KR101978673B1 - Composition for prevention, improvement or treatment of liver cancer or stomach cancer comprising sinensetin as active ingredient - Google Patents
Composition for prevention, improvement or treatment of liver cancer or stomach cancer comprising sinensetin as active ingredient Download PDFInfo
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- KR101978673B1 KR101978673B1 KR1020170146044A KR20170146044A KR101978673B1 KR 101978673 B1 KR101978673 B1 KR 101978673B1 KR 1020170146044 A KR1020170146044 A KR 1020170146044A KR 20170146044 A KR20170146044 A KR 20170146044A KR 101978673 B1 KR101978673 B1 KR 101978673B1
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- cancer
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- cisplatin
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- 208000005718 Stomach Neoplasms Diseases 0.000 title abstract description 38
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/28—Compounds containing heavy metals
- A61K31/282—Platinum compounds
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- A—HUMAN NECESSITIES
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- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/308—Foods, ingredients or supplements having a functional effect on health having an effect on cancer prevention
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Abstract
The present invention relates to a composition for preventing, ameliorating or treating liver cancer or stomach cancer containing cinnensetin as an active ingredient, and more particularly to a composition for preventing, ameliorating or treating liver cancer or stomach cancer containing cinnensetine or cinnenecetin and cisplatin as an active ingredient Therapeutic and transfection-inhibiting pharmaceutical compositions, functional food compositions and functional feed compositions.
The composition of the present invention can effectively prevent, ameliorate, or treat liver cancer or stomach cancer by inducing the death of liver cancer cells or stomach cancer cells without significantly affecting normal cells by containing cinnenecetin as an active ingredient, It is possible to effectively inhibit metastasis of liver cancer or stomach cancer by inhibiting mobility. In particular, a composition containing cinnensetin as an active ingredient together with cisplatin is more effective in preventing gastric cancer, preventing or ameliorating gastric cancer, and inhibiting metastasis, because of the synergistic effect of the two components, have.
Description
The present invention relates to a composition for preventing, ameliorating or treating liver cancer or stomach cancer containing cinnensetin as an active ingredient, and more particularly to a composition for preventing, ameliorating or treating liver cancer or stomach cancer containing cinnensetine or cinnenecetin and cisplatin as an active ingredient Therapeutic and transfection-inhibiting pharmaceutical compositions, functional food compositions and functional feed compositions.
Cancer refers to a group of cells that are caused by continuous division and proliferation of abnormal cells by destroying the balance of cell division and death by various causes, and are sometimes referred to as tumors. It usually occurs in more than 100 different parts of the body, including lungs, lymph nodes, bones, and white blood cells, and has a high mortality rate due to metastasis to the surrounding tissues. The highest cancer death rate in Korea in 2015 (deaths per 100,000 population) was gastric cancer in the 30s and liver cancer in the 40s and 50s. Several strategies for treating this are being studied.
There are radiation, chemotherapy, and surgical treatments for cancer treatment, but many side effects have been reported. Cisplatin is an anticancer drug used for the treatment of various types of solid cancer such as stomach cancer, colon cancer, liver cancer and lung cancer. It is the most common substance used alone or in combination with other anticancer drugs for cancer treatment. However, side effects such as vomiting due to cisplatin, diarrhea, cystitis, and renal dysfunction have been reported. Therefore, a new approach is needed for cancer treatment and adjuvant treatment. In this regard, research has been conducted to develop a therapeutic agent having a low side effect and excellent anticancer effect by using natural substances relatively low in toxicity compared to the synthesized substance, and to develop a substance capable of reducing the adverse effects of existing anticancer drugs and increasing their effects.
Flavonoids in natural products have two phenyl rings and a heterocyclic ring and have 15 carbon skeletons. Extracts and single components of flavonoids have been reported to have a wide range of pharmacological and biological activities (antiallergic, antiinflammatory, antioxidant, antimicrobial, anti-cancer, etc.). One of the single components of flavonoids, methylated flavone, methylenated flavone, is contained in a large amount in orange oil, and anti-inflammatory and antiviral effects have been reported.
