KR20180068581A - Macrocyclic lactam-based derivative compound, preparing method and use thereof - Google Patents

Abstract

Provided are a compound represented by any one of chemical formulas 1 to 3, an isomer thereof, a derivative, a solvate, or pharmaceutically acceptable salt thereof; and a production method and a use thereof. According to the present invention, the compound can be used to inhibit the growth of bacteria and antibiotic resistant bacteria using the antimicrobial activity and the antimicrobial activity against antibiotic-resistant bacteria; and to prevent and treat infectious diseases infected by or caused by bacteria such as Sotto disease of silkworms.

Description

Macrocyclic lactam-based derivative compounds, preparing method and use thereof,

Macrolactam derivatives, their production methods and uses.

Bombyx , a larva of the silkworm moth belonging to the Lepidoptera, mori ) is one of the representative insects that have been raised by humans for a long time. Silkworm-related industries accounted for 10.1% of total national exports in the 1960s. However, research on the utilization of silk thread produced by silkworms and silkworms is underway. Silkworms are often used in the form of health supplements. In silkworms, the silkworm, which is a flank of silkworms, the silkworms that died of syphilis in silkworms, and the natus moths, are used as medicines. The silkworm's pupa is also used for food. The silk thread produced by silkworms can be used as a raw material for toothpaste and soap, and can be used as a dental shielding film.

However, silkworms are one of the most highly affected insects, and are also very sensitive to pathogens. Viruses, fungi, and bacteria are present in pathogens causing disease in silkworms. A particular type of bacteria causing bacterial disease in silkworms Bacillus Turin group N-Sys (Bacillus thuringiensis), Pseudomonas Aero carbidopa (Aeromonas caviae), Bacillus spring rain septi kusu (Bacilus bombysepticus , Serratia marcescenes , Pseudomonas aeruginosa , Enterococcus, mundtii ). Rather than using special methods to control a wide variety of bacterial diseases, it is all about thoroughly disinfecting the jams and locks and controlling the breeding environment.

Insect defense mechanisms are largely based on physical defenses that physically interfere with external infringement, and additionally by chemical defenses that correspond to external infringement. For example, Apterostigma < RTI ID = 0.0 > dentigerum ants defend themselves by using anti-fungal substances produced by the symbiotic Pseudonocardia strain (Oh et al., Nat. Chem. Biol. , 2009, 5, 391).

Therefore, it is necessary to search for a novel compound that inhibits bacterial growth from silkworm intestinal microorganisms that are judged to be symbiotic with silkworms.

A macrocyclic lactam derivative compound, an isomer thereof, a derivative, a solvate thereof, or a pharmaceutically acceptable salt thereof.

A macrolactam derivative derivative, an isomer thereof, a derivative thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof.

Derivatives thereof, solvates thereof, or pharmaceutically acceptable salts thereof, in the manufacture of a medicament for use as a medicament.

There is provided an antimicrobial composition comprising a macrocyclic lactam derivative compound, an isomer thereof, a derivative thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof.

There is provided a method for sterilizing or bacterium of a bacterium using a macrocyclic lactam derivative compound, an isomer thereof, a derivative, a solvate, or a pharmaceutically acceptable salt thereof.

There is provided a method for preventing or treating an infectious disease caused by or caused by a bacterium using a macrocyclic lactam derivative compound, an isomer thereof, a derivative, a solvate, or a pharmaceutically acceptable salt thereof.

According to one aspect, there is provided a compound represented by any one of formulas (1) to (3), an isomer thereof, a derivative, a solvate thereof, or a pharmaceutically acceptable salt thereof.

According to another aspect, there is provided a strain of Streptomyces sp. SD53 which produces the above compound, an isomer thereof, a derivative, a solvate, or a pharmaceutically acceptable salt thereof.

According to another aspect, there is provided a method of producing said compound, an isomer, derivative, solvate, or pharmaceutically acceptable salt thereof.

According to another aspect, there is provided an antimicrobial composition comprising the above compound, an isomer thereof, a derivative, a solvate, or a pharmaceutically acceptable salt thereof.

According to another aspect, there is provided a method for sterilizing or bacterium of a bacterium using the compound, an isomer thereof, a derivative, a solvate, or a pharmaceutically acceptable salt thereof.

According to another aspect, there is provided a method for preventing or treating an infectious disease caused by or caused by a bacterium using the compound, an isomer thereof, a derivative, a solvate, or a pharmaceutically acceptable salt thereof.

The compound represented by any one of formulas (1) to (3), an isomer thereof, a derivative, a solvate, or a pharmaceutically acceptable salt thereof has antimicrobial activity and antimicrobial activity against antibiotic resistant bacterium, And can be used to prevent or treat an infectious disease caused by or caused by bacteria, for example, a fatal disease of silkworm.

1 is a photograph of a culture medium in which Streptomyces sp. Strain SD53 was cultured.

One aspect may be at least one compound selected from the group consisting of compounds represented by Chemical Formulas 1 to 3, isomers, derivatives, solvates, or pharmaceutically acceptable salts thereof.

[Chemical Formula 1]

.

.

(2)

.

.

(3)

.

.

Wherein R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ and R ₉ independently represent a hydrogen atom, a hydroxyl group, a substituted or unsubstituted C ₁ to C _A substituted or unsubstituted C ₆ to C ₁₀ aryl group, a substituted or unsubstituted C ₆ to C ₁₀ heterocyclic group, or a combination thereof.

In Formula 2, CY1 is a substituted or unsubstituted heterocyclic ring having 5 or 6 carbon atoms; R ₁ ', R' ₂ , and R ' ₃ are each independently a hydrogen atom, a hydroxyl group, a substituted or unsubstituted C ₁ to C ₅ alkyl group, a substituted or unsubstituted C ₆ to C ₁₀ aryl group, An unsubstituted C ₆ to C ₁₀ heterocyclic group, or a combination thereof.

