KR100201060B1 - Benzindole derivatives - Google Patents

Abstract

The present invention relates to a novel benzindole derivative of the following general formula (I) and a pharmaceutically acceptable salt thereof, a method for preparing the same, and an anticancer composition containing the compound as an active ingredient, which inhibits the proliferation of cancer cells.

In the above formula,

R represents a group of -CO-R ₁ , -CONH-R ₂ or -SO ₂ -NH-R ₂ , wherein R ₁ is unsubstituted or substituted by alkyl and is at least one selected from nitrogen, sulfur and oxygen Mono- or di-heterocyclic group containing a hetero atom,

의 그룹(여기에서 R₃는 아릴을 나타내고, R₄는 카르복실, 아미노카르보닐 또는 알콕시카르보닐을 나타낸다) 또는 아릴술포닐을 나타낸다.R ₂ is aryl unsubstituted or substituted by hydroxycarbonyl, aminocarbonyl or alkoxycarbonyl,

R ₃ represents aryl and R ₄ represents carboxyl, aminocarbonyl or alkoxycarbonyl or arylsulfonyl.

Description

Benzindole derivatives

The present invention relates to a novel benzindole derivative of the following general formula (I) and a pharmaceutically acceptable salt thereof, a preparation method thereof, and an anticancer composition containing the compound as an active ingredient, which inhibits the proliferation of cancer cells.

In the above formula,

R represents a group of -CO-R ₁ , -CONH-R ₂ or -SO ₂ -NH-R ₂ , wherein R ₁ is unsubstituted or substituted by alkyl and is at least one selected from nitrogen, sulfur and oxygen Mono- or di-heterocyclic group containing a hetero atom,

의 그룹(여기에서 R₃는 아릴을 나타내고, R₄는 카르복실, 아미노카르보닐 또는 알콕시카르보닐을 나타낸다) 또는 아릴술포닐을 나타낸다.R ₂ is aryl unsubstituted or substituted by hydroxycarbonyl, aminocarbonyl or alkoxycarbonyl,

R ₃ represents aryl and R ₄ represents carboxyl, aminocarbonyl or alkoxycarbonyl or arylsulfonyl.

Until now, various compounds having various cancer cell proliferation inhibitory activities have been developed, but it has not been found that compounds having a structure similar to that of the general formula (I) have a cancer cell proliferation inhibitory effect useful as an anticancer agent. ) Is a useful cancer cell proliferation inhibitor with a new structure.

Preferred compounds of the general formula (I) according to the invention are those in which R represents a group of the structure:

Particularly preferred compounds of general formula (I) are those wherein R represents a group of the structure:

Asymmetric carbon atoms may be included in the structure of the compounds according to the invention, and therefore the compounds of general formula (I) may exist in the form of racemates or individual optically active isomers. Such isomers are included in the scope of the compounds of the general formula (I) according to the present invention.

Compounds of general formula (I) according to the invention may form salts with pharmaceutically acceptable non-toxic acids, which salts are also within the scope of the compounds of the invention. Examples of such acid addition salts include organic acids such as acetic acid, trifluoroacetic acid, maleic acid, fumaric acid, lactic acid, oxalic acid, tartaric acid, citric acid, glucuronic acid, methanesulfonic acid, benzenesulfonic acid and various amino acids, and sulfuric acid, hydrochloric acid, hydrobromic acid, nitric acid, and the like. And salts formed by inorganic acids such as phosphoric acid and the like.

The invention also relates to a process for the preparation of the novel benzindole derivatives of the general formula (I). According to the present invention, the following compound of general formula (II) is alkylated with the compound of general formula (III), and then alkylated with the compound of general formula (IV), and Lawson reagent (2,4) -Bis (4-methoxyphenyl) -1,3-dithia-2,4-diphosphane-2,4-disulfide) was used to prepare the compound of formula (V): The desired benzindole derivative of the general formula (I) can be prepared by reacting the compound of formula (V) with the compound of the general formula (VI) to produce methylthio compound (VI), followed by ammonia substitution.

In the above formula

R 'has the same meaning as R as defined above, or represents an alkyl-substituted ester or sulfonylamide, provided that when amide groups are present in R', the amide nitrogen is subjected to hydrogenation in the presence of a metal catalyst. T-butoxycarbonyl, tetrabutylammonium fluoride which can be removed by hydrolysis with acids such as benzyl, benzyloxy, carbenzyloxy, benzyloxymethyl, etc., which may be removed, hydrochloric acid, trifluoroacetic acid Appropriate protecting groups such as t-butyldimethylsilyloxymethyl group, which can be removed by, may be introduced, X is a halogen atom such as iodine, bromine, chlorine or the like or a suitable reactive leaving such as alkylsulfonyloxy, arylsulfonyloxy and the like Group.

According to the process of the invention, the compound of general formula (II) is first subjected to a two-step alkylation reaction. In the above process, the two-step alkylation reaction may be carried out by changing the reaction order as necessary.

The two step alkylation can be carried out in the presence of a base in a reaction-inert solvent. Examples of reaction-inert solvents that can be preferably used for this purpose include dimethylformamide, dimethylacetamide, diketylsulfoxide, tetrahydrofuran, dioxane and the like, and preferred bases are trimethylamine, triethylamine, di Organic bases such as isopropylethylamine, N, N-dimethylaniline and the like, or inorganic bases such as sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide and the like can be used. The reaction temperature is not particularly limited, but in general, the reaction is carried out with stirring for 1 hour to 2 days at a temperature between room temperature and the boiling point of the solvent used.

The compound produced by the two-step alkylation of the compound of formula (II) is subsequently reacted with Lawson's reagent to replace oxygen at the lactam site with sulfur to give the compound of formula (V). The reaction with Lawson reagent can preferably be carried out in a reaction-inert solvent. Examples of reaction-inert solvents that can be preferably used for this purpose include benzene, toluene, xylene, nitrobenzene, chlorobenzene and the like.

The reaction is generally carried out at a temperature between room temperature and the boiling point of the solvent used, preferably under reflux for 1 hour to 2 days.

