KR20230142242A - Novel peptide-polyketide compound, or optical isomer thereof, or pharmaceutically acceptable salts thereof, uses thereof, and preparation method thereof - Google Patents

Abstract

The present invention provides a compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition containing the same for inhibiting cancer metastasis or a pharmaceutical composition for preventing or treating cancer diseases.

Description

Novel peptide-polyketide compound, or optical isomer thereof, or pharmaceutically acceptable salts thereof, uses thereof, and preparation method thereof there}

The present invention relates to a novel peptide-polyketide compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, and more specifically, to a composition for preventing, improving, or treating cancer diseases by having cancer cell metastasis inhibitory activity.

Cancer is generally a disease in which cells do not differentiate normally and continue to divide due to abnormalities in the cell cycle that makes up human tissue. It occurs through three stages: initiation, promotion, and progression. . The incidence of cancer is increasing every year due to various social and environmental factors, and cancer is a representative disease that must be taken very seriously. From basic research at the cellular level to comprehensive research on anticancer treatment has been promoted worldwide. come. It is known that cancer is caused by mutations in normal cell genes or cancer suppressor genes caused by carcinogens contained in the environment or food, and that these cells proliferate abnormally and form cancerous tissue while continuously stimulated by carcinogens. The cause of cancer is still not clearly known.

Among them, breast cancer is the most common malignant disease in women. In Korea, it ranked fifth among all cancers after stomach, lung, liver, and colon cancer, accounting for 6.5% (5,444 people) of all cancers in 2005. According to the Ministry of Health and Welfare's Central Cancer Registry Report, breast cancer was the most malignant cancer in women in 2002. It ranked first, accounting for 16.8% of tumors. In the case of breast cancer, early detection is more important than other cancers because it is difficult to cure once cancer cells invade surrounding tissues or metastasize to the lymph nodes.

Cancer is divided into benign tumors and malignant tumors. Benign tumors have a relatively slow growth rate and do not metastasize from the original tumor site to other tissues, whereas malignant tumors do not occur. It has the characteristic of leaving the primary site and infiltrating other tissues and growing rapidly, threatening life and becoming a very important cause of death, and 90% of cancer-related deaths are due to cancer metastasis.

Synthetic anticancer drugs developed to treat these cancers have the function of inhibiting cell proliferation or inducing cell death by inhibiting the metabolism of cancer cells, or inhibiting the DNA replication process by unwinding the double helix structure of DNA during cell division. It has the function of inhibiting the proliferation of cancer cells or eliminating cancer cells by inducing cell death by inhibiting cell division. However, in the case of synthetic anticancer drugs, they attack normal cells, resulting in side effects such as hair loss, diarrhea, and fever. Therefore, targeted anti-cancer drugs are being developed that reduce side effects and improve anti-cancer efficacy. They enhance the effectiveness of synthetic anti-cancer drugs by targeting mutant genes involved in cancer development rather than normal cells. However, resistance mechanisms for most targeted anticancer drugs develop within 2 to 3 years, and the targeted anticancer drugs no longer show therapeutic effects.

In addition, maintaining immunity and homeostasis is the most important issue in health care, and when immunity is lowered, it can easily turn into disease when exposed to a harmful environment or when infected. Continuous treatment with anticancer drugs and antibiotics to treat diseases weakens the body's immunity and slows down the disease relief and treatment effects caused by the drugs, which can lead to secondary diseases.

Bioactive substances derived from microorganisms have largely been the source of antibacterial, antifungal, and anticancer drugs, and have been developed as new drugs for the treatment of various diseases or have served as a template for new drug development. Representative examples of antibiotics derived from microorganisms include amphotericin, erythromycin, streptomycin, tetracycline, and vancomycin. Additionally, in 2003, daptomycin, isolated from the actinomycete Streptomyces, was approved by the U.S. Food and Drug Administration (FDA) as a next-generation antibiotic. In addition, lipstatin, discovered in the bacterium Streptomyces toxytricini, is a medicine for dieting and calorie suppression, and is sold as a lipase inhibitor called Orlistat after some chemical modification. It is becoming. As such, research on bioactive substances derived from bacteria, especially actinomycetes, is very important in pharmaceutical and drug development. It is necessary to select microorganisms that are structurally different from existing substances and produce beneficial bioactive substances, and to explore and discover new compounds they produce.

However, although researchers have made efforts to develop strains with anticancer effects, to date, there has been no case in which strains with excellent anticancer effects have been developed into pharmaceuticals.

