Traditional Chinese medicine composition for treating triple negative breast cancer and application thereof

Abstract

The invention belongs to the field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine composition for treating triple negative breast cancer and application thereof. The traditional Chinese medicine composition is prepared from the following main active ingredients in parts by mass: 20 to 40 parts of radix astragali seu hedysari, 10 to 20 parts of rhizoma curcumae, 10 to 20 parts of herba hedyotidis diffusae, 5 to 15 parts of fructus aurantii immaturus, 5 to 15 parts of radix bupleuri, 5 to 15 parts of radix paeoniae alba and 5 to 15 parts of liquorice root. Animal experiments show that the traditional Chinese medicine composition can significantly inhibit tumor growth and pulmonary metastasis of triple-negative breast cancer tumor-bearing mice, increase the proportion of CD8 + T lymphocytes in blood, reduce the proportion of MDSCs, significantly down-regulate the expression of pro-inflammatory cytokines such as IL-6, IL-1alpha, CXCL1 and GM-CSF in serum, and cooperate with PD-L1 immunotherapy to achieve a synergistic effect. And the whole blood and liver and kidney function indexes are in a normal range. Therefore, the traditional Chinese medicine composition disclosed by the invention can be used for effectively inhibiting the tumor growth and pulmonary metastasis of the triple negative breast cancer, remodeling the immune state of an organism and reducing the release of proinflammatory factors, and is safe, reliable, free of obvious liver and kidney toxicity and blood toxicity and small in side effect.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine, specifically relating to a traditional Chinese medicine composition for treating triple-negative breast cancer and its application. Background Technology

[0002] Triple-negative breast cancer (TNC) is a subtype of breast cancer that expresses negative values ​​for estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2), accounting for 10% to 20% of all breast cancers. TNC is the most malignant subtype with the worst prognosis, characterized by: early onset (usually under 40 years old), rapid tumor growth, high invasiveness, high malignancy, a high tendency for distant metastasis, a high recurrence rate, low sensitivity to conventional treatments, and a tendency to develop drug resistance, leading to extremely poor prognosis. The 5-year survival rate for TNC is 12% to 20% lower than other subtypes; once metastasized, the 5-year survival rate for metastatic TNC is only about 22%.

[0003] Triple-negative breast cancer (TNC) is a challenging area in breast cancer treatment and a clinical bottleneck limiting overall survival. Because it is negative for ER, PR, and HER2 expression, endocrine therapy targeting these markers and HER2-targeted therapy are ineffective. Currently, chemotherapy is the first-line treatment for TNC, but it has significant clinical limitations, with a clinicopathological complete response rate of only about one-third, and suffers from severe side effects and poor treatment adherence. First-line treatment regimens, such as combination chemotherapy with taxanes and anthracyclines, have limited efficacy and severe side effects, increasing the risk of cardiotoxicity, secondary leukemia, and myelodysplastic syndromes. The use of carboplatin increases the incidence of neutropenia and thrombocytopenia; bevacizumab may cause serious adverse reactions such as hypertension, infection, thromboembolism, and bleeding. While new drugs such as PARP inhibitors and immune checkpoint inhibitors have emerged in recent years, their application is limited to patients with specific gene mutations (such as BRCA) or PD-L1 positive expression, resulting in a limited beneficiary population. They also face problems such as drug resistance and specific immune-related adverse events, with overall efficacy remaining low and costs high. Therefore, current treatments for triple-negative breast cancer fall far short of clinical needs, and there is an urgent need to develop safer and more effective new anti-triple-negative breast cancer treatment strategies to improve the prognosis of triple-negative breast cancer treatment.

[0004] Traditional Chinese medicine (TCM) has a long history and unique theoretical system in the treatment of malignant tumors, emphasizing the holistic view of "strengthening the body's resistance and eliminating pathogenic factors" and "treatment based on syndrome differentiation." Among these, TCM compound formulas, through the synergistic effects of multiple components, targets, and pathways, can inhibit tumor growth while regulating the body's immune function, improving physical condition, and reducing the toxic side effects of Western medicine treatments—a process known as "reducing toxicity and increasing efficacy." This demonstrates unique advantages in improving patients' quality of life and prolonging survival. Unlike the approach of extracting effective components from single herbs (such as cryptotanshinone in Danshen and saikosaponins in Bupleurum), TCM compound formulas better align with the essence of TCM theory. The precise combination of each component, with its principal, assistant, and adjuvant herbs, produces an overall therapeutic effect greater than the simple sum of individual components. TCM compound formulas offer a new approach to addressing the clinical treatment bottlenecks of triple-negative breast cancer, possessing significant clinical application value and broad development prospects. Currently, there is still an urgent clinical need for a TCM composition that can effectively treat triple-negative breast cancer. Summary of the Invention

[0005] The first objective of this invention is to provide a traditional Chinese medicine composition for treating triple-negative breast cancer, and the second objective of this invention is to provide the application of this traditional Chinese medicine composition.