The compound of formula (V) produced by reaction with Lawson reagent is again alkylated with a compound of formula (IV) to give a compound of formula (VI). This reaction can be carried out under the same conditions as the aforementioned alkylation reaction conditions.

The produced compound of general formula (VI) can finally produce the compound of general formula (I) by carrying out ammonia substitution. This ammonia substitution reaction can be carried out by saturating ammonia in an organic solvent which does not adversely affect the reaction. Examples of the solvent preferably used for this purpose include lower alkanols such as methanol, ethanol, or tetrahydrofuran, dioxane. , Acetone and the like. The reaction temperature and pressure are not particularly limited, and generally, the reaction is carried out for 1 hour to 2 days under normal pressure to a pressure in a sealed container at a temperature different from the boiling point of the solvent.

The resulting compound of formula (I) according to the present invention is reacted with a suitable organic or inorganic acid capable of forming acid addition leaves as mentioned above according to conventional salt formation methods, if necessary, of compounds of general formula (I). Can form acid addition salts.

Benzindole derivatives of the general formula (I) and salts thereof produced by the method of the present invention can be separated and purified by conventional post-treatment methods, for example, by recrystallization, column chromatography, and the like.

The novel benzindole derivatives of general formula (I) according to the present invention can be usefully used as therapeutic agents for various diseases caused by abnormal cell proliferation, in particular as anticancer agents, by inhibiting DNA synthesis by inhibiting DNA synthesis as folate antagonists. . For example, the cell proliferation inhibitory effect of the compound of general formula (I) may be expressed in the form of antibiotics, antifungal action, antiviral action, psoriasis suppression as well as anticancer action.

Therefore, another object of the present invention is to provide a cancer-containing composition containing the compound of the general formula (I) as an active ingredient.

The composition according to the present invention can be combined with a pharmaceutically acceptable liquid or solid carrier for clinical use to prepare a conventional formulation, such as tablets (sugar drags, divided tablets, etc.), according to pharmaceutically conventional methods. Oral preparations, such as capsules, troches, solutions, suspensions, or injectable solutions or suspensions, or ready-to-use injectable dry powders that can be reconstituted with injectable distilled water at the time of injection. It may be formulated into a variety of preparations, such as injectable preparations, to be administered orally or parenterally, for example, intramuscularly, intravenously or intraarterally. In addition, in order to prevent the decomposition of the drug by gastric acid during oral administration, an antacid may be used in combination, or a solid dosage form for oral administration such as tablets may be formulated into a formulation coated with enteric skin.

Liquid carriers used in the formulation of the composition of the present invention include water, saline, olive oil, syrup, etc., and solid carriers include starch, agar, gelatin, pectin, sucrose, lactose, calcium sulfate, stearic acid and magnesium stearate. Etc.

The dosage of the compound of general formula (I) according to the present invention to the human body is determined by the absorption rate, inactivation rate and excretion rate of the active ingredient in the body, the age, sex, weight and general physical condition of the patient, the type and severity of the disease to be treated, etc. It is appropriately selected according to the above, but generally, an amount of 10 to 2000 mg is administered at a time and is administered in 1 to 3 weeks.

The invention is illustrated in more detail by the following examples. However, the following examples are provided merely to illustrate the present invention and the present invention is not limited in any way by these.

The structures of the compounds prepared in the following examples were determined by nuclear magnetic resonance spectroscopy, infrared spectroscopy, mass spectrometry, and elemental microanalysis.

Quantum nuclear magnetic resonance spectra were measured on a Bruker AMX-R300 (300 MHz), and chemical shifts were expressed as ppm downfield shifts as δ scale, based on the internal standard tetramethylsilane, with overlapping peaks. Is denoted as follows: s, singlet; d, doublet; dd, doublet of doublets; t, triplet; q, quartet; m, multiplet; br, broad.

Infrared spectroscopy spectra were measured in the form of KBr pellets on Perkin-Elmer FTIR 1750, and mass spectrometry spectra were measured on an Extrell ELQ-400 using ammonia as a mobile gas. Elemental microanalysis was performed using CE instrument EA1110, which was used as the reference within ± 0.4% of theory for each element.

Melting points were measured with a Thomas Hoover capillary melting point meter and were not corrected. Thin layer chromatography (TLC) using a Merck silica gel 60 F-254 coated glass plate, ultraviolet rays, iodine, phosphomolybdic acid, anisealdehyde was observed by color development, and flash column chromatography was performed on Merck silica gel 60 (40-63). Micrometers).

Tetrahydrofuran and dioxane in the solvent used for the reaction were dried by distillation with sodium benzophenone ketil, dimethylformamide was dried by distillation with magnesium sulfate, and methylene chloride was dried with calcium hydride and distilled.

Example 1

: Preparation of Starting Material (II)

1) Preparation of 6-nitrobenz [cd] indole-2 (1H) -one (Compound 1)

10.0 g (59.0 mmol) of benz [cd] indole-2 (1H) -one (Aldrich 33,646-7) was dissolved in 50 mL of glacial acetic acid, and 3.75 mL (76.7 mmol) of 86% nitric acid was added dropwise. The reaction solution was stirred for 1 hour, and then added to 500 mL of ice-water to give a yellow solid. The resulting solid was filtered and washed with water until the yellow no longer melted and dried. After dissolving this solid in tetrahydrofuran, the insoluble solid was filtered off and the filtrate was concentrated under reduced pressure. 8.0 g of the resulting solid was again dissolved in 150 ml of methanol, cooled to 0 ° C., and filtered to obtain 6.7 g (yield 53%) of a yellow solid (Compound 1).

