KR100563195B1 - Novel benzofluorene dione derivatives - Google Patents

Abstract

The present invention relates to a compound represented by the following formula (1), a pharmaceutically acceptable salt thereof, a preparation method thereof and a pharmaceutical composition thereof. Since the compound effectively inhibits cell proliferation, the compound may be used as a prophylactic, therapeutic and therapeutic aid for various cancers.

[Formula 1]

In the above formula, the definition of ring A and ring B is as defined in the specification of the present invention.

Benzofluorene, anticancer, cytotoxicity

Description

Novel benzofluorene dione compounds {NOVEL BENZOFLUORENE DIONE DERIVATIVES}

Quinone compounds generally have various physiological activities [RA Morton, Ed. Biochemistry of Quinones; Academic Press: New York, 1965], among which heterocyclic quinones are known to have excellent anticancer activity [GA Efimove, LS Efres, Zh . Org . Khim . , 1967, 3 , 162].

Suh et al. Reacted 6,7-dichloroquinoline-5,8-dione with 2-aminopyridine derivatives for the purpose of developing a new quinone anticancer agent. ] Fluorene-5,6-dione derivatives were synthesized and the cytotoxic effects of these compounds were measured on five human tumor cell lines. As a result, 2-methyl-4a, 10,11-triaza-benzo [3,2-a] fluorene-5,6-dione is an anticancer agent currently used against human ovarian cancer cell line (SK-OV-3). It was found that the anticancer effect is superior to that of phosphorus doxorubicin [Y. S, Kim, SY Park, ME Suh, HJ Lee, D. Schollmeyer, Bio. Org . Chem ., 2003, 11 , 1829.

On the other hand, ortho-quinoline derivatives such as triaza-benzo [3,2-a] fluorene-5,6-dione derivatives inhibit the action of avian myeloblastosis virus reverse transcriptase (AMV-RT). The degree of inhibition is generally reported to be much better than para-quinoline derivatives [Y. Inouye, H. Matsumoto, R, Morishige, Y. Kitahara, A. Kabo, S. Nakamira, Chem . Pharm . Bull. , 1991, 39 , 994].

The present inventors have completed the present invention by finding a novel benzofluorene dione compound having excellent anticancer function by showing excellent cytotoxic effect from the above results.

It is an object of the present invention to provide a novel benzofluorene dione compound, a pharmaceutically acceptable salt thereof, a method for preparing the compound and an anticancer pharmaceutical composition containing the compound or its salt as an active ingredient.

The present invention provides a compound represented by the formula (1) and a pharmaceutically acceptable salt thereof.

,

,

,

, 및

등으로 이루어지는 군으로부터 선택되는 1종일 수 있고, B환은

,

및

등으로 이루어지는 군으로부터 선택되는 1종일 수 있다.In the formula, ring A is

,

,

,

, And

It may be one selected from the group consisting of and the like, ring B is

,

And

It may be one selected from the group consisting of.

In the display of the A ring and the B ring, the side indicated by the broken line represents the side fused to the quinone ring and the imidazole ring in the structure of Chemical Formula 1, respectively. In addition, R used to define the B ring may be one selected from the group consisting of hydrogen, a lower alkyl group, a halogen atom, and the like, and R ₁ may be hydrogen or a lower alkyl group.

In the definitions of R and R ₁ used to denote the B ring throughout the specification including Formula 1 of the present invention, the term "lower alkyl group" refers to a straight or branched chain alkyl group consisting of 1 to 6 carbon atoms. Examples include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, secondary butyl, pentyl, isopentyl, neopentyl, tertiary pentyl, 1-methylbutyl, 2-methylbutyl, 1,2-dimethylpropyl , Hexyl, isohexyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2 , 3-dimethylbutyl, 3,3-dimethylbutyl, 1-ethylbutyl, 2-ethylbutyl, 1,1,2-trimethylpropyl, 1,2,2-trimethylpropyl, 1-ethyl-1-metalpropyl and There is a 1-ethyl-2-methylpropyl group, and the term “halogen atom” used in the definition of R means fluorine, chlorine, bromine and iodine.

또는

이고, 상기 B환의 정의에 사용된 상기 R은 수소, 메틸기, 에틸기, 프로필기, 이소프로필기, 불소, 염소 및 브롬으로 이루어지는 군으로부터 선택되는 1종이며, 상기 R₁은 수소, 메틸기, 에틸기, 프로필기 및 이소프로필기로 이루어지는 군으로부터 선택되는 1종인 화합물들이다.Preferred examples of the compound represented by Formula 1 is the A ring of Formula 1

or

R is used in the definition of the ring B is one selected from the group consisting of hydrogen, methyl, ethyl, propyl, isopropyl, fluorine, chlorine and bromine, wherein R ₁ is hydrogen, methyl, ethyl, It is 1 type of compounds chosen from the group which consists of a propyl group and an isopropyl group.

