JPH01207256A - Novel cis-(6)-shogaol and remedy for hyperkeratosis containing said compound as active ingredient - Google Patents

Abstract

NEW MATERIAL:cis-(6)-Shogaol[=1-(4-hydroxy-3-methoxyphenyl)-(Z)-4-decene-3- one] expressed by the formula. USE:A medicine and active ingredient of remedy for hyperkeratosis having low toxicity and high safeness. The doze is 20-80mg/adult.day based on amount of the active ingredient and the active ingredient is administered several times. PREPARATION:A dried rhizome of Zingiberis Rhizoma is extracted with an organic solvent such as ether or methanol or mixed solvent thereof to give an extract, which is then applied to a column chromatography using a silica gel as carrier and developed with an organic solvent such as n-hexane or mixed solvent thereof to afford a fraction containing trans-(6)-shogaol, which is further applied to preparative thin-layer chromatography to purify the fraction. The resultant purified product is irradiated with ultraviolet ray using a low-pressure mercury lamp for about 1-3hr to provide the compound expressed by the formula.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention provides a novel system (6) effective for the treatment of keratosis, etc.
- Concerning shogaol.

[Prior Art and Problems] Chinese herbal medicines such as Saiko Kaji Kanpanto, Shoseiryuto and Kakkonba contain zingerone as the main ingredient, (6)- Shogaol, gingerol, etc. are known, and among these, (6)-shogaol has been confirmed to have pharmacological effects such as antipyretic action, analgesic action, and antispasmodic action.

This (6)-shogaol has stereoisomers because it has a double bond in its side chain, but currently only the trans isomer isolated as a natural product is known.

Keratosis represented by psoriasis has been on the rise in recent years, and in psoriasis in particular, the main morphological changes in the skin at the lesion site are thickening and keratosis of the epidermal cell layer due to markedly accelerated turnover of epidermal cells. abnormalities in cell formation, as well as inflammatory reactions in the papillary epidermal layer and fugue migration of polynuclear leukocytes to the epidermal cell layer.

Until now, steroids have been used as therapeutic agents for this keratosis, but serious side effects have become a problem. Furthermore, the use of vitamin Ai conductors (retinoids) is restricted due to the problem of flu malformation, and hitherto there has been no innovative therapeutic agent for this keratosis.

[Means for Solving the Problems] The present inventors have repeatedly conducted research on dry skin and its main ingredients, and as a result of intensive study to develop a drug effective for the treatment of keratosis, the present inventors have developed a novel system (6). )-shogaol and further discovered that this compound has an excellent effect on the treatment of keratosis, and based on this finding, the present invention was completed.

That is, the present invention provides a novel Nsu(6)-shogaol[1-
(4-hydroxy-3-methoxophenyl)-(Z14
-decene-3-onco, and a therapeutic agent for keratosis containing this compound as an active ingredient.

This cis-(6)-shogaol can be obtained by irradiating trans-(6)-shogaol contained in the main or dry matter with ultraviolet rays.

The raw material trans-(6)-shogaol can be obtained, for example, as follows.

Dried rhizomes of ginger are extracted with organic solvents such as ether, ethanol, methanol, etc. alone or in combination to obtain an extract, which is then subjected to column chromatography using silica gel as a carrier. , developed with an organic solvent such as ethyl acetate or a mixture thereof to obtain a fraction containing trans-(6)-shogaol, which is further purified by preparative thin layer chromatography. be able to.

A specific example of the production of trans-(6)-shogaol is shown below.

Specific example] Ginger (Zingiber oCficinale)
2 kg of dried rhizomes of P. roscoe were ground and extracted under reflux with ether 3Q for 7 hours. The extraction residue is further diluted with ether 3
Reflux extraction was carried out with Q twice for 7 hours. Combine the above extracts, remove the ether under reduced pressure, and extract 77.059
I got it. The ether extract 77.059 was added to silica gel 7001? (Merck, Kieselgel
60) was subjected to column chromatography using n-
It was developed with a mixed solvent of hexane and ether while increasing the proportion of ether.

The fraction eluted with 0.6 g of fi-hexane:ether (70:30) and n-hexane:ether (60;
40) The fractions eluted at 1.6 ρ were combined and the solvent was removed to obtain 13.6 g of residue. The residue was again subjected to column chromatography using silica gel, and n-
The fraction eluted with hexane:ether (95:5) was collected, and the residue obtained by removing the solvent was subjected to preparative thin layer chromatography (preparative thin layer chromatography), Kieselg
el 60 P Fzs+: developing solvent, n-hexane;
acetone (7:3)] to give a pale yellow oily substance 0.803
9 (yield 0.04%) was obtained. The physicochemical properties of the pale yellow oily substance were consistent with those of trans-(6)-yogaol described in the literature.

