PL43326B1 - - Google Patents

Description

The invention relates to a method for obtaining previously unknown substances with psychotropic effects, psilocybin and psilocin, from certain species of mushrooms. The method according to the invention consists in extracting the active substances from Psilocybe mexicana Heim or from artificially cultivated mushroom material obtained from cultures or biological variants or mutants of this mushroom by inoculating natural or artificial media and maturing these cultures in the light or in the dark at a constant temperature between 18 and 27°C according to known methods and then purifying them, separating them from each other and optionally freeing them from halogen. Furthermore, it has been found that psilocybin and psilocin can be obtained in a pure state in accordance with the above-mentioned procedure, starting from mushroom material. *) The patent owner has declared that the inventor of the invention is Roger Cailleux. of natural origin of the genera Psilocybe, Stropharia, Conocybe, Panaeolus, Amanita or Russula, or from artificially cultivated mushroom material obtained from cultures of particular mushroom species or their biological variants or mutants. It is already known (R. Heim, Cr. hebd. Seances Acad. Sci. 242, 965 (1956)) that a number of mushroom species of natural origin from Mexico, Eastern Siberia, Kamchatka and other countries of Asia and America are used for ritual purposes or as stimulants or intoxicants. They produce specific effects on the body and, above all, on the human psyche. These so-called hallucinatory mushrooms include the following genera: Psilocybe, Stropharia, Panaeolus, Conocybe, Amanita, and Russula. However, to date, it has not been possible to isolate active substances from mushrooms of natural origin or from artificially cultivated mushroom material. Nor has it been possible to cultivate species in the laboratory, starting from mushrooms (with hallucinatory properties) of natural origin, such species that would produce active substances in quantities sufficient to obtain these substances on a preparative scale. The invention relates to a method by which previously unknown active substances from hallucinatory mushroom species can be obtained, primarily Psilocybe mexicana Heim, Psilocybe caerulescens Murrill var. Mazatecorum Heim, P. caerulescens Murrill var. nigripes Heim, P. Zapotecorum Heim, P. semperviva Heim and Cailleux, P. Aztecorum Heim, Stropharia cubensis Earle. These active substances can be obtained in pure form by known methods, either directly from fungal material of natural origin or from fungal material cultivated from cultures derived from natural or artificial substrates, whereby the cultivation of these cultures is carried out according to methods that would allow obtaining the active substances in technically useful quantities. The procedure is usually as follows: for cultivation on natural substrates, compost is prepared from fermented wheat straw, a mixture of corn leaves and stalks, and wild grass blades, which is then washed with plenty of running water, crushed in a clay bowl, and sterilized in an autoclave. The compost is inoculated with mycelium from the original cultures and allowed to mature for about two weeks at a temperature of 24-27°C, while the cultures are covered with sterile sand and left at a temperature of 18-27°C in glass boxes in daylight, while maintaining constant humidity. After four to five weeks, the fruiting bodies appear: harvesting occurs within one to two months of the start of spore formation. However, since obtaining larger quantities of starting material in this way is difficult, other cultivation methods have been developed. It was unexpectedly found that in vitro fungi grown on substrates rich in nutrient form, instead of mycelium with large quantities of fruiting bodies, almost exclusively active fungi and partially sclerotia, while on substrates poor in nutrient, they form the familiar fruiting bodies. For example, for agar medium (1.5% agar content), the optimum for pericarp formation is at a dry substance concentration of malt extract of 0.2-0.7%, while the optimum for mycelium and sclerotia formation is at a concentration of 4-10%, depending on the mushroom species. While daylight is necessary for pericarp formation, it has been found that during the maturation of cultures in the dark, mainly abundant sclerotia or mycelium formation occurs. The best yields of active fungal material (mycelium and sclerotia) are obtained when surface cultures are grown on a malt extract nutrient solution (beer wort or a commercial malt extract preparation). with a dry matter content of 4-10% and when these cultures are left to mature in the dark at a constant temperature between 22-26°C. For good growth, 0.2% agar should be added to the nutrient solution. In this almost liquid medium, the fungus finds sufficient support to quickly form a closed mycelium cover. The addition of Fe11 salt is essential, and additions of zinc, potassium, calcium and magnesium