KR101498671B1 - Novel compound having antimicrobial activity - Google Patents

Abstract

The present invention relates to a novel compound exhibiting an antibacterial activity or a pharmaceutically acceptable salt thereof, a Penicillium sp. Strain extract containing the compound, a fraction of the extract, the compound or a pharmaceutically acceptable salt thereof as an active ingredient A pharmaceutical composition for treating or preventing pyruvic infections, a composition for inhibiting the activity of Fab-I activity of an enoyl-ACP reductase, and a method for producing the compound. Since the compounds of the present invention inhibit the activity of the enoyl-ACP reductase and exhibit an antimicrobial activity that inhibits the growth of Bacillus and Staphylococcus aureus causing food poisoning or purulent infectious diseases, the compounds are useful for the development of a therapeutic agent for food poisoning and purulent infectious diseases It can be widely used.

Description

Novel compound having antimicrobial activity < RTI ID = 0.0 >

The present invention relates to an antimicrobial compound, and more specifically, the present invention relates to a novel compound exhibiting an antibacterial activity or a pharmaceutically acceptable salt thereof, a Penicillium genus extract containing the compound, a fraction of the extract, Or a pharmaceutically acceptable salt thereof as an active ingredient, a pharmaceutical composition for the treatment of food poisoning, a pharmaceutical composition for the prevention or treatment of a pyogenic infectious disease, a composition for inhibiting Fabo activity of an enoyl-ACP reductase, .

As the use of antibiotics is becoming more frequent, the rate of resistance development has also increased. Thus, finding and developing new antibiotics is becoming an important issue in order to cope with the problems that threaten our essential life.

In order to develop new antibiotics, developing antibiotics, rather than creating derivatives through modification of existing antibiotics, is a good approach to overcome resistant bacteria. Among these potential bacterial targets, bacterial fatty acid biosynthesis has attracted attention as a new antibiotic target (Heath, R. J., et al., Prog. Lipid Res., 2001, 40: 467-497).

Since fatty acid biosynthesis is an important part of cell membrane formation, it plays an indispensable role in bacterial growth. And the process of biosynthesis of these fatty acids in cells is an essential biochemical process in all living cells. The fatty acid biosynthesis process in higher animals and bacteria has a slight difference in enzyme relatedness. In higher animals, fatty acids are synthesized by a single large polypeptide called the fatty acid enzyme (FAS), but in the bacterial system several enzymes with one function are present separately and involved in each reaction. These differences lead to the selection between the host mammal and the bacteria in the fatty acid biosynthetic process (Campbell, J. W., et al., Annu. Rev. Microbiol., 2001, 55: 305-332).

Fatty acid biosynthesis consists of four steps: condensation, reduction, dehydration and reduction. As a final step in the fatty acid biosynthesis, trans-2-enoyl-ACP is converted to acyl-ACP by an enyl-ACP reductase FabI, which is a kind of NADH (NADPH) -dependent enyl- The process of reduction occurs. The trans-2-enoyl-ACP (FabI) enzyme used in the last step is the major regulatory point in the overall FAS synthesis step as a rate determining step (Heath, RJ, et al., J. Biol. Chem , 1995, 270: 26538-26542).

Bacterial enoyyl-ACP reductase, which promotes the final and rate-determining step in bacterial fatty acid synthesis, has been demonstrated as a novel target in the development of antimicrobial agents. The antimicrobial target of isoniazid, which has been used for the treatment of triclosan, a broad-spectrum biocide for many consumer products and 50 years of tuberculosis, has been shown to be an enoyl-ACP reductase. The Enyl-ACP reductase has four types of isoforms: Fab I, FabK, FabL, and FabV. Among them, FabI is well conserved among important pathogenic bacteria including methicillin-resistant Staphylococcus aureus (MRSA), and various FabI synthesis inhibitors have been reported to have broad spectrum antibacterial activity.

Under these circumstances, the present inventors have made intensive research to develop a novel antimicrobial compound capable of inhibiting the proliferation of harmful bacteria by effectively inhibiting the activity of the enyl-ACP reductase. As a result, it has been found that Penicillium verruculosum F375 strain of the present invention can effectively inhibit the activity of the enoyl-ACP reductase and inhibit the growth of Staphylococcus aureus and Bacillus. The present invention has been completed based on this finding.

It is an object of the present invention to provide a novel compound exhibiting antibacterial activity or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide an antimicrobial composition comprising a Penicillium sp. Strain extract containing the compound, a fraction of the extract, the compound or a pharmaceutically acceptable salt thereof as an active ingredient.

