A cancer treating composition derived from psoralea corylifolia and epimedium

Abstract

The present invention relates to a composition for treating cancer, the composition comprising a component derived from psoralea corylifolia and a component derived from epimedium, such as psoralen A and epimedin II, epimedin B or epimedin C. The present invention conducts component mining on psoralea corylifolia and epimedium, and determines the effect of the components in psoralea corylifolia and epimedium on synergistic treatment of cancer. The composition of the present invention inhibits cell proliferation for various common cancers such as liver cancer, lung cancer, breast cancer, etc., so it can be effectively used for treating cancer.

Description

Technical Field

[0001] The present application belongs to the field of natural medicine and tumor treatment. Specifically, the present application provides a composition for treating cancer derived from Psoralea corylifolia and Epimedium. Background Art

[0002] Epimedium and Psoralea corylifolia are commonly used tonic Chinese medicines. Both are pungent and warm in nature, and can warm and tonify kidney yang. They are often used in combination with other tonic medicines in prescriptions for the treatment of kidney yang deficiency, liver and kidney deficiency, lung and kidney qi deficiency, or spleen and kidney yang deficiency. Psoralea corylifolia has the effects of warming the kidney and supporting yang, warming the spleen and stopping diarrhea, and is a commonly used Chinese medicine for the treatment of kidney and spleen yang deficiency. Psoralen is a coumarin-like and coumarin-related benzofuranoside with antioxidant activity, antitumor effect, and antibacterial activity. Studies have found that psoralen can inhibit the proliferation of human liver cancer SMMC-7721 cells, human breast cancer MCF-7, and MDA-MB-231 cells, and promote cell apoptosis, suggesting that psoralen may be a new treatment for the prevention and treatment of liver cancer. In summary, Psoralea corylifolia has great potential for treating tumors, and further research is necessary to explore the anti-tumor effect of Psoralea corylifolia.

[0003] Epimedium is an important medicine for warming yang and tonifying kidney. Studies have found that epimedium can inhibit the migration of mononuclear malignant glioma cells. Icariin is a compound extracted from plants of the genus Epimedium. Studies have found that ICA induces differentiation of B16 mouse melanoma cells by inhibiting the ERK1 / 2-p38-JNK-dependent pathway, causing cell cycle arrest in the G0 / G1 phase. Studies have found that icariin can induce apoptosis of xenograft ovarian cancer cells and inhibit tumor growth. Studies have found that icariin exhibits anti-tumor effects both in vivo and in vitro in P815 tumor-bearing mice, and can upregulate LMP / TAP-related molecules and induce MHC-I expression, thereby enhancing immune surveillance and alleviating tumors, indicating that icariin may become an effective antigen adjuvant for tumor treatment. The above studies show that epimedium is expected to provide clues for related tumor treatment and provide new opportunities for future tumor treatment. Summary of the invention

[0004] Since the chemical substances in Psoralea corylifolia and Epimedium brevicornum that induce cancer cells are not clear, it is necessary to screen the anti-tumor active ingredients of Psoralea corylifolia and Epimedium brevicornum. This study explored the ingredients of Epimedium brevicornum and Psoralea corylifolia to clarify the material basis of the synergistic treatment of cancer by the ingredients in Psoralea corylifolia and Epimedium brevicornum, in order to provide guidance and support for the prevention and treatment of cancer with Psoralea corylifolia and Epimedium brevicornum, and lay a foundation for guiding the development of original new Chinese medicines and rational clinical use of drugs.

[0005] In one aspect, the present application provides a composition for treating cancer, comprising a component derived from Psoralea corylifolia and a component derived from Epimedium brevicornum.

[0006] Furthermore, the ingredient derived from Psoralea corylifolia is selected from one or more of bakuchiol, psoralen, psoralen glycoside, isopsoralen, psoralen, isopsoralen, isopsoralen dihydroflavone, neopsoralen isoflavone, psoralen dihydroflavone methyl ether, psoralen and psoralen B.

[0007] Furthermore, the ingredient derived from epimedium is selected from one or more of icariin, icariin, icariin I, icariin II, epimedin A, epimedin A1, epimedin B, epimedin C, dehydrated icariin and icariin A.

