Application of oritavancin in preparation of medicine for treating breast cancer

Abstract

The invention relates to the technical field of medicines, in particular to application of oritavancin in preparation of a medicine for treating breast cancer. The invention provides application of oritavancin in preparation of a medicine for treating breast cancer. Experiments prove that oritavancin has excellent potential for preparing the medicine for treating the breast cancer, and oritavancin can inhibit transverse migration movement ability, clone formation ability and multiplication ability of triple-negative breast cancer cells and has a relatively good application prospect.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly to the application of oritavancin in the preparation of drugs for treating breast cancer. Background Art

[0002] Triple-negative breast cancer (TNBC) is a subtype of breast cancer with high malignancy and poor prognosis; the hormone receptor and human epidermal growth factor receptor 2 are both negatively expressed, resulting in its non-responsiveness to endocrine therapy and targeted therapy, which is the key reason for its poor prognosis.

[0003] Chemotherapy is currently the most important systemic treatment for triple-negative breast cancer. However, currently commonly used chemotherapeutic drugs, such as paclitaxel, cyclophosphamide, platinum-based drugs, anthracyclines, etc., have the disadvantages of few types, high toxicity, easy drug resistance, and slow drug iteration, with no update in the past 10 years. Although immunotherapy, anti-angiogenic therapy, PARP inhibitors, etc. have made certain progress, their indications and efficacy are limited. The current situation of the lack of drugs for TNBC remains a clinical bottleneck problem restricting the survival and quality of life of patients.

[0004] Oritavancin is a glycopeptide antibiotic that has a bactericidal effect on various Gram-positive bacteria such as staphylococci, streptococci, and enterococci. Its antibacterial mechanism is similar to that of other glycopeptide antibiotics: it inhibits the synthesis of peptidoglycan in the bacterial cell wall by inhibiting transglycosylation; oritavancin contains both lipophilic and hydrophilic groups, and the additional lipophilic side chain helps to anchor the drug on the target cell membrane, thereby enhancing the effect; its hydrophobic 4'-chlorobiphenylmethyl group can interact with the bacterial cell membrane, and disrupt the bacterial cell membrane by depolarization and increasing cell membrane permeability, resulting in rapid cell death. Clinical trials have demonstrated the safety of oritavancin. Its maximum tolerated dose is 1200 mg, the half-life is 245 hours, and the common adverse events are gastrointestinal reactions, including diarrhea, nausea, vomiting, as well as dizziness, headache, and phlebitis at the infusion site. Taking corresponding symptomatic treatment measures can effectively control the symptoms. Currently, oritavancin has completed clinical trials and has been approved by the FDA for the treatment of bacterial skin infections caused by Gram-positive bacteria (including Staphylococcus aureus, Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus anginosus, and Enterococcus faecalis, etc.).

[0005] In addition to its antibacterial function, oritavancin has also been found to be a non-specific weak inhibitor of multiple cytochrome P isoenzymes (CYP2C9 and CYP2C19), and a non-specific inducer of several other cytochrome P isoenzymes (CYP3A4 and CYP2D6). Given the potential association between cytochrome P isoenzymes and the occurrence and development of tumors, these complex mechanisms of oritavancin are likely to also inhibit tumor cells, but currently, there are no relevant studies on the effect of oritavancin on tumor growth. Summary of the Invention

[0006] The object of the present invention is to provide the use of oritavancin in the preparation of a medicament for treating breast cancer.

[0007] In order to achieve the above object of the invention, the present invention provides the following technical solutions:

[0008] The present invention provides the use of oritavancin in the preparation of a medicament for treating breast cancer.

[0009] Preferably, the breast cancer is triple-negative breast cancer.

[0010] Preferably, the pathogenic cancer cells of the breast cancer are MDA-MB-231 or 4T1.

[0011] Preferably, the medicament for treating breast cancer is a preparation having the ability to inhibit the lateral migration movement of cancer cells.

[0012] Preferably, the medicament for treating breast cancer is a preparation having the ability to inhibit the colony formation ability of cancer cells.

[0013] Preferably, the medicament for treating breast cancer is a preparation having the ability to inhibit the proliferation ability of cancer cells.

[0014] Preferably, the medicament for treating breast cancer is a preparation having the ability to kill cancer cells.

[0015] The present invention also provides a medicament for treating breast cancer, which contains oritavancin and excipients.

[0016] Preferably, the concentration of oritavancin in the medicament for treating breast cancer is 3-130 μM.

