KR101792239B1 - Antibacterial agent comprising 7,10-epoxy-octadeca-7,9-dienoic acid and antibiotics - Google Patents

Abstract

The present invention relates to an antimicrobial composition for inhibiting antibiotic-resistant bacteria comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an active ingredient. An antimicrobial composition for inhibiting antibiotic-resistant bacteria comprising the 7,10-epoxy-octadeca-7,9-dienoic acid of the present invention and an antibiotic as an active ingredient is used (Staphylococcus aureus, MDRSA) can effectively inhibit the growth of Staphylococcus aureus and the resistant microorganisms derived therefrom, irrespective of the type of antibiotics and antibiotics. Therefore, it can be usefully used in the related pharmaceutical, food and cosmetic industries which require inhibition of growth of antibiotic resistant bacteria.

Description

7,10-epoxy-octadeca-7,9-dienoic acid and antibiotics, which comprises 7,10-epoxyoctadeca-7,9-dionanoic acid and an antibiotic as an active ingredient.

The present invention relates to an antimicrobial composition for inhibiting antibiotic-resistant bacteria comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an active ingredient.

Furan fatty acids (F-acids) are a large group of fatty acids specified by the furan ring and include unbranched fatty acid chains with 9, 11, or 13 carbon atoms at one a-position, and a straight-chain alkyl group having 3 or 5 carbon atoms at the? -position. Usually, the two [beta] -position of the furan ring is substituted with one or two methyl residues or one of the other groups. Furan fatty acids with no substitution at all in the β-position of the furan ring were also found in the seed oil of Exocarpus cupressiformis. Furan fatty acids are widely distributed in nature as trace components of mammals, including plants, fish, amphibians, reptiles, microorganisms and humans.

The biological role of furan fatty acids in biological systems is not entirely known, but it is known that furan fatty acids are involved in a variety of important biological functions that act as antioxidants, antitumor agents and antithrombotic agents. Furan fatty acids in some fish contain up to 25% of liver lipids and accumulate during the spawning season, thus indicating a possible correlation between furan fatty acid and the fertilization process. Correlations between consumption of furan fatty acid-rich fish and coronary heart disease mortality have been confirmed in several studies [(Spiteller, 2005, Lipids 40 (8): 755-771), (Ishii et al. 1989, Chem Pharm Bull 37 (5): 1396-1398) (Graff et al., 1984, Biochim Biophys Acta 799 (2): 143-150), (Parker et al., 1977, J Med Chem 20 ): 781-791). Furan fatty acids have also been shown to have inhibitory effects on platelet aggregation and potential antitumor activity. Furan fatty acids prevent the oxidation of linoleic acid and are known to act as antioxidants in plants. Some studies have shown that furan fatty acid acts as a radical scavenger by forming dioxoenes by oxidation of the ring opening in the presence of linoleic acid as a co-substrate. Respectively.

The biosynthesis of furan fatty acids is complex and diverse. The precursor of the biosynthesis of most furan fatty acids is known as linoleic acid. Plants are known to synthesize the basic skeleton of furan fatty acids. However, studies conducted with radio-labeled feeding on fish have shown that fish can not synthesize the alkyl side chain and the furan ring of the furan fatty acid. Thus, furan fatty acids in fish are thought to originate from the diet, especially the algae. As a result, furan fatty acids are introduced into the body through foods such as vegetables and fish. Food-derived furan fatty acids are introduced into mammalian tissues and blood, particularly phospholipids, which may act as radical scavengers and inhibit platelet aggregation.

Staphylococcus aureus is a gram-positive, tuberous anaerobic bacterium that forms cell clumps (lumps) as it grows, and is generally present in the skin and nasal surface of healthy persons and livestock (Nair et al., 2015, Antimicrob Agents Chemother 59 (4): 1876-1885). It produces exothermic exotoxin causing food poisoning. It also releases toxins (luocosidine), hemolysin, and coagulase enzymes that kill phagocytes, causing it to escape the resistance of infected host cells and cause pyogenic infections. Recently, MRSA (methicillin-resistant Staphylococcus aureus), a strain showing resistance to most antibiotics, has appeared in hospitals (David and Daum, 2010, Clin Microbiol Rev 23 (3): 616-687). The MRSA is resistant to antibiotics such as penicillin, which has been used for about 50 years as a prescription medicine for bacterial infections. 2006), Lancet (2006), and the Lancet (2006), who have been hospitalized in hospitals and medical institutions, and are infected between humans and humans through physical contact, resulting in post-operative trauma or pneumonia 368 (9538): 874-885). Although the various physiological activities of furan fatty acid are well known, the antimicrobial activity against antibiotic resistant bacteria and the antibacterial effect against the antibiotic combination treatment have not yet been confirmed.

Accordingly, the present inventors have searched for a method having effective antimicrobial activity against antibiotic-resistant bacteria, and found that when 7,10-epoxyoctadeca-7,9-dionanoic acid and antibiotics are used in combination, they inhibit resistant bacteria derived from Staphylococcus aureus And thus the present invention has been completed.

Accordingly, an object of the present invention is to provide a method for inhibiting antibiotic-resistant bacteria containing 7,10-epoxy-octadeca-7,9-dienoic acid and an antibiotic as an active ingredient. To provide an antimicrobial composition.

It is also an object of the present invention to provide a method of killing antibiotic resistant bacteria comprising treating 7,10-epoxyoctadeca-7,9-diono acid and an antibiotic.

It is also an object of the present invention to provide a pharmaceutical composition for preventing or treating antibiotic-resistant infectious disease comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an active ingredient.

It is also an object of the present invention to provide an antimicrobial cosmetic composition for antibiotic resistant bacteria comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an active ingredient.

It is also an object of the present invention to provide an antimicrobial food composition for antibiotic resistant bacteria comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an active ingredient.

It is also an object of the present invention to provide an antimicrobial animal feed composition for antibiotic resistant bacteria comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an active ingredient.

It is also an object of the present invention to provide a disinfectant for antibiotic resistant bacteria comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an active ingredient.

It is also an object of the present invention to provide an antimicrobial adjuvant for the killing of antibiotic-resistant bacteria containing 7,10-epoxyoctadeca-7,9-dienoic acid as an active ingredient.

In order to solve the above-mentioned problems, the present invention provides a pharmaceutical composition containing 7,10-epoxy-octadeca-7,9-dienoic acid and an antibiotic as an active ingredient And an antibiotic-resistant microorganism.

The present invention also provides a method of killing antibiotic resistant bacteria comprising treating 7,10-epoxyoctadeca-7,9-diono acid and an antibiotic.

The present invention also provides a pharmaceutical composition for preventing or treating antibiotic-resistant microbial infectious diseases comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an active ingredient.

The present invention also provides an antimicrobial cosmetic composition for antibiotic resistant bacteria comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an active ingredient.

The present invention also provides an antimicrobial food composition for antibiotic resistant bacteria comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an effective ingredient.

In addition, the present invention provides an antimicrobial animal feed composition for antibiotic resistant bacteria comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an effective ingredient.

The present invention also provides a disinfectant for antibiotic-resistant bacteria comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an effective ingredient.

The present invention also provides an antimicrobial adjuvant for the killing of antibiotic resistant bacteria comprising 7,10-epoxyoctadeca-7,9-dienoic acid as an active ingredient.

An antimicrobial composition for inhibiting antibiotic-resistant bacteria comprising the 7,10-epoxy-octadeca-7,9-dienoic acid of the present invention and an antibiotic as an active ingredient is used (Staphylococcus aureus, MDRSA) can effectively inhibit the growth of Staphylococcus aureus and the resistant microorganisms derived therefrom, irrespective of the type of antibiotics and antibiotics. Therefore, it can be usefully used in the related pharmaceutical, food and cosmetic industries which require inhibition of growth of antibiotic resistant bacteria.