Accordingly, the present inventors completed the present invention by discovering that novel peptide-polyketide compounds derived from soil actinomycetes and their derivatives have anticancer, antitumor, or cancer metastasis inhibitory effects, thereby treating and improving cancer diseases.

The purpose of the present invention is to provide a novel peptide-polyketide compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

Additionally, the present invention aims to provide a method for producing a novel peptide-polyketide compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

Additionally, the present invention aims to provide a pharmaceutical composition for preventing or treating cancer diseases, comprising a novel peptide-polyketide compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

In addition, the present invention aims to provide a pharmaceutical composition for inhibiting cancer metastasis comprising a novel peptide-polyketide compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

In addition, the present invention aims to provide a food composition for preventing or improving cancer disease, comprising a novel peptide-polyketide compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

In addition, the present invention relates to a cancer disease comprising the step of administering a therapeutically effective amount of a novel peptide-polyketide compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof to a subject in need thereof. Provides treatment methods.

Another object of the present invention is to provide the use of a novel peptide-polyketide compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof for the production of a drug for treating cancer diseases.

Additionally, the present invention aims to provide a new Streptomyces sp. strain 13F051 (accession number: KCTC 14889BP).

The terms used in this application are only used to describe specific embodiments and are not intended to limit the invention. Singular expressions include plural expressions unless the context clearly dictates otherwise. In this application, terms such as “comprise” or “have” are intended to designate the presence of the features, steps, structures, or combinations thereof described above in the name, but are intended to indicate the presence of one or more other features, steps, structures, or these. It should be understood that the existence or addition of combinations is not excluded in advance.

Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as generally understood by a person of ordinary skill in the technical field to which the present invention pertains. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related technology, and unless explicitly defined in the present application, should not be interpreted in an ideal or excessively formal sense. No.

In order to solve the above problem, the present inventors isolated and identified a peptide-polyketide compound represented by the following formula (1) from a fraction of the culture medium of Streptomyces sp. 13F051 strain (accession number: KCTC 14889BP), and its optical isomer. The present invention was completed by discovering that a pharmaceutically acceptable salt thereof inhibits cancer cell metastatic activity, has anti-cancer and anti-tumor effects, and is effective in preventing, improving, and treating cancer diseases.

Hereinafter, the new peptide-polyketide compound of the present invention, its production method, and its use will be described in detail.

Novel peptide-polyketide compounds, optical isomers thereof or pharmaceutically acceptable salts thereof

In one aspect of the present invention, a compound represented by the following formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof is provided.

[Formula 1]

.

In the present invention, the compound represented by Formula 1, its optical isomer, or its pharmaceutically acceptable salt may be derived from Streptomyces sp. 13F051 strain (Accession number: KCTC 14889BP), The strain may have the gene sequence of SEQ ID NO: 1 as follows.

[SEQ ID NO: 1]

TCATGGCTCAGGACGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGATGAAGCCTTTCGGGGTGGATTAGTGGCGAACGGGTGAGTAACACGTGGGCAATCTGCCCTTCACTCTGGGACAAGCCCTGGAAACGGGGTCTAATACCGGATATGACACACGACCGCATGGTCTGTGTGTGTGAAAGCTCCGGCGGTGAAGGATGAGCCCGCGGCCTATCAGCTTGTTGGTGGGGTGATGGCCTAC CAAGGCGACGACGGGTAGCCGGCCTGAGAGGGCGACCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGAAAGCCTGATGCAGCGACGCCGCGTGAGGGATGACGGCCTTCGGGTTGTAAACCTCTTTCAGCAGGGAAGAAGCGAGAGTGACGGTACCTGCAGAAGAAGCGCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGC GCAAGCGTTGTCCGGAATTATTGGGCGTAAAGAGCTCGTAGGCGGCTTGTCACGTCGGATGTGAAAGCCCGGGGCTTAACCCCGGGTCTGCATTCGATACGGGCAGGCTAGAGTTCGGTAGGGGAGATCGGAATTCCTGGTGTAGCGGTGAAATGCGCAGATATCAGGAGGAACACCGGTGGCGAAGGCGGATCTCTGGGCCGATACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCT GGTAGTCCACGCCGTAAACGTTGGGAACTAGGTGTGGGCGACATTCCACGTCGTCCGTGCCGCAGCTAACGCATTAAGTTCCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCAGCGGAGCATGTGGCTTAATTCGACGCAACGCGAAGAACCTTACCAAGGCTTGACATACACCGGAAAACCCTGGAGACAGGGTCCCCCTTGTGGTCGGTGTACAGGTGGTGCATGG CTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTTCTGTGTTGCCAGCATGCCCTTCGGGGTGATGGGGACTCACAGGAGACTGCCGGGGTCAACTCGGAGGAAGGTGGGGACGACGTCAAGTCATCATGCCCCTTATGTCTTGGGGCTGCACACGTGCTACAATGGCCGGTACAATGAGCTGCGATACCGCGAGGTGGAGCGAATCTCAAAAAGCCGGTCTCAGTTCGG ATTGGGGTCTGCAACTCGACCCCATGAAGTCGGAGTTGCTAGTAATCGCAGATCAGCATTGCTGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACGTCACGAAAGTCGGTAACACCCGAAGCCGGTGGCCCAACCCCTTGTGGGAGGGAATCGTCGAAGGTGGGACTGGCGATTGGGAACGAAGTCGTAACAAGGTA.