In addition, specific examples of the compound represented by Formula 1 include the following compounds.

Viii) 10-methyl-1,6b, 11-triaza-benzo [3,2-a] fluorene-5,6-dione

Ii) 7-methyl-1,6b, 11-triaza-benzo [3,2-a] fluorene-5,6-dione

Viii) 8-chloro-1,6b, 11-triaza-benzo [3,2-a] fluorene-5,6-dione

Viii) 1,6b, 10,11-tetraaza-benzo [a] fluorene-5,6-dione

Viii) 1,6b, 9,11-tetraaza-benzo [a] fluorene-5,6-dione

Viii) 2,3,6b, 11-tetraaza-benzo [a] fluorene-5,6-dione

Viii) 8-methyl-2,3,6b, 11-tetraaza-benzo [a] fluorene-5,6-dione

Viii) 10-methyl-2,3,6b, 11-tetraaza-benzo [a] fluorene-5,6-dione

The present invention also provides a method for preparing the compound represented by the formula (1).

1 is a schematic view showing a method for preparing a compound of the present invention, and the following Chemical Formulas 2 and 3 are starting materials for preparing the compound of the present invention.

1

In the figure, the definition of ring A and ring B is as in Chemical Formulas 1, 2 and 3.

,

,

,

, 및

등으로 이루어지는 군으로부터 선택되는 1종이다. 또한, 상기 A환을 표시하는데 있어서, 파선으로 표시된 변은 상기 화학식 2의 구조 중 퀴논고리와 융합되어(fused) 결합된 변을 표시한 것이다.In the formula, ring A is

,

,

,

, And

It is 1 type chosen from the group which consists of etc. In addition, in displaying the A ring, the side indicated by the broken line represents the side fused with the quinone ring in the structure of Chemical Formula 2.

,

및

등으로 이루어지는 군으로부터 선택되는 1종이고, 상기 B환을 정의하는데 사용된 R은 수소, 저급알킬기 및 할로겐원자로 이루어지는 군으로부터 선택되는 1종이며, R₁은 수소 또는 저급알킬기이다. 또한, 상기 B환을 표시하는데 있어서, 파선으로 표시된 변은 상기 화학식 3의 구조 중 C=N 결합이 이루는 한 변을 표시한 것이다.In the above formula, ring B is

,

And

It is 1 type chosen from the group which consists of etc., R used for defining the said B ring is 1 type chosen from the group which consists of hydrogen, a lower alkyl group, and a halogen atom, and R <_1> is hydrogen or a lower alkyl group. In addition, in the B ring, the side indicated by the broken line represents one side of the C = N bond in the structure of Chemical Formula 3.

Hereinafter, the manufacturing method of the present invention will be described in more detail.

Starting compound of formula (2) and catalyst anhydrous potassium carbonate (K ₂ CO ₃ ) is dissolved in a suitable alcohol solvent and then reacted by adding the compound of formula ( ₃ ) to the solution. Preferred examples of the alcohol solvent used in the reaction include methyl alcohol, ethyl alcohol, propyl alcohol, butyl alcohol, and the like, in particular ethyl alcohol or isopropyl alcohol. In this case, the molar ratio of the compound of Formula 2 and the compound of Formula 3 may be such that the equivalent ratio to the compound of Formula 3 is an excess equivalent, particularly, 1: 1 to 1: 2.5 are preferred, and 1: 1.1 to 1: 2.0 are most preferred. Do. The anhydrous potassium carbonate used as a catalyst may be used so as to 0.7 to 1 equivalent to the compound of formula (2). In the reaction, the reaction temperature is preferably a reflux temperature and the reaction time may be given a sufficient time for the reaction to occur for each compound, preferably 10 hours to 70 hours or more preferably 15 hours to 55 hours Can react.

After the reaction may be subjected to a post-treatment process, such as appropriate extraction or purification known to those skilled in the art or known to those skilled in the art as needed. Preferred examples of the post-treatment will be shown appropriately in the examples described below.

