KR100991375B1 - Novel clitocybin derivates, preparation method thereof and composition containing the same for prevention of aging as an active ingredient - Google Patents

Abstract

The present invention relates to a novel crytocycline derivative represented by the following Chemical Formula 1, a method for preparing the same, and an anti-aging composition containing the same as an active ingredient. The novel crytocybin derivative according to the present invention exhibits excellent radical scavenging activity. In addition, it shows the effect of reducing DNA damage and reducing erythema by ultraviolet rays, so it can be useful for preventing and treating diseases caused by excessive accumulation of active oxygen as well as for preventing aging. It can be usefully used as a composition or a functional cosmetic composition.

[Formula 1]

(In Formula 1, R ¹ and R ² are as defined in the specification.)

 Oriole mushroom, antioxidant, anti-aging, DNA damage suppression, erythema reduction, sun protection, functional cosmetics

Description

Novel clitocybin derivates, preparation method approximately and composition containing the same for prevention of aging as an active ingredient

The present invention relates to a novel isoindoleone-based crytocycline derivative, a preparation method thereof and an anti-aging cosmetic composition containing the same as an active ingredient.

By a variety of oxidation reactions, chemicals, food, human diseases and radiation superoxide (superoxide, O ₂ ^-), hydroxyl radicals (hydroxyl radical, · OH), and a large strong reactivity such as hydrogen peroxide (H ₂ O ₂₎ When active oxygen and free radicals are generated in vivo, lipids of cell membranes containing a large amount of unsaturated fatty acids are oxidized, thereby producing lipid peroxides on the cell membranes. Accumulation of lipid peroxides in the cell membrane reduces the fluidity and functionality of the cell membrane, thereby inhibiting the overall function of the cell and changing the structure of the cells. In addition, free radicals and free radicals modify or destroy cell constituents such as nucleic acids and sugars, resulting in cancer, liver disease such as alcoholic hepatitis, stroke due to cerebrovascular disorders, myocardial infarction, diabetic vascular disorders, hyperlipidemia, and acute inflammation. , Rheumatic diseases and other human diseases . [ Arthritis. Res. Ther. 6 (6): 265-78 (2004), J. Neuropathol. Exp. Neurol. 65 (7): 631-41 (2006), Trends. Mol. Med. 12 (11): 521-8 (2006), Int. J. Biochem. Cell. Biol. 39 (1); 44-84 (2007). In addition, reactive oxygen reacts with unsaturated fatty acids, which are present in a large amount in foods, which causes rancidity, which may result in deficient defects in food safety and quality maintenance, and oxidative stress of plants and animals caused by various oxides, and active oxygen generated during fermentation of microorganisms. In many fields, such as yield reduction caused by free radicals are causing problems.

Therefore, the discovery of a new substance with powerful yet non-toxic antioxidant activity will be very useful as the development of a therapeutic agent for various diseases or as an anti-aging cosmetic additive and food additive.

Until now, synthetic antioxidants such as butylated hydroxy toluene (BHT) and butylated hydroxy anisol (BHA) have been used as lipid peroxidation inhibitors. However, these BHT and BHA have excellent lipid peroxidation inhibitory activity, but are likely to cause cancer or malformation. There was a disadvantage that can not be used very continuously. Therefore, it is necessary to create an antioxidant active substance from natural products and develop an antioxidant having no toxicity or side effects.

On the other hand, about 20 species of antioxidants detected from microbial metabolites are known, and most of the active antioxidants derived from microorganisms reported so far are derived from Streptomyces , and carbazole-based The most reported compound. Representative compounds include Streptomyces In addition to carquinostatin found in acetone extract of exfoliatus cells, antiostatin, carazostatin, neocarozostatin, and carbazomycin have been reported. In addition, benthocyanin, benthophoenin, and phenazoviridin have been identified as phenazine-based antioxidants. Other thiazole-based compounds include thiazolstatin and naphterpine blood. Rolostatin and the like have been reported.

Most of the researches on the detection of lipid peroxidation inhibitors in Korea are limited to the use of food materials to measure the presence and activity of antioxidant activity and use as a food protection agent. Therefore, the researches on the extraction, purification and structural analysis of active substances are very insufficient. It was.

In the early 1990s, the Korea Research Institute of Bioscience and Biotechnology began to search for lipid peroxidation inhibitors in earnest, investigating lipid peroxidation inhibitors from soil actinomycetes or natural products. Novel indole alkaloid compounds (J. of Natural Products, 60: 721-723, 1997), showing active oxygen removal activity from Agrocybe cylindracea mushrooms, new antioxidant compounds derived from soybean mushrooms, and containing them as active ingredients Antioxidant composition (Korean Patent Registration No. 0466031, 2005), a new hispidine-based active oxygen scavenging substance and its manufacturing method (Korean Patent Registration No. 0320787, 2002), Terphenyl-based novel from Polyozellus multiplex strain Compound Polygelline (Korea Patent Registration 0362394, 2002), Wrinkle Teacup Mushroom Mycelia Antioxidant activity extract, and a method of producing from the culture broth (Republic of Korea Patent Application No. 2006-96391 Ho, 2006) has developed a bar or the like.

The present inventors continue to search for new and powerful antioxidant active substances from Korean native mushrooms, and iso-indoleone-based compounds exhibiting new and potent antioxidant activity not yet identified from the mycelium culture extract of Hygrophoropsis aurantiaca KCTC 11143BP. Was found and named Crytocybin A, and after synthesizing Crytocybin B and C in organic synthesis, it was confirmed that they exhibit potent antioxidant activity and completed the present invention.

It is an object of the present invention to provide novel crytocybin derivatives.

It is another object of the present invention to provide a method for preparing the crytocybin derivative.

Still another object of the present invention is to provide an antioxidant containing the novel chrythocybin derivative as an active ingredient.

It is another object of the present invention to provide a sunscreen agent containing the novel crytocycline derivative as an active ingredient.

Still another object of the present invention is to provide a pharmaceutical composition for preventing and treating diseases caused by excessive accumulation of active oxygen, which contains the novel crytocybin derivative as an active ingredient.

