KR100983319B1 - Use in anti-cancer agent of [meso-Bis(o-hydroxyphenyl)bis(p-nitro)salen, 2,2'-(1E,1'E)-((1R,2S)-1,2-bis(2-hydroxyphenyl)ethane-1,2-diyl)bis(azan-1-yl-1-ylidene)bis(methan-1-yl-1-ylidene)bis(4-nitrophenol)] - Google Patents

Abstract

The present invention provides meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen [meso-Bis (o-hydroxyphenyl) bis (p-nitro) salen, 2,2 '-(1E, 1'E)- ((1R, 2S) -1,2-bis (2-hydroxyphenyl) ethane-1,2-diyl) bis (azan-1-yl-1-ylidene) bis (methan-1-yl-1-ylidene) bis (4-nitrophenol)] The present invention relates to the use of a compound as an anticancer agent, and more particularly, meso-bis (ortho-) of Formula 1, which inhibits abnormal cell division and proliferation by reducing the expression level of Cyclin D1 protein. It relates to a pharmaceutical composition for the prevention and treatment of cancer diseases comprising a hydroxyphenyl) bis (para-nitro) salen compound and the compound or a pharmaceutically acceptable salt thereof.

[Formula 1]

Since meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen of the present invention has an effect of reducing the expression of cyclin D1 protein and inducing cell cycle progression, it inhibits the proliferation of cancer cells and prevents cell death. Induction can be useful as a pharmaceutical composition for the prevention and treatment of cancer.

Anticancer agent, cyclin D1, cell cycle progression inhibition, meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen

Description

Use in anti-cancer agent of [meso-Bis (o-hydroxyphenyl) bis (p-nitro) salen, 2,2 ' -(1E, 1'E)-((1R, 2S) -1,2-bis (2-hydroxyphenyl) ethane-1,2-diyl) bis (azan-1-yl-1-ylidene) bis (methan- 1-yl-1-ylidene) bis (4-nitrophenol)]}

The present invention provides meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen [meso-Bis (o-hydroxyphenyl) bis (p-nitro) salen, 2,2 '-(1E, 1'E)- ((1R, 2S) -1,2-bis (2-hydroxyphenyl) ethane-1,2-diyl) bis (azan-1-yl-1-ylidene) bis (methan-1-yl-1-ylidene) bis (4-nitrophenol)] The present invention relates to the use of a compound as an anticancer agent, and more particularly, meso-bis (ortho-) of Formula 1, which inhibits abnormal cell division and proliferation by reducing the expression level of Cyclin D1 protein. It relates to a pharmaceutical composition for the prevention and treatment of cancer diseases comprising a hydroxyphenyl) bis (para-nitro) salen compound and the compound or a pharmaceutically acceptable salt thereof.

All cells proliferate and grow normally through cell cycle regulation by various mechanisms.

The cell cycle is composed of two phases, interphase and mitotic phase. The interphase, in turn, is a G1 phase where cells synthesize the various proteins necessary for cell division, an S phase where DNA synthesis occurs, and growth for mitosis. It is composed of three stages such as G2 stage in which preparation is performed, and the cell differentiation process is a mitosis-phase stage.

Cyclin proteins involved in the cell division process regulate the cell cycle stage by forming cyclin / CDK complexes with phosphorylation enzymes called Cyclin Dependent Kinase (CDK). Cyclin D1 is a protein that plays an important role in the progression from G1 to S phase.

When cyclin D1 is complexed with CDK4 or CDK6 and becomes active, phosphorylation of Retinoblastoma protein (pRb) is performed. Phosphorylation by cyclin D1 leads to inactivation and the release of E2F (Seminars Cancer Biol., 6 , 99, 1995).

As a result, the inactivated E2F is activated as a transcription factor, and the process of transcription and translation of proteins for cell cycle progression is smooth, and the cell cycle is shifted from the G1 stage to the S stage. In normal cells, the cell cycle regulation mechanism is normally performed, so that cell division and proliferation are properly performed, and the cell number is properly regulated, whereas in cancer cells, excessive proliferation and growth of cells are known to occur due to abnormal cell cycle progression. In fact, some cancer cells have observed cyclin D1 gene amplification or protein overexpression (Am. J. Pathology, 147 , 545, 1995).

