KR101671325B1 - Novel cyclic depsipeptide-based compound, separation method thereof, and antibacterial pharmaceutical composition containing the same as an active ingredient - Google Patents

Abstract

The present invention relates to a novel cyclic depsipeptide compound, a process for preparing the same, and a pharmaceutical composition for antibacterial activity containing the compound as an active ingredient. The novel cyclic depsipeptide compound, its optical isomer, The pharmaceutically acceptable salts exhibit inhibitory activity against Staphylococcus aureus and Salmonella typhimurium, and thus are useful as antimicrobial compositions, pharmaceutical compositions for antimicrobial use, antimicrobial feed compositions or antimicrobial cosmetic compositions Can be used.

Description

TECHNICAL FIELD The present invention relates to a novel cyclic depsipeptide compound, a process for preparing the same, and a pharmaceutical composition containing the cyclic depsipeptide-based compound as an active ingredient,

The present invention relates to a novel cyclic depsipeptide compound, a process for preparing the same, and a pharmaceutical composition for antibacterial activity containing the compound as an active ingredient.

The classical meaning of antibiotics is secondary metabolites produced by microorganisms, which means materials that kill or inhibit microbial growth at very low concentrations. The term antibiotics was first introduced by Waksman in the 1940s and was initially used to synthesize the synthetic sulfonamides and penicillins produced by the penicillium sp. It was used to distinguish. It has been difficult to uniquely define the meaning of antibiotics as a result of the discovery and synthesis of many antimicrobial activity substances. Currently, it is generally accepted that antibiotics are natural, semi- Is taken to mean a synthetic compound.

Researches on the search and use of natural products have explored and utilized the biosynthesis ability of organisms, and so far, mainly plants and microorganisms have been the main species of the subject. In particular, microorganisms have a variety of species and produce unique and interesting physiologically active substances, have a short generation period, are easy to mass-produce, and have high industrial availability, and thus have become attractive sources of physiologically active substances. The search for useful substances produced by microorganisms has been actively conducted mainly on land microorganisms, and many of them have been established in usefulness in fermentation industry and medicine. In addition, recently, antibiotic-resistant bacteria appearing resistant to conventional antibiotics are emerging and new diseases such as acquired immunodeficiency syndrome (AIDS) are increasing and new drugs that can replace conventional antibiotics are urgently needed .

In this connection, Patent Document 1 (Korean Patent Laid-Open Publication No. 10-2007-0086038) discloses a compound having a depressed peptide parent nucleus and a composition for an antimicrobial composition containing the same. In addition, Patent Document 2 (Korean Patent Laid-Open Publication No. 10-1986-0000382) discloses a compound having a depsipeptide nucleotide and an antitumor or antibiotic composition containing the same.

Previously developed antibiotics are classified as beta-lactam class of antibiotics. There are methicillin, which is considered to be superior to penicillin, and is resistant to methicillin And vancomycin, which has been developed to have an antibacterial effect against bacteria, specifically methicillin resistance staphylococcus aureus (MRSA).

However, in recent years, resistance to existing antibiotics such as vancomycin resistance staphylococcus aureus (VRSA), which is called superbacteria, has increased, and the necessity of development of a substance or drug having resistance to antibiotics It is being stressed. In other words, there is a growing interest in the development of drugs with new mechanisms of action that can replace existing antibiotics. Therefore, it is required to develop a target specific search method and new resources for the development of new active materials.

Accordingly, the present inventors have studied a compound having excellent inhibitory activity against a specific bacterium, and found that a novel cyclic depsipeptide compound according to the present invention, an optical isomer thereof, or a pharmaceutically acceptable salt thereof is useful as a staphylococcus aureus and Salmonella typhimurium, and thus it is useful as a composition for antimicrobial use, a pharmaceutical composition for antimicrobial use, an antimicrobial feed composition or an antimicrobial cosmetic composition, and completed the present invention.

It is an object of the present invention to provide a compound having an inhibitory activity against Staphylococcus aureus and Salmonella typhimurium, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a process for preparing the above compound, its optical isomer, or a pharmaceutically acceptable salt thereof.

Still another object of the present invention is to provide an antimicrobial composition containing the above compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for antimicrobial use containing the compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Yet another object of the present invention is to provide an antimicrobial feed composition containing the above compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide an antimicrobial cosmetic composition comprising the compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Yet another object of the present invention is to provide a strain of Streptomyces sp. KCB13F003 which produces the above compound.

In order to achieve the above object, the present invention provides a compound represented by the following general formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

In Formula 1,

R ¹ is -H, -OH, halogen, nitrile, C _{1 -10} linear or branched alkyl, or C _{1 -10} linear or branched alkoxy.

