A pyrimidine bishydrazone tetranuclear Fe(II) complex containing quinoline structure, and a preparation method and application thereof

Abstract

The application discloses a quinoline-structure-containing pyrimidine bishydrazone tetranuclear Fe(II) complex, a preparation method and application thereof, and belongs to the technical field of medicines.The complex is a 4,6-di(quinoline-2-formaldehyde) pyrimidine hydrazone Fe(II) complex, and a molecular formula of the complex is [Fe4(C 24 H 18 N8‑H)2(C 24 H 18 N8‑2H)2](ClO4)2·8CH3OH·2H2O.The complex has a strong scavenging capacity on DPPH free radicals, is much stronger than an antioxidant vitamin E of the same concentration, has inhibiting effects on the proliferation of MCF-7, BGC-823 and BEL-7402 tumor cells, and has a stronger inhibiting activity than cisplatin, and has no cytotoxicity on normal cells.It is shown that the quinoline-structure-containing pyrimidine bishydrazone tetranuclear Fe(II) complex has good in-vitro anti-tumor activity, and is expected to be developed into an anti-tumor drug.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and in particular relates to a pyrimidine dihydrazone tetranuclear Fe(II) complex containing a quinoline structure, its preparation method, and its application. Background Technology

[0002] Cancer is a serious disease that threatens human life and health. Currently, the main treatments for malignant tumors include surgery, radiotherapy, chemotherapy, and molecular targeted therapy, with chemotherapy being one of the most effective methods. However, current chemotherapy drugs generally suffer from poor selectivity, strong drug resistance, and significant side effects. Therefore, developing novel anti-tumor drugs with low toxicity and high selectivity remains a major challenge for researchers. Cisplatin, with its unshakeable position in the field of anticancer therapy, has greatly promoted the development of metal complexes. Metal complex drugs can exert a synergistic effect of ligand and metal, significantly enhancing their anti-tumor activity. Pt(II) complexes have been extensively studied as anticancer drugs, and polynuclear platinum complexes have shown significant therapeutic potential, further promoting research on metal complexes in anti-tumor drugs. Currently, research on anti-tumor active complexes mainly focuses on mononuclear and binuclear complexes; research reports on polynuclear complexes are still limited, and studies on the anti-tumor and antioxidant activities of tetranuclear Fe(II) complexes have not yet been reported.

[0003] N-heterocyclic compounds are an important class of compounds with diverse biological activities, such as antioxidation, antitumor activity, and anti-inflammation. Quinoline compounds can exert excellent antitumor activity as tyrosine kinase (EGFR) inhibitors, c-MET inhibitors, and tubulin polymerization inhibitors. Iron is an important trace element in living organisms; intracellular iron deficiency can lead to anemia, diabetes, heart disease, and damage to the kidneys and liver. Therefore, the successful synthesis of novel pyrimidine dihydrazone tetranuclear Fe(II) complexes is an effective way to find safe and efficient antioxidants and antitumor drugs. Summary of the Invention

[0004] To address the aforementioned technical problems, this invention proposes a pyrimidine dihydrazone tetranuclear Fe(II) complex containing a quinoline structure, its preparation method, and its applications. The preparation method of this complex is simple, and it exhibits good antioxidant and antitumor activity.

[0005] To achieve the above objectives, the present invention provides a quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex, which uses a quinoline-containing pyrimidine dihydrazone derivative as a ligand. The molecular formula of the quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex is [Fe4(C 24 H 18 N8-H)2(C 24 H 18 N8-2H)2](ClO4)2·8CH3OH·2H2O;

[0006] The quinoline-containing pyrimidine dihydrazone derivative is 4,6-bis(quinoline-2-carboxaldehyde)pyrimidinehydrazone, with the molecular formula C0. 24 H 18 N8, the structural formula is shown in equation (1):

[0007]

[0008] Furthermore, the preparation method of the pyrimidine dihydrazone derivative containing a quinoline structure includes the following steps:

[0009] 4,6-Dichloropyrimidine was dispersed in hydrazine hydrate and reacted with stirring at room temperature to obtain crude 4,6-dihydrazinopyrimidine intermediate;

[0010] The crude 4,6-dihydrazinopyrimidine intermediate was washed with a mixed solvent of ethanol and water to obtain the 4,6-dihydrazinopyrimidine intermediate.

