Novel cinnamic acid-containing cyclic peptide compound and use thereof

Abstract

The present invention relates to a novel cinnamic acid-containing cyclic peptide compound and a use thereof. Gwanacinnamycin, which is a novel substance produced by the Kitasatospora sp. GA02 strain according to the present invention, was found to have excellent Mycobacterium tuberculosis inhibitory activity and be stable in vivo, and thus to have potential for use as an antituberculous agent.

Description

Novel cinnamic acid-containing cyclic peptide compound and uses thereof

[0001] The present invention relates to a novel cylindrical peptide compound containing cinnamic acid and its uses.

[0002]

[0003] Tuberculosis is a fatal infectious disease caused by Mycobacterium tuberculosis, a bacterium belonging to the genus Mycobacteria, and is an acute and chronic infectious disease that has threatened human health for thousands of years. Approximately 85% of all cases of tuberculosis occur in the lungs, and the disease can metastasize to other organs via the bloodstream or lymphatic vessels. The primary route of transmission is through the airborne dispersal of respiratory secretions from patients, and major symptoms include a cough accompanied by bloody sputum, chills, night sweats, and weight loss.

[0004] The treatment of tuberculosis is characterized by the need to administer various antibiotics over a long period. As of 2007, there were approximately 13.7 million chronic patients, mainly in developing countries, with an estimated 9.3 million new cases and 1.8 million deaths annually. Currently, rifampicin and isoniazid, which act specifically on the cell walls of mycobacteria, are used as first-line treatments, and resistance to these drugs is referred to as multidrug-resistant tuberculosis.

[0005] In South Korea, tuberculosis is designated as a legal infectious disease, with approximately 170,000 patients reported as of 2010 and about 35,000 new cases occurring annually. In particular, due to the increasing trend of multidrug-resistant tuberculosis patients, there is an urgent need for the development of anti-tuberculosis drugs with new mechanisms of action.

[0006]

[0007] Accordingly, the inventors of the present invention developed a novel cyclic peptide compound containing cinnamic acid and confirmed its potential for use as an antituberculosis agent, thereby completing the present invention.

[0008]

[0009] One aspect provides a compound represented by the following chemical formula 1, a stereoisomer thereof, a solvate, or a pharmaceutically acceptable salt:

[0010] [Chemical Formula 1]

[0011] .

[0012] Another aspect provides a strain of the genus Kitasaphosphora (Kitasatosporasp.) GA02 (accession number: KCTC16224BP) that produces the above compound or a stereoisomer thereof.

[0013] Another aspect provides a method for producing said compound or its stereoisomer, comprising the steps of: culturing a strain of the genus *Gitasaphosphora* GA02 (accession number: KCTC16224BP); and isolating said compound or its stereoisomer from a culture, concentrate, extract, or dried product of said strain.

[0014] Another aspect provides a pharmaceutical composition for the prevention or treatment of bacterial infections of the genus Mycobacterium, comprising the above compound, stereoisomers thereof, solvates, or pharmaceutically acceptable salts.

[0015] Another aspect provides a health functional food composition for preventing or improving bacterial infections of the genus Mycobacterium, comprising the above compound, its stereoisomer, solvate, or salt.

[0016] Another aspect provides a method for preventing or treating bacterial infections of the genus Mycobacterium, comprising the step of administering the compound, its stereoisomer, solvate, or pharmaceutically acceptable salt to an individual.

[0017] Other objects and advantages of this application will become more apparent from the following detailed description, together with the appended claims and drawings. Anything not described in this specification is omitted, as it can be sufficiently recognized and inferred by those skilled in the art of this application or a similar art field.

[0018]

[0019] The present invention relates to a novel cylindrical peptide compound containing cinnamic acid and its use. The cylindrical peptide compound containing cinnamic acid produced by the genus *G.A. 2* strain of the present invention has inhibitory activity against Mycobacterium tuberculosis. Therefore, it can be used as an antituberculosis agent.

[0020]

[0021] Figure 1 shows the genus G.A. 22 strain of *G.A. 2222* isolated from sediments of the mountainous soil of Gwanaksan.

[0022] Figure 2 shows the inactivation curves when tuberculosis bacteria are treated with Gwanak cinnamycin at different concentrations.

[0023] Figures 3a, 3b, and 3c show the results of the cytotoxicity analysis of Gwanak cinnamycin.