The present invention also relates to a method for producing a strain of Streptomyces sp. KCB13F003 (step 1);

Extracting the strain culture obtained in step 1 to obtain an extract (step 2); And

And a step of purifying the extract obtained in the step 2 (step 3).

Furthermore, the present invention provides an antimicrobial composition comprising the compound represented by the above-mentioned formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention also provides a pharmaceutical composition for antimicrobial use comprising the compound represented by the above-mentioned formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Furthermore, the present invention provides an antimicrobial feed composition comprising the compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention also provides an antimicrobial cosmetic composition comprising the compound represented by the formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Further, the present invention provides a Streptomyces sp. KCB13F003 strain producing the above compound.

The novel cyclic depsipeptide compound according to the present invention, an optical isomer thereof, or a pharmaceutically acceptable salt thereof exhibits an inhibitory activity against Staphylococcus aureus and Salmonella typhimurium, A pharmaceutical composition for antimicrobial use, an antimicrobial feed composition or an antimicrobial cosmetic composition.

1 is an image showing the ¹ H NMR spectrum (800 MHz, DMSO-d ₆ ) of the compound represented by the formula (A) prepared in Example 4;
2 is an image showing the ¹³ C NMR spectrum (200 MHz, DMSO-d ₆ ) of the compound represented by the formula (A) prepared in Example 4;
3 is an image showing the ¹ H NMR spectrum (700 MHz, DMSO-d ₆ ) of the compound represented by the formula (B) prepared in Example 4; And
4 is an image showing the ¹³ C NMR spectrum (225 MHz, DMSO-d ₆ ) of the compound represented by the formula (B) prepared in Example 4. Fig.

Hereinafter, the present invention will be described in detail.

The present invention provides a compound represented by the following general formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

In Formula 1,

R ¹ is -H, -OH, halogen, nitrile, C _{1 -10} linear or branched alkyl, or C _{1 -10} linear or branched alkoxy.

Preferably,

R ¹ is -H, -OH, halogen, C _{1 -5} straight or branched chain alkyl, or C _{1 -5} straight or branched alkoxy.

More preferably,

R ¹ is -H or -OH.

The compound represented by the formula (1) of the present invention can be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include those derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid and the like, aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, Derived from organic acids such as acetic acid, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid and the like. Examples of such pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate chloride, bromide, But are not limited to, but are not limited to, but are not limited to, but are not limited to, but are not limited to, halides, halides, halides, halides, halides, halides, But are not limited to, lactose, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, Methoxybenzoate, phthalate, terephthalate, benzene sulfonate, toluene sulfonate, chlorobenzene Sulfonates, methanesulfonates, propanesulfonates, naphthalene-1-sulfonates, and the like, as well as sulfonates such as benzyl sulfonate, sulfonate, xylene sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, -Sulfonate, naphthalene-2-sulfonate, mandelate, and the like.

The acid addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving a derivative of the formula (1) in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile and the like, Followed by filtration and drying. Alternatively, the solvent and excess acid may be distilled off under reduced pressure, followed by drying and crystallization in an organic solvent.

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

Furthermore, the present invention encompasses the compounds represented by the formula (1) and pharmaceutically acceptable salts thereof as well as solvates, optical isomers and hydrates thereof which can be prepared therefrom.

The novel cyclic depsipeptide compound of the present invention is a compound prepared from a strain of ground soil actinomycetes, preferably Streptomyces sp., More preferably Streptomyces sp. KCB13F003 strain. Is characterized by having a chemical structure unknown to the polymer.

The present invention also relates to a method for producing a strain of Streptomyces sp. KCB13F003 (step 1);

Extracting the strain culture obtained in step 1 to obtain an extract (step 2); And

And a step of purifying the extract obtained in the step 2 (step 3).

Hereinafter, the process for preparing the compound represented by the formula (1) according to the present invention will be described in detail.

In the method for producing a compound represented by the formula (1) according to the present invention, the step (1) is a step of culturing a strain of Streptomyces sp. KCB13F003 to obtain a strain culture. Specifically, the culture of the strain is cultured in a medium containing a nutrient source to which ordinary microorganisms can be used. As a nutrient source, a known nutrient source conventionally used for culturing actinomycetes is used. For example, the carbon source may be glucose, starch syrup, dextrin, starch, molasses, animal oil, vegetable oil, etc. The nitrogen sources may include wheat bran, soybean meal, wheat, malt, cottonseed oil, Extract, ammonium sulfate, sodium nitrate, urea, and the like. If necessary, it is very effective to add sodium chloride, potassium, magnesium, cobalt, chlorine, phosphoric acid, sulfuric acid and other inorganic salts promoting ion production. As the culture method, it is possible to cultivate shaking or stationary culture under aerobic conditions.