[0011] The 4,6-dihydrazinopyrimidine intermediate and quinoline-2-carboxaldehyde were dissolved separately in anhydrous ethanol, mixed, stirred at room temperature, and refluxed in an oil bath. During the reaction, a yellow solid gradually precipitated out.

[0012] After the reflux reaction was completed, the product was filtered, washed, and purified by column chromatography to obtain the pyrimidine dihydrazone derivative containing the quinoline structure.

[0013] Furthermore, in the preparation method of the pyrimidine dihydrazone derivative containing a quinoline structure, the ratio of 4,6-dichloropyrimidine to hydrazine hydrate is 1 mmol: 1 mL, preferably 15 mmol of 4,6-dichloropyrimidine and 15 mL of hydrazine hydrate;

[0014] The molar ratio of the 4,6-dihydrazinopyrimidine intermediate to quinoline-2-carboxaldehyde is 1:2.1, preferably 2 mmol of the 4,6-dihydrazinopyrimidine intermediate and 4.2 mmol of the quinoline-2-carboxaldehyde.

[0015] Furthermore, the preparation method of the quinoline-containing pyrimidine dihydrazone derivative includes the following steps:

[0016] (1) 15 mmol of 4,6-dichloropyrimidine was dispersed in 15 mL of hydrazine hydrate and stirred at room temperature for 1 h to obtain crude 4,6-dihydrazylpyrimidine intermediate;

[0017] (2) The crude 4,6-dihydrazinopyrimidine intermediate was washed with a mixed solvent of ethanol and water to obtain the 4,6-dihydrazinopyrimidine intermediate;

[0018] (3) Dissolve 2 mmol of 4,6-dihydrazinopyrimidine intermediate and 4.2 mmol of quinoline-2-carboxaldehyde in anhydrous ethanol respectively, mix them and stir at room temperature for 2 h, and then reflux in an oil bath. During the reaction, a yellow solid gradually precipitates out.

[0019] (4) After the reflux reaction is completed, the product is filtered, washed with ethanol, and purified by column chromatography to obtain the pyrimidine dihydrazone derivative containing the quinoline structure.

[0020] Furthermore, the quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex belongs to the monoclinic crystal system, space group P21 / c, and its unit cell parameters are as follows: α=90°, β=99.430(2)°, γ=90°,

[0021] A method for preparing the quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex includes the following steps:

[0022] A pyrimidine dihydrazone derivative containing a quinoline ligand was dissolved in a mixed solvent of methanol and dichloromethane to obtain a ligand solution.

[0023] Ferrous perchlorate hexahydrate was added to the ligand solution and stirred until homogeneous;

[0024] The solution was filtered, and the filtrate was allowed to evaporate naturally at room temperature until reddish-black blocky crystals precipitated, which were the pyrimidine dihydrazone tetranuclear Fe(II) complex containing the quinoline structure.

[0025] Furthermore, in the preparation method of the pyrimidine dihydrazone tetranuclear Fe(II) complex containing a quinoline structure, the volume ratio of methanol to dichloromethane in the mixed solvent is 1:1;

[0026] The molar ratio of the pyrimidine dihydrazone derivative containing a quinoline structure to the ferrous perchlorate hexahydrate is 1:1.

[0027] The application of the quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex in the preparation of drugs for scavenging DPPH free radicals.

[0028] The application of the quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex in the preparation of antitumor active drugs.

[0029] Furthermore, the tumor cells are human breast cancer cells MCF-7, human gastric cancer cells BGC-823, and liver cancer cells BEL-7402.

[0030] The application of the quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex in the preparation of antioxidant drugs.

[0031] Compared with the prior art, the present invention has the following advantages and technical effects:

[0032] 1. The ligand 4,6-bis(quinoline-2-carboxaldehyde)pyrimidine hydrazone of this invention has a significantly different structure from the heterocyclic pyrimidine derivatives reported so far. This invention is based on pyrimidine dihydrazone derivatives containing quinoline structures. There are no literature reports on the antioxidant activity of such derivatives and their functional complexes. The ligand synthesis method is simple, convenient to operate, has a high yield, and is easy to industrialize.

[0033] 2. The ligand 4,6-bis(quinoline-2-carboxaldehyde)pyrimidinehydrazone of the present invention is a chelating ligand. Each Fe(II) adopts a six-coordinate manner, chelating and coordinating with the N atom on the pyrimidine ring of the two pyrimidinehydrazone ligands, the N atom on the C=N double bond and the N atom on the quinoline ring, respectively. The four ligands and the four Fe(II) centers are coordinated to form a tetranuclear node-like structure.

