KR101034051B1 - Novel isoindolo[2,1-b]isoquinolinone derivatives and anti-cancer agents including them - Google Patents

Abstract

The present invention relates to an isoindolo [2,1-b] isoquinolinone derivative of Formula 1 and an anticancer composition containing the same, wherein the isoindolo [2,1-b] isoquinolinone derivative of the present invention is a cancer cell It can be usefully used as an inhibitor of topoisomerase I of.

[Formula 1]

[Wherein, R ₁ to R ₄ and R ₁₁ to R ₁₄ are as defined in the detailed description of the invention.]

Isoindolo [2,1-b] isoquinolinone derivatives, anticancer agents, topoisomerase I

Description

Novel isoindolo [2,1-b] isoquinolinone derivatives and anti-cancer agents including them}

The present invention relates to an isoindolo [2,1-b] isoquinolinone derivative of Formula 1 and an anticancer composition containing the same, wherein the isoindolo [2,1-b] isoquinolinone derivative of the present invention is a cancer cell It can be usefully used as an inhibitor of topoisomerase I of.

[Formula 1]

[In Formula 1, R ₁ to R ₄ are independently of each other hydrogen, halogen or (C1-C7) alkyl, and R ₁₁ to R ₁₄ are independently of each other hydrogen, halogen or (C1-C7) alkoxy.]

Camptothecin (S) -4-Ethyl-4-hydroxy-1H-pyrano [3 ', 4': 6,7] indoli zino [1,2-b] quinoline-3,14 (4H, 12H) -dione) is a pyrrole [3,4-b] quinoline alkaloid having the following structure and is known as a potent anticancer agent having antitumor activity by binding to and inhibiting topoisomerase I. The topoisomerase I is a nuclear enzyme that reduces torsional stress during DNA replication and plays an important role in DNA replication.

The camptothecin has a pentacyclic structure having a S-type chiral center at the 20-position of ring E and a lactone structure near the chiral center. Although camptothecin has a definite therapeutic effect on gastric cancer and rectal cancer, clinical studies on it have been limited due to its low solubility and toxicity in water.

By altering the molecular structure of the camptothecin, intensive studies have been made to obtain camptothecin derivatives with higher activity and lower toxicity. Accordingly, numerous camptothecin compounds with good efficacy were synthesized. In addition, it has been found that the anticancer activity of camptothecin can be increased by introducing an appropriate group at the 9-position of 10-hydroxy camptothecin. For example, Topotecan, which is currently commonly used, is the compound.

Therefore, the present inventors found that isoindolo [2,1-b] isoquinolinone derivatives having a skeleton similar to camptothecin have topoisomerase I inhibitory activity and are useful as a cancer-causing agent because they have cytotoxicity against human cancer cells. The present invention was completed by finding out.

It is an object of the present invention to provide novel isoindolo [2,1-b] isoquinolinone derivatives having excellent anticancer effects and pharmaceutically acceptable salts thereof.

Another object of the present invention is to provide an anticancer agent composition containing the isoindolo [2,1-b] isoquinolinone derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention relates to an isoindolo [2,1-b] isoquinolinone derivative of Formula 1 and an anticancer composition containing the same, wherein the isoindolo [2,1-b] isoquinolinone derivative of the present invention is a cancer cell It can be usefully used as an inhibitor of topoisomerase I of.

[Formula 1]

[In Formula 1, R ₁ to R ₄ are independently of each other hydrogen, halogen or (C1-C7) alkyl, and R ₁₁ to R ₁₄ are independently of each other hydrogen, halogen or (C1-C7) alkoxy.]

R ₁ and R ₂ are independently of each other hydrogen, chloro, fluorine or methyl, and R ₁₁ and R ₁₂ are independently of each other hydrogen, chloro, fluorine or methoxy.

Preferred compounds of the isoindolo [2,1-b] isoquinolinone derivative of the general formula (I) of the present invention are as follows:

The isoindolo [2,1-b] isoquinolinone derivatives of the general formula (1) of the present invention are prepared by intramolecular ring addition reaction of the hydroxymethyl compound of the general formula (2).

[Formula 1]

[Formula 2]

[In Formulas 1 and 2, R ₁ to R ₄ are independently of each other hydrogen, halogen or (C1-C7) alkyl, and R ₁₁ to R ₁₄ are independently of each other hydrogen, halogen or (C1-C7) alkoxy. ]

In addition, the hydroxymethyl compound of Formula 2 is prepared by reacting the compound of Formula 3 with DDQ (2,3-dichloro-5,6-dicyano-l, 4-benzoquinone).

(3)

[In the above formula, R ₁ to R ₄ are independently of each other hydrogen, halogen or (C1-C7) alkyl, R ₁₁ to R ₁₄ are independently of each other hydrogen, halogen or (C1-C7) alkoxy, R ₁₅ is (C1-C7) alkoxy (C1-C7) alkyl or (C1-C7) alkoxy (C6-C12) aryl.]

In addition, the compound of Formula 3 is prepared by reacting an o-toluimide compound of Formula 4 with a benzonitrile compound of Formula 5.

[Formula 4]

[Chemical Formula 5]

[In the above formula, R ₁ to R ₄ are independently of each other hydrogen, halogen or (C1-C7) alkyl, R ₅ and R ₆ are independently of each other hydrogen or (C1-C7) alkyl, R ₁₁ to R ₁₄ Are independently of each other hydrogen, halogen or (C1-C7) alkoxy, and R ₁₅ is (C1-C7) alkoxy (C1-C7) alkyl or (C1-C7) alkoxy (C6-C12) aryl.]

The isoindolo [2,1-b] isoquinolinone derivatives of formula 1 of the present invention may be used in the form of pharmaceutically acceptable salts derived from inorganic or organic acids according to methods well known in the art, Hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, acetic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, mandelic acid, tartaric acid, citric acid, ascorbic acid, palmitic acid, maleic acid And salts such as hydroxymaleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, salicylic acid, methanesulfonic acid, benzenesulfonic acid and toluenesulfonic acid.

In the present invention, the compound of formula 1 inhibits the activity of Topo-I and exhibits cytotoxicity against cancer cell lines such as lung cancer cell line A549, colon cancer cell line HCT15, and ovarian cancer cell line OV-3, and the like. Pharmaceutical compositions containing a compound of or a pharmaceutically acceptable salt thereof can be used for the prevention or treatment of various cancers, including lung cancer, colorectal cancer and ovarian cancer.