The culture temperature may vary slightly depending on conditions when culturing under the above conditions, but it is generally preferable to culture at 20 to 37 ° C, more preferably at 25 to 30 ° C.

In the method for producing the compound represented by the formula (1) according to the present invention, the step (2) is a step of extracting the culture obtained in the step (1) with a solvent to obtain an extract. In general, the cyclic depsipeptide-based compound is present not only in the culture medium of the strain but also in the cell portion. Therefore, a solvent is added to the culture medium and cells of the strain, and the effective component is extracted from the culture medium and the cells, and the obtained extract is concentrated by reduced-pressure evaporation.

At this time, ethyl acetate is preferably used as the solvent.

In the method for producing the compound represented by the formula (1) according to the present invention, the step (3) is a step for purifying the extract obtained in the step (2). Specifically, the extract obtained in step 2 is subjected to flash column chromatography using a methanol: water mixed solvent, and then purified by high performance liquid chromatography to prepare a compound represented by formula (1).

The present invention also provides an antimicrobial composition comprising the compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Here, the bacterium may be a Staphylococcus sp. Bacterium or a Salmonella sp. Bacterium, more preferably a Staphylococcus sp. Bacterium or a Salmonella sp. The bacteria may be, but are not limited to, Staphylococcus aureus or Salmonella typhimurium, respectively.

Further, the present invention provides a pharmaceutical composition for antimicrobial use containing the compound represented by the formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In this connection, as a result of conducting experiments to evaluate the antimicrobial activity of the compounds represented by the chemical formulas A and B according to the present invention, the compounds represented by the chemical formulas A and B according to the present invention were found to be Salmonella and Staphylococcus aureus (See Table 4 in Experimental Example 2). ≪ tb >< TABLE >

Therefore, the novel cyclic depsipeptide compound according to the present invention, its optical isomer, or a pharmaceutically acceptable salt thereof exhibits an inhibitory activity against Staphylococcus aureus and Salmonella typhimurium, A pharmaceutical composition for antimicrobial use, an antimicrobial feed composition or an antimicrobial cosmetic composition.

The compounds according to the present invention are generally administered in the form of a pharmaceutical composition comprising the active compound of the present invention and comprising a pharmaceutically acceptable carrier or carrier suitable for use in a pharmaceutical preparation.

Furthermore, the present invention provides an antimicrobial feed composition comprising the compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention also provides an antimicrobial cosmetic composition comprising the compound represented by the formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Furthermore, the present invention provides a strain of Streptomyces sp. KCB13F003 producing the compound represented by the above formula (1).

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples.

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

< Example  1> Streptomyces  genus( Streptomyces sp .) KCB13F003  Isolation of strain

This strain was isolated from the soil samples of Ulleungdo in August 2013. The collected soil samples were stored in a sterilized plastic bag immediately after collection and refrigerated until used for experiment. Soil samples were diluted 10 times with sterile distilled water. 1 ml of the diluted solution was plated on a Starch-Yeast extract agar medium and cultured at 28 ° C for 5 days to purify the colonies.

< Example  2> Identification and naming of strains

As a result of GenBack search for the nucleotide sequence obtained through rRNA sequencing of this strain, the strain showed 99.58% homology to Streptomyces sp. (SEQ. Thus, this strain was identified as Streptomyces sp. And named Streptomyces sp. KCB13F003 (accession number: KCTC12731BP).

< Example  3> Streptomyces  genus( Streptomyces sp .) KCB13F003  Culture of strain

In order to cultivate the actinomycetes strain, a medium containing nutrients conventionally used by microorganisms was prepared. (W / v) lactose, 0.5% (w / v) malt extract, 0.5% (w / v) yeast extract and 0.1% (w / v) lactose as the crude medium and production medium of actinomycetes / v) GLY medium containing calcium carbonate was used.

The 500 ml Erlenmeyer flask containing 100 ml of the medium described above was sterilized at 121 ° C for 15 minutes and platinum was inoculated from a slope culture test tube of Streptomyces sp. KCB13F003 strain, followed by shaking culture for 3 days, Respectively. 30 ml of a 1,000 ml Erlenmeyer flask containing 250 ml of production medium was inoculated with 3 ml of each seed culture, followed by shaking culture at 28 ° C for 5 days.