Figure 1 is a schematic representation of a process for preparing 7,10-epoxyoctadeca-7,9-dienoic acid (7,10-dioctyl-8,10-dihydroxy- -EODA) and 7,10-epoxyoctadeca-7,9-dienoic acid.
FIG. 2 (1) compares the antimicrobial activity of the 7,10-EODA or the antibiotic of the present invention against the MDRSA 01ST001 strain (S is 100 μg of streptomycin, A is 100 μg of ampicillin, O is oxacillin 100 μg, V means vancomycin 100 μg, P means penicillin 100 μg, and E means 7,10-EODA 100 μg respectively). (V is 100 μg of vancomycin, O is 100 μg of oxacillin, E1 is 7,10-EODA of 50 μg, E2 is 7, 10-EODA, 10-EODA 100 μg, E3 7,10-EODA 200 μg, and E4 7,10-EODA 400 μg, respectively). (3) and (4) compare antimicrobial activity of 7,10-EODA of the present invention with oxacillin (3) and ampicillin (4) after combination treatment (a is 5 μg of each antibiotic alone, B was 100 μg of each antibiotic and 31.2 μg of 7,10-EODA, and S was 7,10-EODA of 31.2 μg. A, 100 μg of antibiotic alone, 5 μg of each antibiotic and 31.2 μg of 7,10- Meaning).
FIG. 3 shows the result of confirming the synergistic effect of antimicrobial activity according to the treatment concentration in the combination treatment of penicillin and 7,10-EODA of MDRSA 01ST001 strain according to the present invention (where? Is the untreated 7,10-EODA,? Treatment of 7.5 μg / mL of 7,10-EODA, and ▼ treatment of 15.0 μg / mL of 7,10-EODA).
FIG. 4 shows the results of confirming the growth of the strain MDRSA 01ST001 according to the incubation time and 7,10-EODA and the antibiotics combination treatment (in the untreated group, ○ indicates 15.6 μg concentration (1/8 × MIC) , 10-EODA treated group (a), ▼ was treated with 7,10-EODA treated group with a concentration of 31.2 μg (1/4 × MIC), Δ was treated with oxacillin (c) at a concentration of 3.9 μg, and 15.6 μg (A + c) in the concentration of 7,10-EODA + 3.9 μg and the oxysilane-treated group (b + c) in the concentration of 31.2 μg in the 7,10-EODA + 3.9 μg Meaning).
FIG. 5 shows the result of confirming the synergistic effect of antimicrobial activity against MRSA or MSSA strain in combination with 7,10-EODA and non-beta-lactam (MRSA 01ST 018) ,? Means MDRSA 01ST 001,? Means S. aureus ATCC 29213, and? Means S. aureus 1199).

The present invention provides an antimicrobial composition for inhibiting antibiotic-resistant bacteria comprising 7,10-epoxy-octadeca-7,9-dienoic acid and an antibiotic as an active ingredient do.

In the present invention, the term " 7,10-epoxy-octadeca-7,9-dienoic acid " 1 is a compound represented by the following formula (1), which is linked with an epoxy structure to form a furan ring composed of carbon atoms 7, 8, 9 and 10.

[Chemical Formula 1]

The 7,10-epoxyoctadeca-7,9-dienoic acid (7,10-EODA) may be chemically synthesized or obtained in nature, and in one embodiment of the present invention 7,10-dihydroxy-8 (E) -octadecenoic acid (DOD) is used.

Specifically, the 7,10-epoxyoctadeca-7,9-dienoic acid (EODA) was prepared by adding hexane to 7,10-dihydroxy-8 (E) -octadecenoic acid (DOD) (See Fig. 1).

The 7,10-dihydroxy-8 (E) -octadecenoic acid (DOD) is a type of hydroxy fatty acid, and has a total of two hydroxyl groups, one in each of the 7-carbon and 10- , And one trans double bond between carbon number 8 and carbon number 9.

The mixing ratio of the 7,10-dihydroxy-8 (E) -octadecenoic acid (DOD) and hexane may be appropriately selected, but 7,10-dihydroxy-8 (E) -octadecenoic acid (DOD (EOD) octadecenoic acid (DOD), which is used to facilitate the smooth mixing and the reaction of 7,10-dihydroxy-8 (E) More preferably 10 to 1000 μl of hexane per 10 mg of 7,10-dihydroxy-8 (E) -octadecenoic acid (DOD).

Preferably, the heat treatment is performed at 30 to 150 ° C for 1 to 150 hours, more preferably at 90 to 90 ° C, 95 ° C for 12 to 96 hours.

In the present invention, antibiotics usable in combination with 7,10-epoxyoctadeca-7,9-dienoic acid are beta-lactam-based or non-beta-lactam antibiotics and are not limited thereto.

 In the present invention, the term " beta-lactam "means a heteroatomic ring consisting of three carbon atoms and one nitrogen atom. Beta-lactam rings are common to many antibiotics, and they are called beta-lactam antibiotics. Beta-lactam antibiotics inhibit bacterial cell wall synthesis and exhibit antibacterial activity.

The non-beta-lactam antibiotic refers to an antibiotic that inhibits protein synthesis, inhibits DNA / RNA synthesis, and exhibits antimicrobial activity by cell membrane-destroying activity.

The beta-lactam antibiotic is one or more selected from the group consisting of penicillin, Cephalosporin, Carbapenem, Monobactam, and vancomycin, But is not limited thereto.

The penicillin antibiotics may be selected from the group consisting of benzylpenicillin, penoxymethylpenicillin, ampicillin, amoxycillin, bacampicillin, methicillin, oxacillin, Oxacillin, Cloxacillin, Piperacillin, Carbenicillin, and Ticarcillin. The antibiotics of the present invention are not limited thereto.

The cephalosporin antibiotic is selected from the group consisting of Cephradine, Cefadroxil, Cephalexin, Cefazolin, Cephapirin, Cefaclor, Cefaclor, Which consists of Cefuroxime, Cefamandole, Cefotiam, Cefoxitin, Cefixime, Cefpodoxime, Cefotaxime and Ceftriaxone. But not limited thereto.

The carbapenem antibiotic is at least one selected from the group consisting of Tienam, Meropenem, imipenem, Doripenem and Ertapenem, It does not.

The antibiotic of the vancomycin family is not limited to vancomycin or teicoplanin.

The non-beta-lactam antibiotic may be selected from the group consisting of aminoglycoside, tetracycline, macrolide, quinolone, rifampicin, sulphonamides, But are not limited to, one selected from the group consisting of Polypeptide series, Lincosamide series, Fluoroquinolone series and Cephalosporin series.

The aminoglycoside antibiotic may be selected from the group consisting of amikacin, sisomycin, arbekacin, gentamycin, kanamycin, neomycin, spectinomycin, But are not limited to, one or more selected from the group consisting of spectinomycin, netilmicin, paromomycin, streptomycin and tobramycin.

The tetracycline antibiotic may be at least one selected from the group consisting of Chlortetracycline, Oxytetracycline, Tetracycline, Minocycline and Doxycycline. It does not.

The macrolide antibiotic is one or more selected from the group consisting of Erythromycin, Spiramycin, and Tylosin, but is not limited thereto.

The quinolone antibiotic is one or more selected from the group consisting of nalidixic acid, Moxifloxacin, Ofloxacin, trovan and levofloxacin, But is not limited thereto.

The rifampicin-based antibiotic is rifamycin, rifampicin or rifaximin, but is not limited thereto.

The above sulfonamides antibiotics include sulfamethazine, sulfadimidine, sulfamethoxazine, sulfamethoxazole, sulfaguanidine, sulfapyridine, sulfamethoxazine, sulfamethoxazine, sulfamethoxazine, sulfamethoxazine, But is not limited to, at least one selected from the group consisting of single catechol and sulfathiazole.

The Polypeptide antibiotic is Polymyxin B or Bacitracin, but is not limited thereto.

The Lincosamide family antibiotic is Lincomycin or Clindamycin, but not limited thereto.

The fluoroquinolone antibiotic is selected from the group consisting of Norfloxacin, Ciprofloxacin, Enrofloxacin, Ofloxacin and pefloxacin. But is not limited thereto.

Cephalosporin antibiotics include Cephalexin, Cefradine, Cefazolin, Cephaloridine, Cefaclor, Ceftezole, Sephalanol, Cephalosporin, Consisting of Cefamandole, Cefuroxime, Cefoxitin, Cefpiramide, Cefoperazone, Moxalactam, Ceftiofur, and Cefuzonam. But not limited thereto.

The antimicrobial composition has an antibacterial effect on both antibiotic susceptible bacteria and antibiotic resistant bacteria. In particular, the antimicrobial composition of the present invention is more effective for antibiotic resistant bacteria, and thus it is possible to treat infectious diseases which are difficult to treat with antibiotics.

The antibiotic-resistant bacteria may be selected from the group consisting of Staphylococcus aureus , Escherichia coli , Salmonella enteritidis , Streptococcus pyogenes , Pseudomonas aeruginosa, aeruginosa , and Clostridium difficile . Preferably, the staphylococcus aureus is not limited to Staphylococcus aureus.

In the present invention, the term " Staphylococcus aureus " is a gram-positive, tuberous anaerobic bacterium that forms cell clumps (lumps) as it grows, and is generally present on the skin and nasal surface of healthy persons or livestock . Produces heat-resistant exotoxin, causing food poisoning, releasing toxins (luocosidine), hemolysin, coagulation enzymes, etc., which cause phagocytic infections by deviating from the resistance of infected host cells. Recently, antibiotic-resistant bacteria showing resistance to most antibiotics have appeared in hospitals and the like, which is causing serious social problems.

