JPH0353287B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to novel 5α-reductase inhibitors. More specifically, the present invention relates to a 5α-reductase inhibitor containing landiac acid or landoside A, B, or C as an active ingredient. Many theories have been proposed for the cause of male pattern hair loss, including (1) hormonal imbalance theory, (2) genetic theory, (3) blood circulation deficiency theory, and (4) nutritional theory. It has long been suggested that the male hormone testosterone plays an important role in hair development. Adachi et al.'s theory experimentally proved the causal relationship between testosterone and androgenetic alopecia at the biochemical level [Biochem.Biophys.Res.
Commun., 41 , 884 (1970)], testosterone biosynthesized in the testes is converted to dihydrotestosterone (5α-reductase) in the head by 5α-reductase present in hair follicles, fatty glands, etc. dihydrotestosterone),
This dihydrotestosterone significantly reduces the activity of adenyl cyclase, leading to a decrease in intracellular cyclic AMP levels, resulting in a decrease in energy production in and around the hair and suppression of protein synthesis. do. Therefore, due to these series of phenomena,
It is thought that the 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 bald people have more
It has been reported that metabolites of 5α-reductase, such as dihydrotestosterone, are present in large amounts [J. Clin. Endocr., 38 , 811 (1974)
See also]. In addition to male pattern baldness, testosterone can also cause
It has been reported that dihydrotestosterone produced by 5α-reductase plays an important physiological role in the occurrence and aggravation of acne (acne, acne, etc.). That is, JB Hay et al. compared the metabolic rate of testosterone between the affected skin of acne patients and normal skin, and reported that the metabolism of nestosterone by 5α-reductase was accelerated 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 has been suggested that dihydrotestosterone produced by reductase is largely involved [J. Invest. Dermatol., 56 , 366 (1971)]. Based on these findings, the present inventors determined that 5α-
We have conducted intensive research to find a 5α-reductase inhibitor that strongly inhibits the action of reductase and is useful for the treatment and/or prevention of diseases caused by overproduction of dihydrotestosterone, such as alopecia and acne. As a result, the present invention was completed by discovering that lungiac acid or lungiside A, B, or C can achieve the object. Lansic Acid (Lansic Acid) is Lansium domesticum Jack var.
It is the most abundant component in the extract obtained from the pericarp of Duku, and its isolation and structure were determined from 1967 to 1968 [Tetrahedron Letters, 37 , 3571 (1967) and the same magazine, 34 , 3731 (1968)]. Lansioside is contained in a more polar fraction than lansioside, and since it is a glycoside,
They were named landiosides A, B and C, respectively [Tetrahedron Letters, 23 , 1349 (1982)
and J.Org.Chem., 48 , 4462 (1982)]. Its structural formula is as follows. Landioside A (R=N-acetyl-β-D
- Glucosamine) Landiside B (R = β-D-glucose) Landiside C (R = β-D-xylose) There have been no reports on the pharmacological effects of landiside acid. For lungioside, lungioside A is leukotriene D ₄
However, it has been reported that landiosides B and C have only about one-tenth the inhibitory activity of landioside A. There is [J.Org.
Chem., 48 , 4462 (1983)]. of the present invention
The 5α-reductase inhibitory effect is caused by leukotrienes.
The inhibitory effect on _D4 has a completely different mechanism of action, and therefore the 5α-reductase inhibitory effect of the extract of the present invention is a useful pharmacological effect discovered for the first time by the present inventors. . Methods for extraction, isolation and purification of landiac acid and landoside A, B and C included in the present invention are described 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 these documents, extraction is performed using zuku as a raw material, but lansium (Lansium domesticum
It can also be extracted in the same way from the pericarp of domesticum Jack var.Langsat). 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. Zuku fruit is especially popular as a fruit, and its pulp is delicious, but its thick skin contains a large amount of milky fluid.
