A traditional Chinese medicine composition for treating breast nodules, its preparation method and uses

Abstract

The present invention provides a traditional Chinese medicine composition for treating breast nodules, which is prepared from the following raw materials in the following weight ratios: 5-15 parts of vinegar-processed turtle shell, 8-12 parts of bolbostemma paniculatum, 3-6 parts of tangerine leaves, 8-12 parts of green tangerine peel, 5-15 parts of cyperus rotundus, 5-15 parts of rhizoma corydalis, 5-10 parts of myrrh, 8-12 parts of pleione bulbocodioides var. chinensis, 4-10 parts of ground beetle, 5-15 parts of zedoary, 4-10 parts of semiaquilegia adoxoides, 5-15 parts of astragalus membranaceus, 2-10 parts of angelica sinensis, 2-10 parts of cuscuta chinensis, and 2-10 parts of morinda officinalis. The present invention also provides a preparation method and use of the pharmaceutical composition. The drug of the present invention has high safety, and the efficacy and clinical trials prove that it has significant efficacy in treating breast hyperplasia, especially for breast nodules, providing a new option for clinical treatment of breast nodules and breast hyperplasia.

Description

Technical Field

[0001] The present invention relates to a traditional Chinese medicine composition for treating breast nodules and belongs to the field of traditional Chinese medicine. Background Art

[0002] Modern medicine believes that the causes of breast nodules in women are usually due to the imbalance of estrogen and progesterone, endocrine hormone disorders, and sex hormone disorders. It is usually also related to mood, stress, living habits, and living environment to a certain extent.

[0003] Traditional Chinese medicine does not have the disease name of breast nodules. Generally, this disease is considered to belong to "Ru Pi", "Ru Pi", "Ru Su" in traditional Chinese medicine, etc. Physicians in past dynasties have different descriptions and names for breast nodules. This disease belongs to the category of diseases such as "Ru Pi", "Ru Pi", "Ru Zhong Jie He", "Ru Su", "Ru Li" in traditional Chinese medicine. Regarding the etiology and pathogenesis of this disease, physicians in past dynasties have their own opinions. Wang Kentang's "Zheng Zhi Zhun Sheng" believes that "due to worry, anger, depression, and accumulation over time, the liver qi rebels horizontally and thus forms addiction nodules". Article 131 of Treatise on Febrile Diseases "When a disease occurs in yang and is treated with purgation, heat enters and forms chest binding. When it occurs in yin and is treated with purgation, it forms stuffiness...". Stuffiness means pi, and pi is the stagnation of qi and blood circulation. Distending pain appears in any part of the human body, appearing and disappearing from time to time, getting better and worse from time to time, and blood stasis and amenorrhea, which is very consistent with the clinical manifestations of Ru Pi. "Gao's Experience in Surgical Diseases of Traditional Chinese Medicine" "When the liver qi has a stagnation and the qi cannot be released smoothly, the formation of swelling and hardness". Chen Shigong in the Ming Dynasty believed that this disease "is mostly caused by thinking and worrying injuring the spleen and anger injuring the liver, resulting in stagnation and formation". It clarifies that liver qi stagnation, emotional injury, and blood stasis blockage are the main etiology and pathogenesis of this disease. Regarding the understanding of the treatment of this disease, Huangdi Neijing says: All diseases are caused by qi. "Compilation of External Syndrome Medical Records" points out: "When treating breast diseases, if the liver and gallbladder wood qi is stagnant, it will be carbuncle; if the vital qi is deficient, it will be cancer. Phlegm and qi condense to form pi, stuffiness, and nodules. Among various prescriptions, products for regulating qi and dredging collaterals are added to dredge the breast collaterals, and treating the liver will make the swelling disappear by itself", which has a greater impact on later physicians. And Yu Tinghong's view that "the nodules in the breast, although it is said to be a disease, its root cause lies in the kidney" provides a theoretical basis for the treatment method of regulating Chong and Ren meridians and warming the kidney yang for this disease.

[0004] Breast nodules are a symptom, commonly seen in breast hyperplasia and breast tumorous diseases, including benign breast tumors (including breast fibroids, phyllodes tumors, etc.) and malignant breast tumors (breast cancer). Breast nodules can occur in any age group of women. Currently, except for surgical treatment, there is no better drug prevention and treatment. Especially for nodules with irregular shapes detected by B-ultrasound; and breast atypical hyperplasia; breast adenosis, etc. These are neither benign nor have developed to the malignant level, and at this time, it is very necessary to use drugs for treatment to soften and disperse the nodules and control the progression and deterioration of the disease.

