DE112018004197T5 - COMPOSITION FOR PREVENTING OR TREATING CANCER, INCLUDING TRIAZOLPYRIDINE-BASED DERIVATIVE AS AN ACTIVE SUBSTANCE - Google Patents

Abstract

The present invention relates to a composition for the prevention or treatment of cancer, comprising a triazolpyridine-based derivative or a pharmaceutically acceptable salt thereof, as an active ingredient, wherein triazolpyridine-based derivative inhibits the activity of the tankyrase enzyme to reduce the expression of the tumor factor β- Suppressing catenin, which increases cancer cell death by apoptosis and, when used in combination with anti-cancer agents or radiation, excellently lowers cell viability, and thus triazolopyridine-based derivative can be useful as a pharmaceutical composition for the prevention and treatment of cancer, a composition for enhancement the effects of radiation therapy or as a healthy functional food to prevent and improve cancer.

Description

Technical field

The present invention relates to a pharmaceutical composition for preventing or treating cancer, a composition for increasing an anti-cancer activity, or a healthy food for preventing or improving cancer, comprising a triazolpyridine-based derivative as an active ingredient.

State of the art

Although many diseases are treated and prevented through the development of modern medicine, cancer is still one of the most difficult diseases to treat. Cancer is currently the leading cause of death and is constantly increasing.

Cancer is treated through chemotherapy, surgical therapy, and radiation therapy. Of these, chemotherapy using an anti-cancer agent is most commonly used for the treatment of cancer. Around 60 different anti-cancer drugs are currently used in clinical practice, and as facts regarding cancer development and the properties of cancer cells are well known, new anti-cancer drugs are constantly being developed. However, most of the anti-cancer drugs currently used in clinical practice are often accompanied by side effects such as nausea, vomiting, ulcers in the oral cavity and small intestine, diarrhea, hair loss and bone marrow suppression, with the production of blood-active ingredients being reduced. For example, mitomycin-C is known to have a side effect such as kidney failure, and adriamycin is known to have a side effect such as myelosuppression. In particular, cisplatin, the most useful drug among the anti-cancer drugs developed to date, is generally used for the treatment of testicular cancer, ovarian cancer, lung cancer, head and neck cancer, bladder cancer, stomach cancer and cervical cancer, but has been found to have side effects including hematopoietic toxicity such as anemia , gastrointestinal toxicity such as vomiting, nausea, kidney toxicity such as renal tubular damage to the kidney, hearing loss, electrolyte abnormalities in the body, shock, peripheral nerve abnormalities have become a major problem, the development of new anti-cancer drugs with excellent safety is urgently needed.

In addition, as the number of cancer patients treated with radiation therapy increases each year, the importance of radiation therapy in cancer treatment is also increasing.

However, it was highlighted that a problem of lowering the effectiveness of radiation therapy arises due to the resistance of cancer cells to radiation and damage to normal tissues during high dose radiation treatments. Therefore, radiation therapy sensitizers have been researched to improve the effectiveness of radiation therapy.

Radiation therapy sensitizers that have been reported to date are predominantly anti-cancer drugs, for example taxol and cisplatin have been reported to be used as radiation therapy sensitizers in solid cancers such as breast cancer, uterine cancer, lung cancer, gastric cancer and colon cancer.

In addition, there is a tirapazamine as an enhancer of radiation therapy, which has no anti-cancer properties and is only used for radiation therapy, but this is only effective in tumor cells from hypoxia and inadequate drug delivery in tumor tissue due to the intra-tumor pressure specific for hypoxia was in clinical radiation therapy known to be ineffective, and these radiation therapy sensitizers have high side effects and thus a problem of their limited use.

Although there is a difference in sensitivity to radiation treatment for cancer cells if the sensitivity can be improved, the same therapeutic effect can be achieved by irradiating the patient with a lower dose of radiation, and therefore there is a need to develop anticancer drugs that treat cancer and can increase the sensitivity of cancer cells to radiation.

epiphany

Technical problem

The object of the present invention is to solve the side effects of the conventional anti-cancer drugs or radiation therapy and to provide a composition for improving the therapeutic anti-cancer activity which can improve the therapeutic anti-cancer activity.

Technical solution

in chemischer Formel 1
kann jedes von R¹ und R² dasselbe oder unterschiedlich und ausgewählt aus Wasserstoff, CN, NO₂, CF₃, Halogen und Acetyl sein und X ist beliebig ausgewählt aus Stickstoff und Kohlenstoff.The present invention provides a triazole pyridine based derivative represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof:

in chemical formula 1
each of R ¹ and R ^{2 may be the} same or different and selected from hydrogen, CN, NO ₂ , CF ₃ , halogen and acetyl, and X is arbitrarily selected from nitrogen and carbon.

in chemischer Formel 1,
kann jedes von R¹ und R² dasselbe oder unterschiedlich und ausgewählt aus Wasserstoff, CN, NO₂, CF₃, Halogen und Acetyl sein und X ist beliebig ausgewählt aus Stickstoff und Kohlenstoff.The present invention provides a pharmaceutical composition for the prevention or treatment of cancer, comprising a triazolopyridine-based derivative represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, as an active ingredient:

in chemical formula 1,
each of R ¹ and R ^{2 may be the} same or different and selected from hydrogen, CN, NO ₂ , CF ₃ , halogen and acetyl, and X is arbitrarily selected from nitrogen and carbon.

The present invention provides a composition for enhancing an anti-cancer activity, comprising a triazole pyridine-based derivative represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, as an active ingredient.

In addition, the present invention provides a healthy food for preventing or ameliorating cancer, comprising a triazolopyridine-based derivative represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, as an active ingredient.

Beneficial effects

According to the present invention, the triazole pyridine-based derivative represented by Chemical Formula 1 inhibits the activity of the tankyrase enzyme to suppress the expression of the tumor factor β-catenin, thereby increasing cancer cell death by apoptosis. When used in combination with anti-cancer agents or radiation, cell viability deteriorates excellently, and therefore the triazolpyridine-based derivative can be useful as a pharmaceutical composition for the prevention and treatment of cancer, a composition for enhancing the action of the anti-cancer agent or radiation treatment, or for one healthy functional foods can be used to prevent and improve cancer.

Figure list

• 1 shows the results of confirmation of the concentration-dependent inhibition of TNKS activity by treatment with the triazolopyridine derivative compound 1 or connection 2nd at different concentrations in vitro.
• 2nd shows the results of confirmation of inhibition of β-catenin protein expression and elevation of tumor suppressor AXIN2 after compound treatment 1 or connection 2nd which is a triazolopyridine derivative in DLD-1 colon cancer cells.
• 3rd shows the results of confirmation of inhibition of expression of VEGF2 and Birc / survivin, which are sub-genes of β-catenin, after treatment with compound 1 or connection 2nd which is a triazolopyridine derivative in DLD-1 colon cancer cells.
• 4th shows the results of confirmation of cell viability after the combination treatment with compound 1 or connection 2nd , each of which is a triazolopyridine derivative, when irradiated on DLD-1 colon cancer cells, in which APC (adenomatous polyposis coli) gene is mutated.
• 5 shows the results of confirmation of the decrease in cell viability after the combination treatment with the anti-cancer agent 5-FU and triazolopyridine derivative compound 1 or connection 2nd , of two colon cancer cell lines in which the APC gene was mutated.
• 6 shows the results of confirmation of the reduction in CYR61 and CTGF, which are subgenes of tumor factor YAP / TAZ, after treatment with compound 1 or connection 2nd that are triazolopyridine derivative in DLD-1 colon cancer cell lines.
• 7 shows the results of a confirmation decrease in cell viability after treatment with compound 1 or connection 2nd which are a triazolopyridine derivative, at various concentrations, in the SKMEL28 melanoma cell line and SKMEL28 cell line with the resistance of vemurafenib, a BRAF-inhibitory anti-cancer agent.

