Composition for preventing or treating cancer disease comprising aryl piperidine derivative

Abstract

The present invention relates to a pharmaceutical composition comprising an aryl piperidine derivative compound or a pharmaceutically acceptable salt thereof for use in the treatment or prevention of tankyrase-related cancer diseases. The pharmaceutical composition inhibits the enzymatic activity of tankyrase (TNKS), inhibits the expression of a beta-catenin target gene and the protein expression of active beta-catenin, stabilizes AXIN2 protein levels, and exhibits anti-cancer activity in a cancer cell line, so that the compound can be used as an anti-cancer agent, in particular as an anti-cancer agent. And elicits synergistic effects in combination therapy with other anti-cancer agents.

Description

Technical Field

[0001] This disclosure provides compositions for the prevention or treatment of cancer, the compositions comprising arylpiperidine derivatives. Background Technology

[0002] Thanks to advancements in modern medicine, many diseases are being treated and prevented, but cancer remains one of the most difficult diseases to treat. Currently, cancer is the leading cause of death, and its incidence continues to rise.

[0003] Cancer treatments include chemotherapy, surgery, and radiation therapy. Chemotherapy is the most common method of treating cancer using anticancer drugs. Currently, approximately 60 different types of anticancer drugs are used clinically, and new anticancer drugs are constantly being developed as knowledge about cancer development and the characteristics of cancer cells accumulates. However, most anticancer drugs used in current clinical practice frequently cause side effects such as nausea, vomiting, oral and small intestinal ulcers, diarrhea, hair loss, and bone marrow suppression that reduces the production of effective blood components. For example, mitomycin C is known to have side effects such as kidney failure, while doxorubicin can cause bone marrow suppression.

[0004] Meanwhile, telomerases (TNKS) are regulators of telomerase activity and are known to participate in DNA damage responses and Wnt signaling. This family of telomerase proteins consists of telomerase 1 (TNKS1) and telomerase 2 (TNKS2), which share 85% amino acid identity.

[0005] Anchored polymerase 1 / 2 is known to be a regulator of the WNT / β-catenin signaling pathway through interaction with AXIN protein, and a regulator of the hippo signaling pathway through interaction with members of the AMOT protein family. Inhibition of anchored polymerase 1 / 2 is known to suppress YAP oncogenic function by increasing AXIN protein levels (even in the absence of dysfunctional and truncated forms of APC protein), decreasing cellular β-catenin levels, and stabilizing the AMOT protein family.

[0006] To date, there are no TNKS inhibitors available for clinical use, and given the critical importance of WNT / β-catenin signaling and hippo signaling in various cancers, there is an urgent need to develop anticancer agents that utilize TNKS inhibitors. Summary of the Invention

[0007] Technical issues

[0008] The purpose of this disclosure is to provide arylpiperidine derivatives or pharmaceutically acceptable salts thereof, and pharmaceutical compositions comprising the thereof for the treatment or prevention of end-anchor polymerase-related cancer diseases.

[0009] Another object of this disclosure is to provide a pharmaceutical composition for combination therapy of end-anchor polymerase-related cancers, said pharmaceutical composition comprising an arylpiperidine derivative compound or a pharmaceutically acceptable salt thereof and an anticancer agent.

[0010] Another object of this disclosure is to provide a pharmaceutical composition for enhancing anticancer effects against end-anchor polymerase-related cancers, said pharmaceutical composition comprising an arylpiperidine derivative compound or a pharmaceutically acceptable salt thereof, and an anticancer agent.

[0011] Another object of this disclosure is to provide a health-functional food composition for improving or preventing end-anchor polymerase-related cancers, the health-functional food composition comprising an arylpiperidine derivative compound or a pharmaceutically acceptable salt thereof, and an anticancer agent.

[0012] Technical solution

[0013] To achieve the above objectives, this disclosure provides arylpiperidine derivatives or pharmaceutically acceptable salts thereof as shown in the following chemical formula 1.

[0014] [Chemical Formula 1]

[0015]

[0016] In the chemical formula 1, R 1 To R 3 They can be the same or different, and each can be independently selected from hydrogen (H), nitro (NO2), and cyano (CN); R 4 It can be selected from or Any of the following; E1 to E8 may be the same or different, and each may be CH or N independently; R' may be hydrogen (H) or (C1~C4) alkyl, and n may be any one of 1 to 5.

[0017] In addition, this disclosure provides arylpiperidine derivatives or pharmaceutically acceptable salts thereof as shown in the following chemical formula 2.

[0018] [Chemical Formula 2]

[0019]

[0020] In the chemical formula 2, R 1 To R 3 They can be the same or different, and each can be independently selected from hydrogen (H) or cyano (CN); R 4 It can be or E1 to E8 can be the same or different, and each can be CH or N independently; L can be NR' or NH2. + Furthermore, R' can be a (C1~C4) alkyl or a (C1~C5) alkoxycarbonyl.

[0021] In addition, this disclosure provides pharmaceutical compositions for treating or preventing end-anchor polymerase-related cancers, the pharmaceutical compositions comprising the arylpiperidine derivative compound or a pharmaceutically acceptable salt thereof.

[0022] Furthermore, this disclosure provides pharmaceutical compositions for combination therapies for treating end-anchor polymerase-related cancers, the pharmaceutical compositions comprising the arylpiperidine derivative compound or a pharmaceutically acceptable salt thereof, and an anticancer agent.

[0023] Furthermore, this disclosure provides pharmaceutical compositions for enhancing anticancer effects against end-anchor polymerase-related cancers, the pharmaceutical compositions comprising the arylpiperidine derivative compound or a pharmaceutically acceptable salt thereof, and an anticancer agent.

[0024] Furthermore, this disclosure provides a health-promoting functional food composition for improving or preventing end-anchor polymerase-related cancers, the health-promoting functional food composition comprising the arylpiperidine derivative compound or a pharmaceutically acceptable salt thereof, and an anticancer agent.

[0025] Beneficial effects

[0026] This disclosure relates to arylpiperidine-derived compounds or pharmaceutically acceptable salts thereof, which inhibit the enzymatic activity of terminal anchor polymerase (TNKS), suppress the expression of β-catenin target genes and the protein expression of active β-catenin, stabilize AXIN2 protein levels, and exhibit anticancer activity in cancer cell lines, thereby enabling the compounds to be used as anticancer agents; and due to their synergistic effect in anticancer activity when used in combination with fluorouracil (5-FU), they can be used as combination therapy with conventional anticancer agents. Attached Figure Description

[0027] Figure 1 A schematic diagram of the synthesis of arylpiperidine derivatives is shown.

[0028] Figure 2The results of the identification of the inhibitory effect on the expression of active β-catenin and the results of the identification of the stability of AXIN2 protein expression after treatment with 10 μM of N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-2-(1-(2-cyanophenyl)piperidin-4-yl)acetamide (hereinafter referred to as TI-61815) and N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(2-cyanophenyl)piperidin-4-yl)propenylamine (hereinafter referred to as TI-61910) of two types of human colon cancer cell lines COLO320DM and SW403 are shown.

[0029] Figure 3 The results show the identification of the inhibitory effect on the subgene expression of β-catenin after treatment of the human colon cancer cell line COLO320DM with 10 μM of compounds TI-61815 and TI-61910 according to this disclosure.

[0030] Figure 4 The results of cell viability assessment by treating the human colon cancer cell line COLO320DM with different concentrations of compounds TI-61815 and TI-61910 according to this disclosure are shown.

[0031] Figure 5 The results of cell viability assessment in the human colon cancer cell line COLO320DM by co-treatment with the anticancer agent (5-FU) using compounds TI-61815 and TI-61910 according to this disclosure are shown. Detailed Implementation

[0032] The following will describe this disclosure in more detail.

[0033] In researching and developing anticancer agents using therapeutically targeted TNKS inhibitors (TNKS is known to be crucial for WNT / β-catenin signaling and hippo signaling in various cancers), the inventors of this disclosure completed this disclosure by discovering compounds that inhibit the enzymatic activity of TNKS, suppress the expression of target genes of β-catenin and the protein expression of active β-catenin, stabilize AXIN2 protein levels, and exhibit excellent anticancer activity in human colon cancer cell lines.

