KR101620815B1 - Composition for the prevention and treatment of inflammatory diseases comprising the penicillinolide A isolated from marine fungi - Google Patents

Abstract

The present invention relates to a pharmaceutical composition comprising Penicillium The present invention relates to a novel penicillinolide A compound having anti-inflammatory activity isolated from a strain SF-5292 and its use, wherein the penicillinolide A isolated from marine fungi of the present invention is a compound of the formula Cain of IL-1β, TNF-α, IL-6, effectively inhibited the generation and activation of NO and PGE _2, and suppresses the NO release of which can lead to inflammation directly effectively, the inflammation induced protein expression inhibitory substance It inhibits migration into the nucleus, increases the expression and activity of HO-1, an enzyme known to exhibit cytoprotective effects from oxidative stress, and has low cytotoxicity due to natural products, resulting in safety and adverse side effects , The compounds of the present invention can be usefully used as a composition for the prevention or treatment of inflammatory diseases.

Description

TECHNICAL FIELD The present invention relates to a composition for preventing and treating inflammatory diseases, which comprises penicillinolide A isolated from marine fungi as an active ingredient, and a composition for preventing and treating inflammatory diseases comprising a penicillinolide A isolated from marine fungi

The present invention relates to a novel compound isolated from marine fungi, its use, and a method for separating said compound.

Inflammation is a localized immune response that minimizes the damage of a cell or tissue when it is damaged or destroyed by various factors, such as harmful substances or organisms from the outside, and restores the damaged area to the original state. And is a useful defense mechanism to remove the products produced by tissue damage. As a result, pain, swelling, redness, fever, or the like may occur as a result of the defensive mechanism, resulting in malfunction. Various factors causing inflammation include physical factors such as trauma, burn, bronze, radioactivity, chemical factors such as acid, and immunological factors due to antibody reaction. In addition, blood vessel and hormone imbalance .

The inflammation normally functions to neutralize or eliminate the causative factor through in vivo inflammatory reaction and regenerate the upper tissue to restore the normal structure and function. However, when the degree of inflammation is over a certain level or becomes chronic, chronic inflammation And the like. In addition to being able to observe inflammatory responses in almost all diseases of clinical diseases, enzymes related to inflammatory reaction play an important role in carcinogenesis.

Recently, it has been known that the progress of the inflammatory reaction in the body is related to COX (cyclooxygenase) enzyme activity. The COX enzyme is a major enzyme involved in the biosynthesis of prostaglandin in vivo (Smith et al., J. Biol. Chem., 271, 33157 (1996)), and two kinds of isozyme enzymes, COX-1 and COX -2 is known to exist. The COX-1 is constantly present in tissues such as the stomach or kidney and is involved in maintaining normal homeostasis, while the COX-2 is involved in mitogen or cytokines in inflammation or other immune responses. Is a transient and rapidly expressed enzyme in the cell.

Another strong inflammatory mediator, nitric oxide (NO), is produced from L-arginine by NO synthase (NOS), and is produced by a variety of substances, such as UV, external stress, endotoxin or cytokines Lt; / RTI > cells. These inflammatory stimuli increase the expression of inducible NOS (iNOS) in the cells, thereby inducing NO production in the cells and activating the macrophages to cause an inflammatory reaction.

In addition, the expression or activity of Heme Oxygenase-1 (HO-1), an enzyme closely related to the cell protection mechanism, is closely related to antioxidant and anti-inflammatory mechanisms .

The major function of the Heme Oxygenase-1 (HO-1) is to induce the oxidation of Heme to convert the Heme into biliverdin, free iron ) And carbon monoxide. The bovine peroxidase is converted to bilirubin by biliverdin reductase, which has been reported to exhibit antioxidative effects by acting as a reactive oxygen species (ROS) scavenger.

Particularly, carbon monoxide generated by the hematin oxidase-1 has an anti-inflammatory effect, and the free iron is involved in ferritin synthesis and has a cytoprotective effect. By this action, the hematinoxidase-1 (HO-1) is able to induce oxidation of the hematin to form biliverdin, ultimately bilirubin, free iron, It produces decomposition products of carbon monoxide, exerts cytoprotective effect against external stimulus, and has anti-inflammatory effect.

Therefore, it has been reported that the increase in the expression or activity of hematinoxidase-1 has important cellular mechanisms in relation to anti-inflammatory actions as well as cell protection against external stimuli.

Therefore, in order to effectively alleviate inflammation, researches on substances capable of inhibiting the production of NO and expressing or activating hematin oxidase-1 have been conducted.

Recent researches on the search and use of natural products have explored and used the biosynthesis ability of organisms. So far, mainly plants and microorganisms have been the major species of the object. In particular, microorganisms have a variety of species and produce unique and interesting physiologically active substances, have a short generation period, are easy to mass-produce, and have high industrial availability, and thus have become attractive sources of bioactive materials. Many of the physiologically active substances found from microorganisms originate from actinomycetes, fungi, and bacteria, and about 25% of them are found as fungi, which are very important microbial resources in industrial or medical terms It is known.

Accordingly, the present inventors have searched for a substance Penicillium sp. SF-5292 producing a substance having excellent anti-inflammatory activity from marine invertebrates while searching for a substance having excellent anti-inflammatory activity from a metabolite of a marine microorganism From which novel structured penicillinolide A compounds were isolated and purified purely, and these compounds were tested for their ability to inhibit the inflammatory cytokines IL-1β, TNF-α, IL-6, NO and PGE ₂ Effective inhibition of the production and activation of NO, capable of directly inhibiting the secretion of NO that can directly induce inflammation, inhibiting the migration of an inhibitor of inflammation-inducing protein expression into the nucleus, protecting cells from damage due to oxidative stress In addition to increasing the expression and activity of HO-1, an enzyme known to be effective, it is also a substance derived from natural materials and has low cytotoxicity, And the possibility of occurrence of a problem is low.

It is an object of the present invention to provide a novel compound isolated from marine fungi, its use, and a method for separating said compound.

In order to achieve the above object, the present invention provides a compound represented by the following formula (1), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

The present invention also provides a penicillinolide A compound represented by the following formula (2) or a pharmaceutically acceptable salt thereof:

The present invention also relates to a method for producing a microorganism which comprises 1) culturing a strain of Penicillium sp.

