KR20090071679A - Pharmaceutical composition for prevention and treatment of cancer disease comprising benzamide derivatives - Google Patents

Abstract

A pharmaceutical composition for preventing and treating cancer which contains a benzamide derivative is provided to prevent and treat various cancer including colorectal cancer by relating to the growth delay of tumor cell and inhibition of cell proliferation. A pharmaceutical composition for preventing and treating cancer comprises a benzamide derivative of the chemical formula 1 or its pharmaceutically allowable salt. In the chemical formula 1, A is -(CH2)0- or -(NR3)-; B is -(CH2)0- or -(NH)n(NR4)-; n is 0-1; R3 and R4 is hydrogen or alkyl each. The benzamide is denoted by the chemical formulas 2 to 5. The pharmaceutical composition is used for preventing and treating colorectal cancer, breast cancer, liver cancer, prostate cancer, ovarian cancer, and gastric cancer. The pharmaceutical composition is used in a form of oral formulation, parenteral formulation, or percutaneous formulation.

Description

Pharmaceutical Composition for Prevention and Treatment of Cancer Disease Comprising Benzamide Derivatives

The present invention relates to a pharmaceutical composition for the prevention and treatment of cancer diseases, and more particularly to the prevention of cancer, in particular colon cancer disease containing a benzamide derivative or a pharmaceutically acceptable salt thereof as an active ingredient and It relates to a pharmaceutical composition for treatment.

Research into the biological activity of benzamide derivatives has been carried out for many years, and as a result, drugs of various structures are known. Among these compounds, anti-cancer drugs are being developed and many low-molecular compounds are currently on the market (Phieng Siliphaivanh and Paul Harrington, 2007; Neeru Khanna and H.J. Jayaram, 2004).

In the present invention, a group of compounds that are known to be biologically active by screening for compounds containing known benzamide groups and performing experiments related to intracellular signal pathway transmission, which is an area where active research has recently been conducted as a mechanism showing anticancer effects. Excavated.

Cancer disease is a disease defined by uncontrolled proliferation and abnormal cell proliferation, and is the second most intractable disease after an accident. In the United States, approximately 1,399,790 were diagnosed with cancer in 2006, and about 564,830 people, or more than 1,500, died of cancer in the same year (American cancer society, 2006). Cancer diseases are becoming a social and economic problem due to the continuous increase in the incidence rate and the early onset of the onset. The causes of cancer are largely external factors (cigarettes, chemicals, radiation, etc.) and internal factors (genetic mutations, hormones, immune conditions, etc.) and these factors work together or act sequentially to initiate or promote cancer production. It is known. Many genes have been identified that affect cancer induction (carcinogens or cancer suppressor genes), and cancers caused by the accumulation of radical genetic mutations cause abnormal cell proliferation and eventually proliferate to other parts of the body. Will move.

Mutations in a single gene may eventually affect signaling pathways that require the normal role of the gene, leading to errors in the pathway, such as the Wnt / β-catenin signaling pathway. Wnt signaling pathways are directly related to anticancer activity and are recognized as the most common feature of human malignancy. Wnt is a group of highly conserved proteins that are involved in cell differentiation, proliferation, maturation, exercise, and the like. Such proteins have been shown to maintain homeostasis in adult tissues during cell germination and to regulate cell proliferation, morphology and motility (Mark L. Johnson et al., 2006; Janssens N. et al., 2006). . Typical Wnt signal transduction leads to the stabilization of β-catenin protein through the regulation of protein kinase, thereby migrating into the cell nucleus and transcriptional activity. This transcriptional activity is reported to be caused by transcription factors of the Lef1 / Tcf group (Moon RT et al., 2002; Reya T and Clevers H, 2005; Wodarz A and Nusse R, 1998).

In the absence of Wnt signaling, GSK-3β (glycogen-synthase kinase-3β) forms a scaffolding protein complex along with anomatous polyposis coli and Axin proteins, leading to major regulation Causes phosphorylation of the factor β-catenin. Phosphorylated β-catenin is degraded by ubiquitination, resulting in a quantitative decrease in cytoplasm, inhibiting Lef1 / Tcf-mediated transcriptional activity (Hart M et al., 1999; Winston JT et al., 1999). .

On the other hand, in the presence of Wnt signaling, Wnt activates Dvl (Dishevelled), a transducer protein, to inhibit GSK-3β activity. Inhibition of GSK eventually results in the accumulation of β-catenin in the cytoplasm, where the accumulated β-catenin enters the nucleus and binds to several transcription factors and induces the expression of target genes. Representative target genes for Wnt signaling are cancers such as matrix metalloproteinases (MMP) such as MMP2, MMP3, MMP7, and MMP9, cyclin D1, Cox-2, c-myc, c-jun, Fra-1 and VEGFR. Many carcinogens are involved in production and invasion. In addition to the presence of Wnt proteins, mutations in various regulators (eg, APC, Axin, β-catenin, etc.) result in the continuous activation of the Wnt signaling pathway. Cancer, ovarian cancer, and gastric cancer), and abnormal activation of the Wnt signaling pathway has been reported in more than 90% of patients with colorectal cancer (Willert, K., and Nusse, R., 1998, Luu). HH, et al., 2004,).

