KR101771364B1 - Pharmaceutical composition for the prevention or treatment of lung cancer comprising sanggenol Q - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating lung cancer comprising sanggenol Q. The pharmaceutical composition for preventing or treating lung cancer comprising sanggenol Q according to the present invention is effective in the prevention or treatment of lung cancer by activating caspase 3 and caspase 9, promoting cleavage of poly-ADP ribose polymerase (PARP), and inhibiting Bcl-2, XIAP, and Bad in lung cancer cells.

Description

[0001] The present invention relates to a pharmaceutical composition for prevention or treatment of lung cancer,

The present invention relates to a pharmaceutical composition for preventing or treating lung cancer.

The most common cancer in Korea is gastric cancer, colorectal cancer, lung cancer, thyroid cancer, and liver cancer. Based on national cancer incidence statistics, the risk of developing cancer is increasing, as one in three men and one in four women are diagnosed with a lifetime cancer risk. Especially, in the case of lung cancer, it is expected that the number of lung cancer patients will be increased in view of the deterioration of air pollution caused by industrialization, and the increase of smokers, especially the increase of smoking among adolescents and women. In the United States, lung cancer is the most common cause of cancer-related deaths for both men and women.

The major forms of lung cancer are small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC). Small cell lung cancer (SCLC) is a rapidly growing type of lung cancer. It spreads much faster than non-small cell lung cancer. There are three different types of small cell lung cancer: small cell carcinoma (ochocardiocarcinoma), mixed small cell / large cell carcinoma and multiple small cell carcinoma. Most small cell lung cancer is ovarian cell type. Non-small cell lung cancer (NSCLC) is the most common type of lung cancer. There are three types of NSCLC: adenocarcinoma, squamous cell acinoma, and large-cell carcinoma. Adenocarcinoma occurs mainly in the lung peripheral region, occurs well in women or nonsmokers, and is often metastasized even when the size is small. Recently, the incidence of adenocarcinoma is increasing. Next, squamous cell carcinoma is found mainly in the central lung, and it is known to grow up to the lumen of the bronchial tree, blocking the airway, is common to men, and is closely related to smoking. Large cell carcinomas occur predominantly in the lungs near the lungs (peripheral lungs), about half in large bronchi, accounting for 4 to 10% of all lung cancers, with generally large cell size, It is known that the prognosis is worse than other non - small cell lung cancer. Conventional treatments for lung cancer include palliative care, surgery, chemotherapy, and radiation therapy.

When lung cancer is diagnosed at an early stage, it can be recovered through drug therapy or radiation therapy. However, if the disease progresses to a certain level, the lesion is removed through surgery, and medication and radiation therapy Treatment is performed. However, since the drugs approved to be used for the treatment of lung cancer are limited to a few drugs including Iressa and the like, it is difficult to perform appropriate treatment for patients, and these drugs have disadvantages in that they exhibit negligible levels of side effects. The subsequent success rate of treatment is poor. In order to overcome these disadvantages, attempts have been made to develop therapeutic agents derived from complex natural substances that can reduce side effects. However, the fundamental drawback of these therapeutic agents derived from natural products has not been solved.

Therefore, it is necessary to develop a formulation that can treat lung cancer more safely and effectively.

The inventors of the present invention confirmed that sanggenol Q obtained from the extract of Mulberry bark extract had a remarkable effect on the prevention and treatment of lung cancer in order to develop a therapeutic agent capable of treating safe and effective lung cancer, Thereby completing the invention.

Giaccia A. et al. Nat Rev Drug Disc 2; 803-811, 2003

The present invention is to provide a pharmaceutical composition for preventing or treating lung cancer comprising sanggenol Q.

The present invention also provides a health functional food for preventing or ameliorating lung cancer comprising sanggenol Q.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating lung cancer comprising sanggenol Q.

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

Whenever a component is referred to as "comprising ", it is to be understood that the component may include other components as well, without departing from the scope of the present invention.

As used herein, the term "prevention" means any action that inhibits or delays lung cancer by administration of the pharmaceutical composition. The term "treatment" as used herein means all the actions of improving or alleviating symptoms of lung cancer by the administration of the pharmaceutical composition.

In the present specification, lung cancer means lung cancer, and may be small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC), and the small cell lung cancer and non-small cell lung cancer are as described above.

The sanggenol Q of the present invention means a compound represented by the following general formula (1).

[Chemical Formula 1]

The Sangenol Q can be isolated from Morus alba L. or purchased commercially.