The pharmaceutical compositions of the invention can be formulated in a variety of oral or parenteral dosage forms. Formulations for oral administration include, for example, tablets, pills, hard and soft capsules, solutions, suspensions, emulsifiers, syrups, and granules. These formulations may contain diluents (e.g., lactose, dextrose, water, etc.) in addition to the active ingredients. Cross, mannitol, sorbitol, cellulose and / or glycine) and glidants such as silica, talc, stearic acid and its magnesium or potassium salts and / or polyethylene glycols. Tablets may also contain binders such as magnesium aluminum silicate and / or polyvinylpyrrolidine, optionally disintegrating or boiling mixtures such as starch, agar, alginic acid or its sodium salt and / or absorbents, colorants, flavors And sweeteners. The formulations may be prepared by conventional mixing, granulating or coating methods. Also representative of formulations for parenteral administration are injectable formulations, preferably isotonic solutions or suspensions.

The composition may contain sterile and / or auxiliaries such as preservatives, stabilizers, hydrating or emulsifying accelerators, salts and / or buffers for the control of osmotic pressure, and other therapeutically useful substances, which are conventional methods of mixing, granulating Or according to a coating method. As active ingredients, the compounds of formula 1 may be administered via oral or parenteral routes once daily or in divided doses, in an amount of 0.01 to 10 mg / kg body weight per day for mammals, including humans.

Isoindolo [2,1-b] isoquinolinone derivatives according to the present invention are novel compounds and are efficiently prepared through intramolecular ring addition reactions. In addition, the isoindolo [2,1-b] isoquinolinone derivatives according to the present invention have an excellent inhibitory effect on topoisomerase and also have excellent cytotoxicity against human cancer cells, and thus can be usefully used as a cancer-containing agent.

Hereinafter, the present invention will be described in more detail with reference to Preparation Examples, Examples and Experimental Examples. However, the following Preparation Examples and Examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.

[ Manufacturing example  1] 2,4, N- Trimethylbenzamide (2,4, N- Trimethylbenzamide ) Preparation of (5b)

2,4-dimethylbenzoic acid (2 g, 13.3 mmol) and pyridine (1.7 mL) were suspended in CH ₂ Cl ₂ (50 mL) and then oxalyl chloride (5.7 mL, 66.6 mmol) was slowly added with stirring at 0 ° C. Was added. After stirring for another 2 hours, excess oxalyl chloride was removed by co-distillation with benzene. The residue was dissolved in CH ₂ Cl ₂ (50 mL) and 40% methylamine (17 mL, 199 mmol) was added at 0 ° C. After stirring overnight, the reaction mixture was diluted with water, the organic layer was separated and the water layer was extracted with CH ₂ Cl ₂ . The organic layer was washed with water and brine, dried and concentrated. The concentrated residue was purified by column chromatography (n-hexane: EA = 2: 1) to give 2,4, N-trimethylbenzamide ( 5b ) (white solid, 1 g, 46%).

Mp: 100 102 ° C. IR (cm- ¹ ): 3289 (NH), 1636 (C = O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 7.22 (d, J = 7.7 Hz, 1H), 7.01 (s, 1H), 6.97 (d, J = 7.7 Hz, 1H), 5.80 (s, 1H), 2.95 (d, J = 4.9 Hz, 3H), 2.39 (s, 3H), 2.31 (s, 3H).

[ Manufacturing example  2] 2,5, N- Trimethylbenzamide (2,5, N- Trimethylbenzamide ) Preparation of (5c)

2,5, N-trimethylbenzamide (2,5-dimethylbenzoic acid (5 g, 33.3 mmol), pyridine (4.5 mL) and oxalyl chloride (21.1 g, 166 mmol) in the same manner as in Preparation Example 1 5c ) (white solid, 2.7 g, 50%).

Mp: 125.1-126.3 ° C. IR (cm- ¹ ): 3289 (NH), 1636 (C = O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 7.15 (s, 1H), 7.09 (s, 2H), 5.80 (s, 1H), 2.97 (d, 3H), 2.38 (s, 3H), 2.30 (s , 3H).

[ Manufacturing example  3] N, N- Diethyl -2- Methylbenzamide (N, N- Diethyl -2- methylbenzamide ) Manufacturing of (5d)

To thionyl chloride (68 mL) in an ice-bath o -toluic acid (13.6 g, 100 mmol) was added at a ratio (portionwise). After removing the ice-bath, the reaction mixture was warmed to 50 ° C. for 45 minutes to completely dissolve o -toluic acid. The reaction mixture was slowly cooled to room temperature with stirring overnight. Excess thionyl chloride and reaction byproducts HCl and SO ₂ were removed by vacuum distillation. The residue was dissolved in CH ₂ Cl ₂ (100 mL) and diethylamine (78 mL, 760 mmol) was added at 0 ° C. After stirring overnight, the reaction mixture was diluted with water, the organic layer was separated and the water layer was extracted with CH ₂ Cl ₂ . The organic layer was washed with water and brine, dried and concentrated. The concentrated residue was purified by column chromatography (n-hexane: EA = 1: 1) to give N, N-diethyl-2-methylbenzamide ( 5d ) (yellow oil, 18.7 g, 98%).

IR (cm- ¹ ): 1631 (C = O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 7.23-7.13 (m, 4H); 3.80 (s, 1 H); 3.40 (s, 1 H); 3.09 (q, J = 6.9 Hz, 2H); 2.27 (s, 3 H); 1.24 (t, J = 6.9 Hz, 3H); 0.99 (t, J = 7.2 Hz, 3H).

[ Manufacturing example  4] N, N- Diethyl -2,4-dimethylbenzamide (N, N- Diethyl Preparation of -2,4-dimethylbenzamide) (5e)

N, N-diethyl-2 in the same manner as in Preparation Example 3 using 2,5-dimethylbenzoic acid (10 g, 66.7 mmol), thionyl chloride (45 mL) and diethylamine (69 mL, 667 mmol) , 4-dimethylbenzamide ( 5e ) was obtained (light yellow oil, 12.5 g, 92%).

IR (cm- ¹ ): 1632 (C = O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 7.06-7.01 (m, 3H), 3.60 (s, 2H); 3.12 (q, J = 7.1 Hz, 2H); 2.31 (s, 3 H); 2.24 (s, 3 H); 1.25 (t, J = 7.1 Hz, 3H); 1.02 (t, J = 7.1 Hz, 3H).

[ Manufacturing example  5] N, N- Diethyl -2,5-dimethylbenzamide (N, N- Diethyl -2,5-dimethylbenzamide) (5f) Preparation

N, N-diethyl-2 in the same manner as in Preparation Example 3 using 2,5-dimethylbenzoic acid (7.5 g, 50 mmol), thionyl chloride (50 mL) and diethylamine (69 mL, 667 mmol) , 5-dimethylbenzamide ( 5f ) was obtained (light yellow oil, 9.6 g, 94%).

IR (cm- ¹ ): 1632 (C = O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 7.05 (s, 2H); 6.96 (s, 1 H); 3.75 (s, 1 H); 3.40 (s, 1 H); 3.15-3.09 (m, 2 H); 2.29-2.20 (d, 6 H); 1.25 (t, J = 7.1 Hz, 3H); 1.02 (t, J = 7.1 Hz, 3H).

