KR20100023676A - Use in anti-cancer agent of diphenyl phthalate - Google Patents

Abstract

PURPOSE: A diphenyl phthalate compound is provided to ensure an effect of inhibiting the growth of cells by activating the expression of p21 as a cell cycle proceeding inhibitor, to inhibit the proliferation of cancer cells, and to induce apoptosis. CONSTITUTION: A diphenyl phthalate compound has a structure represented by chemical formula 1. A pharmaceutical composition for the prevention and treatment of cancer disease comprises the diphenyl phthalate compound and pharmaceutically acceptable salts thereof. The cancer disease is cancer caused by abnormality of p21 genes. The pharmaceutically acceptable salts are acid addition salts formed by free acids.

Description

Use of anti-cancer agent of diphenyl phthalate

The present invention relates to the use of a diphenyl phthalate compound as an anticancer agent, and more particularly, the diphenyl phthalate compound represented by Chemical Formula 1 below to inhibit the proliferation of cancer cells and induce cell death by activating p21 expression. It relates to a pharmaceutical composition for the prevention and treatment of cancer diseases, including the compound or a pharmaceutically acceptable salt thereof.

p21 is a Cyclin Dependent Kinase Inhibitor 1A, which is a cyclin / cyclin / cyclin-dependent kinase 2 (Cyclin-CDK4) or a cyclin-CDK4 (Cyclin-CDK4) It is a protein that inhibits the activity of CDK complexes.

Various cyclins form complexes with CDKs and are involved in cell growth and differentiation. The cyclins are divided into two phases, the inter phase and the mitotic phase. G (1) -phase to synthesize various proteins, S-phase to synthesize DNA, growth for mitosis, G (2) -phase to prepare, and mitosis-phase to fix cell differentiation Etc. It consists of four courses.

In other words, various cyclin-CDK complexes are involved in this process to regulate cell cycle progression.

At each stage of the cell cycle process, there are check points to check that each stage is working properly, such as checking for DNA damage before proceeding. p21 inhibits cell cycle progression activity, in particular by binding to cyclin-CDK complexes involved in the checkpoint when progressing from G (1) -phase to S-phase. This function interacts with transcription factors that promote cell cycle progression, thereby inhibiting the phosphorylation of pRb (Retinoblastoma protein), which inhibits cell cycle progression, thereby maintaining an active state. It prevents the progression from -phase to S-phase and, consequently, prevents cell growth and differentiation.

In addition, p21 directly prevents DNA replication by interacting independently with Proliferating Cell Nuclear Antigen (PCNA), a protein that acts as a progressive factor for DNA polymerase delta.

Cell cycle suppression stops the cell's differentiation and restores damage, thereby allowing the cell's differentiation to continue normally.However, unlike normal cells, cancer cells are cells that do not properly regulate growth-differentiation and self-killing functions. Therefore, it is possible to suppress the proliferation of cancer cells through cell cycle inhibition.

Therefore, a large number of studies have shown that the decrease in the expression of p21, which indirectly plays a role in cell cycle suppression, is closely related to the progression or metastasis of cancer. Increasing the expression of p21 as a cancer inhibitor by inhibiting the proliferation of cells is very meaningful because it is to search for substances having a structure different from existing anticancer substances based on natural products.

Accordingly, the present inventors have diligently tried to find a substance capable of inhibiting the proliferation of cancer cells by overexpressing p21, a cell cycle inhibitor, and inhibiting cell growth and differentiation. As a result, diphenyl phthalate activates p21 expression. It confirmed and completed this invention.

After all, the present invention has a main purpose to provide a diphenyl phthalate compound having an activity of increasing expression for the p21 gene and the use of the compound as an anticancer agent.

In order to achieve the above object, the present invention provides a diphenyl phthalate compound of Formula 1 to inhibit the proliferation of cancer cells and induce cell death by activating p21 expression.

[Formula 1]

The present invention also provides a use of the diphenylphthalate compound as an anticancer agent.

Specifically, the present invention provides a pharmaceutical composition for the prevention and treatment of cancer diseases comprising the diphenyl phthalate or a pharmaceutically acceptable salt thereof.