It is still another object of the present invention to provide a pharmaceutical composition for treating food poisoning, comprising the extract of Penicillium sp. Containing the above compound, the fraction of the extract, the compound or a pharmaceutically acceptable salt thereof as an active ingredient.

Yet another object of the present invention is to provide a pharmaceutical composition for preventing or treating a pyogenic infectious disease comprising an extract of a genus of the genus Penicillium sp. Containing the above compound, a fraction of the extract, the compound or a pharmaceutically acceptable salt thereof as an active ingredient .

Yet another object of the present invention is to provide a composition for inhibiting Fabo activity of an enyl-ACP reductase comprising a Penicillium sp. Strain extract, a fraction of the above extract, the compound or a pharmaceutically acceptable salt thereof .

It is a further object of the present invention to provide a process for the preparation of said compounds.

In one embodiment for achieving the above object, the present invention provides a compound of the following general formula (I) or a pharmaceutically acceptable salt thereof.

(I)

(I)

In this formula,

이다.
R is

to be.

More specifically, the compound provided by the present invention may be a compound represented by the following general formula (II) or (III).

(II)

(II)

(III)

(III)

The present inventors extracted Penicillium verruculosum F375 strain, a kind of Penicillium verruculosum F375 strain, by extracting with acetone, fractionating it with ethyl acetate, and then using Sephadex LH-20 column chromatography and silica gel 60 RP-18 F254 plate , Two compounds capable of inhibiting the activity of the enyl-ACP reductase were purified and identified. As a result, it was found that one of the compounds was 2,3,7-tetrahydronaphthalene represented by the formula (II) Trihydroxy-9-methoxy-6H-dibenzo [b, d] pyran-6-one) (Hereinafter referred to as " Verrulactone A ") at C-1 of the formula (III) and the other is 2,3,7-trihydroxy-9 -Methoxy-6H-dibenzo [b, d] pyran-6-one of 2,3,7-trihydroxy-9-methoxy- (Hereinafter referred to as "Verulaton B ctone B) ").

The above-identified berulactones A and B are each a symmetric or asymmetric dimer of the novel ether or alternarol. Altera riol was isolated from fungal endophyte, Alternaria sp. Several derivatives of the alkanediol have been reported from fungi, lichens and plants. (9-O-methylalternauriol) derived from plants Anthocleista djalonensis and Penicillium diversum, 2-chloro-2-methyl-2-butanol derived from the antifungal Lachnum palmae, Graphislactone F and E from 2-chloro-9-O-methylalternariol, and the lichen, Graphis prunicola, have been reported. The dimeric compound of the ether allyol was first identified in the present invention. Altera ranol, etheraliol 9-sulfate and 9-O-methyletheralanil are cytotoxic in cancer cells and have been reported to inhibit protein kinase. Altera riol also inhibited the cell cycle and not only showed an inhibitory effect on cell proliferation, but also had an estrogenic potential. However, the antimicrobial activity of the ether and the derivative thereof has not yet been reported.

The term " Penicillium sp. Strain " of the present invention means a microorganism belonging to a fungus belonging to a blue mold having a broom-shaped conidial bag. The Pericillium sp. Strain may preferably be Penicillium verruculosum, more preferably Penicillium verruculosum F375 strain, but is not particularly limited thereto. For the purpose of the present invention, the strain of the genus Penicillium can be used as a parent strain for producing the antimicrobial compound provided in the present invention, but is not particularly limited thereto.

The term "extract" of the present invention means that the target substance is dissolved in water, a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol or butanol), an organic solvent (hexane, acetone, chloroform, methyl acetate, And the like. The resultant product includes all of the extract, the diluted solution or concentrate of the extract, the dried product obtained by drying the extract, or the adjusted product or the purified product. For the purpose of the present invention, the extract is a result obtained by extracting a strain of Penicillium sp. With an organic solvent. The extract may include the antimicrobial compound provided by the present invention, but is not particularly limited thereto.

In another aspect, the present invention relates to an antimicrobial composition comprising an extract of a genus Penicillium containing Berrolactone A and B, a fraction of the extract, the compound or a pharmaceutically acceptable salt thereof as an active ingredient, There is provided a composition for inhibiting ACP reductase FabI activity.

Berlactones A and B derived from the Penicillium sp. Strain extract of the present invention not only exhibit the effect of inhibiting the activity of the enyl-ACP reductase FabI derived from Staphylococcus aureus (Example 3-1) (Example 3-2) as well as staphylococcus and methicillin-resistant Staphylococcus aureus, as well as cell growth of Bacillus cereus strain (Example 3-2).