[0034] This disclosure provides arylpiperidine derivatives or pharmaceutically acceptable salts thereof, as shown in Chemical Formula 1 below.

[0035] [Chemical Formula 1]

[0036]

[0037] In the chemical formula 1, R 1 To R 3 They can be the same or different, and each can be independently selected from hydrogen (H), nitro (NO2), and cyano (CN), R 4 It can be selected from or Any of the following, E1 to E8 may be the same or different, and each is independently CH or N, R' may be hydrogen (H) or (C1 to C4) alkyl, and n may be any one of 1 to 5.

[0038] In the chemical formula 1, R 2 It can be hydrogen (H) or nitro (NO2), R 4 It can be , , ,and In any of the following, E1 can be N, E3 to E5 and E8 can be CH, R' can be hydrogen (H) or methyl, and n can be any one of 1 to 3.

[0039] The arylpiperidine derivatives may be selected from the group consisting of: N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-2-(1-(2-cyanophenyl)piperidin-4-yl)acetamide, 2-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-b]pyridazin-3-yl)acetamide, 2-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-a]pyridin-3-yl)acetamide, 2-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-a]pyridin-3-yl)acetamide, 2-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-a]pyridin-5-yl)acetyl Amines, N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(2-cyanophenyl)piperidin-4-yl)propionamide, N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-4-(1-(2-cyanophenyl)piperidin-4-yl)butyramide, 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-b]pyridazin-3-yl)propionamide, 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-a]pyridin-3-yl)propionamide, 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-a]pyridin-3-yl)propionamide, 3-(1-(2-cyanophenyl) Piperidin-4-yl)-N-(imidazo[1,2-a]pyridin-5-yl)propionamide, 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(1-methyl-1H-indol-6-yl)propionamide, 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(1H-indol-4-yl)propionamide, 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(1H-indol-7-yl)propionamide, N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(2-nitrophenyl)piperidin-4-yl)propionamide, N-([ [1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(3-nitrophenyl)piperidin-4-yl)propamide, N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(4-nitrophenyl)piperidin-4-yl)propamide, N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(4-cyanophenyl)piperidin-4-yl)propamide, and N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(2,4-dinitrophenyl)piperidin-4-yl)propamide.

[0040] In the chemical formula 1, R 1 It can be cyano (CN), R 2 and R 3 It can be hydrogen (H), R 4 It can be E1, E6 and E7 can be N, E2 to E5 and E8 can be CH, and n can be 1 or 2.

[0041] In addition, this disclosure provides arylpiperidine derivatives or pharmaceutically acceptable salts thereof as shown in the following chemical formula 2.

[0042] [Chemical Formula 2]

[0043]

[0044] In the chemical formula 2, R 1 To R 3 They can be the same or different, and each can be independently selected from hydrogen (H) or cyano (CN), R 4 It can be or E1 to E8 can be the same or different, and each can be CH or N independently. L can be NR' or NH2. + Furthermore, R' can be a (C1~C4) alkyl or a (C1~C5) alkoxycarbonyl.

[0045] In the chemical formula 2, R 1 It can be cyano (CN), R 2 and R 3 It can be hydrogen (H), R 4 It can be E1, E6 and E7 can be N, E2 to E5 and E8 can be CH, and R' can be (C1~C2) alkyl or (C3~C4) alkoxycarbonyl.

[0046] The arylpiperidine derivatives may be selected from the group consisting of: (2-([1,2,4]triazolo[4,3-a]pyridin-3-ylamino)-2-oxoethyl)(1-(2-cyanophenyl)piperidin-4-yl)carbamate tert-butyl ester, N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-2-((1-(2-cyanophenyl)piperidin-4-yl)amino)acetamide, and N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-2-((1-(2-cyanophenyl)piperidin-4-yl)(methyl)amino)acetamide.

[0047] The N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-2-((1-(2-cyanophenyl)piperidin-4-yl)amino)acetamide can form an ionic bond with any of the groups selected from the group consisting of: acetate (CH3COO) - ), trifluoroacetate (CF3COO) - ), chloride ions (Cl) - ), and methyl p-toluenesulfonate (CH3C6H4SO3) - (but not limited to this).

[0048] In addition, this disclosure provides pharmaceutical compositions for treating or preventing end-anchor polymerase-related cancers, the pharmaceutical compositions comprising the arylpiperidine derivative compound or a pharmaceutically acceptable salt thereof.

[0049] Furthermore, this disclosure provides pharmaceutical compositions for combination therapies for treating end-anchor polymerase-related cancers, the pharmaceutical compositions comprising the arylpiperidine derivative compound or a pharmaceutically acceptable salt thereof, and an anticancer agent.

[0050] The anticancer agent may be selected from, but is not limited to, the group consisting of: 7-ethyl-10-hydroxycamptothecin, 5-fluorouracil, cisplatin, paclitaxel, doxorubicin, daunorubicin, vincristine, actinomycin D, teniposide, etoposide, cyclophosphamide, epirubicin, adriamycin, doxorubicin, and mitomycin-C.

[0051] Furthermore, this disclosure provides pharmaceutical compositions for enhancing anticancer effects against end-anchor polymerase-related cancers, the pharmaceutical compositions comprising the arylpiperidine derivative compound or a pharmaceutically acceptable salt thereof, and an anticancer agent.

[0052] The cancer may be selected from the group consisting of: colon cancer, lung cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, breast cancer, stomach cancer, liver cancer, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, breast cancer, brain cancer, laryngeal cancer, prostate cancer, bladder cancer, esophageal cancer, thyroid cancer, kidney cancer, leukemia, and rectal cancer, but is not limited thereto.

[0053] In other embodiments of this disclosure, the pharmaceutical composition may further include one or more additives selected from the group consisting of suitable carriers, excipients, disintegrants, sweeteners, coating agents, leavening agents, lubricants, flow aids, flavoring agents, antioxidants, buffers, antibacterial agents, diluents, dispersants, surfactants, binders, and lubricants commonly used in the preparation of pharmaceutical compositions.

[0054] Specifically, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate / ester, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil can be used as the carrier, excipient, and diluent. Solid dosage forms for oral administration can include tablets, pills, acid preparations, granules, and capsules, and these solid dosage forms can be prepared by mixing at least one or more excipients (e.g., starch, calcium carbonate, sucrose or lactose, and gelatin) into the composition. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Oral liquid dosage forms can include suspensions, liquids, emulsions, and syrups, and can include various excipients such as humectants, sweeteners, flavorings, and preservatives, in addition to commonly used simple diluents (e.g., water and liquid paraffin). Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, and suppositories. For non-aqueous solvents and suspensions, propylene glycol, polyethylene glycol, vegetable oils (such as olive oil), and injectable esters (such as ethyl oleate) can be used. As for the base material of suppositories, Witepsol, polyethylene glycol (macrogol), Tween 61, cocoa butter, laurinfat, and glycerin gelatin can be used.

[0055] According to embodiments of this disclosure, the pharmaceutical composition can be administered to a subject in a conventional manner via intravenous, intra-arterial, intraperitoneal, intramuscular, intra-arterial, intraperitoneal, intrasternal, transdermal, intranasal, inhalation, local, rectal, oral, intraocular, or intradermal routes.

[0056] The dosage of the active ingredient according to this disclosure can vary depending on the subject's condition and weight, the type and severity of the disease, the dosage form of the drug, and the route and duration of administration, and can be appropriately selected by those skilled in the art. The daily dose can be from 0.01 mg / kg to 200 mg / kg, preferably from 0.1 mg / kg to 200 mg / kg, and more preferably from 0.1 mg / kg to 100 mg / kg. The administration can be performed once daily or divided into multiple doses, but the scope of this disclosure is not limited thereto.

[0057] Furthermore, this disclosure provides a health-promoting functional food composition for improving or preventing end-anchor polymerase-related cancers, the health-promoting functional food composition comprising the arylpiperidine derivative compound or a pharmaceutically acceptable salt thereof, and an anticancer agent.

