TW201119663A - Composition of Chinese medicine recipes for cancer therapy. - Google Patents

Abstract

The present invention relates to a Chinese medicine composition used to treat cancer, which comprises a combination of catechin, curcumin and phyllanthus emblica extract. The Chinese medicine composition according to the present invention composed of catechin, curcumin and phyllanthus emblica extract can effectively inhibit the growth of cancer cell, enhance the apoptosis of cancer cell, and repress the growth and development of tumor. The present invention further proves that the Chinese medicine composition can inhibit the spread of cancer cell by increasing cell adhesion ability, enhance cell differentiation and cause tumor suppression regulation mediated by contact inhibition.

Description

201119663 VI. Description of the Invention: [Technical Field] The present invention relates to a composition comprising catechin, curcumin and emblica extract, which can be used for treating cancer, more particularly for treatment display A cancer having high epidermal growth factor receptor (EGFR) expression, such as oral cancer, and for treating or preventing tumor metastasis. [Prior Art] Cancer formation is associated with genetic factors, inflammatory responses, oxidative stress, and protein kinase activation (Franco, R. et al., deer 266(1): 6-11, 2008;

Alberti, C., Eur Rev Med Pharmacol Sci. 12(3): 167-75, 2008) ° ^ During cancer formation, cancer cells accumulate self-mutation so that the tumor cell population in the patient is heterogeneous; In addition, each patient's constitution <gene background> is different', so it will affect the success rate of treatment and the survival rate of patients (Kunz, Μ" 0/7T5(1): 9-14, 2008). The inflammatory response of the surrounding tiny environment will help the proliferation and survival of tumor cells, promote blood blasts and tumor metastasis, destroy the acquired immune response and change the susceptibility of cancer patients to chemotherapy drugs (Mantovani, A· et al., 454 (7203)· · 436-44, 2008.) Epidemiological observations show that oxidative stress is one of the main factors in the process of cancerization. Under adverse conditions, cells that adapt and proliferate are screened out and free radicals are modified. Body information is relevant (s ·, X X 60(7): 441-7, 2008). Protein stimulating is a group of enzymes that importantly control the growth and differentiation of cells 201119663. When they are not functioning properly, they will participate in certain Tumor development. For example, literature Guide noted oral cancer, bladder cancer, lung cancer, pancreatic cancer, and Jie Tang stimulated EGFR protein expression transition situation (Chang, Jw, et al., Cancer6l (3): 328-39, 2008; Konkimalla, Expert Rev

Anticancer Ther 7(3): 317-29, 2007 ; Wang, X. et al., Clin Cancer Resl 3(3): 953-7, 2007; Jimeno, A. et al., 68(8): 2841-9, 2008 ). Therefore, if these carcinogenic factors can be effectively controlled, it will be effective in assisting the clinical treatment of cancer. Natural plant extracts with functional regulation, because they can be combined with daily diet and maintenance, have been extensively developed into cancer-assisted therapeutic products, such as catechins from camellia leaves and curcumin from underground stems. These natural plants have been used as health foods in Asian countries for centuries. They all have antioxidant, anti-inflammatory, anti-cancer and the ability to stop cancer cells from advancing the cell cycle and inducing apoptosis (Chen, D. et al., et al. / cf. / 23(4): 487-96 , 2008; Reuter, S. et al., 10,000/oc/zem P/zarmaco/, 2008). According to cell cycle analysis, catechins stop cancer cells in sub-Gl or G1 phase; curcumin stops cancer cells in G2/M phase (Khafif, A. et al., Garcz. wogewe also 19(3) : 419-24, 1998 ; Harakeh, S. et al., 25(1): 30-9, 2008; Liu, E. et al., J Neurooncol 85(3): 263-70, 2007). In addition, the extract of Phyllanthus emblica L. is rich in vitamin C, minerals, amino acids and various phenolic compounds (Fujii, T. et al., J and /119(1): 53-7, 2008), thus Excellent antioxidant capacity' is an important formula for many skin care today, while Yu 5 201119663

Ganzi also has anti-inflammatory and anti-inflammatory side effects (Haque R et al., Seto Tbx such as / 20 (12): 643-50, 2001; Asmawi, MZ et al. 45 (6): 581-4, 1993),. Because there are many lines of message transmission that control the formation of cancer, combining multiple natural plant extracts will provide more effective anti-cancer effects (Sagar, s m., d. Yanee and RK Wong, CWr Ο /, 2006. 13(1): 14·26). In cell model and animal experimental models, a number of combinations of natural plant extracts have been proposed. For example, it has been reported that catechin and the EGFR protein kinase inhibitor erlotimb synergistically inhibit the growth of head and neck cancer cells (Zhang, et al., j 123 (5): 1005-14, 2008). The catechins and antioxidants such as resveratrol and γ-saltene can synergistically enhance the performance of Nq〇i enzymes in metabolic chemotherapy drugs, in order to improve the therapeutic effect of chemotherapy drugs (Hsieh, TC and JM Wu, /咐J Owe ?/. 33(4): 851_9, 2008). Low doses of curcumin have been reported to be combined with antioxidants such as vitamin C, N-acetyl-cysteine and reduced glutathione to enhance the anti-cancer ability of curcumin (Chen, j. Wait,

60(1): 57-61, 2005). Recent studies have further indicated that the combination of curcumin and catechin has the synergistic effect of inhibiting the growth of oral cancer and breast cancer cells (Khaflf, A et al., supra; Somers-Edgar, TJ et al, MJ Cancer 122 (9) : 1966-71, 2008). Natural plant extracts are also used extensively in clinical research. For example, urinary tract can be used to treat breast cancer or prevent breast cancer recurrence in stage I and II, and has anti-cancer effects on colorectal cancer, pancreatic cancer, stomach cancer, ovarian cancer and lung cancer (Moyers, 201119663 SB and NB fire wmar, iVwirAv 62(5): 204-11, 2004). Curcumin has anticancer effects on colorectal cancer, pancreatic cancer, cervical cancer, head and neck or oral cancer (Hatcher, H. et al., CW/Mo/Z% 5W 65(11): 1631-52, 2008) . Traditional chemotherapeutic agents have many negative effects. If you can combine traditional chemotherapeutic treatments with many different and mild natural plant extracts, it is believed that synergistic effects will increase the anticancer effect and reduce the toxicity of the drug to normal tissues ( Patel, Β.Β. et al., /m JC Dirty 122(2): ρ. 267-73, 2008 ; Ozcan, Α. • et al., such as Coronation (7) / 79 (8): 461-5, 2005). Therefore, by combining different agents, it is possible to overcome the development of drug resistance and enhance the therapeutic effect (Sagar, SM, D.