[0006] According to a first aspect of the present invention, a traditional Chinese medicine composition for treating triple-negative breast cancer is provided, the main active ingredients comprising, by weight: 20-40 parts of Astragalus membranaceus, 10-20 parts of Curcuma zedoaria, 10-20 parts of Hedyotis diffusa, 5-15 parts of Citrus aurantium, 5-15 parts of Bupleurum chinense, 5-15 parts of Paeonia lactiflora, and 5-15 parts of Glycyrrhiza uralensis.

[0007] This invention posits that triple-negative breast cancer is based on "deficiency of vital energy" as its pathogenesis, with "phlegm, blood stasis, and toxins" as key pathological products and pathogenic factors, forming a complex pathogenesis of deficiency in the root and excess in the branch, and a mixture of deficiency and excess. The treatment principle is "strengthening the body's resistance and eliminating pathogenic factors," and the method is "tonifying qi and blood, clearing cancer toxins, and regulating the liver and spleen."

[0008] The herbal composition of this invention uses Astragalus membranaceus and Curcuma zedoaria as the principal herbs. Astragalus membranaceus invigorates qi and blood, strengthens the body's resistance, and fights cancer, while Curcuma zedoaria promotes qi circulation, breaks up blood stasis, eliminates stagnation, and relieves pain. Together, they invigorate the spleen, invigorate qi, break up blood stasis, and inhibit tumor recurrence in triple-negative breast cancer. Hedyotis diffusa and Citrus aurantium are used as assistant herbs, which work synergistically with the principal herbs to detoxify, fight cancer, resolve phlegm, dissipate stagnation, and promote the flow of qi and blood. Bupleurum chinense and Paeonia lactiflora are used as adjuvant herbs to soothe the liver, regulate qi, and nourish blood, working together with the principal and assistant herbs to replenish qi and blood. Glycyrrhiza uralensis is used as the guiding herb to invigorate the spleen, clear heat, detoxify, harmonize the herbs, and mitigate the strong effects of Curcuma zedoaria and other herbs. The seven herbs combined work synergistically to detoxify, fight cancer, invigorate qi and blood, soothe the liver, and invigorate the spleen, effectively treating triple-negative breast cancer. The raw materials for this formula come from the following sources: Astragalus: The root of Astragalus memeranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao or Astragalus membranaceus (Fisch.) Bge., belonging to the genus Astragalus of the legume family.

[0009] Curcuma: The rhizome of Curcuma aeruginosa Roxb. [C. zedoaria nonRosc.], Curcuma kwang siensis SGLee et CFLiang, and Curcuma wenyujin YHChen et C.Ling, all belonging to the genus Curcuma of the ginger family (Zingiberaceae).

[0010] Hedyotis diffusa Willd. (family Rubiaceae): The whole herb of Hedyotis diffusa Willd.

[0011] Citrus aurantium L. (citrus aurantium L.) and its cultivated varieties or Citrus sinensis Osbeck (citrus sinensis Osbeck) are dried young fruits of plants in the Rutaceae family.

[0012] Bupleurum: The dried root of Bupleurum chinense DC or Bupleurum corzonerifolium Willd, both belonging to the Apiaceae family.

[0013] White peony root: The root of Paeonia lactiflora Pall., a plant belonging to the genus Paeonia in the family Ranunculaceae.

[0014] Licorice: The dried root and rhizome of Glycyrrhiza uralensis Fisch., Glycyrrhiza inflata Bat., or Glycyrrhiza glabra L., plants belonging to the genus Glycyrrhiza in the legume family.

[0015] In some embodiments, the main active ingredients, by weight, include: 30 parts Astragalus membranaceus, 15 parts Curcuma zedoaria, 15 parts Hedyotis diffusa, 10 parts Citrus aurantium, 10 parts Bupleurum chinense, 10 parts Paeonia lactiflora, and 10 parts Glycyrrhiza uralensis.

[0016] In some implementations, the licorice is prepared as roasted licorice.

[0017] In some implementations, pharmaceutically acceptable excipients are also included.

[0018] In some embodiments, the excipient is at least one of fillers, suspending agents, disintegrants, lubricants, binders, preservatives, sweeteners, and flavoring agents, to help improve the stability or activity of the drug, or to produce an acceptable taste or odor when taken orally.