[ Manufacturing example  6] 4- Methoxy -2-(( Methoxy methoxy ) methyl ) Benzonitrile Preparation of (4-Methoxy-2-((methoxymethoxy) methyl) benzonitrile) (6b)

Diisopropylethylamine (DIPEA) (6.53 g, 50 mmol) in a mixture of 4-methoxy-2-hydroxymethylbenzonitrile (4.08 g, 25 mmol) and CH ₂ Cl ₂ (20 mL) at 0 ° C. And chloromethylmethyl ether (4.02 g, 50 mmol) were added. After completion of the reaction, CH ₂ Cl ₂ was removed in vacuo and the residue was purified by column chromatography (n-hexane: EA = 3: 1) to give 4-methoxy-2-((methoxymethoxy) methyl) Obtained benzonitrile ( 6b ) (yellow oil, 4.70 g, 91%).

IR (cm- ¹ ): (CN). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 7.59 (d, J = 8.4 Hz, 1H); 7.10 (d, J = 2.6 Hz, 1H); 6.90 (dd, J ₁ = 2.6 Hz, J ₂ = 8.6 Hz, 1H); 4.77 (s, 2 H); 4.74 (s, 2 H); 3.87 (s, 3 H); 3.44 (s, 3 H).

[ Manufacturing example  7] 4,5- Dimethoxy -2-(( Methoxy methoxy ) methyl ) Benzonitrile (4,5- Dimethoxy -2-(( methoxymethoxy ) methyl ) benzonitrile ) Preparation of (6c)

Preparation Example 6 using 4,5-dimethoxy-2-hydroxymethylbenzonitrile (5.5 g, 28.5 mmol), DIPEA (7.35 g, 57 mmol) and chloromethylmethyl ether (4.59 g, 57 mmol) In the same manner, 4,5-dimethoxy-2-((methoxymethoxy) methyl) benzonitrile ( 6c ) was obtained (white solid, 6.7 g, 99%).

Mp: 54.5-56.4 ° C. IR (cm ⁻¹ ) :( CN). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 7.07 -7.03 (d, 2H); 4.76 (s, 2 H); 4.71 (s, 2 H); 3.95 (s, 3 H); 3.91 (s, 3 H); 3.44 (s, 3 H).

[ Manufacturing example  8] 3- [2- (4- Methoxybenzyloxymethyl ) Phenyl ] -2H-isoquinolin-1-one (3- [2- (4- Methoxybenzyloxymethyl ) phenyl ] -2H- isoquinolin -One- one ) Production of (7a)

N - methyl-o- toluamide (5.03 g, 33.7 mmol) and dry THF (50㎖) n -BuLi ( 2.5M in n -hexane) in a nitrogen atmosphere to a solution, -20 ℃ to mix (22.8 ml, 57 mmol) was added and maintained at a temperature not exceeding 0 ° C. After the addition was completed, the mixture was stirred at the same temperature for 1 hour. A solution of 2-{[( p -methoxybenzyl) oxy] methyl} benzonitrile (26.9 mmol, 6.82 g) and dry THF (30 mL) was slowly added to the solution at -50 ° C and at the same temperature. Stir for 20 minutes. The reaction mixture was quenched with water, stirred vigorously for 10 minutes, and extracted with EA. The organic layer was washed with water and brine and dried over anhydrous Na ₂ SO ₄ . After removing the solvent, the residue was purified by column chromatography (n-hexane: EA = 3: 1) to give 3- [2- (4-methoxybenzyloxymethyl) phenyl] -2H-isoquinolin-1-one ( 7a ) (yellow oil, 2.74 g, 44%).

IR (cm- ¹ ): 3400 (NH), 1657 (C = O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 10.25 (s, 1H); 8.44 (m, 1 H); 7.65 (m, 1 H); 7.57-7.25 (m, 8 H); 6.90 (m, 2 H); 6.58 (s, 1 H); 4.64 (s, 2 H); 4.46 (s, 2 H); 3.77 (s, 3 H).

[ Manufacturing example  9] 3- [2- (4- Methoxybenzyloxymethyl ) Phenyl ] -6- methyl -2H-isoquinolin-1-one (3- [2- (4- Methoxybenzyloxymethyl ) phenyl ] -6- methyl -2H- isoquinolin -One- one ) Production of (7b)

Using Compound 5b (1 g, 6.1 mmol) and 2-{[( p -methoxybenzyl) oxy] methyl} benzonitrile (7.7 mmol, 1.95 g) of Preparation Example 3, -[2- (4-methoxybenzyloxymethyl) phenyl] -6-methyl-2H-isoquinolin-1-one ( 7b ) was obtained (yellow oil, 892 mg, 38%).

IR (cm- ¹ ): 3400 (NH), 1657 (C = O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 10.12 (s, 1H), 8.33 (d, 1H), 7.55- 6.86 (m, 10H), 6.53 (s, 1H), 4.64 (s, 2H), 4.46 (s, 2H), 3.79 (s, 3H), 2.50 (s, 3H).

[ Manufacturing example  10] 3- [2- (4- Methoxybenzyloxymethyl ) Phenyl ] -7- methyl -2H-isoquinolin-1-one (3- [2- (4- Methoxybenzyloxymethyl ) phenyl ] -7- methyl -2H- isoquinolin -One- one ) Preparation of (7c)

Using Compound 5c (1.5 g, 9.2 mmol) and 2-{[( p -methoxybenzyl) oxy] methyl} benzonitrile (11.8 mmol, 3 g) of Preparation Example 3 in the same manner as in Preparation Example 3 -[2- (4-methoxybenzyloxymethyl) phenyl] -7-methyl-2H-isoquinolin-1-one ( 7c ) was obtained (yellow oil, 1.85 g, 52%).

IR (cm- ¹ ): 3400 (NH), 1657 (C = O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 10.2 (s, 1H); 8.25 (s, 1 H); 7.54-7.41 (m, 8 H); 6.90 (m, 2 H); 6.56 (s, 1 H); 4.64 (s, 2 H); 4.45 (s, 2 H); 3.78 (s, 3 H); 2.50 (s, 3 H).

[ Manufacturing example  11] 3- (4- Methoxy -2-(( Methoxy methoxy ) methyl ) Phenyl )-2 H Isoquinolin-1-one (3- (4- Methoxy -2-(( methoxymethoxy ) methyl ) phenyl )-2 H - isoquinolin -One- one ) Manufacturing of (7d)

N-BuLi (6 mL of 2.5 M in hexane) in a solution obtained by mixing compound 5d (1.68 g, 8.8 mmol) of Preparation Example 3 and compound 6b (1.52 g, 7.3 mmol) of Preparation Example 6 in dry THF (20 mL). , 15 mmol) and THF (20 mL) were added dropwise at -70 ° C. The reaction mixture was stirred for 6 hours at the same temperature. The reaction was quenched with water, extracted with EA and dried over anhydrous Na ₂ SO ₄ . After removal of the solvent, the residue was purified by column chromatography (n- hexane: EA = 1: 1) 3- (4- methoxy-2 - ((methoxymethoxy) methyl) phenyl) -2, to obtain H-iso Quinolin-1-one ( 7d ) was obtained (yellow oil, 985 mg, 41%).