Yance and R.K. Wong, as mentioned above; Sathomsumetee, S. et al., Ca, r 110(1)·13-24, 2007). In addition, a combination of therapeutic agents such as anti-angiogenic factor monoclonal antibody bevadzumab combined with clinical standard therapy, clinically applied to lung cancer, pancreatic cancer, breast cancer and metastatic renal cell carcinoma patients have good results (de Gramont, A. and E. Van Cutsem, 〇_/〇 at Suppi &amp; p • 46-56, 2005). Yu Ganzi is one of the ancient fruits used by Indians. Previous studies have indicated that NB1RGB human skin fibroblast strains indicate that Phyllanthus emblica has therapeutic or cosmetic efficacy by controlling the control of prion protein (Fujii, T. et al., supra), and in the animal experiments of the present invention. It can be seen that the skin color of nude mice with healthy yam is indeed smoother than other groups (see Figure 6B); it has been proven to enhance memory in animal experiments (Vasudevan, M. and Μ Parle, Cai Yang 91(1): 46_54, 2〇〇7) and effective treatment of hyperlipidemia and prevention 201119663 HJ. ^^JNutr Sci Vitaminol (Tokyo) 51(6). p. 413-8, 2005). However, there are few reports on the mechanism of anti-cancer effects of Phyllanthus emblica. For example, in animal experiments, it has been found that Phyllanthus emblica can resist inflammation (AsmaWi, MZ et al., as mentioned above) and reduce the immunity caused by chemotherapy drugs ( Haque'R. et al., supra). Phyllanthus emblica L. can reduce the loss of renal function in rats due to oxidative stress due to age (Y〇k〇zawa, τ. et al., j C/zem 55(19): ρ· 7744-52, 2007). Therefore, according to these concepts, the present invention firstly combines catechin, curcumin and emblica extract for the treatment and/or prevention of cancer, and proves that catechin is used in combination with animal experiments of cell strains and xenograft tumors. The combination treatment of curcumin and Phyllanthus emblica extract can produce a synergistic effect, and the present invention can be accomplished by reforming the critical pathways in cancer cells and reducing the oncogenic activity of regulating them to achieve the effects of treating and preventing cancer and tumor metastasis. SUMMARY OF THE INVENTION In one aspect, the present invention provides a traditional Chinese medicine composition for treating an individual suffering from cancer, comprising 20% to 60% catechin, 20% to 60% curcumin and 20% to 60% 5% of the nectar extract, as well as pharmaceutically acceptable carriers, diluents or excipients. The proportion of ingredients can be adjusted according to different cancer types. In one embodiment, the traditional Chinese medicine composition comprises 20% to 60% catechin, 2 〇〇/0 to 40% curcumin and 20% to 40% cumin extract. In another embodiment, the Chinese medicinal composition comprises 50% catechin, 25% curcumin, and 25% 201119663 cumin extract. In one embodiment of the invention, the tumor is a cancer having a high EGFR expression. In another embodiment of the invention, the tumor is oral cancer. In another aspect, the present invention provides a pharmaceutical composition for promoting apoptosis of cancer cells comprising 20% to 60% catechin, 20% to 60% curcumin and 20% to 60% Ganzi extract, as well as pharmaceutically acceptable carriers, diluents or excipients. The proportion of ingredients can be adjusted according to different cancer types. In one embodiment, the traditional Chinese medicine composition comprises 20% to 60% catechin, 20% to 40% curcumin and 20% to 40% geranium extract. In another embodiment, the traditional Chinese medicine composition comprises 50% catechin, 25% curcumin, and 25% Phyllanthus extract. In a specific aspect of the invention, the cancer cell is a cancer cell having a high EGFR expression. In another embodiment of the invention, the cancer cell is an oral cancer cell. In yet another aspect, the present invention provides a pharmaceutical composition for inhibiting tumor metastasis comprising 20% to 60% catechin, 20% to 60% curcumin and 20% to 60% nectar An extract, as well as a pharmaceutically acceptable carrier, diluent or excipient. The proportion of ingredients can be adjusted according to different cancer types. In one embodiment, the composition of the traditional Chinese medicine comprises 20% to 60% catechin, 2% to 40% curcumin and 20% to 40% nectar extract. In another embodiment, the traditional Chinese medicine composition comprises 50% catechin, 25% curcumin and 25% yam extract. 201119663. In the specific aspect of the present invention, the effect of the pharmaceutical composition for inhibiting tumor metastasis is to inhibit the spread of cancer cells and promote the differentiation of cancer cells by regulating cell attachment ability, thereby causing contact inhibition between cancer cells. [Embodiment] The specific terms used in the present invention have their original meanings, and some specific terms used as follows are provided to those skilled in the art to further understand the contents of the present invention. Unless otherwise defined, the scientific and technical vocabulary involved in this riding is the same as the vocabulary used in general skills. As used herein, "cancer," means any cell disorder in which the tissue proliferation of the tissue is corrected = rapid tissue. Cancer includes, but is not limited to, oral cancer, facial cancer, breast cancer, brain cancer (glioma). , lung cancer, prostate cancer 'colorectal cancer, thymus cancer, kidney cancer, kidney coal cancer, liver cancer, multiple neurofibromatosis and leukemia. Cancer also includes solid cancer (such as sarcoma or cancer), or affect the cancerous growth of the hematopoietic system (eg lymphoma or leukemia). EGFR-expressing cancer, meaning a cancer cell whose isolated cell line is known to have high EGFR-expression, including, for example, bladder cancer, cancer, colorectal cancer, breast cancer, lung cancer ( For example, small cell lung cancer), gastric cancer, sputum cancer, prostate cancer, head and _, kidney cancer, gallbladder cancer and oral cancer. 匕 For this test x Mingzhimu #, "To treat individuals suffering from cancer, you should understand the intention 'Although compared with the traditional Chinese medicine containing catechin, curcumin and emblica extract without the present invention, the person who has been bribed when the reduction is transferred has reduced or weakened the symptoms or conditions of New Zealand. The treated cancer condition can be caused by, for example, inhibition of growth of cancer cells, increase in cancer cell growth and/or reduction in tumor mass, rate of increase in pre-inflammation, reduction in tumor morphology, and reduction in the facial side. The severity is reduced and the second is confirmed as compared to the increase in the life of the individual who has not received the treatment. In a broad sense - when the field receives the desired clinical response, it should be considered that successful treatment has occurred. For example, successful treatment can be defined by obtaining, for example, 1% or more (i.e., sputum, 30%, 40%) tumor growth inhibition. An individual is a mammal, preferably a human, but may also be an animal that requires veterinary treatment, such as a pet (eg, a dog, a june, a farm animal (eg, a cow, a lamb, a pig, a horse, etc.) and an experiment. Animals (eg rats, mice, guinea pigs, etc.). 1. In the traditional Chinese medicine composition containing the catechin, curcumin and emblica extract of the present invention, the exact individual individual drug administered to the individual will depend on the type and severity of the disease or condition, and the individual Characteristics such as general health, age, sex, weight and tolerance to the drug. It also depends on the extent, severity and type of cancer. Applicants who are skilled in the art can determine the appropriate dose based on these and other factors. The pharmaceutical composition of the present invention can be administered by any suitable route, including, for example, oral administration in capsules, suspensions, or parenteral administration. Parenteral administration can include, for example, total miscellaneous, e.g., by intramuscular, intravenous, subcutaneous, or intraperitoneal injection. The compound can also be administered orally (e.g., by eating), topically, via inhalation (e.g., intrabronchial, intranasal &apos; inhalation, 201119663, intranasal drip) or suture material rhyme, which is desirable for the type of red axis. It is better to use medicinal or parenteral drugs for administration.