Recent studies have shown that both in vitro and in vitro monoclonal antibodies or inhibitors of Wnt signaling can be used. In vivo (in vivo ) Inhibition of activation of Wnt signaling has been shown to inhibit cell proliferation or invasion and induce apoptosis of cancer cells (He, B. et al., 2004; You, L. et al., 2004).

Most of the currently used cancer treatments are cytotoxic drugs, and serious side effects are seriously threatening the quality of life of patients and advanced tumors such as recurrence or metastatic cancer disease. The case is turning out to be less than expected. Therefore, there is an urgent need for the development of new drugs that show effective efficacy against advanced cancers and can alleviate side effects through improved cancer cell selective specificity. In response to these demands, EGFR inhibitors and VEGFR inhibitors Avastin, which are new molecular-targeted drugs, are being used together with cytotoxic drugs.

Based on the importance of the Wnt signaling pathway, the Wnt signaling molecule has been recognized as a good target for drug screening for the development of drugs that can treat cancer-related diseases such as breast cancer, stomach cancer, liver cancer and colon cancer. Many attempts have been made to identify possible substances (activators or inhibitors) and develop them as new drugs.

The present inventors earnestly studied in order to solve the above problems, by demonstrating that benzamide derivatives and pharmaceutically acceptable salts thereof that act as antagonists having an effect of inhibiting Wnt / β-catenin signaling have an anticancer effect. The present invention has been reached.

Therefore, an aspect of the present invention for achieving the above object is to provide a pharmaceutical composition having a benzamide derivative or a pharmaceutically acceptable salt thereof as an active ingredient having a prophylactic and therapeutic effect of cancer diseases.

Another aspect of the present invention for achieving the above object is to use a benzamide derivative or a pharmaceutically acceptable salt thereof as an active ingredient, cancer has been shown to exhibit anti-cancer activity by inhibiting Wnt / β-catenin signaling It provides a way to prevent or treat.

The first aspect of the present invention relates to a pharmaceutical composition for the prevention and treatment of cancer diseases having a benzamide derivative represented by the following formula (1) and a pharmaceutically acceptable salt thereof as an active ingredient having an anticancer effect.

[Formula 1]

Wherein A is-(CH ₂ ) ₀ -or-(NR ₃ )-, B is-(CH ₂ ) ₀ -or-(NH) n (NR ₄ )-, n is 0 to 1, Wherein R ₃ and R ₄ are the same or different and are each a group represented by hydrogen or alkyl,

R ₁ is hydrogen, an alkyl group, a cycloalkyl group, an alkyl group substituted with a cycloalkyl group, an alkyl group substituted with an aryl group, an aryl group, an acyl group, amino ) Group, carboxy group, carboalkoxy group, carboxyamino group, carbamoyl group, cyano, halo, hydroxy, nitro ( nitro, thio, alkoxy group, aryloxy group, sulfoxy group, and sulfonamide group,

R ₂ is hydrogen, alkyl, aryl substituted alkyl group, aryl group, halo, alkoxy, carboalkoxy, carbamoyl, cyano, hydroxy , Substituted by nitro, thio substituted aryl group, heteroaryl group, and alkyl, halo, alkoxy, carboalkoxy or aryl group Heteroaryl group.

In one embodiment, the benzamide derivatives include those represented by the following formula (2).

[Formula 2]

Wherein n = 1,

R ₂ and R ₄ are as defined above,

R ₅ is hydrogen, an alkyl group, a cycloalkyl group, an alkyl group substituted with a cycloalkyl group, an alkyl group substituted with aryl, an aryl group, acyl, amino, Carboxy, carboalkoxy, carboxyamino, carbamoyl, cyano, halo, hydroxy, nitro, thio ), Alkoxy, aryloxy and sulfoxy.

In one embodiment, the benzamide derivatives include those represented by the following formula (3).

[Formula 3]

Wherein n = 1,

R ₂ and R ₄ are as defined above,

R ₆ and R ₇ are the same or different and each is hydrogen, an alkyl group, an alkyl group substituted with alkynyl or alkoxy, an alkyl group substituted with cycloalkyl or aryl, morpholine, alkyl Substituted morpholine groups, piperazine and piperazine groups substituted with alkyl, alkenyl, alkynyl, carboxyalkoxy or carbamoyl.

In one embodiment, the benzamide derivatives include those represented by the following formula (4).

[Formula 4]

Wherein n = 0,

R ₁ , R ₂ and R ₄ are as defined above.

In one embodiment, the benzamide derivatives include those represented by the following formula (5).

[Formula 5]

Wherein R ₁ and R ₃ are as defined above,

R ₈ is alkyl, alkyl group substituted by aryl group, aryl, halo, alkyl, alkoxy, carboalkoxy, carbamoyl, cyano , Aryl, heteroaryl, alkyl, halo, alkoxy, carboalkoxy or aryl groups substituted by hydroxy, nitro or thio It is selected from a heteroaryl group substituted with.

