阅读说明：本技术 一种具有抑制乳腺癌细胞增殖、转移活性的化合物及在制备抗乳腺癌的药物中的应用 (Compound with breast cancer cell proliferation and metastasis inhibiting activity and application thereof in preparation of anti-breast cancer drugs ) 是由 郭守山 郭守河 于 2020-12-17 设计创作，主要内容包括：本发明公开了一种具有抑制乳腺癌细胞增殖、转移活性的化合物及在制备抗乳腺癌的药物中的应用。本发明发现如下结构的吲哚衍生物一方面可以抑制乳腺癌细胞的增殖,另一方面可以抑制乳腺癌细胞的侵袭转移,具有开发成治疗乳腺癌的药物的前景：(The invention discloses a compound with the activities of inhibiting proliferation and metastasis of breast cancer cells and application thereof in preparing a medicament for resisting breast cancer. The invention discovers that the indole derivative with the following structure can inhibit the proliferation of breast cancer cells on one hand and inhibit the invasion and metastasis of the breast cancer cells on the other hand, and has the prospect of developing a medicament for treating breast cancer:)

1. The medical application of the indole derivative with the following chemical structure in preparing the medicine for treating breast cancer.

2. The use of the indole derivative of claim 1 for the preparation of a medicament for inhibiting breast cancer cell proliferation.

3. The use of the indole derivative of claim 1 for the preparation of a medicament for inhibiting invasion and metastasis of breast cancer cells.

4. A medical use according to claims 1 to 3, wherein: the medicament is in the form of a pharmaceutical preparation, and the active ingredient is the indole derivative.

5. The medical use according to claim 4, wherein: the pharmaceutical preparation can be in the form of tablets or capsules or injections and the like.

Technical Field

The invention belongs to the field of medicines, and particularly relates to a compound with activities of inhibiting proliferation and metastasis of breast cancer cells and application of the compound in preparation of a medicine for resisting breast cancer.

Background

The breast cancer is a common malignant tumor in China, the incidence rate is gradually increased and the breast cancer is younger, and the health of women is seriously threatened. In the treatment of tumors and cancers, if powerful attacks can be given to the specific molecular changes of the cancer, the treatment effect will be greatly improved. The small molecular medicine can specifically block the signal conduction path necessary in the growth and proliferation process of tumor, so as to achieve the purpose of treatment. Compared with macromolecular drugs, the measuring method of the small-molecule chemical drug is relatively simple, and the discovery and the modification of the small-molecule chemical drug are relatively easy. In recent years, the small molecule drug targeted therapy of breast cancer has achieved remarkable curative effect in clinical practice, and the therapy of breast cancer is pushed to an unprecedented new stage (Linzhonlin, etc., research progress of small molecule drugs for treating breast cancer, Chinese clinical pharmacology and therapeutics, 2015).

Indole compounds are important compounds, and different indole derivatives have different pharmacological activities. For example, the indole derivatives 1-3 having the following structures are found by the applicant in the early stage that both indole derivatives 1 and 2 have significant proliferation inhibitory activity on acute lymphoblastic leukemia Jurkat cells, while indole derivative 3 has insignificant inhibitory activity (application No. 2020114129649).

Further research shows that the indole derivative 3 has other biological activity, and the invention is provided.

Disclosure of Invention

The invention aims to provide a compound with activities of inhibiting proliferation and metastasis of breast cancer cells and application of the compound in preparation of a medicine for resisting breast cancer.

The technical scheme for realizing the aim of the invention is as follows:

the medical application of the indole derivative with the following chemical structure in preparing the medicine for treating breast cancer.

The indole derivative is used for preparing a medicine for inhibiting breast cancer cell proliferation.

The indole derivative is used for preparing a medicine for inhibiting invasion and metastasis of breast cancer cells.

Further, the medicament is in the form of a pharmaceutical preparation, and the active ingredient is the indole derivative.

Further, the pharmaceutical preparation form can be tablets or capsules or injection and the like.

The invention has the outstanding advantages that:

the invention discovers that the indole derivative with the following structure can inhibit the proliferation of breast cancer cells on one hand and inhibit the invasion and metastasis of the breast cancer cells on the other hand, and has the prospect of developing a medicament for treating breast cancer.

Drawings

FIG. 1 shows the chemical structures of indole derivatives 1, 3 to be tested;

FIG. 2 shows the inhibition rate of MCF-7 cell proliferation by different concentrations of drug.

Detailed Description

The following detailed description of the present invention is provided in connection with the examples, and for reasons of brevity, the description of the experimental procedures is not intended to be exhaustive, and all parts not specifically described in the experiments are routine procedures well known to those skilled in the art.

Example 1

First, experimental material

Human breast cancer MCF-7 cells were purchased from ATCC and cultured in RPMI-1640 containing 10% fetal bovine serum at 37 ℃ in 5% CO₂And (5) performing conventional culture in a cell culture box.