Melting Point: 298.0 ~ 300.0 ℃

IR (KBr): 3316, 1724, 1638, 1491, 1351, 1252, 1051, 752 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 7.14 (d, 1H, J = 8.0 Hz), 8.09 (dd, 1H, J = 7.1 Hz), 8.19 (d, 1H, J = 7.0 Hz), 8.65 (d , 1H, J = 7.1 Hz), 8.90 (d, 1H, J = 8.4 Hz)

2) Preparation of 6-aminobenz [cd] indole-2 (1H) -one (Compound II)

6.5 g (30.3 mmol) of 6-nitrobenz [cd] indole-2 (1H) -one was dissolved in 200 ml of tetrahydrofuran, 0.7 g of 5% palladium-activated carbon was added to the solution, followed by Parr hydrogenator. ) Under a pressure of 40 psi for 16 hours. After the reaction was completed, the mixture was filtered using Celite and the filtrate was concentrated under reduced pressure. 5.5 g of the resulting solid was dissolved in 30 ml of tetrahydrofuran again, and 200 ml of n-hexane was added to crystallize, followed by filtration to obtain 5.1 g (yield 91%) of a red solid (Compound II).

Melting Point: 240.0 ~ 242.0 ℃

IR (KBr): 3490.3287, 2905, 1720, 1672, 1647, 1477, 777cm ^-1

¹ H NMR (acetone-d ₆ ): δ 5.07 (br, 2H), 6.51 (d, 1H, J = 7.5 Hz), 6.66 (d, 1H, J = 7.5 Hz), 7.58 (dd, 1H, J = 7.0 Hz), 7.85 (d, 1H, J = 7.0 Hz), 8.21 (d, 1H, J = 7.1 Hz), 9.22 (br, 1H)

Example 2

: N ⁶ - Preparation of ^6-methyl -N 8)

1) Preparation of N ^6- (4-methoxycarbonylbenzyl) -6-aminobenz [cd] indole-2 (1H) -one (Compound 2)

3.0 g (16.29 mmol) of 6-aminobenz [cd] indole-2 (1H) -one and 6.24 mL (35.83 mmol) of diisopropylethylamine were dissolved in 100 mL of dimethylformamide and 4- (bromomethyl) benzoic acid 3.73 g (16.29 mmol) of methyl ester (Aldrich 34,815-5) were added and reacted at room temperature for 6 hours. After the reaction, the mixture was added to 800 ml of water and filtered when a solid formed. This solid was dissolved in methylene chloride, dried over sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by flash column chromatography using a 1: 1 mixture of methylene chloride and ethyl acetate to give 3.73 g (yield 69%) of an orange solid (Compound 2).

¹ H NMR (DMSO-d ₆ ): δ 4.57 (d, 2H, J = 5.5 Hz), 6.12 (d, 1H, J = 7.6 Hz), 6.69 (d, 1H, J = 7.6 Hz), 7.70 (d , 2H, J = 8.3 Hz, 7.80 (t, 1H, J = 7.0 Hz), 7.92 to 8.00 (m, 3H), 8.15 (d, 1H, J = 8.2 Hz), 10.40 (s, 1H)

2) N ⁶ - Preparation of one (Compound 3) -methyl -N ⁶ - (4- methoxycarbonyl-benzyl) -6-amino benz [cd] indol -2 (1H)

3.0 g (9.03 mmol) of N ^6- (4-methoxycarbonylbenzyl) -6-aminobenz [cd] indol-2 (1H) -one and 2.36 mL (13.54 mmol) of diisopropylethylamine were added to dimethylformamide. It was dissolved in 30 ml, 0.67 ml (10.83 mmol) of iodomethane was added and stirred overnight at room temperature.

After the reaction, the mixture was added to 750 ml of water and filtered when an orange solid formed. This solid was dissolved in methylene chloride, dried over sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by flash column chromatography using a 1: 1 mixture of methylene chloride and ethyl acetate to give 2.0 g (yield 65%) of an orange solid (Compound 3).

Melting Point: 162.4 ~ 163.1 ℃

IR (KBr): 3200, 2950, 1730, 1700, 1480, 1290, 1130 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 2.78 (s, 3H), 3.85 (s, 3H), 4.42 (s, 2H), 6.85 (d, 1H, J = 7.5Hz), 6.97 (d, 1H, J = 7.6 Hz), 7.57 (d, 2H, J = 8.3 Hz), 7.79 (t, 1H, J = 7.1 Hz), 7.94 to 8.01 (m, 3H), 8.29 (d, 1H J = 8.2 Hz), 10.60 (s, 1H)

Mass: m / z 347 (M ⁺ )

3) N ⁶ - Preparation of thione (Compound 4) -methyl -N ⁶ - (4- methoxycarbonyl-benzyl) -6-amino benz [cd] indol -2 (1H)

1.8 g (5.20 mmol) of N ⁶ -methyl-N ^6- (4-methoxycarbonylbenzyl) -6-aminobenz [cd] indole-2 (1H) -one were mixed with 70 ml of toluene and stirred under reflux. 2.31 g (5.72 mmol) of Lawson's reagent were added and the mixture was stirred under reflux for 1 hour. The reaction mixture was concentrated under reduced pressure to remove the solvent, and purified by flash column chromatography using a 1: 1 mixture of methylene chloride and ethyl acetate to obtain 1.33 g (yield 71%) of a red solid (Compound 4).

IR (KBr): 3450, 3200, 2950, 1730, 1450, 1300, 1210 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 2.86 (s, 3H), 3.85 (s, 3H), 4.54 (s, 2H), 6.95 (d, 1H, J = 7.7Hz), 7.08 (d, 1H, J = 7.7 Hz), 7.55 (d, 2H, J = 8.2 Hz), 7.78 (t, 1H, J = 8.1 Hz), 7.98 (d, 2H, J = 8.2 Hz), 8.14 (d, 1H J = 7.1 Hz), 8.34 (d, 1H J = 8.2 Hz), 12.61 (s, 1H)

Mass: m / z 363 (M ⁺ )

4) Preparation of N-methyl-N- (4-methoxycarbonylbenzyl) -2-methylthio-6-aminobenz [cd] indole (Compound 5)

N ⁶ -methyl-N ^6- (4-methoxycarbonylbenzyl) -6-aminobenz [cd] indole-2 (1H) -thione 1.3 g (3.59 mmol) in ethanol and tetrahydrofuran 1: 1 mixture 10 After dissolving in mL, 7.8 mL (7.8 mmol) of 1N aqueous sodium hydroxide solution was added dropwise at room temperature, followed by adding 0.25 mL (3.95 mmol) of iodine methane and stirring for 30 minutes. 50 ml of water was added thereto, extracted three times with 70 ml of ethyl acetate, washed with brine and dried over sodium sulfate. The residue obtained by concentrating the organic solution under reduced pressure was purified by means of plate column chromatography using a 7: 1 mixture of methylene chloride and ethyl acetate to obtain 1.0 g (yield 75%) of a red solid (Compound 5).