[0005] Application No. 201310405052.2, Invention Title: A Traditional Chinese Medicine Granule for Treating Breast Lumps and Its Preparation Method. It discloses a traditional Chinese medicine granule for treating breast lumps and its preparation method, which is mainly prepared from the following raw materials in the following weight ratios: Cyperus rotundus, Cremastra appendiculata, Bolbostemma paniculatum, Crataegus pinnatifida, Manis pentadactyla, Rhaponticum uniflorum, Curcuma aromatica, Rhizoma zedoariae, Gleditsia sinensis spine, Trionyx sinensis carapace, Trichosanthes kirilowii, Allium macrostemon, American ginseng, Vaccaria segetalis, Carthamus tinctorius. The traditional Chinese medicine granule of this invention can effectively relieve qi stagnation caused by breast lumps, promote blood circulation and relieve pain, and has a good therapeutic effect on breast lumps. Application No. 201210219219.1, Invention Title: A Traditional Chinese Medicine Prescription for Treating Mammary Lobular Hyperplasia and Fibroadenoma of Breast. It discloses a traditional Chinese medicine prescription for treating mammary lobular hyperplasia and fibroadenoma of breast, and its composition is as follows: Paeonia lactiflora 6 - 10 g, Angelica sinensis 6 - 12 g, Poria cocos 6 - 10 g, Atractylodes macrocephala 6 - 12 g, Bupleurum chinense 3 - 6 g, Cyperus rotundus 6 - 10 g, Curcuma aromatica 6 - 9 g, Citrus reticulata Blanco var. austera 6 - 12 g, Corydalis yanhusuo 6 - 9 g, Sparganium stoloniferum 6 - 9 g, Rhizoma zedoariae 6 - 9 g, Salvia miltiorrhiza 6 - 10 g, Carthamus tinctorius 3 - 6 g, Crataegus pinnatifida 6 - 10 g, Vaccaria segetalis 6 - 10 g, Piper nodorum 9 - 12 g, Trionyx sinensis carapace 10 - 15 g, Ostrea gigas thunbergii 10 - 20 g, Bolbostemma paniculatum 6 - 9 g, Trichosanthes kirilowii pericarpium 6 - 9 g, Sinapis alba 6 - 12 g, Trichosanthes kirilowii Maxim 6 - 10 g, Glycyrrhiza uralensis 5 g. Due to the different etiologies and pathogenesis of "breast lumps", the dialectics also has differences. The currently disclosed prescriptions cannot meet the clinical needs. Summary of the Invention

[0006] The technical solution of the present invention provides a traditional Chinese medicine composition for treating breast nodules, its preparation method and uses.

[0007] The present invention provides a traditional Chinese medicine composition for treating breast nodules, which is prepared from the following raw materials in the following weight ratios:

[0008] Vinegar-processed Trionyx sinensis carapace 5 - 15 parts, Bolbostemma paniculatum 8 - 12 parts, Citrus reticulata Blanco var. austera 3 - 6 parts, Citrus reticulata Blanco var. austera 8 - 12 parts, Cyperus rotundus 5 - 15 parts, Corydalis yanhusuo 5 - 15 parts, Myrrha 5 - 10 parts, Cremastra appendiculata 8 - 12 parts, Eupolyphaga sinensis 4 - 10 parts, Rhizoma zedoariae 5 - 15 parts, Semen euphorbiae helioscopiae 4 - 10 parts, Astragalus membranaceus 5 - 15 parts, Angelica sinensis 2 - 10 parts, Cuscuta chinensis 2 - 10 parts, Morinda officinalis 2 - 10 parts.

[0009] Further preferably, it is prepared from the following raw materials in the following weight ratios: Vinegar-processed Trionyx sinensis carapace 8 - 12 parts, Bolbostemma paniculatum 8 - 12 parts, Citrus reticulata Blanco var. austera 4 - 6 parts, Citrus reticulata Blanco var. austera 4.8 - 7.2 parts, Cyperus rotundus 6.4 - 9.6 parts, Corydalis yanhusuo 6.4 - 9.6 parts, Myrrha 4 - 6 parts, Cremastra appendiculata 4.8 - 7.2 parts, Eupolyphaga sinensis 4 - 6 parts, Rhizoma zedoariae 4.8 - 7.2 parts, Semen euphorbiae helioscopiae 4 - 6 parts, Astragalus membranaceus 6.4 - 9.6 parts, Angelica sinensis 4.8 - 7.2 parts, Cuscuta chinensis 4.8 - 7.2 parts, Morinda officinalis 4.8 - 7.2 parts.

[0010] Further preferably, it is prepared from the raw medicinal materials with the following weight ratios:

[0011] 10 parts of vinegar-processed turtle shell, 10 parts of bolbostemma rhizome, 5 parts of tangerine leaf, 6 parts of green tangerine peel, 8 parts of cyperus rotundus, 8 parts of rhizoma corydalis, 5 parts of myrrh, 6 parts of pseudobulbus pleiones, 5 parts of ground beetle, 6 parts of zedoary, 5 parts of semiaquilegia root tuber, 8 parts of astragalus membranaceus, 6 parts of angelica sinensis, 6 parts of cuscuta chinensis, 6 parts of morinda officinalis.

[0012] The traditional Chinese medicine composition of the present invention takes the crude drug powder or water or organic solvent extract of the traditional Chinese medicine composition as the active ingredient, and is prepared into a pharmaceutically common preparation by adding pharmaceutically acceptable excipients or auxiliary components.

[0013] Among them, the preparation is an oral preparation.

[0014] Among them, the oral preparation is tablets, capsules, pills, granules, oral liquids.