Best mode

in chemischer Formel 1 kann jedes von R¹ und R² dasselbe oder unterschiedlich und ausgewählt aus Wasserstoff, CN, NO₂, CF₃, Halogen und Acetyl sein und X kann ein beliebiges, ausgewählt aus Stickstoff und Kohlenstoff sein.The present invention can provide a triazole pyridine-based derivative represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof:

in chemical formula 1, each of R ¹ and R ^{2 may be the} same or different and selected from hydrogen, CN, NO ₂ , CF ₃ , halogen and acetyl, and X may be any one selected from nitrogen and carbon.

Each of R ¹ and R ^{2 of} the compound may be the same or different and selected from the group consisting of CN, NO ₂ , CF ₃ and halogen, and X may be carbon.

The triazolpyridine-based derivative can be selected from the group consisting of N - ([1,2,4,] triazole [4,3-a] pyridin-3-yl) -1- (2-cyanophenyl) piperidin-4- carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (2-nitrophenyl) piperidine-4-carboxamide, N - ([1,2,4] Triazole [4,3-a] pyridin-3-yl) -1-phenylpiperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (4-nitrophenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (2-nitro-4- (trifluoromethyl) phenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (4- (trifluoromethyl) phenyl) piperidine-4-carboxamide, N- ( [1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (4-fluoro-2-nitrophenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (4-acetylphenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl ) -1- (4-chloro-2-nitrophenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (4-cyanophenyl ) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (5-nitropyridin-2-yl) piperidine-4-carboxamide and N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (2-cyano-4-nitrophenyl) piperidine-4-carboxamide.

in chemischer Formel 1 kann jedes von R¹ und R² dasselbe oder unterschiedlich und ausgewählt aus Wasserstoff, CN, NO₂, CF₃, Halogen und Acetyl sein und X kann ein beliebiges, ausgewählt aus Stickstoff und Kohlenstoff sein.The present invention can provide a pharmaceutical composition for preventing or treating cancer, comprising a triazole pyridine-based derivative represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, as an active ingredient:

in chemical formula 1, each of R ¹ and R ^{2 may be the} same or different and selected from hydrogen, CN, NO ₂ , CF ₃ , halogen and acetyl, and X may be any one selected from nitrogen and carbon.

In particular, each of R ¹ and R ^{2 may be the} same or different and selected from the group consisting of CN, NO ₂ , CF ₃ and halogen, and X may be carbon.

The triazolpyridine-based derivative can be selected from the group consisting of N - ([1,2,4,] triazole [4,3-a] pyridin-3-yl) -1- (2-cyanophenyl) piperidin-4- carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (2-nitrophenyl) piperidine-4-carboxamide, N - ([1,2,4] Triazole [4,3-a] pyridin-3-yl) -1-phenylpiperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (4-nitrophenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (2-nitro-4- (trifluoromethyl) phenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (4- (trifluoromethyl) phenyl) piperidine-4-carboxamide, N- ( [1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (4-fluoro-2-nitrophenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (4-acetylphenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl ) -1- (4-chloro-2-nitrophenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (4-cyanophenyl ) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (5-nitropyridin-2-yl) piperidine-4-carboxamide and N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (2-cyano-4-nitrophenyl) piperidine-4-carboxamide.

The cancer may be one selected from the group consisting of lung cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, breast cancer, stomach cancer, liver cancer, colon cancer, skin cancer, head or neck cancer, uterine cancer, ovarian cancer, Brain cancer, larynx cancer, prostate cancer, bladder cancer, esophageal cancer, thyroid cancer, kidney cancer, blood cancer and rectal cancer.

In particular, the triazole pyridine-based derivatives can be used effectively for the treatment of cancer cells in which β-catenin, a tumor factor, is overexpressed by inhibiting the activity of tankyrase (TNKS) enzymes.

The TNKS has several binding protein partners containing a double-stranded telomer repeat binding protein TRF1 (telomeric repeat binding factor 1 ); NuMA (NuclearMitotic Associated Protein), which is an essential protein in a mitotic spindle arrangement; IRAP (aminopeptidase), an intrinsic membrane protein involved in glucose uptake corresponding to insulin; and Mcl-1 (myeloid leukemia cell differentiation protein), which is an apoptosis-promoting protein and shows biological function through its various interactions.

According to one embodiment of the present invention, it is confirmed that triazole pyridine-based compounds 1 to 12, as shown in Table 1, inhibit the activity of the TNKS-1 enzyme but do not inhibit the activity of PARP-1, and thus the activity of the triazole pyridine -based derivatives specific for the TNKS-1 enzyme.

From the above results of the treatment of DLD-1 colon cancer cells in which the APC gene was mutated with the triazole pyridine-based compounds 1 and 2nd , identified as TNKS enzyme activity inhibitors, was confirmed to be as described in 2nd demonstrated expression of β-catenin protein by compound 1 and connection 2nd was decreased while the expression of AXIN2 protein was increased. Therefore, the triazolopyridine derivatives inhibited the activity of tankyrase (TNKS) enzymes, which resulted in the expression of β-catenin, and it can be provided as an anti-cancer agent for increasing cancer cell death by apoptosis.

In one embodiment of the present invention, the pharmaceutical composition for preventing or treating cancer, comprising the compound represented by Chemical Formula 1, or a pharmaceutically acceptable salt thereof, as an active ingredient, in one of the formulations selected from the group from injections, granules, powders, tablets, pills, capsules, suppositories, gels, suspensions, emulsions, drops or solutions according to a conventional method.

Another embodiment of the invention may further comprise at least one additive selected from the group consisting of carriers, auxiliaries, disintegrants, sweeteners, coating agents, swelling agents, lubricants, lubricants, flavors, antioxidants, buffering agents, bacteriostatics, diluents, dispersants, surfactants, binders and Lubricants which are conveniently used for the preparation of the pharmaceutical composition.

Examples of carriers, auxiliaries and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzene Propyl hydroxybenzoate, talc, magnesium stearate and mineral oil. Solid formulations for oral administration contain tablets, pills, powder, granules, capsules, etc., and such solid formulations may contain at least one adjuvant such as starch, calcium carbonate, sucrose or lactose, gelatin and the like in addition to the composition. In addition to simple aids, lubricants such as magnesium stearate and talc are also used. Examples of liquid formulations for oral administration include suspensions, solutions, emulsions, syrups and the like, and various auxiliaries such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are generally used as simple diluents. Formulations for parenteral administration contain sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories and the like. Examples of the non-aqueous solution and the suspension include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable esters such as ethyl oleate and the like. As the base of the suppository, Witepsol, Macrogol, Tween 61, cocoa butter, laurinum, glycerogelatin and the like can be used.

According to one embodiment of the invention, the pharmaceutical composition can be administered to the subject intravenously, intraarterially, intraperitoneally, intramuscularly, intrasternally, transdermally, nasally, inhaled, topically, rectally, orally, intraocularly or intradermally in a conventional manner.

The preferred dosage of the compound represented by Chemical Formula 1 may vary depending on the condition and weight of the subject, the type and extent of the disease, the drug form, the route of administration and the duration, and may be appropriately selected by those skilled in the art. According to an embodiment of the present invention, the daily dosage can be 0.01 to 200 mg / kg, in particular 0.1 to 200 mg / kg, more particularly 0.1 to 100 mg / kg, but is not limited to this. Administration can be administered once a day or divided into several times, and the scope of the invention is not restricted thereby.

In the present invention, the "subject" can be, but is not limited to, a mammal containing a human.

In addition, the present invention can provide a composition for enhancing an anti-cancer activity, comprising a triazole pyridine-based derivative represented by Chemical Formula 1, or a pharmaceutically acceptable salt thereof, as an active ingredient.

The anti-cancer enhancing composition can be treated in combination with an anti-cancer agent or radiation.

The anti-cancer agent may be one or more selected from the group consisting of cisplatin, 5-fluorouracil, paclitaxel, doxorubicin, daunorubicin, vinblastine, vincristine, actinomycin D, teniposide, etoposide, cyclophosphamide, epirubicin, adriamycin, daunomycin and mitomycin.