The present inventors have studied a method for overcoming the problems of the conventional anticancer drugs by using cinnensetine as one of such flavonoid components. As a result, the present inventors have found that the above- Or stomach cancer cells can be effectively killed and the mobility of these cells can be effectively inhibited. In particular, when used in combination with the existing anticancer drug cisplatin, synergistic effects were shown in the killing effect and the mobility inhibition effect of gastric cancer cells, and it was confirmed that the use of cisplatin could be reduced to reduce the side effects and increase the anti- Thereby completing the invention.
Accordingly, it is a main object of the present invention to provide a composition having few side effects using a natural substance and excellent in prevention, improvement or therapeutic effect of cancer.
It is another object of the present invention to provide a composition having a superior effect of inhibiting cancer metastasis using a natural substance substance.
According to one aspect of the present invention, there is provided a pharmaceutical composition for the prophylaxis or treatment of liver cancer or stomach cancer containing sinensetin or a salt thereof as an active ingredient.
In the pharmaceutical composition for preventing or treating liver cancer or stomach cancer of the present invention, it is preferable to further contain cisplatin as an active ingredient in addition to the above-mentioned sinensetin or a salt thereof.
According to another aspect of the present invention, there is provided a functional food composition for preventing, ameliorating or preventing the transplantation of liver cancer or stomach cancer containing sinensetin or a salt thereof as an active ingredient.
In the functional food composition for preventing, ameliorating or inhibiting metastasis of liver cancer or stomach cancer of the present invention, it is preferable to further contain cisplatin as an active ingredient in addition to the above-mentioned sinensetin or a salt thereof.
According to another aspect of the present invention, there is provided a functional feed composition for preventing, ameliorating or preventing the transplantation of liver cancer or stomach cancer containing sinensetin or a salt thereof as an active ingredient.
In the functional feed composition for preventing, ameliorating or inhibiting metastasis of liver cancer or gastric cancer according to the present invention, it is preferable to further contain cisplatin as an active ingredient in addition to the above-mentioned sinensetin or a salt thereof.
According to still another aspect of the present invention, there is provided a pharmaceutical composition for inhibiting the metastasis of liver cancer or stomach cancer containing sinensetin or a salt thereof as an active ingredient.
In the pharmaceutical composition for inhibiting metastasis of liver cancer or stomach cancer according to the present invention, it is preferable to further contain cisplatin as an active ingredient in addition to the above-mentioned sinensetin or a salt thereof.
The composition of the present invention can effectively prevent, ameliorate, or treat liver cancer or stomach cancer by inducing the death of liver cancer cells or stomach cancer cells without significantly affecting normal cells by containing cinnenecetin as an active ingredient, It is possible to effectively inhibit metastasis of liver cancer or stomach cancer by inhibiting mobility. In particular, a composition containing cinnensetin as an active ingredient together with cisplatin is more effective in preventing gastric cancer, preventing or ameliorating gastric cancer, and inhibiting metastasis, because of the synergistic effect of the two components, have.
FIG. 1 shows the results of experiments on the killing effect of cinnenecetin (Si) on HepG2 cells. Cell viability was measured by MTT assay, and HepG2 cells were treated with 25 ~ 100 μM of cinnenecetin for 24 h and 48 h; #, p < Thle2 cell control; ## p <0.01 vs. Thle2 cell Control; *, p < HepG2 Cell control; ** p < HepG2 cell Control; *** p < HepG2 cell Control.
FIG. 2 shows the results of experiments on the killing effect of AGS cells of cinnenecetin (Si). Treatment of cinnenecetin with AGS cells at a concentration of 20-80 μM for 24 h; * p <Control; ** p <Control; *** p < Control.
Fig. 3 shows the results of experiments on the killing effect of AGS cells of cisplatin (Cis). Treatment of AGS cells with cisplatin at a concentration of 0.1-0.3 μM for 24 h; ** p < Control.
FIG. 4 shows the results of an experiment on the killing effect of AGS cells when cisplatin (Cis) and synenecetin (Si) were treated at the same time. AGS cells were treated with cisplatin at a concentration of 0.1, 0.2 μM, cinnexetin at 40, 50 μM for 24 h; ** p <Control; *** p <Control;### p <0.001 vs. Cis 0.1 [mu] M or Cis 0.2 [mu] M; $$ p <
FIG. 5 shows the results of an experiment for inhibiting the mobility of HepG2 cells of cinnenecetin (Si). Wound healing assay, analysis of cell area after treatment with cynesetin at 50, 100 μM for 48 h; *** p < Control.