salts containing nitrate, phosphate and sulfate ions, as well as yeast extract or corn steep liquor, have been found to have a very beneficial effect. This cultivation method represents a great technical progress, as it allows for obtaining a more than ten-fold higher yield of active starting material in a shorter time and with less work than cultivation on natural substrates. The active fungal material (fruit bodies, sclerotia or mycelium) is carefully dried in a stream of air or in a vacuum at a temperature of 20-40°C, finely ground and thoroughly extracted with water, a lower alcohol Aliphatic or a mixture of water and a water-miscible organic solvent. Extraction is carried out at room temperature. The extracts are evaporated in a vacuum at low temperature, and the residue is degreased with petroleum ether and extracted with acetone or a chloroform-alcohol mixture to free it from inactive accompanying substances. The remaining ballast substances are separated by dissolving the residue in the smallest possible amount of water and reprecipitating with absolute ethanol or acetone. The filtrate is evaporated in a vacuum at low temperature. Further purification is carried out by chromatography on a column of cellulose powder. Elution is carried out with water-saturated butanol or another water-immiscible alcohol. The collected fractions are tested for active substance content with Keller's reagent (iron chloride in acetic acid and concentrated sulfuric acid). Fractions showing a positive color reaction are combined and, if necessary, separated again on a chromatographic column packed with cellulose powder. The faster-moving zone, which produces a pure blue color in the Keller reaction, is eluted from the chromatogram. This zone yields the active substance called "psilocin," while the slower-moving zone yields the second, quantitatively predominant active substance, "psilocybin," which produces a violet color reaction. The active substances obtained from the column in a relatively pure form contain halogen and therefore do not crystallize. Only through chemical treatment can the halogen be removed, yielding compounds capable of crystallization. To achieve this, an aqueous solution of the active substances is treated with silver charcoal, and the filtrate is freed from excess silver ions using hydrogen sulfide. The filtrate is then concentrated in a vacuum at low temperature, with the active substances crystallizing from the concentrated solution. For analytical purposes, psilocybin is recrystallized from methanol or water. Fine, snow-white needles are obtained from water, and colorless hexagonal plates or prisms containing methanol melt with decomposition at 195-200°C. The resulting compound dissolves in 120 parts by volume of boiling methanol or 20 parts by volume of boiling water; in higher alcohols or other organic solvents, it is very sparingly soluble or completely insoluble. For analysis, the pure substance is dried in a vacuum at 100°C, with a weight loss of 10.4%. The results of elemental analysis correspond to the molecular formula Cs2Hl7O4N2P (molecular weight 284.2). Psilocybin is an amphoteric, optically inactive compound and dissolves readily, forming salts, in dilute aqueous solutions of mineral acids and alkalis. A solution of psilocybin in 80% aqueous ethanol reacts weakly acidic (pH = 5.2). The ultraviolet spectrum in methanolic solution shows maxima at 222, 267, and 290 µL. For analysis, psilocybin is purified by rechromatography on a column packed with cellulose powder using water-saturated butanol or by treatment with sodium bicarbonate in aqueous solution and shaking with ether or another organic solvent. The results of elemental analysis correspond to the molecular formula Cl2H16NO2. Psilocin crystallizes from methanol or acetone: it is moderately soluble in water and readily soluble in dilute acid. Melting point 173-176°C (decomposition). Psilocin is characterized by an ultraviolet spectrum (in methanol solution) with maxima at 222, 260, 267, 283, and 293 mp, and a Keller color reaction in which, unlike psilocybin, it gives a pure blue color. When injected subcutaneously or administered orally, 2-8 mg of psilocybin produces in humans an outwardly manifested sense of well-being, accompanied by a lack of excitability and apathy. At higher doses, certain noticeable external changes occur in addition to vegetative symptoms. The compounds discussed can be used to study mental illnesses and psychoses of various origins and can be used as an effective adjunct in the psychotherapeutic treatment of the mentally ill. Example 1. Psilocybin and psilocin from Psilocybe mexicana Heim of natural origin. For cultivation on a natural substrate, compost is prepared from fermented wheat straw. The compost is washed with plenty of running water, ground in a clay bowl, sterilized in an autoclave, and then inoculated with mycelium from the original cultures and allowed to mature for approximately two weeks at a temperature of 24-27°C. After this time, the cultures are