Traditional Chinese medicine composition for treating breast cancer as well as preparation and application thereof

Abstract

The invention discloses a traditional Chinese medicine composition for treating breast cancer and a preparation and application thereof, and belongs to the technical field of traditional Chinese medicine and network pharmacology. The traditional Chinese medicine composition comprises the following raw material medicines: radix bupleuri, angelica sinensis, radix paeoniae alba, radix curcumae, dandelion, uniflower swisscentaury root, sculellaria barbata, selfheal, rhizoma bolbostemmae, thunberg fritillary bulb, spina gleditsiae, turtle shell, salvia miltiorrhiza and glossy privet fruit. The main effective components of the traditional Chinese medicine composition are quercetin, luteolin, kaempferol, beta-sitosterol and the like, and can participate in conduction of Hippo, TGF-beta, axon guidance, tight junction, vascular smooth muscle contraction and other signal channels through PTGS2, PTGS1, NCOA2, ADRB2 and other targets. Further, the anti-tumor effect is achieved in the biological processes of positively regulating cell growth, negatively regulating and controlling an external apoptosis signal path through a death structural domain receptor, positively regulating an endothelial cell apoptosis process, reaction to cytokines, an internal apoptosis signal path, a hormone-mediated signal transduction path and the like.

Description

Technical Field

[0001] This invention relates to the fields of traditional Chinese medicine and network pharmacology, specifically to a traditional Chinese medicine composition for treating breast cancer, its preparation, and its application. Background Technology

[0002] According to global cancer statistics in 2020, breast cancer ranks second in incidence among 36 cancers in 185 countries surveyed worldwide, at 11.6%, and fourth in mortality at 6.9%, making it the most common cancer among women. In my country, in 2022, breast cancer accounted for 11.5% of all cancer cases, ranking fourth, and 23.8% of all female cancer cases, ranking second. Its mortality rate accounted for 6.8% of all cancer deaths and 15.4% of all female cancer cases, ranking fifth. Breast cancer is a heterogeneous disease with a unique epidemiological pattern. Currently, Western medicine mainly uses surgery, chemotherapy, radiotherapy, endocrine therapy, and targeted therapy to treat breast cancer, but these methods often lead to a series of side effects. Especially for triple-negative breast cancer, endocrine and targeted therapies are ineffective, and chemotherapy, radiotherapy, and surgery are common treatments. Therefore, researching and leveraging the unique advantages of Traditional Chinese Medicine (TCM) in inhibiting breast cancer cell proliferation and metastasis, reducing the toxic side effects of chemotherapy, and improving patients' quality of life is particularly important.

[0003] In traditional Chinese medicine, there is no record of the disease name "breast cancer". Chen Ziming, a medical expert in the Southern Song Dynasty, had a basic understanding of this disease in "Gynecological Prescriptions for Women of All Ages", named it "breast carcinoma", and proposed that its etiology and pathogenesis were liver and spleen depression and anger, as well as qi and blood deficiency. In "Orthodox Manual of Surgery" by Chen Shigong, from the perspective of the association between the five emotions and the five zang-organs, it was proposed that the etiology and pathogenesis of breast cancer were melancholy injuring the liver, overthinking injuring the spleen, meridian obstruction, and accumulation into nodules. Zhu Zhenheng proposed in "Discourses on Investigation and Extension" that "the breast is related to the meridian of Yangming, and the nipple belongs to Jueyin", clarifying the internal connection between breast cancer and the liver and spleen from the perspective of meridians. Zheng Wanqun et al.'s research on the dialectical classification of breast cancer in traditional Chinese medicine divided it into qi and blood deficiency syndrome, liver and kidney yin deficiency syndrome, thoroughfare and conception vessel disharmony, and liver depression and phlegm coagulation syndrome; Li Dehui et al. studied the correlation between the traditional Chinese medicine syndrome types of breast cancer and TNM staging, and classified the included cases according to traditional Chinese medicine syndrome types. The most common type was liver depression and phlegm coagulation type, followed by thoroughfare and conception vessel disharmony type and healthy qi deficiency and toxin flaming type in turn. Ma Yunfei et al. analyzed and summarized the relevant literature on the traditional Chinese medicine syndromes of breast cancer from three aspects: the dialectical experience of famous doctors, the macroscopic dialectical rules of statistics, and the basic microscopic dialectical research, and found that the traditional Chinese medicine syndromes of breast cancer were mainly related to the liver and spleen in the early stage, often liver depression and spleen deficiency type, and in the later stage, there would be spleen and kidney deficiency and liver and kidney deficiency. Surgery, radiotherapy and chemotherapy were likely to lead to qi and blood deficiency and qi and yin deficiency. Looking at the understanding of breast cancer by ancient and modern medical experts, it is believed that the onset of breast cancer is related to the liver, spleen and kidney, the nature of the disease is deficiency and excess mixed, the basic pathogenesis is healthy qi deficiency, disharmony of zang-fu organs and yin and yang, liver depression and spleen deficiency, abnormal dispersion and discharge, phlegm coagulation and blood stasis, accumulation into nodules, and development into breast carcinoma. The treatment method should be soothing the liver and relieving depression, strengthening the spleen and tonifying the kidney, resolving phlegm and removing blood stasis, and detoxifying and dissipating nodules.