In addition, the present invention is to provide an anti-aging pharmaceutical composition containing the novel chrythocybin derivative as an active ingredient.

Still another object of the present invention is to provide an anti-aging cosmetic composition containing a novel crytocybin derivative as an active ingredient.

In order to achieve the above object, the present invention provides a novel crytocybin derivative.

The present invention also provides a method for producing the novel crytocybin derivative.

Furthermore, the present invention provides an antioxidant containing the novel chrythocybin derivative as an active ingredient.

In addition, the present invention provides a sunscreen containing the novel chrythocybin derivative as an active ingredient.

Furthermore, the present invention provides a pharmaceutical composition for the prevention and treatment of diseases caused by the accumulation of excess active oxygen containing the novel cryocybin derivative as an active ingredient.

In addition, the present invention provides an anti-aging pharmaceutical composition containing the novel chrythocybin derivative as an active ingredient.

In addition, the present invention provides an anti-aging cosmetic composition containing a novel chrythocybin derivative as an active ingredient.

The novel cryocybin derivatives according to the present invention exhibit excellent radical scavenging activity, and in addition to the effect of reducing DNA damage, reducing erythema by ultraviolet light, preventing and treating diseases caused by excessive accumulation of free radicals. In addition, it may be usefully used for anti-aging, and may be usefully used as an anti-aging pharmaceutical composition or functional cosmetic composition using the same.

The present invention provides a novel crytocybin derivative represented by the following formula (1).

(In Formula 1, R ¹ and R ² are independently or optionally OCH ₃ or OH.)

To more specifically exemplify the novel crytocycline derivative represented by the formula (1) is the same as the formula (1a-1c).

The compound of Chemical Formula 1a according to the present invention exhibits the appearance of light brown powder, and the molecular formula is represented by C ₁₄ H ₁₁ O ₄ N, and was named cryocybin A compound.

In addition, the compound of Formula 1b according to the present invention exhibits the appearance of a pale yellow powder, and the molecular formula is represented by C ₁₅ H ₁₃ O ₄ N, and was named cryocybin B compound.

Furthermore, the compound of Formula 1c according to the present invention exhibits the appearance of a white powder, the molecular formula is represented by C ₁₆ H ₁₅ O ₄ N, it was named Kratocyvin C compound.

The compound of Chemical Formula 1 according to the present invention may be preferably obtained by extracting and purifying from an Oriole mushroom strain, and may be obtained through organic synthesis.

The compound of Chemical Formula 1 of the present invention may be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include those derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid, and aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxyalkanoates, Dioleate, aromatic acid, aliphatic and aromatic sulfonic acids. Such pharmaceutically nontoxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, and iodide. Id, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suverate , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, meth Oxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesul Nate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1- Sulfonates, naphthalene-2-sulfonates or mandelate.

The acid addition salts according to the invention can be dissolved in conventional methods, for example, by dissolving the compound of Formula 1 in an excess of aqueous acid solution, which is then subjected to a water miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. Can be prepared by precipitation.

The same amount of the compound of formula 1 and the acid or alcohol in water may be heated and then the mixture is evaporated to dryness or prepared by suction filtration of the precipitated salt.

In addition, bases can be used to make pharmaceutically acceptable metal salts. Alkali metal or alkaline earth metal salts are obtained, for example, by dissolving a compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically suitable to prepare sodium, potassium or calcium salt as the metal salt. Corresponding silver salts are also obtained by reacting alkali or alkaline earth metal salts with a suitable negative salt (eg, silver nitrate).

The present invention includes not only the compound of Formula 1 or a pharmaceutically acceptable salt thereof, but also all possible solvates and hydrates that can be prepared therefrom.

In addition, the present invention provides a method for preparing the novel crytocybin derivative.

The novel chrytocybin derivatives according to the present invention can be prepared by extraction and purification from oriole mushrooms and by organic synthesis.

In one embodiment of the method for producing a novel crytocybin derivative according to the present invention

Obtaining a culture solution by culturing the hygrophoropsis aurantiaca KCTC 11143BP strain (Step 1);

Extracting the culture solution obtained in step 1 with acetone and ethyl acetate (step 2);

Obtaining an active fraction by performing concentration gradient silica gel column chromatography on the extract extracted in step 2 using a mixed solvent of chloroform and methanol (step 3); And

The active fraction obtained in step 3 is subjected to Sephadex LH-20 column chromatography using methanol (100%) as a solvent to obtain a crytocybin derivative of Chemical Formula 1 (step 4).

First, step 1 is a step of obtaining a culture by culturing the strains of Hygrophoropsis aurantiaca KCTC 11143BP.

At this time, the culture solution is preferably obtained by incubating the strain of Hygrophoropsis aurantiaca KCTC 11143BP in a yeast-malt extract (Yeast extract, YPS) liquid medium.

Next, step 2 is a step of extracting the culture solution obtained in step 1 with acetone and ethyl acetate.

The culture solution obtained in step 1 is preferably extracted with acetone and then re-extracted with ethyl acetate. Extraction method may be used an extraction method commonly used in the art, the ethyl acetate layer is used in the next step after re-extraction.

Next, step 3 is a step of performing a concentration gradient silica gel column chromatography using a mixed solvent of chloroform and methanol the extract extracted in step 2.

At this time, the concentration gradient of the mixed solvent of chloroform and methanol is preferably 50: 1 ~ 1: 1, eluting at a rate of 10 ~ 15 ㎖ / min to obtain an active fraction after 150 minutes.

Next, step 4 is a step in which the active fraction obtained in step 3 is subjected to Sephadex LH-20 column chromatography using methanol (100%) as a solvent to obtain a cryotocycline derivative of Chemical Formula 1.

The active fraction obtained in step 3 was eluted at a rate of 1-1.6 ml / min using methanol (100%) as eluent using Sephadex LH-20 column chromatography. An empty derivative (Formula 1a) can be obtained.

As another embodiment of the method for preparing a novel crytocybin derivative according to the present invention, as shown in Scheme 1 below,

Reacting the compound of formula 3 with the compound of formula 2 in an organic solvent to prepare a compound of formula 1 (Formula 1c) (step a).