covered with sterile sand and left in glass boxes in daylight at a temperature of 21-22°C. After 4 to 5 weeks, fruiting occurs: harvesting occurs within one to two months from the time spores begin to form. - 3 - Ripe fruiting bodies of Psylocybe mexicana * Heim are then carefully dried in an air stream at 30°C. 54 g of the dried mushrooms are finely ground and shaken once with 600 ml of methanol and three times with 300 ml of methanol for 1/2 hour. The extracts are purified and evaporated to dryness in a vacuum. Residue: 12 g. The methanol extraction residue is triturated four times with 250 ml of petroleum ether and another two times with 100 ml of chloroform with 10% alcohol added to remove fats. The still undissolved residue is dissolved in 10 ml of water and slowly added with 100 ml of absolute alcohol, enriching the solution in active substance. This operation is repeated two or three more times. The decanted solutions are evaporated in vacuo to dryness, redissolved in methanol, and treated with 20 g of cellulose powder with 5% water added. The methanol is evaporated again in vacuo, and the cellulose powder loaded with the active substance is introduced into a column filled with 100 g of powdered cellulose with 5% water added (the column is previously purified by washing with water-saturated butanol). Elution is carried out with water-saturated butanol, and 20 ml fractions are collected. The residue after evaporation of the individual fractions is tested for active substance content using Keller's color reaction. For this purpose, 0.25 mg of the sample from the evaporation residue is treated with 2 ml of Keller's reagent. The fractions with a positive color reaction are combined. The amorphous pyrolysis product is dissolved in 20 ml of water and shaken with 0.5 g of silver carbonate. After filtration, silver ions are removed from the solution with hydrogen sulfide, and the solution is then carefully concentrated. Psilocybin pxl crystallizes from the concentrated solution as colorless, fine needles (yield: 200 mg). Psilocybin is obtained from the pericarp only in trace amounts. Psilocybin is obtained in an analytically pure state after repeated recrystallization from methanol or water. It is dissolved in 120 parts by weight of boiling methanol or 20 parts by weight of water at boiling temperature. Colorless prisms are obtained from methanol, which melt at 195-220°C with simultaneous decomposition. The given values correspond, according to analysis, to the molecular formula C1aH2Oa2P2. The ultraviolet spectrum obtained in methanol shows the following maxima: 222 m[i], 267 m[j, and 290 m[i]. The new active substance has an amphoteric character. It dissolves in both dilute aqueous solutions of alkalis and aqueous solutions of acids. A solution of psilocybin in 80% aqueous alcohol reacts acidically (pH 5.2). Example II. Obtaining psilocybin and psilocin from Stropharia cubensis Earle of natural origin. Fruit bodies of Stropharia cubensis Earle collected in Mexico are air-dried in the shade at room temperature. 24.2 g of the dried pericarp are finely ground and shaken at room temperature, once with 300 ml and three times with 150 ml of methanol for 1/2 hour. The extracts are combined and evaporated in a vacuum to dryness. The residue (6 g) is de-fatted by trituration four times with 125 ml of petroleum ether and then three times with 50 ml of chlorofluorocarbon containing 10% ethanol. The remaining undissolved residue, approximately 5 g, is dissolved in 5 ml of water, and other accompanying substances are precipitated from this solution by slowly adding 50 ml of absolute ethanol, thereby enriching the solution in active substances. The purification is repeated two or three more times. The decanted solutions are combined and evaporated in vacuo to dryness. The residue is taken up in methanol and treated with 10 g of cellulose powder containing 5% water. The methanol is evaporated in vacuo, and the cellulose powder containing the active substance is introduced into a column packed with 50 g of cellulose powder containing 5% water (the column is previously washed with water-saturated butanol). Elution is carried out with water-saturated butanol, and 10 ml fractions are collected. Individual fractions are evaporated in a high vacuum at a bath temperature of no more than 50°C and tested for active substance content using glacial acetic acid containing ferric chloride and concentrated sulfurous acid. For this purpose, samples of the 8.85-milligram residue are treated with 2 cm³ of Keller's reagent. Active fractions give a violet (psilocybin) or pure blue (psilocin) color in the Keller reaction. Fractions giving a positive color reaction of the same color are combined. The amorphous powder obtained as the residue after evaporation of the above fractions is dissolved in 2 ml of water and shaken with 0.25 g of silver carbonate. After filtration, excess silver ions are removed with hydrogen sulfide. Psilocybin crystallizes from the solution concentrated under conservative conditions in the form of colorless, fine needles. 