The composition may contain 1 to 99 parts by weight of the anti-cancer agent and 1 to 99 parts by weight of the triazole pyridine-based derivative represented by Chemical Formula 1 based on 100 parts by weight of the entire composition.

The anticancer enhancement composition may increase the anticancer activity against cancer selected from the group consisting of lung cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, breast cancer, gastric cancer, liver cancer, colon cancer, skin cancer, head cancer or throat cancer, uterine cancer, ovarian cancer, brain cancer, larynx cancer, prostate cancer, bladder cancer, esophageal cancer, thyroid cancer, kidney cancer, blood cancer and rectal cancer.

According to another embodiment of the present invention, the cultured DLD-1 colon cancer cells were linked 1 and connection 2nd Treated at a concentration of 20 µM and then irradiated with 3 Gy gamma radiation and cultured for 3 days to determine cell viability, and it was confirmed that the combination treatment of triazole pyridine-based compound 1 or connection 2nd and radiation reduced the survival rate of cancer cells, as in 4th shown. In addition, DLD-1 cells and SW480 cells were connected 1 or connection 2nd at a concentration of 20 µM in combination with 5-FU (10 µM), a colon cancer treatment anticancer agent, treated for 48 hours and an MTT assay was performed to confirm cell viability and as in 5 shown, it was confirmed that the effect on inhibiting cell proliferation of the experimental group, the 5-FU in combination with compound 1 and connection 2nd was increased 2-fold or 1.5-fold compared to the control treated with 5-FU as an anti-cancer agent.

From the foregoing, it is confirmed that it has been confirmed that the triazole pyridine-based compound increases the anti-cancer therapeutic effect when used in combination with an anti-cancer agent or radiation that the triazole pyridine-based compound can be used as an anti-cancer adjuvant.

In addition, the present invention can provide a healthy food for cancer prevention or improvement, comprising a triazole pyridine-based derivative represented by Chemical Formula 1, or a pharmaceutically acceptable salt thereof, as an active ingredient.

The cancer can be a cancer selected from the group consisting of lung cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, breast cancer, stomach cancer, liver cancer, colon cancer, skin cancer, head or neck cancer, uterine cancer, ovarian cancer, Brain cancer, larynx cancer, prostate cancer, bladder cancer, esophageal cancer, thyroid cancer, kidney cancer, blood cancer and rectal cancer.

The healthy food can be used with other foods or supplements in addition to the triazolopyridine derivatives represented by Chemical Formula 1, or pharmaceutically acceptable salts thereof, and can be used appropriately according to conventional methods. The mixed amount of the active ingredient can be appropriately selected depending on the purpose, for example, prophylactic, health or therapeutic treatment.

The effective dose of the compound contained in the healthy food may be used according to the effective dose of the therapeutic agent, but may be less than the above range in the case of prolonged health or hygienic or health-promoting use. It is obvious that the component can be used in an amount above the range because there is no safety problem.

There is no particular restriction on the type of healthy food, for example, meat, sausage, bread, chocolate, confectionery, snacks, confectionery, pizza, ramen, other pasta, chewing gum, dairy products containing ice cream, various soups, juices, tea, Contain drinks, alcoholic beverages, vitamin complexes, etc.

Furthermore, the present invention includes a triazole pyridine-based derivative represented by Chemical Formula 1, or a pharmaceutically acceptable salt thereof, as an active ingredient, and the triazole pyridine-based derivative or a pharmaceutically acceptable salt thereof can be used as a reagent composition for inhibiting the activity of Tankyrase enzyme can be provided, characterized by inhibiting the activity of the tankyrase (TNKS) enzyme of the cell in vitro.

In addition, the present invention can provide a method for inhibiting the activity of tankyrase enzyme in vitro, comprising the step of treating a cancer cell with triazolopyridine-based derivative represented by Chemical Formula 1, or a pharmaceutically acceptable salt thereof.

In the following, the present invention will be described in detail with reference to the following examples. The examples are only for the more detailed description of the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not limited to these exemplary embodiments according to the spirit of the present invention.

<Synthesis example 1> Synthesis of triazolopyridine derivatives

1. 1. N-([1,2,41Triazol[4,3-alpyridin-3-vl)-1-(2-cyanophenyl)piperidin-4-carboxamid (Verbindung 1)
   
   Wie im oben stehenden Reaktionsschema 1 dargestellt, wurden 21 mg von Verbindung 1 (3) mit einer Ausbeute von etwa 25% durch Amidkopplungsreaktion eines Ausgangsmaterials (1) und von Carbonsäure (2) unter Verwendung von EDCI und HOBt in Gegenwart von Trimethylamin und Dichlormethan-Lösemittel erhalten. ¹H NMR (DMSO-d6, 400 MHz): δ 10,96 (1H, s), 7,96 (1H, d, J = 8,0Hz), 7,73 (1H, m), 7,61 (1H, t, J = 8,0Hz), 7,4 (1H, ddd, J = 12,0, 4,0 und 0,8Hz), 7,22 (1H, d, J = 8,0Hz), 7,11 (1H, t, J = 8,0Hz), 6,99 (1H, t, J= 6,0Hz), 3,58 (2H, d, J=12,0Hz), 2,91 (2H, t, J = 12,0Hz), 2,50 (1H, m) 2,10 (2H, d, J = 12,0Hz), 1,91 (2H, t, J = 6,0Hz)
2. 2. N-([1,2,41Triazol[4,3-alpyridin-3-yl)-1-(2-nitrophenyl)piperidin-4-carboxamid (Verbindung 2)
   
   Wie im oben stehenden Reaktionsschema 2 gezeigt, wurden 25 mg von Verbindung 2 (3) mit einer Ausbeute von etwa 29% durch Amidkopplungsreaktion eines Ausgangsmaterials (1) und von Carbonsäure (2) unter Verwendung von EDCI und HOBt in Gegenwart von Trimethylamin und Dichlormethan-Lösemittel erhalten. ¹H NMR (DMSO-d6, 400 MHz): δ 10,95 (1H, s), 7,95 (1H, d, J = 8,0Hz), 7,81 (1H, dd, J = 8,0 und 4,0Hz), 7,75 (1H, d, J = 8,0Hz), 7,59 (1H, td, J = 8,0, und 4,0Hz), 7,38 (2H, m), 7,13 (1H, t, J = 8,0Hz), 7,00 (1H, t, J = 4,0Hz), 3,27 (2H, d, J = 12,0Hz), 2,91 (2H, t, J = 12,0Hz), 2,71 (1H, m), 2,03 (2H, d, J = 12,0Hz), 1,85 (2H, m)
3. 3. N-([1,2,41Triazol[4,3-alpyridin-3-yl)-1-phenylpiperidin-4-carboxamid (Verbindung 3)
   
   Wie im oben stehenden Reaktionsschema 3 gezeigt, wurden, 24 mg von Verbindung 3 (3) mit einer Ausbeute von etwa 28% durch Amidkopplungsreaktion eines Ausgangsmaterials (1) und von Carbonsäure (2) unter Verwendung von EDCI und HOBt in Gegenwart von Trimethylamin und Dichlormethan-Lösemittel erhalten. ¹H NMR (CDCl₃-d, 400 MHz): δ 11,65 (1H, s), 7,91 (1H, d, J = 4,0Hz), 7,67 (1H, d, J = 8,0Hz), 7,27 (3H, m), 6,97 (2H, d, J = 8,0Hz), 6,9 (2H, m), 3,76 (2H, dt, J=11,0 und 3,1 Hz), 2,9 (3H, td, J=10,0 und 3,2Hz), 2,19 (2H, dd, J=14,0 und 2,0Hz), 2,07(2H, ddd, J=24,0, 12,0 und 4,0Hz)
4. 4. N-([1,2,41Triazol[4,3-alpyridin-3-yl)-1-(4-nitrophenyl)piperidin-4-carboxamid (Verbindung 4)
   