Melting Point: 98.7 ~ 100.0 ℃

IR (KBr): 3050, 2950, 1730, 1450, 1280, 940 cm ^-1

¹ H NMR (CDCl ₃ ): δ 2.80 (s, 3H), 2.88 (s, 3H), 3.88 (s, 3H), 4.57 (s, 2H), 6.81 (d, 1H, J = 7.7 Hz), 7.45 7.48 (m, 4H), 7.82 (d, 1H, J = 7.0 Hz), 8.03 (d, 2H, J = 8.2 Hz), 8.08 (d, 1H J = 8.2 Hz)

Mass: m / z 376 (M ⁺ ), 278, 227, 211

5) Preparation of N-methyl-N- (4-hydroxycarbonylbenzyl) -2-methylthio-6-aminobenz [cd] indole (Compound 6)

0.95 g (2.52 mmol) of N-methyl-N- (4-methoxycarbonylbenzyl) -2-methylthio-6-aminobenz [cd] indole was dissolved in 12 ml of acetone and 12.6 ml (12.6 ml) of 1N aqueous lithium hydroxide solution Mmol) was added and then stirred at 35 ° C for 1 hour. After completion of the reaction, the reaction solution was concentrated under reduced pressure in half, adjusted to pH 7 with concentrated hydrochloric acid, and extracted with ethyl acetate. The organic solution was washed with brine, dried over sodium sulfate and concentrated under reduced pressure to yield 0.82 g (yield 95%) of a red solid (Compound 6).

Melting Point: 218.8 ~ 219.5 ℃

IR (KBr): 3500 (br), 2950, 2500 (br), 1720, 1470, 1250, 1140, 950, 830, 780cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 2.78 (s, 3H), 2.96 (s, 3H), 4.68 (s, 2H), 6.89 (d, 1H, J = 7.8Hz), 7.52 (d, 3H, J = 7.4 Hz), 7.67 (t, 1H, J = 7.2 Hz), 7.95 to 8.01 (m, 3H), 8.23 (d, 1H, J = 8.2 Hz)

6) Manufacture of 7)

0.80 g (2.20 mmol) N-methyl-N- (4-hydroxycarbonylbenzyl) -2-methylthio-6-aminobenz [cd] indole was dissolved in 10 ml of dimethylformamide and cooled to 0 ° C. Under argon gas, 0.58 mL (3.30 mmol) of diisopropylethylamine and 0.57 mL (2.65 mmol) of diphenylphosphoryl azide were added and stirred for 30 minutes. 0.33 g (2.20 mmol) of 3-aminobenzoic acid methyl ester (Lancaster 6907) was added, followed by stirring at room temperature for 12 hours. After the reaction, the mixture was added to 100 ml of water and filtered when a solid formed. This solid was dissolved in methylene chloride, dried over magnesium sulfate and concentrated under reduced pressure. The obtained residue was purified by column chromatography using a 1: 1 mixture of methylene chloride and ethyl acetate to give 0.62 g (yield 63%) of a red solid (Compound 7).

IR (KBr): 3450, 2950, 1740, 1605, 1450, 1300, 1280, 1050, 940, 750 cm ^-1

¹ H NMR (CDCl ₃ ): δ 2.84 (s, 6H), 3.90 (s, 3H), 4.54 (s, 2H), 6.81 (d, 1H, J = 7.0Hz), 7.27 (s, 1H), 7.37 (t, 1H, J = 7.9 Hz), 7.43 to 7.51 (m, 5H), 7.73 (s, 1H), 7.78 to 7.86 (m, 3H), 8.03 (d, 1H J = 8.2 Hz)

7) N ⁶ - methyl ^{-N 6 - [4 - [(} 3- amino-carbonyl-phenyl) aminocarbonyl] benzyl] Preparation of 2,6-diamino-benz [cd] indole (Compound 8)

0.25 g of N-methyl-N- [4-[(3-methoxycarbonylphenyl) aminocarbonyl] benzyl] -2-methylthio-6-aminobenz [cd] indole was added to 25 g of methanol solution saturated with ammonia gas. It was added to ㎖ and reacted for 5 hours at 145 ℃ in a closed vessel. After the reaction, the mixture was cooled to room temperature and the solvent was concentrated under reduced pressure. The obtained residue was purified by column chromatography using a 5: 1 mixture of methylene chloride and methanol to give 0.068 g (yield 30%) of an orange solid (Compound 8).

Melting Point: 181 ° C (Decomposition)

IR (KBr): 3350, 3200, 1670, 1610, 1450, 1220, 940, 770cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 2.70 (s, 3H), 4.37 (s, 2H), 6.79 (d, 1H, J = 7.5Hz), 6.88 (d, 1H, J = 7.5Hz), 7.25 -7.31 (m, 2H), 7.50 (s, 1H), 7.53-7.63 (m, 5H), 7.75 (d, 2H J = 8.3 Hz), 8.02 (s, 1H), 8.04-8.05 (m, 2H)

Mass: m / z 449 (M ⁺ ), 331, 285, 240, 211, 197, 179, 83

Elemental Analysis for _{C 27 H 23 N 5 O 2} · H 2 O

Calc. C 69.36; H 5.39; N 14.98

Found C 69.26; H 5.42; N 14.67

Example 3

: N ⁶ - Preparation of methyl ⁶ -N 10)

1) Manufacture of 9)

1.0 g (2.76 mmol) of N-methyl-N- (4-hydroxycarbonylbenzyl) -2-methylthio-6-aminobenz [cd] indole was dissolved in 10 ml of dimethylformamide and cooled to 0 ° C. Under argon gas, 0.72 ml (4.14 mmol) of diisopropylethylamine and 0.65 ml (3.03 mmol) of diphenylphosphoryl azide were added and stirred for 30 minutes. 0.55 g (2.76 mmol) of (S)-(+)-2-phenylglycine methyl ester hydrochloride (Aldrich 30,867-6) was added, followed by stirring at room temperature for 12 hours. After the reaction, the mixture was added to 100 ml of water and filtered when a solid formed. This solid was dissolved in methylene chloride, dried over magnesium sulfate and concentrated under reduced pressure. The obtained residue was purified by column chromatography using a 1: 1 mixture of methylene chloride and ethyl acetate to give 0.80 g (yield 58%) of a red solid (Compound 9).