58 g of crystalline psilocybin are obtained, corresponding to a yield of 0.24% based on the dried pericarp, and traces of psilocin. Example III. Obtaining psilocybin and psilocin from pure cultures of Psilocybe semperviva Heim et Cailleux. a. Obtaining the vaccine: Pure cultures of spores of the Mexican cap mushroom Psilocybe semperviva Heim et Cailleux are introduced onto a medium of beer wort and agar. For this purpose, spores falling from the gills of the mature pericarp are captured on a sterile plate, placed on beer wort with agar, and allowed to mature. Inoculation material is obtained from the primary cultures thus prepared as follows: The mycelia are scraped off with a sharp spatula under a sterile water supply, creating a suspension containing fine mycelial flocs. This suspension is then inoculated into Erlenmeyer flasks containing the nutrient medium and saddle-shaped porcelain fillings. Experiments are best performed in 300 ml Erlenmeyer flasks containing 50 g of a saddle-shaped filling (size 1 cm, weight approximately 0.9 g) and 80 ml of a nutrient medium consisting of beer wort with approximately 4% dry matter and 0.2% agar. The culture matures at 24°C; after two weeks, a dense mycelium cover forms on the surface. The entire culture is shaken for 30 minutes on a rotary shaker, with the sharp edges of the saddle-shaped filling grinding the mycelium. This produces a fine suspension of mycelial flocs. The inoculum thus obtained is sufficient to inoculate 50 liters of medium. b. Obtaining the culture: Fresh light beer wort without added hops is diluted with water to 8% dry substance. One liter of this solution contains: FeSO4 . 7H20 0.00417 g ZnSC4 . 7H20 0.00172 g Ca(NO3)2 1.0 g KI12P04 0.0624 g MgSO4 . 7H20 0.0624 g KCl 0.0312 g Agar-Agar 2.0 g This medium is poured in 1 liter portions into penicillin flasks and sterilized in an autoclave for 25 minutes at 108°C. After cooling, the contents of the flasks are inoculated with 2 ml of P. semperviva Heim et Cailleux suspension. The resulting cultures are matured in the dark at 24-26°C. A mycelial cover is formed as described under IIIa. c. Isolation of the fungal material: After seven weeks, the mature culture is filtered through gauze, the fungal material is pressed out and dried in a vacuum at 30°C. From one batch of 100 penicillin flasks containing 100 liters of medium, 2540 g of dry material is obtained when harvested after 49 weeks, i.e., 25.4 g per liter of cultured medium. The culture filtrate, which also contains a certain amount of the active substances, is treated in the same manner as the fungal material described below. d. Obtaining active substances: 306 g of dried fungal material is thoroughly pulverized and shaken three times with 500 ml of chloroform and then three times with 500 ml of chloroform containing 10% ethanol. Inactive accompanying substances are dissolved in the amount of 2.8 g. The pre-extracted fungal material is thoroughly leached once with 3 liters and three times with 1.5 liters of methanol. The combined extracts are evaporated under reduced pressure to dryness, leaving a light brown precipitate of 17.5 g. To separate it from fatty impurities, this precipitate is dissolved in 17.5 ml of water and the suspension is shaken once with 500 ml and twice with 250 ml of petroleum ether. The petroleum ether solution contains 0.75 g of inactive accompanying substances. The remaining aqueous solution is concentrated in vacuo to about 25 ml and, with vigorous stirring, added 250 ml of absolute ethanol. The solution containing the active substances is separated by decantation from the sticky precipitate that forms. The precipitate is redissolved in a small amount of water and treated with a tenfold volume of absolute ethyl alcohol. This purification of the precipitate by precipitation is repeated twice more. The solutions are combined and evaporated in vacuo to dryness. 11.7 g of residue is obtained, which contains the total amount of active substances. For chromatography on a cellulose column, the residue is dissolved in the smallest possible amount of 50% methanol, and the resulting solution is thoroughly mixed with 40 g of cellulose powder and dried in vacuo. The cellulose powder thus loaded with the substance is introduced into a cellulose column, the packing of which was prepared by desliming 50 g of cellulose powder with water-saturated butanol. The chromatogram is developed using water-saturated butanol, whereby 100 ml fractions are separated and evaporated in a high vacuum at a bath temperature not exceeding 50°C. The middle fractions (3.35 g), which give a positive color reaction with Keller's reagent, are purified again by chromatography on cellulose powder. When the chromatogram is developed using water-saturated butanol, 50 ml fractions are separated and, after evaporation in a high vacuum at a bath temperature not exceeding 50°C, tested for the Keller color reaction. The following distribution is obtained: Fraction no. Residue Colour