CN109640978B - Pharmaceutical composition containing apigenin, curcumin and honokiol as effective components for preventing or treating lung cancer - Google Patents
Pharmaceutical composition containing apigenin, curcumin and honokiol as effective components for preventing or treating lung cancer Download PDFInfo
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Abstract
The invention relates to a pharmaceutical composition for preventing or treating lung cancer, which takes any one of apigenin and curcumin and honokiol as active ingredients. The composition according to the present invention can induce apoptosis of cancer cells in a mutually ascending manner when treated in combination or mixed, as compared with the case where apigenin, curcumin or honokiol is treated alone. The pharmaceutical composition for preventing or treating lung cancer, which contains any one of apigenin and curcumin and a honokiol mixture as active ingredients according to the present invention, has an improved anticancer effect, can induce apoptosis of cancer cells, and exhibits a very excellent cancer treatment enhancing effect. Therefore, the composition according to the present invention can be effectively applied to the prevention, amelioration or treatment of lung cancer.
Description
Technical Field
The invention relates to lung cancer treatment effect of any one of apigenin or curcumin and a honokiol mixture.
Background
Cancer (cancer) refers to an abnormal cell population generated by a balance between cell division and apoptosis due to various causes, by continuous division and proliferation, and is also called malignant tumor. In general, cancer is developed in a plurality of sites of more than 100 kinds of bodies including organs, leukocytes, bones, lymph nodes, and the like, and it develops into serious symptoms by a phenomenon of infiltrating peripheral tissues and metastasis to other organs, and finally, it causes death. Among them, lung cancer, which is a malignant tumor originated from the lung, is roughly classified into small cell lung cancer (small cell lung cancer) and non-small cell lung cancer (non-small cell lung cancer) according to its tissue morphology. The small cell lung cancer is classified into a part of lung cancer according to the location of the diseased tissue, but is clearly distinguished from other lung cancer in terms of clinical history, treatment method, prognosis, and the like, and thus is classified as described above. Non-small cell lung cancer is classified into adenocarcinoma, squamous cell carcinoma, and large cell carcinoma according to the tissue type.
Specifically, small cell lung cancer is basically large with a pale-white mass that proliferates along the bronchial wall. Small cell lung cancer has been developed, for the most part, at the time of diagnosis to such an extent that it is difficult to remove by surgery, is highly malignant, and thus rapidly grows, and is easily transferred to the whole body through lymphatic vessels or blood circulation. However, significant therapeutic effects can be obtained by chemotherapy or radiotherapy. The main organs of the small cell lung cancer metastasis are reported to be brain, liver, bone, lung, adrenal gland, kidney and the like, and mainly develop in the trachea (bronchus or bronchiole) first.
In contrast, as described above, non-small cell lung cancers are classified into adenocarcinoma (adenocarinoma), squamous cell carcinoma (squamous cell carcinoma), and large cell carcinoma (large-cell carcinoma). First, adenocarcinoma occurs mainly in the peripheral lung region, and often also in women or non-smokers, and is small in size but has metastasized. Recently, the frequency of the onset of the disease has been increasing. Secondly, squamous cell carcinoma is found mainly in the central part of the lung and in most cases grows into the lumen of the bronchi, resulting in the appearance of symptoms of obstruction of the bronchi. The onset of disease in men is common and closely related to smoking. Finally, large cell carcinoma occurs mainly near the lung surface (lung periphery), approximately half in the large bronchi, accounting for about 4-10% of all lung cancers, and the cell size is generally large. Some of them have a tendency to proliferate rapidly and metastasize, and large cell carcinoma has a poor prognosis compared to other non-small cell lung cancers.
If the lung cancer is diagnosed at an early stage, it can be recovered by medication, radiation therapy, or the like, but if the disease condition is deteriorated to a certain extent, the disease can be removed by surgery, and then the lung cancer can be treated by medication, radiation therapy, or the like. To date, the number of accepted drugs available for the treatment of lung cancer is limited to a few, so that it is difficult to perform a treatment suitable for a patient, and side effects thereof are also a big problem. Metastasis of cancer has been found mostly at the 1st observation of tumors, with low success rates of treatment after surgery.
Therefore, as a new approach to cancer treatment, efforts have been made to develop carcinogenic inhibitors and cancer therapeutic agents that have a low toxicity and a high effect of inhibiting cancer cell metastasis or reducing cancer cells, using natural substances having low toxicity. However, the fundamental disadvantage of poor therapeutic effects of these natural-derived therapeutic agents has not been solved (Korean patent laid-open No. 10-2012-0109139).
Under such a background, the present inventors have finally confirmed apigenin or curcumin through hard studies in order to develop a preparation capable of treating lung cancer more safely and effectively; and a honokiol mixture which can cause apoptosis of lung cancer cell lines and can inhibit proliferation of cancer cells, and which is found to be effective as a composition for treating lung cancer, thereby completing the present invention.
