A traditional Chinese medicine composition for treating triple-negative breast cancer lung metastasis, and its preparation method and use

Abstract

The present invention discloses a Chinese medicine composition for treating triple-negative breast cancer lung metastasis, which is prepared from the following raw materials in parts by weight: 3-5 parts of Epimedium, 3-5 parts of Psoralea corylifolia, 5-15 parts of Curcuma zedoaria, 2-8 parts of Vespermum officinale, and 5-15 parts of Rhizoma Aesculapii. The Chinese medicine composition of the present invention has the effects of removing blood stasis and attacking accumulation, soothing the liver and regulating qi, tonifying the kidney and strengthening bones, and dispersing stagnation and detoxifying. It can improve the disease-free survival rate of patients with metastatic breast cancer 2 years after surgery, delay the recurrence and metastasis of breast cancer after surgery, enhance patient immunity, and improve patient clinical symptoms. In addition, the present invention also discloses a preparation method and use of the Chinese medicine composition.

Description

Technical Field

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

[0002] Breast cancer is currently one of the leading causes of cancer-related death in women. Recent studies have found that breast cancer accounts for 11.7% of all tumors and 30% of all cancer cases in women. Triple-negative breast cancers (TNBC) comprise approximately 15%-20% of all invasive breast cancers. Because they lack estrogen receptors (ER), progesterone receptors (PR), and human epidermal growth factor receptor 2 (HER-2), they are insensitive to estrogen receptor antagonists, aromatase inhibitors, trastuzumab, and pertuzumab. Recommended treatment options for TNBC are limited to single-agent sequential chemotherapy or combination chemotherapy. These limitations increase the risk of recurrence, metastasis, and chemotherapy resistance. Over 90% of deaths are due to recurrence and metastasis, with the lungs, bones, liver, and brain being the most common sites of metastasis. Studies have shown that 58.0% of TNBCs present with lung metastases, posing a high risk. Literature reports that the 5-year survival rate for patients with stage III-IV TNBC is only 13%. Currently, while surgery, radiotherapy, chemotherapy, and targeted therapies such as EGFR inhibitors can improve clinical symptoms to some extent, they are also associated with the risk of short-term recurrence, chemotherapy resistance, and severe adverse reactions (gastrointestinal reactions, bone marrow suppression, and hair loss). Traditional Chinese medicine (TCM) is increasingly prominent in the adjuvant treatment of TNBC metastasis. It can complement surgery, radiotherapy, and chemotherapy to modulate immune function, mitigate chemotoxicity, prevent recurrence and metastasis, and prolong survival.

[0003] At present, the main clinical methods of TCM-assisted treatment of TNBC patients' metastasis are dialectical treatment and prescription preparation. There is no effective TCM composition 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 traditional 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 zedoary, 2-8 parts of nivalis, and 5-15 parts of aconite.

[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 Nigella Sativa, and 10 parts of Atractylodes macrocephala.

[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, nectarine, and atractylodes lancea, decocting twice with water, the first time adding 8-12 times the amount of water and decocting for 2-4 hours, the second time adding 8-12 times the amount of water and 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 method further comprises: adding pharmaceutically acceptable excipients to prepare granules.

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

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

[0011] Epimedium: Pungent, sweet, and warm. It enters the liver and kidney meridians. It nourishes the kidneys and strengthens yang, strengthens bones and muscles, and dispels rheumatism, enhancing the endocrine system's secretory function.

[0012] Trillium gracile: Sweet, slightly pungent, and cold. It enters the liver, stomach, and spleen meridians. It has the effects of clearing heat and detoxifying, resolving phlegm and dispersing stagnation, and has multiple pharmacological effects, including anti-tumor, antibacterial, and lipid-lowering.

[0013] Curcuma: Pungent, bitter, and warm. It enters the Liver and Spleen meridians. It promotes blood circulation, promotes Qi flow, eliminates stagnation, and relieves pain. It is commonly used to treat lumps, blood stasis, amenorrhea, and chest pain.