[0015] The present invention also provides a method for preparing the traditional Chinese medicine composition, which includes the following steps:

[0016] a. Weigh the raw medicinal materials with each weight ratio;

[0017] b. Add 60%-90% ethanol to bolbostemma rhizome, tangerine leaf, green tangerine peel, cyperus rotundus, rhizoma corydalis, myrrh, pseudobulbus pleiones, zedoary, and reflux extract for 2-4 times, and recover the ethanol from the ethanol extract;

[0018] c. Mix the medicinal residues after the above ethanol extraction with semiaquilegia root tuber, astragalus membranaceus, angelica sinensis, cuscuta chinensis, morinda officinalis, decoct with water, combine the water extracts, discard the medicinal residues, combine the decocted liquid with the concentrated liquid after recovering ethanol in step b, concentrate and dry;

[0019] d. Powder vinegar-processed turtle shell and ground beetle, mix with the dried product in step c, and then add pharmaceutically acceptable excipients or auxiliary components to prepare a pharmaceutically common preparation.

[0020] The present invention also provides the use of the traditional Chinese medicine composition in the preparation of a drug for treating breast nodules.

[0021] The present invention also provides the use of the traditional Chinese medicine composition in the preparation of a drug for treating breast nodules or breast hyperplasia.

[0022] The present invention also provides the use of the traditional Chinese medicine composition in the preparation of a drug with the functions of soothing the liver and regulating qi, promoting blood circulation to remove blood stasis, regulating thoroughfare and conception vessels, and warming and tonifying kidney yang.

[0023] The pharmaceutical composition of the present invention uses stir-fried turtle shell and bolbostemma paniculatum as the monarch drugs. Stir-fried turtle shell is a "precious and nourishing" herb in traditional Chinese medicine, a rare and precious traditional Chinese medicine. It tastes salty, enters the liver meridian, is good at softening hardness and dissipating nodules, and is effective in treating masses and accumulations. Bolbostemma paniculatum is slightly cold and is good at dissipating nodules and relieving swelling. Modern medical research shows that bolbostemma paniculatum has the effect of treating nodules and tumors. Bolbostemma paniculatum is non-toxic and inexpensive. The ministerial drugs are orange leaves, green tangerine peel, cyperus rotundus, rhizoma corydalis, and myrrh. Orange leaves are of neutral nature and enter the liver meridian. According to research, the flavonoid compounds in orange leaves have anti-tumor effects. Orange leaves and green tangerine peel can enhance the effect of soothing the liver and regulating qi, while cyperus rotundus is pungent and good at relieving liver depression, and rhizoma corydalis and myrrh are pungent and warm, having the effect of promoting blood circulation to relieve pain. The adjuvant drugs are pleione bulbocodioides, ground beetle, curcuma zedoaria, and semiaquilegia adoxoides. Pleione bulbocodioides tastes sweet and pungent, enters the liver meridian, and is good at clearing heat and detoxifying, resolving phlegm and dissipating nodules. Ground beetle tastes salty and enters the liver meridian, promoting blood circulation to disperse stasis and dredging collaterals. Curcuma zedoaria is pungent, dispersing, warm and dredging, enters the liver meridian, promoting blood circulation to remove blood stasis, regulating qi to relieve pain, and eliminating masses. Semiaquilegia adoxoides enhances the effect of dissipating nodules. The envoy drugs are astragalus membranaceus, angelica sinensis, cuscutae chinensis, and morinda officinalis. Astragalus membranaceus and angelica sinensis strengthen healthy qi, fearing that promoting blood circulation to remove blood stasis may be excessive. Then, cuscutae chinensis and morinda officinalis regulate the thoroughfare and conception vessels and warm the kidney yang.

[0024] The diseases treated by the drugs of the present invention are stubborn diseases. Wang Qingren, a famous doctor in the Qing Dynasty, advocated "treating diseases quickly" and "removing blood stasis and promoting blood circulation". This formula uses stir-fried turtle shell and pleione bulbocodioides, which are good at softening hardness and dissipating nodules, as the monarch drugs, utilizes the effect of promoting blood circulation to remove blood stasis of ground beetle, curcuma zedoaria, rhizoma corydalis, and myrrh, and then timely uses orange leaves, cyperus rotundus, etc. to soothe the liver qi, showing the combined effect of softening hardness, soothing the liver, promoting blood circulation, and dissipating nodules. At the same time, it can also improve the immune function of patients.

