KR102553669B1 - Piperidine derivative compounds that inhibit Gli activity and their use in cancer treatment - Google Patents

Abstract

It relates to compounds that inhibit Gli activity and their use in cancer treatment. According to the compound according to one aspect, it not only has a cancer cell-specific killing effect by inhibiting Gli, but also has an effect of suppressing cancer metastasis by inhibiting the migration and invasion of cancer cells, making it a pharmaceutical composition for preventing or treating cancer. There are effects that can be useful.

Description

Piperidine derivative compounds that inhibit Gli activity and their use in cancer treatment {Piperidine derivative compounds that inhibit Gli activity and their use in cancer treatment}

It relates to compounds that inhibit Gli activity and their use in cancer treatment.

Cancer is one of the most common causes of death worldwide, accounting for about 12% of all deaths. In particular, lung cancer is on the rise due to the aging of the population, an increase in the smoking population, and air pollution. Dietary habits have become westernized and high-fat diets have become common, and colorectal and breast cancers have rapidly increased in environmental pollutants and increased alcohol consumption. , prostate cancer, etc. are continuously increasing. In this situation, there is an urgent need to create an anti-cancer substance that can contribute to the improvement of human health, the improvement of healthy life quality, and the promotion of human health by enabling early prevention and treatment of cancer.

Chemotherapy, which is a representative anticancer therapy, is currently used as the most effective treatment for cancer, either alone or in combination with other therapies such as radiation therapy. However, the efficacy of a cancer treatment drug in chemotherapy depends on its ability to kill cancer cells, but there is a problem that when the drug is used, it can act on normal cells as well as cancer cells.

Therefore, there is a need for an anticancer agent that specifically targets cancer cells and has a high therapeutic effect.

is to provide:

[Formula 1]

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Another aspect is to provide a pharmaceutical composition for inhibiting cancer metastasis comprising a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 1]

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.

Another aspect is to provide a health functional food for preventing or improving cancer comprising a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 1]

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.

Another aspect is to provide a method for preventing or treating cancer comprising administering a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof to a subject in need thereof:

[Formula 1]

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.

Another aspect is to provide a use of a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof for the preparation of a drug for preventing or treating cancer:

[Formula 1]

.

.

One aspect provides a pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 1]

.

.

The compound represented by Formula 1 may be named 1-(3-(difluoromethoxy)phenyl)-2-((1-methylpiperidin-4-yl)amino)ethanol, and is referred to as compound #168 in the following examples. .

In the present specification, the term "Gli" is the last sub-signal of Hedgehog signaling, and is known to act as a transcription factor in the nucleus and regulate various genes related to cancer development. Among them, it reduces the expression of p21, known as a cancer suppressor gene, and increases the expression of cyclin D1, known as an oncogene.

In one embodiment, the compound may inhibit Gli (glioma-associated oncogene homolog) gene or Gli protein activity. The Gli may be one or more selected from the group consisting of Gli1, Gli2 and Gli3. As the compound inhibits the expression of Gli, the expression of p21, a subgene, can be increased and the expression of Cyclin D1 can be decreased. The cancer may be a cancer in which Gli is expressed, and thus, the compound has an effect of preventing or treating cancer.

As used herein, the term "compound of the present invention" is meant to include the compound of Formula 1 as well as clathrates, hydrates, solvates, isomers or polymorphs thereof.

The compounds described herein may contain one or more asymmetric centers and thus may exist in various isomeric forms, eg enantiomers and/or diastereomers. For example, the compounds described herein may be in the form of individual enantiomers, diastereomers, or geometric isomers, or may be in the form of mixtures of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. . Isomers can be isolated from mixtures by methods known to those skilled in the art including chiral high pressure liquid chromatography (HPLC) and formation and crystallization of chiral salts; Alternatively, the desired isomer may be prepared by asymmetric synthesis.

As used herein, the term "pharmaceutically acceptable" means exhibiting non-toxic properties to cells or humans exposed to the composition.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt prepared using a specific compound according to one aspect and a relatively non-toxic acid or base. When the compounds contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include salts of sodium, potassium, calcium, ammonium, organic amines, or magnesium or similar salts. When the compounds contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of an acid in neat solution or in a suitable inert solvent. Pharmaceutically acceptable acid addition salts include salts of inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, hydrogen carbonate ion, phosphoric acid, hydrogen phosphate ion, dihydrogen phosphate ion, sulfuric acid, hydrogen sulfate ion, hydroiodic acid or phosphorous acid; and salts of organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, and methanesulfonic acid. and salts of amino acids (eg, arginine) and salts of organic acids such as glucuronic acid.

The pharmaceutically acceptable salt can be synthesized from a parent compound containing an acidic or basic moiety by conventional chemical methods. Generally, these salts are prepared by reacting the free acid or base form of these compounds with an appropriate stoichiometric amount of the base or acid, either in water or in an organic solvent or in a mixture of the two. In general, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred.

In one embodiment, the cancer may be hematological cancer or solid cancer.