Pharmaceutically acceptable salts of the compounds of the invention are, for example, acid addition salts of the compounds of the invention which are sufficiently basic, for example inorganic or organic acids (eg hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid). , Benzenesulfonic acid, trifluoroacetic acid, citric acid, lactic acid or maleic acid). In addition, pharmaceutically acceptable base addition salts of the compounds of the present invention that are sufficiently acidic include alkali metal salts such as lithium, sodium or potassium salts; Alkaline earth metal salts such as calcium or magnesium salts; Ammonium salts; Or salts with organic bases that provide physiologically acceptable cations, for example with methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris- (2-hydroxyethyl) amine. All the salts fall within the scope of the present invention and they can be prepared by conventional methods. For example, they can be prepared by contacting acidic and basic groups in a suitable aqueous, non-aqueous or partially aqueous medium, generally in a stoichiometric ratio. The salts can be recovered by filtration as appropriate, filtration after precipitation with non-solvents, preferably ethereal or hydrocarbon solvents, evaporation of the solvent, or lyophilization in the case of aqueous solvents.

The present invention also provides a pharmaceutical composition for anticancer containing the compound represented by the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

The anticancer pharmaceutical composition of the present invention can be used in mammals, especially humans, for the purpose of preventing, treating or assisting in hyperproliferation (for example, cancer diseases). In particular, various benign or malignant human tumors (lung cancer, ovarian cancer, skin cancer, melanoma cancer, central nerve cancer, colon cancer, kidney cancer, liver cancer, bladder cancer, breast cancer, stomach cancer, uterine cancer, prostate cancer, colon cancer, vulvar cancer, thyroid cancer, sarcoma) , Glioblastoma, various head and neck tumors, etc.), and particularly preferably lung, ovarian cancer, melanoma cancer, central nervous system cancer, or colon cancer, or as a therapeutic or adjuvant, prophylactic for the treatment of colon cancer.

The pharmaceutical compositions of the present invention may be prepared in unit dosage form by formulation with a pharmaceutically acceptable carrier or excipient or may be prepared in a multi-dose container and may be oral, parenteral or single or multiple times. It may be administered topically.

As used herein, the term “parenteral” includes, without limitation, subcutaneous, intravenous, arterial or infusion techniques. The term "topical" also includes inhalation spray and rectal administration, as well as administration by conventional routes such as mucous membranes and skin of the mouth and nose.

The pharmaceutical composition may be in solid, semisolid or liquid dosage form (e.g. in the form of tablets, capsules, pills, powders, suppositories, solutions, elixirs, syrups, suspensions, creams, sugar-containing tablets, pastes and sprays) Can be. As is known in the art, the route of administration of the phthalazine derivatives described herein is selected according to the type of composition to be used. In general, it is preferred to use unit dosage forms of the inhibitors described herein to facilitate the treatment and administration of the correct dose of the active compound. In general, the compounds having a therapeutic effect of the present invention are present in a dosage form that is sufficient to provide a desired level of concentration, i.

The pharmaceutical composition of the present invention is generally administered in a single or multidose form in an amount of about 0.01 to 100 mg / kg per day based on the active ingredient, but the dosage may be determined differently according to the health condition or pathological findings of the individual have.

For oral administration, tablets containing various excipients such as sodium citrate, calcium carbonate and dicalcium phosphate may contain various disintegrants such as starch, preferably potato or tapioca starch, alginic acid and certain silicate complexes, polyvinyl pyrrolidone, It can be used with binders such as sucrose, gelatin and gum arabic. In addition, lubricants such as magnesium stearate, sodium lauryl sulfate and talc are sometimes very useful for tableting. Solid compositions of a similar type may also be used as fillers in soft and hard-filled gelatin capsules. Preferred materials related to this include high molecular weight polyethylene glycols as well as lactose or lactose. If oral administration of an aqueous suspending agent or elixir is desired, the essential active ingredients contained therein are various sweetening or flavoring agents, colorants or pigments and, if desired, emulsifying and / or suspending agents as well as water, ethanol, propylene glycol, It can be used in combination with diluents such as glycerin and various similar mixtures.

For parenteral administration, various substances can be used as carriers, adjuvants and vehicles in the compositions of the present invention as useful in the pharmaceutical sector. Injections, such as oily solutions, suspensions or emulsions, may be formulated by known techniques using the necessary suitable dispersing or wetting agents and suspending agents. Sterile injectables may also be used, for example, sterile nonpyrogenic or parenterally acceptable non-toxic diluents or solvents such as 1,3-butanediol. Other acceptable vehicles and solvents that may be used include 5% Textose Injection, Ringer's Injection and Sodium Chloride Injection. Sterile, nonvolatile oils are also commonly used as solvents or suspending media. Any mixed nonvolatile oil may be used for this purpose, including synthetic mono-, di- or triglycerides. Fatty acids such as oleic acid may also be used in the preparation of injectable compositions.