reaction mg Keller's 1 - 7 8 - 16 17 - 20 21 - 30 31 - 43 1270 87 79 1053 758 negative pure blue negative violet negative The residue from fractions 8-16 (87 mg) yields 45 mg of pure psilocin after treatment with silver carbonate as described in Example 2. Fractions 21-30 (1053 mg) yield 765 mg of pure psilocybin after treatment with silver carbonate. The active substances are obtained from the culture filtrate in the same manner. For this purpose the filtrate is concentrated in vacuo to about 1/10 of its volume. The accompanying substances are precipitated with ten times its volume of methanol, then filtered, and the solution is evaporated to dryness in a vacuum, and the residue is extracted three times with ten times its volume of methanol. The residue after evaporation of the methanol extracts is further processed as above. From 12 liters of culture filtrate, 80 mg of psilocybin and 6 mg of psilocin are obtained. Overall yield: 845 mg of psilocybin and 51 mg of psilocin, which corresponds to a content of 0.28% or 0.017% based on the dry mushroom material. Example IV. Obtaining psilocybin and psilocin from pure cultures of Psilocybe mexicana Heim. This example provides a detailed description of the in vitro cultivation of Psilocybe mexicana Heim mushrooms to produce mycelium and sclerotia, as well as the preparation of pure active substances. Fresh, hop-free beer wort is diluted with water to a dry matter content of 4.0-4.5%. For each liter of this solution add: FeSO4-7H2O 0.00417 g ZnSO4-7H2O 0.00172 g agar-agar 2.0 g This medium is poured in 500 ml portions into 1.6 liter Fernbach flasks and sterilized in an autoclave for 25 minutes at 108°C. After cooling, the contents of the flasks are inoculated with 2 ml of a suspension of the Psilocybe mexicana Heim mushroom. The vaccine is obtained by placing spores of pure cultures of the above-mentioned cap fungi on beer wort and agar. The spores falling from the gills of the mature pericarp are introduced onto a nutrient medium made of beer wort and agar and allowed to mature. The vaccine can be obtained from the primary cultures thus obtained in the following way: the mycelium is scraped with a sharp spatula under a sterile water supply so that a suspension of fine mycelial flocs is formed. This vaccine is inoculated into the nutrient medium placed in Erlenmeyer flasks (in which saddle-shaped porcelain filling shapes are placed). The experiments are best carried out with Erlenmeyer flasks containing 50 g of saddle-shaped filling shapes (size - 1 cm, weight about 0.9 g) and 80 ½ ml of medium, consisting of beer wort with a dry matter content of about 4% and 0.5% - 6% agar. During maturation at 24°C, after two weeks, a tight mycelium cover forms on the surface. The entire culture is centrifuged for 30 minutes on a shaker, with saddle shapes grinding the mycelium so that a fine suspension of mycelium flocs is formed. The inoculum thus obtained is sufficient to inoculate 25 liters of medium. The inoculated cultures mature in the dark at a temperature of 24-26°C. This creates the already described mycelium cover, obliterating the very numerous sclerotia, which generally reach a size of about 1 cm (in individual cases they may be significantly larger). (larger ones). To separate the mycelium and sclerotia, pure cultures are filtered through gauze, pressed and dried in an oven at 35-40°C. From one batch of 135 Fernbadh flasks containing 67 liters of medium, 1149 g of dry material (sclerotia and mycelium) is obtained after harvesting after 62 days, which corresponds to 17.14 g per liter of adjusted medium. To obtain the active substances, for example 612 g of dried material consisting of sclerotia and mycelium is finely powdered and extracted three times with 1 liter of chloroform and another three times with 1 liter of chloroform with the addition of 10% ethanol. 5.6 g is transferred to the extract. inactive accompanying substances. The pre-extracted fungal material is then thoroughly leached with methanol (once in 6 liters and three times in 3 liters). The combined methanol extracts are evaporated under reduced pressure to dryness, leaving 35 g of a light brown residue. This residue is poured with 35 ml of water to remove fatty impurities and shaken once with 1 liter and twice with 0.5 liters of petroleum ether. The petroleum ether now contains 1.5 g of inactive accompanying substances. The remaining aqueous solution is first concentrated in vacuo to about 50 ml and then, with vigorous stirring, 500 ml of anhydrous ethanol is added. The solution containing the active substances The precipitate is separated by decantation from the resulting sticky precipitate. The precipitate is dissolved again in a small amount of water and again treated with ten times the same amount of absolute alcohol. This precipitation operation is repeated two more times. The solutions are combined and evaporated in a vacuum to dryness. A solid residue of 23.4 g is obtained, which contains the total amount of active substances. For chromatography on a cellulose