[0024]

[0025] Each description and embodiment disclosed in this application may be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in this application fall within the scope of this application. Furthermore, the scope of this application should not be considered limited by the specific descriptions provided below.

[0026]

[0027] One aspect provides a compound represented by the following chemical formula 1, a stereoisomer thereof, a solvate, or a pharmaceutically acceptable salt:

[0028] [Chemical Formula 1]

[0029] .

[0030] The compound represented by the above chemical formula 1, Gwanacinnamycin, may be derived from the genus Kitasaphosporasp. GA02 strain.

[0031] The compound represented by Chemical Formula 1 above is a cyclic peptide containing cinnamic acid, and a protecting group or a modifying group may be attached to the side chain amino acids of the peptide to obtain chemical stability, enhanced pharmacological properties (half-life, absorption, potency, efficacy, etc.), altered specificity (e.g., broad spectrum of biological activity), and reduced antigenicity. The cyclic peptide may include an acetyl group, a fluorenylmethoxycarbonyl group, a palmitoyl group, polyethylene glycol (PEG), etc. Additionally, the peptide may optionally include a targeting sequence, a tag, a labeled residue, or an amino acid sequence prepared for a specific purpose to increase half-life or peptide stability.

[0032] The term "isomer" in "stereoisomer" refers to a compound that has the same molecular formula but differs in the way constituent atoms are connected or in their spatial arrangement within the molecule. Isomers may include, for example, structural isomers and stereoisomers. The stereoisomers mentioned above may be diastereomers or enantiomers. Enantiomers refer to isomers that do not overlap with their mirror images, much like the relationship between a left hand and a right hand, and are also called optical isomers. Enantiomers are classified as R (Rectus: clockwise) and S (Sinister: counterclockwise) when there are four or more different substituents on the chiral central carbon. Diastereomers are stereoisomers that are not mirror images of each other, and can be divided into cis and trans isomers due to differences in the spatial arrangement of atoms.

[0033] The term "solvate" refers to a compound solvated in an organic or inorganic solvent. The solvate may be, for example, a hydrate, but is not specifically limited thereto.

[0034] The term "salt" refers to an inorganic or organic acid addition salt of a compound. The pharmaceutically acceptable salt may be a salt that does not cause severe irritation to the organism to which the compound is administered and does not impair the biological activity and physical properties of the compound. The inorganic salt may be a hydrochloride, bromate, phosphate, sulfate, or disulfate, but is not particularly limited thereto. The organic salt may be a formate, acetate, propionate, lactate, oxalate, tartrate, malate, maleate, citrate, fumarate, besylate, camsylate, edicilate, trichloroacetic acid, trifluoroacetate, benzoate, gluconate, methanesulfonate, glycolate, succinate, 4-toluenesulfonate, galacturonate, emvonate, glutamate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, or aspartate. The metal salts mentioned above may be calcium salts, sodium salts, magnesium salts, strontium salts, or potassium salts, but are not particularly limited thereto.

[0035]

[0036] Another aspect provides a strain of the genus Kitasaphosporasp. GA02 that produces a compound represented by Chemical Formula 1 or a stereoisomer thereof. The same parts described above apply equally to the strain.

[0037] The above strain of the genus Gitasatospora belongs to the Actinobacteria.

[0038] The strain may have a 16S rRNA gene having at least 95%, 96%, 97%, 98%, 99%, 99.5%, or 99.9% sequence identity or sequence homology with SEQ ID NO. 1. Specifically, the strain may have the 16S rRNA gene of SEQ ID NO. 1.

[0039] The above strain includes its variants. The variants may be, for example, variants produced by natural mutations or artificial mutations. Artificial mutations may be caused by physical mutagens, such as ultraviolet rays, or chemical mutagens, such as base compounds.

[0040] The above strain includes the spores, cells, or cultures of the strain, concentrates, extracts, or dried products thereof.

[0041] The above strain may be isolated or derived from soil sediments in a mountainous area, specifically, it may be isolated or derived from soil sediments in the Gwanaksan mountainous area.

[0042] The strain mentioned above may be isolated. In this specification, the term "isolated" means that which is not found in nature but is artificially isolated and available for use.

[0043] The strain above may have antituberculosis activity, specifically, it may produce Gwanacinnamycin, which has antituberculosis activity. In addition, the strain above may exhibit antibacterial activity against Mycobacterium tuberculosis.

