Compound as well as preparation method and application thereof

Abstract

The invention relates to the technical field of biological medicines, and particularly discloses a compound as well as a preparation method and application thereof. The compound has a structure as shown in a general formula (I). Researches show that the compound has a remarkable inhibition effect on the growth of breast cancer and lung cancer cells; further researches show that the compound can also significantly inhibit migration and invasion of breast cancer and lung cancer cells. Therefore, the compound disclosed by the invention is used as an effective component for preparing the medicine with the effects of resisting breast cancer and lung cancer; particularly, the compound has important application value when being used for preparing medicines with effects of resisting metastasis and invasion of breast cancer and lung cancer cells.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a compound, a preparation method and an application thereof. Background Art

[0002] Globally, new cases and cancer deaths continue to rise, and malignant tumors have become a key cause of death in China. Approximately 90% of cancer deaths are due to distant metastasis. Once cancer cells metastasize, the disease rapidly worsens, significantly increasing the difficulty of treatment. However, the current lack of effective anti-metastasis drugs in clinical practice presents a significant challenge in cancer treatment. Against this backdrop, the "Mechanisms of Tumor Metastasis and Development of New Anti-metastasis Drugs" was listed as one of my country's 60 major scientific and engineering challenges by the China Association for Science and Technology in 2018, a topic of immense significance. Only by deeply understanding the mechanisms of tumor metastasis and successfully developing effective new anti-metastasis drugs, combined with existing anti-cancer proliferative drugs, can cure rates be improved, survival rates prolonged, and hope brought to countless cancer patients.

[0003] Therefore, providing a compound with anti-cancer effect, especially anti-cancer cell metastasis effect, has important application value for the treatment of cancer. Summary of the Invention

[0004] In order to overcome at least one of the technical problems existing in the prior art, the present invention provides a compound and a preparation method and application thereof.

[0005] The technical solutions of the present invention are as follows: The present invention first provides a compound having a structure shown in general formula (I):

[0006] Wherein, R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12, R13, R14, and R15 are selected from any one of fluorine, bromine, iodine, a hydrogen atom, or any substituent, and may be monosubstituted or polysubstituted.

[0007] Preferably, the compound has the following structure: .

[0008] The compound of the above specific structure is abbreviated as EH-M005; its specific name is: 7-(4-(methylsulfonyl)benzamido)-N-phenyl-2-naphthamide.

[0009] The present invention also provides a method for preparing the above compound, which uses 7-(4-(methylsulfonyl)benzamido)-2-naphthoic acid and aniline as raw materials.

[0010] Preferably, the preparation method of the compound comprises the following steps: dissolving 7-(4-(methylsulfonyl)benzamido)-2-naphthoic acid in an organic solvent, then adding a condensing agent and aniline, and stirring the mixture at room temperature for 12 to 24 hours; after the reaction, separating the product to obtain 7-(4-(methylsulfonyl)benzamido)-N-phenyl-2-naphthoamide, i.e., the compound of the above-mentioned specific structure (EH-M005).

[0011] Preferably, the molar ratio of 7-(4-(methylsulfonyl)benzamido)-2-naphthoic acid to aniline is 1:1-2.

[0012] Most preferably, the molar ratio of 7-(4-(methylsulfonyl)benzamido)-2-naphthoic acid to aniline is 1:1.5.

[0013] Preferably, the ratio of 7-(4-(methylsulfonyl)benzamido)-2-naphthoic acid to the organic solvent is 100 mg:1-3 mL.

[0014] Preferably, the organic solvent is N,N-dimethylformamide.

[0015] Preferably, the condensing agent is selected from a combination of 1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.

[0016] The present invention also provides an application of the compound in preparing a drug with anti-cancer effect.

[0017] Preferably, the anti-cancer agent is anti-breast cancer.

[0018] Preferably, the anti-cancer agent is anti-lung cancer.

[0019] The present invention also provides a use of the compound in preparing a drug capable of inhibiting cancer cell metastasis or cancer cell invasion.

