A traditional Chinese medicine monomer composition and use thereof for preparing a drug for treating breast cancer

Abstract

The application discloses a traditional Chinese medicine monomer composition and application of the traditional Chinese medicine monomer composition in preparation of a medicine for treating breast cancer. It is known to those skilled in the art that genistein and alpha-solanine have anti-tumor activity alone, and the inventors of the application accidentally find that the two components have a synergistic effect when used in combination for resisting breast cancer in the research on the anti-tumor mechanism of the two components, which promotes the proposal of the application. The research results of the application reveal that the synergistic effect makes genistein and alpha-solanine have an application prospect of being developed into a medicine for treating breast cancer in the form of a composition.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine and relates to the composition of Chinese medicinal monomers and their applications, specifically to a composition of Chinese medicinal monomers and its use in preparing a drug for treating breast cancer. Background Technology

[0002] Breast cancer is a condition in which mammary epithelial cells proliferate uncontrollably under the influence of various carcinogenic factors. Early symptoms often include breast lumps, nipple discharge, and swollen axillary lymph nodes. In later stages, distant metastasis of cancer cells can lead to multi-organ complications, directly threatening the patient's life. According to the latest global cancer burden data released by the International Agency for Research on Cancer (IARC) of the World Health Organization in 2020, the number of new breast cancer cases has rapidly increased to 2.26 million, officially surpassing lung cancer (2.2 million) for the first time to become the leading cause of cancer death worldwide, accounting for 11.7% of all new cancer cases. In 2020, there were 9.23 million new cancer cases in women globally, accounting for 48% of the total. Among them, 2.26 million were new cases of breast cancer, far exceeding other types of female cancer (colorectal cancer 870,000, lung cancer 770,000, cervical cancer 600,000, thyroid cancer 450,000, endometrial cancer 420,000, stomach cancer 370,000, ovarian cancer 310,000). The incidence rate of breast cancer is equivalent to the combined incidence of the 2nd to 4th ranked cancers, and approximately twice the combined incidence of gynecological cancers such as cervical cancer, ovarian cancer, and endometrial cancer. Of the 4.43 million cancer deaths among women, breast cancer remains the leading cause of death with 680,000 cases, making it the number one killer threatening women's lives.

[0003] The causes of breast cancer are not fully understood. Scientists have not yet found the exact cause of breast cancer, but they have identified many high-risk factors associated with its development. As these high-risk factors accumulate, the risk of developing the disease increases. Current treatment methods for breast cancer mainly include surgery, radiation therapy, endocrine therapy, chemotherapy, molecular targeted therapy, traditional Chinese medicine, and immunotherapy. Most patients undergo a combination of surgery and medication. Traditional Chinese medicine, as the essence of Chinese traditional culture, has contributed many compound prescriptions and preparations. Paclitaxel, a particularly representative drug, is widely used in the clinical treatment of breast cancer patients. Continuing to develop novel anti-breast cancer drugs based on natural products remains a hot topic in new drug development.

[0004] Gentian isoflavone (4',5,7-Trihydroxyisoflavone, C 15 H 10O5 is a polyphenolic compound discovered from the legume *Dendrobium nobile* (Gynostemma pentaphyllum). Its molecular structure is similar to 17β-estradiol. It has antioxidant properties and a high affinity for estrogen receptors. It can inhibit the activity of tyrosine protein kinase (PTK) and topoisomerase II, and has the effects of inducing programmed cell death, enhancing the efficacy of anticancer drugs, and inhibiting angiogenesis. It is a type of flavonoid, and its chemical structure is as follows.

[0005]

[0006] Studies have shown that genistein can induce apoptosis in human breast cancer cells MDA-MB-231 (In vitro study on NF-κB and MAPK signaling pathways involved in genistein-induced apoptosis in breast cancer MDA-MB-231 cells, Chinese Journal of Pharmacology, Vol. 34, No. 5, 2018).

[0007] α-Solanine (C 45 H 73 NO 15 It was first discovered in black nightshade (Solanum nigrum L.), also known as solanine or glycoalkaloid. Later, it was also isolated from potato (Solanum tuberosum Linn.) of the Solanaceae family. Its chemical structure is as follows.

[0008]

[0009] Studies have shown that α-solanine can affect and reduce the expression of proteins on tumor cell membranes, such as reducing sialic acid content, inhibiting ATPase activity, and downregulating N-acetyltransferase expression, thereby reducing tumor cell membrane fluidity and inhibiting the metabolic growth of tumor cells. Solanine can also reduce energy metabolism within cancer cells, inhibiting their growth and proliferation. Simultaneously, solanine can activate apoptosis signaling pathways in cancer cells, promoting apoptosis. Furthermore, solanine can downregulate vascular endothelial growth factor and inhibit tumor angiogenesis (Research Progress and Clinical Application of Solanine's Antitumor Effects, Master's Thesis, Hebei Medical University, No. 04, 2017).

[0010] An unexpected discovery was made during research on the therapeutic effects and mechanisms of genistein and α-solanine on breast cancer, leading to this invention. Summary of the Invention

[0011] The purpose of this invention is to provide a traditional Chinese medicine monomer composition and its use in preparing a drug for treating breast cancer.

[0012] The above-mentioned objective of this invention is achieved through the following technical solution:

[0013] A single-component composition of traditional Chinese medicine, consisting of genistein and α-solanine.

