Phenanthroline calcium complex as well as preparation method and application thereof
By synthesizing o-phenanthroline calcium complex and applying it to anti-tumor drugs, the problems of nephrotoxicity and drug resistance of cisplatin drugs have been solved, achieving targeted therapeutic effects on human ovarian adenocarcinoma cells and demonstrating superior anti-tumor activity and low toxicity.
Patent Information
- Application Number
- CN202511011652.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-22
- Publication Date
- 2025-11-28
AI Technical Summary
Existing cisplatin-based drugs have issues with nephrotoxicity and drug resistance in clinical treatment, and there are no reports on the anticancer activity of o-phenanthroline calcium complex.
Three o-phenanthroline calcium complexes, [Ca2(μ2-O)2(dhg1)4(CP)2](CaL1), [Ca2(μ2-O)2(dhg2)4(CP)2](CaL2), and [Ca2(μ2-O)2(dhg3)4(CP)2](CaL3), were synthesized. These complexes were then reacted with 5-chloro-1,10-phenanthroline and Ca(NO3)2·4H2O in a CH3OH-DMF mixed solution, followed by the addition of an 8-hydroxyquinoline derivative and triethylamine, to prepare an antitumor drug with targeted therapeutic activity.
The prepared o-phenanthroline calcium complex showed significant inhibitory effects on cisplatin-resistant human ovarian adenocarcinoma cells, with IC50 values of 0.58±0.03, 0.89±0.12 and 1.93±0.26 μM, respectively, and exhibited low toxicity to normal cells, indicating potential medicinal value.
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Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of medicine, in particular to a phenanthroline calcium complex and a preparation method and application thereof. BACKGROUND
[0002] Cisplatin has insurmountable shortcomings in clinical treatment, such as nephrotoxicity and drug resistance, and therefore researchers are motivated to design new non-platinum targeted drugs. Metal calcium ions are one of the indispensable trace elements in the human body, which helps the growth of human bones and the operation of tissues and organs. The research on calcium complexes is less, but the calcium complexes basically do not have the harm of heavy metal toxicity due to the incomparable safety of calcium compared with other metals.
[0003] In addition, phenanthroline is also called 1,10-phenanthroline. A large number of literatures report that a series of phenanthroline metal complexes have good anticancer activity. However, research shows that there is no relevant report on calcium anticancer drugs and anticancer activity. SUMMARY
[0004] One of the purposes of the present application is to provide a phenanthroline calcium complex.
[0005] The phenanthroline calcium complex provided by the present application has a chemical formula of [Ca2(μ2-O)2(dhg1)4(CP)2](CaL1), [Ca2(μ2-O)2(dhg2)4(CP)2](CaL2) and [Ca2(μ2-O)2(dhg3)4(CP)2](CaL3), and a chemical structural formula as shown in the following formula:
[0006]
[0007] The second purpose of the present application is to provide a preparation method of the phenanthroline calcium complex.
[0008] The preparation method of the phenanthroline calcium complex is that 5-chloro-1,10-phenanthroline and Ca(NO3)2·4H2O are reacted in a CH3OH-DMF mixed solution at 68°C for 72 hours, then cooled to room temperature, 8-hydroxyquinoline derivatives and triethylamine are added, and reacted at 65-90°C for 72 hours, then cooled to room temperature, filtered, and the phenanthroline calcium complex is obtained.
[0009] The 8-hydroxyquinoline derivatives are 5,7-dichloro-8-hydroxyquinoline (H-dhg1), chloroiodoxyquinol (H-dhg2) or 5,7-dibromo-8-hydroxyquinoline (H-dhg3).
[0010] The synthesis route is as follows:
[0011]
[0012] Another object of the present application provides the use of the phenanthroline calcium complex.
[0013] Specifically, the use of the above-mentioned phenanthroline calcium complex in the preparation of an anti-tumor drug. The use of the above-mentioned phenanthroline calcium complex in the preparation of a targeted treatment drug for ovarian adenocarcinoma. The use of the above-mentioned phenanthroline calcium complex in the preparation of a targeted treatment drug for drug-resistant ovarian adenocarcinoma.