[0020] Preferably, the cancer cells are breast cancer cells.

[0021] Preferably, the cancer cells are lung cancer cells.

[0022] Beneficial Effects: The present invention provides a novel compound; studies have shown that the compound significantly inhibits the growth of breast cancer and lung cancer cells; further studies have shown that the compound can also significantly inhibit the migration and invasion of breast cancer and lung cancer cells. Therefore, the compound of the present invention has important application value when used as an active ingredient in the preparation of drugs with anti-breast cancer and anti-lung cancer effects, especially in the preparation of drugs with anti-metastasis and anti-invasive effects of breast cancer and lung cancer cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The synthetic route diagram of the compound of the present invention.

[0024] Figure 2 This is the hydrogen spectrum of EH-M005.

[0025] Figure 3 The figure shows the IC50 experimental results of EH-M005 on breast cancer cells (MDA-MB-231) and lung cancer cells (H1299) after 48 hours of MTT detection.

[0026] Figure 4 The figure shows the experimental results showing that EH-M005 drug significantly inhibits the migration of breast cancer cells (MDA-MB-231) at concentrations of 0.5 μM and 5 μM.

[0027] Figure 5 This figure shows the experimental results showing that EH-M005 drug significantly inhibits the migration of lung cancer cells (H1299) at concentrations of 5 μM and 10 μM.

[0028] Figure 6 The graph shows the results of an invasion experiment in which EH-M005 inhibited the breast cancer cell line MDA-MB-231 at concentrations of 0.5 μM, 2 μM, and 5 μM.

[0029] Figure 7 The graph shows the results of an experiment in which EH-M005 inhibits the invasion of lung cancer cells H1299 at concentrations of 2 μM, 10 μM, and 20 μM. DETAILED DESCRIPTION

[0030] The present invention will be further described in detail below with reference to specific examples, but the examples do not limit the present invention in any form.

[0031] Example 1 Preparation of Compounds To a solution of 7-(4-(methylsulfonyl)benzamido)-2-naphthoic acid (200 mg, 0.54 mmol) in N,N-dimethylformamide (DMF, 2 ml) were added 1-hydroxybenzotriazole (HOBt, 73 mg, 0.54 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI, 156 mg, 0.81 mmol) and aniline (61 mg, 0.65 mmol), and the mixture was stirred at room temperature for 16 h. To the mixture was added 2 mL of DMF for dilution, and then purified by preparative high performance liquid chromatography (mobile phase: water / acetonitrile containing 0.05% formic acid) to obtain 7-(4-(methylsulfonyl)benzamido)-N-phenyl-2-naphthamide, compound EH-M005 (160 mg, yield: 66.48%), as an off-white solid.

[0032] The mass spectrum and hydrogen spectrum data of 7-(4-(methylsulfonyl)benzamido)-N-phenyl-2-naphthamide (abbreviated as EH-M005) are as follows: electrospray ionization mass spectrum (ESI-MS): (M-H)⁻ = 443.1; 1 H NMR (400 MHz, DMSO- d 6) δ 10.81 (s, 1H), 10.41 (s, 1H), 8.67 (s, 1H), 8.56 (s,1H), 8.25 (d, J = 8.4 Hz, 2H), 8.13 (d, J = 8.4 Hz, 2H), 8.03 (dd, J = 8.8, 5.6 Hz,2H), 7.97 – 7.89 (m, 2H), 7.84 (d, J = 8.0 Hz, 2H), 7.39 (t, J = 8.0 Hz, 2H),7.13 (t, J = 7.6 Hz, 1H), 3.32 (s, 3H). Experimental Example 1 MTT assay for the effect of drugs on cell growth 1.1 Experimental subjects: breast cancer cells MDA-MB-231; lung cancer cells H1299; 1.2 Experimental drug: the compound prepared in Example 1, namely compound EH-M005, and the compound solvent DMSO.