The Staphylococcus aureus is a multidrug-resistant Staphylococcus aureus (Staphylococcus aureus) (multi-drug resistant Staphylococcus aureus, MDRSA), methicillin-resistant Staphylococcus aureus (methicillin-resistant staphylococcus aureus, MRSA ), vancomycin-resistant Staphylococcus aureus (vancomycin resistant staphylococcus aureus, VRSA), and vancomycin intermediate Staphylococcus aureus (VISA), and more preferably at least one selected from the group consisting of Staphylococcus aureus (multi-drug resistant Staphylococcus aureus, MDRSA ) Or methicillin-resistant staphylococcus aureus (MRSA).

In the present invention, the "multidrug-resistant Staphylococcus aureus (Staphylococcus aureus) (multidrug-resistant Staphylococcus aureus, MDRSA)" is a Staphylococcus aureus resistant to a wide variety of series, as well as methicillin antibiotics, As many a variety of antibiotics It is a bacterium that is resistant to most antibiotics and can only be treated with an extremely limited antibiotic.

In the present invention, the term " methicillin-resistant staphylococcus aureus (MRSA) "is a Staphylococcus aureus resistant to an antibiotic of methicillin, a penicillin antibiotic, It is the most frequent causative bacterium, and it is fatal to the patient who has decreased immunity or the elderly.

It is obvious that the concentration of 7,10-epoxyoctadeca-7,9-dienoic acid (EODA) contained in the antimicrobial composition can be variously selected according to the target microorganism, and preferably 0.01 to 5,000 μg / ml. < / RTI > If the concentration is less than 0.01 μg / ml, the antimicrobial activity may not be exhibited, and if it exceeds 5,000 μg / ml, it may be toxic to the human body.

In one embodiment of the present invention, the 7,10-EODA of the present invention can be used in combination with other antibiotics to prevent antagonism without antagonism, And the antimicrobial activity synergistic effect was shown. In addition, the 7,10-EODA of the present invention is excellent in antimicrobial activity and growth inhibitory effect in combination with various antibiotics besides beta-lactam antibiotics, and can be usefully used in the related pharmaceutical, food and cosmetic industries.

The present invention also provides a method of killing antibiotic resistant bacteria comprising treating 7,10-epoxyoctadeca-7,9-diono acid and an antibiotic.

The antimicrobial method is a combination treatment of the 7,10-epoxyoctadeca-7,9-dionanoic acid and the antibiotic of the present invention, and the sequence of the antibiotic is not limited. Also, the method of death is not limited as long as the 7,10-epoxyoctadeca-7,9-dionanoic acid and the antibiotic can be treated at the same time or can be treated individually and sequentially, and the object of destroying the antibiotic-resistant bacteria can be achieved .

The present invention also provides a pharmaceutical composition for preventing or treating antibiotic-resistant microbial infectious diseases comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an active ingredient.

The antibiotic-resistant microorganism infectious disease is at least one selected from the group consisting of abscess, dermatitis, osteoarthritis, bacteremia, pneumonia, toxic shock syndrome, food poisoning, high fever, sepsis, edema disease, mastitis and intestinal colicosis.

The pharmaceutical compositions of the present invention may further comprise adjuvants other than 7,10-epoxyoctadeca-7,9-dionanoic acid and antibiotics. Such adjuvants may be used without limitation as long as they are known in the art, but may include, for example, Freund's complete or incomplete adjuvant to increase its immunity.

The pharmaceutical composition according to the present invention can be prepared by incorporating the active ingredient into a pharmaceutically acceptable carrier. Here, the pharmaceutically acceptable carrier includes carriers, excipients and diluents commonly used in the pharmaceutical field. Pharmaceutically acceptable carriers for use in the pharmaceutical compositions of the present invention include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin , Calcium phosphate, calcium silicate, cellulose, methylcellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The pharmaceutical composition of the present invention can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols or the like, oral preparations, suppositories or sterilized injection solutions according to a conventional method .

In the case of formulation, it can be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants and the like which are usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid preparations 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 may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to commonly used diluents such as water and liquid paraffin, various excipients such as wetting agents, sweetening agents, fragrances and preservatives . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. Propellants, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like can be used. As the base of suppositories, witepsol, tween 61, cacao paper, laurin, and glycerogelatin can be used.

The pharmaceutical composition according to the present invention can be administered to the individual by various routes. All modes of administration may be expected, for example, by oral, intravenous, intramuscular, subcutaneous, intraperitoneal injection.

The dosage of the pharmaceutical composition according to the present invention is selected in consideration of the age, weight, sex, physical condition, etc. of the individual. It is obvious that the concentration of 7,10-epoxyoctadeca-7,9-dienoic acid (EODA) contained in the pharmaceutical composition can be variously selected depending on the subject, and preferably 0.01 to 5,000 占 퐂 / ml. < / RTI > If the concentration is less than 0.01 μg / ml, the pharmaceutical activity may not be exhibited. If the concentration is more than 5,000 μg / ml, it may be toxic to the human body.

The present invention also provides an antimicrobial cosmetic composition for antibiotic resistant bacteria comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an active ingredient.

The composition comprising the 7,10-epoxyoctadeca-7,9-dionanoic acid and the antibiotic of the present invention can be used variously for preventing or improving antibiotic-resistant infectious disease. Examples of products to which the present composition can be added include cosmetics such as various creams, lotions, skins, essences and the like and shampoos, rinses, cleansers, cleansers, soaps, treatments, packs and essences.

The cosmetic composition of the present invention comprises a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymeric polysaccharides, sphingolipids and seaweed extracts.

The water-soluble vitamin is not particularly limited as long as it can be compounded in cosmetics. Preferably, vitamin B, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C, And their salts (thiamine hydrochloride, sodium ascorbate, etc.) or derivatives (sodium ascorbic acid-2-phosphate, magnesium ascorbate-2-phosphate etc.) can also be added to water-soluble vitamins . The water-soluble vitamin can be obtained by a conventional method such as a microorganism conversion method, a purification method from a culture of a microorganism, an enzymatic method, or a chemical synthesis method.

Usable vitamins include vitamins such as vitamin A, carotene, vitamin D2, vitamin D3, vitamin E (d1-alpha tocopherol, d-alpha tocopherol, d-alpha tocopherol) Alpha-tocopherol vitamin E, DL-pantothenyl alcohol, D-pantothenyl alcohol, pantothenyl ethyl laurate, and the derivatives thereof (palmitic acid ascorbin, stearic acid ascorbic acid, dipalmitic acid ascorbin, dl-alpha tocopherol acetate, Ether, etc.) are also included in the usable vitamins used in the present invention. Usability Vitamins can be obtained by a conventional method such as a microorganism conversion method, a purification method from a culture of a microorganism, an enzyme or a chemical synthesis method.

The polymeric peptide may be any compound as long as it can be compounded in cosmetics, and examples thereof include collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, and keratin. The polymeric peptide can be obtained by a conventional method such as purification from a culture broth of a microorganism, an enzymatic method, or a chemical synthesis method, or it can be purified from natural products such as ducks such as pigs and cows and silk fiber of silkworms.

The polymeric polysaccharide may be any compound as long as it can be incorporated in cosmetics, and examples thereof include hydroxyethyl cellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate or a salt thereof (sodium salt, etc.). For example, chondroitin sulfate or a salt thereof can be usually purified from mammals or fish.

Sphingo lipids may be any as long as they can be incorporated into cosmetics, and preferable examples thereof include ceramides, phytosphingosine and sphingoglycolipids. Sphingoid lipids can be purified from ordinary mammals, fish, shellfish, yeast or plants by conventional methods or can be obtained by chemical synthesis.

The seaweed extract may be any of those which can be compounded in cosmetics. Preferably, the seaweed extract is selected from the group consisting of algae extract, red pepper extract, green algae extract, and the like. Potassium alginate and the like are also included in the seaweed extract used in the present invention. Seaweed extract can be obtained from seaweed by a conventional method.

The cosmetic of the present invention may be blended with other essential ingredients, if necessary, in combination with the essential ingredients. Examples of the compounding ingredients that may be added include organic solvents such as a preservative component, a moisturizer, an emollient, a surfactant, an organic and inorganic pigment, an organic powder, an ultraviolet absorbent, a preservative, a bactericide, an antioxidant, a plant extract, a pH adjuster, A blood circulation accelerator, a cold agent, an antiperspirant agent, and purified water. Examples of the oil retaining component include ester-based oil retaining, hydrocarbon-based oil retaining, silicone-based oil retaining, fluoric oil retaining, animal retention and plant retention.