[0044] The term "anti-tuberculosis" refers to any action that suppresses or inhibits tuberculosis bacteria, or treats and prevents tuberculosis. The aforementioned tuberculosis bacteria may refer to tuberculosis bacteria belonging to the genus Mycobacterium, specifically Mycobacterium tuberculosis, and more specifically Mycobacterium tuberculosis (mc). 2 It may mean 6230 strains, but is not limited thereto.

[0045] The term "antimicrobial" refers to any action that inhibits the growth or survival of microorganisms. The aforementioned microorganisms may refer to bacteria belonging to the genus Mycobacterium, specifically Mycobacterium tuberculosis, and more specifically Mycobacterium tuberculosis (mc). 2 It may mean 6230 strains, but is not limited thereto.

[0046] In one embodiment, it was confirmed that Gwanak cinnamycin, a novel substance produced by the genus Kitasapospora GA02 strain isolated from Gwanaksan Mountain, has excellent anti-tuberculosis effects and in vivo stability, thus confirming its potential for use as an anti-tuberculosis agent.

[0047]

[0048] Another aspect provides a method for producing said compound or its stereoisomer, comprising the steps of: culturing a strain of the genus Kitasaphospora (Kitasatosporasp.) GA02 (accession number: KCTC16224BP); and isolating said compound or its stereoisomer from a culture of said strain, or from a concentrate, extract, or dried product thereof. The same parts as described above apply equally to said method.

[0049] The above method includes the step of culturing the genus Kitasatospora GA02 strain (accession number: KCTC16224BP).

[0050] The above-mentioned culturing step may involve culturing the strain in a liquid medium or a solid medium. The medium may include, for example, glucose, starch syrup, dextrin, starch, molasses, animal oil, or vegetable oil as a carbon source. The medium may include, for example, wheat bran, soybean meal, wheat, malt, cottonseed meal, fish meal, corn starch, meat juice, yeast extract, ammonium sulfate, sodium nitrate, or urea as a nitrogen source.

[0051] Culture may be performed under aerobic conditions with shaking or standing. The culture temperature may be, for example, 20°C to 40°C, 25°C to 37°C, 28°C to 35°C, or about 30°C. The culture time may be, for example, about 1 day to about 2 months, about 1 day to about 6 weeks, about 1 day to about 1 month, about 1 day to about 2 weeks, or about 1 day to about 1 week.

[0052] The above method includes the step of isolating a compound represented by Formula 1, or a stereoisomer thereof, from a culture of a strain, a concentrate, an extract, or a dried product thereof.

[0053] The term "concentrate" refers to a concentrated form of the culture, and the term "extract" refers to an extract obtained from the culture or its concentrate; it may include a diluted or concentrated extract, a dried product obtained by drying the extract, a modified or purified product thereof, or a fraction obtained by fractionating the extract. Additionally, in this specification, the term "dried product" may refer to a dried form of the culture, its concentrate, its extract, or its fraction; the drying method may be air drying, natural drying, spray drying, or freeze-drying, but is not particularly limited thereto.

[0054] The above separation step may include steps of concentrating, centrifuging, filtering, or performing chromatography on the culture medium. Chromatography may be, for example, column chromatography, planar chromatography, paper chromatography, or thin-layer chromatography depending on the form of the stationary phase. Chromatography may be, for example, gas chromatography, liquid chromatography, or affinity chromatography depending on the physical properties of the mobile phase. Liquid chromatography may be, for example, high-performance liquid chromatography (HPLC). Chromatography may be, for example, ion exchange chromatography or size-exclusion chromatography depending on the separation method. Chromatography may be, for example, normal-phase chromatography or reverse-phase chromatography.

[0055]

[0056] Another aspect provides a pharmaceutical composition for the prevention or treatment of bacterial infections of the genus Mycobacterium, comprising a compound represented by Chemical Formula 1, a stereoisomer thereof, a solvate, or a pharmaceutically acceptable salt thereof. The same parts as described above apply equally to the composition.

[0057] The term "bacterial infection of the genus Mycobacterium" refers to all infectious diseases caused by bacteria of the genus Mycobacterium. The said bacterial infection of the genus Mycobacterium may be selected from the group consisting of tuberculosis, leprosy, and nontuberculous mycobacteria (NTM) infections, and specifically may be tuberculosis.