IR (KBr): 3450, 2950, 1750, 1650, 1500, 1450, 1240, 940 cm ^-1

¹ H NMR (CDCl ₃ ): δ 2.85 (s, 3H), 2.92 (s, 3H), 3.78 (s, 3H), 4.61 (s, 2H), 5.81 (d, 1H, J = 7.0Hz), 6.85 (d, 1H, J = 7.6 Hz), 7.27 (s, 1H), 7.37-7.49 (m, 3H), 7.50-7.55 (m, 6H), 7.84 (m, 3H), 8.11 (d, 1H, J) = 8.2 Hz)

Mass: m / z 510 (M ⁺ ), 433, 307, 229, 205, 180

Preparation of methyl ^{-N 6 10) - 2) N} 6

0.25 g was added to 25 ml of methanol solution saturated with ammonia gas and reacted at 145 ° C. for 5 hours in a sealed container. After the reaction, the mixture was cooled to room temperature and the solvent was concentrated under reduced pressure. The obtained residue was purified by column chromatography using a 5: 1 mixture of methylene chloride and methanol to give 0.10 g (yield 45%) of an orange solid (Compound 10).

IR (KBr): 3350, 3200, 1650, 1480, 1450, 1260, 1220, 940, 820, 760, 700cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 2.76 (s, 3H), 4.40 (s, 2H), 5.64 (d, 1H, J = 7.9 Hz), 6.80 (d, 1H, J = 7.5 Hz), 6.87 (d, 1H, J = 7.4 Hz), 7.29-7.39 (m, 3H), 7.46-7.54 (m, 4H), 7.62 (t, 1H J = 7.2 Hz), 7.70 (s, 1H), 7.92 (d , 2H, J = 8.2 Hz), 8.04 (m, 2H), 8.69 (d, 1H, J = 8.2 Hz)

Mass: m / z 463 (M ⁺ ), 335, 266, 223, 196, 169, 147

Elemental Analysis for _{C 28 H 25 N 5 O 2} · H 2 O

Calc. C 69.84; H 5.65; N 14.54

Found C 69.66; H 5.58; N 14.58

Example 4

: N ⁶ -methyl-N ⁶ and N ⁶ -methyl-N ^6- [4- [N- (3-aminocarbonylphenyl) aminosulfonyl] benzyl] -2,6-diaminobenz [cd] indole ( Preparation of Compound 19)

.

1) Preparation of 4- (bromomethyl) benzenesulfonylchloride (Compound 11)

After dissolving 10.0 g (52.5 mmol) of p-toluenesulfonyl chloride in 300 ml of carbon tetrachloride, 10.3 g (57.9 mmol) of NBS (N-bromosuccinimide) and 1.27 g (5.25 mmol) of benzoyl peroxide were added thereto. The reflux was stirred for an hour. After the reaction, the mixture was cooled to room temperature, filtered and the filtrate was concentrated under reduced pressure. The obtained residue was purified by column chromatography using a 10: 1 mixture of hexane and ethyl acetate to give 12.0 g of a colorless liquid. 200 ml of hexane was added to the liquid, and the resulting solid was stirred at -30 ° C. The resulting solid was filtered to yield 9.2 g (yield 65%) of white solid (Compound 11).

Melting Point: 73.8 ~ 74.2 ℃

IR (KBr): 3100, 1950, 1600, 1420, 1380, 1180, 1080, 850, 670 cm ^-1

¹ H NMR (CDCl ₃ ): δ 4.52 (s, 2H), 7.66 (d, 2H, J = 8.5Hz), 8.03 (d, 2H, J = 8.5Hz)

2) Preparation of N- (benzyloxycarbonyl) -3-aminobenzoic acid methyl ester (Compound 12)

Dissolve 10.0 g (66.2 mmol) of 3-aminobenzoic acid methyl ester in 70 mL (70.0 mmol) of 1N sodium hydroxide methanol solution, add dropwise 10.0 mL (70.0 mmol) of benzylchloroformate at 5 ° C. or lower, and then at room temperature for 2 hours. Stirred. The reaction mixture was concentrated under reduced pressure, 100 ml of water was added thereto, and the pH was neutralized to 7 with 1N aqueous hydrochloric acid solution. The reaction mixture was extracted three times with 100 ml of ethyl acetate, washed with brine, and dried over magnesium sulfate. The residue obtained by concentrating the organic solution under reduced pressure was purified by flash column chromatography using methylene chloride to give 17.0 g (yield 90%) of a white solid (Compound 12).

Melting Point: 119.8 ~ 120.0 ℃

¹ H NMR (CDCl ₃ ): δ 3.90 (s, 3H), 5.22 (s, 2H), 6.91 (s, 1H), 7.37 to 7.41 (m, 6H), 7.73 (m, 2H), 7.97 (s, 1H)

3) Preparation of N- (benzyloxycarbonyl) -N- (4- (bromomethyl) benzenesulfonyl) -3-aminobenzoic acid methyl ester (Compound 13)

15.0 g (52.5 mmol) of N- (benzyloxycarbonyl) -3-aminobenzoic acid methyl ester was dissolved in 300 mL of tetrahydrofuran, 1.5 g (63.1 mmol) of sodium hydride were added little by little, followed by stirring for 30 minutes. 14.2 g (52.7 mmol) of 4- (bromomethyl) benzenesulfonyl chloride was further added thereto, followed by stirring at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, 400 ml of methylene chloride was added thereto, the mixture was washed three times with 50 ml of water and once with brine, and dried over magnesium sulfate. The residue obtained by concentrating the organic solution under reduced pressure was purified by column chromatography using a 10: 1 mixture of methylene chloride and ethyl acetate to give 16.5 g (yield 60%) of a white solid (Compound 13).