[0034] 3. This invention uses 4,6-bis(quinoline-2-carboxaldehyde)pyrimidine hydrazone as a ligand and Fe(II) as the central metal, with a 1:1 feed ratio, to prepare a novel tetranuclear Fe(II) complex of pyrimidine dihydrazone, which effectively solves the problem of binding of quinoline-containing pyrimidine dihydrazone as a ligand to the transition metal Fe(II).

[0035] 4. The Fe(II) complex prepared in this invention, with 4,6-bis(quinoline-2-carboxaldehyde)pyrimidine hydrazone as a ligand, exhibits strong scavenging activity against DPPH free radicals. This indicates that the complex possesses excellent in vitro antioxidant activity.

[0036] 5. Compared with existing technologies, this invention provides a novel method for synthesizing Fe(II) complexes with 4,6-bis(quinoline-2-carboxaldehyde)pyrimidine hydrazone as a ligand. The tetranuclear Fe(II) complex containing a quinoline structure exhibits strong DPPH free radical scavenging ability, far exceeding that of the antioxidant vitamin E at the same concentration. At a concentration of 1 mg / mL, the DPPH free radical scavenging rate reaches 100%, indicating excellent in vitro antioxidant activity and its potential for effective use in the preparation of anti-aging drugs. Simultaneously, the inhibitory effects of the 4,6-bis(quinoline-2-carboxaldehyde)pyrimidine hydrazone ligand and its corresponding Fe(II) complex on the proliferation of three tumor cell lines—human breast cancer MCF-7, human gastric cancer BGC-823, and liver cancer BEL-7402—were studied using the MTT assay, and its cytotoxicity against normal liver cells HL-7702 was evaluated. Experimental results show that the complex inhibits the proliferation of all selected tumor cell lines, with an IC50 concentration of [missing value]. 50The concentrations were 8.61 μM, 5.01 μM, and 7.00 μM, respectively, indicating stronger inhibitory activity than the positive control drug cisplatin, and no cytotoxicity to normal HL-7702 cells. Furthermore, the pyrimidine dihydrazone tetranuclear Fe(II) complex was found to have significantly stronger inhibitory activity against tumor cell proliferation than the ligand 4,6-bis(quinoline-2-carboxaldehyde)pyrimidinehydrazone itself. This indicates that the pyrimidine dihydrazone tetranuclear Fe(II) complex of the present invention possesses good in vitro antitumor activity and holds promise for development into an antitumor drug. Attached Figure Description

[0037] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an undue limitation of the invention. In the drawings:

[0038] Figure 1 The image shows the infrared results of ligand 4,6-bis(quinoline-2-carboxaldehyde)pyrimidinehydrazone in Example 1 of this invention.

[0039] Figure 2 The complex [Fe4(C] in Example 2 of this invention 24 H 18 N8-H)2(C 24 H 18 Infrared results of N8-2H)2](ClO4)2·8CH3OH·2H2O.

[0040] Figure 3 The complex [Fe4(C] in Example 2 of this invention 24 H 18 N8-H)2(C 24 H 18 The main molecular structure diagram of N8-2H)2](ClO4)2·8CH3OH·2H2O.

[0041] Figure 4 The comparative example of the present invention is the complex [Fe4]. II (C 30 H 22 The main molecular structure of N8-H)4](ClO4)4·9CH3CN·CH2Cl2.

[0042] Figure 5 The control example complex [Fe4] II (C 30 H 22 Infrared results of N8-H)4](ClO4)4·9CH3CN·CH2Cl2.

[0043] Figure 6This is a graph showing the scavenging rate of DPPH free radicals by the complex in Example 2 of the present invention, the control complex, and the control BHT.

[0044] Figure 7 The IC50 values ​​of the ligand of Example 1 (the ligand of this invention), the complex of Example 2 (the complex of this invention), the control complex, and the positive control drug cisplatin after 48 hours of treatment in three types of tumor cells were analyzed. 50 Bar chart. Detailed Implementation

[0045] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0046] Unless otherwise specified, the room temperature in the embodiments of the present invention is 25±2℃.