   Wie im oben stehenden Reaktionsschema 4 gezeigt, wurden 22 mg von Verbindung 4 (3) mit einer Ausbeute von etwa 25% durch Amidkopplungsreaktion eines Ausgangsmaterials (1) und von Carbonsäure (2) unter Verwendung von EDCI und HOBt in Gegenwart von Trimethylamin und Dichlormethan-Lösemittel erhalten. ¹H NMR (DMSO-d6, 400 MHz): δ 10,96 (1H, s), 8,06 (2H, d, J = 8,0Hz), 7,93 (1H, d, J = 4,0Hz), 7,74 (1H, d, J = 8,0Hz), 7,39 (1H, ddd, J = 8,4, 6,6 und 1,8Hz), 7,07 (2H, d, J = 8,0Hz), 6,97 (1H, t, J = 6,0Hz), 4,13 (2H, d, J = 12,0Hz), 3,14 (2H, t, J = 12,0Hz), 2,89 (1H, m), 2,05 (2H, d, J = 12,0Hz), 1,75 (2H, m)
5. 5. N-([1,2,41Triazol[4,3-alpyridin-3-yl)-1-(2-nitro-4-(trifluormethyl)phenyl) piperidin-4-carboxamid (Verbindung 5)
   
   Wie im oben stehenden Reaktionsschema 5 gezeigt, wurden 28 mg von Verbindung 5 (3) mit einer Ausbeute von etwa 27% durch Amidkopplungsreaktion eines Ausgangsmaterials (1) und von Carbonsäure (2) unter Verwendung von EDCI und HOBt in Gegenwart von Trimethylamin und Dichlormethan-Lösemittel erhalten. ¹H NMR (DMSO-d6, 400 MHz): δ 10,97 (1H, s), 8,16 (1H, d, J = 4,0Hz), 7,87 (1H, d, J = 4,0Hz), 7,85 (1H, d, J = 2,0Hz), 7,75 (1H, dt, J = 10,6 und 0,4Hz), 7,49 (1H, d, J = 8,0Hz), 7,40 (1H, ddd, J = 10,2, 6,0 und 1,4Hz), 6,99 (1H, td, J = 7,5, 7,5 und 3,2Hz), 3,43 (2H, d, J = 16,0Hz), 3,07 (2H, t, J = 12,0Hz), 2,79 (1H, m), 2,05 (2H, d, J = 12,0Hz), 1,85 (2H, t, J = 12,0Hz)
6. 6. N-([1,2,41Triazol[4,3-alpyridin-3-yl)-1-(4-(trifluormethvl)phenyl) piperidin-4-carboxamid (Verbindung 6)
   
   Wie im oben stehenden Reaktionsschema 6 gezeigt, wurden 24 mg von Verbindung 6 (3) mit einer Ausbeute von etwa 27% durch Amidkopplungsreaktion eines Ausgangsmaterials (1) und von Carbonsäure (2) unter Verwendung von EDCI und HOBt in Gegenwart von Trimethylamin und Dichlormethan-Lösemittel erhalten. ¹H NMR (DMSO-d6, 400 MHz): δ 10,94 (1H, s), 7,93 (1H, d, J = 8,0Hz), 7,74 (1H, d, J = 12,0Hz), 7,50 (2H, d, J = 12,0Hz), 7,39 (1H, t, J = 8,0Hz), 7,10 (2H, d, J = 8,0Hz), 6,97 (1H, t, J = 6,0Hz), 3,97 (2H, d, J = 16,0Hz), 2,95 (2H, t, J = 12,0Hz), 2,81 (1H, m), 2,02 (2H, d, J = 12,0Hz), 1,76 (2H, m)
7. 7. N-([1,2,41Triazol[4,3-alpyridin-3-y1)-1-(4-fluor-2-nitrophenyl)piperidin-4-carboxamid (Verbindung 7)
   
   Wie im oben stehenden Reaktionsschema 7 gezeigt, wurden 28 mg von Verbindung 7 (3) mit einer Ausbeute von etwa 27% durch Amidkopplungsreaktion eines Ausgangsmaterials (1) und von Carbonsäure (2) unter Verwendung von EDCI und HOBt in Gegenwart von Trimethylamin und Dichlormethan-Lösemittel erhalten. ¹H NMR (DMSO-d6, 400 MHz): δ 10,94 (1H, s), 7,94 (1H, d, J = 4,0Hz), 7,83 (1H, dd, J = 8,0 und 4,0Hz), 7,75 (1H, d, J = 8,0Hz), 7,52 (2H, m), 7,40 (1H, dd, J = 12,0 und 6,0Hz), 6,99 (1H, t, J = 8,0Hz), 3,20 (2H, d, J = 12,0Hz), 2,88 (2H, t, J = 12,0Hz), 2,69 (1H, m), 2,03 (2H, d, J = 8,0Hz), 1,82 (2H, m)
8. 8. N-([1,2,41Triazol[4,3-alpyridin-3-yl)-1-(4-acetylphenyl)piperidin-4-carboxamid (Verbindung 8)
   
   Wie im oben stehenden Reaktionsschema 8 gezeigt, wurden 24 mg von Verbindung 8 (3) mit einer Ausbeute von etwa 28% durch Amidkopplungsreaktion eines Ausgangsmaterials (1) und von Carbonsäure (2) unter Verwendung von EDCI und HOBt in Gegenwart von Trimethylamin und Dichlormethan-Lösemittel erhalten. ¹H NMR (DMSO-d6, 400 MHz): δ 10,95 (1H, s), 7,93 (1H, d, J = 8,0Hz), 7,81 (2H, d, J = 8,0Hz), 7,74 (1H, d, J = 12,0Hz), 7,39 (1H, ddd, J = 8,4, 6,4 und 1,6Hz), 6,99 (3H, m), 4,04 (2H, d, J = 12,0Hz), 3,00 (2H, t, J = 12,0Hz), 2,84 (1H, m), 2,45 (3H, s), 2,02 (2H, d, J = 12,0Hz), 1,74 (2H, m)
9. 9. N-([1,2,4]Triazol[4,3-alpyridin-3-yl)-1-(4-chlor-2-nitrophenyl)piperidin-4-carboxamid (Verbindung 9)
   
   
   Wie im oben stehenden Reaktionsschema 9 gezeigt, wurden 28 mg von Verbindung 9 (3) mit einer Ausbeute von etwa 26% durch Amidkopplungsreaktion eines Ausgangsmaterials (1) und von Carbonsäure (2) unter Verwendung von EDCI und HOBt in Gegenwart von Trimethylamin und Dichlormethan-Lösemittel erhalten. ¹H NMR (DMSO-d6, 400 MHz): δ 10,95 (1H, s), 7,95 (2H, m), 7,75 (1H, d, J = 8,0Hz), 7,65 (1H, dd, J = 12,0 und 8,0Hz), 7,39 (2H, m), 6,99 (1H, t, J = 8,0Hz), 3,26 (2H, d, J = 12,0Hz), 2,92 (2H, t, J = 12,0Hz), 2,72 (1H, m), 2,03 (2H, d, J = 8,0Hz), 1,83 (2H, m)
10. 10. N-([1,2,4]Triazol[4,3-alpyridin-3-yl)-1-(4-cyanophenyl)piperidin-4-carboxamid (Verbindung 10)
    
    Wie im oben stehenden Reaktionsschema 10 gezeigt, wurden 20 mg von Verbindung 10 (3) mit einer Ausbeute von etwa 23% durch Amidkopplungsreaktion von Ausgangsmaterial (1) und Carbonsäure (2) unter Verwendung von EDCI und HOBt in Gegenwart von Trimethylamin und Dichlormethan-Lösemittel erhalten. ¹H NMR (DMSO-d6, 400MHz): δ 10,94(1H, s), 7,93(1H, d, J=8,0Hz), 7,74(1H, d, J=8,0Hz), 7,58(1H, d, J=8,0Hz), 7,39(1H, dd, J=8,0 und 8,0Hz), 7,06(2H, d, J=8,0Hz), 6,97(1H, m), 4,03(2H, d, J=12,0Hz), 3,01(2H, m), 2,83(1H, m), 2,02(2H, d, J=16Hz), 1,73(2H, m)
11. 11. N-([1,2,41Triazol[4,3-alpyridin-3-yl)-1-(5-nitropyridin-2-yl)piperidin-4-carboxamid (Verbindung 11)
    