The term "fraction " of the present invention means a product obtained by a fractionation method for separating a specific component or a specific group from a mixture containing various constituents. In the present invention, preferably, the extract of the genus Penicillium is fractionated by a solvent fractionation method using a solvent such as n-hexane or ethyl acetate, and includes both a polar solvent fraction and a non-polar solvent fraction. Specifically, Methanol fraction, ethyl acetate fraction and the like can all be used.

The term "Staphylococcus aureus " as used herein means a gram-positive anaerobic bacterium which forms a cell aggregate (lump) as it grows. The staphylococcus aureus is a bacterium belonging to the genus Escherichia, Generally, it produces exotoxins that are heat-resistant and cause food poisoning, and secrete toxins (luocosidine), hemolysin, coagulase, etc. which kill phagocytes and can cause a pneumonia infection from the resistance of infected host cells. Methicillin-resistant Staphylococcus aureus (MRSA), which is resistant to most antibiotics, also belongs to Staphylococcus aureus. For the purpose of the present invention, the Staphylococcus aureus can be used as a target of the antimicrobial compound provided in the present invention, But is not particularly limited thereto.

The term "Bacillus" in the present invention means a generic name of a group of bacteria produced in a bar. In general, about 148 species are known to live in various living environments such as people's living environment and soil. Currently, there are Bacillus subtilis and Bacillus cereus species. For the purpose of the present invention, the bacterium may be used as a target of the antimicrobial compound provided in the present invention, but is not particularly limited thereto.

The term "food poisoning" of the present invention means a series of syndromes caused by toxic substances contained in ingested food. Depending on the cause, bacterial food poisoning which causes infection by bacteria itself or toxins produced by bacteria, natural poisoning by animal or vegetable toxins in nature, chemical poisoning which causes symptoms by artificial chemicals Respectively. For purposes of the present invention, the food poisoning may be bacterial food poisoning caused by Staphylococcus aureus or Bacillus that is directly affected by the compounds of the present invention, but is not limited thereto.

The term " purulent infectious disease " in the present invention refers to a disease that causes pain when a bacterium (enterococcus) is infected in a traumatic area and pyuria (pus) occurs. Examples of the diseases belonging to the above-mentioned pyogenic infectious diseases include otitis media, cystitis, purulent pimples, acanthosis, rhizomes, roots, rhinitis, and rhinopathies. Examples of the causative agents are yellow staphylococci and pseudomonas aeruginosa, But is not particularly limited thereto.

The term "antibacterial" or "antibacterial activity" of the present invention means a property of resisting microorganisms such as bacteria or fungi. More specifically, antibiotic or the like means a property of inhibiting growth or proliferation of bacteria. For purposes of the present invention, the antimicrobial or antimicrobial activity is preferably a microorganism essentially comprising an enoyl-ACP reductase, more preferably a Staphylococcus aureus or Bacillus, most preferably a methicillin-resistant Staphylococcus aureus, May be used as a property to inhibit growth or proliferation of Staphylococcus aureus RN4220, bacillus cereus, etc., but is not particularly limited thereto.

In another aspect, the present invention provides a pharmaceutical composition for treating food poisoning, comprising a penicillium genus extract containing the compound, a fraction of the extract, the compound or a pharmaceutically acceptable salt thereof as an active ingredient, Or a pharmaceutical composition for therapeutic use.

As described above, the Berlacktons A and B provided by the present invention exhibit activity to inhibit cell growth of Bacillus cereus as well as Staphylococcus aureus and methicillin-resistant Staphylococcus aureus. Therefore, the Berlacktons A and B Can be used not only to treat the symptoms of food poisoning which can be caused by Staphylococcus aureus and Bacillus cereus, but also to treat or prevent the infectious diseases caused by Staphylococcus aureus. It is also obvious to those skilled in the art that the treatment or prophylactic effects of food poisoning and pyogenic infections can be manifested not only by Berlactones A and B but also by the Penicillium sp. Strain extract or fractions thereof .

