KR20190124951A - Compositions for preventing or treating kidney cancer comprising PFI-3 - Google Patents

Abstract

The present invention relates to a composition for preventing or treating renal cancer, comprising PFI-3. Specifically, the present invention relates to: a pharmaceutical composition for treating renal cancer; a health functional food composition comprising PFI-3 or a pharmaceutically acceptable salt thereof; and a method for treating renal cancer comprising a step of administering the pharmaceutical composition to a subject. PFI-3 of the present invention not only exhibits an excellent killing effect on renal cancer cells, but also can sustain the killing effect for a long time, thereby being able to be usefully used for preventing, alleviating, or treating renal cancer.

Description

Compositions for preventing or treating kidney cancer comprising PFI-3

The present invention relates to a composition for the prevention or treatment of kidney cancer, including PFI-3, specifically, comprising a pharmaceutical composition for the treatment of kidney cancer, including PFI-3 or a pharmaceutically acceptable salt thereof; Health functional food composition; And administering the pharmaceutical composition to a subject.

Kidney cancer is a malignant tumor that occurs in the parenchyma of the kidney, which consists of the medulla and cortex where urine-producing cells are gathered. When the size of the tumor is small, there are few symptoms, and the tumor grows to a certain extent until the organ is pushed out until the symptoms appear. Therefore, the diagnosis is often delayed, and when the first diagnosis is made, about 30% of the patients are already metastasized to the lung, brain, liver, and bone. The most common symptom is hematuria, but this only occurs in 60% of patients. Rather, depending on the metastases, symptoms such as difficulty breathing, coughing, and headache appear, which causes kidney cancer to be diagnosed in 30% of patients.

In general, kidney cancer cells are known to be very susceptible to radiation or chemotherapy. Thus, the treatment of kidney cancer is mainly performed by surgical resection. However, in this case, there is a problem in that not only tumors but also normal tissues around them are extensively excised, and studies on the development of drugs targeting kidney cancer are being actively conducted. For example, a pharmaceutical composition for the prevention or treatment of kidney cancer comprising a high dose Kang extract (Korean Patent Publication No. 10-2018-0026675), a composition for the treatment or prevention of liver cancer or kidney cancer comprising sorafenib and gold silver extract ( Korean Patent Publication No. 10-1833036), a composition for treating renal cancer comprising apilimod (Korean Patent Publication No. 10-2017-0098812) and the like has been developed. However, there is still a need to develop more effective kidney cancer therapeutic drugs that can overcome the low drug sensitivity of kidney cancer.

Against this background, the present inventors have made intensive studies to develop drugs that can effectively treat kidney cancer. As a result, PFI-3 compounds have shown a high survival rate reduction effect against various kidney cancer cell lines. The present invention was completed by confirming that it is exerted within a short time and maintained for a long time.

One object of the present invention is to provide a pharmaceutical composition for treating kidney cancer, comprising PFI-3 or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a method for treating kidney cancer, comprising administering the pharmaceutical composition to a subject other than a human.

Yet another object of the present invention is to provide a dietary supplement composition for improving kidney cancer, comprising PFI-3 or a physiologically acceptable salt thereof.

In order to achieve the above object, one embodiment of the present invention provides a pharmaceutical composition for treating kidney cancer, comprising PFI-3 or a pharmaceutically acceptable salt thereof.

PFI-3 compound provided by the present invention alone has an anticancer effect against a variety of renal cell carcinoma, there is an advantage that can maintain the effect for a long time of about 10 days or more with a single treatment.

As used herein, the term "PFI-3" refers to a compound represented by the following Chemical Formula 1.

[Formula 1]

The molecular weight of the PFI-3 is 321.37, the chemical formula is C ₁₉ H ₁₉ N ₃ O ₂ , the chemical name is (2E) -1- (2-hydroxyphenyl) -3-[(1R, 4R) -5- (pyridine- 2-yl) -2,5-diazabicyclo [2.2.1] heptann-2-yl] prop-2-ene-1-one, CAS number known to 1819363-80-8. Currently, studies on the pharmacological activity of PFI-3 are insignificant, and its anticancer effect was first identified by the present inventors.

The method for obtaining PFI-3 is not particularly limited, and chemically synthesized using a method known in the art, or a commercially available material may be used.

In addition, the PFI-3 may exist in solvated or unsolvated form, and may exist in crystalline or amorphous form, and all such physical forms are included in the scope of the present invention.

In the present invention, the PFI-3 may be to inhibit the survival of renal cell carcinoma 786-O, UMRC-6 or Caki-2 cells.