The blood cancer may be any one or more selected from the group consisting of leukemia, lymphoma, and myeloma, such as multiple myeloma (MM); active multiple myeloma; asymptomatic multiple myeloma; plasmacytoma; Solitary Plasmacytoma of Bone; extramedullary plasmacytoma; light chain myeloma; nonsecretory myeloma; immunoglobulin G (IgG) myeloma; immunoglobulin A (IgA) myeloma; immunoglobulin M (IgM) myeloma; immunoglobulin D (IgD) myeloma; immunoglobulin E (IgE) myeloma; and Diploid Multiple Myeloma; non-hyperdiploid multiple myeloma; Hodgkin's Lymphoma; non-Hodgkin's lymphoma; acute lymphoblastic leukemia; acute myelogenous leukemia; essential thrombocytosis; polycythemia; primary myelofibrosis; systemic mastocytosis; chronic myelogenous leukemia; chronic neutrophilic leukemia; chronic eosinophilic leukemia; refractory anemia with iron ball annulare; refractory cytopenia with multiple systemic dysplasia; Refractory anemia with excessive blasts type 1; Refractory anemia with excessive blasts type 2; isolated del (5q) myelodysplastic syndrome (MDS); MDS Unclassified; chronic myelomonocytic leukemia (CML); atypical chronic myelogenous leukemia; juvenile myelomonocytic leukemia; myeloproliferative/myelodysplastic syndrome-unclassified; B lymphoblastic leukemia/lymphoma; T lymphoblastic leukemia/lymphoma; diffuse large B cell lymphoma; Primary Central Nervous System Lymphoma; Primary Mediastinal B-Cell Lymphoma; Burkitt's Lymphoma/Leukemia; follicular lymphoma; chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma; B cell prolymphocytic leukemia; lymphoplasmacytic lymphoma/Waldenstrom's macroglobulinemia; mantle cell lymphoma; marginal zone lymphoma; post-transplant lymphoproliferative disorders; HIV-related lymphoma; primary effusion lymphoma; Intravascular Large B Cell Lymphoma; Primary Cutaneous Primary Cutaneous B Cell Lymphoma; hairy cell leukemia; monoclonal gammopathy of unknown significance; Anaplastic Giant Cell Lymphoma; angioimmunoblastic T-cell lymphoma; Hepatosplenic T-cell lymphoma, B-cell lymphoma, reticuloendotheliosis, delusionosis, mucosal-associated lymphoid tissue lymphoma, B-cell chronic lymphocytic leukemia, Waldenstrom's macroglobulinemia, lymphomatoid granulomatosis, nodular lymphocyte-predominant Hodgkin's lymphoma, plasma cell leukemia , acute erythroleukemia and erythrocytic leukemia, acute erythrocytic myelosis, acute erythrocytic leukemia, Heilmeyer-Schner disease, acute megakaryoblastic leukemia, mast cell leukemia, panmyelosis, acute panmyeloid leukemia with myelofibrosis, lymphosarcoma cell leukemia, stemness Cell leukemia, chronic leukemia of unspecified cell type, subacute leukemia of unspecified cell type, accelerator chronic myelogenous leukemia, acute promyelocytic leukemia, acute basophil leukemia, acute eosinophilic leukemia, acute monocytic leukemia, acute myeloblastic with maturation Leukemia, acute myeloid dendritic cell leukemia, adult T-cell leukemia/lymphoma, aggressive NK cell leukemia, B-cell chronic lymphocytic leukemia, B-cell leukemia, chronic myelogenous leukemia, chronic idiopathic myelofibrosis, Kehler disease, myelomatosis, isolated Myeloma, plasma cell leukemia, angiocentric immunoproliferative lesions, lymphoid granulomatosis, angioimmunoblastic lymphadenopathy, T-gamma lymphoproliferative disease, Waldenstrom's macroglobulinemia, alpha heavy chain disease, gamma heavy chain disease and Franklin disease. It may be any one or more selected from the group consisting of

The solid cancer is colorectal cancer, melanoma, lung cancer, liver cancer, kidney cancer, stomach cancer, pancreatic cancer, rectal cancer, breast cancer, thyroid cancer, head and neck cancer, brain tumor, glioblastoma, kidney cancer, renal cell cancer, bladder cancer, and prostate cancer, acoustic neuroma ( acoustic neuroma); adenocarcinoma; adrenal gland cancer; anal cancer; angiosarcoma (eg, lymphangiosarcoma, lymphangioendotheliosarcoma, hemangiosarcoma); appendix cancer; benign monoclonal gammopathy; biliary cancer (eg, cholangiocarcinoma); bladder cancer; breast cancer (eg, adenocarcinoma of the breast, papillary carcinoma of the breast, mammary cancer, medullary carcinoma of the breast); brain cancer (eg meningioma, glioblastoma, glioblastoma (eg astrocytoma, oligodendroglioma), medulloblastoma); bronchus cancer; carcinoid tumor; cervical cancer (eg cervical adenocarcinoma); choriocarcinoma; chordoma; craniopharyngioma; colorectal cancer (eg, colon cancer, rectal cancer, colorectal adenocarcinoma); connective tissue cancer; epithelial carcinoma; ependymoma; endotheliosarcoma (eg, Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcoma); endometrial cancer (eg, uterine cancer, uterine sarcoma); esophageal cancer (eg, adenocarcinoma of the esophagus, Barrett's adenocarcinoma); Ewing's sarcoma; ocular cancer (eg intraocular melanoma, retinoblastoma); familial hypereosinophilia; gall bladder cancer; gastric cancer (eg, stomach adenocarcinoma); gastrointestinal stromal tumor (GIST); germ cell cancer; head and neck cancer (eg head and neck squamous cell carcinoma), oral cancer (eg oral squamous cell carcinoma); throat cancer ( throat cancer) (eg, laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer); heavy chain disease (eg, alpha chain disease, gamma chain disease, mu chain disease); hemangioblastoma; hypopharynx cancer; inflammatory myofibroblastic tumors; immunocytic amyloidosis; kidney cancer (eg nephroblastoma, also known as Wilms' tumor, renal cell carcinoma); liver cancer (eg, hepatocellular cancer (HCC), malignant hepatoma); lung cancer (eg, bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma of the lung); leiomyosarcoma (LMS); mastocytosis (eg, systemic mastocytosis); muscle cancer; myelodysplastic syndrome (MDS); mesothelioma; Myeloproliferative disorder (MPD) (eg, polycythemia Vera (PV), essential thrombocytosis (ET), myelofibrosis (MF), also known as unknown etiology Myeloid metaplasia (AMM), chronic idiopathic myelofibrosis, chronic myelocytic leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic syndrome, HES)); neuroblastoma; neurofibroma (eg, neurofibromatosis (NF) type 1 or type 2, schwannomatosis); neuroendocrine cancer (eg, gastroenteropancreatic neuroendocrine tumor (GEP-NET), carcinoid tumor); osteosarcoma (eg bone cancer); ovarian cancer (eg, cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma); papillary adenocarcinoma; pancreatic cancer (eg, pancreatic andenocarcinoma, intraductal papillary mucinous neoplasm (IPMN), islet cell tumors); penile cancer (eg, Paget's disease of the penis and scrotum); pinealoma; primitive neuroectodermal tumor (PNT); plasma cell neoplasia; paraneoplastic syndrome; intraepithelial neoplasm; prostate cancer (eg prostate adenocarcinoma); rectal cancer; rhabdomyosarcoma; salivary gland cancer; skin cancer (eg, squamous cell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cell carcinoma (BCC)); small bowel cancer (eg, appendix cancer); Soft tissue sarcoma (e.g., malignant fibrous histiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma); sebaceous gland carcinoma; small intestine cancer; sweat gland carcinoma, synovioma, testicular cancer (eg seminoma, testicular embryonal carcinoma); thyroid cancer (eg, papillary carcinoma of the thyroid, papillary thyroid carcinoma (PTC), medullary thyroid cancer); urethral cancer; vaginal cancer; and vulvar cancer (eg, Paget's disease of the vulva).