[0008] Furthermore, the component derived from Psoralea corylifolia is psoralen A, and the component derived from Epimedium is icariin II, epimedin B or epimedin C.

[0009] Furthermore, the active ingredients in the composition are psoralen A, icariin II, epimedin B or epimedin C; preferably, the composition does not contain other active ingredients.

[0010] Furthermore, the weight ratio of psoralen to icariin II, epimedin B or epimedin C in the composition is 8:1-1:2.

[0011] Furthermore, the weight ratio of psoralen to icariin II, epimedin B or epimedin C in the composition is 4:1-1:2.

[0012] Furthermore, the weight ratio of psoralen A to icariin II in the composition is 2:1; or, the weight ratio of psoralen A to epimedin B in the composition is 2:1; or, the weight ratio of psoralen A to epimedin C in the composition is 1:1.

[0013] On the other hand, the present application provides the use of the above composition in the preparation of a drug for treating cancer.

[0014] Furthermore, the cancer is liver cancer, lung cancer, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, kidney cancer, thyroid cancer, pancreatic cancer, prostate cancer and bladder cancer.

[0015] Furthermore, the cancer is liver cancer.

[0016] The composition of the present invention is in any clinically acceptable dosage form, including various dosage forms for oral and parenteral administration. For oral administration, it can be tablets, capsules, soft capsules, oral liquids, syrups, granules, dripping pills, orodisintegrating tablets, sustained-release tablets, sustained-release capsules, controlled-release tablets, controlled-release capsules, etc.; for parenteral administration, it can be water injection, lyophilized powder injection, sterile powder injection, infusion, etc.

[0017] Those skilled in the art can use various pharmaceutically acceptable carriers or excipients to construct the above-mentioned dosage forms, such as solvents, co-solvents, pH adjusters, osmotic pressure regulators, stabilizers, antioxidants, fillers, binders, lubricants, disintegrants, coating agents, sustained-release agents, controlled-release agents, adsorption carriers, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1A The cell inhibition rate of different concentrations of psoralen is shown;

[0019] Figure 1B The cell inhibition rate of different concentrations of bakuchiol is shown;

[0020] Figure 1C The cell inhibition rate of different concentrations of psoralen is shown;

[0021] Figure 1D The cell inhibition rate of different concentrations of isopsoralen is shown;

[0022] Figure 1E The cell inhibition rate of different concentrations of psoralen is shown;

[0023] Figure 1F The cell inhibition rate of different concentrations of isopsoralen is shown;

[0024] Figure 1G The cell inhibition rate of different concentrations of psoralen is shown;

[0025] Figure 1H The cell inhibition rate of different concentrations of psoralen is shown;

[0026] Fig. 1I The cell inhibition rate of different concentrations of isopsoralen dihydroflavone is shown;

[0027] Figure 1J The cell inhibition rate of different concentrations of new psoralea isoflavones is shown;

[0028] Figure 1K The cell inhibition rate of different concentrations of psoralea corylifolia is shown;

[0029] Figure 2A The cell inhibition rate of icariin and icariin is shown;

[0030] Figure 2B The cell inhibition rates of icariin I and icariin II are shown;

[0031] Figure 2C The cell inhibition rates of epimedin A and epimedin A1 are shown;

[0032] Figure 2D The cell inhibition rates of epimedin B and epimedin C are shown;

[0033] Figure 2E The cell inhibition rates of dehydroicariin and icariin A are shown;

[0034] Figure 3A The cell inhibition rate of the combination of psoralen and 8.33 μM Epimedium monomer is shown;

[0035] Figure 3B The King's formula q value of the combination of psoralen and Epimedium monomer at 8.33 μM is shown;

[0036] Figure 3C The cell inhibition rate of the combination of psoralen and 16.67 μM Epimedium monomer is shown;

[0037] Figure 3D The King's formula q value of the combination of psoralen and Epimedium monomer at 16.67 μM is shown;

[0038] Figure 3E The cell inhibition rate of the combination of psoralen and 33.33 μM Epimedium monomer is shown;

[0039] Figure 3F The King's formula q value of the combination of psoralen and Epimedium monomer at 33.33 μM is shown;