[0019] In some embodiments, the dosage form of the traditional Chinese medicine composition is a decoction, oral solution, mixture, paste, granules, lyophilized powder, powder, tablet, pill, capsule, sustained-release preparation, controlled-release preparation, ointment, or suppository. This facilitates clinical administration.

[0020] According to a second aspect of the present invention, the use of the above-described traditional Chinese medicine composition for treating triple-negative breast cancer in the preparation of a medicament for treating triple-negative breast cancer is provided.

[0021] According to a third aspect of the present invention, the above-described traditional Chinese medicine composition for treating triple-negative breast cancer is provided for use in the preparation of a medicament that enhances the efficacy of PD-L1 monoclonal antibodies in inhibiting tumor growth and lung metastasis. Specifically, the tumor is a triple-negative breast cancer tumor.

[0022] The beneficial effects of this invention include: (1) Animal experiments showed that the traditional Chinese medicine composition of the present invention can significantly inhibit tumor growth and lung metastasis in triple-negative breast cancer-bearing mice, and significantly increase CD8 levels in the blood of triple-negative breast cancer-bearing mice. + The proportion of T lymphocytes was reduced, the proportion of MDSCs was decreased, and the expression of pro-inflammatory cytokines such as IL-6, IL-1α, CXCL1, and GM-CSF in the serum of triple-negative breast cancer-bearing mice was significantly downregulated, while whole blood and liver and kidney function indicators were within the normal range. This suggests that the traditional Chinese medicine composition of the present invention can effectively inhibit the growth and lung metastasis of triple-negative breast cancer, reshape the body's immune status, reduce the release of pro-inflammatory factors, and is safe and reliable with no significant liver, kidney, or blood toxicity and few side effects.

[0023] (2) Animal experiments showed that the combination of the traditional Chinese medicine composition of the present invention and PD-L1 monoclonal antibody can synergistically inhibit tumor growth and lung metastasis in triple-negative breast cancer-bearing mice. This suggests that the traditional Chinese medicine composition of the present invention can synergistically enhance the efficacy of immunotherapy with PD-L1 monoclonal antibody, regulate the tumor immune microenvironment and the level of inflammatory factors in the body, and synergistically inhibit the growth of triple-negative breast cancer tumors and lung metastasis.

[0024] (3) The traditional Chinese medicine composition of the present invention has been used clinically for many years with good clinical efficacy. It can not only significantly relieve fatigue symptoms in patients with triple-negative breast cancer and improve their quality of life, but also play a role in preventing recurrence and metastasis of triple-negative breast cancer. The traditional Chinese medicine composition of the present invention can be applied to the entire course of treatment of triple-negative breast cancer, including the perioperative period, perichemotherapy period, periradiotherapy period, targeted therapy period, and consolidation therapy period. Attached Figure Description

[0025] Figure 1 This is a diagram showing the results of tumor growth in triple-negative breast cancer-bearing mice in animal experiment one of this invention, wherein... Figure 1 A shows photographs of mouse tumors, tumor growth curves, and tumor weight comparison charts. Figure 1 B represents the results of Ki-67 immunohistochemistry and TUNEL cell apoptosis experiments in mouse tumor tissue. * indicates P < 0.05 compared to the model group.

[0026] Figure 2 These are photographs of mouse lungs from animal experiment one of this invention, and the results of HE staining of lung tissue pathological sections.

[0027] Figure 3 The CD8+ levels in the peripheral blood of mice in each group of animal experiments of this invention are... + Flow cytometry analysis results of T lymphocytes and myeloid-derived immunosuppressive cells, among which, Figure 3 A represents the CD8 concentration in the peripheral blood of mice in each group. + The percentage of T lymphocytes; Figure 3 B represents the percentage of myeloid-derived immunosuppressive cells in the peripheral blood of mice in each group. ** indicates P < 0.01 compared to the model group.

[0028] Figure 4 This is a graph showing the results of pro-inflammatory cytokine expression in the serum of triple-negative breast cancer-bearing mice in animal experiment one of this invention. Figure 4 A represents the expression result of IL-6; Figure 4 B represents the expression results of IL-1α, CXCL1, and GM-CSF. * indicates P < 0.05 compared to the model group, and ** indicates P < 0.01 compared to the model group.

[0029] Figure 5 These are the results of whole blood analysis and liver and kidney function index detection in animal experiment one of this invention, wherein... Figure 5 A represents the results of WBC, Hb, and PLT tests. Figure 5 B represents the detection results of ALT, AST, Urea, and Cr. * indicates P < 0.05 compared to the model group, and ** indicates P < 0.01 compared to the model group.