In one embodiment, the breast cancer may be triple negative breast cancer (TNBC).

As used herein, the term "prevention" may refer to any activity that suppresses or delays the onset of cancer in a subject by administration of a pharmaceutical composition according to one aspect.

As used herein, the term "treatment" may refer to any activity that improves or beneficially changes symptoms of cancer of a subject by administration of a pharmaceutical composition according to one aspect.

The pharmaceutical composition may be provided as a pharmaceutical composition including the active ingredient alone or including one or more pharmaceutically acceptable carriers, excipients or diluents.

Specifically, the carrier may be, for example, a colloidal suspension, powder, saline solution, lipid, liposome, microspheres or nano-spherical particles. They may be complexed with or associated with the delivery vehicle and are known in the art such as lipids, liposomes, microparticles, gold, nanoparticles, polymers, condensation reagents, polysaccharides, polyamino acids, dendrimers, saponins, adsorption enhancing substances or fatty acids. It can be delivered in vivo using known delivery systems.

When the pharmaceutical composition is formulated, it is prepared using diluents or excipients such as commonly used lubricants, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, fillers, extenders, binders, wetting agents, disintegrants, surfactants, etc. can Solid preparations for oral administration may include tablets, pills, powders, granules, capsules, etc., and such solid preparations may contain at least one excipient in the composition, for example, starch, calcium carbonate, sucrose ) or by mixing lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral use include suspensions, solutions for oral use, emulsions, syrups, etc., and various excipients such as wetting agents, sweeteners, aromatics, preservatives, etc. may be included in addition to water and liquid paraffin, which are commonly used simple diluents. . Formulations for parenteral administration may include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending agents. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycero-geratin, etc. may be used, and when prepared in the form of eye drops, known diluents or excipients may be used. there is.

Also, in one aspect, the pharmaceutical composition may be administered alone or in combination with other anticancer agents.

The pharmaceutical composition may be provided in a mixture with a conventionally known composition for preventing or treating cancer or another existing anticancer agent, and the other anticancer agent is a conventionally known composition for preventing or treating cancer, an existing anticancer agent, or a new anticancer agent. It may be an anti-cancer drug being developed.

When the pharmaceutical composition includes another anticancer agent having an effect of preventing or treating cancer, it is important to mix the amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.

As used herein, the term "combination therapy" or "combined treatment" or "in combination" refers to any form of simultaneous or concurrent treatment using at least two separate therapeutic agents.

The components of the combination therapy may be administered simultaneously, sequentially or in any order. The ingredients can be administered in a suitable manner, in different doses or at different frequencies of administration or via different routes.

The term "administered simultaneously" as used herein is not particularly limited and means that the components of the combination therapy are administered substantially simultaneously, eg as a mixture or in an immediately succeeding sequence.

The term "administered sequentially" as used herein is not particularly limited, and means that the components of the combination therapy are not administered simultaneously, but are administered one by one or in groups with a specific time interval between administrations. The time interval may be the same or different between administration of each of the components of the combination therapy, and may be selected, for example, from the range of 2 minutes to 96 hours, 1 day to 7 days or 1 week, 2 weeks or 3 weeks. . Generally, the time interval between administrations may range from several minutes to several hours, for example from 2 minutes to 72 hours, from 30 minutes to 24 hours, or from 1 to 12 hours. Additional examples include time intervals ranging from 24 to 96 hours, 12 to 36 hours, 8 to 24 hours, and 6 to 12 hours.

As used herein, the term "administration" means introducing a predetermined substance into a subject by an appropriate method, and the term "subject" refers to all organisms such as rats, mice, livestock, and the like, including humans capable of having cancer. As a specific example, it may be mammals including humans.

The route of administration of the pharmaceutical composition is, but is not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or work is included

The composition may be administered orally or parenterally, and in the case of parenteral administration, external skin or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection injection method may be selected.

The pharmaceutical composition is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level depends on the type and severity of the patient's disease, the activity of the drug, and the drug. sensitivity, time of administration, route of administration and excretion rate, duration of treatment, factors including concomitantly used drugs, and other factors well known in the medical field.

The compound of Formula 1 or a pharmaceutically acceptable salt thereof is 1 to 400 mg/kg/day, specifically 1 to 400 mg/kg/day, 1 to 300 mg/kg/day, 1 to 250 mg/kg /day, 100 to 400 mg/kg/day, 100 to 300 mg/kg/day, 100 to 250 mg/kg/day, 150 to 400 mg/kg/day, 150 to 300 mg/kg/day or 150 to 300 mg/kg/day It can be administered at 250 mg/kg/day.

Administration of the pharmaceutical composition may be administered once a day or divided into several times. Specifically, it may be administered with a 1-day break after 6-day administration on 7 days, a 2-day break after 5-day administration, or a 3-day rest cycle after 4 days administration, and more specifically, a 5-day administration followed by a 2-day break cycle. there is.

