A Schiff base acylhydrazone zinc complex and its synthesis method and use

Abstract

The present invention belongs to the technical field of anti-tumor drug research, and specifically relates to a Schiff base acylhydrazone zinc complex, its synthesis method and use. The chemical formula of the zinc complex is [ZnL(AC)]2, and the structural formula is shown in Formula I. In the chemical formula, L is 3-(1-(salicylhydrazone)ethyl)pyridine. The test results show that the zinc complex exhibits quite significant anti-cancer activity against the human non-small cell lung cancer cell line A549, proving that the Schiff base acylhydrazone zinc complex is an ideal new anti-cancer drug. The Schiff base acylhydrazone zinc complex prepared by the present invention has a definite structure, low toxicity and good anti-cancer activity. #imgabs0#

Description

Technical Field

[0001] The invention belongs to the technical field of anti-tumor drug research, and specifically relates to a Schiff base acylhydrazone zinc complex and a synthesis method and application thereof. Background Art

[0002] Since the development of cisplatin, the research of metal-based anti-tumor drugs has opened up a new field. However, platinum drugs have shortcomings such as drug resistance and toxic side effects that are difficult to overcome. Breaking out of the limitations of traditional platinum drugs and developing potential new drug molecules, especially new non-platinum metal complexes, has become a hot research topic that humans are eager to study, in order to fill the gap in tumor chemotherapy, including minimizing adverse side effects, overcoming drug resistance problems, and expanding the range of activity to more tumor types. Schiff bases are a class of compounds containing imine or azomethine groups, usually formed by simple condensation of primary amines and carbonyl groups, and are an important class of organic ligands in the synthesis of new drug molecules. Acylhydrazone compounds are a special class of Schiff base compounds with a wide range of uses. The intermolecular hydrogen bonds formed by the O and N atoms in their molecules make them highly stable, and they can also combine with certain substances in the body through intermolecular hydrogen bonds, thereby inhibiting many physiological and chemical processes in the body and showing strong biological activities, such as antibacterial, anti-cancer, anti-viral, anti-inflammatory, anti-tuberculosis, etc., and have a wide range of applications in medicine. It has been reported that some acylhydrazone compounds have been used as antitumor agents.

[0003] Zinc complexes cannot be ignored in the search for anticancer metal drug substitutes for platinum derivatives. As the second largest trace element in the human body, zinc metals are extremely important for the normal function of the human body. Zinc derivatives are ubiquitous in thousands of proteins and enzymes due to their chemical properties, especially the lack of redox activity. They also have the ability to support coordination types of different geometric structures and promote ligand exchange. Similar to other trace elements, when the balance of zinc in the body is disrupted, it can lead to various diseases and may be related to the occurrence of cancer in some cases. For example, zinc deficiency can inhibit immune function because zinc plays a vital role in the immune system through cell proliferation and RNA and DNA synthesis, and is necessary for the development of T lymphocytes. In addition to its physiological effects, zinc also has therapeutic and preventive effects on infectious diseases. Compared with other metal-based drugs, zinc complexes are generally less toxic and have fewer side effects, and have been regarded as potential anti-tumor drugs.

[0004] Therefore, the present invention aims to provide a Schiff base acylhydrazone zinc complex with better tumor treatment effect and high safety. Summary of the invention

[0005] Based on the above technical problems, the present invention provides a Schiff base acylhydrazone zinc complex, whose chemical formula is [ZnL(AC)]2, wherein L is 3-(1-(salicylicylhydrazone)ethyl)pyridine. The zinc complex has a simple synthesis process, low cost, easy control of chemical components, good repeatability and high yield.

[0006] The first object of the present invention is to provide a Schiff base acylhydrazone zinc complex, the chemical formula of which is [ZnL(AC)]2, and the structural formula is shown in Formula I:

[0007]

[0008] In the chemical formula, L is 3-(1-(salicyloylhydrazone)ethyl)pyridine.