The preparation method of the W-complex of the present invention will vary according to the selected administration route, such as a solution, a powder, a face capsule, and the like. Suitable pharmaceutical carriers may contain anti-narrative components that do not interact with the compound. Fresh medicine modulation techniques can be described, for example, in Remington's Medical Sciences, Merck Publishing Company, Eston, and others. Examples of solid carriers suitable for oral administration include, for example, lactose, white clay, sucrose, cyclodextrin, talc, gelatin, _, fruit ginseng, gum arabic, magnesium stearate, stearic acid, and cellulose; Examples include, for example, (iv), peanut oil, eucalyptus oil, phospholipids, fatty acids, fatty acid amines, polyethylene oxide water. The medical laboratory of the non-intestinal county includes, for example, extinguishing water, physiological saline, bacteriostatic saline (food containing % mg/ml benzyl alcohol), Gu Yan _ food, money , Ringer's lactate and the like. Methods for encapsulating hybrids (e.g., encapsulated in a hard or oligosaccharide envelope) are known in the art (see, for example, Baker et al., "Controlled Release of Bioactive Agents", about round Wiley and his son published, Cong). It is required that the Chinese music group containing the catechin, curcumin and emblica extract of the present invention can be combined with other anti-paste agents, such as purple transgenic, Changchun new test, etoposide: subfamily , actinomycin D, mitomycin C, bleomycin, periwinkle test, cis-ammonium J white, monoclonal antibody, etc., to reduce the clinical use of these known anti-cancers, In turn, the side effects produced by the anticancer agents are reduced or alleviated, and the drug resistance may occur in the (4) like individuals. The other features and advantages of the present invention are further exemplified and described in the following examples. EXAMPLES Example 1: Oxidation of catechins, curcumin and ambroin in plasma and changes in antioxidant capacity First, the individual components (EGCG), curcumin (Cur) and the individual components of the traditional Chinese medicine composition of the present invention were tested. The effect of Yugan and two other antioxidant green tea extracts (GTE) and Rhodiola rosea extract (RR) on plasma antioxidant b-force. When the plasma is in contact with air, the blood will begin to be air. Oxygen is oxidized. Therefore, the longer the plasma is exposed to oxygen, the more the blood is oxidized. In contrast, the plasma oxidative stress (Gxidativestress) is more serious. When the plasma oxidative pressure is increased, the blood oxidative metabolism The concentration of the substance will increase and the total antioxidant capacity of the plasma will decrease. If an antioxidant is added to the plasma, the antioxidant will lower the oxidative pressure of the plasma and increase the total antioxidant capacity of the plasma. Therefore, the anti-oxidation is added to the plasma. After the oxide, the lower the agronomic oxidative metabolite of plasma A or the higher the total antioxidant capacity, the higher the antioxidant capacity of the antioxidant. Plasma nitrogen oxides (Pla) The sma NOx) concentration detection method was modified from the literature published by Moshag et al. in 1995 (M〇shage, H et al., l l l S] 13 201119663 C/^41 (6 Pt 1): 892-6, 1995) The extracts (1 mg/ml) were mixed with plasma at a ratio of 5:100 and then allowed to stand at room temperature for 2 hours. After that, an equal volume of Griess reagent was added and allowed to stand at room temperature for 15 minutes. The absorbance at 540 nm wavelength is expressed as the equivalent number of NaN〇2. The plasma lipid metabolite (Plasma TBARS) concentration detection method is modified from the literature published by kawahkawa et al. in 1997 (Ohkawa, H., N). Ohishi and K. Yagi, 95(2): 351-8, 1979). Each extract (1 mg/mi) was mixed with plasma at a ratio of 5:100 and then allowed to stand at room temperature for 2 hours.丨00 ui plasma was sequentially added with 100 ul of physiological saline, 200 ul of TCA solution (2 g of tri-glycolic acid dissolved in 2 ml of pure water) and 50 ul of TBA solution (20 mg of thiobarbituric acid ( Thiobarbituric acid) dissolved in 6 ml of 50% acetic acid solution), mix well and then bathe at 85_C for 1.5 hours. Add 60 ul of !1-butanol, mix and centrifuge at 3500g for 1 minute, remove the supernatant. The liquid reads the absorbance of 52 ,, and the data is expressed as absorbance. The method for detecting the concentration of blood peroxides is modified from the literature published by Sodergren et al. in 1998 (Sodergren, E. et al. J, she added 37 (3): 137-46, 1998). Each extract (1 mg/mi) was mixed with plasma at a ratio of 5:1 Torr, and allowed to stand at room temperature for 2 hours. 100 ul of plasma was added to 9 〇〇U1 of F0X2 reagent, mixed and allowed to stand at room temperature for 7 minutes, then centrifuged at 35 〇〇g for 10 minutes, and the supernatant was taken to read its absorbance at 560 nm. The data is expressed as the number of equivalents of hydrogen peroxide. For the Ferric reducing/antioxidant power (FRAP) test, each extract (1 mg/ml) 201119663 was mixed with plasma at a ratio of 5:100, and then allowed to stand at room temperature for 2 hours. Then, the plasma was diluted 10 times with pure water, and 20 μl of the diluted plasma was added to the FRAP reagent of 〇.5 mi, and after standing at room temperature for 4 minutes, the absorbance at 620 nm was read, and the obtained data was sulfuric acid. The number of equivalents of ferrous iron is expressed. The prooxidant effect detection method for each extract was modified from the literature published by Tsai et al. in 2003 (Tsai et al., 51(1): ρ. 58-62, 2003). Each extract (0.1 mg/ml) 100 u of pure water 750 ul and 1 ul of luster φ (lucigenin) were mixed in a chemiluminometer (CLA-FS1, Tohoku Electronic)

Industrial Co. Ltd., 6-6-6 Shirakashidai, Riiu Town, Miyagi, Japan) was allowed to stand for 20 seconds, after which the detection of light was detected until 6 seconds later. The resulting data is expressed in terms of the number of photons (CL counts). The results of Table 1 show that different extracts, after being dissolved in plasma, exhibit different anti-emulsification capabilities and oxidative metabolites (such as NOx, TBARS and peroxide) scavenging ability. After plasma was added to five different antioxidants, the plasma Φ and the total antioxidant capacity (FRAP) of the plasma were varied to varying degrees. In terms of the test items of plasma oxidative metabolite Nox, the antioxidant capacity of each extract was EGCG &gt; AMLA &gt; GTE &gt; Cur &gt; RR. In terms of the TBARS test for plasma oxidative metabolites, the antioxidant capacity of each extract was: GTE &gt; AMLA &gt; RR = Cur &gt; EGCG. In terms of plasma peroxide testing items, the antioxidant capacity of each extract was: EGCG &gt; GTE = AMLA = Cur&gt;RR. In terms of plasma oxidative capacity (FRAP) test items, the antioxidant capacity of each extract was EGCG>GTE>RR&gt;AMLA = 15 201119663 cur. In terms of oxidative capacity, the ability is: gte>egcg &gt; RR>AMLA &gt; Cur 〇 Table - Analysis of the antioxidant capacity and oxide scavenging ability of various extracts after being dissolved in plasma

TBARS Peroxidation FRAP to promote oxidative capacity (delivery fraction (〇D540xl〇〇) (nM), photon photon number) 662666±5811 461157±9985 25883±1008 100245±4469 953士53 17 18 17 12 15 209±10 30.6 ±0.4 184±3 233±14 30.0±0 191±3 224±23 30.6±1 132±2 229±25 36.0±0.4 178±3 229±31 30.6±1 132±2 If the antioxidant capacity and each oxide The ability to clear up is ranked by the score, that is, five points in the first place, four points in the second place, and so on. From the above-mentioned low ranking results, EGCG is obtained in the comprehensive evaluation. 18 points, GTE and AMLA scored 17 points, Cur scored 15 points, and RR scored 12 points (see Table 1). Of course, the higher the score obtained by the comprehensive evaluation, the stronger the antioxidant capacity of this antioxidant. Therefore, it can be seen from the above that the order of oxidation resistance of the above five extracts is: EGCG &gt; GTE = AMLA &gt; Cur &gt; RR. In the above five extracts, EGCG and GTE are the same kind of extracts extracted from green tea 'only GTE is green tea extract' containing a variety of mixed tea polyphenols or catechins; and EGCG is Pure 201119663 chemical type of single tea polyphenols. Therefore, if the composition is to be developed to develop an antioxidant compound with high antioxidant capacity, it is expected that the combination of EGCG+AMLA or the combination of EGCG+AMLA+Cm· can obtain the best antioxidant capacity and promote oxidative capacity. The lowest (toxic) composition. Example 2: Containing catechin, curcumin and Phyllanthus emblica L. to synergistically inhibit cancer cell growth and community formation _ The cell strains SAT, KON, BIC and SEG used in this test are oral squamous cells. Cancer cell line. The SAT cell line used rpmi_164 〇 medium (purchased from GIBCO); KON, BIC and SEG all used DMEM (purchased from GIBCO). All cell lines were supplemented with 1% fetal calf serum (purchased from GIBCO) in the medium '1% antibiotics (penicillin and streptomycin, purchased from GIBC〇), 37 C cell incubator, 5% carbon dioxide environment Cultivate. Replace the medium once every two to three days.

