Piperidine-2,6-dione derivative and use thereof

Abstract

The present disclosure relates to a novel piperidine-2,6-dione derivative and a use thereof and, more specifically, to a piperidine-2,6-dione derivative compound having a structure of a thalidomide analog. A compound of chemical formula 1 according to the present disclosure specifically binds with CRBN protein, and is involved in functions thereof. Therefore, the compound of the present disclosure can be favorably used in the prevention or treatment of leprosy, chronic graft versus host disease, an inflammatory disease, or cancer, which are caused by actions of CRBN protein.

Claims

1. A compound represented by the following Chemical formula 1: ##STR00053## or its pharmaceutically acceptable salt, in the Chemical formula 1, X is nitrogen (N) and R.sub.1 or R.sub.2 is each independently any one selected from the group consisting of hydrogen, hydroxy, halogen, cyano, amino, nitro, —NR.sub.5R.sub.6, —N(SO.sub.2)R.sub.7, —CONR.sub.5R.sub.6, —OR.sub.5, —SR.sub.5, —SO.sub.2R.sub.5, —SO.sub.2NR.sub.5R.sub.6, —CR.sub.5R.sub.6, —CR.sub.5NR.sub.6R.sub.7, —P(O)(OR.sub.5)R.sub.6, —P(O)R.sub.5R.sub.6, —OP(O)(OR.sub.5)R.sub.6, —OP(O)R.sub.5R.sub.6, —CF.sub.3, —NR.sub.5SO.sub.2NR.sub.5R.sub.6, —CONR.sub.5COR.sub.6, —NR.sub.5C(═N—CN)NR.sub.5R.sub.6, —C(═N—CN)NR.sub.5R.sub.6, —NR.sub.5C(═N—CN)R.sub.6, —NR.sub.5C(═C—NO.sub.2)NR.sub.5R.sub.6, —SO.sub.2NR.sub.5COR.sub.6, —CO.sub.2R.sub.5, —C(C═N—OR.sub.5)R.sub.6, —CR.sub.5═CR.sub.5R.sub.6, —CCR.sub.5, —S(C═O)(C═N—R.sub.5)R.sub.6, —SF.sub.5, —OCF.sub.3, —NHCOR.sub.5, C.sub.1-C.sub.10 linear or branched alkyl, C.sub.2-C.sub.10 linear or branched alkenyl, C.sub.2-C.sub.10 linear or branched alkynyl, C.sub.1-C.sub.10 linear or branched alkoxy, C.sub.3-C.sub.10 cycloalkyl, C.sub.3-C.sub.10 heterocycloalkyl comprising one or more heteroatoms selected from the group consisting of N, O and S, C.sub.3-C.sub.10 cycloalkenyl, C.sub.3-C.sub.10 heterocycloalkenyl comprising one or more heteroatoms selected from the group consisting of N, O and S, C.sub.6-C.sub.14 aryl, C.sub.6-C.sub.24 arylalkyl, and C.sub.6-C.sub.14 heteroaryl comprising one or more heteroatoms selected from the group consisting of N, O and S, and R.sub.3 is any one selected from the group consisting of hydrogen, deuterium and C.sub.1-C.sub.10 linear or branched alkyl, and R.sub.4 is any one selected from the group consisting of hydrogen, —(CH.sub.2).sub.nOCOR.sub.8 and C.sub.1-C.sub.10 linear or branched alkyl, and R.sub.5 to R.sub.7 are each independently any one selected from the group consisting of hydrogen, amino, C.sub.1-C.sub.10 linear or branched alkyl, C.sub.2-C.sub.10 linear or branched alkenyl, C.sub.2-C.sub.10 linear or branched alkynyl, C.sub.3-C.sub.10 cycloalkyl, C.sub.3-C.sub.10 heterocycloalkyl comprising one or more heteroatoms selected from the group consisting of N, O and S, C.sub.6-C.sub.14 aryl and C.sub.6-C.sub.14 heteroaryl comprising one or more heteroatoms selected from the group consisting of N, O and S, R.sub.8 is any one selected from the group consisting of hydrogen, unsubstituted or substituted amino, unsubstituted or substituted C.sub.1-C.sub.10 linear or branched alkyl, unsubstituted or substituted C.sub.2-C.sub.10 linear or branched alkenyl, unsubstituted or substituted C.sub.2-C.sub.10 linear or branched alkynyl, unsubstituted or substituted C.sub.3-C.sub.10 cycloalkyl, unsubstituted or substituted C.sub.3-C.sub.10 heterocycloalkyl comprising one or more heteroatoms selected from the group consisting of N, O and S, unsubstituted or substituted C.sub.6-C.sub.14 aryl or unsubstituted or substituted C.sub.6-C.sub.14 heteroaryl comprising one or more heteroatoms selected from the group consisting of N, O and S, wherein said substituted amino, substituted C.sub.1-C.sub.10 alkyl, substituted C.sub.2-C.sub.10 alkenyl, substituted C.sub.2-C.sub.10 alkynyl, substituted C.sub.3-C.sub.10 cycloalkyl, substituted C.sub.3-C.sub.10 heterocycloalkyl, substituted C.sub.6-C.sub.14 aryl, and substituted C.sub.6-C.sub.14 heteroaryl wherein the substituted R.sub.8 are substituted with one or more substituents selected from the group consisting of halogen atom, nitro, hydroxy, cyano, amino, thiol, carboxyl, amide, nitrile, sulfide, disulfide, sulphenyl, formyl, formyloxy, formylamino, C.sub.6-C.sub.14 aryl, and tert-butyloxycarbonyl (BOC), and n is an integer of 1 to 5.

2. The compound or its pharmaceutically acceptable salt according to claim 1, wherein in the Chemical formula 1, R.sub.1 or R.sub.2 is each independently any one selected from the group consisting of hydrogen, hydroxy, halogen, cyano, amino, nitro, —CH.sub.3, —OCF.sub.3, —OCH.sub.3, —NHCOCH.sub.3, —COOH, —CONH.sub.2, —CONHCH.sub.3 and —NHCOC.sub.6H.sub.5, and R.sub.4 is any one selected from the group consisting of ##STR00054##

3. The compound or its pharmaceutically acceptable salt according to claim 1, wherein the compound of Chemical formula 1 is selected from the group consisting of 3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, 3-(5-fluoro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, 3-(5-nitro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, 3-(5-chloro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, 3-(5-methyl-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, 3-(6-chloro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, 3-(6-bromo-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, 3-(6-iodo-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, 3-(8-bromo-6-methyl-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, 3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-6-carbonitrile, 3-(5-iodo-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, 3-(4-oxo-6-(trifluoromethoxy)benzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, 3-(6-methoxy-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, N-(3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-5-yl)acetamide, 3-(8-nitro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, 3-(8-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, N-(3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-6-yl)acetamide, 3-(4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, 3-(7-nitro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, 3-(6-nitro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, 3-(7-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, 3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-5-carboxylic acid, 3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)-3-methylpiperidine-2,6-dione, 3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-5-carboxamide, 3-(2,6-dioxopiperidin-3-yl)-N-methyl-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-5-carboxamide, N-(3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-5-yl)benzamide, 3-(5-hydroxy-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione, (3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)-2,6-dioxopiperidin-1-yl)methyl pivalate, (3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)-2,6-dioxopiperidin-1-yl)methyl benzoate, (3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)-2,6-dioxopiperidin-1-yl)methyl piperidine-4-carboxylate hydrochloride, (3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)-2,6-dioxopiperidin-1-yl)methyl D-prolinate hydrochloride, (3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)-2,6-dioxopiperidin-1-yl)methyl L-isoleucinate hydrochloride, (3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)-2,6-dioxopiperidin-1-yl)methyl butyrate, 2-((3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)-2,6-dioxopiperidin-1-yl)methyl) 1-(tert-butyl) (2R)-pyrrolidine-1,2-dicarboxylate and 3-(6-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione.

4. A method comprising (a) reacting the compound represented by the following Chemical formula 2 with the compound represented by the following Chemical formula 3 to prepare a compound represented by the following Chemical formula 4; and (b) reacting the compound represented by the following Chemical formula 4 with NaNO.sub.2 to prepare the compound represented by the following Chemical formula 5, ##STR00055## in the Chemical formula 2, R.sub.1 or R.sub.2 are each independently any one selected from the group consisting of hydrogen, hydroxy, halogen, cyano, amino, nitro, —CH.sub.3, —OCF.sub.3, —OCH.sub.3, —NHCOCH.sub.3, —COOH, —CONH.sub.2, —CONHCH.sub.3 and —NHCOC.sub.6H.sub.5.

5. A method for treating leprosy, chronic graft versus host disease, or cancer, comprising: administering to a subject in need thereof a therapeutically effective amount of the compound or its pharmaceutically acceptable salt of claim 1, wherein the cancer is selected from the group consisting of breast cancer, colorectal cancer, lung cancer, small cell lung cancer, gastric cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, anal region cancer, colon cancer, breast cancer, fallopian tube carcinoma, endometrial cancer, cervical cancer, vaginal cancer, vulva cancer, Hodgkin disease, esophageal cancer, small intestine cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, CNS tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma and pituitary adenoma.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is the measurement of the degradation activity against Aiolos when the compound of Chemical formula 7 of the present disclosure is treated for 6 hours.

(2) FIG. 2 is the measurement of the degradation activity against GSPT1 and Aiolos when the compound of Chemical formula 37 or 38 of the present disclosure is treated for 6 hours or 24 hours.

DETAILED DESCRIPTION

(3) Hereinafter, the present disclosure will be described in detail.

(4) However, the following examples are intended to exemplify the present disclosure only, but the content of the present disclosure is not limited by the following examples.

Example 1

Synthesis of 3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 7)

(5) ##STR00017##

[1-1] Synthesis of 2-amino-N-(2,6-dioxopiperidin-3-yl)-6-fluorobenzamide (Compound <1-2>)

(6) 2-amino-6-fluorobenzoic acid (394 mg, 2.54 mmol) was dissolved in DMF (12 ml) and EDCl-HCl (540 mg, 2.8 mmol) and HOBt (429 mg, 2.8 mmol) were added. After stirring at a room temperature for 30 minutes, amine (361 mg, 2.8 mmol) was added, and DIPEA (1.4 ml, 8.12 mmol) was added, and it was stirred at a room temperature for 16 hours. The reaction mixture was diluted with water and then was extracted with ethyl acetate. The organic layer was dried on Na.sub.2SO.sub.4, and it was purified by column chromatography, to obtain pure 2-amino-N-(2,6-dioxopiperidin-3-yl)-6-fluorobenzamide compound 406 mg (60%) as a white solid.

(7) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 10.90 (s, 1H), 8.53 (d, J=6.0 Hz, 1H), 7.09 (dd, J=14.9, 8.1 Hz, 1H), 6.52 (d, J=8.2 Hz, 1H), 6.42-6.26 (m, 1H), 6.00 (s, 2H), 4.84-4.61 (m, 1H), 2.97-2.70 (m, 1H), 2.56-2.54 (m, 1H), 2.24-1.94 (m, 2H);

(8) MS found (M+H).sup.+ (m/z), 266.09; calcd for C.sub.12H.sub.12FN.sub.3O.sub.3 m/z, 266.10.

[1-2] Synthesis of 3-(5-fluoro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 8)

(9) Sodium nitride (179 mg, 2.6 mmol) was added to 2-amino-N-(2,6-dioxopiperidin-3-yl)-6-fluorobenzamide (406 mg, 1.5 mmol) dissolved in acetic acid (12 ml). After stirring at a room temperature for 1.5 hours, the mixture was diluted with water, and the precipitated white products were collected and were washed with water and were dried to obtain 3-(5-fluoro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione 280 mg.

(10) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.21 (s, 1H), 8.26-8.02 (m, 2H), 8.00-7.67 (m, 1H), 6.06-5.81 (m, 1H), 3.08-2.53 (m, 3H), 2.36-2.17 (m, 1H);

(11) MS found (M+H).sup.+ (m/z), 277.09; calcd for C.sub.12H.sub.9FN.sub.4O.sub.3 m/z, 277.10.

[1-3] Synthesis of 3-(5-((4-methoxybenzyl)amino)-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Compound <1-3>)

(12) To the solution of 3-(5-fluoro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (50 mg, 0.18 mmol) dissolved in DMF (1 ml), 4-methoxybenzyl amine (47 μl, 0.36 mmol) and DIPEA (62 μl, 0.36 mmol) were added at a room temperature, and it was stirred at 90° C. overnight. The reaction mixture was diluted with water, and then it was extracted with ethyl acetate. The organic layer was dried on anhydrous Na.sub.2SO.sub.4, and it was purified by column chromatography to obtain 3-(5-((4-methoxybenzyl)amino)-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (12 mg, 17%) as a white solid.

(13) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.55 (s, 1H), 8.25 (s, 1H), 7.64 (t, J=8.1 Hz, 1H), 7.29-7.27 (m, 2H), 6.88 (d, J=8.4 Hz, 2H), 6.76 (d, J=8.4 Hz, 1H), 5.76-5.61 (m, 1H), 4.39 (d, J=5.3 Hz, 2H), 3.80 (s, 3H), 3.00-2.71 (m, 3H), 2.49-2.28 (m, 1H);

(14) MS found (M+H).sup.+ (m/z), 394.9; calcd for C.sub.20H.sub.19N.sub.5O.sub.4 m/z, 394.10.

[1-4] Synthesis of 3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione

(15) To 3-(5-((4-methoxybenzyl)amino)-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione solution (10 mg, 0.025 mmol) dissolved in toluene (1 ml), DDQ (6.9 mg, 0.03 mmol) was added at a room temperature, and it was stirred overnight. The reaction mixture was diluted with ethyl acetate and was washed with NaHCO.sub.3 (aq). The organic layer was dried on anhydrous MgSO.sub.4, and it was purified by column chromatography to obtain a pure compound, 3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione 1.6 mg (23%) as a white solid.

