Application of norcoclaurine in anti-triple negative breast cancer and its bone metastasis tumor drugs

Abstract

The present application relates to the application of norharmine in anti-triple negative breast cancer and bone metastasis tumor drugs, and relates to the field of medicine. Norharmine can significantly enhance the killing effect of lymphocytes on triple negative breast cancer cells in vitro experiments, and then treat triple negative breast cancer; at the same time, norharmine significantly inhibits tumor size and osteolysis in a bone metastasis bone destruction mouse model induced by triple negative breast cancer cells. It can be seen that norharmine can be used as an anti-triple negative breast cancer drug and an anti-triple negative breast cancer bone metastasis tumor drug, and has a wide application prospect.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to the application of norcodonine in the treatment of triple-negative breast cancer and its bone metastases. Background Technology

[0002] Triple-negative breast cancer (TNC) refers to breast cancer where immunohistochemical staining of the cancer tissue shows negative results for estrogen receptor (ER), progesterone receptor (PR), and the proto-oncogene HER2. This type of breast cancer exhibits unique biological behavior and clinicopathological characteristics compared to other subtypes, and its prognosis is generally worse. Furthermore, because TNC does not express estrogen receptors, progesterone receptors, or HER2, it cannot be treated with endocrine therapy or HER2-targeted therapies, relying solely on chemotherapy. Treatment options are very limited. However, because its cancer cells proliferate faster, are more invasive, and are more prone to early metastasis compared to other breast cancer cell types, chemotherapy resistance is common, and the risk of recurrence is significantly higher than in other breast cancer subtypes.

[0003] Meanwhile, metastasis is one of the most serious complications of cancer and a leading cause of death for patients with various malignant tumors. Bone tissue is one of the most common sites of metastasis for malignant tumors such as breast cancer, lung cancer, and prostate cancer, especially in breast cancer, where the probability of bone metastasis is as high as 70%. After tumors metastasize to the bone, they can cause a series of bone-related events such as osteolytic bone destruction, bone pain, and fractures, which significantly reduce the patient's quality of life and shorten their survival time. Triple-negative breast cancer bone metastases are more difficult to treat than bone metastases of other breast cancer subtypes. Because triple-negative breast cancer cannot be treated with endocrine or targeted drugs, there is a lack of long-term maintenance regimens. Furthermore, triple-negative cancer cells release more osteoclast activating factors, resulting in more severe bone destruction. Therefore, triple-negative breast cancer bone metastases pose a higher risk of pain, fractures, and spinal cord compression.

[0004] Current cancer treatment methods mainly include surgery, radiotherapy, chemotherapy, immunotherapy, and targeted drug therapy. Among them, immunotherapy has become an important treatment strategy. Its core lies in strengthening the immune system's ability to specifically recognize and attack tumor cells and / or removing the inhibitory effect of tumor cells on the immune system, thereby achieving the effect of treating tumors.

[0005] Traditional Chinese medicine (TCM) has demonstrated significant preventative and therapeutic effects at different stages of cancer, and an increasing number of active ingredients from TCM are being developed and showing good efficacy. Norcodonine, derived from the traditional Chinese medicine Lindera strychnifolia, enhances myocardial contractility and dilates blood vessels in the heart, brain, and peripheral systems. Clinically, it is used to treat chronic arrhythmias and sick sinus syndrome. The structure of norcodonine is as follows:

[0006] Currently, there are no literature reports on the therapeutic effects of norcodonine on triple-negative breast cancer or bone metastases formed from triple-negative breast cancer. Summary of the Invention

[0007] To address the aforementioned problems, this invention provides the application of norcodonol in the preparation of drugs against triple-negative breast cancer. Norcodonol can significantly enhance the killing effect of lymphocytes on 4T1 breast cancer cells (the 4T1 cell line is specifically used for triple-negative breast cancer modeling) in in vitro experiments, thereby inhibiting triple-negative breast cancer cells and their metastatic ability.

[0008] This invention provides the application of norcodonine in the preparation of drugs for treating triple-negative breast cancer.

[0009] In one embodiment, the norcodonine enhances the killing effect of lymphocytes on triple-negative breast cancer cells, thereby achieving anti-triple-negative breast cancer.