• To understand the optimal concentration of natural plant extracts, use SYBR

The Green I Acacia Dyeing Reagent was purchased from the method to analyze the effect of different substances/lengths (from 0 to 1 gg #m) on the growth of oral squamous cell carcinoma cell lines. The oral cancer cell line was maintained in 1 〇/〇 fetal calf serum before the drug-treated cell line, and then 5 to 3 cancer cells per well were seeded in 96-well cell culture. Inside the plate, cultivate until overnight. Add 〇~i〇〇/m catechin, tomb flavonoids and/or amlaose the next day. In this experiment, 201119663 was treated with 0.02% DMSO as an experimental control group of catechin and Phyllanthus emblica. 96 hours after dosing, the medium in the culture plate was blotted, 50//1 0.2% SDS was added to each well, and then 37 was placed. (: 3 min in the cell culture incubator. After that, add SYBRGreen I 'isoy 丨 (1:1_dilution) to each well and mix it evenly with a fluorescence analyzer at an excitation wavelength of 485 Cong; absorption wavelength is 530 The absorbance was measured under the condition of nm. This experiment was repeated three times. Referring to the results shown in Fig. 1, after 4 days of dosing, catechins can be observed against SAT, KON, BIC and SEG cells, inhibiting 25% of cells. The effective active concentrations for growth were 10//M, 30/zM, 45//M, and 40//M, respectively (Fig. 1A); curcumin inhibited SAT, KON, BIC, and SEG cells compared to the control group. The effective active concentrations of % cell growth were 5/zM, 15#M, 15# Μ, and 10/zM, respectively (Fig. 1B); however, various concentrations of Phyllanthus emblica were grown on cells of sat, k〇n, BIC, and SEG. There is no obvious inhibitory activity (Fig. lc). There have been 4 studies in the previous study, the combination of urinary hormone and curcumin (hereinafter referred to as compound (1) in the present invention contains 60% catechin and 40% curcumin ), can produce a synergistic anti-cancer effect (Khafif, A. et al. 'as mentioned above; s〇mers-Edgar, TJ et al, as described above). Therefore, it can be seen that the SAT cell line is catechin and turmeric. It is more sensitive than other cell lines, and the SAT cell line is more sensitive to curcumin than catechin (the activity of inhibiting 25% cell growth is 5μΜ and 1〇#Μ, respectively). Because this also the Chinese herbal extract will inhibit the EGFR pathway. (Adachi, s. et al., Cancer Res 67(13): 6493-501, 2007; Chen, A_, J. xu, and Ac Johns〇n,

Oncogene 25(2): 278-87, 2006) ' While in our previous studies, the SAT 201119663 cell line showed significant EGFR in both genes and proteins in other cell lines. It is worth noting that the sensitivity of the SEG cell line flavin is second only to the μ cell line. However, the susceptibility of these four cell lines to Phyllanthus emblica was the same, indicating that E. chinensis itself has no ability to inhibit the growth of cancer cells. Although the effect of Phyllanthus emblica in cell culture is not obvious, the extract of Phyllanthus emblica has (4) oxidative effect, and it has been found to reduce the side effects of chemotherapeutic drugs in clinical application. Phyllanthus emblica L. applied to the dirty brain Human skin fiber _: Fine concentration is finely 5 to Μ (4) ( (This experiment makes φ with 11,5 Pg/mL) (see 卩 Thieves T· et al., as mentioned above). Therefore, we propose a combination of catechin, curcumin and Phyllanthus emblica (hereinafter referred to as compound (2), including 5G% of curcumin, 25% of curcumin, 25% of emblica), and explore its biological effects. In order to test whether the addition of Phyllanthus emblica L. in the present invention contributes to the synergistic inhibition of catechin and curcumin on the growth of cancer cells, catechin 5 〇% inhibiting 25% cell growth and 25% galvanin and 25 μ 余 余 子Add to different cell lines to explore the difference between the compound (2). yulin, curcumin and emblica, and compound (1): catechin and ginger flavin, the growth inhibition ability of oral cancer cells. All experiments were compared to the control group. The method of treating cells and the method of analysis are the same as described above. This experiment was repeated three times. The results from Figure 2 show that 'we can observe on the fourth day after dosing, compound (b) for cell line SAT (Fig. 2), ΚΟΝ (Fig. 2Β), BIC (Fig. 2C) and SEG (Fig. 2D) 'Inhibition of cancer growth by 55 %, 88 %, 63 % and 49 %, respectively. However, the use of the compound (a) in the cell lines SAT, ΚΟΝ, BIC and SEG inhibited the growth of cancer cells by 37%, 88%, 59% and 29%, respectively. In addition, I S1 19 201119663 unilateral use of sulphate, curcumin and amla, inhibited cell growth of 25 / 〇, 25% and 10 % in SAT and SEG, respectively, and inhibited 25 in KON and BIC. 〇, 25% and 5% of the cell lines grew. So we can see