(16) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.00 (s, 1H), 7.72-7.57 (m, 1H), 7.36 (d, J=7.8 Hz, 2H), 7.01-6.75 (m, 2H), 6.13 (s, 1H), 5.83-5.61 (m, 1H), 3.10-2.67 (m, 3H), 2.46-2.34 (m, 1H);

(17) MS found (M+H).sup.+ (m/z), 274.9; calcd for C.sub.12H.sub.11N.sub.5O.sub.3 m/z, 274.10.

Example 2

Synthesis of 3-(5-nitro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 9)

(18) ##STR00018##

[2-1] Synthesis of 2-amino-N-(2,6-dioxopiperidin-3-yl)-6-nitrobenzamide (Compound <2-2>)

(19) To 2-amino-6-nitrobenzoic acid (7 g, 38 mmol) dissolved in DMF (90 ml), EDCl-HCl (8 g, 42 mmol) and HOBt (6.5 g, 42 mmol) were added. After stirring at a room temperature for 30 minutes, 3-aminopiperidine-2,6-dione hydrochloride (25 g, 152 mmol) and DIPEA (21 ml, 121.6 mmol) were added and it was stirred at a room temperature for 16 hours. The reaction mixture was diluted with water and it was extracted with ethyl acetate. The organic layer was dried on anhydrous Na.sub.2SO.sub.4, and then was concentrated under the reduced pressure to obtain 2-amino-N-(2,6-dioxopiperidin-3-yl)-6-nitrobenzamide as a yellow solid (15 g), and it was used in the following step without purification.

(20) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.01 (s, 1H), 9.03 (d, J=8.3 Hz, 1H), 7.38-7.14 (m, 2H), 7.06 (d, J=7.2 Hz, 1H), 6.02 (s, 2H), 4.80-4.65 (m, 1H), 2.89-2.58 (m, 2H), 2.33-1.87 (m, 2H).

[2-2] Synthesis of 3-(5-nitro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione

(21) Sodium nitride (2.7 g, 40 mmol) was added to the solution of 2-amino-N-(2,6-dioxopiperidin-3-yl)-6-nitrobenzamide dissolved in acetic acid. After stirring at a room temperature for 1.5 hours, the mixture was diluted with water, and the precipitated white products were collected, and were washed using water, and were dried to obtain 3-(5-nitro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione 7 g (60%, step 2 yield).

(22) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.26 (s, 1H), 8.52 (dd, J=7.6, 1.5 Hz, 1H), 8.45-8.20 (m, 2H), 6.00 (dd, J=12.4, 5.3 Hz, 1H), 3.04-2.87 (m, 1H), 2.78-2.58 (m, 2H), 2.44-2.17 (m, 1H).

[2-3] Synthesis of 3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione

(23) To the solution of 3-(8-nitro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (1 g, 3.2 mmol) dissolved in AcOH/THF (25 ml/25 ml), iron (Fe, 1 g) was added and it was stirred at a room temperature overnight. The reaction mixture was filtered and extracted with ethyl acetate. The organic layer was dried on Na.sub.2SO.sub.4, and it was purified by column chromatography to obtain 0.5 g (57%) of pure 3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione as a yellow-green solid.

(24) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.00 (s, 1H), 7.72-7.57 (m, 1H), 7.36 (d, J=7.8 Hz, 2H), 7.01-6.75 (m, 2H), 6.13 (s, 1H), 5.83-5.61 (m, 1H), 3.10-2.67 (m, 3H), 2.46-2.34 (m, 1H);

(25) MS found (M+H).sup.+ (m/z), 274.9; calcd for C.sub.12H.sub.11N.sub.5O.sub.3 m/z, 274.10.

Example 3

Synthesis of 3-(5-chloro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 10)

(26) ##STR00019##

[3-1] Synthesis of 5-chloro-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (Compound <3-2>)

(27) To the solution of 2-amino-6-chlorobenzoic acid (1 g, 5.8 mmol) dissolved in 1,4-dioxane (10 ml), triphosgene (570 mg, 1.9 mmol) was added, and the solution was refluxed for 2 hours. Then, the reactant was cooled on ice. The solid was washed with hexane and was under vacuum drying to obtain a 1.1 g (99%) of pure compound as a brown solid.

(28) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ .sup.1H NMR (300 MHz, Methanol-d.sub.4) δ 7.63 (t, J=8.1 Hz, 1H), 7.40-7.24 (m, 1H), 7.10 (d, J=8.2 Hz, 1H);

(29) MS found (M+H).sup.+ (m/z), 198.09; calcd for C.sub.8H.sub.4ClNO.sub.3 m/z, 198.10.

[3-2] Synthesis of 2-amino-6-chloro-N-(2,6-dioxopiperidin-3-yl)benzamide (Compound <3-3>)

(30) To the solution of 5-chloro-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (200 mg, 1.01 mmol) dissolved in DMF (4 ml), 3-aminopiperidine-2,6-dione (194 mg, 1.5 mmol) and DMAP (12 mg, 0.10 mmol) were added, and the temperature was elevated from 60° C. overnight. The reaction mixture was extracted with ethyl acetate and the organic layer was dried on Na.sub.2SO.sub.4. The crude compound was obtained as a blue solid (75 mg), and this was used in the next step without further purification.

(31) MS found (M+H).sup.+ (m/z), 282.09; calcd for C.sub.12H.sub.9ClN.sub.4O.sub.3 m/z, 282.10.

[3-3] Synthesis of 3-(5-chloro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione

(32) To the solution of 2-amino-6-chloro-N-(2,6-dioxopiperidin-3-yl)benzamide (75 mg, 0.26 mmol) dissolved in acetic acid (2 ml), NaNO.sub.2 (31 mg, 0.45 mmol) was added, and it was stirred at a room temperature for 1 hour. The reaction mixture was diluted with water and the precipitated product was collected to obtain a pure compound of 18 mg (24%) as a white solid.

(33) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.22 (s, 1H), 8.21 (d, J=7.9 Hz, 1H), 8.12-8.01 (m, 2H), 6.02-5.88 (m, 1H), 3.03-2.88 (m, 1H), 2.84-2.60 (m, 2H), 2.42-2.14 (m, 1H);

(34) MS found (M+H).sup.+ (m/z), 293.09; calcd for C.sub.12H.sub.9ClN.sub.4O.sub.3 m/z, 293.10.

Example 4

Synthesis of 3-(5-methyl-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 11)

(35) ##STR00020##

[4-1] Synthesis of 5-methyl-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (Compound <4-2>)

(36) To the solution of 2-amino-6-methylbenzoic acid (876 mg, 5.8 mmol) dissolved in 1,4-dioxane (10 ml), triphosgene (570 mg, 1.9 mmol) was added, and the solution was refluxed for 2 hours. Then, the solution was cooled on ice. The solid was washed with hexane and was under vacuum drying to obtain a pure compound of 880 mg (85%) as a brown solid.

(37) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 9.02 (brs, 1H), 7.56 (t, J=7.8 Hz, 1H), 7.12 (d, J=7.7 Hz, 1H), 6.94 (d, J=8.1 Hz, 1H), 2.77 (s, 3H).

[4-2] Synthesis of 2-amino-N-(2,6-dioxopiperidin-3-yl)-6-methylbenzamide (Compound <4-3>)

(38) To the solution of 5-methyl-2H-benzo[d][1,3]oxazine-2,4[1h]-dione (178 mg, 1.01 mmol) dissolved in DMF (4 ml), 3-aminopiperidine-2,6-dione (194 mg, 1.5 mmol) and DMAP (12 mg, 0.104 mmol) were added, and the mixture was heated at 60° C. overnight. The reaction mixture was extracted with ethyl acetate, and the organic layer was dried on Na.sub.2SO.sub.4. The crude compound was obtained as a blue solid (50 mg), and this was used in the next step without further purification.

[4-3] Synthesis of 3-(5-methyl-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione

(39) To the solution of 2-amino-N-(2,6-dioxopiperidin-3-yl)-6-methylbenzamide (50 mg, 0.19 mmol) dissolved in acetic acid (2 ml), NaNO.sub.2 (22 mg, 0.32 mmol) was added, and it was stirred at a room temperature for 1 hour. The reaction mixture was diluted with water, and the precipitated product was collected, and a pure compound of 16 mg (31%) was obtained as a brown solid.

(40) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.19 (s, 1H), 8.07 (d, J=7.8 Hz, 1H), 7.99 (t, J=7.7 Hz, 1H), 7.75 (d, J=7.1 Hz, 1H), 5.99-5.87 (m, 1H), 3.03-2.85 (m, 1H), 2.83 (s, 3H), 2.76-2.56 (m, 2H), 2.35-2.19 (m, 1H).

Example 5

Synthesis of 3-(6-chloro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 12)

(41) ##STR00021##

[5-1] Synthesis of 6-chloro-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (Compound <5-2>)

(42) To the solution of 2-amino-5-chlorobenzoic acid (500 mg, 2.0 mmol) dissolved in 1,4-dioxane (5 ml), triphosgene (285 mg, 0.96 mmol) was added and the solution was refluxed for 2 hours. The reaction mixture was cooled in an ice container. The solid was washed and was under vacuum drying to obtain a pure compound of 400 mg (70%) as a brown solid.

(43) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.88 (s, 1H), 7.89 (d, J=2.4 Hz, 1H), 7.80 (dd, J=8.7, 2.5 Hz, 1H), 7.19 (d, J=8.7 Hz, 1H).

[5-2] Synthesis of 2-amino-5-chloro-N-(2,6-dioxopiperidin-3-yl)benzamide (Compound <5-3>)

(44) To the solution of 6-chloro-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (200 mg, 1.01 mmol) dissolved in DMF (3 ml), 3-aminopiperidine-2,6-dione (129 mg, 1.01 mmol) and DMAP (12.3 mg, 0.101 mmol) were added, and the reaction mixture solution was heated at 60° C. overnight. The reaction mixture was extracted with ethyl acetate, and the organic layer was dried on Na.sub.2SO.sub.4. The crude compound was obtained as a blue solid (70 mg), and this was used in the next step without further purification.

[5-3] Synthesis of 3-(6-chloro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione

(45) To the solution of 2-amino-N-(2,6-dioxopiperidin-3-yl)-6-methylbenzamide (50 mg, 0.19 mmol) dissolved in acetic acid (3 ml), NaNO.sub.2 (22 mg, 0.32 mmol) was added and it was stirred at a room temperature for 1 hour. The reaction mixture was diluted with water, and the precipitated product was collected to obtain a pure compound of 28 mg (39%) as a brown solid.

(46) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.21 (br s, 1H), 8.38-8.25 (m, 2H), 8.24-8.12 (m, 1H), 6.00 (dd, J=12.2, 5.4 Hz, 1H), 3.05-2.91 (m, 1H), 2.83-2.58 (m, 2H), 2.39-2.15 (m, 2H).

Example 6

Synthesis of 3-(6-bromo-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 13)

(47) ##STR00022##

[6-1] Synthesis of 6-bromo-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (Compound <6-2>)

(48) To the solution of 2-amino-5-bromobenzoic acid (626 mg, 2.9 mmol) dissolved in 1,4-dioxane (5 ml), triphosgene (285 mg, 0.96 mmol) was added, and the solution was refluxed for 2 hours. Then, the reaction mixture was cooled on ice. The solid was washed with hexane and was under vacuum drying to obtain a pure compound of brown solid of 630 mg (90%).

(49) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.87 (s, 1H), 8.00 (d, J=2.3 Hz, 1H), 7.91 (dd, J=8.7, 2.3 Hz, 1H), 7.11 (d, J=8.7 Hz, 1H).

[6-2] Synthesis of 2-amino-5-bromo-N-(2,6-dioxopiperidin-3-yl)benzamide (Compound <6-3>)

(50) To the solution of 6-bromo-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (244 mg, 1.01 mmol) dissolved in DMF (3 ml), 3-aminopiperidine-2,6-dione (129 mg, 1.01 mmol) and DMAP (12.3 mg, 0.101 mmol) were added, and the mixture was heated at 60° C. overnight. The reaction mixture was extracted with ethyl acetate, and the organic layer was dried on Na.sub.2SO.sub.4. The crude compound was obtained as a blue solid (80 mg), and this was used in the next step without further purification.

[6-3] Synthesis of 3-(6-bromo-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione

(51) To the solution of 2-amino-5-bromo-N-(2,6-dioxopiperidin-3-yl)benzamide (80 mg, 0.25 mmol) dissolved in acetic acid (3 ml), NaNO.sub.2 (28 mg, 0.41 mmol) was added, and it was stirred at a room temperature for 1 hour. The reaction mixture was diluted with water, and the precipitated product was collected to obtain a pure compound of 42 mg (51%) as an ivory solid.

(52) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.21 (br s, 1H), 8.40 (d, J=2.0 Hz, 1H), 8.32 (dd, J=8.6, 2.2 Hz, 1H), 8.21 (d, J=8.6 Hz, 1H), 6.07-5.99 (m, 1H), 3.00-2.85 (m, 1H), 2.78-2.55 (m, 2H), 2.34-2.16 (m, 1H).

Example 7

Synthesis of 3-(6-iodo-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 14)

(53) ##STR00023##

[7-1] Synthesis of 6-iodo-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (Compound <7-2>)

(54) To the solution of 2-amino-5-iodobenzoic acid (762 mg, 2.9 mmol) dissolved in 1,4-dioxane (5 ml), triphosgene (285 mg, 0.96 mmol) was added, and the solution was refluxed for 2 hours. Then, the reaction mixture was cooled on ice. The solid was washed with hexane and was under vacuum drying to obtain a pure compound of brown solid of 680 mg (81%).

(55) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.83 (s, 1H), 8.14 (d, J=2.0 Hz, 1H), 8.03 (dd, J=8.5, 2.0 Hz, 1H), 6.97 (d, J=8.6 Hz, 1H).