In addition, the pharmaceutical compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions. The pharmaceutical composition of the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method have. In the present invention, the carrier, excipient and diluent which may be contained in the pharmaceutical composition 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. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, Sucrose, lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use may include various excipients such as wetting agents, sweetening agents, fragrances, preservatives, etc. in addition to water and liquid paraffin, which are simple diluents commonly used in suspension agents, solutions, emulsions and syrups have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

In addition, the pharmaceutical composition of the present invention can be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically) depending on the intended method, The severity of the disease, the form of the drug, the route of administration and the period of time, may be appropriately selected by those skilled in the art. If necessary, it may be administered once or several times a day, and may be used alone or in combination with methods using surgery, hormone therapy, drug therapy and biological response modifiers for the prevention or treatment of pathogenic bacteria and resistant bacteria . Preferably, the composition of the present invention may be administered at 0.1 to 50 mg / kg, more preferably 5 to 30 mg / kg. The most preferred dosage of the composition of the present invention is about 30 mg / kg. As described above, the administration route of the present invention is also not limited, but it can be preferably administered parenterally, and more preferably, it can be administered by a subcutaneous injection method.

In another aspect, the present invention relates to a composition for preventing or improving a purulent infectious disease comprising, as an active ingredient, a Penicillium sp. Strain extract containing the compound, a fraction of the extract, the compound or a pharmaceutically acceptable salt thereof .

The term "functional food " of the present invention is the same term as " food for special health use (FoSHU) ", and includes medical and medical effects It means high food. In the present invention, the term " health food "refers to a food having an active health promotion or promotion effect as compared to a general food, and a health supplement food refers to a food for health assistance. Accordingly, the terms of functional foods, health foods, and health supplement foods are used. The foods may be prepared in various forms such as tablets, capsules, powders, granules, liquids, have.

The extract of Pericillium sp. Strain, the fraction of the extract, the compound or the pharmaceutically acceptable salt of the compound comprising Berlactone A and B is contained in an amount of 0.01 to 100% by weight, Is contained in an amount of 1 to 80% by weight. When the food is a beverage, it may be contained in a proportion of 1 to 30 g, preferably 3 to 20 g based on 100 ml. In addition, the composition may contain additional ingredients which are commonly used in food compositions and which can improve odor, taste, vision and the like. For example, vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, panthotenic acid and the like. In addition, it may include minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu) It may also include amino acids such as lysine, tryptophan, cysteine, valine, and the like. In addition, it is also possible to use antiseptics (such as potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate), disinfectants (such as bleaching powder and highly bleached white powder, sodium hypochlorite), antioxidants (butylhydroxyanilide (BHA), butylhydroxytoluene BHT), etc.), coloring agents (such as tar pigments), coloring agents (such as sodium nitrite and sodium acetic acid), bleaching agents (sodium sulfite), seasoning (such as MSG sodium glutamate), sweeteners (such as hypoglycemia, , Food additives such as flavorings (vanillin, lactones, etc.), swelling agents (alum, potassium hydrogen D-tartrate), emulsifiers, emulsifiers, thickeners, encapsulating agents, gum bases, foam inhibitors, ) Can be added. The additives are selected according to the type of food and used in an appropriate amount.

A food composition comprising a Pericillium sp. Strain extract containing the compound of formula (I), a fraction thereof, a compound or a pharmaceutically acceptable salt of the compound may be used to treat various types of degenerative neurological diseases A functional food for prevention or improvement can be produced.

As a concrete example, the above-mentioned composition can be used to produce a processed food having good shelf-life while modifying the characteristics of agricultural products, livestock products or aquatic products. Such processed foods include, for example, confectionery, drinks, liquor, fermented foods, canned foods, processed milk products, processed marine foods, noodles and the like. The sweets include biscuits, pies, cakes, breads, candies, jellies, gums, cereals (including dinner utensils such as cereal flakes). Drinks include carbonated beverages, functional ionic beverages, juices (such as apples, pears, grapes, aloes, citrus fruits, peaches, carrots, tomato juices, etc.) and sikhye. The mainstream includes sake, whiskey, shochu, beer, liquor, and fruit wine. Fermented foods include soy sauce, miso, and kochujang. Canned products include canned products (eg, tuna, mackerel, saury, canned fish, etc.), canned products (canned beef, pork, chicken, turkey canned food) and canned products (corn, peach and canned pineapple). Milk processed foods include cheese, butter, yogurt and the like. Meat-processed foods include pork cutlet, beef cutlet, chicken cutlet, sausage. Sweet and sour pork, nuggets, and nubani. And noodles such as sealed packaging raw noodles. In addition, the composition may be used in retort food, soup and the like.

In another aspect, the present invention provides a process for preparing said compound. Specifically, a method for producing the compound of formula (I) comprises: (a) culturing a strain of the genus Penicillium sp. To obtain a culture; (b) extracting the culture with acetone to obtain an extract, and fractionating the extract with ethyl acetate to obtain a fraction; And (c) applying said fraction to chromatography to obtain the compound of formula (I).