2) extracting the culture obtained in step 1) with methyl ethyl ketone; And

3) separating the methyl ethyl ketone extract obtained in the step 2) by column chromatography.

The present invention also provides a strain of the genus Penicillium deposited under Accession No. KCTC 18284P.

The present invention also provides a pharmaceutical composition for the prevention and treatment of inflammatory diseases containing the compound represented by the general formula (2), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof as an active ingredient.

The present invention also provides a health functional food for preventing and / or improving inflammatory diseases, which comprises the compound represented by the general formula (2), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof as an active ingredient.

The novel penicillinolide A compound of the present invention effectively inhibits the production and activation of IL-1β, TNF-α, IL-6, NO and PGE ₂ , which are inflammation-related cytokines, The expression and activity of HO-1, which is known as an enzyme that effectively inhibits the secretion of NO, inhibits the migration of an inhibitor of inflammation-inducing protein expression into the nucleus, and exhibits a cytoprotective effect from damage due to oxidative stress And since the possibility of causing safety and adverse reaction problems is low as a natural material, penicillinolide A can be usefully used as a composition for the prevention or treatment of inflammatory diseases.

1 shows a ¹ H NMR spectrum (400 MHz, pyridine- d ₆ ) of a compound (penicillinolide A) isolated in the present invention.
FIG. 2 is a graph showing a ¹³ C NMR spectrum (100 MHz, pyridine- d ₆ ) of the compound (penicillinolide A) isolated in the present invention.
FIG. 3 shows the results of confirming the expression changes of inflammation-related substances (IL-1β, TNF-α, IL-6, NO and PGE ₂ ) upon treatment with the compound (penicillinolide A) The graph is:
(A) is a graph showing the results of measuring the content of IL-1β, (B) is a graph showing the results of measuring the content of TNF-α, and (C) (D) is a graph showing the results of measuring the content of NO, and (E) is a graph showing the results of measuring the content of PGE ₂ .
In the graph, '-' means no treatment, '+' means treatment, numerical value means the throughput (μM) of penicilanolide A, and the remaining indication means that the treated sample , And the vertical axis of the graph represents the content of the inflammation-related substance to be analyzed.
FIG. 4 is a graph showing the results of Western blot analysis of iNOS protein expression and COX-2 protein expression level when the compound (penicillinolide A) isolated in the present invention was treated.
(A) is a photograph showing the amount of protein expression, (B) is a graph of iNOS among the results of (A), and (C) will be.
In the photograph, '-' means no treatment, '+' means treatment, numerical value means the throughput (μM) of penicillinolide A, and the remaining indication means the treated sample or assay ≪ / RTI >
FIG. 5 is a graph showing the content and activation level associated with IκB-α and NF-κB upon treatment of the compound (penicillinolide A) isolated in the present invention:
(A) and (B) show the content of IκB-α and phosphorylated IκB-α (phosphorylated IκB-α, p-IκB-α) in the cytoplasmic fraction and the content of p65 in the nuclear fraction, (C) is a graph showing the binding activity of NF-κB in the nuclear fraction.
The numerical value of the photograph means the throughput (μM) of penicillanolide A, p-IκB-α means phosphorylated IκB-α (phosphorylated IκB-α), p65 means NF-κB, 'Means no treatment,' + 'means treatment, numerical value means the throughput (μM) of penicillinolide A, and the vertical axis indicates NF -kB < / RTI > as a relative value.
FIG. 6 is a diagram showing the amount of hemine oxidase-1 produced when the compound (penicillinolide A) isolated in the present invention is treated. FIG.
(A) and (B) show the degree of production of HO-1 according to the concentration of penicillinolide A in terms of concentration (horizontal axis, μM) or whether or not CoPP was used in the control group (C) shows the effect of inducing HO-1 expression by penicillinolide A according to the time (horizontal axis, hour (h)) of treatment with 40 μM of penicilinoid A And the degree of HO-1 production in an untreated control (control).
The values in (A) and (B) refer to the amount of penicillanolide A, and the value in (C) means the time (h) after the treatment with penicillinolide A.

Hereinafter, the present invention will be described in detail.

The present invention provides a compound represented by the following formula (1), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

[Chemical Formula 1]

.

.

The compound is characterized in that it is isolated from the Penicillinolide A strain represented by the formula (2)

(2)

.

.

The compound is preferably isolated from the genus Penicillium sp., And is more preferably isolated from the genus Penicillium SF-5292 deposited with Accession No. KCTC 18284P.

In a specific example of the present invention, the present inventors isolated a strain from a marine invertebrate sample, separated the compound from the separated strain of Penicillium genus SF-5292, analyzed the structure thereof, named it penicillinolide A Its structure was analyzed (see Figs. 1 to 2).

The present invention includes all the possible solvates and hydrates, which can be prepared therefrom, as well as the compound represented by the formula (2) or a pharmaceutically acceptable salt thereof.

The compound of formula (2) of the present invention can 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 innocuous salts include, but are not limited to, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate chloride, bromide, Butyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, succinate, maleic anhydride, maleic anhydride, , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzene sulfonate, toluene sulfonate, chlorobenzene sulfide Propyl sulphonate, naphthalene-1-yne, xylenesulfonate, phenylsulfate, phenylbutyrate, citrate, lactate,? -Hydroxybutyrate, glycolate, maleate, Sulfonate, naphthalene-2-sulfonate or mandelate.

The acid addition salt according to the present invention can be produced by a conventional method, for example, by dissolving the compound of formula (1) in an excess amount of an acid aqueous solution, and using this salt in a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile Followed by precipitation.

By heating the same amount of the compound of formula (II) and an acid or alcohol in water, and then evaporating and drying the mixture, or by suction filtration of the precipitated salt.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess amount of an alkali metal hydroxide or alkaline earth metal hydroxide 2 solution, filtering the insoluble compound salt, and evaporating the filtrate and drying. At this time, it is preferable for the metal salt to produce sodium, potassium or calcium salt. In addition, the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

In addition,

1) culturing a strain of the genus Penicillium;

2) extracting the culture obtained in step 1) with methylethylketone; And

3) separating the methyl ethyl ketone extract obtained in the step 2) by column chromatography. The present invention also provides a method for separating a penicillinolide A compound of the present invention.