[0025] The drugs of the present invention are highly safe. In terms of single-dose toxicity: For normal mice, when administered at the maximum single-dose (580.8 g / kg), which is 1584 times the clinical adult daily dose (0.3666 g / kg), no obvious acute toxicity occurred. In terms of repeated-dose toxicity: Female SD rats were continuously administered at a dose of 44 g / kg (equivalent to 120 times the clinical adult daily dose) for 13 weeks, and no abnormalities were found in various indicators such as histopathology, biochemistry, and hematology of the rats. Pharmacodynamic and clinical trials have proved that the drugs of the present invention are effective in treating mammary gland hyperplasia, especially for mammary nodules, providing a new option for the clinical treatment of mammary nodules and mammary gland hyperplasia. Description of the Drawings

[0026] Figure 1Improvement of the morphological changes of mammary glands in rats with mammary gland hyperplasia by the pharmaceutical composition (A. Blank group: few acini in the mammary lobules were observed under microscopy, and there was no secretion in the acini and ducts; B. Model group: the volume of the mammary lobules increased under microscopy, the number of acini in the lobules increased, the ducts dilated, and there was a large amount of secretion in the lumen; C. Tamoxifen (3 mg / kg) group, D-F. Pharmaceutical composition API 2.26 g / kg, 4.52 g / kg, 9.02 g / kg groups: compared with the model group, the number of acini in the mammary lobules decreased under microscopy, and the number of dilated ducts decreased. (Bar = 300 μm, ×100, the hyperplastic mammary lobules or dilated ducts are indicated by the black arrows)). Specific implementation method

[0027] Example 1 Preparation of the drug of the present invention

[0028] Weigh the API: 100 g of stir-fried turtle shell with vinegar, 100 g of bolbostemma paniculatum, 50 g of citrus leaf, 60 g of green tangerine peel, 80 g of cyperus rotundus, 80 g of Corydalis yanhusuo, 50 g of myrrh, 60 g of pseudobulbus cremastrae seu pleiones, 50 g of ground beetle, 60 g of zedoary, 50 g of semiaquilegia adoxoides, 80 g of astragalus membranaceus, 60 g of angelica sinensis, 60 g of cuscuta chinensis, 60 g of morinda officinalis

[0029] Take bolbostemma paniculatum, citrus leaf, green tangerine peel, cyperus rotundus, Corydalis yanhusuo, myrrh, pseudobulbus cremastrae seu pleiones, zedoary, add 6 times the amount of 75% ethanol, reflux and extract 2-4 times, 1 hour each time, and recover the ethanol from the ethanol extract

[0030] The residue after the above ethanol extraction is mixed with semiaquilegia adoxoides, astragalus membranaceus, angelica sinensis, cuscuta chinensis, morinda officinalis in the prescription, add 6 times the amount of water, decoct 2-4 times, 2 hours each time, filter, combine the water extracts, discard the residue, combine the decoction with the concentrated solution after recovering the ethanol above, under -0.015 - 0.030 MPa, concentrate under reduced pressure at a temperature of 80 °C until the relative density of the extract is 1.05 - 1.15, spray-dry at 70 °C to obtain dry extract powder, then mix it with the sterilized fine powder of stir-fried turtle shell with vinegar and ground beetle reserved above to obtain the API powder, fill it into capsules to prepare capsule preparations

[0031] Example 2 Preparation of the tablets of the drug of the present invention

[0032] Weigh the API: 12 g of stir-fried turtle shell with vinegar, 12 g of bolbostemma paniculatum, 6 g of citrus leaf, 7.2 g of green tangerine peel, 9.6 g of cyperus rotundus, 9.6 g of Corydalis yanhusuo, 6 g of myrrh, 7.2 g of pseudobulbus cremastrae seu pleiones, 6 g of ground beetle, 7.2 g of zedoary, 6 g of semiaquilegia adoxoides, 9.6 g of astragalus membranaceus, 7.2 g of angelica sinensis, 7.2 g of cuscuta chinensis, 7.2 g of morinda officinalis, prepare according to the method of Example 1, and directly compress the mixed powder into tablets

[0033] Example 3 Preparation of the capsule preparations of the drug of the present invention

[0034] 8g of vinegar turtle shell, 8g of ground Fritillaria, 4g of citrus leaf, 4.8g of green tangerine peel, 6.4g of Cyperus rotundus, 6.4g of Corydalis yanhusuo, 4g of Myrrh, 4.8g of Tripterygium wilfordii, 4g of Eupolyphaga sinensis, 4.8g of Curcuma zedoaria, 4g of Aquilegia vulgaris, 6.4g of Astragalus membranaceus, 4.8g of Angelica sinensis, 4.8g of Cuscuta australis, and 4.8g of Morinda officinalis were prepared into capsules according to the method of Example 1.

[0035] The beneficial effects of the present invention are demonstrated by specific pharmacodynamic tests or clinical trials as follows.

[0036] Test Example 1 The therapeutic effect of the drug of the present invention on a rat model of mammary hyperplasia

[0037] 1. Preparation of experimental drugs and reagents

[0038] Preparation of modeling drugs: Estradiol benzoate injection (purchased from Ningbo Second Hormone Factory, production batch number: 171026, specification: 4mg / 2ml) was diluted to 0.5mg / ml with injection soybean oil (purchased from Zhejiang Tianyushan Medicinal Oil Co., Ltd., batch number: 171212, specification: 1000ml) before use, and progesterone injection (purchased from Tianjin Jinyao Pharmaceutical Co., Ltd., batch number: 171028, specification: 2mg / ml) was diluted to 5mg / ml with injection soybean oil before use. The injection volume was 1ml / kg, that is, the dosage of estradiol benzoate 0.5mg / (kg·d) and progesterone 5mg / (kg·d) were administered to each rat.