In the above method, the Penicillium verruculosum F375 strain is preferably used, and the chromatography is performed using Sephadex LH-20 column chromatography and silica gel 60 RP-18 F254 plate It is preferable to apply the thin layer chromatography used sequentially.

In the present invention, the culture of Penicillium sp. Strain for producing the compound of the present invention can be cultured in a medium containing a nutrient source that can be used for conventional microorganisms. As a nutrient source of the strain, a nutrient source commonly used in the art can be used without limitation, and a known nutrient source conventionally used for the cultivation of a fungus can be used. Examples of the carbon source include glucose, syrup, dextrin, starch, molasses, animal oil, vegetable oil and the like. Examples of nitrogen sources include bran, soybean meal, wheat, malt, cottonseed oil, Ammonium, sodium nitrate, urea, and the like can be used. It is very effective to add sodium chloride, potassium, magnesium, cobalt, chlorine, phosphoric acid, sulfuric acid and other inorganic salts to promote ion production as needed. As the culture method, it is possible to cultivate shaking or stationary culture under aerobic conditions.

The culturing temperature may be slightly different depending on the conditions when culturing under the above conditions, but it is suitably cultured at 20 to 37 ° C, preferably at 26 to 30 ° C. The incubation period can also be cultured for a known period of time as used in the art, and the period can be adjusted as necessary. Preferably, in the case of shaking culture or stationary culture, it is possible to cultivate for 4 days to 7 days, and it is confirmed that the yield of the compound of the present invention reaches the maximum value at the above culture period.

From the preferred embodiment of the present invention, the compound of the present invention can be obtained by culturing the strain under the conditions described above (Example 1), and the compound of the present invention can exist not only in the culture medium but also in the cell Therefore, it can be efficiently separated and purified by the following method for effective extraction.

The method for extracting and purifying a compound can be used without any limitation in a method commonly used in the art, and it is possible to control the yield and the yield by changing the kind of medium, culture condition, extraction and purification method, Do. In a preferred embodiment of the present invention, extraction is carried out using acetone and ethyl acetate. Specific methods are as follows.

After the organic solvent such as acetone was added to the culture solution and the mycelial body and stirred to extract an active ingredient from the cells, the acetone was evaporated and solvent extraction was performed with ethyl acetate, and then the ethyl acetate solvent layer containing the active ingredient was concentrated under reduced pressure After removal of the ethyl acetate, the material of the present invention was obtained by sequential application to thin layer chromatography using Sephadex LH-20 column chromatography and silica gel 60 RP-18 F254 plate (Example 1). The inventors of the present invention confirmed that it is possible to effectively extract and purify Berlactone A and B from Penicillium verruculosum F375 strain using this method.

Since the compounds of the present invention inhibit the activity of the enoyl-ACP reductase and exhibit an antimicrobial activity that inhibits the growth of Bacillus and Staphylococcus aureus causing food poisoning or purulent infectious diseases, the compounds are useful for the development of a therapeutic agent for food poisoning and purulent infectious diseases It can be widely used.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic diagram showing the HMBC and NOE associations of compounds 1 and 2. FIG.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for describing the present invention more specifically, and the scope of the present invention is not limited by these examples.

Example  One: Penicillium verruculosum  Isolation of F375 strain-derived compounds

Penicillium verruculosum F375 strain was isolated from the soil collected from the Kingang mine, Yangdong-myeon, Yangpyeong-gun, Gyeonggi-do, Korea and the isolated strain was cultivated in 80 mL culture medium (glucose 2%, polypeptone 0.5%, yeast extract 0.2%, KH ₂ PO ₄ 0.1% ₄ · 7H ₂ O 0.05%, pH 5.6-5.8) and cultured at 28 ° C for 3 days with shaking. 5 mL of the cultured culture was again inoculated into a 100 mL culture medium, and then cultured for 14 days under the same conditions. As a result, berulactones A and B were produced after 10 days. After the completion of the culture, acetone was added to the culture so that the final concentration became 50%, and the mixture was centrifuged at 8000 rpm for 20 minutes to obtain a supernatant. The supernatant was concentrated in vacuo and the concentrate was mixed with an equal volume of ethyl acetate to give an ethyl acetate fraction. The procedure was repeated three times. The ethyl acetate fraction obtained was dried in vacuo to give 2.8 g of the dried product. The dried product was applied to Sephadex LH-20 (Amersham Biosciences) column chromatography and eluted with methanol to obtain an active fraction. The resulting active fraction was concentrated in vacuo to give a yellow residue (560 mg). The residue was applied again to Sephadex LH-20 column chromatography and eluted with methanol to give two active fractions. Each of the active fractions obtained was concentrated in vacuo and thin layer chromatography (TLC) was performed using silica gel 60 RP-18 F254 plate (Merck No. 1.15389.0001, Darmstadt, Germany). At this time, ACN-water (55:45) containing 0.1% TFA was used as a developing solvent. As a result, compound 1 (6 mg) and compound 2 (10 mg) were obtained in the form of yellow powder, respectively.

