CN111228319A

CN111228319A - Brucea javanica extract, preparation method thereof and application thereof in preventing and treating breast cancer

Info

Publication number: CN111228319A
Application number: CN202010234484.1A
Authority: CN
Inventors: 苏冀彦; 陈小红; 吕淑媚; 谢意珍
Original assignee: Guangdong Detection Center of Microbiology of Guangdong Institute of Microbiology
Current assignee: Guangdong Detection Center of Microbiology of Guangdong Institute of Microbiology
Priority date: 2020-03-30
Filing date: 2020-03-30
Publication date: 2020-06-05

Abstract

The invention relates to an application of a brucea javanica extract in preventing and/or treating breast cancer and a preparation method of the brucea javanica extract. The invention discloses an application of a brucea javanica extract in preventing and/or treating breast cancer. The invention discovers that the brucea javanica extract not only can play an anti-tumor role, but also has no obvious side effect on organisms, and has good application prospect.

Description

Brucea javanica extract, preparation method thereof and application thereof in preventing and treating breast cancer

Technical Field

The invention relates to an application of a brucea javanica extract in preventing and/or treating breast cancer and a preparation method of the brucea javanica extract.

Background

Cancer is a serious disease endangering human life health, wherein breast cancer is one of the most common malignant cancers of women and seriously threatens the life health of the women. Currently, methods for treating breast cancer can be divided into three categories, namely drug therapy (chemotherapy), radiotherapy and surgical treatment. The chemotherapy drugs used clinically mainly include: cyclophosphamide, methotrexate, 5-fluorouracil, adriamycin, taxol, etc. However, chemotherapy drugs often induce many adverse reactions in the application process, such as paclitaxel, and as a cytotoxic drug, due to lack of targeting specificity, clinical application of the drug can generate severe toxic and side effects such as myeloma inhibition, anaphylaxis, neurotoxicity, alopecia, drug-induced pain, and the like. Meanwhile, because the source of the chemotherapy drugs is limited and the price is high, a heavy economic burden is brought to patients. Therefore, in order to facilitate clinical use, reduce treatment cost and relieve pain of patients, finding substances which can effectively inhibit cancer development, reduce chemotherapeutic drug dosage and relieve adverse reactions becomes an urgent problem to be solved clinically.

Disclosure of Invention

In view of the defects, the brucea javanica extract can effectively prevent and/or treat breast cancer, has no obvious toxic or side effect on other organs including liver and kidney, and has a great prospect in the aspect of breast cancer treatment due to the obvious effect of inhibiting breast cancer cells.

The fructus Bruceae is dry mature fruit of Brucea Javanica (Linnaeus) Merrill (Brucea Javanica) belonging to Brucea (Simaroubaceae) of Simaroubaceae, also known as Brucea Javanica, Sophora flavescens and Brucea Javanica. Brucea javanica, originally recorded in Bencao gang mu Shi Yi, has cold bitter and small toxicity and enters large intestine meridian; has effects of clearing away heat and toxic materials, relieving dysentery, preventing malaria, and removing wart. The main components of the brucea javanica comprise picrasma quassioides lactone, alkaloid, fatty acid and the like, and the brucea javanica has wide pharmacological activity including anti-tumor, anti-inflammatory, antivirus, antibacterial, antiparasitic, blood sugar reducing and the like.

The brucea javanica oil is fatty oil extracted from brucea javanica fruits, and can be used for treating various malignant tumors, chronic gastritis, condyloma acuminatum and other diseases by penetrating a blood brain barrier, so that the brucea javanica oil is prepared into various dosage forms and becomes a well-known Chinese patent medicine to be sold on the market for many years. The preparation method of the brucea javanica oil is mature, and the common preparation method adopts petroleum ether as a solvent to extractBy ultrasonic extraction, Soxhlet extraction and supercritical CO₂Extraction method, etc.

However, the study finds that the brucea javanica also contains a large amount of quassinoid components, mainly including brucea javanica picrol, brucea javanica picrin and the like, and the components have good anti-tumor activity. Because the components are polar components, after the brucea javanica oil is extracted and prepared by the existing method, a large amount of brucea javanica oil still exists in medicine dregs basically, but the research on the medicine dregs is less at present, and the medicine dregs are not developed deeply and the comprehensive utilization of resources is not seen, so that the waste is caused.