In this method, step 1) is preferably a step of culturing a strain of the genus Penicillium, wherein the culturing of the strain is preferably carried out in a culture medium containing a nutrient source to which conventional microorganisms can be used. As the nutrient source, it is possible to use a known nutrient source used for culturing the mold. For example, the carbon source may be glucose, starch syrup, dextrin, starch, molasses, animal oil, vegetable oil, etc. The nitrogen sources may include wheat bran, soybean meal, wheat, malt, cottonseed oil, Extracts, ammonium sulfate, sodium nitrate, urea and the like can be used. If necessary, addition of inorganic salts promoting salt formation, potassium, magnesium, cobalt, chlorine, phosphoric acid, sulfuric acid and other ions is very effective. The culture method is preferably a shaking culture or a stationary culture under aerobic conditions, but is not limited thereto.

The strain of the genus Penicillium in step 1) above is preferably a strain of Penicillium genus SF-5292 deposited under accession number KCTC 18284P.

The culture temperature is slightly different depending on the conditions when culturing under the above-mentioned conditions, but it is preferably cultured at 20 to 37 ° C, more preferably at 23 to 30 ° C, but is not limited thereto.

In the above method, the step 2) is a step of extracting the culture of the strain obtained in the step 1) with methyl ethyl ketone, and the penicilanolide A compound of the present invention may be present in the mycelium as well as the culture medium of the strain. Therefore, it is preferable to add an organic solvent such as methyl ethyl ketone to the culture broth and mycelia of the strain, extract the active ingredient from the culture broth and the mycelium, and concentrate the obtained extract by the reduced pressure evaporation method.

In the above method, Step 3) is a step of isolating the penicillanolide A compound of the present invention, wherein the methyl ethyl ketone concentrate obtained in Step 2) is subjected to flash column chromatography using a mixed solvent of methanol: dichloromethane Followed by purification by high performance liquid chromatography and separation.

The present invention also provides a strain of the genus Penicillium deposited under Accession No. KCTC 18284P.

After isolating the strain from marine invertebrates, the present inventors have identified 28S rRNA sequence analysis of the strain as Penicillium genus having the nucleotide sequence of SEQ ID NO: 1. In the present invention, the strain is treated with Penicillium sp.) SF-5292, deposited with KCTC in the Korea Research Institute of Bioscience and Biotechnology (KCTC) on April 25, 2014 (accession number: KCTC 18284P).

The present invention also provides a pharmaceutical composition for the prevention and treatment of inflammatory diseases containing the compound of the present invention, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof as an active ingredient.

The compound is preferably isolated from a marine invertebrate sample and is preferably isolated from a strain belonging to the genus Penicillium, but is not limited thereto.

Wherein said strain belongs to the genus Penicillium SF-5292 deposited with Accession No. KCTC 18284P.

The compound is characterized by inhibiting the production of IL-1β, TNF-α, IL-6, NO and PGE _2.

In the present specification, inflammation refers to a general inflammatory disease, and the inflammatory disease is any one selected from the group consisting of dermatitis, allergy, systemic lupus erythematosus, retinitis, gastritis, hepatitis, enteritis, pancreatitis, .

More specifically, the inflammatory diseases are various acute inflammatory diseases such as dermatitis, allergies, ophthalmic diseases such as systemic lupus erythematosus, retinitis, various chronic inflammatory diseases such as rheumatoid arthritis, lupus, multiple sclerosis, sepsis, , Bronchitis, inflammatory bowel disease such as gastritis or enteritis, hepatitis, pancreatitis, nephritis, and combinations thereof.

In a specific example of the present invention, the anti-inflammatory effect of the compound of the present invention, penicillinolide A, was confirmed. As a result, IL-1β, TNF-α, IL-6, NO (nitric oxide, nitric oxide) (COX-2 (cyclooxygenase 2) inducing PGE ₂ production and iNOS inducible nitric oxide synthase (PGE 2) inducing NO production by inhibiting concentration of PGE ₂ (prostaglandin 2, prostaglandin E 2) As a result, it was confirmed that the expression of iNOS and COX-2 protein decreased in a concentration-dependent manner when penicillinolide A was added (see FIG. 4), and NF-κB (p65) inhibited phosphorylation and degradation of IκB-α, a substance that inhibits nuclear translocation. As a result, the expression of IκB-α was increased in a concentration-dependent manner when penicillinolide A was treated, Induced by Inhibition of IκB-α was inhibited and the expression of p-IκB-α was decreased in a concentration-dependent manner. Thus, the inhibition of IκB-α activation induced by LPS was inhibited, and the expression of p65 was decreased in a concentration- Induced NF-κB translocation into the nucleus was inhibited and the degree of DNA binding of NF-κB was decreased in a concentration-dependent manner (see FIG. 5).

In addition, the expression and activity of hematoxylin-1 (Heme Oxygenase-1, HO-1), known as an inflammation-related enzyme, were measured. As a result, the activity of HO- , And the amount of HO-1 produced increased in a concentration-dependent manner. It was confirmed that the amount of HO-1 produced increased with the lapse of the time of penicillinolide A treatment (see FIG. 6).

Thus, the penicillinolide A isolated from the marine fungi of the present invention effectively inhibits the production and activation of IL-1β, TNF-α, IL-6, NO and PGE ₂ , which are cytokines related to inflammation, , The expression of HO-1, which is known as an enzyme that inhibits the secretion of NO, which induces the induction of inflammation-inducing protein, into the nucleus, and protects against the damage caused by oxidative stress And activity thereof, and it is confirmed that it can be used as a pharmaceutical composition for prevention and treatment of inflammatory diseases because the possibility of side effects is not a problem as it is separated from natural materials.

The compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof may be used alone in the composition, and may further include a pharmacologically acceptable carrier, excipient, diluent or subcomponent. More particularly, when a composition comprising the compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof is used as a medicine or used for pharmaceutical or pharmaceutical purposes, the active ingredient may be administered by a conventional method, May be mixed with an acceptable carrier or excipient or diluted with a diluent.

In this case, the content of the active ingredient in the composition may be 0.001 wt% to 99.9 wt%, 0.1 wt% to 99 wt%, or 1 wt% to 50 wt%, but is not limited thereto, The content of the compound may be appropriately adjusted in a desired amount.

