KR101721665B1 - Amide-based derivative compound, preparing method and use thereof - Google Patents

Abstract

An amide-based derivative compound, a production method thereof, and uses thereof. Since the compound has anticancer activity, the compound can be used for preventing or treating cancer.

Description

Amide-based derivative compounds, production methods and uses thereof,

Amide-based derivative compounds, their production methods and uses.

Physiologically active substances derived from microorganisms were generally the source of antibacterial, antifungal or anticancer drugs, and they have been developed as new drugs for the treatment of various diseases including these and have become templates for the development of new drugs. Antibiotics derived from microorganisms are, for example, amphotericin, erythromycin, streptomycin, tetracycline, and vancomycin. In 2003, daptomycin isolated from actinomycetes Streptomyces was approved by the US Food and Drug Administration (FDA) as a next-generation antibiotic. On the other hand, anticancer drugs derived from bacteria include, for example, doxorubicin, bleomycin, mithramycin, neocarzinostatin, pentostatin, epothilone, have.

In order to develop new anticancer drugs structurally, it is necessary to select microorganisms producing useful biologically active substances, and to search for and discover new compounds produced by them.

Therefore, it is necessary to screen for new bacteria and search for new compounds having anticancer activity therefrom.

An amide-based derivative compound, an isomer thereof, a derivative thereof, or a pharmaceutically acceptable salt thereof.

An isomer, a derivative or a pharmaceutically acceptable salt thereof.

Amide derivative compounds, isomers, derivatives or pharmaceutically acceptable salts thereof.

The present invention provides a pharmaceutical composition for preventing or treating cancer comprising an amide derivative compound, an isomer thereof, a derivative thereof, or a pharmaceutically acceptable salt thereof.

The present invention provides a method of preventing or treating cancer using an amide derivative compound, an isomer thereof, a derivative thereof, or a pharmaceutically acceptable salt thereof.

According to one aspect, there is provided a compound represented by formula 1, an isomer thereof, a derivative thereof, or a pharmaceutically acceptable salt thereof.

According to another aspect, there is provided a Streptomyces strain HK10 strain which produces a compound represented by the above formula 1, an isomer thereof, a derivative thereof, or a pharmaceutically acceptable salt thereof.

According to another aspect, there is provided a method for producing a compound represented by the above-mentioned formula 1, an isomer thereof, a derivative thereof, or a pharmaceutically acceptable salt thereof.

According to another aspect, there is provided a pharmaceutical composition for preventing or treating cancer, which comprises a compound represented by Formula 1, an isomer thereof, a derivative thereof, or a pharmaceutically acceptable salt thereof.

According to another aspect, there is provided a method of preventing or treating cancer using the compound represented by the above-mentioned formula 1, an isomer thereof, a derivative thereof, or a pharmaceutically acceptable salt thereof.

Amide derivatives, isomers, derivatives or pharmaceutically acceptable salts thereof have antitumor activity and can be used for the prevention or treatment of various types of cancer.

1 is a photograph of a culture medium in which Streptomyces sp. Strain HK10 was cultured.

An aspect provides a compound represented by formula 1, an isomer, derivative or pharmaceutically acceptable salt thereof:

[Formula 1]

.

.

The compound represented by the formula 1 is an amide-based derivative compound. The compound represented by the above formula 1 is named Salternamide. The amide-based derivative compound may be a compound having two carbon chains bonded to a hexagonal ring at a meta position. Each ring may include, for example, a carbonyl group. Each ring may contain one to three double bonds. For example, a six-membered ring may contain one or two double bonds. The carbon chain may be bonded at positions 2 and 4 of the hexagonal ring, respectively. 6 The carbon chain bonded to the 2-position of each ring can be bonded to the hexagonal ring by an amide bond. The carbon chain may be a substituted or unsubstituted, saturated or unsaturated carbon chain. For example, the carbon chain bonded at the 2-position of the hexagonal ring may be a substituted or unsubstituted unsaturated carbon chain. For example, the carbon chain bonded to the 4-position of the hexagonal ring may be a substituted or unsubstituted saturated carbon chain.