[0010] In one embodiment, the triple-negative breast cancer cells include breast cancer 4T1 cells.

[0011] In one embodiment, the chemical structural formula of the norcodonine is shown in Formula I:

[0012] Formula I.

[0013] In one embodiment, the drug further includes a pharmaceutically acceptable carrier and / or excipients.

[0014] This invention also provides the application of norcodonine in the preparation of a drug for treating bone metastases from triple-negative breast cancer.

[0015] In one embodiment, the chemical structural formula of the norcodonine is shown in Formula I:

[0016] Formula I.

[0017] In one embodiment, the drug further includes a pharmaceutically acceptable carrier and / or excipients.

[0018] In one embodiment, the drug is in the form of an oral dosage form or a topical dosage form.

[0019] In one embodiment, the oral dosage form includes tablets, capsules, oral liquids, or granules.

[0020] In one embodiment, the topical dosage form includes injections, lyophilized powders, granules, pills, or ointments.

[0021] In one embodiment, the route of administration of the drug includes oral, sublingual, intravenous, or transdermal administration.

[0022] Compared with the prior art, the present invention has the following beneficial effects: The present invention relates to the application of norcodonoline in the treatment of triple-negative breast cancer and its bone metastases. Norcodonoline significantly enhances the killing effect of lymphocytes on 4T1 breast cancer cells (the 4T1 cell line is specifically used for triple-negative breast cancer modeling) in vitro, thereby inhibiting triple-negative breast cancer cells and their metastatic ability. Simultaneously, in a mouse model of bone metastasis and bone destruction induced by 4T1 breast cancer cells, norcodonoline significantly inhibits bone tumor size and effectively prevents the bone resorption it induces. Therefore, norcodonoline has broad application prospects as an anti-triple-negative breast cancer drug and an anti-triple-negative breast cancer bone metastasis drug. Attached Figure Description

[0023] Figure 1 The figure shows the results of the experiment on the effect of norcodonol on the growth of triple-negative breast cancer cells co-cultured with mouse lymphocytes (Note: compared with the control group, ### p<0.001, p<0.05).

[0024] Figure 2 The figure shows the experimental results of norcodonopsis pilosula inhibiting tumor size formation in triple-negative breast cancer cells in vivo. Figure 2 A shows photographs of the hind limbs on the modeling side for each group. Figure 2 B is a statistical chart of bone tumor area; Zol is the positive control drug zoledronic acid; (Note: compared with the control group) #### p<0.0001, p<0.0001).

[0025] Figure 3 The figure shows the experimental results of norcodonopsis pilosula in improving osteolysis induced by triple-negative breast cancer cells in vivo. Figure 3 A represents the scan images of each group. Figure 3 B is a statistical chart of each indicator; Zol is the positive control drug zoledronic acid; BMD (mg HA / ccm) is the bone mineral density parameter; BV / TV (%) is the bone volume parameter; Tb.N ( / mm) is the number of trabeculae; and Tb.Sp (mm) is the trabecular separation (Note: compared with the control group, ...). #### p<0.0001, p<0.0001, p<0.001, p<0.01). Detailed Implementation

[0026] To facilitate understanding of the present invention, a more complete description will be given below with reference to the accompanying drawings. Preferred embodiments of the invention are shown in the drawings. However, the invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the invention.

[0027] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0028] Unless otherwise specified, all reagents, materials, and equipment used in this embodiment are commercially available; unless otherwise specified, all test methods are conventional test methods in this field.

[0029] source: Mouse spleen lymphocytes (source: 7-8 week old C57 / BL6 female mice); 7-8 week old Balb / c female mice (purchased from Guangdong Yaokang Company); Australian fetal bovine serum (purchased from Gibco, USA); 1640 medium (purchased from Gibco, USA); DMEM medium (purchased from Gibco, USA).

[0030] Example 1 The effect of norcodonine on the growth of triple-negative breast cancer cells co-cultured with mouse lymphocytes in vitro.

[0031] The 4T1 breast cancer cells are derived from Balb / c mouse breast cancer tissue and belong to triple-negative breast cancer (ER-, PR-, HER2-). They are similar in molecular characteristics to human triple-negative breast cancer and are therefore often used to create triple-negative breast cancer models.

