Demethylzeylasteral Schiff base derivative and application thereof in preparation of anti-cancer drugs

Abstract

The invention discloses a demethylzeylasteral Schiff base derivative and application thereof in preparation of anti-cancer drugs, the demethylzeylasteral Schiff base derivative has a structural formula as shown in a formula 1, a synthetic route is as shown in a formula 2, and the demethylzeylasteral Schiff base derivative has obvious anti-tumor cell activity; the invention also provides an application of the demethylzeylasteral Schiff base derivative in preparation of anti-cancer drugs, especially in preparation of drugs for resisting colon cancer, ovarian cancer and liver cancer. The invention also provides an application of the demethylzeylasteral Schiff base derivative or the medicinal salt thereof in preparation of HCT116, SKOV3 and HepG2 cell proliferation inhibitors. In-vitro experiments prove that the demethylzeylasteral Schiff base derivative has an obvious inhibiting effect on cell growth and cell viability of HCT116, SKOV3 and HepG2, not only has the potential of being developed into a new generation of anti-cancer drugs, but also has potential application value in the fields of cancer targeted drugs, probe design and fluorescent biomarkers.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicine, and particularly relates to norzelumbal schiff base derivatives and their application in the preparation of anti-cancer drugs. Background Art

[0002] Malignant tumors are a class of diseases that threaten human health and life. Currently, the treatment of malignant tumors often adopts a comprehensive measure combining surgery, radiotherapy, and chemotherapy. Chemotherapy is a systemic treatment and can eliminate cancer cells with distant metastasis, playing an important role in the comprehensive treatment. However, there are problems with the efficacy, side effects of chemotherapy drugs, and drug resistance of tumor cells in clinical practice. Therefore, it is still an urgent task to search for highly effective and low-toxic anti-tumor drugs from different perspectives.

[0003] Natural products are an important source of lead compounds for drugs and functional organic molecules. Screening lead compounds for drugs from natural products has always been an effective way for new drug research and development. Due to their extensive biological activities and low toxic side effects, natural products have always been an important source for drug research and development. Therefore, the active ingredients in traditional Chinese medicine have become a current research hotspot. Among them, norzelumbal is isolated from the roots of Tripterygium wilfordii and has various biological activities, including anti-tumor, anti-inflammatory, immunosuppressive effects, etc. Among them, the anti-tumor activity of norzelumbal has been widely studied, and it shows cytotoxicity to a variety of human cancer cell lines by inducing apoptosis. However, the anti-tumor efficacy (HCT116: 26.19 ± 2.43 μM, SKOV3: 11.76 ± 0.30 μM, HepG2: 26.54 ± 2.89 μM) and physicochemical properties of norzelumbal still need to be improved. By modifying the C-4 position of norzelumbal and designing different substituents, and using the CCK8 method to measure the inhibitory effects of the compounds on three cancer cells, HCT116, SKOV3, and HepG2, it can provide a good starting point for the development of new anti-tumor drugs. Summary of the Invention

[0004] In view of this, the main purpose of the present invention is to provide a norzelumbal schiff base derivative, which has an obvious inhibitory effect on cancer cells.

[0005] Another purpose of the present invention is to provide the application of the above-mentioned norzelumbal schiff base derivative in the preparation of anti-cancer drugs. The norzelumbal schiff base derivative has obvious anti-tumor cell activity and is used to develop a new type of anti-cancer drug.

[0006] The above objects of the present invention are achieved by the following technical solutions:

[0007] The first aspect of the present invention provides a norzelumbal schiff base derivative, which has a structural formula shown in Formula 1:

[0008]

[0009] In Formula 1, R is selected from one of the following substituents:

[0010] The second aspect of the present invention provides a preparation method of the above-mentioned norzelalamide Schiff base derivatives, and the synthetic route is as

[0011] shown in Formula 2:

[0012]

[0013] The third aspect of the present invention provides the use of the above-mentioned norzelalamide Schiff base derivatives or their pharmaceutically acceptable salts in the preparation of anticancer drugs.

[0014] Preferably, the cancers include colon cancer, ovarian cancer and liver cancer, specifically colon cancer HCT116, ovarian cancer SKOV3, and liver cancer HepG2.