Legend has it that it is poisonous [EJH
Corner, Kiyohiko Watanabe, Illustrated Tropical Plant Collection, p. 403 (1969) Hirokawa Shoten]. To briefly explain the extraction, isolation, and purification method, first, the dried fruit skin of Zuku or Lanza is crushed and soaked in a suitable solvent for several hours to several weeks, or extracted using a Soxhlet extractor, etc. . isolating the extract into fractions enriched in each component using suitable separation means, such as silica gel, columns or high performance liquid chromatography;
Furthermore, the above-mentioned chromatography and/or crystallization method can be repeated as desired to isolate and purify the target component. Since the compounds included in the present invention have a 5α-reductase inhibitory effect, they are useful for treating and/or preventing diseases caused by excessive production of dihydrotestosterone by 5α-reductase in mammals, particularly humans. Such diseases include, for example, alopecia, including androgenetic alopecia, and acne. To use the compounds included in the present invention for the above purposes, they are usually administered systemically or locally, orally or parenterally. Dosage depends on age, weight, symptoms,
Although it varies depending on the therapeutic effect, administration method, treatment time, etc., for example, it can be used to treat alopecia and acne, and/or
Alternatively, in the case of prophylaxis, the dose per adult is usually 10 μg to 50 mg, preferably 100 μg to 5 mg, administered transdermally 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. Solid compositions for oral administration according to the present invention include tablets, powders, granules, and the like. In such fixed compositions, one or more active substances are present in at least one inert diluent, such as lactose, mannitol, glucose, hydroxypropylcellulose, microcrystalline cellulose, starch, polyvinylpyrrolidone, aluminum metasilicate. mixed with magnesium acid. 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, solutions, suspensions, syrups, elixirs, etc., and commonly used inert diluents, such as purified water. , including ethanol. In addition to inert diluents, the compositions may also contain adjuvants such as wetting agents and suspending agents, sweetening agents, flavoring agents, aromatic agents, and preservatives. Other compositions for oral administration include sprays containing one or more active substances and formulated in a manner known per se. Injections for parenteral administration according to the present invention include sterile aqueous or non-aqueous solutions, suspensions,
Includes emulsifying agents. Examples of aqueous solutions and suspensions include distilled water for injection and physiological saline. Examples of non-aqueous solutions and suspensions include propylene glycol, polyethylene glycol,
These include vegetable oils like olive oil, alcohols like 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:
It contains one or more active substances and is formulated according to methods known per se. These include liquid preparations for external use, liniments such as soft creams, suppositories for intrarectal administration, and petals for intravaginal administration. Compositions for transdermal administration include external solutions such as lotions, tonics, sprays, solutions, suspensions, and emulsions, and topical solutions 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 hydroxypropylcellulose, animal and vegetable fats, petrolatum, wax, silicone, vegetable oils such as olive oil, surfactants, zinc oxide, etc. In addition to the diluents mentioned above, the composition may also contain adjuvants such as wetting agents, suspending agents, fragrances, and preservatives. Hereinafter, the present invention will be explained in detail with reference to Reference Examples, Examples, and Experimental Examples, but the present invention is not limited to these Examples. Reference Example 1 Extraction of Ranjitsuka acid (Method A) 950g of dried argon fruit peel was crushed, extracted continuously with n-hexane for 5 hours using a Soxhlet extractor, and the extract was concentrated under reduced pressure to obtain an oily substance. 120g
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 crystallized from a mixture of ethanol and water (3:1) to obtain the range acid having the following physical properties. 23g was obtained as white crystals. NMR (CDCl ₃ solution): δ = 5.57 (1H, m), 4.88,
4.86, 4.82, 4.79, 4.69 and 4.65 (1H each,
bs), 1.81, 1.77 and 1.73 (3H, bs), 0.80 respectively
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. Reference Example 2 Extraction of Ranjitsuka Acid (Method B) 5.5 kg of dried and crushed argon fruit peel was mixed with 12 ethyl acetate.
After cold soaking overnight (3 times), ethyl acetate was distilled off from the combined solution 36 to obtain 920 g of an oily substance. The obtained oily substance was dissolved in n-hexane, extracted with a 20% aqueous potassium hydroxide solution (3 times), and the aqueous layer was extracted with n-hexane.
Neutral materials were removed by washing with -hexane (twice), and the aqueous layer was then acidified with concentrated hydrochloric acid and extracted with ethyl acetate. The extract was concentrated under reduced pressure, and the resulting residue was crystallized using n-hexane to obtain 110 g of crude crystals of Landzik acid. The mother liquor was purified by silica gel column chromatography (eluent: ethyl acetate:n-hexane = 1:9) to obtain 200 g of an oily substance containing Landzik acid. The obtained oil was crystallized using n-hexane to obtain 15 g of crude crystals of Landzik acid. 125 g of the combined crude crystals were recrystallized from a mixed solution of ethanol and water (3:1) to obtain 105 g of Landzik acid having the same physical properties as in Reference Example 1. Reference Example 3 Extraction of Landioside A, B, and C 40 g of the portion containing Landioside A, B, and C obtained in Reference Example 1 was extracted from J.Org.Chem., 48 , 4462.
(1983) to obtain landiosides A, B and C. Example 1 Manufacture of tablets containing Ranjitsu Acid 5 g of Ranjitsu Acid, 200 mg of fibrillar calcium gluconate (disintegrant), 100 mg of magnesium stearate (lubricant) and microcrystalline cellulose
Mix 47g and press into tablets using a conventional method, each tablet contains 50mg.