The trans-(6)-shogaol obtained as described above is dissolved in an organic solvent such as methanol or ethanol, and cis-(6)- Obtain shogaol.

Next, the fact that cis-(6)-shogaol has a 5-lipoxygenase inhibitory effect, a cyclooxygenase inhibitory effect, and a keratosis therapeutic effect will be explained using experimental examples.

[Experimental Example 115-Lipoxygenase inhibitory effect RBL-1
IJIM the cultured cells to 5xlO' cells/1d.
50% phosphorus I containing EDTA and 10% ethylene glycol! 2 buffer solution (pH 7,4), and after sonication, 105,000
The supernatant obtained by centrifugation at xG for 60 minutes was used as a 5-lipoxygenase enzyme preparation.

For the reaction, the entire volume was mixed with Q, 6-distillate, 50mM phosphate buffer, 1
511M Calcium Chloride, 20th Edition Indomethacin, 18
mM glutathione, 6xMATPS 10/7M arachidonic acid, enzyme preparation obtained as above and cis-(
6) - Shogaol ethanol solution with a final concentration of 5!
, 0.577111 in a test tube and heat at 37°C.
, and reacted for 10 minutes. After the reaction is completed, acetone 1. ]d
, 100% of 2N formic acid was added to stop the reaction, and then the internal a
Add 0.25M n-butyl-3,5-dinitrobenzoate IOm as Q, add n-hexane t, 5rnl
Extracted with. After concentrating the obtained extract, it was dissolved in methanol, and the amount of 5-HETE therein was analyzed by high performance liquid chromatography [column, TSK-gel ODS-807M
(manufactured by Tosoh Corporation); mobile phase, acetonitrile:water diacetic acid (60:40:0.02); flow rate, 1 d1 minute; detection, ultraviolet light (235 nm)], and this was taken as the enzyme activity. Regarding the results, the inhibition rate at each concentration was calculated using the following formula, and the 50% inhibition concentration (hereinafter referred to as ICa) was determined.

Inhibition rate--x I 00 (%) C: 5 when not containing cis-(6)-shogaol
-H[ETE peak area (corrected by internal standardization) S; 5 when cis-(6)-shogaol is added
-HETE peak area (corrected by internal standard) As a result, the IC5o of cis-(6)-shogaol was 0.83 #M.

[Experimental Example 2] Cyclooxygenase Inhibitory Effect 60 μm of 0.1 M phosphate buffer (pH 7.5) containing 1 geltadeone was added to 7.0 μg of rabbit kidney medulla and homogenized using a Polytron. 9,0OO of this homogenate
After centrifugation at XG for 20 minutes, the supernatant was further centrifuged at 105,
The microsomal fraction was obtained by centrifugation at 000xG for 60 minutes. This was dissolved in O, 1M phosphate buffer (pl+7.5) containing 1 x M glutathione to prepare a cyclooxygenase enzyme preparation.

As the final concentration of the enzyme preparation 15d obtained as above,
4 Geltadeon, IOxM) Liptophan, 0.25
Add stimulated hemoglobin and phosphate buffer (p117.5) and add [1-1"C co-arachidonic acid (5x l
O'dpm) and Nsu (6)-shogaol 4
0.20, l015- and the total amount is 20OA
And so. After incubation at 37°C for 15 min, IN
Hydrochloric acid was added to stop the reaction, prostaglandin E (PGE,) was added as a carrier, and the reaction product was extracted with ether 2d. After concentrating the obtained extract, it was subjected to thin layer chromatography (developing solvent: chloroform; methanol:acetic acid-18:I:I) to separate the products. The area corresponding to PGE that was colored with iodine vapor was scraped off, and PGE and radioactivity were measured using a liquid scintillation counter. From the results, the inhibition rate at each concentration was calculated using the following formula to obtain IC5o.

C: P when cis-(6)-shogaol is not included
GE, production amount S: P when cis-(6)-shogaol is added
As a result, the IC7° of cis-(6)-shogaol was 11.7 vices.

[Experimental Example 3] Growth inhibitory effect on epidermal growth factor (EGF) 24
Fibrolial cells (BALB/C 3T3 ce) were added to the hole plate.
A culture medium containing 10% calf serum (hereinafter referred to as CS D
My name is MEM. ) was sown per hole. After the fibrogenic cells were saturated in a monolayer state, 1% CS DM
Changed to EM and cultured for 12 hours at 37°C under 5% CO1.
0.0 cis-(6)-shogaol per l hole, respectively.
54 and an additional 16 with addition of 50 pg of mouse EGP.
Cultured for hours. Add 0.5μC1150m/well to this
After adding ``H-thymidine'' and culturing for 8 hours, the culture medium was removed, the cells were washed with chilled isotonic phosphate buffer, and 5% cold tric aa acetic acid was added and left for 30 minutes.The supernatant was removed. After that, dissolve in 0.5M sodium hydroxide aqueous solution, 0.5M
The mixture was neutralized with hydrochloric acid, added with a syndylene cocktail, and counted using a liquid scintillation counter. The effect of cis-(6)-shogaol on cell proliferation was investigated by measuring the radioactivity of 'H-thymidine incorporated into the cells with this proliferation. As a result, 639
% growth inhibition was observed.