IR (cm ⁻¹ ) :( NH), 1655 (C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 9.79 (s, 1H); 8.40 (d, 1 H); 7.67 (m, 1 H); 7.56 (m, 1 H); 7.48 (m, 2 H); 7.03 (m, 1 H); 6.97 (m, 1 H); 6.52 (s, 1 H); 4.80 (s, 2 H); 4.56 (s, 2 H); 3.87 (s, 3 H); 3.43 (s, 3 H).

[ Manufacturing example  12] 3- (4,5- Dimethoxy -2-(( Methoxy methoxy ) methyl ) Phenyl )-2 H Isoquinolin-1-one (3- (4,5-Dimethoxy-2-((methoxymethoxy) methyl) phenyl) -2 H -isoquinolin-1-one) (7e)

Using Compound 5d (1.85 g, 9.7 mmol) of Preparation Example 3 and Compound 6c (1.8 g, 7.6 mmol) of Preparation Example 7 in the same manner as in Preparation Example 3, 3- (4,5-dimethoxy-2- ( (methoxymethoxy) methyl) phenyl) -2 H - isoquinolin-1-one was obtained (7e) (yellow solid, 2.0 g, 58%).

Mp: 122.5-124.5 ° C. IR (cm ⁻¹ ) :( NH), 1655 (C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 10.30 (s, 1H); 8.38 (d, J = 8 Hz, 1 H); 7.66 (m, 1 H), 7.57 (d, J = 7.5 Hz, 1 H), 7.48 (m, 1 H), 7.06 (s, 1 H); 7.00 (s, 1 H); 6.59 (s, 1 H); 4.79 (s, 2 H); 4.56 (s, 2 H); 3.97 (d, 6 H); 3.43 (s, 3 H).

[ Manufacturing example  13] 3- (4- Methoxy -2-(( Methoxy methoxy ) methyl ) Phenyl ) -6- methyl -2H-isoquinolin-1-one (3- (4- Methoxy -2-(( methoxymethoxy ) methyl ) phenyl ) -6- methyl -2H- isoquinolin -1-one) (7f) Preparation

Using Compound 5e (1.25 g, 6.1 mmol) of Preparation Example 4 and compound 6b (1.6 g, 7.7 mmol) of Preparation Example 6, 3- (4-methoxy-2-((meth) was prepared in the same manner as in Preparation Example 11. Methoxymethoxy) methyl) phenyl) -6-methyl-2H-isoquinolin-1-one ( 7f ) was obtained (yellow solid, 892 mg, 43%).

Mp: 103-104.1 ° C. IR (cm ⁻¹ ) :( NH), 1655 (C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 10.05 (s, 1H); 8.26 (d, J = 8.2 Hz, 1H); 7.44 (d, J = 8.5 Hz, 1H); 7.30 (m, 2 H); 7.04 (d, J = 2.6 Hz, 1H); 6.94 (dd, J ₁ = 2.7 Hz, J ₂ = 8.4 Hz, 1H); 6.44 (s, 1 H); 4.76 (s, 2 H); 4.55 (s, 2 H); 3.85 (s, 3 H); 3.39 (s, 3 H); 2.47 (s, 3 H).

[ Manufacturing example  14] 3- (4,5- Dimethoxy -2-(( Methoxy methoxy ) methyl ) Phenyl ) -6- methyl -2H-isoquinolin-1-one (3- (4,5- Dimethoxy -2-(( methoxymethoxy ) methyl ) phenyl ) -6- methyl -2H-isoquinolin-1-one) (7 g)

Using Compound 5e (1.1 g, 5.4 mmol) of Preparation Example 4 and Compound 6c (1 g, 4.2 mmol) of Preparation Example 7 in the same manner as in Preparation Example 3- (4,5-dimethoxy-2- ( (Methoxymethoxy) methyl) phenyl) -6-methyl-2H-isoquinolin-1-one ( 7 g ) was obtained (yellow solid, 1.2 g, 77%).

Mp: 126.4-127.8 ° C. IR (cm ⁻¹ ) :( NH), 1655 (C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 9.75 (s, 1H); 8.29 (d, J = 8.1 Hz, 1 H); 7.34 (m, 2 H); 6.99 (d, 2 H); 6.48 (s, 1 H); 4.80 (s, 2 H); 4.53 (s, 2 H); 3.95 (d, 6 H); 3.44 (s, 3 H); 2.49 (s, 3 H).

[ Manufacturing example  15] 3- (4- Methoxy -2-(( Methoxy methoxy ) methyl ) Phenyl ) -7- methyl -2H-isoquinolin-1-one (3- (4- Methoxy -2-(( methoxymethoxy ) methyl ) phenyl ) -7- methyl -2H-  isoquinolin -1-one) (7h) Preparation

Using Compound 5e (820 mg, 4 mmol) of Preparation Example 4 and Compound 6b (1.03 g, 5 mmol) of Preparation Example 6, 3- (4-methoxy-2-((meth) was prepared in the same manner as in Preparation Example 11. Methoxymethoxy) methyl) phenyl) -7-methyl-2H-isoquinolin-1-one ( 7h ) was obtained (yellow solid, 585 mg, 42%).

Mp: 74.0-75.8 ° C. IR (cm ⁻¹ ) :( NH), 1655 (C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 9.80 (s, 1H); 8.19 (s, 1 H); 7.48-7.43 (m, 3 H); 7.03 (d, J = 2.6 Hz, 1H); 6.95 (dd, J ₁ = 2.7 Hz, J ₂ = 8.5 Hz, 1H); 6.49 (s, 1 H); 4.78 (s, 2 H); 4.54 (s, 2 H); 3.86 (s, 3 H); 3.41 (s, 3 H); 2.49 (s, 3 H).

[ Manufacturing example  16] 3- (4,5- Dimethoxy -2-(( Methoxy methoxy ) methyl ) Phenyl ) -7- methyl -2H-isoquinolin-1-one (3- (4,5- Dimethoxy -2-(( methoxymethoxy ) methyl ) phenyl ) -7- methyl -2H-isoquinolin-1-one) (7i)

Using compound 5f (1.1 g, 5.4 mmol) of Preparation Example 5 and compound 6c (1 g, 4.2 mmol) of Preparation Example 7 in the same manner as in Preparation Example 3- (4,5-dimethoxy-2- ( (Methoxymethoxy) methyl) phenyl) -7-methyl-2H-isoquinolin-1-one ( 7i ) was obtained (yellow solid, 730 mg, 47%).