column, the precipitate is dissolved in the smallest possible amount of 50% methanol, and the solution is thoroughly mixed with 80 g of cellulose powder, after which the material thus obtained is dried in a vacuum. The cellulose powder, thus loaded with the substance, is introduced into a cellulose column, the packing of which was prepared by desliming 700 g of cellulose powder using The chromatogram is developed with water-saturated butanol. The chromatogram is developed with water-saturated butanol, 200 ml fractions being separated and evaporated in a high vacuum at a bath temperature not exceeding 50°C. The following distribution is obtained: Fraction No. 1-14 15-34 35-40 Table 1 Residue S 2.170 6.660 3.540 12.370 Keller's color reaction negative positive - negative The residue, approximately 11 g, is retained on the column. Fractions 15-34, which yield a total of 6.660 g of residue, contain a certain amount of active substance. For further purification, they are again chromatographed as described below. The 6.6 g of residue is dissolved in 20 g of cellulose powder is saturated with the smallest possible amount of methanol, the resulting solution is dried and then introduced into a column filled with 600 g of cellulose powder treated as above. When the chromatogram is developed with butanol saturated with water, 100 ml fractions are separated, which, after evaporation in a high vacuum, are tested for color reaction. The following distribution is obtained: Table 2 Fraction No. 1-8 9-17 18-21 22-31 32-45 Residue mg 2.950 180 155 912 1.510 Keller's color reaction negative pure blue negative violet negative """ Residue fraction 9-17 (180 mg) yields, after treatment with silver carbonate as described in Example 1, 90 mg of pure psilocin. Fractions 22-31 (Table 2) yield, after treatment with silver carbonate as described in Example 1, 619 mg of pure psilocybin with the properties described in Example 1. The active substances are obtained from the culture filtrate in the same manner as described above for the sclerotia. For this purpose, the culture filtrate is concentrated in a vacuum to about 1/10 of its volume, treated with ten times the volume of methanol, filtered, and the residue is extracted three times with ten times the volume of methanol. The residue after evaporation of the methanol extracts is further processed as described above. From 10 liters The culture filtrate yields, for example, 220 mg of psilocybin and 12 mg of psilocin. Example V. Obtaining psilocybin and psilocin from pure cultures of Stropharia cubensis Earle. The nutrient solution is prepared as follows: fresh, light beer wort without the addition of hops is diluted with running water to 6% of dry substance content. To each liter of this solution the following is added: FeS04-7H20 0.0021 g ZnS04.7H20 0.0009 g Ca/N03/2 1.0 g KH2P04 0.0624 g MgS04.7H20 0.0624 g KC1 0.0312 g Agar-agar 2.0 g This medium is sterilized as in Example III and inoculated with 2 ml of Stopharia cubensis Earle mushroom suspension per liter of medium. The preparation of this inoculum in suspension is carried out as described in Example III. After 52 days of maturation in the dark at a temperature of 24°C, 452 g of dry mushroom material are obtained from a 20 liter load, i.e. 22.6 g per liter of medium. The preparation of the active substances from the mushroom material is carried out as described in Example III. Yield: 1083 mg of pure crystalline psilocybin and 45 mg of psilocin, which corresponds to a yield of 0.24% or 0.010% based on the dry mushroom material. Example VI. Obtaining psilocybin and psilocin from Psilocybe semperviva Heim et Cailleux of natural origin. Ripe mushroom fruiting bodies Psilocybe semperviva Heim et Cailleux obtained by artificial cultivation in natural medium (compost) is carefully dried in a stream of air at 30°C. 32 g of dried pericarp is finely ground and shaken at room temperature for 1/2 hour, once with 300 ml of methanol and three times with 150 ml of methanol each. The combined extracts are evaporated in a vacuum to dryness. The residue (3.3 g) is degreased by trituration four times with 125 ml of petroleum ether and three times with 50 ml of chloroform containing 10% ethanol. 6.5 g of residue is obtained, which is dissolved in 6 ml of water. This solution is slowly treated with 60 ml of anhydrous ethanol to precipitate any remaining accompanying substances, thereby enriching the solution in active substances. This purification is repeated two more times. The solutions are decanted, combined, and evaporated in a vacuum to dryness. The residue is dissolved in methanol and chromatographed on cellulose powder as described in Example II. The active substances thus obtained are freed from halogen by treatment with silver carbonate. After recrystallization, 160 mg of crystalline psilocybin and 32 mg of psilocin are obtained, which corresponds to a content of 0.5% or 0.1% based on the dried mushroom material. Example VII. Obtaining psilocybin from Psilocybe caerulescens Murrill var. Mazatecorum Heim of natural origin. Ripe fruiting bodies of the mushroom Psilocybe caerulescens Murrill var. Mazatecorum Heim obtained By artificial cultivation on a natural medium (compost), it is carefully dried, then the dried mushroom material (7.3 g) is finely ground and thoroughly extracted with methanol, as described in Example II. The extracts are combined and evaporated in a vacuum to dryness. The residue obtained as in Example II is further processed to obtain the active substances. 