[0058] The bacteria of the genus Mycobacterium mentioned above refer to bacteria belonging to the family Mycobacteriaceae of the class Actinobacteria. Bacteria of the genus Mycobacterium may be acid-fast. The bacteria of the genus Mycobacterium mentioned above may be Mycobacterium tuberculosis, nontuberculous mycobacteria, or BCG (Bacillus Calmette-Guerin) strains. The above-mentioned bacteria of the genus Mycobacterium may be, for example, Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium abscessus, Mycobacterium africanum, Mycobacterium microti, Mycobacterium leprae, Mycobacterium canetti, Mycobacterium avium, Mycobacterium kansasii, or Mycobacterium marinum.

[0059] The above composition may exhibit antibacterial activity against bacteria of the genus Mycobacterium, and specifically may exhibit antibacterial activity against Mycobacterium tuberculosis.

[0060] The term "tuberculosis" refers to a contagious bacterial infection caused by Mycobacterium tuberculosis. It is a serious infectious disease that primarily infects the lungs and causes chronic respiratory illness, and it can be transmitted through the respiratory system.

[0061] The above-mentioned tuberculosis may be multidrug-resistant (MDR) or extensive drug-resistant (XDR) tuberculosis. The above-mentioned multidrug-resistant tuberculosis may be tuberculosis in which the tuberculosis bacteria are not killed even when isoniazid and rifampicin are administered to the human body. Extensive drug-resistant tuberculosis may be tuberculosis that is resistant not only to isoniazid and rifampicin but also to quinolones and injectable drugs simultaneously.

[0062] The term "prevention" refers to any act of suppressing a disease or delaying its onset through the administration of a composition.

[0063] The term "treatment" refers to any act of improving or beneficially altering the symptoms of a disease through the administration of a composition.

[0064] The above pharmaceutical composition may further include an antibiotic. The antibiotic may be an antituberculosis drug. The above anti-tuberculosis agents are, for example, ethambutol, isoniazid, rifampicin, SQ-109, pyrazinamide, streptomycin, kanamycin, capreomycin, ethionamide, prothionamide, enviomycin, para-aminosalicylic acid, cycloserine, amikacin, levofloxacin, moxifloxacin, gatifloxacin, ofloxacin, terizidone, thionamide, and ethionamide. It may be protionamide, clofazimine, linezolid, amoxicillin, clavulanate, thioacetazone, imipenem, cilastatin, clarithromycin, bedaquiline, delamanid, limipenem, cilastatin, meropenem, or a combination thereof. The compound represented by Formula 1, its stereoisomers, solvates, or pharmaceutically acceptable salts and the antibiotic may be a single or individual composition for simultaneous or sequential administration.

[0065] The above pharmaceutical composition may further comprise a carrier, an excipient, or a diluent. The carrier, excipient, and diluent may include, for example, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil.

[0066] The above pharmaceutical compositions may each be formulated according to conventional methods into oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, as well as topical preparations, suppositories, or sterile injectable solutions. When formulating, they may be prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

[0067] In the above pharmaceutical composition, the solid dosage form for oral administration may be a tablet, pill, powder, granule, or capsule. The solid dosage form may further include an excipient. The excipient may be, for example, starch, calcium carbonate, sucrose, lactose, or gelatin. Additionally, the solid dosage form may further include a lubricant such as magnesium stearate or talc. In the above pharmaceutical composition, the liquid dosage form for oral administration may be a suspension, liquid formulation, emulsion, or syrup. The liquid dosage form may include water or liquid paraffin. The liquid dosage form may include an excipient, for example, a humectant, a sweetener, a flavoring agent, or a preservative. In the above pharmaceutical composition, the formulation for parenteral administration may be a sterile aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a freeze-dried or suppository. The non-aqueous solvent or suspension may include a vegetable oil or an ester. The vegetable oil may be, for example, propylene glycol, polyethylene glycol, or olive oil. The ester may be, for example, ethyl oleate. The base of the suppository may be witepsol, macrogol, tween 61, cacao oil, laurin oil, or glycerogelatin.