The novel peptide-polyketide compound represented by Formula 1 was named Ulleungdolin.

The compound represented by Formula 1 of the present invention includes not only its salt but also optical isomers that can be prepared therefrom by conventional methods.

In the present invention, an acid addition salt formed from a free acid is useful as the “salt” of the compound represented by Formula 1. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, etc., aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes. It is obtained from non-toxic organic acids such as dioate, aromatic acids, aliphatic and aromatic sulfonic acids, and organic acids such as acetate, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, and fumaric acid. These salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, iodide, and fluoride. , acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, Fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methyl benzoate, dinitro benzoate, hydroxybenzoate, methoxybenzoate, Phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, maleate, Includes tartrate, methanesulfonate, propane sulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate, etc.

In addition, the acid addition salt can be prepared by conventional methods, for example, by dissolving the compound represented by Formula 1 in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc. and adding an organic acid or inorganic acid. It can be prepared by filtering and drying the precipitate, or by distilling the solvent and excess acid under reduced pressure, drying it, and crystallizing it in an organic solvent.

Additionally, metal salts can be made using bases. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically appropriate to prepare sodium, potassium, or calcium salts as metal salts. Additionally, the corresponding salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (e.g., silver nitrate).

In the present invention, the term “optical isomer” refers to two stereoisomers of a compound that have non-overlapping mirror images, and diastereomers refer to stereoisomers that have two or more asymmetric centers and whose molecules are not mirror images of each other.

“Pharmaceutically acceptable salt” of the present invention refers to salts commonly used in the pharmaceutical industry, for example, inorganic ionic salts made of calcium, potassium, sodium and magnesium, hydrochloric acid, nitric acid, phosphoric acid, and bromic acid. , inorganic acid salts prepared from iodic acid, perchloric acid, sulfuric acid, etc.; Acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid. Organic acid salts made from acids, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, etc.; Sulfonic acid salts made from methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, etc.; Amino acid salts made from glycine, arginine, lysine, etc.; and amine salts prepared from trimethylamine, triethylamine, ammonia, pyridine, picoline, etc., but the types of salts meant in the present invention are not limited by these salts listed.

Specifically, the peptide-polyketide compound represented by Formula 1, its optical isomer, or its pharmaceutically acceptable salt is a culture fraction of Streptomyces sp. 13F051 strain (accession number: KCTC 14889BP) It was confirmed through examples that it is effective in suppressing the metastatic activity of cancer cells, and it can be seen that it is effective in treating cancer diseases (FIG. 4). Therefore, the peptide-polyketide compound represented by Formula 1 of the present invention can be used as an active ingredient in a drug for preventing, improving, or treating cancer diseases.

Method for producing novel peptide-polyketide compounds, optical isomers thereof, or pharmaceutically acceptable salts thereof

In one aspect of the present invention, in order to obtain the compound of Formula 1, the culture fraction of Streptomyces sp. 13F051 strain (accession number: KCTC 14889BP) is prepared by a production method comprising the following steps. Can be manufactured, but is not limited to:

1) isolating and culturing the Streptomyces sp. 13F051 (accession number: KCTC 14889BP) strain to obtain a strain culture;

2) extracting the strain culture obtained in step 1) with an organic solvent to obtain an extract; and

3) Fractionating the extract obtained in step 2) using an organic solvent.

