Traditional Chinese medicine composition for treating triple-negative breast cancer pulmonary metastasis and preparation method and application thereof

Abstract

The invention discloses a traditional Chinese medicine composition for treating triple-negative breast cancer pulmonary metastasis, which is prepared from the following raw material medicines in parts by weight: 3-5 parts of herba epimedii, 3-5 parts of pseudobulbus cremastrae seu pleiones, 5-15 parts of rhizoma curcumae, 2-8 parts of nidus vespae and 5-15 parts of fructus akebiae. The traditional Chinese medicine composition disclosed by the invention has the effects of removing blood stasis and removing stasis, soothing liver and regulating qi, tonifying kidney and strengthening bone, and removing stasis and detoxifying, and can improve the disease-free survival rate of a metastatic breast cancer patient after operation for 2 years, delay the recurrence and metastasis of the postoperative breast cancer, enhance the immunity of the patient and improve the clinical symptoms of the patient. In addition, the invention further discloses a preparation method and application of the traditional Chinese medicine composition.

Description

Technical Field

[0001] The present invention relates to a traditional Chinese medicine composition for treating lung metastasis of triple-negative breast cancer. In addition, the present invention relates to a preparation method and application of the traditional Chinese medicine composition. Background Art

[0002] Breast cancer is currently one of the leading causes of cancer-related deaths in women. Recent studies have found that breast cancer accounts for 11.7% of all tumors and 30% of all cancer cases in women. Among them, triple-negative breast cancers (TNBC) account for about 15%-20% of all invasive breast cancers. Because it lacks estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER-2), it is insensitive to estrogen receptor antagonists, aromatase inhibitors, trastuzumab and pertuzumab. The recommended treatment options for TNBC are only single-drug sequential chemotherapy or combined chemotherapy. The limitations of the treatment method make it more prone to recurrence, metastasis and chemotherapy resistance. More than 90% of deaths are caused by recurrence and metastasis. Common metastatic lesions are mostly found in the lungs, bones, liver and brain. Studies have shown that 58.0% of TNBC have lung metastases, which are more dangerous. Literature reports that the 5-year survival rate of patients with TNBC of III-IV is only 13%. At present, although surgery, radiotherapy, chemotherapy and targeted therapy represented by EGFR inhibitors can improve clinical symptoms to a certain extent, they are prone to short-term recurrence risks, chemotherapy resistance and serious adverse reactions (gastrointestinal reactions, bone marrow function suppression, hair loss, etc.). Traditional Chinese medicine is increasingly showing its advantages in the adjuvant treatment of inhibiting metastasis of TNBC patients. It can cooperate with surgery, radiotherapy, chemotherapy and other treatment methods to regulate the body's immune function, reduce the toxicity of radiotherapy and chemotherapy, prevent recurrence and metastasis, and prolong the survival of tumor-bearing patients.

[0003] At present, the main methods of TCM-assisted treatment of metastasis in TNBC patients in clinical practice are dialectical treatment and prescription preparation. There is no effective Chinese medicine combination in the form of preparation to benefit patients. Summary of the invention

[0004] In view of the above-mentioned deficiencies in the prior art, according to an embodiment of the present invention, it is hoped to provide a Chinese medicine composition for treating TNBC lung metastasis, and propose a preparation method and application thereof.

[0005] In a first aspect, the present invention provides a traditional Chinese medicine composition for treating TNBC lung metastasis, which is prepared from the following raw materials in parts by weight: 3-5 parts of Epimedium, 3-5 parts of Trillium, 5-15 parts of Curcuma, 2-8 parts of Apium graveolens, and 5-15 parts of Rhizoma Atractylodis Macrocephalae.

[0006] Preferably, the traditional Chinese medicine composition for treating TNBC lung metastasis is prepared from the following raw materials in parts by weight: 4 parts of Epimedium, 4 parts of Trillium, 10 parts of Curcuma, 5 parts of Apium graveolens, and 10 parts of Rhizoma Atractylodis Macrocephalae.