There are more than 40 subtypes of renal cell carcinoma, which are classified into clear cell carcinoma, papillary carcinoma, and chromophobe carcinoma. Among them, clear cell carcinoma accounts for 70-80% of all renal cell carcinomas and has the highest incidence. Clear cell carcinoma is also classified into H1H2 type, H2 type and the like according to its genetic characteristics, H1H2 type expresses both HIF1αα and HIF2αα protein, H2 type is known to express only HIF2αα protein and HIF1αα protein does not function.

Specifically, "786-O" and "UMRC-6" is a H2 type renal cell carcinoma, "Caki-2" is a H1H2 type renal cell carcinoma, the PFI-3 is regardless of the type of renal cell carcinoma Can exhibit anticancer efficacy.

On the other hand, in the present specification, the renal cell carcinoma may be used favorably in the same sense as kidney cancer cells.

In one embodiment of the present invention, PFI-3 shows the effect of reducing the survival rate of various kidney cancer cells, such as 786-O, UMRC-6, Caki-2, such a cancer cell death effect is a short period of about 2 days In addition, it was confirmed that only one treatment at the beginning of the culture can maintain the effect for a long period of about 10 days (FIGS. 1 to 5).

This suggests that the pharmaceutical composition of the present invention containing PFI-3 may be usefully used for the prevention, treatment, and the like of kidney cancer.

As used herein, the term “treatment” means any action in which symptoms of kidney cancer are improved or beneficially altered by administration of a composition comprising the PFI-3 or a pharmaceutically acceptable salt thereof.

As used herein, the term “pharmaceutically acceptable salts” refers to salts that can be used pharmaceutically, even among salts in which cations and anions are bound by electrostatic attraction. Salts, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, and the like. For example, the metal salt may be an alkali metal salt (sodium salt, potassium salt, etc.), alkaline earth metal salt (calcium salt, magnesium salt, barium salt, etc.), aluminum salt, or the like; Salts with organic bases include triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, N, N-dibenzylethylenediamine Salts and the like; Salts with inorganic acids can be salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, and the like; Salts with organic acids can be salts with formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid and the like; Salts with basic amino acids can be salts with arginine, lysine, ornithine and the like; Salts with acidic amino acids can be salts with aspartic acid, glutamic acid and the like.

The pharmaceutical composition of the present invention may include PFI-3 or a pharmaceutically acceptable salt thereof in an amount of 0.01 to 80%, specifically 0.01 to 70%, and more specifically 0.01 to 60% by weight, based on the total weight of the composition. As long as it shows a therapeutic effect of cancer, it is not limited thereto.

In addition, the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier, excipient or diluent commonly used in the manufacture of the pharmaceutical composition, the carrier may comprise a non-naturally occuring carrier (carrier) have. Specific examples of the carriers, excipients and diluents 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 or mineral oil may be used, but is not limited thereto.

In addition, the pharmaceutical composition may be a tablet, a pill, a powder, a granule, a capsule, a suspension, a solution, an emulsion, a syrup, a sterile aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a lyophilizer and a suppository, respectively, according to a conventional method. It may have any one formulation selected from the group consisting of, and may be various oral or parenteral formulations. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Tablets, pills, powders, granules, capsules and the like may be used for the solid preparation for oral administration, and the solid preparation may be at least one excipient such as starch, calcium carbonate, sucrose or lactose, Gelatin and the like can be used. In addition to the simple excipients, lubricants such as magnesium stearate, talc and the like can be used. As a liquid preparation for oral administration, suspending agents, liquid solutions, emulsions, syrups, etc. may be used.In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be used. Can be used. For parenteral administration, sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations or suppositories may be used. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like may be used, but are not limited thereto.

Another aspect provides the method of treating kidney cancer, comprising administering the pharmaceutical composition to a subject other than a human.

At this time, the description of the "treatment" is as described above.

As used herein, the term "individual" means all animals, such as rats, mice, and livestock, including humans, who may or may have kidney cancer. As a specific example, it may be a mammal including a human.

As used herein, the term "administration" means introducing the pharmaceutical composition to an individual in a suitable manner.

The pharmaceutical composition of the present invention may be administered in a pharmaceutically effective amount.

The term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and an effective dose level refers to the type and severity of the subject, severity, age, sex, activity of the drug, Sensitivity to drug, time of administration, route of administration and rate of release, duration of treatment, factors including concurrent use of drugs, and other factors well known in the medical arts.

The pharmaceutical composition may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. In addition, single or multiple administrations can be made. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect in a minimum amount without side effects, which can be easily determined by those skilled in the art.