[0058] The cancer may be selected from the group consisting of: colon cancer, lung cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, breast cancer, stomach cancer, liver cancer, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, breast cancer, brain cancer, laryngeal cancer, prostate cancer, bladder cancer, esophageal cancer, thyroid cancer, kidney cancer, leukemia, and rectal cancer, but is not limited thereto.

[0059] The aforementioned health functional foods may contain various nutritional supplements, vitamins, minerals (electrolytes), flavoring agents (such as synthetic and natural flavoring agents), coloring agents and thickeners (cheese, chocolate, etc.), pectic acid and its salts, alginate / esters and their salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonating agents used in carbonated beverages.

[0060] In addition, it may contain fruit pulp used in the production of natural fruit juices, synthetic fruit juices, and vegetable beverages. These ingredients may be used alone or in combination. Furthermore, the health functional food composition may be in any of the following forms: meat, sausage, bread, chocolate, candy, snacks, pastries, pizza, instant noodles, chewing gum, ice cream, soup, beverages, tea, functional water, drinks, alcohol, and vitamin complexes.

[0061] Furthermore, the aforementioned health functional foods may further contain food additives, and unless otherwise specified, their suitability as "food additives" shall be determined in accordance with the general rules and general testing methods of the Korean Food Additives Code approved by the Ministry of Food and Drug Safety of Korea, based on the relevant standards and guidelines for the corresponding items.

[0062] The items listed in the Korean Food Additives Code may include, for example: chemically synthesized compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon pigments, licorice extract, crystalline cellulose, sorghum pigments, and guar gum; and mixed preparations such as L-glutamate preparations, alkali agents for noodles, preservatives, and tar coloring agents.

[0063] In this article, the active ingredient added to the food during the production of the health functional food can be adjusted as needed, and preferably, it can be added in an amount of 1 to 90 parts by weight per 100 parts by weight of the food.

[0064] Example

[0065] The present disclosure will be described in more detail below by way of examples to aid in understanding the present disclosure. However, the following examples are intended to illustrate the present disclosure only, and the scope of the present disclosure is not limited to the following examples. Examples of the present disclosure are provided to explain the present disclosure more fully to those skilled in the art.

[0066] [Synthetic Examples] Synthesis of Arylpiperidine Derivatives

[0067] (1) Synthesis method A

[0068] Piperidine derivative (2.92 mmol), fluorobenzene (3.21 mmol), potassium carbonate (5.84 mmol), pyridine (3.50 mmol), and DMSO (5 mL) were sequentially added to a 50 mL round-bottom flask. The reaction flask was placed in an oil bath preheated to 90 °C and stirred overnight. The resulting mixture was then cooled to room temperature, diluted with ethyl acetate (EtOAc), and washed thoroughly with water. The solvent was removed under vacuum, and the residue was purified by column chromatography to obtain the desired compound.

[0069] (2) Synthesis method B

[0070] Lithium hydroxide (6.52 mmol) dissolved in water (6 mL) was added to a mixed solution of tetrahydrofuran (11 mL) and methanol (11 mL) containing the compound (2.17 mmol) produced by synthesis method A described above. The mixture was then stirred overnight at room temperature and subsequently evaporated to remove methanol. The reaction mixture was acidified with 1 M hydrochloric acid and then extracted with ethyl acetate (EtOAc). The combined organic phases were washed with brine, dried over sodium sulfate, filtered, and evaporated to give the desired compound.

[0071] (3) Synthesis method C

[0072] EDC (0.413 mmol) and DMAP (0.636 mmol) were added to 1 mL of a DMF solution containing 0.318 mmol of 2-(1,4-dioxo-3,4-dihydrophthalazine-2(1H)-yl)acetic acid. The mixture was then stirred at room temperature for 30 minutes, an amine (0.382 mmol) was added, and the mixture was stirred overnight at room temperature. The reaction mixture was diluted with water and ethyl acetate (EtOAc) and extracted twice with ethyl acetate (EtOAc). The combined organic layers were washed with brine, washed twice with water, dried over sodium sulfate, filtered, and evaporated. The mixture was purified by column chromatography to give the desired compound.

[0073] (4) Ethyl 2-(1-(2-cyanophenyl)piperidin-4-yl)

[0074]

[0075] 2-(1-(2-cyanophenyl)piperidin-4-yl)ethyl acetate was synthesized according to the above-mentioned synthetic method A.

[0076] 1H NMR (300 MHz, CDCl3) δ 7.54 (dd, J = 7.7, 1.7 Hz, 1H), 7.45 (td, J= 7.7, 1.7 Hz, 1H), 7.04 - 6.90 (m, 2H), 4.14 (q, J = 7.1 Hz, 2H), 3.57 (t, J= 11.5 Hz, 2H), 2.81 (t, J = 12.0 Hz, 2H), 2.30 (d, J = 6.9 Hz, 2H), 2.02 -1.80 (m, 3H), 1.60 - 1.45 (m, 2H), 1.27 (t, J = 7.1 Hz, 3H).

[0077] (5) 2-(1-(2-cyanophenyl)piperidin-4-yl)acetic acid

[0078]

[0079] 2-(1-(2-cyanophenyl)piperidin-4-yl)acetic acid was synthesized according to the above-mentioned synthetic method B.

[0080] 1 H NMR (300 MHz, CDCl3) δ 7.54 (dd, J = 7.6, 1.6 Hz, 1H), 7.46 (td, J= 7.6, 1.6 Hz, 1H), 7.04 - 6.93 (m, 2H), 3.58 (t, J = 11.8 Hz, 2H), 2.82 (t,J = 12.0 Hz, 2H), 2.37 (d, J = 6.8 Hz, 2H), 2.06 - 1.84 (m, 3H), 1.67 - 1.48 (m, 2H).

[0081] (6) Ethyl 3-(1-(2-cyanophenyl)piperidin-4-yl)propionate

[0082]

[0083] Ethyl 3-(1-(2-cyanophenyl)piperidin-4-yl)propionate was synthesized according to the above-mentioned synthesis method A.

[0084] 1H NMR (300 MHz, CDCl3) δ 7.53 (dd, J = 7.6, 1.6 Hz, 1H), 7.45 (td, J= 8.6, 1.6 Hz, 1H), 7.03 - 6.91 (m, 2H), 4.14 (q, J = 7.1 Hz, 2H), 3.58 (d, J= 12.8 Hz, 2H), 2.77 (t, J = 10.8 Hz, 2H), 2.36 (t, J = 7.7 Hz, 2H), 1.82 (d,J = 9.6 Hz, 2H), 1.70 - 1.61 (m, 3H), 1.45 (d, J = 6.5 Hz, 2H), 1.26 (t, J =7.1 Hz, 3H).

[0085] (8) 3-(1-(2-cyanophenyl)piperidin-4-yl)propionic acid

[0086]

[0087] 3-(1-(2-cyanophenyl)piperidin-4-yl)propionic acid was synthesized according to the above-mentioned synthesis method B.

[0088] 1 H NMR (300 MHz, CDCl3) δ 7.54 (dd, J = 7.6, 1.7 Hz, 1H), 7.45 (td, J= 8.7, 1.7 Hz, 1H), 7.03 - 6.91 (m, 2H), 3.65 - 3.51 (m, 2H), 2.77 (t, J =11.2 Hz, 2H), 2.43 (t, J = 7.6 Hz, 2H), 1.91 - 1.77 (m, 2H), 1.68 (q, J = 7.6Hz, 2H), 1.58 - 1.38 (m, 3H).

[0089] (9) 4-(1-(2-cyanophenyl)piperidin-4-yl)butyric acid

[0090]

[0091] 4-(1-(2-cyanophenyl)piperidin-4-yl)butyric acid was synthesized according to the above-mentioned synthesis method B.

[0092] 1H NMR (300 MHz, CDCl3) δ 7.54 (dd, J = 7.7, 1.6 Hz, 1H), 7.50 - 7.40(m, 1H), 7.08 - 6.90 (m, 2H), 3.66 - 3.51 (m, 2H), 2.88 - 2.72 (m, 2H), 2.38(t, J = 7.4 Hz, 2H), 1.83 (d, J = 11.2 Hz, 2H), 1.77 - 1.63 (m, 2H), 1.58 -1.30 (m, 5H).