Disclosure of Invention
Technical problem to be solved
The present invention has been developed to solve the above problems. The inventors confirmed that the glycolysis rate of lung cancer cell lines is decreased, ANT2 expression is inhibited, HK2 expression is inhibited, PD-L1 expression is inhibited, and ATP production is reduced by administering apigenin, curcumin, and honokiol singly or in combination. Based on this, the present invention has been completed.
Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating lung cancer, which comprises one of apigenin and curcumin, and honokiol as active ingredients.
Another object of the present invention is to provide a health functional food composition for preventing or improving lung cancer, which comprises one of apigenin and curcumin, and honokiol as active ingredients.
However, the technical problems to be solved by the present invention are not limited to the above-mentioned problems, and other problems not yet mentioned will be apparent to those skilled in the art from the following description.
Means for solving the problems
In order to achieve the above objects of the present invention, the present invention provides a pharmaceutical composition for preventing or treating lung cancer, which comprises one of apigenin and curcumin, and honokiol as active ingredients.
Another object of the present invention is to provide a health functional food composition for preventing or improving lung cancer, which contains one of apigenin and curcumin and honokiol as active ingredients.
As an embodiment of the present invention, the composition comprises apigenin and honokiol as effective ingredients.
As another embodiment of the present invention, the composition comprises curcumin and honokiol as effective ingredients.
As still another embodiment of the present invention, the composition may reduce the expression of ANT2(Adenine nucleotide transporter 2; Adenine nucleotide transporter 2) gene.
As yet another example of the present invention, the composition may decrease the expression of HK2(hexokinase 2; hexokinase 2) or PD-L1(Programmed death ligand 1; Programmed death-ligand 1) protein.
In addition, the present invention provides a method for preventing or treating lung cancer comprising the step of administering the pharmaceutical composition to a subject.
In addition, the invention provides a use of the pharmaceutical composition for preventing or treating lung cancer.
ADVANTAGEOUS EFFECTS OF INVENTION
The invention relates to a compound of apigenin or curcumin; and honokiol as effective component for preventing or treating lung cancer. According to the composition of the present invention, cancer cell apoptosis can be induced mutually ascendingly if it is mixed or combined with treatment, compared with the case where apigenin, curcumin or honokiol is treated alone. The pharmaceutical composition for preventing or treating lung cancer, which comprises any one of apigenin and curcumin and a honokiol mixture as active ingredients, can improve the anti-cancer effect and induce cancer cell apoptosis, thereby showing a very good cancer treatment augmentation effect. Therefore, the composition according to the present invention can be effectively used for the prevention, amelioration or treatment of lung cancer.
Drawings
FIG. 1 shows the results of confirmation of apoptosis effect of lung cancer cell lines when honokiol, apigenin and curcumin are treated alone or in combination;
FIG. 2 shows the results of examining the effect of inhibiting the expression of ANT2 in lung cancer cell lines by treating honokiol, apigenin, and curcumin alone or in combination;
FIG. 3 shows the results of confirmation of the effect of reducing ATP production in lung cancer cell lines when honokiol, apigenin, and curcumin are treated alone or in combination;
FIG. 4 shows the results of confirming the inhibitory effect of HK2 expression in lung cancer cell lines when honokiol, apigenin, and curcumin were treated alone or in combination;
FIG. 5 shows the results of confirming the effect of reducing the glycolysis rate (glycolysis rate) of lung cancer cell lines when honokiol, apigenin, and curcumin are treated alone or in combination;
FIG. 6a shows the results of the inhibition of PD-L1 expression in lung cancer cell line A549 when honokiol, apigenin and curcumin were treated alone or in combination;
FIG. 6b shows the results of the inhibition of PD-L1 expression in lung cancer cell line H2228 when honokiol, apigenin, and curcumin were treated alone or in combination.
Detailed Description
The composition of the present invention contains apigenin or a mixture of curcumin and honokiol as active ingredients, and has been confirmed to have lung cancer prevention, improvement or treatment effects by reducing glycolysis rate of lung cancer cell lines, inhibiting ANT2 expression, inhibiting HK2 expression, inhibiting PD-L1 expression and reducing ATP production, and thus the present invention has been completed based on this.
Hereinafter, the present invention will be described in detail.
The invention provides a compound of apigenin or curcumin; and honokiol as an active ingredient, and a composition for preventing or treating lung cancer containing the same. The composition comprises a pharmaceutical and health functional food composition.
The term "prevention" as used in the present invention means that all behaviors of lung cancer-related diseases are inhibited or onset is delayed by administering the pharmaceutical composition according to the present invention.