[0030] Figure 6 These are the test results from animal experiment two of the present invention, wherein, Figure 6 A is a graph showing the growth curve of mouse tumors and a comparison of tumor weight; Figure 6 B represents the immunohistochemical detection result of Ki-67 index in mouse tumor tissue; Figure 6 C is a photograph of a mouse's lung; Figure 6 D represents the HE staining results of mouse lung tissue. * indicates P < 0.05 compared to the model group. Detailed Implementation

[0031] The present invention will be further described in detail below with reference to embodiments. It is worth noting that the following embodiments are only for better explaining the content of the present invention and do not limit the scope of protection of the present invention. Process steps not disclosed in the embodiments are prior art. Unless otherwise specified, all raw materials and reagents used in the present invention are commercially available.

[0032] Example 1 The traditional Chinese medicine decoction for treating triple-negative breast cancer in this embodiment is prepared from the following raw materials: Astragalus membranaceus 30g, Curcuma zedoaria 15g, Hedyotis diffusa 15g, Citrus aurantium 10g, Bupleurum chinense 10g, Paeonia lactiflora 10g, and Glycyrrhiza uralensis 10g.

[0033] The preparation method includes the following steps: mix the above-mentioned Chinese medicinal materials according to the ratio, add about 1500 mL of water, soak for half an hour, then decoct for 1 hour, filter, add water to the dregs and decoct twice more, mix the decoction filtered from the three decoctions, and concentrate the decoction to 1000 mL.

[0034] Dosage and administration: Drink 1000mL (equivalent to 100g of raw medicine) daily, divided into three doses.

[0035] Example 2 The traditional Chinese medicine granules for treating triple-negative breast cancer in this embodiment are prepared from the following raw materials: Astragalus membranaceus 30g, Curcuma zedoaria 15g, Hedyotis diffusa 15g, Citrus aurantium 10g, Bupleurum chinense 10g, Paeonia lactiflora 10g, and Glycyrrhiza uralensis 10g.

[0036] The preparation method includes the following steps: the above-mentioned Chinese medicinal materials are pulverized, passed through a No. 1 sieve, and extracted three times by reflux with 12 times, 10 times and 8 times the amount of 95% ethanol, respectively, for 1 hour each time. After filtration, the filtrate is allowed to stand, and the supernatant is taken. The supernatants obtained from the three extractions are combined, concentrated under reduced pressure to form a paste, and the ethanol is evaporated to obtain the raw material extract. Then, excipients such as property agents and flavoring agents are added to make granules, which are then packaged according to the single dose to obtain granules.

[0037] Example 3 The traditional Chinese medicine granules for treating triple-negative breast cancer in this embodiment are prepared from the following raw materials: Astragalus membranaceus 30g, Curcuma zedoaria 15g, Hedyotis diffusa 15g, Citrus aurantium 10g, Bupleurum chinense 10g, Paeonia lactiflora 10g, and Glycyrrhiza uralensis 10g.

[0038] The preparation method includes the following steps: the above-mentioned Chinese medicinal materials are put into an extraction tank, and decocted for 2 hours with 8 times the amount of water and 1 hour with 7 times the amount of water. The extracts from the two extractions are combined, filtered, and the filtrate is concentrated under reduced pressure to a paste. An appropriate amount of maltodextrin is added and stirred evenly, and then spray-dried to obtain spray-dried powder. The spray-dried powder is sieved, and an appropriate amount of maltodextrin is added and mixed evenly. Finally, it is granulated by a dry granulation machine and packaged according to the single dose to obtain granules.

[0039] Example 4 The compound for treating triple-negative breast cancer in this embodiment is prepared from the following raw materials: Astragalus membranaceus 30g, Curcuma zedoaria 15g, Hedyotis diffusa 15g, Citrus aurantium 10g, Bupleurum chinense 10g, Paeonia lactiflora 10g, Glycyrrhiza uralensis 10g.

[0040] Its preparation method includes the following steps: The above-mentioned Chinese medicinal materials are pulverized, passed through a No. 1 sieve, and extracted with 12 times, 10 times, and 8 times the amount of distilled water, respectively, for 1 hour each time. After filtration, the filtrates obtained from the three extractions are combined, and the filtrates are concentrated under reduced pressure. Then, flavoring agents, antibacterial agents, and other excipients are added, and the mixture is packaged into containers of appropriate specifications according to the dosage to obtain the compound preparation.

[0041] Example 5 The freeze-dried powder for treating triple-negative breast cancer in this embodiment is prepared from the following raw materials: Astragalus membranaceus 30g, Curcuma zedoaria 15g, Hedyotis diffusa 15g, Citrus aurantium 10g, Bupleurum chinense 10g, Paeonia lactiflora 10g, and Glycyrrhiza uralensis 10g.