[0007] In a second aspect, the present invention provides a method for preparing a traditional Chinese medicine composition for treating TNBC lung metastasis, comprising the following steps: weighing the raw materials by weight: epimedium, trillium, zedoaria, beehive and aconite, adding water and decocting twice, adding 8-12 times the amount of water for the first time, decocting for 2-4 hours, adding 8-12 times the amount of water for the second time, decocting for 1-1.5 hours, combining the decoctions, filtering, and concentrating the filtrate to a relative density of 1.14 to 1.15 (60-80°C).

[0008] Preferably, after the concentration and extraction step of the traditional Chinese medicine composition, the step further includes: adding pharmaceutically acceptable excipients to prepare granules.

[0009] In a third aspect, the present invention provides use of the aforementioned Chinese medicine composition in the preparation of a drug for treating TNBC lung metastasis.

[0010] The pharmacological analysis of the Chinese medicine composition for treating TNBC lung metastasis of the present invention is as follows:

[0011] Epimedium: pungent, sweet, warm. Enters the liver and kidney meridians. It has the effects of tonifying the kidney and strengthening yang, strengthening the tendons and bones, and removing rheumatism. It can enhance the secretion function of the endocrine system.

[0012] Trillium: sweet, slightly pungent, cold. Enters the liver, stomach, and spleen meridians. It has the effects of clearing away heat and detoxifying, resolving phlegm and dispersing nodules, and can play a variety of pharmacological effects such as anti-tumor, antibacterial, and lipid-lowering.

[0013] Curcuma: pungent, bitter, warm. Enters the liver and spleen meridians. Has the effects of breaking blood and promoting qi, eliminating accumulation and relieving pain. It is often used for lumps, blood stasis, amenorrhea, chest pain, etc.

[0014] Beehive: sweet in taste, neutral in nature. Enters the liver, stomach, and kidney meridians. Has the effects of dispelling wind and relieving pain, attacking poison and killing insects, killing insects and relieving itching, and drying dampness and detoxifying. It is mostly used to treat sores, swellings, breast abscesses, scrofula, stubborn skin ringworm, etc.

[0015] August Zha: bitter, cold in nature. Enters the liver and stomach meridians. Has the effects of soothing the liver and regulating qi, promoting blood circulation and relieving pain, and relieving restlessness and promoting diuresis. Can enhance the body's immunity, anti-inflammatory, and antibacterial.

[0016] The traditional Chinese medicine composition for treating TNBC lung metastasis of the present invention uses Trillium gracile and Apis cerana as the main medicines, both of which have strong anti-cancer and nodule-dispersing effects and have the meaning of "fighting poison with poison", and Apis cerana also has the effect of warming the kidney; Curcuma zedoaria is used as the assistant medicine to break up blood stasis and attack accumulation, and has the effect of eliminating tumors and dispersing nodules; and Epimedium and Achyranthes bidentata are used as the adjuvants to soothe the liver and regulate qi, nourish the kidney and strengthen the bones, and make the whole prescription eliminate evil without hurting the body, so as to better play the role of dispersing nodules and detoxifying.

[0017] Subsequent test examples will demonstrate that the Chinese medicine composition of the present invention has potential advantages in the treatment of metastatic breast cancer; has a significant anti-tumor growth effect on TNBC lung metastasis mice; has a significant inhibitory effect on 4T1 and MDA-MB-231 breast cancer cells; can reduce the invasive ability of TNBC cells; and has a significant therapeutic effect on inhibiting breast cancer lung metastasis. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The effect of the Chinese medicine composition of the present invention on the tumor volume and weight of mice with breast cancer lung metastasis [tumor volume abc / 2. Model: model control group (inoculated with 4T1 cells); Chinese medicine composition treatment group of the present invention (low, medium and high dose groups of the Chinese medicine composition of the present invention were inoculated with 4T1 cells + 0.85g / kg / d, 1.7g / kg / d, and 2.55g / kg / d of the Chinese medicine composition of the present invention, respectively); Taxol: positive drug treatment group (inoculated with 4T1 cells + 0.01g / kg / week). (Comparison of tumor size between different groups)]

[0019] Figure 2 The effect of the Chinese medicine composition of the present invention on lung metastasis of breast cancer lung metastasis mice is shown.