[0047] Example 1

[0048] A method for preparing 4,6-bis(quinoline-2-carboxaldehyde)pyrimidine hydrazone:

[0049] (1) 15 mmol of 4,6-dichloropyrimidine was dispersed in 15 mL (excess) of 80% hydrazine hydrate and stirred at room temperature for 1 h to obtain crude 4,6-dihydrazinopyrimidine intermediate;

[0050] (2) The crude 4,6-dihydrazinopyrimidine intermediate was washed with a mixed solvent of ethanol and water to obtain the 4,6-dihydrazinopyrimidine intermediate;

[0051] (3) Dissolve 2 mmol of 4,6-dihydrazinopyrimidine intermediate and 4.2 mmol of quinoline-2-carboxaldehyde in anhydrous ethanol respectively, mix them, stir at room temperature for 2 h, and reflux in an oil bath at 78 °C. During the reaction, a yellow solid gradually precipitates out.

[0052] (4) After the reflux reaction was completed, the mixture was filtered, washed with ethanol, and purified by column chromatography (petroleum ether:ethyl acetate = 1:1, v:v) to obtain 4,6-bis(quinoline-2-carboxaldehyde)pyrimidine hydrazone with a yield of 54% and a purity of 98%. Its molecular formula is C2. 24 H 18 N8 has the following structural formula:

[0053]

[0054] The infrared spectrum (cm²) of the prepared 4,6-bis(quinoline-2-carboxaldehyde)pyrimidine hydrazone -1 (KBr) such as Figure 1As shown: 3170,3000,1608,1583,1558,1502,1454,1415,1375,1324,1247,1197,1143,1112,985,827,746.

[0055] Example 2

[0056] Using the 4,6-bis(quinoline-2-carboxaldehyde)pyrimidine hydrazone prepared in Example 1 as a ligand, a tetranuclear Fe(II) complex containing a quinoline structure was prepared. The preparation method is as follows:

[0057] (1) Dissolve 0.025 mmol of ligand 4,6-bis(quinoline-2-carboxaldehyde)pyrimidinehydrazone in a mixed solvent of methanol and dichloromethane in a volume ratio of 1:1 to obtain a ligand solution;

[0058] (2) Add 0.025 mmol of ferrous perchlorate hexahydrate to the above ligand solution and stir at room temperature for 2 h.

[0059] (3) Filtration: The filtrate is allowed to evaporate naturally at room temperature. After about two weeks, reddish-black blocky crystals precipitate, which are pyrimidine dihydrazone tetranuclear Fe(II) complexes containing a quinoline structure, with the molecular formula [Fe4(C 24 H 18 N8-H)2(C 24 H 18 The infrared spectrum of the complex is as follows: [N8-2H)2](ClO4)2·8CH3OH·2H2O, Figure 2 As shown (cm) -1 , KBr): 3054, 1631, 1614, 1600, 1517, 1467, 1434, 1423, 1375, 1332, 1278, 1189, 1093, 935, 865, 840, 746.

[0060] The experimental conditions and results of X-ray single-crystal diffraction of the quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex prepared in this embodiment are as follows:

[0061] Single-crystal structure data were determined using a SuperNova Dual diffractometer equipped with a graphite monochromator. A suitable size (0.23 × 0.21 × 0.2 mm) was selected. 3 Crystals were fixed on glass wool to collect crystal data. CuKα rays were used. Using the ω-scanning method, 77,252 diffraction points were collected in the range of 4.014° < θ < 50°, of which 18,935 were independent diffraction points (R). int=0.0748). Empirical absorption correction was performed using the SADABS procedure. The molecular structure of the compound was solved by a direct method and refined using the SHELXTL full-matrix least squares method. All non-hydrogen atoms were refined using anisotropic thermal parameters, and the positions of hydrogen atoms were finally determined using the theoretical hydrogenation method.

[0062] The crystal parameters of the prepared pyrimidine dihydrazone tetranuclear Fe(II) complex single crystals are shown in Table 1, the important bond lengths and bond angles are shown in Table 2, and the main structure of the complex crystals is shown in Table 2. Figure 3 The complex is a monoclinic crystal system, space group P21 / c, with unit cell parameters of... α=90°, β=99.430(2)°, γ=90°,

[0063] Table 1. List of crystal parameters of the complexes prepared in Example 2

[0064]

[0065] Table 2. Main bond lengths and bond angles of the complexes prepared in Example 2.