In the present specification, the term "bark bark" refers to a medicinal substance made of the root bark of mulberry or an inbred plant. The bark bark treats seawater and asthma caused by waste heat and has a diuretic effect. It is used for acute pyelonephritis, weakness edema, hypotensive effect, and also for nosebleed and blood. It is also used for epidemic hepatitis. The pharmacological action was reported to be Jinhae, diuretic, hypotension, sedation, analgesia, fever, epidermal and antimicrobial effects. Morus alba L. contains main ingredients such as water berlin, water verrucroven, cyclo water berlin and coumarin derivatives such as scopoletin, umbelliferone, triterpenoid aminoline and betulinic acid. , Vinegar action, antibacterial action, anti-inflammatory action, antioxidant effect and is known as a good medicine.

Morus alba L. and Morus alba L. Remove the cork layer from the root of the root of the plant. In the winter, wash the roots cleanly, and when it is fresh, peel off the layer of cork, cut it vertically and cut it with a wooden hammer, It is only taken. The taste is sweet and the quality is cold. Alcoholic beverages have been used in oriental medicine for the treatment of diabetes, diuretics, pills and diarrhea (Giaccia A. et al. Nat Rev Drug Disc 2; 803-811, 2003). These compounds include several compounds such as prenylated flavonoid, benzofuran, and phenol compounds. These compounds inhibit COX-1, COX-2, and nitric oxide (NO) production Various biological activities have been reported. The major physiologically active ingredients include Umbelliferone, Mulberrochromene, Cyclomulberrin, Cyclomulberrochromene, etc., and it acts with acetylcholine This similar anti-inflammatory component is contained.

Sangenol Q in the present invention can be obtained through extraction and separation of bark extract, and extraction and separation of the bark extract can be performed by methods used in the art, but are not limited thereto.

Herein, the extraction refers to the separation of an active ingredient from a natural product, and an extract can be obtained by an extraction process using water, an organic solvent or a mixed solvent thereof, and the extract can be obtained by using an extract, a dry powder thereof, It can include all forms that have been formalized.

For example, the extract solution of the plant extract of the present invention can be extracted with water or an organic solvent. Examples of the organic solvent include lower alcohols, acetone, chloroform, methylene chloride, ether, ethyl acetate, hexane and the like. As the lower alcohol, methanol, ethanol, propanol and butanol can be mentioned, and ethanol is most preferable.

The pharmaceutical composition of the present invention may include, but is not limited to, 0.1 to 90% by weight of sanggenol Q based on the total weight of the pharmaceutical composition for preventing or treating lung cancer.

In the pharmaceutical composition for preventing or treating lung cancer comprising sanggenol Q of the present invention, prevention or treatment of lung cancer is caused by promotion of lung cancer cell death.

Specifically, a pharmaceutical composition comprising sanggenol Q of the present invention activates a pro-apoptotic protein. For example, the expression of proteins of PARP and cleaved caspase 3 is increased while that of procaspase 8, procaspase 9, XIAP, BAD and Bcl-2 is increased by expression of sanggenol Q . In addition, the pharmaceutical composition comprising Sangenol Q of the present invention inhibits the expression of Shh and SUFU, genes associated with hedgehog signals involved in cancer metastasis, while PTCH-1, PTCH-2 and 0.0 > GLI < / RTI >

Accordingly, the pharmaceutical composition for preventing or treating lung cancer including sanggenol Q of the present invention can exert an excellent effect for prevention and / or treatment of lung cancer by promoting apoptosis of lung cancer.

The pharmaceutical composition for preventing or treating lung cancer comprising sanggenol Q of the present invention is a pharmaceutical composition.

The pharmaceutical composition of the present invention does not increase the drug efficacy but may further include components that are commonly used in pharmaceutical compositions and can improve odor, taste, and visual appearance. In addition, the pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable additive. Pharmaceutically acceptable additives include, for example, starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, mannitol, gum, arabic gum, pregelatinized starch, cornstarch, powdered cellulose, But are not limited to, hydroxypropylcellulose, opaques, sodium starch glycolate, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, white sugar, dextrose, sorbitol and talc. In addition, the pharmaceutical composition may contain a substance that has a prophylactic or therapeutic activity against lung cancer used alone or in a conventional manner.

The pharmaceutical composition of the present invention comprises a pharmaceutically acceptable carrier and may be formulated for oral or parenteral administration for human or veterinary use. When the pharmaceutical composition of the present invention is formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants may be used. Solid form preparations for oral administration include tablets, pills, powders, granules and capsules, which may be prepared by mixing the pharmaceutical composition comprising the compound of the present invention with at least one excipient such as starch, calcium carbonate Calcium carbonate, Sucrose or Lactose, and Gelatin. In addition to simple excipients, lubricants such as magnesium, styrene, and talc may be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions and syrups, and various excipients such as wetting agents, sweetening agents, fragrances and preservatives in addition to commonly used simple diluents such as water and liquid paraffin . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. Examples of non-aqueous solvents and suspensions include vegetable oils such as propylene glycol, polyethylene glycol and olive oil, injectable esters such as ethyl oleate, and the like. Examples of suppository bases include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like.