14.6 mg of psilocybin is obtained (referring to the dried mushroom material). No psilocin is obtained from the processed mushroom sample. Example VIII. Obtaining psilocybin from pure cultures of Psilocybe carulescens Murrill var. Mazatecorum Heim.* The culture medium is prepared by diluting fresh beer wort without added hops with running water to a dry matter content of 4.0%. each liter of this solution is added: corn liquor 10.0 g FeS04-7H20 0.00834 g Ca/OH/2 0.20 g K2HP04 0.20 g NH4OH 0.25 g Agar-agar 2.0 g - 8 -The medium is sterilized as indicated in Example III, and inoculated with a mycelium suspension pure breeding Psilócibe caerulescens Murrill var. Heim mazate-cerum from Mekhyk, and the cultures produced are matured in the dark at a temperature of 26°C. After 48 days, 20.4 g of dry mushroom material per liter of solution have accumulated. The active substances are isolated and purified from this material as described in Example III. Efficiency per load; With 10 liters of medium contains: 449 mg of psilocybin, which corresponds to a content of 0.23% of the dried mushroom material. No psilocin was found in this load. Example IX. Obtaining psilocybin from Psilccibe Zapotecorum Heim of natural origin. Ripe fruiting bodies of Psilccibe Zapotecorum Heim collected and sorted in Mexico in the area known as "pays chatino" are carefully dried (residue 42.4 g), finely ground and shaken with methanol as described in Example II. The combined methanol extracts are evaporated in a vacuum to dryness. The residue is further processed as described in Example II. 212 mg of pure psilocybin are obtained, which corresponds to a content of 0.5%. No psilocybin is obtained from the processed mushroom material. Psilocin. Example X. Obtaining psilocybin from pure cultures of Psilócibe Zapotecorum Heim. Fresh, light beer wort without added hops is diluted with running water to 4.5% dry matter. To each liter of this solution add: * FeSO4 corn steep liquor * NH4OH * K2HPO4 * Agar-agar * pH of the solution: 5.5. This culture medium is sterilized as described in Example III and inoculated with a mycelial suspension of a pure culture obtained from the spores of a mature pericarp of Psilócibe Zapotecorum Heim from "pays chatamo" in Mexico. After 57 days of dark maturation at 24°C, 430 g of dried mushroom material are obtained from a 25 liter load, i.e., 17.2 g per liter of medium. The active substances were obtained from this material according to the method given in Example III. Yield: 903 mg of pure psilocybin, which corresponds to a content of 0.21% based on the dry mushroom material. Psilocybin is not obtained. Example XI. Obtaining psilocybin and psilocin from Psilócibe aztecorum Heim of natural origin. Ripe fruiting bodies of Psilócibe aztecorum Heim collected in Mexico in the country of the Aztecs (Na Popocatepetl, at an altitude of 3300-3500 m above sea level) are carefully dried, finely ground, and thoroughly extracted with methanol as described in Example II. The extracts are combined and evaporated to dryness in a vacuum. The residue is treated to isolate the active substances as given in Example II. 570 mg of pure psiiocybin and 47 mg of psilocin are obtained, which corresponds to a content of 0.2% or 0.017% based on dry mushroom material (285 g). Example XII. Obtaining psiiocybin and psilocin from pure cultures of Psilócibe Aztecorum Heim. The medium includes: 0.00834 g 10.0 0.30 0.30 2.0 g g era malt extract - FeS047H20 ZnOS4-7H,C Ca/NOjj/a KH2P04 MgS04-7H20 KC1 Agar-agar 100 g 0.00417 g 0.00172 g 1.0 g 0.25 g 0.25 g J 0.125 g 2.0 g running water to 1000 ml The medium is sterilized in an autoclave for 25 minutes at 108°C. 1 liter is inoculated with 2 ml of a suspension of the fungus Psilócybe aztecorum Heim. The preparation of this suspension inoculum is carried out as described in Example 31a. The cultures are matured in the dark for 45 days at 24°C and harvested as described in Example 31c. *The yield from 10 liters of medium is 86.5 g of dry mycelium. The finely ground mycelium is shaken at room temperature for 1/2 hour with 1 liter of 80% ethanol. The residue after filtration is extracted three more times in the same way. The residue obtained by evaporation of the combined The extracts (8.9 g) were leached successively, in order to separate them from fatty accompanying substances and inactive ballast substances, twice with 100 ml of petroleum ether, twice with 80 ml of chloroform, and twice with 50 ml of acetone. 