< Example  4> Ring type Depsipeptide system  Preparation of compounds

The culture of Streptomyces sp. KCB13F003 strain cultivated in Example 3 was extracted with ethyl acetate (15 L), and the obtained extract was concentrated by reduced pressure evaporation using a vacuum dryer. The concentrate was adsorbed on Odyssey 18 and subjected to ODS RP-18 flash column chromatography, in which methanol / water (8: 2-10 / 0, v / v) And eluted with increasing methanol concentration stepwise as a solvent. At this time, the fraction containing the compound represented by the following chemical formula A (designated as urenogagemide A) and the compound represented by chemical formula B (designated as urendogamide B) was eluted from 80% methanol. The fractions were concentrated under reduced pressure and then acetonitrile and water were eluted with a gradient of 40:60 to 80:20 using a high performance liquid chromatography (column: Optimapak C18, length 25 mm, diameter 10 mm) ml / min to prepare a compound represented by the following formula (A) wherein the UV absorption peak at 210 nm was observed at 17 minutes.

(A)

The same fraction was eluted with a gradient of acetonitrile and water at a flow rate of 3 ml / min with a gradient of 40: 60 - 52: 48 using the same column, and the UV absorption peak at 210 nm was shown in the following formula (B) &Lt; / RTI &gt;

[Chemical Formula B]

< Experimental Example  1> Ring type Depsipeptide system  Structural analysis of compounds

The molecular weight and the molecular formula of the compound represented by the formula (A) prepared from the culture solution of Streptomyces sp. KCB13F003 strain were determined using an ESIMS mass spectrometer (Electrospray Ionization mass spectrometer). Further, ¹ H NMR, ¹³ C NMR, COZY (Correlation Spectroscopy) and HMQC (Bruker Biospin Advance II 900 NMR spectrometer, Bruker AVANCE HD 800 NMR spectrometer, Bruker AVANCE HD 700 NMR spectrometer) Hetero-nucleated Multiple-Bond Coherence (HMBC), Distortionless Enhancement by Polarization (DEPT) and Nuclear Overhauser Effect Spectroscopy (NOESY) spectra were obtained and the molecular structure of the compounds was determined.

The measurement results are as follows, and the substance prepared from the culture solution of Streptomyces sp. KCB13F003 strain was identified as a novel cyclic depsipeptide compound represented by the above formula (A) and (B).

The compound represented by the formula (A) is a compound represented by the following formula (1): phenylalanine, glycine, threonine, N-methylphenylalanine, 2- isopropylsuccinic acid, 5-hydroxy- ( ¹ H, &lt; ¹³ &gt; C NMR data is shown in Table 1 below).

A compound represented by the formula (A): white powder; [?] _D +88.4 ( c 0.05, MeOH); UV (MeOH)? _Max (log?) 204 (3.6), 210 (3.3); HRESIMS m / z 986.4861 [M + H] + (calcd for C 51 H 68 N 7 O 13, 986.4875).

δ _C ^b , type 隆 _H ^c , mult ( J in Hz) δ _C ^b , type 隆 _H ^c , mult ( J in Hz) 5- Hydroxy -6- methyl -
2,3- Dehydropiperboxylic acid Glycine One 162.3, C 29 168.0, C 2 129.4, C 30 40.5, CH ₂ 4.54, m 3  120.2, CH 5.98, s 3.48, m 4 27.3, CH ₂
2.28, ovl ^a
2.01, ovl ^a 30-NH 8.04, d (7.5) Threonine 5 65.2, CH 3.75, s 31 167.4, C 6 53.6, CH 3.93 d (5.7) 32 54.6, CH 4.77, d (9.1) 7 14.5, CH ₃ 0.97, ovl ^a 33 71.4, CH 5.48, bd (3.1) 5-OH 4.79, ovl ^a 34 15.9, CH ₃ 0.97, ovl ^a Piperaconic acid 32-NH 7.69, d (8.1) 8 170.6, C N- methyl - phenylalanine 9 50.2, CH 5.56, ovl ^a 35 170.9, C 10 26.7, CH ₂ 1.94, m
1.59, ovl ^a 36 56.7, CH 5.57, ovl ^a 37 34.7, CH ₂ 3.20, dd (14.4, 5.4) 11 19.6, CH ₂ 1.53, ovl ^a
1.31, m 2.90, dd (13.8, 11.0) 38 137.8, C 12 24.7, CH ₂ 1.52, ovl ^a
1.24, m 39 129.4, CH 7.20, ovl ^a 40 128.8, CH 7.23, ovl ^a 13 42.7, CH ₂ 3.89, bd (13.3)
3.10, t (12.5) 41 126.1, CH 7.16, ovl ^a Phenylalanine 42 128.8, CH 7.23, ovl ^a 14 170.0, C 43 129.4, CH 7.20, ovl ^a 15 48.4, CH 5.15, dd (16.2, 8.4) 44 31.3, CH ₃ 2.94, s 16 37.6, CH ₂ 3.00, ovl ^a
2.82, dd (13.6, 8.5) 2- Isopropylsuccinic acid 45  172.0, C 17 137.6, C 46 31.8, CH ₂ 2.52, ovl ^a 18 129.4, CH 7.20, ovl ^a 2.10, d (14.9) 19 128.8, CH 7.23, ovl ^a 47 46.8, CH 2.48, ovl ^a 20 126.2 7.16, ovl ^a 48 29.2, CH 1.73, m 21 128.8 7.23, ovl ^a 49 19.7, CH ₃ 0.78, d (6.4) 22 129.4 7.20, ovl ^a 50 19.7, CH ₃ 0.76, d (6.4) 15-NH 8.90, d (9.4) 51 175.6, C 4- Hydroxypiperidine 23 170.8, C 24 52.4, CH 4.29, m 25 33.2, CH ₂ 1.82, ovl ^a
1.41, m 26 61.9, CH 3.65, bs 27 31.3, CH ₂ 1.59, ovl ^a
1.45, m 28 34.1, CH ₂ 4.17, m
3.19, ovl ^a 26-OH 4.43, bs