[0039] Positive control drug: Tamoxifen citrate tablets, Yangtze River Pharmaceutical Group Co., Ltd., specification (based on tamoxifen): 10 mg / tablet, production batch number: 18030211, approval number: National Medicine Standard H32021472. Grind into powder before use, prepare into a drug solution with a concentration of 0.38 mg / ml with distilled water, and the dosage volume for each rat is 8 ml / (kg·d), that is, the dosage for each rat is 3 mg / (kg·d).

[0040] Estradiol (E2), progesterone (P), and prolactin (PRL) ELISA enzyme-linked immunosorbent assay kits (Shanghai Youxuan Biotechnology Co., Ltd., batch number: 201809); other reagents were all domestic analytical grade.

[0041] 2 Experimental animals and instruments

[0042] Female clean-grade SD rats, 155-165 g, non-pregnant, were purchased from the Experimental Animal Research Institute of Chongqing Institute of Traditional Chinese Medicine, license number SCXK (渝) 2017-0003. The experimental animal quality certificate number is 0001542. They were kept in the Experimental Animal Center of the School of Pharmacy of Southwest University, with the experimental animal use license number SYXK (渝) 2014-0002. The temperature was 20-25 °C, the relative humidity was 40%-60%, and the light-dark cycle was 12h / 12h.

[0043] MNT-150 vernier caliper (Shanghai Menite Industrial Co., Ltd.), EL204 electronic balance (Mettler Toledo Instrument Co., Ltd.), R134A low-temperature high-speed centrifuge (Eppendorf), ELX800 microplate reader (Gene Co., Ltd.), CM1900 paraffin slicer (Leica Microsystems Trading Co., Ltd.), and DM4000 electron microscope (Leica Instrument Co., Ltd., Germany).

[0044] 3 Experimental design

[0045] 3.1 Reproduction of the rat mammary hyperplasia model

[0046] Model replication method: 66 female non-pregnant clean-grade SD rats were taken, adaptively raised for one week, and randomly divided into a blank group (n=10) and a modeling group (n=56), and the model was established according to the method determined in the preliminary experiment. The specific operation is as follows: the rats in the modeling group were alternately injected with 0.5 mg / kg of estradiol benzoate in the inner muscles of the hind limbs once a day for 25 consecutive days; then progesterone 5 mg / kg was injected intramuscularly once a day for 5 consecutive days for a total of 30 days. From the 31st day, 0.5 mg / kg of estradiol benzoate was injected intramuscularly every 3 days. The blank group was intramuscularly injected with an equal volume of soybean oil (the diluent for estradiol benzoate and progesterone injection).

[0047] Model success criteria: 30 days after modeling, 6 rats were randomly selected from the modeling group. The height and diameter of the second pair of nipples in the modeling group were significantly higher than those in the blank group (P<0.01). Pathological sections were observed to show corresponding pathological changes in the glandular ducts, alveoli, and glandular lobules of the nipples in the modeling group, and the modeling was considered successful.

[0048] 3.2 Setting of drug dosage

[0049] The sample used in this implementation is the raw material powder of the Chinese medicine composition, and each 1g of powder is equivalent to 6.6g of crude drug. The recommended daily clinical dosage of this Chinese medicine composition is 22g / 60kg of crude drug. Take 1 times, 2 times, and 4 times the clinical dosage as the test dose, convert it to rats, and set the crude drug content of 2.26g / kg as the low-dose group, the crude drug content of 4.52g / kg as the medium-dose group, and the crude drug content of 9.04g / kg as the high-dose group.

[0050] 3.3 Grouping and Dosing

[0051] Grouping and inter-group balance test: 30 days after modeling, the rats meeting the successful modeling criteria were randomly divided into 5 groups according to the random number table, namely the model group, the tamoxifen group, and the low, medium, and high-dose groups of the crude drug powder of the traditional Chinese medicine composition, with 10 rats in each group. Statistical processing was performed on the mean values of nipple height and diameter among the 5 groups of animals after modeling. Only when P>0.05 indicates no significant difference can they be included in the next experiment.

[0052] Drug administration dose and cycle: Each group was given the corresponding drug according to the following table. The administration volume was 0.8 ml / 100 g. The blank group and the model group were gavaged with an equal volume of distilled water. It was administered once a day for 90 days. (See Table 1)

[0053] Table 1 Experimental grouping and doses of the therapeutic effect of the drug composition on rat mammary gland hyperplasia

[0054]

[0055] 3.4 Measurement of breast diameter and nipple height

[0056] The height of the second pair of nipples and the breast diameter of the rats were measured before modeling, before and after drug administration using a precision vernier caliper.

[0057] 3.5 Determination of organ coefficients

[0058] The rats were sacrificed 24 hours after the last drug administration. The body weight, thymus, spleen, uterus, and ovary weights were measured, and the organ coefficients were calculated according to the following formula. Organ coefficient = organ mass (g) / body weight (g) × 100%.

[0059] 3.6 Detection of serum hormone levels

[0060] 24 hours after the last drug administration, the rats were anesthetized with 3.5% chloral hydrate, and blood was taken from the abdominal aorta. After centrifugation at 3000 r / min for 15 minutes at 4°C, the serum was separated and stored frozen at -80°C. According to the kit instructions, the contents of estradiol (E2), progesterone (P), and prolactin (PRL) were measured by enzyme-linked immunosorbent assay.