The pharmaceutical composition of the present invention is involved in cell growth inhibition and tumor growth retardation of tumor cells, including the inhibitory effect of Wnt signaling pathways, and various cancer diseases including colorectal cancer (CRC). Effective in the prevention and treatment of them.

Prior to describing the contents of the present invention in detail, some terms used herein are defined as follows.

a) alkyl group, consisting of 1 to 10 carbon atoms, including linear or branched saturated or unsaturated hydrocarbons. Wherein at least one hydrogen is acyl, amino, carboalkoxy, carboxy, carboxyamino, -O-carbamoyl (-O- (C = O) -NH ₂ ), Cyano, halo, hydroxy, nitro, thio, alkyl, cycloalkyl, aryl, alkoxy ), Aryloxy, sulfoxy, and guanido may be substituted with one or more substituents selected from the group consisting of up to the maximum number that can be substituted regardless of order and type.

b) cycloalkyl groups: 3--12 pentagonal ring structures containing saturated or partially unsaturated hydrocarbons, containing 0-5 heteroatoms such as oxygen, sulfur, nitrogen, etc. The ring may be a single ring or fused ring compound of 3 to 12 hexagons. Wherein at least one hydrogen is acyl, amino, carboalkoxy, carboxy, carboxyamino, -O-carbamoyl (-O- (C = O) -NH ₂ ) , Cyano, halo, hydroxy, nitro, thio, alkyl, cycloalkyl, aryl, alkoxy ), Aryloxy, sulfoxy, and guanido may be substituted with one or more substituents selected from the group consisting of up to the maximum number that can be substituted regardless of order and type.

Specific examples of the cycloalkyl group include cyclopropyl, cyclobutyl, cyclopentyl, cyclohehexyl, cycloheptyl, cyclooctyl, morpholinyl , Homomorpholinyl, thiomorpholinyl, homothiomorpholinyl, thiomorpholinyl S-oxide, thiomorpholinyl S, S-dioxide ( thiomorpholinyl S, S-dioxide, piperidinyl, homopiperidinyl, piperazinyl, homopiperazinyl, pyrrolidinyl, pyrrolinyl Tetrahydropyranyl, tetrahydrofuranyl, tetrahydrofuranyl, tetrahydrothienyl, oxazolidinonyl, dihydropyrazolyl, dihydropyranolyl, dihydropyrazolyl Dihydropyrrolyl, dihydropyrazinyl, dihydropyridinyl, dihydropyrimidinyl, dihydrofuryl, dihydropyranyl, and the like. have.

c) aryl group: Aromatic groups consisting of 5 to 15 angular unsaturated hydrocarbons in simple or fused rings and heteroatoms such as oxygen, sulfur and nitrogen. It includes all of the included heteroaromatic groups. Wherein at least one hydrogen is acyl, amino, carboalkoxy, carboxy, carboxyamino, -O-carbamoyl (-O- (C = O) -NH ₂ ), Cyano, halo, hydroxy, nitro, thio, alkyl, cycloalkyl, aryl, alkoxy ), Aryloxy, sulfoxy, and guanido may be substituted with one or more substituents selected from the group consisting of up to the maximum number that can be substituted regardless of order and type.

Specific examples of the aryl group include phenyl, 1-naphtyl, 2-naphtyl, 2-naphtyl, pyridinyl, pyrimidinyl, quinolinyl, Benzothienyl, indolyl, pyrazinyl, isoindolyl, isoquinolyl, quinazolinyl, quinoxalinyl, phthalazinyl (phthalazinyl), imidazolinyl, isoxazolyl, pyrazolyl, oxazolyl, thiazolyl, indolizinyl, indazolyl ), Benzothiazolyl, benzimidazolyl, benzofuranyl, benzofuranyl, thienyl, pyrrolyl, oxadiazolyl, thiadiazolyl thiadiazolyl, triazolyl, tetrazolyl, oxazolopyridinyl, imidazopyridrid inyl, isothiazolyl, cinolinyl, carbazolyl, isochromanyl, chromanyl, tetrahydroisoquinolinyl, isodololinyl (isoindolinyl), isobenzotetrahydrofuranyl, isobenzotetrahydrothienyl, isobenzothienyl, benzoxazolyl, pyridopyridinyl, Benzotetrahydrofuranyl, benzotetrahydrothienyl, purinyl, benzodioxolyl, triazinyl, phenoxazinyl, phenoxazinyl (phenothiazinyl), pterididinyl, benzothiazolyl, imidazopyridinyl, imidazothiazolyl, daihai Robbenz isoxazinyl, benzisoxazinyl, benzoxazinyl, benzoxazinyl, dihydrobenzisothiopyranyl, benzopyranyl, benzothiopyranyl, benzothiopyranyl Coumarinyl, isocoumarinyl, chromonyl, chromonyl, chromanonyl, pyridinyl-N-oxide, tetrahydroquinolinyl-N-oxide (tetrahydroquinolinyl- N-oxide, dihydroquinolinyl, dihydroquinolinonyl, dihydroisoquinolinonyl, dihydrocoumarinyl, dihydroisocoumarinyl, dihydroisocoumarinyl, isoin Isolinindolinonyl, benzodioxanyl, benzoxazolinonyl, pyrrolyl-N-oxide, pyrimidinyl- Pyrimidinyl-N-oxide, pyrazinyl-N-oxide, quinolinyl-N-oxide, indolyl-N-oxide oxide, indolinyl-N-oxide, pyrazinyl-N-oxide, isoquinolyl-N-oxide, kunazolinyl- N-oxide (quinazolinyl-N-oxide), quinoxalinyl-N-oxide, phthalazinyl-N-oxide, imidazolinyl-N-oxide ( imidazolinyl-N-oxide, isoxazolyl-N-oxide, oxazolyl-N-oxide, thiazolyl-N-oxide , Indolizinyl-N-oxide, indazolyl-N-oxide, benzothiazolyl-N-oxide, benzimidazolyl -Benzimidazolyl-N-oxide, pyrrolyl-N-oxide, oxadiazolyl-N-oxa Oxadiazolyl-N-oxide, thiadiazolyl-N-oxide, triazolyl-N-oxide, tetrazolyl-N-oxide (tetrazolyl-N -oxide) and the like.

