8-hydroxyquinoline-based anticancer zinc(ii) complexes, methods for their synthesis and use

Abstract

The application discloses an 8-hydroxyquinoline anticancer zinc (II) complex and a synthesis method and application thereof. The 8-hydroxyquinoline anticancer zinc (II) complex is synthesized by taking 5,7-diiodo-8-hydroxyquinoline, 5,7-dichloro-8-hydroxyquinoline, 5,7-dibromo-8-hydroxyquinoline and 2-methyl-8-hydroxyquinoline as a first ligand and taking 1,10-phenanthroline, 4,7-diphenyl-1,10-phenanthroline, 4,7-dichloro-1,10-phenanthroline, 5,6-dimethyl-1,10-phenanthroline and 5-chloro-1,10-phenanthroline as auxiliary ligands. The 8-hydroxyquinoline anticancer zinc (II) complex has good inhibiting effects on SK-OV-3CR cells of an ovarian cancer drug-resistant strain, is higher than that of cisplatin drugs, overcomes drug resistance of clinical drugs, has potential medicinal value and can be used for preparation of various antitumor drugs.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medicine, in particular to 8-hydroxyquinoline anticancer zinc (II) complexes and a synthesis method and application thereof. BACKGROUND

[0002] Cancer is generally treated by radiotherapy, surgery, chemotherapy and other methods, but only surgery and radiotherapy are important means of local treatment, however, platinum drugs in chemotherapy, while inhibiting the growth of tumor cells, also have different degrees of toxicity to normal cells in the body. Therefore, in recent years, the development and treatment of targeted non-platinum drugs show a promising prospect (Guo, Z.; et al. Chem. Soc. Rev., 2013, 42: 202-224.), but it is still in the clinical trial stage, and further observation and accumulation are needed for clinical use, so there is still a big gap from large-scale clinical use.

[0003] At present, the research on the anticancer activity of 8-hydroxyquinoline zinc complexes against SK-OV-3 / DDP is still blank, and the synthesis of zinc complexes and the research on their anticancer activity using 8-hydroxyquinoline (H-Q1-H-Q4) as the first ligand and 1,10-phenanthroline derivatives (D1-D5) as the auxiliary ligand have not been reported in the literature. SUMMARY

[0004] One of the purposes of the present application is to provide 8-hydroxyquinoline anticancer zinc (II) complexes.

[0005] Specifically, the present application synthesizes zinc complexes using 8-hydroxyquinoline (H-Q1-H-Q4) as the first ligand and 1,10-phenanthroline derivatives (D1-D5) as the auxiliary ligand.

[0006] The application synthesizes 8-hydroxyquinoline anticancer zinc (II) complexes with chemical formula [Zn(Q1)2(D1)]·CH3OH (DQ1), [Zn(Q1)2(D2)] (DQ2), [Zn(Q2)2(D2)] (DQ3), [Zn(Q3)2(D2)] (DQ4), [Zn(Q3)2(D1)] (DQ5), [Zn(Q3)2(D3)]·CH3OH (DQ6), [Zn(Q3)2(D4)]·CH2Cl2 (DQ7), [Zn(Q3)2(D5)]·CH2Cl2 (DQ8) and [Zn(Q4)2(D1)]·CH3OH (DQ9) by using 5,7-diiodo-8-hydroxyquinoline (H-Q1), 5,7-dichloro-8-hydroxyquinoline (H-Q2), 5,7-dibromo-8-hydroxyquinoline (H-Q3) and 2-methyl-8-hydroxyquinoline (H-Q4) as the first ligand, and 1,10-phenanthroline (D1), 4,7-diphenyl-1,10-phenanthroline (D2), 4,7-dichloro-1,10-phenanthroline (D3), 5,6-dimethyl-1,10-phenanthroline (D4) and 5-chloro-1,10-phenanthroline (D5) as the auxiliary ligand, and the chemical structural formula is shown in the following formula:

[0007]

[0008] The second object of the application is to provide a synthesis method of the above-mentioned 8-hydroxyquinoline anticancer zinc (II) complex, which comprises the following steps: weighing 0.1 mmol of the auxiliary ligand and 0.1 mmol of the metal salt Zn(NO3)2·6H2O in a 15.0 cm long thick-walled drug-resistant tube, then adding 3.5 mL of a mixed solution of MeOH and CH2Cl2 with a volume ratio of 6:1, covering the lid, and performing a coordination reaction at 65 ℃ for 3 days; after the reaction is completed, cooling to room temperature, opening the lid of the system, then adding 0.2 mmol of the first ligand 8-hydroxyquinoline derivative and 0.1 mL of triethylamine, covering the lid again, and performing a reaction at 80 ℃ for 3 days; and after cooling and standing for 48 h, obtaining the target product in the form of yellow block crystals.