[0061] 3.7 Histopathological examination of mammary gland tissue

[0062] After blood collection, the second pair of intact breasts of the rats were aseptically removed and fixed with 4% paraformaldehyde solution. After overnight at 4°C, they were dehydrated step by step with ethanol, embedded in paraffin, sectioned, stained with HE, and the hyperplasia of rat mammary gland tissue was observed under a light microscope. The specific operations are as follows (see Table 2).

[0063] Table 2 Experimental steps of HE staining of paraffin sections

[0064]

[0065]

[0066] After the coverslip is completed, microscopic examination is carried out, and image acquisition and analysis are performed. 3.8 Detection of ER and PR expression in breast tissue

[0067] The specific operation is as follows (see Table 3).

[0068] Table 3 Immunohistochemistry experimental steps for paraffin sections

[0069]

[0070]

[0071] Result judgment: The presence of brown-yellow granules in the cytoplasm and / or nucleus is regarded as positive cells. Five high-power microscopic fields are selected for each specimen for statistics. The integrated optical density value (IOD) and effective area (area) of the brown-yellow area are measured using Image-Pro Plus, and the average optical density (IOD / area) is calculated.

[0072] 3.9 Statistical analysis

[0073] SPSS 22.0 statistical software is used for analysis, and the experimental data are expressed as One-way ANOVA is used for comparison between groups, and t-test is used for significance analysis. P < 0.05 indicates statistically significant differences.

[0074] 4 Experimental results

[0075] 4.1 The pharmaceutical composition significantly reduces the breast diameter and nipple height of rats with mammary gland hyperplasia

[0076] In the blank group, the nipple position of the rats was invisible through the hair. After separating the hair, the nipples were visible as small granular, sticking to the skin, soft, and slightly elevated in some cases. Compared with the blank group, the breast diameter and nipple height of the rats in the model group were significantly increased (P < 0.01), and the nipples were red and swollen or enlarged. The degree of nipple swelling in each drug administration group was alleviated, and the breast diameter and nipple height were smaller than those in the model group (P < 0.01), as shown in Table 4.

[0077] 4.2 The pharmaceutical composition significantly increases the thymus index and decreases the uterine index of rats with mammary gland hyperplasia

[0078] Compared with the blank group, the thymus index of the model group was extremely significantly decreased (P<0.01), and the uterine index was extremely significantly increased (P<0.01), indicating that the immunity of rats decreased, the thymus atrophied, and uterine cysts occurred under the stimulation of sex hormones. Compared with the model group, the thymus index of each dose group of tamoxifen and the drug composition raw material was significantly increased (P<0.05), and the uterine index was extremely significantly decreased (P<0.01), as shown in Table 5. The results showed that the drug composition raw material had the effects of improving uterine cysts and enhancing immunity.

[0079] Table 4 The drug composition significantly reduced the breast diameter and nipple height of rats with mammary hyperplasia

[0080]

[0081] Note: Compared with the blank group, 1) P<0.01; compared with the model group, 2) P<0.01

[0082] Table 5 The drug composition significantly increased the thymus index and decreased the uterine index of rats with mammary hyperplasia

[0083]

[0084] Note: Compared with the blank group, 1) P<0.01; compared with the model group, 2) P<0.05, 3) P<0.01

[0085] 4.3 The drug composition has a significant regulatory effect on the serum hormone content of rats with mammary hyperplasia

[0086] Compared with the blank group, the serum E2 and PRL contents of the model group rats were significantly increased (P<0.01), and the P content was significantly decreased (P<0.01). The low, medium, and high dose groups of tamoxifen and the drug composition raw material could all reduce the serum E2 and PRL contents of rats to varying degrees, and could significantly increase the serum P content of the model group rats (P<0.01). See Table 6.

[0087] Table 6 The drug composition has a significant regulatory effect on the serum hormone content of rats with mammary hyperplasia

[0088]

[0089] Note: Compared with the blank group, 1) P<0.01; compared with the model group, 2) P<0.01

[0090] 4.4 The drug composition significantly reduces the expression of ER and PR in the mammary gland tissue of rats with mammary gland hyperplasia

[0091] Compared with the blank control group, the expression of ER and PR in the mammary gland tissue of rats in the model group was significantly enhanced (P < 0.01). The expression of ER and PR in the mammary gland tissue of rats in the drug administration group decreased, showing a significant difference compared with the model group (P < 0.01). See Table 7.

[0092] Table 7 The drug composition significantly reduces the expression of ER and PR in the mammary gland tissue of rats with mammary gland hyperplasia

[0093]

[0094] Note: Compared with the blank group, 1) P < 0.01; compared with the model group, 2) P < 0.01

[0095] 4.5 The drug composition has a significant improvement effect on the morphology of the mammary gland tissue of rats with mammary gland hyperplasia

[0096] The HE results showed that the number of acini in the mammary gland lobules of rats in the blank group was small, and there was no secretion in the acini and ducts; the volume of the mammary gland lobules of rats in the model group increased, the number of acini in the lobules increased, the ducts dilated, and there was a large amount of secretion in the lumen; compared with the model group, the number of acini in the mammary gland lobules of rats in the tamoxifen group decreased significantly, and the number of dilated ducts and secretion decreased; in the high, medium, and low dose groups of the drug composition raw material, the number of acini in the mammary gland lobules of rats decreased, and the duct diameter decreased significantly. See Figure 1 .