[0093] (10) TI-61815 (N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-2-(1-(2-cyanophenyl)piperidin-4-yl)acetamide)

[0094]

[0095] TI-61815 was synthesized according to the above synthesis method C (yield 50%).

[0096] 1 H NMR (300 MHz, DMSO-d6) δ 10.96 (s, 1H), 7.98 (d, J = 7.0 Hz, 1H), 7.78 - 7.66 (m, 2H), 7.62 - 7.54 (m, 1H), 7.43 - 7.35 (m, 1H), 7.17 (d, J =8.3 Hz, 1H), 7.10 - 6.96 (m, 2H), 3.57 - 3.46 (m, 2H), 2.90 - 2.76 (m, 2H),2.51 (d, J = 5.4 Hz, 2H), 1.90 (d, J = 13.5 Hz, 3H), 1.58 - 1.39 (m, 2H).

[0097] (11) 2-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-b]pyridazin-3-yl)acetamide (hereinafter referred to as TI-61816)

[0098]

[0099] TI-61816 was synthesized according to the above synthesis method C (yield 60%).

[0100] 1H NMR (300 MHz, CDCl3) δ 8.48 - 8.38 (m, 1H), 8.35 (s, 1H), 8.25 -8.02 (m, 2H), 7.54 (dd, J = 7.6, 1.7 Hz, 1H), 7.50 - 7.41 (m, 1H), 7.09 (dd,J = 9.2, 4.5 Hz, 1H), 7.04 - 6.93 (m, 2H), 3.65 - 3.55 (m, 2H), 2.85 (t, J =12.0 Hz, 2H), 2.55 (d, J = 7.1 Hz, 2H), 2.24 - 2.10 (m, 1H), 1.96 (d, J =13.0, 3.5 Hz, 2H), 1.72 - 1.54 (m, 2H).

[0101] (12) 2-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-a]pyridin-3-yl)acetamide (hereinafter referred to as TI-61817)

[0102]

[0103] TI-61817 was synthesized according to the above synthesis method C (yield 84%).

[0104] 1 H NMR (300 MHz, DMSO-d6) δ 10.10 (s, 1H), 8.05 (d, J = 6.9 Hz, 1H), 7.69 (dd, J = 7.7, 1.6 Hz, 1H), 7.64-7.51 (m, 2H), 7.48 (s, 1H), 7.29-7.13(m, 2H), 7.07 (t, J = 7.6, 1.0 Hz, 1H), 6.96 (t, J = 6.9 Hz, 1H), 3.51 (d, J= 11.7 Hz, 2H), 2.83 (t, J = 12.9, 10.6 Hz, 2H), 2.45 (d, J = 6.9 Hz, 2H), 2.07-1.82 (m, 3H), 1.57-1.39 (m, 2H).

[0105] (13) 2-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-a]pyridin-5-yl)acetamide (hereinafter referred to as TI-61818)

[0106]

[0107] TI-61818 was synthesized according to the above synthesis method C (yield 77%).

[0108] 1 H NMR (300 MHz, DMSO-d6) δ 10.34 (s, 1H), 7.85 (d, J = 1.3 Hz, 1H), 7.65 (dd, J = 7.7, 1.6 Hz, 1H), 7.61 - 7.49 (m, 2H), 7.41 (d, J = 9.0 Hz,1H), 7.24 (dd, J = 9.0, 7.2 Hz, 1H), 7.13 (d, J = 8.3 Hz, 1H), 7.03 (t, J =7.5 Hz, 2H), 3.47 (d, J = 11.7 Hz, 2H), 2.88 - 2.71 (m, 2H), 2.47 (d, J =3.7, 1.8 Hz, 2H), 2.04 - 1.91 (m, 1H), 1.89 - 1.78 (m, 2H), 1.55 - 1.36 (m, 2H).

[0109] (14) TI-61910 (N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(2-cyanophenyl)piperidin-4-yl)propionamide)

[0110]

[0111] TI-61910 was synthesized according to the above synthesis method C (yield 50%).

[0112] 1H NMR (300 MHz, DMSO-d6) δ 10.92 (s, 1H), 8.00 (d, J = 7.0 Hz, 1H), 7.82 - 7.64 (m, 2H), 7.57 (td, J = 7.9, 1.7 Hz, 1H), 7.44 - 7.31 (m, 1H),7.16 (d, J = 8.3 Hz, 1H), 7.10 - 6.89 (m, 2H), 3.51 (d, J = 11.9 Hz, 2H),2.78 (t, J = 11.5 Hz, 2H), 2.56 (t, J = 7.8 Hz, 2H), 1.85 (d, J = 12.0 Hz,2H), 1.68 (q, J = 7.3 Hz, 2H), 1.57 - 1.28 (m, 3H).

[0113] (15) N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-4-(1-(2-cyanophenyl)piperidin-4-yl)butyramide (hereinafter referred to as TI-62116)

[0114]

[0115] TI-62116 was synthesized according to the above synthesis method C (yield 55%).

[0116] 1 H NMR (300 MHz, DMSO-d6) δ 10.89 (s, 1H), 7.99 (d, J = 6.9 Hz, 1H), 7.79 - 7.63 (m, 2H), 7.56 (d, J = 8.1 Hz, 1H), 7.38 (t, J = 8.1 Hz, 1H), 7.15(d, J = 8.4 Hz, 1H), 7.09 - 6.94 (m, 2H), 3.50 (d, J = 11.6 Hz, 2H), 2.77 (t,J = 11.6 Hz, 2H), 1.91 - 1.62 (m, 4H), 1.37 (q, J = 17.1 Hz, 5H).

[0117] (16) 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-b]pyridazin-3-yl)propionamide (hereinafter referred to as TI-62121)

[0118]

[0119] TI-62121 was synthesized according to the above synthesis method C (yield 82%).

[0120] 1 H NMR (300 MHz, DMSO-d6) δ 10.47 (s, 1H), 8.55 (dd, J = 4.4, 1.5 Hz,1H), 8.09 (dd, J = 9.2, 1.6 Hz, 1H), 7.86 (s, 1H), 7.67 (dd, J = 7.7, 1.6 Hz,1H), 7.61 - 7.53 (m, 1H), 7.22 - 7.12 (m, 2H), 7.05 (t, J = 7.5 Hz, 1H), 3.51(d, J = 11.9 Hz, 2H), 2.77 (t, J = 10.9 Hz, 2H), 2.56 (t, J = 7.6 Hz, 2H),1.90 - 1.78 (m, 2H), 1.69 - 1.59 (m, 2H), 1.54 - 1.27 (m, 4H).

[0121] (17) 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-a]pyridin-3-yl)propionamide (hereinafter referred to as TI-62122)

[0122]

[0123] TI-62122 was synthesized according to the above synthesis method C (yield 50%).

[0124] 1 H NMR (300 MHz, DMSO-d6) δ 10.04 (s, 1H), 8.05 (d, J = 6.8 Hz, 1H), 7.71 - 7.64 (m, 1H), 7.57 (dd, J = 13.0, 8.4 Hz, 2H), 7.46 (s, 1H), 7.28 -7.13 (m, 2H), 7.05 (t, J = 7.5 Hz, 1H), 6.94 (t, J = 6.8 Hz, 1H), 3.52 (d, J= 11.8 Hz, 2H), 3.17 (d, J = 5.3 Hz, 2H), 2.79 (t, J = 11.5 Hz, 2H), 1.86 (d,J = 11.9 Hz, 2H), 1.74 - 1.62 (m, 2H), 1.56 - 1.31 (m, 3H).

[0125] (18) 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-a]pyridin-5-yl)propionamide (hereinafter referred to as TI-62123)

[0126]

[0127] TI-62123 was synthesized according to the above synthesis method C (yield 50%).