Therefore, if the expression level of cyclin D1 is decreased, phosphorylation of pRb is not normally performed, and thus, the cell cycle is maintained at the G1 stage, thereby preventing abnormal cell division and proliferation of cancer cells and applying it to cancer treatment.

Accordingly, the present inventors have made diligent efforts to find a substance capable of inhibiting the proliferation of cancer cells by reducing the expression level of cyclin D1 protein. As a result, a new salen ligand derivative having stereospecificity regulates the cell cycle. It was confirmed that the amount of expression was reduced and the present invention was completed.

Finally, the present invention provides meso-bis (o-hydroxyphenyl) bis (p-nitro) salen, 2, which reduces the expression level of cyclin D1 protein. 2 '-(1E, 1'E)-((1R, 2S) -1,2-bis (2-hydroxyphenyl) ethane-1,2-diyl) bis (azan-1-yl-1-ylidene) bis ( methan-1-yl-1-ylidene) bis (4-nitrophenol)] and its use as an anticancer agent.

In order to achieve the above object, the present invention is a meso-bis (ortho-hydroxyphenyl) bis (para-nitro) of formula (1) to suppress abnormal cell division and proliferation by reducing the expression level of Cyclin D1 (Cyclin D1) protein Salen, meso-Bis (o-hydroxyphenyl) bis (p-nitro) salen, 2,2 '-(1E, 1'E)-((1R, 2S) -1,2-bis (2-hydroxyphenyl) ethane -1,2-diyl) bis (azan-1-yl-1-ylidene) bis (methan-1-yl-1-ylidene) bis (4-nitrophenol)] compound.

[Formula 1]

The present invention also provides a use of the meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen compound as an anticancer agent.

Specifically, the present invention provides a pharmaceutical composition for the prevention and treatment of cancer diseases, including the meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen or a pharmaceutically acceptable salt thereof.

The cancer disease includes general cancer diseases, preferably gastric cancer, colon cancer, breast cancer, lung cancer, non-small cell lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or ocular melanoma, uterine cancer, ovarian cancer, rectal cancer , Anal leiomyoma, fallopian tube dysfunction, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma Cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocyte lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, One or more diseases selected from brain stem glioma, pituitary adenoma, and the like.

Meso-bis (o-hydroxyphenyl) bis (p-nitro) salen, 2,2 '-(1E, 1'E)-of the present invention ((1R, 2S) -1,2-bis (2-hydroxyphenyl) ethane-1,2-diyl) bis (azan-1-yl-1-ylidene) bis (methan-1-yl-1-ylidene) bis (4-nitrophenol)] is a pharmaceutical composition for preventing and treating cancer by inhibiting the proliferation of cancer cells and inducing apoptosis because it has an effect of reducing the expression level of cyclin D1 (Cycling D1) to induce abnormal cell cycle progression. It can be usefully used.

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

The present invention is a meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen [meso-Bis (Formula 1) of inhibiting abnormal cell division and proliferation by reducing the expression level of Cyclin D1 (Cyclin D1) protein o-hydroxyphenyl) bis (p-nitro) salen, 2,2 '-(1E, 1'E)-((1R, 2S) -1,2-bis (2-hydroxyphenyl) ethane-1,2-diyl) bis (azan-1-yl-1-ylidene) bis (methan-1-yl-1-ylidene) bis (4-nitrophenol)] compound.

[Formula 1]

In the present invention, the meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen of the formula (1) can be prepared by a synthesis method well known in the art according to the following scheme. However, the method for preparing the compound of Formula 1 is not limited to the following scheme, and may be variously prepared using other known organic synthesis reactions.

In addition, the present invention provides a pharmaceutical composition for the prevention and treatment of cancer diseases comprising the meso-bis (ortho-hydroxyphenyl) bis (para-nitrosi) salen compound or a pharmaceutically acceptable salt thereof. .

In the present invention, the meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen compound induces cell cycle progression inhibition by reducing the amount of cyclin D1 (Cyclin D1) protein to control the growth of cancer cells It is characterized by.

In addition, the meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen compound of the present invention can be synthesized through the synthesis and fractionation of conventional substituents (Herbert O. House, Modern Synthetic Reactions, 2nd Ed). , The Benjamin / Cummings Publishing Co., 1972).