From this result, it was confirmed that cis-(6)-shogaol has an excellent therapeutic effect on keratosis.

Next, an acute toxicity test of Sosu(6)-Nyogaol was conducted.
When conducted using CR male mice, 500 MW
No deaths were observed after oral administration.

Thus, cis-(6)-shogaol has low toxicity and high safety.

Next, the dosage and formulation of cis-(6)-shogaol, which is the active ingredient of the drug of the present invention, will be explained.

Cis-(6)-shogaol can be administered to animals and humans either neat or with conventional pharmaceutical carriers. The dosage form is not particularly limited and may be selected and used as necessary, and may include oral dosage forms such as tablets, capsules, granules, fine granules, and powders, and parenteral dosage forms such as injections and suppositories. It will be done.

In order to exert the desired effect as an oral agent, the weight of cis-(6)-shogaol is usually 20 to 80 II for adults, although it varies depending on the age, weight, and severity of the disease of the patient.
! It seems appropriate to take 9 in divided doses several times a day.

Furthermore, oral preparations such as tablets, capsules, and granules are manufactured according to conventional methods using, for example, starch, lactose, sucrose, mannitol, carboquine methylcellulose, cornstarch, inorganic salts, and the like.

In addition to the excipients mentioned above, binders, disintegrants, surfactants, lubricants, fluidity promoters, 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.

[Binding agent] Starch, dextrin, gum arabic powder, gelatin,
Hydroquinbrovir starch, methylcellulose, sodium carpoquinomethylcellulose, hydroquinopropylcellulose, crystalline cellulose, ethylcellulose, polyvinylpyrrolidone, macrogol.

[Disintegrant] Starch, hydroquine bropyl starch, carpoquine methylcellulose sodium, carboxymethylcellulose calcium, carboxymethylcellulose, low substituted hydroxypropylcellulose.

[Surfactant] Sodium lauryl sulfate, soybean lentil, sucrose fatty acid ester, polysorbate 80° [Lubricant kotalc, waxes, hydrogenated vegetable oil, sucrose fatty acid ester, magnesium stearate, carunium stearate, aluminum stearate, Polyethylene glycol.

[Fluidity promoter] Light anhydrous silicic acid, dry aluminum hydroxide gel, synthetic aluminum silicate, magnesium silicate.

The drug of the present invention can also be administered as a suspension, emulsion, syrup, or elixir, and these various forms may contain flavoring agents, stabilizers, and coloring agents. Good too.

In order to exert the desired effect as a parenteral agent, it is usually necessary to use 0.25 cis-(6)-shogaol per day for adults, although it varies depending on the age, weight, and severity of the disease of the patient.
Intravenous infusion, intravenous drip infusion, subcutaneous injection, and intramuscular injection up to ~1OQ are considered appropriate.

This parenteral preparation is manufactured according to a conventional method, and generally uses distilled water for injection, physiological saline, aqueous glucose solution, vegetable oil for injection, sesame oil, peanut oil, soybean oil, corn oil, propylene glycol, polyethylene glycol, etc. as a diluent. be able to. Furthermore, a bactericide, a preservative, and a stabilizer may be added as necessary. In addition, from the viewpoint of stability, this parenteral preparation can also be frozen after being filled into a vial or the like, 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 external solutions, ointments such as ointments, omentum for intrarectal administration, etc., 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 in any way by this.

Example 1 The trans-(6)-shogaol 29 shoulder 9 obtained in the specific example was treated with 1. d was dissolved in methanol, and this solution was placed in a quartz cell and irradiated with ultraviolet rays for 1 hour using a low-pressure mercury lamp.

After the reaction, it was subjected to silica gel column chromatography (ether, hexane = 2 = 8) with a 30-fold dip, fractionated into 5d fractions, and the 18th to 22nd fractions were combined to obtain a pale yellow oil having the following physical and chemical properties. Substance cis-(6)-
Shogaol 7, 5119 was obtained.

Infrared absorption spectrum νn+ a x α-1: 3540
．． 1680 Proton nuclear magnetic resonance spectrum (δppm in CDCl5): 0.89 (3H,t, J=7.0Hz).

1.31 (4H, m), 1.42 (2H, m).

2.61 (2[-I, dt, J = 7.0.6.9
Hz).