Examples of ester-based fats include glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isostearyl isostearate, Butyl isopropyl myristate, isopropyl myristate, isopropyl myristate, isopropyl myristate, isopropyl myristate, isopropyl myristate, butyl, ethyl linoleate, isopropyl linoleate, ethyl oleate, isosilyl myristate, isostearic acid isostearyl, isostearyl palmitate, octyldodecyl myristate, Trimethylol propane, triisostearic acid trimethylol propane, tetra 2-ethylhexanoic acid pentaerythritol tetra (2-ethylhexanoate) , Decyl caprylate, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, myristine acid stearate, stearyl stearate, decyl oleate, ricinoleic acid Terephthalic acid, isostearyl stearate, isocetyl stearate, isodecyl stearate, octyldodecyl oleate, octyldodecyl linoleate, isopropyl stearate, isostearyl stearate, isostearyl stearate, Ethylhexanoate, stearyl 2-ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, di (capryl, capric acid) propylene glycol, Propylene glycol dicaprylate, propylene glycol dicaprylate, neopentyl glycol dicaprate, neopentyl glycol dioctanoate, glyceryl tricarboxylate, glyceryl triunsaturated acid, glyceryl triisopalmitate, glyceryl triisostearate, octyl neopentanoate Dodecylsuccinic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, , Octyldecyl isostearate, polyglycerin oleic acid ester, polyglycerin isostearic acid ester, triisocetyl citrate, triisolealkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, citric acid But are not limited to, triethyl citrate, acetyltriethyl citrate, acetyl tributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di-2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate , Dodecyl sebacate, stearic acid cholesteryl, isostearic acid cholesteryl, hydroxystearic acid cholesteryl, oleic acid cholesteryl, oleic acid dihydrocholesteryl, isostearic acid pitostearyl, oleic acid piteste Stearyl hydroxystearate, stearyl 12-stearoyl hydroxystearate, 12-stearoyl hydroxystearic acid, Allo one hydroxy stearic acid and the like esters such as cetearyl source.

Hydrocarbon-based fats and oils include hydrocarbon-based fats such as squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, floating isoparaffin, polybutene, microcrystalline wax and vaseline.

Examples of silicone based oils include polymethyl silicone, methylphenyl silicone, methyl cyclopolysiloxane, octamethyl polysiloxane, decamethyl polysiloxane, dodecamethyl cyclosiloxane, dimethyl siloxane and methyl cetyloxysiloxane copolymer, dimethyl siloxane and methyl stearoxysiloxane copolymer, alkyl Modified silicone oils, and amino-modified silicone oils.

Examples of the fluorine-based oil include perfluoropolyether and the like.

Examples of animal or vegetable oils include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, new flower oil, soybean oil, corn oil, rape oil, apricot kernel oil, palm kernel oil, palm oil, castor oil, , Corn oil, palm oil, palm oil, cucumber nut oil, wheat germ oil, rice germ oil, shea butter, coltsfoot colostrum, marker daisy nut oil, mead home oil, egg oil, , Canned wax, carnauba wax, liquid lanolin, hardened castor oil, and the like.

Examples of the moisturizing agent include water-soluble low-molecular moisturizing agents, oil-soluble molecular moisturizing agents, water-soluble polymers, and oil-soluble polymers.

Examples of the water-soluble low-molecular moisturizing agent include serine, glutamine, sorbitol, mannitol, sodium pyrrolidone-carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B Glycol (polymerization degree n = 2 or more), polyglycerin B (polymerization degree n = 2 or more), lactic acid, lactic acid salt and the like.

Examples of the lipid-soluble low-molecular moisturizing agent include cholesterol and cholesterol ester.

Examples of the water-soluble polymer include carboxyvinyl polymer, polyaspartic acid, tragacanth, xanthan gum, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, water-soluble chitin, chitosan, dextrin, etc. .

Examples of the oil-soluble polymer include polyvinyl pyrrolidone and eicosene copolymers, polyvinyl pyrrolidone and hexadecene copolymers, nitrocellulose, dextrin fatty acid esters, and polymer silicon. Examples of the emollients include long chain acyl glutamic acid cholesteryl ester, hydroxystearic acid cholesteryl, 12-hydroxystearic acid, stearic acid, rosin acid and lanolin fatty acid cholesteryl ester.

Examples of the surfactant include nonionic surfactants, anionic surfactants, cationic surfactants, and amphoteric surfactants.

Examples of nonionic surfactants include self emulsifying monostearic acid glycerin, propylene glycol fatty acid esters, glycerin fatty acid esters, polyglycerin fatty acid esters, sorbitan fatty acid esters, POE (polyoxyethylene) sorbitan fatty acid esters, POE sorbit fatty acid esters, Polyoxyethylene and polyoxypropylene) copolymers, POE and POP alkyl ethers, polyether modified silicones, polyether modified polyesters, polyoxyethylene polyoxypropylene (POE) fatty acid esters, POE alkyl ethers, POE fatty acid esters, POE hardened castor oil, POE castor oil, POE and POP Acid alkanolamides, alkylamine oxides, hydrogenated soybean phospholipids, and the like.

Examples of anionic surfactants include fatty acid soap, alpha-acylsulfonate, alkylsulfonate, alkylarylsulfonate, alkylnaphthalenesulfonate, alkylsulfate, POE alkyl ether sulfate, alkylamide sulfate, alkyl phosphate, POE alkyl, Alkylsulfosuccinic acid salts, acylated hydrolyzed collagen peptide salts, and perfluoroalkyl phosphoric acid esters, and the like can be mentioned. have.

Examples of the cationic surfactant include alkyl trimethyl ammonium chloride, stearyl trimethyl ammonium chloride, stearyl trimethyl ammonium bromide, cetostearyl trimethyl ammonium chloride, distearyl dimethyl ammonium chloride, stearyl dimethyl benzyl ammonium chloride, behenyl trimethyl ammonium chloride, Benzalkonium, diethylaminoethylamide stearate, dimethylaminopropylamide stearate, quaternary ammonium salts of lanolin derivatives, and the like.

Examples of the amphoteric surfactant include carboxybetaine type, amide betaine type, sulfobetaine type, hydroxysulfobetaine type, amidosulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type and amide amine type Amphoteric surfactants and the like.

Examples of the organic and inorganic pigments include inorganic pigments such as silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, Bengala, clay, bentonite, titanium mica, titanium oxide, bismuth chloride, zirconium oxide, magnesium oxide, Inorganic pigments such as calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, chromium oxide, chromium oxide, chromium hydroxide, Polyurethane, vinyl resin, urea resin, phenol resin, fluorine resin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene and styrene copolymer, Silk powder, cellulose, CI Pigment Yellow, CI Pigment Orange, and composite pigments of inorganic pigments and organic pigments thereof.

As the organic powder, metallic soap such as calcium stearate; Metal salts of alkyl phosphates such as sodium zinc cetylate, zinc laurylate and calcium lauryl laurate; Acylamino acid polyvalent metal salts such as N-lauroyl-beta-alanine calcium, N-lauroyl-beta-alanine zinc and N-lauroylglycine calcium; Amidosulfonic acid multivalent metal salts such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; Such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoylidene, N-alpha-paratyylnitine, N-alpha-lauroyl arginine, Acyl basic amino acids; N-acylpolypeptides such as N-lauroylglycylglycine; Alpha-amino fatty acids such as alpha-aminocaprylic acid, alpha-aminoaurauric acid, and the like; Polyethylene, polypropylene, nylon, polymethylmethacrylate, polystyrene, divinylbenzene and styrene copolymers, and ethylene tetrafluoride.

Examples of ultraviolet absorbers include paraaminobenzoic acid, ethyl parnamobenzoate, amyl paranobenzoate, octyl paranobenzoate, ethyleneglycol salicylate, phenyl salicylate, benzyl salicylate, benzyl salicylate, butylphenyl salicylate, homomenthyl salicylate, benzyl cinnamate , 2-ethoxyethyl para-methoxy cinnamate, octyl paratomethoxy cinnamate, divaramethoxy cinnamoamnoso-2-ethylhexane glyceryl, isomethoxy methacrylate, diisopropyl and diisopropyl cinnamate ester mixtures, Dihydroxymethoxybenzophenone sulfonic acid, dihydroxybenzophenone sulfonic acid, dihydroxybenzophenone sulfonic acid and its salt, dihydroxymethoxybenzophenone, sodium dihydroxymethoxybenzophenone disulfonate, dihydroxybenzo Phenanone, tetrahydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianylino-p- (carbo-2'-ethylhexyl- 1,3,5-triazine, 2- (2-H And the like can be mentioned hydroxy-5-methylphenyl) benzotriazole.