Suppositories for rectal administration of the pharmaceutical composition of the present invention may be prepared by mixing the drug with a suitable non-irritating excipient such as cocoa butter, polyethylene glycol, etc., which is solid at room temperature but liquid at body temperature and thus melts in the rectum to release the drug.

In addition, the pharmaceutical composition of the present invention may be administered locally, and for this purpose, it is preferable to administer it in cream, salves, jelly, paste, ointment, and the like according to standard pharmaceutical practice.

The following examples are intended to illustrate the invention by illustrating the starting materials for carrying out the invention and the preparation of compounds of the invention and measuring their cytotoxic effects, which are not intended to limit the scope of the invention. .

EXAMPLE

Example  1) 10- methyl -1,6b, 11- Triaza - Benzo [3,2-a] fluorene -5,6- Dion

1-a) 6,7- Dichloroquinoline -5,8- Dion's  Produce

6,7-dichloroquinoline-5,8-dione was prepared by the method published in the paper [CW Schellhammer, S. Peteren, G. Domagk, Ann , 1959, 624 , 108].

1-a-1) 5-nitro-8-hydroxyquinolineHCl

29 g (0.2 ml) of 8-hydroxyquinoline (Merch, Dalstadt, Germany) was added to 100 ml of water and 100 g of ice and stirred, and 37.5 ml of concentrated hydrochloric acid (37%) was added thereto and stirred for 1 hour in an ice bath. It was. An aqueous solution of 15 g of NaNO ₂ (sodium nitrite) dissolved in 25 mL of water was slowly added dropwise to the mixed solution obtained by using a dropping funnel while maintaining 0-4 ° C. The mixture obtained in the above process was left at 0 ° C. for 24 hours, then maintained, and the resulting precipitate was suction filtered and washed several times with ice water to obtain a pH 5-6 to obtain yellow solid crystals, which were dried at room temperature.

Melting Point: 250 ℃ or higher

1-a-2) 5-amino-8-hydroxyquinolineH ₂ SO ₄

40 g (0.2 mol) of 5-nitro-8-hydroxyquinoline-HCl obtained in the above process was dissolved in 160 ml of water, and 260 ml of 5N-NaOH solution was added thereto, followed by heating to 55 ° C and stirring. 95 g of a reducing agent Na ₂ S ₂ O ₄ (sodium hydrosulfite) were added to the reaction mixture at a time so that the temperature of the reaction spontaneously reached 90-95 ° C. and maintained at 75 ° C. for 1 hour. After passing the N ₂ gas through the reactant and bubbling to cool to 50 ° C., 250 mL of 12N-H ₂ SO ₄ was slowly added thereto, and the resulting SO ₂ and SO ₃ gas were removed under reduced pressure. The resulting precipitate was suction filtered to give red yellow crystals which were dried at room temperature.

 Melting Point: 250 ℃ or higher

1-a-3) 6,7-dichloroquinoline-5,8-dione

To 40 g (0.15 mol) of 5-amino-8-hydroxyquinoline-H ₂ SO ₄ obtained in the above process, 50 ml of concentrated hydrochloric acid was added, and the mixture was heated and stirred to 55 ° C. To the reaction mixture obtained in the above process, a solution of 20 g of NaClO ₃ (sodium chlorate) dissolved in 30 ml of a minimum amount of water was slowly added dropwise. After stirring the mixture of the above process for 30 minutes while maintaining 50-60 ℃, the mixture was poured into ice water and the resulting precipitate was filtered by suction and reduced pressure and washed with ice water. The product obtained at this time was recrystallized with ethanol to give a pale yellow powder, which was dried at room temperature.

Yield: 30g (90% or more), Melting Point: 220-222 ℃

1-b) 10- methyl -1,6b, 11- Triaza - Benzo [3,2-a] fluorene -5,6- Dion

1.0 g (4.7 mmol) of 6,7-dichloroquinoline-5,8-dione and 0.6 g (4.4 mmol) of anhydrous K ₂ CO ₃ were added to 50 mL of anhydrous ethanol and dissolved. 0.7 mL (6.2 mmol) of 2-amino-4-methyl-pyridine (Aldrich, Wisconsin, USA) was added to the mixture, followed by heating to reflux for 50 hours. The reaction solution was cooled and the precipitated crystals were filtered under reduced pressure and recrystallized with ethanol to give an orange solid.