The term "treatment" as used in the present invention refers to all actions that improve the symptoms of lung cancer or become beneficial by administering the pharmaceutical composition according to the present invention.
The disease to be prevented or treated by the composition of the present invention, namely "cancer (cancer)" is a disease that causes malignant tumor due to unlimited proliferation of cells in a biological tissue, and more specifically, may be lung cancer, but is not limited thereto.
The "honokiol" used in the present invention is a polyphenol compound which can be isolated, purified or extracted from Magnoliaceae species (Magnolia species). More specifically, it can be isolated, purified or extracted from magnolia officinalis, but is not limited thereto, and a chemically synthesized compound or a commercially available compound can be used. In addition, the derivative or the analogue of honokiol can be used, but the invention is not limited to the derivative or the analogue.
The "apigenin (5, 7,4 '-trihydroxyflavone; 5,7,4' -trihydroxyflavone)" used in the present invention is a flavonoid-series compound, which can be found in not only the extract of robinia pseudoacacia but also a large amount of a compound that is present in fruits such as orange, apple, cherry, and grape, vegetables such as onion, caraway, celery, barley, and tomato, or beverages such as tea water and wine, and can be separated, purified, or extracted therefrom, but is not limited thereto, and a compound synthesized by a chemical method or a commercially available compound can be used. Further, a derivative or an analog of apigenin may be used, but the present invention is not limited thereto.
"curcumin (curcumin)" used in the present invention is an alkaloid compound, and can be isolated, purified or extracted from turmeric or curcuma aromatica, but is not limited thereto, and a compound synthesized by a chemical method or commercially available can be used. In addition, derivatives or analogs of curcumin may be used, but not limited thereto.
The composition of the present invention may be a composition for preventing, improving or treating lung cancer, which contains apigenin and honokiol as active ingredients, but is not limited thereto.
The composition of the present invention may be a composition for preventing, improving or treating lung cancer, which contains curcumin and honokiol as active ingredients, but is not limited thereto.
In addition, the composition according to the present invention is characterized in that the expression of ANT2 (adenosine nucleotide translocator 2) gene is reduced.
The "ANT 2 (adenosine nucleotide translocator 2)" of the present invention is a gene having an important function in the survival, proliferation and metastasis processes of cancer cells. The adenine nucleotide translocator/transporter (ANT)2 protein is one of the most abundant proteins in the inner mitochondrial membrane, formed as six transmembrane helices. ANT promotes the conversion of mitochondrial ATP to cytoplasmic ADP, playing an important role in the process of cellular energy metabolism. This has an effect on mitochondrial oxidative phosphorylation. All mammals carry isoforms (isoforms) of ANT 1-ANT 3, and the conversion of ANT2, which is a family of isoforms, to intramitochondrial ADP increases cytoplasmic ATP uptake by glycolysis process, thereby enabling maintenance of mitochondrial membrane potential, and the deletion (knockdown) of ANT2 reduces membrane potential difference in cancer cells.
Furthermore, the composition according to the present invention is characterized in that the expression of HK2(hexokinase 2) or PD-L1(Programmed death-ligand 1) protein is reduced.
The "HK 2(hexokinase 2)" of the present invention is an enzyme that converts hexose having 6 carbons into hexose phosphate by phosphorylation treatment, and although HK2 is expressed in a small amount in normal cells, it is expressed in a very high amount in most cancer cells, and has a function of phosphorylating glucose, and at the same time, binds to mitochondrial VDAC protein and inhibits Apoptosis mechanism (Apoptosis), and has a function of enhancing the viability of cancer cells. Therefore, it is useful for cancer treatment targeting HK 2.
In addition, the "PD-L1 (Programmed death-ligand 1)" protein of the present invention is a protein present on the surface of cancer cells, and when T cells having a cancer cell-killing effect bind to PD-L1, which is a protein present on the surface of cancer cells, the T cells cannot attack the cancer cells and the cancer cell-killing effect is lost. The immunoanti-cancer agent prevents the antibody from binding to the PD-L1 protein, thereby allowing the T cell to actively attack the cancer cell. The anticancer agent can enhance human immune system, and has no side effect or tolerance caused by existing anticancer agent. Therefore, as one of the immune avoidance strategies of cancer cells, the function of tumor-specific T lymphocytes can be inhibited by modulating the expression of PD-L1 and utilizing the change in the function of immune checkpoint (immune checkpoint).
In one embodiment of the present invention, the apoptosis effect of cancer cells was confirmed by treating lung cancer cell lines with honokiol, apigenin, and curcumin alone or in combination (see example 1). Meanwhile, the anti 2 expression inhibitory effect (see example 2), ATP production reducing effect (see example 3), HK2 expression inhibitory effect (see example 4), glycolysis rate reducing effect (see example 5), and PD-L1 expression inhibitory effect (see example 6) of lung cancer cell lines were confirmed when honokiol, apigenin, and curcumin were treated alone or in combination.