[0004] Network pharmacology integrates systems pharmacology, biology, computer science and other disciplines. By using the means of network pharmacology, it can reveal the action principle of traditional Chinese medicine by constructing a complex network interaction diagram among the active ingredients of traditional Chinese medicine, disease targets and pathways, and verify the results obtained by network pharmacology by means of molecular docking, showing great advantages at the molecular level of studying the treatment of diseases by traditional Chinese medicine. Summary of the Invention

[0005] Based on this, it is necessary to provide a traditional Chinese medicine composition for treating breast cancer, its preparation and application in view of the problems such as the unclear action mechanism of the traditional Chinese medicine formula currently used for treating breast cancer and the lack of effective demonstration of the action effect.

[0006] The present invention provides a traditional Chinese medicine composition for treating breast cancer, and the traditional Chinese medicine composition comprises the following raw materials: bupleurum, angelica sinensis, white peony root, turmeric, dandelion, rhaponticum uniflorum, scutellaria barbata, prunella vulgaris, bolbostemma paniculatum, fritillaria thunbergii, gleditsia spine, turtle shell, salvia miltiorrhiza, glossy privet fruit.

[0007] Preferably, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: Bupleurum chinense 10-15 parts, Angelica sinensis 8-12 parts, Paeonia lactiflora 8-12 parts, Curcuma longa 8-12 parts, Taraxacum mongolicum 6-9 parts, Echinops latifolius 6-9 parts, Scutellaria barbata 6-9 parts, Prunella vulgaris 6-9 parts, Fritillaria thunbergii 3-6 parts, Fritillaria thunbergii 10-12 parts, Gleditsia sinensis 6-9 parts, Trionyx sinensis 6-9 parts, Salvia miltiorrhiza 6-12 parts, and Ligustrum lucidum 10-15 parts.

[0008] Preferably, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 15 parts Bupleurum, 12 parts Angelica sinensis, 12 parts Paeonia lactiflora, 9 parts Curcuma longa, 9 parts Taraxacum mongolicum, 9 parts Echinops latifolius, 9 parts Scutellaria barbata, 9 parts Prunella vulgaris, 6 parts Fritillaria thunbergii, 12 parts Fritillaria thunbergii, 9 parts Gleditsia sinensis thorns, 9 parts Trionyx sinensis, 12 parts Salvia miltiorrhiza, and 15 parts Ligustrum lucidum.

[0009] The present invention also provides a traditional Chinese medicine preparation for treating breast cancer, which is prepared from the above-mentioned traditional Chinese medicine composition for treating breast cancer.

[0010] Preferably, the traditional Chinese medicine preparation is a decoction, which is prepared according to the following steps:

[0011] (1) Weigh the raw materials according to the weight parts, add 5-10 times the weight parts of water and soak for 30-120 minutes;

[0012] (2) Heat and boil the soaked raw materials together with water, filter, and collect the filtrate;

[0013] (3) Add 5-10 times the weight of water to the filter residue, boil, filter, and collect the filtrate;

[0014] (4) Combine the filtrates obtained after two decoctions and heat them to concentrate, and the decoction is obtained.

[0015] Preferably, the decocting time for both times is 30-60 minutes.

[0016] Preferably, the filtrates are combined and then concentrated by heating to 10%-20% of their original volume.

[0017] Furthermore, the present invention also provides the application of the above-mentioned traditional Chinese medicine composition for treating breast cancer in the preparation of a medicament for treating breast cancer.

[0018] The formula for the traditional Chinese medicine composition for treating breast cancer in this invention is explained as follows:

[0019] The primary cause of breast cancer is attributed to the liver, and its treatment should prioritize regulating the liver. The formula uses Bupleurum, Angelica sinensis, Paeonia lactiflora, and Curcuma longa as the principal herbs. Bupleurum and Curcuma longa soothe the liver and regulate Qi, while Angelica sinensis and Paeonia lactiflora nourish and invigorate blood, soften the liver, and relieve pain. Dandelion, Echinops latifolius, and Scutellaria barbata clear heat and detoxify, reduce swelling and dissipate nodules; together with Prunella vulgaris, they enter the liver meridian to clear liver fire and treat mastitis and breast lumps. Fritillaria thunbergii, Fritillaria thunbergii, and Gleditsia sinensis thorns dissipate nodules and reduce swelling. Turtle shell, a substance rich in blood and flesh, softens and dissipates nodules and nourishes the liver and kidneys. These eight herbs serve as assistant herbs, primarily focusing on dissipating nodules and reducing swelling to enhance efficacy. Salvia miltiorrhiza is known for its ability to invigorate blood and remove blood stasis, while Ligustrum lucidum nourishes the liver and kidneys. All 14 herbs work together to soothe the liver, relieve depression, regulate Qi, invigorate blood, relieve pain, reduce swelling, and dissipate nodules to treat breast cancer.

[0020] Specifically, each of the raw materials for traditional Chinese medicine has the following pharmacological characteristics:

[0021] Bupleurum: This product is the dried root of Bupleurum chinense DC. or Bupleurum corzonerifolium Willd., both belonging to the Apiaceae family. It has a pungent, bitter, and slightly cold taste. It enters the liver, gallbladder, and lung meridians. It has the effects of dispersing and reducing fever, soothing the liver and relieving depression, and raising yang qi.