Wherein R ₁ , R ₂ and R ₃ independently represent hydrogen, a hydroxyl group, a halogen group, a cyano group, -C (═O) R _a , -C (═O) OR _a , -OCO _{oR a), -C = N (} R a), -SR a, -S (= O) R a, -S (= O) ₂ R _a alkyl groups, substituted or unsubstituted C ₁ -C _20, substituted Or an unsubstituted C ₁ -C ₂₀ alkoxy group, a substituted or unsubstituted C ₂ -C ₂₀ alkenyl group, a substituted or unsubstituted C ₂ -C ₂₀ alkynyl group, a C ₂ -C ₂₀ alkylene oxo aryloxy oxide group, a substituted or unsubstituted C ₃ -C ₃₀ cycloalkyl group, a substituted or unsubstituted C ₆ -C ₃₀ aryl group, a substituted or unsubstituted C ₆ -C _30, a substituted or unsubstituted C ₆ -C ₃₀ heteroaryl groups, or combinations thereof. Wherein R _a can be hydrogen, a C ₁ -C ₁₀ alkyl group, or a C ₆ -C ₂₀ aryl group. The dashed line in the formula 1 may represent an optional double bond. The dotted line may be a single bond or a double bond. However, in the formula 1, R ₁ and R ₂ may be excluded from forming a ring by fusing with each other.

In the above formula 1, R ₁ , R ₂ , and R ₃ may be, independently of each other, hydrogen, a hydroxyl group, a halogen group, a substituted or unsubstituted C ₁ -C ₂₀ alkyl group, or a combination thereof. The halogen group may be -Cl, -F, -Br, or -I. The C ₁ -C ₂₀ alkyl group may be, for example, a C ₁ -C ₁₅ alkyl group, a C ₁ -C ₁₀ alkyl group, a C ₁ -C ₅ alkyl group, a C ₁ -C ₄ alkyl group, a methyl, , Butyl, or isobutyl.

The compound represented by the formula 1 may be a compound represented by the formula 2:

[Formula 2]

.

.

In the formula 2, R ₁ is hydrogen, a hydroxy group, a halogen group, a cyano _{group, -C (= O) R a} , -C (= O) OR a, -OCO (OR a), -C = N (R a ), -SR _a , -S (= O) R _a , -S (= O) ₂ R _a , A substituted or unsubstituted C ₁ -C ₂₀ alkyl group, a substituted or unsubstituted C ₁ -C ₂₀ alkoxy group, a substituted or unsubstituted C ₂ -C ₂₀ alkenyl group, a substituted or unsubstituted C ₂ -C _A C ₂ -C ₂₀ alkylene oxide group, a substituted or unsubstituted C ₃ -C ₃₀ cycloalkyl group, a substituted or unsubstituted C ₆ -C ₃₀ aryl group, a substituted or unsubstituted A C ₆ -C ₃₀ aryloxy group, a substituted or unsubstituted C ₆ -C ₃₀ heteroaryl group, or a combination thereof. Wherein R _a can be hydrogen, a C ₁ -C ₁₀ alkyl group, or a C ₆ -C ₂₀ aryl group.

Equation 2 may be Equation 3:

[Formula 3]

.

.

In the formula 2 or 3, R ₁ may be hydrogen or a halogen group. The halogen group may be -Cl, -F, -Br, or -I. When R < _{1 >} is Cl, the compound of formula 2 or formula 3 may be a salt amide A. [ When R < ₁ > is H, the compound of formula 2 or formula 3 may be a salt amide B.

The compound represented by the formula 1 may be a compound represented by the formula 4:

[Formula 4]

.

.

In the formula 4, R ₃ is hydrogen, a hydroxy group, a halogen group, a cyano _{group, -C (= O) R a} , -C (= O) OR a, -OCO (OR a), -C = N (R a ), -SR _a , -S (= O) R _a , -S (= O) ₂ R _a , A substituted or unsubstituted C ₁ -C ₂₀ alkyl group, a substituted or unsubstituted C ₁ -C ₂₀ alkoxy group, a substituted or unsubstituted C ₂ -C ₂₀ alkenyl group, a substituted or unsubstituted C ₂ -C _A C ₂ -C ₂₀ alkylene oxide group, a substituted or unsubstituted C ₃ -C ₃₀ cycloalkyl group, a substituted or unsubstituted C ₆ -C ₃₀ aryl group, a substituted or unsubstituted A C ₆ -C ₃₀ aryloxy group, a substituted or unsubstituted C ₆ -C ₃₀ heteroaryl group, or a combination thereof. Wherein R _a can be hydrogen, a C ₁ -C ₁₀ alkyl group, or a C ₆ -C ₂₀ aryl group. In the above formula (4), the hydroxy group at the 4-position may be an (S) or an (R) configuration. In the formula (4), the hydroxy group at the 5-position may be an (S) or an (R) configuration.

Equation 4 may be Equation 5:

[Formula 5]

.

.