Examples of the pharmaceutically acceptable carrier, excipient or diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, propylhydroxybenzoate, talc, magnesium stearate and mineral oil , Dextrin, calcium carbonate, propylene glycol, liquid paraffin, and physiological saline. However, the present invention is not limited thereto, and any conventional carrier, excipient or diluent may be used. In addition, the pharmaceutical composition may further comprise at least one selected from the group consisting of conventional fillers, extenders, binders, disintegrants, anticoagulants, lubricants, humectants, pH adjusters, nutrients, vitamins, electrolytes, alginic acid and its salts, pectic acid and its salts, , Fragrances, emulsifiers, preservatives, and the like. The components may be added independently or in combination with any one selected from the group consisting of the above-mentioned active ingredient, pellucamine G, a pharmaceutically acceptable salt thereof, and combinations thereof.

When the composition is used as a medicament, the administration method can be either oral or parenteral, and can be administered through various routes including oral, transdermal, subcutaneous, intravenous or muscular.

In addition, the formulations of the compositions may vary depending on the method of use and may be formulated using methods well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal . In general, solid dosage forms for oral administration include tablets, pills, soft or hard capsules, pills, powders and granules, and the like, which may contain one or more Excipients such as starch, calcium carbonate, sucrose or lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration include sustainsions, liquids, emulsions and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, Sweeteners, fragrances, preservatives, and the like.

Forms for parenteral administration include, but are not limited to, creams, lotions, ointments, plasters, liquids and solutions, aerosols, fruidextracts, infusion, elixir, infusions, sachet, patch or injections, and the like.

Furthermore, the compositions of the present invention may be formulated using any suitable method known in the art or using methods disclosed in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA .

The dosage of the composition can be determined in consideration of the administration method, the age, sex, the severity of the patient, the state of the recipient, the degree of absorption of the active ingredient in the body, the inactivation rate and the drug to be used together, Kg body weight to 500 mg / kg body weight, 0.1 mg / kg body weight to 400 mg / kg body weight or 1 mg / kg body weight to 300 mg / kg body weight, And can be administered once or several times in divided doses. The dose is not intended to limit the scope of the invention in any way.

The present invention also provides a health functional food for the prevention and improvement of inflammatory diseases containing the compound of the present invention, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof as an active ingredient.

The compound is preferably isolated from a marine invertebrate sample and is preferably isolated from a strain of the genus Penicillium, but is not limited thereto.

Wherein said strain belongs to the genus Penicillium SF-5292 deposited with Accession No. KCTC 18284P.

The compound is characterized by inhibiting the production of IL-1β, TNF-α, IL-6, NO and PGE _2.

In the present specification, inflammation refers to a general inflammatory disease, and the inflammatory disease is any one selected from the group consisting of dermatitis, allergy, systemic lupus erythematosus, retinitis, gastritis, hepatitis, enteritis, pancreatitis, .

More specifically, the inflammatory diseases are various acute inflammatory diseases such as dermatitis, allergies, ophthalmic diseases such as systemic lupus erythematosus, retinitis, various chronic inflammatory diseases such as rheumatoid arthritis, lupus, multiple sclerosis, sepsis, , Bronchitis, inflammatory bowel disease such as gastritis or enteritis, hepatitis, pancreatitis, nephritis, and combinations thereof.

Phenysilanolide A isolated from marine fungi of the present invention effectively inhibits the production and activation of IL-1β, TNF-α, IL-6, NO and PGE ₂ , which are cytokines related to inflammation, The expression and activity of HO-1, which is known as an enzyme that effectively inhibits the secretion of NO, inhibits the migration of an inhibitor of inflammation-inducing protein expression into the nucleus, and exhibits a cytoprotective effect from damage due to oxidative stress And it is confirmed that it is isolated from natural materials and can be used as an effective ingredient of a health functional food for prevention and improvement of inflammatory diseases because the possibility of side effects is not a problem.

In order for the compound of the present invention to be used for the prevention and improvement of inflammatory diseases as described above, it may be prepared by various methods known in the field of food science or pharmaceuticals, and may be mixed with itself or a pharmaceutically acceptable carrier, excipient, And may be manufactured in any food form that can be taken orally. Preferably in the form of beverage, ring, granule, tablet or capsule.

The health functional food of the present invention may further comprise ingredients that are conventionally added at the time of food production and which are pharmaceutically acceptable. For example, in the case of beverage, one or more components may be further added in addition to the compound of the present invention in citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice and the like.

The amount that can be included as an active ingredient of the health functional food according to the present invention may be appropriately selected according to the age, sex, weight, condition, and symptom of a person who desires to prevent or improve the inflammatory disease, 0.01 g to 10.0 g. It is possible to obtain the effect of preventing and improving inflammatory diseases by ingesting the health functional food having such a content.

The present invention also provides a cosmetic composition for prevention and improvement of inflammatory diseases containing the compound of the present invention, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof as an active ingredient.

The compound is preferably isolated from a marine invertebrate sample and is preferably isolated from a strain of the genus Penicillium, but is not limited thereto.

Wherein said strain belongs to the genus Penicillium SF-5292 deposited with Accession No. KCTC 18284P.

The compound is characterized by inhibiting the production of IL-1β, TNF-α, IL-6, NO and PGE _2.

In the present specification, inflammation refers to a general inflammatory disease, and the inflammatory disease is any one selected from the group consisting of dermatitis, allergy, systemic lupus erythematosus, retinitis, gastritis, hepatitis, enteritis, pancreatitis, .

More specifically, the inflammatory diseases are various acute inflammatory diseases such as dermatitis, allergies, ophthalmic diseases such as systemic lupus erythematosus, retinitis, various chronic inflammatory diseases such as rheumatoid arthritis, lupus, multiple sclerosis, sepsis, , Bronchitis, inflammatory bowel disease such as gastritis or enteritis, hepatitis, pancreatitis, nephritis, and combinations thereof.

Phenysilanolide A isolated from marine fungi of the present invention effectively inhibits the production and activation of IL-1β, TNF-α, IL-6, NO and PGE ₂ , which are cytokines related to inflammation, The expression and activity of HO-1, which is known as an enzyme that effectively inhibits the secretion of NO, inhibits the migration of an inhibitor of inflammation-inducing protein expression into the nucleus, and exhibits a cytoprotective effect from damage due to oxidative stress And it is confirmed that it can be used as an active ingredient of a cosmetic composition for prevention and improvement of inflammatory diseases because it is isolated from a natural product and the possibility of side effects is low.