[0015] The fourth aspect of the present invention provides an anticancer pharmaceutical composition, which includes norzelalamide Schiff base derivatives or their pharmaceutically acceptable solvates, and uses them as active ingredients. The structural formula of the norzelalamide Schiff base derivatives is as described above.

[0016] Preferably, in the pharmaceutical composition, the weight percentage content of the norzelalamide Schiff base derivatives or their pharmaceutically acceptable solvates is 5% to 100%.

[0017] Preferably, the pharmaceutical composition further includes a pharmaceutically acceptable carrier or diluent.

[0018] "Pharmaceutically acceptable carrier" refers to the inactive ingredients in the pharmaceutical composition, including but not limited to calcium carbonate, calcium phosphate, various sugars such as lactose, mannitol, etc., starch, cyclodextrin, magnesium stearate, cellulose, magnesium carbonate, acrylic polymers or methacrylic polymers, gels, water, polyethylene glycol, propylene glycol, ethylene glycol, castor oil or hydrogenated castor oil or polyethoxylated hydrogenated castor oil, sesame oil, corn oil, peanut oil.

[0019] "Pharmaceutically acceptable diluent" includes but not limited to starch (such as corn starch, wheat starch, potato starch, etc.), lactose, dextrin, sucrose, pregelatinized starch, microcrystalline cellulose, inorganic salts (such as calcium hydrogen phosphate, calcium sulfate, calcium carbonate, etc.) and mannitol.

[0020] Preferably, the dosage form of the pharmaceutical composition is selected from at least one of tablet, capsule, granule, dripping pill, suspension, syrup, enteric preparation, emulsion suspension and injection.

[0021] The fifth aspect of the present invention provides the use of the norzelalamide Schiff base derivatives or their pharmaceutically acceptable salts in the preparation of HCT116, SKOV3, and HepG2 cell proliferation inhibitors.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] (1) The present invention confirms through in vitro experiments that the norzelalamide Schiff base derivatives have obvious inhibitory effects on the growth and cell viability of HCT116, SKOV3, and HepG2 cells, providing candidate lead compounds for the research and development of anti-cancer drugs.

[0024] (2) The norzelalamide Schiff base derivatives in the present invention not only have the potential to be developed into a new generation of anti-cancer drugs, but also have potential application values in the fields of cancer targeted drugs, probe design, and fluorescent biological labeling. Description of the Drawings

[0025] Figure 1 Results of the inhibitory effects of different concentrations of Compound 1 in Example 3 and its combination with paclitaxel on the cell cycle of HCT116 cells.

[0026] Figure 2 Results of the inhibitory effects of different concentrations of Compound 2 in Example 3 and its combination with paclitaxel on the cell cycle of HCT116 cells.

[0027] Figure 3 Results of the inhibitory effects of different concentrations of Compound 3 in Example 3 and its combination with paclitaxel on the cell cycle of HCT116 cells.

[0028] Figure 4 Results of the inhibitory effects of different concentrations of Compound 1 in Example 4 and its combination with paclitaxel on the apoptosis of HCT116 cells.

[0029] Figure 5 Results of the inhibitory effects of different concentrations of Compound 2 in Example 4 and its combination with paclitaxel on the apoptosis of HCT116 cells.

[0030] Figure 6 Results of the inhibitory effects of different concentrations of Compound 3 in Example 4 and its combination with paclitaxel on the apoptosis of HCT116 cells. Detailed Embodiments

[0031] To more fully understand the technical solutions, objectives, and advantages of the present invention, the technical effects produced by the present invention will be further described in detail and completely below in conjunction with the accompanying drawings and specific embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of them. It should be noted that for those of ordinary skill in the art, other embodiments obtained without departing from the concept of the present invention all fall within the protection scope of the present invention.

[0032] Unless otherwise specified in the following embodiments, the reagents and materials used are commercially available.