Yield: 0.352g (28%) Melting Point: 300 ℃ or higher

IR (KBr, cm ^-1 ): 1650 (C = O)

¹ H-NMR (CDCl ₃ ): 9.17 (br. D, J 6.8 Hz, 1H), 8.90 (dd, J 4.8 and 1.2 Hz, 1H), 8.35 (dd, J 7.6 and 1.2 Hz, 1H), 7.44 ( m, 2H), 7.12 (t, J 6.8 Hz, 1H), 2.75 (s, 3H)

Elemental analysis: C ₁₅ H ₉ N ₃ O ₂ (MW: 263.25)

Calculated Value: C 68.44 H 3.45 N 15.96

Measured value: C 68.58 H 3.45 N 15.61

Example  2) 7- methyl -1,6b, 11- Triaza - Benzo [3,2-a] fluorene -5,6- Dion

1.0 g (4.7 mmol) of 6,7-dichloroquinoline-5,8-dione and 0.6 g (4.4 mmol) of anhydrous K ₂ CO ₃ were added to 50 mL of anhydrous ethanol and dissolved. 0.7 g (6.3 mmol) of 2-amino-6-methyl-pyridine (Aldrich, Wisconsin, USA) was added to the mixture, followed by heating to reflux for 50 hours. After cooling the reaction solution, the precipitated crystals were filtered under reduced pressure and recrystallized with ethanol to give a dark yellow solid.

Yield: 0.623g (50%) Melting Point: 300 ℃ or higher

IR (KBr, cm ^-1 ): 1649 (C = O)

¹ H-NMR (CDCl ₃ ): 9.01 (dd, J 4.8 and 1.6 Hz, 1H), 8.55 (dd, J 8.0 and 1.6 Hz, 1H), 7.77 (d, J 8.8 Hz, 1H), 7.64 (dd, J 8.0 and 4.8 Hz, 1H), 7.54 (dd, J 8.8 and 7.2 Hz, 1H 1H), 6.99 (d, J 7.2 Hz, 1H), 3.11 (s, 3H)

Elemental analysis: C ₁₅ H ₉ N ₃ O ₂ (MW: 263.25)

Calculated Value: C 68.44 H 3.45 N 15.96

Measured value: C 67.97 H 3.47 N 15.69

Example  3) 8- Chloro -1,6b, 11- Triaza - Benzo [3,2-a] fluorene -5,6- Dion

1.0 g (4.7 mmol) of 6,7-dichloroquinoline-5,8-dione and 0.6 g (4.4 mmol) of anhydrous K ₂ CO ₃ were added to 50 mL of anhydrous isopropyl alcohol and dissolved. 1.2 g (9.2 mmol) of 2-amino-5-chloropyridine (Aldrich, Wisconsin, USA) was added to the mixture, followed by heating to reflux for 34 hours. After cooling the reaction solution and removing the solvent, the resulting material was extracted with methylene chloride (CH ₂ Cl ₂ ), dried over anhydrous MgSO ₄ and concentrated, and the obtained material was recrystallized with ethanol to give an orange solid.

Yield: 0.596 g (45%) Melting Point: 300 ℃ or higher

IR (KBr, cm ^-1 ): 1649 (C = O)

¹ H-NMR (CDCl ₃ ): 9.32 (dd, J 2.0 and 0.6 Hz, 1H), 8.88 (dd, J 4.8 and 1.6 Hz, 1H), 8.36 (dd, J 7.6 and 1.6 Hz, 1H), 7.84 ( dd, J 9.2 and 0.6 Hz, 1H), 7.61 (dd, J 9.2 and 2.0 Hz, 1H), 7.46 (dd, J 7.6 and 4.8 Hz, 1H).

Elemental analysis: C ₁₄ H ₆ ClN ₃ O ₂ (MW: 283.67)

Calculated Value: C 59.28 H 2.13 N 14.81

Measured value: C 59.00 H 2.11 N 14.56

Example  4) 1,6b, 10,11- Tetraaza - Benzo [a] fluorene -5,6- Dion

1.0 g (4.7 mmol) of 6,7-dichloroquinoline-5,8-dione and 0.6 g (4.4 mmol) of anhydrous K ₂ CO ₃ were added to 50 mL of anhydrous isopropyl alcohol and dissolved. 0.6 g (6.3 mmol) of 2-amino-pyrimidine (Aldrich, Wisconsin, USA) was added to the mixture, followed by heating to reflux for 48 hours. After cooling the reaction solution and removing the solvent, the resulting material was extracted with methylene chloride (CH ₂ Cl ₂ ), dried over anhydrous MgSO ₄ and concentrated to obtain a column chromatography (ethyl acetate: n-hexane: methanol =). 4: 4: 1), and then recrystallized with ethanol to give a yellow solid.