(In Formula 1, Formula 1c is included in Formula 1 of the present invention.)

In addition, as another embodiment of the method for producing a novel crytocybin derivative according to the present invention, as shown in Scheme 2 below,

Preparing a compound of formula 1c by reacting the compound of formula 3 with a compound of formula 2 in an organic solvent (step a); And

Bromic acid and acetic acid are added to the compound of Formula 4 prepared in step a, followed by extraction with ethyl acetate to prepare the compound of Formula 1 (Formula 1a and Formula 1b) (step b).

(In Formula 2, Formula 1a and Formula 1b are included in Formula 1 of the present invention.)

First, step a is a step of preparing a compound of Formula 1c by reacting a compound of Formula 3 with a compound of Formula 2 under an organic solvent.

In this case, the organic solvent used is preferably methanol, and the mixing ratio of the compound of Formula 2 and the compound of Formula 3 is preferably 1: 1 equivalent.

Next, step b is a step of preparing the compound of Formula 1 (Formula 1a and Formula 1b) by adding bromic acid and acetic acid to the compound of Formula 1c prepared in step a and extracted with ethyl acetate.

Specifically, the aqueous bromic acid solution and acetic acid are added to the compound of Formula 1c prepared in step a and refluxed for 15 to 20 hours, and then the reaction solution is poured into ice water and extracted with ethyl acetate. The organic layer was washed with water, dried over anhydrous magnesium sulfate and concentrated under reduced pressure, and then purified by silica gel column chromatography (dichloromethane: methanol = 10: 1) to obtain at least about 80% of the crytocybin derivatives of the formulas (1a) and (1b). Can be.

Furthermore, the present invention provides an antioxidant containing the novel chrythocybin derivative as an active ingredient.

Radical scavenging activity of superoxide, DPPH and ABTS ⁺ was investigated using the novel crytocybin derivatives according to the present invention.The radical scavenging activity was superior to that of the control trolox, BHA, ferulic acid or catechin. It was confirmed that (see Table 3).

In addition, the effect of reducing intracellular DNA damage was confirmed that the number of tail DNA generated during hydrogen peroxide treatment was significantly reduced by administering the novel crytocybin derivative (see FIG. 8 and Table 4).

Through this, the novel chrythocybin derivative of the present invention can be usefully used as an antioxidant.

In addition, the present invention provides a sunscreen containing the novel chrythocybin derivative as an active ingredient.

In the UV-blocking effect experiment using the novel crytocybin derivatives according to the present invention, the novel crytocybin derivatives according to the present invention resulted in the reduction of erythema similarly to the control group known to have excellent erythema-reducing effect against ultraviolet irradiation. (See FIG. 9 ). Through this, the novel crytocycline derivative of the present invention can be usefully used as a sunscreen.

Furthermore, the present invention provides a pharmaceutical composition for the prevention and treatment of diseases caused by the accumulation of excess active oxygen containing the novel cryocybin derivative as an active ingredient.

In addition, the present invention provides an anti-aging pharmaceutical composition containing the novel chrythocybin derivative as an active ingredient.

Isolated from melodious great mushrooms (Hygrophoropsis aurantiaca KCTC 11143BP) of the present invention, a purified novel Cry Sat Im empty derivatives superoxide (superoxide), DPPH and ABTS ⁺ the trawl control from a radical scavenging activity was measured as the target rocks, BHA, It was confirmed that it shows superior radical scavenging activity than ferulic acid or catechin (see Table 3). In addition, the effect of reducing intracellular DNA damage was confirmed that the number of tail DNA generated during hydrogen peroxide treatment was significantly reduced by administering the novel crytocybin derivative (see FIG. 8 and Table 4). Highly reactive free radicals such as hydrogen peroxide and free radicals damage DNA and the like, and the damage of the DNA promotes aging [ Free Radic. Biol. Med. 43: 477-503 (2007). Therefore, the excellent radical scavenging activity and reduced DNA damage indirectly indicate the production of free radicals.

Therefore, the pharmaceutical composition of the present invention inhibits the production of free radicals, hepatic diseases such as alcoholic hepatitis caused by excessive accumulation of free radicals, stroke due to cerebrovascular disorders, myocardial infarction, diabetic vascular disorders, hyperlipidemia, acute Inflammation, rheumatism and cancer etc. Arthritis . Res . Ther . 6 (6): 265-78 (2004), J. Neuropathol . Exp . Neurol. 65 (7): 631-41 (2006), Trends . Mol . Med . 12 (11): 521-8 (2006), Int . J. Biochem . Cell . Biol . 39 (1); 44-84 (2007)] may be useful for the prevention and treatment of various human diseases as well as for the prevention of aging.

The composition of the present invention may further contain one or more active ingredients exhibiting the same or similar functions in addition to the novel chrythocybin derivatives.

The composition of the present invention may be administered in various oral and parenteral dosage forms during actual clinical administration, and when formulated, diluents or excipients such as fillers, weights, binders, wetting agents, disintegrating agents, surfactants, etc., which are commonly used, may be used. Are prepared using. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, which may contain at least one excipient such as starch, calcium carbonate, gelatin, etc. Mix and quiet. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, and lyophilized preparations. As the non-aqueous solvent and suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used.

Dosage ranges according to the patient's worm, age, sex, health condition, diet, time of administration, method of administration, rate of excretion and the severity of the disease. The daily dosage of the crytocycline derivative according to the present invention is about 0.1 to 100 mg / kg, preferably 5 to 50 mg / kg, and more preferably administered once to several times a day.

As a result of toxicity experiments by orally administering the novel crytocybin derivatives of the present invention to rats, 50% lethal dose (LD ₅₀ ) of the crytocybin derivatives is at least 1,000 mg / kg, which is considered a safe substance.

The composition of the present invention can be used alone or in combination with methods of using surgery, hormonal therapy, drug treatment and biological response modifiers to prevent aging as well as the prevention and treatment of diseases caused by excessive accumulation of free radicals. have.

In addition, the present invention provides an anti-aging cosmetic composition containing the novel cryocybin derivative as an active ingredient.