Fig. 6 shows the results of experiments on the effect of inhibiting the migration of AGS cells of cisplatin (Cis) and cinnenecetin (Si). Cisplatin and cinnenecetin were measured at 24 h after treatment at each concentration; * p <Control; *** p <Control;# p < Cis 0.1 [mu] M or Cis 0.2 [mu] M; $$ p <
The composition of the present invention contains sinensetin or a salt thereof as an active ingredient or a composition containing sinensetin or a salt thereof; And cisplatin or a salt thereof as an active ingredient.
The chemical structure of cinnenecetin is shown in the following chemical formula 1. Cynenecetine is a methylated flavone, which is found in a lot of orange oil, and has been reported to have anti-inflammatory and antiviral effects. In the present invention, by disclosing a new physiological activity, the present invention provides the use of prevention, improvement or treatment of liver cancer or stomach cancer and the use of inhibiting the metastasis of liver cancer or stomach cancer.
[Chemical Formula 1]
The chemical structure of cisplatin is represented by the following chemical formula 2. It is used for the treatment of various types of cancer such as stomach cancer, colon cancer, liver cancer, and lung cancer. However, there are side effects such as vomiting, diarrhea, cystitis, and kidney dysfunction.
(2)
According to the present invention, cinnenecetin can kill hepatocellular carcinoma cells or stomach cancer cells without significantly affecting normal cells, and can inhibit the mobility of liver cancer cells or stomach cancer cells.
In addition, when cinnenecetin is used together with cisplatin, the synergistic effect can be exhibited in the killing effect and the mobility inhibition effect of gastric cancer cells.
Based on these effects, cinnenecetin can be used as an anticancer adjuvant to increase the anticancer effect of cisplatin or as an antitumor agent to increase the suppression effect of metastasis.
It is believed that the salts of cinnenecetin in the present invention may be used in the form of salts, such as acid, pharmacologically or pharmacologically acceptable acid addition salts or metal complexes, such as zinc and iron.
The pharmaceutical composition of the present invention can be administered orally or parenterally at the time of clinical administration, and can be administered orally or parenterally at the time of parenteral administration, and can be administered intraperitoneally, rectally, subcutaneously, intravenously, intramuscularly, Or intramuscular injection, and may be used in the form of a general pharmaceutical preparation.
The pharmaceutical compositions of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers.
The daily dosage of the pharmaceutical composition of the present invention may be about 1 to 100 mg / kg on an adult basis per day based on the cinnenecetin or a salt thereof of the present invention contained in the composition, and may be administered once to several times a day The range may vary depending on the patient's weight, age, sex, health condition, diet, time of administration, administration method, excretion rate, and severity of the disease. The cisplatin may be administered at a level that is used for the treatment of existing gastric cancer, and according to the present invention, the combination with cinnenecetine may be administered at a lower level because the anticancer effect is enhanced. For example, the adult standard may be about 1 to 50 mg / kg per day.
In practical administration, it may be administered in various formulations of parenteral administration. In the case of formulation, it may be prepared by using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants and the like. Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, etc. may be used as the non-aqueous solvent and suspension agent. The base of suppositories may be witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.
The pharmaceutical composition of the present invention may contain one or more active ingredients showing the same or similar functions in addition to the cinnenecetin or a salt thereof of the present invention or cisplatin.
The food composition of the present invention may be a composition based on the effective amount of cinnenecetin or a salt thereof, or a mixture of cisplatin itself or a pharmaceutically acceptable carrier. The food composition of the present invention can be applied to food products such as meat products, fish products, tofu, mushrooms, porridge, noodles such as noodles or noodles, seasonings such as soy sauce, miso, It is considered that food types such as pickles such as pickles, pickles, fruits, vegetables, soymilk, and fermented beverages are possible. The pharmacologically acceptable carrier may also be the pharmaceutically acceptable carrier.