[0033] 1.3 Experimental Methods: On the afternoon of the first day, cells were plated: logarithmic-phase cells were harvested, the cell suspension concentration was adjusted, and 5,000 cells / 90 μl of cell suspension were added to each well. The next morning, drug was added: 10 μl of a gradient of drug concentrations was added to each well, with triplicate wells for each drug concentration. The cells were incubated in a 5% CO2, 37°C incubator. 48 hours after drug addition, the plates were harvested: First, the cells were visually inspected under an inverted microscope. Then, 20 μl of MTT solution was added to each well. After incubation at 37°C for 4 hours, the medium was aspirated, and 100 μl of DMSO was added to each well. Crystals were dissolved by shaking. OD values ​​were measured: The absorbance of each well was measured at 490 nm on a microplate reader. Relative cell viability or drug inhibition was calculated. A blank control group was established for this experiment. A DMSO solution containing the compound at the same dilution ratio was added to each well as a negative control group. The experimental group contained compound EH-M005.

[0034] 1.4 Calculation: Relative survival rate = (OD value of compound experimental group - OD value of blank control group) / (OD value of negative control group - OD value of blank control group) × 100%; Drug inhibition rate = 1 - relative survival rate.

[0035] The results show that: Figure 3 It can be seen that the IC values ​​of compound EH-M005 for breast cancer cells MDA-MB-231 and lung cancer cells H1299 are within the soluble concentration range. 50 The concentrations of the two compounds were all greater than 200 μM, and the cytotoxicity was very low.

[0036] Experimental Example 2: Cell scratch assay to determine the effect of drug EH-M005 on cell motility / migration 2.1 Experimental subjects: Human breast cancer cell line MDA-MB-231; human lung cancer cell line H1299.

[0037] 2.2 Experimental drug: the compound prepared in Example 1, namely compound EH-M005, and the compound solvent DMSO.

[0038] 2.3 Experimental Method: Before plating, use a marker pen with a ruler to evenly draw horizontal lines across the wells, approximately every 0.5-1 cm. Three lines are drawn, designated as lines a, b, and c. Line b runs through the center, and the other two lines are drawn equally spaced on either side of line b. For plating, add 2 ml of complete medium containing 10% FBS to each well of the 6-well plate. 2 × 10 6Set up two replicate wells for each well and culture for approximately 20-24 hours. The optimal cell number should be at least 90% confluence after overnight attachment, and this number can be adjusted as needed. The next day, open the plate lid, aspirate the remaining medium, and place a ruler perpendicular to the line b on the plate. Using a 200 μl pipette tip, move the ruler up and down evenly to create a streak line. Similarly, make two parallel lines equally spaced on either side of this line, designated Line 1, Line 2, and Line 3 from left to right. Rinse the cells three times with sterile 1× PBS. After removing the streaked cells, add complete medium supplemented with 20% FBS and compound EH-M005 (working concentrations: 0.5 μM and 5 μM for breast cancer cells MDA-MB-331; 5 μM and 10 μM for lung cancer cells H1299). Incubate at 37°C in a 5% CO2 incubator. Samples were taken and photographed 0 and 24 hours after drug addition.

[0039] The results show that: Figure 4 and 5 It can be seen that after 30 h of action, compound EH-M005 can significantly inhibit the migration of human breast cancer cells MDA-MB-231 at concentrations of 0.5 μM and 5 μM, and inhibit the migration of human lung cancer cells H1299 at concentrations of 5 μM and 10 μM.

[0040] Experimental Example 3: Cell invasion assay to determine the effect of drugs on cell invasion ability 3.1 Experimental subjects: Breast cancer cells 231; lung cancer cells H1299.

[0041] 3.2 Experimental drug: the compound prepared in Example 1, namely compound EH-M005, and the compound solvent DMSO.