The cosmetic composition may be in the form of a solution, a gel, a solid or a paste anhydrous product, an emulsion obtained by dispersing an oil phase in water, a suspension, a microemulsion, a microcapsule, a microgranule or an ionic (liposome) A cream, a skin, a lotion, a powder, an ointment, a spray, or a cone stick. It can also be prepared in the form of a foam or an aerosol composition further containing a compressed propellant.

The cosmetic composition may further contain, in addition to the compound of the present invention, a pharmaceutically acceptable salt thereof or a stereoisomer thereof, a fatty substance, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, a softening agent, an antioxidant, a suspending agent, A foaming agent, a fragrance, a surfactant, water, an ionic or nonionic emulsifier, a filler, a sequestering and chelating agent, a preservative, a vitamin, a barrier, a wetting agent, Lipophilic active agents, lipid vesicles, or any other ingredients commonly used in cosmetics.

In a cosmetic composition containing a compound of the present invention, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, the compound of the present invention, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof is added to a cosmetic composition containing 0.005 By weight to 50% by weight.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

< Example  1> Aspergillus  genus( Aspergillus sp .) SF Isolation of -5044 strain

This strain was isolated from marine invertebrates on the coast of Jeju Island in February 2009. The collected samples were placed in a sterilized plastic bag immediately after collection and stored frozen until used in the experiment. Samples for the experiment were diluted 10 times with sterilized seawater and 1 ml of the diluted solution was plated on a potato dextrose agar medium containing 3% NaCl and cultured at 25 ° C for 14 days. The pure strains were separated by several separation steps and the strains were stored at -70 ℃.

< Example  2> Identification and naming of strains

The 28S rRNA sequence was confirmed by rRNA sequencing of this strain.

TTTTTTCTCCTCCCCCCCCCCCCGAAATATCATAAGCGGAGGAAAAGAAACCAACAGGGATTGCCCCAGTAACGGCGAGTGAAGCGGCAAGAGCTCAAATTTGAAAGCTGGCTCCTTCGGGGTCCGCATTGTAATTTGTAGAGGATGCTTCGGGAGCGGTCCCCATCTAAGTGCCCTGGAACGGGACGTCATAGAGGGTGAGAATCCCGTATGGGATGGGGTGTCCGCGCCCGTGTGAAGCTCCTTCGACGAGTCGAGTTGTTTGGGAATGCAGCTCTAAATGGGTGGTAAATTTCATCTAAAGCTAAATATTGGCCGGAGACCGATAGCGCACAAGTAGAGTGATCGAAAGATGAAAAGCACTTTGAAAAGAGAGTTAAAAAGCACGTGAAATTGTTGAAAGGGAAGCGCTTGCGACCAGACTCGCTCGCGGGGTTCAGCCGGCATTCGTGCCGGTGTATTTCCCCGCGGGCGGGCCAGCGTCGGTTTGGGCGGTCGGTCAAAGGCCCTCGGAAGGTAACGCCCCTAGGGGCGTCTTATAGCCGAGGGTGCAATGCGACCTGCCTAGACCGAGGAACGCGCTTCGGCTCGGACGCTGGCATAATGGTCGTAAGCGACCCGTCTTGAAACACGGACCAAGGAGTCTAACATCTACGCGAGTGTTCGGGTGTCAAACCCGTGCGCGAAGTGAAAGCGAACGGAGGTGGGAACCCTTACGGGTGCACCATCGACCGATCCTGAAGTCTTCGGATGGATTTGAGTAAGAGCGTAGCTGTTGGGACCCGAAAGATGGTGAACTATGCCTGAATAGGGCGAAGCCAGAGGAAACTCTGGTGGAGGCTCGTAGCGGTTCTGACGTGCAAATCGATCGTCGAATTTGGGTATAGGGGCGAAAGACTAATCGAACCATCTAGC (SEQ ID NO: 1)

As a result of GenBank search for the nucleotide sequence obtained through 28S rRNA sequence analysis of this strain, Penicillium expansum (JN938952) and showed a homology of 99.89%. The strain was identified as Penicillium spp., Which was identified as Penicillium spp. (KCTC 18284P) deposited on Apr. 25, 2014 with KCTC (Korea Research Institute of Bioscience and Biotechnology) (KCTC).

< Example  3> Penicillium  genus( Penicillium sp .) SF Culture of -5292 strain

In order to cultivate this strain, a medium containing a nutrient source normally used by microorganisms and sodium chloride was prepared. As a crude medium and production medium for the strain, potato starch containing 0.4% (w / v) potato starch, 2% (w / v) dextrose, 3% (w / v) sodium chloride and 1.5% (w / v) Dextrose agar medium was used.

100 ml of the Erlenmeyer flask containing 20 ml of the seed medium was sterilized at 121 ° C for 20 minutes, platinum was inoculated from a slope culture test tube of Aspergillus sp. Strain SF-5044 and cultured for 4 days with shaking Species culture was performed. Two hundred ml of the seed culture medium was inoculated into each of 110 plate culture media made up of 20 ml of the production medium and the cells were cultured at 25 ° C for 10 days.

< Example  4> Penicillium  genus( Penicillium sp .) SF Isolation and purification of compounds from strain -5292

The culture of the Penicillium sp. SF-5292 strain cultured in Example 3 was extracted with 2 L of methylethylketone and the obtained extract was evaporated under reduced pressure using a vacuum dryer And concentrated to obtain 2 g of the extract. The concentrate was adsorbed on Odyssey 18 and subjected to ODS RP-18 flash column chromatography. The methanol / water (2: 8-10 / 0, v / v) And eluted with increasing methanol concentration stepwise as a solvent. At this time, fraction eluted from 80% methanol was subjected to silica gel fresh column chromatography using methanol / dichloromethane (0% -20%, v / v) as a solvent mixture. The fractions eluted with a mixed solvent of methanol / dichloromethane (1:19, v / v) were concentrated under reduced pressure and purified by high performance liquid chromatography (column: Agilent semiprep-C18) using 0.1% formic acid (13.4 mg, t _R = 31.0 min) with a UV absorption peak at 210 nm was eluted with a gradient of 40% to 100% methanol and an elution flow rate of 5 ml / min.