[0128] 1 H NMR (300 MHz, DMSO-d6) δ 10.34 (s, 1H), 7.88 (s, 1H), 7.68 (dd, J =7.7, 1.6 Hz, 1H), 7.64 - 7.51 (m, 2H), 7.44 (d, J = 9.0 Hz, 1H), 7.27 (dd, J= 9.0, 7.2 Hz, 1H), 7.15 (d, J = 8.3 Hz, 1H), 7.12 - 7.00 (m, 2H), 3.52 (d, J= 11.3 Hz, 2H), 2.78 (t, J = 12.0 Hz, 2H), 2.63 - 2.54 (m, 2H), 1.85 (d, J =11.9 Hz, 2H), 1.68 (q, J = 7.1 Hz, 2H), 1.57 - 1.29 (m, 3H).

[0129] (19) 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(1-methyl-1H-indole-6-yl)propionamide (hereinafter referred to as TI-62125)

[0130]

[0131] TI-62125 was synthesized according to the above synthesis method C (yield 61%).

[0132] 1H NMR (300 MHz, DMSO-d6) δ 9.86 (s, 1H), 7.94 (s, 1H), 7.73 - 7.64(m, 1H), 7.55 (d, J = 8.7 Hz, 1H), 7.43 (d, J = 8.4 Hz, 1H), 7.23 (d, J = 3.1Hz, 1H), 7.14 (d, J = 8.4 Hz, 1H), 7.06 (d, J = 7.9 Hz, 2H), 6.33 (d, J = 3.1Hz, 1H), 3.72 (s, 3H), 3.50 (d, J = 11.7 Hz, 2H), 2.76 (t, J = 11.4 Hz, 2H),2.39 (t, J = 7.6 Hz, 2H), 1.84 (d, J = 11.7 Hz, 2H), 1.72 - 1.59 (m, 2H), 1.51 - 1.28 (m, 3H).

[0133] (20) 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(1H-indol-4-yl)propionamide (hereinafter referred to as TI-62126)

[0134]

[0135] TI-62126 was synthesized according to the above synthesis method C (yield 66%).

[0136] 1 H NMR (300 MHz, DMSO-d6) δ 11.09 (s, 1H), 9.52 (s, 1H), 7.67 (d, J =7.7 Hz, 1H), 7.64 - 7.50 (m, 2H), 7.28 (t, J = 2.8 Hz, 1H), 7.14 (dd, J =8.3, 3.4 Hz, 2H), 7.09 - 6.94 (m, 2H), 6.70 (s, 1H), 3.51 (d, J = 11.7 Hz, 2H), 2.76 (t, J = 11.4 Hz, 2H), 1.86 (d, J = 11.7 Hz, 2H), 1.66 (q, J = 7.2Hz, 2H), 1.55 - 1.29 (m, 3H).

[0137] 13C NMR (101 MHz, DMSO-d6) δ 171.39, 156.05, 136.68, 134.21, 134.13,130.75, 124.02, 121.51, 121.08, 120.26, 119.10, 118.38, 110.51, 107.36,104.79, 99.17, 51.92, 34.58, 33.52, 31.95.

[0138] (21) 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(1H-indol-7-yl)propionamide (hereinafter referred to as TI-62127)

[0139]

[0140] TI-62127 was synthesized according to the above synthesis method C (yield 83%).

[0141] 1 H NMR (300 MHz, DMSO-d6) δ 10.67 (s, 1H), 9.66 (s, 1H), 7.73 - 7.64(m, 1H), 7.61 - 7.53 (m, 1H), 7.43 - 7.28 (m, 3H), 7.15 (d, J = 8.4 Hz, 1H),7.05 (t, J = 7.5 Hz, 1H), 6.93 (t, J = 7.7 Hz, 1H), 6.43 (t, J = 2.5 Hz, 1H),3.52 (d, J = 11.8 Hz, 2H), 2.78 (t, J = 11.6 Hz, 2H), 1.87 (d, J = 12.0 Hz, 2H), 1.75 - 1.63 (m, 2H), 1.54 - 1.29 (m, 3H).

[0142] (22) N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(2-nitrophenyl)piperidin-4-yl)propionamide (hereinafter referred to as TI-62188)

[0143]

[0144] TI-62188 was synthesized according to the above synthesis method C (yield 61%).

[0145] 1H NMR (300 MHz, DMSO-d6) δ 10.90 (s, 1H), 7.99 (d, J = 7.0 Hz, 1H), 7.85 - 7.70 (m, 2H), 7.60 - 7.51 (m, 1H), 7.43 - 7.26 (m, 2H), 7.13 - 6.95(m, 2H), 3.26 - 3.12 (m, 2H), 2.87 - 2.73 (m, 2H), 2.55 (d, J = 8.0 Hz, 2H), 1.85 - 1.60 (m, 4H), 1.54 - 1.22 (m, 3H).

[0146] (23) N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(3-nitrophenyl)piperidin-4-yl)propionamide (hereinafter referred to as TI-62189)

[0147]

[0148] TI-62189 was synthesized according to the above synthesis method C (yield 45%).

[0149] 1 H NMR (300 MHz, DMSO-d6) δ 10.89 (s, 1H), 7.99 (d, J = 7.0 Hz, 1H), 7.74 (d, J = 9.3 Hz, 1H), 7.64 (d, J = 2.3 Hz, 1H), 7.53 (d, J = 7.5 Hz, 1H),7.49 - 7.33 (m, 3H), 6.98 (t, J = 6.7 Hz, 1H), 3.86 (d, J = 12.4 Hz, 2H), 2.80 (t, J = 12.1 Hz, 2H), 2.56 (d, J = 8.1 Hz, 2H), 1.82 (d, J = 12.6 Hz,2H), 1.73 - 1.45 (m, 3H), 1.35 - 1.18 (m, 2H).

[0150] (24) N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(4-nitrophenyl)piperidin-4-yl)propionamide (hereinafter referred to as TI-62190)

[0151]

[0152] TI-62190 was synthesized according to the above synthesis method C (yield 89%).

[0153] 1 H NMR (300 MHz, DMSO-d6) δ 8.08 - 7.93 (m, 3H), 7.70 (d, J = 9.3 Hz,1H), 7.35 (dd, J = 9.3, 6.5 Hz, 1H), 7.07 - 6.90 (m, 3H), 4.07 (d, J = 13.2Hz, 2H), 2.98 (t, J = 12.5 Hz, 2H), 2.57 - 2.52 (m, 1H), 1.80 (d, J = 12.9Hz, 2H), 1.69 - 1.53 (m, 3H), 1.28 - 1.09 (m, 2H).

[0154] 13 C NMR (101 MHz, DMSO-d6) δ 173.31, 154.47, 148.39, 141.42, 136.06,127.39, 125.91, 124.06, 115.42, 112.90, 112.39, 46.83, 34.88, 32.57, 31.25, 31.04.

[0155] (25) N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(4-cyanophenyl)piperidin-4-yl)propionamide (hereinafter referred to as TI-62194)

[0156]

[0157] TI-62194 was synthesized according to the above synthesis method C (yield 45%).

[0158] 1H NMR (300 MHz, DMSO-d6) δ 10.90 (s, 1H), 7.98 (d, J = 7.0 Hz, 1H), 7.78 - 7.70 (m, 1H), 7.60 - 7.50 (m, 2H), 7.43 - 7.34 (m, 1H), 7.07 - 6.93(m, 3H), 3.95 (d, J = 12.9 Hz, 2H), 2.85 (t, J = 12.4 Hz, 2H), 2.54 (d, J =7.7 Hz, 2H), 1.78 (d, J = 12.9 Hz, 2H), 1.62 (d, J = 7.1 Hz, 3H), 1.27 - 1.12(m, 2H).

[0159] (26) N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(2,4-dinitrophenyl)piperidin-4-yl)propionamide (hereinafter referred to as TI-62195)

[0160]

[0161] TI-62195 was synthesized according to the above synthesis method C (yield 51%).