In Formula 3, CY2 is a substituted or unsubstituted cyclopentenone ring;

는 단일결합 또는 이중결합을 나타낼 수 있다.R " ₁ , R " ₂ and R " ₃ are each independently selected from the group consisting of hydrogen, -SR _a , a substituted or unsubstituted C ₁ to C ₅ alkyl group, a substituted or unsubstituted C ₆ to C ₁₀ aryl group, A substituted or unsubstituted C ₆ to C ₁₀ heterocyclic group, or a combination thereof, wherein R _a is hydrogen, a C ₁ to C ₅ alkyl group, a carboxypropyl group substituted with an amide group, an acetamide group substituted with an acetamide group Carboxypropyl group, or a combination thereof,

May represent a single bond or a double bond.

The compounds represented by the above formulas (1) to (3) are macrocyclic compounds. The compounds represented by the above Formulas 1 to 3 are named Bombyxamycin.

The term "substituted" as used herein refers to a substituent that is introduced in place of a hydrogen atom when at least one hydrogen atom of the organic compound is substituted with another atomic group to form a derivative, and "substituent" refers to an atomic group into which the substituent is introduced.

The "substitution" of the alkyl group, aryl group or heterocyclic group (ring) used in the above Chemical Formulas 1 to 3 is a C ₁ to C ₅ alkyl group substituted with a halogen atom or a halogen atom (for example, CCF ₃ , CHCF ₂ , CH ₂ F, CCl _3, etc.), a hydroxy group, a nitro group, a cyano group, an amino group, an amidino group, an acetamido group, hydrazine, hydrazone, a carboxyl group or a salt thereof, a sulfonic acid group or a salt thereof, phosphoric acid or a salt thereof, or a C ₁ to C _A C ₂ to C ₅ alkenyl group, a C ₂ to C ₅ alkynyl group, a C ₃ to C ₁₀ cycloalkyl group, a C ₆ to C ₁₀ aryl group, a C ₆ to C ₁₀ heteroaryl group, A C ₆ to C ₂₀ arylalkyl group, a C ₆ to C ₂₀ heteroarylalkyl group, or a combination thereof.

The term " alkyl group " used in the above Chemical Formulas 1 to 3 means a fully saturated branched or unbranched (or straight chain or linear) hydrocarbon group.

Non-limiting examples of the "C ₁ to C ₅ alkyl group" include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and n-pentyl. One or more hydrogen atoms may be substituted with a substituent as defined in the above " substitution ".

The "C ₆ to C ₁₀ aryl group" used in the above Chemical Formulas 1 to 3 means an aromatic system containing at least one ring, either alone or in combination, and includes, for example, phenyl, naphthyl, tetrahydro Naphthyl, and the like. Also, at least one hydrogen atom of the aryl group may be substituted with a substituent as defined in the above-mentioned " substitution ".

The "C ₆ to C ₁₀ heterocyclic group (ring)" used in the above Chemical Formulas 1 to 3 is a group (ring) containing at least one hetero atom selected from N, O, P or S, Carbon, and includes, for example, tetrahydropyran, tetrahydropyridine and the like. Also, at least one hydrogen atom of the heterocyclic group (ring) may be substituted with a substituent as defined in the above-mentioned " substitution ".

Wherein R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ and R ₉ independently represent a hydrogen atom, a hydroxyl group, a substituted or unsubstituted C ₁ to C An alkyl group having 1 to ₃ carbon atoms, a substituted or unsubstituted C ₆ heterocyclic group, or a combination thereof.

The compound represented by Formula 1 may include at least one compound selected from the following compounds 1A and 1B.

[Compound 1A]

.

.

[Compound 1B]

.

.

The compound represented by Formula 1 may include one or more compounds selected from the following Compounds 1a and 1b.

[Compound 1a]

.

.

[Compound 1b]

.

.

In Formula 2, CY1 may be a substituted or unsubstituted oxygen-containing heterocyclic ring having 5 carbon atoms.

In Formula 2, R ₁ ', R' ₂ , and R ' ₃ may independently be a hydrogen atom, a hydroxy group, a substituted or unsubstituted C ₁ to C ₃ alkyl group, or a combination thereof.

In Formula 3, R " ₁ , R" ₂ , and R " ₃ may independently be a hydrogen atom, -SR _a , a substituted or unsubstituted C ₁ to C ₃ alkyl group, or a combination thereof. R _a may be hydrogen, a C ₁ to C ₅ alkyl group, a carboxypropyl group substituted with an amide group, a carboxypropyl group substituted with an acetamide group, or a combination thereof.

The compounds represented by the formulas (2) and (3) may include at least one compound selected from the compounds (2A) and (3A).

[Compound 2A]

.

.

[Compound 3A]

.

.

The compound represented by Formula 2 may be Compound 2a:

[Compound 2a]

.

.

The term " isomer " refers to a compound that has the same molecular formula but does not have the same linking or spatial arrangement of constituent atoms in the molecule. Isomers include, for example, structural isomers, and stereoisomers. The stereoisomer may be a diasteromer or an enantiomer. An enantiomer is an isomer that does not overlap with its mirror image, such as the relationship between the left and right hands, and is also referred to as an optical isomer. Enantiomers are divided into R (Rectus) and S (sinister) counterclockwise when four or more substituents are different on the chiral center carbon. Diastereomers are stereoisomers that are not mirror images of each other, and the spatial arrangement of atoms is different and can be divided into the cis-trans isomers.

The term " derivative " refers to a compound obtained by substituting a part of the structure of the above compound with another atom or an atomic group.

The term " solvate " refers to a compound that is solvated in an organic or inorganic solvent. The solvate is, for example, a hydrate.

The term " pharmaceutically acceptable salts " refers to the inorganic and organic acid addition salts of the compounds. The pharmaceutically acceptable salt may be a salt that does not cause serious irritation to the organism to which the compound is administered and does not impair the biological activity and properties of the compound. The inorganic acid salt may be a hydrochloride, a bromate, a phosphate, a sulfate, or a disulfide. The organic acid salt may be at least one selected from the group consisting of a formate, a acetate, an acetate, a propionate, a lactate, an oxalate, a tartrate, a malate, a maleate, a citrate, a fumarate, a beateate, a camphateate, an edisleite, trichloroacetic acid, There may be mentioned acetic acid salts, benzoic acid salts, gluconic acid salts, methanesulfonic acid salts, glycolic acid salts, succinic acid salts, 4-toluenesulfonic acid salts, Or aspartate. The metal salt may be a calcium salt, a sodium salt, a magnesium salt, a strontium salt, or a potassium salt.