In the present invention, in step 1), the strain can be cultured in a medium containing a nutrient source that can be used by ordinary microorganisms. As a nutrient source, a known nutrient source conventionally used for culturing actinomycetes can be used. For example, glucose, starch syrup, dextrin, starch, molasses, animal oil, vegetable oil, etc. can be used as carbon sources, and wheat bran, soybean meal, wheat, malt, cottonseed meal, fish meal, corn syrup, meat juice, yeast extract, etc. can be used as nitrogen sources. Ammonium sulfate, sodium nitrate, urea, tryptone, etc. can be used. If necessary, table salt, potassium, magnesium, cobalt, chlorine, phosphoric acid, sulfuric acid and other inorganic salts that promote ion production can be added. As a culture method, shaking culture or static culture is possible under aerobic conditions. The culture temperature varies slightly depending on the conditions when culturing under each of the above conditions, but generally, the culture can be performed at 20°C to 37°C and can also be cultured at 25°C to 30°C, but is not limited thereto. In one embodiment of the present invention, the culture medium may be SY medium containing soluble starch, yeast extract, tryptone, agar, etc.

The organic solvent may be water, C ₁ to C ₄ lower alcohol, hexane, chloroform, methylene chloride, ethyl acetate, acetone, acetonitrile, and combinations thereof.

In step 2), the peptide-polyketide compound according to the present invention may be present not only in the culture medium of the strain but also in the bacterial cell portion. Therefore, an organic solvent is added to the culture medium and the bacterial cells to extract the active ingredients from the culture medium and the bacterial cells, and then the obtained extract is concentrated by a reduced pressure evaporation method. At this time, the organic solvent that can be used may be ethyl acetate, acetone, etc., and may be specifically acetone, but the type of organic solvent used for extraction is not limited thereto.

Additionally, in step 3), the organic solvent may be water, ethanol, methanol, acetonitrile, and a combination thereof, and preferably may be a mixed solvent of water and methanol. Specifically, it can be eluted and fractionated by gradually increasing the methanol concentration using methanol/water (20:80-80:20, v/v) as a mixed solvent.

In the present invention, a fraction of a culture of Streptomyces sp. 13F051 (Accession number: KCTC 14889BP) is fractionated using thin layer chromatography or liquid chromatography techniques to isolate the compound represented by Formula 1. .

In the present invention, the fractionation in step 3) may be performed on the extract obtained in step 2) by column chromatography using a mixed solvent of methanol and water, and then purified by high-performance liquid chromatography.

The column chromatography and high-performance liquid chromatography techniques refer to chromatography techniques in which the mobile phase is a liquid, and are performed in a column filled with a stationary phase or a plane to which the stationary phase is attached. The fraction of the culture of the Streptomyces sp. 13F051 strain (accession number: KCTC 14889BP) of the present invention is not limited as long as it is a solvent fraction. As the mobile phase, organic solvents such as water, hexane, methanol, ethanol, acetonitrile, acetone, chloroform, dichloromethane, and ethyl acetate can be used alone or in combination, and as the stationary phase, silica gel and Diaion HP-20 can be used. , RP-18 or Sephadex LH-20 can be used, but are not limited to these.

In one embodiment of the present invention, the peptide-polyketide compound of the present invention was extracted and isolated from the terrestrial soil actinomycete Streptomyces sp. 13F051 (Accession number: KCTC 14889BP) strain.

Pharmaceutical composition or food composition comprising a novel peptide-polyketide compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof

The present invention provides a pharmaceutical composition for preventing or treating cancer diseases, comprising the compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

Additionally, the present invention provides a pharmaceutical composition for inhibiting cancer metastasis comprising the compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

The compound according to the present invention can be used to treat cancer diseases by inhibiting the metastatic activity of cancer cells, and in particular, can be used to treat metastatic cancer.

In the present invention, the cancer disease may be a metastatic cancer disease.

In the present invention, the cancer disease includes lung cancer, non-small cell lung cancer (NSCL), bronchoalveolar cell lung cancer, stomach cancer, gastrointestinal cancer, liver cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or eye melanoma, uterine cancer, and ovarian cancer. , rectal cancer, colon cancer, colon cancer, breast cancer, uterine sarcoma, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, esophagus cancer, laryngeal cancer, small intestine cancer, thyroid cancer, parathyroid adenocarcinoma, sarcoma of soft tissue, urethral cancer, penis It may be cancer, prostate cancer, multiple myeloma, chronic or acute leukemia, solid tumor of childhood, lymphoma, bladder cancer, kidney cancer, renal cell carcinoma, renal pelvic carcinoma, axial contractile tumor, brainstem glioma or pituitary adenoma, etc. may be breast cancer, and more specifically, may be metastatic breast cancer.