[0162] 1 H NMR (300 MHz, DMSO-d6) δ 10.88 (s, 1H), 8.59 (d, J = 2.7 Hz, 1H), 8.24 (dd, J = 9.4, 2.7 Hz, 1H), 8.00 (d, J = 6.9 Hz, 1H), 7.73 (d, J = 9.4Hz, 1H), 7.46 - 7.32 (m, 2H), 6.98 (t, J = 6.7 Hz, 1H), 3.47 (d, J = 13.1 Hz, 2H), 3.17 - 3.04 (m, 2H), 2.55 (d, J = 6.8 Hz, 2H), 1.82 (d, J = 13.1 Hz,2H), 1.72 - 1.54 (m, 3H), 1.40 - 1.22 (m, 2H).

[0163] (27) 2-(1,4-dioxa-8-azaspiro[4.5]dec-8-yl)benzonitrile

[0164]

[0165] 2-(1,4-dioxa-8-azaspiro[4.5]dec-8-yl)benzonitrile was synthesized according to the above-mentioned synthesis method A.

[0166] 1 H NMR (300 MHz, CDCl3) δ 7.55 (dd, J = 7.7, 1.6 Hz, 1H), 7.50 - 7.42(m, 1H), 7.07 - 6.95 (m, 2H), 4.00 (s, 4H), 3.35 - 3.26 (m, 4H), 1.97 - 1.89(m, 4H).

[0167] (28) 2-(4-oxopiperidin-1-yl)benzonitrile

[0168]

[0169] The 2-(1,4-dioxa-8-azaspiro[4.5]dec-8-yl)benzonitrile (248 mg, 1.02 mmol) was dissolved in ethanol (3 mL), and then refluxed with hydrochloric acid (1 M, 1 mL) for 5 hours. Subsequently, an aqueous sodium bicarbonate solution was added to the reaction solution, and the organic layer was extracted three times with ethyl acetate (EtOAc). The combined organic layers were dried over sodium sulfate, filtered, and evaporated. The mixture was purified by column chromatography to give the 2-(4-oxopiperidin-1-yl)benzonitrile compound (164 mg, 81%).

[0170] 1 H NMR (300 MHz, CDCl3) δ 7.61-7.55(m,1H),7.50 (td, J = 7.9, 1.7 Hz,1H), 7.06 (dd, J = 8.1, 5.9 Hz, 2H), 3.49 (t, J = 6.0 Hz, 4H), 2.66 (t, J =6.0 Hz, 4H).

[0171] (29) (1-(2-cyanophenyl)piperidin-4-yl)glycine ethyl ester

[0172]

[0173] Glycine ethyl ester hydrochloride (139 mg, 0.999 mmol), TEA (0.138 mL, 0.999 mmol), and sodium triacetoxyborohydride (529 mg, 2.50 mmol) were added to the 2-(4-oxopiperidin-1-yl)benzonitrile compound (100 mg, 0.499 mmol) dissolved in dichloromethane (CH2Cl2, 3.3 mL). The reaction mixture was stirred at room temperature for 24 hours. The reaction mixture was diluted with dichloromethane (CH2Cl2), washed with sodium bicarbonate solution and brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to give (1-(2-cyanophenyl)piperidin-4-yl)glycine ethyl ester compound (119 mg, 83%).

[0174] 1 H NMR (300 MHz, CDCl3) δ 7.53 (dd, J = 7.6, 1.6 Hz, 1H), 7.48 - 7.39(m, 1H), 7.01 - 6.91 (m, 2H), 4.20 (q, J = 7.2 Hz, 2H), 3.62 - 3.51 (m, 2H),3.47 (s, 2H), 2.94 - 2.81 (m, 2H), 2.75 - 2.62 (m, 1H), 2.08 - 1.94 (m, 3H),1.73 - 1.59 (m, 2H), 1.28 (t, J = 7.2 Hz, 3H).

[0175] (30) N-(tert-Butoxycarbonyl)-N-(1-(2-cyanophenyl)piperidin-4-yl)glycine ethyl ester

[0176]

[0177] A solution mixture of the (1-(2-cyanophenyl)piperidin-4-yl)glycine ethyl ester compound (300 mg, 1.04 mmol), di-tert-butyl dicarbonate (273 mg, 1.25 mmol), and potassium carbonate (433 mg, 3.13 mmol) in MeCN (5 mL) was stirred for 6 hours. The reaction mixture was then filtered and concentrated. The residue was purified by column chromatography to give N-(tert-butyloxycarbonyl)-N-(1-(2-cyanophenyl)piperidin-4-yl)glycine ethyl ester compound (377 mg, 93% yield).

[0178] 1H NMR (300 MHz, CDCl3) δ 7.57 (d, J = 7.9 Hz, 1H), 7.49 (t, J = 7.9Hz, 1H), 7.21 - 6.98 (m, 2H), 4.40 - 4.24 (m, 1H), 4.19 (q, J = 7.1 Hz, 2H), 3.89 (d, J = 32.7 Hz, 2H), 3.61 (d, J = 11.7 Hz, 2H), 2.97 (d, J = 13.9 Hz, 2H), 1.87 (d, J = 20.5 Hz, 4H), 1.47 (s, 9H), 1.29 (t, J = 7.1 Hz, 3H).

[0179] (31) N-(tert-Butoxycarbonyl)-N-(1-(2-cyanophenyl)piperidin-4-yl)glycine

[0180]

[0181] Lithium hydroxide (120 mg, 2.87 mmol) dissolved in water (2 mL) was added to a solution of N-(tert-butoxycarbonyl)-N-(1-(2-cyanophenyl)piperidin-4-yl)glycine ethyl ester (370 mg, 0.955 mmol) in tetrahydrofuran (3 mL) and methanol (3 mL) as solvents. The mixture was stirred overnight at room temperature and then evaporated to remove methanol. The reaction mixture was acidified with 1 M hydrochloric acid and then extracted with ethyl acetate (EtOAc). The combined organic phases were washed with brine, dried over sodium sulfate, filtered, and evaporated to give N-(tert-butoxycarbonyl)-N-(1-(2-cyanophenyl)piperidin-4-yl)glycine compound (336 mg, 98% yield).

[0182] 1 H NMR (300 MHz, CDCl3) δ 7.60 - 7.42 (m, 2H), 7.03 (q, J = 8.3 Hz,2H), 4.43 - 4.14 (m, 1H), 3.95 (s, 2H), 3.61 (d, J = 11.9 Hz, 2H), 2.92 (t, J= 11.9 Hz, 2H), 2.00 - 1.70 (m, 4H), 1.47 (s, 9H).

[0183] (32) (2-([1,2,4]triazolo[4,3-a]pyridin-3-ylamino)-2-oxoethyl)(1-(2-cyanophenyl)piperidin-4-yl)carbamate tert-butyl ester (hereinafter referred to as TI-62197)

[0184]

[0185] TI-62197 was synthesized according to the above synthesis method C (yield 51%).

[0186] 1 H NMR (300 MHz, CDCl3) δ 8.15 (d, J = 7.0 Hz, 1H), 7.81 (d, J = 9.3Hz, 1H), 7.56 - 7.40 (m, 3H), 6.99 (t, J = 7.7 Hz, 3H), 4.27 (s, 3H), 3.63 (d, J = 11.6 Hz, 2H), 2.97 - 2.81 (m, 2H), 1.99 (d, J = 10.1 Hz, 4H), 1.49 (s, 9H).

[0187] (33) N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-2-((1-(2-cyanophenyl)piperidin-4-yl)amino)acetamide trifluoroacetate (hereinafter referred to as TI-62198)

[0188]

[0189] At 0°C, a 20% trifluoroacetic acid solution dissolved in dichloromethane (CH2Cl2) was added to a solution of TI-62197 (60 mg, 0.298 mmol) in anhydrous dichloromethane (1 mL; CH2Cl2) as a solvent. The reaction mixture was stirred at room temperature for 1 hour, diluted with dichloromethane (CH2Cl2), and evaporated several times with diethyl ether to give compound TI-62198 (95% yield).

[0190] 1H NMR (300 MHz, DMSO-d6) δ 11.63 (s, 1H), 9.32 (s, 2H), 8.20 (d, J =7.0 Hz, 1H), 7.83 - 7.69 (m, 2H), 7.61 (t, J = 7.9 Hz, 1H), 7.49 - 1.90 - 1.74 (m, 2H).