    Wie im oben stehenden Reaktionsschema 11 gezeigt, wurden 20 mg von Verbindung 11 (5) mit einer Ausbeute von etwa 26% durch Amidkopplungsreaktion von Ausgangsmaterial (1) und Carbonsäure (2) unter Verwendung von EDCI und HOBt in Gegenwart von Trimethylamin und Dichlormethan-Lösemittel erhalten. ¹H NMR (DMSO-d6, 400MHz): δ 10,97(1H, s), 8,98(1H, d, J=2,0Hz), 8,23(1H, dd, J=2,0 und 4,0Hz), 7,94(1H, d, J=8,0Hz), 7,74(1H, d, J=8,0Hz), 7,39(1H, dd, J=2,0 und 8,0Hz), 6,99(2H, m), 4,59(2H, d, J=12,0Hz), 3,22(2H, m), 2,94(1H, m), 2,08(2H, d, J=12,0Hz), 1,70(2H, m)
12. 12. N-([1,2,4]Triazol[4,3-alpyridin-3-yl)-1-(2-cyano-4-nitrophenyl) piperidin-4-carboxamid (Verbindung 12)
    
    Wie im oben stehenden Reaktionsschema 12 gezeigt, wurden 20 mg von Verbindung 11 (3) mit einer Ausbeute von etwa 23% durch Amidkopplungsreaktion von Ausgangsmaterial (1) und Carbonsäure (2) unter Verwendung von EDCI und HOBt in Gegenwart von Trimethylamin und Dichlormethan-Lösemittel erhalten. ¹H NMR (DMSO-d6, 400 MHz): δ 11,00 (1H, s), 8,54 (1H, d, J = 4,0Hz), 8,3 (1H, dd, J = 10,0 und 2,0Hz), 7,96 (1H, d, J=4,0Hz), 7,75 (1H, d, J = 8,0Hz), 7,39 (1H, ddd, J=6,6, 4,0 und 1,4Hz), 7,30 (1H, d, J = 12,0Hz), 6,98 (1H, t, J= 6,0Hz), 4,05 (2H, d, J=12,0Hz), 3,25 (2H, t, J = 12,0Hz), 2,87 (1H, m), 2,13 (2H, d, J = 12,0Hz), 1,885 (2H, d, J = 12,0Hz)

Triazole pyridine-based derivative compounds 1 to 12 of chemical formulas 1 to 12 were synthesized as follows:

1. 1. N - ([1,2,41Triazole [4,3-alpyridin-3-vl) -1- (2-cyanophenyl) piperidine-4-carboxamide (compound 1 )
   
   As shown in Reaction Scheme 1 above, 21 mg of compound 1 (3) obtained in about 25% yield by amide coupling reaction of a starting material (1) and carboxylic acid (2) using EDCI and HOBt in the presence of trimethylamine and dichloromethane solvent. ¹ H NMR (DMSO-d6, 400 MHz): δ 10.96 (1H, s), 7.96 (1H, d, J = 8.0Hz), 7.73 (1H, m), 7.61 ( 1H, t, J = 8.0Hz), 7.4 (1H, ddd, J = 12.0, 4.0 and 0.8Hz), 7.22 (1H, d, J = 8.0Hz), 7 , 11 (1H, t, J = 8.0Hz), 6.99 (1H, t, J = 6.0Hz), 3.58 (2H, d, J = 12.0Hz), 2.91 (2H, t, J = 12.0Hz), 2.50 (1H, m) 2.10 (2H, d, J = 12.0Hz), 1.91 (2H, t, J = 6.0Hz)
2. 2. N - ([1,2,41Triazol [4,3-alpyridin-3-yl) -1- (2-nitrophenyl) piperidine-4-carboxamide (compound 2nd )
   
   As shown in Reaction Scheme 2 above, 25 mg of Compound 2 (3) was obtained in approximately 29% yield by amide coupling reaction of a starting material (1) and carboxylic acid (2) using EDCI and HOBt in the presence of trimethylamine and dichloromethane Obtain solvent. ¹ H NMR (DMSO-d6, 400 MHz): δ 10.95 (1H, s), 7.95 (1H, d, J = 8.0 Hz), 7.81 (1H, dd, J = 8.0) and 4.0Hz), 7.75 (1H, d, J = 8.0Hz), 7.59 (1H, td, J = 8.0, and 4.0Hz), 7.38 (2H, m), 7.13 (1H, t, J = 8.0Hz), 7.00 (1H, t, J = 4.0Hz), 3.27 (2H, d, J = 12.0Hz), 2.91 (2H , t, J = 12.0Hz), 2.71 (1H, m), 2.03 (2H, d, J = 12.0Hz), 1.85 (2H, m)
3. 3. N - ([1,2,41Triazol [4,3-alpyridin-3-yl) -1-phenylpiperidine-4-carboxamide (Compound 3)
   
   As shown in Reaction Scheme 3 above, 24 mg of Compound 3 (3) was obtained in about 28% yield by amide coupling reaction of a starting material (1) and carboxylic acid (2) using EDCI and HOBt in the presence of trimethylamine and dichloromethane -Solvent obtained. ¹ H NMR (CDCl ₃ -d, 400 MHz): δ 11.65 (1H, s), 7.91 (1H, d, J = 4.0Hz), 7.67 (1H, d, J = 8, 0Hz), 7.27 (3H, m), 6.97 (2H, d, J = 8.0Hz), 6.9 (2H, m), 3.76 (2H, dt, J = 11.0 and 3.1 Hz), 2.9 (3H, td, J = 10.0 and 3.2Hz), 2.19 (2H, dd, J = 14.0 and 2.0Hz), 2.07 (2H, ddd, J = 24.0, 12.0 and 4.0Hz)
4. 4. N - ([1,2,41Triazol [4,3-alpyridin-3-yl) -1- (4-nitrophenyl) piperidine-4-carboxamide (Compound 4)
   
   As shown in Reaction Scheme 4 above, 22 mg of compound 4 (3) was obtained in about 25% yield by amide coupling reaction of a starting material (1) and carboxylic acid (2) using EDCI and HOBt in the presence of trimethylamine and dichloromethane Obtain solvent. ¹ H NMR (DMSO-d6, 400 MHz): δ 10.96 (1H, s), 8.06 (2H, d, J = 8.0Hz), 7.93 (1H, d, J = 4.0Hz ), 7.74 (1H, d, J = 8.0Hz), 7.39 (1H, ddd, J = 8.4, 6.6 and 1.8Hz), 7.07 (2H, d, J = 8.0Hz), 6.97 (1H, t, J = 6.0Hz), 4.13 (2H, d, J = 12.0Hz), 3.14 (2H, t, J = 12.0Hz), 2.89 (1H, m), 2.05 (2H, d, J = 12.0Hz), 1.75 (2H, m)
5. 5. N - ([1,2,41Triazol [4,3-alpyridin-3-yl) -1- (2-nitro-4- (trifluoromethyl) phenyl) piperidine-4-carboxamide (Compound 5)
   