In the above formula 4 or formula 5, it may be hydrogen or a substituted or unsubstituted C ₁ -C ₂₀ alkyl group. The C ₁ -C ₂₀ alkyl group may be, for example, a C ₁ -C ₁₅ alkyl group, a C ₁ -C ₁₀ alkyl group, a C ₁ -C ₅ alkyl group, a C ₁ -C ₄ alkyl group, a methyl, , Butyl, isobutyl. When R < ₃ > is H, the compound of formula 4 or formula 5 may be a salt amide C. When R < ₃ > is methyl, the compound of formula 4 or formula 5 may be a salt amide D. [

The alkenyl groups may be C ₂ to C ₂₀ , C ₅ to C ₂₀ , C ₁₀ to C ₂₀ , or C ₁₀ to C ₁₅ alkenyl groups. The alkaryl carbonyl group may be an alkaryl carbonyl date C ₂ to C _20, C ₅ to C _20, C ₁₀ to C _20, or C ₁₀ to C _15.

"Substitution" in the above "substituted" means that a hydrogen atom in the organic compound is substituted for another atomic group to form a derivative to be introduced instead of a hydrogen atom, and "substituent" means an introduced atomic group. Substituents can include, for example, a hydroxy group, a halogen atom, C ₁ to C An alkyl group of _C20, aryl group of C ₂ to C ₂₀ alkenyl groups, C ₂ to C ₂₀ alkynyl group, C ₁ to C ₂₀ heterocycloalkyl group, C ₆ to C ₂₀ aryl group, C ₆ to C ₂₀ of the, C ₆ to C ₂₀ heteroaryl group, or C ₆ to C ₂₀ heteroarylalkyl group, a nitro group, a cyano group, an amino group, an amidino group, hydrazine, hydrazone, a carboxyl group or a salt thereof, a sulfonic acid group or a salt thereof, phosphoric acid or his of It can be a salt.

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 bonsai. Isomers include, for example, structural isomers, and stereoisomers.

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 "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 salt may be, for example, an inorganic acid salt, an organic acid salt, or a metal salt. The inorganic acid salt may be a hydrochloride, a bromate, a phosphate, a sulfate, or a disulfide. Organic acid salts include those derived from organic acids such as formate, acetate, acetate, propionate, lactate, oxalate, tartrate, malate, maleate, citrate, fumarate, bevylate, camylate, eddylate, trichloroacetic acid, Examples of the salt include salts of benzoic acid, salts of gluconic acid, salts of methanesulfonic acid, salts of glycolic acid, succinic acid salts, 4-toluenesulfonic acid salts, glucuronic acid salts, embronic acid salts, glutamic acid salts, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, , 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 a Streptomyces strain HK10 strain (accession number: KCTC12832BP) which produces a compound of the above formula 1, an isomer thereof, a derivative thereof, or a pharmaceutically acceptable salt thereof.

The compounds, isomers, derivatives, and pharmaceutically acceptable salts of the above formula 1 are as described above.

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 salt water or salt earth.

In another aspect, there is provided a method for culturing Streptomyces sp. HK10 strain (Accession No. KCTC12832BP); And isolating a compound of formula 1, an isomer, a derivative thereof, or a pharmaceutically acceptable salt thereof from said culture, to produce a compound of formula 1, an isomer, derivative or pharmaceutically acceptable salt thereof ≪ / RTI >

The compounds, isomers, derivatives, and pharmaceutically acceptable salts of the above formula 1 are as described above.

The method comprises culturing Streptomyces sp. HK10 strain (Accession No. KCTC12832BP).

The Streptomyces sp. HK10 strain is as described above.

The step of culturing may be to culture the strain in a liquid medium or a solid medium. The medium may comprise artificial sea salt. The medium may include, for example, glucose, rice, starch, dextrin, starch, molasses, animal oil, or vegetable oil as a carbon source. The nitrogen source may include, for example, yeast extract, peptone, wheat bran, soybean meal, wheat, malt, cottonseed, fish, cones triticale, juice, ammonium sulfate, sodium nitrate, or urea. The culture medium may contain, if necessary, inorganic salts which promote salt formation, potassium, magnesium, cobalt, chlorine, phosphoric acid, sulfuric acid, or other ions. The medium may include, for example, artificial sea salt, yeast extract, malt extract, or glucose.

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 40 째 C, about 25 째 C to about 37 째 C, about 28 째 C to about 35 째 C, or about 30 째 C. The incubation time may be, for example, from 1 day to 2 months, from 1 day to 6 weeks, from 1 day to 1 month, from 1 day to 2 weeks, or from 1 day to 1 week.