[0040] Figure 3G The cell inhibition rate of the combination of psoralen and 66.67 μM Epimedium monomer is shown;

[0041] Figure 3H The King's formula q value of the combination of psoralen and Epimedium monomer of 66.67 μM is shown;

[0042] Figure 4A It shows the cell inhibition rate of psoralen A combined with epimedin B;

[0043] Figure 4B It shows the cell inhibition rate of psoralen A combined with icariin II;

[0044] Figure 4C It shows the cell inhibition rate of psoralen A combined with epimedin C;

[0045] Figure 4D The King's formula q value of psoralen A combined with epimedin B, epimedin II, and epimedin C is shown;

[0046] Figure 5A To show the effect of psoralen A combined with icariin II on tumor mass;

[0047] Figure 5B To show the effect of psoralen A combined with icariin II on tumor volume;

[0048] Figure 5C Shows the effect of psoralen A combined with icariin II on the body weight of mice;

[0049] Figure 5D Shows the effect of psoralen A combined with icariin II on organ coefficients. DETAILED DESCRIPTION

[0050] Example 1 Materials and Basic Methods

[0051] 1. Cell Culture

[0052] HepG2 cells react similarly to primary mouse hepatocytes and primary human hepatocytes. The culture medium used for HepG2 cells is DMEM medium containing 10% fetal bovine serum and 1% penicillin-streptomycin, and cultured in a humidified incubator at 37°C with 5% carbon dioxide. This experiment used cells from passages 3 to 7 for the experiment, and only cells with a survival rate greater than 90% were used.

[0053] 2. Drug Preparation

[0054] Eleven monomer components of Psoralea corylifolia were selected for screening, namely, bakuchiol, psoralen, psoralen, isopsoralen, psoralen, isopsoralen, isopsoralen, neopsoralen isoflavone, psoralen methyl ether, psoralen and psoralen B, and 10 monomer components of Epimedium, namely, icariin, icariin I, icariin II, epimedin A, epimedin A1, epimedin B, epimedin C, dehydrated epimedin and epimedin A. Accurately weigh an appropriate amount of each reference substance, dissolve it with cell-grade DMSO, and prepare a mother solution with a concentration of 50 mg / mL, and store it at -80°C for use.

[0055] 3. Cytotoxicity Assay

[0056] HepG2 cells were collected at 9×10 4 The cells were inoculated in a 96-well plate at a density of 100 cells / well, and the cells were cultured in a complete medium in an incubator for 24 hours before administration. After the cells were exposed to the drug or DMEM for 24 hours, the absorbance (A) was detected at 450nm using a CCK8 kit and a microplate reader. The experiment was repeated 3 times. The inhibition rate of the drug on HepG2 cells was then calculated according to the formula. The inhibition rate of the cells was calculated according to the following formula.

[0057] Cell inhibition rate % = (A 未加药 -A 加药 ) / (A 未加药 -A 空白 )×100%

[0058] In the formula, A 未加药A represents the absorbance value of the group without drug addition (containing cells and CCK-8 solution); 加药 represents the absorbance value of the drug-added group (containing cells, drugs and CCK-8 solution); A 空白 It represents the absorbance value of the blank group (only CCK-8 solution was added).

[0059] The effect of combined medication is evaluated according to King's formula: q value = E (a+b) / (E a +E b –E a ×E b ), E a+b is the tumor inhibition rate of the combination of the two drugs, E a 、E b is the tumor inhibition rate of the two drugs alone, q>1.15 indicates that the two drugs act synergistically; q<0.85 indicates that the two drugs antagonize each other; when q is 0.85-1.15, it indicates that the two drugs act additively.