   As shown in Reaction Scheme 5 above, 28 mg of Compound 5 (3) was obtained in about 27% yield by amide coupling reaction of a starting material (1) and carboxylic acid (2) using EDCI and HOBt in the presence of trimethylamine and dichloromethane Obtain solvent. ¹ H NMR (DMSO-d6, 400 MHz): δ 10.97 (1H, s), 8.16 (1H, d, J = 4.0Hz), 7.87 (1H, d, J = 4.0Hz ), 7.85 (1H, d, J = 2.0Hz), 7.75 (1H, dt, J = 10.6 and 0.4Hz), 7.49 (1H, d, J = 8.0Hz) , 7.40 (1H, ddd, J = 10.2, 6.0 and 1.4Hz), 6.99 (1H, td, J = 7.5, 7.5 and 3.2Hz), 3.43 (2H, d, J = 16.0Hz), 3.07 (2H, t, J = 12.0Hz), 2.79 (1H, m), 2.05 (2H, d, J = 12.0Hz) , 1.85 (2H, t, J = 12.0Hz)
6. 6. N - ([1,2,41Triazol [4,3-alpyridin-3-yl) -1- (4- (trifluoromethvl) phenyl) piperidine-4-carboxamide (Compound 6)
   
   As shown in Reaction Scheme 6 above, 24 mg of Compound 6 (3) was obtained in about 27% yield by amide coupling reaction of a starting material (1) and carboxylic acid (2) using EDCI and HOBt in the presence of trimethylamine and dichloromethane Obtain solvent. ¹ H NMR (DMSO-d6, 400 MHz): δ 10.94 (1H, s), 7.93 (1H, d, J = 8.0Hz), 7.74 (1H, d, J = 12.0Hz ), 7.50 (2H, d, J = 12.0Hz), 7.39 (1H, t, J = 8.0Hz), 7.10 (2H, d, J = 8.0Hz), 6.97 (1H, t, J = 6.0Hz), 3.97 (2H, d, J = 16.0Hz), 2.95 (2H, t, J = 12.0Hz), 2.81 (1H, m) , 2.02 (2H, d, J = 12.0Hz), 1.76 (2H, m)
7. 7. N - ([1,2,41Triazole [4,3-alpyridin-3-y1) -1- (4-fluoro-2-nitrophenyl) piperidine-4-carboxamide (Compound 7)
   
   As shown in Reaction Scheme 7 above, 28 mg of Compound 7 (3) was obtained in about 27% yield by amide coupling reaction of a starting material (1) and carboxylic acid (2) using EDCI and HOBt in the presence of trimethylamine and dichloromethane Obtain solvent. ¹ H NMR (DMSO-d6, 400 MHz): δ 10.94 (1H, s), 7.94 (1H, d, J = 4.0Hz), 7.83 (1H, dd, J = 8.0) and 4.0Hz), 7.75 (1H, d, J = 8.0Hz), 7.52 (2H, m), 7.40 (1H, dd, J = 12.0 and 6.0Hz), 6 , 99 (1H, t, J = 8.0Hz), 3.20 (2H, d, J = 12.0Hz), 2.88 (2H, t, J = 12.0Hz), 2.69 (1H, m), 2.03 (2H, d, J = 8.0Hz), 1.82 (2H, m)
8. 8. N - ([1,2,41Triazol [4,3-alpyridin-3-yl) -1- (4-acetylphenyl) piperidine-4-carboxamide (Compound 8)
   
   As shown in Reaction Scheme 8 above, 24 mg of compound 8 (3) was obtained in approximately 28% yield by amide coupling reaction of a starting material (1) and carboxylic acid (2) using EDCI and HOBt in the presence of trimethylamine and dichloromethane Obtain solvent. ¹ H NMR (DMSO-d6, 400 MHz): δ 10.95 (1H, s), 7.93 (1H, d, J = 8.0Hz), 7.81 (2H, d, J = 8.0Hz ), 7.74 (1H, d, J = 12.0Hz), 7.39 (1H, ddd, J = 8.4, 6.4 and 1.6Hz), 6.99 (3H, m), 4 , 04 (2H, d, J = 12.0Hz), 3.00 (2H, t, J = 12.0Hz), 2.84 (1H, m), 2.45 (3H, s), 2.02 (2H, d, J = 12.0Hz), 1.74 (2H, m)
9. 9. N - ([1,2,4] triazole [4,3-alpyridin-3-yl) -1- (4-chloro-2-nitrophenyl) piperidine-4-carboxamide (compound 9)
   
   
   As shown in Reaction Scheme 9 above, 28 mg of Compound 9 (3) was obtained in about 26% yield by amide coupling reaction of a starting material (1) and carboxylic acid (2) using EDCI and HOBt in the presence of trimethylamine and dichloromethane Obtain solvent. ¹ H NMR (DMSO-d6, 400 MHz): δ 10.95 (1H, s), 7.95 (2H, m), 7.75 (1H, d, J = 8.0Hz), 7.65 ( 1H, dd, J = 12.0 and 8.0Hz), 7.39 (2H, m), 6.99 (1H, t, J = 8.0Hz), 3.26 (2H, d, J = 12 , 0Hz), 2.92 (2H, t, J = 12.0Hz), 2.72 (1H, m), 2.03 (2H, d, J = 8.0Hz), 1.83 (2H, m )
10. 10. N - ([1,2,4] triazole [4,3-alpyridin-3-yl) -1- (4-cyanophenyl) piperidine-4-carboxamide (compound 10)
    
    As shown in Reaction Scheme 10 above, 20 mg of compound 10 (3) was obtained in about 23% yield by amide coupling reaction of starting material (1) and carboxylic acid (2) using EDCI and HOBt in the presence of trimethylamine and dichloromethane solvent receive. ¹ H NMR (DMSO-d6, 400MHz): δ 10.94 (1H, s), 7.93 (1H, d, J = 8.0Hz), 7.74 (1H, d, J = 8.0Hz) , 7.58 (1H, d, J = 8.0Hz), 7.39 (1H, dd, J = 8.0 and 8.0Hz), 7.06 (2H, d, J = 8.0Hz), 6.97 (1H, m), 4.03 (2H, d, J = 12.0Hz), 3.01 (2H, m), 2.83 (1H, m), 2.02 (2H, d, J = 16Hz), 1.73 (2H, m)
11. 11. N - ([1,2,41Triazol [4,3-alpyridin-3-yl) -1- (5-nitropyridin-2-yl) piperidine-4-carboxamide (Compound 11)
    
    As shown in Reaction Scheme 11 above, 20 mg of Compound 11 (5) was obtained in approximately 26% yield by amide coupling reaction of starting material (1) and carboxylic acid (2) using EDCI and HOBt in the presence of trimethylamine and dichloromethane solvent receive. ¹ H NMR (DMSO-d6, 400MHz): δ 10.97 (1H, s), 8.98 (1H, d, J = 2.0Hz), 8.23 (1H, dd, J = 2.0 and 4.0Hz), 7.94 (1H, d, J = 8.0Hz), 7.74 (1H, d, J = 8.0Hz), 7.39 (1H, dd, J = 2.0 and 8 , 0Hz), 6.99 (2H, m), 4.59 (2H, d, J = 12.0Hz), 3.22 (2H, m), 2.94 (1H, m), 2.08 ( 2H, d, J = 12.0Hz), 1.70 (2H, m)
12. 12. N - ([1,2,4 ] triazole [4,3-alpyridin-3-yl) -1- (2-cyano-4-nitrophenyl) piperidine-4-carboxamide (compound 12)
    
    As shown in Reaction Scheme 12 above, 20 mg of Compound 11 (3) was obtained in about 23% yield by amide coupling reaction of starting material (1) and carboxylic acid (2) using EDCI and HOBt in the presence of trimethylamine and dichloromethane solvent receive. ¹ H NMR (DMSO-d6, 400 MHz): δ 11.00 (1H, s), 8.54 (1H, d, J = 4.0Hz), 8.3 (1H, dd, J = 10.0 and 2.0Hz), 7.96 (1H, d, J = 4.0Hz), 7.75 (1H, d, J = 8.0Hz), 7.39 (1H, ddd, J = 6.6, 4.0 and 1.4Hz), 7.30 (1H, d, J = 12.0Hz), 6.98 (1H, t, J = 6.0Hz), 4.05 (2H, d, J = 12 , 0Hz), 3.25 (2H, t, J = 12.0Hz), 2.87 (1H, m), 2.13 (2H, d, J = 12.0Hz), 1.885 (2H, d, J = 12.0Hz)

<Example 1> Confirmation of an inhibitory effect of Tankyrase (TNKS) enzyme activity

Whether the triazole pyridine-based derivative compounds 1 to 12 can inhibit the activity of tankyrase (TNKS) enzyme was determined using the tankyrase-1 colorimetric activity assay kit (Cat # 4500-192-K, Trevigen, RnD systems, USA) determined by the manufacturer and {2- (4- (trifluoromethylphenyl) -3,5,7,8-tetradihydro-thiopyrano [4,3-d] pyrimidin-4-one (XAV939) as a positive control.

