CS223879B2 - Method of gaining natural terpenes with antipsoriatic effect - Google Patents

Abstract

A process for extracting antipsoriatic natural terpenes from rhyzomes and leaves of Dryopteris crassirhizoma, Polypodium vulgara Linn, Polypodium leucotomos, Phlebodium decumanun J. Smith, Polypodium decumanum, Cyathea taiwaniana and rhyzomes of Polypodium aureum Linn and Polypodium triseriale comprises solvent extraction of a dry comminuted mass of the plant matter, evaporating off the solvent and purifying the residue.

Description

© The invention relates to a method of obtaining přirozenýc ter ^{p h} en antipsoriatt ^oJ ^ ou účmnosH extraction of the rhizomes and leaves of various ferns.

Psoriasis and parapsoriasis are pathological symptoms that manifest themselves on 'any part of the body's surface and are able to attack it completely,' ie. 'from head to toes. This proves. whereas the severity of this disease is therefore dependent on both the extent and depth to which the psoriatic process attacks the skin layers. It is noticeable on the skin, but no dermatologist or practitioner ^'of him ^at that cease ^t and ^k Zvan ^é complication of psoriasis, which occur mainly at the joints, tissues such as liver and kidney tissues which are manifested by psoriatic patients when advanced disease, showing signs of diabetes, gout, as well as renal insufficiency and worsening significantly.

It is important to emphasize that there is a genetic recessive character transmission, vehce ^Importan lie ^ite ^ novelties which ^{characterized} svěUuje ^d Enna vzrtetajtá heights ^yt for ^people with ^KE ras ^{y. T} ETO facts relating to the World Health Organization, which show that the number of people suffering from psoriasis with a clear activity of c ^h oro ^{would be} in her early khnickýich forms ^would molilo by ^T retracts ^ADR e2 no one percent of the population.

Despite prior knowledge of this subject, therapeutic attempts at therapy or amelioration of the pathological symptoms of the disease have been virtually without result. Some cytostatic drugs have shown relative efficacy in treatment, but are highly toxic and patients cannot be treated more than twice for their toxicity.

In short, currently available only palliatives, which along with te ^ ^ contact with sharp that it is clear ^{that p} soríázu Nete treat these patients maintain a very limited life in their daily tasks.

It has now been found that natural terpenes isolated from rhizomes and leaves of Dryopteris crassirhizoma, Polypodium vulgaro, Linn, Polypodium leucotomos, Phlebodium decumanum J. Smith. Cyathea taiwaniana and rhizomes Polypodium aureum Linn and Polypodium triseriale, have properties useful in the treatment of psoriatic and parapsoriatic patients of both sexes and at any age. These include terpenes Fernan and its isomers, and also remain ter ^{y p} en. ^S to ^P INA ^{152 p} acient suffering from psoriasis were treated with daily oral doses of 200 mg, and 'it led to vyimzern c ^h orob ^y at more than ^{80%, PR} and ^P and ^dV b ^During the ^d ob ^y by ^d 1 ^d 3 months . ^In serum steppe experiments, fernen proved to be effective compared to placebo of the same size and shape. Fernene is particularly useful in any kind of psoriatic and parapsoriatic lesions, including the pustular palmarplantar form of the disease.

Fernen does not affect the fertility of rats or mice and does not have a teratogenic effect on rats and mice, according to studies conducted and the Guidelines for Reproduction Studies for Safety Assessment of the Drugs for Human Use of the Food and Drug Administration of the United States (1966).

Fernen did not show carcinogenic activity in studies performed in rats after daily oral administration of 1 mg / kg body weight for 20 weeks i and as observed at the end of a 7 week period with and without the introduction of glass particles into the bladder. The methods used in these studies are described by J. W. Jull, Brit. J. Cancer; 5, 328 (1951) and the statistical evaluation used is described by Arcos et al., Chemical Induction of Cancer, p. 416, Academic Press Inc, New York (1963).

The mechanism of fernen efficacy appears to be related to membrane damage that leads to altered control of cell growth in the epidermis [C. E. Orfanis, G. Mahrle, Dermatologic 151: 199-215 (1976)].

Fernen does not have anti-inflammatory and anabolic steroid activity. It is substantially atoxic up to 1 g / kg body weight.

Pure fernene was isolated previously by H. Ageiiou sp. [(Tetrahedron Letters No. 22, 1442-50 (1963)] in very small amounts and on a laboratory scale.