[0020] Figure 3 The effect of the Chinese medicine composition of the present invention on the proliferation ability of TNBC cells is shown in ** P<0.01vs Controlgroup (0μg / mL; 24h, 48h, 72h)】

[0021] Figure 4 The effect of the Chinese medicine composition of the present invention on the migration ability of TNBC cells ** P<0.01vs Control group (0μg / mL; 24h, 48h); A, C: Experimental results and statistical graphs of the wound healing experiment of MDA-MB-231 cells intervened by the Chinese medicine composition of the present invention for 24h and 48h; B, D: Experimental results and statistical graphs of the wound healing experiment of 4T1 cells intervened by the Chinese medicine composition of the present invention for 24h and 48h]

[0022] Figure 5 The effect of the Chinese medicine composition of the present invention on the invasion ability of TNBC cells is shown in* P<0.05, ** P<0.01vsControl group (0μg / mL; 24h, 48h); A, C: experimental results and statistical graphs of the 24h, 48h Transwell chamber experiment of MDA-MB-231 cells intervened by the Chinese medicine composition of the present invention; B, D: experimental results and statistical graphs of the 24h, 48h Transwell chamber experiment of 4T1 cells intervened by the Chinese medicine composition of the present invention].

[0023] Figure 6 The base peak chromatogram (BPC) of the Chinese medicine composition of the present invention obtained by LC-mass spectrometry analysis [A: BPC graph in positive ion mode; B: BPC graph in negative ion mode].

[0024] Figure 7 The mass spectrum of the blood-entering components of the Chinese medicine composition of the present invention is shown as follows [Icariin; B. Xanthine; C. Xanthine acid; D. Guanosine].

[0025] Figure 8 Showing the mass spectrum of the lung components of the Chinese medicine composition of the present invention [A. Icariin; B. Icariin II]. DETAILED DESCRIPTION

[0026] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments. These embodiments should be understood to be only used to illustrate the present invention and not to limit the scope of protection of the present invention. After reading the contents of the present invention, those skilled in the art may make various changes or modifications to the present invention, and these equivalent changes and modifications also fall within the scope defined by the claims of the present invention.

[0027] Example 1

[0028] Weigh 40 grams of Epimedium, 40 grams of Trillium, 100 grams of Curcuma, 50 grams of Beehive, and 100 grams of Atractylodes, add water to each raw material and boil twice, filter and combine the filtrate, concentrate under reduced pressure to form an extract, dry at 70℃-80℃, crush and sieve, add appropriate amount of dextrin, mix well, dry, and make granules.

[0029] Example 2

[0030] Weigh 30 grams of Epimedium, 30 grams of Trillium, 100 grams of Curcuma, 50 grams of Beehive, and 100 grams of Atractylodes, add water to each raw material and boil twice, filter and combine the filtrate, concentrate under reduced pressure to form an extract, dry at 70℃-80℃, crush and sieve, add appropriate amount of dextrin, mix well, dry, and make granules.

[0031] Example 3

[0032] Weigh 40 grams of Epimedium, 50 grams of Trillium, 100 grams of Curcuma, 50 grams of Beehive, and 150 grams of Atractylodes, add water to each raw material and boil twice, filter and combine the filtrate, concentrate under reduced pressure to form an extract, dry at 70℃-80℃, crush and sieve, add appropriate amount of dextrin, mix well, dry, and make granules.

[0033] Example 4

[0034] Weigh 40 grams of Epimedium, 40 grams of Trillium, 100 grams of Curcuma, 50 grams of Beehive, and 150 grams of Atractylodes, add water to each raw material and boil twice, filter and combine the filtrate, concentrate under reduced pressure to form an extract, dry at 70℃-80℃, crush and sieve, add appropriate amount of dextrin, mix well, dry, and make granules.

[0035] Example 5

[0036] Weigh 40 grams of Epimedium, 40 grams of Trillium, 100 grams of Curcuma, 100 grams of Beehive, and 50 grams of Achyranthes bidentata, add water to each raw material and boil twice, filter and combine the filtrate, concentrate under reduced pressure to form an extract, dry at 70℃-80℃, crush and sieve, add appropriate amount of dextrin, mix well, dry, and make granules.

[0037] Example 6

[0038] Weigh 40 grams of Epimedium, 40 grams of Trillium, 100 grams of Curcuma, 50 grams of Beehive, and 100 grams of Atractylodes, add water to each raw material and boil twice, filter and combine the filtrate, concentrate under reduced pressure to form an extract, dry at 70℃-80℃, crush and sieve, add appropriate amount of dextrin, mix well, dry, and make granules.