Only, prime! Sustain and Phyllanthus emblica synergistically inhibit the growth of cancer cells. Among them, the results of the SAT spear SEG cell line can be seen that the anti-cancer activity of the compound (b) is better than that of the compound (a) large J 20 /. about. According to the results, Phyllanthus emblica L. can enhance the synergistic effect of catechin and keratin on the growth of cancer cells, so in the clinical treatment, the combination of f-staphylin and curcumin can be reduced. The individual (4) of the two drugs can relatively reduce the side effects and possible drug resistance of the individual. Example. · Containing catechins, curcumin and Phyllanthus emblica L. to promote the apoptosis of cancer cells. The fineness of the fine fertilizer in the cell is extremely dilute, and the normal cells cannot be reduced. Thus, I have a high-egfr_expression of the cell line and other unpublished results), using the formalin-fixed-crystal coloring method to investigate the combination of compound and unilateral inhibition of cell formation in vitro. It can be clearly seen from the figure W that the effect of the combination and the unilateral is poor. Guang 3^ is the quantitative result of staining the cell community with crystal violet. Compared with the use of catechins and turmeric, and yam, it can inhibit Μ%, 〇// and 36% of SAT cells. Interestingly, the use of compound (b) effectively inhibited 91.9% of cell-forming communities. However, the compound (1) can only inhibit the formation of 77.8 % of SAT cells in 201119663. To explore whether the compound (1) and compound (2) inhibit the high EGFR_ expression of SAT cell line cell growth and cell community formation performance, whether through the mechanism of apoptosis, we use AnnexinV staining and cell cycle analysis Further exploration. There are some types of changes that can be used to confirm apoptosis, including, for example, loss of cell membrane symmetry, cytoplasmic and nuclear condensation, and DNA cleavage between nucleosomes. Among them, the loss of symmetry of the cell membrane is an early feature of apoptosis. Phospholipid phosphatidylserine (PS), which is on the cell membrane of early apoptotic cells, turns from the inside of the cell membrane to the surface of the cell membrane. Annexin V is a phospholipid binding protein that relies on calcium ions and has a high binding capacity to serotonin. Therefore, cells with early apoptosis can be marked by FITC (green fluorescent substance) combined with AlexinV. Briefly, 5,000 oral cancer cells were inoculated into a %-well cell culture dish. 'The cell culture medium was replaced with a suitable cell culture medium every other day'. Four days after the drug administration, the Annexin V stain method was used. Sheu, JJ et al., 68(11): ρ·4050-7, 2008), combined with fluorescence microscopy to detect early apoptosis of cancer cell cells. This experiment was repeated three times. The 'sub-Gl phase' represents the result of advanced apoptosis in cells. Since the late stage of cell death caused the DNA to break, it was observed by flow cytometry that the cell cycle phase shifted from the G1 phase to the left to the sub-Gl phase. Inoculate 30,000 to 50,000 oral cancer cells in a 3 cm culture dish. 'Change the cell culture medium to a cell culture medium containing the appropriate ratio of compound cells every other day. Five days after dosing, collect the cells and 21 201119663. The % ethanol was fixed in a -20 〇C refrigerator for one day, and then the cell distribution of the cancer cells sub-Gl, Gl, S, G2/M cell cycle was analyzed by flow cytometry. The results of analyzing the distribution of each cell cycle in ModFit software are shown in Figure 3B and Table 2 below. Figure 3B is a quantification of the results obtained for a total of at least four hundred or more cells from different fields of view. Compared with the control group, it can be seen from Fig. 3B that the compound (1) and the compound (2) were used in the SAT cell line, and an increase of 7.9% and 22.9% of Annexin V positive cells were produced, respectively. The use of catechin, curcumin, emblica and control group alone resulted in 7.2%, 7.1%, 5.8 % and 4.4% AnnexinV positive cells, respectively. Therefore, 'complex (II) promotes the production of more Annexin V-positive cells than the compound (1) due to the synergistic effect. Table 2 below shows the analysis compared to the control group. 2. Analysis of the cell cycle of catechin, curcumin and antioxidants on SAT oral cancer cell lines. Treatment item cell cycle phase % sub-Gl Gl S G2/M Control 7.5 68.8 15.1 8.6 EGCG-lOuM 8.1 72.3 13.2 6.4 Curcumin-5 uM 9.2 71.5 12.7 6.6 Amla-25 uM 8.8 69.2 13.3 8.7 Recipe 1 6,9 64.7 19.1 9.2 Recipe 2 9.0 51.6 27.6 11.7 The results showed that the compound (b) caused a slight increase in the SAT cells by 1.5% in the sub-Gl phase, which significantly reduced the gi phase by 17.2% with a significant increase in the S/G2M phase of 15.6%. 22 201119663 One's compound (a) slightly increased SAT cells staying in the S/G2M phase 46〇/. . The unilateral use of sputum, sulphate and Phyllanthus increased the sub_G1 phase 〇.6/〇, 1.7% and 1.3% ’, and G1 phase 3 5%, 2 7% and 〇 4%, respectively. Therefore, the rear (2) SAT cells stayed in the s/G2M phase due to the Lang effect, but there was no obvious sub-Gl phase rise. Many literatures have pointed out that the anticancer mechanism of catechin and curcumin is accomplished through the pathway of apoptosis. Therefore, we evaluated the early and late apoptosis by eight sputum V staining and cell cycle analysis experiments. The results showed that the compound (1) compared with the compound () induced more SAT cell lines to produce Annexin V staining positive cells, and caused the cell cycle to stay in the S/G2M phase and the sub-G1 phase. Among them, compared with the pair, the compound (2) increased 18.5% of the eight (10) sputum v staining positive cells, increasing 15.6% of S/G21V [phase cells and 1.5% of sub-Gl phase cells. The results showed that the use of unilateral results caused the staining results of AnnexinV to be less than that of the control group; however, compared with the control group, the unilateral use of catechin, curcumin and Phyllanthus emblica mainly caused the cancer cells to stay in the G1 phase and the sub-G1 phase. Previous studies have shown that 5.5 catechins and 5.0 curcumin act on the pre-cancerous cell line for 24 hours, and the effect of the use of tyrosin alone is in the G1 phase, while the effect of curcumin is in the S/G2M phase. However, in the breast cancer cell experiment, 2〇#Μ catechin combined with 3.0/ζΜ of the oxytocin for 12 hours, the breast cancer cells stayed in G2/]y [phase. However, the experimental results of the embodiments of the present invention are similar to the previous research results, and more illustratively, the additive effect of the Chinese medicine composition of the present invention inhibits the cancer effect, which can be accompanied by changes in the cell cycle phase, and promotes cancer cell death. Death happened. [S] 23 201119663 Example (5): Containing silk fibroin, 4 flavin and Phyllanthus _# composition inhibition Tumor growth of cancer xenograft nude mice

It can be seen from the experiments of the in vitro cell lines of the first and second embodiments that the compound (II) has synergistic effect on inhibiting the growth of cancer cells and promoting apoptosis compared with the compound (1). Therefore, in the present example, the effect of the composition of the present invention on inhibiting the formation of cancer cells in vivo was further carried out in a compound (2) xenograft nude mouse of SAT and SEG. In order to investigate the ability of the compound to inhibit tumor growth in living organisms, we used BALB/c nu/nu male nude mice as SAT and SEG for oral cancer cell xenografts, first with catechin 25〇mg/j^/ Days (SEG-inoculated SEG mice were changed to a dose of 500 mg/kg/day); curcumin 2 mg/kg/day; Phyllanthus 250 mg/kg/day; compound (b) catechins 5%, Curcumin 25% and Phyllanthus emblica 25%; 0.1% DMSO 10ml/kg/day was used as a control group for 5 weeks to activate the immune system of nude mice. Then, 4 million SATs and 1 million SEGs were inoculated in a 1:1 manner, and the matrigel was mixed and inoculated into the skin of nude mice. The recipe was continued every day until the SAT tumor size was approximately 1 mm 3 (approximately 4 days); the seg tumor was sacrificed at approximately 1500 mm 3 (approximately 25 days) and the tumor was sacrificed. Tumors were fixed by ίο % formalin and stained with H&amp;E staining and Ki67 immunohistochemistry to determine whether natural plant extracts inhibit tumor cell proliferation. During the period before the sacrifice, the tumor size was recorded with a measuring rule every two days, and the nude mice were observed for abnormalities. When the animal is sacrificed, the liver and kidney are fixed with 10% formalin and stained with H&amp;E to determine whether liver or kidney damage will occur during the administration of the drug. 24 201119663 From Figure 4 (A, B) compared with the control group As a result, it was observed that the tumor growth curve and tumor size were significantly decreased in the SAT group nude mice at the 3rd week after the tumor injection; conversely, the SEG group was only slightly reduced (results not shown). Since the proliferation rate of cancer cells is quite fast, we evaluated the efficacy of compound (2) in inhibiting the proliferation of cancer cells by the amount of Ki_67. In Fig. 4(c), the results of immunostaining staining can be seen in comparison with the control group, indicating that the yarn (2) can inhibit cancer cell proliferation. Therefore, similar to the experimental results of the aforementioned cell strains, the compound (II) successfully inhibited tumor growth and cell proliferation. Example 5: The molecular sieve of the composition containing catechin, curcumin and Phyllanthus emblica L. was studied by cDNA microarray analysis. The results of the animal experiment of the above Example 4 were found, and the compound (2) was used for the SAT. Xenografted nude mice have efficacy, and in cell line experiments, KON has been shown to be effective against compound (b). Therefore, in order to explore what mechanism is included in the compound