Therefore, the pharmaceutical composition containing either apigenin or curcumin and honokiol as active ingredients according to the present invention can reduce glycolysis rate of lung cancer cell lines, inhibit ANT2 expression, inhibit HK2 expression, inhibit PD-L1 expression and reduce ATP production, and can also be used as an active ingredient of a composition for preventing, improving or treating lung cancer.
The pharmaceutical composition according to the present invention may further include a pharmaceutically acceptable carrier in addition to the active ingredient. In this case, the pharmaceutically acceptable carrier is a substance commonly used in formulation of a preparation, and includes, but is not limited to, lactose, glucose, sucrose, sorbitol, mannitol, starch, locust bean gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methyl paraben, propyl paraben, talc, magnesium stearate, mineral oil, and the like. In addition, in addition to the above components, a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like may be further included.
The pharmaceutical composition of the present invention may be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically) depending on the intended method, and the dose varies depending on the condition and body weight of the patient, the degree of disease, the form of the drug, the route of administration and the time, but can be appropriately selected by those skilled in the art.
The pharmaceutical compositions of the present invention are administered in a pharmaceutically effective amount. In the present invention, the "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the standard of effective amount may be determined according to factors including the kind of disease, the severity, the activity of a drug, the sensitivity to a drug, the administration time, the administration route and the excretion ratio of a patient, the treatment period, the concurrent use of a drug, and other factors well known in the medical field. The pharmaceutical compositions according to the invention may be administered either as a single therapeutic agent or in combination with other therapeutic agents, either sequentially or simultaneously with existing therapeutic agents, either singly or multiply. It is important that the administration be carried out on a dosage level that achieves the maximum effect with the minimum amount, taking all of the above factors into consideration and without producing side effects, as can be readily determined by one skilled in the art.
Specifically, the effective amount of the pharmaceutical composition of the present invention varies depending on the age, sex, condition, body weight, absorption rate, inert rate and excretion rate of the active ingredient in vivo of the patient, the kind of the disease, and the drugs used in combination. Generally, the administration is carried out in a standard dose of 0.001 to 150mg, preferably 0.01 to 100mg, per 1kg body weight, daily or every other day, or 1 to 3 divided doses per 1 day. However, it may be increased or decreased depending on the administration route, severity of obesity, sex, body weight, age, etc. Therefore, the method of administration of the dose is not intended to limit the scope of the present invention.
In addition, the composition of the present invention can be added to a health functional food for the purpose of preventing or improving lung cancer-related diseases. When the compound having a lung cancer preventive or ameliorating effect of the present invention is used as a dietary supplement, the compound may be added directly or used together with other foods or food ingredients, and may be used as appropriate according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use (prophylactic, health or therapeutic treatment). In general, the compound of the present invention is added in an amount of 15% by weight or less, preferably 10% by weight or less, relative to the raw material at the time of producing a food or drink. However, when the intake is performed for a long period of time for the purpose of health and hygiene or for the purpose of health regulation, the amount may be in the range or less. From the viewpoint of safety, there is no problem, and therefore the amount of the effective ingredient may be in the range of the above range.
The kind of the food is not particularly limited. Examples of foods to which these substances can be added include meats, sausages, bread, chocolate, candies, snacks, cookies, pizza, instant noodles, other noodles, chewing gums, dairy products including ice cream, various soups, drinking water, tea, drinks, alcoholic beverages, and vitamin compounds, and all health functional foods in general are included.
The health drink composition of the present invention may contain various flavoring agents, natural carbohydrates, and the like as additional components, as in ordinary drinks. The natural carbohydrate is monosaccharide such as glucose and fructose, disaccharide such as maltose and sucrose, polysaccharide such as dextrin and cyclodextrin, and sugar alcohol such as xylitol, sorbitol and erythritol. As the sweetener, natural sweetener such as thaumatin and stevia extract, or synthetic sweetener such as saccharin and aspartame can be used. Generally, the proportion of natural carbohydrates is from about 0.01 to about 0.20g per 100ml of the composition of the invention, preferably from about 0.04 to about 0.10 g.
In addition to the above ingredients, the composition of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acids and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents for carbonated beverages, and the like. In addition, the composition of the present invention may contain pulp for producing natural fruit juice, fruit juice beverages and vegetable beverages. These ingredients may be used independently or in combination. Although the proportions of these additives are not critical, the amount is generally selected in the range of 0.01 to 0.20 parts by weight per 100 parts by weight of the composition of the invention.
As another aspect of the present invention, there is provided a method for treating lung cancer comprising the step of administering the pharmaceutical composition to a subject. In the present invention, the term "individual" refers to a subject to be treated for a disease, and more specifically refers to a mammal such as a human or non-human primate, rat (mouse), dog, cat, horse, and cow.