In Table 1,

- ^a represents ^a superimposed signal;

- ^b indicates measurement at 200 MHz; And

- ^c indicates measurement at 800 MHz.

The compound represented by the general formula (B) is the same as the compound represented by the general formula (A) in the structure of phenylalanine, glycine, threonine, N-methyl-phenylalanine, 2-isopropylsuccinic acid, Hydroxy-6-methyl-2,3-dehydropiperboxylic acid is converted to 4,5-dihydroxy-6-methyl-2,3-dehydropiperic acid, ( ¹ H, &lt; ¹³ &gt; C NMR data are shown in Table 2 below).

A compound represented by the formula (B): white powder; [[alpha]] _D +69.6 ( c 0.05, MeOH); UV (MeOH)? _Max (log?) 204 (3.6); HRESIMS m / z 1002.4818 [M + H] ⁺ (calcd for C ₅₁ H ₆₈ N ₇ O ₁₄ , 1002.4824).

δ _C ^b , type 隆 _H ^c , mult ( J in Hz) δ _C ^b , type 隆 _H ^c , mult ( J in Hz) 4,5-dihydroxy-6-methyl-
2,3-dehydropiphenic acid Glycine One 162.4, C 29 168.1, C 2 130.4, C 30 40.4, CH ₂ 4.56, dd (18.0, 7.4) 3  121.4, CH 5.73, s 3.43, ovl ^a 4 62.9, CH 4.16 30-NH 8.04, d (7.7) 5 66.9, CH 3.60, bs Threonine 6 56.0, CH 4.08, m 31 167.3, C 7 14.5, CH ₃ 1.09, d (6.6) 32 54.5, CH 4.77, dd (9.1, 2.3) 4-OH Not detected 33 71.7, CH 5.49, dd (6.0, 3.1) 5-OH 4.82, ovl ^a 34 15.7, CH ₃ 0.97, d (6.2) Piperaconic acid 32-NH 7.79, d (8.2) 8 170.9, C N- methyl - phenylalanine 9 50.2, CH 5.54, ovl ^a 35 170.9, C 10 26.9, CH ₂ 1.89, m
1.57, m 36 56.8, CH 5.53, ovl ^a 37 34.8, CH ₂ 3.22, dd (14.1, 5.7) 11 19.6, CH ₂ 1.54, ovl ^a
1.28, ovl ^a 2.89, dd (14.1, 10.5) 38  137.8, C 12 24.7, CH ₂ 1.50, ovl ^a
1.24, ovl ^a 39 129.3, CH 7.20, ovl ^a 40 128.8, CH 7.26, ovl ^a 13 42.6, CH ₂ 3.91, t (14.0)
3.08, t (12.2) 41 126.2, CH 7.16, ovl ^a Phenylalanine 42 128.8, CH 7.26, ovl ^a 14 169.8, C 43 129.3, CH 7.20, ovl ^a 15 48.4, CH 5.13, ovl ^a 44 31.5, CH ₃ 2.92, s 16 37.6, CH ₂ 3.00, ovl ^a
2.82, dd (13.6, 8.5) 2- Isopropylsuccinic acid 45  171.9, C 17  137.6, C 46 31.9, CH ₂ 2.53, ovl ^a 18  129.3, CH 7.20, ovl ^a 2.09, dd (16.1, 3.5) 19  128.8, CH 7.26, ovl ^a 47 46.8, CH 2.46, m 20  126.2, CH 7.16, ovl ^a 48 29.3, CH 1.73, m 21  128.8, CH 7.26, ovl ^a 49 19.7, CH ₃ 0.78, ovl ^a 22  129.3, CH 7.20, ovl ^a 50 19.7, CH ₃ 0.76, ovl ^a 15-NH 8.95, d (9.8) 51 175.6, C 4- Hydroxypiperidine 23  170.8, C 24   52.5, CH 4.27, m 25 33.3, CH ₂ 1.81, m
1.39, m 26   61.9, CH 3.65, bs 27 31.3, CH ₂ 1.60, m
1.43, m 28 34.3, CH ₂ 4.17, m
3.16, t (12.8) 26-OH 4.43, ovl ^a

