Application of Lactobacillus fermentum MSJK0025 derived from breast milk in the prevention and / or treatment of breast cancer

Abstract

This application relates to the application of a strain of *Lactobacillus fermentum* MSJK0025 derived from breast milk in the prevention and / or treatment of breast cancer. Specifically, it relates to the use of *Lactobacillus fermentum* MSJK0025 derived from breast milk in the preparation of pharmaceutical compositions for the prevention and / or treatment of breast cancer, and the use of *Lactobacillus fermentum* MSJK0025 derived from breast milk in the preparation of probiotic agents for the prevention and / or treatment of breast cancer. This application provides a novel application of a strain of *Lactobacillus fermentum* MSJK0025 derived from breast milk in the prevention and / or treatment of breast cancer, utilizing *Lactobacillus fermentum* MSJK0025 to effectively slow the early growth of breast tumors. Therefore, *Lactobacillus fermentum* MSJK0025 can be well applied in the prevention and / or treatment of breast cancer, especially in the treatment of early-stage breast cancer, thereby improving breast health.

Description

Technical Field

[0001] This application relates to the field of microbial technology, and in particular to the use of a breast milk-derived Lactobacillus fermentum MSJK0025 in the prevention and / or treatment of breast cancer. Background Technology

[0002] Breast cancer is one of the most common malignant tumors in women, typically occurring in the glandular epithelial tissue of the breast. Statistics show that its incidence accounts for 7%-10% of all malignant tumors. Its development is often related to genetics, and the incidence is higher in women aged 40-60, particularly around menopause. It is one of the most common malignant tumors that seriously affects women's physical and mental health and can even be life-threatening.

[0003] Currently, the main treatments for breast cancer are surgery and Western medicine. However, surgery carries certain risks and is not suitable for older or weaker patients; Western medicine often only treats the symptoms and not the root cause, with limited effectiveness and high cost. Therefore, there is a need to find new methods for improving or treating breast cancer.

[0004] The *Lactobacillus fermentum* MSJK0025 involved in this application was also isolated from the breast milk of healthy postpartum women. Studies have shown that *Lactobacillus fermentum* MSJK0025 exhibits superior resistance to gastric acid and bile salts, excellent intestinal colonization ability, and high growth rate. Furthermore, it has completed safety evaluation tests and commercial batch production. Research data indicates that *Lactobacillus fermentum* MSJK0025 possesses advantages such as strong tolerance, good safety, and low production cost, demonstrating promising industrialization prospects. Simultaneously, research data shows that *Lactobacillus fermentum* MSJK0025 has a good effect on improving mastitis and thus breast health; however, its effects and efficacy in preventing, improving, or treating breast cancer require further investigation. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this application provides the application of a breast milk-derived Lactobacillus fermentum MSJK0025 in the prevention of breast cancer. Lactobacillus fermentum MSJK0025 can effectively slow down the early growth of breast tumors, thereby effectively preventing the occurrence of breast cancer in female mammals, including humans, and treating and alleviating early breast cancer in female mammals, including humans.

[0006] Therefore, the first aspect of this application provides a strain of Lactobacillus fermentum derived from breast milk (… Limosilactobacillus fermentum The use of MSJK0025 in the preparation of pharmaceutical compositions for the prevention and / or treatment of breast cancer.

[0007] The *Lactobacillus fermentum* MSJK0025 used in this application is the strain isolated and screened in patent application number 202311070808.2, which is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 26426. Through research on this strain, the inventors of this application have discovered that it can effectively alleviate the early growth of breast tumors, and therefore can be effectively used in pharmaceutical compositions for the prevention and / or treatment of breast cancer.

[0008] In addition to *Lactobacillus fermentum* MSJK0025, the pharmaceutical composition of this application may also include a pharmaceutically acceptable carrier. In this application, the pharmaceutically acceptable carrier includes, but is not limited to, any pharmaceutically acceptable excipient, surfactant, desiccant, diluent, carrier, adjuvant, or additive. Suitable pharmaceutically acceptable carriers may include, for example, magnesium carbonate, lactose, pectin, dextrin, starch, astragalus gum, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, and cocoa butter. In this application, the pharmaceutically acceptable carrier may be solid, semi-solid, or liquid.

[0009] In some embodiments, the pharmaceutical composition is administered to female mammals, including humans.