In the following, some preferred embodiments will be enumerated in order to facilitate understanding of the present invention. However, the following examples are given for the purpose of enabling the present invention to be more easily understood, and the contents of the present invention are not limited by the following examples.
[ examples ]
Example 1 apoptosis Effect of Lung cancer cell lines when treated with Honokiol, apigenin, curcumin alone or in combination
In order to confirm the effect of honokiol, apigenin and curcumin on apoptosis of lung cancer cell lines, human lung cancer cell line a549 (adenocarcinosic human avian basal epithelial cells) was treated with honokiol, apigenin and curcumin (Sigma-Aldrich) alone or in combination. The concentration of human lung cancer cell line treated with honokiol, apigenin and curcumin is determined by administration concentration curve (dose-concentration curve) as the concentration (IC) for inducing apoptosis to 20% when administered alone20Inhibition concentration 20), i.e., 10. mu.M, were subjected to separate and mixed treatments. After 24 hours of treatment, the apoptosis effect of lung cancer Cell lines was analyzed using CCK8assay (Cell Counting Kit-8, Dojindo Molecular Technologies, Inc).
As a result, as shown in fig. 1, the apoptosis effect was significantly increased in the mixed treatment compared to the case where honokiol, apigenin, or curcumin was treated alone. Specifically, the apoptosis induced by honokiol alone was 19%, that induced by apigenin alone was 22%, and that induced by curcumin alone was 20%, compared to the above results, the apoptosis induced by the mixture of honokiol and apigenin was 79%, and that induced by the mixture of honokiol and curcumin was 83%. It can thus be confirmed that the apoptotic effect caused by the mixture is not a purely additive effect (additive effect) but a synergistic effect (synergistic effect). Therefore, when a patient with lung cancer is treated, compared with the condition that honokiol, apigenin and curcumin are singly administered, the lung cancer treatment efficacy can be enhanced by the mixed administration of the honokiol and the apigenin or the mixed administration of the honokiol and the curcumin.
Example 2 ANT2 expression inhibitory Effect of Lung cancer cell lines when Honokiol, apigenin, curcumin alone or in combination
In order to confirm the inhibitory effect of ANT2 expression of honokiol, apigenin and curcumin, honokiol, apigenin and curcumin were treated individually or in combination with human lung cancer cell line A549. Then, in order to confirm that the expression of ANT2 mRNA of the lung cancer cell line was inhibited, qRT-PCR analysis was performed. More specifically, human lung cancer cell strain A549 was treated with 10 μ M honokiol, apigenin, and curcumin alone or in combination. After 24 hours, RNAs (TRIzol RNA Isolation Reagents, Thermo scientific) were extracted, and after cDNA was synthesized by reverse transcription (RT: reverse transcription) (PrimeScript 1st strand cDNA Synthesis Kit, Takara), real-time polymerase chain reaction (qPCR) was performed using SYBR (SYBR Premix Ex Taq, Takara). ANT2 (adenosine nucleotid translocase 2) as an analysis target is a gene known to play a very important role in the survival, proliferation and metastasis of cancer cells.
Analysis of the results of the qRT-PCR experiment utilized the following formula.
[fold change＝2-ΔΔCt,whereΔΔCt＝(Ct of gene of interest,treat ed-Ct of HK gene,treated)-(Ct of gene of interest,control-Ct of HK gene,control),Ct was the threshold cycle number and HK was the house-keeping gene.]
The base sequences of the primers used in qPCR are shown in Table 1 below.
[ TABLE 1 ]
As a result, as shown in fig. 2, it was confirmed that treatment of honokiol, apigenin, and curcumin alone or in combination resulted in a decrease in ANT2 expression. When compared with the case of treating honokiol, apigenin and curcumin separately, the expression inhibition effect of ANT2 is relatively higher when treated according to honokiol and apigenin or honokiol and curcumin.
Therefore, the curative effect similar to the cancer treatment effect generated by ANT2 small interfering RNA (shRNA) or siRNA (short interfering RNA) can be obtained by mixing and administering honokiol and apigenin or honokiol and curcumin. In addition, the reduction effect of ANT2 by treating honokiol, apigenin, and curcumin alone or in combination can reduce ANT2 using small interfering RNA, thereby solving the problems associated with delivery of siRNA or shRNA into the body (delivery system) in the treatment of cancer.
Example 3 ATP production reducing Effect of Lung cancer cell lines when Honokiol, apigenin, curcumin alone or in combination
The results in example 2 confirmed that the expression of ANT2 was effectively suppressed when human lung cancer cell lines were treated with honokiol, apigenin, and curcumin alone or in combination. Therefore, due to the crucial role of ANT2 function in ATP production of cancer cells, it is expected that treatment of honokiol, apigenin, curcumin alone or in combination will lead to a decrease in ATP production of cancer cells.
