CN115990147A

CN115990147A - Application of metochalcone in preparation of tumor metastasis inhibiting drugs

Info

Publication number: CN115990147A
Application number: CN202211567723.0A
Authority: CN
Inventors: 彭芙; 周建波
Original assignee: Sichuan University
Current assignee: Sichuan University
Priority date: 2022-12-07
Filing date: 2022-12-07
Publication date: 2023-04-21

Abstract

The invention provides an application of metochalcone in preparing a medicine for inhibiting tumor metastasis, and relates to the field of pharmacy. The invention discovers for the first time that 2',4' -trimethoxychalcone/metochalcone has remarkable effect in inhibiting proliferation of lung cancer and breast cancer. The metochalcone has good application prospect in preparing medicines for inhibiting tumors.

Description

Application of metochalcone in preparation of tumor metastasis inhibiting drugs

Technical Field

The invention provides an application of metochalcone in preparing a medicine for inhibiting tumor metastasis, and relates to the field of pharmacy.

Background

Lung cancer is a leading cause of cancer death worldwide, and at the same time, lung cancer is the first major cancer threatening the health of men worldwide. Lung cancer remains the most common type of cancer in china and the leading cause of cancer death. Lung cancer is classified as non-small cell lung cancer (NSCLC) and Small Cell Lung Cancer (SCLC). Of these, small cell cancers account for about 15% of all lung cancers, the most aggressive, worst prognosis type among all lung cancer cell types, and are markedly correlated with smoking history. Non-small cell lung cancer can be further classified into adenocarcinoma, squamous cell carcinoma and large cell lung cancer. About 70% of lung cancer patients have reached locally advanced stages or metastasized at the time of diagnosis. Breast cancer has become the most common malignant tumor of women worldwide, and the morbidity and mortality rate of the malignant tumor are the first place of the malignant tumor of women, and seriously threaten and endanger the physical and mental health of women. Different types of breast cancer have significantly different biological properties and clinical manifestations, currently, immunohistochemical methods are mainly used clinically, and breast cancer is divided into 4 molecular subtypes according to detection results of Estrogen Receptor (ER), progestogen Receptor (PR) and human epidermal growth factor receptor 2 (HER-2): the luminal A type (ER positive or PR positive, HER-2 negative, ki67 low expression), the luminal B type (ER positive or PR positive, HER-2 positive), the HER-2 over-expression type (ER, PR negative, HER-2 positive, ki67 high expression) and the basal-like type (ER, PR, HER-2 negative). The triple negative breast cancer (triple negative breast cancer, TNBC) is used as a subtype with highest malignancy degree in breast cancer, is characterized by negative ER, PR and HER-2, and has the characteristics of high recurrence rate, high early metastasis rate, strong invasiveness and poor prognosis.

The treatment methods of breast cancer include operation treatment, endocrine treatment, chemotherapy, radiotherapy, immunotherapy and the like, but the methods are difficult to effectively solve the invasion and metastasis problems of cancer cells after breast cancer operation. At present, surgery is the main method for treating breast cancer, but metastasis after cancer cell operation is not rare, 50% -70% of patients generate systemic metastasis within 10 years after simple radical treatment and improved radical treatment, even if primary focus is smaller, axillary lymph nodes clean negative clinical early breast cancer, local metastasis and distant metastasis can also occur early after operation, and prognosis is affected. There are data currently reporting that breast cancer post-operative metastases reach 30% -40%, of which about 76% die within 2 years after recurrent metastasis. Inhibition of breast cancer invasion and metastasis has become a major challenge to improve survival in breast cancer patients.

Surgical excision, chemotherapy, radiation therapy are conventional treatments. For unresectable tumors, radiation therapy and systemic therapy may be selected. Systemic therapies for lung cancer include conventional chemotherapy, and targeted molecular therapies such as tyrosine kinase inhibitors, inhibitors against EGFR and BRAF. Chemotherapy is beneficial to patients with metastatic disease, and the combination of chemotherapy and radiotherapy is suitable for stage III lung cancer. The use of angiogenesis, epidermal growth factor receptor inhibitors and other novel anti-cancer agents is beneficial for improving patient prognosis levels. Although immune targeted therapies are promising, such as Immune Checkpoint Inhibitors (ICI), large-scale clinical applications have problems with low response, immune-related side effects, economy, etc. Personalized and combination therapies would be future treatments for lung cancer.

