents before use. Other compositions for parenteral administration include topical solutions containing one or more active substances and formulated in a manner known per se, liniments such as ointments, rectal administration. These include suppositories for intracavity administration and petussaries for intracavitary administration. [0019] Compositions for transdermal administration include topical solutions such as lotions, tonics, sprays, solutions, suspensions, emulsions, and liniments such as creams, gels, and creams. In such compositions, one or more active substances are present in at least one inert diluent, such as distilled water, lower alcohols such as ethanol, higher alcohols such as cetanol, polyethylene glycol, propylene glycol, etc. polyhydric alcohols such as, celluloses such as hydroxypropyl cellulose, animal and vegetable fats,
Contains petrolatum, wax, silicone, vegetable oils like olive oil, surfactants, zinc oxide, etc. In addition to the diluent mentioned above, this composition also contains wetting agents, suspending agents,
It may also contain auxiliary agents such as fragrances and preservatives. [0020] The present invention will be described in detail below using reference examples, working examples, and experimental examples, but the present invention is not limited to these examples. [0021] Reference example 1 Extraction with ethanol Grind 50g of dried apricot peel, and extract with ethanol.
It was soaked in 200ml and left at room temperature for one week. The mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain 5.5 g of an oily substance. [0022] Reference Example 2 Extraction of Ranjitsuka Acid (Method A) 950 g of dried argon fruit peel was crushed, and using a Soxhlet extractor, it was continuously extracted with n-hexane for 5 hours, and the extract was concentrated under reduced pressure. 120g of oily substance
I got it. The obtained oily substance was purified by silica gel column chromatography. Using a mixture of ethyl acetate and n-hexane (2:8) as the elution solvent, 65 g of the portion containing ranjitsu acid was eluted, and then a mixture of ethyl acetate and n-hexane (1:1) was used as the elution solvent. Gradually increase the proportion of ethyl acetate from the solution and finally change it to only ethyl acetate to obtain the portion 40 containing landiosides A, B, and C.
g (total) was eluted. The part containing the range acid was dissolved in n-hexane and crystallized, and the resulting crude crystals were recrystallized from a mixture of ethanol and water (3:1) to form the range acid having the following physical properties. 23g was obtained as white crystals. [0023] NMR (CDCl ₃ solution): δ = 5.57 (1H, m), 4.48,
4.86, 4.82, 4.79, 4.69 and 4.65 (1H, bs each),
1.81, 1.77 and 1.73 (3H, bs respectively), 0.80 and
0.73 (3H, S each); IR (KBr tablet method): ν=3300
~2500, 1705, 1630 cm ^-1 ; Mass: m/e = 470 (M ⁺ ), 455, 452, 397. [0024] Reference Example 3 Extraction of Ranjitsuka Acid (Method B) 5.5 kg of dried and pulverized Azalea pericarp was extracted with ethyl acetate.
1211 was cold-soaked overnight (3 times) and ethyl acetate was distilled off from the combined solution 361 to obtain 920 g of an oily substance. The resulting oil was dissolved in n-hexane, extracted with 20% aqueous potassium hydroxide (3 times), the aqueous layer was washed with n-hexane (2 times) to remove neutral substances, and then extracted with water The layer was acidified with concentrated hydrochloric acid and then extracted with ethyl acetate. The extract was concentrated under reduced pressure, and the resulting residue was
- Crystallization using hexane gave 110 g of crude crystals of Landzik acid. The mother liquor was subjected to silica gel column chromatography (elution solvent, ethyl acetate: n-
The mixture was purified with hexane (1:9) to obtain 200 g of an oily substance containing lung acid. The obtained oil was crystallized using n-hexane to obtain 15 g of crude crystals of Landzik acid. combined coarse crystals
125 g of the product was recrystallized from a mixture of ethanol and water (3:1) to obtain 105 g of Landzik acid having the same physical properties as those in Reference Example 2. Reference Example 4 Extraction of Landioside A, B, and C 40 g of the portion containing Landioside A, B, and C obtained in Reference Example 2 was added to J.Org.Chem., 48 , 4462.