Yield: 0.145g (12%) Melting Point: 300 ℃ or higher

* IR (KBr, cm ^-1 ): 1649 (C = O)

¹ H-NMR (CDCl ₃ ): 9.49 (dd, J 6.4 and 1.2 Hz, 1H), 8.94 (dd, J 4.8 and 1.2 Hz, 1H), 8.89 (dd, J 8.0 and 1.2 Hz, 1H), 8.38 ( dd, J 8.0 and 4.8 Hz, 1H), 7.49 (dd, J 6.4 and 4.0 Hz, 1H), 7.28 (dd, J 6.4 and 4.0 Hz, 1H).

Elemental analysis: C ₁₃ H ₆ N ₄ O ₂ (MW: 250.21)

Calculated Value: C 62.40 H 2.42 N 22.39

Measured value: C 62.25 H 2.36 N 22.14

Example  5) 1,6b, 9,11- Tetraaza - Benzo [a] fluorene -5,6- Dion

1.0 g (4.7 mmol) of 6,7-dichloroquinoline-5,8-dione and 0.6 g (4.4 mmol) of anhydrous K ₂ CO ₃ were added to 50 mL of anhydrous isopropyl alcohol and dissolved. 0.6 g (6.2 mmol) of 2-amino-pyrazine (Aldrich, Wisconsin, USA) was added to the mixture, followed by heating to reflux for 50 hours. After cooling the reaction solution and removing the solvent, the resulting material was extracted with methylene chloride (CH ₂ Cl ₂ ), dried over anhydrous MgSO ₄ and concentrated to obtain a column chromatography (ethyl acetate: n-hexane: methanol =). 3: 3: 1), and then recrystallized with methanol to give a yellow solid.

Yield: 0.102g (8%) Melting Point: 300 ℃ or more

IR (KBr, cm ^-1 ): 1649 (C = O)

¹ H-NMR (CDCl ₃ ): 9.43 (d, J 1.6 Hz, 1H), 9.13 (dd, J 4.4 and 1.6 Hz, 1H), 8.98 (dd, J 4.8 and 1.6 Hz, 1H), 8.44 (dd, J 8.0 and 1.6 Hz, 1H), 8.40 (d, J 4.4 Hz, 1H), 7.55 (dd, J 8.0 and 1.6 Hz, 1H).

Elemental analysis: C ₁₃ H ₆ N ₄ O ₂ (MW: 250.21)

Calculated Value: C 62.40 H 2.42 N 22.39

Measured value: C 62.87 H 2.55 N 22.34

Example  6) 2,3,6b, 11- Tetraaza - Benzo [a] fluorene -5,6- Dion

6-a) 6,7- Dichlorophthalazine -5,8- Dion's  Produce

6,7-dichlorophthalazine-5,8-dione was prepared by the method published in the paper [JS Kim, KJ Shin, DC Kim, YK Kang, DJ Kim, KH Yoo and SW Park, Bull. Korean Chem . Soc . 2002, 23 , 1425].

[Degree ]

This figure is a schematic diagram showing a method for preparing 6,7-dichloro-5,8-phthalazinedione. Hereinafter, the manufacturing process will be described in more detail.

6-a-1) 5-nitrophthalazine

Dissolve phthalazine (Merck, Darmstadt, Germany) (20.0 g, 0.15 mol) in concentrated sulfuric acid (150 ml) and add potassium nitrate (73.2 g, 0.72 mol) to the solution at 0 ° C in small portions. And it was heated again for 2 days while maintaining 55-60 ℃. The mixture was diluted with water at 0 ° C., then neutralized with sodium hydroxide solution and the precipitate was filtered to give 5-nitrophthalazine.

6-a-2) 5-nitro-8-aminophthalazine

To the 5-nitrophthalazine (26.0 g, 0.15 mol) ethanol (1.5 L) solution HONH _{2 ·} HCl (62.0 g, 0.89 mol) methanol (1 L) solution was gradually added at 50-55 ℃ for 3 hours And stirred. The reaction mixture was cooled with ice water and the resulting precipitate was filtered to give 5-nitro-8-aminophthalazine.

6-a-3) 5,8-diaminophthalazine

5-Nitro-8-aminophthalazine (14.2 g, 74.7 mmol) obtained above was dissolved in methanol (700 ml), and then FeCl ₃ (70.0 mg, 0.43 mmol) and charcoal (4.50) were added to the solution. g) joined. The mixture was refluxed for 5 hours and filtered using celite to give 5,8-diaminophthalazine.