[0014] Preferably, the molar ratio of genistein to α-solanine in the herbal monomer composition is 10:1.

[0015] The above-mentioned traditional Chinese medicine monomer composition is used in the preparation of drugs for treating breast cancer.

[0016] Beneficial effects:

[0017] Those skilled in the art know that genistein and α-solanine possess antitumor activity individually. During the study of the antitumor mechanism of these two components, the researchers of this invention unexpectedly discovered a synergistic effect when used together to treat breast cancer. This discovery led to the formulation of this invention. The synergistic effect revealed by the results of this invention makes genistein and α-solanine a promising candidate for development into a combination therapy for breast cancer. Attached Figure Description

[0018] Figure 1 The inhibition rate of different concentrations of genistein on the proliferation of human breast cancer cells;

[0019] Figure 2 The inhibition rate of human breast cancer cell proliferation by different concentrations of α-solanine. Detailed Implementation

[0020] The following describes the substantive content of the present invention in detail with reference to embodiments, but this does not limit the scope of protection of the present invention.

[0021] Example 1:

[0022] 1. Materials

[0023] The human breast cancer MCF-7 cells were cryopreserved at Jiangsu Cancer Hospital and were used after being thawed using standard methods.

[0024] L-glutamine high-glucose DMEM medium and fetal bovine serum were purchased from Gibco, and the CCK8 kit was purchased from Suzhou Youyilandi Biotechnology Co., Ltd.

[0025] Gentian isoflavone and α-solanine were purchased from Shanghai Yuanye Biotechnology Co., Ltd. with a purity of not less than 98%.

[0026] Both antibiotics and DMSO are standard cell culture reagents.

[0027] 2. Method

[0028] 2.1 Cell Culture

[0029] Human breast cancer MCF-7 cells were cultured in high-glucose DMEM medium containing 10% fetal bovine serum and 200 mM L-glutamine in a constant-temperature carbon dioxide incubator at 37°C and 5% CO2, while maintaining a certain level of humidity. Cell passage was performed by digestion with 0.25% trypsin.

[0030] 2.2 Solution Preparation

[0031] Gentian isoflavone stock solution: Dissolve gentian isoflavone in DMSO to prepare a stock solution, and store it in a refrigerator at 4°C for later use.

[0032] α-Solanine stock solution: Dissolve α-solanine in DMSO to prepare a stock solution, and store it in a refrigerator at 4°C for later use;

[0033] Mother liquor of the composition: Dissolve genistein and α-solanine in DMSO at a molar ratio of 10:1 to prepare a mother liquor, and store at 4°C for later use.

[0034] 2.3 CCK8 assay to detect the inhibitory effect of drugs on the proliferation of MCF-7 breast cancer cells

[0035] Cell suspensions were prepared, cells were counted, and the cell density was adjusted to 30,000 / ml. Cells were seeded into 96-well plates, 100 μl of cell suspension per well. Cells were cultured overnight at 37°C. The next day, different concentrations of the drug (as shown in Table 1) were added for treatment, 100 μl per well. After 48 h of culture, the liquid in the 96-well plates was aspirated, and diluted CCK8 working solution (9 volumes of serum-free medium: 1 volume of CCK8) was added, 100 μl per well. After incubation for 1–2 h, the absorbance was measured at 450 nm using a microplate reader. The growth inhibition rate of each group of cells was calculated, and inhibition rate curves were plotted.

[0036] Table 1. Concentration gradient settings for different drugs

[0037]

[0038] 2.4 Data Analysis and Collaborative Discriminant Analysis

[0039] The results were analyzed using SPSS 16.0. The experiment was repeated three times, and the data are expressed as mean ± s. Co-discrimination was performed using the Kim Jong-jun q-value method: q = E a+b / (E a +E b -E a ×E b E a and E b The inhibition rates of genistein alone and α-solanine alone were E, respectively. a+bThe denominator represents the inhibition rate of the composition; the numerator represents the "measured combined effect", the denominator is the "expected combined effect", the q value is the ratio of the two, q < 0.85 is the antagonistic effect, 0.85 ≤ q < 1.15 is the additive effect, and q ≥ 1.15 is the synergistic effect.

[0040] 3. Results

[0041] The inhibition rates of different drug concentrations on the proliferation of human breast cancer MCF-7 cells are shown in Table 2 and Figures 1-2 As shown.

[0042] Table 2. Inhibition rate of different drug concentrations on the proliferation of human breast cancer MCF-7 cells

[0043]

[0044] The combined drug efficacy q values ​​were all greater than 1.15, indicating a synergistic effect. Those skilled in the art know that genistein and α-solanine possess antitumor activity individually. The researchers of this invention unexpectedly discovered a synergistic effect when these two components are used in combination to treat breast cancer while studying their antitumor mechanism. This discovery led to the proposal of this invention. The synergistic effect revealed by the results of this invention makes genistein and α-solanine a promising candidate for development into a combination therapy for breast cancer.

[0045] The purpose of the above embodiments is to specifically illustrate the substantive content of the present invention, but those skilled in the art should know that the scope of protection of the present invention should not be limited to the specific embodiments.

Claims

1. The use of a traditional Chinese medicine monomer composition for preparing a drug for treating breast cancer, wherein the traditional Chinese medicine monomer composition is composed of genistein and α-solanine, and the molar ratio of genistein to α-solanine is 10:1.

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