The method includes isolating the compound of formula 1, an isomer, derivative, or pharmaceutically acceptable salt thereof from a culture. Among the cultures, the compound of Formula 1, an isomer thereof, a derivative thereof, or a pharmaceutically acceptable salt thereof can be isolated from Streptomyces sp. HK10 strain. The compound of Formula 1, an isomer thereof, a derivative thereof, or a pharmaceutically acceptable salt thereof may be isolated from a culture solution containing no Streptomyces sp. HK10 strain among the cultures.

Separation of the compound of Formula 1, its isomer, derivative, or pharmaceutically acceptable salt from the culture may include concentration, centrifugation, filtration, or chromatography of the culture. For example, the culture can be extracted with ethyl acetate, water, or a combination thereof. The resulting concentrate can be fractionated by chromatography to yield six fractions depending on the polarity. The chromatography may be reversed phase chromatography using, for example, water, methanol, dichloromethane, or a combination thereof as a mobile phase. The fractions eluted with water / methanol having a volume ratio of 20% of the obtained fractions can be subjected to high performance liquid chromatography (HPLC) to separate the peptide compounds. The HPLC can be carried out using a reverse phase semi-preparative HPLC column with 45% water / acetonitrile as the mobile phase.

Another aspect provides a pharmaceutical composition for preventing or treating cancer comprising a compound of formula 1, an isomer thereof, a derivative thereof, or a pharmaceutically acceptable salt thereof.

The compounds, isomers, derivatives, and pharmaceutically acceptable salts of the above formula 1 are as described above.

The cancer may be, for example, cancer of the liver, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, bacterial sarcoma, acute myelogenous leukemia, chronic myelogenous leukemia, acute lymphocytic leukemia, Cancer of the gallbladder, cancer of the gallbladder, bile duct cancer, colon cancer, retinoblastoma, choroidal melanoma, bladder cancer of the bladder, bladder cancer, peritoneal cancer, pituitary cancer, adenocarcinoma, pituitary adenocarcinoma, papillary adenocarcinoma, Neoplasm, neuroblastoma, neuroblastoma, neuroblastoma, renal cancer, malignant soft tissue tumor, malignant bone tumor, malignant bone tumor, malignant neoplasm, Lymphoma, malignant mesothelioma, malignant melanoma, ovarian tumor, vulvar cancer, urethral cancer, unknown primary site, gastric lymphoma, gastric cancer, gastric carcinoma, gastrointestinal stromal tumor, Wilm's tumor, Cancer of the uterus, cancer of the uterine endometrium, uterine sarcoma, prostate cancer, metastatic brain tumor, rectal cancer, rectal carcinoid tumor, vaginal cancer, spinal cord tumor, neuroendocrine tumor, pancreatic cancer, salivary gland cancer, tonsillar cancer, breast cancer, sarcoma, penile cancer, , Squamous cell carcinoma, lung cancer, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, laryngeal cancer or a combination thereof. The cancer may be, for example, lung cancer, colon cancer, gastric cancer, leukemia, liver cancer, or breast cancer.

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.

The pharmaceutical composition may further comprise known active ingredients having anticancer activity.

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

The pharmaceutical composition may have an oral or parenteral dosage form. The pharmaceutical compositions can be formulated in the form of powders, granules, tablets, capsules, oral preparations such as suspensions, emulsions, syrups and aerosols, external preparations, suppositories or sterile 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 pharmaceutical composition may contain an effective amount of the compound of formula 1, its isomer, derivative, or pharmaceutically acceptable salt. The effective amount may be appropriately selected depending on the cell or individual selected by a person skilled in the art. The severity of the cancer, the age, weight, health, sex, sensitivity of the patient to the drug, time of administration, route of administration and rate of excretion, duration of treatment, elements including drugs used in combination with or co- May be determined by factors well known in the medical arts. Wherein said effective amount is from about 10 mg to about 10 g, from about 50 mg to about 9 g, from about 100 mg to about 8 g, from about 200 mg to about 7 g, from about 300 mg to about 6 g, from about 400 from about 500 mg to about 4 g, from about 500 mg to about 3 g, from about 500 mg to about 2 g, or from about 500 mg to about 1 g. The dosage of the pharmaceutical composition may be, for example, from about 0.001 mg / kg to about 100 mg / kg, from about 0.01 mg / kg to about 10 mg / kg, or from about 0.1 mg / kg to about 1 mg / kg, and may be administered once a day, once a day, or once a day.

Another aspect provides a method of preventing or treating cancer comprising administering to a subject a pharmaceutical composition for the prophylaxis or treatment of cancer comprising a compound of formula 1, an isomer thereof, a derivative, or a pharmaceutically acceptable salt thereof .

The compound, isomer, derivative, pharmaceutically acceptable salt, cancer, prevention, treatment, and pharmaceutical composition of the above formula 1 are as described above.