RPMI-1640 medium and fetal bovine serum were purchased from Gibco, USA.

The indole derivatives 1, 3 to be tested shown in FIG. 1 have a purity of not less than 98%.

Second, Experimental methods

1. Cell culture

Human breast cancer MCF-7 cells are cultured in RPMI-1640 culture medium containing 10% fetal calf serum and 1% double antibody at 37 deg.C with volume fraction of 5% CO₂And culturing in a saturated humidity incubator by conventional method, and replacing the culture solution every 2d for 1 time.

2. MTT method for measuring cell proliferation activity

Digesting and re-suspending human breast cancer MCF-7 cells in logarithmic growth phase to prepare cell suspension, and inoculating the cell suspension to 96 holesIn the plate, each hole is 1X 10⁴Cell, 200. mu.L, 37 ℃ and 5% CO₂Culturing for 24h under the condition, replacing with RPMI-1640 medium containing different concentrations (1, 2.5, 5, 10, 20 μ M) of indole derivatives 1 or 3 and 10% of fetal bovine serum and 1% of double antibody, and culturing for 48h while setting blank control groups without drug, each 5 duplicate wells. And after the time is up, adding 20 mu L of MTT reagent with the concentration of 5mg/mL into each hole, continuously culturing for 4h, removing the supernatant, adding 150 mu L of LDMSO into each hole, oscillating and dissolving, measuring the absorbance value (OD570) at the wavelength of 570nm by using an enzyme-labeling instrument, and calculating the proliferation inhibition rate of the indole derivatives 1 or 3 with different concentrations on MCF-7 cells according to a formula.

3. Transwell method for measuring cell invasion transfer activity

Add 50. mu.L of matrix gel to the Transwell chamber and coagulate it at 37 ℃ and wash the upper chamber with serum-free RPMI-1640 medium for use. Taking MCF-7 cells in logarithmic growth phase, preparing into cell density of 5 × 10 with serum-free RPMI-1640 culture solution⁵one/mL cell suspension, inoculated in the upper chamber, each chamber of 200 u L, and added indole derivatives 1 or 3 to make its final concentration of 0, 1, 2.5M; 200 μ L of RPMI-1640 medium containing 10% fetal bovine serum was added to each corresponding lower chamber. At 37 deg.C and 5% CO by volume₂And after the cells are cultured for 48 hours in an incubator with saturated humidity, sucking the cell suspension in the upper chamber, washing off the cells which do not pass through the matrigel, fixing the cells by using 4% paraformaldehyde and staining the cells by using 1% crystal violet, randomly selecting 3 visual fields under an optical microscope, and calculating the number of transmembrane cells.

4. Statistical method

All data were statistically analyzed using SPSS17.0, the data were expressed as mean. + -. standard deviation, the comparison between the two groups was performed using t-test, and the difference P < 0.05 was statistically significant.

Third, experimental results

1. Cell proliferation Activity

The MTT method experiment results are shown in Table 1 and figure 2, the indole derivative 3 with the concentration of 5-20 mu M has obvious proliferation inhibition activity on human breast cancer MCF-7 cells, and the indole derivative 1 with the test concentration has no obvious proliferation inhibition activity on the human breast cancer MCF-7 cells. The experimental result shows that the indole derivative 3 has the drug effect of resisting human breast cancer MCF-7 cells.

TABLE 1 inhibition of MCF-7 cell proliferation (%). by different drug concentrations

Drug concentration	Indole derivatives 3	Indole derivatives 1
			1μM	2.15±1.32	2.32±1.15
2.5μM	2.29±1.27	2.26±1.30
			5μM	9.64±2.89	2.58±1.24
10μM	20.52±2.75	2.94±1.62
			20μM	38.95±3.12	2.88±1.57

2. Cell invasion transfer Activity

Selecting the drug concentration of 1 and 2.5 mu M which has no obvious proliferation inhibition activity on human breast cancer MCF-7 cells to carry out invasion transfer activity test. The results of the Transwell method are shown in table 2. Compared with the control group containing 0 μ M of the drug, the indole derivative 3 group had significantly reduced number of transmembrane cells and had a concentration-dependent effect. The number of membrane-penetrating cells in the indole derivative 1 group was not significantly changed from that in the control group containing 0. mu.M of the drug.

TABLE 2 number of transmembrane cells in each group

Drug concentration	Indole derivatives 3	Indole derivatives 1	Control group
				0μM	/	/	155.67±5.03
1μM	91.67±4.51	153.33±5.51	/
				2.5μM	54.00±6.56	155.50±4.95	/

Example 2

A tablet, capsule or injection for inhibiting proliferation or/and invasion and metastasis of breast cancer cells comprises indole derivatives shown in formula 3 in figure 1 as active ingredient, and common adjuvants for tablet, capsule or injection.

The foregoing embodiments are provided to illustrate the present invention more fully, but those skilled in the art will appreciate that the scope of the present invention should not be limited to the specific embodiments described above.