Examples of the disinfectant include hinokitiol, trichloroacid, trichlorohydroxydiphenyl ether, crohexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, zinc filitione, benzalkonium chloride, No. 301, mononitro and eicol sodium, and undecylenic acid.

Examples of the antioxidant include butylhydroxyanisole, gallic acid propyl, and eicosorbic acid.

Examples of the pH adjuster include citric acid, sodium citrate, malic acid, sodium malate, fumaric acid, sodium fumarate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogenphosphate and the like.

Examples of the alcohol include higher alcohols such as cetyl alcohol.

The components to be added in addition to these components are not limited thereto, and any of the above components can be compounded within a range that does not impair the objects and effects of the present invention.

The cosmetic of the present invention may take the form of a solution, an emulsion, a viscous mixture or the like.

The ingredients contained in the cosmetic composition of the present invention may include the ingredients conventionally used in cosmetic compositions as an active ingredient and may contain conventional auxiliary agents such as stabilizers, solubilizers, vitamins, pigments and flavoring agents, and carriers .

The cosmetic composition for preventing skin whitening and skin aging according to the present invention may be prepared in any formulations conventionally produced in the art and may be formulated into various forms such as emulsion, cream, lotion, pack, foundation, lotion, essence, .

Specifically, the cosmetic composition of the present invention can be used as a skin lotion, a skin softener, a skin toner, a milk lotion, an astringent, a lotion, a moisturizing lotion, a nutrition lotion, a massage cream, a nutrition cream, a moisturizing cream, a hand cream, A lotion, a cleansing cream, a hair lotion, a hair tonic, a hair essence, a hair shampoo, a hair rinse, a hair treatment, a body lotion and a body cleanser.

When the formulation of the present invention is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.

In the case of the solution or emulsion of the present invention, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

When the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic derivatives or ethoxylated glycerol fatty acid esters.

The present invention also provides an antimicrobial food composition for antibiotic resistant bacteria comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an effective ingredient.

7,10-Epoxyoctadeca-7,9-dionanoic acid and antibiotics of the present invention can be used as health functional foods, food additives or dietary supplements.

When the 7,10-epoxyoctadeca-7,9-dienoic acid and the antibiotic are used as a food additive, they may be added as they are, mixed with other food or food ingredients, and used appropriately according to a conventional method .

The mixing amount of the 7,10-epoxyoctadeca-7,9-dionanoic acid and the antibiotic can be suitably changed according to the intended use (prevention, health or therapeutic treatment) By weight, preferably 0.01 to 95% by weight, more preferably 0.1 to 80% by weight. If the content is less than 0.01% by weight, the efficiency of taking may be lowered. If the content is more than 80% by weight, formulation may be difficult.

As a specific example, in the production of food or beverage, the 7,10-epoxyoctadeca-7,9-dioic acid and the antibiotic of the present invention are added in an amount of not more than 15% by weight, preferably not more than 10% do. However, when it is intended for health and hygiene purposes or for the purpose of controlling health, it can be added in an amount below the above range, and there is no problem in terms of safety. Therefore, the active ingredient can be used in an amount exceeding the above range have.

There is no particular limitation on the kind of the food, but examples of the food to which the 7,10-epoxyoctadeca-7,9-dienoic acid and the antibiotic of the present invention can be added include meat, sausage, bread, chocolate, candy, Dairy products including ice cream, various soups, drinks, tea, drinks, alcoholic beverages, and vitamin complexes, all of which include health foods in the conventional sense.

When the food composition of the present invention is prepared as a beverage, it may contain additional ingredients such as various flavors or natural carbohydrates such as ordinary beverages. Examples of the natural carbohydrate include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Natural sweetening agents such as dextrin and cyclodextrin; Synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The natural carbohydrate is contained in an amount of 0.01 to 10% by weight, preferably 0.01 to 0.1% by weight based on the total weight of the food composition of the present invention.

The food composition of the present invention can be used for various foods, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickening agents, pH adjusters, stabilizers, preservatives, glycerin, Carbonation agents used in beverages, etc., and may include, but is not limited to, natural fruit juices, fruit juice beverages, and flesh for the manufacture of vegetable beverages. These components may be used independently or in combination. The proportion of the above additives is not particularly limited, but is preferably within a range of 0.01 to 0.1% by weight based on the total weight of the food composition of the present invention.

For long-term consumption intended for health and hygiene purposes or health control purposes, the food composition of the present invention has no problem in terms of safety and can be taken for a long period of time.

In addition, the present invention provides an antimicrobial animal feed composition for antibiotic resistant bacteria comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an effective ingredient.

The 7,10-epoxyoctadeca-7,9-dienoic acid and the antibiotic are preferably contained in an amount of 0.01 to 95% by weight, more preferably 1 to 80% by weight, based on the total weight of the animal feed composition will be. If the content is less than 0.01% by weight, the efficiency of taking may be poor. If the content is more than 95% by weight, formulation may be difficult.

The animal feed composition of the present invention comprising 7,10-epoxyoctadeca-7,9-dionanoic acid and an antibiotic as an active ingredient can prevent or ameliorate an infectious disease caused by a pathogenic bacterium in an animal by feeding to livestock .

The animal to which the animal feed composition is administered may be both ruminant and non-ruminant, but preferably it may be a ruminant, and the species may be cattle, camels, deer, giraffe, and the like.

The animal feed composition of the present invention may further contain an organic acid such as citric acid, fumaric acid, adipic acid, lactic acid, and malic acid, sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polymerized phosphate) And natural antioxidants such as polyphenol, catechin, alpha-tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid and phytic acid.

In addition, the animal feed composition may further contain auxiliary components such as amino acids, inorganic salts, vitamins, antibiotics, antibacterials, antioxidants, antifungal enzymes, living microbial preparations and the like. Specifically, the auxiliary component may be selected from the group consisting of cereals (for example, ground or crushed wheat, oats, barley, corn, rice, etc.), vegetable protein feeds (for example, (For example, animal fat, vegetable fat, etc.), animal protein feeds (e.g. blood, meats, bone meal, fish meal etc.), sugar and dairy products (e.g., dry ingredients comprising various powdered milk or whey powders) ), Nutritional supplements, digestion and absorption enhancers, growth promoters, disease prevention agents, etc.

The animal feed composition may be administered alone to an animal or may be administered in admixture with other feed additives in an edible carrier. In addition, the animal feed composition may be mixed with the top dressing of the feed, or may be easily administered in combination with a feed or a separate additive or a separate oral formulation. The dosage of the animal feed composition can be suitably adjusted by using a single daily intake or a divided daily intake administered in the art.

The present invention also provides a disinfectant for antibiotic-resistant bacteria comprising 7,10-epoxyoctadeca-7,9-dienoic acid and an antibiotic as an effective ingredient.

The present invention also provides an antimicrobial adjuvant for the killing of antibiotic resistant bacteria comprising 7,10-epoxyoctadeca-7,9-dienoic acid as an active ingredient.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention as defined by the appended claims. It will be obvious to you.

Experimental Example  One. 7,10-Epoxy- Octadeca -7,9- Dienic acid (7,10-epoxy octadeca -7,9-dienoic acid, EODA)

10 mg of 7,10-dihydroxy-8 (E) -octadecenoic acid (DOD) and 500 μl of hexane as a solvent were placed in 4 ml glass vials and incubated in a heating block (Barnstead / Thermolyne Type 176000 Dri-Bath). After completion of the reaction, the solvent was removed using nitrogen flushing, and the reaction product was dissolved in a mixture of chloroform and methanol (1: 1, v / v) to obtain a product.

The resultant product was confirmed by thin layer chromatography, gas chromatography, gas chromatography / mass spectrometry and the like. Thin layer chromatography was performed on a glass substrate coated with silica gel. The solvent for the separation of the product was a mixture of Toluene: Dioxan: Acetic acid (79: 14: 7, v / v / v) Respectively. After the product was separated, the product was sprayed on the substrate with 50% sulfuric acid solution and heated at 100 ° C for more than 10 minutes. Gas chromatographic analysis was carried out by adding 1 ml of diazomethane to 10 mg of the sample to be analyzed and then leaving at room temperature for 5 minutes. The diazomethane was removed by using nitrogen gas, and 1 ml of TMSI + pyridine (1: 4, v / v) was added and left for 40 minutes. After the residual solvent was blown off using nitrogen gas, 200 μl of a solution for gas chromatography (Dichrolomethane: Methanol = 95: 5, v / v) was added. 1 μl of the final sample thus prepared was injected into a gas chromatograph analyzer. At this time, the column used was a hydrophobic column, the analysis temperature was in the range of 100 to 300, and the analysis time was within 1 hour. The analysis of gas chromatography / mass spectrometer was carried out according to the above gas chromatography method using a column having a length of 30 m or more, and a mass analyzer was installed on a detector of the separated molecules. The structure of the product identified by the above method is 7,10-epoxy octadeca-7,9-dienoic acid (EODA) on the 18 fatty acid chain The 7th carbon and the 10th carbon are connected by an epoxy structure and form a furan ring composed of 7th, 8th, 9th and 10th carbon. The results are shown in Fig.