[0014] Beehives: Sweet in flavor, neutral in nature. It enters the liver, stomach, and kidney meridians. It dispels wind and relieves pain, attacks toxins and kills insects, relieves itching, and eliminates dampness and toxins. It is commonly used to treat sores, swellings, mastitis, scrofula, and stubborn skin rashes.

[0015] August Zha: Bitter in flavor, cold in nature. It enters the Liver and Stomach meridians. It soothes the liver and regulates Qi, promotes blood circulation and relieves pain, and promotes urination. It also enhances immunity, has anti-inflammatory properties, and inhibits bacteria.

[0016] The traditional Chinese medicine composition for treating TNBC lung metastasis of the present invention uses Trillium gracile and Beehive as the main herbs, both of which have strong anti-cancer and stagnation-dispersing effects and have the effect of "fighting poison with poison". At the same time, Beehive also has the effect of warming the kidney; Curcuma zedoaria is used as the auxiliary herb to break up blood stasis and attack accumulation, thereby achieving the effect of eliminating tumors and dispersing stagnation; Epimedium and Atractylodes macrocephala are used as adjuvants to soothe the liver and regulate qi, nourish the kidney and strengthen the bones, and enable the whole formula to dispel evil without harming the body, thereby better exerting the function of dispersing stagnation and detoxification.

[0017] Subsequent test cases will demonstrate that the Chinese herbal 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 both 4T1 and MDA-MB-231 breast cancer cells; can reduce the invasive ability of TNBC cells; and has a significant therapeutic effect in inhibiting breast cancer lung metastasis. BRIEF DESCRIPTION OF THE DRAWINGS

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

[0019] Figure 2 The effect of the Chinese medicine composition of the present invention on lung metastases in mice with breast cancer lung metastasis 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. ** 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 < 0.01 vs Control group (0 μg / mL; 24 h, 48 h); A, C: Experimental results and statistical graphs of the wound healing experiment on MDA-MB-231 cells treated with the Chinese herbal composition of the present invention for 24 h and 48 h; B, D: Experimental results and statistical graphs of the wound healing experiment on 4T1 cells treated with the Chinese herbal composition of the present invention for 24 h and 48 h.

[0022] Figure 5 The effect of the Chinese medicine composition of the present invention on the invasion ability of TNBC cells [* 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 traditional 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 traditional Chinese medicine composition of the present invention].

[0023] Figure 6 The base peak chromatogram (BPC) of the traditional 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] is shown.

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

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

[0026] The present invention will be further described below in conjunction with the accompanying drawings and specific examples. These embodiments should be understood to be merely illustrative of the present invention and not intended to limit the scope of protection of the present invention. After reading the contents described herein, those skilled in the art may make various changes or modifications to the present invention, and these equivalent variations 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 macrocephala, 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 macrocephala, 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 macrocephala, 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 macrocephala, 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 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.

[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 macrocephala, 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 Trial

[0040] 1. Experimental Drugs

[0041] During the preparation process of the Chinese medicine composition described in Example 6, the decoctions were 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 occurred 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 included: ① Primary breast cancer that underwent surgical treatment and was confirmed as a malignant epithelial breast tumor (invasive breast cancer) by pathological examination; ② The primary surgery occurred between May 1, 2019, and May 31, 2022; ③ Patients with clinical stage I-III operable disease and metastasis detected by postoperative pathology; ④ Female patients aged 18 to 90 years; ⑤ Patients who voluntarily participated in this trial and took the Chinese medicine composition of the present invention for ≥6 months. The 2-year disease-free survival (DFS) of the patients was observed: the ratio of patients who had no disease-related recurrence or metastasis within 2 years from the time of surgical treatment was calculated.