The method of obtaining natural terpenes having an antipsoriatic effect according to the invention consists essentially in extracting rhizomes and leaves. Dryopteris crassirhizoma, Polypodium vulgare Linn, Polypodium leucotomos, Phlebodium decumanum J. Smith, Polypodium decumanum, Cythea tai.waniana, and rhizomes Polypodium aureum Linn and Polypodium triseriale, using the following chronologically consecutive stages:

(a) cutting the rhizomes into 2 to 3 centimeters strips and drying the rhizomes strips. and sheets by a continuous process on a rotary metal screen at a temperature not exceeding 70 ° C,

(b) granulating dry material with a residual moisture content of less than 5% in a disc mill to produce particles up to 2 mm in diameter and having an approximate density of 0,36;

c) extraction with a non-polar solvent having a dielectric constant of from 1.890 to 9.08, such as n-hexane, n-pentane, n-heptane, diethyl ether, petroleum ether, kerosene or benzene, preferably petroleum ether,

d) evaporating the solvent e ^j cleaning residue.

According to the invention, it is advantageous to carry out the cleaning of the residue by a process which comprises the following chronologically consecutive working steps:

i) collecting semi-solid residue of the extract in n-hexane / water (10: 4), wherein etxrakt allowed to stand overnight in a separator .and the aqueous phase was separated, ii) adding one tenth of its volume of 10- to ^20:% solution potassium hydroxide in ethanol to the n-hexane solution while stirring occasionally, allowed to stand for 6 hours and neutralized by washing with water, (iii) evaporating the solvent and bringing the residue up to three times its volume of 96% ethanol, leaving the mixture to stand (iv) filtering the precipitate with medium porosity glass filters under suction, washing with sufficient 96% ethanol until the product is almost colorless, and drying under vacuum over potassium hydroxide or concentrated sulfuric acid,

(v) dissolving the precipitate in n-hexane to obtain a 10% by weight solution at room temperature at about 20 ° C; (vi) chromatographing the solution on a silicone column in the form of aluminum hydroxide with a particle size of 0.063 to 0.065 ° C; 0.2 mm or on a neutral alumina (active) column, eluting with n-hexane; (vii) evaporating up to a concentration of approximately 10%, allowing the n-hexane solution to crystallize at room temperature, filtering the crystals by aspiration and repeated evaporation of the mother liquors. lye to obtain a second crystalline fraction, viii) recrystallization from hot absolute ethanol.

By the process according to the invention it is possible to carry out the extraction with industrial yields of active terpenes from said parts of the plants in sufficient amounts which can be used in the preparation of pharmaceutical preparations. For optimum yield, leaf collection should be performed when sporangium has already been developed.

Next they will. in more detail. the individual steps of the process according to the invention are described.

Drying The rhizomes are cut into strips of 2 to 3 centimeters, the leaves are dried immediately after harvesting. The drying is carried out continuously, the material being fed onto a rotary metal sieve of a width of about 5 m and a length of 25 m. moving in a hot chamber at a temperature not exceeding 80 ° C. By appropriately adjusting the speed of the rotary metal screen, about half a ton of wet material per hour can be dried.

Granulation

The dry material (residual moisture less than 5%) is granulated in a disc mill set up to produce particles up to 2 mm in diameter (ASTM sieve 10) having a density of about 0.36.

Extraction - evaporation

The extraction can be carried out with any solvent (dielectric constant from 1.890 to 9.08), preferably with methanol, in a ratio of ¹ : 4 by volume.

Cleaning

Extract the first semi-solid residue was taken up with hexane ^ ^/ water (W 4) and STEM ^p res night separator salts and sugars removed with the aqueous phase.

2. n-hexanovámu solution was added a tenth ^h oo ^BJJ emu ¹⁰ - ² DEG ^hy droxidu% Solutions of potassium in ethanol and occasional mixing was allowed to stand for 6 hours. The n-hexane phase is neutralized by washing with water.

3. After evaporation the residue was taken up in trojiteso ^{BK SV} him to him ^b eteanolu ^98% and allowed to stand until precipitation was complete, usually 48 hours at room temperature.

The precipitate was filtered fourth glass filters poráznosti medium by suction, the variable ^y is ^d ostatečným amount of ^96% ethanol until it is almost colorless and dried under vacuum over KOH or concentrated sulfuric acid.