The feed composition of the present invention may be a composition based on the effective amount of cinnenecetin or a salt thereof, or a composition obtained by mixing cisplatin itself or a dietary acceptable carrier. The feed composition of the present invention is considered to be in the form of compound feed, pellet, and silage. The compound feed can be prepared by mixing various kinds of general feeds and the above-mentioned cinnenecetin or a salt thereof, or cisplatin. Feeds in the form of a pellet may be prepared by formulating the compound feed into a pelletizer, which may be prepared by fermenting a chrysanthemum and then adding the cinnenecetin or a salt thereof, or cisplatin.
Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.
[Example]
1. Materials and Methods
1-1. reagent
Sinensetin was purchased from MedChem Express (MCE, USA) and stocked with DMSO at 10 mM. Dulbecco's Modified Eagles medium (DMEM) used for cell culture was purchased from Hyclone (Logan, UT), and Bronchial Epithelial Cell Growth Medium (BEGM ™ ) Bulletkit ™ was purchased from Lonza (Switzerland). Penicillin / streptomycin and fetal bovine serum (FBS) added to the medium were purchased from Gibco (Grand Island, NY). MTT (3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide) used for the measurement of apoptosis was purchased from Sigma-Aldrich (St. Louis, Mo.).
1-2. Cell culture and sample treatment
HepG2 cells, which are human liver-derived hepatic cancer cells, were cultured in DMEM medium. AGS cells derived from human cells were cultured in RPMI medium supplemented with 10% FBS and 1% penicillin / streptomycin. And cultured in a CO 2 incubator (5% CO 2 , 95% air) at 37 ° C. When the cells were over 80% in the culture dish, the cells were washed with phosphate buffered saline (PBS, pH 7.4) and subcultured with 0.25% trypsin-2.65 mM EDTA. Cynecetin was added to 10 mM of DMSO and treated with 50 μM and 100 μM of HepG2 cells. AGS cells were treated with 40 μM and 50 μM of cinnenecetin with 0.1 μM and 0.2 μM of cisplatin, and the same amount of DMSO was added to the control.
1-3. Measurement of cytotoxicity (MTT assay)
HepG2 and AGS cells were plated in 48-well plates at 5 × 10 4 and incubated for 16 h at 37 ° C in a 5% CO 2 incubator. After the medium was removed, each of the following samples was treated according to the concentration, and the control group was treated with dimethyl sulfoxide (DMSO). Each sample was treated with 50 μl of MTT (0.05 g / 10 ml; 0.5%) solution (final 0.05% MTT) for 48 hours for HepG2 and for 24 hours for AGS. The cells were reacted for 3 hours in a 5% CO 2 incubator at 37 ° C to reduce MTT. All wells were removed and 500 ul of DMSO was added to each well. The absorbance was measured at 560 nm using an ELISA reader.
1-4. Migration assay
HepG2 and AGS cells were plated at 2 × 10 5 in a 12-well plate for cell culture and cultured for 16 hours. The cells were scrunched using a plastic 200ul tip and washed with PBS. After that, the drug was treated at each concentration, and the cells were reacted in CO 2 incubator for each hour. The cell mobility was photographed and the area was measured using the Image J program.
1-5. Cell death (Annexin V measurement)
HepG2 and AGS cells were plated at 5 × 10 5 in a 6-well plate and treated with each concentration 16 hours later. After the reaction time of the drug has elapsed, cells were washed once with PBS and cells were collected using typsin-EDTA. Cells were stained with FITC Annexin V Apoptosis Detection Kit I (BD Biosciences, San Jose, Calif.) By flow cytometry using FACS Calibur (BD Biosciences).
1-6. Cell cycle analysis (PI staining)
HepG2 and AGS cells were plated in 6 well plates and flow cytometry was used. HepG2 and AGS cells were collected after each reaction time, and cells were fixed at -20 ℃ for 1 hour with 70% ethanol. The cells were washed with PBS and reacted with 1 U / ml of RNase A (DNase free) and PBS at 37 ° C for 1 hour, then 5 ug / ml of propidium iodide (PI) And then reacted. 10,000 cells per sample were measured using a FACS Calibur (BD Biosciences, San Jose, Calif.).
