JPS59486B2 - Anti-psoriatic active natural terpenes and their production method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to obtaining natural terpenes with anti-psoriasis activity by extraction from the rhizomes and leaves of various ferns.

Psoriasis and psoriasis are pathological symptoms that can appear on any part of the body and can affect the entire body, ie, from the head to the toes.

Therefore, the severity of these diseases is determined by the extent and depth of psoriasis progression that affects the skin layers. Although psoriasis is manifested on the skin, the dermatologist or general practitioner must not overlook the so-called complications, which are primarily manifested at the level of tissues such as joints, liver and kidney tissues. In advanced stages of the disease, psoriatic patients also show signs of diabetes, gout, and kidney failure, and the patient's condition worsens significantly. It must be noted that this is a recessive trait.
This is why the frequency of disease outbreaks among humans is increasing every month. In this regard, there is a report from the United Nations World Health Organization which estimates that clinically the number of people suffering from various forms of psoriasis can be estimated at 1% of the population. Although this problem has been known for some time, there have been virtually no therapeutic attempts to cure or ameliorate this pathological disease. Although certain cytostatic drugs have shown some efficacy in treatment, they are so toxic that patients cannot be treated more than once. In short, with the current recognition that psoriasis cannot be cured,
Only stopgap cures are available, which severely limits the lives of these patients in their daily work.

Dryopteriscrassirhizoma (DIyOpteriscrassirhizoma)
, Polypodium vulgare Linn (POlypOdiu
mvulgarelLinn), Polypodium leucotomos (POlypOdiumleucOtOmOs)
, Phlebodium
bOdiumdecumaunJ. Smith), Polypodium decmanum (POlypOdiumdec)
Cyath
eataiwaniana) and the rhizomes and leaves of Polydium aureum phosphorus (POlypOdiumau).
We have discovered that natural terpenes isolated from the rhizomes of P. reum Linn and POlypOdium triseriale have properties useful in the treatment of psoriasis and psoriasis patients of men and women and of all ages. I found it.

These terpenes include phanene and its isomers as well as other terpenes. A group of 152 patients suffering from psoriasis received a daily dose of 200T! 9 was treated orally, and symptoms disappeared in more than 80% of cases within 1 to 3 months. In a series of double-blind trials, Huanen was shown to be effective compared to a placebo of the same size and shape. Huanen is particularly useful for any type of psoriasis or psoriasis lesions, including the bulky palmar-plantar form of the disease. “Guidelines for Reproduction Studies for Safety Evaluation of Drugs for Humanity 1-S”
ductiOnStudiesfOrSafetyEv
aluatiOnOfDrUgsfOrHumanus
e”, Foot and Track Administration of the United States (FOOd)
andDrLlgAdministratiOnOft
According to studies carried out according to United States (HeUnited States) (1966), Huanen does not alter fertility in rats or mice and does not have any teratogenic effects in rats or mice.

When rats were orally administered 1η/kg of body weight daily for 20 weeks and observed at the end of 7 weeks with or without glass particles in their bladders, no carcinogenic activity was found.

The methods used in these studies were published in the British Journal of Cancer by Jull, J.W.
r), Vol. 5, p. 328 (1951), and the statistical evaluations used were published by ArcOs and co-workers in the Chemical Induction of Cancer.
r) 416 pages, Academic Press, Inc. (Academic Press) Nc)) 01-
3-K (1963).

The mechanism of action of Fanen appears to be related to membrane defects that apparently result in changes in the regulation of cell growth in the epidermis [Orfanos, C.I.
C. E. ) and Mahrle, G.
, DermatOlOgic, vol. 151, p. 1998215 (1976)] o vanen has no anti-inflammatory or anabolic steroid activity.

17/ Weight? It is essentially non-toxic.

Pure Huanen was previously known as Azeta, Etsuchi (Ageta
、． H. ) and their collaborators on a very small laboratory scale [Tetrahedron Letters 22, 1
pp. 442-50 (1963)].

In the method of the present invention, active terpenes can be extracted from the above-mentioned plant parts on an industrial scale in amounts used in pharmaceutical preparations. For optimal yields, leaves should be collected after the spores have developed. The method of the invention consists of the following steps.

The dried rhizomes are cut into strips of 2-3 cTn and the leaves are collected and dried.