[0009] Preferably, the crystal of the zinc complex belongs to the monoclinic system, the space group is P21 / c, and the unit cell parameters are β=108.759(9)°, the unit cell volume is Z=2,Dc=1.618g / cm 3 .

[0010] The second object of the present invention is to provide a method for synthesizing the zinc complex, comprising the following steps:

[0011] The salicylic acid hydrazide and 3-acetyl pyridine are mixed for condensation reaction to obtain the ligand 3-(1-(salicylic acid hydrazone)ethyl)pyridine;

[0012] The 3-(1-(salicyloylhydrazone)ethyl)pyridine is mixed with zinc acetate metal salt for coordination reaction to obtain colorless crystals, which are the zinc complex.

[0013] Preferably, the molar ratio of salicylic acid hydrazide to 3-acetylpyridine is 1:1 to 1.2.

[0014] Preferably, the molar ratio of the 3-(1-(salicyloylhydrazone)ethyl)pyridine to the zinc acetate metal salt is 1:1-2.

[0015] Preferably, the condensation reaction is carried out at 65-75° C. for 3-6 hours.

[0016] Preferably, the coordination reaction is carried out at 70-80° C. for 36-72 hours.

[0017] The third object of the present invention is to provide a use of the zinc complex or zinc complex crystal in the preparation of anti-tumor drugs.

[0018] Preferably, the tumor is liver cancer, breast cancer or lung cancer.

[0019] The fourth object of the present invention is to provide an anti-tumor drug, which includes the zinc complex and pharmaceutically acceptable excipients.

[0020] Compared with the prior art, the beneficial effects of the present invention are:

[0021] 1. The present invention synthesizes Schiff base acylhydrazone zinc complex [ZnL(AC)]2 for the first time by using Schiff base acylhydrazone ligand 3-(1-(salicyloylhydrazone)ethyl)pyridine and zinc acetate by solvothermal method. The test results show that the zinc complex has an inhibitory effect on human liver cancer cell line SMMC-7721, human breast cancer cell line MDA-MB-231 and human non-small cell lung cancer cell line A549, especially on human non-small cell lung cancer cell line A549, showing quite significant anticancer activity, which is more active than the positive control drug cisplatin, and has the ability to inhibit cancer cell invasion and migration, proving that the Schiff base acylhydrazone zinc complex is an ideal new anticancer drug.

[0022] 2. The synthesis process of the Schiff base acylhydrazone zinc complex provided by the present invention is simple, low in cost, easy to control the chemical components, good in repeatability and high in yield. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 It is the crystal structure diagram of the smallest asymmetric unit of the zinc complex;

[0024] Figure 2 This is the infrared image of the zinc complex;

[0025] Figure 3 is the PXRD pattern of the zinc complex, (a) fitting peak, (b) measured peak;

[0026] Figure 4 is the TG-GSC thermogravimetric diagram of the zinc complex;

[0027] Figure 5 is the UV spectrum of the zinc complex in Tris-HCl-NaCl buffer solution;

[0028] Figure 6 This is a statistical analysis chart of the scratch test results of zinc complexes on human lung cancer A549 cells; * indicates p<0.05, ** indicates p<0.01, *** indicates p<0.001, **** indicates p<0.0001;

[0029] Figure 7 This is the flow cytometry result of zinc complex blocking the cycle of human lung cancer A549 cells;

[0030] Figure 8 This is the flow cytometry result of the effect of zinc complexes on apoptosis of human lung cancer A549 cells; *p<0.05, **p<0.01, ***p<0.001. DETAILED DESCRIPTION

[0031] The present invention will be further described below with reference to specific embodiments, but is not limited thereto.