Anti-aging cosmetics of the present invention include, but are not limited to, lotions, ointments, gels, creams, patches or sprays. In preparing the anti-aging cosmetics containing the novel cryocybin derivatives of the present invention, 1 to 15 parts by weight, preferably 2 or 3 parts of the novel crytocycline derivatives of the present invention are included in the external preparation for skin. 10 parts by weight may be added. The skin external preparations of the present invention include fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, in addition to the novel crytocybin derivatives of the present invention. For surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or skin It may contain adjuvants commonly used in the field of dermatology, such as any other ingredients commonly used in external preparations. The ingredients may also be introduced in amounts generally used in the field of dermatology.

Hereinafter, the present invention will be described in detail by Examples, Experimental Examples and Formulation Examples.

However, the following Examples, Experimental Examples, and Formulation Examples specifically illustrate the present invention, and the content of the present invention is not limited to Examples, Experimental Examples, and Formulation Examples.

Example 1 Preparation of Crytocybin Derivatives 1 (Extracted, Isolated and Purified from Oyster Mushrooms)

Hygrophoropsis aurantiaca KCTC 11143BP strains were inoculated into a 500 ml Erlenmeyer flask containing 100 ml of yeast extract malt (YPS extract) in a reciprocating shaker at 27 degrees Celsius and 140 rpm. Incubated for Thereafter, 3 liter of yeast extract (Yeast extract, YPS) liquid medium was put into a 5 liter jar fermentor (Korea Fermentor Co. Ltd.) and sterilized, followed by inoculation of 100 ml of the shake cultured culture medium. Incubated under the same conditions. After culturing for 7 days, extracted with 70% acetone, re-extracted with ethyl acetate and separated into ethyl acetate layer and water layer.

The ethyl acetate extract obtained above was subjected to concentration gradient silica gel column chromatography using a mixed solvent of chloroform and methanol (50: 1 to 1: 1), followed by Sephadex LH-20 column chromatography (100% methanol). Was separated to give a light brown pure compound (6.5 mg). This was analyzed by reverse phase (ODS) column chromatography under 35% methanol solvent condition to confirm that the compound was 100% pure extracted and purified. The compound thus obtained was designated as Critocybin A.

< Example  2> Crytocyvin  Preparation of Derivatives 2 (Organic Synthesis)

(One) Crytocyvin  Preparation of C Compound

To a solution of methyl 2-peryl-3,5-dimethoxybenzoate (2,24 g, 10.0 mmol) in methanol (40 mL) was added 4-aminophenol (1.1 g, 10.0 mmol) for 1 hour. Stirred. Then, NaBH 4 (0.75 g, 20.0 mmol) was added to the reaction solution at 0 ° C., and the temperature was raised to room temperature, followed by stirring for 18 hours. The reaction solution was concentrated under reduced pressure, and water (50 mL) and a small amount of acetic acid were added thereto. The precipitate formed was filtered with a glass filter, and then the obtained solid was washed with water and dried to obtain the target compound CrytoSybin C compound (Formula 1c; 2.6 g, 91%).

¹ H NMR (300 MHz, DMSO-d6) δ 9.38 (s, 1H), 7.65 (s, 1H), 7.62 (s, 1H), 6.82-6.76 (m, 4H), 4.73 (s, 2H), 3.87 (s, 3H);

¹³ C NMR (75 MHz, DMSO-d6) δ 165.9, 161.4, 154.9, 154.2, 134.7, 131.1, 121.5, 115.2, 102.5, 97.8, 55.7, 55.6, 48.3

(2) Crytocyvin  Preparation of A Compound

To a crytocybin C compound (2.0 g, 7.0 mmol) prepared in (1) was added 48% aqueous bromic acid solution (10 mL) and acetic acid (20 mL) and refluxed for 18 hours. The reaction solution was poured into iced water and extracted with ethyl acetate. The organic layer was washed with water, dried over anhydrous magnesium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (dichloromethane: methanol = 10: 1) to obtain the target compound crytocybin A compound (1.5 g, 85%). It was.

¹ H NMR (300 MHz, DMSO-d6) δ 9.98 (s, 1H), 9.61 (s, 1H), 9.36 (s, 1H), 7.61 (dd, J = 6.6, 1.8 Hz, 2H), 6.79 (dd , J = 6.6, 1.8 Hz, 2H), 6.56 (dd, J = 1.8 Hz, 1H), 6.49 (dd, J = 1.8 Hz, 1H), 4.65 (s, 2H);

¹³ C NMR (75 MHz, DMSO-d6) δ 166.4, 161.4, 158.9, 154.1, 153.0, 134.9, 131.3, 121.5, 117.8, 115.2, 105.9, 100.1, 48.5

(3) Crytocyvin  Preparation of B Compound

To a crytocybin C compound (2.0 g, 7.0 mmol) prepared in (1) was added 48% aqueous bromic acid solution (10 mL) and acetic acid (20 mL) and refluxed for 18 hours. The reaction solution was poured into iced water and extracted with ethyl acetate. The organic layer was washed with water, dried over anhydrous magnesium sulfate, concentrated under reduced pressure, purified by silica gel column chromatography (dichloromethane: methanol = 10: 1), and together with the cryotocycline A compound, the target compound crytocybin B compound ( 0.76 g, 40%) was obtained.

¹ H NMR (300MHz, DMSO-d6) δ 9.85 (s, 1H), 9.36 (s, 1H), 7.62 (d, J = 9.3 Hz, 2H), 6.78 (d, J = 9.3 Hz, 2H), 6.67 (d, J = 1.8 Hz, 1H), 6.63 (d, J = 1.8 Hz, 1H), 4.70 (s, 2H), 3.84 (s, 3H);

¹³ C NMR (75 MHz, DMSO-d6) δ 166.0, 159.3, 154.9, 154.1, 134.7, 131.2, 121.5, 119.0, 115.2, 102.5, 100.5, 55.4, 48.3.

<Structure Analysis>

Physical and chemical properties of the compounds obtained in Examples 1 and 2 were measured as follows.