(1983) to obtain landiosides A, B and C. Example 1 Production of tablets containing ethanol extract 5 g of ethanol extract of Zuku, 200 mg of fibrillar calcium gluconate (disintegrant), 100 mg of magnesium stearate (lubricant) and 4.7 g of microcrystalline cellulose were prepared by a conventional method. Mixing and tableting yielded 100 tablets containing 50 mg of active ingredient in each tablet. Example 2 Production of lotion containing ethanol extract 0.1 g of ethanol extract of Zuku, 1.1 g of hydroxypropyl cellulose (HPC-M: registered trademark, manufactured by Nippon Soda) and several drops of fragrance were mixed to 80% by a conventional method.
The target lotion was obtained by dissolving it in ethanol and making a total volume of 100 ml. Example 3 Production of cream containing ethanol extract A mixture of 4.0 g each of polyethylene glycol-400 and polyethylene glycol-4000 (both registered trademarks, manufactured by NOF) and 0.5 g of cetanol was dissolved by heating at 80°C. After that, 0.1 g of ethanol extract of Zuku was added and sufficiently dissolved, and the mixture was cooled to room temperature. A few drops of fragrance and purified water were added to the mixture and thoroughly stirred to give a total amount of 10 g to obtain the desired cream. [0029] Reference Example 5 Manufacture of a lotion containing Ranjitsu Acid [0029] 0.1 g of Ranjitsu Acid, 1.1 g of hydroxypropyl cellulose (HPC-M: registered trademark, manufactured by Nippon Soda) and several drops of fragrance were mixed in 80% ethanol using a conventional method. The desired lotion was obtained by dissolving the lotion in a total volume of 100 ml. [0030] Reference Example 6 Manufacture of a cream containing Ranjitsu Acid A mixture of 4.0 g each of polyethylene glycol-400 and polyethylene glycol-4000 (both registered trademarks, manufactured by NOF) and 0.5 g of cetanol was heated to 80°C. After dissolving, 0.1 g of Langzik acid was added, sufficiently dissolved, and cooled to room temperature. A few drops of perfume and purified water were added to the mixture and thoroughly stirred to give a total amount of 10 g to obtain the desired cream. [0031] Experimental Example 1 Inhibitory effect on 5α-reductase [0032] (1) Experimental method J. Shimazaki et al.'s method [Endocrinol, Japan.
18, 179 (1971)]. In other words, 4g of male rat prostate is 3 times the volume of
The mixture was homogenized with 0.1M HEPES (PH7.4) containing 0.25M sucrose and centrifuged (3000 rpm for 10 minutes). The precipitate was suspended in 10 ml of the above buffer and centrifuged again (3000 rpm for 5 minutes). The resulting precipitate was suspended in 3 ml of the above buffer to obtain an enzyme solution. [0033] Enzyme activity was measured using [4- ¹⁴ C]-testosterone (1.5 nmol, 1.5 × 10 ⁵ cpm), NADPH
(0.5 μmol), the above enzyme solution (0.03 ml), and a total volume of 0.1 ml of the reaction solution containing various concentrations of the analyte at 37°C.
and incubated for 60 minutes. The enzyme reaction was stopped by adding 0.4 ml of a mixture of chloroform and methanol (1:2), followed by centrifugation (3 minutes at 2000 rpm), and 50 μ of the resulting supernatant was spotted on a silica gel thin layer plate. Separation was performed using a mixture of methanol and acetic acid (99.2:0.6:0.2). The plate was subjected to autoradiography, and the radioactivity of the generated dihydrotestosterone was measured using a TLC scanner, and the enzyme activity inhibition rate was calculated. The results are shown in Table 1. [0034] (2) Results [Table 1] ■■■ Turtle shell [0012] ■■■ [0035] From the above results, the ethanol extract and Ranjitsuka acid have a very strong 5α-reductase inhibitory effect. On the other hand, Landiside A, B, and C did not have the same effect as Landitsuac acid, but at 500 μg/ml, 20
It showed a strong inhibitory effect of ~60%. Based on this result, extracts from the pericarp of Zuku and Langsa are
It was concluded that it has 5α-reductase inhibitory activity,
Next, we will use Ranjitsuka acid as a representative example of the active ingredient of the extract to examine its effectiveness in the following animal experimental models and clinical trials, which may be expected to be effective in improving or preventing disease conditions by inhibiting 5α-reductase. It was investigated. Experimental Example 2 Hair growth effect on genetically alopecia mice 3-week-old genetically alopecia mice (HRS/J
-hr-) In 5 cases, a solution containing 1% Landzik acid (0.1 g Landzik acid and HPC-
Dissolve 0.15g of M in 80% ethanol and make the total volume 10ml.