Therefore, in order to confirm the effect of suppressing ANT2 expression to reduce ATP production in lung cancer cell lines, human lung cancer cell line a549 was treated with honokiol, apigenin, and curcumin alone or in combination. Then, using CellTiter-GloTMLuminescent Cell visual assay kits (Promega) were analyzed.
As a result, as shown in fig. 3, when honokiol, apigenin, and curcumin were treated alone or in combination, it was confirmed that ATP production of human lung cancer cell lines was effectively reduced. In particular, when honokiol + apigenin, honokiol + curcumin are mixed and administered, the ATP production reducing effect is relatively enhanced as compared with the case of single administration.
It is thus judged that a decrease in ATP, which is an essential energy source for survival, inhibits the growth of cancer cells and ultimately induces apoptosis of cancer cells, and therefore, administration of honokiol, apigenin, and curcumin alone or in combination inhibits the expression of ANT2, thereby inducing a decrease in ATP production. Therefore, the cancer treatment effect can be improved by the inhibitory effect on ATP production in cancer cells.
Example 4 HK2 expression inhibitory Effect of Lung cancer cell lines when Honokiol, apigenin and curcumin were treated alone or in combination
In order to confirm whether inhibition of ANT2 expression by treatment of honokiol, apigenin, and curcumin alone or in combination regulated the expression of hexokinase 2(HK2), an enzyme that interferes with glycolysis, honokiol, apigenin, and curcumin alone or in combination were treated with human lung cancer cell line a549 as a target. Then, analysis was performed using qRT-PCR.
Analysis of the results of the qRT-PCR experiment utilized the following formula.
[fold change＝2-ΔΔCt,whereΔΔCt＝(Ct of gene of interest,treated-Ct of HK gene,treated)-(Ct of gene of interest,control-Ct of HK gene,control),Ct was the threshold cycle number and HK was the house-keeping gene.]
The base sequences of the primers used in qPCR are shown in Table 2 below.
[ TABLE 2 ]
As a result, as shown in fig. 4, it was confirmed that the expression of HK2 was significantly reduced when honokiol, apigenin, and curcumin were treated alone or in combination, and that the inhibitory effect on the expression of HK2 was relatively higher when honokiol + apigenin or honokiol + curcumin were treated in combination, as compared with the case where honokiol, apigenin, and curcumin were treated alone.
The HK2 enzyme was expressed in small amounts in normal cells, but the expression level of HK2 was very high in most cancer cells. In addition, the HK2 enzyme not only phosphorylates glucose, but also binds to mitochondrial VDAC protein to hinder the apoptotic mechanism (Apoptosis). This can enhance the viability of cancer cells. Therefore, treatment of honokiol + apigenin mixture or honokiol + curcumin mixture resulted in significant inhibition of HK2 expression, which could enhance its therapeutic effect during cancer treatment targeting HK 2.
Example 5 glycolysis Rate reducing Effect of Lung cancer cell lines when treated with Honokiol, apigenin, curcumin alone or in combination
From the results of example 4, it was confirmed that treatment of honokiol, apigenin, and curcumin alone or in combination had the effect of inhibiting the expression of HK 2. Therefore, in order to confirm whether glycolysis rate of lung cancer cell line could be reduced by treating honokiol, apigenin, and curcumin alone or in combination, honokiol, apigenin, and curcumin alone or in combination were treated with human lung cancer cell line a549 as a target. Then, analysis was performed using the Lactate Assay Kit (Biovision).
As a result, as shown in fig. 5, it was confirmed that treatment of honokiol, apigenin, and curcumin alone or in combination resulted in a reduction in glycolysis in human lung cancer cell lines. In particular, when honokiol + apigenin or honokiol + curcumin are mixed and administered, the glycolysis reduction effect is enhanced as compared with the case where honokiol, apigenin, and curcumin are administered alone.
In addition, since the oxidative phosphorylation pathway of mitochondria is defective in the glucose metabolism of cancer cells, the efficiency of ATP production by glycolysis is lower than that of oxidative phosphorylation, assuming that ATP is produced mainly only by glycolysis. Therefore, cancer cells must perform the glycolytic process with very high standards in order to produce sufficient ATP. Therefore, it is estimated that metabolic inhibition by inhibition of cancer metabolism-related enzymes is highly valuable as a target for cancer treatment. It is expected that the glycolysis reduction effect by the mixed treatment of honokiol + apigenin or honokiol + curcumin will be effectively applied to the treatment of lung cancer.