For administration, the pharmaceutical composition of the present invention may further include one or more pharmaceutically acceptable carriers in addition to the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof. Pharmaceutically acceptable carriers can be saline solution, sterile water, Ringer's solution, buffered saline solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and a mixture of one or more of these ingredients, and if necessary, antioxidants and buffer solutions. Other common additives such as bacteriostatic agents can be added. In addition, diluents, dispersants, surfactants, binders, and lubricants can be additionally added to formulate injectable formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets. Accordingly, the pharmaceutical composition of the present invention may be a patch, solution, pill, capsule, granule, tablet, suppository, etc. These preparations can be manufactured by conventional methods used for formulation in the art or by methods disclosed in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA, and can be formulated into various preparations depending on each disease or ingredient. It can be.

The composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, the term "pharmaceutically effective amount" refers to an amount that is sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment and does not cause side effects, and the effective dose level is determined by the patient's Factors including health status, type and severity of disease, activity of drug, sensitivity to drug, method of administration, time of administration, route of administration and excretion rate, duration of treatment, drugs combined or used simultaneously, and other factors well known in the field of medicine. It can be decided depending on The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. Considering all of the above factors, it is important to administer an amount that can achieve maximum effect with the minimum amount without side effects, and this can be easily determined by a person skilled in the art. The daily dosage of the compound of Formula 1 of the present invention is about 0.01 to 1000 mg/kg, preferably 0.1 to 100 mg/kg, and can be administered once to several times a day.

The term "administration" of the present invention means introducing a predetermined substance into a patient by an appropriate method, and the composition may be administered through any general route as long as it can reach the target tissue. Additionally, the pharmaceutical composition of the present invention may be administered by any device that allows the active substance to move to the target tissue. For example, oral administration, intrathecal administration, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, topical administration, intranasal administration, intrapulmonary administration, intrarectal administration, inner ear administration, intrauterine intrathecal administration, It may be administered by sublingual administration or intracerebrovascular injection, but is not limited thereto.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations contain at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc. Formulated by mixing. Additionally, in addition to simple excipients, lubricants such as magnesium stearate and talc may be used.

Liquid preparations for oral use include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, fragrances, and preservatives. You can.

Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. The basis of suppositories is Wethepsol, Macrogol, and Twin 61. Cacao, laurel, glycerogeratin, etc. can be used. Meanwhile, injectables may contain conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers, preservatives, etc.

The composition may be administered through any general route as long as it can reach the target tissue, but may include subcutaneous injection, intradermal injection, or intravein injection using an osmotic pump. ), can be administered through intraperitoneal injection, or intravitreal injection.

The pharmaceutical composition of the present invention may further include one or more active ingredients exhibiting the same or similar medicinal efficacy in addition to the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof.

In another aspect, the present invention provides a food composition for preventing or improving cancer disease, comprising the compound represented by Formula 1, an optical isomer thereof, or a foodologically acceptable salt thereof.

The composition may include food additives that are foodologically acceptable in addition to the active ingredients.

The term "food supplement" used in the present invention refers to a component that can be added as an auxiliary food additive, and can be appropriately selected and used by a person skilled in the art as it is added to manufacture each type of health functional food. Examples of food supplements include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavors, colorants and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. are included, but the types of food supplements of the present invention are not limited to the above examples.

The food composition of the present invention may include health functional foods. The term “health functional food” used in the present invention refers to food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and pills using raw materials or ingredients with functional properties useful to the human body. Here, ‘functionality’ means controlling nutrients for the structure and function of the human body or obtaining useful effects for health purposes, such as physiological effects. The health functional food of the present invention can be manufactured by a method commonly used in the industry, and can be manufactured by adding raw materials and components commonly added in the industry. Additionally, the formulation of the health functional food can also be manufactured without limitation as long as it is a formulation recognized as a health functional food. The food composition of the present invention can be manufactured in various types of formulations, and unlike general drugs, it is made from food as a raw material and has the advantage of not having side effects that may occur when taking the drug for a long period of time, and is excellent in portability, so the present invention Health functional foods can be consumed as supplements to enhance the effectiveness of anticancer drugs.

The term “prevention” used in the present invention refers to any action that inhibits cancer formation or delays the onset of cancer by administering a composition.

As used herein, the term “improvement” means any action that reduces at least the severity of a parameter, such as a symptom, related to the condition being treated.

In the present invention, “treatment” means any action in which the symptoms of the disease are improved or beneficially changed by administration of the composition.

Uses, methods and use thereof in the manufacture of medicaments for the treatment of cancer diseases

The present invention provides a method of treating cancer disease, comprising administering a therapeutically effective amount of the compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof to a subject in need thereof.

Subjects that require this are mammals, including humans, such as humans, monkeys, cows, horses, dogs, cats, rabbits, rats, and mice.