[0068] The preferred dosage of the above pharmaceutical composition may vary depending on the individual's condition and body weight, the severity of the disease, the form of the drug, the route of administration, and the duration, but can be appropriately selected by a person skilled in the art. However, the compound, its isomers, derivatives, solvates, or pharmaceutically acceptable salts may be administered, for example, in amounts of about 0.0001 mg / kg to about 100 mg / kg, or about 0.001 mg / kg to about 100 mg / kg, divided into 1 to 24 doses per day, 1 to 7 doses per week from 2 days to 1 year, or 1 to 24 doses per month to 12 months. In the above pharmaceutical composition, the compound, its isomers, derivatives, solvates, or pharmaceutically acceptable salts may be included in an amount of about 0.0001 weight% to about 10 weight%, or about 0.001 weight% to about 1 weight%, based on the total weight of the composition.

[0069] The method of administration may be oral or parenteral. The method of administration may be, for example, oral, transdermal, subcutaneous, rectal, intravenous, intra-arterial, intraperitoneal, intramuscular, intrasternal, local, intranasal, intratracheal, or intradermal. The composition may be administered systemically or locally, and may be administered alone or in combination with other pharmaceutically active compounds.

[0070]

[0071] Another aspect provides a health functional food composition for the prevention or improvement of tuberculosis comprising a compound represented by Chemical Formula 1, a stereoisomer thereof, a solvate, or a salt thereof. The same as described above applies equally to the composition.

[0072] The term "improvement" includes all actions that cause the symptoms of a disease to improve or become beneficial.

[0073] The above health functional food may be used as a functional food or added to various foods by encapsulating, powdering, or suspension of a compound represented by Chemical Formula 1, its stereoisomer, solvate, or salt. The above foods are, for example, meat, sausage, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, functional foods, and health foods.

[0074]

[0075] Another aspect provides a method for preventing or treating tuberculosis, comprising the step of administering to an individual a compound represented by Chemical Formula 1, a stereoisomer thereof, a solvate, or a pharmaceutically acceptable salt thereof. The same parts as described above apply equally to the method.

[0076] The above-mentioned individual may be a mammal, for example, a human, cattle, horses, pigs, dogs, sheep, goats, or cats, and specifically refers to mammals excluding humans. The above-mentioned individual may be an individual diagnosed with tuberculosis or one with a high probability of being diagnosed with tuberculosis.

[0077] The method of administration may be oral or parenteral. The method of administration may be, for example, oral, transdermal, subcutaneous, rectal, intravenous, intra-arterial, intraperitoneal, intramuscular, intrasternal, local, intranasal, intratracheal, or intradermal. The pharmaceutical composition may be administered systemically or locally, and may be administered alone or in combination with other pharmaceutically active compounds.

[0078] The preferred dosage of the above pharmaceutical composition may vary depending on the patient's condition and body weight, the severity of the disease, the form of the drug, the route of administration, and the duration, but can be appropriately selected by a person skilled in the art. The dosage may be, for example, within the range of about 0.001 mg / kg to about 100 mg / kg, about 0.01 mg / kg to about 10 mg / kg, or about 0.1 mg / kg to about 1 mg / kg based on an adult. The administration may be once daily, multiple times daily, once a week, once every two weeks, once every three weeks, or once every four weeks to once a year.

[0079]

[0080] The present invention will be explained in more detail below through examples. However, these examples are intended to illustrate the invention and the scope of the invention is not limited to these examples.

[0081]

[0082] Example 1. Isolation of the genus Kitasaphosporasp. GA02 strain

[0083] In 2019, strain GA02 of the genus Kitasaphospora was isolated from sediments of the mountain soil of Gwanaksan Mountain, located in Gwanak-gu, Seoul, South Korea.

[0084] Specifically, to isolate the strain, ISP2 medium (yeast extract 4 g, malt extract 10 g, glucose 4 g, agar powder 18 g / 1 L sterilized water) was used. Subsequently, based on the results of 16S rRNA sequencing analysis, the strain was identified as a strain of the genus Kitasaphosphora (Kitasatosporasp.). The strain was named Kitasaphosphora strain GA02 (Genbank Accession No. PQ740319) and deposited at the Korea Culture Collection Center (KCTC) of the Korea Research Institute of Biotechnology and Bioengineering under accession number KCTC16224BP. The 16S rRNA gene sequence of the GA02 strain is shown as Sequence No. 1 in Table 1 below.