[7-2] Synthesis of 2-amino-N-(2,6-dioxopiperidin-3-yl)-5-iodobenzamide (Compound <7-3>)

(56) To the solution of 6-iodo-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (291 mg, 1.01 mmol) dissolved in DMF (3 ml), 3-aminopiperidine-2,6-dione (129 mg, 1.01 mmol) and DMAP (12.3 mg, 0.101 mmol) were added, and the mixture was heated at 60° C. overnight. The reaction mixture was extracted with ethyl acetate, and the organic layer was dried on Na.sub.2SO.sub.4. The crude compound was obtained as a blue solid (80 mg), and this was used in the next step without further purification.

[7-3] Synthesis of 3-(6-iodo-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione

(57) To the solution of 2-amino-N-(2,6-dioxopiperidin-3-yl)-5-iodobenzamide (130 mg, 0.35 mmol) dissolved in acetic acid (5 ml), NaNO.sub.2 (40 mg, 0.59 mmol) was added, and it was stirred at a room temperature for 1 hour. The reaction mixture was diluted with water, and the precipitated product was collected to obtain a pure compound of 20 mg (15%) as an ivory solid.

(58) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.20 (br s, 1H), 8.58 (s, 1H), 8.47 (d, J=8.3 Hz, 1H), 8.02 (d, J=8.6 Hz, 1H), 6.14-5.84 (m, 1H), 3.02-2.83 (m, 1H), 2.85-2.63 (m, 2H), 2.40-2.01 (m, 1H).

Example 8

Synthesis of 3-(8-bromo-6-methyl-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 15)

(59) ##STR00024##

[8-1] Synthesis of 8-bromo-6-methyl-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (Compound <8-2>)

(60) To the solution of 2-amino-3-bromo-5-methylbenzoic acid (667 mg, 2.9 mmol) dissolved in 1,4-dioxane (5 ml), triphosgene (285 mg, 0.96 mmol) was added, and the solution was refluxed for 2 hours. Then, the reaction mixture was cooled on ice. The solid was washed with hexane and was under vacuum drying to obtain a pure compound of brown solid of 720 mg (97%).

(61) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.01 (s, 1H), 7.89 (d, J=1.4 Hz, 1H), 7.77 (s, 1H), 3.57 (s, 3H).

[8-2] Synthesis of 2-amino-3-bromo-N-(2,6-dioxopiperidin-3-yl)-5-methylbenzamide (Compound <8-3>)

(62) To the solution of 8-bromo-6-methyl-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (258 mg, 1.01 mmol) dissolved in DMF (3 ml), 3-aminopiperidine-2,6-dione (129 mg, 1.01 mmol) and DMAP (12.3 mg, 0.101 mmol) were added, and the mixture was heated at 60° C. overnight. The reaction mixture was extracted with ethyl acetate, and the organic layer was dried on Na.sub.2SO.sub.4. The crude compound was obtained as a blue solid (170 mg), and this was used in the next step without further purification.

[8-3] Synthesis of 3-(8-bromo-6-methyl-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione

(63) To the solution of 2-amino-3-bromo-N-(2,6-dioxopiperidin-3-yl)-5-methylbenzamide (170 mg, 0.5 mmol) dissolved in acetic acid (6 ml), NaNO.sub.2 (59 mg, 0.85 mmol) was added, and it was stirred at a room temperature for 1 hour. The reaction mixture was diluted with water, and the precipitated was collected to obtain a pure compound of 120 mg (69%) as an ivory solid.

(64) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.21 (br s, 1H), 8.31 (d, J=1.3 Hz, 1H), 8.13-8.03 (m, 1H), 6.00 (dd, J=12.1, 5.3 Hz, 1H), 2.99-2.91 (m, 1H), 2.78-2.59 (m, 1H), 2.55 (s, 2H), 2.40-2.21 (m, 1H).

Example 9

Synthesis of 3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-6-carbonitrile (Chemical Formula 16)

(65) ##STR00025##

(66) 3-(6-Iodo-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (10 mg, 0.038 mmol, synthesized in Example 7) dissolved in propionitrile (0.04 ml) was added, and it was stirred using a microwave reactor for 1 hour. The mixture was filtered with DCM, and was concentrated, and was purified by column chromatography to obtain a pure compound of 4.1 mg (40%) as a pink solid.

(67) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.71 (s, 1H), 8.35 (d, J=8.4 Hz, 1H), 8.22 (dd, J=8.4, 1.5 Hz, 1H), 8.12 (s, 1H), 5.88 (dd, J=12.2, 5.3 Hz, 1H), 3.15-2.79 (m, 3H), 2.60-2.39 (m, 1H).

Example 10

Synthesis of 3-(5-iodo-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 17)

(68) ##STR00026##

(69) To the solution of 3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (10 mg, 0.036 mmol, synthesized in Example 1) dissolved in concentrated hydrochloric acid of 0.12 ml, NaNO.sub.2 (5 mg, 0.073 mmol) comprised in 0.02 ml water was added at 0° C. under the presence of argon gas. After the addition, the solution was stirred for 30 minutes and KI (potassium iodide, 12 mg, 0.073 mmol) dissolved in 10 ml water was added at 0° C. under the presence of argon gas. Then, it was stirred overnight. The solution was diluted with 100 ml AcOEt, and was diluted again with 100 ml water. The aqueous layer was separated and was extracted with EtOAc. The organic layer was combined and it was washed with salt water, and it was dried on Na.sub.2SO.sub.4, and was concentrated under vacuum. The residues were purified by chromatography on gel to obtain 3-(5-iodo-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione.

(70) .sup.1H NMR (300 MHz, MeOD) δ 8.50 (d, J=7.7 Hz, 1H), 8.20 (d, J=8.2 Hz, 1H), 7.69 (t, J=7.5 Hz, 1H), 6.04-5.82 (m, 1H), 3.04-2.75 (m, 3H), 2.49-2.21 (m, 1H).

Example 11

Synthesis of 3-(4-oxo-6-(trifluoromethoxy)benzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 18)

(71) ##STR00027##

[11-1] Synthesis of 6-(trifluoromethoxy)-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (Compound <11-2>)

(72) To the solution of 2-amino-5-(trifluoromethoxy)benzoic acid (0.2 g, 0.904 mmol) dissolved in 1,4-dioxane (5 ml), triphosgene (0.09 g, 0.3 mmol) was added, and the solution was refluxed for 2 hours. The reaction mixture was cooled in an ice container. The solid was washed with hexane and was under vacuum drying to obtain a pure compound of 0.2 g (90%) as an ivory solid.

(73) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.92 (s, 1H), 7.82-7.76 (m, 2H), 7.25 (d, J=8.8 Hz, 1H).

[11-2] Synthesis of 2-amino-N-(2,6-dioxopiperidin-3-yl)-5-(trifluoromethoxy)benzamide (Compound <11-3>)

(74) To 6-(trifluoromethoxy)-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (0.2 g, 0.81 mmol) dissolved in DMF (2 ml), 3-aminopiperidine-2,6-dione (0.156 g, 1.22 mmol) and DMAP (0.015 g, 0.122 mmol) were added and the mixture was heated at 60° C. overnight. The reaction mixture was extracted with ethyl acetate, and the organic layer was dried on Na.sub.2SO.sub.4. The crude compound was collected as a blue solid (0.2 g), and this was used in the next step without further purification.

[11-3] Synthesis of 3-(4-oxo-6-(trifluoromethoxy)benzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione

(75) To the solution of 2-amino-N-(2,6-dioxopiperidin-3-yl)-5-(trifluoromethoxy)benzamide (0.1 g, 0.32 mmol) dissolved in acetic acid (2 ml), NaNO.sub.2 (37 mg, 0.54 mmol) was added, and it was stirred at a room temperature for 1 hour. The reaction mixture was diluted with water, and the precipitated product was collected to obtain a pure compound of 76 mg (70%) as an ivory solid.

(76) .sup.1H NMR (500 MHz, CD.sub.3OD) δ 8.36 (d, J=8.9 Hz, 1H), 8.15 (s, 1H), 7.99 (d, J=8.8 Hz, 1H), 6.01-5.97 (m, 1H), 2.89-2.83 (m, 3H), 2.41-2.37 (m, 1H);

(77) MS found (M+H).sup.+ (m/z), 342.8; calcd for C.sub.13H.sub.9F.sub.3N.sub.4O.sub.4 m/z, 342.06.

Example 12

Synthesis of 3-(6-methoxy-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 19)

(78) ##STR00028##

[12-1] Synthesis of 6-methoxy-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (Compound <12-2>)

(79) To the solution of 2-amino-5-methoxybenzoic acid (0.5 g, 3 mmol) dissolved in 1,4-dioxane (5 ml), triphosgene (0.3 g, 1 mmol) was added, and the solution was refluxed for 2 hours. The reaction mixture was cooled in an ice container. The solid was washed with hexane and was under vacuum drying to obtain a pure compound of 0.53 g (92%) as a brown solid.

(80) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.61 (s, 1H), 7.38 (dd, J=8.8, 2.9 Hz, 1H), 7.34 (d, J=2.8 Hz, 1H), 7.11 (d, J=8.8 Hz, 1H), 3.80 (s, 3H).

[12-2] Synthesis of 2-amino-N-(2,6-dioxopiperidin-3-yl)-5-methoxybenzamide (Compound <12-3>)

(81) To 6-methoxy-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (0.2 g, 1.04 mmol) dissolved in DMF (2 ml), 3-aminopiperidine-2,6-dione (0.2 g, 1.56 mmol) and DMAP (0.013 g, 0.104 mmol) were added, and the mixture was heated at 60° C. overnight. The reaction mixture was extracted with ethyl acetate, and the organic layer was dried on Na.sub.2SO.sub.4. The crude compound was obtained as a blue solid (0.2 g) and this was used in the next step without further purification.

[12-3] Synthesis of 3-(6-methoxy-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione

(82) To the solution of 2-amino-N-(2,6-dioxopiperidin-3-yl)-5-methoxybenzamide (0.13 g, 0.5 mmol) dissolved in acetic acid (2 ml), NaNO.sub.2 (60 mg, 0.85 mmol) was added, and it was stirred at a room temperature for 1 hour. The reaction mixture was diluted with water and the precipitated product was collected to obtain a pure compound of 42 mg (30%) as a pink solid.

(83) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.20 (s, 1H), 8.20 (d, J=8.9 Hz, 1H), 7.69 (dd, J=8.9, 2.9 Hz, 1H), 7.61 (d, J=2.8 Hz, 1H), 6.00-5.94 (m, 1H), 3.04-2.91 (m, 1H), 2.77-2.63 (m, 2H), 2.31-2.23 (m, 1H);

(84) MS found (M+H).sup.+ (m/z), 288.9; calcd for C.sub.13H.sub.12N.sub.4O.sub.4 m/z, 288.09.

Example 13

Synthesis of N-(3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-5-yl)acetamide (Chemical Formula 20)

(85) ##STR00029##

(86) To the solution of 3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (22 mg, 0.081 mmol, synthesized in Example 1) dissolved in DCM (5 ml), trimethylamine (0.013 ml, 0.09 mmol) and acetyl chloride (0.006 ml, 0.081 mmol) were added at 0° C., and the mixture was stirred at a room temperature overnight. The reaction mixture was extracted with DCM. The organic layer was dried on Na.sub.2SO.sub.4, and was purified by column chromatography to obtain a pure compound of 0.012 g (47%) as a white solid.

(87) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 11.46 (s, 1H), 9.06 (d, J=8.5 Hz, 1H), 8.06 (s, 1H), 7.96 (t, J=8.2 Hz, 1H), 7.86 (d, J=8.1 Hz, 1H), 5.76-5.71 (m, 1H), 3.03-2.84 (m, 1H), 2.44-2.37 (m, 1H), 2.28 (s, 3H);

(88) MS found (M+H).sup.+ (m/z), 315.9; calcd for C.sub.14H.sub.13N.sub.5O.sub.4 m/z, 315.10.

Example 14

Synthesis of 3-(8-nitro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 21) and 3-(8-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 22)

(89) ##STR00030##

[14-1] Synthesis of 8-nitro-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (Compound <14-2>)

(90) To the solution of 2-amino-3-nitrobenzoic acid (1.0 g, 5.5 mmol) dissolved in 1,4-dioxane (10 ml), triphosgene (0.54 g, 1.82 mmol) was added and the solution was refluxed overnight. The reaction mixture was cooled in an ice container. The solid was washed with hexane and was under vacuum drying to obtain a pure compound of 0.945 g (83%) as a yellow solid.

(91) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.13 (s, 1H), 8.49 (d, J=8.2 Hz, 1H), 8.34 (d, J=6.6 Hz, 1H), 7.43 (t, J=8.0 Hz, 1H).

[14-2] Synthesis of 2-amino-N-(2,6-dioxopiperidin-3-yl)-3-nitrobenzamide (Compound <14-3>)

(92) To 8-nitro-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (0.216 g, 1.04 mmol) dissolved in DMF (2 ml), 3-aminopiperidine-2,6-dione (0.2 g, 1.56 mmol) and DMAP (0.013 g, 0.104 mmol) were added, and the mixture was heated at 90° C. overnight. The reaction mixture was extracted with ethyl acetate, and the organic layer was dried on Na.sub.2SO.sub.4. The crude compound was obtained as a yellow solid (0.2 g), and this was used in the next step without further purification.

[14-3] Synthesis of 3-(8-nitro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione

(93) To the solution of 2-amino-N-(2,6-dioxopiperidin-3-yl)-3-nitrobenzamide (0.128 g, 0.46 mmol) dissolved in acetic acid (2 ml), NaNO.sub.2 (54 mg, 0.78 mmol) was added, and it was stirred at a room temperature for 1 hour. The reaction mixture was diluted with water, and the precipitated product was collected to obtain a pure compound of 55 mg (39%) as a yellow solid.