The subject may be a mammal, including a mouse, a mouse, a dog, a cow, a horse, a monkey, and a human.

The preferred dosage of the compound of formula 1, its isomer, derivative, or pharmaceutically acceptable salt will vary depending on the condition and body weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, . However, the peptide compound can be administered, for example, in an amount 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 / It may be administered once to several times in divided doses.

The compounds of formula 1, its isomers, derivatives or pharmaceutically acceptable salts may be administered alone or in combination with other pharmaceutically active compounds.

The pharmaceutical composition may be administered to a mammal, including a mouse, a mouse, a dog, a cow, a horse, a monkey, and a human, in various routes. The method of administration may be oral or parenteral administration. The method of administration can be, for example, rectal or intravenous, muscular, subcutaneous, intrauterine, or intracerebroventricular injection.

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. Soltanamid  Isolation and identification of its anticancer activity

One. Streptomyces  genus( Streptomyces species ) HK10  Isolation of strain

In order to screen for strains producing substances with anticancer activity, the HK10 strain was isolated from the tiller surface soil collected in November 2011 in a salt trough of 445-9 Shinan Toppan Salt Sewage Combination, Sin-yi-myeon, Shinan-gun, Jeollanam-do. Whole genome of HK10 strain was isolated and 16S ribosomal DNA was cloned by polymerase chain reaction (PCR) and the nucleic acid sequence was analyzed (SEQ ID NO: 1, GenBank Accession No. LC013479.1).

As a result of the nucleic acid sequence analysis, the HK10 strain was identified as a novel strain that was phylogenetically similar to Streptomyces radiopugnans . This strain was named as HK10 strain of Streptomyces, and the strain was deposited on June 4, 2015 in the Korean Collection for Type Cultures (accession number: KCTC12832BP).

2. Streptomyces sp. HK10  Culture of strain

Streptomyces sp. Strain HK10 was inoculated into sterilized YEME solid medium (10 g of malt extract, 4 g of yeast extract, 2 g of glucose, 33 g of artificial sea salt, and 18 g of agar per 1 L of water) Lt; RTI ID = 0.0 > 27 C < / RTI > for several days.

HK10 strain was cultured from the primary cultured solid medium in a sterilized 50 ml of A1 + YPM liquid medium (10 g of starch per liter, 6 g of yeast, 4 g of peptone, 4 g of mannitol, and 33 g of artificial sea Salt) and secondary cultured at about 30 째 C for 2 days while shaking at 200 rpm.

10 ml of the second cultured culture was inoculated into 1 L of A1 + YPM liquid medium and secondary cultured at about 30 DEG C for 5 days while shaking at 180 rpm.

3. Soltanamid  Separation and purification

A culture HK10 cultured in Example 1.2 was obtained.

A stand ring was attached to the stand, and a separatory funnel (capacity 3 L) was fixed, and 1 L of HK10 culture and 1 L of ethyl acetate (CPC Co., Ltd.) were injected. The inlet was closed with a stopper, the mixture was stirred for 30 seconds, and then 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 and 100 g of Anhydrous Sodium Sulfate (Precipitation Gold Co.) 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 L of ethyl acetate. This procedure was repeated to obtain 5 g of crude extract in a total of 60 L of HK10 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).

Fraction 4 of 20% (v / v) water / methanol fraction was analyzed by liquid chromatogram-mass spectrometry and hydrogen nuclear magnetic resonance spectroscopy. 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 the culture of the strain contained new secondary metabolites.

The fraction 4 concentrated by vacuum drying was dissolved in 2 ml of methanol (purified gold) and the lysate was subjected to reverse phase semi-preparative HPLC (HPLC) under isocratic conditions of 45% (v / v) water / acetonitrile (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) (Kromasil). This resulted in 5.0 mg of Solutinamide A, 8.0 mg of Solutinamide B, 20.0 mg of Solutinamide C, and 3.0 mg of Solutin A, respectively, at an elution time of about 28 minutes, about 25 minutes, about 21 minutes, and about 29 minutes, Amide D. < / RTI >

4. Saltenamide  Physicochemical characterization of A to D

Solanthamides A to D were pale yellow oily, stable at room temperature and well soluble in methanol, a medium organic solvent. Solutinamide A tended to be denatured in an acidic environment.