[0022] Angelica sinensis: This product is the dried root of Angelica sinensis (Oliv.) Diels, a plant belonging to the Apiaceae family. It has a sweet and pungent taste, and is warm in nature. It enters the liver, heart, and spleen meridians. It has the effects of nourishing blood and promoting blood circulation, regulating menstruation and relieving pain, and moistening the intestines and promoting bowel movements.

[0023] White peony root: This product is the dried root of Paeonia lactiflora Pall., a plant in the Ranunculaceae family. It is bitter, sour, and slightly cold in nature. It enters the liver and spleen meridians. It nourishes blood and regulates menstruation, astringes yin and stops sweating, softens the liver and relieves pain, and calms liver yang.

[0024] Turmeric: This product is the dried tuberous root of *Curcuma wenyujin* YHChen et C.Ling, *Curcuma longa* L., *Curcuma kwangsiensis* SGLee et CFLiang, or *Curcuma phaeocaulis* Val. (all belonging to the ginger family). It has a pungent, bitter, and cold nature. It enters the liver, heart, and lung meridians. It has the effects of promoting blood circulation and relieving pain, regulating qi and relieving depression, clearing the heart and cooling the blood, and promoting bile secretion and reducing jaundice.

[0025] Dandelion: This product is the dried whole herb of *Taraxacum mongolicum* Hand.-Mazz., *Taraxacum borealisinense* Kitam., or several other species of the same genus in the Asteraceae family. It tastes bitter and sweet, and is cold in nature. It enters the liver and stomach meridians. It has the effects of clearing heat and detoxifying, reducing swelling and dissipating nodules, and promoting urination.

[0026] Rhaponticum uniforum (L.) DC., a plant in the Asteraceae family, is a dried root. It is bitter and cold in nature, and enters the Stomach meridian. It has the effects of clearing heat and detoxifying, reducing swelling, promoting lactation, and relaxing muscles and tendons.

[0027] Scutellaria barbata D. Don, a plant in the Lamiaceae family, is a dried whole herb. It has a pungent, bitter, and cold nature. It enters the lung, liver, and kidney meridians. It has the effects of clearing heat and detoxifying, resolving blood stasis, and promoting diuresis.

[0028] Prunella vulgaris: This product is the dried fruit spike of Prunella vulgaris L., a plant of the Lamiaceae family. It has a pungent, bitter, and cold nature. It enters the liver and gallbladder meridians. It has the effects of clearing liver heat, improving eyesight, and dispersing nodules and reducing swelling.

[0029] Fritillaria paniculata: This product is the dried tuber of *Bolbostemma paniculatum* (Maxim.) Franquet, a plant in the Cucurbitaceae family. It has a bitter taste and is slightly cold in nature. It enters the lung and spleen meridians. It has detoxifying, nodule-dispersing, and swelling-reducing effects.

[0030] Fritillaria thunbergii Miq. (Zhejiang Fritillaria): This product is the dried bulb of Fritillaria thunbergii Miq., a plant belonging to the Liliaceae family. It is bitter and cold in nature. It enters the Lung and Heart meridians. It has the effects of clearing heat, resolving phlegm, relieving cough, detoxifying, dispersing nodules, and reducing swelling.

[0031] Gleditsia sinensis thorns: This product is the dried thorns of the legume Gleditsia sinensis Lam. It has a pungent and warm nature. It enters the liver and stomach meridians. It has the effects of reducing swelling and promoting pus drainage, and killing parasites.

[0032] Turtle shell: This product is the carapace of the turtle Trionyx sinensis Wiegmann, a species of turtle in the family Trionychidae. It has a salty taste and is slightly cold in nature. It enters the liver and kidney meridians. It has the effects of nourishing yin and suppressing yang, reducing fever and eliminating steaming sensations, and softening hard masses and dispersing nodules.

[0033] Danshen: This product is the dried root and rhizome of *Salvia miltiorrhiza* Bge., a plant belonging to the Lamiaceae family. It has a bitter taste and is slightly cold in nature. It enters the Heart and Liver meridians. It has the effects of promoting blood circulation and removing blood stasis, regulating menstruation and relieving pain, clearing the heart and relieving irritability, cooling the blood and reducing swelling.

[0034] Privet fruit: This product is the dried, ripe fruit of *Ligustrum lucidum* Ait., a plant in the Oleaceae family. It has a sweet and bitter taste, and is cool in nature. It enters the liver and kidney meridians. It nourishes the liver and kidneys, improves eyesight, and darkens hair.