[0010] The *Lactobacillus fermentum* MSJK0025 in the pharmaceutical composition of this application can effectively slow down the early growth of breast tumors. Specifically, in the experimental group treated with *Lactobacillus fermentum* MSJK0025, the tumor volume of female mice with breast cancer was significantly reduced at week 13, and the tumor weight was significantly reduced at both week 13 and week 17. Therefore, the pharmaceutical composition of this application can effectively prevent or treat breast cancer, and thus can be effectively applied to female mammals, including humans.

[0011] The mammals mentioned in this application may be, for example, humans, rats, pigs, cattle, sheep, dogs, cats, rabbits, etc.

[0012] In some embodiments, the pharmaceutical composition is used to treat early-stage breast cancer.

[0013] In this application, early breast cancer (or early-stage breast cancer) refers to a patient whose tumor is less than 3cm during surgery; no metastatic lymph nodes are palpable in the axilla, and there is no distant metastasis, or the ipsilateral axillary lymph nodes have no metastasis or only micrometastasis, and no distant metastasis; including non-invasive cancer.

[0014] The *Lactobacillus fermentum* MSJK0025 in this application can slow down the early growth of breast tumors (the tumor volume of female mice with breast cancer in the experimental group treated with *Lactobacillus fermentum* MSJK0025 was significantly reduced at week 13, and the tumor weight was significantly reduced at both weeks 13 and 17), but it does not improve the malignant development of breast tumors. Therefore, the breast cancer treated by the pharmaceutical composition described in this application is preferably early-stage breast cancer.

[0015] The second aspect of this application provides the use of a breast milk-derived Lactobacillus fermentum MSJK0025 in the preparation of a probiotic agent for the prevention and / or treatment of breast cancer.

[0016] The *Lactobacillus fermentum* MSJK0025 used in this application is the strain isolated and screened in patent application number 202311070808.2, which is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 26426. Through research on this strain, the inventors of this application have discovered that it can effectively alleviate the early growth of breast tumors, and therefore can be effectively used in probiotic preparations for the prevention and / or treatment of breast cancer.

[0017] In addition to Lactobacillus fermentum MSJK002, the probiotic agent in this application may also include other probiotics, and this application does not limit them.

[0018] In some embodiments, the probiotic agent is administered to female mammals, including humans.

[0019] The probiotic agent of this application contains *Lactobacillus fermentum* MSJK0025, which can effectively slow down the early growth of breast tumors. Specifically, in the experimental group treated with *Lactobacillus fermentum* MSJK0025, the tumor volume of female mice with breast cancer was significantly reduced at week 13, and the tumor weight was significantly reduced at both week 13 and week 17. Therefore, the probiotic agent of this application can effectively prevent or treat breast cancer, and can be effectively applied to female mammals, including humans.

[0020] The mammals mentioned in this application may be, for example, humans, rats, pigs, cattle, sheep, dogs, cats, rabbits, etc.

[0021] In some embodiments, the probiotic agent is used to treat early-stage breast cancer.

[0022] In this application, early breast cancer (or early-stage breast cancer) refers to a patient whose tumor is less than 3cm during surgery; no metastatic lymph nodes are palpable in the axilla, and there is no distant metastasis, or the ipsilateral axillary lymph nodes have no metastasis or only micrometastasis, and no distant metastasis; including non-invasive cancer.

[0023] The *Lactobacillus fermentum* MSJK0025 in this application can slow down the early growth of breast tumors (the tumor volume of female mice with breast cancer in the experimental group treated with *Lactobacillus fermentum* MSJK0025 was significantly reduced at week 13, and the tumor weight was significantly reduced at both weeks 13 and 17), but it has no effect on improving the malignant development of breast tumors. Therefore, the breast cancer treated with the probiotic agent described in this application is preferably early-stage breast cancer.

[0024] A third aspect of this application provides a pharmaceutical composition for the prevention and / or treatment of breast cancer, the pharmaceutical composition containing *Lactobacillus fermentum* MSJK0025.

[0025] In addition to *Lactobacillus fermentum* MSJK0025, the pharmaceutical composition of this application may also include a pharmaceutically acceptable carrier. In this application, the pharmaceutically acceptable carrier includes, but is not limited to, any pharmaceutically acceptable excipient, surfactant, desiccant, diluent, carrier, adjuvant, or additive. Suitable pharmaceutically acceptable carriers may include, for example, magnesium carbonate, lactose, pectin, dextrin, starch, astragalus gum, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, and cocoa butter. In this application, the pharmaceutically acceptable carrier may be solid, semi-solid, or liquid.