Specifically, the effective amount of the pharmaceutical composition may vary depending on the patient's age, sex, condition, body weight, absorption rate, inactivation rate, excretion rate, disease type, concomitant drug of the active ingredient in the body, administration route, obesity It may increase or decrease depending on the severity of the condition, gender, weight, age, etc.

Another aspect provides a pharmaceutical composition for inhibiting cancer metastasis comprising a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 1]

Cancer is a product of abnormal, uncontrolled and disorderly cell proliferation. In particular, it is classified as malignant if it has destructive growth, invasiveness and metastasis. The main cause of death in cancer patients is not the initial tumor, but the metastasis of cancer cells to other tissues. Looking at the metastasis process of cancer cells, first, metastatic cancer cells break away from the primary tumor, invade into support structures such as the stroma and basement membrane of normal tissue, attach to the capillary walls of other target tissues, degrade the extracellular matrix and basement membrane, and then degrade the extracellular matrix and basement membrane. It leaks into blood vessels and is accompanied by angiogenesis in new tissues, resulting in the formation of secondary tumors. Therefore, the metastasis process is composed of major steps such as migration, adhesion, and invasion, and cancer metastasis can be inhibited if any one of these steps can be prevented.

The compound represented by Formula 1 can effectively suppress cancer metastasis by inhibiting migration or invasion of cancer cells.

Another aspect provides a health functional food for preventing or improving cancer comprising a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 1]

.

.

As used herein, the term "acceptable in food science" means exhibiting characteristics that are not toxic to cells or humans exposed to the compound.

In the present specification, the term "acceptable food salt" means a salt prepared using a specific compound according to one aspect and a relatively non-toxic acid or base, and may be within the above-described range for "salt".

As used herein, the term "improvement" may refer to any activity that at least reduces a parameter related to the condition being treated, eg, the severity of a symptom. In this case, the health functional food may be used before or after the onset of the disease, simultaneously with or separately from a drug for treatment, for the prevention or improvement of cancer.

In the health functional food, the active ingredient may be added to food as it is or used together with other food or food ingredients, and may be appropriately used according to conventional methods. The mixing amount of the active ingredient can be suitably determined depending on the purpose of its use (for prevention or improvement). In general, when preparing food or beverage, the health functional food may be added in an amount of about 15% by weight or less, more specifically about 10% by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be less than the above range.

The health functional food may be formulated as one selected from the group consisting of tablets, pills, powders, granules, powders, capsules and liquid formulations by further including one or more of carriers, diluents, excipients and additives. Examples of foods to which a compound according to one aspect may be added include various foods, powders, granules, tablets, capsules, syrups, beverages, gum, tea, vitamin complexes, health functional foods, and the like.

Specific examples of the carrier, excipient, diluent, and additive include lactose, dextrose, sucrose, sorbitol, mannitol, erythritol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium phosphate, calcium silicate, and microcrystalline cellulose. , polyvinylpyrrolidone, cellulose, polyvinylpyrrolidone, methylcellulose, water, sugar syrup, methylcellulose, methyl hydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate and mineral oil It may be at least one selected from.

In addition to containing the active ingredient, the health functional food may contain other ingredients as essential ingredients without particular limitation. For example, it may contain various flavoring agents or natural carbohydrates as additional ingredients like a normal beverage. Examples of the aforementioned natural carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (thaumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can advantageously be used. can The ratio of the natural carbohydrates can be appropriately determined by a person skilled in the art.

In addition to the above, the health functional food according to one aspect is various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and salts thereof , alginic acid and its salts, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, and the like. These components can be used independently or in combination, and the ratio of these additives can also be appropriately selected by those skilled in the art.

Another aspect provides a pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 2 below or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 2]

The compound represented by Formula 1 may be named 1-(4-methoxybenzyl)-4-((2-methyl-1H-imidazol-1-yl)methyl)piperidin-4-ol, and in the following examples # 198 compounds.

Another aspect provides a pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 3 or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 3]

The compound represented by Formula 3 may be named 4-(aminomethyl)-1-(6-chloroquinolin-4-yl)piperidin-4-ol and is referred to as compound #21 in the following examples.

Another aspect provides a pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 4 or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 4]

The compound represented by Formula 4 will be named 1-(2-{ethyl[2-hydroxy-2-(3-hydroxyphenyl)ethyl]amino}ethyl)-2,3-dihydro-1H-imidazol-2-one and is referred to as #124 in the examples below.

Another aspect provides a pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 5 or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 5]

The compound represented by Formula 5 may be named 1,8-Anthracenedimethanamine and is referred to as #129 in the following Examples.

Another aspect provides a pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 6 or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 6]

The compound represented by Formula 6 will be named 3-{2-[trans-3-hydroxy-4-(hydroxymethyl)-1-piperidinyl]ethyl}-6-methyl-2,4(1H,3H)-pyrimidinedione. and is referred to as #85 in the examples below.

Another aspect provides a pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 7 or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 7]

The compound represented by Formula 7 may be named 2-amino-9-(5-(aminomethyl)-3,4-dihydroxytetrahydrofuran-2-yl)-1H-purin-6(9H)-one, and the following examples referred to as #141 in

Another aspect provides a pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 8 or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 8]

The compound represented by Formula 8 may be named 3-(aminomethyl)-N-{2-[2-(dimethylamino)acetamido]-5-methylphenyl}benzamide and is referred to as #101 in the following examples.

Another aspect provides a pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 9 or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 9]

The compound represented by Formula 9 is 2-({[(3R,4R)-4-(hydroxymethyl)-1-[4-(hydroxymethyl)-1,3-thiazol-2-yl]pyrrolidin-3-yl] methyl}(methyl)amino)ethan-1-ol and is referred to as #196 in the examples below.

Another aspect provides a pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 10 or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 10]

The compound represented by Formula 10 may be named N-{1-amino-3-[(2-methylpiperidin-2-yl)formamido]propan-2-yl}-2-methylpiperidine-2-carboxamide, and the following In the examples it is referred to as #29.