d) halo: generically fluoro, chloro, bromo and iodo.

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

Benzamide derivatives according to the present invention include not only the compounds represented by Chemical Formulas 1 to 5, but also pharmaceutically acceptable acid or base addition salts thereof, and stereochemically isomeric forms thereof. Any salt that maintains the activity of the parent compound and does not cause an undesirable effect is included and is not particularly limited. Such salts include inorganic salts and organic salts, and preferably salts of the following acids. More specifically, acetic acid, nitric acid, aspartic acid, sulfonic acid, sulfonic acid, maleic acid, maleic acid, glutamic acid, formic acid, Succinic, phosphoric, phthalic, tannic, tartaric, hydrobromic, propionic, benzenesulfonic, benzoic acid ), Stearic acid, esyl acid, esyl acid, lactic acid, bicarbonic acid, bisulfuric acid, busulfuric acid, bitartaric acid, oxalic acid, butyric acid ( butyric, calcium edentate, kimsylic acid, camsylic, carbonic acid, carbonic acid, chlorobenzoic acid, citric acid, citric acid, edetic acid, toluenesulfonic acid, and edisonin Acid (edisylic), esylic acid (esylic), fumaric acid (fumaric), gluceptic acid (gluceptic), pamoic acid, gluconic acid, glycolyl Glycollylarsanilic, methylnitric acid, polygalactronic chloric acid, hydrochloric, hydroiodic, hydroxynaphthoic, isethionic, lactobionic Acid (lactobionic), mandelic acid (madelic), estolic acid (estolic), methylsulfuric acid (methylsulfuric), mumic acid, muconic acid, para-nitromethanesulfonic acid (p- nitromethanesulfonic, hexamic, pantothenic, monohydrogen phosphoric, dihydrogen phosphoric, salicylic, sulfamic, sulfanilic acid, sulfanilic acid, Methanesulfonic acid, theolic acid, and the like.

Also in the form of the basic salts are, for example, ammonium salts, alkali and alkaline earth metal salts, in particular, the salts with organic, base, sodium, potassium, magnesium and calcium salts, for example bezatin, N-methyl-D -Glucamine, hydrabamine salts, and salts with amino acids such as arginine, lysine.

In contrast, the salt form may be converted to the free form by treatment with a suitable base or acid.

In one embodiment, the preferred salt of the benzamide derivative of the present invention is benzo [1,3] -dioxol-5-carboxylic acid [4- (4-methoxy-phenylsulfamoyl) -phenyl] -amide.

In addition, the present invention using the benzamide derivative represented by any one of the formulas (1) to 5 and its pharmaceutically acceptable salts as an active ingredient, has the effect of inhibiting Wnt / β-catenin signaling to exhibit anticancer activity By providing a method for preventing or treating cancer diseases.

The pharmaceutical composition of the present invention may be usefully used for the prevention and treatment of various cancer diseases, and the cancer diseases include, but are not limited to, colorectal cancer, breast cancer, liver cancer, prostate cancer, ovarian cancer, stomach cancer, and the like. have. Preferably the cancer disease is colorectal cancer.

Preparation of the compound

The compounds of the present invention can be prepared by various synthetic or semisynthetic techniques. Suitable starting materials may be apparent to those skilled in the art, or may be prepared by known methods, or such starting materials may be purchased commercially. Those skilled in the art will appreciate that the synthetic route may be modified to use other starting materials or selective reagents, and that there may be appropriate changes in the reaction conditions (eg, temperature, solvent, etc.) to obtain the desired modifications. In addition, those skilled in the art will recognize that protecting groups may be required for the preparation of certain compounds and will know the reaction conditions suitable for the selected protecting group. Accordingly, the exemplary methods and embodiments described herein are illustrative of the invention and are not to be construed as limiting the scope of the invention.