[0026] The fourth aspect of this application provides a probiotic preparation for the prevention and / or treatment of breast cancer, said probiotic preparation containing *Lactobacillus fermentum* MSJK0025.

[0027] In addition to live bacteria of Lactobacillus fermentum MSJK0025, the probiotic agent in this application may also include other probiotics, and this application does not limit it.

[0028] The beneficial technical effects of this application are as follows: This application provides a novel application of *Lactobacillus fermentum* MSJK0025, derived from breast milk, in the prevention and / or treatment of breast cancer. In the experimental group treated with *Lactobacillus fermentum* MSJK0025, the tumor volume of female mice with breast cancer significantly decreased at week 13, and the tumor weight significantly decreased at both weeks 13 and 17. Therefore, *Lactobacillus fermentum* MSJK0025 can be effectively used in the prevention and / or treatment of breast cancer, especially in the treatment of early-stage breast cancer, thereby improving breast health. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 It is a comparison chart of the tumor volumes of mice in the model control group and the Lactobacillus mucosae fermentum group in Example 1.

[0030] Figure 2 It is a comparison chart of the tumor weights of mice in the model control group and the Lactobacillus mucosae fermentum group in Example 1.

[0031] Figure 3 It is an external view of the whole-body tumors of mice in the model control group and the Lactobacillus mucosae fermentum group in Example 1; where A represents the model control group and B represents the Lactobacillus mucosae fermentum group.

[0032] BIOLOGICAL DEPOSIT

[0033] Lactobacillus mucosae fermentum ( Limosilactobacillus fermentum ) MSJK0025, with the deposit number CGMCC No.26426, deposited with the China General Microbiological Culture Collection Center (abbreviation: CGMCC, address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing), and the deposit date is January 9, 2023. SPECIFIC EMBODIMENTS

[0034] To make the present application easier to understand, the following will further elaborate on the present application in combination with examples. These examples are only illustrative and are not limited to the application scope of the present application. Raw materials or components used in the present application can be obtained through commercial channels or conventional methods without special instructions.

[0035] The experimental animals in the following examples are MMTV-PyMT transgenic mice, female, 6 weeks old. The experimental animals are provided by Jiangsu Huachuang Xinnuo Pharmaceutical Technology Co., Ltd., certificate number: SCXK (Su) 2020-0009.

[0036] Example 1: Evaluation of the efficacy of Lactobacillus mucosae fermentum MSJKOO25 in preventing and treating breast cancer

[0037] 1. Animal grouping and administration

[0038] Take MMTV-PyMT transgenic mice (female, 6 weeks old). After the adaptive feeding is completed, the mice are randomly divided into two groups, namely the model control group and the Lactobacillus mucosae fermentum group, with 32 mice in each group, for a total of 64 mice.

[0039] Starting at 6 weeks of age, the model control group and the *Lactobacillus fermentum* group were administered the drug, respectively. The *Lactobacillus fermentum* group was administered the drug 3 times per week, with each mouse receiving 0.5 mL of sample orally via gavage. The model control group was administered the drug 3 times per week, with each mouse receiving 0.5 mL of water orally via gavage. The sample preparation method for the *Lactobacillus fermentum* group was as follows: 20 mg of *Lactobacillus fermentum* MSJK0025 dry powder was weighed and added to 20 ml of water to prepare a bacterial powder solution for later use.

[0040] 2. Efficacy Testing

[0041] When the animals reached 13, 15, 17, and 19 weeks of age, 8 mice from each group were selected for the following tests:

[0042] (1) Weigh yourself;

[0043] (2) Measure the volume of the tumor in the breast with calipers (record 0 if not measured). Since there are multiple diffuse tumors in different locations on the ventral side of the mouse, only the largest tumor near the breast is selected for measurement and recording.

[0044] (3) After euthanizing the mice, remove the tumors from the whole body, weigh and record the tumor weight, and take pictures;

[0045] (4) Blood collection and serum separation: The levels of CA153, VEGF and CEA in serum were detected by ELISA kit.