The fluorescence was measured using a polarizer (JASCO DIP-370 polarimeter). Ultraviolet absorption spectra were measured using an ultraviolet spectrophotometer (Shimadzu UV-260 spectrophotometer, Japan), and infrared absorption spectra were measured using an ultraviolet spectrophotometer (FT-IR Equinox 55 spectrometer, Burker, USA). In addition, HR-ESI-MS spectra were measured using a mass spectrometer (JEOL JMS HX-110 mass spectrometer, JEOL, Japan) to determine the molecular structure.

The measurement results are shown in Table 1.

Crytocycline A Compound Crytocycline B Compound Crytocycline B Compound Chemical structure

Appearance Light brown powder Light yellow White Optical intensity ^{[α] D 20 - 1.40 o} (c = 0.3) ^{[α] D 20 - 1.40 o} (c = 0.9) ^{[α] D 20 - 1.40 o} (c = 1.0) Chemical formula C ₁₄ H ₁₁ O ₄ N C ₁₅ H ₁₃ O ₄ N C ₁₆ H ₁₅ O ₄ N Molecular Weight 257 271 285 HR-ESI-MS m / z Found: 258.07571 (M + H ⁺ )
Calculation: 258.07608 Found: 272.0923 (M + H ⁺ )
Calculation: 272.0923 Found: 286.1079 (M + H ⁺ )
Calculation: 286.1079 UV absorption spectrum [UV λ _max nm (in methanol)] 211 (1.6), 286 (0.4) cm ^-1 211 (1.8), 286 (0.8) cm ^-1 211 (1.8), 286 (0.8) cm ^-1 Infrared absorption spectrum [R (cm ^-1 )] 3399, 2925, 1616, 1516, 1453, 1256, 1144, 1097 3399, 2925, 1616, 1516, 1453, 1256, 1144, 1097 3399, 2925, 1616, 1516, 1453, 1256, 1144, 1097 Availability MeOH, DMSO EtOAc, DMSO EtOAc, DMSO Insoluble Hexane, water Hexane, water Hexane, water

As shown in Table 1, the cryocybin A compound according to the present invention was obtained as a light brown powder, and the value of the high resolution ESI-MS analysis (M + H ⁺ ) was 258.07571 m / z, calculated as 258.07608 m / z. Almost similar to z, the molecular formula was determined to be C ₁₄ H ₁₁ O ₄ N. The UV spectra measured the maximum absorption wavelengths at 211 nm and 286 nm, and the IR spectra showed a band due to hydroxyl groups at 3399 cm ^-1 . Thus, the cryocybin A compound was identified as a novel iso-indoleone compound.

The cryocybin B compound according to the present invention was obtained as a pale yellow powder, and the high resolution ESI-MS analysis result (M + H ⁺ ) was 272.0923 m / z, which is consistent with the calculated 272.0923 m / z. The molecular formula was determined by C ₁₅ H ₁₃ O ₄ N. The UV spectra measured the maximum absorption wavelengths at 211 nm and 286 nm, and the IR spectra showed a band due to hydroxyl groups at 3399 cm ^-1 . Thus, the cryocybin B compound was identified as a novel iso-indoleone compound.

Crytocybin C compound according to the present invention was obtained as a pale yellow powder, and the results of high resolution ESI-MS analysis (M + H ⁺ ) were 286.1079 m / z, which is consistent with the calculated value of 286.1079 m / z. The molecular formula was determined by C ₁₆ H ₁₅ O ₄ N. The UV spectra measured the maximum absorption wavelengths at 211 nm and 286 nm, and the IR spectra showed a band due to hydroxyl groups at 3399 cm ^-1 . Thus, the cryocybin C compound was identified as a novel iso-indoleone compound.

In addition, ¹ H and ¹³ C NMR, HMQC ((1H-detected heteronuclear Multiplie-Quantum Coherence) of the z-Crytocybin A ~ C compound through nuclear magnetic resonance (NMR) analysis (Burker AMX 300, Burker, USA) Heteronuclear Multiple-Bond Cohence (HMBC) spectra were obtained, which are shown in FIGS.

Carbon number
(position) Crytocycline A Compound Crytocycline B Compound Crytocycline C Compound δC (ppm) δH (ppm) δC (ppm) δH (ppm) δC (ppm) δH (ppm) One 155 154.1 154.2 2 108 6.50 (d, J = 1.8 Hz) 102.5 6.63 (d, J = 1.8 Hz) 102.5 6.63 (d, J = 1.8 Hz) 3 162 159.3 161.4 4 102 6.71 (d, J = 1.8 Hz) 102.5 6.67 (d, J = 1.8 Hz) 97.8 6.99 (d, J = 1.8 Hz) 5 120 119.0 121.1 6 136 134.7 134.7 7 172 166.0 165.9 8 50 4.71 (s) 48.3 4.70 (s) 48.3 4.68 (s) 9 133 131.2 131.1 10,14 124 7.54 (d, J = 9.0 Hz) 121.5 7.62 (d, J = 9.3 Hz) 121.5 7.68 (d, J = 9.0 Hz) 11,13 117 6.84 (d, J = 9.0 Hz) 119.0 6.78 (d, J = 9.3 Hz) 115.2 6.89 (d, J = 9.0 Hz) 12 157 154.9 154.9 15 55.4 3.84 (s) 55.6 3.83 (s) 16 55.7 3.87 (s) Standard: TMS
¹ H NMR: 600 MHz
¹³ C NMR: 150 MHz

1 is a ¹³ C NMR spectrum of the CrytoSybin A compound according to the present invention, FIG. 2 is a ¹ H NMR spectrum of the CrytoSybin A compound according to the present invention, FIG. The HMBC spectrum of the empty A compound is shown.

FIG. 4 is a ¹³ C NMR spectrum of the CrytoSybin B compound according to the present invention, and FIG. 5 shows a ¹ H NMR spectrum of the CrytoSybin B compound according to the present invention.

FIG. 6 is a ¹³ C NMR spectrum of the CrytoSybin C compound according to the present invention, and FIG. 7 shows a ¹ H NMR spectrum of the CrytoSybin C compound according to the present invention.

