Preparation and application of 8-hydroxyquinoline-pyridine anticancer zinc (II) complex

Abstract

The application discloses a kind of 8-hydroxyquinoline-pyridine anticancer zinc (II) complex preparation and application, belong to medical technology field, solve the technical problem of drug resistance of prior art in cisplatin, the method is as follows: Zn (NO3) 2·6H2O, methanol and dichloromethane mixed solution and second ligand are mixed in pressure tube, and cover tightly cover, under 60-70 ℃ 3 days, after reaction, cooling to room temperature, first ligand and triethylamine are added to pressure tube, place in closed environment, under 75-85 ℃ 3 days, cooling to room temperature and stationary, obtain target complex. 50 It is found that the complex has good inhibitory effect on SK-OV-3 / DDP, the IC 50 value is 1.45-10.14 μM, and the toxicity to normal cells HL-7702 is very small, which shows that the complex has selectivity to tumor cells SK-OV-3 / DDP, overcomes the drug resistance of clinical drug, has potential medicinal value, and is expected to be used for the preparation of various antitumor drugs.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and more specifically, to the preparation and application of an 8-hydroxyquinoline-pyridine anticancer zinc(II) complex. Background Technology

[0002] Cancer is one of the diseases that threaten human health. Currently, it can only be treated with radiotherapy, chemotherapy and other methods. However, due to the significant side effects of cisplatin (CDDP) drugs in chemotherapy, the development of non-platinum drugs has become one of the hot topics of research.

[0003] In addition, zinc is one of the trace elements required by the human body and has important physiological functions. It plays a crucial role in growth and development, immunity, and the function of vitamins. Zinc is essential for the synthesis of various enzymes, nucleic acids, proteins, and carbohydrates in the human body. In recent years, zinc complexes have also achieved certain results in anti-tumor research, especially the 20 8-hydroxyquinoline-pyridine DQ1–DQ complexes reported by Qin Qipin's research group. 20 It has a good inhibitory effect on SK-OV-3 / DDP, IC 50 The concentration was below 2.25 ± 0.13 μM, and the toxicity to normal cells was also low, indicating that pyridine compounds reduced the toxicity of 8-hydroxyquinoline zinc complexes to normal cells. On the other hand, studies on the synthesis of zinc complexes and their anticancer activity using 5,7-dichloro-8-hydroxy-2-methylquinoline (H-ClQL) as the first ligand and pyridine derivatives (CY1–CY6) as auxiliary ligands have not been reported in the literature. Summary of the Invention

[0004] One of the objectives of this invention is to provide an 8-hydroxyquinoline-pyridine anticancer zinc(II) complex.

[0005] The second objective of this invention is to provide a method for preparing an 8-hydroxyquinoline-pyridine anticancer zinc(II) complex.

[0006] The third objective of this invention is to provide an application of an 8-hydroxyquinoline-pyridine anticancer zinc(II) complex in the preparation of antitumor drugs.

[0007] The first objective of this invention is achieved through the following technical solution: an 8-hydroxyquinoline-pyridine anticancer zinc(II) complex, the chemical structural formula of which is shown below:

[0008]

[0009] The second objective of this invention is achieved through the following technical solution: a method for preparing an 8-hydroxyquinoline-pyridine anticancer zinc(II) complex, comprising the following steps:

[0010] (1) A mixed solution is obtained by mixing Zn(NO3)2·6H2O, methanol and dichloromethane and a second ligand; the second ligand is 1,10-phenanthroline, 4,7-diphenyl-1,10-phenanthroline, 4,4'-dimethyl-2,2'-bipyridine, 55'-dimethyl-2,2-bipyridine, 4,7-dichloro-1,10-phenanthroline or 2,2-bipyridine;

[0011] (2) The resulting mixed solution was reacted at 60-70℃ for 3 days under sealed conditions;

[0012] (3) After the reaction is completed and cooled to room temperature, the first ligand and triethylamine are added to the pressure-resistant tube; the first ligand is 5,7-dichloro-8-hydroxy-2-methylquinoline;

[0013] (4) Place the pressure-resistant tube in a sealed environment and react at 75-85℃ for 3 days. Cool to room temperature and let stand.