[0033] In the following embodiments, the norzelalamide Schiff base derivatives have the structural formula shown in Formula 1:

[0034]

[0035] In Formula 1, R is selected from one of the following substituents:

[0036] In the following embodiments, there are no special requirements for the preparation method of the above-mentioned norzelalamide Schiff base derivatives, and the method well-known to those skilled in the art can be used for preparation. The synthesis route is shown in Formula 2:

[0037]

[0038] Example 1

[0039] (1) Preparation of Compounds 1-3: Dissolve norzelalamide (50 mg, 0.1 mmol) in anhydrous ethanol (1 mL), and add aniline (9.5 μL, 0.1 mmol), 4-(2-furyl)aniline (16.6 mg, 0.1 mmol), and 4-(2-thienyl)aniline (18.2 mg, 0.1 mmol) respectively, and react at room temperature for 5-6 h.

[0040] (2) Filter the reaction mixture, and evaporate the filtrate under reduced pressure using a rotary evaporator. Finally, purify by gel column chromatography to obtain pure target derivatives 1-3 (Compound 1: 48.7 mg, yield 84.0%; Compound 2: 61.0 mg, yield 94.3%; Compound 3: 52.0 mg, yield 78.4%).

[0041] Compound 1 1 H NMR(500MHz,CDCl 3)δ: 10.49 (s, 1H), 7.44 (s, 4H), 7.13 (s, 1H), 6.68 (d, J = 10 Hz, 1H), 6.23 (s, 1H), 2.40 (s, 1H), 2.15 (s, 1H), 2.06 (s, 1H), 1.80 (s, 2H), 1.63 (s, 2H), 1.55 (s, 2H), 1.48 (s, 3H), 1.35 (s, 1H), 1.30 (s, 1H), 1.24 (s, 4H), 1.19 (s, 3H), 1.07 (s, 3H), 0.96 (s, 2H), 0.86 (s, 2H), 0.63 (s, 1H), 0.47 (s, 1H); 13 C NMR (125 MHz, CDCl 3 ) δ: 186.69, 182.32, 173.65, 167.69, 158.62, 153.40, 150.10, 139.29, 130.08, 129.38, 127.52, 125.46, 120.13, 119.55, 118.83, 115.34, 112.09, 109.33, 44.97, 44.31, 40.78, 40.17, 39.53, 36.46, 35.29, 34.82, 33.31, 32.90, 31.65, 31.07, 30.68, 29.80, 28.71, 20.34, 18.70. HRESIMS (m / z) calcd for C 35 H 41 NO 5 [M + H] + : 556.3065, Found: 556.2694.

[0042] Compound 2 1 H NMR (500 MHz, CDCl 3 ) δ: 10.39 (s, 1H), 7.76 (s, 1H), 7.50 (d, J = 18.0 Hz, 4H), 6.81 (s, 1H), 6.70 (s, 1H), 6.48 (s, 1H), 6.29 (s, 1H), 2.47 (d, J = 13.5 Hz, 1H), 2.24 (d, J = 9.0 Hz, 1H), 2.10 (s, 1H), 1.93 (s, 1H), 1.84 (s, 1H), 1.74 (s, 1H), 1.68 (d, J = 7.0 Hz, 1H), 1.56 (s, 5H), 1.42 (s, 4H), 1.34 (s, 1H), 1.28 (s, 1H), 1.25 (s, 3H), 1.22 (s, 2H), 1.08 (s, 3H), 0.98 (s, 1H), 0.61 (s, 3H); 1313C NMR (125 MHz, CDCl 3 ) δ: 186.52, 182.53, 174.18, 167.23, 157.57, 153.59, 152.93, 150.18, 142.61, 137.96, 130.01, 125.26, 119.77, 112.26, 111.98, 109.45, 105.95, 44.92, 44.15, 40.55, 40.05, 39.33, 36.42, 35.31, 34.49, 33.33, 32.72, 31.94, 31.59, 31.45, 31.17, 30.64, 30.20, 29.71, 29.46, 28.71, 22.71, 20.02, 18.79. HRESIMS (m / z) calcd for C 39 H 43 NO 6 [M + H] + : 622.3170, Found: 622.1243.