In addition, the compounds of the present invention can be prepared with pharmaceutically acceptable salts and solvates according to methods conventional in the art.

As the pharmaceutically acceptable salt, acid addition salts formed by free acid are useful. Acid addition salts are prepared by conventional methods, for example by dissolving a compound in an excess of aqueous acid solution and precipitating the salt using a water miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. Equal molar amounts of the compound and acid or alcohol (eg, glycol motomethylether) in water can be heated and the mixture can then be evaporated to dryness or the precipitated salts can be suction filtered.

In this case, organic acids and inorganic acids may be used as the free acid, and hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, and the like may be used as the inorganic acid, and methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, Citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid (gluconic acid), galacturonic acid, glutamic acid, glutaric acid (glutaric acid), glucuronic acid (glucuronic acid), aspartic acid, ascorbic acid, carbonic acid, vanic acid, hydroiodic acid may be used.

In addition, bases can be used to make pharmaceutically acceptable metal salts. An alkali metal or alkaline earth metal salt is obtained by, for example, dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and then evaporating and drying the filtrate. In this case, as the metal salt, it is particularly suitable to prepare sodium, potassium or calcium salt, and the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).

Pharmaceutically acceptable salts of the compounds of the invention include salts of acidic or basic groups which may be present in the compounds of the invention, unless otherwise noted. For example, pharmaceutically acceptable salts include sodium, calcium and potassium salts of the hydroxy group, and other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfate, hydrogen sulphate, phosphate, hydrogen phosphate, dihydrogen Phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p-toluenesulfonate (tosylate) salts. It can be prepared through.

Compounds of the present invention obtained by the above method was tested to determine the inhibitory effect of cancer cell proliferation, it can be seen that reducing the amount of cyclin D1 protein expression.

Pharmaceutical compositions comprising a compound of formula 1 according to the invention may further comprise a suitable carrier, excipient or diluent according to conventional methods. Carriers, excipients and diluents that may be included in the compositions of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, Methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The composition according to the invention can be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral formulations, external preparations, suppositories or sterile injectable solutions according to conventional methods.

Specifically, when formulated, it may be formulated using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants and the like that are commonly used. Formulations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient in the compound, such as starch, calcium carbonate, sucrose. ), Lactose, gelatin and the like can be mixed. In addition to the simple excipients, lubricants such as magnesium stearate, talc and the like may also be used. Liquid preparations for oral use include suspensions, solvents, emulsions, and syrups.In addition to the commonly used simple diluents, water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. Can be. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the compounds according to the invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the compound of the present invention may be administered in an amount of 0.0001 to 100 mg / kg, preferably 0.001 to 100 mg / kg, divided once or several times a day. In the compositions of the present invention the compound of formula 1 should be present in an amount of 0.001 to 50% by weight, preferably 0.01 to 50% by weight relative to the total weight of the total composition.

Pharmaceutical dosage forms of the compounds according to the invention can also be used in the form of their pharmaceutically acceptable salts, or can be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.

The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, humans, etc. by various routes, and all modes of administration can be expected, for example, oral, rectal or intravenous, muscle, subcutaneous, It can be administered by intrauterine dural or intracerebroventricular injection.

Hereinafter, the present invention will be described in more detail with reference to Examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

Example. Preparation of meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen compound

Meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen according to the present invention was prepared according to conventional synthetic methods (Chem. Ber., 109 , 1, 1976; Agric. Chem. Biotechnol., 45 , 23, 2002).

Specifically, the acetonitrile solution containing the diamine suspension and the salicylic aldehyde substituted with the nitro functional group were added at a molar ratio of 1: 2, and the mixture was stirred well at room temperature until a clear solution was obtained. After 2 hours, if a yellow precipitate is formed in the reaction mixture, it is filtered and dried to obtain meso-bis (o-hydroxyphenyl) bis (p-nitro). ) salen, 2,2 '-(1E, 1'E)-((1R, 2S) -1,2-bis (2-hydroxyphenyl) ethane-1,2-diyl) bis (azan-1-yl-1 -ylidene) bis (methan-1-yl-1-ylidene) bis (4-nitrophenol)] was obtained in a yield of 86-92%.