(94) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.26 (s, 1H), 8.64 (d, J=7.8 Hz, 1H), 8.53 (d, J=8.2 Hz, 1H), 8.14 (t, J=8.0 Hz, 1H), 6.09-6.03 (m, 1H), 3.03-2.93 (m, 1H), 2.73-2.65 (m, 2H), 2.33-2.27 (m, 2H);

(95) MS found (M+H).sup.+ (m/z), 303.8; calcd for C.sub.12H.sub.9N.sub.5O.sub.5 m/z, 303.06.

[14-4] Synthesis of 3-(8-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione

(96) To the solution of 3-(8-nitro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (0.02 g, 0.066 mmol) dissolved in AcOH/THF (2 ml/2 ml), iron (Fe, 0.02 g) was added, and it was stirred at a room temperature overnight. The reaction mixture was filtered and was extracted with ethyl acetate. The organic layer was dried Na.sub.2SO.sub.4, and was purified by column chromatography to obtain a pure compound of 0.004 g (22%) as a yellow-green solid.

(97) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.16 (s, 1H), 7.56 (t, J=7.9 Hz, 1H), 7.23-7.17 (m, 2H), 6.69 (s, 2H), 5.92-5.86 (m, 1H), 3.02-2.91 (m, 1H), 2.74-2.61 (m, 1H), 2.29-2.20 (m, 1H);

(98) MS found (M+H).sup.+ (m/z), 274.0; calcd for C.sub.12H.sub.11N.sub.5O.sub.3 m/z, 273.09.

Example 15

Synthesis of N-(3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-6-yl)acetamide (Chemical Formula 23)

(99) ##STR00031##

(100) To the solution of 3-(6-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (0.02 g, 0.07 mmol) dissolved in THF (5 ml), acetyl chloride (0.005 ml, 0.07 mmol) was added, and it was stirred at a room temperature overnight. The reaction mixture was extracted with ethyl acetate. The organic layer was dried on Na.sub.2SO.sub.4, and was purified by column chromatography to obtain a pure compound of 0.01 g (45%) as a white solid.

(101) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.18 (s, 1H), 10.71 (s, 1H), 8.61 (d, J=2.2 Hz, 1H), 8.23-8.13 (m, 2H), 5.98-5.93 (m, 1H), 3.02-2.90 (m, 1H), 2.70-2.63 (m, 2H), 2.30-2.23 (m, 1H), 2.15 (s, 3H);

(102) MS found (M+H).sup.+ (m/z), 315.9; calcd for C.sub.14H.sub.13N.sub.5O.sub.4 m/z, 315.10.

Example 16

Synthesis of 3-(8-amino-1-oxoisoquinolin-2(1H)-yl)piperidine-2,6-dione (Chemical Formula 31)

[16-1] Synthesis of Compound <16-1>

(103) ##STR00032##

(104) To the solution of 2-methyl-6-nitrobenzoic acid (15 g, 82.8 mmol) dissolved in acetone (350 ml), iodomethane (25.8 ml, 414.0 mmol) and potassium carbonate (57.2 g, 414.0 mmol) were added, and the reaction mixture was refluxed for 15 hours and was stirred. The reaction mixture was cooled to a room temperature and was filtered. The filtrates were concentrated, and the residues were diluted, and then were extracted with EtOAc (250 ml×2 times). The combined organic layer was dried on MgSO.sub.4, and the solvent was removed under vacuum to obtain yellow oil. The combined organic layer was concentrated under reduced pressure to obtain methyl 2-methyl-6-nitro benzoate (compound <16-1>) (15.9 g, 81.5 mmol, 98%) as a white solid.

[16-2] Synthesis of Compound <16-2>

(105) ##STR00033##

(106) To the solution of compound <16-1> (15.9 g, 81.5 mmol) dissolved in dichloroethane (150 ml), N-bromosuccinimide (16.9 g, 122.2 mmol) and benzoyl peroxide (197 mg, 0.815 mmol) were added at a room temperature, and the mixture was stirred under reflux for 8 hours. The reaction mixture was cooled to a room temperature and was filtered. The filtrates were concentrated. The residues were purified by silica-gel column chromatography using EA/Hx (8%) as an eluent, to obtain compound <16-2> (15.9 g, 58.0 mmol, 71%) as a lemon yellow.

(107) .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.98 (s, 3H), 4.57 (s, 2H), 7.59 (dd, 1H, J=7.8, 8.4 Hz), 7.78 (d, 1H, J=7.8 Hz), 8.1 (d, 1H, J=8.4 Hz).

[16-3] Synthesis of Compound <16-3>

(108) ##STR00034##

(109) To a stirred suspension of a fire-dried 4 Å molecular sieve dissolved in acetonitrile (150 ml), NMO (15.6 g, 133.4 mmol) was added. In 5 minutes, LDH-17-081 (15.9 g, 58.0 mmol) dissolved in acetonitrile (20 ml) was added. The reaction mixture was stirred at a room temperature for 1.5 hours, and it was filtered through silica-gel, and it was eluted with EtOAc, and it was concentrated under vacuum to obtain reddish brown oil. It was purified by silica-gel column chromatography (MPLC, 0 to 30% EA/Hx) to obtain compound <16-3> (8.12 g, 38.8 mmol, 67%) as a white solid.

(110) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 10.08 (s, 1H), 8.44 (dd, J=8.2, 1.2 Hz, 1H), 8.26 (dd, J=7.7, 1.2 Hz, 1H), 7.81 (t, J=8.0 Hz, 1H), 4.06 (s, 3H).

[16-4] Synthesis of Compound <16-4>

(111) ##STR00035##

(112) To the stirred suspension of (methoxymethyl)triphenyl phosphonium chloride (1.22 g, 3.59 mmol) dissolved in THF (24 ml), sodium hydride (192 mg, 4.78 mmol) was added in small portions in an ice container under N.sub.2. The red solution was stirred at a room temperature for 30 minutes, and then compound <16-3> (500 mg, 2.39 mmol) dissolved in THF (6 ml) was added dropwise. The reactants were stirred at a room temperature for 12 hours. The mixture was diluted with water, and was extracted with EtOAc (50 ml). The combined organic layer was washed with salt water, and was dried on MgSO.sub.4, and the solvent was removed under vacuum to obtain dark oil. The crude compound was purified by silica-gel column chromatography using EtOAc/Hex (30%) as an eluent, to obtain compound <16-4> (372 mg, 1.57 mmol, 66%) as yellow oil (mixture of 1.7:1 E/Z isomers).

[16-5] Synthesis of Compound <16-5>

(113) ##STR00036##

(114) To the solution of compound <16-4> (360 mg, 1.52 mmol) dissolved in MeOH (4 ml), H.sub.2O (4 ml) and THF (12 ml), lithium monohydrate (638 mg, 15.2 mmol) was added at a room temperature, and the reaction mixture was stirred at 70° C. overnight. The reaction mixture was concentrated to remove THF and methanol. Subsequently, water (40 ml) was added. The combined organic layer was concentrated under reduced pressure to obtain compound <16-5> (332 mg, 1.49 mmol, 98%) as a yellow solid (mixture of 1.7:1 E/Z isomers).

[16-6] Synthesis of Compound <16-7>

(115) ##STR00037##

(116) To the solution of compound <16-5> (332 mg, 1.49 mmol), 3-aminopiperidine-2,6-dione hydrochloride (1.08 g, 6.55 mmol), EDCl HCl (314 mg, 1.64 mmol), and HOBt H.sub.2O (221 mg, 1.64 mmol), dissolved in DMF (10 ml), DIPEA (1.71 ml, 9.82 mmol) was added at a room temperature, and the reaction mixture was stirred at a room temperature for 4 hours. The reaction mixture was diluted with water and was extracted with EtOAc (50 ml×2 times). The combined organic layer was dried on MgSO.sub.4, and the solvent was removed under vacuum to obtain yellow oil. The crude compound was purified by column chromatography using MeOH/DCM (5%) as an eluent, to obtain E-products (219 mg, 0.658 mmol, 44%), and Z-products (43 mg, 0.129 mmol, 9%) as lemon yellow solids.

(117) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.98-7.86 (m, 2H), 7.69 (d, J=7.9 Hz, 1H), 7.47 (t, J=8.1 Hz, 1H), 7.12 (d, J=12.8 Hz, 1H), 6.47 (d, J=6.5 Hz, 1H), 6.06 (d, J=12.8 Hz, 1H), 4.87-4.76 (m, 1H), 3.73 (s, 3H), 2.97-2.71 (m, 3H), 2.16-1.97 (m, 1H).

[16-7] Synthesis of Compound <16-8>

(118) ##STR00038##

(119) To the solution of compound <16-7> (100 mg, 0.300 mmol) dissolved in TFA (6 ml), the reaction mixture was stirred at 150° C. for 90 minutes under the monowave-assisted condition. Until the reactants were dried, it was evaporated. The residues were purified by silica-gel column chromatography using MeOH/DCM (5%) as an eluent to obtain compound <16-8> as an ivory solid.

(120) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.98 (s, 1H), 7.82-7.67 (m, 2H), 7.47 (d, J=7.3 Hz, 1H), 7.06 (d, J=7.5 Hz, 1H), 6.67 (d, J=7.4 Hz, 1H), 5.61-5.48 (m, 1H), 2.98-2.69 (m, 3H), 2.37-2.23 (m, 1H).

[16-8] Synthesis of Compound of Chemical Formula 31

(121) ##STR00039##

(122) To the solution of compound <16-8> (20 mg, 0.066 mmol) and Zn (63 mg, 0.957 mmol), dissolved in THF (0.5 ml), 2M aq. HCl (0.5 ml) was added, and the reaction mixture was stirred at a room temperature for 30 minutes. The mixture was filtered, and was distributed between ethyl acetate and saturated aqueous NaHCO.sub.3. The combined organic layer was dried on MgSO.sub.4, and the solvent was removed under vacuum to obtain compound of Chemical formula 31 (17 mg, 0.063 mmol, 94%) as a yellow solid.

(123) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 10.99 (s, 1H), 7.29 (t, J=7.8 Hz, 2H), 7.19 (d, J=7.4 Hz, 2H), 6.65-6.54 (m, 2H), 6.40 (d, J=7.5 Hz, 1H), 5.33 (s, 1H), 2.91-2.70 (m, 1H), 2.70-2.53 (m, 2H), 2.07-1.93 (m, 1H).

Example 17

Synthesis of 3-(5-amino-1-oxoisoquinolin-2(1H)-yl)piperidine-2,6-dione (Chemical Formula 24)

[17-1] Synthesis of 5-nitro-1H-isochromen-1-one

(124) The starting material (1 g, 5.1 mmol) was dissolved in DMF (8.2 ml) and DMFDMA (2.3 ml), and it was stirred at 115° C. for 17 hours. When the reaction was finished, it was concentrated to remove the solvent, and it was dissolved in EA, and then silica gel (80 g) was added, and it was stirred at a room temperature for 3 hours. The reactants were filtered, and were washed with EA, and then were concentrated, to obtain a desired compound, 5-nitro-1H-isochromen-1-one (800 mg, brown solid, 82%).

(125) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.62 (d, J=7.8 Hz, 1H), 8.47 (d, J=8.1 Hz, 1H), 7.65 (m, 1H), 7.42 (d, J=6.3 Hz, 1H), 7.36 (d, J=6.3 Hz, 1H).

[17-2] Synthesis of 3-(5-nitro-1-oxoisoquinolin-2(1H)-yl)piperidine-2,6-dione

(126) 5-nitro-1H-isochromen-1-one (1 g, 5.2 mmol) and 3-aminopiperidine-2,6-dione (0.67 g, 5.2 mmol) were dissolved in MeOH (10 ml), and then it was refluxed as heated for 5 hours, and then TEA (1.45 ml) was added, and the reaction was progressed overnight. When the reaction was completed, water was added to dilute it, and then it was extracted with EA and was concentrated under reduced pressure. The concentrates were dissolved in toluene, and then PPTS (0.1 g) was added and then was refluxed as heated. When the reaction was completed, it was neutralized with saturated NaHCO.sub.3 aqueous solution, and it was extracted with EA and was dried and was isolated and purified to obtain a desired compound, 3-(5-nitro-1-oxoisoquinolin-2(1H)-yl)piperidine-2,6-dione.

(127) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.78 (d, J=8.3 Hz, 1H), 8.48 (d, J=6.6 Hz, 1H), 8.03 (s, 1H), 7.65 (t, J=8.0 Hz, 1H), 7.44 (d, J=7.8 Hz, 1H), 7.17 (d, J=7.9 Hz, 1H), 5.45-5.37 (m, 1H), 3.04-2.57 (m, 3H), 2.48-2.17 (m, 1H).

[17-3] Synthesis of 3-(5-amino-1-oxoisoquinolin-2(1H)-yl)piperidine-2,6-dione

(128) 3-(5-Nitro-1-oxoisoquinolin-2(1H)-yl)piperidine-2,6-dione was dissolved in EA, and then ammonium formate and 10% Pd/C were added, and it was refluxed as heated. When the reaction was completed, it was filtered into Celite, and it was washed with EA, and then the filtrates were concentrated under reduced pressure to obtain a desired compound, 3-(5-amino-1-oxoisoquinolin-2(1H)-yl)piperidine-2,6-dione.

(129) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 9.68 (s, 1H), 7.90 (d, J=8.0 Hz, 1H), 7.40-7.27 (m, 1H), 7.19-6.87 (m, 2H), 6.53 (dd, J=17.0, 7.4 Hz, 1H), 5.42 (dd, J=42.5, 18.5 Hz, 1H), 4.27-3.86 (m, 2H), 3.07-2.14 (m, 4H);

(130) LC/MS (m/z) 272.1 (M+H).