[0035] The mechanism of action, targets, and pathways of the traditional Chinese medicine composition for treating breast cancer of this invention, as determined by network pharmacology studies, are as follows:

[0036] By screening the main components of the traditional Chinese medicine composition of this invention and constructing a schematic diagram of the multi-level interaction network between components and targets, the active components were found to be quercetin, luteolin, kaempferol, β-sitosterol, artemisinin, etc. Quercetin is one of the most common flavonoids, possessing anti-inflammatory and anti-tumor properties. It can inhibit tumor angiogenesis by targeting the vascular endothelial growth factor (VEGF) and vitamin D pathways, and can control cell apoptosis and inhibit migration and proliferation. In addition, quercetin is also a natural immunomodulator that can inhibit anaerobic glycolysis in tumor cells, reduce lactic acid production and transport, reduce the acidification of the tumor microenvironment, reverse immunosuppression, enhance the activity of immune cells, and directly regulate the tumor local microenvironment (TME). Furthermore, quercetin has also been shown to be a synergist, which can further enhance the anticancer activity of commonly used antitumor agents, thereby reducing their dosage, and can also resensitize drug-resistant cancer cells. Luteolin possesses antitumor, lipid-lowering, and antioxidant effects. Its monoacylated derivatives exhibit higher bioavailability, resulting in better anti-proliferative and antioxidant activity against MDA-MB-231 breast cancer. Luteolin can inhibit the progression of the cell cycle in the G1 and sub-G1 phases of MCF-7 breast cancer cells. It can induce cell cycle arrest in the S phase by reducing telomerase levels and inhibiting phosphorylation of NF-κB inhibitor α in MDA-MB-231 cells. It can also inhibit poly-ADP-ribose polymerase, which helps cancer cells repair themselves. Kaempferol's anti-breast cancer mechanism mainly involves inducing DNA damage. It induces apoptosis through Nrf2 regulation of ROS production, increased PARP and Bax cleavage, downregulation of Bcl-2 expression, and increased expression of caspase 3, caspase 9, and pAMT. It induces cell cycle arrest in the G2 / M phase, inhibits immune evasion by regulating the JAK-STAT 3 pathway, and inhibits tumor angiogenesis and metastatic potential by downregulating MMP-3 and MMP-9 levels. β-Sitosterol stimulates apoptosis in MDA-MB-231 breast cancer cells by increasing the Bax / Bcl-2 ratio and promoting mitochondrial membrane depolarization. It inhibits the migration of MCF-7 and MDA-MB-231 cells in human breast cancer by upregulating CDH1 expression. High concentrations of β-sitosterol can also inhibit human breast cancer T47D cells from entering the G0 / G1 phase, thereby reducing the proportion of cells in the S phase and lowering the proliferation index, thus inhibiting cell proliferation. Furthermore, β-sitosterol exhibits estrogen-like effects within a certain concentration range, exerting these effects by regulating the expression of estrogen-inducible protein 2 (EMP-2) gene via the estrogen receptor pathway. Artemisinin, at doses of 10-50 μM, significantly reduces the survival rate of HeLa cancer cells and regulates their lipid distribution. It also exhibits significant cytotoxic activity against human breast cancer cells (MCF-7 and T47D), regulating microtubule depolymerization and leading to apoptosis by activating the mitotic spindle gate.

[0037] The main targets of the herbal composition for treating breast cancer (BC) in this invention are PTGS2, PTGS1, NCOA2, ADRB2, and SCN5A. Prostaglandin endoperoxidase 2 (PTGS2), also known as cyclooxygenase-2 (COX-2), participates in cell homeostasis, angiogenesis, and intercellular signaling. Overexpression of COX-2 is associated with increased prostaglandin E2 (PGE2) levels, and high PGE2 levels are linked to breast cancer development. Inhibiting NCOA2 gene expression can induce cell cycle arrest and apoptosis, significantly inhibiting cell proliferation in different breast cancer cell lines. The specific mechanism may be through downregulation of its downstream target RASEF, leading to downregulation of the MAPK / ERK signaling cascade. Studies have shown that ADRB2 knockdown can reduce the proliferative activity of TNBC cells, highlighting its potential value as a target for improving chemosensitivity. The gene encoding the SCN5A voltage-gated sodium ion channel NaV1.5 can promote cell proliferation, colony formation, movement / migration and invasion. Targeted therapy of NaV1.5 / naV1.5 can inhibit tumor growth and lung metastasis in an orthotopic TNBC model. The targeted expression of NaV1.5 / naV1.5 is related to the expression of eukaryotic cell extension factor-2 kinase (EF2K).

[0038] The study identified pathways involved in the development and progression of breast cancer, primarily through tumorigenesis and proliferation, metastasis and invasion, metabolic and microenvironment regulation, immunity and inflammation, and treatment resistance. Among these, the Hippo signaling pathway and the TGF-β signaling pathway likely play crucial roles. The Hippo signaling pathway is closely related to cell proliferation, migration, invasion, metastasis, and treatment resistance in breast cancer. The transcriptional coactivator Yes-associated protein (YAP) / PDZ-binding transcriptional coactivator (TAZ) is an upstream factor in the Hippo signaling pathway. The complex formed by the interaction of YAP / TAZ with transcription factors of the transcriptional enhancer domain (TEAD) family, especially TEAD4, activates the transcription of downstream target genes, promoting the proliferation and invasion of breast cancer cells. The TGF-β signaling pathway has a dual role, exerting protective or tumor-suppressive effects in the early growth-sensitive stages of tumor development. In the late stages of malignant tumors, TGF-β overload drives tumor progression. TGF-β1 and BMP-7 (bone morphogenetic protein-7) are the main members of the TGF-β receptor ligand family. Studies have found that TGF-β1 signaling is overactivated in breast cancer, and the absence of BMP-7 in breast cancer is prone to lead to enhanced EMT and the acquisition of invasive / metastatic features.