6-a-4) 6,7-dichloro-5,8-phthalazinedione

The 5,8-diaminophthalazine (7.09g, 44.3mmol) obtained above was dissolved in concentrated hydrochloric acid (90.0ml), and then several times each part of NaClO ₃ (6.08g, 49.6mmol) was added to the solution at 0 ° C. Subscribed across. The reaction mixture was stirred for 1 hour at room temperature, cooled by addition of water and the precipitate was filtered to give 6,7-dichloro-5,8-phthalazinedione.

6-b) 2,3,6b, 11- Tetraaza - Benzo [a] fluorene -5,6- Dion

0.5 g (2.2 mmol) of 6,7-dichlorophthalazine-5,8-dione and 0.3 g (2.2 mmol) of anhydrous K ₂ CO ₃ were added to 30 mL of anhydrous ethanol and dissolved. 0.4 g (4.3 mmol) of 2-aminopyridine (Aldrich, Wisconsin, USA) was added to the mixture, followed by heating to reflux for 18 hours. The reaction solution was cooled, the solvent was removed, the resulting material was extracted with methylene chloride (CH ₂ Cl ₂ ), dried over anhydrous MgSO ₄ , and concentrated. The obtained material was purified by column chromatography (ethyl acetate: n-hexane: methanol). = 4: 4: 3) and then recrystallized with ethanol to give a maroon solid.

Yield: 0.112g (20%) Melting Point: 300 ℃ or more

IR (KBr, cm ^-1 ): 1649 (C = O)

¹ H-NMR (CDCl ₃ ): 10.01 (s, 1H), 9.64 (s, 1H), 9.34 (d, J 6.8 Hz, 1H), 7.66 (d, J 8.8 Hz, 1H), 7.78 (t, J 8.8 Hz, 1H), 7.36 (t, J 6.8 Hz, 1H)

Elemental analysis: C ₁₃ H ₆ N ₄ O ₂ (MW: 250.21)

Calculated Value: C 62.40 H 2.42 N 22.39

Measured value: C 62.87 H 2.55 N 22.34

Example  7) 8- methyl -2,3,6b, 11- Tetraaza - Benzo [a] fluorene -5,6- Dion

0.5 g (2.2 mmol) of 6,7-dichlorophthalazine-5,8-dione and 0.3 g (2.2 mmol) of anhydrous K ₂ CO ₃ were added to 30 mL of anhydrous ethanol and dissolved. 0.5 g (4.6 mmol) of 2-amino-5-methyl-pyridine (Aldrich, Wisconsin, USA) was added to the mixture, followed by heating to reflux for 18 hours. After cooling the reaction solution and removing the solvent, the resulting material was extracted with methylene chloride (CH ₂ Cl ₂ ), dried over anhydrous MgSO ₄ and concentrated to obtain a column chromatography (ethyl acetate: n-hexane: methanol = 2: 3: 1), and then recrystallized with ethanol to give a red solid.

Yield: 0.121 g (21%) Melting point: 300 ℃ or more

IR (KBr, cm ^-1 ): 1654 (C = O)

¹ H-NMR (CDCl ₃ ): 10.02 (d, J 1.2 Hz, 1H), 9.57 (d, J 1.2 Hz, 1H), 9.10 (d, J 1.2 Hz, 1H), 7.81 (br. D, J 9.2 Hz, 1H), 7.57 (d, J 9.2 Hz, 1H), 2.46 (s, 3H)

Elemental analysis: C ₁₄ H ₈ N ₄ O ₂ (MW: 264.24)

Calculated Value: C 63.64 H 3.05 N 21.20

Measured value: C 62.87 H 2.55 N 22.34

Example  8) 10- methyl -2,3,6b, 11- Tetraaza - Benzo [a] fluorene -5,6- Dion

0.5 g (2.2 mmol) of 6,7-dichlorophthalazine-5,8-dione and 0.3 g (2.2 mmol) of anhydrous K ₂ CO ₃ were added to 30 mL of anhydrous ethanol and dissolved. 0.5 mL (4.6 mmol) of 2-amino-3-methyl-pyridine (Aldrich, Wisconsin, USA) was added to the mixture, followed by heating to reflux for 16 hours. After cooling the reaction solution and removing the solvent, the resulting material was extracted with methylene chloride (CH ₂ Cl ₂ ), dried over anhydrous MgSO ₄ and concentrated to obtain a column chromatography (ethyl acetate: n-hexane: methanol = 2: 3: 1) and recrystallized again with ethylene acetate and n-hexane to give a purple solid.