[0009] The 8-hydroxyquinoline derivative is 5,7-diiodo-8-hydroxyquinoline (H-Q1), 5,7-dichloro-8-hydroxyquinoline (H-Q2), 5,7-dibromo-8-hydroxyquinoline (H-Q3) or 2-methyl-8-hydroxyquinoline (H-Q4).

[0010] The auxiliary ligand is 1,10-phenanthroline (D1), 4,7-diphenyl-1,10-phenanthroline (D2), 4,7-dichloro-1,10-phenanthroline (D3), 5,6-dimethyl-1,10-phenanthroline (D4) or 5-chloro-1,10-phenanthroline (D5).

[0011] The synthetic route of the present application is as follows:

[0012]

[0013] Notably:

[0014] (1) The triethylamine should be 0.1 mL, if less than this amount, the overall reaction yield is very low, if more than this amount, a large amount of powder precipitates 2, leading to incomplete reaction.

[0015] (2) All the reaction temperatures have been explored, only the given temperature is suitable to obtain the target product of yellow block crystal, and other temperature reaction yield is low or the reaction is paste.

[0016] (3) MeOH and CH2Cl2 must be in the same volume ratio v:v = 6:1, the reaction product has less impurities, and it is easy to separate.

[0017] Another object of the present application provides the application of the above-mentioned 8-hydroxyquinoline anticancer zinc (II) complex.

[0018] Specifically, the application of the 8-hydroxyquinoline anticancer zinc (II) complex in the preparation of a drug for targeted treatment of ovarian cancer. Further relates to the application of the 8-hydroxyquinoline anticancer zinc (II) complex in the preparation of a drug for targeted treatment of drug-resistant ovarian cancer.

[0019] The present application takes 5,7-diiodo-8-hydroxyquinoline (H-Q1), 5,7-dichloro-8-hydroxyquinoline (H-Q2), 5,7-dibromo-8-hydroxyquinoline (H-Q3) and 2-methyl-8-hydroxyquinoline (H-Q4) as the first ligand, and 1,10-phenanthroline (D1), 4,7-diphenyl-1,10-phenanthroline (D2), 4,7-dichloro-1,10-phenanthroline (D3), 5,6-dimethyl-1,10-phenanthroline (D4) and 5-chloro-1,10-phenanthroline (D5) as auxiliary ligands to synthesize new 8-hydroxyquinoline anticancer zinc (II) complexes DQ1-DQ9, and their activities and toxicity experiments on human ovarian cancer drug-resistant cell line SK-OV-3 / DDP (abbreviation: SK-OV-3CR, same below) and normal HL-7702 cells are investigated. The experimental results show that DQ3 has the most obvious inhibitory effect on ovarian cancer drug-resistant cell line SK-OV-3CR, IC 50The value is 2.25±0.13 μM, much larger than DQ1-DQ2, DQ4-DQ9, H-Q1-H-Q4, D1-D5, metal salt Zn(NO3)2·6H2O and clinical drug cisplatin, and the toxicity to normal HL-7702 cells is very small, which shows that the complex has selectivity to tumor cell SK-OV-3CR, and is superior to the complexes reported in the literature. In addition, the new 8-hydroxyquinoline anticancer zinc (II) complex DQ1-DQ9 has good inhibition effect on ovarian cancer drug-resistant cell line SK-OV-3CR, which is higher than that of cisplatin drug, and overcomes the drug resistance of clinical drug. In summary, the new 8-hydroxyquinoline anticancer zinc (II) complex DQ1-DQ9 shows superior antitumor activity, has potential medicinal value, and is expected to be used for the preparation of various antitumor drugs. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 The X-ray single crystal structure diagram of the complex DQ1 prepared in the embodiment 1 of the application;

[0021] Figure 2 The X-ray single crystal structure diagram of the complex DQ2 prepared in the embodiment 1 of the application;

[0022] Figure 3 The X-ray single crystal structure diagram of the complex DQ3 prepared in the embodiment 1 of the application;

[0023] Figure 4 The X-ray single crystal structure diagram of the complex DQ4 prepared in the embodiment 1 of the application;

[0024] Figure 5 The X-ray single crystal structure diagram of the complex DQ5 prepared in the embodiment 1 of the application;

[0025] Figure 6 The X-ray single crystal structure diagram of the complex DQ6 prepared in the embodiment 1 of the application;