[0039] Compared with the prior art, the present invention has the following advantages:

[0040] The herbal composition of this invention focuses on the principles of soothing the liver and relieving depression, strengthening the spleen and tonifying the kidneys, resolving phlegm and removing blood stasis, and detoxifying and dispersing nodules. Bupleurum and Curcuma soothe the liver and regulate qi; Curcuma also invigorates blood and relieves pain. Angelica and Paeonia lactiflora nourish blood and astringe yin; Angelica, with its pungent and dispersing flavor, is a qi-regulating herb in the blood, capable of both tonifying and invigorating, aligning with the nature of the liver. The four herbs combined nourish the liver and assist its function, resulting in smooth blood and harmonious liver function. Breast cancer is a nodule formed by the mutual binding of phlegm, blood stasis, and toxins; targeting this pathological product, Fritillaria thunbergii and Fritillaria cirrhosa are added. The mother herb resolves phlegm and dissipates nodules; Fritillaria cirrhosa also has detoxifying properties and is effective in treating scrofula and phlegm nodules. Prunella vulgaris, Taraxacum mongolicum, and Scutellaria barbata are added to clear heat, detoxify, and dissipate nodules. Scutellaria barbata is also a classic anti-cancer drug. Echinops latifolius and Gleditsia sinensis are further added to clear heat, detoxify, reduce swelling, and dissipate nodules. Echinops latifolius can also promote menstruation and lactation, while Gleditsia sinensis enters the liver and stomach meridians. Both guide the medicine directly to the affected area. Finally, Salvia miltiorrhiza, Ligustrum lucidum, and Trionyx sinensis are added to invigorate qi and blood, and tonify the liver and kidneys. Trionyx sinensis is particularly effective in softening hardness and dissipating nodules. The combined effects of these herbs treat both the root cause and the symptoms, combining attack and tonification, and regulating the liver, spleen, and kidneys. Together, they achieve the effects of soothing the liver and strengthening the spleen, resolving phlegm and removing blood stasis, and detoxifying and dissipating nodules.

[0041] The main effective components of the traditional Chinese medicine composition for treating breast cancer in this invention are quercetin, luteolin, kaempferol, β-sitosterol, and artemisinin; the targets include PTGS2, PTGS1, NCOA2, ADRB2, and SCN5A; it may be related to signaling pathways such as the Hippo signaling pathway, TGF-β signaling pathway, axonal guidance, tight junctions, vascular smooth muscle contraction, cGMP-PKG signaling pathway, and protein processing in the endoplasmic reticulum; the biological processes it may participate in mainly include actively regulating cell growth, negatively regulating external apoptosis signaling pathways through death domain receptors, actively regulating endothelial cell apoptosis, responses to cytokines, intrinsic apoptosis signaling pathways, and hormone-mediated signal transduction pathways; molecular docking verification also shows that the binding energy of most active components is less than -7.0 kcal / mol, and most targets and components have good binding activity and tight binding. Therefore, the traditional Chinese medicine composition of this invention mainly participates in the treatment of breast cancer through multiple targets and pathways, with a clear mechanism of action, and provides a theoretical basis for clinical application and new drug development through network pharmacology research. Attached Figure Description

[0042] Figure 1 Venn diagram showing the drug targets and breast cancer disease gene targets of the traditional Chinese medicine composition for treating breast cancer in the embodiments of the present invention;

[0043] Figure 2 This is a PPI network diagram of the intersection genes of the pharmaceutical compositions in the embodiments of the present invention;

[0044] Figure 3 This is a gene diagram of an important functional module 1 for the treatment of breast cancer using the traditional Chinese medicine composition in this embodiment of the invention;

[0045] Figure 4 This is a gene diagram of an important functional module 2 in the treatment of breast cancer using the traditional Chinese medicine composition in this invention.

[0046] Figure 5 This is a gene diagram of three important functional modules in the treatment of breast cancer using the traditional Chinese medicine composition in this invention.

[0047] Figure 6 This is a GO-BP analysis diagram of the gene module of the traditional Chinese medicine composition in the embodiments of the present invention;

[0048] Figure 7 This is a KEGG analysis diagram of the gene module of the traditional Chinese medicine composition in the embodiments of the present invention;

[0049] Figure 8 This is a GO analysis diagram of the intersection genes of the herbal compositions in the embodiments of the present invention;

[0050] Figure 9 This is a KEGG analysis diagram of the intersection genes of the herbal compositions in the embodiments of the present invention;

[0051] Figure 10 This is a network diagram of the effective components, intersection genes, and pathways of the traditional Chinese medicine composition in the embodiments of the present invention.