5th precipitate is dissolved in n-hexane until zfeká 10% of ^the mites ^(h possible.) At room teptote about 20 ° C.

6. The solution is chromatographed on a column of ^S I-60 Geduran ^(0.063 to ^0.2 mm, Merck), 5 x ¹⁰⁰ or ¹⁰ x ²⁰⁰ щ depending on materiæ solubilizing amount (500 g to 1 kg) eluting with n- hexane. Fernan is contained in the first one liter ^{and F} reaction (vzhtedem to htrům ^2).

7. After evaporation to a concentration of approximately ¹⁰ %, the n-hexane solution ^is removed for spontaneous crystallization at room temperature. The crystals are filtered by suction and the mother liquors are again evaporated to give a second crystalline fraction.

⁸ . Another method ^{of (p} o s ^{s CI} in ^{PPLICATION s} stoupců neutráhnho hhnitého oxide (aktívmho) (5 x 100 cm) and eluted with n-hexane. In this case the eluate is extracted only Fernan.

9. Recrystallization is carried out from hot absolute ethanol.

^Obtaining birth ^to a rearward Tyz t ^ ^f yzikálně alni and chemical characteristics:

a) ^b ^ Mek fine mikrokrystebcký

(b) white;

(c) tasteless,

(d) odorless

(e) not soluble in ethanol (cold) and in water,

g) molecules, wt. 410 392

(h) spec optical opacity aD = +1 (c = 0,5; in chloroform),

(i) UF absorption spelktrum;

j) melting point 170-171 ° C.

are prepared in conventional dosage forms by incorporating fernene into pharmaceutical carriers according to accepted pharmaceutical practice. The active ingredient, i.e. Fernan is present in the composition produced according VYN ^AL cut in and ^the amount dostatefoém vyvotela antipsoriatic activity.

The compositions prepared from the products made according to the invention contain, preferably 1 to 100 mg of active ingredient per dosage unit. For example, the pharmaceutical carrier may be a solid or a liquid. Examples of solid carriers are lactose, magnesium stearate, clay, sucrose, italek, stearic acid, gelatin, pectin, acacia, aerosols and il ^p o ^{d. P Lady} of manufacture hot nose al ^ICU is ^to shave (e.g. ethanol or propylonglykol), water containing a solubilizing agent such as polyethylene glycol, peanut oil, olive oil · like. The carrier or the diluent may include retaMtení ^C m ^d la. for example, glycerol monostearate or glyceryl distearate as such or with a wax.

W can be used ^; of the pharmaceutical form. For example, if a solid carrier is used, the preparation may be converted into tablets, placed in hard gelatine wafers in the form of powder or granules, or formed into lozenges.

The amount of solid carrier may vary within wide limits, but preferably from about 25 mg to 250 mg. Uses--! In the case of a liquid carrier, the preparation may take the form of a syrup, emulsion, soft, latin capsule, suspension or liquid solution or a sterile injectable form for parenteral use, for example in an ampoule.

The pharmaceutical compositions prepared from the producer of the ^kt disburse ^{y p} e of the ^dl vynátezu suspensions or liquid solutions do not include simple liquid suspensions or solutions of active ingredient in common unsuitable solvents for internal use to produce the desired ^E termakotogpckéto Mink him.

A unit dosage form in the form of a tablet, capsule, lozenge, liquid suspension or sterile injectable liquid for intramuscular administration with an antipsoriatic effect containing a pharmaceutical carrier and a phenol in an amount sufficient to produce such an effect can be prepared from the products of the present invention.

Administration can be oral or parenteral, preferably oral. The active component is preferably administered in a daily dosage amount of about 100 mg to 600 mg, and more preferably as · ^{i 50 d 3} 00 m ^g. It is more ODNs ^{h e p} from ^A vat equivalent doses 1 to 4 times per day ·. When applied as described above, antipsoriatic activity is achieved.

Because psoriasis is a chronic insidious process of genetic etiopathogenicity, it is preferable to use capsules or tablets to maintain stable blood concentrations, allowing continuous drug action.

^^ catch ^{when treated} vMné ^and bustle of ^y

The pharmaceutical preparations of the invention psoriasis induced by other drugs, such as in the case of steroids, or the disease itself, can be used by intravenous injection or continuous infusion to prevent the crisis.

: Should ^b y ^{t b} e noted also ^that children from zfejmých ^{d troduction} of ^p ou ^running in ^{k and} e ^{k p} or emulsions and adults, in cases with problems of the upper digestive tract, suppositories.