The term "therapeutically effective amount" as used in the present invention refers to the amount of the novel salt or a pharmaceutical composition containing the same that is effective in preventing or treating a cancer disease, such as the amount of the new salt administered to the subject to be treated, Prevent the occurrence or recurrence of cancer disease, alleviate symptoms, impede direct or indirect pathological consequences, prevent metastasis, reduce the rate of progression, alleviate or temporarily alleviate the condition, or provide prognosis. It may contain all the amounts of the pharmaceutical composition containing the above-mentioned salts that improve. In other words, the therapeutically effective amount can be interpreted as encompassing all doses at which symptoms of cancer disease are improved or completely cured by the pharmaceutical composition.

The method for preventing or treating a cancer disease of the present invention includes not only treating the disease itself before the onset of symptoms, but also inhibiting or avoiding its symptoms, by administering the above-mentioned salt or a pharmaceutical composition containing the same. In the management of disease, the prophylactic or therapeutic dosage of a particular active ingredient will vary depending on the nature and severity of the disease or condition and the route by which the active ingredient is administered. Dosage and frequency of dosage will vary depending on the age, weight, and response of the individual patient. A suitable dosage regimen can be easily selected by one of ordinary skill in the art taking these factors into account. In addition, the method of treating the cancer disease of the pharmaceutical composition of the present invention may further include administration of a therapeutically effective amount of an additional active agent helpful in treating the disease along with the above-mentioned salt, and the additional active agent is the present invention. It may exhibit a synergistic or auxiliary effect together with the above-mentioned salt, which is an active ingredient according to the invention.

The present invention also seeks to provide the use of the compound represented by Formula 1, its optical isomer, or its pharmaceutically acceptable salt for the production of a drug for treating cancer diseases. The above-mentioned salts for the preparation of drugs can be mixed with acceptable auxiliaries, diluents, carriers, etc., and can be prepared as a complex preparation with other active agents to have a synergistic effect of the active ingredients.

The present invention also provides a pharmaceutical composition for use in the prevention or treatment of cancer disease, comprising the compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

Matters mentioned in the uses, compositions, and treatment methods of the present invention apply equally unless they contradict each other.

The novel peptide-polyketide compound according to the present invention has cancer cell metastasis inhibitory activity and has anti-cancer and anti-tumor effects, so it can be used in the treatment of cancer diseases.

In addition, the compound of the present invention is a natural product that can minimize side effects, and because it uses culturable actinomycetes, it is easy to extract and purify, making it very advantageous in terms of economic efficiency.

Figure 1 shows a process diagram (a) for isolating a compound from a culture fraction of the Streptomyces sp. 13F051 (accession number: KCTC 14889BP) strain and the structure (b) of the novel peptide-polyketide compound of the present invention. ) is shown.
Figure 2 shows the HPLC profile of the peptide-polyketide compound of Formula 1 of the present invention.
Figure 3 shows the ¹ H-NMR (a) and ¹³ C-NMR (b) spectra of the peptide-polyketide compound of Formula 1 of the present invention.
Figure 4 shows the results of analyzing the effect on the migration of the metastatic breast cancer cell line MDA-MB-231 according to treatment with the peptide-polyketide compound of Formula 1 of the present invention.

Hereinafter, it will be described in detail through examples.

However, the following examples only illustrate the present invention, and the content of the present invention is not limited to the following examples.

<Example 1> Streptomyces genus ( Streptomyces sp.)　13F051 strain isolation, deposit and culture

(1) Streptomyces genus ( Streptomyces sp.)　13F051 strain isolation and identification

Soil samples were collected from Ulleungdo Island, South Korea. Isolated strain 13F051 was most similar to Streptomyces sioyaensis NRRL B-5408 (99%, 1442/1449) , Streptomyces inhibens NEAU-D10 (99%, 1442/1449), and Streptomyces auratus NRRL 8097 (99%, 1439/1449). It was found to have a 16S rRNA gene sequence (GenBank accession number: MW774247).

Therefore, strain 13F051 was transformed into Streptomyces sp. It was named 13F051 and used in subsequent culture experiments.

The Streptomyces sp. The strain named 13F051 was deposited at the Korea Microbial Resources Center on March 3, 2022 with the accession number KCTC14889BP.