[0085]

[0086]

[0087]

[0088] Example 2. Culture and extraction of strain GA02 of the genus *Gitasaphospora*

[0089] The GA02 strain of the genus *Gitasophora* from Experimental Example 1 was inoculated from a solid medium into 50 mL of ISP2 liquid medium and cultured with stirring at 180 rpm and 30°C for 3 days. Subsequently, 10 mL of the culture solution was inoculated into 250 mL of sterilized ISP2 liquid medium. After culturing for 3 days under the same conditions, 20 mL of the culture solution was inoculated into 1 L GSS liquid medium (10 g of soluble starch, 20 g of glucose, 25 g of soy peptone, 1 g of beef extract, 4 g of yeast extract, 2 g of sodium chloride, 2 g of calcium carbonate / 1 L sterilized water) and cultured at 170 rpm and 30°C for 5 days.

[0090] To obtain the substance produced by the genus *G. GA02* strain of *G. GA02*, organic extraction was performed on the culture medium after mass culture was completed using ethyl acetate (EtOAc) in an amount of 1.5 to 2 times the volume of the culture medium. After thoroughly shaking the culture medium and ethyl acetate using a separatory funnel, the mixture was allowed to stand to allow the water layer and the organic layer to separate sufficiently, and then the lower water layer was removed. Residual moisture was removed from the organic layer using anhydrous sodium sulfate, and then the ethyl acetate layer containing the substance produced by the genus *G. GA02* strain was dried under reduced pressure. As a result, 5 g of dried extract was obtained by culturing a total of 20 L.

[0091]

[0092] Example 3. Isolation and Purification of Gwanak Cinnamycin

[0093] According to Experimental Example 2, methanol (MeOH) and Celite were added to the dried extract, and the extract was adsorbed onto Celite by vacuum drying, and fractionation was carried out. In an open column packed with ODS (C18) resin, solvents prepared with methanol / water (w / w) compositions of 20%, 40%, 60%, 80%, and 100% were used sequentially to remove impurities in the first step. A novel substance produced by the genus *G.A. 2* strain of *G.A. 2* was detected in the 100% methanol fraction.

[0094] Subsequently, vacuum drying was performed for further purification, and semi-preparative HPLC (high-performance liquid chromatography) was used to obtain a pure novel substance. Specifically, for sample injection, the 100% methanol fraction was dissolved in methanol, and then separated using reverse-phase chromatography gradient conditions (at least 30 minutes in 30-60% CH3CN-H2O containing 0.1% formic acid; flow rate: 2 mL / min, UV detection: 210 nm) with a Kromasil column (250 x 10 mm, C18, 5 μm), and it was confirmed that the novel substance eluted at 17 minutes. Accordingly, the separated and purified novel substance was named Gwanacinnamycin.

[0095]

[0096] Example 4. Physicochemical properties of Gwanak cinnamycin

[0097] The structure and stereostructure of Gwanak cinnamycin were confirmed based on NMR spectra, and its physicochemical properties are as follows.

[0098]

[0099] [Chemical Formula 1] Structure of Gwanacinnamycin

[0100]

[0101] (1) Molecular formula: C 64 H 88 N 14 O 13

[0102] (2) Molecular weight: 1261

[0103] (3) Color: Yellow

[0104] (4) 1H-NMR (DMSO-d6, 800 MHz): See Table 1

[0105] (5) 13C-NMR (DMSO-d6, 200 MHz): See Table 1

[0106]

[0107]

[0108]

[0109] Experimental Example 1. Evaluation of the anti-tuberculosis activity of Gwanak Cinnamycin

[0110] The antituberculosis effect of Gwanacinnamycin was evaluated.

[0111] Specifically, Mycobacterium tuberculosismc based on H37Rv 2 Strain 6230 was inoculated into Middlebrook 7H9 broth (Difco) supplemented with 0.2% glycerol, 0.05% Tween 80, and 10% albumin dextrose catalase (ADC), and treated with Gwanak cinnamycin at various concentrations. The antibacterial effect was confirmed by resazurin microplate assay. For this purpose, the final Mycobacterium tuberculosis concentration was measured in a 96-well plate using OD. 600 The concentration was adjusted to 0.005, and Gwanak cinnamycin, moxifloxacin (MFX), rifampicin (RIF), and ethambutol (EMB) were treated using a twofold dilution method. Subsequently, a 96-well plate was cultured in a 37°C incubator for approximately 7 days. On the 7th day of culture, 10 μl of resazurin solution (0.0025% wt / vol) was added to each well, and the plates were incubated overnight in a 37°C incubator.