[0014] As a further improvement, in step (1), the mixed solution of methanol and dichloromethane is in a methanol volume: dichloromethane volume = 3:1.

[0015] Further, in step (1), the amount of Zn(NO3)2·6H2O is 1.0 mmol, the amount of the mixed solution of methanol and dichloromethane is 2.0 mmol, and the amount of the second ligand is 1.0 mmol.

[0016] Furthermore, the reaction temperature in step (2) is 65°C.

[0017] Furthermore, in step (3), the amount of the first ligand is 2.0 mmol and the amount of triethylamine is 0.1 mL.

[0018] Furthermore, the reaction temperature in step (4) is 80°C.

[0019] The third objective of this invention is achieved through the application of 8-hydroxyquinoline-pyridine anticancer zinc(II) complexes in the preparation of antitumor drugs.

[0020] The 8-hydroxyquinoline-pyridine anticancer zinc(II) complexes (CYQ1-CYQ6) of this invention have the chemical formulas [Zn(ClQL)2(CY1)](CYQ1), [Zn(ClQL)2(CY2)](CYQ2), [Zn(ClQL)2(CY3)](CYQ3), [Zn(ClQL)2(CY4)](CYQ4), [Zn(ClQL)2(CY5)](CYQ5), and [Zn(ClQL)2(CY6)](CYQ6), and their chemical structural formulas are as follows:Figure 1 As shown.

[0021] Beneficial effects

[0022] Compared with existing technologies, the advantages of this invention are as follows: This invention provides six novel 8-hydroxyquinoline-pyridine anticancer zinc(II) complexes [Zn(ClQL)2(CY1)](CYQ1), [Zn(ClQL)2(CY2)](CYQ2), [Zn(ClQL)2(CY3)](CYQ3), [Zn(ClQL)2(CY4)](CYQ4), [Zn(ClQL)2(CY5)](CYQ5), and [Zn(ClQL)2(CY6)](CYQ6), along with their preparation methods and applications; and investigated their activity and toxicity against the drug-resistant human ovarian cancer cell line SK-OV-3 / DDP and normal HL-7702 cells. Experimental results show that complexes CYQ1-CYQ6 have good inhibitory effects on the drug-resistant ovarian cancer cell line SK-OV-3 / DDP, with an IC50 concentration of […]. 50 The values ​​ranged from 1.45 to 10.14 μM, with the most significant inhibition effect observed in CYQ2, and its IC50 value was [missing value]. 50 The value was 1.45±0.09 μM, which is much higher than that of pyridine derivatives CY1-CY6 and the clinical drug cisplatin (IC50). 50 The concentration was >50 μM, and the toxicity to normal HL-7702 cells was very low, indicating that the complex CYQ2 has tumor selectivity for SK-OV-3 / DDP tumor cells and overcomes drug resistance in clinical use. The novel 8-hydroxyquinoline-pyridine anticancer zinc(II) complexes CYQ1–CYQ6 exhibit superior antitumor activity and have potential pharmaceutical value, showing promise for the preparation of various antitumor drugs. Attached Figure Description

[0023] Figure 1 The CYQ1–CYQ6 structural formula of the target product of this invention;

[0024] Figure 2 This is a synthetic route diagram of the present invention;

[0025] Figure 3 This is a single-crystal structure diagram of CYQ1 of the present invention;

[0026] Figure 4 This is a single-crystal structure diagram of CYQ2 of the present invention;

[0027] Figure 5 This is a single-crystal structure diagram of CYQ3 of the present invention;

[0028] Figure 6 This is a single-crystal structure diagram of CYQ4 of the present invention;

[0029] Figure 7 This is a single-crystal structure diagram of CYQ5 of the present invention;

[0030] Figure 8 This is a single-crystal structure diagram of CYQ6 of the present invention. Detailed Implementation

[0031] The present invention will be further described below with reference to specific embodiments.