[0042] Its preparation method includes the following steps: The above-mentioned Chinese medicinal materials were put into a reflux extraction tank and extracted three times with 1000mL of water for 1 hour each time. The extracts were filtered through gauze and the filtrates from the three extractions were combined. The supernatant was collected after centrifugation and then concentrated by rotary evaporation to obtain an extract. The extract was then frozen overnight at -80℃ and freeze-dried using a freeze dryer to obtain freeze-dried powder.

[0043] Example 6 The traditional Chinese medicine decoction for treating triple-negative breast cancer in this embodiment is prepared from the following raw materials: Astragalus membranaceus 40g, Curcuma zedoaria 10g, Hedyotis diffusa 20g, Citrus aurantium 5g, Bupleurum chinense 5g, Paeonia lactiflora 15g, and Glycyrrhiza uralensis 15g.

[0044] Its preparation method is the same as that of Example 1.

[0045] Example 7 The traditional Chinese medicine decoction for treating triple-negative breast cancer in this embodiment is prepared from the following raw materials: Astragalus membranaceus 20g, Curcuma zedoaria 20g, Hedyotis diffusa 10g, Citrus aurantium 15g, Bupleurum chinense 15g, Paeonia lactiflora 5g, and Glycyrrhiza uralensis 5g.

[0046] Its preparation method is the same as that of Example 1.

[0047] Animal Experiment 1 A triple-negative breast cancer mouse model was established using BALB / c mice to investigate the safety and efficacy of the traditional Chinese medicine composition of the present invention in treating triple-negative breast cancer.

[0048] I. Grouping, Modeling, and Drug Administration Female BALB / c mice aged 6–8 weeks, weighing 15–18 g, were housed in an SPF environment with a temperature of 22–26°C and a relative humidity of 40%–60%, and were provided with sterilized food and water. After the animals acclimatized to the environment, they were randomly divided into a normal control group (Control), a model group (Model), a low-dose administration group (ADQ807-low), and a high-dose administration group (ADQ807-high), with 6 mice in each group.

[0049] Mice in the model group, low-dose group, and high-dose group were simultaneously inoculated with tumor masses derived from the triple-negative breast cancer cell line 4T1 on the fourth pair of mammary fat pads on the left side of the mice.

[0050] When the tumor diameter reached approximately 0.5 cm, the drugs were administered to the mice in each group according to the following protocol: Low-dose administration group: The lyophilized powder aqueous solution (concentration of 1 g / mL) of Example 5 was administered by gavage at a dose of 3.7 g / kg / d and a gavage volume of 74 μL. High-dose administration group: The lyophilized powder aqueous solution (concentration of 2 g / mL) of Example 5 was administered by gavage at a dose of 7.4 g / kg / d and a gavage volume of 74 μL. Mice in the normal control group and the model group were given 74 μL of physiological saline by gavage.

[0051] Mice in each group were administered the drug once daily by gavage for 30 consecutive days.

[0052] The long and short axes of mouse tumors were measured every 3 days during the drug administration period. At the end of the experiment, mice were sacrificed in accordance with animal ethics requirements, and tumors and lungs were collected. After weighing the tumors, the tumor and lung tissues were preserved for subsequent studies.

[0053] II. Statistical Methods This study used SPSS 26.0 for statistical data analysis. Qualitative data were described by frequency and proportion; quantitative data were described by number of cases, mean ± standard deviation (if normally distributed), or median (if non-normally distributed). If the variation follows (or approximately follows) a normal distribution, a paired-samples t-test was used; if the variation does not follow a normal distribution, the Wilcoxon Signed-Rank Test was used as a robust non-parametric test. A p-value < 0.05 was considered statistically significant.

[0054] III. Test Results 1. Effect of inhibiting tumor growth in triple-negative breast cancer-bearing mice Observe the size of mouse tumors and plot tumor growth curves. After 30 days of drug administration, it was observed that the tumor volume and weight in the high-dose group were significantly lower than those in the model group. Figure 1 A). Further Ki-67 immunohistochemistry and TUNEL cell apoptosis assays were performed on mouse tumor tissues. The results showed that the number of Ki-67 positive cells was lower in the low-dose and high-dose drug groups than in the model group, and the number of apoptotic cells stained by TUNEL assay was higher in the high-dose drug groups than in the model group. Figure 1 B).

[0055] The above results indicate that the traditional Chinese medicine composition of the present invention can significantly inhibit the growth of triple-negative breast cancer xenografts, and the mechanism may be related to its ability to reduce the proliferative activity of triple-negative breast cancer cells and induce their apoptosis.