[0112] The following day, the fluorescence intensity of the conversion from resazurin to risofurin metabolites was measured using a SpectraMax® M3 Multi-Mode Microplate Reader (Molecular Devices, Sunnyvale, CA, USA) at excitation / emission (560 / 590 nm). To determine the minimum inhibitory concentration (MIC), a dose response curve was constructed using GraphPad Prism software (version 6.05; San Diego, CA, USA).

[0113] As a result, as shown in Figure 2, at a concentration of Gwanak cinnamycin of 1.1 μM, Mycobacterium tuberculosismc 2 It was confirmed that 50% of the 6230 strains were killed (MIC50 = 1.1 μM). Since the MIC50 value of Gwanak cinnamycin is lower than that of the existing antituberculosis drug ethambutol, it can be seen that Gwanak cinnamycin has excellent inhibitory activity against tuberculosis bacteria.

[0114]

[0115] Experimental Example 2. Evaluation of Cytotoxicity of Gwanak Cinnamycin

[0116] Since Mycobacterium tuberculosis is an intracellular bacterium that inhabits host cells, the cytotoxicity of Gwanak cinnamycin was evaluated. For this purpose, bedaquiline (BDQ) was used as the reference compound, and Trinton X-100 1% was used as the toxic substance.

[0117] Specifically, cytotoxicity of Mouse Bone Marrow Derived Macrophages (mBMDM) extracted from mouse leg bones and differentiated after treatment with Gwanak cinnamycin at various concentrations via a twofold dilution method was measured using the CellTiter 96® AQueous One Solution Cell Proliferation Assay kit. Syto60-stained mBMDMs were observed using a fluorescence microscope (SpectraMax® M3 Multi-Mode Microplate Reader, Molecular Devices, Sunnyvale, CA, USA) and cell counts were measured. Images were captured using the ImageXpress Pico Automated Cell Imaging System (Molecular Devices, Sunnyvale, CA, USA), and the number of viable macrophages was quantified using CellReporterXpress® Image Acquisition and Analysis Software (Molecular Devices, Sunnyvale, CA, USA). Statistical significance was determined through ANOVA analysis comparing the treatment group and the DMSO control group (****P< 0.0001; ns, not significant).

[0118] As a result, as shown in Figure 3, it was confirmed that Gwanak cinnamycin had no cytotoxicity up to the maximum measured concentration of 50 μM in mBMDM, the host cell of Mycobacterium tuberculosis. That is, 1.1 μM is the concentration capable of killing 50% of Mycobacterium tuberculosis, and since Gwanak cinnamycin was harmless up to 50 μM according to the cytotoxicity evaluation results, it was confirmed that Gwanak cinnamycin has in vivo safety.

[0119]

[0120] In summary, Gwanacinnamycin, a novel substance produced by the GA02 strain of the genus Kitasaphospora isolated from Gwanaksan Mountain, has been confirmed to have excellent anti-tuberculosis effects and in vivo stability, thus confirming its potential for use as an anti-tuberculosis agent.

[0121]

[0122] The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will understand that other specific forms can be easily modified without altering the technical spirit or essential features of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

[0123]

[0124]

Claims

1. A compound represented by the following chemical formula 1, its stereoisomers, solvates, or pharmaceutically acceptable salts: [Chemical Formula 1] .

2. A strain of the genus Kitasaphosphora (Kitasatosporasp.) GA02 (accession number: KCTC16224BP) that produces the compound of Claim 1 or a stereoisomer thereof.

3. The strain of claim 2, wherein the strain exhibits antibacterial activity against Mycobacterium tuberculosis.

4. A step of culturing the Kitasaphosphora genus GA02 strain (accession number: KCTC16224BP); and A method for producing a compound represented by Formula 1 or a stereoisomer thereof, comprising the step of isolating a compound represented by Formula 1 or a stereoisomer thereof from a culture of the strain, a concentrate thereof, an extract thereof, or a dried product thereof.

5. A pharmaceutical composition for the prevention or treatment of bacterial infections of the genus Mycobacterium, comprising a compound represented by the following chemical formula 1, a stereoisomer thereof, a solvate, or a pharmaceutically acceptable salt: [Chemical Formula 1] .

6. A pharmaceutical composition according to claim 5, wherein the bacterial infection of the genus Mycobacterium is tuberculosis.

7. A pharmaceutical composition according to claim 5, wherein the composition exhibits antibacterial activity against Mycobacterium tuberculosis.

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