100 tablets containing the active ingredient were obtained. Example 2 Manufacture of lotion containing Ranjitsu Acid 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 dissolved in 80% ethanol by a conventional method. The desired lotion was obtained in a total volume of 100 ml. Example 3 Manufacture of a cream containing Ranjitsu Acid After heating and dissolving a mixture of 4.0 g each of polyethylene glycol-400 and polyethylene glycol-4000 (both registered trademarks, manufactured by NOF Corporation) and 0.5 g of cetanol at 80°C. , 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. Experimental example 1 Inhibitory effect on 5α-reductase (1) Experimental method J. Shimazaki et al.'s method [Endocrinol,
Japon., 18 , 179 (1971)]. i.e. male rat prostate 4
0.1M containing 3 times the volume of 0.25M sucrose
The mixture was homogenized with HEPES (PH7.4) and then 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) and add 3 ml of the above buffer to the resulting sediment.
was added and suspended to obtain an enzyme solution. 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 were incubated at 37° C. 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. Methanol and acetic acid (99.2:0.6:
0.2) was used for separation. The plate was subjected to autoradiography, 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 the table. (2) Results

[Table] From the above results, Landitsuac acid has a very strong 5α-reductase inhibitory effect, while Landiside A, B, and C did not show the same effect as Landitsuka acid. However, at 500 μg/ml, it showed a strong inhibitory effect of 20-60%. Using the landing gear,
We investigated the effectiveness in the following animal experimental models and clinical treatments in which inhibition of 5α-reductase is expected to be effective in improving or preventing disease conditions. Experimental Example 2 Hair growth effect on genetically alopecia mice 3-week-old genetically alopecia mice (HRS/J
-hr-) A solution containing 1% radic acid (0.1 g of radic acid and HPC-M
0.15g of the solution was dissolved in 80% ethanol to make a total volume of 10ml, and 0.1ml of the solution 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, when 0.1 ml of a solution containing no Landzik acid (a solution prepared by dissolving 0.15 g of HPC-M in 80% ethanol and adjusting the total volume to 10 ml) was applied in the same manner, no such change was observed. I couldn't help it. The results are summarized in the table below.

[Table] From the above, it was proven that the compounds included in the present invention exert excellent hair growth and hair growth effects on genetically alopecia mice. Experimental Example 3 Hair Growth Effect on Humans Using a lotion (manufactured in Example 2) containing 0.1% Ranjitsu Acid, hair growth was observed on 8 volunteer patients with androgenetic alopecia (28 to 40 years old). and its effectiveness in hair growth. 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. The effectiveness was evaluated by having the subjects record symptoms such as dandruff, hair loss, and itching 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. Also, as a result of regular observation, 4 days after administration.
At the 2nd month, notable hair growth effects (i.e., an increase in the number of hairs, a change from soft hair to terminal hair (a phenomenon in which hair becomes thicker and harder), and promotion of hair growth) were observed in 2 out of 8 test subjects. , a significant hair thickening effect (i.e. promotion of hair growth) was observed in 4 people,
No effect was observed in the remaining two patients. Based on the above, the compounds contained in the present invention are not only effective against alopecia mice, but also show remarkable effects on preventing dandruff, hair loss, itching, etc. and hair growth and hair growth effects in human male pattern alopecia. This has been proven. Experimental 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 Example 3) was applied topically to the acne-affected area twice a day, in the morning and evening, all cases showed 1 to 3 days after application. Almost complete disappearance of acne was observed within days. From the above, it has been proven that the compounds included in the present invention have a dramatic therapeutic effect on human acne as well. Experimental example 5 Acute toxicity test 5 week old JCL-ICR male mice (body weight: 24~
28g) was injected intravenously into the tail vein with Landjitsuka acid dissolved in physiological saline containing 10% ethanol and 0.4% Tween 80 (registered trademark), and the presence or absence of death was determined after 7 days.
Observed for days. As a result, Landscape Assist 60
In intravenous administration of mg/Kg, there were no deaths among the 10 cases, and therefore the LD ₅₀ value was 60 mg/Kg or higher. From the above, it was found that the compounds included in the present invention have very low toxicity. In addition, in Experimental Examples 3 and 4, there were no side effects (e.g. irritation to the skin such as itching and swelling).
It has been demonstrated that the compounds included in the present invention are sufficiently safe to be used as pharmaceuticals.

Claims (1)

[Scope of Claims] 1. A 5α-reductase inhibitor containing Landitsu acid or Landiside A, B or C or a mixture thereof as an active ingredient. 2. The 5α-reductase inhibitor according to claim 1, wherein the disease caused by 5α-reductase is alopecia. 3. The 5α-reductase inhibitor according to claim 1, wherein the disease caused by 35α-reductase is acne.