As shown in FIGS . 1 to 7 and Table 2, in ¹ H-NMR, hydrogen peaks bound to an aromatic ring are δ6.50 (1H), δ6.71 (1H), and δ6.84. (2H) and δ 7.54 (2H) were observed, and it can be seen that the chemical shift of δ4.71 (2H) was associated with nitrogen. In addition, ¹³ C-NMR and HMQC analysis confirmed the presence of hydroxy carbons of δ 155 (C 1), δ 162 (C 3) and δ 157 (C 12), and of δ 50 (C 8) carbon and δ 172 (C 7). Carbonyl carbon was identified, and meth120 (C11), δ124 (C10, C14), δ124 (C10, C14), δ117 (C11, C13) methine carbon) was observed. In the HMBC spectrum, long range correlations were observed at δ4.71, δ6.50, δ6.71, δ6.84 and δ7.54.

< Experimental Example  1> New Crytocybin Derivatives Radical  Scavenging activity measurement

In order to determine the radical scavenging activity of the novel crytocycline derivative according to the present invention, the following experiment was performed.

(1) 2,2- Azino - Vis-3 - Ethyl Benzoyl Algin Sulfonic Acid ( ABTS ⁺ ) Radical  Scavenging activity measurement

A 7 mM ABTS aqueous solution was mixed with 2.45 mM potassium persulfate to generate ABTS ⁺ radicals, which were then diluted to have an absorbance of about 0.7 at 734 nm to form an ABTS ⁺ aqueous solution. Subsequently, in 190 μl of the prepared ABTS ⁺ radical aqueous solution, a novel crytocybin derivative (Crytocybin A, B, C compound) according to the present invention, a sample such as trolox (trolox), BHA, etc. as a comparison group was DMSO. 10 μl of the solution dissolved in the solution was mixed and left for 7 minutes, and then absorbance was measured at 734 nm. Next, the absorbance of the control group in which only DMSO was added to the ABTS ⁺ radical aqueous solution was measured. Using the measured absorbance, ABTS ⁺ radical scavenging activity was calculated according to Equation 1 below.

(2) Determination of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity

In a 90 μl solution of 150 μM of 1,1-diphenyl-2-picrylhydrazyl (DPPH) dissolved in ethanol, a novel chrythocybin derivative according to the present invention Bin A, B, C compounds), 10 μl of a solution of trolox, BHA and the like as a comparative group in DMSO were mixed and left for 30 minutes, and then absorbance was measured at 517 nm. Next, the absorbance of the control group in which only DMSO was added without adding a sample to the DPPH radical solution was measured. Using the measured absorbance, DPPH radical scavenging activity was calculated according to Equation 2 below.

(3) Determination of Superoxide Radical Scavenging Activity

50 mM potassium phosphate buffer (pH 7.8), 1 mM EDTA, 0.04 mM nitroblue tetrazolium (NBT), 0.18 mM xanthine, 250 mU / ml per well of a 96-well plate A sample solution of xanthine oxidase and a novel crytocybin derivative (Crytocybin A, B, C compound) according to the present invention dissolved in DMSO is added to a total of 100% by weight. The reaction was carried out in the dark at 37 ° C. for 30 minutes. Xanthine oxidase promotes oxidation of xanthine to uric acid and superoxide, and superoxide reduces NBT to form blue formazan. The blue formazan formed by reduction of NBT was measured at 560 nm by ELISA. As a control, only DMSO was added without adding a sample, and ferulic acid or catechin was used as a comparative group. Scavenging activity was calculated according to Equation 2 below using the absorbance difference between the control and test samples. The experiment was performed three times, and the absorbance change was determined by averaging.

% Radical scavenging activity = (ΔA _{control /} ΔA _sample −1) × 100

% Radical scavenging activity = (1-ΔA _sample / ΔA _control ) × 100

(In Equation 1 or 2,

ΔA _sample : Change in absorbance of the well containing the test compound.

ΔA _control : Absorbance change of wells containing DMSO in place of test compound.)

Table 3 shows the measurement results of the radical scavenging activity. The results are shown as concentrations 50% scavenging radicals (EC ₅₀ ).

sample Radical scavenging activity (EC ₅₀ ± SD (μM)) ABTS ⁺ DPPH Superoxide New Crythrocybin A Compounds 6.45 ± 0.32 ＞ 100 10.3 ± 0.43 New Crythrocybin B Compounds 9.63 ± 0.22 ＞ 100 5.98 ± 0.33 New Crythrocybin C Compounds 94.91 ± 5.52 ＞ 100 3.61 ± 0.38 Trolox 14.54 ± 0.08 37.61 ± 1.68 BHA 11.70 ± 0.26 97.09 ± 05.52 Ferulic acid 150.22 ± 6.55 Catechin 47.61 ± 1.68

As shown in Table 3, the radical scavenging activity of the novel crytocycline derivatives according to the invention is 6.45 ± 0.32 μM and 10.3 ± 0.43, respectively, for the novel cryotocycline A compounds in the ABTS ⁺ radicals and the superoxide radicals. In the case of μM and the new Crythrocycline B compound, 9.63 ± 0.22 μM and 5.98 ± 0.33 μM, respectively, were shown to be excellent in radical scavenging activity (trolox, BHA, It can be seen that the radical scavenging activity is far superior to ferulic acid, catechin, and the like. New Crythrocybin C compound also shows 3.61 ± 0.38 μM in the superoxide radical, it can be seen that the radical scavenging activity is superior to the comparative group. As for DPPH radicals, they showed a degree equivalent to that of BHA.

Therefore, the novel crytocybin derivatives according to the present invention exhibit excellent radical scavenging activity, and thus may be usefully used for preventing and aging as well as for preventing and treating diseases caused by excessive accumulation of active oxygen.

Experimental Example 2 Inhibition of Intracellular DNA Damage of Novel Crythrocybin Derivatives

In order to investigate the effect of the novel crytocybin derivatives according to the present invention on reducing cellular DNA damage, a comet assay (Korean Nutrition Journal 37 (6): 440-447, 2004) Was used.

Human lymphocyte cells collected from the blood of a healthy adult woman were treated with no treatment as a control, or 100 μM of novel crytocybin derivatives (Crytocybin A, B, C compounds), hydrogen peroxide (H ₂ O ₂ ), or The amount of tail DNA produced by the addition of a new crytocybin derivative plus hydrogen peroxide was measured to investigate the degree of DNA damage, and image analysis was performed using a fluorescence microscope (Leica DMLB, Germany).