Another aspect provides Streptomyces sp. SD53 strain.

The strain may produce a compound represented by any one of formulas (1) to (3), a stereoisomer, a derivative thereof, or a pharmaceutically acceptable salt thereof.

The strain includes its variants. The mutant may be, for example, a mutant caused by a natural mutation or an anthropogenic mutation. Anthropogenic mutations can be caused by chemical mutagens such as ultraviolet light, physical mutagens, or base compounds.

The strain includes a spore, a bacterium, or a culture thereof.

The strain may be isolated or derived from intestinal flora of a silkworm.

In another aspect, there is provided a method for isolating a strain of Streptomyces sp. SD53 (Accession No .: KCTC13167BP); And isolating a compound represented by any one of formulas (1) to (3), a stereoisomer, a derivative or a pharmaceutically acceptable salt thereof from a culture of said strain, A method for producing a compound, a stereoisomer, derivative, or pharmaceutically acceptable salt thereof, is provided.

The method includes culturing Streptomyces sp. SD53 strain (Accession No. KCTC13167BP). The step of culturing may be to culture the strain in a liquid medium or a solid medium. The medium may include as a carbon source, for example, glucose, starch syrup, dextrin, starch, molasses, animal oil, vegetable oil, or a combination thereof. The medium may include, for example, wheat bran, soybean meal, wheat, malt, cottonseed, fish, corn steep liquor, gravy, yeast extract, ammonium sulfate, sodium nitrate, urea, or combinations thereof as the source of nitrogen.

The culture may be culturing under aerobic conditions with shaking or standing. The incubation temperature may be, for example, about 20 째 C to about 37 째 C, about 26 째 C to about 35 째 C, or about 30 째 C.

The method comprises separating a compound represented by any one of Chemical Formulas 1 to 3, a stereoisomer, a derivative thereof, or a pharmaceutically acceptable salt thereof from a culture.

The step of separating may comprise the step of concentrating, centrifuging, filtering, or performing chromatography on the culture. The chromatography can be, for example, column chromatography, planar chromatography, paper chromatography or thin film chromatography depending on the form of stationary phase. Chromatography can be, for example, gas chromatography, liquid chromatography, or affinity chromatography, depending on the physical properties of the mobile phase. The liquid chromatography may be, for example, high performance liquid chromatography (HPLC). Chromatography can be, for example, according to a separation method, ion exchange chromatography, size-exclusion chromatography. The chromatography may be, for example, normal chromatography or reverse phase chromatography.

Another aspect provides an antimicrobial composition comprising a compound represented by any one of formulas (1) to (3), a stereoisomer thereof, a derivative thereof, or a pharmaceutically acceptable salt thereof.

The composition may have an antibacterial activity against microorganisms. The composition may have an antimicrobial activity against, for example, a pathogenic microorganism or a non-pathogenic microorganism. The composition may have antimicrobial activity against Gram-positive bacteria or Gram-negative bacteria. Wherein the composition is Staphylococcus (Staphylococcus aureus), Enterococcus faecalis nose kusu par (Enterococcus faecalis), and Enterobacter nose kusu par when Titanium (Enterococcus faecium), Proteus Howe serie (Proteus which may have antimicrobial activity against a bacterium consisting of hauseri , Klebsiella pneumoniae , Salmonella enterica , Escherichia coli , and Bacillus thuringiensis . have.

The composition may have an antimicrobial activity against an antibiotic-resistant bacterium. The term " antibiotic resistance " refers to a drug resistance that a microorganism can survive even when exposed to an antibiotic, and an antibiotic resistant bacterium refers to a bacterium having antibiotic resistance. The antibiotic-resistant bacterium may be a bacterium that is resistant to methicillin, vancomycin, or a combination thereof. Bacteria resistant to methicillin include, for example, methicillin-resistant Saccharomyces aureus (Methicillin-resistant Saccharomyces aureus) MRSA). The bacterium resistant to vancomycin may be, for example, a vancomycin-resistant enterococcus (VRE).

The composition may further comprise known active ingredients having antimicrobial activity against microorganisms.

The composition may be a pharmaceutical composition, a disinfecting composition, a feed composition, or a combination thereof. The pharmaceutical composition includes medicines and quasi-drugs.

The pharmaceutical composition may further comprise a carrier, an excipient or a diluent. Carriers, excipients and diluents include, for example, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Vaginal cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil.

The pharmaceutical compositions may be formulated in the form of powders, granules, tablets, capsules, oral preparations such as suspensions, emulsions, syrups and aerosols, external preparations, suppositories or sterilized injection solutions according to conventional methods. In the case of formulation, it can be prepared using diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, surfactants and the like which are usually used.

In the above pharmaceutical composition, the solid preparation for oral administration may be a tablet, a pill, a powder, a granule, or a capsule. The solid preparation may further comprise an excipient. The excipient may be, for example, starch, calcium carbonate, sucrose, lactose, or gelatin. In addition, the solid preparation may further comprise a lubricant such as magnesium stearate or talc. In the above pharmaceutical composition, the liquid preparation for oral administration may be a suspension, a solution, an emulsion, or a syrup. The liquid formulation may comprise water, or liquid paraffin. The liquid preparations may contain excipients such as wetting agents, sweetening agents, perfumes, or preservatives. In the above pharmaceutical composition, the preparation for parenteral administration may be a sterilized aqueous solution, a non-aqueous solvent, a suspension, an oil, a lyophilized or a left-over preparation. Non-aqueous solvents or suspensions may include vegetable oils or esters. The vegetable oil may be, for example, propylene glycol, polyethylene glycol, or olive oil. The ester may be, for example, ethyl oleate. The suppository base may be witepsol, macrogol, tween 61, cacao paper, laurin, or glycerogelatin.