[0026] Figure 7 The X-ray single crystal structure diagram of the complex DQ7 prepared in the embodiment 1 of the application;

[0027] Figure 8 The X-ray single crystal structure diagram of the complex DQ8 prepared in the embodiment 1 of the application;

[0028] Figure 9 The X-ray single crystal structure diagram of the complex DQ9 prepared in the embodiment 1 of the application;

[0029] Figure 10 The infrared spectrum of the complex DQ1 prepared in the embodiment 1 of the application;

[0030] Figure 11The infrared spectrum of complex DQ2 obtained in Example 1 of this invention;

[0031] Figure 12 The infrared spectrum of complex DQ3 obtained in Example 1 of this invention;

[0032] Figure 13 The infrared spectrum of complex DQ4 obtained in Example 1 of this invention;

[0033] Figure 14 The infrared spectrum of complex DQ5 obtained in Example 1 of this invention;

[0034] Figure 15 The infrared spectrum of complex DQ6 obtained in Example 1 of this invention;

[0035] Figure 16 The infrared spectrum of complex DQ7 obtained in Example 1 of this invention;

[0036] Figure 17 The infrared spectrum of complex DQ8 obtained in Example 1 of this invention;

[0037] Figure 18 The infrared spectrum of complex DQ9 obtained in Example 1 of this invention is shown. Detailed Implementation

[0038] The present invention will be further described below with reference to specific embodiments, but the present invention is not limited to these embodiments.

[0039] Example 1

[0040] In a 15.0 cm long thick-walled drug-resistant tube, weigh out 0.1 mmol of 1,10-phenanthroline (D1), 4,7-diphenyl-1,10-phenanthroline (D2), 4,7-dichloro-1,10-phenanthroline (D3), 5,6-dimethyl-1,10-phenanthroline (D4) or 5-chloro-1,10-phenanthroline (D5), and 0.1 mmol of the metal salt Zn(NO3)2·6H2O, respectively, and then add 3.5 mL of the solution. A mixed solution of MeOH and CH2Cl2 (v:v = 6:1) was prepared, capped, and subjected to a coordination reaction at 65°C for 3 days. After the reaction, the mixture was cooled to room temperature, the cap was opened, and 0.2 mmol of 5,7-diiodo-8-hydroxyquinoline (H-Q1), 5,7-dichloro-8-hydroxyquinoline (H-Q2), 5,7-dibromo-8-hydroxyquinoline (H-Q3), or 2-methyl-8-hydroxyquinoline (H-Q4) and 0.1 mL of triethylamine were added, respectively. The cap was closed again, and the mixture was subjected to a reaction at 80°C for 3 days. After cooling and standing for 48 hours, the target products DQ1–DQ9, which are yellow blocky crystals, were obtained. The yields were 69.9%–94.4%.

[0041] The resulting product was identified:

[0042] (1) X-ray single crystal structure of compound DQ1-DQ9, the spectrum of which is shown in Figure 1 -9.

[0043] (2) Infrared spectrum of compound DQ1-DQ9, the spectrum of which is shown in Figure 10 -18.

[0044] Data for DQ1.Yield:94.4%.IR(KBr):3352,3056,2928,2813,2548,1760,1675,1623,1587,1578,1553,1533,1517,1472,1444,1387,1379,1347,1277,1250,1203,1136,1103,1045,1034,927,882,866,859,842,806,788,738,727,670,652,636,590,558cm -1 .

[0045] Data for DQ2.Yield:90.5%.IR(KBr):3401,3052,2550,1930,1759,1676,1614,1592,1556,1535,1493,1475,1444,1428,1388,1348,1279,1251,1203,1157,1135,1108,1091,1046,1033,1019,1000,927,880,840,804,788,764,739,700,669,652,628,594,575,560,546cm -1 .

[0046] Data for DQ3.Yield:69.9%.IR(KBr):3380,3097,3064,3030,2966,2828,2606,1615,1593,1556,1521,1489,1452,1427,1393,1381,1356,1280,1252,1232,1219,1198,1158,1138,1113,1092,1048,1033,1020,1000,960,880,859,851,812,792,766,743,701,675,665,649,628,595,584,575,546cm -1 .

[0047] Data for DQ4. Yield: 89.9%. IR (KBr): 3392, 3059, 2598, 1971, 1927, 1749, 1614, 1592, 1547, 1520, 1483, 1454, 1427, 1378, 1394, 1352, 1280, 1251, 1232, 1210, 1182, 1157, 1135, 1109, 1092, 1074, 1047, 1032, 1020, 1000, 964, 941, 875, 855, 833, 812, 791, 766, 742, 701, 681, 671, 642, 628, 594, 583, 573, 545 cm -1 .