[0056] 2. Effect of inhibiting lung metastasis in triple-negative breast cancer-bearing mice Lung tissue was collected from mice, and observation revealed that the number of lung metastatic lesions in the low-dose and high-dose drug groups was less than that in the model group. After HE staining of mouse lung tissue sections, the area of ​​lung metastatic lesions in the low-dose and high-dose drug groups was also observed to be smaller than that in the model group under a microscope. Figure 2 ).

[0057] The above results indicate that the traditional Chinese medicine composition of the present invention can inhibit lung metastasis of triple-negative breast cancer in mice.

[0058] 3. The situation of immune regulation The immune system plays an important regulatory role in the occurrence and development of tumors. In this experiment, flow cytometry was used to detect CD8 levels in the peripheral blood of mice in each group. + The proportions of T lymphocytes and myeloid-derived immunosuppressive cells (MDSCs). Flow cytometry analysis showed that the model group had CD8... +The proportion of T lymphocytes was significantly lower than that of the control group (P < 0.01), while the proportion of MDSCs was significantly higher than that of the control group (P < 0.01). Peripheral blood CD8+ cells were significantly higher in both the high-dose and low-dose groups. + The proportion of T lymphocytes increased significantly compared with the model group, and this trend became more pronounced with increasing dosage of the traditional Chinese medicine composition. Figure 3 A). On the other hand, the proportion of peripheral blood MDSCs in the high-dose and low-dose groups was significantly reduced compared with the model group, and the trend of change became more pronounced with the increase of the dosage of the traditional Chinese medicine composition. Figure 3 B).

[0059] The above results indicate that the traditional Chinese medicine composition of the present invention can significantly increase CD8 levels in the blood of triple-negative breast cancer-bearing mice. + The reduced proportion of T lymphocytes and MDSCs suggests that the herbal composition of this invention can inhibit the growth and metastasis of triple-negative breast cancer through immunomodulatory effects.

[0060] 4. Downregulation of pro-inflammatory cytokine expression in the serum of triple-negative breast cancer-bearing mice Inflammatory factors are closely related to the occurrence and development of tumors. Some pro-inflammatory cytokines can accelerate the growth and metastasis of triple-negative breast cancer and inhibit anti-tumor immune responses through various mechanisms. In this experiment, the Quantibody MouseCytokine Array 1 kit (RayBiotech, Inc., Norcross, GA, USA) was used to detect the concentrations of 20 cytokines in the serum of mice in the model group and the high-dose administration group. These cytokines included granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon-γ (IFN-γ), IL-1α, IL-1β, IL-6, chemokine (CXC motif) ligand 1 (CXCL1), chemokine ligand 2 (CCL2), macrophage colony-stimulating factor (M-CSF), tumor necrosis factor-α (TNF-α), and vascular endothelial growth factor (VEGF-A). The RayBiotech data analysis software corresponding to this kit was used for data analysis. The results showed that, compared with the model group, the expression of interleukin-6 (IL-6) in the high-dose treatment group was significantly downregulated, and the difference was statistically significant (P < 0.01). Figure 4 A). Furthermore, compared to the model group, the expression of IL-1α, CXCL1, and GM-CSF in the high-dose treatment group was also reduced to varying degrees, with significant differences (P < 0.05). Figure 4 B).

[0061] The above results suggest that the inhibitory effect of the traditional Chinese medicine composition of the present invention on tumor growth and metastasis in triple-negative breast cancer-bearing mice can be attributed to its regulation of the body's immune function and reduction of inflammatory activity.

[0062] 5. Safety Complete blood analysis and liver and kidney function tests were performed on mice. The results showed no statistically significant differences between the low-dose and high-dose treatment groups and the control group in white blood cell (WBC), hemoglobin (Hb), platelet (PLT), alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea, and creatinine (Cr) levels (P > 0.05). Figure 5 A, Figure 5 B).

[0063] The above experimental results show that the traditional Chinese medicine composition of the present invention has good safety in the treatment of triple-negative breast cancer-bearing mice.

[0064] Animal Experiment 2 The following is an evaluation of the effect of combining the traditional Chinese medicine composition of the present invention with PD-L1 monoclonal antibody to inhibit tumor growth and lung metastasis in triple-negative breast cancer-bearing mice.

[0065] I. Grouping, Modeling, and Drug Administration Female BALB / c mice aged 6–8 weeks, weighing 15–18 g, were housed in an SPF environment with an ambient temperature of 22–26°C and a relative humidity of 40%–60%, and were provided with sterilized food and water. After the experimental animals acclimatized to the environment, they were randomly divided into three groups: a model group, a traditional Chinese medicine composition group (ADQ807), and a group receiving a combination of traditional Chinese medicine composition and PD-L1 monoclonal antibody (ADQ807+PD-L1), with 6 mice in each group.