The invention achieves the above purposes through the following scheme:

in a first aspect, the invention provides an application of a brucea javanica extract in preventing and/or treating breast cancer.

Wherein the brucea javanica extract comprises brucea javanica ethanol extract or brucea javanica oil.

In a second aspect, the invention provides an application of a brucea javanica extract in preparing a medicament for preventing and/or treating breast cancer.

In a third aspect, the invention provides a preparation method of an ethanol extract of brucea javanica, which comprises the following steps: adding ethanol solution into fructus Bruceae or fructus Bruceae residue after fatty acid extraction, continuously reflux-extracting, collecting reflux-extracted solution, repeating the above steps for 1-3 times to obtain precipitate, and mixing all filtrates; concentrating the filtrate under reduced pressure to obtain fructus Bruceae ethanol extract.

In the preparation method, the ethanol solution is added according to the feed-liquid ratio of 1: 4.

In the above production method, the ethanol solution is preferably a 95% ethanol aqueous solution.

In the above production method, the number of repetitions is preferably 2.

In the above steps, after vacuum concentration, the brucea javanica extract can be frozen and dried to obtain the brucea javanica ethanol extract.

The inventors have found that, in the above-mentioned production method, when a 95% aqueous ethanol solution is used and the number of repetitions is 2, a high extraction rate can be achieved, and for example, the extraction rate can be made 1.4% or more by using this method.

The inventors have found that the use of freeze-drying allows the maximum retention of the activity of the extract.

A method for preparing a preferred ethanol extract of brucea javanica comprises: taking brucea javanica or brucea javanica dregs after fatty acid extraction, adding 95% ethanol solution according to the liquid-material ratio of 1:4, carrying out continuous reflux extraction, collecting reflux extracting solution, repeating the steps for 2 times after the reflux extracting solution is left, and combining all filtrates; concentrating the filtrate under reduced pressure, and freeze drying to obtain ethanol extract of fructus Bruceae.

The preferable preparation method can obtain high extraction efficiency under the condition of most reasonable reagent and energy consumption, and ensure the inhibiting effect of the extract.

In a fourth aspect, the brucea javanica ethanol extract prepared by the preparation method is provided.

In a fifth aspect, a medicament for preventing and/or treating breast cancer is provided, which comprises an brucea javanica extract and a pharmaceutically acceptable carrier.

The oleum fructus brucease in the invention can be prepared by a conventional method or can be purchased in the market.

For example, one conventional method for preparing oleum fructus Bruceae: extracting fructus Bruceae with petroleum ether, distilling the extractive solution to recover petroleum ether, and filtering to obtain oleum fructus Bruceae.

The brucea javanica ethanol extract obtained by the preparation method is used for carrying out in-vitro anti-breast cancer cell experiments, and results show that the brucea javanica ethanol extract can obviously inhibit proliferation and autophagy of breast cancer cells after acting on the cells for 48 hours.

The brucea javanica ethanol extract obtained by the preparation method is used for carrying out in-vivo anti-tumor experiments on breast cancer mice, and the results show that the brucea javanica extract can obviously inhibit tumor growth, reduce tumor weight, reduce tumor volume and inhibit tumor tissue autophagy after gastric lavage, and has no obvious damage to small intestines, livers and kidneys.

Wherein, the ethanol extract IC of brucea javanica prepared by the invention₅₀The value is 10.42 mug/mL, and the inhibition effect is remarkable.

The brucea javanica oil obtained by the conventional method in the field is used for carrying out in-vivo anti-tumor experiments on breast cancer mice, and the results show that the brucea javanica oil can obviously inhibit tumor growth, reduce tumor weight, reduce tumor volume and inhibit autophagy of tumor tissues after gastric lavage, and has no obvious damage to small intestines, livers and kidneys.

In the application, after ethanol extraction is carried out on the brucea javanica or brucea javanica dregs after fatty acid extraction, the ethanol extract shows a very remarkable inhibition effect on the development of breast cancer, shows an anti-tumor effect close to that of the brucea javanica oil at a lower dose, and has no obvious toxic or side effect, so that the anti-tumor activity of the ethanol extract is better than that of the brucea javanica oil; on the other hand, the ethanol extract obtained from the brucea javanica or the brucea javanica dregs after the fatty acid extraction has no obvious difference in anti-tumor effect, which indicates that the brucea javanica oil and the ethanol extract of the brucea javanica or the brucea javanica dregs after the fatty acid extraction are active ingredients of brucea javanica for resisting the breast cancer.