Next, as an example, a method for preparing the benzamide derivative compound represented by Chemical Formula 4 will be described in detail with reference to Scheme 1 below.

Scheme 1

N is 0 or 1,

R ₁ is hydrogen, an alkyl group, a cycloalkyl group, an alkyl group substituted with a cycloalkyl group, an alkyl group substituted with an aryl group, an aryl group, an acyl group, amino ) Group, carboxy group, carboalkoxy group, carboxyamino group, carbamoyl group, cyano, halo, hydroxy, nitro ( nitro, thio, alkoxy group, aryloxy group, sulfoxy group, and sulfonamide group,

R ₂ is hydrogen, alkyl, aryl substituted alkyl group, aryl group, halo, alkoxy, carboalkoxy, carbamoyl, cyano, hydroxy , Substituted by nitro, thio substituted aryl group, heteroaryl group, and alkyl, halo, alkoxy, carboalkoxy or aryl group Heteroaryl group selected from the group consisting of

R ₄ is a group represented by hydrogen or alkyl, and

X is a group activated with halo, methansulfonyl, phenylsulfonyl, alkoxy and the like.

Scheme 1 shows a coupling reaction for obtaining the compound represented by Chemical Formula 4, specifically, the intermediate ( 1 ) activated with alkoxy is refluxed under the intermediate ( 2 ) and an ethanol solvent. By stirring, a benzamide derivative compound of Chemical Formula 4 may be obtained.

In addition, a benzamide derivative compound of Chemical Formulas 2 to 4 may be obtained by a coupling reaction as in Scheme 1.

In addition, a method for preparing the benzamide derivative compound represented by Chemical Formula 5 will be described in detail with reference to Scheme 2 below.

Scheme 2

R ₁ is as defined above,

R ₃ is a group represented by hydrogen or alkyl,

R ₈ is alkyl, alkyl group substituted by aryl group, aryl, halo, alkyl, alkoxy, carboalkoxy, carbamoyl, cyano , Aryl, heteroaryl, alkyl, halo, alkoxy, carboalkoxy or aryl groups substituted by hydroxy, nitro or thio And is selected from a heteroaryl group substituted with

X is a group activated with halo, methansulfonyl, phenylsulfonyl, alkoxy and the like.

Scheme 2 shows a coupling reaction for obtaining the compound represented by Chemical Formula 5, specifically, the activated intermediate (4) and the amine intermediate (3) are reflux stirred in an acetonitrile solvent as an example. Benzamide derivative compounds of formula (5) can be obtained.

Pharmaceutical Compositions and Administration

In another aspect of the invention, a composition suitable for pharmaceutical use is provided comprising one or more compounds of the invention and a pharmaceutically acceptable carrier, excipient or diluent.

The term "composition" as used herein, is intended to include outputs comprising specific components (and, if specified, specific amounts), as well as any output obtained directly or indirectly from a combination of specific components in a particular amount. do. "Pharmaceutically acceptable" means that the carrier or excipient is suitable for the other ingredients of the formulation and is not harmful to their receptors.

In preparing the pharmaceutical composition, the carrier is selected according to the formulation to be prepared and can be formulated by mixing it with the active ingredient of the benzamide derivative in an appropriate ratio.

The pharmaceutical composition of the present invention may be administered parenterally, topically, orally or locally for therapeutic treatment. For example, various aqueous carriers may be used, such as water, buffered water, 0.04% saline, 0.3% glycol, and may include other proteins that enhance stability, such as albumin, lipoproteins, globulins, and the like. As a result, the composition can be sterilized by known sterilization techniques. The solution may be packaged for use, filtered and lyophilized under aseptic conditions, and the freeze-dried preparation is combined with the sterilized solution prior to administration.

Oral compositions generally include an inert diluent or an edible carrier. They are enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound may be included with excipients and used in the form of tablets, troches or capsules.

When preparing the oral composition, a conventional pharmaceutical carrier can be used. For example, in the case of oral liquid preparations such as suspensions, syrups, elixirs and solutions, water, glycols, oils, alcohols and the like may be used as carriers, and in the case of solid preparations such as powders, pills, capsules and tablets Starch, sugar, kaolin, lubricants, binders, disintegrants and the like can be used. Given the ease of administration, tablets and capsules are the most convenient dosage forms, and in the case of tablets and pills, it is more preferable to prepare them as enteric preparations.

In the case of parenteral preparations, sterile water is usually used as a carrier, and other ingredients such as dissolution aids may also be included.

Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions can be prepared using suitable dispersants, wetting agents or suspending agents according to known techniques. Solvents that can be used here include water, Ringer's solution and isotonic NaCl solution, and sterile fixed oils are also commonly used as solvents or suspending media. Any non-irritating fixed oil may be used for this purpose, including mono or diglycerides, and other fatty acids such as oleic acid may also be used in the preparation of injectables.