Example 6 inhibition of PD-L1 expression in Lung cancer cell lines by treatment of Honokiol, apigenin, and curcumin alone or in combination
In order to confirm the effect of inhibiting the expression of PD-L1 in human lung cancer cell lines by treating honokiol, apigenin and curcumin alone or in combination, honokiol, apigenin and curcumin were treated alone or in combination with human lung cancer cell lines A549 and H2228. Then, the degree of expression in the PD-L1 protein standard was analyzed by Western blotting (Western-blot) (PD-L1 antibody; cell-signalling, action antibody: santa cruze).
As shown in FIG. 6a, the lung cancer cell line A549 used in examples 1 to 5 had very low expression of PD-L1 (PD-L1)low) Therefore, it was determined that it was difficult to confirm whether or not expression of PD-L1 was reduced by honokiol, apigenin, and curcumin, and therefore, a lung cancer cell line H2228 (PD-L1) having high expression of PD-L1 was usedhigh)。
As a result, as shown in fig. 6b, it was observed that the expression of PD-L1 was effectively suppressed when honokiol, apigenin, and curcumin were treated alone or in combination. In particular, a reduction in PD-L1 expression was weakly observed when honokiol, apigenin, or curcumin were treated alone, but a very strong inhibition of PD-L1 expression was observed when honokiol + apigenin or honokiol + curcumin were mixed.
If a T cell having an effect of killing cancer cells binds to PD-L1, which is a protein present on the surface of cancer cells, the T cell cannot attack the cancer cells, and the effect of killing cancer cells is lost. The immunoanti-cancer agent prevents the antibody from binding to the PD-L1 protein, thereby allowing the T cell to actively attack the cancer cell. The anticancer agent enhances the immune system of human body, so there is no side effect or tolerance caused by the existing anticancer agent. Therefore, as one of the immune evasion strategies of cancer cells, the function of tumor-specific T lymphocytes can be inhibited by the expression regulation of PD-L1 and by utilizing the change of immune checkpoint (immune checkpoint) functions. That is, this suppressive immune checkpoint is activated within the tumor cell, so that attack by tumor-specific T-lymphocytes can be evaded. It has recently been known that the function of PD-L1 is inhibited by a monoclonal antibody, and the activity and effect of tumor-specific T-lymphocytes are enhanced, whereby an antitumor effect can be obtained. The treatment method targeting PD-L1 achieved a very effective clinical effect on a variety of cancer cells that has never been achieved before.
From this, it was confirmed that the inhibition of ANT2 or HK2 expression by the mixed administration of honokiol + apigenin or honokiol + curcumin not only directly induces apoptosis, but also overcomes the immune evasion mechanism of lung cancer cells by PD-L1 inhibition. Thus, the effect of cancer treatment will be enhanced in the end.
The above description of the present invention is exemplary, and it should be understood that a person having ordinary skill in the art to which the present invention pertains can easily change the present invention to other specific forms without changing the technical idea or essential characteristics of the present invention. The embodiments listed above are therefore exemplary in all respects and do not limit the scope of the invention.
Industrial applicability of the invention
The composition according to the present invention can induce apoptosis of cancer cells ascending each other when treated with a mixture or in combination, as compared with the case where apigenin, curcumin or honokiol is treated alone. The pharmaceutical composition for preventing or treating lung cancer, which contains either apigenin or curcumin and a honokiol mixture as active ingredients according to the present invention, has an improved anticancer effect, can induce apoptosis of cancer cells, and exhibits a very excellent cancer treatment enhancing effect. Therefore, the composition according to the present invention can be effectively applied to the prevention, amelioration or treatment of lung cancer.
<110> Bai ao Yi Fu Bing Ltd
<120> pharmaceutical composition for preventing or treating lung cancer comprising apigenin, curcumin and honokiol as active ingredients
<130> DFP19KR3534
<150> KR 10-2016-0111383
<151> 2016-08-31
<160> 8
<170> KoPatentIn 3.0
<210> 1
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> ANT2_forward primer
<400> 1
<210> 2
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> ANT2_reverse primer
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gtgtcaaatg gataggaag 19
<210> 3
<211> 20
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<213> Artificial Sequence
<220>
<223> GAPDH_forward primer
<400> 3
<210> 4
<211> 20
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<213> Artificial Sequence
<220>
<223> GAPDH_reverse primer
<400> 4
<210> 5
<211> 20
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<213> Artificial Sequence
<220>
<223> HK2_forward primer
<400> 5
<210> 6
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<213> Artificial Sequence
<220>
<223> HK2_reverse primer
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gccaggtcct tcactgtctc 20
Claims (2)
1. Use of a composition in the manufacture of a medicament for the prevention or treatment of lung cancer,
the composition is prepared from apigenin or curcumin; and honokiol as an active ingredient, and the composition reduces the expression of ANT2 gene.