[0017] Preferably, the dosage form of the medicament is capsule, pill, tablet, granule or injection.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] 1. Through in vitro cell experiments, the present invention finds that oritavancin has excellent potential for preparing a medicament for treating breast cancer. Oritavancin can inhibit the lateral migration movement ability, colony formation ability and proliferation ability of triple-negative breast cancer cells, and has a good application prospect. And the killing effect of oritavancin on TNBC is not inferior to that of the existing TNBC treatment drug cyclophosphamide.

[0020] 2. In the prior art, oritavancin is used as a mature antibiotic and its human safety has been confirmed through clinical trials. Compared with other preclinical drugs, its toxicity is known and controllable. Through in-depth research on the anti-cancer mechanism of oritavancin, the present invention can provide a theoretical basis for the research and development of new drug action targets for TNBC and promote the further research of TNBC drugs. Description of the Drawings

[0021] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the drawings in the following description are only the embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained according to the provided drawings.

[0022] Figure 1 It is a survival rate graph of MDA-MB-231 cells and 4T1 cells.

[0023] Figure 2 It is a colony formation experiment graph of MDA-MB-231 cells and 4T1 cells.

[0024] Figure 3 It is the colony formation rate of MDA-MB-231 cells and 4T1 cells.

[0025] Figure 4 It is a cell scratch repair experiment graph of MDA-MB-231 cells and 4T1 cells. Specific Embodiments

[0026] The following will combine the embodiments to elaborate on the technical solutions provided by the present invention in detail, but they cannot be understood as limiting the protection scope of the present invention.

[0027] Example 1

[0028] I. Experiment to Evaluate the Effect of Oritavancin on the Growth of TNBC Cells

[0029] 1. CCK-8 Experiment

[0030] ① Take two TNBC cells in the logarithmic growth phase: MDA-MB-231 and 4T1, and prepare cell suspensions of 5×10 4 cells / ml respectively, evenly plate them on 96-well plates, add a circle of PBS solution around, and after 24 hours, the cells adhere and grow.

[0031] ② Add different concentrations of oritavancin (concentration gradients are 0, 5, 10, 25, 50, 100, 250, 500 μM), after treatment for 12 / 24 / 36 / 48 hours, wash the wells with PBS, add 100 μl of complete medium containing 10% CCK-8 reagent, and incubate at 37°C for 2 hours.

[0032] ③ Measure the absorbance (OD value) at 450 nm using an enzyme-linked immunosorbent assay (ELISA) reader. The absorbance of the cell-free blank well is Ab, the absorbance of the control well with a drug concentration of 0 is Ac, and the absorbance of the experimental well is As. The formula for calculating cell viability is [(As - Ab) / (Ac - Ab)] × 100%. Plot the growth curve and calculate the half-maximal inhibitory concentration (IC50) value of the tumor cells.

[0033] 2. Colony formation assay

[0034] ① Select an appropriate concentration of oritavancin based on the results of the CCK-8 assay. Prepare a suspension of MDA-MB-231 cells at 3×10 3 / ml and a suspension of 4T1 cells at 1×10 3 / ml. Pipette 1 ml of each suspension into separate wells of a 6-well plate containing 1 ml of complete medium and mix well.

[0035] ② After 24 hours, when the cells have adhered and grown, replace the medium of the MDA-MB-231 cells with complete medium, 30 μM oritavancin solution, or 10 μl of DMSO; replace the medium of the 4T1 cells with complete medium, 80 μM oritavancin solution, or 30 μl of DMSO.

[0036] ③ After 24 hours of treatment, wash the cells with PBS and replace the medium with complete medium. Observe the cell growth and replace the medium regularly until cell clusters are visible to the naked eye.

[0037] ④ Wash the cells with PBS. Add 200 μl of 4% tissue cell fixative to each well and fix at room temperature for 30 minutes. Then wash with PBS. Add 200 μl of crystal violet stain to each well and stain at room temperature for 30 minutes. Wash the 6-well plate, air dry, take a photo, and count the cell colonies using Image J software. Calculate the cell colony formation rate.

[0038] II. Experiment to evaluate the effect of oritavancin on the migration ability of TNBC cells

[0039] Cell scratch repair assay

[0040] ① Use a marker pen to draw three parallel lines on the bottom of a 6-well plate as a mark. Prepare a suspension of MDA-MB-231 and 4T1 cells at 1×10 6 / ml. Add 2 ml of each cell suspension to each well of the 6-well plate.