In addition, it was confirmed by measurement with PARP-1 colorimetric assay kit (Cat # 4677-096-K, Trevigen, RnD Systems) according to the manufacturer's instructions and with olaparib 1µM as a positive control whether compound 1 to compound 12 PARP- (poly -ADP ribose polymerase) can inhibit enzyme activity.

As a result, TNKS and PARP inhibitory activity of Compounds 1 to 12 was confirmed as shown in Table 1. [Table 1] TNKS-1 inhibitory activity (compared to control 0%) PARP inhibitory activity Concentration (µM) 10th 10th Triazole pyridine compound 1 95.2 ± 0.5 0 Triazole pyridine compound 2 94.3 ± 1.3 0 Triazole pyridine compound 3 58.7 ± 2.4 0 Triazole pyridine compound 4 30.8 ± 4.7 1.5 ± 0.8 Triazole pyridine compound 5 70.4 ± 2.5 0 Triazole pyridine compound 6 51.7 ± 1.7 1.2 ± 0.4 Triazole pyridine compound 7 75.4 ± 1.2 0 Triazole pyridine compound 8 65.8 ± 2.3 0.8 ± 0.2 Triazole pyridine compound 9 72.9 ± 4.7 1.1 ± 0.5 Triazole pyridine compound 10 59.6 ± 5.2 0.8 ± 0.4 Triazole pyridine compound 11 47.4 ± 3.4 0.6 ± 0.1 Triazole pyridine compound 12 94.8 ± 0.8 0

The control group XAV939 (10µM) inhibited TNKS-1 enzyme activity by about 96%, olaparib (10µM) inhibited PARP enzyme activity by 98%. On the other hand, triazole pyridine-based compounds 1 to 12 showed the inhibition of TNKS-1 enzyme activity as shown in Table 1 while the PARP-1 activity was not inhibited.

From the above results, it was confirmed that the activity of the triazole pyridine-based derivatives was specific for the TNKS-1 enzyme.

In addition, 50% inhibitory concentration of TNKS-1 activity was shown in Table 2 in relation to compound 1 and connection 2nd confirms which ones have the best activity. [Table 2] TNKS-1 activity 50% inhibitory concentration TNKS-1 (µM) (IC ₅₀ ) Triazole pyridine compound 1 0.760 Triazole pyridine compound 2 0.863

<Example 2> Confirmation of inhibitory effect on β-catenin expression

Cell preparation

DLD-1, SW480 and SKMEL28 cell lines were purchased from ATCC (American Type Culture Collection). DLD-1 and SW480 cells were incubated in DMEM medium (Dulbecco's Modified Eagle's Medium) and SKMEL28 cells were in MEM medium, 10% fetal bovine serum (FBS) and 100 µg / ml streptomycin and 100 unit / ml, respectively Penicillin were added, incubated under 5% CO ₂ and 37 ° C.

The Vemurafenib-resistant SKMEL28 cell line was continuously treated with 5 µM vemurafenib for 1 month, followed by treatment with Vemurafenib using a cell line with 40% inhibition of cell proliferation.

Confirmation of inhibition of β-catenin expression

Western blot confirmed whether the triazole pyridine-based compound 1 and connection 2nd Inhibit β-catenin protein expression by TNKS inhibition and maintain AXIN2 protein.

First, ALD-mutated DLD-1 colon cancer cells were each linked with 10 µM of compound 1 or connection 2nd treated and then cultured under 5% CO ₂ and 37 ° C for 24 hours.

After that, the cells that were connected 1 or connection 2nd treated, obtained and cytoplasmic fractions were obtained by methods commonly used in the art.

The fractions were separated by electrophoresis with a 10% gradient SDS-PAGE and transferred to nitrocellulose membrane (BioRad, Hercules, CA, USA). The membrane was blocked with 5% skim milk and then reacted with anti-β-catenin (Cell signaling, USA) and anti-AXIN2 antibody (Cell signaling, USA) as a primary antibody, followed by incubation of horseradish peroxidase-conjugated Anti-mouse IgG antibody (Santa Cruz Biotechnology, USA), and visualized with an ECL system (GE, USA).

As a result, as in 2nd shown was expression of β-catenin protein by the compounds 1 and 2nd decreased, which are TNKS inhibitors, while the AXIN2 protein was increased.

Confirmation of inhibition of β-catenin subgene expression

DLD-1 colon cancer cells mutated with the APC gene were divided into 6 cm plates with a number of 5 × 10 ⁵ and with 10 μM compound 1 or connection 2nd treated and cultivated for 8 hours under the conditions of 5% CO ₂ and 37 ° C.

After receiving cells that are connected 1 or connection 2nd total RNA was extracted using RNeasy Kit (Qiagen, USA) and cDNA was synthesized using M-MLV reverse transcriptase (Enzynomics, Korea). Oligomers capable of binding β-catenin subgenes VEGF2 and Birc5 / Survivin were synthesized in Cosmogentec (Korea), labeled with a SYBR-Green / Fluorescein qPCR master mix (Thermo Scientific, USA) and real-time PCR was performed in one Lightcycler 96 real-time PCR system (RocheDiagnositics, Germany) measured according to the manufacturer's guidelines.

As a result, it was confirmed that VEGF2 and Birc5 / survivin expression were associated with 1 or connection 2nd treated cells is significantly reduced, as in 3rd shown.

<Example 3> Confirmation of anti-cancer effects by combined radiation treatment

Cultivated DLD-1 colon cancer cells were placed in a 96 W cell culture plate in the number of 3 × 10 ³ , with compound 1 and connection 2nd treated at a concentration of 20 µM and irradiated with 3 Gy gamma radiation and incubated for 3 days under conditions of CO ₂ and 37 ° C.

Then 0.5 mg / mL MTT- (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) solution was added to each well and incubated for a further 3 hours at 37 ° C. After incubation, the supernatant was removed and the formazane crystals formed were dissolved in DMSO and measured at 590 nm using a spectrometer (Labsystems, USA). The control group was treated with DMSO and cell viability was reported as a relative growth rate relative to 100%.

As a result, as in 4th It was confirmed that the combined treatment of the triazole pyridine-based compound 1 or connection 2nd at a concentration of 20 µM with the radiation, the viability decreased.

The results above confirmed that the triazole pyridine-based compound 1 and connection 2nd can be used as a radiation therapy adjuvant, which can enhance the effect in radiation therapy.

<Example 4> Confirmation of anti-cancer effects by combined treatment of anti-cancer drugs

DLD-1 cells and SW480 cells cultured in the same manner as in Example 3 were mixed with 20 µM of compound 1 or connection 2nd in combination with 5-FU (10 µM), a colon cancer anticancer agent, treated for 48 hours and then subjected to an MTT assay for cell viability analysis.

As a result, as in 5 was shown to inhibit cell proliferation in the experimental group, in the 5-FU and compound 1 or connection 2nd were co-administered 2-fold and 1.5-fold higher than that of the control in which 5-FU, an anti-cancer agent, was administered alone.

The above results confirm that the triazole pyridine-based compound can be used as an anti-cancer adjuvant capable of enhancing effects of an anti-cancer drug.