In addition, the pharmaceutical composition may be administered orally or parenterally (eg, applied intravenously, subcutaneously, intraperitoneally, or topically) according to a desired method, and the dosage is based on the condition and weight of the patient, and the degree of disease. Depending on the drug form, route of administration, and time, it may be appropriately selected by those skilled in the art. As a specific example, the pharmaceutical composition may generally be administered in an amount of 0.001 to 1000 mg / kg, more specifically 0.05 to 200 mg / kg, most specifically 0.1 to 100 mg / kg once a day or several times. Preferred dosages may be appropriately selected by those skilled in the art depending on the condition and weight of the individual, the severity of the disease, the form of the drug, the route of administration and the duration.

Another aspect provides a dietary supplement composition for improving kidney cancer, comprising PFI-3 or a physiologically acceptable salt thereof.

At this time, the definition of "PFI-3" is as described above.

As used herein, the term “physiologically acceptable salts” is a physiologically acceptable and when administered to an organism, the compound to be administered does not normally cause an allergic or similar reaction, such as gastrointestinal disorders, dizziness, or the like. It means that it is commonly used to exhibit the effect.

As used herein, the term "improvement" refers to any action that at least reduces the parameters associated with the condition to be treated with the administration of a composition comprising PFI-3, such as the extent of symptoms.

PFI-3 according to the present invention shows an excellent survival rate reduction effect on the kidney cancer cells, can be included in the dietary supplement composition for the purpose of preventing or improving kidney cancer, the dietary supplement composition is to be taken daily Because of this, high effects can be expected for the prevention or improvement of kidney cancer.

As used herein, the term "health functional food" refers to a food prepared and processed using raw materials or ingredients having functional properties useful for the human body according to Act No. 6767 of the Health Functional Food Act, and "functionality" refers to the structure of the human body. And it means to obtain a useful effect for health use, such as regulating nutrients or physiological action for function. On the other hand, health food refers to foods that have active health maintenance or promotion effect compared to general foods, and health supplement food means foods for the purpose of health supplement, in some cases, the term functional health food, health food, health supplement food Can be used interchangeably.

PFI-3 of the present invention may be added as it is or used with other foods or food ingredients, and may be suitably used according to conventional methods.

The health functional food composition of the present invention can be prepared by a method commonly used in the art, and the preparation can be prepared by adding raw materials and ingredients commonly added in the art. Specifically, the nutraceutical composition may further include a physiologically acceptable carrier, the type of carrier is not particularly limited and may be used as long as it is a carrier commonly used in the art.

In addition, the health functional food composition is a food such as preservatives, fungicides, antioxidants, colorants, coloring agents, bleaches, seasonings, sweeteners, flavorings, swelling agents, reinforcing agents, emulsifiers, thickeners, coatings, gum herbicides, foam inhibitors, solvents, improvers May include additives. The additive may be selected according to the type of food and used in an appropriate amount.

In addition, the formulation of the health functional food can be prepared without limitation as long as the formulation is recognized as food. The health functional food composition of the present invention can be prepared in a variety of dosage forms, unlike the general medicine because it has the advantage that there is no side effect, such as may occur when taking a long-term use of the food as a raw material and excellent portability, The food of the present invention can be taken as an adjuvant for enhancing the effect of preventing or improving kidney cancer.

PFI-3 of the present invention may be included in various weight percent in the dietary supplement composition if it can exhibit an improvement effect of kidney cancer. Specifically, it may be included as 0.00001 to 100% by weight or 0.01 to 80% by weight relative to the total weight of the dietary supplement composition, but is not limited thereto. When ingested for a long time may contain a content below the above range, it may be used in an amount above the above range.

In one embodiment of the present invention, PFI-3 has the effect of reducing the survival rate of various kidney cancer cells, the cancer cell killing effect is shown in a short period of about 2 days, and also the effect of a single treatment at the beginning of the culture It was confirmed that can be maintained for a long period of about 10 days (Figs. 1 to 5).

This suggests that the health functional food composition of the present invention containing PFI-3 can be usefully used for the prevention, treatment, and the like of kidney cancer.

PFI-3 of the present invention not only exhibits an excellent killing effect on kidney cancer cells but can also maintain the effect for a long time, and thus can be usefully used for preventing, improving, and treating kidney cancer.

1 is a diagram showing the anticancer effect of PFI-3 against 786-O kidney cancer cells, A is a schematic diagram showing the PFI-3 treatment scheme, B is Crystal Violet staining showing the colony forming assay results Image, and C are graphs quantifying the results according to staining. PFI-3 was treated once on the second day of culture.
2 is a diagram showing the anticancer effect of PFI-3 against 786-O kidney cancer cells, A is a schematic showing the PFI-3 treatment scheme, B is a crystal violet staining image showing the colony formation assay results, and C is It is a graph quantifying the results according to staining. PFI-3 was treated once each on days 2, 5 and 8 of culture.
3 is a graph showing the anticancer effect of PFI-3 against 786-O renal cancer cells, a graph showing the results of the MTS assay.
4 is a graph showing the anticancer effect of PFI-3 on UMRC-6 kidney cancer cells, a graph showing the MTS assay results.
5 is a graph showing the anticancer effect of CFI-2 renal cancer cells of PFI-3, a graph showing the results of the MTS assay.