[0032] Example 1

[0033] (1) A mixed solution is obtained by mixing Zn(NO3)2·6H2O, methanol and dichloromethane and a second ligand; the second ligand is 1,10-phenanthroline (CY1), 4,7-diphenyl-1,10-phenanthroline (CY2), 4,4'-dimethyl-2,2'-bipyridine (CY3), 5,5'-dimethyl-2,2-bipyridine (CY4), 4,7-dichloro-1,10-phenanthroline (CY5) or 2,2-bipyridine (CY6).

[0034] Specifically, firstly, a mixture of 1.0 mmol of Zn(NO3)2·6H2O, 2.0 mmol of methanol, and dichloromethane was added to each of six 15 cm thick-walled pressure-resistant tubes. Secondly, 1.0 mmol of the second ligand (CY1-CY6) was added to each pressure-resistant tube to obtain a mixed solution of the corresponding second ligands CY1, CY2, CY3, CY4, CY5, and CY6.

[0035] (2) Tightly cover all 6 pressure-resistant tubes and react at 65°C for 3 days.

[0036] (3) After the reaction was completed and cooled to room temperature, 2.0 mmol of the first ligand and 0.1 mL of triethylamine were added to each of the six pressure-resistant tubes; the first ligand was 5,7-dichloro-8-hydroxy-2-methylquinoline (H-ClQL).

[0037] (4) Place the pressure-resistant tube in a sealed environment and react at 80°C for 3 days. Cool to room temperature and let stand for 48 hours to obtain the target complexes CYQ1, CYQ2, CYQ3, CYQ4, CYQ5, and CYQ6 in yellow blocky crystals with a yield of 75.3%–86.4%.

[0038] In step (1), the volume ratio of methanol to dichloromethane in the mixed solution is 3:1.

[0039] The obtained complexes CYQ1–CYQ6 were identified as follows:

[0040] (1) Single crystal structure diagram of complexes CYQ1–CYQ6, as shown Figure 3-8 As shown.

[0041] (2) Infrared spectra of complexes CYQ1–CYQ6.

[0042] Specifically, the infrared spectral data of each coordination compound are as follows:

[0043] Data for complex CYQ1: Yield 79.1%; IR (KBr): 3412,3067,2924,1624,1603,1592,1565,1544,1514,1496,1454,1426,1379,1357,1278,1248,1222,1207,1193,1152,1112,1102,1025,954,938,875,865,847,826,819,775,742,729,697,654,638,593,538,499,470,450,419,405 cm⁻¹ -1 .

[0044] Data for complex CYQ2: Yield 79.1%; IR (KBr): 3403, 3059, 2923, 1618, 1603, 1547, 1521, 1496, 1428, 1391, 1358, 1277, 1250, 1233, 1195, 1149, 1112, 1092, 1032, 1001, 955, 938, 875, 856, 837, 825, 809, 774, 767, 742, 702, 666, 655, 629, 595, 576, 538, 489, 468, 453, 406 cm⁻¹ -1 .

[0045] Data for complex CYQ3: Yield 79.1%; IR (KBr): 3424, 3073, 2925, 1611, 1603, 1558, 1497, 1448, 1370, 1358, 1327, 1292, 1278, 1247, 1222, 1198, 1148, 1113, 1032, 1016, 963, 957, 919, 860, 823, 778, 742, 700, 687, 654, 601, 538, 514, 471, 412 cm⁻¹ -1 .

[0046] Data for complex CYQ4: Yield 79.1%; IR (KBr): 3420, 3113, 3022, 2921, 1603, 1563, 1546, 1496, 1480, 1449, 1432, 1389, 1356, 1311, 1277, 1249, 1233, 1193, 1165, 1150, 1112, 1042, 953, 939, 868, 829, 822, 773, 742, 733, 692, 653, 598, 538, 483, 468, 451, 419, 404 cm⁻¹ -1 .

[0047] Data for complex CYQ5: Yield 79.1%; IR (KBr): 3393,3076,3061,2987,2923,2602,1609,1568,1545,1496,1452,1432,1419,1387,1375,1355,1337,1277,1251,1216,1193,1151,1110,1084,1024,955,941,878,872,840,823,794,774,744,727,697,655,602,561,548,538,482,469,453,406 cm⁻¹ -1 .