The preferred dosage of the pharmaceutical composition varies depending on the condition and the weight of the individual, the severity of the disease, the drug form, the administration route and the period, but can be appropriately selected by those skilled in the art. However, the compounds, isomers, derivatives, solvates, or pharmaceutically acceptable salts thereof may be administered in a dosage of about 0.0001 mg / kg to about 100 mg / kg, or about 0.001 mg / kg to about 100 mg / The dose may be administered once to 24 times per day, once to 7 times per day to 1 week, or 1 to 24 times per month to 12 months. The compound, its isomer, derivative, solvate, or pharmaceutically acceptable salt in the pharmaceutical composition may be present in an amount of about 0.0001% to about 10%, or about 0.001% to about 1% .

The method of administration may be oral or parenteral administration. The method of administration may be, for example, oral, transdermal, subcutaneous, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, topical, intranasal, intratracheal, or intradermal. The compositions may be administered systemically or locally and may be administered alone or in combination with other pharmaceutically active compounds.

The disinfecting composition may be a disinfectant cleaner, a shower foam, an oral cleanser, a wet tissue, a hand cleaner, a humidifier filler, a mask, an ointment or a filter filler.

The composition for feed can be added to the feed of mammals or insects other than humans. For example, the feed composition can be added to silkworm feed or mulberry leaves through application, spraying, coating, dipping, or a combination thereof.

Another aspect relates to a method for the preparation of a pharmaceutical composition comprising the step of applying, spraying, coating, dipping or combinations thereof in an in vitro, a compound represented by any one of formulas 1 to 3, a stereoisomer, a derivative or a pharmaceutically acceptable salt thereof. A method of sterilizing or bacterium is provided.

The term " applying " refers to applying the composition to the surface of an object. The term " spray " refers to spraying the composition. The term " coating " refers to coating the surface of an object with the composition. The term " immersion " refers to immersing an object in the composition.

The term " sterilization " refers to the action of killing microorganisms. The term " bacterium " refers to an action that inhibits the growth or proliferation of microorganisms.

The method can be carried out by coating, spraying, coating, dipping or a combination thereof with a compound represented by any one of formulas (1) to (3), a stereoisomer, a derivative or a pharmaceutically acceptable salt thereof.

In another aspect, the present invention provides a method for preventing or causing infectious diseases caused by or caused by a bacterium comprising administering to a subject a compound represented by any one of Chemical Formulas 1 to 3, a stereoisomer, derivative, or pharmaceutically acceptable salt thereof Or < / RTI >

The subject may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat or cat. The subject may be an insect, for example a silkworm.

The term " prophylactic " refers to any act that inhibits disease or delays onset by administration of the composition. The term " treatment " refers to any action that improves or alleviates a symptom of a disease by the administration of the composition.

Examples of infectious diseases caused or caused by bacteria include bacteria such as Staphylococcus aureus , Enterococcus faecalis , and Enterococcus faecium , Proteus < RTI ID = 0.0 > Proteus hauseri), Klebsiella pneumoniae in (Klebsiella pneumoniae), Salmonella Entebbe Rica (Salmonella enterica), E. coli (Escherichia E. coli , and Bacillus thuringiensis , which are caused by or caused by a bacterium. The disease may be, for example, food poisoning, skin inflammation, otitis media, cystitis, sepsis, pneumonia, diarrhea, or a combination thereof. The disease may be, for example, sotto disease of the silkworm.

The term " comprising " is used herein to indicate that it is possible to add and / or intervene other elements, not to exclude other elements unless specifically stated otherwise.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these embodiments are for illustrative purposes only, and the scope of the present invention is not limited to these embodiments.

Example  One. Bamboo samaisin  Isolation and identification of its antibacterial activity

One. Streptomyces  genus( Streptomyces species) SD53  Isolation of strain

In order to screen for a strain producing a substance having antimicrobial activity, the SD53 strain was isolated in the field of the 5th day silkworm purchased from a silkworm farm in Bongeun-eup, Bongeun-eup, Chungcheongbuk-do, Various isolation media were used for the isolation of the strain. The SD53 strain was isolated in the separate medium of A1 + C (4 g of yeast extract per liter of L, 2 g of peptone, 10 g of starch, and 1 g of calcium carbonate).

The whole genome of SD53 strain was isolated and 16S ribosomal DNA was cloned by polymerase chain reaction (PCR) and the nucleic acid sequence was analyzed (Genbank Accession No. KY249557, SEQ ID NO: 1).

As a result of the nucleic acid sequence analysis, the SD53 strain was identified as a strain of Streptomyces sp. This strain was named as Streptomyces sp. SD53 strain and the strain was deposited on November 30, 2016 at the Korea Research Institute of Bioscience & Biotechnology.

2. Streptomyces sp. SD53  Culture of strain

Streptomyces sp. SD53 was inoculated into sterilized YEME solid medium (10 g of malt extract, 4 g of yeast extract, 2 g of glucose, and 18 g of agar per 1 L of water) and incubated at about 30 ° C for 1 Followed by tea cultivation.

The SD53 strain was inoculated from 150 ml to 200 ml of YPM liquid medium (2 g of yeast extract, 2 g of peptone, and 4 g of mannitol per 1 L of water) from the sterilized primary culture medium and cultured at 200 rpm Followed by secondary culturing at about 30 DEG C for 2 days while shaking.

25 ml of the second cultured culture was inoculated into 1 L of YPM liquid medium and cultured for 3 days at about 30 째 C with shaking at 160 rpm.

3. Bambic Samaisin of  Separation and purification

SD53 cultures were obtained, which were cultured in the same manner as in Example 1.2.

A stand ring was attached to the stand to fix the Separatory Funnel (capacity 3 L), and 1 L of SD53 culture and 1.5 L of ethyl acetate (Suppol Gold Co.) were injected. The inlet was closed with a stopper, the mixture was stirred for 1 minute, and then the mixture was allowed to stand for 3 minutes so as to be divided into an ethyl acetate layer and a water layer. The water layer was removed from the culture using a separating funnel, and 100 g of anhydrous sodium sulfate (purified water) was added to the ethyl acetate layer to remove residual water in the culture. The resulting ethyl acetate layer was filtered on filter paper (Advantec), transferred to a round flask and concentrated using a concentrator. The remaining water layer was again extracted with 1.5 L of ethyl acetate. This process was repeated to obtain 3 g of crude extract in a total of 20 L of SD53 culture.