Example 18

Synthesis of 3-(7-(aminomethyl)-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 32) and 1-(3-chloro-4-methylphenyl)-3-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-7-yl)methyl)urea (Chemical Formula 37)

(131) ##STR00040## ##STR00041##

[18-1] Synthesis of methyl 4-chloro-2-nitrobenzoate

(132) 4-chloro-2-nitrobenzoic acid (10 g, 50 mmol) was suspended in dry methanol (5 ml per 1 mmol), and it was cooled to 0° C. SOCl.sub.2 (22 ml, 250 mmol) was slowly added, and then the suspension was heated at 50° C. for 16 hours. The produced solution was concentrated under reduced pressure, and the residues were dissolved in water-saturated NaHCO.sub.3 solution (2.5 ml/1 mmol). The solution was extracted with ethyl acetate (2.5 ml×3 times/1 mmol). The combined organic layer was washed with saturated aqueous NaCl solution (7.5 ml per 1 mmol), and it was dried with Na.sub.2SO.sub.4 to filter it, and then the solution was concentrated under reduced pressure to obtain a pure compound of 9 g (70%) as an ivory solid.

(133) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.86 (d, J=1.8 Hz, 1H), 7.73 (d, J=8.3 Hz, 1H), 7.64 (dd, J=8.3, 1.9 Hz, 1H), 3.92 (s, 3H).

[18-2] Synthesis of methyl 4-(((tert-butoxy carbonyl)amino)methyl)-2-nitrobenzoate

(134) Potassium [[(tert-butoxy carbonyl)amino]methyl]trifluoroborate (2.6 g, 11 mmol), methyl 4-chloro-2-nitro benzoate (2 g, 9.28 mmol), Pd(OAc)2 (0.104 g, 0.464 mmol), SPhos ligand (0.38 g, 0.928 mmol) and K.sub.2CO.sub.3 (4 g, 28 mmol) were filled into a sealed tube. The mixture was washed with N.sub.2 3 times. subsequently, toluene/H.sub.2O (4:1, 20 ml/5 ml) was added to the reaction tube and it was deaerated with N.sub.2 for 30 minutes. The reaction mixture was stirred at 85° C. for 4 hours, and then it was cooled to a room temperature. The produced mixture was extracted with EtOAc, and then the organic layer was combined, and it was dried (MgSO.sub.4) and was filtered. The solvent was removed under vacuum and the products were purified by column chromatography to obtain a pure compound of 2.5 g (87%) as an ivory solid.

(135) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.79 (s, 1H), 7.72 (d, J=7.9 Hz, 1H), 7.58 (d, J=7.9 Hz, 1H), 5.04 (s, 1H), 4.41 (d, J=5.9 Hz, 2H), 3.91 (s, 3H), 1.46 (s, 9H).

[18-3] Synthesis of 4-(((tert-butoxy carbonyl)amino)methyl)-2-nitrobenzoic acid

(136) To the solution of 4-((tert-butoxy carbonyl)amino)methyl)-2-nitro benzoate (1.5 g, 4.8 mmol) dissolved in THF/MeOH (10 ml/10 ml), LiOH (0.480 g, 20 mmol) dissolved in H.sub.2O (2.5 ml) was added, and it was stirred at a room temperature for 2 hours. After the reaction was completed, the solvent was evaporated and it was extracted with H.sub.2O. HCl was added to the water layer, and it was extracted with EA. The organic layer was dried on Na.sub.2SO.sub.4 to obtain a pure compound of 1.4 g (98%) as a brown solid.

(137) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 13.81 (s, 1H), 7.82 (d, J=7.9 Hz, 1H), 7.78 (s, 1H), 7.62 (d, J=8.5 Hz, 1H), 7.58 (t, J=6.2 Hz, 1H), 4.24 (d, J=6.1 Hz, 2H), 1.39 (s, 9H).

[18-4] Synthesis of tert-butyl (4-((2,6-dioxopiperidin-3-yl)carbamoyl)-3-nitrobenzyl)carbamate

(138) To 4-((tert-butoxy carbonyl)amino)methyl)-2-nitrobenzoic acid (1.4 g, 3.85 mmol), EDCl-HCl (0.813 mg, 4.24 mmol) and HOBt (0.649 mg, 4.24 mmol) were added, and it was stirred at a room temperature for 30 minutes. To the reaction mixture, 3-aminopiperidine-2,6-dione hydrochloride (1.3 g, 7.7 mmol) and DIPEA (2.7 ml, 15.4 mmol) were added, and it was stirred at a room temperature for 16 hours. After the reaction was finished, the mixture was extracted with EA. The organic layer was dried on Na.sub.2SO.sub.4, and was purified by column chromatography to obtain a pure compound 1.05 g (67%) as a white-green solid.

(139) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.88 (s, 1H), 9.01 (d, J=8.3 Hz, 1H), 7.89 (s, 1H), 7.67 (d, J=8.0 Hz, 1H), 7.61-7.58 (m, 2H), 4.75-4.70 (m, 1H), 4.24 (d, J=6.1 Hz, 2H), 2.82-2.75 (m, 1H), 2.57-2.49 (m, 1H), 2.06-1.99 (m, 2H).

[18-5] Synthesis of tert-butyl (3-amino-4-((2,6-dioxopiperidin-3-yl)carbamoyl)benzyl)carbamate

(140) To tert-butyl(4-((2,6-dioxopiperidin-3-yl)carbamoyl)-3-nitrobenzyl)carbamate (1 g, 2.5 mmol) dissolved in MeOH/EA (50 ml/50 ml), 1 Pd/C (0.1 g) was added, and the reaction mixture was stirred under the hydrogen gas circumstance at a room temperature for 16 hours. The solution was filtered through Celite, and it was dried under vacuum to obtain a pure compound of 932 mg (99%) as a grey-white solid.

(141) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 10.83 (s, 1H), 8.40 (d, J=8.3 Hz, 1H), 7.46 (d, J=8.2 Hz, 1H), 7.32 (t, J=6.3 Hz, 1H), 6.55 (d, J=1.6 Hz, 1H), 6.49 (s, 2H), 6.40 (d, J=8.1 Hz, 1H), 4.76-4.67 (m, 1H), 4.00 (d, J=6.3 Hz, 2H), 2.84-2.72 (m, 1H), 2.56-2.54 (m, 1H), 2.16-2.03 (m, 1H), 1.99-1.90 (m, 1H).

[18-6] Synthesis of tert-butyl ((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-7-yl)methyl)carbamate

(142) To the solution of tert-butyl(3-amino-4-((2,6-deoxypiperidin-3-yl)carbamoyl)benzyl)carbamate (0.922 g, 2.45 mmol) dissolved in glacial acetic acid (10 ml), sodium nitrite (0.288 g, 4.17 mmol) was added, and it was stirred at a room temperature for 1 hour. The reaction mixture was extracted with EA. The organic layer was dried on Na.sub.2SO.sub.4, and was purified by column chromatography to obtain a pure compound of 0.613 g (65%) as a white solid.

(143) .sup.1H NMR (300 MHz, Chloroform-d) δ 8.32 (d, J=8.2 Hz, 1H), 8.09 (s, 1H), 8.02 (s, 1H), 7.77 (d, J=8.2 Hz, 1H), 5.86-5.80 (m, 1H), 5.11 (s, 1H), 4.56 (d, J=6.3 Hz, 2H), 3.04-2.83 (m, 3H), 2.44-2.37 (m, 1H), 1.48 (s, 9H).

[18-7] Synthesis of 3-(7-(aminomethyl)-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione hydrochloride

(144) To the solution of tert-butyl((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-7-yl)methyl)carbamate (0.605 mg, 1.56 mmol) dissolved in DCM (10 ml), 4M HCl mixed to 1,4-dioxane (2 ml) was added, and it was stirred at a room temperature for 16 hours. the solvent was dried under vacuum to obtain a compound of 0.545 g (quant.) (Chemical formula 32) as an ivory solid.

(145) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 11.22 (s, 1H), 8.65 (s, 3H), 8.40 (d, J=1.5 Hz, 1H), 8.33 (d, J=8.1 Hz, 1H), 8.06 (dd, J=8.2, 1.6 Hz, 1H), 6.02-5.99 (m, 1H), 4.35 (q, J=5.7 Hz, 2H), 3.02-2.94 (m, 1H), 2.75-2.66 (m, 2H), 2.32-2.27 (m, 1H).

[18-8] Synthesis of 1-(3-chloro-4-methylphenyl)-3-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-7-yl)methyl)urea (Chemical Formula 37)

(146) The mixture of 3-(7-(aminomethyl)-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione hydrochloride (0.2 g, 0.62 mmol), 3-chloro-4-methylphenyl isocyanate (0.104 g, 0.62 mmol) and TEA (0.2 ml, 1.24 mmol), dissolved in THF (10 ml) was heated from a room temperature to 40° C. for 2 hours. The mixture was extracted with EA and was purified by column chromatography to obtain the pure compound of Chemical formula 37 of 0.134 g (48%) as an ivory solid.

(147) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 11.19 (s, 1H), 8.92 (s, 1H), 8.25 (d, J=8.2 Hz, 1H), 8.11 (s, 1H), 7.90 (d, J=8.1 Hz, 1H), 7.66 (d, J=2.0 Hz, 1H), 7.20-7.14 (m, 2H), 7.00 (t, J=6.1 Hz, 1H), 6.00-5.96 (m, 1H), 4.56 (d, J=6.0 Hz, 2H), 3.00-2.93 (m, 1H), 2.73-2.66 (m, 2H), 2.30-2.25 (m, 1H), 2.23 (s, 3H).

Example 19

Synthesis of 1-(3-chloro-4-methylphenyl)-3-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-5-yl)methyl)urea (Chemical Formula 39)

(148) The compound of Chemical formula 39 was prepared according to the preparation method of Example 18.

(149) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.21 (s, 1H), 8.97 (s, 1H), 8.17-8.07 (m, 2H), 7.88 (d, J=7.1 Hz, 1H), 7.64 (d, J=2.1 Hz, 1H), 7.17 (d, J=8.3 Hz, 1H), 7.09 (dd, J=8.2, 2.2 Hz, 1H), 6.81 (t, J=6.3 Hz, 1H), 6.03-5.98 (m, 1H), 4.84 (d, J=6.1 Hz, 2H), 3.04-2.93 (m, 1H), 2.79-2.65 (m, 2H), 2.34-2.25 (m, 1H), 2.22 (s, 3H).

Example 20

Synthesis of 1-(3-chlorophenyl)-3-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-7-yl)methyl)urea (Chemical Formula 40)

(150) The compound of Chemical formula 40 was prepared according to the preparation method of Example 18.

(151) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.17 (s, 1H), 9.01 (s, 1H), 8.26 (d, J=8.2 Hz, 1H), 8.11 (s, 1H), 7.91 (dd, J=8.2, 1.6 Hz, 1H), 7.68 (s, 1H), 7.25-7.23 (m, 2H), 7.02 (t, J=6.1 Hz, 1H), 6.98-6.93 (m, 1H), 6.01-5.95 (m, 1H), 4.57 (d, J=6.0 Hz, 2H), 3.03-2.92 (m, 1H), 2.77-2.64 (m, 2H), 2.31-2.24 (m, 1H).

Example 21

Synthesis of 1-(4-chlorophenyl)-3-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-7-yl)methyl)urea (Chemical Formula 41)

(152) The compound of Chemical formula 41 was prepared according to the preparation method of Example 18.

(153) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.18 (s, 1H), 8.93 (s, 1H), 8.25 (d, J=8.2 Hz, 1H), 8.11 (s, 1H), 7.90 (dd, J=8.2, 1.6 Hz, 1H), 7.48-7.43 (m, 2H), 7.29-7.24 (m, 2H), 6.97 (t, J=6.0 Hz, 1H), 6.01-5.95 (m, 1H), 4.57 (d, J=5.9 Hz, 2H), 3.03-2.92 (m, 1H), 2.77-2.63 (m, 2H), 2.31-2.24 (m, 1H).

Example 22

Synthesis of 1-(3,4-dichlorophenyl)-3-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-7-yl)methyl)urea (Chemical Formula 42)

(154) The compound of Chemical formula 42 was prepared according to the preparation method of Example 18.

(155) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.18 (s, 1H), 9.13 (s, 1H), 8.26 (d, J=8.2 Hz, 1H), 8.11 (s, 1H), 7.90 (dd, J=8.2, 1.6 Hz, 1H), 7.85 (d, J=2.5 Hz, 1H), 7.46 (d, J=8.8 Hz, 1H), 7.30 (dd, J=8.9, 2.5 Hz, 1H), 7.09 (t, J=6.1 Hz, 1H), 6.01-5.95 (m, 1H), 4.57 (d, J=6.0 Hz, 2H), 3.03-2.91 (m, 1H), 2.77-2.64 (m, 2H), 2.31-2.23 (m, 1H).

Example 23

Synthesis of 1-(4-chloro-3-(trifluoromethyl)phenyl)-3-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-7-yl)methyl)urea (Chemical Formula 43)

(156) The compound of Chemical formula 43 was prepared according to the preparation method of Example 18.

(157) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.17 (s, 1H), 9.34 (s, 1H), 8.26 (d, J=8.2 Hz, 1H), 8.12 (s, 1H), 8.07 (d, J=2.5 Hz, 1H), 7.91 (dd, J=8.2, 1.6 Hz, 1H), 7.63 (dd, J=8.8, 2.5 Hz, 1H), 7.55 (d, J=8.7 Hz, 1H), 7.17 (t, J=6.2 Hz, 1H), 6.01-5.95 (m, 1H), 4.58 (d, J=5.9 Hz, 2H), 3.03-2.90 (m, 1H), 2.76-2.63 (m, 2H), 2.31-2.23 (m, 1H).

Example 24

Synthesis of 1-(3-chloro-4-methoxyphenyl)-3-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-7-yl)methyl)urea (Chemical Formula 44)

(158) The compound of Chemical formula 44 was prepared according to the preparation method of Example 18.