Mp: 135.5-137.1 ° C. IR (cm ⁻¹ ) :( NH), 1655 (C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 9.75 (s, 1H); 8.21 (s, 1 H); 7.50 (m, 2 H); 6.99 (d, 2 H); 6.53 (s, 1 H); 4.80 (s, 2 H); 4.53 (s, 2 H); 3.95 (d, 6 H); 3.44 (s, 3 H); 2.50 (s, 3 H).

[ Manufacturing example  16] 3- (2- Hydroxymethylphenyl Preparation of 2-H-isoquinolin-1-one (3- (2-Hydroxymethylphenyl) -2H-isoquinolin-1-one) (8a)

DDQ (2.56 g, 11.1 mmol) was added to a solution of compound 7a (2.74 g, 7.4 mmol) and water (2 mL) / CH ₂ Cl ₂ (36 mL) of Preparation Example 8 at room temperature. The reaction mixture was stirred overnight, then saturated aqueous NaHCO ₃ solution (20 mL) was added and extracted with methylene chloride. The organic layer was washed with water and brine, dried over anhydrous Na ₂ SO ₄ , and concentrated under reduced pressure. The residue was purified by column chromatography (n-hexane: EA = 2: 1) to give 3- (2-hydroxymethylphenyl) -2H-isoquinolin-1-one ( 8a ) (brown solid, 1.66 g, 89 %).

Mp: 175 177 ° C. IR (cm ⁻¹ ) :( OH), 1629 (C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 11.28 (s, 1H); 8.06 (d, J = 8.1 Hz, 1 H); 7.56-7.21 (m, 7H); 6.42 (s, 1 H); 5.33 (m, 1 H); 4.39 (m, 2 H).

[ Manufacturing example  17] 3- (2- Hydroxymethylphenyl ) -6- methyl Preparation of -2H-isoquinolin-1-one (3- (2-Hydroxymethylphenyl) -6-methyl-2H-isoquinolin-1-one) (8b)

3- (2-hydroxymethylphenyl) -6-methyl-2H-isoquinoline in the same manner as in Preparation 16 using compound 7b (892 mg, 2.3 mmol) and DDQ (794 mg, 3.5 mmol) in Preparation Example 9 Obtained -1-one ( 8b ) (yellow solid, 285 mg, 47%) .

Mp: 174.5 181.6 ° C. IR (cm ⁻¹ ) :( OH), 1629 (C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 11.30 (s, 1H); 8.19 (d, J = 8.1 Hz, 1 H); 7.56-7.29 (m, 6 H); 6.56 (s, 1 H); 4.69 (s, 2 H); 2.49 (s, 3 H); 1.80 (s, 1 H).

[ Manufacturing example  18] 3- (2- Hydroxymethylphenyl ) -7- methyl -2H-isoquinolin-1-one (3- (2- Hydroxymethyl - phenyl ) -7- methyl -2H- isoquinolin -One- one ) (8c) Preparation

3- (2-hydroxymethylphenyl) -7-methyl-2H-isoquinoline in the same manner as in Preparation 16 using compound 7c (1.8 g, 4.67 mmol) and DDQ (1.59 g, 7 mmol) in Preparation Example 10. Obtained 1-one ( 8c ) (yellow solid, 729 mg, 59%).

Mp: 103.7-106.0 ° C. IR (cm ⁻¹ ) :( OH), 1629 (C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 11.25 (s, 1H); 8.12 (s, 1 H); 7.55-7.36 (m, 6 H); 6.60 (s, 1 H); 5.25 (s, 1 H); 4.68 (s, 2 H); 2.50 (s, 3 H).

[ Manufacturing example  19] 3- (2- Hydroxymethyl -4- Methoxyphenyl ) -2H-isoquinolin-1-one (3- (2- Hydroxymethyl -4- methoxyphenyl ) -2H- isoquinolin -One- one ) (8d)

HCl (10 mL) was added to a mixed solution of Compound 7d (180 mg, 0.55 mmol) and THF (10 mL) in Preparation Example 11, and the mixture was refluxed for 2 hours. Water was added to the reaction mixture and extracted with EA. The organic layer was washed with water and brine, dried over anhydrous Na ₂ SO ₄ , and concentrated under reduced pressure. The residue was purified by column chromatography (CH ₂ Cl ₂ : MeOH = 20: 1) to obtain 3- (2-hydroxymethyl-4-methoxyphenyl) -2H-isoquinolin-1-one ( 8d ) ( Yellow solid, 70 mg, 45%) .

Mp: 170.6-172.3 ° C. IR (cm ⁻¹ ) :( OH), 1629 (C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 11.25 (s, 1H); 8.30 (m, 1 H); 7.64 (m, 1 H); 7.57 (m, 1 H); 7.46-7.40 (m, 2 H); 7.08 (d, J = 2.7 Hz, 1 H); 6.85 (m, 1 H); 6.57 (s, 1 H); 5.10 (s, 1 H); 4.66 (s, 2 H); 3.83 (s, 3 H).

[ Manufacturing example  20] 3- (2- Hydroxymethyl -4,5- Dimethoxyphenyl ) -2H-isoquinolin-1-one (3- (2- Hydroxymethyl -4,5- dimethoxyphenyl ) -2H- isoquinolin -One- one ) Manufacturing of (8e)

Using Compound 7e (1 g, 2.8 mmol) of Preparation Example 12 in the same manner as in Preparation Example 19, 3- (2-hydroxymethyl-4,5-dimethoxyphenyl) -2H-isoquinolin-1-one ( 8e ) (yellow solid, 200 mg, 23%).

Mp: 199.6-201.1 ° C. IR (cm ⁻¹ ) :( OH), 1629 (C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 11.55 (s, 1H); 8.29 (d, 1 H); 7.66 (m, 1 H); 7.58 (m, 1 H); 7.48 (m, 1 H); 7.10 (s, 1 H); 6.97 (s, 1 H); 6.61 (s, 1 H); 4.95 (s, 1 H); 4.64 (s, 2 H); 3.96 (s, 3 H); 3.91 (s, 3 H).

[ Manufacturing example  21] 3- (2- Hydroxymethyl -4- Methoxyphenyl ) -6- methyl -2H-isoquinolin-1-one (3- (2- Hydroxymethyl -4- methoxyphenyl ) -6- methyl -2H- isoquinolin -One- one ) (8f) Preparation

3- (2-hydroxymethyl-4-methoxyphenyl) -6-methyl-2H-isoquinoline-1- in the same manner as in Preparation Example 19 using compound 7f (892 mg, 2.6 mmol) in Preparation Example 13 Warm ( 8f ) was obtained (yellow solid, 230 mg, 30%).