Compared with the prior art, according to the preparation method disclosed by the invention, brucea javanica seeds or dregs of brucea javanica seeds after fatty acid extraction are used as raw materials, ethanol is used as an extraction solvent, and the preparation is completed through steps of heating reflux extraction, reduced pressure concentration, freeze drying and the like.

The inventor further compares the inhibition effect of the brucea javanica ethanol extract and the brucea javanica oil on the breast cancer cells respectively, and finds that the brucea javanica ethanol extract can obviously inhibit the growth of the breast cancer cells in vitro and the brucea javanica oil has no obvious influence on the breast cancer cells; in vivo, the brucea javanica oil and the brucea javanica ethanol extract can inhibit the growth of breast cancer cells, and the effects of the brucea javanica oil and the brucea javanica ethanol extract are similar. This may suggest that ethanol extract of brucea javanica has a better potential for treating breast cancer.

Compared with the prior art, the invention has the following advantages:

in the extraction process of the brucea javanica ethanol extract, the brucea javanica medicinal material is fully utilized, ethanol is used as a solvent, and an extraction method of heating reflux extraction, reduced pressure concentration and freeze drying is adopted, so that the brucea javanica ethanol extract has the advantages of being environment-friendly in reagent, simple in process, low in production cost and capable of effectively avoiding organic solvent pollution, and the extraction method can retain the activity of the brucea javanica extract to the maximum extent, and IC (integrated circuit) of the brucea javanica ethanol extract₅₀The value is low, and the extracted brucea javanica extract has the prospect of preparing medicaments for treating breast cancer.

The invention discovers that the brucea javanica extract not only can play an anti-tumor role, but also has no obvious side effect on organisms, and has good application prospect.

Drawings

The brucea javanica extract, the extraction method and the application thereof and the beneficial effects are described in detail below with reference to the accompanying drawings and the specific embodiments.

FIG. 1 is a graph showing the results of measuring the components of ethanol extract of brucea javanica according to example 1 of the present invention.

FIG. 2 shows the effect of ethanol extract of brucea javanica on the viability of breast cancer cells in test example 1 of the present invention.

FIG. 3 is a western blot showing the expression level of autophagy-related protein in each group of cells in Experimental example 1 of the present invention.

FIG. 4 is a graph showing the change in the expression level of ULK1 protein in each group of cells in Experimental example 1 of the present invention.

FIG. 5 is a graph showing the change in the expression level of Beclin-1 protein in each group of cells in Experimental example 1 of the present invention.

FIG. 6 is a graph showing the change in the expression level of p62 protein in each group of cells in Experimental example 1 of the present invention.

FIG. 7 is a graph showing changes in the expression level of LC3 II/I protein in each group of cells in Experimental example 1 of the present invention.

FIG. 8 is a graph showing the change in tumor tissue volume in each group of animals in Experimental example 2 of the present invention.

FIG. 9 is a graph showing the change in tumor tissue weight in each group of animals in Experimental example 2 of the present invention.

FIG. 10 is a graph showing a comparison of HE staining of small intestinal tissues in each group of animals in Experimental example 2 of the present invention.

FIG. 11 is a graph showing a comparison of HE staining of liver tissues of each group of animals in Experimental example 2 of the present invention.

FIG. 12 is a graph showing a comparison of HE staining of kidney tissue in each group of animals in Experimental example 2 of the present invention.

FIG. 13 is a western blot of autophagy-associated proteins in tumor tissues of various groups of animals in Experimental example 2 of the present invention.

FIG. 14 is a graph showing the change in the expression level of ULK1 protein in tumor tissue of each group of animals in Experimental example 2 of the present invention.

FIG. 15 is a graph showing the change in the expression level of Beclin-1 protein in tumor tissue of each group of animals in Experimental example 2 of the present invention.

FIG. 16 is a graph showing the change in the expression level of p62 protein in tumor tissue of each group of animals in Experimental example 2 of the present invention.

FIG. 17 is a graph showing the change in the expression level of LC3 II/I protein in tumor tissue of each group of animals in Experimental example 2 of the present invention.

FIG. 18 is a graph showing the change in tumor tissue volume in each group of animals in Experimental example 3 of the present invention.