The pharmaceutical composition of the present invention may be administered orally or parenterally in accordance with the desired method, and may be administered orally, parenterally or intraperitoneally, rectally, subcutaneously, intravenously, intramuscularly, It is preferable to select the injection method.

The pharmaceutical composition of the present invention can be administered to an individual to prevent or treat lung cancer. The term "individual" as used in the present invention refers to a disease caused by lung cancer or a direct or indirect cause thereof, and includes a human having a disease in which symptoms can be alleviated by administering the pharmaceutical composition of the present invention , Sheep, pigs, goats, dogs and the like, preferably humans.

As used herein, the term "administering" means introducing a pharmaceutical composition of the invention into a subject by any suitable method. The route of administration may be oral or parenteral administration via any conventional route so long as it can reach the target tissue. In addition, the pharmaceutical composition of the present invention may be administered by any device that allows it to migrate to a target cell.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The term, pharmaceutically effective amount, as used herein, means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and an effective dosage level will vary with the weight, sex, age, , The activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and rate of release, the duration of the treatment, factors including co-administered drugs, and other factors well known in the medical arts. The pharmaceutical composition of the present invention 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. May be administered singly or multiply. It is important to take into account all of the factors mentioned above and to administer the amount in which the maximum effect can be obtained in a minimal amount without adverse effect, and it can be easily determined by those skilled in the art. Generally, the dose according to the patient's body weight is 0.001 mg / kg to 100 mg / kg, preferably 0.01 mg / kg to 30 mg / kg, and is preferably administered once to six times a day.

The pharmaceutical composition of the present invention may be used alone or in combination with methods for the prevention or treatment of lung cancer or using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers.

In addition, the present invention provides a health functional food for preventing or ameliorating lung cancer comprising sanggenol Q.

The health functional food for preventing or ameliorating lung cancer may be used as it is, or may be used together with other food or food ingredients, and suitably used according to a conventional method.

The term " functional food " as used herein means a food prepared and processed by using raw materials or ingredients having useful functions in the human body. The " function " Which may be useful for health use.

There is no particular limitation on the kind of the food. Examples of foods to which sanggenol Q can be added include various soups, beverages, tea, drinks, alcoholic beverages and vitamin complexes, and may include all health foods in a conventional sense. In addition to the above, sanggenol Q of the present invention can be used in various forms such as various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickening agents, Preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the health functional food of the present invention may contain flesh for the production of natural fruit juice, fruit juice drink and vegetable drink. These components may be used independently or in combination.

The pharmaceutical composition for prevention or treatment of lung cancer comprising sanggenol Q according to the present invention activates caspase 3 and caspase 9 in lung cancer cells, promotes poly (ADP ribose polymerase) cleavage, Bcl-2, XIAP and Bad, thereby having excellent lung cancer treatment and prevention effect.

1 is a graph showing lung cancer cell killing effect according to the concentration of sanggenol Q. Fig.
FIG. 2A shows the expression of apoptotic protein according to treatment with sanggenol Q, and FIG. 2B shows apoptosis following treatment with Sanguinol Q by TUNEL analysis.
FIG. 3 is a graph showing the expression of a gene related to a hedgehog signal according to the treatment of sanggenol Q. FIG.
FIG. 4 is a graph showing changes in cell and sub-G1 phase of apoptosis following treatment with sanggenol Q. FIG.

Hereinafter, the present invention will be described in detail by way of examples with reference to the drawings. However, the embodiments according to the present disclosure can be modified in various other forms, and the scope of the present specification is not construed as being limited to the embodiments described below. Embodiments of the present disclosure are provided to more fully describe the present disclosure to those of ordinary skill in the art.

<Extraction and Separation of Sangenol Q>

The dried powder of M. alba (mulberry) was extracted with 80% methanol at room temperature for 24 hours (68 L x 3). Filtered through filter paper, and concentrated via a vacuum rotary evaporator.

The remaining concentrate was suspended in 2 L of water and extracted successively with ethyl acetate (EtOAc, 2 L x 2) and normal-butanol (n-BuOH, 1.8 L x 3). The concentrate was partitioned into EtOAc fraction (MRE, 580 g), n-BuOH fraction (MRB, 114 g) and H ₂ O fraction (MRW, 1006 g) The EtOAc layer was separated into 41 layers, including Sanggenol Q, by silicon dioxide tube chromatography (MRE1 to MRE41).