Mp: 171.8-173.9 ° C. IR (cm- ¹ ): 3219 (OH), 1629 (C = O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 11.59 (s, 1H); 8.15 (d, 1 H); 7.35 (m, 2 H); 7.26 (d, 1 H); 7.11 (d, 1 H); 6.83 (m, 1 H); 6.48 (s, 1 H); 5.50 (s, 1 H); 4.64 (s, 2 H); 3.81 (s, 3 H); 2.47 (s, 3 H).

[ Manufacturing example  22] 3- (2- Hydroxymethyl -4,5- Dimethoxyphenyl ) -6- methyl -2H-isoquinolin-1-one (3- (2- Hydroxymethyl -4,5- dimethoxyphenyl ) -6- methyl -2H- isoquinolin -One- one ) (8g)

3- (2-hydroxymethyl-4,5-dimethoxyphenyl) -6-methyl-2H-isoquinoline- in the same manner as in Preparation Example 19 using 7g (1.2 g, 3.2 mmol) of the compound of Preparation Example 14 1-one ( 8 g ) was obtained (yellow solid, 120 mg, 12%).

Mp: 208.1-209.2 ° C. IR (cm- ¹ ): 219 (OH), 1629 (C = O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 11.30 (s, 1H); 8.19 (d, 1 H); 7.37 (s, 2 H); 7.26 (d, 1 H); 7.10 (s, 1 H); 6.96 (s, 1 H); 6.54 (s, 1 H); 4.64 (s, 2 H); 4.40 (s, 1 H); 3.97-3.91 (d, 6 H); 2.49 (s, 3 H).

[ Manufacturing example  23] 3- (2- Hydroxymethyl -4- Methoxyphenyl ) -7- methyl Preparation of -2H-isoquinolin-1-one (3- (2-Hydroxymethyl-4-methoxyphenyl) -7-methyl-2H-isoquinolin-1-one) (8h)

3- (2-hydroxymethyl-4-methoxyphenyl) -7-methyl-2H-isoquinoline-1- in the same manner as in Preparation Example 19 using compound 7h (520 mg, 1.5 mmol) in Preparation Example 15 Warm ( 8 h ) was obtained (yellow solid, 230 mg, 50%).

Mp: 193.3-195.5 ° C. IR (cm- ¹ ): 3219 (OH), 1629 (C = O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 11.30 (s, 1H); 8.08 (s, 1 H); 7.46 (m, 2 H); 7.38 (d, 1 H); 7.06 (d, 1 H); 6.79 (m, 1 H); 6.53 (s, 1 H); 5.45 (s, 1 H); 4.65 (s, 2 H); 3.79 (s, 3 H); 2.49 (s, 3 H).

[ Manufacturing example  24] 3- (2- Hydroxymethyl -4,5- Dimethoxyphenyl ) -7- methyl -2H-isoquinolin-1-one (3- (2- Hydroxymethyl -4,5- dimethoxyphenyl ) -7- methyl -2H- isoquinolin -One- one ) (8i)

3- (2-hydroxymethyl-4,5-dimethoxyphenyl) -7-methyl-2H-isoquinoline- in the same manner as in Preparation Example 19 using compound 7i (630 mg, 1.7 mmol) of Preparation Example 16 1-one ( 8i ) was obtained (yellow solid, 60 mg, 11%).

Mp: 114-115.2 ° C. IR (cm- ¹ ): 9 (OH), 1629 (C = O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 8.11 (s, 1H); 7.50 (s, 2 H); 7.07 (s, 1 H); 6.97 (s, 1 H); 6.58 (s, 1 H); 4.64 (s, 2 H); 3.96-3.91 (d, 6H); 2.50 (s, 3 H).

[ Example  1] 7H- Isoindolo [2,1-b] isoquinoline -5-one (7H- Isoindolo Preparation of [2,1-b] isoquinolin-5-one) (9a)

Trimethylamine (46 mg, 0.4 mmol) and methanesulfonylchloride (101 mg, 1.0 mmol) were added to a mixed solution of Compound 8a (50 mg, 0.2 mmol) and CH ₂ Cl ₂ of Preparation Example 16 at 0 ° C. The reaction mixture was allowed to warm to room temperature and stirred overnight. The reaction was quenched with water and extracted with CH ₂ Cl ₂ . The organic layer was washed with water and brine, dried over anhydrous Na ₂ SO ₄ , and concentrated under reduced pressure. The residue was purified by column chromatography ( n -hexane: EA = 3: 1) to obtain the title compound 7H-isoindolo [2,1-b] isoquinolin-5-one ( 9a ) (yellow solid, 24 mg). , 51%).

Mp: 151.7-154.1 ° C. IR (cm ⁻¹ ) :( C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 8.48 (d, J = 8.1 Hz, 1H); 7.75-7.43 (m, 7 H); 6.98 (s, 1 H); 5.15 (s, 2 H).

[ Example  2] 2- methyl -7H- Isoindolo [2,1-b] isoquinoline -5-one (2- Methyl -7H-isoindolo [2,1-b] isoquinolin-5-one) (9b)

Using the compound 8b (270 mg, 1 mmol), triethylamine (506 mg, 5 mmol) and methanesulfonylchloride (229 mg, 2 mmol) in Preparation Example 17, the target compound 2-methyl was used in the same manner as in Example 1. -7H-isoindolo [2,1-b] isoquinolin-5-one ( 9b ) was obtained (yellow solid, 94 mg, 38%).

Mp: 157.8-160.2 ° C. IR (cm ⁻¹ ) :( C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 8.38 (d, J = 8.1 Hz, 1H); 7.80 (m, 1 H), 7.57 (m, 1 H); 7.48 (m, 2 H); 7.42 (s, 1 H); 7.31 (d, 1 H); 6.97 (s, 1 H); 5.19 (s, 2 H); 2.50 (s, 3 H).

[ Example  3] 3- methyl -7H- Isoindolo [2,1-b] isoquinoline -5-one (3- Methyl -7H-isoindolo [2,1-b] isoquinolin-5-one) (9c)

Compound 8c (300 mg, 1.1 mmol), triethylamine (572 mg, 5.6 mmol) and methanesulfonylchloride (259 mg, 2.3 mmol) of Preparation Example 18 were prepared in the same manner as in Example 1, target compound 3- Methyl-7H-isoindolo [2,1-b] isoquinolin-5-one ( 9c ) was obtained (yellow solid, 111 mg, 40%).

Mp: 155.2-158.8 ° C. IR (cm ⁻¹ ) :( C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 8.30 (s, 1H); 7.79 (m, 1 H); 7.57-7.45 (m, 5 H); 7.02 (s, 1 H); 5.20 (s, 2 H); 2.51 (s, 3 H).