Another aspect provides a pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 11 or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 11]

The compound represented by Formula 11 may be named N-{1-amino-3-[(2-methylpiperidin-2-yl)formamido]propan-2-yl}-2-methylpiperidine-2-carboxamide, and the following In the examples it is referred to as #97.

Another aspect provides a pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 12 or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 12]

The compound represented by Formula 12 is 1-(3-(((2-hydroxyethyl)(methyl)amino)methyl)-4-(hydroxymethyl)pyrrolidin-1-yl)-2-(2-methyl-1H-imidazol -1-yl)ethanone) and is referred to as #188 in the examples below.

Another aspect provides a pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 13 or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 13]

The compound represented by Formula 13 may be named 3-({4-[2-(2-methyl-1H-imidazol-1-yl)ethyl]piperidin-1-yl}methyl)pyrrolidin-3-ol, Referred to as #89 in the Examples below.

Another aspect provides a pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 14 or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 14]

The compound represented by Formula 14 may be named 1-[(4-hydroxyphenyl)methyl]-1,2,3,4-tetrahydroisoquinoline-6,7-diol and is referred to as #110 in the following examples.

The compounds represented by Formulas 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 and 14 inhibit Gli in the same way as the compound represented by Formula 1 to prevent or treat cancer. It may work.

The terms and methods described for the above invention are equally applied between each invention.

According to the compound according to one aspect, it not only has a cancer cell-specific killing effect by inhibiting Gli, but also has an effect of suppressing cancer metastasis by inhibiting the migration and invasion of cancer cells, making it a pharmaceutical composition for preventing or treating cancer. There are effects that can be useful.

1 is a graph showing the results of Gli Reporter assay analysis confirming the Gli inhibitory ability of compound #168.
2 is a graph showing the results of confirming that compounds #198, #168, and #21 are not cytotoxic in normal cells, MRC-5.
Figure 3 is a graph of the results confirming that compounds #198, #168, and #21 are not cytotoxic in PBMC, which are normal cells.
4 is a graph showing the results of confirming that compounds #198, #168, and #21 are not cytotoxic in normal cells, VERO.
5 is a graph showing the results of confirming that compounds #198, #168, and #21 are not cytotoxic in normal cells, WI-38.
6 is a graph showing the results confirming that compound #168 inhibits the mRNA expression of Gli1, Gli2, and Cyclin D1 in melanoma cells A375.
7 is a graph showing the results confirming that compound #168 inhibits the mRNA expression of Gli1, Gli2 and Cyclin D1 in pancreatic cancer cells, PANC-1.
8 is a graph showing the results confirming that compound #168 inhibits the mRNA expression of Gli1, Gli2 and Cyclin D1 in HT-29, which is a colorectal cancer cell.
9 is a graph showing the results confirming that compound #168 inhibits the mRNA expression of Gli1, Gli2 and Cyclin D1 in melanoma cells, SK-MEL-3.
10 is a graph showing the results confirming that compound #168 inhibits the mRNA expression of Gli1, Gli2 and Cyclin D1 in breast cancer cells MDA-MB-231.
11 is a graph showing the results confirming that the positive control GANT 61 suppresses the mRNA expression of Gli1, Gli2, and Cyclin D1 in A375 melanoma cells.
12 is a graph showing the results confirming that the positive control GANT 61 suppresses the mRNA expression of Gli1, Gli2 and Cyclin D1 in pancreatic cancer cells, PANC-1.
13 is a graph showing the results confirming that the positive control GANT 61 suppresses the mRNA expression of Gli1, Gli2 and Cyclin D1 in colorectal cancer cells, HT-29.
14 is a graph showing the results confirming that the positive control GANT 61 suppresses the mRNA expression of Gli1, Gli2 and Cyclin D1 in melanoma cells, SK-MEL-3.
15 is a graph showing the results confirming that the positive control GANT 61 suppresses the mRNA expression of Gli1, Gli2 and Cyclin D1 in breast cancer cells MDA-MB-231.
16 is a photograph confirming that the #168 compound and the positive control GANT 61 inhibit p21 and Cyclin D1 protein expression in A375 melanoma cells.
17 is a photograph confirming that compound #168 and positive control GANT 61 inhibit p21 and Cyclin D1 protein expression in breast cancer cells MDA-MB-231.
18 is a photograph confirming that compound #168 and positive control GANT 61 inhibit p21 and Cyclin D1 protein expression in HT-29, which is a colorectal cancer cell.
19 is a photograph confirming that compound #168 and the positive control group GANT 61 inhibit p21 and Cyclin D1 protein expression in pancreatic cancer cells, PANC-1.
20 is a graph showing the inhibition of cell migration when melanoma cells A375 were treated with compounds #198, #168, and #21 for 24 hours.
FIG. 21 is a graph showing the inhibition of cell migration when melanoma cells A375 were treated with compounds #198, #168, and #21 for 48 hours.
22 is a photograph confirming that cell migration was inhibited when A375, which is a melanoma cell, was treated with compound #168 for 48 hours.
23 is a graph showing the inhibition of cell migration when treated with compounds #198, #168, and #21 for 24 hours in breast cancer cells, MDA-MB-231.
24 is a graph showing the inhibition of cell migration when treated with compounds #198, #168, and #21 for 48 hours in MDA-MB-231 breast cancer cells.
25 is a photograph confirming that cell migration is inhibited when the #168 compound is treated for 48 hours in breast cancer cells, MDA-MB-231.
26 is a graph showing the inhibition of cell migration when pancreatic cancer cells, PANC-1, were treated with compounds #198, #168, and #21 for 24 hours.
27 is a graph showing the inhibition of cell migration when pancreatic cancer cells, PANC-1, were treated with compounds #198, #168, and #21 for 48 hours.
28 is a photograph confirming that cell migration was inhibited when compound #168 was treated for 24 hours in pancreatic cancer cells, PANC-1.
29 is a graph showing the inhibition of cell invasion by compounds #198, #168, and #21 in A375 melanoma cells.
30 is a photograph confirming that compound #168 inhibits cell invasion in melanoma cells, A375.
31 is a graph showing the results confirming that compounds #198, #168, and #21 inhibit cell invasion in breast cancer cells, MDA-MB-231.
32 is a photograph confirming that compound #168 inhibits cell invasion in breast cancer cells, MDA-MB-231.
33 is a graph showing the inhibition of cell invasion by compounds #198, #168, and #21 in pancreatic cancer cells, PANC-1.
34 is a photograph confirming that compound #168 inhibits cell invasion in pancreatic cancer cells, PANC-1.