[0191] (34) N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-2-((1-(2-cyanophenyl)piperidin-4-yl)(methyl)amino)acetamide (hereinafter referred to as TI-62199)

[0192]

[0193] To a solution of TI-62198 (40 mg, 0.0820 mmol) stirred in MeCN (1 mL) as a solvent, 37% formaldehyde solution (800 μL, 0.286 mmol) and sodium triacetoxyborohydride (69.3 mg, 0.327 mmol) were added, and the reaction mixture was then stirred at room temperature for 3 hours. Water was added to the reaction mixture, and the mixture was extracted three times with dichloromethane (CH2Cl2). The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give the crude product. The residue was purified by column chromatography to give compound TI-62199 (13 mg, 41% yield).

[0194] 1H NMR (300 MHz, CDCl3) δ 8.14 (s, 1H), 7.67 (dd, J = 9.3, 1.3 Hz,1H), 7.56 (dd, J = 7.9, 1.7 Hz, 1H), 7.48 (td, J = 7.9, 1.7 Hz, 1H), 7.32 -7.26 (m, 1H), 7.06 - 6.97 (m, 2H), 6.90 - 6.82 (m, 1H), 3.75 - 3.52 (m, 4H), 3.02 - 2.77 (m, 3H), 2.66 (t, J = 2.7 Hz, 3H), 2.15 - 2.04 (m, 2H), 1.99 -1.84 (m, 2H).

[0195] [Preparation Example 1] Cell Preparation

[0196] All cell lines were purchased from the American Type Culture Collection (ATCC) and cultured at 37°C in the presence of 5% CO2 in RPMI 1640 (Roswell Park Memorial Institute 1640) medium supplemented with 10% fetal bovine serum (FBS), 100 μg / mL streptomycin, and 100 units / mL penicillin.

[0197] [Preparation Example 2] Positive Control Group

[0198] In all end-anchored polymerase experiments, 2-(4-(trifluoromethylphenyl)-3,5,7,8-tetrahydro-thiarano[4,3-d]pyrimidin-4-one (XAV939) was used as a positive control.

[0199] In the experiment on the anti-enzyme ability of PARP-1, olaparib was used as a positive control group.

[0200] [Example 1] Determination of anti-enzyme ability against end-anchored polymerase (TNKS)

[0201] To determine whether the arylpiperidine derivative could inhibit the enzymatic activity of the end-anchor polymerase, the end-anchor polymerase-1 colorimetric activity assay kit (TNKS1 Histone Ribosylation Colorimetric Assay Kit, Cat# 80582, BPS Bioscience, USA) was used.

[0202] In addition, to determine whether the arylpiperidine derivatives inhibit the enzymatic activity of poly-ADP-ribose polymerase (PARP), the compounds were treated with a PARP-1 colorimetric activity assay kit (Cat# 80580, BPS Bioscience) at 1 μM each, according to the manufacturer's instructions, for measurement and identification.

[0203] The results of TNKS-1 or PARP-1 inhibition are shown in Table 1 below.

[0204] When XAV939, used as a positive control group for end-anchor polymerase, was treated with 1 μM, it inhibited the enzyme activities of both TNKS-1 and TNKS-2 by approximately 100%; while when olaparib was treated with 1 μM, it inhibited the PARP enzyme activity by 100%.

[0205] Table 1

[0206] compound TNKS-1 inhibitory activity (10 μM) TI-61815 100 TI-61816 5±1.2 TI-61910 100.0 TI-62116 99±0.1 TI-62121 17±1.5 TI-62122 10±0.9 TI-62126 1±1.7 TI-62187 100.0 TI-62188 100.0 TI-62189 100.0 TI-62190 96±0.2 TI-62194 100.0 TI-62195 100.0 TI-62197 3±0.9 TI-62198 72±0.3 TI-62199 98±0.1

[0207] Table 2

[0208] <![CDATA[TNKS-1 IC 50 (μM)]]> <![CDATA[TNKS-2 IC 50 (μM)]]> <![CDATA[PARP-1 IC 50 (μM)]]> <![CDATA[PARP-2 IC 50 (μM)]]> TI-61815 0.064 0.049 7.54 6.062 TI-61910 0.018 0.004 >10 >10

[0209] As shown in Table 1 above, the four types of arylpiperidine derivatives (TI-61815, TI-61910, TI-62188, and TI-62195) inhibited the enzyme activity of TNKS-1 by 95% or higher, and TI-61815 and TI-61910 showed almost no reduction in the inhibition of PARP-1 enzyme activity. Therefore, the inhibitory concentrations (IC50) of the two types of arylpiperidine derivatives against TNKS-1 and TNKS-2 were determined. 50 This concentration induces a maximum inhibition of 50%.

[0210] The results are shown in Table 2, which determined the IC values ​​of the two types of derivatives. 50 The TI-61910 has the lowest IC value for both TNKS-1 and TNKS-2. 50 The values ​​were high, but neither PARP-1 nor PARP-2 were inhibited at concentrations below 10 μM.

[0211] [Example 2] Identification of inhibition of intracellular β-catenin transcriptional activity

[0212] To determine whether the arylpiperidine derivatives inhibited the transcriptional activity of β-catenin, the cells were treated with compounds TI-61815 and TI-61910 at different concentrations, and the results were detected in HEK cells using a TCF / luciferase detection system.

[0213] The results are shown in Table 3, which identified TI-61815 and TI-61910 as having a 50% inhibitory effect (IC50) on the transcriptional activity of the β-catenin. 50 The TI-61910 showed the lowest possible inhibition of β-catenin transcriptional activity.

[0214] Table 3

[0215] <![CDATA[IC of β-catenin in HEK cells 50 (μM)]]> TI-61815 0.83 TI-61910 0.49

[0216] [Example 3] Identification of the repression of β-catenin expression and the stabilization of AXIN2 expression

[0217] To determine whether the arylpiperidine derivatives inhibited β-catenin protein expression, colon cancer cell lines COLO320DM and SW403, treated with 10 μM of the two compounds (TI-61815 and TI-61910), were cultured at 37°C in the presence of 5% CO2 for 24 hours, and then the cells were collected. After protein extraction, the proteins were quantified using Western blotting (Bio-Rad, USA).

[0218] The results are as follows Figure 2 As shown, TI-61815 and TI-61910 inhibited the protein level of the activated β-catenin and determined the stability of the AXIN2 protein level.

[0219] [Example 4] Identification of the repression of β-catenin subgene expression

[0220] To determine whether the arylpiperidine derivatives inhibited the expression of the β-catenin subgene, the colon cancer cell line COLO320DM, treated with 10 μM of both compounds (TI-61815 and TI-61910), was cultured at 37°C in the presence of 5% CO2 for 24 hours, and then the cells were collected. RNA was extracted, and cDNA was synthesized for quantification of gene expression by real-time PCR.

[0221] The results are as follows Figure 3 As shown, TI-61815 and TI-61910 inhibited the expression levels of β-catenin subgenes (AXIN2, CCND1, cMYC, FGF20).

[0222] [Example 5] Identifying cell viability

[0223] Cell viability was assessed by treating the colon cancer cell line COLO320DM with TI-61815 and TI-61910.

[0224] After treatment with TI-61815 and TI-61910 at concentrations ranging from 2.5 μM to 40 μM, the cells were cultured at 37 °C in the presence of 5% CO2 for 10 days. Cell viability was assessed by counting the number of colonies using 1.5% methylene blue staining.

[0225] The results are as follows Figure 4 As shown, when treated with TI-61815 and TI-61910, the cell viability of the colon cancer cell line decreased in a concentration-dependent manner, and when treated with 40 μM TI-61815, cell viability was inhibited by up to 61.4%, and when treated with 40 μM TI-61910, cell viability was inhibited by up to 60.8%.