[0046] 3. Experimental Results

[0047] (1) Expression levels of CA153, VEGF and CEA in mouse serum

[0048] The expression levels of CA153, VEGF, and CEA in mouse serum were detected using the mouse carcinoembryonic antigen (CEA; CD66) ELISA research kit (MM-0633M1, Jiangsu Enzyme Immunosorbent Assay Co., Ltd., China), the mouse vascular endothelial growth factor (VEGF) ELISA research kit (MM-0128M1, Jiangsu Enzyme Immunosorbent Assay Co., Ltd., China), and the mouse carbohydrate antigen 15-3 (CA15-3) ELISA research kit (MM-47737M1, Jiangsu Enzyme Immunosorbent Assay Co., Ltd., China). The results are shown in Table 1.

[0049] Table 1: Expression levels of CA153, VEGF, and CEA in mouse serum (n=8)

[0050]

[0051] Note: Compared with the model control group, * p <0.05,** p <0.01

[0052] As shown in Table 1, compared with the model control group, the expression of VEGF and CEA in the serum of mice in the fermented Lactobacillus group decreased slightly at week 13, with the decrease in CEA expression being more significant. However, it increased significantly at week 19, and the difference was not significant at other times.

[0053] (2) Tumor volume in mice

[0054] The tumor volumes of mice in the model control group and the fermented *Lactobacillus mucinus* group are shown in Table 2. Figure 1 As shown.

[0055] Table 2: Results of tumor volume detection in mice (n = 8)

[0056]

[0057] Note: Compared with the model control group, * p <0.05,** p <0.01

[0058] From Table 2 and Figure 1 The results showed that, compared with the model control group, the tumor volume of mice in the *Lactobacillus fermentum* group was significantly smaller at week 13, with no significant difference at other times. This indicates that administration of *Lactobacillus fermentum* MSJK0025 can slow the growth of early-stage breast tumors.

[0059] (3) Mouse tumor weight

[0060] The tumor weights of mice in the model control group and the fermented *Lactobacillus mucinus* group are shown in Table 3. Figure 2 As shown, and the whole-body tumor images of the mouse are as follows. Figure 3 As shown.

[0061] Table 3: Results of tumor weight detection in mice (n = 8)

[0062]

[0063] Note: Compared with the model control group, * p <0.05

[0064] From Table 3 and Figure 2 It was found that, compared with the model control group, the tumor weight of mice in the *Lactobacillus mucinus* fermentation group was significantly reduced at weeks 13 and 17, with no significant difference at other times. Meanwhile, from... Figure 3 The results showed that, compared with the model control group, the tumor weight and tumor size of mice in the *Lactobacillus fermentum* group were significantly reduced at weeks 13 and 17, with no significant differences at other times. This indicates that administration of *Lactobacillus fermentum* MSJK0025 can slow the growth of early-stage breast tumors.

[0065] The above results indicate that *Lactobacillus fermentum* MSJK0025 has the effect of slowing down the early growth of breast tumors, but it does not improve the malignant development of breast tumors. Therefore, *Lactobacillus fermentum* MSJK0025 can be used in the prevention and treatment of breast cancer (especially the treatment of early-stage breast cancer).

[0066] It should be noted that the embodiments described above are only for explaining this application and do not constitute any limitation on this application. This application has been described with reference to typical embodiments, but it should be understood that the terms used therein are descriptive and explanatory terms, not limiting terms. Modifications can be made to this application within the scope of the claims, and revisions can be made to the invention without departing from the scope and spirit of this application. Although the application described herein relates to specific methods, materials, and embodiments, it does not mean that this application is limited to the specific examples disclosed herein; on the contrary, this application can be extended to all other methods and applications with the same function.

Claims

1. The use of Limosilactobacillus fermentum MSJK0025, derived from breast milk, in the preparation of a pharmaceutical composition for the prevention and / or treatment of breast cancer, wherein the breast cancer treated is early-stage breast cancer.

2. The application according to claim 1, characterized in that, The pharmaceutical composition is to be administered to female mammals, including humans.

3. The use of Lactobacillus fermentum MSJK0025 derived from breast milk in the preparation of a probiotic agent for the prevention and / or treatment of breast cancer, wherein the breast cancer treated by the probiotic agent is early-stage breast cancer.

4. The application according to claim 3, characterized in that, The probiotic preparation is intended for use by female mammals, including humans.

Images