Hereinafter, the present invention will be described in detail by way of 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  One: Renal cell carcinoma  For 786-O cells PFI -3 anticancer effect confirmed

In order to confirm the effect of PFI-3 on the growth of renal cancer cells, 786-O cells were treated with PFI-3 to investigate the survival rate of cancer cells. First, the degree of death of renal cancer cells by PFI-3 single treatment was confirmed.

Specifically, 786-O cells were treated with PFI-3 at a concentration of 30 to 90 μM once during the culture period, and colony formation assay was performed to confirm long-term cytotoxicity. The viability of the cells was analyzed. After completion of 10 days of incubation the colonies were rinsed with 1X PBS and visualized using 5% crystal violet contained in 10% methanol.

As a result, as can be seen in Figure 1, as the concentration of PFI-3 increases the survival rate of 786-O cells decreases to about 40% was reduced by 90 μM treatment, in particular, about 10 days PFI-3 It was confirmed that the effect was maintained even after only one treatment for the above long period.

In addition, it was intended to determine whether the same anti-cancer effect even when treated repeatedly PFI-3.

Specifically, PFI-3 was treated in 786-O cells once every three days for 10 days of culture at a concentration of 30 to 90 μM, and colony formation assays were performed to analyze the viability of 786-O cells.

As a result, as can be seen in Figure 2, as the concentration of PFI-3 increases the survival rate of 786-O cells decreases by about 15% when 90 μM treatment, confirming that the cell killing effect is increased compared to the single treatment It was.

The results indicate that PFI-3 exhibits an excellent anticancer effect against 786-O kidney cancer cells, and can be maintained for about 10 days or longer with a single treatment.

Example  2: various In renal cell carcinoma  About PFI -3 anticancer effect confirmed

Example 1 confirmed the anticancer effect on 786-O cells, in addition, to determine whether PFI-3 has the same anti-effect against a variety of renal cell carcinoma.

This used 786-O, UMRC-6, Caki-2 cells. 786-O, UMRC-6 is H2 type renal cell carcinoma, Caki-2 is H1H2 type renal cell carcinoma, BAF180 is deleted, PFI-3 in all of the various renal cell carcinoma having different characteristics It was confirmed to have an anticancer effect.

Specifically, PFI-3 was treated in 786-O, UMRC-6, and Caki-2 cells at a concentration of 30 to 120 μM, and after completion of 2 days of culture, MTS assay was performed to analyze the survival rate of cancer cells. . After completion of the two-day incubation MTS assay was performed according to the existing protocol (CellTilter 96 aqueous One Solution Cell Proliferation assay, Promega). After treatment with the MTS solution and incubated for 1 hour, the absorbance was measured at 490 nm using a spectraMax i3X plate reader (I3X-SC-ACAD, Molecular Devices).

As a result, as shown in Figures 3 to 5, PFI-3 shows an excellent cell killing effect on all 786-O, UMRC-6, Caki-2 cells, the higher the concentration of PFI-3, the higher the survival rate of the cells This decrease was confirmed.

From the above results, it was found that PFI-3 has excellent anticancer effects on UMRC-6 and Caki-2 cells as well as 786-O, and can have a significant killing effect in a short period of about 2 days.

In addition, the synthesis of the contents of Examples 1 and 2, PFI-3 shows an excellent anti-cancer effect on kidney cancer cells, the effect was found to be exerted in a short time and lasts a long time.

From the above description, those skilled in the art will appreciate that the present invention can be implemented in other specific forms without changing the technical spirit or essential features. In this regard, the embodiments described above are to be understood in all respects as illustrative and not restrictive. The scope of the present invention should be construed that all changes or modifications derived from the meaning and scope of the following claims and equivalent concepts rather than the detailed description are included in the scope of the present invention.

Claims (5)

A pharmaceutical composition for treating kidney cancer, comprising PFI-3 or a pharmaceutically acceptable salt thereof.
The pharmaceutical composition of claim 1, wherein the PFI-3 is represented by the following Chemical Formula 1.
[Formula 1]

The pharmaceutical composition of claim 1, wherein the PFI-3 inhibits the survival of renal cell carcinoma 786-O, UMRC-6 or Caki-2 cells.
A method of treating kidney cancer, comprising administering the pharmaceutical composition of any one of claims 1 to 3 to a subject other than a human.
A dietary supplement for the improvement of kidney cancer, comprising PFI-3 or a physiologically acceptable salt thereof.

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