[0066]

[0067]

[0068] Comparison Example

[0069] A phenyl-modified pyrimidine-quinoline dihydrazone tetranuclear Fe(II) complex, which is a 2-phenyl-4,6-bis(quinoline-2-carboxaldehyde)pyrimidine hydrazone tetranuclear Fe(II) complex with the molecular formula [Fe4] II (C 30 H 22 [N8-H)4](ClO4)4·9CH3CN·CH2Cl2, the main molecular structure is as follows: Figure 4 As shown, the control example complex [Fe4] II (C 30 H 22 The infrared results of N8-H)4](ClO4)4·9CH3CN·CH2Cl2 are shown in the figure. Figure 5 .

[0070] Among them, C 30 H 22 N8 is the ligand 2-phenyl-4,6-bis(quinoline-2-carboxaldehyde), and its structural formula is as follows:

[0071]

[0072] The preparation method of the ligand 2-phenyl-4,6-bis(quinoline-2-carboxaldehyde)pyrimidinehydrazone is as follows:

[0073] (1) 15 mmol of 2-phenyl-4,6-dichloropyrimidine was dissolved in 15 mL (excess) of 80% hydrazine hydrate and stirred at room temperature for 1 h to obtain crude 4,6-dihydrazinopyrimidine intermediate;

[0074] (2) The crude 4,6-dihydrazinopyrimidine intermediate was washed with a mixed solvent of ethanol and water and dried to obtain a white solid powder, 2-phenyl-4,6-dihydrazinopyrimidine intermediate.

[0075] (3) 2 mmol of 2-phenyl-4,6-dihydrazinopyrimidine intermediate and 4.2 mmol of quinoline-2-carboxaldehyde were dissolved in anhydrous ethanol and stirred at room temperature for 2 h. The reaction was carried out under oil bath reflux. During the reaction, a white solid gradually precipitated out.

[0076] (4) After the reflux reaction was completed, the product 2-phenyl-4,6-bis(quinoline-2-carboxaldehyde)pyrimidinehydrazone was obtained by filtration, washing with ethanol, and column chromatography purification (petroleum ether: ethyl acetate = 1:1, v:v).

[0077] The infrared spectrum (cm²) of the prepared 2-phenyl-4,6-bis(quinoline-2-carboxaldehyde)pyrimidinehydrazone -1 , KBr): 3203,3039,1604,1592,1563,4533,1504,1461,1411,1388,1182,1147,1112,825,748,696.

[0078] Using the 2-phenyl-4,6-bis(quinoline-2-carboxaldehyde)pyrimidine hydrazone prepared above as a ligand, a pyrimidine-quinoline dihydrazone Fe(II) complex was prepared:

[0079] (1) Accurately weigh 0.0125 mmol of ligand 2-phenyl-4,6-bis(quinoline-2-carboxaldehyde)pyrimidinehydrazone, dissolve it in a mixed solvent of acetonitrile and dichloromethane with a volume ratio of 1:1 to obtain a ligand solution;

[0080] (2) Accurately weigh 0.025 mmol of ferrous perchlorate hexahydrate, add it to the ligand solution, and stir at room temperature for 2 h;

[0081] (3) Filtration, the filtrate was allowed to stand at room temperature to evaporate, and after one week, reddish-black blocky crystals were precipitated, which is the pyrimidine-quinoline dihydrazone Fe(II) complex, with a yield of 51.2% and a purity of 98%.

[0082] The infrared spectrum of the control example pyrimidine-quinoline dihydrazone Fe(II) complex is as follows: Figure 4 As shown (cm) -1, KBr): 3029, 1666, 1633, 1583, 1563, 1517, 1479, 1459, 1419, 1405, 1340, 1295, 1261 , 1241, 1195, 1168, 1143, 1103, 1002, 985, 931, 908, 865, 827, 782, 767, 750, 696, 624.

[0083] Experimental Example

[0084] The method for determining the in vitro antioxidant activity of this invention is as follows:

[0085] 1. Preparation of reagents

[0086] Preparation of DPPH solution: Dissolve 2 mg of 1,1-diphenyl-2-picrylhydrazine radical (DPPH) in methanol and dilute to 50 mL in a brown volumetric flask. Prepare fresh before use.

[0087] Preparation of Vitamin E solution: Take one Vitamin E capsule (100mg / capsule), cut open the capsule shell, dissolve it in methanol, and dilute to 100mL in a volumetric flask for later use.