The cancer disease includes a general cancer disease caused by abnormalities of the p21 gene, 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 eye melanoma , Uterine cancer, ovarian cancer, rectal cancer, anal muscle cancer, fallopian tubes, 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, urethral cancer, penile 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, 1 One or more diseases selected from primary CNS lymphoma, spinal cord tumor, brain stem glioma, pituitary adenoma, and the like.

Diphenyl phthalate of the present invention has the effect of inhibiting cell growth by activating p21 expression, a cell cycle progression inhibitor, thereby inhibiting the proliferation of cancer cells and inducing apoptosis, thereby preventing and treating cancer. It can be usefully used.

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

The present invention provides a diphenyl phthalate compound of Chemical Formula 1, which inhibits the proliferation of cancer cells and induces apoptosis by activating the expression of p21.

[Formula 1]

In the present invention, the diphenyl phthalate of Chemical 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, including the diphenylphthalate compound or a pharmaceutically acceptable salt thereof.

In the present invention, the diphenylphthalate compound is characterized by controlling the proliferation of cancer cells by activating p21 expression to inhibit cell cycle and induce apoptosis.

In addition, the diphenylphthalate compound of the present invention can also 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, glucuronic acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanic acid, hydroiodic acid, and the like.

Bases can also 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.

The compound of the present invention obtained by the above method was tested to determine the effect of inhibiting the proliferation of cancer cells and inducing apoptosis, can confirm the expression activity of p21.

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 compositions 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. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized 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 Diphenyl Phthalate

In a 100 ml round bottom flask connected to a Dean-Stark trap and condenser, 4.9 g (0.03 mol) of phthalic anhydride, 5.6 g (0.06 mmol) of phenol, 3.3 mg of paratoluenesulfonic acid (p-TsOH) , 0.017 mmol) was added dropwise and 20 ml of toluene was added dropwise and stirred under reflux for 3 hours.

After cooling to room temperature, triethylamine was added, the mixture was distilled under reduced pressure to remove volatiles, and the residue was purified by silica gel column chromatography to obtain 1.9 g (6.0 mmol, 20% yield) of diphenylphthalate.

¹ H NMR (400 MHz, MeOD) δ 7.96-7.98 (m, 2H), 7.65-7.69 (m, 2H), 7.36-7.40 (m, 4H), 7.21-7.26 (m, 6H)

¹³ C NMR (100 MHz, MeOD) d 166.3, 151.2, 132.22, 132.19, 130.0, 126.6, 122.0.

Experimental Example 1. Confirmation of increased expression of p21

In the present invention, in order to observe whether or not the expression of p21 of the diphenylphthalate prepared in Example, Western blot assay to measure the expression level of a specific protein using the antibody of the protein in the cell stage 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 p21 was measured with an enhanced chemiluminescence detection system (Amersham Pharmacia Biotech, Piscataway, NJ) using an antibody against p21 transferred to a nitrocellulose filter.

As a result, as shown in Figure 2, the expression of p21 protein in the experimental group treated with the diphenyl phthalate of the present invention compared with the control group time-dependently increased, especially after 24 hours increased 12 times more than 12 hours Confirmed.

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

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.

Figure 1 shows the hydrogen nuclear magnetic resonance spectroscopy spectrum of diphenyl phthalate according to the present invention.

Figure 2 shows the carbon nuclear magnetic resonance spectroscopy spectrum of diphenylphthalate according to the present invention.

Figure 3 is a Western blot confirming the effect of the expression activity on the p21 gene of diphenylphthalate according to the present invention.

Claims (5)

A diphenyl phthalate compound represented by the following Chemical Formula 1, which inhibits proliferation of cancer cells and induces apoptosis by activating p21 expression. [Formula 1]

A pharmaceutical composition for the prevention and treatment of cancer diseases comprising the diphenylphthalate compound of claim 1 or a pharmaceutically acceptable salt thereof. 3. The method of claim 2, The diphenyl phthalate compound is a pharmaceutical composition, characterized in that to control the proliferation of cancer cells by activating p21 expression inhibits the cell cycle, and induces apoptosis. 3. The method of claim 2, The cancer disease is a pharmaceutical composition, characterized in that the cancer caused by abnormalities of the p21 gene. The method according to claim 2 or 4, 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, renal 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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