<Example 5> Confirmation of anti-cancer activity in resistant cancer cells by inhibiting tumor factor YAP activity

Confirmation of an inhibitory effect on YAP / TAZ signaling

Since TNKS was shown to activate tumor factor YAP by angiomotin, real-time PCR was carried out in the same manner as in Example 2 to confirm the inhibitory effect on YAP / TAZ signaling by triazole pyridine compound, which is the TNKS inhibitor, and the expression values of YAP / TAZ subgenes CYR61 and CTGF have been confirmed.

As a result, as in 6 It was confirmed that expression of CYR61 and CTGF in the cells associated with compound 1 or connection 2nd had been treated was significantly reduced.

The results above confirm that the triazole pyridine compound can be directed to the YAP / TAZ tumor factor.

Confirmation of anti-cancer effects by combined treatment with anti-cancer agents in resistant cancer cells

The B-RAF inhibitory anti-cancer drug, vemurafenib (PLX4032), has been reported to be involved in YAP / TAZ signaling (EMBO J. 2016; 35 (5): 462), and SKMEL28 melanoma cells and vemurafenib-resistant SKMEL28- Cells that had been cultured in previous experiments with compound 1 treated at concentrations of 10 µM, 20 µM and 30 µM and 10 µM or 20 µM and cell viability was analyzed by MTT assay.

As a result, as in 7 shown, the cell proliferation rate in the SKMEL28 cell line and the vemurafenib-resistant SKMEL28 cells was dependent on the concentration of compound 1 decreased. In particular, it was confirmed that cell proliferation was reduced more in vemurafenib-resistant SKMEL28 cells than in SKMEL28 cells, and therefore the triazole pyridine compound of the present invention can be used as a very effective anti-cancer agent.

Although the present invention has been described in particular with reference to its specific embodiments, it will be apparent to those skilled in the art that this specific description is only a preferred embodiment and that the scope of the present invention is not limited thereby, that is, the practical scope of the present invention defined by the appended claims and their equivalents.

Claims (15)

Triazole pyridine-based derivative represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof:

in chemical formula 1, each of R1 and R2 can be the same or different and selected from hydrogen, CN, NO2, CF3, halogen and acetyl, and X is arbitrarily selected from nitrogen and carbon. Triazole pyridine-based derivative or a pharmaceutically acceptable salt thereof Claim 1 wherein each of R ¹ and R ^{2 may be the} same or different and selected from the group consisting of CN, NO ₂ , CF ₃ and halogen, and X is carbon. Triazole pyridine-based derivative or a pharmaceutically acceptable salt thereof Claim 1 , wherein the triazole pyridine-based derivative is selected from the group consisting of N - ([1,2,4,] triazole [4,3-a] pyridin-3-yl) -1- (2-cyanophenyl) piperidin-4 -carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (2-nitrophenyl) piperidine-4-carboxamide, N - ([1,2,4 ] Triazole [4,3-a] pyridin-3-yl) -1-phenylpiperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1 - (4-nitrophenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (2-nitro-4- (trifluoromethyl) phenyl ) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (4- (trifluoromethyl) phenyl) piperidine-4-carboxamide, N- ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (4-fluoro-2-nitrophenyl) piperidine-4-carboxamide, N - ([1,2,4] Triazole [4,3-a] pyridin-3-yl) -1- (4-acetylphenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridine-3- yl) -1- (4-chloro-2-nitrophenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridine- 3-yl) -1- (4-cyanophenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (5-nitropyridine- 2-yl) piperidine-4-carboxamide and N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (2-cyano-4-nitrophenyl) piperidine-4- carboxamide. A pharmaceutical composition for preventing or treating cancer, comprising a triazole pyridine-based derivative represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, as an active ingredient:

in chemical formula 1, each of R1 and R2 can be the same or different and selected from hydrogen, CN, NO2, CF3, halogen and acetyl, and X is arbitrarily selected from nitrogen and carbon. Pharmaceutical composition for the prevention or treatment of cancer after Claim 4 wherein each of R ¹ and R ^{2 can be the} same or different and selected from the group consisting of CN, NO ₂ , CF ₃ and halogen, and X is carbon. Pharmaceutical composition for the prevention or treatment of cancer after Claim 4 , wherein the triazole pyridine-based derivative is selected from the group consisting of N - ([1,2,4,] triazole [4,3-a] pyridin-3-yl) -1- (2-cyanophenyl) piperidin-4 -carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (2-nitrophenyl) piperidine-4-carboxamide, N - ([1,2,4 ] Triazole [4,3-a] pyridin-3-yl) -1-phenylpiperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1 - (4-nitrophenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (2-nitro-4- (trifluoromethyl) phenyl ) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (4- (trifluoromethyl) phenyl) piperidine-4-carboxamide, N- ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (4-fluoro-2-nitrophenyl) piperidine-4-carboxamide, N - ([1,2,4] Triazole [4,3-a] pyridin-3-yl) -1- (4-acetylphenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridine-3- yl) -1- (4-chloro-2-nitrophenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (4- cyanophenyl) piperidine-4-carboxamide, N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (5-nitropyridin-2-yl) piperidine-4-carboxam id and N - ([1,2,4] triazole [4,3-a] pyridin-3-yl) -1- (2-cyano-4-nitrophenyl) piperidine-4-carboxam id. Pharmaceutical composition for the prevention or treatment of cancer after Claim 4 , wherein the triazole pyridine-based derivative represented by chemical formula 1 inhibits activity of tankyrase (TNKS) enzyme. Pharmaceutical composition for the prevention or treatment of cancer after Claim 4 , the cancer being one of the group consisting of lung cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, breast cancer, stomach cancer, liver cancer, colon cancer, skin cancer, head or neck cancer, uterine cancer, ovarian cancer, Brain cancer, larynx cancer, prostate cancer, bladder cancer, esophageal cancer, thyroid cancer, kidney cancer, blood cancer and rectal cancer. A composition for enhancing an anti-cancer activity, comprising a triazole pyridine-based derivative represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, as an active ingredient:

in Chemical Formula 1, each of R1 and R2 can be the same or different and selected from hydrogen, CN, NO2, CF3, halogen and acetyl, and X is arbitrarily selected from nitrogen and carbon. Composition to increase anti-cancer effects after Claim 9 , the composition being treated in combination with an anti-cancer agent or radiation to increase anti-cancer activity. Composition to increase anti-cancer effects after Claim 10 , wherein the anti-cancer agent is one or more selected from the group consisting of cisplatin, 5-fluorouracil, paclitaxel, doxorubicin, daunorubicin, vinblastine, vincristine, actinomycin D, teniposide, etoposide, cyclophosphamide, epirubicin, adriamycin, daunomycin and mitcin. Composition to increase anti-cancer effects after Claim 9 , wherein the composition comprises 1 to 99 parts by weight of an anti-cancer agent and 1 to 99 parts by weight of a triazolopyridine derivative represented by Chemical Formula 1 based on 100 parts by weight of the total composition. Composition to increase anti-cancer effects after Claim 9 , the cancer being one selected from the group consisting of lung cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, breast cancer, stomach cancer, liver cancer, colon cancer, skin cancer, head or neck cancer, uterine cancer, ovarian cancer, Brain cancer, larynx cancer, prostate cancer, bladder cancer, esophageal cancer, thyroid cancer, kidney cancer, blood cancer and rectal cancer. A healthy food for preventing or improving cancer, comprising a triazolopyridine-based derivative represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, as an active ingredient:

in chemical formula 1, each of R1 and R2 can be the same or different and selected from hydrogen, CN, NO ₂ , CF ₃ , halogen and acetyl, and X is arbitrarily selected from nitrogen and carbon. Healthy food to prevent or improve cancer after Claim 14 , the cancer being one selected from the group consisting of lung cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, breast cancer, stomach cancer, liver cancer, colon cancer, skin cancer, head or neck cancer, uterine cancer, Ovarian cancer, brain cancer, larynx cancer, prostate cancer, bladder cancer, esophageal cancer, thyroid cancer, kidney cancer, blood cancer and rectal cancer.

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