2',4' -Trimethoxychalcone (TEC, 2',4' -Trimethoxychalcone), also known as metochalcone, has a molecular formula of C18H18O4, a molecular weight of 298.34, a CAS number of 18493-30-6, and is in the form of pale yellow powder. The metochalcone is an isoliquiritigenin derivative and is obtained by chemical synthesis. The invention provides a synthesis method and a pharmaceutical composition for treating cancers, wherein the pharmaceutical composition comprises metochalcone and a pharmaceutically acceptable carrier or auxiliary material.

However, there are no reports of methods for synthesizing metochalcone and inhibiting cancer.

Disclosure of Invention

In order to solve the problems, the invention provides a new application of the metochalcone in preparing a medicine for inhibiting tumor metastasis.

Use of metochalcone (formula 1) in the preparation of a medicament for inhibiting tumor proliferation and metastasis.

Further, the drug is 2',4' -trimethoxychalcone or (E) -1- (2, 4-dimethoxyphenyl) -3- (4-methoxyphenyl) prop-2-en-1-one.

Further, the breast cancer is lung cancer/breast cancer.

Further, the medicament is capable of inhibiting tumor cell proliferation and migration.

The invention also provides application of the metochalcone in preparing a medicament for preventing and/or treating cancer.

Further, the medicine is a preparation prepared by taking the metochalcone as an active ingredient and adding pharmaceutically acceptable auxiliary materials.

Furthermore, the medicine is a preparation prepared by taking the metochalcone as the only active ingredient and adding pharmaceutically acceptable auxiliary materials.

Further, the preparation is an oral preparation or an injection preparation.

Further, the oral preparation is decoction, oral liquid, granules, capsules, powder, pills or tablets.

The invention has the beneficial effects that:

the invention discovers for the first time that the metochalcone has the capability of obviously inhibiting the proliferation of lung cancer and breast cancer cells, has better tumor inhibition effect in cell experiments, and has good application prospect in preparing medicines for inhibiting tumor metastasis.

It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.

Drawings

FIG. 1 shows the results of inhibition of cell proliferation after treatment of A-549 and BT-549 cells with various concentrations of mitochalcone.

FIG. 2 shows the results of cell migration scratch experiments after treatment of A-549 and BT-549 cells with various concentrations of mitochalcone.

Detailed Description

The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.

The raw materials and equipment used in the invention are all known products and are obtained by purchasing commercial products.

Example 1 Synthesis of Compounds

4-Methoxybenzaldehyde (1) (1.51 g,11.1 mmol) and 2',4' -dimethoxyacetophenone (2) (2.00 g,11.1 mmol) were weighed out into ethanol (100 ml), naOH (977 mg,24.4mmol,2.20 eq) was dissolved into H2O (10 ml) at 0℃and added dropwise to the mixture. The reaction was stirred at 20℃for 24 hours. Hydrochloric acid (2 Mol) was added to adjust the mixture to ph=1 at 0 ℃, and the mixture was filtered, recrystallized, and dried under reduced pressure to obtain the objective compound.

Pale yellow powder in 36.2% yield, 1H NMR (400 mhz, chromaform-d) δ=7.76 (d, j=8.7 hz, 1H), 7.67 (d, j=15.8 hz, 1H), 7.58 (d, j=8.8 hz, 2H), 7.41 (d, j=15.8 hz, 1H), 7.28 (s, 1H), 6.94 (d, j=8.7 hz, 2H), 6.59 (dd, j=2.2, 8.6hz, 1H), 6.53 (d, j=2.2 hz, 1H), MS (ESI) (m/z 299.0[ m+h)] ⁺ 。

Example 2 cell proliferation assay

Cell counting kit 8 (CCK-8) is a widely used cell viability detection method and has the advantages of high sensitivity, low toxicity and convenient operation. The dehydrogenase in the cell mitochondria reduces WST-8 in the kit to generate orange yellow formazan dye, the absorbance is in direct proportion to the cell activity, and the enzyme-labeling instrument measures the absorbance at 450nm to indirectly reflect the cell activity.

1. Experimental materials

Lung cancer a549 cells, breast cancer BT-549 cells, purchased from the national academy of sciences cell bank;

test drug: metochalcone;

culture medium: RPMI 1640+10% fbs+1% diabody;

culture conditions: 37 ℃,5% CO ₂ Culturing in a constant temperature box;

liquid exchange and passage period: the cells were passaged 1-2 days after 1-fold change until 80% confluence was achieved.

2. Experimental grouping

(1) Control group: BT-549 cells 0 μm metochalcone treatment group;

(2) Experimental group: BT-549

cells

5, 10, 20, 40, 50 μm metochalcone treatment;

each group of 3 samples.