(2) Streptomyces genus ( Streptomyces sp.) Culture of 13F051 strain

To cultivate this actinomycete strain, Streptomyces sp. was maintained on SY agar medium containing 10.0 g soluble starch, 1.0 g yeast extract, 1.0 g tryptone, and 18.0 g agar per 1 L of distilled water. ) 13F051 per 1 L of distilled water, 30.0 g dextrose, 5.0 g yeast extract, 0.05 g adenine, 0.05 g L-histidine, 0.05 g L-leucine, 0.05 g L-lysine HCl, 0.05 g uracil, After streaking in the center of a 6 cm YES agar plate containing 20.0 g agar, the cells were cultured at 25°C for 21 days under stationary conditions.

<Example 2> Streptomyces genus ( Streptomyces sp.) Isolation and purification of peptide-polyketide compounds from strain 13F051

Streptomyces sp. 13F051 strain contained in 100 Petri dishes cultured in Example 1 The culture medium was soaked in acetone three times and then evaporated to remove the solvent to obtain a crude extract (acetone extract). The obtained crude extract (2.5 g) was treated with a graded solvent system of MeOH-H ₂ O (1 L for each step, stepwise increasing concentration of methanol from 20:80, 40:60, 60:40, 80:20 to 100:0). ) was fractionated by reverse-phase C18 flash column chromatography.

Fraction 4 (MeOH-H ₂ O=80:20) was subjected to reverse-phase C18 preparative HPLC (flow rate 15 mL/min) using a graded solvent system of CH ₃ CN-H ₂ O (25:75 to 60:40). After purification, compound (1) showing UV absorption peaks at 290 and 350 nm was obtained (10.3 mg, t _R 36.2 min).

Ulleungdolin (1): Brown amorphous powder; [α]25 D -50.0 (c 0.04, MeOH); UV (MeOH) l _max (log e ) 231 (2.20), 251 (3.32), 287 (3.04) nm.

<Example 3> Streptomyces genus ( Streptomyces sp.) Structural analysis of peptide-polyketide compounds from strain 13F051

The molecular weight and molecular formula of the peptide-polyketide compound according to the present invention prepared from the culture medium of the Streptomyces sp. 13F051 strain were determined using electrospray ionization mass spectrometer (ESIMS). . In addition, ¹ H NMR and ¹³ C NMR spectra were obtained through nuclear magnetic resonance (NMR) analysis (Bruker AVANCE HD 800 NMR spectrometer), and the molecular structure of the compound was determined.

Compound (1), a substance isolated from the culture medium of the Streptomyces sp. strain 13F051, was identified as a novel peptide-polyketide compound of Formula 1. The results are shown in Table 1 below, and ^{the 1} H-NMR and ¹³ C-NMR analysis results are shown in Figure 3.

<Experimental Example 1> Analysis of cancer cell metastasis inhibition activity of Formula 1

To confirm the effect of the compound of Formula 1 on breast cancer cell migration, human breast adenoma MDA-MB-231 cells, a metastatic breast cancer cell line, were analyzed by wound migration assay.

Specifically, the MDA-MB-231 cell line was seeded in a 24-well plate at 9 x 10 ⁴ cells per well, and after 18 h, the center of the well was scratched with a 200 μl tip. Treated with 0 μM, 10 μM, and 25 μM, 24 h later, stained with 0.2% crystal violet reagent and observed under a microscope, the results are shown in Figure 4.

As can be seen in Figure 4, MDA-MB-231 treated with the compound of the present invention showed low mobility, and it was confirmed that the migration of cancer cells was significantly inhibited by treatment with Compound 1 of the present invention.

Above, the present invention has been described by way of example, and those skilled in the art will be able to make various modifications without departing from the essential characteristics of the present invention. Accordingly, the embodiments disclosed in this specification are for illustrative purposes rather than limiting the present invention, and the spirit and scope of the present invention are not limited by these embodiments. The scope of protection of the present invention should be interpreted in accordance with the claims below, and all technologies within the equivalent scope should be interpreted as being included in the scope of rights of the present invention.