[0060] 4. Animal Experiment Verification

[0061] Establishment of HepG2 liver cancer animal model: 18-20g BALB / c-nu male mice were adaptively fed for 5 days and each mouse was subcutaneously inoculated with HepG2 cells (2×10 7 ), and tumors can be seen growing after two weeks (50-100mm 3 The tumor-bearing mice were randomly divided into 4 groups: model group, psoralen group, icariin II group and combination group. After 14 days of continuous administration, the samples of the drug-treated groups were collected and statistical data were calculated: ① Tumor volume and weight: The length and width of the tumor of each group of mice were measured with a vernier caliper, and the formula (length × width) was used to calculate the tumor volume and weight. 2 ) / 2 to calculate the tumor volume; use a balance to weigh the tumor weight of each group of mice and record it. ② Tumor inhibition rate: calculate the average tumor weight of each group of mice (m1) and the average tumor weight of the model group mice (m 2 ), according to the formula (1-m1 / m 2 )×100% to obtain the tumor inhibition rate of each drug-treated group. ③ Organ coefficient: weigh the weight of each organ tissue such as the heart, liver, spleen, lung, and kidney of the mouse, and then divide it by the corresponding mouse body weight to calculate the percentage of each organ, that is, the organ coefficient. All experiments and procedures involving mice were performed in accordance with the guidelines of the Animal Ethics Committee of the Cancer Hospital of the Chinese Academy of Medical Sciences.

[0062] The measurement data were expressed as mean ± SD. One-way ANOVA was used for comparison among multiple groups, and t-test was used for comparison between two groups. p < 0.05 indicated a significant difference, and p < 0.01 indicated a very significant difference.

[0063] Example 2 Experimental results

[0064] like Figure 1A -K shows that the cytotoxicity of psoralen and psoralen in Psoralea corylifolia is dose-dependent within a certain concentration range, and the inhibition rate of psoralen is greater at low doses. Therefore, we chose psoralen for subsequent research.

[0065] like Figure 2A -E, the components of Epimedium have almost no effect on the growth of HepG2 cells at low concentrations. In the higher concentration range of Epimedium II, the inhibitory effect on the growth of HepG2 cells is stronger and in a dose-dependent manner.

[0066] Psoralen A (33.33 μM) at a non-toxic concentration was co-administered with 10 different concentrations of Epimedium monomers, and the cell inhibition rate and King's formula q value were determined. Figure 3A -H shows that icariin II has a synergistic inhibitory effect (q≥1.15) when it is administered at a dose of 8.33 and 16.67 μM in combination with psoralen; epimedin B and epimedin C have a synergistic inhibitory effect (q≥1.15) when they are administered at doses of 8.33, 16.67, 33.33 and 66.67 μM in combination with psoralen. In summary, icariin II, epimedin B and epimedin C at different concentrations combined with psoralen have a synergistic inhibitory effect on the growth of HepG2 cells. In addition, the King's formula q value is the largest when icariin II, epimedin B and epimedin C are combined with psoralen (33.33 μM) at 16.67 μM, 16.67 μM and 33.33 μM, respectively. Preferably, the optimal concentration ratios of psoralen A to icariin II, epimedin B and epimedin C are 2:1, 2:1 and 1:1, respectively.

[0067] In order to further screen the best combination, the compositions with different concentration gradients (psoralen + epimedin B, psoralen + epimedin C, psoralen + icariin II) were verified in vitro. Figure 4A -D, it was finally determined that the combination of psoralen A and icariin II had the strongest synergistic inhibitory effect on the growth of HepG2 cells (q≥1.15).

[0068] like Figure 5A -D shows that compared with psoralen or icariin II alone, the combination of psoralen and icariin II significantly reduced the tumor mass and volume of mice (p<0.01), and there was no significant change in body weight and organ coefficient. In vivo experiments further proved that the combination of psoralen and icariin II has a good anti-liver cancer effect.

Claims

1. Use of a composition for treating cancer in preparing a drug for treating liver cancer, characterized in that: The composition for treating cancer consists of psoralen A and icariin II in a weight ratio of 2:

1.

2. The use according to claim 1, wherein the drug is in an oral dosage form or a parenteral dosage form.

3. The use according to claim 2, wherein the drug is in an oral dosage form.

4. The use according to claim 1, wherein the medicine comprises a pharmaceutically acceptable carrier or excipient.

5. The use according to claim 4, wherein the pharmaceutically acceptable carrier or excipient is selected from one or more of a solvent, a cosolvent, a pH regulator, an osmotic pressure regulator, an antioxidant, a filler, a binder, a lubricant, and a disintegrant.

Images