3. Experimental method

(1) Cells in the logarithmic growth phase are taken, separated by a pancreatin method and counted by using a cell counting plate, resuspended in a medium containing 10% FBS and prepared into a single cell suspension containing 3.5X10-4/ml for later use.

(2) PBS was added to the 96-well plate at four weeks, cell suspension plates were added in the middle, and the plating cell density was 5000 cells/well. The last 2 columns were supplemented with serum-free medium alone as a control. After incubation overnight in a 5% CO2, 37℃incubator, the culture broth was discarded after cell attachment was observed.

(3) TEC (50 mM stock solution prepared by dissolving DMSO) was added to each column in a concentration gradient, and the concentration was measured at (0, 5, 10, 20, 40, 50. Mu.M).

(4) After 24h and 48h, 100ul of culture medium containing 1% serum was added to each well, and incubation was continued with 10ul of CCK-8 reagent (protected from light) at 37℃for 1 hour, and OD was measured at 450nm using an ELISA reader. P <0.05, P <0.01, P <0.001 compared to the control group (0 μm).

The cell proliferation inhibition was calculated according to the following formula:

(1- (O.D. dosing/O.D. blank)) ×100%

4. Experimental results

The results of inhibition of cell proliferation after treatment of A-549 and BT-549 cells with various concentrations of mitochalcone are shown in FIG. 1.

The results show that lung cancer a549 cells and breast cancer cells BT-549 cells proliferated significantly less (P < 0.001) after administration of metoprolol compared to the control group without metoprolol. Half-maximal inhibitory concentrations were 22.05 and 22.67 μm after 24 hours of drug treatment, respectively, and 4.28 and 3.4 μm after 48 hours of dosing. It is demonstrated that metochalcone can significantly inhibit the proliferation of lung cancer cells and breast cancer cells, and the inhibition increases with increasing drug concentration.

Example 3 cell migration experiment

Experimental materials and groupings were as in example 2. Cells growing well in log phase were digested with pancreatin and cell density was adjusted to about 2.5X10 with complete medium of 10% serum ⁵ 2mL of the cell suspension was pipetted into a six well plate.

(2) After incubation for 24 hours, scratches were made in six well plates with a 10. Mu.L tip, washed once with PBS, 1% serum-containing solution was added to each group, and photographed under a 100X microscope.

(3) After 24 hours incubation, the cells were washed once with PBS and photographed again.

(4) Counting the width change of each group of scratches to evaluate the migration condition of the cells; p <0.05, P <0.01, P <0.001, compared to the control group.

The results of cell migration scratch experiments after treatment of A-549 and BT-549 cells with various concentrations of mitochalcone are shown in FIG. 2.

The change in scratch width can directly reflect the migration ability of tumor cells. Experimental results show that lung cancer a-549 cells and breast cancer cells BT-549 groups showed a reduced scratch width (P < 0.05) after administration of metoprolol compared to the control group without metoprolol, indicating that metoprolol can significantly migrate to triple negative breast cancer cells, and that inhibition increases with increasing drug concentration.

In summary, the invention provides the synthesis of metochalcone and the use thereof in the preparation of a medicament for inhibiting tumor metastasis. The invention discovers for the first time that the metochalcone is convenient to synthesize, has the capability of obviously inhibiting proliferation of the triple-negative breast cancer cells, can also obviously inhibit migration of the triple-negative breast cancer cells, and can effectively inhibit metastasis of the triple-negative breast cancer. The metochalcone has good application prospect in preparing medicines for inhibiting tumor metastasis.

Claims

1. Use of metochalcone (formula 1) in the preparation of a medicament for inhibiting tumor proliferation and metastasis.

2. Use according to claim 1, characterized in that: the tumor is lung cancer and breast cancer.

3. Use according to any one of claims 1-2, characterized in that: the medicine can inhibit proliferation and migration of tumor cells.

4. Use of metochalcone in the manufacture of a medicament for the prevention and/or treatment of lung cancer and/or breast cancer.

5. Use according to any one of claims 1 to 4, characterized in that: the medicine is a preparation prepared by taking the metochalcone as an active ingredient and adding pharmaceutically acceptable auxiliary materials.

6. Use according to claim 5, characterized in that: the medicine is a preparation prepared by taking the metochalcone as the only active ingredient and adding pharmaceutically acceptable auxiliary materials.

7. Use according to claim 6, characterized in that: the preparation is oral preparation or injection preparation.

8. Use according to claim 7, characterized in that: the oral preparation is decoction, oral liquid, granule, capsule, powder, pill or tablet.