Drying is carried out continuously by feeding the raw material into a rotating wire screen approximately 5 TrL wide and 25 m long which moves in a hot section at a temperature not higher than 80°C. By adjusting the speed of the rotating wire screen, about 0.5 tons of wet material can be dried in 1 hour. Granulation diameter up to 2 mm (ASTM sieve 10) and density approximately 0.3
The dry raw materials (residual moisture 5% or less) are granulated in a disc mill adjusted to produce particles of No. 6.

Extraction-Evaporation Extraction can be carried out using any solvent, but methanol, ethanol, n-hexane, dichloromethane, etc. are preferred, and methanol in a volume ratio of 1:4 is particularly preferred.

Purification 1. The semi-solid residue of the extract was dissolved in n-hexane/water (10:4).
) and after standing overnight in a separator, the salts and sugars are removed together with the aqueous phase.

2. To the n-hexane solution was added 1/10 of its volume of 10-20% KOH in ethanol, after stirring occasionally, 6
Leave it for a while.

The n-hexane phase is washed with water to neutralize it. 3. After evaporation, the residue is taken up in 3 times its volume of 96% ethanol and left at room temperature until complete precipitation, usually 48 hours.

4. The precipitate is filtered with suction through a glass filter of average porosity, washed with an appropriate amount of 96% ethanol until almost colorless, and dried under reduced pressure over KOH or concentrated sulfuric acid.

5. The precipitate was dissolved in n-hexane and incubated at room temperature of about 20°C for 1
A 0% by weight solution is obtained.

6. Add this solution depending on the amount of substance (500y to 1kg)
5 x 100 or 10 x 200 CTrLf) SI6O Geduran column (0.063-0.
2 mm, talc) and eluted with n-hexane.

Huanen elutes in the first 11 fractions (regarding 21). 7. The n-hexane solution is evaporated to a concentration of about 10% and left at room temperature to spontaneously precipitate crystals.

The crystals are filtered off with suction and the mother liquor is evaporated again to obtain a second mass of crystals. 8. Another method consists of using a neutral Al2O3 column (active) (5 x 100 cm) and eluting with n-hexane.

In this case, the eluate extracts only phanene. 9. Recrystallization is carried out in hot absolute ethanol.

The pharmaceutical compositions of this invention are formulated into conventional dosage forms by incorporating the phanene into a pharmaceutical carrier according to accepted pharmaceutical techniques.

The resulting pharmaceutical composition is also encompassed by the present invention. The active ingredient, ie, vanene, will be present in the compositions of the present invention in an amount sufficient to provide anti-psoriasis activity. Preferably, the compositions of the present invention contain from about 1 to 100 eta of active ingredient per unit dose.

Pharmaceutical carriers can be, for example, solid or liquid. Examples of solid carriers include lactose, magnesium stearate, Albaterra, sutucarose, talc, stearic acid, gelatin, agar, pectin, acacia, aerosil, and the like. Examples of liquid carriers include alcohol (eg, ethanol or propylene glycol), water containing a solubilizing agent such as polyethylene glycol, peanut oil, olive oil, and the like. The carrier or diluent can include a retarding agent such as glyceryl monostearate or glyceryl distearate, optionally with a wax. A wide range of pharmaceutical dosage forms can be used, and if a solid carrier is used, they can be formulated into tablets, placed in hard gelatin capsules in powder or granule form, or in the form of lozenges. . The amount of solid support varies over a wide range, but is preferably between about 25 and 250 η.
If a liquid carrier is used, the formulation may be a syrup, emulsion,
They can be made into soft gelatin capsules, suspensions or liquid solutions or sterile injectables for parenteral administration, such as ampoules. Pharmaceutical compositions of the invention that are suspensions or liquid solutions do not encompass mere suspensions or liquid solutions of the active ingredients in common solvents that are not suitable for internal use.

Dosage units in the form of tablets, capsules, lozenges, liquid suspensions or sterile injectables having anti-psoriasis activity, consisting of a sufficient amount of phanene and a pharmaceutical carrier to produce said activity, are also contemplated by the present invention. It is included.

Administration can be oral or parenteral, oral being preferred.

Preferably, the daily dose of active ingredient is about 100-600W9, even better about 50-300W1jf7. Advantageously, doses corresponding to 1 to 4 times per day are administered. When administered as described above, anti-psoriasis activity is obtained. Since psoriasis is a chronic, insidious disease with genetic etiology, it is preferable to use capsules or tablets to maintain stable blood levels, thereby allowing the drug to act continuously. be able to. Of course, when treating severe psoriasis caused by other drugs, such as steroids, or by the disease itself, intravenous injections or continuous infusions of serum can be used to avoid morbidity. It goes without saying that drops or emulsions can be used for children, and suppositories can be used for adults with damage to the upper gastrointestinal tract. Pharmaceutical compositions are manufactured by conventional means such as mixing, granulating, tabletting, if necessary, by dissolving or mixing with appropriate ingredients to form the desired composition. Next, the present invention will be explained with reference to Examples. In Examples 1 to 4, Polypodium leucotomos was used as a raw material, but all of the other plants mentioned above can be treated in the same manner to obtain phanene. Example 1 96% ethanol 10f at 70℃! and a black 10 sieve (2.
5mn) Extract the granulated dry drug 12K9 of Polypodium leucotomos for 1.5 hours.

Filter the extract and wash the entire filter with 96% ethanol 10e. ▲The filtered extract was heated under moderate reduced pressure (20~
Evaporate to dryness at 30 mmHg). The residue was diluted with n-hexane/
Take up in a mixture of water (15:101) and leave overnight in a separator.

The aqueous phase is removed and the hexane phase is clarified by adding 90% KOH in ethanol (10%) (51). The hexane phase thus clarified is washed with water and evaporated to dryness. The residue is dissolved in 500 ml of 96% ethanol with heating and the solution is left for 48 hours. ▲Filter and dry (yield approx.
07). The dried precipitate was dissolved in n-hexane 11 and SI-
60 Geduran column (100 x 1
0c7rL) and n-hexane 1
Elute with 1 and 2.

The first 11 fractions are evaporated and the residue is crystallized from hot hexane, benzene or absolute ethanol to give phanene (yield 127).

The physical properties of the obtained phanene are as follows. Melting point: 170-171°C Specific rotation: 16.51 (16°C, 0.79W9/chloroform 1CC) IR: 2830-2930 (1-1 (
methyl and methylene), 1455c:m-1 (ethylene), 13750rn-1 (inpropyl) NMR
(in CCl4): 5.25 ppm (olefin) UV:
198.6 mμ (1473) Fernen can also be obtained from the other plants mentioned above by the same operation as in Example 1.

The yield of phanene using the same weight of raw material as in Example 1 is listed below: Chacea. Taiwaniana・
・・・・・・・・・1.2～1.5t Polypodium decmanum...1″2～13y Polypodium triceria...13～13.57 Lepolipodium fulgare...2.28 ~ 2.57 Rhindriopteris cratsusi...2 ~ 37 Rhizomaphlebodium decumunun...12~137 Zei Smith Polypodium aureum...48~50t Phosphorus Example 2 Disk Dried whole plant of Polypodium leucotomos 36k passed through 10 sieves (2.5m0) and granulated in mill
9 was extracted successively with n-hexane at room temperature, the hexane solution was then evaporated to about 101, and K
90% ethanol solution (10%) of OH 1f! Add it and leave it overnight to make it clear.

The hexane phase is washed neutral with water and evaporated to dryness. Take up the residue in enough hot benzene to make a saturated solution (approximately 11)
. After cooling, the precipitate is filtered and crystallized in hot benzene or hot absolute ethanol. The crystalline material was dissolved in n-hexane (50
0 ml) and chromatographed on a 50 x 50 CTILSI-60 Gesillane column (Mercou d'Armushitat), eluting with 0.51 increments. The two first 0.51 fractions of the eluate are evaporated and the phanene is crystallized with hot hexane, hot benzene or hot absolute ethanol. (Yield 387). By the same operation as in Example 2, phanene can also be obtained from the other plants mentioned above.

The yield of Huanen using the same weight of raw material as in Example 2 is listed below: Example 3 Dry rhizomes of Polypodium leucotomos granulated in a disc mill and passed through a black 10 sieve (2.5 mm) 12kg
is extracted successively with dichloromethane at room temperature.

Dichloromethane was evaporated to dryness, and the residue was diluted with n-hexane 51
Take it. A hexane solution of 10% in 90% ethanol overnight
% KOHlI is added to clarify, then the hexane phase is neutralized by washing with water and evaporated.

The residue is taken up in n-hexane and chromatographed on a silica gel -60 column (100 x 10 cwL) using n-hexane.

The two initial eluate fractions are evaporated and the residue is crystallized from hot n-hexane, hot benzene or hot ethanol to give phanene (yield 257). By the same operation as in Example 3, phanene can also be obtained from the other plants mentioned above.

The yield of Huanen using the same weight of raw material as in Example 3 is listed below: Example 4 A whole plant of Polypodium leucotomos 12k9 (particle size measurement 2.5) granulated and dried by refluxing for 1 hour.
mT! L or black 10f) sieve) at 70°C with methanol 1
Extract with 00e.

The extract is filtered and the entire filter is washed with methanol 201. The methanol extract is evaporated and the residue is taken up in a mixture of n-hexane/water (3:2) (i.e. 301 and 20') and left in a separator overnight. The aqueous phase is removed and the hexane phase is clarified with 20% KOHlOI in 90% ethanol, washed with water, neutralized and evaporated to dryness. Heat the residue to 96%
Dissolve in ethanol 17 and leave at room temperature for 48 hours.

The precipitate is filtered and dried under reduced pressure over KOH.

The dried precipitate was dissolved in n-hexane 11 and neutral Al2O
3 (50 x 5 CTfL) eluting with n-hexane. Evaporate the first 51 and leave the substance n
- Crystallization from hexane, cyclohexane, hot benzene and hot anhydrous ethanol to obtain phanene (yield 12~
137).

By the same operation as in Example 4, phanene can also be obtained from the other plants mentioned above.

The yields of phanene using the same weight of raw materials as in Example 4 are listed below: isn't it.

Example 5 The ingredients are mixed and sieved into hard gelatin capsules.

Capsules prepared as described above are administered three times per day. Example 6 Mix phanene and sutucarose and granulate with 10% gelatin solution.

The granules are sieved, dried and mixed with starch, talc and stearic acid. These are sieved and made into tablets. Example 7 Ingredients are mixed and placed into soft gelatin capsules.

Claims (1)

[Claims] 1. Dryopteris crassirhizoma
), Polypodium vulgare Linn (Polypodium
um vulgare) Linn), Polypodium leucotomos (Polypodium leucotom)
os), Phlebodium decumunun zei-smith (P
hlebodium decumanum J. Smit
h), Polypodium decmanum (Polypodium
m decumanum), Chiacea taiwaniana (
Cyathea taiwaniana), Polypodium ovaleum Linn (Cyathea taiwaniana)
um Linn) or Polypodium triseriale (
Polypodium triseriale) is extracted with a solvent selected from the group consisting of methanol, ethanol, n-hexane and dichloromethane, concentrating the extract, taking up the concentrated residue in n-hexane/water, and recovering phanene from the n-hexane layer. Anti-psoriasis characterized by (
A method for producing tafanen with activity (including psoriasis). 2. The manufacturing method according to item 1, in which the rhizome of a plant is extracted. 3. The manufacturing method according to item 1, which involves extracting plant leaves. 4. Dry the rhizomes or leaves at a temperature not higher than 70°C to a residual moisture content of 5% or less, and granulate this to a density of approximately 0.5%.
36. The manufacturing method according to any one of items 1 to 3, characterized in that particles having a diameter of 2 mm or less are obtained and extracted. 5 The concentrated residue was mixed with (i) n-hexane/water (10:4)
and leave it in a separator overnight to separate the aqueous phase, (ii
) Add 1/10th volume of 10-20% KOH in ethanol to the n-hexane solution, stir occasionally, leave for 6 hours, wash with water to neutralize, and (iii) evaporate the solvent to remove the residue. (iv) suction filter the precipitate through an average porosity glass filter and collect enough 96% ethanol to become almost colorless; washed with ethanol and dried under reduced pressure over KOH or concentrated sulfuric acid; (v) the precipitate was dissolved in n-hexane to obtain a 10% by weight solution at room temperature of about 20°C;
vi) Transfer the solution to SI-60 Geduran
Chromatography in a column (0.063-0.2 mm, talc) or a neutral Al_2O_3 column (active), eluting with n-hexane, (vii) evaporating to a concentration of about 10% and n-hexane solution 2. Process according to claim 1, characterized in that the crystals are crystallized at room temperature, the crystals are filtered with suction, the mother liquor is evaporated again to obtain a second crystal mass, and (viii) recrystallized from hot absolute ethanol.