&Lt; Example 1: Culture and cell viability of lung cancer cells >

Culture of lung cancer cells

Fetal serum (FBS), 2 mM glutamine, 100 U / mL penicillin and 100 μg / mL streptomycin were purchased from ATCC (American Type Culture Collection: Rockville, MD) the addition of RPMI 1640 medium and incubated in 5% C0 _2.

Cell viability analysis

For cell viability analysis, H460, A549 and H1299 cells were cultured in 70%, treated with different concentrations of sanggenol Q for 24 hours, and cell viability was measured using MTT assay Respectively.

FIG. 1 shows the killing effect of lung cancer cell lines H460, A549 and H1299 according to the concentration of sanggenol Q, and FIG. 2B shows the killing of H460, a lung cancer cell line, by confocal microscopy through TUNEL analysis. As shown in Fig.

As can be seen from Figs. 1 and 2, viability of the lung cancer cell line was decreased according to the concentration of sanggenol Q and the treatment time. The pharmaceutical composition for prevention or treatment of lung cancer including sanggenol Q has lowered the survival ability of lung cancer cells and has a therapeutic and preventive effect on lung cancer.

Example 2. Western blot analysis for protein determination < RTI ID = 0.0 >

After incubation, sanggenol Q was added and further incubated for 24 hours. Cells were washed with PBS and lysed (20 mM sucrose, 1 mM EDTA, 20 μM Tris-Cl, pH 7.2, 1 mM DTT, 10 mM KCl, 1.5 mM MgCl ₂ , 5 μg / mL pepstatin A, 10 μg / ml leupeptin and 2 μg / mL aprotinin). After centrifugation, the supernatant was collected and the protein concentration was measured. The aliquot containing 30 μg was separated on SDS-polyacrylamide gel using a bio-rad protein assay kit, and the indicated primary antibody was conjugated to the peroxidase conjugate 2 The ECL detection system was used to analyze PARP, caspase 3, 8, 9, bc1-2, XIAP, Bad, Shh, PTCH-1, PTCH-2, SUFU, GLI and actin using secondary antibodies.

2A shows the expression (2A) of the apoptosis protein according to the treatment with sanggenol Q by the Western blot method.

2 shows that cleaved caspase 3 and PARP associated with apoptosis are increased while that of procaspase 9, XLAP and Bad is decreased.

3 is a graph showing a change in the expression of a gene related to a hedgehog signal according to treatment with sanggenol Q. Fig. 3 shows that administration of sanggenol Q suppresses Shh and SUFU, which are genes involved in hedgehog signaling involved in cancer metastasis, while PTCH-1, PTCH-2, GLI could be confirmed to increase.

&Lt; Example 3: Cell cycle analysis >

After cell culture, trypsin-EDTA was removed and fixed with 75% ethanol. Ethanol was removed and RNase was treated for 1 hour. After the reaction, RNase was removed and Propidium Iodide (PI) was added to analyze the cell cycle using a flow cytometer.

4A is a graph showing cell changes of apoptosis upon treatment with sanggenol Q. Fig. As shown in FIG. 4A, when treated with sanggenol Q, it was confirmed that cells causing apoptosis were increased.

&Lt; Example 4: Apoptosis analysis >

After cell culture, sanggenol Q was treated with Annexin V compound for 24 hours, treated with FITC, and DAPI solution was added at 1 mg / ml and analyzed by flow cytometry.

FIG. 4B is a graph showing a change in the Sub-G1 phase when the cell cycle is analyzed according to the treatment with sanggenol Q. FIG. As a result of FIG. 4B, it was confirmed that the treatment with sanggenol Q significantly increased the sub-G1 population when treated with sanggenol Q (left side of FIG. 4B) .

The above results indicate that treatment with sanggenol Q significantly activates apoptosis of lung cancer cell lines, decreases the expression of Survival protein, significantly increases the expression of cell death proteins . Therefore, it can be confirmed that a pharmaceutical composition containing sanggenol Q has an advantageous effect on prevention and treatment of lung cancer.

Claims (5)

A pharmaceutical composition for preventing or treating lung cancer comprising sanggenol Q. The method according to claim 1,
Wherein the sanggenol Q is extracted and isolated from morus alba. The method according to claim 1,
Wherein the prevention or treatment of lung cancer is by promoting the death of lung cancer cells. The method according to claim 1,
Wherein the lung cancer is small cell lung cancer (SCLC) or non-small cell lung cancer (NSCLC). A health functional food for prevention or improvement of lung cancer including sanggenol Q.

Images