[0043] Compound 3 1 1H NMR (500 MHz, CDCl 3 ) δ: 10.91 (s, 1H), 7.71 (s, 4H), 7.49 (s, 1H), 7.32 (d, J = 19.5 Hz, 1H), 6.97 (s, 1H), 6.90 (s, 1H), 6.76 (s, 1H), 6.11 (s, 1H), 1.90 (d, J = 11 Hz, 1H), 1.81 (d, J = 13 Hz, 2H), 1.70 (s, 2H), 1.55 (d, J = 15.5 Hz, 3H), 1.45 (s, 2H), 1.37 (s, 1H), 1.25 (s, 3H), 1.09 (s, 2H), 0.87 (s, 6H), 0.72 (s, 3H), 0.64 (s, 3H), 0.57 (s, 1H); 13 13C NMR (125 MHz, CDCl 3)δ: 188.42, 186.52, 178.26, 173.94, 160.01, 153.45, 150.15, 143.05, 142.24, 134.22, 128.28, 127.92, 127.25, 125.50, 123.95, 123.67, 119.96, 112.80, 44.73, 44.19, 41.02, 39.46, 39.24, 36.09, 35.41, 34.91, 33.26, 32.83, 31.87, 31.58, 31.10, 30.74, 30.65, 30.09, 29.71, 29.48, 28.13, 20.19, 17.96. HRESIMS (m / z) calcd for C 39 H 43 NO 5 S[M + H] + : 638.2942, Found: 638.3628.

[0044] Example 2

[0045] Inhibitory effects of norzelanic aldehyde Schiff base derivatives on the proliferation of HCT116, SKOV3 and HepG2 cells:

[0046] (1) Cell seeding: Digest HCT116, SKOV3 and HepG2 cells in the exponential growth phase, make a suspension with McCoy’s 5A / DMEM medium containing 10% calf serum, and inoculate the tumor cells at a seeding cell number of 5×10 4 cells / mL into a 96-well culture plate, add 100 μL to each well, and culture in an incubator at 37°C and 5% CO 2 for 24 h.

[0047] (2) Drug addition: Prepare a stock solution of the test compound in DMSO at a certain concentration, and then prepare solutions of 30 μM, 20 μM, 10 μM, 5 μM, and 1 μM respectively with McCoy’s 5A / DMEM medium containing 10% calf serum, inoculate them into the 96-well cell culture plate, set three parallel replicates for each concentration, and continue to culture in an incubator at 37°C and 5% CO 2 for 72 h, and observe the cell morphology and growth changes under an inverted microscope.

[0048] (3) Termination of culture: First, prepare a mixture containing 10% CCK-8. Dilute 450 μL of CCK-8 to 4500 μL with serum-free medium, and then add 100 μL of the CCK-8 and medium mixture to each well. Incubate at 37°C and 5% CO 2After culturing in an incubator for 1 h, the absorbance A value at a wavelength of 450 nm was measured on an enzyme-linked immunosorbent assay (ELISA) reader, and the inhibition rate was calculated according to the following formula: Inhibition rate (%) = (Average absorbance A value of the control group - Average absorbance A value of the drug-added group) / (Average absorbance A value of the control group - Average absorbance A value of the blank group) × 100%; the control group was the culture well with a concentration of 0 of norzelanidene Schiff base derivatives and added dimethyl sulfoxide (DMSO), and the blank group was the culture well with medium and CCK-8 solution but without cells. Data statistical analysis was performed using GraphPad Prism 6, and IC 50 .

[0049] The results of the CCK-8 experiment are shown in Table 1, indicating that norzelanidene Schiff base derivatives have an obvious effect of inhibiting the proliferation of HCT116, SKOV3, and HepG2 cells. Among them, norzelanidene Schiff base derivatives have the largest inhibition gradient on the colon cancer cell line HCT116 and have a relatively strong inhibitory effect, which can be further used for the research and development of new anti-colon cancer drugs.

[0050] Table 1

[0051]

[0052] Example 3

[0053] Cell cycle detection by flow cytometry

[0054] (1) Cell seeding: Digest HCT116 cells in the exponential growth phase and make a suspension with McCoy's 5A medium containing 10% fetal bovine serum. Seed the tumor cells at a density of 5×10 5 cells / mL in a 6-well culture plate, add 2 mL to each well, and culture in an incubator at 37 °C and 5% CO 2 for 24 h.