[0032] Example 1

[0033] A Schiff base acylhydrazone zinc complex is prepared according to the following steps:

[0034] Salicylic acid hydrazide and 3-acetyl pyridine were mixed in a molar ratio of 1:1.1, and the solvent was in anhydrous ethanol solution, and the reaction was carried out at 70°C for 4 hours to obtain the ligand 3-(1-(salicylic acid hydrazone)ethyl)pyridine. The specific synthesis route is as follows:

[0035] The ligand 3-(1-(salicyloylhydrazone)ethyl)pyridine and zinc acetate Zn(AC)2·2H2O were mixed in a molar ratio of 1:1 and dissolved in 3 mL of anhydrous ethanol solution. The entire mixed system was placed in a polytetrafluoroethylene stainless steel reactor and reacted at 70°C for 48 hours. The reaction solution was slowly cooled to room temperature to obtain colorless block crystals, which were filtered and dried at room temperature. Yield: 66%. The specific synthesis route of the zinc complex is as follows:

[0036]

[0037] Example 2

[0038] A Schiff base acylhydrazone zinc complex is prepared according to the following steps:

[0039] Salicylic acid hydrazide and 3-acetyl pyridine were mixed in a molar ratio of 1:1.2, the solvent was in anhydrous ethanol solution, and the reaction was carried out at 70° C. for 4 hours to obtain the ligand 3-(1-(salicylic acid hydrazone)ethyl)pyridine;

[0040] The ligand 3-(1-(salicyloylhydrazone)ethyl)pyridine and zinc acetate Zn(AC)2·2H2O were mixed in a molar ratio of 1:1 and dissolved in 3 mL of anhydrous ethanol solution. The entire mixed system was placed in a polytetrafluoroethylene stainless steel reactor and reacted at 70°C for 48 hours. The reaction solution was slowly cooled to room temperature to obtain colorless block crystals, which were filtered and dried at room temperature. Yield: 66%.

[0041] The synthetic route is the same as that of Example 1.

[0042] Example 3

[0043] A Schiff base acylhydrazone zinc complex is prepared according to the following steps:

[0044] Salicylic acid hydrazide and 3-acetyl pyridine were mixed in a molar ratio of 1:1.1, the solvent was in anhydrous ethanol solution, and the reaction was carried out at 70° C. for 4 hours to obtain the ligand 3-(1-(salicylic acid hydrazone)ethyl)pyridine;

[0045] The ligand 3-(1-(salicyloylhydrazone)ethyl)pyridine and zinc acetate Zn(AC)2·2H2O were mixed in a molar ratio of 1:2 and dissolved in 3 mL of anhydrous ethanol solution. The entire mixed system was placed in a polytetrafluoroethylene stainless steel reactor and reacted at 70°C for 48 hours. The reaction solution was slowly cooled to room temperature to obtain colorless block crystals, which were filtered and dried at room temperature. Yield: 64%.

[0046] The synthetic route is the same as that of Example 1.

[0047] Example 4

[0048] A Schiff base acylhydrazone zinc complex is prepared according to the following steps:

[0049] Salicylic acid hydrazide and 3-acetyl pyridine were mixed in a molar ratio of 1:1.2, the solvent was in anhydrous ethanol solution, and the reaction was carried out at 70° C. for 4 hours to obtain the ligand 3-(1-(salicylic acid hydrazone)ethyl)pyridine;

[0050] The ligand 3-(1-(salicyloylhydrazone)ethyl)pyridine and zinc acetate Zn(AC)2·2H2O were mixed in a molar ratio of 1:2 and dissolved in 3 mL of anhydrous ethanol solution. The entire mixed system was placed in a polytetrafluoroethylene stainless steel reactor and reacted at 70°C for 48 hours. The reaction solution was slowly cooled to room temperature to obtain colorless block crystals, which were filtered and dried at room temperature. Yield: 64%.

[0051] The synthetic route is the same as that of Example 1.