FIG. 19 is a graph showing the change in tumor tissue weight in each group of animals in Experimental example 3 of the present invention.

FIG. 20 is a graph showing a comparison of HE staining of small intestinal tissues in each group of animals in Experimental example 3 of the present invention.

FIG. 21 is a graph showing a comparison of HE staining of liver tissues of each group of animals in Experimental example 3 of the present invention.

FIG. 22 is a graph showing a comparison of HE staining of kidney tissue in each group of animals in Experimental example 3 of the present invention.

FIG. 23 is a western blot of autophagy-associated proteins in tumor tissues of various groups of animals in Experimental example 3 of the present invention.

FIG. 24 is a graph showing the change in the expression level of ULK1 protein in tumor tissue of each group of animals in Experimental example 3 of the present invention.

FIG. 25 is a graph showing the change in the expression level of Beclin-1 protein in tumor tissue of each group of animals in Experimental example 3 of the present invention.

FIG. 26 is a graph showing the change in the expression level of p62 protein in tumor tissue of each group of animals in Experimental example 3 of the present invention.

FIG. 27 is a graph showing the change in the expression level of LC3 II/I protein in tumor tissue of each group of animals in Experimental example 3 of the present invention.

Detailed Description

In order to make the objects, technical solutions and advantageous technical effects of the present invention clearer, the present invention is further described in detail with reference to the following embodiments. It should be understood that the embodiments described in this specification are only for the purpose of illustrating the invention and are not to be construed as limiting the invention, and the parameters, proportions and the like of the embodiments may be suitably selected without materially affecting the results.

The reagents, methods and apparatus employed in the examples of the invention are conventional in the art, unless otherwise indicated.

The brucea javanica seeds comprise brucea javanica seeds.

Example 1 preparation of brucea javanica pomace

Brucea javanica, purchased from Baiyunshanmingxing pharmaceuticals, Inc.

Adding petroleum ether as an extraction solvent into brucea javanica according to the ratio of the material to the liquid being 1:5, carrying out continuous heating reflux extraction, filtering, collecting filter residues and drying to obtain brucea javanica dregs.

Example 2 preparation and composition analysis of ethanol extract of brucea javanica

Brucea javanica, purchased from Baiyunshanmingxing pharmaceuticals, Inc.

Taking brucea javanica or brucea javanica dregs after fatty acid extraction, adding 95% ethanol according to the ratio of material to liquid of 1:4, carrying out continuous reflux extraction, filtering, collecting filtrate, repeating the extraction process twice on the left precipitate, combining the filtrates obtained in three times, concentrating under reduced pressure until no ethanol smell exists, and carrying out freeze drying to obtain the brucea javanica ethanol extract with the extraction rate of 1.4%.

Analyzing the components of the brucea javanica extract: dissolving brucea javanica ethanol extract in methanol, using water and methanol as mobile phase, and performing component detection by liquid chromatography (HPLC) with detection wavelength of 240nm, wherein the chromatogram of brucea javanica extract is shown in figure 1.

As can be seen from FIG. 1, the ethanol extract of brucea javanica was detected to contain a number of components at 240nm, including brucea javanica alcohol (retention time about 33min) as seen by comparison of retention times.

Test example 1 inhibitory Effect of ethanol extract of brucea javanica on Breast cancer cells

1. Effect of ethanol extract of brucea javanica on MDA-MB-231 cell viability

Culturing breast cancer cells (MDA-MB-231) in DEME complete medium (containing 1% of streptomycin double antibody and 10% of fetal calf serum), maintaining the temperature at 37 deg.C and 5% of CO₂Culturing in a sterile incubator with saturated humidity. Collecting cells in logarithmic growth phase, washing with phosphate buffer salt twice, adding appropriate amount of pancreatin to digest cells, collecting cells, centrifuging, adding appropriate amount of above culture medium, resuspending, and adjusting cell concentration to 3 × 10⁴one/mL, then seeded in 96-well plates. After overnight incubation, brucea javanica ethanol extract solution (prepared as in example 2) was added to a final concentration of 0.78-200. mu.g/mL. After 48h of culture, 20. mu.L of thiazole blue solution (MTT) was added to each well, and after 4h of culture, the OD value of the optical density was measured at 490nm of a microplate reader, and the respective IC's were calculated₅₀Values versus percent cell viability.