The obtained compounds 1 and 2 were applied to an analytical HPLC column (4.6 x 250 mm, S-4 uM, YMC C18) chromatography, in which ACN-water (45: 55) was used, and the flow rate was set at 0.9 ml / min. As a result of the analysis, Compounds 1 and 2 exhibited a retention time of 10.1 minutes and 11.9 minutes, respectively, and a purity of 99.5% or more at 210 nm.

Example  2: Spectroscopic analysis of compounds 1 and 2

Compounds 1 and 2 obtained in Example 1 were analyzed spectroscopically using ¹ H and ¹³ C NMR and high resolution ESI-MS [(MH) ^- , 545.0726 m / z (+0.1 mmu error) (Table 1).

Table 1 shows the ¹ H (500MHz) and ¹³ C (125MHz) NMR spectral data of compounds 1 and 2, under DMSO-d6.

First, from the results of the above Table 1, it was confirmed that Compound 1 represented the molecular formula of C ₂₈ H ₁₈ O ₁₂ , and IR analysis revealed that Compound 1 contained a carbonyl group (1662 / cm) and a hydroxyl group (3448 / As shown in Fig.

¹ H and ¹³ C NMR, ¹ H- ¹ H COZY, DEPT, and HMQC data show that there are two meta-position-coupled protons, which are 1, 2, 3, 5-disubstituted benzene rings (1, 2,3,5-tetrasubstituted benzene ring) [delta] 6.38, d, J = 2.1; [delta] 100.1 and [delta] 6.41, d, J = 2.1; δ 100.0], the isolated room stars meth (δ 6.98, s; δ 103.2 ), methoxy (δ 3.38, s; δ 55.0 ) , and possibly one exchange hydrogen (δ 11.81, s), 9 of sp ² 4 Tea (Δ 165.9, 163.8, 148.7, 145.4, 141.4, 137.8, 121.3, 107.8, and 98.7) and one carbonyl carbon (δ 164.7). The connection of these partial structures and atoms was determined by HMBC spectral data analysis (Fig. 1). BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic diagram showing the HMBC and NOE associations of compounds 1 and 2. FIG.

Of the two meta-coupled protons, the protons showing δ 6.41 (H-8) were δ 165.9 (C-9), 163.8 (C-9) as well as another meta-coupling methane carbon showing δ 100.1 (C-7) and three sp ² represents a 98.7 (C-6a) and a quaternary carbon (sp ² quaternary carbon) and long-distance mating. Exchangeable hydrogens showing δ 11.81 (7-OH) showed Heteronuclear multiple bond correlation (HMBC) associations with C-6a, C-7 and C-8. This suggests the presence of exchangeable hydrogen at the ortho position of H-8. Another meta-coupling proton showing δ 6.38 (H-10) was a long-distance pair with C-9, which is associated with C-6a, C-8 and methoxy protons (9-OCH3). This indicated that the methoxy group was located between H-8 and H-10. The position of the methoxy group was confirmed by nuclear overhauser effect (NOE) between the methoxy proton and H-8 and H-10.

The hydrogen-bonded hydroxy proton (δ 11.81) of 7-OH along with the ¹³ C-NMR chemical shift (δ 98.7) of C-6a suggested that the carbonyl group (C-6) was attached to C-6a. On the other hand, the isolated aromatics with δ 6.98 (H-4) show three oxidized sp ² -dispersed carbons with δ 148.7 (C-3), δ 145.4 (C-2) and δ 141.4 -10 and showed δ 107.8 (C-10b) which are not oxidized showing a quaternary sp ² carbon and a strong HMBC correlation to the long-range coupling. These spectral data showed the connection between C-10a and C-10b. H-4 also showed weak HMBC associations with two sp ² quadrants showing δ 137.8 (C-10a) and 121.3 (C-1). Chemical shifts (DE 121.3) of DEPT and C-1 indicated that C-1 was not both hydrogenated and hydroxylated. These spectral data suggest that the C-1 is a symmetric dimer. In fact, ion peaks showing m / z 545 [MH] ^- , 547 [M + H] ⁺ and 569 [M + Na] ⁺ were observed in ESI- M + H] < ⁺ > was also detected.