[0052] Figure 11 This is a diagram illustrating the docking structure between the active ingredient of the traditional Chinese medicine composition and the core target molecule in an embodiment of the present invention. Detailed Implementation

[0053] To make the above-mentioned objects, features, and advantages of the present invention more apparent and understandable, specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings. Many specific details are set forth in the following description to provide a thorough understanding of the present invention. However, the present invention can be practiced in many other ways different from those described herein, and those skilled in the art can make similar modifications without departing from the spirit of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0054] Example 1

[0055] Screening of effective components in traditional Chinese medicine compositions for treating breast cancer

[0056] The main compounds of Bupleurum chinense, Angelica sinensis, Paeonia lactiflora, Curcuma longa, Echinops latifolius, Fritillaria thunbergii, Gleditsia sinensis, Prunella vulgaris, Fritillaria cirrhosa, Scutellaria barbata, Salvia miltiorrhiza, and Ligustrum lucidum were retrieved using the Traditional Chinese Medicine Systems Pharmacology Database (TCMSP, http: / / lsp.Nwsuaf.edu.cn / tcmsp.php). Compounds with OB ≥ 30% and DL ≥ 0.18 were retained. Effective components not found in the TCMSP database were supplemented through literature mining. Furthermore, for drugs not found in the TCMSP database, Carapax Trionycis and Taraxacum mongolicum, Herb database, ETCM database, and literature were used as supplementary sources.

[0057] Preliminary extraction yielded 20 effective components from Bupleurum chinense, 2 from Angelica sinensis, 13 from Paeonia lactiflora, 15 from Curcuma longa, 5 from Echinops latifolius, 7 from Fritillaria thunbergii, 11 from Gleditsia sinensis thorns, 11 from Prunella vulgaris, 13 from Fritillaria cirrhosa, 29 from Scutellaria barbata, 65 from Salvia miltiorrhiza, 13 from Ligustrum lucidum, 2 from Carapax Trionycis, and 34 from Taraxacum mongolicum (see Table 1).

[0058] Table 1. Main effective components of traditional Chinese medicine compositions for treating breast cancer.

[0059]

[0060]

[0061]

[0062]

[0063] Example 2

[0064] Screening of active ingredient targets, disease target screening, and prediction of potential targets for treating breast cancer using traditional Chinese medicine compositions.

[0065] Targets of active ingredients were collected using the TCMSP database. For active ingredients without targets in the TCMSP database, target prediction was performed using the ETCM, Herb, and Swiss Target databases. SMILES structures of active ingredients were retrieved from the PubChem database, and corresponding targets were retrieved from the Swiss Target Prediction database, retaining targets with a probability greater than or equal to 0.1. In the GeneCards, OMMIM, and TTD databases, disease genes for breast cancer (BC) were retrieved using "breast cancer" as the criterion, with "Homo spaiens" as the species selection. Disease genes from the three databases were merged and duplicated; higher score values ​​indicate a higher correlation between the gene and the disease. If there were many disease targets, targets with scores greater than the median, mean, or quartile could be designated as potential targets for BC. The combination drug targets and disease genes were imported into Venny 2.1 for Venn analysis to screen for potential therapeutic targets for BC, i.e., the intersection genes between the combination drug targets and disease genes.

[0066] 256 effective target molecules were identified in Bupleurum chinense, 36 in Angelica sinensis, 102 in Paeonia lactiflora, 92 in Curcuma longa, 31 in Echinops latifolius, 131 in Fritillaria thunbergii, 211 in Gleditsia sinensis, 160 in Prunella vulgaris, 29 in Fritillaria cirrhosa, 187 in Scutellaria barbata, 225 in Salvia miltiorrhiza, 156 in Ligustrum lucidum, 12 in Carapax Trionycis, and 300 in Taraxacum mongolicum. After integration and deduplication, the protein targets of the compounds were converted into gene names using the Uiprot protein database, retaining a total of 959 targets. 18,253 BC targets were retrieved from the GeneCards database, 20 BC targets were obtained from the OMMIM database, and 101 BC targets were obtained from the TTD database. After merging and deduplicating, 18,253 disease targets were obtained. Since there were still many disease targets with scores higher than the average after taking the average, genes with a score ≥ 15 were selected, and 2,315 disease genes were screened out.

[0067] Example 3

[0068] Construction and functional module identification of drug intersection gene PPI network for traditional Chinese medicine composition in breast cancer treatment

[0069] To clarify the interactions between potential therapeutic targets in combination therapy for BC, the intersecting genes obtained from Wayne analysis were imported into the STRING database, and the biological species were set as "Homosapiens". PPI network diagrams and tsv files of the PPI networks were obtained. The tsv files were imported into CytoScape software, and the Mcode plugin in CytoScape was used to identify functional modules of the network of intersecting gene interactions. The three functional module networks with a score >6 were retained and visualized.

[0070] 959 compound drug targets and 2315 disease genes were introduced into Venny 2.1 for Venn analysis, yielding 318 intersection genes between the compound drug targets and disease genes. (See...) Figure 1 The STRING database was used to analyze the protein-protein interactions of the intersecting genes, resulting in a PPI network diagram, see [link to diagram]. Figure 2 The Mcode plugin within CytoScape software was used to further identify functional modules in the PPI network. A total of 8 functional modules were identified. Three functional module networks with a Score > 6 were selected for visualization. (See attached image.) Figure 3-5 .

[0071] Example 4

[0072] Enrichment analysis of potential target functions and pathways related to the treatment of breast cancer using traditional Chinese medicine compositions.