[0039] Test Example 1-Clinical Efficacy Test

[0040] 1. Experimental drugs

[0041] During the preparation process of the Chinese medicine composition described in Example 6, the decoctions are combined and concentrated under reduced pressure to obtain an extract for use in clinical pharmacodynamic experiments.

[0042] 2. Observation of drug efficacy

[0043] Breast cancer patients whose first surgery time was between May 1, 2019 and May 31, 2022 were screened from the outpatient database of Professor Liu Sheng of Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine. Specific inclusion criteria: ① After surgical treatment of primary breast cancer, pathological examination confirmed that it was a malignant epithelial breast tumor (invasive breast cancer); ② The time of primary surgery was between May 1, 2019 and May 31, 2022; ③ Patients with clinical stage I-III who were operable and metastasis was found by postoperative pathology; ④ Female patients aged 18 to 90 years; ⑤ Those who voluntarily joined this trial to take Chinese medicine and continued to take the Chinese medicine composition of the present invention for ≥6 months. Observe the 2-year disease-free survival (DFS) of patients: Calculate the ratio of the number of cases without disease-related recurrence or metastasis to the total number of cases within 2 years since the patient received surgical treatment.

[0044] The results of the study showed that the two-year DFS of the medication group reached 89.80% (Table 1), and the two-year DFS of patients with lymph node metastasis alone reached 95.23%. The existing literature reported that the two-year DFS of patients with non-metastatic breast cancer was 83.50%, and the efficacy was significantly lower than that of patients with metastatic breast cancer in this study. This finding suggests that the Chinese medicine composition of the present invention has potential advantages in the treatment of metastatic breast cancer.

[0045] Table 1. 2-year DFS of patients with metastatic breast cancer taking the Chinese medicine composition of the present invention

[0046]

[0047] △ indicates that only local infiltration, lymph node metastasis or distant metastasis occurs;

[0048] ▲Indicates patients who achieved pCR with preoperative neoadjuvant chemotherapy and did not receive postoperative adjuvant chemotherapy.

[0049] Test Example 2 - In vivo experimental efficacy observation [Effect of the Chinese medicine composition of the present invention on tumor volume and weight of mice with breast cancer lung metastasis]

[0050] Female Balb / c mice aged 6-8 weeks were selected and injected with 4T1 cell suspension into the left fat pad of the second mammary gland at a cell density of 2×10 5 / mL, 0.1mL each, to establish a TNBC lung metastasis mouse model. All modeling animals were randomly divided into: normal group, model group, low-dose group of the Chinese medicine composition of the present invention (0.85g / kg), medium-dose group of the Chinese medicine composition prepared in Example 6 of the present invention (1.7g / kg), high-dose group of the Chinese medicine composition prepared in Example 6 of the present invention (2.55g / kg), paclitaxel-positive drug group (0.01g / kg). Each group began to be administered 14 days after inoculation of 4T1 cells. The low, medium and high-dose groups of the Chinese medicine composition of the present invention were gavaged and administered continuously for 21 days, once a day; the paclitaxel-positive drug group was 0.01g / kg, intraperitoneally injected, once a week. The normal group and the control group were gavaged with equal volumes of distilled water, once a day, for 21 consecutive days. During the treatment period, the growth status of the mice was observed, and the changes in tumor volume and mouse weight were recorded.

[0051] The measurement results show ( Figure 1 ), compared with the model group, the group treated with the Chinese medicine composition of the present invention was able to significantly inhibit the volume of breast cancer mice tumors (P < 0.01); picric acid soaked mouse lung tissue ( Figure 2 ), it can be found that compared with the model group, the group treated with the Chinese medicine composition of the present invention can significantly inhibit lung metastasis of breast cancer mice (P < 0.01).

[0052] In summary, the Chinese medicine composition of the present invention has a significant anti-tumor growth effect on TNBC lung metastasis mice and inhibits lung metastasis in breast cancer mice.