In λ oral therapy, cDNA microarray (AFFYMETR1X U133-2 biochip) was used for analysis, and compound (b) was turned on or off (tum 〇ff) for certain specific genes, and From the perspective of gene expression, we explored the molecular mechanism of the tumor suppressor molecule in the compound (2), in order to sort out the molecular pathways involved in the compound (2) treatment. Ten million SAT and KON oral cancer cells were inoculated into 15em culture 011, and the cell culture medium was replaced with a cell culture medium containing catechin ΙΟμΜ, curcumin 5μΜ and 25 201119663 余甘子 25μΜ compound (2). The SAT cell line was treated and cells were harvested 48 hours after dosing and stored in the RNA/α stage. It is first placed in liquid nitrogen and then transferred to -80 ° C for storage. The mRNA is then extracted and inverted into cDNA, followed by subsequent cDNA microarray analysis (Jimeno, A. et al., supra). This experiment was repeated twice. The results are shown in Table 3 below. Table 3. Whole-genome profiling analysis of SAT cells after compound (B) treatment Gene name CCL4 EPHB2 CASZ1 ADAMTS7 SDCCAG8 ZNF354C SLC2A5 OR7A10 ATP8B2 COX6A1 Accession number AK024994 AK025173 AK021990 NM.014272 AK024613 AL137483 NM_003039 AC005255 AL137537 AF020589

Ff〇W changes a 119:08 -12.82 -3.28 -3.07 •2.4 -2.29 -2.17 -2.17 -2.11 -2.04 Description CCL4: Chemokine (C-C motif) Ligand 4 EPHB2: EPH Receptor B2

CASZ1: Castor homolog 1, zinc finger (fruit rope) ADAMTS7: ADAM metalpeptidase has type 1 thrombospondin motif, 7 SDCCAG8: serologically identified colorectal cancer antigen 8 ZNF354C: zinc finger protein 354C SLC2A5. /Cereal carrier steroid 2 (promoting glucose/fructose transporter), member 5 'OR7A10: olfactory receptor, family 7, subfamily a, member 1〇ATP8B2: ATPase, class I, class 8B, member 2 C0X6A1: cytochrome c oxidase subunit VIa polypeptide j ///cytochrome c oxidase subunit VIa polymorphic i FGFR2

PBX2 MGC34824 RBPMS HNRPM SNAI1 LGALS1 KCNK15 DERL3 VM NM.022976 BC003111 AK023697 NM_006867 AK022050 NM-005985 M14087 NM_022358 AI655697 AI922599 -2 -2 -1.99 -1.98 -1.95 -1.94 -1.87 -1.83 -1.82 -1.81 The team is noisy Factor receptor 2 ϊCell growth factor receptor, Gu bone craniofacial dysplasia sin syndrome, Jason _ Weiss incomplete 1' PBX2: 刖-B-cell leukemia transcription factor 2 MGC34824: hypothetical protein MGC34824

KbPMb: KNA-binding protein, riboprotein M with multiple HNRPM·heterologous nucleus SNAI1: snail homolog 1 (fruit jar) LGALS1: lectin, galactoside-binding KCNK15··potassium channel, subfamily Ruler, ' DERL3: Derl-like functional domain family into 'VIM: vimentin, soluble, Ugalectin 1;) 26 201119663

DEPDCl NM—017779 -1.79 DEPDCl: DEP domain contains 1 CDC2L6 AY028424 -1.78 CDC2L6: cell division cycle 2 - 0 (CDK8 - class) / / / cell sorting cycle 2 - 6 (CDK8 - class) &lt; PLCH1 AK022610 -1.77 PLCH1: Phospholipase C, etal HAS3 AI739514 -1.77 HAS3: Hyaluronan Synthase 3 LAMP3 NM_014398 -1.76 LAMP3: Lysosomal-Linked Membrane Protein 3 MMP10 NM.002425 -1.76 MMP10: Matrix Metal Peptidase ι 〇 (Matrix lysin 2) GM2A M76477 -1.76 GM2A: GM2 ganglion glycoside activator TncRNA AV699657 -1.76 TncRNA: trophoblast-derived non-coding coffee A C0L1A2 AA788711 -1.75 (:0〇8: collagen, first Type 1, &amp; 1 coffee 2 GRPR NM_005314 -1.75 GRPR: gastrin-release peptide receptor MX1: mucovirus (influenza virus) resistance 1, interferon-inducible egg MX1 NM.002462 -1.69 white p78 (mouse) /// Mucin virus (influenza virus) resistance 1 interferes with the serotonin protein p78 (mouse) ! ^ CLEC3B NM_003278 -1.69 CLEC3B: C-type lectin domain family 3, member b IGFBP7 NM-001553 -1.67 IGFBP7: Insulin-like growth factor binding protein 7 TAS2R7 NM.023919 -1.65 TAS2R7: taste receptor Type 2, member 7 hCAP-H2 U62317 -1.65 hCAP-H2: kleisin beta PCF11 AW118175 -1.63 PCF11.PCF11, cleavage and polyadenylation factor subunit homolog (s cerevisiae), v ' APOH AV648193 -1.63 APOH : apolipoprotein H (beta-2·glycoprotein I) TRIM22 AA083478 -1.62 TRIM22: triplet motif-containing 22 ZNF447 AI375002 -1.61 ZNF447: zinc finger protein 447 MY03A NM-017433 -1.61 MY03A: myosin progenitor AOAH AK026853 -1.61 AOAH: Stable Oxygen-based Hydrolase (neutrophil) PSMB9 NM_002800 -1.6 PSMBf: Proteasome (prosome, m _ 9 (large multifunctional enzyme 2) flat ' _ S, CDGAP AB033030 -1.59 CDGAP : Cdc42 GTPase-activating protein LOC642776 BC003645 -1.59 LOC642776: hypothetical protein LOC642776 DTNB AI672185 -1.57 DTNB: dystrobrevin, beta CHD9 AW300405 1.6 CHD9: color functional domain helicase DNA binding protein SYNCRIP AI472757 1.61 gCRIP: fine binding protein binding , cell interactions IQGAP1 D29640 1.63 IQGAP1: Yangca activated protein with IQ motif} FXR1 AI990766 1.64 FXR1: Fragile X smart retardation, often stained SCAMPI BF058944 1.64 SCAMPI : Secretory carrier prion protein 1 SMC3 AI373676 1.66 SMC3: Structure of chromosome 3 maintenance USP10 BC000263 1.69 USP10: ubiquitin-specific peptidase 10 TMEM106B AV705186 1.7 TMEMKteB··transmembrane protein 1〇6B BICD2 BC002327 1.71 BICD2: bicaudal D homolog 2 (fruit) CDC42BPA NM—003607 1.72 CDC42BPA: CDC42 binding protein kinase _ _κ-like) WASL BE504979 1.73 WASL: Wiskott-Aldrich syndrome group. Similar 27 201119663 BRCC3 S72931 PRO1073 AA827878 ZFP91 AA758013 OVOLl AA588400 NOLCl AI355279 GNG12 N32508 FAM60A BG479856 RH0BTB3 N21138 SPIN iNM_0067l7 NFAT5 NM-006599 CALDl AI685060 CPSF2 AA583986 VIL2 AF199015 ACBD3 AI636775 ILF3 BG032366 GAPVDl AI797397 MALATl W80468 TOPI AW025108 FOXLl AF315075 ZWILCH NM_0l7975 USPl AW499935 MY06 AA877789 SEC23IP AK021846 NBN AI796269 NIPSNAPl AW083371 DNAJC3 NM_006260 UBXD8 BF116183 UHMKl AW173222 CX3CR1 U20350 ILIRN AW083357 ZC3H7A AW169959 CEP57 AI123527 PTPLB AI813654 SON AA664291 PRO1073 NM—014086 RNF19 NM—015435 NBN A K001017 74 4 4 4 5 5 5 7 7 1 2 2 4 5 5 7 8 1 3 3 4 5 5 8 9 7- 7W 7W 7W 7W 7W 7W 8. 8· οςοςοςος oq ny 9* 9- 9* 9· lx :i lx 1i lx -IX *'—ΊΑ Ί · 1X - · · · 1A ΊΧ -1X · -^ lx .—«423 5 6 Π ~ un J ui 4 Λ ▲ ΊΧ Tx oi oi l oi 7- -- · VO oo 2· 2 2 2 2 l BRCC3: complex containing BRCA1/BRCA2-, subunit 3 PRO 1073: PRO 1073 protein ZFP91: zinc finger protein 91 homolog (mouse) OVOLl:ovo-similar 1 (fruit! 13⁄4) N0LC1: nucleolar and spiral wound body phosphoprotein 1 GNG12: guanosine nucleoside binding protein (G protein), gammal2 FAM60A: family with sequence subfamily 6〇, member a RH0BTB3: containing Rho-related BTB domain 3 SPIN: spindle protein NFAT5: nuclear factor 5 of activated T-cells, tension-reactive CALD1: calmodulin-binding protein 1 CPSF2: cleavage and polyadenylation specificity, f-factor 2 l〇〇kDa VIL2: woven hair protein 2 (ezrin) ACBD3: containing the brewing-coenzyme a binding domain 3 ILF3: interleukin-enhancing protein binding factor 3,9〇kDa GAPVD1: GTPase activating protein and vps9 functional tumor Metastasis related to lung adenocarcinoma丨 (non-coding ΤΟΡ1: topoisomerase (DNA) I F0XL1: fork tip frame region L1 ZWILCH: Zwilch, kinetochore associated, homolog (fruit rope) USP1: ubiquitin-specific peptidase 1 MY06: muscle Globulin VI SEC23IP: SEC23 interaction protein NBN: rnbrin: nipSnap homolog i (c. degans) DNAJC3. DnaJ (Hsp4〇) homolog, subfamily gg g 3 UBXD8: UBX domain containing 8 5 UHMK1 : U2AF homologue motif (u^) stimulating CX3CR1: chemokine (C_X3_C motif) IL1RN: interleukin-1 receptor antagonist bottle 1 ZC3H7A: containing zinc finger CCCH-type 7A CEP57: centrosome Protein 57kDa egg if (four) clock two enzymes (hybrid generation catalytic SON: SON DNA binding protein PR〇1073: PR〇l〇73 protein RNF19: ring finger protein 19 NBN: nibrin 28 201119663