[0052] Example 5

[0053] A Schiff base acylhydrazone zinc complex is prepared according to the following steps:

[0054] Salicylic acid hydrazide and 3-acetyl pyridine were mixed in a molar ratio of 1:1.2, the solvent was in anhydrous ethanol solution, and the reaction was carried out at 70° C. for 4 hours to obtain the ligand 3-(1-(salicylic acid hydrazone)ethyl)pyridine;

[0055] The ligand 3-(1-(salicyloylhydrazone)ethyl)pyridine and zinc acetate Zn(AC)2·2H2O were mixed in a molar ratio of 1:2 and dissolved in 3 mL of anhydrous ethanol solution. The entire mixed system was placed in a polytetrafluoroethylene stainless steel reactor and reacted at 70°C for 36 hours. The reaction solution was slowly cooled to room temperature to obtain colorless block crystals, which were filtered and dried at room temperature. Yield: 63%.

[0056] The synthetic route is the same as that of Example 1.

[0057] Example 6

[0058] A Schiff base acylhydrazone zinc complex is prepared according to the following steps:

[0059] Salicylic acid hydrazide and 3-acetyl pyridine were mixed in a molar ratio of 1:1.2, the solvent was in anhydrous ethanol solution, and the reaction was carried out at 70° C. for 4 hours to obtain the ligand 3-(1-(salicylic acid hydrazone)ethyl)pyridine;

[0060] The ligand 3-(1-(salicyloylhydrazone)ethyl)pyridine and zinc acetate Zn(AC)2·2H2O were mixed in a molar ratio of 1:2 and dissolved in 3 mL of anhydrous ethanol solution. The entire mixed system was placed in a polytetrafluoroethylene stainless steel reactor and reacted at 80°C for 72 hours. The reaction solution was slowly cooled to room temperature to obtain colorless block crystals, which were filtered and dried at room temperature. Yield: 62%.

[0061] The synthetic route is the same as that of Example 1.

[0062] Example 7

[0063] A Schiff base acylhydrazone zinc complex is prepared according to the following steps:

[0064] Salicylic acid hydrazide and 3-acetyl pyridine were mixed in a molar ratio of 1:1.2, the solvent was in anhydrous ethanol solution, and the reaction was carried out at 75°C for 3 hours to obtain the ligand 3-(1-(salicylic acid hydrazone)ethyl)pyridine;

[0065] The ligand 3-(1-(salicyloylhydrazone)ethyl)pyridine and zinc acetate Zn(AC)2·2H2O were mixed in a molar ratio of 1:2 and dissolved in 3 mL of anhydrous ethanol solution. The entire mixed system was placed in a polytetrafluoroethylene stainless steel reactor and reacted at 70°C for 72 hours. The reaction solution was slowly cooled to room temperature to obtain colorless block crystals, which were filtered and dried at room temperature. Yield: 65%.

[0066] The synthetic route is the same as that of Example 1.

[0067] Example 8

[0068] A Schiff base acylhydrazone zinc complex is prepared according to the following steps:

[0069] Salicylic acid hydrazide and 3-acetyl pyridine were mixed in a molar ratio of 1:1.2, the solvent was in anhydrous ethanol solution, and the reaction was carried out at 65° C. for 6 hours to obtain the ligand 3-(1-(salicylic acid hydrazone)ethyl)pyridine;

[0070] The ligand 3-(1-(salicyloylhydrazone)ethyl)pyridine and zinc acetate Zn(AC)2·2H2O were mixed in a molar ratio of 1:2 and dissolved in 3 mL of anhydrous ethanol solution. The entire mixed system was placed in a polytetrafluoroethylene stainless steel reactor and reacted at 70°C for 72 hours. The reaction solution was slowly cooled to room temperature to obtain colorless block crystals, which were filtered and dried at room temperature. Yield: 66%.

[0071] The synthetic route is the same as that of Example 1.

[0072] Since the performance and anti-tumor effect of the Schiff base acylhydrazone zinc complexes prepared in Examples 1 to 8 are substantially the same, the performance and effect of the Schiff base acylhydrazone zinc complex provided by the present invention will be described below using Example 1 as an example.