Yield: 0.089g (15%) Melting Point: 300 ℃ or higher

IR (KBr, cm ^-1 ): 1655 (C = O)

¹ H-NMR (CDCl ₃ ): 10.12 (d, J 1.2 Hz, 1H), 9.62 (d, J 1.2 Hz, 1H), 9.20 (br. D, J 6.8 Hz, 1H), 7.55 (dd, J 6.8 and 4.4 Hz, 1H), 7.24 (d, J 4.4 Hz, 1H), 2.77 (s, 3H)

Elemental analysis: C ₁₄ H ₈ N ₄ O ₂ (MW: 264.24)

Calculated Value: C 63.64 H 3.05 N 21.20

    Measured value: C 63.74 H 2.67 N 20.78

Example  9) Cytotoxicity Test

Cancer cell lines used for cytotoxicity testing (NCI Institute, USA) are human lung cancer cell line (A549), human ovarian cancer cell line (SK-OV-3), human melanoma cell line (SK-MEL-2), human central nervous system cancer Cell line (XF498), human colon cancer cell line (HCT15), and the cell lines were subjected to SRB (Sulforhodamin B) evaluation.

The cells were seeded in standard 96-well flat bottom microtiter plates and pre-incubated for 24 hours, and the test substances (compounds and comparative materials of Examples 1 to 7) were respectively Diluted from high concentration to low concentration and injected into wells (concentration μg / ml: 10> 1> 0.1> 0.01> 0.001> 0.0001). The treated cells were incubated for 48 hours before the medium was removed from each well and fixed with 10% trichloroacetate at 4 ° C. After washing the trichloroacetate, the treated cells were added to 0.4% SRB solution and incubated for 30 minutes at room temperature. The cells were washed again and then stained by dissolving in 10 ml of bound stain Tris buffer (pH 10.5).

Absorbance of the colored cells was measured at 520 nm using a microtiter plate reader (Molecular Diveces E-max, Sunnyvale, USA). Doxorubicin and etoposide were used as comparative substances, and ED ₅₀ of the test substance and the comparative substance was used. (Μg / mL) values are shown in Table 1 below.

Test substance A549 SK-OV-3 SK-MEL-2 XF 498 HCT 15 Doxorubicin 0.093 0.21 0.11 0.187 0.19 Etoposide 0.577 1.84 1.37 1.43 0.97 Example 1 0.054 0.073 0.11 0.033 0.044 Example 2 0.23 1.04 1.08 0.2 0.71 Example 3 0.59 0.55 0.65 0.45 0.99 Example 4 0.12 0.061 0.034 0.081 0.077 Example 5 0.051 0.02 0.032 0.03 0.047 Example 6 0.022 0.20 0.12 0.12 0.10 Example 7 3.54 1.01 1.77 1.77 0.92

Since the benzofluorene dione compound of the present invention has an excellent cytotoxic effect as compared to conventional anticancer substances, the pharmaceutical composition containing the compound of the present invention or salts thereof has various cancer diseases, especially lung cancer, ovarian cancer and melanoma cancer. It may be used as a therapeutic or prophylactic agent for the treatment of central nervous system cancer, or colon cancer.

Claims (4)

A compound represented by the formula (1) or a pharmaceutically acceptable salt thereof. [Formula 1]

In the above formula, A ring

When ring B is

,

, or

Ring A

When ring B is

Is, In the display of the A ring and the B ring, the side indicated by the broken line represents the side fused to the quinone ring and the imidazole ring in the structure of Chemical Formula 1, respectively. The compound of Chemical Formula 2 and the compound of Chemical Formula 3 are reacted in at least one alcohol solvent selected from methyl alcohol, ethyl alcohol, propyl alcohol and butyl alcohol by using anhydrous potassium carbonate (K ₂ CO ₃ ) as a catalyst. How to manufacture. [Formula 1]

[Formula 2]

[Formula 3]

In the above formula, A ring

When ring B is

,

, or

Ring A

When ring B is

Is, In the display of the A ring and the B ring, the side indicated by the broken line represents the side fused to the quinone ring and the imidazole ring in the structure of Chemical Formula 1, respectively. An anticancer pharmaceutical composition comprising the compound according to claim 1 or a salt thereof as an active ingredient. The pharmaceutical composition according to claim 3, which is used as a prophylactic, therapeutic or therapeutic aid for at least one cancer disease selected from the group consisting of lung cancer, ovarian cancer, melanoma cancer, central nervous system cancer and colon cancer.