(159) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.17 (s, 1H), 8.78 (s, 1H), 8.25 (d, J=8.2 Hz, 1H), 8.11 (s, 1H), 7.90 (dd, J=8.1, 1.6 Hz, 1H), 7.63 (d, J=2.6 Hz, 1H), 7.23 (dd, J=8.9, 2.6 Hz, 1H), 7.03 (d, J=9.0 Hz, 1H), 6.95 (t, J=6.0 Hz, 1H), 6.01-5.95 (m, 1H), 4.56 (d, J=6.0 Hz, 2H), 3.03-2.91 (m, 1H), 2.77-2.63 (m, 2H), 2.32-2.24 (m, 1H).

Example 25

Synthesis of Other Compounds

[25-1] Synthesis of 3-chloro-N-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-6-yl)methyl)-4-methylbenzamide (Chemical Formula

(160) The compound of Chemical formula 46 was prepared according to the preparation method of Example 18.

(161) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.20 (s, 1H), 9.35 (t, J=5.9 Hz, 1H), 8.26 (d, J=8.1 Hz, 1H), 8.14 (s, 1H), 7.98 (d, J=1.7 Hz, 1H), 7.93 (d, J=8.2 Hz, 1H), 7.81 (dd, J=8.0, 1.8 Hz, 1H), 7.50 (d, J=7.9 Hz, 1H), 5.99 (dd, J=12.3, 5.4 Hz, 1H), 4.74 (d, J=5.5 Hz, 2H), 3.07-2.88 (m, 1H), 2.75-2.63 (m, 2H), 2.40 (s, 3H), 2.35-2.22 (m, 1H).

[25-2] Synthesis of 3-(6-(aminomethyl)-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 33)

(162) The compound of Chemical formula 33 was prepared according to the preparation method of Example 14.

(163) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 11.22 (s, 1H), 8.57 (s, 3H), 8.44 (d, J=1.9 Hz, 1H), 8.33 (d, J=8.4 Hz, 1H), 8.22 (dd, J=8.5, 1.9 Hz, 1H), 6.05-6.00 (m, 1H), 4.32 (q, J=5.9 Hz, 2H), 3.02-2.94 (m, 1H), 2.76-2.67 (m, 2H), 2.32-2.28 (m, 1H).

[25-3] Synthesis of 3-(5-(aminomethyl)-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 34)

(164) The compound of Chemical formula 34 was prepared according to the preparation method of Example 14.

(165) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 11.24 (s, 1H), 8.36 (s, 3H), 8.32 (d, J=8.1 Hz, 1H), 8.19 (t, J=7.8 Hz, 1H), 7.99 (d, J=7.5 Hz, 1H), 6.04-6.01 (m, 1H), 4.66-4.55 (m, 2H), 3.01-2.93 (m, 1H), 2.73-2.63 (m, 2H), 2.29-2.23 (m, 1H).

[25-4] Synthesis of 1-(3-chlorophenyl)-3-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-7-yl)methyl)thiourea (Chemical Formula 52)

(166) The compound of Chemical formula 52 was prepared according to the preparation method of Example 18.

(167) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 11.20 (s, 1H), 10.00 (s, 1H), 8.60 (s, 1H), 8.26 (d, J=8.2 Hz, 1H), 8.12 (s, 1H), 7.92 (dd, J=8.2, 1.6 Hz, 1H), 7.68 (s, 1H), 7.36 (d, J=5.1 Hz, 2H), 7.19-7.17 (m, 1H), 6.00-5.97 (m, 1H), 5.01 (d, J=5.8 Hz, 2H), 3.00-2.93 (m, 1H), 2.74-2.63 (m, 2H), 2.30-2.27 (m, 1H).

[25-5] Synthesis of 2-(3-chlorophenyl)-N-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-7-yl)methyl)acetamide (Chemical Formula 53)

(168) The compound of Chemical formula 53 was prepared according to the preparation method of Example 18.

(169) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 11.20 (s, 1H), 8.85 (t, J=6.0 Hz, 1H), 8.22 (d, J=8.1 Hz, 1H), 8.04 (s, 1H), 7.83 (dd, J=8.3, 1.3 Hz, 1H), 7.36-7.30 (m, 3H), 7.26 (d, J=7.4 Hz, 1H), 5.99-5.96 (m, 1H), 4.55 (d, J=6.0 Hz, 2H), 3.58 (s, 2H), 3.00-2.92 (m, 1H), 2.73-2.64 (m, 2H), 2.29-2.25 (m, 1H).

[25-6] Synthesis of 3-(4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 25)

(170) The compound of Chemical formula 25 was prepared according to the preparation method of Example 14.

(171) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.40 (d, J=7.1 Hz, 1H), 8.23 (d, J=8.2 Hz, 1H), 8.09 (s, 1H), 8.03 (t, J=7.7 Hz, 1H), 7.88 (t, J=7.6 Hz, 1H), 5.89-5.83 (m, 1H), 3.08-2.83 (m, 3H), 2.52-2.31 (m, 1H).

[25-7] Synthesis of 3-(7-nitro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 26)

(172) The compound of Chemical formula 26 was prepared according to the preparation method of Example 14.

(173) .sup.1H NMR (300 MHz, Methanol-d.sub.4) δ 9.03 (s, 1H), 8.70 (d, J=8.7 Hz, 1H), 8.58 (d, J=8.8 Hz, 1H), 6.14-5.99 (m, 1H), 3.12-2.85 (m, 3H), 2.54-2.38 (m, 1H).

[25-8] Synthesis of 3-(6-nitro-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 27)

(174) The compound of Chemical formula 27 was prepared according to the preparation method of Example 14.

(175) .sup.1H NMR (300 MHz, Methanol-d.sub.4) δ 9.11 (d, J=2.5 Hz, 1H), 8.85 (d, J=11.5 Hz, 1H), 8.46 (d, J=8.7 Hz, 1H), 6.29-5.87 (m, 1H), 3.33-2.71 (m, 3H), 2.52-2.31 (m, 1H).

[25-9] Synthesis of 3-(7-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 28)

(176) The compound of Chemical formula 28 was prepared according to the preparation method of Example 14.

(177) .sup.1H NMR (500 MHz, Methanol-d.sub.4) δ 7.99 (d, J=8.3 Hz, 1H), 7.20-7.10 (m, 2H), 5.89 (dd, J=12.2, 5.4 Hz, 1H), 3.02-2.79 (m, 3H), 2.39-2.32 (m, 1H).

[25-10] Synthesis of 3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-5-carboxylic acid (Chemical Formula 29)

(178) The compound of Chemical formula 29 was prepared according to the preparation method of Example 14.

(179) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 13.50 (s, 1H), 11.23 (s, 1H), 8.33 (dd, J=8.2, 1.1 Hz, 1H), 8.17 (t, J=7.8 Hz, 1H), 7.94 (dd, J=7.4, 1.1 Hz, 1H), 6.03-5.97 (m, 1H), 3.03-2.90 (m, 1H), 2.75-2.60 (m, 2H), 2.34-2.25 (m, 1H).

[25-11] Synthesis of 3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)-3-methylpiperidine-2,6-dione (Chemical Formula 30)

(180) The compound of Chemical formula 30 was prepared according to the preparation method of Example 14.

(181) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 11.03 (s, 1H), 7.68 (t, J=8.0 Hz, 1H), 7.18-7.17 (m, 3H), 6.97 (d, J=7.9 Hz, 1H), 2.96-2.88 (m, 1H), 2.73-2.67 (m, 1H), 2.63-2.58 (m, 1H), 2.01-1.97 (m, 1H), 1.95 (s, 3H).

[25-12] Synthesis of 1-(3-chloro-4-methylphenyl)-3-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-6-yl)methyl)urea (Chemical Formula 38)

(182) The compound of Chemical formula 38 was prepared according to the preparation method of Example 18.

(183) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.19 (s, 1H), 8.90 (s, 1H), 8.23 (d, J=8.3 Hz, 1H), 8.17 (s, 1H), 8.05 (d, J=8.5 Hz, 1H), 7.65 (s, 1H), 7.20-7.13 (m, 2H), 6.98 (t, J=6.1 Hz, 1H), 6.01-5.95 (m, 1H), 4.52 (s, 2H), 3.03-2.90 (m, 1H), 2.76-2.63 (m, 2H), 2.32-2.23 (m, 1H), 2.23 (s, 3H).

[25-13] Synthesis of 3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-5-carboxamide (Chemical Formula 54)

(184) The compound of Chemical formula 54 was prepared according to the preparation method of Example 14.

(185) .sup.1H NMR (500 MHz, Chloroform-d) δ 8.29 (d, J=8.1 Hz, 1H), 8.16 (s, 1H), 8.04 (t, J=7.8 Hz, 1H), 7.95 (d, J=7.4 Hz, 1H), 6.24 (s, 1H), 5.96 (s, 1H), 5.82-5.79 (m, 1H), 3.02-2.93 (m, 2H), 2.87-2.79 (m, 1H), 2.42-2.37 (m, 1H).

[25-14] Synthesis of 3-(2,6-dioxopiperidin-3-yl)-N-methyl-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-5-carboxamide (Chemical Formula 55)

(186) The compound of Chemical formula 55 was prepared according to the preparation method of Example 14.

(187) .sup.1H NMR (500 MHz, Chloroform-d) δ 8.26 (d, J=8.0 Hz, 1H), 8.13 (s, 1H), 8.01 (t, J=7.8 Hz, 1H), 7.86 (d, J=7.3 Hz, 1H), 6.11 (d, J=5.2 Hz, 1H), 5.84-5.81 (m, 1H), 3.07 (d, J=4.9 Hz, 3H), 3.00-2.91 (m, 2H), 2.84-2.77 (m, 1H), 2.39-2.34 (m, 1H).

[25-15] Synthesis of N-(3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-5-yl)benzamide (Chemical Formula 56)

(188) The compound of Chemical formula 56 was prepared according to the preparation method of Example 14.

(189) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 12.49 (s, 1H), 11.26 (s, 1H), 9.12 (d, J=8.0 Hz, 1H), 8.20 (t, J=8.2 Hz, 1H), 8.02-7.99 (m, 3H), 7.73-7.61 (m, 3H), 6.17-6.11 (m, 1H), 3.02-2.94 (m, 1H), 2.77-2.67 (m, 2H), 2.37-2.30 (m, 1H).

Example 26

Synthesis of 3-chloro-N-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-6-yl)methyl)-4-methylbenzamide (Chemical Formula 45)

(190) ##STR00042##

(191) To 3-chloro-4-methylbenzoic acid (16 mg, 0.093 mmol), EDCl-HCl (20 mg, 0.102 mmol) and HOBt (16 mg, 0.102 mmol) were added, and it was stirred at a room temperature for 30 minutes. To the reaction mixture, 3-(6-(aminomethyl)-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione hydrochloride (30 mg, 0.093 mmol) and DIPEA (0.03 ml, 0.186 mmol) were treated, and it was stirred at a room temperature for 16 hours. After the reaction was completed, the mixture was extracted with EA. The organic layer was dried on Na.sub.2SO.sub.4, and was purified by column chromatography to obtain a pure compound of 24 mg (60%) as a white solid.

(192) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.19 (s, 1H), 9.34 (t, J=5.9 Hz, 1H), 8.24 (d, J=8.3 Hz, 1H), 8.18 (s, 1H), 8.08 (d, J=8.2 Hz, 1H), 7.95 (s, 1H), 7.79 (d, J=8.0 Hz, 1H), 7.49 (d, J=8.0 Hz, 1H), 6.00-5.95 (m, 1H), 4.70 (d, J=5.9 Hz, 2H), 3.01-2.86 (m, 1H), 2.76-2.63 (m, 2H), 2.39 (s, 3H), 2.30-2.23 (m, 1H).

Example 27

Synthesis of 1-(3-chloro-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydroisoquinolin-7-yl)methyl)urea (Chemical Formula 47)

(193) ##STR00043## ##STR00044##

[27-1] Synthesis of methyl 5-bromo-2-methylbenzoate (Compound <27-2>)

(194) To the solution of compound <27-1> (5 g, 23.3 mmol) dissolved in DMF (60 ml), iodomethane (7.24 ml, 116.3 mmol) and potassium carbonate (16.1 g, 116.3 mmol) were added at a room temperature and the reaction mixture was stirred at a room temperature for 4 hours. The reaction mixture was cooled to a room temperature and was filtered. The filtrates were concentrated. The residues were diluted with water, and it was extracted with EtOAc (250 ml×2 times). The combined organic layer was dried on MgSO.sub.4, and the solvent was removed under vacuum to obtain compound <27-2> (methyl 5-bromo-2-methylbenzoate) (5.34 g, 23.3 mmol, 99%) as yellow oil.

(195) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.78 (d, J=8.3 Hz, 1H), 7.49-7.31 (m, 2H), 3.88 (s, 3H), 2.58 (s, 3H).

[27-2] Synthesis of methyl 5-bromo-2-(bromomethyl)benzoate (Compound <27-3>)

(196) To the solution of compound <27-2> (2 g, 8.74 mmol) dissolved in dichloroethane (45 ml), N-bromosuccinimide (1.81 g, 13.12 mmol) and benzoyl peroxide (22 mg, 0.087 mmol) were added at a room temperature, and it was refluxed for 7 hours. The reaction mixture was cooled to a room temperature. The residues were purified by silica-gel column chromatography using EA/Hx (8%) as an eluent to obtain compound <27-3> (methyl 5-bromo-2-(bromomethyl)benzoate) (2.06 g, 6.69 mmol, 77%) as a white solid.

(197) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.84 (d, J=8.4 Hz, 1H), 7.63 (d, J=1.9 Hz, 1H), 7.51 (dd, J=8.4, 2.0 Hz, 1H), 4.90 (s, 2H), 3.94 (s, 3H).

[27-3] methyl 5-bromo-2-formylbenzoate (Compound <27-4>)

(198) To the stirred suspension of an oven-dried 4 Å molecular sieve in acetonitrile (30 ml), NMO (1.80 g, 15.4 mmol) was added. In 5 minutes, compound 3 (2.06 g, 6.69 mmol) dissolved in acetonitrile (10 ml) was added. The reaction mixture was stirred at a room temperature for 1.5 hours, and it was filtered through silica-gel, and it was eluted with EtOAc and it was concentrated under vacuum to obtain reddish brown oil. The residues were purified by silica-gel column chromatography using EA/Hx (10%) as an eluent to obtain compound <27-4> (methyl 5-bromo-2-formyl benzoate) (977 mg, 4.02 mmol, 60%) as a white solid.