2. Use of the composition of claim 1 in the manufacture of a medicament for the prevention or treatment of lung cancer, wherein the composition reduces the expression of HK2 or PD-L1 protein.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
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| KR10-2016-0111383 | 2016-08-31 | ||
| KR1020160111383A KR101803000B1 (en) | 2016-08-31 | 2016-08-31 | Pharmaceutical compositions for preventing or treating lung cancer comprising apigenin, curcumin, and honokiol as active ingredients |
| PCT/KR2017/009241 WO2018043989A1 (en) | 2016-08-31 | 2017-08-24 | Pharmaceutical composition comprising apigenin, curcumin, and honokiol as active ingredients for preventing or treating lung cancer |
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| CN109640978A CN109640978A (en) | 2019-04-16 |
| CN109640978B true CN109640978B (en) | 2021-08-27 |
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| Country | Link |
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| KR (1) | KR101803000B1 (en) |
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| CN113952351B (en) * | 2021-09-08 | 2023-02-17 | 隋新兵 | Pharmaceutical composition for treating cancer and preparation method and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1692901A (en) * | 2004-05-08 | 2005-11-09 | 陈菲 | Application of magnolol for preparing medicine compositions to treat and prevent tumor |
| US20120171228A1 (en) * | 2011-01-05 | 2012-07-05 | Econugenics, Inc. | Anti-cancer regimen |
| US8377918B2 (en) * | 2005-01-31 | 2013-02-19 | ACC Therapeutics Inc | Apigenin for chemoprevention, and chemotherapy combined with therapeutic reagents |
| US20130129809A1 (en) * | 2011-05-19 | 2013-05-23 | Glax L.L.C. | Compositions and methods for treating and preventing cancer by targeting and inhibiting cancer stem cells |
| WO2014192024A1 (en) * | 2013-05-20 | 2014-12-04 | Sree Chitra Tirunal Institute For Medical Sciences And Technology | Development of soluble albuminated curcumin for application in cancer therapy |
| US20160146783A1 (en) * | 2012-04-05 | 2016-05-26 | The Regents Of The University Of California | Compositions and methods for treating cancer and diseases and conditions responsive to cell growth inhibition |
-
2016
- 2016-08-31 KR KR1020160111383A patent/KR101803000B1/en active IP Right Grant
-
2017
- 2017-08-24 WO PCT/KR2017/009241 patent/WO2018043989A1/en active Application Filing
- 2017-08-24 CN CN201780052022.4A patent/CN109640978B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1692901A (en) * | 2004-05-08 | 2005-11-09 | 陈菲 | Application of magnolol for preparing medicine compositions to treat and prevent tumor |
| US8377918B2 (en) * | 2005-01-31 | 2013-02-19 | ACC Therapeutics Inc | Apigenin for chemoprevention, and chemotherapy combined with therapeutic reagents |
| US20120171228A1 (en) * | 2011-01-05 | 2012-07-05 | Econugenics, Inc. | Anti-cancer regimen |
| US20130129809A1 (en) * | 2011-05-19 | 2013-05-23 | Glax L.L.C. | Compositions and methods for treating and preventing cancer by targeting and inhibiting cancer stem cells |
| US20160146783A1 (en) * | 2012-04-05 | 2016-05-26 | The Regents Of The University Of California | Compositions and methods for treating cancer and diseases and conditions responsive to cell growth inhibition |
| WO2014192024A1 (en) * | 2013-05-20 | 2014-12-04 | Sree Chitra Tirunal Institute For Medical Sciences And Technology | Development of soluble albuminated curcumin for application in cancer therapy |
Non-Patent Citations (5)
| Title |
|---|
| Construction and application of a lung cancer stem cell model: antitumor drug screening and molecular mechanism of the inhibitory effects of sanguinarine;Jia Yang;《Tumor Biol》;20160802;第1-13页 * |
| 几种中药有效成分联合应用对小鼠肉瘤细胞株S180的作用;肖春芬;《中国药房》;20070330;第18卷(第9期);第653-655页 * |
| 和厚朴酚对人肺小细胞癌N446细胞增殖与凋亡的影响;黄清松;《长春中医药大学学报》;20131231;第29卷(第6期);第967-969页 * |
| 姜黄素( Curcumin)能够升高肺癌细胞对抗肿瘤药物的敏感性;马德宾;《科学技术与工程》;20150828;第15卷(第24期);第23-28页 * |
| 芹菜素对肺癌细胞的抗肿瘤作用以及增敏作用分析;黄再超;《World Latest Medicine Information》;20150929;第15卷(第78期);第118、126页 * |
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| KR101803000B1 (en) | 2017-11-30 |
| WO2018043989A1 (en) | 2018-03-08 |
| CN109640978A (en) | 2019-04-16 |
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