Experimental Example 2: Bacterial bacteria and culture conditions

Clinical separated by deposited with the National pathogens Resource Bank as multidrug-resistant Staphylococcus aureus (Staphylococcus aureus) strain MDRSA 01ST001 the MRSA strains of (multidrug-resistant Staphylococcus aureus, MDRSA ) in 02ST085, 01ST093, 02ST165 and 02ST136 is Gyeongsangbuk University Hospital Was provided by the Pathogen Resource Bank. The strain was tested for the presence of pbp2a by oxoid latex agglutination assay. Methicillin-sensitive S. aureus (ATCC 29213) and Escherichia coli (ATCC 8739) were obtained from ATCC (American Type Culture Collection, Baltimore, USA). S. aureus 1199 (clinical isolates) was obtained from Wayne State University (MI, USA) and Salmonella typhimurium (KCTC 2515) was obtained from KCTC (Korean Collection for Type Cultures). All bacterial strains were cultured in TSB (trypticase soy broth) or NB (nutrient broth) medium purchased from Becton Dickinson (Cockeysville, MD, USA) and stored at -80 ° C in NB medium containing 25% glycerol. The cells were incubated overnight at the optimum temperature and used for the experiment.

Experimental Example 3: Reagents and substances

Antibiotics, chemicals and solvents used in the present invention were purchased from Sigma Aldrich (St. Louis, Mo., USA). All chemicals and reagents were analyzed using analytical grade. Furan fatty acid 7,10-EODA was prepared by using 7,10-EODA prepared in Experimental Example 1 and having a purity of 95% or more by gas chromatography.

Experimental Example 4. Measurement of minimal inhibitory concentration (MIC)

The minimum inhibitory concentration (MIC) of antibiotics and the activity of the 7,10-EODA of the present invention against Staphylococcus aureus strains were determined according to the CLSI guideline (CLSI document M100-517, 2007). Specifically, each antibiotic or 7,10-EODA was diluted 2-fold (200 - 0.97 μg / mL) by adding MHB (Mueller-Hinton Broth) to a 96-well microtiter plate. Culture in the mid-exponential phase (0.5 OD ₆₀₀ ) was diluted 10-fold with MHB and 0.1 mL aliquots of the diluent were added to the wells containing the serially diluted test reagent and incubated at 37 ° C for 12 hours Lt; / RTI > About the end of the incubation period, the cell density was measured at an optical density of 600 nm (OD ₆₀₀ ) using a culture medium. The minimum concentration required to prevent proliferation (greater than 90% relative to the control) was identified as MIC.

EXPERIMENTAL EXAMPLE 5 Rising by Agar Dispersion Analysis synergy effect screening

For synergistic screening, a 0.1 mL aliquot of MRSA or MSSA strain (5 x 10 ⁵ CFU / mL) cultured in a log-phase on TSA plates was inoculated. Then, two sets of sterile filter paper disks were placed on an agar plate, one set containing the antibiotic (1/4 x MIC) and the other set containing the 7,10-EODA (1/4 x MIC ) And enantiophene were impregnated together. DMSO alone or 7,10-EODA alone (1/4 x MIC) was used as a control. Plates were incubated at 27 ° C for 24 hours, and the synergistic effect was confirmed by ZOI (zone of inhibition).

Experimental Example 6 Rise (synergy)

Interactions of the selected antibiotic or 7,10-EODA of the present invention were evaluated using the checker board method (Nair et al. 2015). A 2-fold dilution of the serially diluted 7,10-EODA from 250 μg / mL (2 × MIC) stock was prepared in a 96-well microtiter plate column. To obtain various concentrations of 7,10-EODA and antibiotic combination, a 2-fold dilution of the beta-lactam antibiotic was placed on the same plate. About 10 ⁵ CFU / mL Microorganisms to be tested were added to the wells of the plates, and cultured for 24 hours at 37 ° C without shaking. At the end of the incubation period, cell growth was measured by OD ₆₀₀ using a spectrophotometer. Each 7,10-EODA and antibiotic combination was repeated at least 3 times. MIC was identified as the lowest concentration of antibiotics with over 90% growth inhibition. FIC _AB means the fractional inhibitory concentration (FIC) of drug A in the presence of drug B, and FIC _BA means the partial inhibitory concentration (FIC) of drug B in the presence of drug A. The fractional inhibitory concentration index (FICI) represents the sum of the FIC values of the interacting antibiotics. The interactions are based on FICI values, with FICIs less than 0.5 being categorized as synergy, FICIs greater than 0.5-4 being indifferent, and FICIs greater than 4 being antagonistic. In addition, the degree of elevation was measured based on the Bliss independence model of synergy (Morones-Ramirez et al. 2013). Specifically, the degree of elevation was calculated by the following equation: S is the degree of synergy, f _AB is the bacterial growth (optical density) when 7,10-EODA and antibiotics are combined, f _AO and f _0B represent bacterial growth for antibiotic alone or 7,10-EODA alone, respectively. f ₀₀ indicates bacterial growth in the absence of drug. The degree of elevation is divided into three categories based on the S value, 0 means neutral, 0 means synergism, 0 means antagonism.

Experimental Example  7. Time-kill assay

Increased interrelationships between 7,10-EODA and antibiotics were measured by changes in viability in time-kill analysis. A glass tube containing 1 mL of MHB and containing antibiotics alone (1/4 × MIC), 7,10-EODA alone (1/4 × MIC) or 7,10-EODA and antibiotics (About 10 ⁵ CFU / mL). MHB only cultures were used as controls. And incubated at 37 DEG C for 24 hours in a shaking incubator (150 rpm). At 0, 4, 8, and 24 hours, 0.1-mL aliquots were removed from the culture, and the cells were continuously diluted and cultured at 37 ° C for 24 hours on a plate containing MHA (Mueller-Hinton Agar). Survival colonies for each treatment group were counted and expressed as log CFU / mL relative to incubation time. Each treatment group was repeated three times and expressed as mean ± SD. A lower number of surviving colonies than 2.5 log CFUs was considered as an upward interaction (Lorian, 2005, Lippincott Williams & Wilkins, Philadelphia).

Example  One. Multidrug resistance  Staphylococcus aureus ( multidrug -Resistant Staphylococcus aureus, MDRSA), the antimicrobial activity of 7,10-EODA

The effect of the 7,10-EODA of the present invention on the multidrug-resistant Staphylococcus aureus (MDRSA) strain 01ST001 was confirmed.

Specifically, in order to compare the antimicrobial activity of the 7,10-EODA or the antibiotic of the present invention against the MDRSA 01ST001 strain, according to the method shown in Experimental Example 5, the 7,10-EODA (E 100 μg), streptomycin (S, 100 μg), ampicillin (100 μg), oxacillin (O, 100 μg), vancomycin (V, 100 μg) or penicillin , P, 100 [mu] g).

In order to confirm the antimicrobial activity according to the concentration of 7,10-EODA of the present invention, the concentrations of 7,10-EODA were respectively 50 μg (E1), 100 μg (E2), 200 μg (E3) (E4) and further treated with oxacillin (O, 100 g) or vancomycin (V, 100 g).

In order to confirm the effect of the combination treatment of the beta-lactam antibiotic against the MDRSA strain 01ST001 and the 7,10-EODA of the present invention, the beta-lactam antibiotic oxacillin (5 μg) or ampicillin (5 μg) + 7,10 -EODA (31.2 μg) treated group (A); (B) treated with oxacillin (100 μg) or ampicillin (100 μg) + 7,10-EODA (31.2 μg); Oxacillin (5 μg) or ampicillin (5 μg) alone group (a); Oxacillin (100 μg) or ampicillin (100 μg) alone group (b); And 7,10-EODA (31.2 μg) alone group (S). The results are shown in Fig.

As shown in Fig. 2 (1), it was confirmed that the MDRSA 01ST001 strain showed resistance to streptomycin (S), ampicillin (A), oxacillin (O) and penicillin (P) -EODA (E) showed antimicrobial activity against MDRSA 01ST001 strain.