[0032] Splenic lymphocytes were obtained from 8-week-old C57 / BL6 female mice and pre-stimulated with anti-CD3 antibody for 24 hours. The stimulated lymphocytes were co-cultured with adherent 4T1 cells at a ratio of 10:1 (lymphocytes:4T1 cells), with 4 μM norcodine added to the co-culture system. The culture was continued for 24 hours, and cancer cell growth was assessed using the MTT assay. Results are as follows: Figure 1 As shown in Figure A.

[0033] Depend on Figure 1 Results A show that norcodonine significantly enhances the killing effect of lymphocytes on 4T1 breast cancer cells in vitro.

[0034] Example 2 Norcodonine inhibits the growth of bone-metastatic triple-negative breast cancer cells in vivo. Animal Experiment Procedure: Eight-week-old female Balb / c mice were randomly divided into four groups (n=5 per group): a negative control group (Sham), a tibial injection group of 4T1 breast cancer cells (Model), a low-dose drug group (Model + HG 10 mg / kg), a high-dose drug group (Model + HG 50 mg / kg), and a positive control group (Model + Zol 0.1 mg / kg). Starting the day after modeling, the negative and positive control groups were administered saline by gavage; the drug intervention groups were administered 10 mg / kg or 50 mg / kg of HG daily by gavage. All mice underwent continuous administration for 21 days before tissue collection. The hind limb from the modeling side was harvested. After removal, the hind limbs were arranged in groups on white paper and photographed. ImageJ software was used to analyze the area, representing tumor size. Results are as follows: Figure 2 As shown in AB.

[0035] Depend on Figure 2 The results of AB studies showed that norcodonine inhibited the growth and tumor size of bone metastatic 4T1 triple-negative breast cancer cells induced by intraosseous injection.

[0036] Example 3 Norcodonine blocks bone destruction induced by triple-negative breast cancer cells in vivo: Eight-week-old female Balb / c mice were randomly divided into four groups: a negative control group (Sham), a tibial injection group of 4T1 breast cancer cells (Model), a low-dose drug group (Model + HG 10 mg / kg), a high-dose drug group (Model + HG 50 mg / kg), and a positive control group (Model + Zol 0.1 mg / kg), with five mice in each group. Starting the day after modeling, the negative and positive control groups were administered saline by gavage; the drug intervention groups were administered 10 mg / kg or 50 mg / kg of HG by gavage daily. All mice were administered the drugs for 21 consecutive days, after which the hind limb from the modeling side was harvested for Micro-CT scanning analysis. Results are as follows: Figure 3 As shown in AB.

[0037] Depend on Figure 3 The results of the AB study showed that norcodonine significantly improved bone destruction induced by 4T1 breast cancer cells in vivo, and the high dose restored bone mass to the level of the healthy control group, while no adverse reactions of bone sclerosis induced by the positive control drug were observed.

[0038] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0039] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. The application of norcodonine in the preparation of drugs for triple-negative breast cancer.

2. The application according to claim 1, characterized in that, Norcodonine enhances the killing effect of lymphocytes on triple-negative breast cancer cells, thereby achieving anti-triple-negative breast cancer.

3. The application according to claim 2, characterized in that, The triple-negative breast cancer cells include triple-negative breast cancer cells from humans and mice.

4. The application according to claim 1, characterized in that, The chemical structural formula of the norcodonine is shown in Formula I: Formula I.

5. The application according to claim 1, characterized in that, The drug also includes pharmaceutically acceptable carriers and / or excipients.

6. The application of norcodonine in the preparation of a drug for treating bone metastases from triple-negative breast cancer.

7. The application according to claim 6, characterized in that, The chemical structural formula of the norcodonine is shown in Formula I: Formula I.

8. The application according to claim 6, characterized in that, The drug also includes pharmaceutically acceptable carriers and / or excipients.

9. The application according to claim 6, characterized in that, The drug is available in oral or topical dosage form.

10. The application according to claim 6, characterized in that, The routes of administration for the drug include oral, sublingual, intravenous, or transdermal.

Images