Accordingly, the compound 1 is a 2,3,7-trihydroxy-9-methoxy-6H-dibenzo [b, d] pyran-6-one (2,3,7-trihydroxy -9-methoxy-6H-dibenzo [b, d] pyran-6-one) at C-1 and named it "Verrulactone A".

Next, the molecular formula of Compound 2 was identified in the same manner, and IR analysis was conducted. As a result, Compound 2 was confirmed to contain a carbonyl group (1666 / cm) and a hydroxyl group (3444 / cm). ^{The 1} H and ¹³ C NMR data consisted of two sets of signals, confirming a different set of NMR signals similar to those of compound 1 except for C-1. The above ¹ H and ¹³ C NMR, ¹ H- ¹ H COZY, DEPT and HMQC data showed the presence of four meta-coupled protons, indicating the presence of two 1, 2, 3, 5-substituted benzene rings 1, 2, 3, 5-tetrasubstituted benzene ring) [delta] 6.38, d, J = 2.3; [delta] 6.41, d, J = 2.3 and [delta] 6.57, d, J = 2.1; ? 7.08, d, J = 2.1], two isolated aromatic methines (? 6.95, s;? 103.2 and? 7.66, s;? 108.2), two methoxy , s; δ 54.9 and δ 3.95 s; δ 56.0), two exchangeable protons (δ 11.87, s and δ 11.38, s), 16 sp ² quadratic carbons (δ 166.8, 165.9, 163.8 , 163.7, 148.3, 147.0, 144.9, 143.6, 142.2, 142.1, 138.0, 137.4, 116.5, 113.5, 109.2, 108.3, 98.8, and 98.4) and two carbonyl carbon (? 164.8 and 164.7) . The presence of 1 monomeric unit was confirmed from HMBC spectroscopic data. The remaining 1H and < 13 > C NMR data were similar to the data of 1 unit of unit.

On the other hand, the difference between compounds 1 and 2 is one of the meta-coupling protons showing δ 7.08 (H-5 '), δ 3.95 (6'-OCH ₃ ), another isolated aromatic methane showing δ 7.66 ) Were moved to a low field. The connection of the remaining partial structures and elements was determined by HMBC spectroscopic data (Table 1 and Figure 1). The HMBC data of the 6'-OCH ₃ exchangeable protons showing two meta-coupling protons (H-5 'and H-7'), δ 11.38 (8'-OH) 3, 5-tetrasubstituted benzene ring in the presence of 3, 5-tetrasubstituted benzene rings. However, H-4 'has not only three oxidized sp ² quaternary carbons showing δ 147.0 (C-10'a), δ 143.6 (C-3') and δ 142.2 in -4'b) the visible quaternary sp ² carbon in the HMBC and showed a strong relationship, quaternary sp ² carbon exhibit δ 109.2 (C-4'a) showed no otherwise. These data show that C-1 and C-4 'are not connected but C-1 and C-1' are connected to induce the formation of an asymmetric dimer. These structures were confirmed as a nuclear overhaul effect (NOE) between H-4 ', H-5' and 6-OCH ₃ .

Accordingly, the compound 2 is a 2,3,7-trihydroxy-9-methoxy-6H-dibenzo [b, d] pyran-6-one represented by the general formula (III) 9-methoxy-6H-dibenzo [b, d] pyran-6-one) was named as "Verrulactone B".

Example  3: Berulakton  Antibacterial activity of A and B

Example  3-1: Derived from Staphylococcus aureus Enoil - ACP  Reducing enzyme Fabi Inhibitory activity against

The inhibitory activity of berulactones A and B against Staphylococcus aureus-based mono-ACP reductase Fab I was measured

Specifically, 200 μM to-NAC thioester and 200 μM NADPH in 50 mM sodium acetate buffer (pH 6.5) were added to 150 nM of the enoyl-ACP reductase Fab I from S. aureus, the enoyl-ACP reduction from S. pneumoniae Enzyme FabK or Enoyl-ACP reductase InhA derived from M. tuberculosis was added to a 96-well microplate, and Berulactone A or B dissolved in DMSO was added to each well and allowed to react. Then, The rate of increase in the amount of NADPH was measured at 340 nm using a microtiter ELISA reader using SOFTmax PRO software (Molecular Devices, Calif., USA) at 30 ° C, Were substituted into the following formula to calculate the inhibitory activity and to compare them (Table 2).