[0055] (2) Drug addition: Prepare a stock solution of the test compound with DMSO at a certain concentration, and then prepare solutions of compound 1 at 3 μM and 6 μM, compound 2 at 2 μM and 8 μM, and compound 3 at 15 μM and 60 μM with McCoy's 5A medium containing 10% fetal bovine serum and inoculate them into a 6-well cell culture plate. Set three parallel replicates for each concentration and continue to culture in an incubator at 37 °C and 5% CO 2 for 24 h. Observe the cell morphology and growth changes under an inverted microscope.

[0056] (3) Cell collection: Wash the cells once with PBS, centrifuge at 1500 rpm for 5 min to collect, and adjust the cell concentration to 1×10 6 / mL, take 1 mL of single-cell suspension. After centrifuging the prepared single-cell suspension, remove the supernatant, wash it once with PBS, centrifuge again, and discard the supernatant.

[0057] (4) Staining: Add 1 mL of DNA Staining solution, resuspend the cells, then add 10 μL of PI staining solution and mix well. Incubate in the dark at room temperature for 30 min.

[0058] (5) Detection by flow cytometry: Select the lowest sample loading speed and record the red fluorescence at an excitation wavelength of 488 nm.

[0059] The flow cytometry results of Compound 1 are as Figure 1 shown. Compared with the control group, Compound 1 at a concentration of 6 μM caused a significant 1.61-fold increase in the G1 phase and a significant 15.62% decrease in the G2 phase. The proportion of cells in the G1 phase increased from 33.2% (control group) to 51.1% (3 μM) and 53.4% (6 μM). At the same time, the proportion of cells in the G2 phase decreased from 23.3% (control group) to 10.2% (3 μM) and 7.68% (6 μM). The percentages of cells in the G1 and G2 phases showed an increasing trend with the increase in the concentration of Compound 1. The results indicate that Compound 1 affects cell cycle arrest in the G1 phase. Compared with the control group, 10 nM of PTX alone could significantly inhibit cells from entering the G2 phase. However, when combined with Compound 1, the proportion of G1-phase cells decreased from 51.5% (3 μM) to 49.4% (6 μM). The proportion of S-phase cells decreased from 29.7% (3 μM) to 25.5% (6 μM), and the proportion of G2-phase cells increased from 20.3% (3 μM) to 23.5% (6 μM). The results indicate that the combination of Compound 1 and PTX affects cell cycle arrest in the G2 phase.

[0060] The flow cytometry results of Compound 2 are as Figure 2As shown, compared with the control group, when the concentration was 8 μM, the G1-phase cells increased significantly by 1.28-fold. When the concentration was 2 μM, the S-phase cells increased significantly by 2.8%. The proportion of G1-phase cells rose from 33.2% (control group) to 35.8% (2 μM) and 42.4% (8 μM); the proportion of G2-phase cells decreased from 23.3% (control group) to 18.5% (2 μM) and 12.5% (8 μM). The percentages of G1-phase and G2-phase cells showed an increasing trend with the increase in the concentration of Compound 2. The results indicate that Compound 2 affects the S-phase arrest of the cell cycle. Compared with the control group, 10 nM of PTX alone could significantly arrest the cells in the G2-phase. However, when combined with Compound 2, the proportion of G1-phase cells rose from 33.2% (control group) to 43.8% (2 μM) and 39.7% (8 μM); the proportion of S-phase cells decreased from 43.3% (control group) to 33.2% (2 μM) and 39.8% (8 μM). The results show that compared with PTX alone, Compound 2 combined with PTX affects the G1-phase cell cycle arrest and changes the arrest phase.