In Table 2,

- ^a represents ^a superimposed signal;

- ^b represents the measurement at 200 MHz; And

- ^c 800 MHz. &Lt; / RTI &gt;

< Experimental Example  2> Antimicrobial activity evaluation

The strains obtained from the accumulation culture collection of the Korean Gene Bank (KCTC) were used for the evaluation of antimicrobial activity, which is shown in Table 3 below.

Strain badge Salmonella
(Salmonella typhimurium, KCTC 1926) NA Staphylococcus aureus
(Staphylococcus aureus, KCTC 1916) LB agar Enterococcus lacalis
(Enterococcus faecalis, KCTC 3206) Brain Heart Infusion Agar Bacillus Subtilis
(Bacillus subtilis, KCTC 1022) NA Escherichia coli
(Escherichia coli, KCTC 1039) Brain Heart Infusion Agar Microkocas rutheus
(Micrococcus luteus, IAM 1056) ENB Agar Candida Albikans
(Candida albicans, KCTC 7678) YM Agar Penicillium greece
(Penicillium griseofulvum, KCTC 6435) Malt Extract Agar

In Table 3,

- NA (Nutrient Agar): [0.3% (w / v) beef extract, 0.5% (w / v) peptone, 1.5% (w / v) agar);

- LB (Lysogeny broth) Agar: [1% (w / v) tryptone, 0.5% (w / v) yeast extract, 1% v) agar;

Brain Heart Infusion Agar: [3.7% (w / v) BHI medium (Difco 0037), 1.5% (w / v) agar];

- Enriched Nutrient Broth (ENB) Agar: [1.25% (w / v) HI medium (difom 0038), 0.54% (w / v) nutrient broth, Difco 0003, 0.25% v) yeast extract];

YM Agar: 0.3% (w / v) yeast extract, 0.3% (w / v) malt extract, 0.5% (w / v) peptone, 1% Trosose, 2% (w / v) agar]; And

- Malt Extract Agar: [2% (w / v) malt extract, 2% (w / v) glucose, 0.1% (w / v) peptone, 2% (w / v) agar].

The antimicrobial activity was evaluated by the paper disc method. The agar medium to which each test solution was added was poured into a petri dish and a plate medium was prepared. After 100 μg, 50 μg and 30 μg of the compound represented by the formula (A) and the compound (B) were absorbed into a sterilized paper disc , Placed on a prepared plate for activity evaluation, and incubated at 37 占 폚 (Salmonella, Staphylococcus aureus, Enterococcus faecalis, E. coli) and 28 占 폚 (Bacillus subtilis, Micrococus rutherus, Candida albicans, The clear zone (mm) around the disc was measured by incubating in a static incubator of Penicillium griseo-ovary for 18 hours. The results are shown in Table 4 below.

Strain Clear zone (mm) The compound of formula A
(/ / Disc) Compound (B)
(/ / Disc) 100 50 30 100 50 30 Salmonella
(Salmonella typhimurium, KCTC 1926) 12.7 9.8 - - - - Staphylococcus aureus
(Staphylococcus aureus, KCTC 1916) 17.6 13.7 10.8 13.0 - - Enterococcus lacalis
(Enterococcus faecalis, KCTC 3206) - - - - - - Bacillus Subtilis
(Bacillus subtilis, KCTC 1022) - - - - - - Escherichia coli
(Escherichia coli, KCTC 1039) - - - - - - Microkocas rutheus
(Micrococcus luteus, IAM 1056) - - - - - - Candida Albikans
(Candida albicans, KCTC 7678) - - - - - - Penicillium greece
(Penicillium griseofulvum, KCTC 6435) - - - - - -

In Table 4,

The symbol "- " indicates that there is no activity against the strain.