The structures of the soltanamides A to D were determined by nuclear magnetic resonance spectrum, infrared and ultraviolet spectroscopic data, and high resolution mass spectrometry data. Nuclear magnetic resonance spectrum ( ¹ H NMR, ¹³ C NMR) was performed by 600 MHz or 500 MHz NMR of Bruker, and methanol-d ₄ was used as the solvent. The mass spectra were recorded using a Q-High resolution ESI mass spectrometer from Thermo Scientific and expressed in mass / charge (m / z) form. The infrared spectrum was measured using a Thermo NICOLET iS10 spectrometer. The ultraviolet spectrum was measured using a Lambda 35 UV / VIS spectrometer from Perkin Elmer.

Tables 1 and 2 show the structural positioning of the solutin amides A to D by nuclear magnetic resonance spectroscopy.

[Salternamide A]

(1) Molecular formula: C ₂₃ H ₃₂ ClNO ₅

(2) Molecular weight: 437

(3) Color: Pale yellow

(4) Infrared absorption band (ZnSe): 3329, 2957, 2925, 1711, 1662, 1522 cm ^-1

(5) ¹ H-NMR (MeOD, 500 MHz): See Table 1

(6) ¹³ C-NMR (MeOD, 125 MHz): See Table 1

[Solutin amide B]

(1) Molecular formula: C ₂₃ H ₃₃ NO ₅

(2) Molecular weight: 403

(3) Color: Pale yellow

(4) Infrared absorption band (ZnSe): 3328, 2957, 2925, 1711, 1662, 1522 cm ^-1

(5) ¹ H-NMR (MeOD, 600 MHz): See Table 1

(6) ¹³ C-NMR (MeOD, 150 MHz): See Table 1

[Solutin amide C]

(1) Molecular formula: C ₂₃ H ₃₅ NO ₆

(2) Molecular weight: 421

(3) Color: Pale yellow

(4) Infrared absorption band (ZnSe): 3371, 2959, 2926, 1671, 1520 cm ^-1

(5) ¹ H-NMR (MeOD, 600 MHz): See Table 2

(6) ¹³ C-NMR (MeOD, 150 MHz): See Table 2

[Solutinamide D]

(1) Molecular formula: C ₂₄ H ₃₇ NO ₆

(2) Molecular weight: 435

(3) Color: Pale yellow

(4) Infrared absorption band (ZnSe): 3372, 2958, 2928, 1673, 1518 cm ^-1

(5) ¹ H-NMR (MeOD, 600 MHz): See Table 2

(6) ¹³ C-NMR (MeOD, 150 MHz): See Table 2

The chemical shift values of the nuclear magnetic resonance spectra of the soltan amides A and B location Saltenamide A ^a Saltenamide B ^b δ _c , type δ _H , mult  ( Hz  medium J ) d _C , type δ _H , mult  ( Hz  medium J ) One 176.0, C 181.8, C 2 131.8, C 132.4, C 3 134.7, CH 7.68 d (2.0) 134.1, CH 7.63 d (3.0) 4 72.1, C 71.2, C 5 151.2, CH 7.14 d (2.0) 155.4, CH 6.94 dd (10.0, 3.0) 6a
6b 131.3, C 126.6, CH 6.21 d (10.0) 7 41.7, CH ₂ 1.82 m 41.6, CH ₂ 1.78 m 8 24.5, CH ₂ 1.33 m 24.5, CH ₂ 1.31 m 9 26.4, CH ₂ 1.60 m 26.3, CH ₂ 1.59 m 10 35.1, CH ₂ 2.25 t (7.0) 34.9, CH ₂ 2.25 t (7.0) 11 177.7, C 177.5, C One' 167.5, C 167.4, C 2' 122.6, CH 6.16 d (15.5) 121.3, CH 6.14 d (15.5) 3 ' 153.9, CH 6.79 dd (15.5, 8.0) 153.6, CH 6.78 dd (15.5, 8.0) 4' 35.8, CH 2.33 m 35.7, CH 2.33 m 5'a
5'a 45.5, CH ₂ 1.34 m
1.28 m 45.4, CH ₂ 1.33 m 6 ' 31.5, CH 2.43 m 31.5, CH 2.43 m 7 ' 132.2, CH 4.87 d (9.0) 132.1, CH 4.87 d (9.0) 8' 130.7, C 131.2, C 9 ' 25.9, CH ₃ 1.66 d (1.0) 25.9, CH ₃ 1.66 d (1.0) 10 ' 18.1, CH ₃ 1.61 d (1.0) 18.0, CH ₃ 1.61 d (1.0) 11 ' 21.7, CH ₃ 0.91 d (7.0) 21.6, CH ₃ 0.91 d (7.0) 12 ' 19.8, CH ₃ 1.04 d (7.0) 18.4, CH ₃ 1.03 d (7.0) 2'-CH ₃

a : ¹ H 500 MHz, ¹³ C 125 MHz. b : ¹ H 600 MHz, ¹³ C 150 MHz.