[0073] The David database, based on bioinformatics techniques, uses statistical methods to perform enrichment analysis of target genes' functions and pathways, revealing their mechanisms of action in disease occurrence and development. Target functions mainly include important biological processes (BP), cellular components (CC), and molecular functions (MF) involved by the target genes or proteins. To further analyze the biological processes involved by potential protein functional modules, the genes and proteins of three functional modules were imported into the David database (https: / / david.ncifcrf.gov / ), with an FDR of <0.05, for enrichment analysis of their main biological processes and metabolic pathways. The analysis results were saved and imported into the MicroBioinformatics platform (http: / / www.bioinformatics.com.cn) for visualization. Simultaneously, the David database was used to perform enrichment analysis and visualization of target functions and pathways on intersecting genes, comparing the enrichment results of functional modules and intersecting genes.

[0074] Three groups of functional module gene proteins with a score > 6 were imported into the David database, and enrichment analysis was performed on their main biological processes and metabolic pathways using FDR < 0.05 as the screening criterion. Module 1 had the highest score of 59.632, making it the main functional module. It yielded the most enriched entries, totaling 111, including 181 BPs and 33 pathways. Module 1 was primarily enriched in biological processes involving mitochondrial release of cytochrome c and positive regulation of mesenchymal cell proliferation, mainly in the melanin production pathway. Module 2 had a score of 20.29, yielding 81 enriched entries, including 56 BPs and 25 pathways. Module 2 was primarily enriched in biological processes involving positive regulation of the ERK1 and ERK2 cascades and the Fc-gamma receptor signaling pathway involved in phagocytosis, mainly in the Kaposi's sarcoma-associated herpesvirus infection pathway. Module 3 had a score of 6.558, yielding 67 enriched entries, including 39 BPs and 28 pathways. Module 3 was primarily enriched in biological processes involving cytokine-mediated signaling pathways, mainly in the progesterone-mediated oocyte maturation pathway. Visualize the top 10 enrichment results with the lowest FDR values ​​from each of the three modules, see [link / reference]. Figure 6-7 Enrichment results indicate that the compound drug may achieve its therapeutic effect on BC by regulating certain targets in the above-mentioned biological processes.

[0075] 318 potential therapeutic targets were imported into the DAVID database for GO function analysis. Using FDR < 0.05 as the screening criterion, 484 entries were obtained (374 BP, 34 CC, and 76 MF). Based on the FDR values, the top 10 BP, CC, and MF data were imported into the MicroBioinformatics platform and analyzed as bubble charts. The results are shown below. Figure 8-9 In terms of biological processes (BP), this involves the active regulation of cell growth, negative regulation of external apoptosis signaling pathways through death domain receptors, active regulation of endothelial cell apoptosis, responses to cytokines, intrinsic apoptosis signaling pathways, hormone-mediated signal transduction pathways, osteoclast differentiation, proteolysis, dendritic cell chemotaxis, and microglia activation. In terms of cellular components (CC), this involves the cytoplasm, cell, nucleoplasm, core, protein complexes, receptor complexes, chromatin, perinuclear region, extracellular space, and membrane rafts. In terms of molecular functions (MF), this involves enzyme binding, protein kinase activity, histone H2AXY142 kinase activity, histone H3Y41 kinase activity, protein tyrosine kinase activity, ATP binding, protein serine / threonine kinase activity, same-protein binding, protein serine kinase activity, and nuclear receptor activity.

[0076] KEGG pathway analysis was performed on 318 targets, and 28 pathways were identified with an FDR < 0.05. These pathways involved axonal guidance, vascular smooth muscle contraction, cell membrane DNA sensing pathways, hematopoietic cell lines, viral myocarditis, tight junctions, arachidonic acid metabolism, long-term synergistic effects, allogeneic transplant rejection, cGMP-PKG signaling pathway, TGF-beta signaling pathway, and bile secretion. Results are shown below. Figure 7 .

[0077] Example 5

[0078] Construction of a network diagram of effective components, intersecting genes, and pathways in traditional Chinese medicine compositions for treating breast cancer.

[0079] In order to obtain the main active ingredients and core targets of the traditional Chinese medicine composition for treating breast cancer of the present invention, based on the already obtained effective ingredients and their targets of action of compound drugs, potential targets of compound drugs for treating diseases, and signaling pathways enriched by intersecting genes, a Network and Type table was established and imported into Cytoscape software for visualization to obtain a network diagram of compound effective ingredients-intersecting genes-pathway.

[0080] Based on the obtained effective components of the compound drug, abnormal BC targets, and the top 20 pathways with the lowest FDR values, Network and Type tables were constructed and imported into Cytoscape software for visualization, resulting in a network diagram of compound effective components, intersecting genes, and pathways. Cytoscape software was then used to draw and analyze the relationship network diagram, yielding a total of 496 nodes (including 20 pathways, 158 effective components, and 318 targets) and 2561 relationships. Results are shown below. Figure 10 .

[0081] The network topology parameters between the active ingredients, intersecting genes, and pathways of the compound were analyzed using built-in tools in Cytoscape software. Based on the degree value (node ​​connectivity), the core components and core targets were identified. Cytoscape network analysis showed that quercetin had a degree of 557 and a closeness-centrality of 0.4717, predicting quercetin to be the main component of the compound drug for treating BC, followed by luteolin (degree of 226, closeness-centrality of 0.4239), kaempferol, beta-sitosterol, and artemisinin. The results are shown in Table 2.