[0053] Test Example 3-In vitro experimental efficacy observation

[0054] 1. Drug preparation

[0055] The Chinese medicine composition described in Example 6 is prepared by heating Trillium gracile with ethanol and reflux twice and then mixing the filtrates; heating Curcuma zedoaria, Nymphaeaceae, Epimedium brevicornum and Rhizoma Atractylodis Macrocephalae with water and reflux twice and then mixing the filtrates; then heating the five medicinal residues with ethanol and reflux for the third time; and all the medicinal liquids are mixed, filtered, evaporated and dried.

[0056] 2. Pharmacodynamic observation

[0057] (1) Effect of the Chinese medicine composition of the present invention on the proliferation ability of TNBC cells.

[0058] Take TNBC cells MDA-MB-231 and 4T1 in the logarithmic growth phase and adjust the cell density to 5×10 4 cells / mL, inoculated in a 96-well cell culture plate (100 μL / well), with 6 replicates in each group, a blank control group and different concentration groups of the Chinese medicine composition of the present invention (0, 12.5, 50, 100 and 200 μg / ml) were set up, and after 24, 48, and 72 hours of intervention, the CCK8 method was used to detect the changes in the proliferation ability of TNBC cells after drug intervention.

[0059] The results showed that compared with the blank control group, different concentrations of the Chinese medicine composition of the present invention could significantly reduce the survival rate of MDA-MB-231 and 4T1 cells after 24, 48, and 72 hours of intervention (P<0.05 or P<0.01), and there was a dose-dependency, indicating that the drug can inhibit the proliferation ability of two TNBC cells (such as Figure 3 , as shown in Table 2-3).

[0060] Table 2. Effect of the Chinese medicine composition of the present invention on the proliferation ability of 4T1 cells

[0061]

[0062] ** P<0.01vs Control group(0μg / mL; 24h, 48h, 72h)

[0063] Table 3. Effect of the Chinese medicine composition of the present invention on the proliferation ability of MDA-MB-231 cells

[0064]

[0065] ** P<0.01vs Control group(0μg / mL; 24h, 48h, 72h)

[0066] (2) Effect of the Chinese medicine composition of the present invention on the migration ability of TNBC cells.

[0067] Human breast cancer cells MDA-MB-231 and mouse breast cancer cells 4T1 in the logarithmic growth phase were inoculated into 6-well cell culture plates for cell scratching, and basal culture medium or 25, 50, and 100 μg / mL of the Chinese medicine composition of the present invention were added and cultured for 24 h and 48 h, respectively. Pictures were taken under an inverted microscope, and the healing area and migration rate of the scratch area were calculated.

[0068] The results showed that compared with the blank control group, the Chinese medicine composition of the present invention at 25, 50, and 100 μg / mL could significantly reduce the migration rate of MDA-MB-231 and 4T1 cells after intervention for 24 h and 48 h (P<0.01, Figure 4, Table 4-5). For MDA-MB-231 cells, the cell migration rate was 1.8%-13.6% after 24h intervention with 100 μg / mL of the Chinese medicine composition of the present invention, and the cell migration rate was 8.8%-12.8% after 48h intervention, which was also statistically significant compared with the control group (P<0.01). For 4T1 cells, after 24h intervention with the Chinese medicine composition of the present invention, the cell migration rate at concentrations of 25, 50, and 100 μg / mL was only 5.6%-10.5%, which was statistically significant compared with the control group (P<0.01); after 48h intervention, compared with the blank control group, the cell migration rate at concentrations of 25, 50, and 100 μg / mL was between 9.3% and 16.3%, which was statistically significant (P<0.01). It can be seen that after the Chinese medicine composition of the present invention intervened in 4T1 cells for 24 hours, a significant difference can be seen from the control group at a low concentration of 25 μg / mL. With the increase of the drug concentration of the Chinese medicine composition of the present invention and the extension of the intervention time, the wound healing rate of 4T1 cells and the healing rate of the control group showed significant dose dependence and statistical difference (P < 0.01 vs Control group; 24h, 48h). It can be seen that the Chinese medicine composition of the present invention has a significant inhibitory effect on both 4T1 and MDA-MB-231 breast cancer cells.

[0069] Table 4. Effect of the Chinese medicine composition of the present invention on the migration ability of MDA-MB-231 cells

[0070]

[0071] ** P<0.01vs Control group (0μg / mL; 24h, 48h). The experimental results of the Chinese medicine composition of the present invention intervening in the wound healing experiment of MDA-MB-231 cells for 24h and 48h.