[0097] Test Example 2 Clinical observation of 100 patients with mammary gland nodules treated with the drug of the present invention

[0098] 1 Enrollment of clinical patients

[0099] 100 outpatients were observed. 90% of the patients had received various traditional Chinese and Western medicines in large hospitals, but their conditions did not improve significantly, and some even worsened. All 100 patients were female. The age of the patients was 13 - 74 years old, with an average age of 35 years.

[0100] Age 13 - 25 years old 26 - 35 years old 36 - 45 years old 46 - 52 years old 74 years old Quantity 8 cases 16 cases 46 cases 25 cases 5 cases

[0101] 2 Diagnostic criteria and clinical classification

[0102] Diagnostic Criteria and Clinical Classification: According to the National Traditional Chinese Medicine Clinical Manifestations in 1996, the main manifestations are breast stabbing pain or distending pain accompanied by masses. Some patients are also accompanied by irregular menstruation and mood swings, and some have nipple discharge. There are 5 cases with a family history of breast cancer. Some patients' pain is aggravated before menstruation, and some is aggravated after getting angry. Clinically, single or multiple irregular masses and nodules of varying sizes can be palpated in the breast. The texture is tough, and the boundary is clear, with a regular shape in 72 cases. The boundary is unclear, the shape is irregular, the texture of the mass is tough and hard, and the mobility is poor in 28 cases. All cases have undergone color Doppler ultrasound examination, molybdenum target examination or cytological examination. Except for 1 case of recurrence more than 3 years after breast cancer surgery, other diseases are excluded. The diagnosis is breast nodular disease.

[0103] 3 Treatment Methods

[0104] Treatment Method: The above patients only received the drug of the present invention (capsules prepared in Example 1) for treatment for 20 consecutive days, stopped taking during menstruation, and stopped taking other drugs during the medication period.

[0105] Efficacy Criteria: Refer to the relevant content in the "Guiding Principles for Clinical Research of New Chinese Medicines" and the "Diagnostic Criteria for Hyperplasia of Mammary Glands" for formulation:

[0106] Clinical Cure: The mass and breast pain disappear. The B-ultrasound diagnosis classification is: Class 1, and there is no recurrence within half a year to 3 years after stopping the drug.

[0107] Marked Effect: The maximum diameter of the mass is reduced by one-half, the pain disappears, and the B-ultrasound diagnosis classification changes from Class 4 to Class 3 or Class 2.

[0108] Effective: The maximum diameter of the mass is reduced by one-third, the breast pain disappears, and the B-ultrasound diagnosis changes from Class 3 to Class 2.

[0109] Ineffective: The pain is relieved, the mass shows no obvious softening or reduction, and the B-ultrasound diagnosis remains unchanged.

[0110] 4 Treatment Results

[0111] After 20 days of treatment with the drug of the present invention, among 100 subjects, 85 cases were clinically cured, 7 cases showed marked effect, 6 cases were effective, and 2 cases were ineffective. The clinical treatment result of the drug of the present invention is a cure rate of 85% and a total effective rate of 92.3%.

[0112] Clinical Typical Case 1:

[0113] 1. Name: Yu xx, Gender: Female, Age: 48 years old, Occupation: Farmer, Marital Status: Married.

[0114] Date of Visit: February 16, 2018

[0115] She had a breast-conserving surgery for right breast cancer in a hospital in Luzhou more than 3 years ago. When she had a physical examination on December 5, 2017, it was found that the cancer had recurred. The hospital asked the patient to be hospitalized for surgery and chemotherapy again. Due to the patient's fear of surgery and chemotherapy, she came to the outpatient department for treatment on February 16, 2018. The B-ultrasound showed: In the glandular layer at the 11 o'clock position of the right breast, a hypoechoic mass with a size of about 0.73*0.49 cm was detected. The shape was irregular, the boundary was unclear, and there were patchy slightly hyperechoic areas inside. CDFl: A little star-like blood flow signal was seen around the mass. In the glandular layer at the 6 o'clock position of the right breast gland, a mass with a size of about 0.86*0.46 cm was detected, with an irregular shape and a clear boundary. Multiple hypoechoic areas were detected in the right axilla, the largest being about 1.15*0.48 cm

[0116] Ultrasound tip: Solid mass in the right breast, Bl-RADS-3-4 category

[0117] The patient was observed to have a sallow and emaciated face, a worried look, a red tongue with a greasy coating, and a taut and stringy pulse

[0118] Palpation: A mass with a size of about 0.9*0.7 cm was detected at the 11 o'clock position of the right breast. It was hard, not movable, and the boundary was unclear

[0119] Diagnosis: Breast cancer (recurrence after breast cancer surgery)