[0048] Data for complex CYQ6: Yield 79.1%; IR (KBr): 3424, 2929, 1717, 1602, 1573, 1557, 1497, 1428, 1370, 1327, 1278, 1248, 1222, 1198, 1113, 1031, 964, 943, 860, 823, 778, 742, 701, 686, 655, 600, 539, 472, 412 cm⁻¹ -1 .

[0049] (3) Elemental analysis results are shown in Table 1.

[0050] Table 1. Elemental analysis results of compounds CYQ1-CYQ6 in the examples.

[0051]

[0052] Therefore, it can be determined that the target product of the obtained yellow blocky crystals is the complex CYQ1–CYQ6, whose structural formula is as follows: Figure 1 As shown.

[0053] To fully illustrate the application of the novel 8-hydroxyquinoline-pyridine anticancer zinc(II) complex described in this invention in pharmaceutical manufacturing, the applicant conducted in vitro and in vivo antitumor activity experiments on it.

[0054] 1. Cell lines and cell culture

[0055] This experiment used two human cell lines: drug-resistant human ovarian cancer cell line SK-OV-3 / DDP and normal HL-7702 cells.

[0056] All human cell lines were cultured in RPMI-1640 medium containing 100 U / mL penicillin, 10 wt% fetal blood, and 100 U / mL streptomycin, and incubated at 37°C in an incubator containing 5% CO2 by volume.

[0057] 2. Preparation of the test compound

[0058] All complexes and ligands used must be ≥95% pure. Their DMSO stock solutions were diluted with physiological buffer to a final solution of 20 μmol / L (final DMSO concentration ≤1%). The inhibitory effect of each compound on the growth of normal cells or selected tumor cells at this concentration was then tested.

[0059] 3. Cell growth inhibition assay (CCK-8 assay)

[0060] (1) Take cells in the logarithmic growth phase, digest them with trypsin, and prepare a cell suspension with a concentration of 5000 cells / mL using culture medium containing 10% fetal bovine serum. Seed 190.0 μL per well in a 96-well culture plate to make the cell density to be tested 1000.0 to 10000.0 wells (fill the edge wells with sterile PBS);

[0061] (2) Incubate at 37.0℃ for 24.0h with 5.0% CO2 until the cell monolayer covers the bottom of the well. Add 10μL of a certain concentration gradient of drug to each well, and set 4 replicates for each concentration gradient.

[0062] (3) Incubate at 37.0℃ for 48.0 hours with 5.0% CO2 and observe under an inverted microscope;

[0063] (4) Add 10.0 μL of WST-8 solution (5.0 mg / mL PBS, i.e. 0.5% WST-8) to each well and continue culturing for 4 h;

[0064] (5) Terminate the culture, carefully aspirate the culture medium from the wells, add 150 μL of DMSO to each well to fully dissolve the formazan precipitate, mix with a shaker, and then measure the optical density of each well using a microplate reader at a wavelength of 570.0 nm and a reference wavelength of 450.0 nm.

[0065] (6) Simultaneously set up zeroing wells (culture medium, WST-8, DMSO) and control wells (cells, WST-8, culture medium, drug dissolution medium of the same concentration, DMSO).

[0066] (7) The number of live cells can be determined based on the measured optical density value (OD value). The larger the OD value, the stronger the cell activity.

[0067] Specifically, the CCK-8 assay kit is used for rapid and sensitive detection of cell proliferation and cytotoxicity, and its inhibition rate is calculated using the following formula:

[0068]

[0069] The inhibition rate of each compound on the growth of the selected cells was calculated, and then the IC50 of each target compound on each selected cell line was calculated using the Bliss method. 50 The values ​​are shown in Table 2 below.