Hereinafter, the present invention will be described in more detail with examples, but this is only illustrative and is not intended to limit the scope of the present invention. It is apparent to those skilled in the art that the embodiments described below may be modified within a range that does not deviate from the essential gist of the invention.

Example 1. Discovery of hit compounds through virtual screening

Among the compounds derived through virtual screening, compounds with the potential to inhibit Gli proteins were selected.

Specifically, the 3D structure was used in the ZINC19 database, and a library of about 4.8 million ligands was constructed in consideration of the characteristics of the binding site by first analyzing the structure of the Gli protein. Among them, 81 compounds in the database were ordered and implemented. TOP 14 were selected by confirming the Gli1 inhibitory ability test through the Gli reporter assay in Example 2. After that, TOP 14 was treated by concentration and Gli reporter assay was performed to select TOP 3.

The 14 hit compounds selected above are respectively #129, #21, #85, #141, #101, #196, #29, #97, #89, #124, #110, #168, #198 and It is marked #188.

Example 2. Confirmation of Gli inhibitory ability through Gli Reporter assay

In order to confirm whether the selected 14 hit compounds have Gli inhibitory ability, a Gli Reporter assay was performed.

Specifically, the Gli Repoter-NIH3T3 cell line was purchased from BPS Bioscience (CA, USA) and used. NIH3T3-Gli cells with Gli transcriptional activity were dispensed in a 96 well plate at a cell concentration of 2.5Х10 ⁴ cell/mL, and after 18 hours, the cells were treated with each of the 14 hit compounds at concentrations ranging from 0 to 500 nM. After 1 hour, Sonic hedgehog peptide was treated and incubated for 24 hours. Next, Gli1 Reporter activity (%) was measured through luminescence using a Reporter assay kit (Promega, WI, USA). At this time, GANT 61, a Gli inhibitor, was used as a positive control.

IC ₅₀ values are shown in Table 1 below, and compounds #198, #168, and #21 were found to inhibit Gli the most.

1 is a graph showing the results of Gli Reporter assay analysis confirming the Gli inhibitory ability of compound #168.

Rank compound number IC ₅₀ (nM) One 198 8.562 2 168 16.0 3 21 27.8 4 124 122.2 5 85 211.7 6 188 229.7 7 29 350.2 8 89 392.9 9 141 728.9 10 129 894.8 11 196 1251 12 110 3321 13 101 26417 14 97 41738 positive control GANT61 60231

Example 3. Confirmation of cytotoxicity

Experiments were performed to determine whether the selected hit compounds were cytotoxic to normal cells and cancer cells as follows.

Specifically, as normal cells, fibroblasts MRC-5, VERO, WI-38 and peripheral blood mononuclear cells PBMC were used, and as cancer cells, colorectal cancer cells HT29, melanoma cells SK-MEL-3 and A375, pancreatic cancer cells PANC-1, AsPC-1, MIA-PaCa and BxPC-3, and breast cancer cells MDA-MB-231 were used. GANT 61, a Gli inhibitor, was used as a positive control.

Normal cells were dispensed at 5 to 6,000/well in a 96-well plate for cell culture and stabilized for 24 hours. Then, GANT 61 and each of compounds #21, #168, and #198 among the 14 hit compounds were treated at concentrations ranging from 1 to 50 uM, and incubated for 24 hours. After 24 hours, 100 μL of WST-1 (water soluble tetrazolium salt) was diluted 1/10 in media, and incubated again at 37°C for 2 hours. After incubation, absorbance was measured at 450 nm using an ELISA reader.

Cancer cells were dispensed at 5 to 6,000/well in a 96-well plate for cell culture and stabilized for 24 hours. Then, GANT 61 and each of the 14 hit compounds #21, #85, #97, #101, #110, #124, #129, #141, #168, and #198 were treated by concentration from 2 to 50 uM. and incubated for 72 hours. After 72 hours, WST-1 (water soluble tetrazolium salt) was diluted 1/10 in media, dispensed in 100 μL each, and incubated again at 37°C for 2 hours. After incubation, absorbance was measured at 450 nm using an ELISA reader.

The results are shown in Figures 2 to 5 and Tables 2 to 12. While the hit compounds were not toxic to normal cells, GANT 61 showed cytotoxicity to MRC-5 and WI-38 cells, It was confirmed that the hit compound was cytotoxic to cancer cells. Among them, it was confirmed that #198, #168, and #21 had low IC ₅₀ values and excellent anticancer effects against cancer cells.

Figure 2 is a graph showing the results confirming that compounds #198, #168, and #21 are not cytotoxic to normal cells, MRC-5; Figure 3 is a graph of the results confirming that compounds #198, #168, and #21 are not cytotoxic in PBMC, which are normal cells; Figure 4 is a graph showing the results of confirming that compounds #198, #168 and #21 are not cytotoxic in normal cells, VERO; This graph is the result of confirming that there is no toxicity.