¹ H NMR (400 MHz, DMSO) δ 14.79 (br, OH), 10.19 (br, OH), 8.50 (s, H), 8.30 (d, J = 2.9 Hz, H), 8.06 (dd, J = 2.9 , 9.5 Hz, H), 7.31 (dd, J = 1.5, 7.3 Hz, H), 7.13 (td, 1.5, 7.3 Hz, H), 6.84 (m, H), 6.81 (m, H), 6.74 (d , J = 9.5 Hz, H), 5.60 (s, H)

¹³ C NMR (400 MHz, DMSO) d 165.5, 160.2, 155.1, 136.2, 130.4, 129.5, 129.4, 128.7, 123.2, 120.5, 119.3, 115.7, 115.4, 68.3

Experimental Example 1. Confirmation of cyclin D1 expression decrease effect of meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen compound

In the present invention, in order to observe the effect of reducing the CycD1 expression level of the meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen prepared in the above embodiment, the expression level of a specific protein at the cellular stage was measured Western blot assay was performed.

The protein of the cell is 20 mM HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid) buffer solution + 1% Triton X-100 + 10% glycerol + 150 mM NaCl + 10 μg / μl Leupeptin + 1 mM PMSF ( After dissolving in phenylmethylsulfonyl fluoride buffer, the intracellular residues were separated by high-speed centrifugation, and the same amount of protein was separated by electrophoresis on SDS-polyacrylamide gel. The expression level of CycD1 was measured by an enhanced chemiluminescence detection system (Amersham Pharmacia Biotech, Piscataway, NJ) using an antibody against CycD1 protein by transferring to a nitrocellulose filter.

As a result, as shown in Figure 3, the expression of CycD1 significantly reduced after 24 hours in the experimental group treated with meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen of the present invention compared to the control group I could confirm that.

Examples of the pharmaceutical compositions containing the compounds of the present invention will be described, but the present invention is not intended to be limited thereto, but is intended to be described in detail.

Formulation Example 1 Preparation of Powder

20 mg of the compound of the present invention

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

10 mg of compound of the present invention

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3 Preparation of Capsule

10 mg of compound of the present invention

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

10 mg of compound of the present invention

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO ₄ 12H ₂ O 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

20 mg of the compound of the present invention

10 g of isomerized sugar

5 g of mannitol

Purified water quantity

According to the conventional method of preparing a liquid solution, each component is added to the purified water to dissolve it, and lemon flavor is added, the above components are mixed, and then, purified water is added to adjust the total amount to 100 ml, and then sterilized by filling into a brown bottle. do.

1 shows hydrogen nuclear magnetic resonance spectroscopy spectra of meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen according to the present invention.

2 shows carbon nuclear magnetic resonance spectroscopy spectra of meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen according to the present invention.

3 is a result of Western blot confirming the effect of reducing the amount of expression for cyclin D1 (CycD1) protein of meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen according to the present invention.

Claims (4)

Meso-Bis (o-hydroxyphenyl) meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen of formula 1 to suppress abnormal cell division and proliferation by reducing the expression level of Cyclin D1 (Cyclin D1) protein bis (p-nitro) salen, 2,2 '-(1E, 1'E)-((1R, 2S) -1,2-bis (2-hydroxyphenyl) ethane-1,2-diyl) bis (azan -1-yl-1-ylidene) bis (methan-1-yl-1-ylidene) bis (4-nitrophenol)] compound. [Formula 1]

A pharmaceutical composition for preventing and treating cancer diseases comprising the meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen compound of claim 1 or a pharmaceutically acceptable salt thereof. 3. The method of claim 2, The meso-bis (ortho-hydroxyphenyl) bis (para-nitro) salen compound is a pharmaceutical composition characterized in that by inhibiting the cell cycle progression by reducing the amount of cyclin D1 protein expression to control the proliferation of cancer cells. 3. The method of claim 2, The cancer diseases include gastric cancer, colon cancer, breast cancer, lung cancer, non-small cell lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, anal muscle cancer, fallopian tube death, endometrium Carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penis cancer, prostate cancer, chronic or Acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brain stem glioma, or pituitary adenoma Pharmaceutical composition.

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