Korea Research Institute of Bioscience and Biotechnology KCTC14889BP 20220303

<110> Korea Research Institute of Bioscience and Biotechnology <120> Novel peptide-polyketide compound, or optical isomer thereof, or pharmaceutically acceptable salts thereof, uses thereof, and preparation method <130> P22034-KRIBB <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1476 <212> DNA <213> Artificial Sequence <220> <223> KCTC 14889BP <400> 1 tcatggctca ggacgaacgc tggcggcgtg cttaacacat gcaagtcgaa cgatgaagcc 60 tttcggggtg gattagtggc gaacgggtga gtaacacgtg ggcaatctgc ccttcactct 120 gggacaagcc ctggaaacgg ggtctaatac cggatatgac acacgaccgc atggtctgtg 180 tgtggaaagc tccggcggtg aaggatgagc ccgcggccta tcagcttgtt ggtggggtga 240 tggcctacca aggcgacgac gggtagccgg cctgagaggg cgaccggcca cactgggact 300 gagacacggc ccagactcct acgggaggca gcagtgggga atattgcaca atgggcgaaa 360 gcctgatgca gcgacgccgc gtgagggatg acggccttcg ggttgtaaac ctctttcagc 420 agggaagaag cgagagtgac ggtacctgca gaagaagcgc cggctaacta cgtgccagca 480 gccgcggtaa tacgtagggc gcaagcgttg tccggaatta ttgggcgtaa agagctcgta 540 ggcggcttgt cacgtcggat gtgaaagccc ggggcttaac cccgggtctg cattcgatac 600 gggcaggcta gagttcggta ggggagatcg gaattcctgg tgtagcggtg aaatgcgcag 660 atatcaggag gaacaccggt ggcgaaggcg gatctctggg ccgatactga cgctgaggag 720 cgaaagcgtg gggagcgaac aggattagat accctggtag tccacgccgt aaacgttggg 780 aactaggtgt gggcgacatt ccacgtcgtc cgtgccgcag ctaacgcatt aagttccccg 840 cctggggagt acggccgcaa ggctaaaact caaaggaatt gacggggggcc cgcacaagca 900 gcggagcatg tggcttaatt cgacgcaacg cgaagaacct taccaaggct tgacatacac 960 cggaaaaaccc tggagacagg gtcccccttg tggtcggtgt acaggtggtg catggctgtc 1020 gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac gagcgcaacc cttgttctgt 1080 gttgccagca tgcccttcgg ggtgatgggg actcacagga gactgccggg gtcaactcgg 1140 aggaaggtgg ggacgacgtc aagtcatcat gccccttatg tcttgggctg cacacgtgct 1200 acaatggccg gtacaatgag ctgcgatacc gcgaggtgga gcgaatctca aaaagccggt 1260 ctcagttcgg attggggtct gcaactcgac cccatgaagt cggagttgct agtaatcgca 1320 gatcagcatt gctgcggtga atacgttccc gggccttgta cacaccgccc gtcacgtcac 1380 gaaagtcggt aacacccgaa gccggtggcc caaccccttg tgggagggaa tcgtcgaagg 1440 tgggactggc gattgggacg aagtcgtaac aaggta 1476

Claims (11)

A compound represented by the following formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof:
[Formula 1]
. The method of claim 1, wherein the compound, its optical isomer, or pharmaceutically acceptable salt thereof is isolated from Streptomyces sp. 13F051 strain (accession number: KCTC 14889BP). A pharmaceutically acceptable salt thereof. The compound, its optical isomer or its pharmaceutically acceptable salt according to claim 1, wherein the compound, its optical isomer or its pharmaceutically acceptable salt has cancer cell metastasis inhibitory activity. A pharmaceutical composition for inhibiting cancer metastasis comprising the compound according to any one of claims 1 to 3, an optical isomer thereof, or a pharmaceutically acceptable salt thereof. A pharmaceutical composition for preventing or treating cancer, comprising the compound according to any one of claims 1 to 3, an optical isomer thereof, or a pharmaceutically acceptable salt thereof. The pharmaceutical composition according to claim 5, wherein the cancer is metastatic cancer. The method of claim 5, wherein the cancer disease is lung cancer, non-small cell lung cancer (NSCL), bronchoalveolar cell lung cancer, stomach cancer, gastrointestinal cancer, liver cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or eye melanoma, uterine cancer, and ovarian cancer. Cancer, rectal cancer, colon cancer, colon cancer, breast cancer, uterine sarcoma, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, esophageal cancer, laryngeal cancer, small intestine cancer, thyroid cancer, parathyroid adenocarcinoma, sarcoma of soft tissue, urethral cancer, One selected from the group consisting of penile cancer, prostate cancer, multiple myeloma, chronic or acute leukemia, solid tumor of childhood, lymphoma, bladder cancer, kidney cancer, renal cell carcinoma, renal pelvic carcinoma, axial contractile tumor, brainstem glioma, or pituitary adenoma. The pharmaceutical composition as described above. The pharmaceutical composition according to claim 5, wherein the cancer disease is breast cancer. The pharmaceutical composition according to claim 8, wherein the breast cancer is metastatic breast cancer. A food composition for preventing or improving cancer disease, comprising the compound according to any one of claims 1 to 3, an optical isomer thereof, or a pharmaceutically acceptable salt thereof. Streptomyces genus ( Streptomyces sp.) KCTC 14889BP.