[0226] [Example 6] Identification of the anticancer effect of 5-FU combined treatment

[0227] The colon cancer cell line COLO320DM was treated with 5 μM or 10 μM TI-61815 in combination with 0.5 μM 5-FU, or with 2.5 μM, 5 μM, or 10 μM TI-61910 in combination with 0.5 μM of the anticancer drug 5-fluorouracil (5-FU), and then cultured at 37°C in the presence of 5% CO2 for 10 days. Cell viability was then assessed by counting the number of colonies using 1.5% methylene blue staining.

[0228] As a result, according to Figure 5 Compared with TI-61815 alone (5 μM or 10 μM), cell viability was inhibited by 12.4% and 27.8% when treated with 5-FU, respectively; and compared with TI-61910 alone (2.5 μM, 5 μM, or 10 μM), cell viability was inhibited by 26%, 28.3%, and 35.5% when treated with 5-FU, respectively.

[0229] The foregoing description is for illustrative purposes only, and those skilled in the art will understand that this disclosure can be readily modified into other specific forms without altering its technical concept or essential characteristics. Therefore, it should be understood that the embodiments shown above are exemplary and not restrictive in all respects.

[0230] The scope of this disclosure is defined by the appended claims, and all changes or modifications arising from the meaning and scope of the claims and their equivalents shall be construed as being included within the scope of this disclosure.

Claims

1. An arylpiperidine derivative compound or a pharmaceutically acceptable salt thereof, as shown in Formula 1 below: [Chemical Formula 1] in, In the chemical formula 1, R 1 To R 3 They may be the same or different, and each is independently selected from hydrogen (H), nitro (NO2) and cyano (CN). R 4 For selection or either of them, E1 through E8 may be the same or different, and each is independently CH or N. R' is hydrogen (H) or a C1~C4 alkyl group, and n can be any number from 1 to 5.

2. The arylpiperidine derivative compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein, In the chemical formula 1, R 2 For hydrogen (H) or nitro (NO2), R 4 for , , and In any of the following, E1 is N, E3 to E5 and E8 are CH, R' is hydrogen (H) or methyl, and n is any one of 1 to 3.

3. The arylpiperidine derivative compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein, The arylpiperidine derivatives are selected from the group consisting of: N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-2-(1-(2-cyanophenyl)piperidin-4-yl)acetamide, 2-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-b]pyridazin-3-yl)acetamide, 2-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-a]pyridin-3-yl)acetamide, 2-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-a]pyridin-3-yl)acetamide, 2-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-a]pyridin-5-yl)acetyl Amines, N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(2-cyanophenyl)piperidin-4-yl)propionamide, N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-4-(1-(2-cyanophenyl)piperidin-4-yl)butyramide, 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-b]pyridazin-3-yl)propionamide, 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-a]pyridin-3-yl)propionamide, 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(imidazo[1,2-a]pyridin-3-yl)propionamide, 3-(1-(2-cyanophenyl) Piperidin-4-yl)-N-(imidazo[1,2-a]pyridin-5-yl)propionamide, 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(1-methyl-1H-indol-6-yl)propionamide, 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(1H-indol-4-yl)propionamide, 3-(1-(2-cyanophenyl)piperidin-4-yl)-N-(1H-indol-7-yl)propionamide, N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(2-nitrophenyl)piperidin-4-yl)propionamide, N-([ [1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(3-nitrophenyl)piperidin-4-yl)propamide, N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(4-nitrophenyl)piperidin-4-yl)propamide, N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(4-cyanophenyl)piperidin-4-yl)propamide, and N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3-(1-(2,4-dinitrophenyl)piperidin-4-yl)propamide.

4. The arylpiperidine derivative compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein, In the chemical formula 1, R 1 It is cyano (CN), R 2 and R 3 For hydrogen (H), R 4 for E1, E6 and E7 are N, E2 to E5 and E8 are CH, and n is 1 or 2.

5. An arylpiperidine derivative compound represented by the following chemical formula 2, or a pharmaceutically acceptable salt thereof: [Chemical Formula 2] in, In the chemical formula 2, R 1 To R 3 They may be the same or different, and each is independently selected from hydrogen (H) or cyano (CN). R 4 for or , E1 through E8 may be the same or different, and each is independently CH or N. L is NR' or NH2 + ,and R' is a C1~C4 alkyl or C1~C5 alkoxycarbonyl.

6. The arylpiperidine derivative compound of claim 5 or a pharmaceutically acceptable salt thereof, wherein, In the chemical formula 2, R 1 It is cyano (CN), R 2 and R 3 For hydrogen (H), R 4 for E1, E6 and E7 are N, E2 to E5 and E8 are CH, and R' is C1 to C2 alkyl or C3 to C4 alkoxycarbonyl.

7. The arylpiperidine derivative compound of claim 5 or a pharmaceutically acceptable salt thereof, wherein, The arylpiperidine derivatives are selected from the group consisting of: (2-([1,2,4]triazolo[4,3-a]pyridin-3-ylamino)-2-oxoethyl)(1-(2-cyanophenyl)piperidin-4-yl)carbamate tert-butyl ester, N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-2-((1-(2-cyanophenyl)piperidin-4-yl)amino)acetamide, and N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-2-((1-(2-cyanophenyl)piperidin-4-yl)(methyl)amino)acetamide.

8. The arylpiperidine derivative compound of claim 7 or a pharmaceutically acceptable salt thereof, wherein, The N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-2-((1-(2-cyanophenyl)piperidin-4-yl)amino)acetamide forms an ionic bond with any of the groups selected from the group consisting of: acetate (CH3COO) - ), trifluoroacetate (CF3COO) - ), chloride ions (Cl) - ), and methyl p-toluenesulfonate (CH3C6H4SO3) - ).

9. A pharmaceutical composition for treating or preventing end-anchor polymerase-related cancer, said pharmaceutical composition comprising an arylpiperidine derivative compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-8.

10. The pharmaceutical composition of claim 9, wherein, The cancers mentioned are selected from the group consisting of: colon cancer, lung cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, breast cancer, stomach cancer, liver cancer, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, breast cancer, brain cancer, laryngeal cancer, prostate cancer, bladder cancer, esophageal cancer, thyroid cancer, kidney cancer, leukemia, and rectal cancer.

11. A pharmaceutical composition for a combination therapy for treating end-anchor polymerase-related cancer, said pharmaceutical composition comprising an arylpiperidine derivative compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-8, and an anticancer agent.

12. The pharmaceutical composition of claim 11, wherein, The anticancer agents are selected from the group consisting of: 7-ethyl-10-hydroxycamptothecin, 5-fluorouracil, cisplatin, paclitaxel, doxorubicin, daunorubicin, vincristine, actinomycin D, teniposide, etoposide, cyclophosphamide, epirubicin, doxorubicin, doxorubicin, and mitomycin-C.

13. The pharmaceutical composition of claim 11, wherein, The cancers mentioned are selected from the group consisting of: colon cancer, lung cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, breast cancer, stomach cancer, liver cancer, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, breast cancer, brain cancer, laryngeal cancer, prostate cancer, bladder cancer, esophageal cancer, thyroid cancer, kidney cancer, leukemia, and rectal cancer.

14. A pharmaceutical composition for enhancing anticancer effects against end-anchor polymerase-related cancers, said pharmaceutical composition comprising an arylpiperidine derivative compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-8, and an anticancer agent.

15. The pharmaceutical composition of claim 14, wherein, The cancers mentioned are selected from the group consisting of: colon cancer, lung cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, breast cancer, stomach cancer, liver cancer, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, breast cancer, brain cancer, laryngeal cancer, prostate cancer, bladder cancer, esophageal cancer, thyroid cancer, kidney cancer, leukemia, and rectal cancer.

16. A health-functional food composition for improving or preventing end-anchor polymerase-related cancers, said health-functional food composition comprising an arylpiperidine derivative compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-8, and an anticancer agent.

17. The health functional food composition as described in claim 16, wherein, The cancers mentioned are selected from the group consisting of: colon cancer, lung cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, breast cancer, stomach cancer, liver cancer, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, breast cancer, brain cancer, laryngeal cancer, prostate cancer, bladder cancer, esophageal cancer, thyroid cancer, kidney cancer, leukemia, and rectal cancer.

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