The chemical shift values of the nuclear magnetic resonance spectra of the solan amides C and D location Saltenamide C ^b Saltenamide D ^b δ _c , type δ _H , mult  ( Hz  medium J ) δ _c , type δ _H , mult  ( Hz  medium J ) One 194.1, C 194.4, C 2 133.7, C 133.4, C 3 133.0, CH 7.57 s 132.0, CH 7.53 h 4 73.4, C 73.5, C 5 70.9, CH 3.96 dd (8.0, 4.0) 70.9, CH 3.99 dd (8.0, 4.0) 6a
6b 42.4, CH ₂ 2.81 dd (17.0, 8.0)
2.72 dd (17.0, 4.0) 42.1, CH ₂ 2.84 dd (17.0, 8.0)
2.76 dd (17.0, 4.0) 7 39.3, CH ₂ 1.86 m
1.71 m 39.3, CH ₂ 1.88 m
1.72 m 8 24.5, CH ₂ 1.50 m (13.0, 6.0) 24.6, CH ₂ 1.52 m
1.50 m 9 26.6, CH ₂ 1.67 m
1.62 m 26.9, CH ₂ 1.66 m 10 34.9, CH ₂ 2.32 t (7.0) 35.5, CH ₂ 2.31 t (7.5) 11 177.6, C 178.3, C One' 167.2, C 169.8, C 2' 122.7, CH 6.11 dd (16.0, 1.0) 130.4, CH 3 ' 153.5, CH 6.76 dd (16.0, 7.0) 145.4, CH 6.19 dd (10.0, 1.5) 4' 34.9, CH 2.32 m 32.6, CH 2.55 m 5'a
5'a 45.4, CH ₂ 1.31 ddd (13.5, 9.0, 6.5)
1.28 ddd (13.5, 8.0, 6.0) 46.0, CH ₂ 1.34 m
1.26 m 6 ' 31.4, CH 2.42 dq (9.0, 7.0, 6.0) 31.9, CH 2.39 m 7 ' 132.1, CH 4.85 d (9.0) 132.3, CH 4.88 d (9.0) 8' 131.2, C 131.3, C 9 ' 25.9, CH ₃ 1.65 d (1.0) 26.0, CH ₃ 1.66 d (1.0) 10 ' 18.0, CH ₃ 1.60 d (1.0) 18.1, CH ₃ 1.62 d (1.0) 11 ' 21.6, CH ₃ 0.91 d (7.0) 21.8, CH ₃ 0.91 d (6.5) 12 ' 19.8, CH ₃ 1.02 d (7.0) 20.3, CH ₃ 1.00 d (6.5) 2'-CH ₃ 12.7, CH ₃ 1.88 d (1.0)

b : ¹ H 600 MHz, ¹³ C 150 MHz.

The chemical structures of the soltanamides A to D analyzed from nuclear magnetic resonance spectral data are shown below.

[Solutin amide A and B]

In the above structural formula, the salt amide A is R ₁ is Cl, and the salt amide B is R ₁ .

[Solutinamide C and D]

In the above structural formula, the salt amide C is R ₃ is H, and the salt amide D is R ₃ is CH ₃ .

5. Saltenamide  Confirmation of anticancer activity of A to D

(Korean Cell Line Bank), the cancer cell line HCT116 (Korean Cell line Bank), the gastric cancer cell line SNU638 (Korean Cell line Bank), the leukemia cell line K562 (Korean Cell line Bank) , Korean hepatocellular carcinoma line SK-HEP1 (Korean Cell line Bank), and breast cancer cell line MDA-MB231 (Korean Cell line Bank).

Specifically, cells were inoculated in a 96-well microplate at a concentration of 3.5 x 10 ³ cells / ml per well of 190 μl. 0.8 μM, 4 μM, 20 μM, or 100 μM of each of the Salten amides A to D was added to each well to which the cells were inoculated, and the cells were incubated in an incubator at 37 ° C. and 5% CO ₂ for 72 hours. Etoposide (Sigma Aldrich) was used as a reference drug. Cells were then fixed by adding 50 [mu] l of 50% (v / v) trichloroacetic acid solution (Sigma Aldrich) per well to each well and incubating at 4 [deg.] C for 30 minutes. The fixed cells were washed 5 times with water and dried in air. Subsequently, cells were added with 80 μl of 0.4% (w / v) Sulforhodamine B (SRB) aqueous solution (Sigma Aldrich) containing 1% (v / v) acetic acid (DUKSAN) Lt; / RTI > for 1 hour to stain the cells. The stained cells were washed with water and dried. Cells were lysed by adding 200 μl of 10 mM Tris (pH 10.0) (Sigma Aldrich) to each well, and the number of living cells was measured by measuring the absorbance at 515 nm. 50% inhibitory concentration of the compound on the cancer cell line, i.e. IC ₅₀ (inhibitory concentration 50%), was calculated and the results are shown in Table 3 below.