< Example  5> Penicillium  genus SF5292  Structural analysis of compounds from strains

The molecular weight and the molecular formula were determined using an ESI mass spectrometer (ESI mass spectrometer) of the formula (2) isolated in Example 4 above. Further, ¹ H NMR, ¹³ C NMR, COZY (Correlation Spectroscopy), HMQC (1H-Detected heteronuclear multiple-quantum coherence), Heteronuclear Multiple-Bond Coherence (HMBC), and Distortionless Enhancement (DEPT) through nuclear magnetic resonance by Polarization) and NOESY (Nuclear Overhauser effect spectroscopy) spectra, and the molecular structure of the compounds was determined. As a result, Formula (2) was identified as a novel compound containing a large lactone of 10-membered ring derived from a fungus as shown in the following formula. The above formula 2 was named penicillinolide (Fig. 1 to Fig. 2).

(2)

Penicillinolide A (Formula 2): white solid; [[alpha]] _D +50 ( c 0.38, MeOH); FT-IR? Max 3393, 2956, 2932, 2859, 1717, 1440, 1350, 1250, 1128, 1100, 1043, 963, 925 cm @ -1; ¹ H, &lt; ¹³ &gt; C NMR data are shown in Table 1 below; HRESIMS m / z 295.1517 [M + Na] + (calcd for calcd for C 14 H 24 O 5 Na, 295.1521).

NMR data of the formula 2 (pyridine- d ₅ of the measurement) Position δC δH, mult (J in Hz) Key NOESY HMBC (H? C #) One 172.9 - - - 2 28.9 3.11, m 2.56, m - 1, 3, 4 3 28.6 2.65, m 2.00, m - 1, 2, 4, 5 4 65.2 5.00, m H-7 - 5 46.2 3.12, dd (18.1, 11.0) 2.93, dd (18.1, 4.8) - 3, 4, 6 6 211.0 - - - 7 75.2 4.57, dd (7.7, 3.7) H-4, H-9 8, 9 8 39.3 2.58, m 2.41, m - 6, 7, 9, 10 9 73.1 5.27, m H-7 1, 7 10 34.3 1.81, m 1.70, m - 8, 9, 11, 12 11 25.8 1.27, m - 10, 12, 13 12 31.8 1.23-1.09, m - - 13 22.7 1.21-1.11, m - - 14 14.1 0.76, t (6.6) - 12, 13

< Example  6> Penicillinoide  A to inhibit the formation of inflammation-related substances

In order to confirm the antiinflammatory effect of penicillinolide A, the following experiments were carried out to confirm the inhibition of IL-1β, TNF-α, IL-6, NO and PGE ₂ production known as inflammation-related substances.

Specifically, rat and mouse macrophages were treated with 20 and 40 μM of Pulchellamin G and Penicillinolide A (Formula 2), and cultured for 12 hours. Then, LPS (bacterial lipopoly-saccharide) 1 μg / ml, and further cultured for 18 hours to induce an inflammatory response. After incubation, centrifugation was carried out at 13,000 x g and 4 ° C for 2 minutes, and the supernatant was collected.

The secretion amount of TNF-α and IL-1β was measured using a commercially available measurement kit (R & D Systems, USA) according to the protocol provided by the manufacturer. The supernatant of each control and experimental group was added to a 96-well plate coated with mouse-derived TNF-α or IL-1β antibody, and then the secondary antibody (enzyme-linked polyclonal antibody specific for mouse TNF- 1β) was added and reacted for 2 hours, followed by washing to remove unattached secondary antibody. Subsequently, the content of TNF-α or IL-1β was measured by adding the substrate solution and measuring the fluorescence intensity at 450 nm.

The secretion of NO was measured using the Griess reagent system (Promega, USA) according to the protocol provided by Promega. A solution prepared by dissolving 0.1% (w / v) N- (1-naphathyl) -ethylene diamine in 100 μl of the supernatant of each control group and the same amount of Griess reagent (5% (v / v) phosphoric acid solution) 1% (w / v) sulfanil amide) was mixed and allowed to react at room temperature (15 ° C to 20 ° C) for 10 minutes. The absorbance was then measured at 525 nm using an ELISA plate reader.

The secretion amount of PGE ₂ was measured by using commercially available PGE ₂ (R & D Systems, USA) according to the protocol provided by the manufacturer. The supernatant of each control and test group was added to a 96-well plate coated with antibody against PGE ₂ , followed by addition of an enzyme-linked polyclonal antibody specific for PGE ₂ for 20 hours, The secondary antibodies were washed to remove them. Then, the content of PGE ₂ was measured by adding the substrate solution and measuring the fluorescence intensity at 450 nm.

As shown in FIG. 3, IL-1β, TNF-α, IL-6, NO and PGE ₂ , which are known as inflammation-related substances, are low in the control group without addition of LPS, (Fig. 3). When penicillinolide A was added, the amount was decreased in a concentration-dependent manner (Fig. 3).

< Example  7> Penicillinoide  A, anti-inflammatory effects

<7-1> Cellular fraction cytosolic fraction ), And nuclear and cytoplasmic fractions ( Nuclear  and cytoplasmic extract )

In order to confirm the effect of penicillinolide A, which is confirmed to be safe because of its lack of cytotoxicity, on the expression of the enzyme which induces the anti-inflammatory effect, production of PGE ₂ (prostaglandin 2, prostaglandin E2) was induced by using mouse- The expression levels of iNOS (inducible nitric oxide synthase), which induces the production of COX-2 (cyclooxygenase 2) and NO (nitric oxide), are measured by cytosol, nuclear and cytoplasm fractions (Western blot analysis).