As the production amount of the tail DNA increases, DNA damage increases.

The measurement results are shown in FIG. 8 and Table 4.

In FIG. 8 , (a) is a control group, (b) is a novel crytocybin A compound treatment group according to the present invention, (c) is a novel crytocybin B compound treatment group according to the present invention, and (d ) Is a novel crytocybin C compound treatment group according to the present invention, (e) is a hydrogen peroxide treatment group, (f) is a new crytocybin A compound + hydrogen peroxide treatment group, (g) is a new cryto Cybin B compound + hydrogen peroxide treatment group, (h) is a novel Crythrocybin C compound + hydrogen peroxide treatment group.

tail DNA Average Standard Deviation error Control 5.72 5.10 0.72 New Crythrocybin A Compounds 7.01 5.29 0.75 New Crythrocybin B Compounds 6.49 5.68 0.80 New Crythrocybin C Compounds 5.09 4.18 0.59 Hydrogen peroxide 61.11 17.26 2.44 New Crythrocybin A Compound + Hydrogen Peroxide 35.39 15.92 2.25 New Crythrocybin B Compound + Hydrogen Peroxide 38.34 18.00 2.55 New Crythrocybin C Compound + Hydrogen Peroxide 32.32 15.59 2.25

As shown in Table 4, the treatment of hydrogen peroxide produces an average of 61.11 of tail DNA, but the treatment of cryocybin derivatives according to the present invention produces 32.32-38.34 of tail DNA, which damages about 40% of tail DNA. It can be seen that.

In addition, as a result of photographing a comet analysis image, as shown in FIG . 8 , when hydrogen peroxide (H ₂ O ₂ ) is treated, tails are formed by many tail DNAs (e). The tail of tail DNA was reduced enough to be observed (f ~ h).

Therefore, the cryocybin derivative according to the present invention can be usefully used in anti-aging cosmetic compositions because it reduces the aging promotion by reducing the DNA damage by strong reactive oxygen such as hydrogen peroxide by about 40%.

Experimental Example 3 Ultraviolet Protection Effect of New Crythocybin Derivatives

In order to examine the UV-blocking effect of the novel crytocybin derivatives according to the present invention by light irradiation, experiments were performed using the Guinea Pig Erythema model as follows.

That is, after depilating the albino guinea pig, after applying the novel crytocybin derivative (Crytocybin A, B, C compound) according to the present invention 4 days before, 2 days before, 2 hours before ultraviolet irradiation, immediately after ultraviolet irradiation The effect of UV light on erythema was investigated. Ultraviolet light throughput was investigated under 2MED-400 MJ / cm ² conditions. As a control, PG / EtOH / DIW mixed solution (PG: EtOH: DIW = 5: 3: 2), which is a known antioxidant active substance, was treated.

The erythema was measured by visual observation, photographs, chromameters, and tissue staining. The erythema reduction was calculated according to Equation 3 below.

Erythema reduction (AU) = [(A control -A substance treatment group) / A control group]

* A is the chromameter value.

The measurement results are shown in FIG. 9 .

9 is a novel crytocybin derivative (a ~ c) according to the present invention and It is a graph showing the degree of erythema reduction of the control group (d) known to have excellent erythema reduction effect.

As shown in Figure 5 , the novel crytocybin derivative according to the present invention showed a result of reducing the erythema similar to the control group known to be excellent in erythema reducing effect against ultraviolet irradiation. In addition, the histological changes observed by H & E tissue staining did not show any difference (not shown).

Therefore, the novel crytosaibin derivative according to the present invention can be usefully used as a functional cosmetic composition because it is excellent in the UV blocking effect by light irradiation.

Experimental Example 4 Acute Toxicity Test of Novel Crythrocybin Derivatives

In order to confirm the acute toxicity of the novel crytocybin derivatives according to the present invention, an acute toxicity test was performed by the following method.

Six-week-old SPF rats were used as experimental animals, and five rats were divided per group. The novel crytocybin derivatives prepared in the above examples were each suspended in 0.5% methylcellulose solution and administered orally at a dose of 1 g / kg / 15 ml.

After administration of the test substance, mortality, clinical symptoms, and changes in body weight were observed. Hematological and hematological examinations were performed. Necropsy was performed to observe abdominal and thoracic organ abnormalities.

As a result, all animals treated with test substance showed no clinical symptoms and no dead animals, and no toxicity change was observed in weight change, blood test, blood biochemical test, autopsy findings.

As a result, all of the tested compounds did not show toxic changes up to 1,000 mg / kg in rats, and the minimum lethal dose (LD ₅₀ ) was determined to be a safe substance of at least 1,000 mg / kg or more.

Examples of formulations for the composition of the present invention are illustrated below.

Preparation Example 1 Preparation of Pharmaceutical Formulation

1. Preparation of Tablets

Tablets containing the novel chrythocybin derivative or pharmaceutically acceptable salt of the present invention as an active ingredient were prepared by the following method.

The compounds of the above examples were sieved and mixed with lactose, starch and pregelatinized corn starch, then a suitable volume of purified water was added and granulated into a powder. The granules were dried and then mixed with magnesium stearate and compressed to obtain tablets.

The components of the tablet are as follows.

New Crythrocybin Derivative 5.0 mg

Lactose BP 150.0 mg

Starch BP 30.0 mg

Pregelatinized Corn Starch BP 15.0 mg

1.0 mg magnesium stearate

2. Preparation of Capsule

Capsules containing the novel chrytocybin derivative or pharmaceutically acceptable salt of the present invention as an active ingredient were prepared by the following method.

The compound of the above example was mixed with an amount of excipient and magnesium stearate. The resulting mixture was filled into gelatin capsules to obtain capsules.

The components of the capsule are as follows.

New Crythrocybin Derivative 5.0 mg

Starch 1500 100.0 mg

Magnesium Stearate BP 1.0 mg

3. Preparation of Injection

Injectable solutions containing the novel crytocybin derivatives or pharmaceutically acceptable salts of the present invention as active ingredients were prepared by the following method.