[0088] Preparation of sample solutions: Weigh 10 mg each of the sample (the complex from Example 2 of this invention and the control example complex), dissolve in methanol by sonication, and dilute to a 10 mL volumetric flask to obtain a stock solution with a concentration of 1 mg / mL. Then, dilute with methanol sequentially to obtain sample solutions with concentrations of 0.8 mg / mL, 0.6 mg / mL, 0.4 mg / mL, 0.2 mg / mL, 0.1 mg / mL, 0.05 mg / mL, and 0.025 mg / mL. Calculate the sample inhibition rate according to the antioxidant inhibition rate formula (as follows):

[0089] DPPH inhibition rate (%) = [1cm - (A)] i -A j ) / A c ]×100%

[0090] Among them, A i The absorbance values ​​were measured by adding 1 mL of the sample solution of each concentration to 2 mL of DPPH solution; A j The absorbance values ​​measured by adding 1 mL of each concentration of the sample solution to 2 mL of methanol; A c The absorbance value was measured when 1 mL of methanol was added to 2 mL of DPPH solution.

[0091] The measurement of vitamin E in the control complex and positive control was performed according to the measurement method for the sample.

[0092] The antioxidant activities of the complex in Example 2 of this invention, vitamin E, and the control complex are shown in Table 3. The relationship between the DPPH free radical scavenging rate and concentration is shown in the curves. Figure 6 .

[0093] Table 3. Scavenging rates (%) of DPPH free radicals by compounds of different concentrations.

[0094]

[0095]

[0096] The results of in vitro antioxidant experiments showed that the quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex prepared in this invention has a strong scavenging ability against DPPH free radicals, with a scavenging effect far stronger than that of vitamin E at the same concentration, and also significantly stronger than that of the control complex. This indicates that the quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex prepared in this invention has excellent antioxidant properties and is expected to be further used in the preparation of anti-aging drugs.

[0097] The above-mentioned complexes and ligands were used in in vitro antitumor activity assays, and the determination methods are as follows:

[0098] The experiment used the MTT assay, and the absorbance was measured at 570 nm using an enzyme-linked immunosorbent assay (ELISA) reader. Cell viability and IC50 were then calculated. 50 value.

[0099] (1) Cell seeding: Human breast cancer cells MCF-7, human gastric cancer cells BGC-823, human liver cancer cells BEL-7402, and normal cells HL-7702 were used as experimental cells. The number of cells per well was adjusted to 5000 according to Formula 1, and the cells were seeded into 96-well plates. Each plate had a blank control group (only complete culture medium was added, no cells), a negative control group (cells without drug), and drug-added groups (cells containing the ligand and complex of this invention) with drug concentrations of 20 μg / mL, 10 μg / mL, 5 μg / mL, 2.5 μg / mL, and 1.25 μg / mL, respectively, with four replicates per group. The inhibition rate at each concentration was calculated according to Formula 2. The experimental methods for the control complex and the positive control Cisplatin were the same as above.

[0100] (2) After culturing cells for 24 hours and achieving successful adherence and growth, remove them from the incubator and aspirate the original culture medium. Add 200 μL of complete culture medium to the blank group and the control group. For the drug-treated group, add 200 μL of the drug to each well in the order of diluted drug concentration. Add 200 μL of PBS around the perimeter to ensure a humid environment for cell culture.

[0101] (3) Drug action: Place in an incubator and continue culturing for 48 hours. After 48 hours, remove the 96-well plate and add 20 μL of 5 mg / mL MTT to each well (avoid light during the operation).

[0102] (4) Continue culturing in the incubator for 4 hours. Carefully aspirate the original culture medium, avoiding carrying cells out. Add 150 μL of DMSO to each well sequentially, gently blowing the cells to ensure the crystal particles are fully dissolved (avoid generating air bubbles during this process). Shake on a shaker for approximately 10 minutes. Precisely measure the optical density (OD) value of each well at a wavelength of 490 nm using a microplate reader. Finally, calculate the IC50 of the drug. 50 value.

[0103]

[0104] Inhibition rate = (Negative control group - (Drug-treated group - Blank)) / (Negative control group - Blank) Formula 2

[0105] IC50 of the in vitro antitumor activity of the complex of Example 2, the ligand of Example 1, the control complex, and cisplatin of the present invention. 50 (μM) values ​​are shown in Table 4, and the bar chart is shown in […]. Figure 7 .