Melting Point: 152.7 ~ 153.5 ℃

IR (KBr): 3050, 2950, 1750, 1730, 1600, 1450, 1370, 1310, 1270, 1090, 850, 760 cm ^-1

¹ H NMR (CDCl ₃ ): δ 3.94 (s, 3H), 4.50 (s, 2H), 5.10 (s, 2H), 7.13 (m, 2H), 7.30 to 7.32 (m, 3H), 7.49 to 7.55 ( m, 4H), 7.92 (m, 3H), 8.15 (s, 1H)

4) Preparation of N ⁶ (Compound 14)

2.96 g (16.1 mmol) of 6-aminobenz [cd] indole-2 (1H) -one and 6.16 mL (35.3 mmol) of diisopropylethylamine were dissolved in 100 mL of dimethylformamide and N- (benzyloxycarbonyl) 10.0 g (19.3 mmol) of -N- (4- (bromomethyl) benzenesulfonyl) -3-aminobenzoic acid methyl ester was added thereto, followed by reaction at room temperature for 6 hours. After the reaction, the mixture was added to 800 ml of dimethylformamide, and a solid was formed and filtered. The solid was dissolved in methylene chloride, washed with water, dried over sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by flash column chromatography using a 2: 1 mixture of methylene chloride and ethyl acetate to give 7.2 g (yield 72%) of an orange solid (Compound 14).

Melting Point: 177.5 ~ 178.2 ℃

¹ H NMR (DMSO-d ₆ ): δ 3.93 (s, 3H), 4.68 (d, 2H, J = 5.6 Hz), 5.14 (s, 2H), 6.22 (d, 1H, J = 7.8 Hz), 6.76 (d, 1H J = 7.6 Hz), 7.17 (m, 2H), 7.32 (m, 4H), 7.74 (m, 4H), 7.88 (t, 1H, J = 8.3 Hz), 7.96 (d, 2H, J = 8.4 Hz), 8.07 (d, 1H, J = 7.0 Hz), 8.14 (d, 1H, J = 7.2 Hz), 8.61 (d, 1H, J = 8.2 Hz), 10.47 (s, 1H)

Preparation of methyl ^{-N 6 15) - 5) N} 6

5.0 g (8.04 mmol) of N ⁶ and 2.1 mL (12.06 mmol) of diisopropylethylamine were dissolved in 30 mL of dimethylformamide, 0.60 mL (9.70 mmol) of iodine methane was added, followed by stirring at room temperature overnight. After the reaction, the mixture was added to 750 ml of water and filtered when an orange solid formed. This solid was dissolved in methylene chloride, dried over sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by flash column chromatography using a 1: 1 mixture of methylene chloride and ethyl acetate to give 3.16 g (yield 62%) of an orange solid (Compound 15).

IR (KBr): 3450, 3200, 1750, 1700, 1450, 1300, 1170, 750 cm ^-1

¹ H NMR (CDCl ₃ + DMSO-d ₆ ): δ 2.85 (s, 3H), 3.94 (s, 3H), 4.45 (s, 2H), 5.10 (s, 2H), 6.87 (d, 1H, J = 7.5 Hz), 6.94 (d, 1H J = 7.6 Hz), 7.14 (dd, 2H, J = 4.0 Hz), 7.28 (m, 3H), 7.50-7.62 (m, 4H), 7.73 (t, 1H, J) = 7.3 Hz), 7.89 (m, 3H), 8.04 (d, 1H, J = 6.9 Hz), 8.14 (d, 1H, J = 7.2 Hz), 8.25 (d, 1H, J = 8.2 Hz)

Mass: m / z 635 (M ⁺ )

Preparation of methyl ^{-N 6 16) - 6) N} 6

3.0 g (4.72 mmol) of N ⁶ -methyl-N ⁶ were mixed with 100 mL of toluene, heated to 120 ° C., and then 2.1 g (5.19 mmol) of Lawson's reagent was added thereto, and the mixture was stirred for 1 hour. The reaction mixture was concentrated under reduced pressure to remove the solvent, and purified by flash column chromatography using a 3: 1 mixture of methylene chloride and ethyl acetate to obtain 1.54 g (yield 51%) of a red solid (Compound 16).

¹ H NMR (CDCl ₃ + DMSO-d ₆ ): δ 2.93 (s, 3H), 3.94 (s, 3H), 4.57 (s, 2H), 5.14 (s, 2H), 6.97 (d, 1H, J = 7.7 Hz), 7.05 (d, 1H J = 7.7 Hz), 7.17 (dd, 2H, J = 4.0 Hz), 7.32 (m, 3H), 7.64 (m, 4H), 7.72 (t, 1H, J = 8.3 Hz), 7.99 (m, 3H), 8.07 (d, 1H, J = 7.0 Hz), 8.14 (d, 1H, J = 7.2 Hz), 8.28 (d, 1H, J = 8.3 Hz)

7) Manufacture of 17)

1.5 g (2.30 mmol) of N ⁶ -methyl-N ⁶ were dissolved in 10 ml of a 1: 1 mixture of ethanol and tetrahydrofuran, and 5.0 ml (5.0 mmol) of 1N sodium hydroxide solution was added dropwise at room temperature, followed by 0.16 ml of iodine methane ( 2.54 mmol) was added and stirred for 30 minutes. 50 ml of water was added thereto, extracted three times with 70 ml of ethyl acetate, washed with brine, and dried over sodium sulfate. The residue obtained by concentrating the organic solution under reduced pressure was purified by flash column chromatography using a 1: 1 mixture of methylene chloride and ethyl acetate to obtain 0.85 g (yield 70%) of a red solid (Compound 17).