(199) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 10.61 (s, 1H), 8.06 (d, J=2.1 Hz, 1H), 7.88 (d, J=8.3 Hz, 1H), 7.77 (dd, J=8.3, 2.1 Hz, 1H), 3.98 (s, 3H).

[27-4] Synthesis of methyl (E)-5-bromo-2-(2-methoxyvinyl)benzoate (Compound <27-5>)

(200) ##STR00045##

(201) To the stirred suspension of (methoxymethyl)triphenylphosphonium chloride (1.06 g, 3.09 mmol) dissolved in THF (24 ml) under N.sub.2 in an ice container, sodium hydride (165 mg, 4.12 mmol) was added in small portions. The red solution was stirred at a room temperature for 30 minutes, and then compound <27-4> (500 mg, 2.06 mmol) dissolved in THF (6 ml) dropwise. The reactants were stirred at a room temperature for 2 hours. The mixture was diluted with water and it was extracted with EtOAc (50 ml). The combined organic layer was washed with salt water, and it was dried on MgSO.sub.4, and the solvent was removed under vacuum to obtain dark oil. The crude compound was purified by silica-gel column chromatography using EtOAc/Hex (30%) as an eluent to obtain compound <27-5> (methyl (E)-5-bromo-2-(2-methoxyvinyl)benzoate) (348 mg, 1.28 mmol, 61%) as yellow oil (mixture of about 5:1 E/Z isomers).

(202) Compound <27-5-1>: .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.76 (d, J=8.5 Hz, 1H), 7.56 (d, J=2.0 Hz, 1H), 7.32 (dd, J=8.5, 2.0 Hz, 1H), 7.00 (d, J=12.9 Hz, 1H), 6.72 (d, J=12.9 Hz, 1H), 3.88 (s, 3H), 3.73 (s, 3H).

(203) Compound <27-5-2>: .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.25 (d, J=2.0 Hz, 1H), 7.69 (d, J=8.4 Hz, 1H), 7.30 (dd, J=8.4, 2.0 Hz, 1H), 6.26 (d, J=7.3 Hz, 1H), 6.07 (d, J=7.3 Hz, 1H), 3.87 (s, 3H), 3.80 (s, 3H).

[27-5] Synthesis of methyl (E)-5-(((tert-butoxy carbonyl)amino)methyl)-2-(2-methoxyvinyl)benzoate (Compound <27-6>)

(204) (Tert-butoxy carbonyl)amino)methyl)trifluoro borate (105 mg, 0.443 mmol), compound <27-5> (300 mg, 1.11 mmol), Pd(OAc)2 (12 mg, 0.055 mmol), SPhos (45 mg, 0.111 mmol) and potassium carbonate (459 mg, 3.32 mmol) were filled to a sealed tube. Then, toluene/H.sub.2O was added to the reaction tube, and it was deaerated with N.sub.2 for 30 minutes. The reaction mixture was stirred at 85° C. overnight, and then it was cooled to a room temperature. The produced mixture was extracted with EtOAc, and the organic layer was combined and it was dried with MgSO.sub.4, and it was filtered. The solvent was removed under vacuum, and the mixture was purified by silica-gel column chromatography (MPLC) using EA/Hx (10%) as an eluent to obtain compound <27-6> (methyl (E)-5-(((tert-butoxy carbonyl)amino)methyl)-2-(2-methoxyvinyl)benzoate) (233 mg, 0.725 mmol, 65%) as colorless oil.

(205) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.86 (d, J=8.1 Hz, 1H), 7.29 (d, J=1.7 Hz, 1H), 7.10 (dd, J=8.1, 1.7 Hz, 1H), 6.98 (d, J=12.9 Hz, 1H), 6.75 (d, J=12.9 Hz, 1H), 4.93 (s, 1H), 4.31 (d, J=6.1 Hz, 2H), 3.88 (s, 3H), 3.72 (s, 3H), 1.46 (s, 9H);

(206) LC/MS (ESI) m/z [M+H]+: 322.0.

[27-6] Synthesis of (E)-5-(((tert-butoxy carbonyl)amino)methyl)-2-(2-methoxyvinyl)benzoic acid (Compound <27-7>)

(207) To the solution of compound <27-6> (233 mg, 0.725 mmol) dissolved in MeOH (1.5 ml), H.sub.2O (1.5 ml) and THF (4.5 ml), lithium hydroxide monohydrate (152 mg, 3.63 mmol) was added at a room temperature and it was stirred at 50° C. overnight. The reaction mixture was concentrated to remove THF and methanol. Subsequently, water (50 mL) was added, and 1N HCl (aqueous) was used to adjust it to pH 3, and it was extracted with EtOAc (50 ml×2 times). The combined organic layer was concentrated under reduced pressure to obtain <27-7> ((E)-5-(((tert-butoxy carbonyl)amino)methyl)-2-(2-methoxyvinyl)benzoic acid) (232 mg, 0.725 mmol, quant.) as yellow gel.

(208) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.99 (d, J=8.3 Hz, 1H), 7.31 (s, 1H), 7.14 (d, J=8.0 Hz, 1H), 6.99 (d, J=12.8 Hz, 1H), 6.81 (d, J=12.9 Hz, 1H), 5.00-4.83 (m, 1H), 4.33 (s, 2H), 3.73 (s, 3H), 1.47 (s, 9H);

(209) LC/MS (ESI) m/z [M−H].sup.−: 306.0.

[27-7] Synthesis of tert-butyl (E)-(3-((2,6-dioxopiperidin-3-yl)carbamoyl)-4-(2-methoxyvinyl)benzyl)carbamate (Compound <27-8>)

(210) To the solution of compound <27-7> (223 mg, 0.798 mmol), 3-aminopiperidine-2,6-dione hydrochloride (239 mg, 1.45 mmol), EDCl HCl (153 mg, 0.798 mmol), and HOBt H.sub.2O (108 mg, 0.798 mmol), dissolved in DMF (10 ml), DIPEA (0.505 ml, 2.90 mmol) was added at a room temperature, and the reaction mixture was stirred at a room temperature for 3 hours. The reaction mixture was diluted with water, and it was extracted with EtOAc (50 ml×2 times). the combined organic layer was dried on MgSO.sub.4, and the solvent was removed under vacuum to obtain yellow oil. The crude compound was purified by silica-gel column chromatography using MeOH/DCM (5%) as an eluent to obtain compound <27-8> (tert-butyl (E)-(3-((2,6-dioxopiperidin-3-yl)carbamoyl)-4-(2-methoxyvinyl)benzyl)carbamate) (124 mg, 0.297 mmol, 41%) as a sky-blue solid.

(211) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 10.85 (s, 1H), 8.51 (d, J=8.4 Hz, 1H), 7.47-7.36 (m, 2H), 7.28 (d, J=7.7 Hz, 1H), 7.17 (d, J=12.9 Hz, 1H), 7.05 (d, J=7.9 Hz, 1H), 6.28 (d, J=12.8 Hz, 1H), 4.78-4.68 (m, 1H), 4.11 (d, J=6.3 Hz, 2H), 3.65 (s, 3H), 2.81-2.72 (m, 1H), 2.14-1.92 (m, 3H), 1.40 (s, 9H);

(212) LC/MS (ESI) m/z [M+H]+: 417.9, [M−H]−: 416.0.

[27-8] Synthesis of 3-(7-(aminomethyl)-1-oxoisoquinolin-2(1H)-yl)piperidine-2,6-dione hydrochloride (Compound <27-9>)

(213) To the solution of compound <27-8> (30 mg, 0.072 mmol) dissolved in 1,4-dioxane (1 ml), 4N HCl mixed with 1,4-dioxane (0.180 ml, 0.720 mmol) was added, and it was stirred at a room temperature for 2 hours. Then, the reaction mixture was stirred under the monowave-assisted condition at 150° C. for 1 hour. The reactants were evaporated. The residues were purified by silica-gel column chromatography using MeOH/DCM (5%) as an eluent to obtain compound <27-9> (3-(7-(aminomethyl)-1-oxoisoquinolin-2(1H)-yl)piperidine-2,6-dione hydrochloride) (26 mg, 0.072 mmol, quant.) as an ivory solid.

(214) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.06 (s, 1H), 8.40 (s, 3H), 8.24 (d, J=8.2 Hz, 1H), 7.74 (s, 1H), 7.61 (d, J=8.3 Hz, 1H), 7.48 (d, J=7.3 Hz, 1H), 6.67 (d, J=7.4 Hz, 1H), 5.62-5.43 (m, 1H), 4.20 (s, 2H), 2.95-2.79 (m, 1H), 2.77-2.58 (m, 2H), 2.13-1.98 (m, 1H);

(215) LC/MS (ESI) m/z [M+H]+: 286.9, [M−H]−: 284.9.

[27-9] Synthesis of 1-(3-chloro-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydroisoquinolin-7-yl)methyl)urea (Compound <27-10>)

(216) To the solution of compound <27-9> (23 mg, 0.071 mmol) dissolved in THF (0.5 ml), 3-chloro-4-methylphenyl isocyanate (12 mg, 0.071 mmol) and TEA (0.020 ml, 0.142 mmol) were added at a room temperature, and it was stirred at 40° C. overnight. The solvent was removed under vacuum, and the mixture was purified by silica-gel column chromatography (MPLC) using MeOH/DCM (6%) as an eluent to obtain <Chemical formula 47> (1-(3-chloro-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydroisoquinolin-7-yl)methyl)urea) (18 mg, 0.040 mmol, 56%) as a white solid.

(217) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.06 (s, 1H), 8.81 (s, 1H), 8.17 (d, J=8.3 Hz, 1H), 7.68 (s, 1H), 7.55 (s, 1H), 7.49-7.38 (m, 2H), 7.22-7.12 (m, 2H), 6.84 (t, J=5.8 Hz, 1H), 6.67 (d, J=7.4 Hz, 1H), 5.62-5.39 (m, 1H), 4.44 (d, J=5.8 Hz, 2H), 2.90-2.78 (m, 1H), 2.67-2.57 (m, 2H), 2.24 (s, 3H), 2.10-1.98 (m, 1H);

(218) LC/MS (ESI) m/z [M+H]+: 454.9, [M−H]−: 452.8.

Example 28

Synthesis of 1-(4-chloro-3-(trifluoromethyl)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydroisoquinolin-7-yl)methyl)urea (Chemical Formula 48)

(219) The compound of Chemical formula 48 was prepared according to the preparation method of Example 27.

(220) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.05 (s, 1H), 9.26 (s, 1H), 8.17 (d, J=8.3 Hz, 1H), 8.10 (s, 1H), 7.63 (d, J=9.1 Hz, 1H), 7.56 (d, J=6.4 Hz, 2H), 7.44 (dd, J=15.5, 7.9 Hz, 2H), 7.04 (s, 1H), 6.67 (d, J=7.4 Hz, 1H), 5.60-5.42 (m, 1H), 4.46 (d, J=5.9 Hz, 2H), 2.88-2.78 (m, 1H), 2.74-2.59 (m, 2H), 2.12-1.94 (m, 1H).

Example 29

Synthesis of 1-(3-chloro-4-methoxyphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydroisoquinolin-7-yl)methyl)urea (Chemical Formula 49)

(221) The compound of Chemical formula 49 was prepared according to the preparation method of Example 27.

(222) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.05 (s, 1H), 8.70 (s, 1H), 8.17 (d, J=8.6 Hz, 1H), 7.65 (s, 1H), 7.55 (s, 1H), 7.49-7.37 (m, 2H), 7.26-7.18 (m, 1H), 7.04 (d, J=8.7 Hz, 1H), 6.81 (s, 1H), 6.67 (d, J=7.0 Hz, 1H), 5.61-5.40 (m, 1H), 4.44 (s, 2H), 3.79 (s, 3H), 2.93-2.78 (m, 1H), 2.76-2.63 (m, 2H), 2.10-1.93 (m, 1H).

Example 30

Synthesis of 3-(5-hydroxy-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 57)

(223) ##STR00046##

[30-1] Step 1 Reaction

(224) The mixed solution of 2-hydroxy-6-nitrobenzoic acid (100 mg, 0.55 mmol), 2-aminoglutaimide hydrochloride (180 mg, 1.1 mmol), EDCl-HCl (116 mg, 0.605 mmol), HOBt (93 mg, 0.605 mmol), and DIEA (0.4 ml, 2.2 mmol) was stirred at a room temperature for 30 minutes. The reactants were separated through water and ethyl acetate, and then the organic layer was washed with salt water, and the remained water was dried by anhydrous magnesium sulfate and it was concentrated under reduced pressure, and then was separated and purified by column chromatography to obtain compound <30-1> (41 mg, 25%).

(225) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 10.85 (s, 1H), 8.83 (d, J=7.3 Hz, 1H), 7.50-7.41 (m, 2H), 7.23 (dd, J=7.6, 1.6 Hz, 1H), 4.78-4.70 (m, 1H), 2.79-2.68 (m, 1H), 2.59-2.54 (m, 1H), 2.14-2.09 (m, 1H), 1.96-1.85 (m, 1H).

[30-2] Step 2 Reaction

(226) After dissolving compound <30-1> (34 mg) in the mixed solution of methanol/dimethylformamide (5/1) (6 ml), it was reacted under 10% Pd/C (15 mg) and hydrogen at a room temperature for 2 hours. The reactants were filtered by Celite, and then the filtrates were concentrated to obtain desired JYR-17-187 (22 mg, 70%, white solid).