1-7. Protein expression experiments (Western blot analysis)
HepG2 cells treated with cinnenecetin were washed with PBS and incubated with RIPA lysis buffer (50 mM Tris-HCl pH 7.4, 1% NP-40, 0.25% Na-deoxycholate , 150 mM NaCl, 1 mM EDTA) was added to extract whole proteins of the cells. Bicinchoninate (BCA) assay was used to quantitate the protein concentration, electrophoresed on SDS-polyacrylamide gel, and then transferred to the PVDF membrane. To prevent the binding of nonspecific antibodies to membranes, the cells were incubated in 5% skim milk for 1 hour. After incubation at 4 ° C with primary antibody, horse radish peroxidase (HRP) conjugated secondary antibody was reacted. The antibodies were reacted with ECL kit (Promega, USA) and then measured using Chemidoc instrument (Bio Rad, California, USA).
1-8. Statistical processing
All experimental results were repeated at least 3 times and the results were expressed as mean ± standard deviation. The statistical significance of each test result was determined by Student's t-test as compared to the control group without sample and statistically significant when the P value was less than 0.05.
2. Results
2-1. Induction of HepG2 cell death of cinnenecetin
One of the flavonoids, cinnexetin, was treated with time and concentration to determine the death of HepG2, a human liver cancer cell, and MTT assay was performed. HepG2, a human liver cancer cell, showed apoptosis in a time - and concentration - dependent manner in groups treated with 24 hours and 48 hours of cinnenecetine. When compared with the control group, the cancer cell killing activity was confirmed to be about 50% at 50 μM and 65% at 100 μM in the group treated with cinnensetin for 48 hours. In addition, in order to confirm whether it affects extrahepatic extra-cellular cells, the human normal hepatocyte, Thle2 cells, was also treated with cinnenecetin. As a result, it was confirmed that cinnenecetin had a greater effect on liver cancer cells than hepatic normal cells in the group treated for 24 hours and 48 hours (see FIG. 1).
2-2. Effect of Cynenecetin on Cell Death in Gastric Cancer Cells
To determine whether AGS cells die in human gastric cancer cells, various concentrations of cinnenecetin were treated for 24 hours and then assayed by MTT assay. Cynenecetin showed AGS cell killing activity in a concentration dependent manner. When compared to the control group treated with DMSO, the cytotoxic effect was about 25% at 60 μM of cynesecetin and 35% at 80 μM (see FIG. 2). In addition, the cytotoxic effect of cisplatin, which is known as an anticancer drug, was examined in AGS cells, and showed a killing effect of about 50% at 0.3 μM of cisplatin (see FIG. 3).
2-3. Increased cell killing effect by combination of cinnenecetin and cisplatin
AGS cells were treated with 40 μM and 50 μM of cinnenecetine and treated with 0.1 μM and 0.2 μM each cisplatin. When treated with 0.1 μM of cisplatin alone, cell viability was reduced to about 30% in the group treated with 50 μM of cinnenecetin, although the cell viability was about 90% (see FIG. 4). In addition, about 75% cell viability was observed in the group treated with 0.2 μM of cisplatin alone, but about 50% cell viability was shown when treated with 40 μM of cinnenecetin (see FIG. 4). Thus, when cisplatin and cinnenecetin are treated together, cisplatin exhibits a higher apoptotic action in AGS cells, which are human gastric cancer cells.
2-4. Identification of HepG2 and AGS cell migration effect of cinnenecetin
Cell migration assay (wound healing assay) was performed to investigate the effect of cinnenecetin on HepG2 cell mobility. When HepG2 was treated with 50 μM and 100 μM of Sinensetin for 48 h, the cell migration area was 50% at 50 μM and 30% at 100 μM, compared with the control. As a result, it was confirmed that the mobility of the cells was inhibited by cinnenecetin in a concentration-dependent manner of HepG2 cells (see FIG. 5). In the case of AGS cells, cinnenecetin and cisplatin inhibited cell mobility compared to the control, respectively, but the combination of cinnenecetin and cisplatin showed greater mobility inhibition (see FIG. 6).
Claims (8)
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| KR20230022792A (en) | 2021-08-09 | 2023-02-16 | 주식회사 네오나 | Composition comprising a MSH2 inhibitor for preventing or treating cancer |
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