5.6 g of water-soluble powder were obtained, which was dissolved in 6 ml of water. The solution was slowly stirred and treated with 80 ml of acetone, the precipitate separated, which was then dissolved in a small amount of water and precipitated again with a tenfold amount of acetone. This purification by precipitation was repeated two more times, with some of the tin being transferred to the hydrogen-acetone extract. The extracts were evaporated in vacuo, and the residue (2.8 g) was purified and separated by column chromatography. with cellulose powder. After chromatography twice, 225 mg of crystalline psilocybin and 15 mg of psilocin are obtained, which corresponds to a yield of 0.26% or 0.017% based on dry mushroom material. Example XIII. Preparation of culture. The medium is obtained as follows: Fresh beer wort (light without hops) is diluted with running water to a dry substance content of 4.0 to 4.5%. To one liter of this solution is added: FeS04.7H20 0.00209 g ZnS04-7H20 0.00086 g Ca/N03/2 0.25 g KH2P04 0.0625 g MgS04 0.0625 g KC1 0.031 g Agar-agar 2.0 g This medium is sterilized and inoculated as in Example 4. After 48 days of maturation at 24°C, a yield of 924 g of sclerotia and mycelium (dry matter) is achieved from one load of 55 liters, which is 16.6 g per liter of medium. The active substances are obtained from this material as described in Example 4. Yield: 0.018% psilocin and 0.12% psilocybin, based on the dry mushroom material. Example 14. Obtaining a culture. The medium is prepared as follows: Fresh beer wort (light, without hops) is diluted with running water to a dry matter content of 4.0-4.5%. For each liter of this solution, add: FeSO4.7H2O 0.00209 g ZnSO4.7H2O 0.00086 g Ca/NOa/a 1.0 g Agar-agar 2.0 g The medium is sterilized, inoculated and matured as in Example 4. After 48 days, 20.5 g of dry mycelium and sclerotia are collected per liter of medium, from which the active substance is isolated as in Example 4. Yield: 0.011% psilocin and 0.14% psilocybin in relation to the dried mushroom material. Example 15. Obtaining a culture. The medium is prepared as follows: Fresh beer wort (light, without hops) is diluted with running water to a dry matter content of 4.0-4.5%. To each liter of this solution is added: FeSO4.7H2O 0.00209 g ZnSO4.7H2O 0.00086 g KH2PO4 0.25 g Agar-agar 2.0 g Further processing is carried out as described in Example 4. The yield of dried sclerotia and mycelium is 15.9 g per liter of medium after 48 days. The isolation of the active substances is carried out as described in Example 4. Yield: 0.02% psilocin and 0.11% psilocybin in relation to the dried mushroom material. Example XVI. Obtaining a culture. The medium is prepared as follows: Fresh beer wort (light, without hops) is diluted with running water to a dry matter content of 4.0-4.5%. To each liter of this solution add: FeS04.7H20 0.00209 g ZnS04.7H20 0.00086 g corn soak 20.0 g Agar-agar 2.0 g Further processing is carried out as described in Example IV. The yield of dried sclerotia and mycelium after 48 days is 29.3 g per liter of medium. The separation of active substances is carried out as described in Example IV. Yield: 0.012% psidocin and 0.22% psilocybin in relation to dry mushroom material. Example XVII. Obtaining culture. The medium used is: malt extract 45 g FeS04-7H20 0.00417 g ZnS04.7H20 0.00172 g running water 1000 ml of the medium is sterilized in an autoclave at 108°C for 25 minutes. 1 liter of the medium is inoculated with 10 ml of a suspension of the Psilocybe mexicana mushroom, the preparation of the suspension being carried out as described in Example IV. After inoculation, the fungus is propagated in this medium at 24°C. This produces sago-like mycelium balls, as is known in the propagation of lower fungi. After 30 days, the mycelium is separated by filtration. 7.0 g of dry mycelium are obtained per liter of medium. 430 g of dried, finely ground mycelium are shaken at room temperature for 1/9 hour with 4.5 liters of 80% ethanol. After filtration, the residue is extracted in the same way. The same procedure was repeated three more times. The residue after evaporation of the combined extracts (41 g) was leached successively twice with 500 ml of petroleum ether, twice with 400 ml of chloroform, and twice with 200 ml of acetone to separate the ballast substances. 25 g of water-soluble powder remained, which was dissolved in 25 ml of water. 250 ml of acetone was slowly added to the solution with good stirring, and the precipitate, after separating from the liquid, was dissolved again in a small amount of water and reprecipitated with a tenfold amount of acetone. These operations were repeated twice more with the acetone-insoluble precipitate, with all the active substances transferred to the water-acetone extracts. These extracts were evaporated with good stirring, and the residue (9.8 g) was further separated. by chromatography on a cellulose column as described in Example IV. After chromatography twice, 32 mg of crystalline psilocin and 225 mg of psilocybin are obtained, which corresponds to a yield of 0.007 or 0.05% based on dry mycelium. PL PL