RCP9 NM—014478 2.29 GAPVD1 AK021533 2.3 CBX5 L07515 2.31 MTUS1 BE552421 2.33 DNMT3A NM—022552 2.34 RRBP1 NM—004587 2.35 ΙΠ16 BG256677 2.38 LIPH AA088857 2.38 M0BK1B NM—018221 2.4 ACTR2 AA699583 2.44 DSC2 BF196457 2.49 TRIB2 BC002637 2.5 PRO1073 AF113016 2.51 EPRS AI142677 2.58 ID2 NM_002166 2.58 MAP4 AI553791 2.6 RAPGEF5 AI263837 2.6 KIAA1333 AA887053 2.6 TA0K1 AB037782 2.61 USP10 BG390445 2.63 PAP0LA AI670847 2.64 ASPM AK001380 2.68 EHBP1 BF116032 2.74 BCLAF1 BE963370 2.76 MAP4K5 Z25426 2.79 PSME4 AI972268 2.82 VIL2 BF663141 2.85 CCDC98 AK021582 2.89 BAT2D1 AI359472 2.9 CPNE3 AA541758 2.92 THRAP3 BE967048 2.97 EIF4G1 BE966878 3,06 RBM8A AF182415 3.15 PPFIBP1 AI962377 3.28 ZNF294 AK023499 3.33 TPM4 BC002827 3.35 IQGAP1 AI679073 3.49 Activator subunit 4 RCP9: vasopressin-related receptor composition protein GAPVm: GTPase activating protein and cleavage domain! CBX5: chromosomal framework homolog MTUS1: mitochondrial tumor suppressor protein 1 methyltransferase 3 _ a protein 1 homolog 180kDa (dog) let 丄 1 &amp;: sesame, gamma-inducible white] 6 LIPH: lipase, member η = 3: MQB1, class - MPS - binder mer activator 1 Β = 13⁄4 white-related protein 2 homolog (yeast) TRIB2: tribbles homolog 2 (fruit butterfly) PR01073 :PR〇1〇73 Protein EPRS: Glutamine-Based Amidyl-A Synthetase Inhibitor of DNA Binding 2, Dominant Negative Helix, Ring Protein MAP4: Microtubule-associated Protein 4 RAPGEF5. Rap Guanine nuclear bitter g吏 exchange factor (gef) 5 KIAA1333: KIAA1333 ΤΑΟΚ1: 激酶 Kinase 1 USP10: ubiquitin-specific peptidase 1〇PAPOLA: poly(A) synthetase aipha ASPM: asp (abnormal spindle)-similar, Micro-related (fruit rope) EHBP1: EH domain binding protein j BCLAF1: BCL2-related transcription factor i MAP4K5: mitogen-activated protein kinase 5 PSME4. protein spider (pr〇s〇me, maeiOpaiii&gt; VIL2: Chorionic protein 2 (ezrin)