The obtained crude extract was divided into six fractions using reverse phase chromatography (Sepak, C ₁₈ resin 20 g, reversed-phase).

The eluates of the six fractions were used by reducing water from 20% in water to 80% (v / v) water, and the last fraction was fractionated in 50% methanol / dichloromethane to give six fractions Ml). The fractions used were 20% (v / v), 40% (v / v), 60% (v / v), 80% (v / v) (v / v) methanol / water solution was used, and an additional 200 mL of 50% (v / v) methanol / dichloromethane solution was used. Fractions were started using a 20% (v / v) methanol / water solution, followed by increasing the concentration of methanol and proceeding with fractionation. After the fraction using the 100% (v / v) methanol / water solution was completed, the fraction was finished using a 50% (v / v) methanol / dichloromethane solution.

Fractions 3 and 4 were then analyzed by liquid chromatogram-mass spectrometry for fractions of 60% (v / v) and 80% (v / v) water / methanol, respectively. The composition of the fractions was analyzed using LC / MS coupled with Agilent Technologies' 1200 series LC and 6130 series mass spectrometer. The mass spectrum was measured using a Q-High resolution ESI mass spectrometer from Thermo Scientific and expressed in terms of mass / charge (m / z). It was confirmed by liquid chromatography-mass spectrometry and hydrogen nuclear magnetic resonance spectroscopy analysis that a new compound was contained in the culture of the strain.

The fractions 3 and 4, which had been concentrated by vacuum drying, were each dissolved in 2 ml of methanol (purified gold) and the lysate was subjected to reversed-phase half-reaction under an isocratic condition of 58% (v / v) acetonitrile / Separation by preparative HPLC column (YMC-Triart C ₁₈ reversed-phase semi-preparative HPLC column, particle diameter 5 mm, 250 x 10 mm (length x inner diameter), elution rate 2 ml / min, UV detector 280 nm) (Kromasil) Respectively. Thereby, about 15 mg of each of the mincedic smaicin A, the bambic smaicin B, and the bambic smaicin C were obtained at an elution time of about 28 minutes, about 21 minutes, and about 11 minutes, respectively. An additional separation procedure was performed to purely separate the bambic smaicin B and the bambic smaicin C. The mixture containing bambic smaicin B was resuspended in 2 ml of methanol and eluted with a gradient of methanol / water from 65% (v / v) to 90% (v / v) Respectively. Under such separation conditions, it was confirmed that pure bambique semahisin B eluted after 36 minutes. The mixture containing bambic smaicin C was also dissolved in 2 ml of methanol and eluted with a gradient of acetonitrile / water from 35% (v / v) to 60% (v / v) Lt; / RTI > Under such separation conditions, it was confirmed that pure bambique sematicin C eluted after 27 minutes. In addition, piceamycin (Dirk Schulz et al., Journal of Antibiotics, 2009, vol.62, pp. 513-518) was isolated under isocratic conditions of 58% (v / v) acetonitrile / water .

4. Bambic Samay  Physicochemical characterization of A to C

The structure of bambic smaximin A to C was determined by nuclear magnetic resonance spectrum. Nuclear magnetic resonance spectrum ( ¹ H NMR, ¹³ C NMR) was 500 MHz NMR of Bruker and DMSO-d ₆ was used as solvent. The mass spectrum was measured using a JMS-700 FAB mass spectrometer from Jeol Corporation and expressed in terms of mass / charge (m / z).

Tables 1 to 3 show the structural localization of the Bambic smaicins A to C by nuclear magnetic resonance spectroscopy.

[Bombyxamycin A]

(1) Molecular formula: C ₂₇ H ₃₃ NO ₃

(2) Molecular weight: 419

(3) Color: Dark yellow

(4) ¹ H-NMR (DMSO-d ₆ , 500 MHz): See Table 1

(5) ¹³ C-NMR (DMSO-d ₆ , 125 MHz): See Table 1

[Bamboo Samaisin B]

(1) Molecular formula: C ₂₇ H ₃₃ NO ₃

(2) Molecular weight: 419

(3) Color: Dark yellow

(4) ¹ H-NMR (DMSO-d ₆ , 500 MHz): See Table 2

(5) ¹³ C-NMR (DMSO-d ₆ , 125 MHz): See Table 2

[Bamboo Samaisin C]

(1) Molecular formula: C ₂₇ H ₃₁ NO ₅

(2) Molecular weight: 449

(3) Color: Pale yellow

(4) ¹ H-NMR (DMSO-d ₆ , 500 MHz): See Table 3

(5) ¹³ C-NMR (DMSO-d ₆ , 125 MHz): See Table 3

Nuclear Magnetic Resonance Spectrum of Bamberg Samaisin A location Bambic Samay  A δ _C , type δ _H , mult  (Hz) J ) One 166.3, C 2 121.3, CH 5.55, d (11.5) 3 133.7, CH 6.82, dd (11.5, 11.5) 4 124.4, CH 7.45, dd (11.5, 11.5) 5 135.2, CH 6.35, dd (11.5, 11.5) 6 121.7, CH 6.60, dd (15.0, 11.5) 7 140.1, CH 6.41, d (15.0) 8 135.1, C 9 131.4, CH 5.52, m 10 37.2, CH ₂ 2.31, m 11 68.5, CH 4.04, m 11-OH 4.77, d (5.0) 12 44.5, CH ₂ 2.86, dd (14.5, 5.0)
2.46, dd (14.5, 7.0) 13 199.8, C 14 132.0, CH 6.15, d (15.5) 15 136.6, CH 7.26, dd (15.5, 11.0) 16 126.6, CH 6.13, dd (11.0, 11.0) 17 132.4, CH 6.81, dd (11.0, 11.0) 18 122.7, CH 6.16, dd (11.0, 11.0) 19 135.1, CH 5.94, dd (11.0, 11.0) 20 127.1, CH 6.69, dd (15.0, 11.0) 21 132.7, CH 6.27, dd (15.0, 11.5) 22 130.1, CH 6.16, dd (11.5, 11.5) 23 137.7, CH 5.23, dd (11.5, 11.5) 24 30.5, CH 3.12, m 25 44.8, CH ₂ 3.46, m
2.30, m 25-NH 7.96, dd (6.5, 6.5) One' 12.5, CH ₃ 1.64, s 2' 17.9, CH ₃ 0.94, d (6.5)