Specifically, mouse-derived macrophages were cultured in DMEM containing 10% heat-inactivated FBS (fetal bovine serum) and penicillin G (100 IU / ml, Gibco co, USA) and streptomycin (100 μg / ml, Gibco co, USA) And the cells were cultured at 37 占 폚 for 24 hours using a 5% CO ₂ incubator (Sanyo, MCO175). The cultured broth ^{(1 × 10 6 cells / ml} ) to 1 ml each was separated in a 96-well plate, pearl Chellah Min G concentration by 5, 10, 20, and 40 μM each treatment, add 5% CO ₂ incubator in Lt; / RTI &gt; for 12 hours. To induce inflammatory responses after incubation, LPS was treated with 1 μg / ml and further incubated for 18 hours. The cultured macrophages were harvested and centrifuged at 2,000 rpm and 4 ° C for 2 minutes. After centrifugation, the supernatant was removed and the remaining pellet was resuspended in a PER-Mammalian Protein Extraction Buffer (PBS) containing protease inhibitor cocktail I (EMD Boisciences, USA) and 1 mM PMSF (phenylmethylsulfonyl fluoride) Extraction buffer, Pierce Biotechnology, USA) was added and homogenized. PMSF (0.1 mM PMSF, 5 mg / ml aprotinin, 5 mg / ml pepstatin A and 1 mg / ml PMSF) was added to 20 mM TrisHCl buffer (pH 7.4) containing the protein degradation inhibitor and PMSF. / ml chymostatin). The cytosolic fraction was then prepared by centrifuging the homogenized cell liquor at 15,000 x g and 4 ° C for 10 minutes.

In addition, NE-PER nuclear and cytoplasmic extraction reagents (Pierce Biotechnology, USA) were used for the production of nuclear and cytoplasmic fractions. LPS was cultured and cultured for 18 hours × 10 ⁶ cells / ml) were separated in a 60 mm dish, and the cells were collected and washed with PBS (phosphate-buffered saline). After washing, the cells were washed with RIPA buffer (RIPA buffer, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, 50 Mm TrisHCl [pH 7.4], 50 mM glycerophosphate, 20 mM NaF, Cells were lysed by treatment with 20 mM ethylene glycol tetraacetic acid (EGTA), 1 mM dithiothreitol [DTT], 1 mM Na ₃ VO ₄ , and protease inhibitors) and stirring at 4 ° C for 15 minutes. Thereafter, the mixture was centrifuged at 15,000 x g and 4 ° C for 15 minutes, and then, a supernatant was obtained.

The prepared cytoplasmic fractions, and nuclear and cytoplasmic fractions were stored at -70 ° C until used in the experiment.

<7-2> Confirmation of the change in the expression of the anti-inflammatory enzyme

Western blot analysis to determine the expression of iNOS and COX-2 from the cytoplasmic fractions obtained by the method of Example <7-1> was performed using PER-Mammalian protein extraction buffer containing proteolysis inhibitor and PMSF The protein content in the cytoplasmic fraction was quantitated using a Lowry protein assay kit (P5626, Sigma Chemical Co., USA). The protein solution separated in the same amount as above was separated by 12% SDS-PAGE and transferred to an ECL nitrocellulose membrane (Bio-Rad, USA). The ECL nitrocellulose membrane with the proteins was blocked with 5% skimmed milk and then washed. The antibody reaction was performed by adding primary antibody (COX-2 and iNOS) to the cells (SantaCruz Biotechnology, USA) After reacting at 4 ° C for 24 hours, the membrane was washed, and then a secondary antibody (HRP conjugated anti-rabbit antibody) was added to the washed membrane. After reacting for 1 hour And the signal of the antibody was detected using an ECL detection system (Amersham Pharmacia Biotech, USA) according to the protocol provided.

As a result, as shown in Fig. 4, the expression of β-actin, which is not related to inflammation, was not changed in all experimental groups. However, in case of iNOS and COX-2 protein expression related to inflammation, The expression level was changed. In other words, iNOS and COX-2 proteins were not expressed in the control group without LPS addition, but the expression of iNOS and COX-2 protein was rapidly increased when LPS was added. In addition, when the penicillinolide A of the present invention was added, it was confirmed that iNOS and COX-2 protein expression was decreased in a concentration-dependent manner. In comparison with the control group treated with LPS alone, the test group treated with 20 μM iNOS , And COX-2 was significantly decreased from the experimental group treated with 20 [mu] M (Fig. 4).

From the above results, it can be seen that the addition of penicillinolide A of the present invention reduces the expression amount of iNOS and COX-2 protein and consequently inhibits the expression of iNOS and COX-2 and inhibits inflammatory cytokines, .

< Example  8> Penicillinoide  Identification of the production and activation inhibition of inflammation-related proteins of A

The following experiment was conducted to confirm the inhibition of phosphorylation and degradation of IκB-α, a substance that inhibits nuclear translocation of NF-κB (p65), which is known to inhibit the expression of inflammatory proteins.

Specifically, the cytoplasmic fraction and the nuclear and cytoplasmic fractions were collected in the same manner as in Example <7-1>, and IκB-α and phosphorylated IκB-α (phosphorylated IκB-α, p-IκB- And the content of p65 in nuclear and cytoplasmic fractions were measured by Western blot analysis using antibodies against each.

In addition, the binding activity of NF-κB to DNA in the collected nuclear and cytoplasmic fractions was measured using a TransAM kit (TransAM kit, Active Motif, USA) according to the manufacturer's protocol Respectively.

As a result, as shown in FIG. 5A, it was confirmed that the expression of IκB-α was significantly reduced in the LPS-treated group compared to the control group, and the induction of inflammation by LPS reduced the decrease of IκB-α, . When penicilinoid A was treated, the expression of IκB-α was increased in a concentration-dependent manner, and the treatment of penicillinolide A inhibited the degradation of IκB-α induced by LPS . In addition, it was confirmed that the expression of p-IκB-α was increased in the treatment group treated with LPS compared with the group treated with no treatment, and the induction of inflammation by LPS increased the increase of p-IκB-α, That is, it was confirmed to induce activation. On the other hand, in the case of treatment with penicillinolide A, it was confirmed that the expression of p-IκB-a was decreased in a concentration-dependent manner, and the activation of IκB-a induced by LPS was inhibited by treatment with penicillinolide A (Fig. 5A).

In addition, as shown in FIG. 5B, it was confirmed that the expression of p65, that is, NF-κB was increased in the nuclear fraction of the group treated with LPS compared with the group treated with no treatment, , Inducing NF-kB translocation into the nucleus. On the other hand, in the case of treatment with penicillinolide A, the expression of p65 decreased in a concentration-dependent manner and it was confirmed that the migration of NF-κB into the nucleus induced by LPS by the treatment with penicillinolide A was inhibited (Fig. 5B).