50% cell growth inhibitory concentration (IC ₅₀ , uM) of the sul- tonamides A to D, sample A549
(Lung cancer) HCT116
(Colon cancer) SNU638
(Gastric cancer) K562
(leukemia) SK - HEP1
(Liver cancer) MDA - MB231
(Breast cancer) Solutin Amide A 36 0.96 0.96 8.9 2.7 8.9 Solutinamide B 82 55 27 > 100 56 27 Solutinamide C > 100 > 100 > 100 > 100 > 100 > 100 Solutinamide D > 100 34 15 59 32 92 Etoposide 0.80 1.9 0.50 3.1 1.1 11

As shown in Table 3, the IC ₅₀ values of the Soltan amide A for lung cancer cell A549, colon cancer cell HCT116, stomach cancer cell SNU638, leukemia cell K562, liver cancer cell SK-HEP1, and breast cancer cell MDA-MB231 were 36 μM and 0.96 μM, 0.96 μM, 8.9 μM, 2.7 μM, and 8.9 μM, respectively, and significantly inhibited the growth of these cancer cells.

Korea Biotechnology Research Institute KCTC12832BP 20150604

<110> SEOUL NATIONAL UNIVERSITY R & DB Foundation <120> Salternamide compound, preparing method and use thereof <130> PN109901 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 676 <212> DNA <213> Artificial Sequence <220> <223> 16S rDNA of Streptomyces sp. HK10 <400> 1 ttcgggttgt aaacctcttt cagcagggaa gaagcggaag tgacggtacc tgcagaagaa 60 gcaccggcta actacgtgcc agcagccgcg gtaatacgta gggtgcgagc gttgtccgga 120 attattgggc gtaaagagct cgtaggcggc ctgtcgcgtc ggatgtgaaa gcccggggct 180 taaccccggg tcggcattcg atacgggcag gctagagttc ggtaggggag atcggaattc 240 ctggtgtagc ggtgaaatgc gcagatatca ggaggaacac cggtggcgaa ggcggatctc 300 tgggccgata ctgacgctga ggagcgaaag cgtggggagc gaacaggatt agataccctg 360 gtagtccacg ccgtaaacgt tgggcactag gtgtgggcgg cattccacgt cgtccgtgcc 420 gcagctaacg cattaagtgc cccgcctggg gagtacggcc gcaaggctaa aactcaaagg 480 aattgacggg ggcccgcaca agcggcggag catgtggctt aattcgacgc aacgcgaaga 540 accttaccaa ggcttgacat acaccggaaa cggccagaga tggtcgcccc cttgtggtcg 600 gtgtacaggt ggtgcatggc tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg 660 caacgagcgc aactcc 676

Claims (16)

A compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
[Formula 1]

In Equation (1)
R ₁ is a halogen group,
R ₂ is hydrogen, a halogen group, a hydroxy group, an unsubstituted C ₁ -C ₂₀ alkyl group, or a combination thereof,
R ₃ is hydrogen or an unsubstituted C ₁ -C ₂₀ alkyl group,
Provided that R ₁ and R ₂ are not fused to each other to form a ring. delete 2. The compound according to claim 1, wherein the halogen group is -Cl, -F, -Br, or -I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. The compound according to claim 1, wherein the compound represented by Formula 1 is represented by Formula 2, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
[Formula 2]

In the above formula 2,
R ₁ is a halogen group. 5. The compound according to claim 4, wherein the formula 2 is Formula 3, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
[Formula 3]

. delete 5. The compound according to claim 4, wherein the halogen group is -Cl, -F, -Br, or -I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. delete delete delete delete Streptomyces strain HK10 strain (accession number: KCTC12832BP) producing the compound of claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. Culturing Streptomyces sp. Strain HK10 (accession number: KCTC12832BP); And
A method for producing a compound of claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, which comprises the step of isolating the compound of claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof from a culture in which said strain has been cultured How to. A pharmaceutical composition for preventing or treating cancer, comprising the compound of claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. 15. The pharmaceutical composition according to claim 14, wherein the cancer is lung cancer, colon cancer, gastric cancer, leukemia, liver cancer, or breast cancer. A method for preventing or treating cancer, comprising administering to a mammal other than a human a pharmaceutical composition for preventing or treating cancer comprising the compound of claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

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