[0044] The study results showed that the two-year DFS rate for the treatment group reached 89.80% (Table 1), and the two-year DFS rate for patients with isolated lymph node metastasis reached 95.23%. Existing literature reports that the two-year DFS rate for patients with non-metastatic breast cancer is 83.50%, which is significantly lower than the efficacy of patients with metastatic breast cancer in this study. This finding suggests that the traditional 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] Experimental Example 2 - In vivo efficacy observation [Effect of the Chinese medicine composition of the present invention on tumor volume and weight in 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 Chinese medicine composition of the present invention (0.85g / kg), medium-dose group of Chinese medicine composition prepared in Example 6 of the present invention (1.7g / kg), high-dose group of Chinese medicine composition prepared in Example 6 of the present invention (2.55g / kg), paclitaxel-positive drug group (0.01g / kg). Each group started administration 14 days after inoculation of 4T1 cells. The low, medium and high-dose groups of Chinese medicine composition of the present invention were gavage-fed for 21 consecutive days, once a day; paclitaxel-positive drug group 0.01g / kg, intraperitoneal injection, once a week. The normal group and the control group were gavage-fed 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 tumors in mice (P < 0.01); the lung tissue of mice soaked in picric acid ( Figure 2 ), it was found that compared with the model group, the group treated with the Chinese medicine composition of the present invention was able to significantly inhibit lung metastasis in 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 drug 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 Macrocephala 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 then mixing all the medicinal liquids, filtering, evaporating, and drying.

[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 into 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. After 24, 48, and 72 hours of intervention, the CCK8 method was used to detect 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-dependent effect, indicating that the drug can inhibit the proliferation of two TNBC cells (such as Figure 3 , as shown in Table 2-3).

[0060] Table 2. Effects 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. Effects 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 MDA-MB-231 cells and mouse breast cancer 4T1 cells in the logarithmic growth phase were inoculated into 6-well cell culture plates and subjected to cell scratching. Basal culture medium or 25, 50, and 100 μg / mL of the Chinese medicinal composition of the present invention were added, respectively, and cultured for 24 h and 48 h. Pictures were taken under an inverted microscope, and the healing area and migration rate of the scratched 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 24 and 48 hours of intervention (P<0.01, Figure 4, Tables 4-5). For MDA-MB-231 cells, the cell migration rate was 1.8%-13.6% after 24 hours of treatment with 100 μg / mL of the Chinese herbal composition of the present invention, and 8.8%-12.8% after 48 hours of treatment, both showing significant statistical differences compared to the control group (P < 0.01). For 4T1 cells, after 24 hours of treatment with the Chinese herbal composition of the present invention, the cell migration rate at concentrations of 25, 50, and 100 μg / mL was only 5.6%-10.5%, showing significant statistical differences compared to the control group (P < 0.01). After 48 hours of treatment, the cell migration rates at concentrations of 25, 50, and 100 μg / mL were between 9.3% and 16.3%, showing significant statistical differences compared to the blank control group (P < 0.01). As can be seen, after 24 hours of treatment with the Chinese herbal composition of the present invention, a significant difference was observed compared to the control group at a low concentration of 25 μg / mL. With increasing concentrations of the Chinese herbal composition of the present invention and prolonged treatment time, the wound healing rate of 4T1 cells showed a significant dose-dependent and statistically significant difference compared to the healing rate of the control group (P < 0.01 vs Control group; 24 hours, 48 ​​hours). This indicates that the Chinese herbal composition of the present invention has a significant inhibitory effect on both 4T1 and MDA-MB-231 breast cancer cells.

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

[0070]

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

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

[0073]

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

[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 into 6-well cell culture plates, and blank control groups and groups treated with different concentrations of the Chinese medicine composition of the present invention (25, 50, and 100 μg / mL) were set up for 24 hours and 48 hours. Matrigel 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 matrigel 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 into the upper chamber of a Transwell chamber (8 μm pore size), and 600 μL of complete medium was added to the lower chamber. The cells were cultured in a 37°C, 5% CO2 incubator for 12 h. The filters were fixed and stained with crystal violet. The cells were photographed under an inverted microscope, and the cells that migrated to the lower layer of the membrane were counted. Three fields of view were counted for each sample, and the average value was calculated.