[0061] The flow cytometry results of Compound 3 are as Figure 3 shown. Compared with the control group, in Compound 3 at a concentration of 60 μM, the G1-phase cells increased significantly by 1.41-fold, and the S-phase cells decreased significantly by 8.3%. The proportion of cells in the G1-phase increased from 33.2% (control group) to 39.1% (15 μM) and 46.8% (60 μM). At the same time, the proportion of cells in the G2-phase decreased from 23.3% (control group) to 17.1% (15 μM) and 16.5% (60 μM). The percentages of G1-phase and G2-phase cells showed an increasing trend with the increase in the concentration of Compound 3. The results indicate that Compound 3 affects the G1-phase arrest of the cell cycle. Compared with the control group, 10 nM of PTX alone could significantly arrest the cells in the G2-phase. When combined with Compound 3, the proportion of G1-phase cells rose from 33.2% (control group) to 42.7% (15 μM) and 46.1% (60 μM); the proportion of S-phase cells decreased from 43.3% (control group) to 37% (15 μM) and 32.3% (60 μM). The results show that Compound 3 combined with PTX has an impact on the G1-phase arrest of the cell cycle.

[0062] Example 4

[0063] Flow cytometry detection of cell apoptosis

[0064] (1) Seeding cells: Digest HCT116 cells in the exponential growth phase, make a suspension with McCoy’s 5A medium containing 10% fetal bovine serum, and seed the tumor cells at a density of 1×10 5 cells / mL in a 6-well culture plate, add 2 mL to each well, and culture at 37 °C and 5% CO 2Culture in an incubator for 12 h.

[0065] (2) Drug addition: The test compound was added with DMSO to prepare a stock solution of a certain concentration, and then compound 1 was prepared into 3 μM and 6 μM solutions, compound 2 was prepared into 2 μM and 8 μM solutions, and compound 3 was prepared into 15 μM and 60 μM solutions in Mccoy's 5A medium containing 10% calf serum. The solutions were inoculated into a 6-well cell culture plate, and three parallel wells were set for each concentration. The cells were incubated at 37°C and 5% CO. 2 The cells were cultured in the incubator for 48 h, and the cell morphology and growth changes were observed under an inverted microscope.

[0066] (3) Cell collection: Wash the cells once with PBS, collect them by centrifugation at 1500 rpm for 5 min, and adjust the cell concentration to 1×10 6 / mL, take 1mL of single cell suspension. After centrifugation of the prepared single cell suspension, remove the supernatant.

[0067] (4) Staining: Add 500 μL 1× Binding Buffer to resuspend the cells, add 5 μL Annexin V and 10 μL PI, and incubate at room temperature in the dark for 5 min.

[0068] (5) On-machine detection: record the red fluorescence at the excitation wavelength of 488 nm and 650 nm.

[0069] The flow cytometry results of compound 1 are shown in Figure 4 As shown, compound 1 can significantly induce apoptosis of HCT116 cells in a dose-dependent manner. Taking untreated cells as the control group, the early apoptosis rate induced by compound 1 gradually decreased from 37.80% (control group) to 7.31% (3μM) and 8.05% (6μM). After treatment with 3μM and 6μM concentrations of compound 1, the late apoptosis rate changed from 0 to 15.00% and 31.40%, respectively. Compared with the use of 10nM PTX alone, compound 1 (3μM and 6μM) worked in combination with PTX for 48h. The apoptosis rate of HCT116 cells increased significantly in a dose-dependent manner, and the late apoptosis rates were 26.3% and 57.4%, respectively. The apoptosis ratios of HCT116 cells treated with compound 1 combined with PTX were 40.90% (10nM PTX alone), 33.76% (10nM PTX+3μM compound 1) and 72.60% (10nM PTX+6μM compound 1), respectively.

[0070] The flow cytometry results of compound 2 are shown in Figure 5As shown, compound 2 can significantly induce apoptosis in HCT116 cells in a dose-dependent manner. Using untreated cells as the control group, the early apoptosis rate of compound 2 gradually decreased from 41.30% (control group) to 8.88% (2 μM) and 11.40% (8 μM). After treatment with compound 2 at concentrations of 2 μM and 8 μM, the late apoptosis rates changed from 0 to 20.50% and 59.20% respectively. Compared with the treatment with PTX alone at 10 nM, compound 2 (2 μM and 8 μM) was combined with PTX and acted on HCT116 cells for 48 h. The apoptosis rate of HCT116 cells increased significantly in a dose-dependent manner, and the late apoptosis rates were 35.0% and 61.3% respectively. The apoptosis proportion of HCT116 cells after treatment with 10 nM PTX alone was 40.90%, the combined apoptosis proportion of 10 nM PTX and 2 μM compound 2 was 43.78%, and the combined apoptosis proportion of 10 nM PTX and 8 μM compound 2 was 75.00%.