Therapeutically effective amount of the pharmaceutical composition of the invention in known treatment for a disease that requires a biometric (in vivo ) and in vitro vitro) may be determined empirically by experiment the compound in a model system. The therapeutically effective amount means the amount of active ingredient effective to alleviate or reduce the symptoms of a disease in need of treatment or to reduce or delay the onset of a clinical marker or condition of a disease in need of prevention.

When administering the active ingredient of the composition according to the invention, specifically the benzamide derivative of any one of formulas 1-5, for clinical purposes, the total daily dose to be administered to the patient in a single dose or in separate doses is 0.1 per kg of body weight. Although the range of ˜100 mg is more preferred, the specific dose level for a particular patient is determined by the specific compound to be used, the weight of the patient, sex, health condition, diet, the time of administration of the drug, the method of administration, the rate of excretion, the mixing of the drug and the severity of the disease. Can be changed accordingly.

In some cases, the benzamide derivative may be used to formulate an effective pharmaceutical composition in the form of its prodrug or the like.

In addition, the composition according to the present invention may further include other ingredients that do not inhibit the action or assist in the action of the active ingredient, it may be formulated in various forms known in the art. The composition according to the invention may preferably further contain a known anticancer agent.

Hereinafter, the content of the present invention will be described in more detail with reference to the following examples, which are only intended to help the understanding of the present invention and are not intended to limit the scope of the present invention.

Compound 1: 4- [N '(4,6-dimethyl-benzothiazol-2-yl) -hydrazinocarbonyl] -benzoic acid methyl ester

The compound was purchased from Chemdiv (Cat No. C502-0485) and used.

Compound 2: 4-fluoro-benzoic acid N ' (4,7-dimethoxy-benzothiazol-2-yl) -hydrazide

The compound was purchased from Chemdiv (Cat No. C502-0633) and used.

Compound 3: 4- {4- [N '-(4-methoxy-7-methyl-benzothiazol-2-yl) -hydrazinocarbonyl] -benzenesulfonyl} -piperazine-1-carboxylic acid ethyl Ester

The compound was purchased from Chemdiv (Cat No. C502-0571) and used.

Compound 4: 4- (2,6-dimethyl-morpholin-4-sulfonyl) -N- [5- (4-fluoro-phenyl)-[1,3,4] oxadiazol-2-yl ] -Benzamide

The compound was purchased from Chemdiv (Cat No. C079-0047) and used.

Compound 5: N- (4-benzo [1,3] dioxol-5-yl-thiazol-2-yl) -4- (3,5-dimethyl-piperidine-1-sulfonyl) -benz amides

The compound was purchased from Chemdiv (Cat No. 3331-4726) and used.

Compound 6: Benzo [1,3] -dioxol-5-carboxyl acid [4- (4-methoxy-phenylsulfamoyl) -phenyl] -amide

The compound was purchased from Chemdiv (Cat No. 2884-3651) and used.

The above compounds were tested for anticancer effects by the following method.

Experimental Example 1: Effect of Wnt / β-catenin signaling activity efficacy using cell line

a) Measurement method of IC ₅₀

end. Experiment method

In order to confirm the activity of Wnt / β-catenin signaling in HEK293 cells, a binding site of Tcf / Lef transcriptional regulatory protein to which five β-catenin is bound, and a firefly luciferase of the marker protein Drosophila expressed under its control The plasmid was introduced into the cell to prepare a transformed cell line (PCT / KR2006 / 005837). Two days before the compound treatment, 2 x 10 ⁴ transformed cells were plated on a 96 well plate, mixed with Wnt-conditioning medium and general culture medium 1: 4, and then sequentially diluted (50.0, 16.7, 5.6, 1.9, 0.6, 0.2 uM) were simultaneously treated and incubated for 18 hours to determine the expression level of the label gene (luciferase) using the Steady-Glo Luciferase Kit (Promega). The Wnt-conditioning medium is a culture medium obtained by culturing the cell line CRL2647 (ATCC) expressing Wnt3a for two days and then collecting the medium and filtration. At this time, the negative control used was normal culture solution only, the positive control was used to treat only Wnt-conditioned medium. The inhibition rate of Wnt signaling was calculated as follows.