Dissolve the novel chrytocybin derivative in an appropriate volume of sodium chloride BP for injection, adjust the pH of the resulting solution to pH 3.5 with dilute hydrochloric acid BP, and then adjust the volume with sodium chloride BP for injection and sufficiently Mixed. The solution was filled into a 5 ml Type I ampoule made of clear glass, and the glass was dissolved, sealed under an upper grid of air, and then sterilized by autoclaving at 120 ° C. for at least 15 minutes to obtain an injection solution.

The components of the injection solution are as follows.

100 μg / ml of novel crytocybin derivatives

Dilute hydrochloric acid BP until pH 3.5

Injectable sodium chloride BP up to 1 ml

Preparation Example 2 Preparation of External Skin Agent

1. Preparation of Cream

Cetostearyl alcohol 2.8 parts by weight

Milnan 2.6 parts by weight

Stearic acid 1.4 parts by weight

2 parts by weight of glycerol phosphate monostearate

Fiji-100 Stearate 1 part by weight

Sesquioleate sorbitan 1.4 parts by weight

4 parts by weight of jojoba oil

Squalane 3.8 parts by weight

Polysorbate 60 1.1 parts by weight

2 parts by weight of macadamia oil

Tocopherol acetate 0.2 parts by weight

0.4 parts by weight of methylpolysiloxane

0.1 parts by weight of ethyl paraben

0.1 parts by weight of propylparaben

Euxyl K-400 0.1 part by weight

1,3-butylene glycol 7 parts by weight

Methyl paraben 0.05 parts by weight

Glycerin 6 parts by weight

0.2 parts by weight of d-pandenol

4.6 parts by weight of new crytocybin derivatives

0.2 parts by weight of triethanolamine

pt 41891 0.2 parts by weight

pH ₂ O 46.05 parts by weight

2. Preparation of Lotion

Cetostearyl alcohol 1.6 parts by weight

Stearic acid 1.4 parts by weight

Lipophilic monostearic acid 1.8 parts by weight

Fiji-100 Stearate 2.6 parts by weight

0.6 parts by weight of sesquioleate sorbate

4.8 parts by weight of squalene

2 parts by weight of macadamia oil

2 parts by weight of jojoba oil

Tocopherol Acetate 0.4 parts by weight

0.2 parts by weight of methylpolysiloxane

0.1 parts by weight of ethyl paraben

0.1 parts by weight of propylparaben

1,3-butylene glycol 4 parts by weight

0.1 parts by weight of methyl paraben

Xanthan Gum 0.1 parts by weight

Glycerin 4 parts by weight

0.15 parts by weight of d-pandenol

Allantoin 0.1 part by weight

3.5 parts by weight of new crytocybin derivatives

4 parts by weight of carbone (2% aq.Sol)

0.15 parts by weight of triethanolamine

3 parts by weight of ethanol

pt 41891 0.1 parts by weight

pH ₂ 0 48.3 parts by weight

Figure 1 is a ¹³ C NMR spectrum showing a novel crytocybin A compound according to the present invention.

Figure 2 is a ¹ H NMR spectrum showing a novel crytocybin A compound according to the present invention.

Figure 3 is an HMBC spectrum showing a novel crytocybin A compound according to the present invention.

Figure 4 is a ¹³ C NMR spectrum showing a novel crytocybin B compound according to the present invention.

Figure 5 is a ¹ H NMR spectrum showing a novel crytocybin B compound according to the present invention.

Figure 6 is a ¹³ C NMR spectrum showing a novel crytocybin C compound according to the present invention.

Figure 7 is a ¹ H NMR spectrum showing a novel crytocybin C compound according to the present invention.

FIG. 8 is a comet analysis image photograph showing the degree of DNA damage during treatment with a novel crytocybin derivative and hydrogen peroxide according to an embodiment of the present invention ((a) control; (b) a novel crytocybin A compound treatment group; ( c) New Crythrocybin B Compound Treatment Group; (d) New Crythrocybin C Compound Treatment Group; (e) Hydrogen Peroxide Treatment Group; (f) New Crythrocybin A Compound + Hydrogen Peroxide Treatment Group; (g) New Crythrocybin B Compound + Hydrogen Peroxide Treatment Group, and (h) New Crythrocybin C Compound + Hydrogen Peroxide Treatment Group).

Figure 9 is a graph showing the degree of erythema reduction in the treatment of novel cryocybin derivatives according to an embodiment of the present invention ((a) a new cryocybin A compound treatment group; (b) a novel cryocybin B compound (C) a novel Crythrocybin C compound treatment group; and (d) a control group.

Claims (14)

A sunscreen cosmetic composition containing a novel crytocycline derivative represented by Formula 1 as an active ingredient: [Formula 1]

. (In Formula 1, R ¹ and R ² are independently or optionally OCH ₃ or OH.) [Claim 2] The cosmetic composition for sunscreen according to claim 1, wherein the crytocycline derivative is a compound represented by the following Chemical Formula 1a, 1b or 1c: [Formula 1a]

[Chemical Formula 1b]

[Formula 1c]

. Obtaining a culture solution by culturing the hygrophoropsis aurantiaca KCTC 11143BP strain (Step 1); Extracting the culture solution obtained in step 1 with acetone and ethyl acetate (step 2); Obtaining an active fraction by performing concentration gradient silica gel column chromatography on the extract extracted in step 2 using a mixed solvent of chloroform and methanol (step 3); And The novel cryotope of claim 1 comprising the step (step 4) of the active fraction obtained in step 3 using methanol (100%) as a solvent to perform Sephadex LH-20 column chromatography to obtain a compound of formula 1 Method for preparing cyvin derivatives. The method according to claim 3, wherein the step 1 comprises the culturing of the novel crytocybin derivative of claim 1, wherein the strain of Hygrophoropsis aurantiaca KCTC 11143BP is cultured in a yeast extract (YPS) liquid medium. Manufacturing method. The method of claim 3, wherein the concentration gradient of the mixed solvent of chloroform and methanol in step 3 is 50: 1 to 1: 1. delete delete delete delete delete delete delete delete delete

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