(227) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.88 (s, 1H), 10.55 (s, 2H), 8.98 (s, 1H), 6.89 (t, J=8.0 Hz, 1H), 6.62 (s, 2H), 6.17 (d, J=8.1 Hz, 1H), 6.04 (d, J=7.8 Hz, 1H), 4.72-4.67 (m, 2H), 2.79-2.72 (m, 1H), 2.55-2.53 (m, 1H), 2.20-2.15 (m, 1H), 2.03-1.95 (m, 1H).

[30-3] Step 3 Reaction

(228) After dissolving compound <30-2> (20 mg, 0.076 mmol) in acetic acid (2 ml), sodium nitrite (9 mg, 0.13 mmol) was added, and then it was stirred at a room temperature for 1 hour. The reactants were separated through water and ethyl acetate, and then the organic layer was washed with salt water, and the remained water was dried by anhydrous magnesium sulfate and it was concentrated under reduced pressure, and then was separated and purified by column chromatography to obtain the compound of Chemical formula 57 (2.7 mg, 13%, ivory solid).

(229) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 11.24 (s, 1H), 10.85 (s, 1H), 8.00 (t, J=8.2 Hz, 1H), 7.72 (d, J=8.0 Hz, 1H), 7.35 (d, J=8.3 Hz, 1H), 5.99-5.95 (m, 1H), 3.01-2.93 (m, 1H), 2.74-2.65 (m, 2H), 2.33-2.28 (m, 1H).

Example 31

Synthesis of (3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)-2,6-dioxopiperidin-1-yl)methyl pivalate (Chemical Formula 58)

(230) ##STR00047##

(231) After dissolving 3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (20 mg, 0.066 mmol) and cesium carbonate (30 mg, 0.099 mmol) in dimethylforamide, chloromethyl pivalate (0.014 ml, 0.099 mmol) was added, and it was stirred at a room temperature for 20 minutes. The reaction for reactants was completed by adding water, and it was extracted with ethyl acetate. The organic layer was washed with salt water and the remained water was dried by anhydrous magnesium sulfate and it was concentrated under reduced pressure, and then was separated and purified by column chromatography to obtain the targeted compound of Chemical formula 58.

(232) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.65 (t, J=8.0 Hz, 1H), 7.37 (dd, J=7.8, 0.9 Hz, 1H), 6.88 (dd, J=8.3, 0.9 Hz, 1H), 6.14 (s, 2H), 5.88 (s, 2H), 5.82-5.74 (m, 1H), 3.17-2.87 (m, 3H), 2.45-2.32 (m, 1H), 1.21 (s, 9H).

Example 32

Synthesis of (3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)-2,6-dioxopiperidin-1-yl)methyl benzoate (Chemical Formula 59)

(233) ##STR00048##

(234) By the same method as the method of Example 31, except for using chloromethyl benzoate instead of chloromethyl pivalate, the compound of Chemical formula 59 was synthesized (17 mg, 63%).

(235) .sup.1H NMR (300 MHz, Chloroform-d) δ 8.06-7.99 (m, 2H), 7.62 (t, J=8.0 Hz, 1H), 7.59-7.52 (m, 1H), 7.46-7.39 (m, 2H), 7.34 (dd, J=7.8, 0.9 Hz, 1H), 6.85 (dd, J=8.3, 0.9 Hz, 1H), 6.19-6.03 (m, 4H), 5.83-5.74 (m, 1H), 3.19-2.85 (m, 3H), 2.45-2.33 (m, 1H).

Example 33

Synthesis of (3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)-2,6-dioxopiperidin-1-yl)methyl piperidine-4-carboxylate hydrochloride (Chemical Formula 60)

(236) ##STR00049##

[33-1] Step 1 Reaction

(237) By the same method as the method of Example 31, except for using 1-(tert-butyl)-4-(chloromethyl)piperidine-1,4-dicarboxylate instead of chloromethyl pivalate, the compound was synthesized (16 mg, 51%, yellow solid).

(238) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.65 (t, J=8.0 Hz, 1H), 7.36 (dd, J=7.8, 0.9 Hz, 1H), 6.88 (dd, J=8.3, 0.9 Hz, 1H), 6.17 (s, 2H), 5.90 (s, 2H), 5.79-5.70 (m, 1H), 4.01 (d, J=13.4 Hz, 2H), 3.17-3.06 (m, 1H), 2.93 (d, J=3.6 Hz, 1H), 2.85 (t, J=13.0 Hz, 2H), 2.54-2.35 (m, 2H), 1.88 (d, J=13.0 Hz, 2H), 1.74-1.64 (m, 2H), 1.47 (s, 9H).

[33-2] Step 2 Reaction

(239) After dissolving the compound obtained in Step 1 in dichloromethane, 4N HCl 1,4-dioxene solution was added, and it was stirred at a room temperature for 1 hour. The reactants were concentrated to obtain the targeted compound of Chemical formula 60 (80%).

(240) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.63 (t, J=8.0 Hz, 1H), 7.37-7.33 (m, 1H), 6.89-6.81 (m, 1H), 6.14 (s, 2H), 5.87 (s, 2H), 5.78-5.72 (m, 1H), 3.82-3.58 (m, 1H), 3.19-3.01 (m, 3H), 2.99-2.82 (m, 2H), 2.70-2.57 (m, 2H), 2.55-2.29 (m, 2H), 1.89 (d, J=13.3 Hz, 2H), 1.75-1.55 (m, 2H).

Example 34

Synthesis of (3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)-2,6-dioxopiperidin-1-yl)methyl D-prolinate hydrochloride (Chemical Formula 61) and 2-((3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)-2,6-dioxopiperidin-1-yl)methyl) 1-(tert-butyl) (2R)-pyrrolidine-1,2-dicarboxylate (Chemical Formula 64)

(241) ##STR00050##

[34-1] Step 1 reaction

(242) By the same method as the method of Example 31, except for using 1-(tert-butyl)2-(chloromethyl)(R)-pyrrolidine-1,2-dicarboxylate instead of chloromethyl pivalate, the compound of Chemical formula 64 was synthesized.

(243) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.66 (t, J=8.0 Hz, 1H), 7.37 (d, J=7.8 Hz, 1H), 6.89 (d, J=8.2 Hz, 1H), 6.15 (s, 2H), 5.93 (s, 1H), 5.80-5.69 (m, 1H), 3.15-2.85 (m, 4H), 2.38 (d, J=6.6 Hz, 1H), 2.23-1.99 (m, 1H), 1.91 (dd, J=13.5, 7.1 Hz, 1H), 1.84-1.78 (m, 3H).

[34-2] Step 2 Reaction

(244) After dissolving the compound obtained in the step 1 in dichloromethane, 4N HCl 1,4-dioxene solution was added, and it was stirred at a room temperature for 1 hour. The reactants were concentrated to obtain the targeted compound of Chemical formula 61.

(245) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.66 (t, J=8.0 Hz, 1H), 7.37 (d, J=7.8 Hz, 1H), 6.89 (d, J=8.2 Hz, 1H), 6.15 (s, 2H), 5.93 (s, 1H), 5.80-5.69 (m, 1H), 3.15-2.85 (m, 4H), 2.38 (d, J=6.6 Hz, 1H), 2.23-1.99 (m, 1H), 1.91 (dd, J=13.5, 7.1 Hz, 1H), 1.84-1.78 (m, 3H).

Example 35

Synthesis of (3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)-2,6-dioxopiperidin-1-yl)methyl L-isoleucinate hydrochloride (Chemical Formula 62)

(246) ##STR00051##

[35-1] Step 1 Reaction

(247) By the same method as the method of Example 31, except for using (tert-butoxy carbonyl)-D-isoleucinate instead of chloromethyl pivalate, the compound was synthesized.

[35-2] Step 2 Reaction

(248) After dissolving the compound obtained in the step 1 in dichloromethane, 4N HCl 1,4-dioxene solution was added, and it was stirred at a room temperature for 1 hour. The reactants were concentrated to obtain the targeted compound of Chemical formula 62.

(249) .sup.1H NMR (300 MHz, Chloroform-d) δ 7.69-7.61 (m, 1H), 7.36 (d, J=7.8 Hz, 1H), 6.88 (d, J=8.3 Hz, 1H), 6.16 (s, 2H), 5.92 (s, 2H), 5.83-5.71 (m, 1H), 3.37 (d, J=5.0 Hz, 1H), 3.18-2.82 (m, 3H), 2.48-2.30 (m, 1H), 1.54-1.38 (m, 1H), 1.27-1.08 (m, 2H), 0.95 (d, J=6.8 Hz, 2H), 0.94-0.85 (m, 4H).

Example 36

Synthesis of (3-(5-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)-2,6-dioxopiperidin-1-yl)methyl butyrate (Chemical Formula 63)

(250) ##STR00052##

(251) By the same method as the method of Example 31, except for using chloromethyl butyrate instead of chloromethyl pivalate, the compound of Chemical formula 63 was synthesized (44%, yellow solid).

(252) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.62 (t, J=8.1 Hz, 1H), 7.37-7.31 (m, 1H), 6.89-6.82 (m, 1H), 6.14 (s, 2H), 5.86 (d, J=2.2 Hz, 2H), 5.81-5.71 (m, 1H), 3.16-3.03 (m, 1H), 2.98-2.86 (m, 2H), 2.44-2.33 (m, 1H), 2.29 (t, J=7.4 Hz, 2H), 1.71-1.63 (m, 2H), 0.93 (t, J=7.4 Hz, 3H).

Example 37

Synthesis of 3-(6-amino-4-oxobenzo[d][1,2,3]triazin-3(4H)-yl)piperidine-2,6-dione (Chemical Formula 65)

(253) The compound of Chemical formula 65 was prepared according to the preparation method of Example 7.

(254) .sup.1H NMR (500 MHz, Methanol-d.sub.4) δ 7.88 (d, J=10.0 Hz, 1H), 7.30-7.28 (m, 2H), 5.87 (m, 1H), 3.02-2.79 (m, 3H), 2.39-2.32 (m, 1H).

Test Example 1

(255) Evaluation of Substrate Proteolytic Activity Against CRBN

(256) In order to confirm whether the compound of the present disclosure could inhibit the function of CRBN by specifically binding to CRBN (celebron), the effect of the present compound on the proteolytic activity of Ikaros/Aiolos protein (Nat Struct Mol Biol. 2014 September; 21(9):803-9) or GSPT1 protein (Nature 535, 252257 (14 Jul. 2016)), degraded by CRBN protein when binding to thalidomide and its derivative, was investigated.

(257) Pomalidomide or CC-885 compound which inhibited the function by binding to CRBN was used as a positive control group.

(258) To evaluate the degradative activity of Ikaros/Aiolos protein or GSPT1 protein of the compound according to the present disclosure, an experiment was conducted as follows.

(259) 5×10.sup.5 OCI-LY3 cells were seeded on a 12-well plate, and then each compound was treated to each well in a defined concentration. In 6 hours or 24 hours, cell lysates were collected using TBSN buffer. The proteolytic activity of Ikaros/Aiolos protein was evaluated using an antibody against Aiolos protein, and the proteolytic activity of GSPT1 was evaluated by western blot using an antibody against GSPT1 protein, and for this, the same amount of protein was loaded on each well of 4-15% gradient gel and then after electrophoresis, the protein was transferred to PVDF membrane, and the primary antibody against each protein was bound. After that, the secondary antibody in which HRP was attached was bound, and it was developed using HRP substrate.

(260) As a result, as shown in FIG. 1, it could be seen that when treating the compound of the present disclosure, Ikaros/Aiolos protein was degraded in a concentration dependent manner, compared to the group treated with the negative control group, DMSO, and it tended to be similar to the positive control group, pomalidomide.

(261) In addition, as shown in FIG. 2, as a result of treating for 6 hours or 24 hours, respectively, in the OCI-LY3 cell line, it could be confirmed that the compound of the present disclosure facilitated the degradation of GSPT1 protein and Aiolos protein.

Test Example 2

(262) Cytotoxicity Experiment

(263) In order to confirm the effect of the compound of the present disclosure on cancer cells and normal cells, the cytotoxicity experiment was conducted as follows.

(264) After seeding NCI-H929 cells on a 96-well plate so as to be 10,000 cells per each well, each compound was treated in a predetermined concentration. In 72 hours, WST-1 reagent was added, and in 1 hour, the absorbance was measured at 450 nm using spectramax spectrophotometer to measure the degree of cell death. Using the measured value, IC.sub.50 was calculated with graphpad prism program, and this was shown in Table 1.

(265) TABLE-US-00001 TABLE 1 Cytotoxicity Compound number activity Compound of Chemical A formula 7 Compound of Chemical B formula 8 Compound of Chemical C formula 9 Compound of Chemical B formula 11 Compound of Chemical C formula 12 Compound of Chemical C formula 19 Compound of Chemical C formula 20 Compound of Chemical C formula 21 Compound of Chemical C formula 23 Compound of Chemical C formula 24 Compound of Chemical B formula 25 Compound of Chemical C formula 26 Compound of Chemical C formula 27 Compound of Chemical C formula 29 Compound of Chemical C formula 30 Compound of Chemical C formula 31 Compound of Chemical A formula 37 Compound of Chemical A formula 38 Compound of Chemical C formula 56 Compound of Chemical A formula 57 Compound of Chemical A formula 58 Compound of Chemical A formula 59 Compound of Chemical A formula 60 Compound of Chemical A formula 61 Compound of Chemical A formula 62 Compound of Chemical A formula 63 Compound of Chemical C formula 65 * IC.sub.50 value - A: <1 μM, B: 1-10 μM, C: 10-50 μM, D: >50 μM

(266) As a result, as shown in Table 1, it could be seen that the compounds of the present disclosure had sufficient cytotoxicity in the tumor cell, NCI-H929.

(267) In particular, it could be confirmed that Compounds 7˜9, 11, 12, 19˜21, 23, 25˜27, 29˜31, 37, 38, 56˜63 and 65 as triazine derivative compounds wherein X is N in the Chemical formula 1 of the present disclosure had the excellent cytotoxicity activity against cancer cells, compared to Compounds 24 and 31 as pyridine derivative compounds wherein X is C.