[0041] ② After 24 hours, use a 200 μl yellow pipette tip to gently and quickly draw a straight line perpendicular to the above-mentioned marked line at the bottom of the well. After washing off the detached cells with PBS, replace the medium with 2 ml of FBS-free medium containing oritavancin (at concentrations of 15 and 30 μM for MDA-MB-231; at concentrations of 35 and 80 μM for 4T1). Take a microscopic photo to record the cell healing situation at the upper edge of the intersection of the scratch and the marked line.

[0042] ③ After 24 / 48 hours, wash the 6-well plates with PBS and add fresh serum-free medium. Take pictures at the same position to record the cell healing condition, and analyze and calculate the scratch repair rate with Image J software and draw a graph.

[0043] Results:

[0044] 1. The results of the CCK-8 assay are as Figure 1 shown. The IC50 values of oritavancin for MDA-MB-231 cells at 12 hours, 24 hours, 36 hours, and 48 hours were 37.55, 34.22, 10.27, and 3.93 μM, respectively; the IC50 values of oritavancin for 4T1 cells at 12 hours, 24 hours, 36 hours, and 48 hours were 125.29, 84.11, 47.27, and 27.50 μM, respectively. Oritavancin could significantly inhibit the proliferation of MDA-MB-231 cells and 4T1 cells, and the cell survival rate showed an obvious downward trend. Moreover, with the prolongation of the drug action time, the inhibitory effect was significantly enhanced.

[0045] 2. The results of the colony formation assay are as Figure 2 and 3 shown. In MDA-MB-231 cells, the colony formation rate of the oritavancin treatment group was 34.10%, that of the cyclophosphamide treatment group was 28.73%, that of the DMSO solvent control group was 47.80%, and that of the blank control group was 53.23%; in 4T1 cells, the colony formation rate of the oritavancin treatment group was 7.30%, that of the cyclophosphamide treatment group was 31.03%, that of the DMSO solvent control group was 55.20%, and that of the blank control group was 62.37%. Oritavancin significantly inhibited the colony formation ability of MDA-MB-231 cells and 4T1 cells, and this inhibitory effect was better than that of the traditional anti-TNBC drug cyclophosphamide.

[0046] 3. The results of the cell scratch repair assay are as Figure 4As shown, the 24-hour cell scratch repair rate of MDA-MB-231 cells was 20.01% in the blank control group, 8.22% in the oritavancin 15 μM group, and 6.32% in the oritavancin 30 μM group. The 48-hour cell scratch repair rate was 35.38% in the blank control group, 20.47% in the oritavancin 15 μM group, and 11.57% in the oritavancin 30 μM group; the 24-hour cell scratch repair rate of 4T1 cells was 18.10% in the blank control group, 8.58% in the oritavancin 35 μM group, and -2.98% in the oritavancin 80 μM group; the 48-hour cell scratch repair rate was 29.64% in the blank control group, 13.81% in the oritavancin 35 μM group, and -8.63% in the oritavancin 80 μM group. In MDA-MB-231 cells and 4T1 cells, the cell scratch repair rate after oritavancin treatment was significantly decreased and showed a concentration-dependent manner. In the high-dose oritavancin group, due to excessive cell death, there was even a tendency for the scratch to increase. Oritavancin significantly inhibited the lateral migration ability of MDA-MB-231 and 4T1 cells.

[0047] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. The use of oritavancin in the preparation of drugs for the treatment of breast cancer.

2. The use according to claim 1, characterized in that: The breast cancer is triple-negative breast cancer.

3. The use according to claim 1 or 2, characterized in that: The pathogenic cancer cells of breast cancer are MDA-MB-231 or 4T1.

4. The use according to claim 1 or 2, characterized in that: The drug for treating breast cancer is a preparation having the ability to inhibit the lateral migration of cancer cells.

5. The use according to claim 1 or 2, characterized in that: The drug for treating breast cancer is a preparation having the ability to inhibit the formation of cancer cell clones.

6. The use according to claim 1 or 2, characterized in that: The drug for treating breast cancer is a preparation having the ability to inhibit the proliferation of cancer cells.

7. The use according to claim 1 or 2, characterized in that: The drug for treating breast cancer is a preparation having the ability to disinfect cancer cells.

8. A drug for treating breast cancer, characterized in that: The medicine contains oritavancin and excipients.

9. The drug according to claim 8, characterized in that The effective concentration of oritavancin in the drug for treating breast cancer is 3 to 130 μM.

10. The drug according to claim 8, characterized in that The dosage form of the medicine is capsule, pill, tablet, granule or injection.

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