2. Detection of cell autophagy-related protein expression level by Western Blot (WB)

Taking cells in logarithmic growth phase, adjusting cell concentration to 1 × 10⁵one/mL, seeded in 6-well plates. After overnight incubation, 5 groups were set up as shown in table 1. After 48h of culture, 6-well plate cells were harvested and assayed for expression levels of autophagy-related proteins (ULK1, p62, LC3 II/I) using WB technology. Wherein "+". X "represents p, respectively, as compared with the blank control group<0.05，p<0.01。

TABLE 1

3. Results

As shown in figure 2, after 48 hours of administration, brucea javanica ethanol extract can obviously inhibit MDA-MB-231 cell activity and has certain concentration dependence (1.5625-200 mu g/mL) of IC₅₀The value is 10.42 mu g/mL, which shows that the brucea javanica ethanol extract can effectively inhibit the growth of tumor cells, has the anti-tumor effect in vitro and has the prospect of preparing medicaments for treating breast cancer.

The test results in fig. 3 to 7 show that, compared with the blank control group, the expression levels of ULK1 and Beclin-1 protein in the brucea javanica ethanol extract group (medium concentration and high concentration group) are obviously reduced, while the expression level of p62 protein is increased (low concentration and medium concentration group), and the expression ratio of LC3 II/I protein is obviously reduced (high concentration group). The brucea javanica ethanol extract can inhibit MDA-MB-231 autophagy, thereby playing an anti-tumor role.

Test example 2 inhibitory Effect of ethanol extract of brucea javanica on Breast cancer in tumor-bearing mice

1. Modeling and administration

Taking Balb/c nude mice and females. After 3 days of acclimatization, the animals were randomly divided into a normal group (10) and a tumor-bearing group (40) by body weight. Tumor-bearing group inoculated breast cancer cell MDA-MB-231 (2X 10)⁶One/one), a mouse breast cancer model was established. When the tumor volume grows to 1mm³Tumor-bearing groups were randomly divided, and administered for 21 consecutive days as shown in Table 2, and the normal group and the model group were administered with an equal volume of physiological saline.

TABLE 2

2. Taking materials and detecting

During the experiment, tumor volumes were measured twice weekly. After the last administration for 24h, mice were dislocated and sacrificed, and tumors, small intestine, liver, and kidney were dissected out. Tumors were photographed, weighed and frozen for immunoblotting (WB) to determine the expression level of autophagy-related proteins (ULK1, p62, Beclin-1, LC3 II/I). Tissues of small intestine, liver and kidney are taken and used for hematoxylin-eosin staining (HE) to observe tissue change. Wherein "" + ". indicates p <0.05 and p <0.01, respectively, as compared to the model group.

3. Results

As shown in fig. 8 and fig. 9, after 21 days of administration, the paclitaxel group and the brucea javanica ethanol extract group (high and low dose groups) can both obviously inhibit the growth of tumors, and the volume and the weight of the tumors are obviously reduced. The brucea javanica ethanol extract has obvious anti-tumor effect in tumor-bearing mice.

The HE staining results of fig. 10, 11, 12 show that paclitaxel group has altered small intestine tissue structure compared with the model group, mainly concentrated in the side muscle layer loose, and the intestinal villi is obviously shortened; the liver tissue has obvious inflammatory infiltration, and compared with the taxol group, the small intestine of the brucea javanica ethanol extract group (high and low dose groups) has no obvious injury, and the liver tissue has no inflammatory infiltration. The results show that the anti-tumor effect of the brucea javanica ethanol extract can not cause the toxic and side effects of non-target organs.

The test results in fig. 13 to fig. 17 show that the expression levels of ULK1 and Beclin-1 proteins are obviously reduced in the brucea javanica ethanol extract high-dose group, and the ratio of LC3 II/I proteins is also obviously reduced compared with the model group. The results show that the brucea javanica ethanol extract can play an anti-tumor role by inhibiting autophagy of cells in a tumor-bearing mouse body.

Test example 3 inhibitory Effect of oleum fructus Bruceae on Breast cancer in tumor-bearing mice

1. Materials and methods

Female Balb/c nude mice of 4-5 weeks old were purchased from the center of laboratory animals (animal license number: GT-IACUC201807262) in Guangdong province; paclitaxel injection (100mg/16.7mL), supplied by Hainan Quanxing pharmaceutical Co., Ltd; brucea javanica oil emulsion (cat # 160202) was purchased from Mingxing pharmaceuticals, Inc. of Baiyunshan, Guangzhou and contained 10% brucea javanica oil.