[0070] Table 2. IC50 of compounds CYQ1-CYQ6 on various cell lines 50 Value (μM, 48h)

[0071]

[0072] From Table 2 IC 50 Activity screening experiments showed that the complex CYQ1-CYQ6 exhibited better anticancer activity than its corresponding ligands H-ClQL, CY1-CY6, and the metal salt Zn(NO3)2·6H2O, indicating a synergistic effect between the ligands and zinc. Furthermore, CYQ2 showed the most significant inhibitory effect on the drug-resistant ovarian cancer cell line SK-OV-3 / DDP, with an IC50 concentration of [missing value]. 50 The value was 1.45±0.09μM, which was much higher than that of CYQ1, H-ClQL, CDDP, CYQ3-CYQ6, CY1-CY6, Zn(NO3)2·6H2O, and the clinical drug cisplatin (IC50). 50 (value >50 μM) and other 8-hydroxyquinoline-pyridine zinc complexes reported in the literature (IC50). 50 Value = 2.25 ± 0.13 μM); and it has very low toxicity to normal HL-7702 cells (IC50 value = 2.25 ± 0.13 μM); 50 The concentration (>50 μM) indicates that the complex CYQ1-CYQ6 exhibits tumor selectivity against SK-OV-3 / DDP tumor cells and overcomes drug resistance in clinical use, demonstrating advantages over similar zinc complexes reported in the literature. In summary, the novel 8-hydroxyquinoline-pyridine anticancer zinc(II) complex CYQ1-CYQ6 exhibits superior antitumor activity and has potential pharmaceutical value, holding promise for the preparation of various antitumor drugs.

[0073] The above are merely preferred embodiments of the present invention. It should be noted that those skilled in the art can make several modifications and improvements without departing from the structure of the present invention, and these will not affect the effectiveness of the implementation of the present invention or the practicality of the patent.

Claims

1. An anticancer zinc (II) complex of 8-hydroxyquinoline-pyridine class, characterized in that, The chemical structural formula is shown as follows: 。 2. The method of preparing 8-hydroxyquinoline-pyridine-based anticancer zinc (II) complexes according to claim 1, characterized by, The method comprises the following steps: (1) mixing a mixed solution of Zn(NO3)2·6H2O, methanol and dichloromethane and a second ligand; the second ligand is 1,10-phenanthroline, 4,7-diphenyl-1,10-phenanthroline, 4,4'-dimethyl-2,2'-bipyridine, 5 5'-dimethyl-2,2-bipyridine, 4,7-dichloro-1,10-phenanthroline or 2,2-bipyridine; (2) reacting the obtained mixed solution at 60-70°C for 3 days under sealed conditions; (3) after the reaction is completed and the temperature is cooled to room temperature, adding a first ligand and triethylamine into a pressure-resistant tube; the first ligand is 5,7-dichloro-8-hydroxy-2-methylquinoline; (4) placing the pressure-resistant tube in a sealed environment, reacting at 75-85°C for 3 days, cooling to room temperature and standing.

3. The method of preparing 8-hydroxyquinoline-pyridine-based anticancer zinc (II) complexes according to claim 2, characterized by, The mixed solution of methanol and dichloromethane is methanol: dichloromethane = 3:1 in volume.

4. The method for preparing the 8-hydroxyquinoline-pyridine anticancer zinc(II) complex according to claim 2, characterized in that, In the step (1), the amount of Zn(NO3)2·6H2O is 1.0 mmol, the amount of the mixed solution of methanol and dichloromethane is 2.0 mmol, and the amount of the second ligand is 1.0 mmol.

5. The method of preparing 8-hydroxyquinoline-pyridine-based anticancer zinc (II) complexes according to claim 2, characterized by, The reaction temperature in the step (2) is 65°C.

6. The method of preparing 8-hydroxyquinoline-pyridine-based anticancer zinc (II) complexes according to claim 2, characterized by, In the step (3), the amount of the first ligand is 2.0 mmol, and the amount of triethylamine is 0.1 mL.

7. The method of preparing 8-hydroxyquinoline-pyridine-based anticancer zinc (II) complexes according to claim 2, characterized by, The reaction temperature in the step (4) is 80°C.

8. The use of a 8-hydroxyquinoline-pyridine anticancer zinc (II) complex according to claim 1 in the preparation of a medicament for targeted therapy of drug-resistant strains of ovarian cancer, characterized in that, The drug-resistant strain is SK-OV-3 / DDP.

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