[0066] In the model group, the traditional Chinese medicine composition group, and the combination of traditional Chinese medicine composition and PD-L1 monoclonal antibody group, a triple-negative breast cancer cell 4T1-derived breast tumor mass was simultaneously inoculated into the left fourth pair of mammary fat pads of mice to construct a triple-negative breast cancer tumor-bearing mouse model.

[0067] When the tumor diameter reached approximately 0.5 cm, the drugs were administered to the mice in each group according to the following protocol: Traditional Chinese medicine composition group: The lyophilized powder aqueous solution (concentration of 2g / mL) of Example 5 was administered by gavage at a dose of 7.4g / kg / d once a day, with a gavage volume of 74μL; The combination of traditional Chinese medicine composition and PD-L1 monoclonal antibody: The lyophilized powder aqueous solution (concentration of 2g / mL) of Example 5 was administered by gavage at a dose of 7.4g / kg / d once a day, with a gavage volume of 74μL; at the same time, PD-L1 monoclonal antibody was injected intraperitoneally at a dose of 200μg / 20g body weight once a week.

[0068] Model group: 74 μL of normal saline was administered by gavage once a day.

[0069] Mice in each group were administered the drug by gavage for 36 consecutive days. Mice in the group treated with the combination of the traditional Chinese medicine composition and the PD-L1 monoclonal antibody were injected intraperitoneally 5 times.

[0070] The long and short axes of mouse tumors were measured every 3 days. At the end of the experiment, mice were euthanized in accordance with animal ethics requirements, and tumors and lungs were collected. The tumor weight was weighed, and the tumor and lung tissues were preserved for subsequent research.

[0071] II. Statistical Methods This study used SPSS 26.0 for statistical data analysis. Qualitative data were described by frequency and proportion; quantitative data were described by number of cases, mean ± standard deviation (if normally distributed), or median (if non-normally distributed). If the variation follows (or approximately follows) a normal distribution, a paired-samples t-test was used; if the variation does not follow a normal distribution, the Wilcoxon Signed-Rank Test was used as a robust non-parametric test. A p-value < 0.05 was considered statistically significant.

[0072] III. Test Results Statistical analysis of tumor volume and weight in mice showed that the combination of the traditional Chinese medicine composition of this invention and the PD-L1 monoclonal antibody synergistically inhibited tumor growth in triple-negative breast cancer-bearing mice (P < 0.05). Figure 6 A).

[0073] Immunohistochemical detection of Ki-67 in mouse tumor tissues showed that the combination of the traditional Chinese medicine composition of the present invention and the PD-L1 monoclonal antibody synergistically inhibited the expression of Ki-67 in tumor tissues of triple-negative breast cancer-bearing mice. Figure 6 B).

[0074] The lungs of mice were photographed, and the lung tissue was subjected to HE staining. The results showed that the traditional Chinese medicine composition of the present invention, in combination with PD-L1 monoclonal antibody, synergistically inhibited lung metastasis of tumors in triple-negative breast cancer-bearing mice. Figure 6 C Figure 6 D).

[0075] The above experimental results show that the combination of the traditional Chinese medicine composition of the present invention and the PD-L1 monoclonal antibody can synergistically inhibit tumor growth and lung metastasis in triple-negative breast cancer-bearing mice.

[0076] Typical clinical cases Typical Case 1 Ms. Li, 55 years old, underwent modified radical mastectomy for triple-negative breast cancer. Her symptoms included fatigue, dizziness, palpitations, aversion to cold, cold limbs, poor sleep with vivid dreams, pale red tongue with a thin white coating, and weak pulse. Her Piper Fatigue Scale score was 4.77, and her FACT-B score was 81. After taking the herbal granules of Example 3 of this invention for two weeks, two sachets (13.5g each) daily, taken in the morning and evening, her symptoms improved and her condition improved. Her Piper Fatigue Scale score was 3.5, and her FACT-B score was 103.

[0077] This patient underwent surgery for triple-negative breast cancer. She had completed eight cycles of neoadjuvant chemotherapy before surgery and adjuvant radiotherapy after surgery. She currently experiences fatigue, dizziness, blurred vision, and palpitations. Based on the four diagnostic methods (inspection, auscultation and olfaction), she was diagnosed with a deficiency of both Qi and Blood. Previous surgery and chemotherapy depleted her Qi and Blood. Since Qi is the commander of Blood and Blood is the mother of Qi, the patient's current condition is primarily due to Qi deficiency. Treatment should follow the principle of tonifying Qi and generating Blood. Therefore, this formula heavily uses Astragalus membranaceus to tonify Qi and raise Yang, combined with Curcuma zedoaria and Hedyotis diffusa for detoxification and anti-cancer effects. This not only significantly alleviated the patient's fatigue symptoms and improved her quality of life but also helped prevent recurrence and metastasis of triple-negative breast cancer.