CCDC98: Contains the coil-helix domain 98 BAT2D1: Contains the BAT2 domain 1 CPNE3: copine III THRAP3: scorpion hormone receptor-associated protein 3 EIF4G1: eukaryotic translation initiation factor 4 gamma ! RBM8A: RNA-binding motif Protein 8A 'PPFIBP1: PTPRF interaction protein, binding protein 1 (liprin beta 1) ZNF294: zinc finger protein 294 TPM4: tropomyosin 4 IQGAP1: QTPase-activating protein 1 containing IQ motif [S] 29 201119663 LIMA1 BF197289 3.49 LIMA1: LIM domain and actin binding j ZFR BF445199 3.53 ZFR: zinc finger RNA binding protein STON2 BE379761 3.54 STON2: Stonin 2 MBNL2 NM_005757 3.64 MBNL2: class-muscleblind 2 (fruit! m) MALAT1 AF132202 3.67 MALAT1: tumor metastasis Related lung adenocarcinoma transcripts 非 (non-RNA) OSBPL8 AW978375 3.74 0SBPL8: oxy-sterol-binding protein 8 CDC27 AI203880 3.77 CDC27: cell division cycle 27 EEA1 AI916242 4.13 EEA1: early endosomal antigen i, i62kD MALAT1 AW005982 4.24 MALAT1: Μ 移 之 肺 赫 赫 非 (non-RNA) with 迮HRB AI742626 4.56 HRB: HTV-1 Rev binding protein DHX9 BE561014 4.87 DHX9: DEAH (As p-Glu-Ala-His) box polypeptide 9 FAM44A AI049791 5.51 FAM44A: family with sequence similarity 44, member a ATP13A3 NM a 024524 5.93 ΑΤΠ3Α3: ATPase type 13A3 ZFR AI459274 6.53 ZFR: zinc finger RNA binding protein _ SUPT16H AK024072 6.62 SUPT16H: The sequence of the Ty 16 homologue n See Table 2 for the results. Of the 168 genes with significant expression variation, 146 genes are known genes. Many known oncogenes, such as EPHB2, FGFR2, PBX2, etc., are inhibited by compound (b) treatment. Conversely, some genes that inhibit the growth of cancer cells, such as BCLApi, MAP4, MTUS1, etc., are shown in large numbers. If analyzed by the signal transmission network, it can be found that the signal transmission pathway in which the most genes participate is the enhancement of the cell attachment ability #, which inhibits the spread of cancer cells and promotes the differentiation of cancer cells, and causes the cancer suppressive regulation of contact inhibition between cancer cells. Based on the above results, the present invention combines Phyllanthus emblica with catechin and curcumin (complex 2), and indeed has the cancer suppressing effect of the combination of catechin and curcumin (combination 1) and results in animal experiments. It is clearly demonstrated that Compound (b) is effective in the prevention and treatment of cancer 'as expected to inhibit tumor size and tumor proliferation 30 201119663 (Fig. 4A-C). Previous studies have shown that catechin and curcumin can inhibit the growth of breast cancer by inhibiting the expression of the vascular receptor-1 protein (Somers-Edgar, T.J. et al., supra). The angiogenic factor receptor _丄 (VEGFR-1) or Flt-Ι is involved in the development of cancer formation, inflammatory response, angiogenesis, and tumor migration (Ponticdli, S. et al., jBiol Chem, 2〇〇8). In order to identify a specific gene or pathway that is related to the anti-cancer efficacy of the compound (b), I am equal to the analysis of the gene in the fourth embodiment of the compound (2) for the SAT cell line. As a result, it was found (Table 2) that the action of the compound (II) of the present invention can activate the signal pathway of the signal molecule of nectin, and the function of the nectin message in recent studies is considered to regulate the cell attachment ability and inhibit the spread of cancer cells. Promotes the differentiation of cancer cells and causes a tumor suppressive effect of contact inhibition between cancer cells (Takai, γ. et al. Nat Rev Mol Cell Biol 9(8): 603-15, 2008), and thus the present invention contains The traditional Chinese medicine composition of quercetin and Phyllanthus emblica extract has the effects of inhibiting the spread of cancer cells and promoting the differentiation of cancer cells, and can be applied to treat or prevent tumor metastasis. Other Aspects All of the features disclosed in the 5th book can be combined in any combination. Each of the functions disclosed in this specification can be replaced by alternative features of the same, equal or _ purpose. Accordingly, the features disclosed are merely examples of a series of equivalents or similar features, unless otherwise indicated. From the foregoing description, the basic features of the present invention can be easily determined by those skilled in the art, and various changes and modifications of the present invention can be made to make it suitable for various differences. Use and condition. Therefore, other specific aspects are included in the scope of application for patents. All references cited in this article, including publications, patent applications, and patents, are fully referenced in this article, and are obtained separately from each reference or _ are designated as To the same extent, and as described in its entirety (to the maximum degree permitted by law). All standards

The subject matter and the sub-standards are for convenience and are not limited to the present invention. [Simple description of the figure] Figure 1 shows the active concentration of Chinese herbal extracts to inhibit cancer cell growth. Among them, fig, B and C represent different concentrations of catechin, curcumin, and emblica produce inhibitory cells against 5,000 different oral cancer filaments (mouth is SAT; _k〇n; _ for ship; _ BIC) The ability to grow. Four days after drug treatment, the sybrgreen method found that these three drugs can reduce the growth of 25% cancer cells.

Figure 2 shows that quercetin and amlaline can synergistically inhibit the growth of cancer cells. Two of them 2A, B, C and D represent the concentration of catechins and quercetin, which inhibit the growth of 25% cells, respectively. Yuganzi of _ explored its ability to inhibit growth of 5,000 (4), KON, BIC and SEG oral cancer cells. Four days after drug treatment, the ability of different compounds to inhibit cancer cell growth was found by sybrgreen paste. Figure 3, the member is not a nutrient, the amount of vapor and Phyllanthus emblica can synergistically inhibit the formation of cancer cells and promote cancer cell cells; Figure H ·· 20,000 pieces of torn oral cancer 32 201119663 ,, the seed is inoculated in the T25 flask, the next day the cell culture medium is replaced with a suitable proportion of the compound cell culture solution, the seventh day after the drug is replaced - The cells containing the same drug were cultured overnight, and finally the cell culture solution was blotted dry on the fourteenth day after the administration of the drug: I liter 10/.曱 ’ 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛 洛After the flask is naturally air-dried, take a photo with the camera; Figure 3Α-2: Calculate the change in the number of cancer cells in the culture bottle to which different drugs are added. Figure 3Β: Inoculate 5,000 oral cancer cells in a % well cell culture dish, and replace the cell culture medium with a cell culture solution containing the appropriate ratio of compound cells every other day. After four days of administration, the Annexin V staining method is combined with a fluorescence microscope. Use 'to detect early apoptosis in cancer cells. Figure 4 shows that Compound II inhibits tumor growth in cancerous xenograft nude mice. The catechins were 250 mg/kg/day (the nude mice inoculated with SEG were changed to 500 mg/kg/day); curcumin 2〇〇mg/kg/day; Phyllanthus 250mg/kg/ Day; catechin 50% + curcumin 25% + emblica 25%; 0.1% DMSO for the right, group, and the first preventive action for 5 weeks to activate the nude mouse immune system, and then inoculate 4 million SAT One million SEG mixed matrigels were inoculated subcutaneously in nude mice, and then continued to feed the formula daily until the SAT tumor size was about 15 mm3 (about 35 days); when the SEG tumor size was about 15 mm3 (about 14 days) The nude mice were sacrificed and the tumors were removed for tissue molecular identification. (A) is a tumor growth curve; (B) is a tumor size difference map; (C) is a Ki-67 immunohistochemical staining result. I S1 33 201119663 [Main component symbol description]

Claims (1)

201119663 VII. Patent Application Scope A proprietary Chinese medicine for the treatment of cancer caused by cancer, which contains 6% of catechins, 6G% of the extracts of the yellow cups, and miscellaneous Accepted _, rare or excipients, the proportion of each component is adjusted according to different cancer types. 2. 3. According to: The composition of the Chinese medicine as described in item i of the patent scope, wherein the cancer is a cancer having an EGFR expression. According to the patent application No. 1 smear composition, the cancer is oral cancer. 4. The root shot please use the special fiber to cover the towel of the first one, and the individual is human. 5· - A pharmaceutical composition that promotes the growth of cancer cells, which contains lignin 2〇/0 to 6〇/ in children up to 6〇%. The amount of flavin and 2% to % of the yam extract> and pharmaceutically acceptable _, diluent or excipient, wherein the proportions of each component are adjusted according to different cancer types. 6. The medical composition according to the patent application scope 5th tone μ, +, #, wherein the cancer cell is a cancer cell having high EGFR expression. 7. The pharmaceutical composition according to claim 5, wherein the cancer cell is an oral cancer cell. ★ The species is used to suppress the miscellaneous n, which contains (10) to the secret lignin, 2〇/0 to 6〇% of flavonoids and Shi Zhisheng. The extract of Ganzi, 1 western medicine can be charged with carrier, diluent or excipient, and the proportion of each component is adjusted according to different cancer types. 201119663 9. The pharmaceutical composition according to item 8 of the scope of the patent application, which inhibits the spread of cancer cells and promotes the differentiation of cancer cells by regulating cell attachment ability, thereby causing contact inhibition between cancer cells. 10. The pharmaceutical composition according to claim 1, 5 or 7, wherein the pharmaceutical composition comprises 20% to 60% catechin, 20% to 40% curcumin and 20% to 40% The remaining nectar extract. 11. The pharmaceutical composition according to claim 1, 5 or 7, wherein the pharmaceutical composition comprises 50% catechin, 25% curcumin and 25% geranium extract. 36