Nuclear Magnetic Resonance Spectrum Data of Bambic Samaisin B location Bambic Samay  B δ _C , type δ _H , mult  (Hz) J ) One 165.9, C 2 123.3, CH 5.58, d (15.0) 3 130.6, CH 6.59, dd (15.0, 11.5) 4 124.4, CH 6.82, dd (11.5, 11.5) 5 133.3, CH 6.24, dd (11.5, 11.5) 6 120.9, CH 6.49, dd (15.5, 11.5) 7 140.3, CH 6.50, d (15.5) 8 135.7, C 9 129.7, CH 5.84, t (7.5) 10 33.7, CH ₂ 2.19, m 11 43.0, CH ₂ 3.08, m
2.18, m 12 67.3, CH 4.09, m 13 199.1, C 14 130.5, CH 5.99, d (15.5) 15 144.3, CH 7.00, dd (15.5, 11.5) 16 131.2, CH 6.42, dd (11.5, 11.5) 17 136.6, CH 6.84, dd (11.5, 11.5) 18 127.5, CH 6.04, dd (11.5, 11.5) 19 134.6, CH 6.19, dd (11.5, 11.5) 20 127.0, CH 6.39, dd (15.0, 11.5) 21 132.3, CH 6.58, dd (15.0, 11.5) 22 132.3, CH 5.97, dd (11.5, 11.5) 23 137.8, CH 5.19, dd (11.5, 11.0) 24 34.4, CH 2.71, m 25 44.0, CH ₂ 3.36, m
2.74, m 25-NH 7.71, dd (7.0, 7.0) One' 12.3, CH ₃ 1.49, s 2' 17.7, CH ₃ 0.96, d (6.5)

Nuclear Magnetic Resonance Spectrum Data of Bamberg Samaisin C location Bambic Samaisin C δ _C , type δ _H , mult  (Hz) J ) One 165.8, C 2 123.3, CH 5.63, d (11.5) 3 132.3, CH 6.80, dd (11.5, 11.5) 4 126.4, CH 7.41, dd (11.5, 11.5) 5 133.5, CH 6.30, dd (11.5, 11.5) 6 124.6, CH 6.84, dd (15.5, 11.5) 7 138.2, CH 6.16, d (15.5) 8 89.1, C 9 79.4, CH 3.83, dd (5.0, 3.5) 9-OH 5.88, d (3.5) 10 77.5, CH 4.50, dd (5.0, 3.5) 10-OH 6.14, d (3.5) 11 167.6, C 12 100.1, CH 5.08, s 13 189.6, C 14 131.7, CH 6.48, d (15.5) 15 135.8, CH 7.38, dd (15.5, 11.0) 16 127.1, CH 6.09, dd (11.0, 11.0) 17 130.6, CH 6.78, dd (11.5, 11.0) 18 123.3, CH 6.34, dd (11.5, 11.5) 19 134.3, CH 6.10, dd (11.5, 11.5) 20 127.6, CH 6.71, dd (15.0, 11.5) 21 131.8, CH 6.38, dd (15.0, 11.5) 22 129.9, CH 6.14, dd (11.5, 11.5) 23 137.4, CH 5.22, dd (11.5, 11.5) 24 31.3, CH 3.04, m 25 44.8, CH ₂ 3.29, m
2.60, m 25-NH 7.99, dd (6.5, 6.5) One' 18.2, CH ₃ 1.38, s 2' 18.1, CH ₃ 0.95, d (6.5)

The chemical structural formulas of BAMBIC SALAMIN A to C analyzed from nuclear magnetic resonance spectral data are shown below.

[Bamboo Samaishin A]

[Bamboo Samaisin B]

[Bamboo Samaisin C]

5. Bambic Samay  Identification of antimicrobial activity of A to C

The minimum inhibitory concentration (MIC) of the substance was measured in order to measure the bacterial growth inhibition effect of the bambic samaisin A to C.

Specifically, Gram-positive bacteria such as Staphylococcus aureus (ATCC 25923), Enterococcus faecalis , ATCC 19433), and Enterococcus faecium, ATCC 19434), and Gram-negative bacteria of Proteus Howe Seri (Proteus hauseri, ATCC 13315), ah Klebsiella pneumoniae (Klebsiella pneumoniae, ATCC 10031), Salmonella Entebbe Rica (Salmonella enterica, ATCC 14028), and E. coli ( Escherichia coli , ATCC 25922) was prepared.

158 쨉 l of Mueller Hinton broth (MHB) medium was added to the first column of a 96-well microplate, and 80 쨉 l of each medium was added to each column from the 2nd column to the 11th column, and 100 쨉 l was added to the 12th column. At this time, column 11 was used as a growth control and column 12 was used as a sterility control. Ampicillin and tetracycline were dissolved in dimethyl sulfoxide (DMSO) as a positive control, and 12.8 mg / ml of DMSO was used as a negative control. Respectively. 2 μl of the prepared sample was added to the first column, and samples 2 to 10 were subjected to serial dilution.

Thereafter, the concentration of the prepared bacteria was measured using a spectrophotometer, and 20 쨉 l of the bacteria prepared in the concentration of ⁵ × 10 ⁵ CFU / ml were inoculated into the wells 1 to 10. The microplate inoculated with the bacteria was incubated at about 37 캜 for about 16 hours to about 20 hours. The minimum inhibitory concentration (MIC) of the compound was determined by visual observation of the degree of bacterial killing and the results are shown in Table 4.