As shown in Table 4, the compounds represented by the formula (A) and the formula (B) according to the present invention have an antibiotic effect specific to Salmonella and Staphylococcus aureus.

Therefore, the novel cyclic depsipeptide compound according to the present invention, its optical isomer, or a pharmaceutically acceptable salt thereof exhibits an inhibitory activity against Staphylococcus aureus and Salmonella typhimurium, A pharmaceutical composition for antimicrobial use, an antimicrobial feed composition or an antimicrobial cosmetic composition.

Meanwhile, the compounds of the present invention can be formulated into various forms depending on the purpose. Examples of formulations for the composition of the present invention are illustrated below.

< Formulation example  1> Manufacture of flexible lotion

To the purified water was added a mixture prepared by adding butylene glycol, glycerin, polyoxyethylene (60) hardened castor oil, betaine, citric acid, sodium citrate and an antiseptic, stirring, dissolving and then adding the flavor to ethanol. The compound represented by the formula (1) according to the present invention was added thereto and sufficiently stirred, followed by aging to prepare a flexible lotion. The content of each component is shown in Table 5 below.

Raw material Content (w / w%) The compound represented by the formula (1) 50.0 Butylene glycol 7.0 glycerin 5.0 Polyoxyethylene (60) Hardened castor oil 0.2 ethanol 5.0 Betaine 2.0 Citric acid 0.02 Sodium citrate 0.06 antiseptic a very small amount Spices a very small amount Purified water a very small amount

< Formulation example  2> Manufacture of nutrition lotion

The compound represented by Chemical Formula 1 according to the present invention, butylene glycol, glycerin, carboxyvinyl polymer, arginine, preservative and purified water were heated at 70 to 75 캜 with stirring. After adding a mixture of squalane, butylene glycol dicaprylate / dicaprate, sorbitan stearate, polysorbate 60, glyceryl stearate and stearyl glycerate, heated at 75 to 80 ° C with stirring, emulsified, While stirring, the mixture was cooled to about 45 ° C, and the fragrance was added and stirred. Thereafter, the mixture was cooled to 30 占 폚 and aged to prepare a nutrition lotion. The content of each component is shown in Table 6 below.

Raw material Content (w / w%) The compound represented by the formula (1) 40.0 Butylene glycol 8.0 glycerin 5.0 Squalane 10.0 Butylene glycol dicaprylate / dicaprate 5.0 Sorbitan stearate 1.5 Polysorbate 60 1.0 Glyceryl stearate 0.5 Stearyl glycyrrhetinate 0.2 Carboxyvinyl polymer 0.1 Arginine 0.1 antiseptic a very small amount Spices a very small amount Purified water a very small amount

< Formulation example  3> Production of Essence

Polyglyceryl 2-olate, ceramide, cetearase-4 and cholesterol were mixed with stirring, and then cytociterol, polyglyceryl 2-oleate, ceramide, cetearase-4 and cholesterol were stirred The mixture was emulsified by adding a compound represented by the formula (1) of the present invention, dicetylphosphate, concentrated glycerin and purified water. Thereafter, when the mixture was cooled to 45 캜 with stirring, a flavoring agent was added, stirred, and cooled to 30 캜, followed by aging. Carboxyvinyl polymer, xanthan gum and a preservative were added to the mixture to stabilize it, followed by aging to prepare an essence. The content of each component is shown in Table 7 below.

Raw material Content (w / w%) The compound represented by the formula (1) 10.0 Cytostelo 1.7 Polyglyceryl 2-oleate 1.5 Ceramide 0.7 Setares-4 1.2 cholesterol 1.5 Dicetyl phosphate 0.4 Concentrated glycerin 0.5 Carboxyvinyl polymer 0.2 Xanthan gum 0.2 antiseptic a very small amount Spices a very small amount Purified water a very small amount

< Formulation example  4> Manufacture of ointment

Ointment was produced by a conventional method according to the formulation shown in Table 8 below.

Ingredients role Amount of blending
(weight%) vaseline 40.0 Stearyl alcohol 15.0 The 15.0 POE (10) oleate 0.25 Glyceryl monostearate 0.25 Allantoin Anti-inflammatory (whitening) 1.0 Sorbitol Moisturizing component 5.0 Propylene glycol 5.0 Gentiana extract * 1 Moisturizing component 0.3 The compound represented by the formula (1) 6.0 antiseptic Proper amount Ion-exchange water Balance

* 1: Gentiana extract BG (manufactured by Maruzen Pharmaceutical Co., Ltd.) was used as the Gentiana extract.