As shown in FIG. 2 (2), it was confirmed that the antimicrobial activity against the MDRSA 01ST001 strain was high in a concentration-dependent manner of the 7,10-EODA of the present invention.

As shown in FIG. 2 (3), when oxacillin and the 7,10-EODA of the present invention were used as antibiotics, antimicrobial activity against the MDRSA 01ST001 strain compared to oxacillin alone group and 7,10- And the activity was greatly increased.

As shown in Fig. 2 (4), when the ampicillin and the 7,10-EODA of the present invention were used as the antibiotic, the antimicrobial activity against the MDRSA 01ST001 strain was higher than that of the ampicillin alone group and the 7,10-EODA alone group Respectively.

Example  2. MDRSA  The 7,10- EODA  And synergistic effect of antimicrobial activity by combination treatment of penicillin

The synergistic effect of 7,10-EODA and penicillin on the antimicrobial activity of multi-drug resistant Staphylococcus aureus (MDRSA) was confirmed.

Specifically, the method shown in Experimental Example 4 was used, and the concentration of penicillin was gradually reduced from 0 to a maximum concentration of 350 (μg / mL) by one-half at a maximum concentration. , 10-EODA at 0, 7.5, and 15.0 μg / mL, respectively, and cell growth of MDRSA 01ST001 strain was measured at an optical density of 600 nm (OD ₆₀₀ ). The results are shown in Fig.

As shown in FIG. 3, even when penicillin was treated at the same concentration, it was confirmed that when the concentration of 7,10-EODA to be used in combination increases, the growth of MDRSA 01ST001 strain is effectively inhibited. Specifically, MIC ₉₀ inhibits growth of 90% or more of the initial cell concentration When the value of 7,10-EODA was not co-administered, penicillin MIC ₉₀ The value was 350 μg / mL. On the other hand, the original MIC ₉₀ of 7,10-EODA When the concentration was 7.5 μg / mL corresponding to 1/16 of the value, the penicillin MIC ₉₀ The value decreased to 87.5 μg / mL, and the original MIC ₉₀ of 7,10-EODA When the concentration of 15.0 μg / mL corresponding to 1/8 of the value was used, penicillin MIC ₉₀ And the value decreased to 10.9 μg / mL.

Therefore, the 7,10-EODA of the present invention was originally classified as MIC ₉₀ The values of MIC _{90 of} penicillin treated in combination with 1/8, 1/16, The decrease in the value was found to be reduced to 1/4 and 1/32, respectively, as compared with the case where 7,10-EODA was not used in combination. Therefore, it was confirmed that the antibiotic activity synergistic effect by the combination treatment of 7,10-EODA of the present invention appears.

Example  3. Multidrug resistance  Staphylococcus aureus ( multidrug -resistant Staphylococcus aureus, MDRSA ) To 7,10- EODA  And the antibiotic activity synergistic effect by the combination treatment of each antibiotic

For the multidrug-resistant Staphylococcus aureus (MDRSA) strain (MDRSA 01ST001) or the non-drug-resistant Staphylococcus aureus (ATCC 29213) strain, the antibiotic treatment with 7,10-EODA And the antimicrobial activity synergistic effect was confirmed.

Specifically, the minimal inhibitory concentration (MIC) was measured according to the method of Experimental Example 4, and the degree of inhibition (ZOI) was measured according to the method of Experimental Example 5. Antibiotics include streptomycin (STP), ciprofloxacin (CIP), chloramphenicol (CPL), chlorpromazine (CPZ), rifampicin, rifampicin, irgasan, (TMP, trimethoprim), oxacillin (OXA, oxacillin), ampicillin (AMP, cephalexin) or penicillin (PEN, penicillin) were respectively used as 1/2 × MIC concentration. (MDRSA 01ST001) or non-MDRSA strain (MDRSA 01ST001) by treating or not treating the antibiotic of the present invention with a concentration of 31.2 μg of 7,10-EODA (1/4 × MIC) The synergistic effect of the combined treatment with the drug-resistant Staphylococcus aureus (ATCC 29213) strain was confirmed. The results are shown in Table 1 below.

Antibiotic MIC
(μg / mL) ZOI (mm)
without / with 7,10-EODA 001 29213 001 29213 STP 62.5 3.0 6.0 ± 0.0 / 12.9 ± 0.2 6.0 ± 0.0 / 11.4 ± 0.7 CIP 6.0 2.5 6.0 ± 0.0 / 13.2 ± 0.1 9.6 ± 0.2 / 13.2 ± 0.2 CPL 10.0 10.0 13.2 ± 0.4 / 13.9 ± 0.4 7.4 ± 0.2 / 7.21 ± 0.2 CPZ 62.5 125.0 7.2 ± 0.1 / 12.0 ± 0.5 6.0 ± 0.0 / 8.05 ± 0.5 RIF 5.0 5.0 18.3 ± 0.2 / 20.2 ± 0.1 21.0 ± 0.8 / 23.7 ± 1.2 IRG 0.1 0.1 12.4 ± 0.3 / 13.8 ± 1.1 11.1 ± 0.5 / 10.0 ± 0.3 TMP 7.8 7.8 6.0 0.0 / 6.00 0.0 8.9 ± 0.1 / 12.3 ± 0.2 OXA 62.5 0.04 6.0 ± 0.1 / 14.1 ± 0.2 8.3 ± 0.1 / 9.60 ± 0.4 AMP 1000.0 1.97 7.3 ± 0.2 / 14.2 ± 0.6 16.9 ± 0.3 / 17.0 ± 0.5 CEP 1000.0 1.97 10.1 ± 1.3 / 13.1 ± 0.6 15.3 ± 0.1 / 15.3 ± 0.2 PEN 1000.0 0.5 7.1 ± 0.1 / 13.6 ± 0.5 12.9 ± 0.4 / 13.2 ± 0.4

As shown in Table 1, when the combination of streptomycin and ciprofloxacin with 7,10-EODA of the present invention was used in combination with oxacillin, ampicillin and penicillin, which are beta-lactam antibiotics, the strain of multidrug-resistant Staphylococcus aureus MDRSA 01ST001) showed strong antimicrobial activity. In addition, when the antibiotic and the 7,10-EODA of the present invention were used in combination, it was confirmed that the ZOI (mm) value was increased to about 92-135% as compared with the antibiotic alone treatment group, Which was significantly increased.

Example  4. Multidrug resistance  Staphylococcus aureus ( multidrug -resistant Staphylococcus aureus, MDRSA ) Strains and Staphylococcus aureus strains, 7,10- EODA  And the antibiotic activity synergistic effect by the combination treatment of each antibiotic

(MDRSA 01ST001) which is a strain of multidrug-resistant Staphylococcus aureus (MDRSA), methicillin-resistant Staphylococcus aureus (MRSA) strains 085, 093, 120, 136 and 165, E. coli ATCC 8739 and S. typhimurium KCTC 2515, the synergistic effect of 7,10-EODA and each antibiotic was investigated. (A) 7,10-EODA alone group (E), oxacillin and 7,10-EODA combination treatment group (O + E (E)) in combination with oxacillin alone group ) And ampicillin and 7,10-EODA (A + E), and 7,10-EODA was treated at a concentration of 31.2 μg / mL (1/4 × MIC). Fractional inhibitory concentration indices (FICI) were confirmed. The results are shown in Table 2 below.

Strain MIC (μg / mL) MIC in combination (μg / mL) FICI O A E O + E A + E O + E A + E 085 250.0 1000.0 125.0 7.81 62.5 0.15 0.31 093 2000.0 31.2 125.0 125 3.90 0.31 0.37 165 31.2 2000.0 125.0 3.90 7.81 0.37 0.25 120 250.0 250.0 125.0 31.2 7.81 0.37 0.28 136 62.5 1000.0 125.0 3.90 15.6 0.31 0.26 001 62.5 1000.0 125.0 3.90 62.5 0.37 0.31 8739 200.0 0.6 500.0 200 0.6 2 2 2515 250.0 0.6 500.0 250 0.6 2 2

As shown in Table 2, when the 7,10-EODA of the present invention was used in combination with oxacillin, the FICI value was in the range of 0.25-0.37 and 0.299 on the average, indicating that the antimicrobial activity synergistic effect Whereas E. coli ATCC 8739 and S. typhimurium KCTC 2515, which are Gram-negative bacteria, showed an FICI value of 2.0, confirming the antimicrobial activity.

Example  5. Culture time and 7,10- EODA and  Antibiotic combination treatment MDRSA  01ST001 Confirmation of the growth of the strain

Each incubation time condition; And the growth of MDRSA 01ST001 strain according to the treatment conditions of 7,10-EODA and antibiotics.