compound cell IC ₅₀ (uM) #21 HT-29 19.44 SK-MEL-3 9.785 PANC-1 19.2 MDA-MB-231 15.09 A375 8.34 AsPC-1 30.46 BxPC-3 18.38 MIA-PaCa 5.726

compound cell IC ₅₀ (uM) #85 SK-MEL-3 341.3

compound cell IC ₅₀ (uM) #97 PANC-1 845.5

compound cell IC ₅₀ (uM) #101 HT-29 102.2 SK-MEL-3 29.64 MDA-MB-231 294

compound cell IC ₅₀ (uM) #110 SK-MEL-3 102.2 MDA-MB-231 29.64 A375 294 MIA-PaCa 27.34

compound cell IC ₅₀ (uM) #124 SK-MEL-3 640.9

compound cell IC ₅₀ (uM) #129 HT-29 96.76 SK-MEL-3 8.705 MDA-MB-231 8.194 A375 7.044 AsPC-1 8.833 BxPC-3 4.068 MIA-PaCa 2.564

compound cell IC ₅₀ (uM) #141 HT-29 134.9 BxPC-3 340.2

compound cell IC ₅₀ (uM) #168 HT-29 148.7 SK-MEL-3 14.61 PANC-1 34.85 MDA-MB-231 15.07 A375 97.14 MIA-PaCa 362.9

compound cell IC ₅₀ (uM) #198 HT-29 15.76 SK-MEL-3 21.08 PANC-1 47.56 MDA-MB-231 9.66 A375 188.2 MIA-PaCa 155.7

compound cell IC ₅₀ (uM) GANT61 HT-29 10.29 SK-MEL-3 0.9633 PANC-1 15.72 MDA-MB-231 3.731 A375 6.992 AsPC-1 16.94 BxPC-3 7.418 MIA-PaCa 5.173

Example 4. Confirmation of inhibition of target gene expression

In order to confirm whether the compound #168 inhibits the expression of target genes Gli1, Gli2, and Cyclin D1, a subgene of Gli, the following experiment was performed.

Specifically, as cancer cells, colorectal cancer cells HT29, melanoma cells SK-MEL-3 and A375, pancreatic cancer cells PANC-1, and breast cancer cells MDA-MB-231 were used. GANT 61, a Gli inhibitor, was used as a positive control. Each of the cells was divided into 7Х10 ⁵ cells in a 60 mm ³ dish for cell culture and stabilized for 24 hours. Then, each concentration of GANT 61 and the #168 compound was treated at 0, 2, and 20 uM, and incubated for 48 hours. The mRNA expression level was measured 48 hours after compound treatment.

As a result, as shown in FIGS. 6 to 10 , it was confirmed that the compound #168 suppressed the mRNA expression of Gli1, Gli2 and Cyclin D1 in cancer cells.

6 is a graph showing the results of confirming that compound #168 inhibits the mRNA expression of Gli1, Gli2 and Cyclin D1 in melanoma cells A375; 7 is a graph showing the results of confirming that compound #168 inhibits the mRNA expression of Gli1, Gli2 and Cyclin D1 in pancreatic cancer cells, PANC-1; 8 is a graph showing the results of confirming that compound #168 inhibits the mRNA expression of Gli1, Gli2 and Cyclin D1 in HT-29, which is a colorectal cancer cell; 9 is a graph showing the results of confirming that compound #168 inhibits the mRNA expression of Gli1, Gli2, and Cyclin D1 in SK-MEL-3 melanoma cells; Figure 10 is a graph showing the results confirming that compound #168 inhibits the mRNA expression of Gli1, Gli2 and Cyclin D1 in breast cancer cells MDA-MB-231; 11 is a graph showing the results confirming that the positive control GANT 61 suppresses the mRNA expression of Gli1, Gli2 and Cyclin D1 in A375 melanoma cells; 12 is a graph showing the results confirming that the positive control group GANT 61 suppresses the mRNA expression of Gli1, Gli2 and Cyclin D1 in pancreatic cancer cells, PANC-1; 13 is a graph showing the results confirming that the positive control GANT 61 suppresses the mRNA expression of Gli1, Gli2 and Cyclin D1 in colorectal cancer cells, HT-29; Figure 14 is a graph confirming that the positive control GANT 61 suppresses the mRNA expression of Gli1, Gli2 and Cyclin D1 in melanoma cells SK-MEL-3; and Figure 15 is a positive control in breast cancer cells MDA-MB-231 The graph shows the result of confirming that the control GANT 61 suppresses the mRNA expression of Gli1, Gli2 and Cyclin D1.

Example 5. Confirmation of inhibition of target protein expression

In order to confirm that the #168 compound inhibits Gli, thereby increasing the expression of p21, a lower gene, and suppressing the expression of Cyclin D1, the following experiment was performed.

Specifically, HT29, a colorectal cancer cell, A375, a melanoma cell, PANC-1, a pancreatic cancer cell, and MDA-MB-231, a breast cancer cell were used as cancer cells. GANT 61, a Gli inhibitor, was used as a positive control. Each of the cells was divided into 7Х10 ⁵ cells in a 60 mm ³ dish for cell culture and stabilized for 24 hours. Then, each concentration of GANT 61 and the #168 compound was treated at 0, 2, and 20 uM, and incubated for 72 hours. After 72 hours, each cell was harvested, proteins were extracted through lysis, separated through SDS-PAGE electrophoresis, transferred to an NC membrane, and blocked with 5% skim-milk. Subsequently, Gli1 (Cell signaling, MA, USA), Gli2 (Cell signaling, MA, USA), Cyclin D1 (Cell signaling, MA, USA), p21 (santa-cruz, CA, USA) and β-actin (sigma-aldrich , CA, USA) was attached overnight at 4°C, washed with TBST the next day, and goat anti-mouse IgG-HRP (Santa Cruz) antibody was attached for 1 hour at room temperature. The level of protein expression was confirmed using ECL (Bionote, Hwaseong, Korea) detection reagent.

As a result, as shown in FIGS. 16 to 19, it was confirmed that compound #168 increased the expression of p21 and suppressed the expression of Cyclin D1.

16 is a photograph confirming that compound #168 and positive control GANT 61 inhibit p21 and Cyclin D1 protein expression in A375 melanoma cells; 17 is a photograph confirming that compound #168 and positive control GANT 61 inhibit p21 and Cyclin D1 protein expression in breast cancer cells MDA-MB-231; Figure 18 is a photograph of the results confirming that #168 compound and positive control GANT 61 inhibit the protein expression of p21 and Cyclin D1 in colon cancer cells HT-29; and Figure 19 is #168 compound in pancreatic cancer cells PANC-1 And it is a photograph as a result of confirming that the positive control group GANT 61 suppresses the protein expression of p21 and Cyclin D1.