Korea Biotechnology Research Institute (KCTC) KCTC12731BP 20141209

<110> Korea Research Institute Bioscience and Biotechnology <120> Novel cyclic depsipeptide-based compound, separation method          thereof, and an antibacterial pharmaceutical composition.          the same as an active ingredient <130> 2014P-12-022 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 1422 <212> DNA <213> Streptomyces staurosporinius <400> 1 gcgtgcttaa cacatgcaag tcgaacgatg aagccgcttc ggtggtggat tagtggcgaa 60 cgggtgagta acacgtgggc aatctgccct tcactctggg acaagccctg gaaacggggt 120 ctaataccgg atacgacacg gggtcgcatg acctccgtgt ggaaagctcc ggcggtgaag 180 gatgagcccg cggcctatca gcttgttggt ggggtaatgg cctaccaagg cgacgacggg 240 tagccggcct gagagggcga ccggccacac tgggactgag acacggccca gactcctacg 300 ggaggcagca gtggggaata ttgcacaatg ggcgaaagcc tgatgcagcg acgccgcgtg 360 agggatgacg gccttcgggt tgtaaacctc tttcagcagg gaagaagcga gagtgacggt 420 acctgcagaa gaagcgccgg ctaactacgt gccagcagcc gcggtaatac gtagggcgca 480 agcgttgtcc ggaattattg ggcgtaaaga gctcgtaggc ggcttgtcac gtcggatgtg 540 aaagcccggg gcttaacccc gggtctgcat tcgatacggg caggctagag ttcggtaggg 600 gagatcggaa ttcctggtgt agcggtgaaa tgcgcagata tcaggaggaa caccggtggc 660 gaaggcggat ctctgggccg atactgacgc tgaggagcga aagcgtgggg agcgaacagg 720 attagatacc ctggtagtcc acgccgtaaa cgttgggaac taggtgtggg cgacattcca 780 cgtcgtccgt gccgcagcta acgcattaag ttccccgcct ggggagtacg gccgcaaggc 840 taaaactcaa aggaattgac gggggcccgc acaagcagcg gagcatgtgg cttaattcga 900 cgcaacgcga agaaccttac caaggcttga catacgccgg aaaaccctgg agacagggtc 960 ccccttgtgg tcggtgaca ggtggtgcat ggctgtcgtc agctcgtgtc gtgagatgtt 1020 gggttaagtc ccgcaacgag cgcaaccctt gttctgtgtt gccagcatgc ccttcggggt 1080 gatggggact cacaggagac tgccggggtc aactcggagg aaggtgggga cgacgtcaag 1140 tcatcatgcc ccttatgtct tgggctgcac acgtgctaca atggccggta caatgagctg 1200 cgataccgcg aggtggagcg aatctcaaaa agccggtctc agttcggatt ggggtctgca 1260 actcgacccc atgaagtcgg agttgctagt aatcgcagat cagcattgct gcggtgaata 1320 cgttcccggg ccttgtacac accgcccgtc acgtcacgaa agtcggtaac acccgaagcc 1380 ggtggcccaa ccccttgtgg gagggaatcg tcgaaggtgg ga 1422

Claims (10)

Claims 1. A compound represented by the following formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof:
[Chemical Formula 1]

(In the formula 1,
R ¹ is -H, -OH, a halogen, a nitrile, a linear or branched alkyl, or straight or branched chain alkoxy of C _{1 -10} of C _{1 -10).}
The method according to claim 1,
R ¹ is -H, -OH, halogen, C _{1 -5} straight or branched chain alkyl, or C _{1 -5} straight or branched alkoxy, an optical isomer thereof, or a pharmaceutically acceptable salt thereof .
The method according to claim 1,
Wherein R &lt; ^{1 &gt;} is -H or -OH, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.
A step of culturing Streptomyces sp. KCB13F003 strain to obtain a strain culture (step 1);
Extracting the strain culture obtained in step 1 to obtain an extract (step 2); And
(3) a step of purifying the extract obtained in the step (2).
An antimicrobial composition comprising a compound represented by the general formula (1) of claim 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
6. The method of claim 5,
Wherein the bacterium is Staphylococcus aureus or Salmonella typhimurium.
An antimicrobial pharmaceutical composition comprising a compound represented by the general formula (1) of claim 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
An antimicrobial feed composition comprising a compound represented by the general formula (1) of claim 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
An antimicrobial cosmetic composition comprising a compound represented by the general formula (1) of claim 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
A strain of Streptomyces sp. KCB13F003 producing the compound of claim 1.

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