The pharmaceutical compositions are prepared by conventional techniques such as mixing, granular and Esováním, if before ^the TNE, or by mixing or dissolving the respective components for the desired composition.

The invention is illustrated by the following examples.

EXAMPLE 1 kg of dry leaves granulated through a sieve c. Ϋθ ^(2, 5 mM) with 10% -ethanol htry 96 ^pl with TE OTE ⁷⁰ ° C b about ^yl extracted after ^b u ^d about 1.5 hours. Exrakt was filtered and washed on fWru l ^y r ^{0 lit 96%} ethanol. Filtered extíakt ^would l evaporated ^to dryness under a moderate vacuum (20-30 mm Hg).

Zb ^y te l ^{to the} extract ^and a mixture of n ^monthly n-hexane / water ^{(15: 10} htrům) and the extract b ^yl allowed to stand overnight in a separator. The aqueous phase was removed to .a ^{h of} exanová ^yčiř ENA bath in ^{10% of} mites ^to em droxidu ^{hy d} raselného in 90% ethanol ⁽⁵ l ^y). Taltío in ^yčí REN ^and the hexane phase forms promýváitím ^was neutralized and water and evaporated to dryness.

^ZBY te ^{to be} extended ^p l u ^s t h ^e I n ^{500 96%} etoanolu under heating and the solution was allowed to stand for 48 hours. It was filtered and dried (yield: about 50 g).

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

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

EXAMPLE 2 ^{36 g} of dry ^treatment plant ce ^{y s g} ranulovan in a disc mill to pass a no. 10 (2.5 mm) were continuously extracted with n-hexane at room temperature, the hexane solution was concentrated to about 10 liters and clarification ^BC overnight in a separator HTree 1 ^{to 10%} by mites ^{hydroxyacetophenone d} u ^d s raseln ^{eh 9} ° ethanohi%. The hexane solution was neutralized by washing with water and then evaporated to dryness.

The residue was extracted with hot benzene, sufficient to give a saturated solution (about 1 liter). The precipitate was filtered through a glass and filtered from hot benzene or hot absolute ethanol.

Cr ^y stalický material ^was extended at ^{St p} en in n-hexane (500 mL) and to ^the chrrmalografován One ^l of ⁵ X 50 cm 0 SI- ⁶ ° Geduran (Merck- -Darmstadt), eluting with 500 ml dose. Two ^{e p} r ^{s 5} 00 m ^l fractions b ^y L ^y, evaporated and crystallized from hot. hexane, hot benzene or absolute ethanol (yield: 38 g).

P ad 3 Rikli ^{12 kg} suc ^h yc ^h oiMenků granulated in ^di aligns correctly with ^the m ⁱ yne ^ab Rosh ^{yp L} of sieve no. 10 ^(2.5 ιη: ^b ^ y ^l o ^h o spare ^and NO at dichtormethanem room temperature. Dichloromethane was about ^d pa en ^{d of} the drought and ZB tek ^y b ^y L ^y extracted with 5 l n ^h Exan.

The hexane solution was clarification overnight ffirem 1% W mites ^{to y} uh droxidu toaselného in ^9% ethanol and 0 ^h exanová phase B ^y la ^p Otom neutralized by washing with water and evaporated.

The residue was taken up in one liter of n-hexane and chromatographed on a silica gel column —60 (100 x 10 cm) with n-hexane. ^p r ^{s of} two ehiátové fraction b ^yls from ARENA ^{P y,} and the residue was crystallized from hot n-hexane, hot benzene or hot ethanohi (yield ^{25 g).}

Example 4

By heating under reflux. temperature for 1 hour was extracted with 12 kg of granulated and dried whole plant (granulate size of ^2.5 mm or ^Stiomi. 10) ^y 100 liters of methanol at 70 ° C. The extract was filtered and the filter washed with 20 L of methanol.

Methanolic extract was evaporated and ZBY ^t e ^k b ^{yl characterized} exempts into a mixture ^of n-hexane ^{/ d} and a ^{(3: 2) (i.e., 3} 0 ² 0 ltírům) b l ^{y p} onechán. stand overnight in separators. The aqueous phase was removed and the hexane phase was clarified 10 ^l i ^{t y} r 0 ^2% solution of H ^y droxidu to-asetoéto in ethanol was neutralized by washing with water and evaporated to dryness.