[0048] Data for DQ5. Yield: 78.1%. IR (KBr): 3068, 2940, 2824, 1956, 1739, 1624, 1592, 1556, 1517, 1484, 1445, 1390, 1376, 1361, 1281, 1251, 1237, 1219, 1137, 1108, 1050, 1030, 948, 881, 874, 861, 806, 787, 737, 692, 667, 641, 592, 576, 505 cm -1 .

[0049] Data for DQ6. Yield: 80.9%. IR (KBr): 3674, 3366, 3075, 2941, 2809, 2597, 2192, 1938, 1750, 1615, 1596, 1575, 1569, 1549, 1511, 1484, 1450, 1418, 1389, 1376, 1352, 1312, 1290, 1250, 1236, 1213, 1174, 1138, 1130, 1107, 1083, 1032, 941, 874, 858, 832, 807, 788, 742, 685, 667, 642, 592, 572, 547, 519 cm -1 .

[0050] Data for DQ7. Yield: 91.7%. IR (KBr): 3787, 3402, 3069, 3045, 2938, 1958, 1740, 1605, 1550, 1520, 1484, 1450, 1392, 1378, 1352, 1311, 1250, 1213, 1166, 1139, 1108, 1072, 1051, 1031, 942, 873, 859, 806, 788, 741, 735, 688, 670, 642, 620, 593, 573, 547 cm -1 .

[0051] Data for DQ8. Yield: 80.4%. IR (KBr): 3401, 3067, 1739, 1611, 1586, 1554, 1520, 1484, 1452, 1384, 1360, 1252, 1219, 1138, 1110, 1051, 1032, 948, 890, 874, 862, 807, 787, 745, 733, 691, 667, 644, 593, 576, 537 cm -1 .

[0052] Data for DQ9. Yield: 92.0%. IR (KBr): 3664, 3273, 3164, 3053, 1621, 1608, 1585, 1559, 1516, 1504, 1459, 1425, 1369, 1340, 1369, 1298, 1285, 1236, 1221, 1208, 1174, 1141, 1102, 1062, 1034, 964, 916, 866, 847, 832, 796, 777, 740, 729, 723, 637, 603, 558, 517, 508 cm -1 .

[0053] (3) Elemental analysis results, as shown in Table 1.

[0054] Table 1 Elemental analysis results of compounds DQ1-DQ9 in the examples

[0055]

[0056] Therefore, the target products DQ1-DQ9 of the obtained yellow block crystals can be determined, and the structural formulas are as follows:

[0057]

[0058] In order to fully illustrate the use of the nine novel 8-hydroxyquinoline anticancer zinc (II) complexes DQ1-DQ9 in the preparation of medicines, the applicant has carried out in vitro and in vivo anti-tumor activity experiments.

[0059] One, the proliferation inhibition activity experiment of the novel 8-hydroxyquinoline anticancer zinc (II) complexes DQ1-DQ9 on two human cell lines

[0060] 1. Cell lines and cell culture

[0061] In this experiment, two human cell lines, human ovarian cancer drug-resistant cell line SK-OV-3CR and normal HL-7702 cells, were selected.

[0062] All human cell lines were cultured in RPMI-1640 culture medium containing 100 U / mL penicillin, 10 wt% calf blood, and 100 U / mL streptomycin, and incubated in a 37°C incubator containing 5% CO2 by volume.

[0063] 2. Preparation of test compounds

[0064] All compounds used in this experiment had a purity of ≥95%. The DMSO stock solution of each compound was diluted with a physiological buffer to a final concentration of 20 μmol / L (the final concentration of DMSO was ≤1%), and the degree of inhibition of the growth of normal cells or selected tumor cells at this concentration was tested.

[0065] 3. Cell growth inhibition experiment (MTT method)

[0066] (1) Take the normal cells or tumor 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% calf serum. Inoculate 190 μL of the cell suspension into each well of a 96-well culture plate to achieve a cell density of 1000-10000 wells (the edge wells are filled with sterile PBS);

[0067] (2) Incubate at 37°C for 24 h in a 5% CO2 incubator until the cell monolayer covers the bottom of the well. Add 10 μL of the drug at a certain concentration gradient to each well, and set 4 replicate wells for each concentration gradient;

[0068] (3) Incubate at 37°C for 48 h under a 5% CO2 incubator, and observe under an inverted microscope;

[0069] (4) Add 10 μL of MTT solution (5 mg / mL PBS, i.e. 0.5% MTT) to each well, and continue to culture for 4 h;

[0070] (5) Terminate the culture, carefully suck the culture solution in the hole, add 150 μL of DMSO to each hole to dissolve the methaemoglobin precipitate thoroughly, shake to mix, then measure the optical density value of each hole in an enzyme marker at a wavelength of 570 nm and a reference wavelength of 450 nm;

[0071] (6) Set the zero hole (culture medium, MTT, DMSO) and the control hole (cells, culture solution, MTT, the same concentration of drug dissolving medium, DMSO) at the same time.