2.75 (2+1.t, J=7.5l-(Z).

2.86 (211,t, J=7.5LIz).

3.87 (311, s).

6.07 (111, dt, J = 7.0.11.5
Hz).

6.12 (I I-1, dt, J = l, 3, I
I, 5 Hz).

6.68 (I H, dd, J = 1.9.8.0
Hz).

6.70 (l H, d, J = 1.9 Hz).

6.82 (I H, d, J = 8, Of-1z
) 13c-Nuclear magnetism J (Sound spectrum (δppm in CDCl5): 14.0. 22.5. 2B, 9. 29.5° 29
．． 7. 3+, 5. 46.1. 55.9°111.
1. 114.3. 120.9°126.6. 13
3.2. 143.9°146.4. 149.0 Mass spectrum; m/z (%) 276 (M'', 27).

205(27).

+37 (+00), 55 (37) Example 2 ■Corn starch 23.59■Cellulose 159 ■Carboquine methylcellulose calcium 59 ■Light silicic anhydride 0.59■Magnesium stearate 19 ■Nos-6-shogaol 5 Total 509 According to the above-mentioned recipe, ① to ② were uniformly mixed and compressed using a tablet machine to obtain tablets of 200 my in size.

This tablet contains cis-(6)-shogaol 20
19, and adults should take 1 to 4 tablets a day in several doses.

Example 3 ■ Crystalline cellulose 39.59 ■ Magnesium stearate 0.59 ■ Carpoquine methyl cellulose calcium 59 Nsu (6-7 yogaol 5 total 50 g According to the above recipe, mix ■, ■, and part of ■ uniformly, 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 tablets each weighing 200 zg.

This tablet contains cis-(6)-shogaol 20
Contains shoulder 9, and adults should take 1 to 4 tablets a day in several doses.

Example 4 ■Crystalline cellulose 179 ■10% hydroquinepropyl cellulose ethanol solution 259 ■Carboxymethylcellulose calcium 2.79 ■Magnenium stearate 0.39 ■Nsu (6)
-Shogaol 5 total 509 Mix ■, ■, and ■ uniformly according to the above recipe, nettle it using a conventional method, granulate it using an extrusion granulator, dry and crush it, and then mix ■ and ■ Then, compression molding was performed using a key press to obtain tablets with a weight of 200 rq.

This tablet contains cis-(6)-shogaol 20
mg, and adults should take 1 to 4 tablets several times a day.

Example 5 ■Corn starch 43g ■Magnesium stearate 0.5g ■Carboxymethyl cellulose calcium 59 ■Light silicic anhydride 0.59 cis-6-shogaol 19 Total 50g Mix ■~■ uniformly according to the above recipe and compression mold. After compression molding in a machine, it was crushed in a crusher and sieved to obtain granules.

This granule 19 contains cis-(6)-shogaol 2
It contains 0.0m9, and adults should take 1-4g per day in several doses.

Example 6 ■ Crystalline cellulose 29 g ■ 10% hydroxypropyl cellulose ethanol solution 209 3 ounces of 6-ginger l Total 509 According to Moeki's recipe, ■ to ■ were mixed uniformly and made into a paste. After going into an extrusion granulator and granulating, the mixture was dried and sieved to obtain granules.

This granule NF contains cis-(6)-shogaol 2
It contains 0 mg, and adults should take 1 to 4 g per day in several doses.

Example 7' ■Corn starch 44.59■Light silicic anhydride 0.59 (3 ounces 6-ginger 5 total 509) Mix ■~■ uniformly according to the recipe in 4-, and make 200M
9 was filled into a No. 2 capsule.

This capsule! The capsule contains 20m9 of cis-(6)-shogaol, which can be administered for 1 to 1 day per day for adults. 1
Take the capsule in several doses.

Example 8 ■ Distilled water for injection Suitable insects ■ Budosuke 2001N9 ■ Cis-(
6)-Shogaol l Hg total m
After dissolving ① and ② in 5 mft of distilled water for injection, they were injected into an ampoule of 5- and autoclaved at 121°C for 15 minutes to obtain an injection.

Example 9 ■Sisu(6)-shogaol 0.05g■White petrolatum 259■Stearyl alcohol 229■White beeswax IF
M ■ Polyoxyethylene (25) Monostearate 2.39 ■ Sorbitan monopalmitate 2.79 ■ Propyl paraoxybenzoate 0.059 ■ Methyl paraoxybenzoate 0.059 9 + 32.85 total
100g According to the above recipe, ① to ② were mixed uniformly and dissolved by heating to obtain an ointment.

Claims (2)

[Claims] (1) The following formula ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ A novel cis-(6)-shogaol represented by (2) A therapeutic agent for keratosis containing a novel cis-(6)-shogaol as an active ingredient, which is represented by the following formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼.