FI64892B - FRAMEWORK FOR ANTI-SORGHY EFFECTS - Google Patents

Description

IVfcSftr ·] [»] {11) ADVERTISEMENT zr / oqn lJ 1 'utlAggningsskrift 04öy ^ C ¢ 45) Γti- ·· ^ ^ -3 (51) K» .ik.3 / Int.ci.3 A 61 K 35 / 78, C 07 C 13/62 FINLAND — FINLAND (H) Patent application - Patentans6knln | 7 916l 6 (22) Hakemlspilvl - Anseknlnfadac 22.05.79 (23) Alkupdvl - Glltlfhetsdag 22.05 * 79 (41) Tullut (ulkbekal - Bllvlt offentllf 25.11.79

Patent and Registry Administrators ........, „. ,. ....

_ „_. . . . , (44) NlhUviksIpenon | s HEARING LAST DATE. - 31.10.83

Patent * och registerstyrelsen 'Ansttkan uthgd oeh utl.ikrlften publkend J

(32) (33) (31) Pyy ^ * «y privilege —Bejlrd prlorltet 24.05 * 78

Spain-Spain (ES) 470204 Proven

Styrkt (71) Conrad Limited, The Bank House, Frederick Street, Nassau,

Bahamas-Bahamasöarna (BS) (72) Francisco Ramon Alvarado Salgado, Tegucigalpa,

Blanca Zoila Ney de Alvarado, Tegucigalpa, Honduras (HN),

The present invention relates to a process for the preparation of an antipsoriatic natural terpene mixture by extracting various ferns from ferns. Antonio Horvath Sumi, Santiago, Chile (CL) (74) Oy Kolster Ab (54) Process for the preparation of an antipsoriatic terpene mixture - Förfarande för framställning av en terpenblandning med antpsoriatisk effekt .

Psoriasis and parapsoriasis are diseases whose symptoms can occur on any part of the body's surface. The severity of these diseases depends on the extent of the psoriatic effect on the skin as well as the depth in the skin layers. Psoriasis is also associated with complications that occur primarily at the joints and in tissues such as liver and kidney. As a result, patients with psoriasis may develop diabetes, gout, and even renal failure at an advanced stage, with significant deterioration in the patient's condition.

Psoriasis is a recessively inherited disease that explains the constant increase in the incidence of the disease. The World Health Organization has estimated that people with various clinical forms of psoriasis make up 1% of the population.

2 64892

Therapeutic attempts to cure this pathological disease have virtually failed. Certain anti-cell growth drugs have had some effect but are very toxic and cannot be treated more than twice by a patient due to their toxicity. Until now, psoriasis has only been alleviated and it has been taken for granted that psoriasis cannot be cured.

It has now been found that the natural terpenes isolated from the roots and leaves of the following plants Dryopteris crassirhizoma, Polypodium vulgara, Linn, Polypodium leucotomos, Phlebodium decumanum J.Smith, Polypodium decumanum, Cyatea taiwaniana and the plants Polypodium Aureum Linn and Polypodistaium are in the treatment of psoriasis and parapsoriasis of both sexes and all ages. These terpenes include e.g. fernene and its isomers. A group of 152 patients with psoriasis was treated with an oral daily dose of 200 mg, and this resulted in more than 80% of cases disappearing within 1 to 3 months. In a series of blinded tests, fernen proved to be effective compared with placebo (an ineffective medicine) of the same size and shape. Fernene is particularly useful in all types of psoriatic or Paraporiasis injuries, including the pustular palmarplantar form of this disease.

The present invention relates to a process for preparing an antipsoriatic terpene mixture by extracting the roots and leaves of the plants Dryopteris crassirhizoma, Polypodium vulgara, Linn, Polypodium leucotomos, Phlebodium decumanum J. Smith, Polypodium decumanum and Cyathea taiwaniana and Polypodium Aureum Linn and Linn that (a) the roots are cut into strips of 2 to 3 cm and the root strips and leaves are subjected to a continuous drying process on a rotating wire mesh at a temperature not exceeding 70 ° C; (b) the dried material with a residual moisture content of less than 5% is granulated in a plate grinder. is about 0.36, c) this material is extracted as a solvent having a dielectric constant of 1.890-9.08.

64892 d) the solvent is evaporated e) the semi-solid residue is taken up in n-hexane and n-hexane / water and allowed to stand overnight in a separator, and the aqueous phase, if any, is separated, f) 10-20% KOH is added to the n-hexane solution with occasional stirring. ethanol solution, one tenth of the volume of n-hexane soluble, and the solution is allowed to stand for 6 hours and neutralized by washing with water, g) the solvent is evaporated and the residue is taken up in three volumes of 96% ethanol and allowed to stand until complete precipitation has taken place, h ) the precipitate is filtered through a glass filter of conventional porosity, using suction and washed with a sufficient quantity of 96% ethanol until almost colorless, and dried in vacuo with KOH or concentrated sulfuric acid, i) the precipitate is dissolved in n-hexane to give a solution of about 20 ° C is 10% by weight, k) the solution is chromatographed on an SI-60-Geduran column (0.063-0.2 mm; Merck) or on a neutral N 2 O 2 column and a is eluted with n-hexane11a, l) evaporated to a concentration of about 10% and preferably crystallized from n-hexane, at room temperature and the crystals are filtered off with suction and the mother liquor is evaporated to give a second crystalline mass and m) the resulting crystals are recrystallized, preferably from hot, absolute ethanol.

Studies have shown that fernene does not affect fertility in rats or mice and does not have a malformative effect in rats and mice ("Guidelines for Reproduction Studies for Safety Evaluation of Drugs for Human Use" by the Food and Drug Administration of the United States (1966)).

Fernene did not show carcinogenic (carcinogenic) activity in rats tested at a daily oral dose of 1 mg / kg body weight for 20 weeks and in which the rat bladder was examined after 7 weeks. This research method is described in Jull, J.W.

Brit. J. Cancer; 5, 328 (1951) and the statistical evaluation used is described in Arcos and Collaborators, Chemical Induction of Cancer, p. 416, Academic Press Inc. New York (1963).

64892

The mechanism of action of fernen is apparently related to changes in the cell membrane that regulates skin cell growth (Orfanos, C.E. and Mahrle G. Dermatologic 151, 199-215, 1976).

Fernene has no anti-inflammatory or anabolic steroidal effect. It is essentially non-toxic up to a dose of 1 g / kg body weight.

Pure fernene has previously been isolated in very small amounts on a laboratory scale (Tetrahedron Letters No. 22, 1442-50; 1963).

The process according to the invention makes it possible to extract active terpenes from the roots and leaves of plants in industrial yields. To achieve optimal yield, the leaves should be harvested after the spore colonies have already developed.

The method according to the invention is described in more detail below.

Drainage

The roots are cut into 2-3 cm strips, the leaves are dried as they are assembled. Drying takes place continuously and the material is fed into a rotating metal mesh about 5 m wide and 25 m long, which is moved inside the hot compartment at a temperature of up to 70 ° C ». By appropriately adjusting the speed of the rotating metal mesh, about half a ton of moist material can be dried per hour.

Rakelstus

The dry material (residual moisture less than 5%) is granulated in a plate mill adjusted to a particle diameter of no more than 2 mm (ASTM sieve 10) at a density of about 0.36.

extraction

The extraction is performed as a solvent (dielectric constant 1, 890 to 9.08).

Purification 1. The oily solid residue obtained after evaporation of the extract is taken up in n-hexane or n-hexane / water and allowed to stand overnight in a separator, after which the salts and sugars are eliminated with any aqueous phase.

2. To the n-hexane solution is added one tenth of its volume of 10-20% KOH in ethanol, with occasional stirring, after which it is allowed to stand for 6 hours and the n-hexane phase is neutralized by washing with water.

3. After evaporation, the residue is taken up in three volumes of 96% ethanol and allowed to stand until complete precipitation has taken place, normally for 48 hours at room temperature.

4. Filter the precipitate through a glass filter of normal porosity using suction, wash with appropriate amounts of 96% ethanol until almost colorless, and dry in vacuo over KOH or concentrated sulfuric acid.

5. Dissolve the precipitate in n-hexane to give a 10% w / w solution at about 20 ° C.

6. Chromatograph the solution on an SI-60-Geduran column (0.063-0.2 mm, Merck) with a dimension of 5 x 100 or 10 x 200 cm, depending on the amount of substance (500 g to 1 kg) eluting with n-hexane. Fernene elutes into the first 1 liter fraction (a total of 2 liters are taken) Alternatively, a neutral N 2 O 2 column (5 x 100 cm) can be used, whereby only Fernene is extracted upon elution with n-hexane.

7. After evaporation to a concentration of about 10%, the substance is allowed to crystallize spontaneously, preferably from n-hexane, at room temperature. The crystals are filtered under suction and the mother liquors are evaporated to give a second crystalline mass.

8. The recrystallization is preferably performed from hot absolute ethanol.

The following example illustrates the invention

Example 1

12 kg of dry leaves granulated in a screen with NolO (2.0 mm) were extracted with 96% ethanol at 70 ° C

1.5 hours. The extract was filtered and washed with 10 liters of 96% ethanol on a filter. The filtered extract was evaporated to dryness under moderate vacuum (20-30 mmHg).

The residue was taken up in n-hexane / water (15:10 L) and allowed to stand overnight in a separator. The aqueous phase was eliminated and the hexane phase was clarified with a 10% KOH solution in ethanol (5 liters). The hexane phase thus clarified was neutralized by washing with water and evaporated to dryness.

6 64892 The residue was dissolved in 500 ml of 96% ethanol while heating and the solution was allowed to stand for 48 hours. It was filtered and dried (yield about 50 g).

The dry precipitate was dissolved in 1 liter of n-hexane and chromatographed on an SI-60-Geduran column (100 x 10 cm), eluting with 1 liter portions of n-hexane.

The first 1 liter fraction was evaporated and the residue was crystallized from hot hexane, benzene or absolute ethanol (yield 12 g).

Claims (1)

7 64892 Claim A process for preparing an antipsoriatic terpene mixture by extracting the roots and leaves of Dryopteris crassirhizoma, Polypodium vulgara, Linn, Polypodium leucotomos, Phlebodium decumanum J. Smith, Polypodium decumanum and Cyathea taiwanina, Linnum that (a) the roots are cut into strips of 2 to 3 cm and the root strips and leaves are subjected to a continuous drying process on a rotating wire mesh at a temperature not exceeding 70 ° C; (b) the dried material with a residual moisture content of less than 5% is granulated to obtain particles; with a maximum diameter of 2 mm and a density of about 0.36, c) this material is extracted as a solvent with a dielectric constant of 1.890 to 9.08, d) the solvent is evaporated e) the semi-solid residue is taken up in n-hexane or n-hexane / water mixture and allow to stand overnight in a separator, separating any aqueous phase, f) n-he a 10-20% solution of KOH in ethanol, one tenth of the volume of the n-hexane solution, is added to the xane solution with occasional stirring and the solution is allowed to stand for 6 hours and neutralized by washing with water, g) the solvent is evaporated and the residue is taken up three times (h) filter the precipitate through a glass filter of normal porosity using suction and wash with a sufficient quantity of 96% ethanol until it is almost colorless and dry in vacuo with KOH or concentrated sulfuric acid, (i) dissolve the precipitate; in n-hexane to give a solution of 10% by weight at about 20 ° C, k) the solution is chromatographed on a SI-60-Geduran column (0.063-0.2 mm; Merck) or on a neutral N 2 O 2 column and eluted with n-hexane, 8 64892 l) is evaporated to a concentration of about 10% and crystallized, preferably from n-hexane, at room temperature and the crystals are filtered off with suction and the mother liquor is evaporated to give a second crystal mass, and m) the crystals are preferably recrystallized from hot, absolute ethanol. 9 64892 Reproduction of the genomic antipsoriatic effect of the genome of extracellular fungus due to dry matter, Dryopteris crassirhizoma, Polypodium vulgara, Linn, Polypodium leucotoma, Phlebodiuin decumanum J. Smith, Polypodium decuman (a) three-dimensionally rotating metal at a temperature of up to 70 ° C; (b) granular detergent materials having a temperature of up to 70 ° C; a reduction of less than 5% in the square meter, with a particle size of 2 mm and a stem diameter of about 0.36, c) an additional material with a dielectric constant of 1.890 - 9.08, ) avdunstar lösningsmedlet; (f) the volume of the n-hexane solution under a mixture of 10 to 20% ethanol in KOH, the volume of which is determined by the volume of the n-hexane solution, and from the solution of the 6-dimensional and neutralization of the genome , (g) the addition of a solution and the addition of a three-fold volume of 96% ethanol and a full volume of the feed to the skeleton; for a period of 96% ethanol, the mixture is stirred under vacuum, and the mixture is evacuated with KOH or concentrated butter, vikt -%; ig # k) chromatographic solution on an SI-60-Geduran column (0.063 - 0.2 mm; Merck) or a neutral Al 2 O 4 column and eluting with n-hexane,