[ Example  4] 9- Methoxy -7H- Isoindolo [2,1-b] isoquinoline -5-on (9- Methoxy -7H-isoindolo [2,1-b] isoquinolin-5-one) (9d)

Using the compound 8d (65 mg, 0.23 mmol), triethylamine (65 mg, 0.23 mmol) and methanesulfonylchloride (58 mg, 0.5 mmol) of Preparation Example 19, the desired compound 9- Methoxy-7H-isoindolo [2,1-b] isoquinolin-5-one ( 9d ) was obtained (yellow solid, 35 mg, 58%).

Mp: 163.8-165.3 ° C. IR (cm- ¹ ): 1657 (C = O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 8.46 (d, 1H); 7.70-7.59 (m, 3 H); 7.43 (m, 1 H); 7.05-7.00 (m, 2 H); 6.88 (s, 1 H); 5.13 (s, 2 H); 3.88 (s, 3 H).

[ Example  5] 9,10- Dimethoxy -7H- Isoindolo [2,1-b] isoquinoline Preparation of -5-one (9,10-Dimethoxy-7H-isoindolo [2,1-b] isoquinolin-5-one) (9e)

Using compound 8e (200 mg, 0.64 mmol), triethylamine (338 mg, 3.3 mmol) and methanesulfonylchloride (153 mg, 1.3 mmol) in Preparation Example 20, the desired compound 9, 10-dimethoxy-7H-isoindolo [2,1-b] isoquinolin-5-one ( 9e ) was obtained (yellow solid, 66 mg, 35%).

Mp: 226.4-228.0 ° C. IR (cm ⁻¹ ) :( C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 8.46 (d, 1H); 7.63-7.60 (m, 2 H); 7.45 (m, 1 H); 7.21 (s, 1 H); 7.03 (s, 1 H); 6.88 (s, 1 H); 5.10 (s, 2 H); 3.99 (d, 6 H).

[ Example  6] 9- Methoxy -2- methyl -7H- Isoindolo [2,1-b] isoquinoline -5-on (9- Methoxy -2- methyl -7H- isoindolo [2,1-b] isoquinolin-5- one ) Production of (9f)

Target compound 9 in the same manner as in Example 1 using compound 8f (200 mg, 0.68 mmol), triethylamine (344 mg, 3.4 mmol) and methanesulfonylchloride ((155 mg, 1.36 mmol) in Preparation Example 21 -Methoxy-2-methyl-7H-isoindolo [2,1-b] isoquinolin-5-one ( 9f) was obtained (yellow solid, 60 mg, 32%).

Mp: 188.6-190.8 ° C. IR (cm ⁻¹ ) :( C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 8.34 (d, 1H); 7.67 (d, 1 H); 7.36 (s, 1 H); 7.26 (m, 1 H); 7.05-6.99 (m, 2 H); 6.81 (s, 1 H); 5.12 (s, 2 H); 3.88 (s, 3 H); 2.48 (s, 3 H).

[ Example  6] 9,10- Dimethoxy -2- methyl -7H- Isoindolo [2,1-b] isoquinoline -5-one (9,10- Dimethoxy -2- methyl -7H- isoindolo [2,1-b] isoquinolin-5- one ) (9g)

Using the compound 8g (100 mg, 0.31 mmol), triethylamine (161 mg, 1.6 mmol) and methanesulfonylchloride (74 mg, 0.64 mmol) in Preparation Example 22, the desired compound 9, 10-dimethoxy-2-methyl-7H-isoindolo [2,1-b] isoquinolin-5-one ( 9 g) was obtained (yellow solid, 50 mg, 52%).

Mp: 235-237 ° C. IR (cm ⁻¹ ) :( C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 8.35 (d, 1H); 7.36 (s, 1 H); 7.25 (m, 1 H); 7.20 (s, 1 H); 7.03 (s, 1 H); 6.80 (s, 1 H); 5.09 (s, 2 H); 3.99 (d, 6 H); 2.49 (s, 3 H).

[ Example  7] 9- Methoxy -3- methyl -7H- Isoindolo [2,1-b] isoquinoline -5-on (9- Methoxy -3- methyl -7H- isoindolo [2,1-b] isoquinolin-5- one ) Production of (9h)

Using the compound 8h (100 mg, 0.34 mmol), triethylamine (344 mg, 3.4 mmol) and methanesulfonylchloride (78 mg, 0.68 mmol) in Preparation Example 23, the target compound 9- was used in the same manner as in Example 1. Methoxy-3-methyl-7H-isoindolo [2,1-b] isoquinolin-5-one ( 9h) was obtained (yellow solid, 46 mg, 49%).

Mp: 180.4-182.9 ° C. IR (cm ⁻¹ ) :( C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 8.21 (s, 1H); 7.62 (d, 1 H); 7.47-7.42 (m, 2 H); 6.99-6.94 (m, 2H); 6.79 (s, 1 H); 5.06 (s, 2 H); 3.86 (s, 3 H); 2.46 (s, 3 H).

[ Example  8] 9,10- Dimethoxy -3- methyl -7H- Isoindolo [2,1-b] isoquinoline Preparation of -5-one (9,10-Dimethoxy-3-methyl-7H-isoindolo [2,1-b] isoquinolin-5-one) (9i)

Using compound 8i (50 mg, 0.15 mmol), triethylamine (86 mg, 0.85 mmol) and methanesulfonylchloride (38 mg, 0.34 mmol) in Preparation Example 24, the desired compound 9, 10-dimethoxy-3-methyl-7H-isoindolo [2,1-b] isoquinolin-5-one ( 9i) was obtained (brown solid, 17 mg, 36%).

Mp: 208.6-210.5 ° C. IR (cm ⁻¹ ) :( C═O). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 8.27 (s, 1H); 7.49 (m, 2 H); 7.21 (s, 1 H); 7.04 (s, 1 H); 6.87 (s, 1 H); 5.11 (s, 2 H); 3.98 (d, 6 H); 2.50 (s, 3 H).

[ Experimental Example  1] according to the present invention Isoindolo [2,1-b] isoquinolinone  Derivative On cancer cell lines  Against cytotoxicity and Topoisomerase  I inhibitory activity assay

The anticancer activity of the isoindolo [2,1-b] isoquinolinone derivatives of the general formula (1) according to the present invention is known as the method of Sulforhodamine B (SRB, Sulforhodamine B) as described below [P. Skehan, et. al., New colorimetric cytotoxicity assay for anti-cancer drug screening, J. Natl. Cancer Inst., 1990, 82, 1107-1112] was measured by in vitro experiments. The test cancer cell lines A549 (human lung cancer cell line), HCT15 (human colorectal cancer cell line), and OV-3 (human ovarian cancer cell line) were used in this experiment.