[0042] 3.3 Experimental Methods: Preparation of cell suspension: Digest the cells, centrifuge and discard the culture medium after termination of digestion, wash once with PBS, resuspend in serum-free medium, and adjust the cell density to the appropriate concentration (the seeding density of breast cancer 231 cells is 5×10 4 The seeding density of lung cancer cells H1299 was 4.5×10 4 The seeding density of gastric cancer cell MGC803 was 8×10 4 The seeding density of pancreatic cancer cells PANC-1 was 6×10 4Cells were seeded at a concentration of 100 μL / 200 μL. For each cell type, a negative control group (using DMEM at the same dilution ratio as compound EH-M005) and a treatment group (compound EH-M005 at working concentrations of 0.5, 2, and 5 μM for MDA-MB-231 cells and 2, 10, and 20 μM for lung cancer H1299 cells) were set up in triplicate. Based on the cell density, an appropriate amount of cell suspension was taken, and an appropriate volume of 10% BSA was added to a final concentration of 0.1%. Compound EH-M005 was then added, and the volume was filled with DMEM to a total of 200 μL per well. After mixing, the cells were gently and evenly added to the upper chamber of the Transwell plate. Immediately, 800 μL of culture medium containing 20% ​​FBS and the corresponding working concentration of drug was added to the lower chamber of the 24-well plate. The plates were harvested approximately 24 hours after treatment. For cell staining, the Transwell chamber was removed, the culture medium in the wells was discarded, and the cells were fixed with methanol for 30 minutes. The methanol was then aspirated. Stain with 0.1% crystal violet for 30 minutes, aspirate to recover the crystal violet, and wash once with PBS. Gently wipe away any unmigrated cells in the upper chamber with a cotton swab, wash twice with PBS, and place in a clean 24-well plate. Place the chamber in a fume hood to air dry. Results: Observe under a 5X microscope and observe cells in five random fields under a 10X microscope. Photograph, count, and plot.

[0043] The results show that: Figure 6 and 7 It can be seen that compound EH-M005 can significantly inhibit the invasion of breast cancer cells MDA-MB-231 at 0.5, 2, and 5 μM; and can significantly inhibit the invasion of lung cancer cells H1299 at a concentration of 20 μM.

Claims

1. A compound, characterized in that The compound has a structure shown in general formula (I): ； Wherein, R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12, R13, R14, and R15 are selected from any one of fluorine, bromine, iodine, a hydrogen atom, or any substituent, and may be monosubstituted or polysubstituted.

2. The compound according to claim 1, characterized in that The compound has the following structure: 。 3. The method for preparing the compound according to claim 2, characterized in that: It is prepared from 7-(4-(methylsulfonyl)benzamido)-2-naphthoic acid and aniline.

4. The method for preparing the compound according to claim 2, characterized in that: The method specifically comprises the following steps: dissolving 7-(4-(methylsulfonyl)benzamido)-2-naphthoic acid in an organic solvent, then adding a condensing agent and aniline, and stirring the mixture at room temperature for 12 to 24 hours; after the reaction, separating the product to obtain 7-(4-(methylsulfonyl)benzamido)-N-phenyl-2-naphthoamide, which is the compound of the structure described in claim 2.

5. The method for preparing the compound according to claim 4, characterized in that: The molar ratio of 7-(4-(methylsulfonyl)benzamido)-2-naphthoic acid and aniline is 1:1~2; Most preferably, the molar ratio of 7-(4-(methylsulfonyl)benzamido)-2-naphthoic acid to aniline is 1:1.

5.

6. The method for preparing the compound according to claim 4, characterized in that: The ratio of 7-(4-(methylsulfonyl)benzamido)-2-naphthoic acid to organic solvent is 100 mg:1~3 mL.

7. The method for preparing the compound according to claim 4, characterized in that: The organic solvent is N,N-dimethylformamide.

8. The preparation method according to claim 3, characterized in that The condensing agent is selected from a combination of 1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.

9. Use of the compound according to claim 1 or 2 in the preparation of a drug having an anti-cancer effect.

10. Use of the compound according to claim 1 or 2 in the preparation of a drug having the effect of inhibiting cancer cell metastasis or cancer cell invasion.

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