0.1 ml of the solution prepared above was applied to the head once a day. From about the 20th day after the start of application, white vellus hair began to grow in all cases, and from about the 30th day, numerous black terminal hairs were formed. However, 0.1 ml of a solution that does not contain lung acid (a solution prepared by dissolving 0.15 g of HPC-M in 80% ethanol and adjusting the total volume to 10 ml)
When applied in the same manner, no such change was observed. The results are summarized in Table 2. [0037] [Table 2] ■■■ Tortoise shell [0013] ■■■ From the above, the extract contained in the present invention can exhibit excellent hair growth and hair growth effects on mice with genetic alopecia. Proven. Example 3 Hair growth effect on humans Using a lotion (manufactured in Reference Example 5) containing 0.1% lung acid, 8 volunteer patients with androgenetic alopecia (28 to 40 years old) The effects on hair growth and hair growth were investigated. Dosage method: once a day
Approximately 1 ml of the above lotion was sprinkled twice on the thinning hair area of the head and lightly massaged. Efficacy was evaluated by having subjects record symptoms such as dandruff, hair loss, itching, etc. every day before and after administering the lotion, and by closely and objectively observing the depilated area once a month. As a result, symptoms such as dandruff, hair loss, and itchiness were significantly reduced in all subjects from about 2 to 3 days after administration of the lotion, and these effects were sustained throughout the administration period. Furthermore, as a result of periodic observation, 2 out of 8 subjects showed a remarkable hair growth effect (i.e., an increase in the number of hairs, a change from soft hair to terminal hair (a phenomenon in which the hair becomes thicker and harder) 4 months after administration. ) and hair growth promotion), and a significant hair thickening effect (i.e., hair growth promotion) was observed in 4 patients, but no effect was observed in the remaining 2 patients. [0039] From the above, the extract contained in the present invention is not only effective against alopecia mice, but also dandruff, which is noticeable in human male pattern alopecia.
It has been proven that it has the effect of preventing hair loss, itching, etc., and of increasing hair growth. [0040] Example 4 Effect on human acne 15 healthy men and women (15 to 25 years old) suffering from acne
We investigated the effects of Ranjitsukashitsu on acne. When an appropriate amount of a cream containing 1% Ranzitsuac acid (manufactured in Reference Example 6) was applied topically to the acne-affected area twice a day, in the morning and evening, in all cases 1 to 3 days after application. Almost complete disappearance of acne was observed within days. [0041] From the above, it has been proven that the extract included in the present invention has a dramatic therapeutic effect on human acne as well. Example 5 Acute toxicity test 5 week old JCL-ICR male mice (body weight: 24~
Ranjitsuka acid dissolved in physiological saline containing 10% ethanol and 0.4% Tween 80 (registered trademark) was administered into the tail vein to 28 g of mice, and the presence or absence of death was observed for 7 days. As a result, 60mg/Landitsuka
In the intravenous administration of Kg, there were no deaths among the 10 cases, and therefore the LD ₅₀ value was 60 mg/Kg or more. [0043] From the above, it was found that the toxicity of the extract included in the present invention is extremely low. Further, in Experimental Examples 3 and 4, there were no side effects (for example, skin irritation such as itching or swelling), which proved that the extract contained in the present invention can be used safely as a pharmaceutical.

Claims (4)

[Claims] 1. A 5α-reductase inhibitor containing as an active ingredient an extract obtained from the pericarp of Zuku or Langsa. 2. The 5α-reductase inhibitor according to claim 1, wherein the active ingredient is an extract extracted from the pericarp of Zuku with ethanol. 3. The 5α-reductase according to claim 1 or 2, wherein the disease caused by 5α-reductase is alopecia.
Reductase inhibitor. 4. The 5α-reductase according to claim 1 or 2, wherein the disease caused by 5α-reductase is acne.
Reductase inhibitor.