Antibiotic MIC ([mu] g / ml) Gram-positive bacteria Gram-negative bacteria S. Aureus E. Paekaris E. Paesium P. Haaseri K. pneumoniae S. enterica Escherichia coli Bambic Samaisin A 8 128 > 128 0.5 > 128 One > 128 Bambic Samaisin B 8 > 128 > 128 One > 128 One > 128 Bambic Samaisin C 64 > 128 > 128 > 128 > 128 > 128 > 128 Ampicillin 0.13 0.25 0.25 0.06 > 128 0.25 4 Tetracycline > 128 > 128 > 128 > 128 0.25 > 128 > 128

As shown in Table 4, bambic samaisin A, and bambic samaisin B inhibited the growth of P. hasterii, S. enterica, and S. aureus. In particular, the activity against Gram-negative bacteria such as P. hasterii and S. entericica was superior to the activity against Gram-positive S. aureus. On the other hand, Bamberg semathicin C showed no growth inhibitory effect on Gram-positive bacteria and Gram-negative bacteria. Thus, it was confirmed that Bambic samaisin A and Bamberg samaisin B were excellent in antibacterial activity.

6. Silkworm, a bacterial pathogen Bacillus Turigienesses  About Bambic Samay  Confirming the antimicrobial activity of A to C and fcyamycin

In order to test whether the bambic samaisin A to C and the ficamyicin are effective in controlling the bacterial disease of silkworms, a test for the pathogenic bacterium Bacillus thuringiensis , which causes sotto disease, The growth inhibitory effects of bambic smaximin A to C and fcyanhydride were measured.

Bacillus thuringiensis belongs to gram-positive bacteria and has the characteristic of producing crystal toxin. When silkworms ingest toxins produced by Bacillus thuringiensis, the ingested toxins are dissolved by the alkaline digestive juices of the silkworms, absorbed through the barrier and paralyzed. The silkworms that ingest toxins cause acute symptoms like pesticide poisoning. Within 1 hour of toxin ingestion, the mulberry leaves are stopped and the action stops, releasing the digestive juices through diarrhea and vomiting, and then fainting.

The antimicrobial activity of bacillus thuringiensis , bacillus thuringiensis , and bambic smaesins A to C was measured by the method described in Example 1. 5, and the results are shown in Table 5.

Antibiotic MICs (μg / mL) Bacillus Turinigansis Fiche iaxin 0.19 Bambic Samaisin A 26 Bambic Samaisin B 18 Bambic Samaisin C ND ^*

* ND: Not determined

As shown in Table 5, fishiamycin, bambic smaicin A, and bambic smaicin B inhibited the growth of B. turinigansis while the bambic smaicin C showed no growth inhibitory effect. Accordingly, it has been reported that fishiamycin, bambic smaeicin A, and bambic smaicin B are excellent in antimicrobial activity against B. turiniganis causing silkworm disease of silkworms and can be used for the prophylaxis or treatment of silkworm disease Respectively.

7. Antibiotic resistant bacteria Fishy  Identify antimicrobial activity

It was confirmed whether ficheximine had a growth inhibitory effect on bacteria resistant to the antimicrobial agent.

Saccharomyces aureus resistant to the antibiotic Methicillin, and Enterococcus sp. Resistant to Vancomycin were prepared in the same manner as described in Example 1.5. ) Bacterium. The results are shown in Table 6. < tb > < TABLE >

Gram-positive bacteria MIC (μg / mL) Piceamycin S. aureus M126 (MRSA *) 0.125 S. aureus 98-11-R782 (MRSA *) 0.0625 S. aureus 96-10-2432 0.25 S. aureus 96-10-2626 0.125 S. aureus smith (smith) 0.125 S. aureus ATCC 25923 0.25 E. Paesium 00-5-U1211 (VRE **) One E. Paekaris 99-2-P442 (VRE **) 0.25 E. Paekaris ATCC 29212 One E. Paesium ATCC 10541 One

*: Methicillin-resistant S. aureus: MRSA)

**: Vancomycin-resistant Enterococcus (VRE)

As shown in Table 6, ficheximine had excellent antimicrobial activity against methicillin-resistant S. aureus and vancomycin-resistant enterococcus as well as Gram-positive bacteria. Therefore, it was confirmed that ficarcissin is excellent in antimicrobial activity against bacteria, especially antibiotic-resistant bacteria, and that ficamycin can be used for the prevention or treatment of antibiotic-resistant bacteria.

Korea Biotechnology Research Institute KCTC13167BP 20161130

<110> SEOUL NATIONAL UNIVERSITY R & DB Foundation <120> Macrocyclic lactam-based derivative compound, preparing method          and use thereof <130> GN-116060-KR <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 987 <212> DNA <213> Artificial Sequence <220> Nucleic acid encoding 16S rRNA of Streptomyces sp. SD53 strain <400> 1 tagttggtgg ggtaatggcc taccaaggcg aacgacgggt agccggcctg agagggcgac 60 cggccacact gggactgaga cacggcccag actcctacgg gaggcagcag tggggaatat 120 tgcacaatgg gcgaaagcct gatgcagcga cgccgcgtga gggatgacgg ccttcgggtt 180 gtaaacctct ttcagcaggg aagaagcgaa agtgacggta cctgcagaag aagcgccggc 240 taactacgtg ccagcagccg cggtaatacg tagggcgcaa gcgttgtccg gaattattgg 300 gcgtaaagag ctcgtaggcg gcttgttgcg tcggttgtga aagcccgggg cttaaccccg 360 ggtctgcagt cgatacgggc aggctagagt gtggtagggg agatcggaat tcctggtgta 420 gcggtgaaat gcgcagatat caggaggaac accggtggcg aaggcggatc tctgggccat 480 tactgacgct gaggagcgaa agcgtgggga gcgaacagga ttagataccc tggtagtcca 540 cgccgtaaac gttgggaact aggtgttggc gacattccac gtcgtcggtg ccgcagctaa 600 cgcattaagt tccccgcctg ggggagtacg gccgcaaggc taaaactcaa aggaattgac 660 gggggcccgc acaagcagcg gagcatgtgg cttaattcga cgcaacgcga agaaccttac 720 caaggcttga catataccgg aaagcatcag agatggtgcc ccccttgtgg tcggtataca 780 ggtggtgcat ggctgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag 840 cgcaaccctt tgttctgtgt tgccagcatg cccttccggg gtgatgggga ctcaaaggaa 900 actgccgggg tcaacttcgg aggaaggtgg gggacgacgt caagtcatca tgccccttat 960 gtcttgggct ggcacacgtg ctacaat 987