IR (KBr): 3450, 3300, 2950, 1730, 1600, 1500, 1460, 1300, 1150, 1020, 940, 760 cm ^-1

¹ H NMR (CDCl ₃ ): δ 2.83 (s, 3H), 2.85 (s, 3H), 3.85 (s, 3H), 4.53 (s, 2H), 6.82 (d, 1H, J = 7.7Hz), 6.93 (s, 1H), 7.36 (t, 1H J = 7.8 Hz), 7.42 to 7.55 (m, 5H), 7.69 (t, 1H, J = 3.5 Hz), 7.78 to 7.83 (m, 3H), 7.87 (d , 1H, J = 7.0 Hz), 8.04 (d, 1H, J = 8.2 Hz)

Mass: m / z 531 (M ⁺ )

8) N ⁶ -methyl-N ^6- [4- [N- (3-methoxycarbonylphenyl) aminosulfonyl] benzyl] -2,6-diaminobenz [cd] indole (Compound 18) and N ⁶ Preparation of -methyl-N ^6- [4- [N- (3-aminocarbonylphenyl) aminosulfonyl] benzyl] -2,6-diaminobenz [cd] indole (Compound 19)

0.25 g was added to 20 ml of methanol solution saturated with ammonia gas and reacted at 145 ° C. for 5 hours in a sealed container. After the reaction, the mixture was cooled to room temperature and the solvent was concentrated under reduced pressure. The obtained residue was purified by column chromatography using a 5: 1 mixture of methylene chloride and methanol to obtain 0.072 g of red orange solid (Compound 18) (yield 31%) and 0.070 g of orange solid (Compound 19) (yield 32). %) Were each obtained.

[Compound 18]

IR (KBr): 3300, 3100, 2950, 2850, 1720, 1680, 1650, 1600, 1490, 1300, 1210, 1130, 1100, 760 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 2.70 (s, 3H), 3.82 (s, 3H), 4.37 (s, 2H), 6.77 (d, 1H, J = 7.6Hz), 6.87 (d, 1H J = 7.5 Hz), 7.38-7.40 (m, 2H), 7.55-7.74 (m, 4H), 7.70-7.72 (m, 3H), 8.06 (d, 2H, J = 7.6 Hz)

Mass: m / z 500 (M ⁺ ), 317, 269, 241, 226, 211

Elemental Analysis for C ₂₇ H ₂₄ N ₄ O ₄ S

Calc. C 64.78; H 4.83; N 11.19; S 6.41; O 12.78

Found C 64.97; H 4.98; N 11.26; S 6.42; O 12.37

[Compound 19]

IR (KBr): 3400, 3250, 1680, 1540, 1480, 1240, 1090, 710 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 2.74 (s, 3H), 4.36 (s, 2H), 6.78 (d, 1H, J = 7.5 Hz), 6.88 (d, 1H J = 7.5 Hz), 7.25- 7.30 (m, 2H), 7.51 (s, 1H), 7.54-7.74 (m, 5H), 7.77 (d, 2H, J = 8.4 Hz), 7.87 (s, 1H), 8.06 (m, 2H)

Mass: m / z 485, 310, 282, 256, 224, 196, 169

Elemental Analysis for C ₂₆ H ₂₃ N ₅ O ₃ S · H ₂ O

Calc. C 62.01; H 5.00; N 13.91; S 6.37; O 12.71

Found C 62.40; H 4.94; N 13.62; S 6.57; O 12.47

Example 5

: N ^6-methyl -N ⁶ - [4 - [(phenyl-sulfonyl-aminocarbonyl-benzyl)] - Preparation of 2,6-diamino-benz [cd] indole (Compound 21)

.

1) Manufacture of 20)

1.0 g (2.76 mmol) of N-methyl-N- (4-hydroxycarbonylbenzyl) -2-methylthio-6-aminobenz [cd] indole was dissolved in 50 ml of tetrahydrofuran and 1,1′-carbox 0.45 g (2.76 mmol) of carbonyldiimidazole was added and stirred at 50 ° C. for 3 hours. Tetrahydro dissolved 0.43 g (2.76 mmol) of benzenesulfonamide (Aldrich 10,814-6) and 0.41 mL (2.76 mmol) of DBU (1,8-diazabicyclo [5,4,0] -7-undecene) 10 ml of furan solution was added, followed by stirring at 50 ° C. for 12 hours. After the reaction, the mixture was concentrated under reduced pressure, and the residue was purified by column chromatography using a 5: 1 mixture of tetrahydrofuran and methanol to obtain 0.55 g (yield 40%) of a red solid (Compound 20).

IR (KBr): 3300, 3100, 2950, 2850, 1680, 1600, 1550, 1460, 1350, 1250, 1140, 1090 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 2.77 (s, 3H), 2.89 (s, 3H), 4.61 (s, 2H), 6.84 (d, 1H, J = 7.7Hz), 7.19 (s, 1H) , 7.35 (d, 2H J = 8.2 Hz), 7.43 (m, 3H), 7.58 (d, 1H, J = 7.6 Hz), 7.61 (t, 1H, J = 7.2 Hz), 7.84 (m, 2H), 7.87 (d, 2H, J = 8.2 Hz), 7.96 (d, 1H, J = 7.1 Hz), 8.18 (d, 1H, J = 8.1 Hz)

Mass: m / z 502, 485, 466, 430, 383, 325, 306

2) N ⁶ - Preparation of [4- (phenylsulfonyl-amino-carbonyl) benzyl] -2,6-diamino-benz [cd] indole (compound 21) ^-N-methyl-6

0.25 g of N-methyl-N- [4- (phenylsulfonylaminocarbonyl) benzyl] -2-methylthio-6-aminobenz [cd] indole is added to 25 ml of methanol solution saturated with ammonia gas and The reaction was carried out at 145 ° C. for 5 hours. After the reaction, the mixture was cooled to room temperature and the solvent was concentrated under reduced pressure. The obtained residue was purified by column chromatography using a 3: 1 mixture of methylene chloride and methanol to give 0.10 g (yield 51%) of a red solid (Compound 21).