[Equation 1]

Inhibitory percentage = (Ct-Cn) / (Cp-Cn) * 100

(Ct: test group, Cn: control medium treatment group, Cp: Wnt-conditioned medium treatment group)

I. Experiment result

Table 1. Inhibitory Effects of Wnt Signaling

compound Compound 1 Compound 2 Compound 3 Compound 4 Compound 5 Compound6 IC ₅₀ (HEK293, μM) 11.5 21.5 6.8 2.41 2.78 14.5

As a result of measuring the activity of the Wnt signaling pathway compared to the positive control treated with Wnt-conditioned medium only, Compounds 3, 4 and 5 showed a 50% reduction in 6.8, 2.41 and 2.78 μM concentrations, respectively. Nos. 1 and 6 showed 50% reduction at 11.5 and 14.5 μM. Compound 2 showed a reduction effect at 21.5 μM.

b) TOP-Flash / FOP-Flash Reporter Assay

end. Experiment method

To investigate the effects of selectively inhibiting the Wnt signaling pathway, a reporter plasmid (TOP-Flash) containing a Tcf-4 binding site or a mutant Tcf-4 binding site was included in an HCT116 cell line in which the Wnt signaling pathway was activated. 0.4 μg of reporter plasmid (FOP-Flash) (Korinek V. et al, 1997) and 0.1 μg of Renilla Luciferase reporter plasmid pRLTK (promega) capable of correcting transfection were transfected using Lipofectamin 2000 (Invitrogen). After that, the cells were incubated for 18 hours. Then, after 18 hours of treatment with the compound, the expression change of the marker gene by the Tcf-4 promoter was examined using the Dual-Glo Luciferase Kit (Promega).

I. Experiment result

The results of the experiment are shown in FIG. As can be seen from FIG. 1, after expressing the two plasmids temporarily and comparing the expression of the untreated TOP-flash to 100%, the concentration-dependent TOP-flash in the compounds 3 and 4 was compared. On the other hand, when the FOP-flash plasmid was injected into the cells, the expression reduction effect by the compound was not seen.

Experimental Example 2: Inhibition of Cell Proliferation

end. Experiment method

In order to confirm the effect of inhibiting cell growth on HCT116 cells and SW480 cells, 1 × 10 ⁴ were dispensed into 96 well plates, and then cultured for 24 hours. CellTiter-Glo Cell Viability Assay Kit was prepared by diluting the compounds sequentially, treating them with 6 concentrations (50.0, 16.7, 5.6, 1.9, 0.6, 0.2 uM) and GI ₅₀ , which inhibits cell growth by 50% after 48 hours. It was determined using (Promega). GI ₅₀ The following formula was used to determine the value.

[Equation 2]

GI ₅₀ = [(Ti-Tz)] / (C-Tz)] * 100 (if, Ti≥Tz)

GI ₅₀ = [(Ti-Tz)] / Tz * 100 (if, Ti <Tz)

Time zero: Tz, control: C, test group: Ti

I. Experiment result

Table 2. Effect of Cell Proliferation Inhibition

compound Compound 1 Compound 2 Compound 3 Compound 4 Compound 5 Compound6 GI ₅₀ (HCT116, μM) 28.8 19.5 4.3 4.1 13.4 16.6 GI ₅₀ (SW480, μM) ND ND 9.8 ND 12 ND

ND: not determined

In HCT116 cells, compounds 3 and 4 showed 50% cell proliferation reduction at 4.3μM and 4.1μM concentrations, and compounds 2, 5 and 6 showed 50% reduction at 19.5μM, 13.4μM and 16.6μM, respectively. Showed. Compound 1 showed a 50% reduction at 28.8 μM.

In SW480 cells, compound 3 and 5 showed 50% cell proliferation reduction at concentrations of 9.8 μM and 12 μM.

Experimental Example 3: Tumor regression assay

end. Experiment method

Balb / c nude mice (15-16 g, 6 weeks old, females) were prepared and purified in a sterile animal room for at least one week before being used for the experiment. HCT 116 Cancer Cell Lines by an after induce axillary tumor by subcutaneous injection in the upper gyeongapbu between the chest wall of the right side by 0.3 ml per mouse, with an average tumor size of about 50-60 mm emitter ³ reaches bitter discloses a test substance is administered the last day ( day 20) Administration was repeated at a dose of 50 mg / 10ml / kg once daily, intraperitoneally, until the day before. Irinotecan, used as a positive control, was administered intravenously (iv) once a week at 50 mg / 10 ml / kg. Animal weight changes were measured three times per week during the study to determine the degree of toxicity, such as weight loss due to drug administration. Tumor size was discarded and measured in three directions using a caliper (verier caliper) and then expressed by the following equation.

[Equation 3]

Tumor volume = (length × width × height) / 2

I. Experiment result

Observation of the general symptoms and body weight of the animals during the administration of HCT 116 cancer cell transplantation to determine the degree of toxicity of repeated compound intraperitoneal administration at a dose of 50 mg / 10ml / kg for 20 days (once per day) It was. The compound showed no general symptoms and statistically significant weight loss due to drug administration during the administration period. In the 50 mg / kg Irinotecan-treated group, the results at day 19 (three animals died in the last day's solvent control group and the results are the last 19 days) were 15% (p <0.005) compared to the solvent control group. Statistically significant weight loss was observed (Table 3, Figure 2).