Claims (20)

At least one compound selected from the group consisting of compounds represented by Chemical Formulas 1 to 3, isomers, derivatives, solvates, or pharmaceutically acceptable salts thereof:
[Chemical Formula 1]

In Formula 1,
R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ and R ₉ independently represent a hydrogen atom, a hydroxyl group, a substituted or unsubstituted C ₁ to C ₅ alkyl group, Or an unsubstituted C ₆ to C ₁₀ aryl group, a substituted or unsubstituted C ₆ to C ₁₀ heterocyclic group, or a combination thereof.
(2)

In Formula 2,
CY1 is a substituted or unsubstituted heterocyclic ring having 5 or 6 carbon atoms;
R ₁ ', R' ₂ , and R ' ₃ are each independently a hydrogen atom, a hydroxy group, a substituted or unsubstituted C ₁ to C ₅ alkyl group, a substituted or unsubstituted C ₆ to C ₁₀ aryl group, An unsubstituted C ₆ to C ₁₀ heterocyclic group, or a combination thereof.
(3)

In Formula 3,
CY2 is a substituted or unsubstituted cyclopentenone ring;
R " ₁ , R " ₂ and R " ₃ are each independently selected from the group consisting of hydrogen, -SR _a , a substituted or unsubstituted C ₁ to C ₅ alkyl group, a substituted or unsubstituted C ₆ to C ₁₀ aryl group, A substituted or unsubstituted C ₆ to C ₁₀ heterocyclic group, or a combination thereof, wherein R _a is hydrogen, a C ₁ to C ₅ alkyl group, a carboxypropyl group substituted with an amide group, an acetamide group substituted with an acetamide group Carboxypropyl group, or a combination thereof,

Represents a single bond or a double bond. A compound according to claim 1, wherein R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ and R ₉ independently represent a hydrogen atom, A substituted or unsubstituted C ₁ to C ₃ alkyl group, a substituted or unsubstituted C ₆ heterocyclic group, or combinations thereof. The compound according to claim 1, wherein in the general formula (2), CY1 is a substituted or unsubstituted oxygen-containing heterocyclic ring having 5 carbon atoms. 2. The compound according to claim 1, wherein R ₁ ', R' ₂ , and R ' ₃ are independently a hydrogen atom, a hydroxy group, a substituted or unsubstituted C ₁ to C ₃ alkyl group, or combinations thereof. 3. The compound according to claim 1, wherein R " ₁ , R " ₂ and R " ₃ are independently a hydrogen atom, -SR _a , a substituted or unsubstituted C ₁ to C ₃ alkyl group, , Wherein R _a is hydrogen, a C ₁ to C ₅ alkyl group, a carboxypropyl group substituted with an amide group, or a carboxypropyl group substituted with an acetamide group. The compound according to claim 1, wherein the compound represented by Formula (1) comprises at least one compound selected from the following Compounds 1A and 1B:
[Compound 1A]

,
[Compound 1B]

. The compound according to claim 1, wherein the compound represented by Formula 1 comprises at least one compound selected from the following Compounds 1a and 1b:
[Compound 1a]

,
[Compound 1b]

. The compound according to claim 1, wherein the compound represented by Formula 2 and Formula 3 is at least one compound selected from Compound 2A and Compound 3A:
[Compound 2A]

,
[Compound 3A]

. The compound according to claim 1, wherein the compound represented by Formula 2 is Compound 2a:
[Compound 2a]

. Streptomyces sp. SD53 strain (accession number: KCTC13167BP) which produces the compound of claim 1, an isomer thereof, a derivative, a solvate, or a pharmaceutically acceptable salt thereof. Culturing strain Streptomyces sp. SD53 (Accession No: KCTC13167BP); And
Isolating the compound of claim 1, an isomer, derivative, solvate, or pharmaceutically acceptable salt thereof from a culture of said strain, the isomer, derivative, solvate or pharmaceutically acceptable salt of said compound To yield an acceptable salt. An antimicrobial composition comprising the compound of claim 1, an isomer, derivative, solvate, or pharmaceutically acceptable salt thereof. 13. The composition of claim 12, wherein the composition is a pharmaceutical composition, a disinfecting composition, a feed composition, or a combination thereof. The composition according to claim 12, wherein the composition has antimicrobial activity against gram-positive bacteria or gram-negative bacteria. The composition according to claim 12, wherein the composition is selected from the group consisting of Staphylococcus aureus , Enterococcus faecalis), and Enterobacter nose kusu Titanium (Enterococcus faecium) when par, Proteus Howe serie (Proteus hauseri), Klebsiella pneumoniae (Klebsiella in wherein the composition has antimicrobial activity against bacteria consisting of pneumoniae , Salmonella enterica , Escherichia coli , and Bacillus thuringiensis . The composition of claim 12, wherein the composition has an antimicrobial activity against an antibiotic-resistant bacterium. 16. The composition of claim 16, wherein the antibiotic-resistant bacterium is a bacterium that is resistant to methicillin, vancomycin, or a combination thereof. A method of sterilizing a bacterium, comprising applying, spraying, coating, immersing, or a combination thereof in vitro, a compound of claim 1, an isomer, derivative, solvate, or pharmaceutically acceptable salt thereof Or bacterium. 19. The method of claim 18, wherein the method comprises applying, spraying, coating, dipping, or a combination thereof to the silkworm. A method for preventing or treating an infectious disease caused or caused by a bacterium comprising administering to a subject a compound of claim 1, an isomer, derivative, solvate, or pharmaceutically acceptable salt thereof.

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