[0073] The zinc complex was obtained as a single crystal and its crystal structure was measured by X-ray single crystal diffractometer. X-ray single crystal diffraction data showed that the zinc complex belongs to the monoclinic system and the P21 / c(14) space group. Figure 1 As shown in the figure, its minimum asymmetric unit consists of a zinc atom, a ligand and a coordinated acetate. The whole molecule is a dimer composed of two zinc atoms, two ligands and two acetate molecules. The zinc atom is a penta-coordinated irregular tetrahedral coordination model, consisting of three oxygen atoms and two nitrogen atoms. The carbonyl oxygen atom and the Schiff base nitrogen atom of the ligand are connected to the zinc atom. The two oxygen atoms of the acetate are coordinated with the zinc atom in a chelated mode. The two pyridine rings of the ligand are not coplanar and present a certain twist angle.

[0074] Single crystals with better crystal forms were selected for single crystal testing on an X-ray single crystal diffraction instrument, and the crystal structure was analyzed using the Olex2 software.

[0075] The results show that the zinc complex belongs to the monoclinic system, the space group is P21 / c, and the unit cell parameters are β=108.759(9)°, the unit cell volume is Z=2,Dc=1.618g / cm 3 .

[0076] The powder diffractometer data of the zinc complex were measured by X-ray powder diffractometer, and the measured peaks were consistent with the peak positions fitted by the crystal structure data, proving the purity of the zinc complex crystalline powder.

[0077] In addition, its infrared spectrum ( Figure 2 )、XRD of zinc complex( Figure 3 ), proving that the zinc complex is phase-pure and stable at room temperature.

[0078] Thermogravimetric diagram of zinc complex ( Figure 4) demonstrated that the zinc complex can be stable up to 300 degrees.

[0079] The UV spectrum of the zinc complex in Tris-HCl-NaCl buffer solution simulating physiological environment ( Figure 5 ) It can be seen that the zinc complex has two maximum absorption peaks at 293nm and 350nm, and does not change over time, which proves that the zinc complex has good stability under simulated physiological environment conditions.

[0080] Experimental Example 1

[0081] Cytotoxicity of zinc complexes on SMMC-7721, MDA-MB-231, A549 and human normal HK-2 cells

[0082] The cytotoxicity of the ligand, control drug cisplatin and zinc complex to three tumor cells (SMMC-7721, MDA-MB-231, A549) and human normal cell HK-2 was determined by CCK-8 method, namely IC 50 The specific operation process is as follows:

[0083] The cells were plated at 5×10 4 / well were inoculated into 96-well cell culture plates. After the cells were attached to the wall and the growth density was appropriate, the old culture medium was removed and new culture medium containing compound solution was added at a volume of 100 μL / well. All compounds used the same concentration range and gradient: 0 μM, 4 μM, 8 μM, 16 μM, 32 μM, 64 μM, 128 μM. After 24 hours, CCK-8 reagent was added to the culture plate at a volume of 10 μL / well, and the absorbance OD value at 450 nm was measured after incubation at 37°C for 1 to 4 hours.

[0084] The IC values ​​of different compounds on different cells were calculated by statistical analysis software. 50 All data are based on the average of three independent experiments. The experimental results are shown in Table 1.

[0085] Table 1 IC values ​​of zinc complexes, ligands and cisplatin on different tumor cell lines and normal HK-2 cells 50 Numeric

[0086]

[0087] The experimental results show that the zinc complex provided by the present invention has the strongest activity against lung cancer A549 cells, and its IC 50 The value was only 10.40±0.6μM, and it was more effective than the positive control drug cisplatin and ligand.

[0088] Experimental Example 2

[0089] Anti-metastatic effect of zinc complexes on human lung cancer A549 cells

[0090] A549 cells were planted on a 6-well plate and observed under a microscope. When the A549 cells grew to 80%, a straight line was quickly drawn in the middle of each well with a 200 μL pipette tip, the culture medium was discarded, and the cells were washed twice with PBS. The cells were treated with zinc complexes at concentrations of 5, 15, and 30 μM, and incubated in an incubator at 37°C and 5% CO2 for 24 hours. The scratch area was observed under a microscope and photographed at 0 hours and 24 hours after administration.