[0120] Treatment: The patient was given the drug of the present invention for 4 courses of treatment (80 days), 3 times a day

[0121] The patient was instructed to avoid eating pungent and spicy foods. During the medication period, other medications were discontinued

[0122] On July 16, 2018, B-ultrasound examination was performed in the Affiliated Hospital of Southwest Medical University

[0123] Diagnostic opinion: After breast-conserving surgery for the right breast; US-Bl-RADS 1 category for both breasts

[0124] Follow-up for more than 4 years showed no recurrence

[0125] Clinical typical case 2:

[0126] Name: Yang xx, Gender: Female, Age: 37 years old, Occupation: Business, Marital status: Married

[0127] Date of visit: November 23, 2019

[0128] The patient has felt bilateral breast distending pain for over 2 months, which has worsened in the past week. After an ultrasound examination at West China Hospital of Sichuan University, it is indicated that: a cystic-solid mixed echo nodule with a size of approximately 13*6*7 is detected at the 10 o'clock position of the right breast. The boundary is relatively clear, the shape is irregular, there are multiple septations inside, and multiple punctate and patchy strong echoes can be seen around. No obvious blood flow signal is detected. Additionally, anechoic nodules with sizes of 2-5 mm and 3-9 mm are detected in the left and right breasts respectively. Some have poor sound transmission, and patchy strong echoes can be seen on the cyst walls of some. No blood flow signal is detected.

[0129] Diagnostic hint:

[0130] Cystic-solid nodule with calcification at the 10 o'clock position of the right breast. (Bl—RADS category 4A)

[0131] Bilateral breast cysts, with some cystic fluid being viscous. (Bl—RADS category 3)

[0132] Palpation: A mass is felt at the 10 o'clock direction on the right side, with a size of approximately 1.4 cm 3 or so. The surface is not smooth, it is firm and tough in texture, and it is not very mobile.

[0133] Diagnosis: Breast nodulation (bilateral breast nodules), more severe on the right side.

[0134] Upon observing the patient, the face is pale and without luster, the eyes look worried, the tongue is red with yellow coating, and the pulse is string-like and weak.

[0135] Treatment: Administer the drug of the present invention for 3 courses of treatment (60 days), 3 times a day. During the medication period, other drugs should be discontinued. Spicy and fishy foods should be avoided.

[0136] In 2020, an ultrasound examination was conducted at the First People's Hospital of Chengdu: bilateral breasts

[0137] (Bl—RADS category 2)

[0138] After 3 years of follow-up, no recurrence was found.

Claims

1. A traditional Chinese medicine composition for treating breast nodules, characterized in that: It is prepared from the following raw medicinal materials in the following weight ratios: 10 parts of vinegar-processed turtle shell, 10 parts of bolbostemma rhizome, 5 parts of tangerine leaf, 6 parts of green tangerine peel, 8 parts of cyperus rotundus, 8 parts of Corydalis yanhusuo, 5 parts of myrrh, 6 parts of pseudobulbus pleiones, 5 parts of ground beetle, 6 parts of zedoary, 5 parts of Semen euphorbiae helioscopiae, 8 parts of astragalus membranaceus, 6 parts of angelica sinensis, 6 parts of cuscuta chinensis, 6 parts of morinda officinalis.

2. The traditional Chinese medicine composition for treating breast nodules according to claim 1, wherein: It uses the crude drug powder or water or organic solvent extract of the above-mentioned traditional Chinese medicine composition as the active ingredient, and adds pharmaceutically acceptable excipients or auxiliary components to prepare a pharmaceutically common preparation.

3. The traditional Chinese medicine composition for treating breast nodules according to claim 2, wherein: The preparation mentioned above is an oral preparation.

4. The traditional Chinese medicine composition for treating breast nodules according to claim 3, characterized in that: The oral preparation mentioned above is tablets, capsules, pills, granules, or oral liquids.

5. A method for preparing the traditional Chinese medicine composition according to any one of claims 1-4, characterized in that: It includes the following steps: a. Weigh the raw medicinal materials in the respective weight ratios; b. Add 60%-90% ethanol to bolbostemma rhizome, tangerine leaf, green tangerine peel, cyperus rotundus, Corydalis yanhusuo, myrrh, pseudobulbus pleiones, and zedoary, and reflux and extract 2-4 times. Recover the ethanol from the ethanol extract; c. Mix the medicinal residues after the above ethanol extraction with Semen euphorbiae helioscopiae, astragalus membranaceus, angelica sinensis, cuscuta chinensis, and morinda officinalis, decoct with water, combine the water extracts, discard the medicinal residues, combine the decoction with the concentrated solution after recovering ethanol in step b, concentrate, and dry; d. Powder vinegar-processed turtle shell and ground beetle, mix with the dried product in step c, and then add pharmaceutically acceptable excipients or auxiliary components to prepare a pharmaceutically common preparation.

6. Use of the traditional Chinese medicine composition according to any one of claims 1-4 in the preparation of a drug for treating breast nodules.

7. Use of the traditional Chinese medicine composition according to any one of claims 1-4 in the preparation of a drug for treating breast masses or breast hyperplasia.

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