[0078] Typical Case 2 Ms. Kuang, female, 56 years old, post-operative triple-negative breast cancer, during capecitabine intensive therapy, presented with fatigue, shortness of breath upon exertion, spontaneous sweating, night sweats, dizziness, palpitations, pale tongue with a thin white coating, and weak pulse. Auxiliary examination: Neutrophils: 1.78 × 10⁻⁶ 9 / L. The average score on the Piper Fatigue Revision Scale was 3.48, and the total score on the FACT-B scale was 83. After taking the traditional Chinese medicine granules of Example 3 of this invention for 2 weeks, 2 bags daily, each bag containing 13.5g, taken in the morning and evening, the symptoms improved and the condition was relieved. The average score on the Piper Fatigue Revision Scale was 1.9, and the FACT-B scale score was 100.

[0079] This patient, undergoing chemotherapy after surgery for triple-negative breast cancer, experienced fatigue, shortness of breath upon exertion, spontaneous sweating, night sweats, dizziness, palpitations, pale tongue with a thin white coating, and weak pulse during capecitabine intensive therapy. These symptoms were indicative of qi and blood deficiency, accompanied by bone marrow suppression symptoms such as decreased neutrophils, suggesting an adverse reaction to capecitabine. Chemotherapy drugs deplete the spleen and qi, leading to insufficient acquired constitution and, over time, damage to congenital essence and blood, resulting in bone marrow suppression. Since tangible blood cannot be quickly generated, and intangible qi must be urgently consolidated, a large dose of Astragalus membranaceus was used to consolidate intangible qi. Qi generates blood, and Paeonia lactiflora was added to replenish blood deficiency. Combined with Curcuma zedoaria and Hedyotis diffusa to remove blood stasis, detoxify, and fight cancer, the patient's fatigue symptoms were alleviated, and the treatment played a role in reducing toxicity and enhancing efficacy during capecitabine intensive therapy.

Claims

1. A traditional Chinese medicine composition for treating triple-negative breast cancer, characterized in that, Its main active ingredients, by weight, include: Astragalus membranaceus 20-40 parts, Curcuma zedoaria 10-20 parts, Hedyotis diffusa 10-20 parts, Citrus aurantium 5-15 parts, Bupleurum chinense 5-15 parts, Paeonia lactiflora 5-15 parts, and Glycyrrhiza uralensis 5-15 parts.

2. The traditional Chinese medicine composition for treating triple-negative breast cancer according to claim 1, characterized in that, Its main active ingredients, by weight, include: Astragalus membranaceus 30 parts, Curcuma zedoaria 15 parts, Hedyotis diffusa 15 parts, Citrus aurantium 10 parts, Bupleurum chinense 10 parts, Paeonia lactiflora 10 parts, and Glycyrrhiza uralensis 10 parts.

3. The traditional Chinese medicine composition for treating triple-negative breast cancer according to claim 1 or 2, characterized in that, The licorice mentioned is roasted licorice.

4. The traditional Chinese medicine composition for treating triple-negative breast cancer according to claim 1 or 2, characterized in that, It also includes pharmaceutically acceptable excipients.

5. The traditional Chinese medicine composition for treating triple-negative breast cancer according to claim 4, characterized in that, The excipients are at least one of the following: fillers, suspending agents, disintegrants, lubricants, adhesives, preservatives, sweeteners, and flavoring agents.

6. The traditional Chinese medicine composition for treating triple-negative breast cancer according to claim 1 or 2, characterized in that, The dosage forms of the traditional Chinese medicine composition are decoctions, oral solutions, mixtures, decoction pastes, granules, freeze-dried powders, powders, tablets, pills, capsules, sustained-release preparations, controlled-release preparations, ointments, or suppositories.

7. The use of the traditional Chinese medicine composition for treating triple-negative breast cancer according to any one of claims 1 to 6 in the preparation of a medicament for treating triple-negative breast cancer.

8. The use of the traditional Chinese medicine composition for treating triple-negative breast cancer according to any one of claims 1 to 6 in the preparation of a medicament that enhances the efficacy of PD-L1 monoclonal antibody in inhibiting tumor growth and lung metastasis.

9. The application according to claim 8, characterized in that, The tumor is a triple-negative breast cancer tumor.

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