[0072] (7) According to the measured optical density value (OD value), determine the number of living cells, the greater the OD value, the stronger the cell activity. Use the formula:

[0073]

[0074] Calculate the inhibition rate of each compound on the growth of the selected cells, and then calculate the IC 50 value of each tested compound on the selected cells of each cell strain respectively by Bliss method. The results are shown in Tables 2 and 3 below.

[0075] Table 2. IC 50 values (μM, 24h) of compounds DQ1-DQ9 on various cell strains

[0076]

[0077] Table 3. IC 50 values (μM, 24h) of compounds on various cell strains

[0078]

[0079] From the IC 50 activity screening results in Tables 2 and 3, DQ3 has the most obvious inhibitory effect on the ovarian cancer drug-resistant strain cell SK-OV-3CR, with an IC 50 value of 2.25±0.13 μM, which is much greater than DQ1-DQ2, DQ / 4-DQ9, H-Q1-H-Q4, D1-D5, metal salt Zn(NO3)2·6H2O and the clinical drug cisplatin, and has very little toxicity to normal HL-7702 cells, indicating that the complex has selectivity for tumor cells SK-OV-3CR; while the research group of Chen Zhenfeng reported that the four 8-hydroxyquinoline derivative anti-cancer zinc complexes have very high anti-cancer activity (IC 50 = 44±1 nM) but have great toxicity to normal cells HL-7702 (IC 50The value is lower than 1.78 ± 0.05 μM (Chen, Z.-F..; et al. Eur. J. Med. Chem., 2013, 69: 554-563.), which has no tumor selectivity and great toxicity, so the reported complex selectivity is better than the literature reported, the normal cell toxicity is small, the selectivity is high, and it is more advantageous. In addition, the new 8-hydroxyquinoline anticancer zinc (II) complex DQ1-DQ9 has good inhibition effect on ovarian cancer drug-resistant cell line SK-OV-3CR, which is higher than that of cisplatin drug (60.37 ± 1.14 μM), overcoming the drug resistance of clinical drugs. In summary, the new 8-hydroxyquinoline anticancer zinc (II) complex DQ1-DQ9 shows superior antitumor activity, has potential pharmaceutical value, and is expected to be used for the preparation of various antitumor drugs.

Claims

1. An anticancer zinc (II) complex of 8-hydroxyquinoline characterized in that, The chemical structural formula is shown in the following formula: 。 2. The method of claim 1, wherein the 8-hydroxyquinoline-based zinc complex is synthesized by reacting zinc chloride with 8-hydroxyquinoline in a solvent. 15.0 cm long thick-walled drug-resistant tube, respectively, 0.1 mmol of auxiliary ligand and 0.1 mmol of metal salt Zn(NO3)2·6H2O, followed by 3.5 mL of MeOH and CH2Cl2 mixed solution with a volume ratio of 6:1, cover the lid, the coordination reaction at 65℃ for 3 days, after the reaction is cooled to room temperature, open the cover of the system, then respectively 0.2 mmol of the first ligand 8-hydroxyquinoline derivatives and 0.1 mL triethylamine, cover the lid again, at 80℃ for 3 days, after cooling and standing for 48h, the target product of yellow block crystal is obtained; The 8-hydroxyquinoline derivative is 5,7-diiodo-8-hydroxyquinoline, 5,7-dichloro-8-hydroxyquinoline, 5,7-dibromo-8-hydroxyquinoline or 2-methyl-8-hydroxyquinoline; The auxiliary ligand is 1,10-phenanthroline, 4,7-diphenyl-1,10-phenanthroline, 4,7-dichloro-1,10-phenanthroline, 5,6-dimethyl-1,10-phenanthroline or 5-chloro-1,10-phenanthroline.

3. The use of the 8-hydroxyquinoline anticancer zinc (II) complex of claim 1 in the preparation of a drug for targeted treatment of drug-resistant strains of ovarian cancer, characterized in that, The drug-resistant strain is SK-OV-3CR.

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