Inhibitory activity (%) = 100 x [1 - (rate in the presence of compound / rate in untreated control)]

Inhibitory Activity of Berulactones A and B on Enyl-ACP Reducing Enzymes Enoyl-ACP reductase IC ₅₀ ([mu] M) Fabi FabK InhA Berulactone A
Berulactone B 0.92
1.41 12.52
5.02 8.64
6.31

As shown in Table 2, for the enoyl-ACP reductase Fab I, berulactone A exhibited an IC ₅₀ value of 0.92 μM, Berulactone B exhibited an IC ₅₀ value of 1.14 μM, and concentration- Lt; RTI ID = 0.0 > Fab-1. ≪ / RTI >

Example  3-2: Cell growth inhibitory activity of Staphylococcus aureus and methicillin-resistant Staphylococcus aureus

It was confirmed that berulactones A and B can inhibit cell growth of various bacteria including Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA) (Table 3).

Cell growth inhibitory activity of berulactones A and B against various bacteria Target bacteria
MIC ([mu] g / ml) Berulactone A Berulactone B S. aureus RN 4220
S. aureus 209
MRSA CCARM 3167
MRSA CCARM 3506
MRSA CCARM 3505
Bacillus subtilis KCTC 1021
Bacillus subtilis KCTC 1661
S. pneumoniae KCTC 5412 8
16
16
16
32
32
8
64 16
32
32
32
8
16
16
64

As shown in Table 3 above, Berulactone A exhibited a minimal inhibitory concentration (MIC) of 8-16 μg / mL and was resistant to Staphylococcus aureus RN4220 containing methicillin-resistant Staphylococcus aureus (MRSA) CCARM 3167 Showed antibacterial activity against bacillus cereus KCTC 1661 but failed to inhibit the growth of E. coli KCTC 1683 and Pseudomonas aeruginosa KCTC 2004.

In addition, berulactone B exhibited a MIC of 16-32 μg / mL and showed about twice as much antibacterial activity as berlactone A against Staphylococcus aureus and methicillin-resistant Staphylococcus aureus.

The results showed a reliable association between Fab ' s inhibition and antibacterial activity of Berulactones A and B.

Claims (15)

A compound of the following general formula (II) or (III): or a pharmaceutically acceptable salt thereof:
In general formula (II)

In the formula (III)

The method according to claim 1,
Or an acyl-carrier protein (ACP) reductase, or a pharmaceutically acceptable salt thereof.
3. The method of claim 2,
Wherein said enol-ACP reductase is an enoyl-ACP reductase FabI, an enoyl-ACP reductase FabK, an enoyl-ACP reductase FabL or an enoyl-ACP reductase FabV, or a pharmaceutically acceptable salt thereof salt.
An antimicrobial composition comprising an extract of a strain of Penicillium sp. Containing the compound of claim 1, a fraction of the extract, the compound or a pharmaceutically acceptable salt of the compound as an active ingredient.
5. The method of claim 4,
Wherein the composition exhibits an antimicrobial activity against Staphylococcus aureus or Bacillus.
6. The method of claim 5,
Wherein said Staphylococcus aureus is methicillin resistant Staphylococcus aureus or Staphylococcus aureus RN4220.
6. The method of claim 5,
Wherein the Bacillus bacterium is bacillus cereus.
A pharmaceutical composition for the treatment of food poisoning comprising an extract of a strain of Penicillium sp. Containing the compound of claim 1, a fraction of the extract, the compound or a pharmaceutically acceptable salt of the compound as an active ingredient.
A pharmaceutical composition for treating a pyogenic infectious disease, comprising an extract of a strain of Penicillium sp. Containing the compound of claim 1, a fraction of the extract, the compound or a pharmaceutically acceptable salt of the compound as an active ingredient.
10. The method of claim 9,
Wherein the pyogenic infectious disease is selected from the group consisting of otitis media, cystitis, purulent acne, ache, spikelet, rods and rhinitis, rash and lymphadenitis.
10. The method according to claim 8 or 9,
Lt; RTI ID = 0.0 > pharmaceutically < / RTI > acceptable carrier.
In vitro , comprising an extract of a strain of Penicillium sp. Containing the compound of claim 1, a fraction of said extract, said compound or a pharmaceutically acceptable salt of said compound, A composition for inhibiting a reducing enzyme FabI activity.
(a) culturing a strain of Penicillium sp, to obtain a culture;
(b) extracting the culture with acetone to obtain an extract, and fractionating the extract with ethyl acetate to obtain a fraction; And
(c) applying the fractions to chromatography to obtain a compound of claim 1.
14. The method of claim 13,
Wherein said Penicillium spp. Strain is Penicillium verruculosum F375.
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