Specifically, according to the method of Experimental Example 7, a 7,10-EODA treated group (a) of 15.6 μg concentration (1/8 × MIC) and a 7,10-EODA Treated group (a), oxacillin (c) at a concentration of 3.9 μg (c) and oxycillin treated group (a + c) at a concentration of 15.6 μg of 7,10-EODA plus 3.9 μg, and 7,10-EODA treated at a concentration of 31.2 μg (B + c) group treated with oxacillin at a concentration of 3.9 μg. Also, the growth of MDRSA 01ST001 strain was confirmed by cell density at 5, 10, 15, 20, 25 and 30 hours. The results are shown in Fig.

As shown in FIG. 4, the cell density of the strain MDRSA 01ST001 rapidly increased to 8.7 log CFU / mL in the control group in which nothing was treated. In addition, it was confirmed that the cell density was reduced to 1.30 log CFU / mL in the oxacillin treated group (a + c) of 7,10-EODA + 3.9 μg concentration of 15.6 μg concentration for 24 hours under the culture condition. In addition, the cell density was rapidly decreased to 2.5 log CFU / mL in the 7,10-EODA treated group and the 3.9 μg oxacillin treated group (b + c) at a concentration of 31.2 μg for 8 hours, And cell density of 3.9 log CFU / mL was observed. Therefore, it was confirmed that the combination of 7,10-EODA of the present invention and oxacillin as an antibiotic effectively inhibited the growth of MDRSA 01ST001 strain having multidrug resistance over time.

Example  6. 7,10- EODA  And non-beta-lactam antibiotic (non-β- lactam ) Combination of  Identification of antimicrobial activity synergistic effect on MRSA, MRSA or MSSA strains by treatment

The synergistic effect of antimicrobial activity against MRSA, MRSA or MSSA strains following combination treatment of 7,10-EODA and non-beta-lactam antibiotic (non-β-lactam) was confirmed.

Specifically, in accordance with the method shown in Experimental Example 6, the microorganism-resistant Staphylococcus aureus (MSSA) strain ATCC 29213, the multidrug-resistant Staphylococcus aureus (MDRSA) 01ST001 strain, the MRSA (methicillin-resistant Staphylococcus aureus) 01ST018, S. aureus strain 1199 was used. Each of the strains was incubated for 24 hours at a concentration of 15.6 μg / mL (1/8 × MIC) in combination with 7,10-EODA and antibiotics (1/4 × MIC). These antibiotics include rifampicin (RFC), chlorpromazine (CPZ), tetracycline (TET), gentamicin (GEN) and bacitracin (BAC) and vancomycin Respectively. The results are shown in Fig.

As shown in FIG. 5, when the 7,10-EODA and the gentamycin of the present invention were used in combination, a high synergy index of 1.32 or 1.37 was observed for the strain MRSA 018 or the strain MDRSA 01ST001, respectively. In addition, when the 7,10-EODA of the present invention and bacitracin or rifampicin were used in combination with the MDRSA 01ST001 strain, it was confirmed that the synergy index was 0.8 and 0.6, respectively.

Thus, through the above series of results, the 7,10-EODA of the present invention can be used in combination with various antibiotics. It was confirmed that the antibiotic synergistic effect against Staphylococcus aureus of various tolerance characteristics was shown without antagonism. It was also confirmed that the 7,10-EODA of the present invention is excellent in antimicrobial activity and antibacterial activity synergistic effect in combination with various antibiotics besides beta-lactam antibiotics.

Claims (30)

7,10-epoxy-octadeca-7,9-dienoic acid and penicillin antibiotics, aminoglycoside antibiotics, rifampicin antibiotics, polypeptide antibiotics, Antibiotics selected from the group consisting of fluoroquinolone antibiotics; As an active ingredient, an antibiotic-resistant microorganism. The method according to claim 1,
The 7,10-epoxyoctadeca-7,9-dienoic acid was prepared by heating 7,10-dihydroxy-8 (E) -octadecenoic acid with 7,10-dihydroxy- Wherein the antimicrobial composition is obtained by a method comprising the steps of: delete delete The method according to claim 1,
The penicillin antibiotics may be selected from the group consisting of benzylpenicillin, penoxymethylpenicillin, ampicillin, amoxycillin, bacampicillin, methicillin, oxacillin, An antibiotic-resistant antimicrobial composition for inhibiting antibiotics, which is characterized by being at least one antibiotic selected from the group consisting of Oxacillin, Cloxacillin, Piperacillin, Carbenicillin and Ticarcillin . delete delete delete delete The method according to claim 1,
The aminoglycoside antibiotic may be selected from the group consisting of amikacin, sisomycin, arbekacin, gentamycin, kanamycin, neomycin, spectinomycin, Wherein the antimicrobial composition is at least one antibiotic selected from the group consisting of spectinomycin, netilmicin, paromomycin, streptomycin and tobramycin. . delete delete delete The method according to claim 1,
The antimicrobial composition for inhibiting antibiotic-resistant bacteria, wherein the rifampicin-based antibiotic is rifamycin, rifampicin, or rifaximin. delete The method according to claim 1,
The antimicrobial composition for inhibiting antibiotic-resistant bacteria, wherein the polypeptide-based antibiotic is Polymyxin B or Bacitracin. delete The method according to claim 1,
The fluoroquinolone antibiotic is selected from the group consisting of Norfloxacin, Ciprofloxacin, Enrofloxacin, Ofloxacin and pefloxacin. An antimicrobial composition for inhibiting antibiotic-resistant bacteria, which is characterized by being at least one antibiotic. delete The method according to claim 1,
The antibiotic-resistant bacteria may be selected from the group consisting of Staphylococcus aureus , Escherichia coli , Salmonella enteritidis , Streptococcus pyogenes , Pseudomonas aeruginosa, wherein the antibiotic composition is derived from at least one bacterium selected from the group consisting of aeruginosa and Clostridium difficile . 21. The method of claim 20,
The Staphylococcus aureus may be selected from the group consisting of multidrug-resistant Staphylococcus aureus (MDRSA), methicillin-resistant staphylococcus aureus (MRSA), vancomycin resistant staphylococcus aureus , VRSA), and vancomycin intermediate staphylococcus aureus (VISA). The antimicrobial composition according to claim 1, The method according to claim 1,
The antimicrobial composition for inhibiting antibiotic-resistant bacteria, wherein the 7,10-epoxyoctadeca-7,9-dienoic acid is contained in the antimicrobial composition at a concentration of 0.01 to 5,000 g / ml. delete 7,10-epoxyoctadeca-7,9-dienoic acid; And antibiotics selected from the group consisting of penicillin antibiotics, aminoglycoside antibiotics, rifampicin antibiotics, polypeptide antibiotics and fluroquinolone antibiotics; As an active ingredient, a pharmaceutical composition for preventing or treating an infectious disease of an antibiotic-resistant microorganism. 25. The method of claim 24,
The antibiotic-resistant microorganism infectious disease is any one or more selected from the group consisting of abscesses, dermatitis, osteoarthritis, bacteremia, pneumonia, toxic shock syndrome, food poisoning, high fever, sepsis, edema, mastitis and intestinal colicosis. A pharmaceutical composition for preventing or treating a disease. 7,10-epoxyoctadeca-7,9-dienoic acid; And antibiotics selected from the group consisting of penicillin antibiotics, aminoglycoside antibiotics, rifampicin antibiotics, polypeptide antibiotics and fluroquinolone antibiotics; As an active ingredient, to an antibiotic resistant bacterium. 7,10-epoxyoctadeca-7,9-dienoic acid; And antibiotics selected from the group consisting of penicillin antibiotics, aminoglycoside antibiotics, rifampicin antibiotics, polypeptide antibiotics and fluroquinolone antibiotics; As an active ingredient, an antibiotic resistant bacterium. 7,10-epoxyoctadeca-7,9-dienoic acid; And antibiotics selected from the group consisting of penicillin antibiotics, aminoglycoside antibiotics, rifampicin antibiotics, polypeptide antibiotics and fluroquinolone antibiotics; As an active ingredient, to an antibiotic resistant bacterium. 7,10-epoxyoctadeca-7,9-dienoic acid; And antibiotics selected from the group consisting of penicillin antibiotics, aminoglycoside antibiotics, rifampicin antibiotics, polypeptide antibiotics and fluroquinolone antibiotics; As an active ingredient. An antimicrobial auxiliary for the killing of an antibiotic-resistant bacterium containing 7,10-epoxyoctadeca-7,9-dienoic acid as an active ingredient.

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