[0077] The results of the Transwell invasion assay showed that as the concentration of the Chinese herbal composition of the present invention increased, the invasive ability of both 4T1 and MDA-MB-231 breast cancer cells gradually decreased, showing a significant dose-dependency. Compared with the control group (0 μg / mL), the group treated with the Chinese herbal composition of the present invention had a statistically significant difference in inhibiting the invasive ability of breast cancer cells (P < 0.05, P < 0.01 vs Control group; 24h, 48h). Figure 5 As shown in Tables 6-7. For MDA-MB-231 cells, intervention with the Chinese herbal composition of the present invention for 24 / 48 hours also significantly reduced their invasive ability. Without intervention with the three concentrations of the Chinese herbal composition of the present invention (25 μg / mL, 50 μg / mL, 100 μg / mL) for 24 / 48 hours, the number of MDA-MB-231 cells that permeated the chamber was approximately 33–785 and 136–584 respectively. For 4T1 cells, intervention with the low, medium, and high concentrations of the Chinese herbal composition of the present invention (25 μg / mL, 50 μg / mL, 100 μg / mL) for 24 / 48 hours significantly reduced the number of cells that permeated the chamber. After intervention with different concentrations of the Chinese herbal composition for 24 / 48 hours, the number of 4T1 cells that permeated the chamber was approximately 3–679 and 9–307 respectively. The number of cells that permeated the chamber was negatively correlated with the concentration of the Chinese herbal composition of the present invention. Thus, the Chinese herbal composition of the present invention can reduce the invasive ability of TNBC cells.

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

[0079]

[0080] * P<0.05,** P<0.01 vs Control group (0 μg / mL; 24h, 48h). Experimental results of the intervention of the Chinese herbal composition of the present invention on the Transwell invasion assay of MDA-MB-231 cells for 24h and 48h.

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

[0082]

[0083] * P<0.05, ** P<0.01 vs Control group (0 μg / mL; 24h, 48h). Experimental results of the 24h and 48h Transwell invasion assay of 4T1 cells treated with the Chinese herbal composition of the present invention.

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

[0085] The components of the mouse lung tissue containing the traditional Chinese medicine composition of the present invention were analyzed using ultra-high performance liquid chromatography-Q extraction orbitrap mass spectrometry (UHPLC-Q Exactive Orbitrap-MS). Methods: After administration of the traditional Chinese medicine composition of the present invention to mice, blood was collected from the eye sockets and lung tissue was collected. The analysis was performed on an ACQUITY UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 μm) using a mobile phase consisting of A: water (containing 0.1% formic acid) and B: acetonitrile (flow rate: 0.35 mL / min) in a gradient elution. The mass spectrometer used an electrospray ionization (ESI) source, acquiring sample data in positive and negative ion modes. The components of the traditional Chinese medicine composition of the present invention in the mouse blood and lung tissue were identified by comparing retention times, accurate masses, secondary fragments, and isotopic 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 following are the base peak chromatograms (BPC) of the UHPLC-MS of the Chinese herbal composition of the present invention in positive and negative ion modes. We compared the Chinese herbal 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 was based on three reference dimensions: (1) the error between the retention time and retention time of the standard in the database was within ±0.2min; (2) the error of the primary molecular weight was within 5ppm; (3) the comparison of the measured MS2 with the standard MS2. Based on the above identification strategy, 913 compounds of the Chinese herbal composition of the present invention were preliminarily identified, including flavonoids, terpenes, organic acids and derivatives, sugars and glycosides, amino acids and peptides, fatty acyl compounds, phenylpropanoids, etc. Among them, 122 components entered the blood and 109 components entered the target. We screened the Chinese herbal components that were in the top 10 in terms of content and level 1 in the Chinese herbal medicine, as well as the Chinese herbal components that entered both the blood and the lungs, and displayed the identification results (as shown in Tables 8-10).

[0087] Previous literature reports have shown 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 FIG. This 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 that have the highest 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

[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

[0093]

Claims

1. A Chinese medicine composition for treating triple-negative breast cancer lung metastasis, characterized in that: The drug 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 zedoary, 2-8 parts of beehive, 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 zedoary, 5 parts of nivalis, and 10 parts of aconite.

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

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 by weight, add water and boil twice, add 8-12 times the amount of water for the first time and boil for 2-4 hours, add 8-12 times the amount of water for the second time and boil for 1-1.5 hours, combine the decoctions, filter, and 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 traditional 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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