In addition, as shown in FIG. 5C, the DNA binding activity of NF-κB was about 4-fold increased in the LPS-treated group compared to the untreated group, whereas the penicillinolide A treatment In one case, the degree of DNA binding of NF-κB was decreased in a dose-dependent manner, and the degree of DNA binding was significantly decreased from the experimental group treated with 10 μM to about 50% in the experimental group treated with 10 μM (FIG. 5C ).

From these results, it can be seen that penicilinoid A inhibits the degradation of IκB-α in cells, inhibits the production of p-IκB-α, ie, activation of IκB-α, and inhibits the migration of NF- And the DNA binding activity of NF-κB was weakened, and it was confirmed that the anti-inflammatory effect, that is, the anti-inflammatory effect, was excellent.

< Example  9> Penicillinoide  Identification of the production and activation inhibition of inflammation-related enzymes of A

The following experiments were performed to determine the expression and activity of hematoxylin-1 (Heme Oxygenase-1, HO-1), an inflammation-related enzyme.

Specifically, the cytoplasmic fraction, nuclear and cytoplasmic fractions were collected in the same manner as in Example <7-1>, and the mRNA content, HO-1 production amount, and HO-1 activity of HO- When phenylaciliyolide A was treated with 40 μM, the expression level of HO-1 was measured with time. As a positive control, cobalt protophorphyrin (CoPP), which promotes the expression of HO-1, was treated with 20 μM. Assay for HO activity of HO-1 was determined by adding NADPH and a rat liver cytosol and hemin as a biliverdin reductase to the microsomes obtained from the collected cells. Was added to the reaction mixture. The reaction was carried out under the dark condition and the temperature condition of 37 ° C for 1 hour and terminated by treating with 1 ml of chloroform. The extracted bilirubin values were determined by measuring the absorbance at 464 nm and 530 nm.

As a result, as shown in Fig. 6A, it was confirmed that when the positive control group CoPP was treated, the activity of HO-1 was remarkably increased as compared with the control group in which no treatment was carried out. When the treatment with penicilinoid A was performed, The activity of HO-1 was increased in a concentration-dependent manner, and 40 [mu] M was significantly higher than that of CoPP (Fig. 6A).

With respect to the production of HO-1, Western blot analysis for HO-1 was carried out in the same manner as in Example <7-2>, and the concentration of HO-1 Protein production was confirmed.

As a result, as shown in FIG. 6B, the amount of HO-1 produced increased in a concentration-dependent manner when penicillinolide A was treated, and when 40 μM was treated, as in the case of treatment with CoPP as a positive control, (Fig. 6B).

In addition, in the case of treatment with 40 μM, which was confirmed to have the greatest amount of production, the production amount of HO-1 according to the treatment time was examined. As shown in FIG. 6C, it was found that 6 hours after the treatment with penicilinoid A It was confirmed that HO-1 was produced from the next time, and a clearly increased band was observed at the lapse of 18 hours, confirming that the amount of HO-1 produced increased with the lapse of time according to the treatment with penicilanolide A (Fig. 6C).

As can be seen from the above, according to the present invention, Penicillium The novel penicillinolide A compound of the present invention can be mass-produced using the SF-5292 strain and can be used to develop an anti-inflammatory composition using the penicillinolide A compound of the present invention. have.

Korea Biotechnology Research Institute KCTC18284P 20140425

<110> Korea Research Institute of Bioscience and Biotechnology <120> Composition for the prevention and treatment of inflammatory          diseases comprising the penicillinolidea isolated from marine          fungi <130> 2014P-04-069 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 915 <212> RNA <213> Penicillium sp. SF-5292 <400> 1 ttttttctcc tccccccccc cccgaaatat cataagcgga ggaaaagaaa ccaacaggga 60 ttgccccagt aacggcgagt gaagcggcaa gagctcaaat ttgaaagctg gctccttcgg 120 ggtccgcatt gtaatttgta gaggatgctt cgggagcggt ccccatctaa gtgccctgga 180 acgggacgtc atagagggtg agaatcccgt atgggatggg gtgtccgcgc ccgtgtgaag 240 ctccttcgac gagtcgagtt gtttgggaat gcagctctaa atgggtggta aatttcatct 300 aaagctaaat attggccgga gaccgatagc gcacaagtag agtgatcgaa agatgaaaag 360 cactttgaaa agagagttaa aaagcacgtg aaattgttga aagggaagcg cttgcgacca 420 gactcgctcg cggggttcag ccggcattcg tgccggtgta tttccccgcg ggcgggccag 480 cgtcggtttg ggcggtcggt caaaggccct cggaaggtaa cgcccctagg ggcgtcttat 540 agccgagggt gcaatgcgac ctgcctagac cgaggaacgc gcttcggctc ggacgctggc 600 ataatggtcg taagcgaccc gtcttgaaac acggaccaag gagtctaaca tctacgcgag 660 tgttcgggtg tcaaacccgt gcgcgaagtg aaagcgaacg gaggtgggaa cccttacggg 720 tgcaccatcg accgatcctg aagtcttcgg atggatttga gtaagagcgt agctgttggg 780 acccgaaaga tggtgaacta tgcctgaata gggcgaagcc agaggaaact ctggtggagg 840 ctcgtagcgg ttctgacgtg caaatcgatc gtcgaatttg ggtatagggg cgaaagacta 900 atcgaaccat ctagc 915

Claims (5)

A penicillinolide A compound represented by the following formula (2) or a pharmaceutically acceptable salt thereof:
(2)

.
The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is isolated from a strain of Penicillium genus SF-5292 deposited with Accession No. KCTC 18284P.
1) culturing the genus Penicillium genus SF-5292;
2) extracting the culture obtained in step 1) with methylethylketone; And
3) Separation of the penicillinolide A compound represented by the following formula 2, which comprises separating the methyl ethyl ketone extract obtained in the step 2) by column chromatography:
(2)

.
A medicament for the prophylaxis and treatment of dermatitis, allergies, systemic lupus erythematosus, retinitis, gastritis, hepatitis, enteritis, pancreatitis, nephritis and combinations thereof, which comprises the compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient Gt;
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