[0072] Table 5. Effect of the Chinese medicine composition of the present invention on the migration ability of 4T1 cells

[0073]

[0074] ** P<0.01vs Control group (0μg / mL; 24h, 48h). The experimental results of the Chinese medicine composition of the present invention intervening in the 24h and 48h wound healing experiment of 4T1 cells.

[0075] (3) Effect of the Chinese medicine composition of the present invention on the invasion ability of TNBC cells.

[0076] TNBC cells MDA-MB-231 and 4T1 in the logarithmic growth phase were inoculated in 6-well cell culture plates, and blank control groups and groups with different concentrations of the Chinese medicine composition of the present invention (25, 50, 100 μg / mL) were set up for intervention for 24 h and 48 h. Matrigel matrix gel was diluted with serum-free medium at a ratio of 1:8 on ice, and 50 μL per well was added to the upper chamber of the Transwell chamber. After the matrix gel was polymerized into a gel, serum-free medium was added to the upper and lower chambers for overnight hydration. The cell density was adjusted to 5×10 5 cells / mL, 100 μL per well was inoculated in the upper chamber of the Transwell chamber (8 μm pore size), 600 μL of complete medium was added to the lower chamber, and the cells were cultured in a 37°C, 5% CO2 incubator for 12 h. The filter membrane was fixed, stained with crystal violet, photographed under an inverted microscope, and the cells that moved to the lower layer of the membrane were counted. Three fields of view were counted for each sample, and the average value was taken.

[0077] The results of the Transwell invasion experiment showed that as the concentration of the Chinese medicine composition of the present invention increased, the invasion ability of 4T1 and MDA-MB-231 breast cancer cells gradually decreased, showing a more obvious dose-dependency. Compared with the control group 0μg / mL, the group treated with the Chinese medicine composition of the present invention had a significant statistical difference in inhibiting the invasion ability of breast cancer cells (P<0.05, P<0.01vs Control group; 24h, 48h). Figure 5 , as shown in Tables 6-7. For MDA-MB-231 cells, the invasion ability was significantly reduced after intervention with the Chinese medicine composition of the present invention for 24 / 48h. After intervention with the three concentrations of the Chinese medicine composition of the present invention (25μg / mL, 50μg / mL, 100μg / mL) for 24 / 48h, the number of MDA-MB-231 cells that penetrated the chamber was about 33-785 and 136-584; for 4T1 cells, intervention with the Chinese medicine composition of the present invention at low, medium and high concentrations (25μg / mL, 50μg / mL, 100μg / mL) for 24 / 48h could significantly reduce the number of cells that penetrated the chamber. After intervention with different concentrations of Chinese medicine composition for 24 / 48h, the number of cells that penetrated the chamber of 4T1 cells was about 3-679 and 9-307, and the number of cells that penetrated the chamber was negatively correlated with the concentration of the Chinese medicine composition of the present invention. It can be seen that the Chinese medicine composition of the present invention can reduce the invasion ability of TNBC cells.

[0078] Table 6. Effect of the Chinese medicine composition of the present invention on the invasion ability of MDA-MB-231 cells

[0079]

[0080] * P<0.05,** P<0.01vs Control group (0μg / mL; 24h, 48h). The experimental results of the Chinese medicine composition of the present invention intervening in the 24h and 48h Transwell invasion experiment of MDA-MB-231 cells.

[0081] Table 7. Effect of the Chinese medicine composition of the present invention on the invasion ability of 4T1 cells

[0082]

[0083] * P<0.05, ** P<0.01vs Control group (0μg / mL; 24h, 48h). The experimental results of the Chinese medicine composition of the present invention intervening in the 24h and 48h Transwell invasion experiment of 4T1 cells.

[0084] (4) Identification of blood components of the Chinese medicine composition of the present invention.