Table 3. Changes in body weight and tumor size after drug administration

compound Tumor size (mm ³ ) Tumor size reduction (%) Weight loss (%) Compound 3 (50mpk, ip) 56 28.6 1.7 Irinotecan (50mpk, iv) 57 73.4 15.4

Mean tumor size changes after transplantation of HCT 116 cancer cell lines are shown in Table 3 and FIG. 3. Results 19 days after drug administration (three animals died in the last control group, the result is the last 19 days) showed 28.6% (p <0.5 vs. Control) tumor growth inhibition effect compared to the solvent control group In the positive control (Irinotecan) group, 73.4% (p <0.005 vs. Control) showed statistically significant tumor growth inhibitory effect (Table 3, Figure 3).

In the case of repeated intraperitoneal administration at 50 mg / kg in human rectal cancer cell (HCT-116) transplanted nude mice, no unusual general symptoms and statistically significant weight loss were observed during the administration period. This may be due to the growth of the tumor rather than to

Compound 3 was confirmed to have the effect of effectively inhibiting tumor growth in the anti-cancer model without weight change.

1 shows the effect of the pharmaceutical composition of the present invention on Tcf-4 activity,

Figure 2 shows the weight change after administration of the pharmaceutical composition of the present invention, and

Figure 3 shows the tumor size change after administration of the pharmaceutical composition of the present invention.

Claims (8)

Benzamide derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof is contained as an active ingredient cancer prevention and treatment pharmaceutical composition having an anticancer effect: [Formula 1]

Wherein A is-(CH ₂ ) ₀ -or-(NR ₃ )-, B is-(CH ₂ ) ₀ -or-(NH) n (NR ₄ )-, n is 0 to 1, Wherein R ₃ and R ₄ are the same or different and are each a group represented by hydrogen or alkyl, R ₁ is hydrogen, an alkyl group, a cycloalkyl group, an alkyl group substituted with a cycloalkyl group, an alkyl group substituted with an aryl group, an aryl group, an acyl group, amino ) Group, carboxy group, carboalkoxy group, carboxyamino group, carbamoyl group, cyano, halo, hydroxyl, nitro ( nitro, thio, alkoxy groups, aryloxy groups, sulfoxy groups, and sulfonamide groups, and R ₂ is hydrogen, alkyl, aryl substituted alkyl group, aryl group, halo, alkoxy, carboalkoxy, carbamoyl, cyano, hydroxy Aryl groups, heteroaryl groups, and alkyl, halo, alkoxy, carboalkoxy or aryl groups substituted with oxy, nitro, thio It is selected from a substituted heteroaryl group. The pharmaceutical composition for preventing and treating cancer according to claim 1, wherein the benzamide derivative is represented by the following Chemical Formula 2: [Formula 2] Wherein n = 1, R ₂ and R ₄ are as defined in claim 1, and R ₅ is hydrogen, an alkyl group, a cycloalkyl group, an alkyl group substituted with a cycloalkyl group, an alkyl group substituted with aryl, an aryl group, acyl, amino, Carboxy, carboalkoxy, carboxyamino, carbamoyl, cyano, halo, hydroxy, nitro, thio ), Alkoxy, aryloxy and sulfoxy. The pharmaceutical composition for preventing and treating cancer according to claim 1, wherein the benzamide derivative is represented by the following Chemical Formula 3: [Formula 3]

Wherein n = 1, R ₂ and R ₄ are as defined in claim 1, and R ₆ and R ₇ are the same or different and each is hydrogen, an alkyl group, an alkyl group substituted with alkynyl or alkoxy, an alkyl group substituted with cycloalkyl or aryl, morpholine, alkyl Substituted morpholine groups, piperazine and piperazine groups substituted with alkyl, alkenyl, alkynyl, carboxyalkoxy or carbamoyl. According to claim 1, wherein the benzamide derivative is a pharmaceutical composition for preventing and treating cancer, characterized in that represented by the following formula (4):  [Formula 4]

Wherein n = 0, and R ₁ , R ₂ and R ₄ are as defined in claim 1. According to claim 1, wherein the benzamide derivative is a pharmaceutical composition for preventing and treating cancer, characterized in that represented by the following formula (5): [Formula 5]

Wherein R ₁ and R ₃ are as defined in claim 1, and R ₈ is alkyl, alkyl group substituted by aryl group, aryl, halo, alkyl, alkoxy, carboalkoxy, carbamoyl, cyano , Aryl, heteroaryl, alkyl, halo, alkoxy, carboalkoxy or aryl groups substituted by hydroxy, nitro or thio It is selected from the heteroaryl group substituted by. The method according to any one of claims 1 to 5, wherein the pharmaceutical composition is used for the prevention and treatment of cancer diseases selected from the group consisting of colorectal cancer, breast cancer, liver cancer, prostate cancer, ovarian cancer and gastric cancer. Pharmaceutical composition for preventing and treating cancer. The pharmaceutical composition according to any one of claims 1 to 5, characterized in that the pharmaceutical composition comprises a pharmaceutically acceptable carrier suitable for formulation into oral, parenteral, injectable or transdermal formulations. Pharmaceutical compositions. A method of preventing or treating cancer by administering a composition according to any one of claims 1 to 5 to a patient to inhibit Wnt / β-catenin signaling and thereby exhibit anticancer activity.

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