Melting Point: 180 ℃ (Decomposition)

IR (KBr): 3350, 3240, 1640, 1600, 1540, 1350, 1130, 1080, 820, 600cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 2.73 (s, 3H), 4.35 (s, 3H), 6.80 (d, 1H, J = 7.6Hz), 6.93 (d, 1H J = 7.5Hz), 7.33 ( d, 2H, J = 8.1 Hz), 7.39 (m, 3H), 7.62 (t, 1H, J = 6.6 Hz), 7.83 (m, 2H), 7.89 (d, 2H, J = 8.1 Hz), 8.10 ( d, 1H, J = 7.0 Hz, 8.15 (d, 1H, J = 8.2 Hz)

Mass: m / z 471 (M ⁺ )

Elemental Analysis for C ₂₆ H ₂₂ N ₄ O ₃ S

Calc. C 66.37; H 4.71; N 11.91; S 6.81

Found C 66.42; H 4.80; N 11.73; S 6.69

Experimental Example: In vitro cancer cell proliferation inhibitory effect

The anticancer activity of the compounds according to the present invention was verified by measuring the effect of cell proliferation inhibition in vitro by the following method. This study included mouse-derived lymphoid leukemia cells L1210 (ATCC CCL-219), human-derived T-cell acute lymphocytic leukemia cells CCRF-CEM (ATCC CRL-8436), colon adenocarcinoma cells SW480 (ATCC CCL-228), and Cancer cells of a species of breast adenocarcinoma cell MCF7 (ATCC HTB-22) were used.

As a medium for cell culture, RPMI-1640 medium containing 5% calf serum and 0.5% DMSO (dimethylsulfoxide) was used and cultured in a constant temperature and humidity 5% CO ₂ incubator at 37 ° C. Cell passage was performed once every 3 to 4 days.

For the measurement of anticancer activity, MTT (= 3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetra) was used to determine the concentration at which cell growth was inhibited to 50% of the untreated group (ED ₅₀ ). Zolium bromide] spectroscopy, cells were first divided into 6,000 (L1210), 10,000 (CCRF-CEM), 8,000 (SW480) and 6,000 (MCF7) cells in each well of a 96-well plate. Cell growth was monitored by varying the concentration of the test compound, DMSO was used as needed to dissolve the compounds, and the concentration of DMSO added to the cell culture solution was adjusted to 0.5% or less. L1210), 3 days (CCRF-CEM, SW480), 4 days (MCF7), treated with MTT for 4 hours after incubation, and cells were collected. The resulting precipitated formazan was dissolved in DMSO and then microplate reader (m) at 540 nm. Absorbance was measured using an icroplate reader.

To obtain the ED ₅₀ value, the concentration causing 50% inhibition was calculated from a graph of compound concentration logarithm versus cell growth percentage. These ED ₅₀ values were taken three times and averaged. The anticancer activity of the test compound is shown in Table 1 below.

From the experimental results described in Table 1, it can be seen that the compound according to the present invention can be effectively used as an anticancer agent by effectively inhibiting the proliferation of cancer cells.

Claims (6)

Benzindole derivatives of the general formula (I) and pharmaceutically acceptable salts thereof

Wherein R represents a group of -CO-R ₁ , -CONH-R ₂ or -SO ₂ -NH-R ₂ , wherein R ₁ is unsubstituted or substituted by alkyl and in nitrogen, sulfur and oxygen A mono- or di-heterocyclic group containing at least one hetero atom selected, R ₂ is aryl unsubstituted or substituted by hydroxycarbonyl, aminocarbonyl or alkoxycarbonyl,

R ₃ represents aryl and R ₄ represents carboxyl, aminocarbonyl or alkoxycarbonyl, or arylsulfonyl. The compound of claim 1, wherein R is hydroxycarbonylphenylaminocarbonyl, aminocarbonylphenylaminocarbonyl, methoxycarbonylphenylaminocarbonyl, α-methoxycarbonylbenzylaminocarbonyl, α-hydroxycarbon Carbonylbenzylaminocarbonyl, α-aminocarbonylbenzylaminocarbonyl, hydroxycarbonylphenylaminosulfonyl, aminocarbonylphenylaminosulfonyl, methoxycarbonylphenylaminosulfonyl, α-hydroxycarbonylbenzylamino A compound of formula (I) which is sulfonyl, α-aminocarbonylbenzylaminosulfonyl, α-methoxycarbonylbenzylaminosulfonyl, phenylsulfonylaminocarbonyl or phenylcarbonylaminosulfonyl and pharmaceutically Acceptable salts. The compound of claim 2, wherein R is aminocarbonylphenylaminocarbonyl, methoxycarbonylphenylaminocarbonyl, α-aminocarbonylbenzylaminocarbonyl, α-methoxycarbonylbenzylaminocarbonyl, aminocarbonylphenyl A compound of formula (I) which is aminosulfonyl or methoxycarbonylphenylaminosulfonyl and a pharmaceutically acceptable salt thereof. The following compound of general formula (II) is alkylated with the compound of general formula (III), and then alkylated with the compound of general formula (IV) to prepare a compound of general formula (V) using Lawson reagent. And reacting the compound of formula (V) with the compound of formula (IV) to produce methylthio compound (VI), followed by ammonia substitution reaction, wherein the benzindole derivative of formula (I) and Method for preparing its salts.

.

Wherein R, R ₁ and R ₂ are as defined in claim 1 wherein R 'has the same meaning as R or represents an alkyl-substituted ester or sulfonylamide, provided that If present, a protecting group can be introduced into the amide nitrogen, and X represents a reactive leaving group. The method according to claim 4, wherein the protecting group introduced into the amide nitrogen of R 'is benzyl, benzyloxy, carbenzyloxy, benzyloxymethyl, t-butoxycarbonyl or t-butyldimethylsilyloxymethyl. An anticancer composition comprising the compound of formula (I) according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof as an active ingredient.