[0091] The results of the scratch test of zinc complexes on human lung cancer A549 cells are as follows Figure 6 As shown. In the cell scratch experiment, it was found that the healing area of ​​A549 cells was significantly reduced under the action of zinc complexes, indicating that the migration ability of cells was also inhibited. The results showed that zinc complexes can effectively block the healing of A549 tumor cells and have a good inhibitory effect on the migration of human lung cancer cells in vitro.

[0092] In order to study the blocking effect of drugs on cell cycle, human lung cancer A549 cells were treated with different concentrations of Schiff base acylhydrazone zinc complexes for 24 hours, and then digested with trypsin containing 0.25% EDTA, and fixed with 70% anhydrous ethanol at 4°C overnight. The fixed cells were collected by centrifugation, and 100 μL RNaseA was added to each concentration and placed in a 37°C water bath for 30 minutes. Finally, 400 μL PI was added to avoid light and stained for 30 minutes, and the experimental results were detected by upflow cytometer.

[0093] The results show that Figure 7 ), the DNA content in the G0 / G1 phase of the Schiff base acylhydrazone zinc complex increased with the increase of its concentration, indicating that the Schiff base acylhydrazone zinc complex inhibited the cell cycle process of human lung cancer cell A549 by blocking the G0 / G1 phase in a concentration-dependent manner.

[0094] The occurrence of tumors is closely related to the disorder of apoptosis. In order to study whether Schiff base acylhydrazone zinc complexes inhibit the growth of A549 tumor cells by inducing apoptosis, A549 cells were treated with zinc complexes at concentrations of 0, 5, and 10 μM for 24 hours, rinsed twice with pre-cooled PBS buffer solution, added with culture medium containing Annexin membrane protein, and then added with 5 μL of Annexin V-FITC, and continued to be cultured for 15 minutes at 4°C in the dark. Then, 5 μL PI was added and cultured for 10 minutes, and the experimental results were finally measured by flow cytometry.

[0095] Compared with the control group, the proportion of cancer cells in apoptosis in the Schiff base acylhydrazone zinc complex increased significantly, and with the increase of concentration, the apoptosis ratio continued to increase. The experimental results are shown in Figure 8The experimental results show that the Schiff base acylhydrazone zinc complex has a significant apoptosis-inducing effect on human lung cancer A549 cells.

[0096] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A Schiff base acylhydrazone zinc complex, characterized in that: Its chemical formula is [ZnL(AC)]2, and its structural formula is shown in Formula I: In the chemical formula, L is 3-(1-(salicyloylhydrazone)ethyl)pyridinium anion.

2. A method for synthesizing the zinc complex according to claim 1, characterized in that: The following steps are involved: The salicylic acid hydrazide and 3-acetyl pyridine are mixed for condensation reaction to obtain the ligand 3-(1-(salicylic acid hydrazone)ethyl)pyridine; The 3-(1-(salicyloylhydrazone)ethyl)pyridine is mixed with zinc acetate metal salt for coordination reaction to obtain colorless crystals, which are the zinc complex.

3. The synthesis method according to claim 2, characterized in that The molar ratio of salicylic acid hydrazide to 3-acetylpyridine is 1:1-1.

2.

4. The synthesis method according to claim 2, characterized in that The molar ratio of the 3-(1-(salicyloylhydrazone)ethyl)pyridine to the zinc acetate metal salt is 1:1-2.

5. The synthesis method according to claim 2, characterized in that The condensation reaction is carried out at 65-75° C. for 3-6 hours.

6. The synthesis method according to claim 2, characterized in that The coordination reaction is carried out at 70-80° C. for 36-72 hours.

7. Use of the zinc complex according to claim 1 in the preparation of anti-tumor drugs, characterized in that: The tumor is liver cancer, breast cancer or lung cancer.

8. An anti-tumor drug, characterized in that: The medicine comprises the zinc complex according to claim 1 and pharmaceutically acceptable excipients.

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