The residue was dissolved in 1. L of hot ^96% ethanol and ^p onechán state pn te ^P tot ^of room temperature for 48 hours.

The precipitate was filtered off and dried under vacuum over potassium hydroxide.

The dry precipitate was dissolved in 1 liter of n-hexane and chromatographed on neutral alumina (50 x 5 cm) eluting with n-hexane.

rvrnc ^{hour PE P} t b htrů ^yl evaporated and the material was crystallized from n-hexane, cyclohexane or hot benzene and hot ethanol (yield: 12 to 13 GJ.

The following formulation examples for Ar ^{and p} TU ^y n and ^l ^ ez only tíustruj limitative.

Example 1

Component Quantity Fernan 40 mg Lactose 60 mg ^l uj ^s mites ^to em · 10% želatmy. The granutes are sieved, dried and then blended with starch, talc and stearic acid. They are screened and converted into tablets.

Example 3

Component Quantity

The ingredients are mixed, sieved and formulated into hard gelatin wafers. Wafers at ^p Raven described from ^p Úsobí to soak · ^s · TRL times daily.

Example 2

Component Quantity Fernan 60 mg sucrose 25 mg starch 20 mg ^to 5 m tale, stearic acid 5mg Fernan ara & taken note oil microcapsules ¹⁰⁰ mg

These components are mixed and. form into soft gelatin capsules.

Fernen and sucrose are mixed and granulated

Claims (2)

First caused ^b ZIS ^{ka tio n s p p} ter řírodních .enů with antipsoriatic action, characterized in that the extracted rhizomes and leaves of Dryopteris crassirhizoma, Polypodium vulgare Linn, Polypodium leucotomos, Phlebodium decumanum J. Smith, Polypodium decumanum. Cythea taiwaniana and rhizomes Polypodium aureum Lin.n · and Polypodium 'triseriale, following the chronologically following steps: (a) cutting the rhizomes into strips of 2 to 3 centimeters and drying the rhizome strips and leaves by a continuous process on a rotary metal screen at a temperature not exceeding 70 ° C; b) granulating the dry material ZB Numeric ^{y s} n ^{lower level of} humidity than 5% ^Dis b ^s m mill to produce particles with a diameter of 2 mm and a density of approximately 0.36; c) extracting the non-polar solvent with a dielectric constant of from 1.890 to 9.08, e.g. n-hexane, n-pentane, n-heptane, ^di eth ^yl years ^H & rem ^p etrole ^{t Here,} I ^p etrotejem or benzene, preferably petroleum ether, (d) evaporation of the solvent; and e) cleaning the residue. ² . ^Zp Úsobí ^p o ^{d b} le ODU 1 načujto ^ t ^ ^nus that ^read ztytku óhronologicky comprises the following working steps: i] Assembly of the semi-solid residue of the extract in n-hexane / in ^D (10: 4 ^ and the extract is allowed to stand overnight in a separator and the aqueous phase was separated, ii) adding one-tenth its volume 10When 20% solution hydroxdu potassium of ethanol to n-hexanovému solution, thereby ob and ^no MIC was Ponec ^ st ^ate after at ^{d b} of ⁶ hours and neutralized by washing with water, iii) evaporating the solvent. introducing the residue into the .trojnásobku ^h oo ^b him eteanolu ^96%, while the mixture was allowed to stand ^for about nastane- ^d she ^le ^ r ^ en ^ iv) filtering the precipitate of the mean porosity glass filters under suction, washing with ^sufficient Nym ^{LeVeLs 96} % ethanol until the product is almost colorless, and vacuum dried over potassium hydroxide or concentrated sulfuric acid, v) dissolving the precipitate in n-hexane to ^OBTAINING motnostn ^{lh of a 10%} solution of PN room temperature about 20 ° C, vij CONTRARY TO chromatograíf solution through a column of silicone in the form of aluminum hydroxide of the grain size 0.063 to 0.2 mm, or at column ^E with neutral to ^y col ^IR mkem hhrntým active, eluting with n-hexane, vii) evaporation to a concentration přibhžně ¹⁰ Ponec ^h ARN n-hexanoic ^s solution through crystallisation ^l izova ^t at te ^p aunt room ^ ^f iltrace crystals by suction and re-evaporation ^of the materials ^No measurement NYC ^{hr of} liquor к gain ^{and d} m ^s ruh him crystalline portion, VIII) recrystallized from hot absolute - ethanol.