[0082] Table 2. Main active ingredients and network node characteristic parameters of traditional Chinese medicine compositions for treating breast cancer.

[0083]

[0084] Example 6

[0085] Traditional Chinese medicine composition for treating breast cancer - target molecule docking

[0086] After performing topology analysis on the network, core compounds and their intersection targets were screened for molecular docking. The 3D structures of the components in the TCMSP database were converted from mol2 format to pdbqt format using Open Babel software. Water molecules and small molecule components of the key targets in the PDB database were removed from their 3D structures using PyMoL software, resulting in pdbqt format files. Hydrogen was then added to the key targets using Autodocktools software. Finally, molecular docking was completed using Autodock vina 1.1.2, and microscopic and macroscopic diagrams of the docking mode were plotted using PyMol software.

[0087] After topological analysis of the network, the core compounds quercetin, luteolin, kaempferol, beta-sitosterol, and artemetin were screened and molecularly docked with the core intersection targets PTGS2, PTGS1, NCOA2, ADRB2, and SCN5A, respectively. (See attached diagram.) Figure 11 AE. The lower the binding energy of molecular docking, the greater the binding potential between the two molecules. The results of molecular docking between the core compound and the core target were mostly less than -7.0 kcal / mol, and the remaining values ​​were also close to -7.0 kcal / mol, indicating that most targets and components had good binding activity and tight binding. The results are shown in Table 3.

[0088] Table 3. Molecular docking binding energy between core compounds and core targets in traditional Chinese medicine compositions for treating breast cancer.

[0089]

[0090]

[0091] Although the present invention has been described in detail in the embodiments through general description, specific implementation and experiment, any modifications or improvements that can be made without departing from the core of the present invention shall fall within the scope of protection of the present invention.

Claims

1. A traditional Chinese medicine composition for treating breast cancer, characterized in that, The traditional Chinese medicine composition contains the following raw materials: Bupleurum, Angelica sinensis, Paeonia lactiflora, Curcuma longa, Taraxacum mongolicum, Echinops latifolius, Scutellaria barbata, Prunella vulgaris, Fritillaria thunbergii, Fritillaria thunbergii, Gleditsia sinensis thorns, Trionyx sinensis, Salvia miltiorrhiza, and Ligustrum lucidum.

2. The traditional Chinese medicine composition for treating breast cancer according to claim 1, characterized in that, The traditional Chinese medicine composition comprises the following raw materials in parts by weight: Bupleurum chinense 10-15 parts, Angelica sinensis 8-12 parts, Paeonia lactiflora 8-12 parts, Curcuma longa 8-12 parts, Taraxacum mongolicum 6-9 parts, Echinops latifolius 6-9 parts, Scutellaria barbata 6-9 parts, Prunella vulgaris 6-9 parts, Fritillaria thunbergii 3-6 parts, Fritillaria thunbergii 10-12 parts, Gleditsia sinensis 6-9 parts, Trionyx sinensis 6-9 parts, Salvia miltiorrhiza 6-12 parts, and Ligustrum lucidum 10-15 parts.

3. The traditional Chinese medicine composition for treating breast cancer according to claim 2, characterized in that, The traditional Chinese medicine composition contains the following raw materials in parts by weight: Bupleurum chinense 15 parts, Angelica sinensis 12 parts, Paeonia lactiflora 12 parts, Curcuma longa 9 parts, Taraxacum mongolicum 9 parts, Echinops latifolius 9 parts, Scutellaria barbata 9 parts, Prunella vulgaris 9 parts, Fritillaria thunbergii 6 parts, Fritillaria thunbergii 12 parts, Gleditsia sinensis thorns 9 parts, Trionyx sinensis 9 parts, Salvia miltiorrhiza 12 parts, and Ligustrum lucidum 15 parts.

4. A traditional Chinese medicine preparation for treating breast cancer, characterized in that, The traditional Chinese medicine preparation is prepared from the traditional Chinese medicine composition for treating breast cancer as described in any one of claims 1-3.

5. The traditional Chinese medicine preparation for treating breast cancer according to claim 4, characterized in that, The traditional Chinese medicine preparation is a decoction, which is prepared according to the following steps: (1) Weigh the raw materials according to the weight parts, add 5-10 times the weight parts of water and soak for 30-120 minutes; (2) Heat and decoct the soaked raw materials together with water, filter, and collect the filtrate; (3) Add 5-10 times the weight of water to the filter residue, boil, filter, and collect the filtrate; (4) Combine the filtrates obtained after two decoctions and heat them to concentrate to obtain the decoction.

6. The traditional Chinese medicine preparation for treating breast cancer according to claim 5, characterized in that, The cooking time for both simmering sessions is 30-60 minutes.

7. The traditional Chinese medicine preparation for treating breast cancer according to claim 5, characterized in that, The filtrates are combined and then concentrated by heating to 10%-20% of their original volume.

8. The use of the traditional Chinese medicine composition for treating breast cancer according to any one of claims 1-3 in the preparation of a medicament for treating breast cancer.

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