[0071] The flow cytometry results of compound 3 are as Figure 6 As shown, compound 3 can significantly induce apoptosis in HCT116 cells in a dose-dependent manner. Using untreated cells as the control group, the early apoptosis rate of compound 3 gradually decreased from 60.80% (control group) to 13.2% (15 μM) and 5.44% (8 μM). After treatment at concentrations of 15 μM and 60 μM, the late apoptosis rates changed from 1.06% to 65.60% and 80.10% respectively. Compared with the treatment with 10 nM PTX alone, treating HCT116 cells with compound 3 (15 μM and 60 μM) combined with PTX for 48 h significantly increased the apoptosis rate in a dose-dependent manner. The late apoptosis rates of the combination of 15 μM and 60 μM compound 3 with PTX were 24.4% and 57.8% respectively. The apoptosis proportions of HCT116 cells treated with the combination of compound 3 and PTX were 40.9% (10 nM PTX), 34.7% (10 nM PTX + 15 μM compound 3), and 87.5% (10 nM PTX + 60 μM compound 3) respectively.

[0072] In summary, the above examples disclose 3 norzelanamine Schiff base derivatives. Among them, compound 2 has higher activity than norzelanamine at the cellular level. The synthesis conditions of norzelanamine Schiff base derivatives are mild, insensitive to water, oxygen, etc.; the reaction raw materials are easily available, inexpensive, the operation is simple, the products are easy to separate and purify, the yield is high, and it is easy for industrial production, etc.

[0073] The proliferation inhibitory effects of norzelanic aldehyde Schiff base derivatives on tumor cell lines were determined by the CCK8 method. The results showed that most of the compounds had significant anti-cancer activities against cancer cell lines such as human colon cancer cell line HCT116, human ovarian cancer cell line SKOV3, and human liver cancer cell line HepG2. These compounds had novel structures.

[0074] Flow cytometry detection showed that compound 2 had a cell cycle arrest effect, causing cancer cells HCT116 to stay in the S phase, thus achieving an anti-tumor effect and having an effect of inducing apoptosis of cancer cells.

[0075] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention can have various changes and modifications. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A Schiff base derivative of norzelaminaldehyde having a structural formula as shown in Formula 1: In Formula 1, R is selected from one of the following substituents:

2. The method for preparing the Schiff base derivatives of norzelaminaldehyde according to claim 1, characterized in that: The synthesis route of the Schiff base derivatives of norzelaminaldehyde is shown in Formula 2:

3. Use of the norzelaminaldehyde Schiff base derivative or its pharmaceutically acceptable salt according to claim 1 in the preparation of anticancer drugs.

4. The use according to claim 3, characterized in that: Such cancers include colon cancer, ovarian cancer and liver cancer.

5. The use according to claim 3, characterized in that: The medicine uses the norzelaminaldehyde Schiff base derivative or its pharmaceutically acceptable solvate as an active ingredient, with a weight percentage of 5% to 100%.

6. The use according to claim 3, characterized in that: The medicament includes a pharmaceutically acceptable carrier or diluent.

7. The use according to claim 3, characterized in that: The dosage form of the drug is selected from at least one of tablets, capsules, granules, pellets, suspensions, syrups, enteric-coated preparations, emulsion suspensions and injections.

8. An anti-cancer pharmaceutical composition, characterized in that: It comprises a Schiff base derivative of norzelaminaldehyde or a pharmaceutically acceptable solvate thereof as an active ingredient, wherein the structural formula of the Schiff base derivative of norzelaminaldehyde is as described in claim 1; or further comprises a pharmaceutically acceptable carrier or diluent.

9. The anti-cancer pharmaceutical composition according to claim 8, characterized in that: The weight percentage of the norzelaminaldehyde Schiff base derivative or its pharmaceutically acceptable solvate is 5% to 100%.

10. Use of the norzelaminaldehyde Schiff base derivative or a pharmaceutically acceptable salt thereof according to claim 1 in the preparation of HCT116, SKOV3, HepG2 cell proliferation inhibitors.

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