2. Modeling and administration

Taking Balb/c nude mice and femalesAnd (4) sex. After 3 days of acclimatization, the animals were randomly divided into a normal group (10) and a tumor-bearing group (50) by body weight. Tumor-bearing group inoculated breast cancer cell MDA-MB-231 (2X 10)⁶One/one), a mouse breast cancer model was established. When the tumor volume grows to 1mm³Tumor-bearing groups were randomly divided, and administered for 21 consecutive days as shown in Table 3, and the normal group and the model group were administered with an equal volume of physiological saline.

TABLE 3

3. Taking materials and detecting

4. Results

As shown in FIGS. 18 and 19, after 21 days of administration, paclitaxel and oleum fructus Bruceae (high, medium and low dose groups) both significantly inhibited the growth of tumor tissues, resulting in significant reduction in the volume and weight of tumor tissues. The brucea javanica oil has obvious anti-tumor effect in tumor-bearing mice, and the effect is similar to the anti-tumor effect of the brucea javanica ethanol extract (see the figure 8 and the figure 9).

The HE staining results of fig. 20, 21, 22 show that paclitaxel group has altered small intestine tissue structure compared with the model group, mainly concentrated in the side muscle layer loose, and the intestinal villi is obviously shortened; the liver tissue has obvious inflammatory infiltration, and compared with taxol, the small intestine of the brucea javanica oil group (high, medium and low dose groups) has no obvious injury, and the liver tissue has no inflammatory infiltration. The results show that the antitumor effect of the oleum fructus bruceae can not cause the toxic and side effects of non-target organs.

The test results in fig. 23 to 27 demonstrate that compared with the model group, the expression level of ULK1 protein and the ratio of LC3 II/I protein expression are significantly reduced in the brucea javanica oil group (medium dose group, high dose group), and the expression level of Beclin-1 protein is significantly reduced (high dose group). The brucea javanica oil can inhibit autophagy of cells in a tumor-bearing mouse body, thereby playing an anti-tumor role.

Appropriate changes and modifications to the embodiments described above will become apparent to those skilled in the art from the disclosure and teachings of the foregoing description. Therefore, the present invention is not limited to the specific embodiments disclosed and described above, and some modifications and variations of the present invention should fall within the scope of the claims of the present invention. Furthermore, although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.

Claims

1. Application of fructus Bruceae extract in preparing medicine for preventing and/or treating breast cancer is provided.

2. The use as claimed in claim 1, wherein the brucea javanica extract comprises an ethanol extract of brucea javanica or brucea javanica oil.

3. The preparation method of the brucea javanica ethanol extract is characterized by comprising the following steps: adding ethanol solution into fructus Bruceae or fructus Bruceae residue after fatty acid extraction, continuously reflux-extracting, collecting reflux-extracted solution, repeating the above steps for 1-3 times to obtain precipitate, and mixing all filtrates; concentrating the filtrate under reduced pressure to obtain fructus Bruceae ethanol extract.

4. The preparation method according to claim 3, characterized in that the ethanol solution is added in a feed-to-liquid ratio of 1: 4; or the ethanol solution is 95% ethanol water solution.

5. The method according to claim 3, wherein the number of repetitions is preferably 2.

6. The process according to claim 3, wherein the ethanol extract of brucea javanica is obtained by freeze-drying after concentration under reduced pressure.

7. The method according to claim 3, wherein a 95% ethanol aqueous solution is used, and the number of repetitions is 2.

8. The method of claim 3, comprising: taking brucea javanica or brucea javanica dregs after fatty acid extraction, adding 95% ethanol solution according to the liquid-material ratio of 1:4, carrying out continuous reflux extraction, collecting reflux extracting solution, repeating the steps for 2 times after the reflux extracting solution is left, and combining all filtrates; concentrating the filtrate under reduced pressure, and freeze drying to obtain ethanol extract of fructus Bruceae.

9. A medicine for preventing and/or treating breast cancer is characterized by comprising brucea javanica extract and a pharmaceutically acceptable carrier.

10. The medicament of claim 9, wherein the brucea javanica extract comprises an ethanol extract of brucea javanica or brucea javanica oil.