Example 6. Confirmation of inhibition of cell motility

In order to confirm whether the compounds #21, #168 and #198 inhibit cancer metastasis, cell migration ability was confirmed as follows.

Specifically, as cancer cells, melanoma cells A375, pancreatic cancer cells PANC-1, and breast cancer cells MDA-MB-231 were used. GANT 61, a Gli inhibitor, was used as a positive control. 70 μl of each cell line was dispensed into 12 wells with 3 x 10 ⁵ /mL ibidi culture-insert and then stabilized for 24 hours. After 24 hours, GANT 61, #21, #168, and #198 compounds were treated at 0.1, 1, and 10 uM concentrations, respectively. After 24 hours and 48 hours, respectively, cell motility was checked using a microscope.

As a result, as shown in FIGS. 20 to 28 , it was confirmed that compounds #21, #168, and #198 inhibit cell migration better than the positive control group treated with GANT 61. These results indicate that compounds #21, #168 and #198 can effectively inhibit cancer metastasis.

20 is a graph showing the inhibition of cell migration in A375 melanoma cells when treated with compounds #198, #168, and #21 for 24 hours; 21 is a graph showing the inhibition of cell migration in melanoma cells, A375, when compounds #198, #168, and #21 were treated for 48 hours; 22 is a photograph confirming that cell migration was inhibited when A375, which is a melanoma cell, was treated with compound #198 for 48 hours; 23 is a graph showing the inhibition of cell migration when treated with compounds #198, #168, and #21 for 24 hours in MDA-MB-231 breast cancer cells; 24 is a graph showing the inhibition of cell migration in breast cancer cells, MDA-MB-231, when compounds #198, #168, and #21 were treated for 48 hours; 25 is a photograph confirming that cell migration is inhibited when the #198 compound is treated for 48 hours in breast cancer cells, MDA-MB-231; 26 is a graph showing the inhibition of cell migration when pancreatic cancer cells, PANC-1, were treated with compounds #198, #168, and #21 for 24 hours; 27 is a graph showing the inhibition of cell migration when #198, #168, and #21 compounds were treated for 48 hours in pancreatic cancer cells PANC-1; and FIG. This is a photograph as a result of confirming that cell migration was inhibited when compound #198 was treated for 24 hours.

Example 7. Confirmation of inhibition of cell invasion

In order to confirm whether the compounds #21, #168 and #198 inhibit invasiveness of cancer cells, the following experiment was performed.

Specifically, after completely dissolving Matrigel at 4° C. for 16 to 20 hours, Matrigel was diluted with cold serum-free media (4° C.) at a ratio of 1:3. 50 μL of the Matrigel mixture was put into an 8.0 mm 24-well transwell insert and solidified in a 37° C. incubator for 30 minutes. 800 μL of serum-free media was added to the down chamber, and 3X10 ⁵ cells/well, 3X10 5 cells/well, 2X10 ⁵ cells/well, respectively, melanoma cell A375, pancreatic cancer cell PANC-1, and breast cancer cell MDA-MB- ²³¹ into the upper chamber. Cells/well were divided according to the number of cells. Then, an untreated control group (CN), which was not treated with anything, was set aside, and the positive control GANT 61 and the compounds #21, #168, and #198 were treated by concentration at 0.1, 1, and 10 uM, respectively, for 72 hours, 60 hours, incubated for 24 hours. Then, after removing non-infiltrated cells, cell invasion was confirmed using Diff Qick kit.

Then, after removing non-infiltrated cells, the cells were stained with 0.5% crystal-violet solution for 5 minutes, and the number of stained cells was measured.

As a result, as shown in FIGS. 29 to 34 , it was confirmed that compounds #21, #168, and #198 inhibited cancer cell invasion more significantly than the GANT 61-treated positive control group. These results indicate that compounds #21, #168 and #198 can effectively inhibit invasion of cancer cells.

29 is a graph showing the inhibition of cell invasion by compounds #198, #168, and #21 in A375 melanoma cells; 30 is a photograph confirming that compound #168 inhibits cell invasion in A375 melanoma cells; Figure 31 is a graph showing the results confirming that compounds #198, #168, and #21 inhibit cell invasion in breast cancer cells, MDA-MB-231; 32 is a photograph confirming that compound #168 inhibits cell invasion in breast cancer cells, MDA-MB-231; Figure 33 is a graph confirming that compounds #198, #168, and #21 inhibit cell invasion in pancreatic cancer cells, PANC-1; and Figure 34 is a graph showing compound #168 inhibits cell invasion in pancreatic cancer cells, PANC-1. This is a picture of the result of confirming what was ordered.

Claims (10)

A pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient:
[Formula 1]

. The pharmaceutical composition according to claim 1, which inhibits Gli (glioma-associated oncogene homolog) gene or Gli protein activity. The pharmaceutical composition according to claim 2, wherein the Gli is Gli1 or Gli2. The pharmaceutical composition according to claim 1, wherein the cancer is a cancer in which Gli is expressed. The pharmaceutical composition according to claim 1, wherein the cancer is blood cancer or solid cancer. The pharmaceutical composition according to claim 5, wherein the blood cancer is any one or more selected from the group consisting of leukemia, lymphoma, and myeloma. The method according to claim 5, wherein the solid cancer is colorectal cancer, melanoma, lung cancer, liver cancer, kidney cancer, stomach cancer, pancreatic cancer, rectal cancer, breast cancer, thyroid cancer, head and neck cancer, brain tumor, glioblastoma, kidney cancer, renal cell cancer, bladder cancer And at least one selected from the group consisting of prostate cancer, a pharmaceutical composition. The pharmaceutical composition according to claim 7, wherein the breast cancer is triple negative breast cancer (TNBC). A pharmaceutical composition for inhibiting cancer metastasis comprising a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient:
[Formula 1]

. A health functional food for preventing or improving cancer comprising a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient:
[Formula 1]

.

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