[0106] Table 4. IC50 of the complexes of Example 2, ligands of Example 1, control complexes, and cisplatin against three types of tumor cells and normal cells in this invention. 50 Value (μM)

[0107]

[0108] Note: "--" indicates that the sample has no inhibitory effect on the experimental cells.

[0109] The results of in vitro antitumor activity assays showed that the quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex prepared in this invention inhibited the proliferation of human breast cancer cells MCF-7, human gastric cancer cells BGC-823, and liver cancer cells BEL-7402. The inhibitory activity was stronger than that of the positive control cisplatin (IC50). 50 The values ​​were 8.61 μM, 5.01 μM, and 7.00 μM, respectively, and it showed no cytotoxicity to normal HL-7702 cells. The control Fe(II) complex only showed inhibitory activity against BEL-7402 liver cancer cells, with an IC50 value of 8.61 μM, 5.01 μM, and 7.00 μM. 50The value was 10.87 μM, showing no inhibitory effect on the other two types of tumor cells, and the overall effect was significantly inferior to the Fe(II) complex of this invention. It was also found that the quinoline-containing pyrimidine dihydrazone tetraphosphate Fe(II) complex of this invention had a stronger inhibitory effect on tumor cells than the ligand 4,6-bis(quinoline-2-carboxaldehyde)pyrimidinehydrazone alone. The IC50 of the ligand in Example 1 of this invention (the ligand of this invention) was... 50 The value is approximately 2 to 5 times that of the tetranuclear Fe(II) complex of Example 2 of this invention (the complex of this invention). This indicates that, in terms of in vitro antitumor activity alone, the quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex synthesized in this invention has a significant advantage. In summary, the tetranuclear nodular Fe(II) complex of this invention exhibits good in vitro antitumor activity, demonstrating a positive synergistic effect after the ligand coordinates with Fe(II) ions. This complex has potential application value and is expected to be used in the preparation of antitumor drugs.

[0110] The above are merely preferred embodiments of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A pyrimidine dihydrazone tetranuclear Fe(II) complex containing a quinoline structure, characterized in that, It uses a pyrimidine dihydrazone derivative containing a quinoline structure as a ligand, and the molecular formula of the pyrimidine dihydrazone tetranuclear Fe(II) complex containing a quinoline structure is [Fe4(C 24 H 18 N8-H)2(C 24 H 18 N8-2H)2](ClO4)2·8CH3OH·2H2O; The quinoline-containing pyrimidine dihydrazone derivative is 4,6-bis(quinoline-2-carboxaldehyde)pyrimidinehydrazone, with the molecular formula C0. 24 H 18 N8, the structural formula is shown in equation (1): Equation (1).

2. The pyrimidine dihydrazone tetranuclear Fe(II) complex containing a quinoline structure according to claim 1, characterized in that, It belongs to the monoclinic crystal system. P Space group 21 / c, cell parameters are a = 15.4542(9) Å, b = 27.1854(16) Å, c = 26.0016(13) Å, α = 90°, β = 99.430(2)°, γ = 90°, V = 10776.4(10) Å 3 .

3. A method for preparing a quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex according to any one of claims 1-2, characterized in that, Includes the following steps: A pyrimidine dihydrazone derivative containing a quinoline ligand was dissolved in a mixed solvent of methanol and dichloromethane to obtain a ligand solution. Ferrous perchlorate hexahydrate was added to the ligand solution and stirred until homogeneous; The solution was filtered, and the filtrate was allowed to evaporate naturally at room temperature until reddish-black blocky crystals precipitated, which were the pyrimidine dihydrazone tetranuclear Fe(II) complex containing the quinoline structure.

4. The method for preparing the quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex according to claim 3, characterized in that, The volume ratio of methanol to dichloromethane in the mixed solvent is 1:1; The molar ratio of the pyrimidine dihydrazone derivative containing a quinoline structure to the ferrous perchlorate hexahydrate is 1:

1.

5. The use of the quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex according to any one of claims 1-2 in the preparation of drugs for scavenging DPPH free radicals.

6. The use of the quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex according to any one of claims 1-2 in the preparation of antitumor active drugs.

7. The use of the quinoline-containing pyrimidine dihydrazone tetranuclear Fe(II) complex according to any one of claims 1-2 in the preparation of antioxidant drugs.

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