[0085] The components of the mouse lung tissue of the Chinese medicine composition of the present invention were analyzed by ultra-high performance liquid chromatography Q extraction orbital mass spectrometry (UHPLC-Q Exactive Orbitrap-MS) technology. Methods After the mice were given the Chinese medicine composition of the present invention, blood was collected from the eye sockets and lung tissue was collected. The ACQUITY UPLC HSS T3 column (100mm×2.1mm, 1.8um) was used, and the mobile phase was A-water (containing 0.1% formic acid), B-acetonitrile (flow rate: 0.35mL / min) gradient elution. The mass spectrometer used an electrospray ionization (ESI) ion source to collect sample data in positive and negative ion modes. The components of the Chinese medicine composition of the present invention in the blood and lung tissue of mice were identified by comparing the retention time, accurate mass number, secondary fragments and isotope distribution using Progenesis QI v3.0 software.

[0086] The Chinese medicine composition of the present invention was analyzed by LC-MS. These compounds have been preliminarily identified. Figure 6The UHPLC-MS base peak chromatogram (BPC) spectra of the Chinese medicine composition of the present invention in positive and negative ion modes. We compared the Chinese medicine composition of the present invention with the reference material database (lum / z-TCM) and identified the compounds based on accurate mass-to-charge ratio (m / z), secondary fragments and isotope distribution. Identification is based on three reference dimensions: (1) the error between the retention time and retention time of the standard in the database is within ±0.2min; (2) the primary molecular weight error is within 5ppm; (3) the comparison of the measured MS2 with the standard MS2. Based on the above identification strategy, 913 compounds of the Chinese medicine composition of the present invention were preliminarily identified, including flavonoids, terpenes, organic acids and derivatives, sugars and glycosides, amino acids and peptides, fatty acyl, phenylpropanoids, etc. Among them, 122 components enter the blood and 109 components enter the target. We screened the Chinese medicine components with the top 10 content and level 1 in the Chinese medicine, as well as the identification results of the Chinese medicine components that enter both the blood and the lungs, and displayed them (as shown in Tables 8-10).

[0087] Previous literature reports show that icariin, xanthine, xanthurenic acid, guanosine, and icariin II (Baohuoside I) all have good inhibitory effects on breast tumors, among which icariin II (Baohuoside I) can significantly inhibit breast cancer lung metastasis. Figure 7 , Figure 8 The mass spectrum is shown in the figure, which further confirms the efficacy of the Chinese medicine composition of the present invention in inhibiting breast cancer metastasis.

[0088] Table 8. Identification results of the top 10 components of the Chinese medicine composition of the present invention in terms of blood content

[0089]

[0090] Table 9. Identification results of the top 10 components of the Chinese medicine composition of the present invention that enter the lungs in vivo

[0091]

[0092] Table 10. Identification results of components of the Chinese medicine composition of the present invention that enter both the blood and the lungs in vivo

[0093]

Claims

1. A Chinese medicine composition for treating triple-negative breast cancer lung metastasis, characterized in that: The medicine is prepared from the following raw materials in parts by weight: 3-5 parts of epimedium, 3-5 parts of trillium, 5-15 parts of zedoaria, 2-8 parts of apiary, and 5-15 parts of aconite.

2. The Chinese medicine composition for treating triple-negative breast cancer lung metastasis according to claim 1, characterized in that: The weight portions of the raw materials are respectively: 4 parts of epimedium, 4 parts of trillium, 10 parts of zedoaria, 5 parts of nivalis, and 10 parts of aconite.

3. The Chinese medicine composition for treating lung metastasis of triple-negative breast cancer according to claim 1 or 2, characterized in that: The clinically acceptable oral pharmaceutical preparation is prepared according to conventional Chinese medicine preparation methods.

4. The Chinese medicine composition for treating triple-negative breast cancer lung metastasis according to claim 3, characterized in that: The oral pharmaceutical preparation is a granule.

5. The method for preparing the Chinese medicine composition for treating triple-negative breast cancer lung metastasis according to claim 4, characterized in that: The process steps include: Weigh each raw material drug by weight, add water and boil twice, add 8-12 times the amount of water for the first time, boil for 2-4 hours, add 8-12 times the amount of water for the second time, boil for 1-1.5 hours, combine the decoctions, filter, concentrate the filtrate to a relative density of 1.14-1.15 at 60-80°C, and add pharmaceutically acceptable excipients to prepare granules.

6. Use of the Chinese medicine composition according to any one of claims 1 to 4 in the preparation of a medicament for treating lung metastasis of triple-negative breast cancer.

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