Each cancer cell line was incubated for 48 hours in an incubator at 37 ° C. in the presence of 5% CO ₂ using Sigma's RPMI1640 medium, and dissolved in dimethyl sulfoxide (DMSO) at a concentration of 1 mg / ml. The solution was diluted in 10-fold dilutions and added, followed by further incubation for 48 hours, after which the culture medium was removed. 50% trichloroacetic acid (TCA) was added thereto at a concentration of 50 μl per well, and the cells were fixed for 1 hour (final concentration 10%). After TCA was removed and the plate was washed with distilled water and dried, 50 μl of 0.4% sulforhodamine B (SRB) dissolved in 1% acetic acid as a dye was added per well. After 20 minutes sulforhodamine B was removed and washed four times with 1% acetic acid solution and then dried. 150 μl of 10 mmol / L unbuffered trisbase {tris (hydroxymethyl) aminomethane} was added to each well, and the mixture was mixed well and absorbance was measured at 540 nm. From the measured absorbance, the concentration of a drug that inhibits the proliferation of cancer cells by 50% using a microplate reader was calculated as anticancer activity (ED ₅₀ ).

The topoisomerase I inhibitory activity was determined by measuring the relaxed form of supercoil pBR322 plasmid DNA using calf thymus DNA topoisomerase I. Relaxing buffer mixture of a compound prepared in Example (20 mM in DMSO) or a comparative material camptothecin (20 mM in DMSO), 200 mg of plasmid pBR322 DNA and 2 units of calf thymus DNA topoisomerase I (Fermentas, USA) The solution [35 mM tris-HCl (pH 8.0), 72 mM KCl, 5 mM MgCl ₂ , 5 mM dithiopretol, 2 mM spermidine, 0.01% bovine serum albumin] was incubated for 30 minutes at 37 ℃. The reaction (last volume 20 L) was purified by adding 5 L of stop buffer containing 10% SDS, 0.2% Bromophenol blue, 0.2% Xylene cyanol and 30% glycerol. And loaded onto 1% agarose gel and developed in TAE (Tris-Acetate-EDTA) buffer. Electrophoresis was performed slowly at 15V for 6 hours, and the developed gel was dyed for 30 minutes with an aqueous solution of ethidium bromide (EtBr) (0.5 ug / ml). After separation, the gel was exposed to UV light and DNA bands were quantified using AlphaImager ^™ .

The results of cytotoxicity and topoisomerase I inhibition of the A549 (human lung cancer cell line), HCT15 (human colon cancer cell line) and OV-3 (human ovarian cancer cell line) are shown in Table 1 and FIG. 1.

TABLE 1

As shown in Table 1, the isoindolo [2,1-b] isoquinolinone derivatives according to the present invention not only have excellent cytotoxicity against human cancer cells but also have an inhibitory effect on topoisomerase I. And it was found. Further, from FIG. 1, the compounds of Examples 1 to 7 <lane E ( 9a ), lane F ( 9b ), lane G ( 9c ), lane H ( 9d ), lane I ( 9f ), lane J ( 9e ) , Lane K ( 9g )> inhibited topoisomerase I in vitro. In particular, compounds 9a and 9d of Example 1 and Example 4 had similar inhibitory activity as camptothecin. Therefore, the isoindolo [2,1-b] isoquinolinone derivative according to the present invention has an excellent inhibitory effect on topoisomerase, and also has excellent cytotoxicity against human cancer cells, and thus can be usefully used as a cancer-containing agent.

1-Topoisomerase I inhibitory activity of isoindolo [2,1-b] isoquinolinone derivatives according to the present invention [lane A and L: pBR322 alone, lane B: pBR322 + topoisomerase I, lane C: pBR322 + topoisomerase I + camptothecin (100 mg / mL), lane D: pBR322 + topoisomerase I + camptothecin (100 mg / mL), lane EK, isoindolo according to the invention [ 2,1-b] isoquinolinone derivatives <lane E ( 9a ), lane F ( 9b ), lane G ( 9c ), lane H ( 9d ), lane I ( 9f ), lane J ( 9e ), lane K ( 9 g )> (100 mg / mL) + pBR322 + topoisomerase I]

Claims (9)

Isoindolo [2,1-b] isoquinolinone derivative represented by the following formula (1). [Formula 1]

[In Formula 1, R ₁ to R ₄ are independently of each other hydrogen, halogen or (C1-C7) alkyl, and R ₁₁ to R ₁₄ are independently of each other hydrogen, halogen or (C1-C7) alkoxy.] The method of claim 1, Isoindolo [2,1-b], wherein R ₁ and R ₂ are independently of each other hydrogen, chloro, fluorine or methyl, and R ₁₁ and R ₁₂ are independently of each other hydrogen, chloro, fluorine or methoxy. Isoquinolinone derivatives. 3. The method of claim 2, Isoindolo [2,1-b] isoquinolinone derivatives, characterized in that selected from the following structures:

A method for preparing an isoindolo [2,1-b] isoquinoline derivative of Chemical Formula 1 by intramolecular ring addition reaction of a hydroxymethyl compound of Chemical Formula 2 [Formula 1]

[Formula 2]

[In Formulas 1 and 2, R ₁ to R ₄ are independently of each other hydrogen, halogen or (C1-C7) alkyl, and R ₁₁ to R ₁₄ are independently of each other hydrogen, halogen or (C1-C7) alkoxy. ] The method of claim 4, wherein The hydroxymethyl compound of Formula 2 is prepared by reacting the compound of Formula 3 with DDQ (2,3-dichloro-5,6-dicyano-l, 4-benzoquinone). [Formula 2]

(3)

[In the above formula, R ₁ to R ₄ are independently of each other hydrogen, halogen or (C1-C7) alkyl, R ₁₁ to R ₁₄ are independently of each other hydrogen, halogen or (C1-C7) alkoxy, R ₁₅ is (C1-C7) alkoxy (C1-C7) alkyl or (C1-C7) alkoxy (C6-C12) aryl.] The method of claim 5, The compound of Formula 3 is prepared by reacting an o-toluimide compound of Formula 4 with a benzonitrile compound of Formula 5. (3)

[Formula 4]

[Chemical Formula 5]

[In the above formula, R ₁ to R ₄ are independently of each other hydrogen, halogen or (C1-C7) alkyl, R ₅ and R ₆ are independently of each other hydrogen or (C1-C7) alkyl, R ₁₁ to R ₁₄ Are independently of each other hydrogen, halogen or (C1-C7) alkoxy, and R ₁₅ is (C1-C7) alkoxy (C1-C7) alkyl or (C1-C7) alkoxy (C6-C12) aryl.] An anticancer agent composition comprising the isoindolo [2,1-b] isoquinolinone derivative according to any one of claims 1 to 3 as an active ingredient. The method of claim 7, wherein An anticancer composition characterized by inhibiting the activity of topoisomerase I. The method of claim 7, wherein Anticancer composition, characterized in that the cancer is lung cancer, colorectal cancer or ovarian cancer.

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