[0014] The present application takes 5,7-dichloro-8-hydroxyquinoline (H-dhg1), chloroiodohydroxyquinoline (H-dhg2) and 5,7-dibromo-8-hydroxyquinoline (H-dhg3) as the main ligand, 5-chloro-1,10-phenanthroline (CP) active ligand, and Ca(NO3)2·4H2O to react, synthesize three phenanthroline calcium complexes [Ca2(μ2-O)2(dhg1)4(CP)2] (CaL1), [Ca2(μ2-O)2(dhg2)4(CP)2] (CaL2) and [Ca2(μ2-O)2(dhg3)4(CP)2] (CaL3), and investigate their anticancer activity and toxicity experiments on human ovarian adenocarcinoma cisplatin-resistant cells (SK-OV-3 / DDP) and normal liver HL-7702 cells. The experimental results show that the complexes CaL1-CaL3 have a good inhibitory effect on SK-OV-3 / DDP cancer cells, and the IC 50 values are 0.58±0.03, 0.89±0.12 and 1.93±0.26 μM, respectively, which are much higher than CP, cisplatin, Ca(NO3)2·4H2O and H-dhg1–H-dhg3. It is also found that the complexes CaL1-CaL3 have little toxicity to normal HL-7702 cells; indicating that the complexes CaL1-CaL3 can target to inhibit the growth of SK-OV-3 / DDP cancer cells. In summary, the three phenanthroline calcium complexes CaL1-CaL3 show superior antitumor activity and have potential pharmaceutical value, and are expected to be used in the preparation of various anti-tumor drugs. BRIEF DESCRIPTION OF DRAWINGS
[0015] Fig. 1 X-single crystal diffraction pattern of the complex CaL1 prepared in Example 1 of the present application;
[0016] Fig. 2 X-single crystal diffraction pattern of the complex CaL2 prepared in Example 1 of the present application;
[0017] Fig. 3 X-single crystal diffraction pattern of the complex CaL3 prepared in Example 1 of the present application;
[0018] Fig. 4 Infrared spectrum of the complex CaL1 prepared in Example 1 of the present application;
[0019] Fig. 5 The infrared spectrum of the complex CaL2 obtained in Example 1 of this invention;
[0020] Fig. 6 The infrared spectrum of the complex CaL3 obtained in Example 1 of this invention is shown. Detailed Implementation
[0021] The present invention will be further described below with reference to specific embodiments, but the present invention is not limited to these embodiments.
[0022] Example 1
[0023] Dissolve 0.0215 g of 5-chloro-1,10-phenanthroline (CP, 0.1 mmol) in 1.5 mL of CH3OH-DMF mixed solution (v:v, 1.25 mL / 0.25 mL), transfer the solution to a 15.0 cm thick-walled drug-resistant tube, and then add Ca(NO3)2·4H2O (0.1 mmol, 0.0236 g) solution dissolved in 1.5 mL of CH3OH-DMF mixed solution (v:v, 1.25 mL / 0.25 mL) (perform three parallel experiments). After capping, react at 68 °C for 72 h. After the reaction was complete and cooled to room temperature, 5,7-dichloro-8-hydroxyquinoline (H-dhg1), chloroiodohydroxyquinoline (H-dhg2), and 5,7-dibromo-8-hydroxyquinoline (H-dhg3) were added to three separate tubes. Finally, 0.1 mL of triethylamine was added, the tubes were capped, and the mixture was incubated at 80°C for 72 hours. After cooling to room temperature, the mixture was filtered, and the yellow blocky crystals were washed with 10 mL of cold ethanol and ether, respectively. The crystals were then dried to obtain the o-phenanthroline calcium complexes CaL1 (yield: 54.4%), CaL2 (yield: 70.1%), and CaL3 (yield: 63.5%).
[0024] The obtained complexes CaL1-CaL3 were identified as follows:
[0025] (1) X-ray single-crystal diffraction pattern of the complex CaL1-CaL3, the spectrum is as follows: Figs. 1-3 As shown.
[0026] (2) The infrared spectrum of the complex CaL1-CaL3 is shown in the figure below. Figs. 4-6 As shown.
[0027] Data for CaL1.Yield:54.4%.IR(KBr):4030.24,3390.33,3072.44,2594.09,2202.66,1956.96,1758.43,1594.08,1555.81,1490.57,1448.50,1409.83,1395.26,1375.30,1356.88,1288.67,1249.69,1233.64,1192.18,1172.42,1140.94,1102.72,1051.37,956.18,881.30,808.75,789.76,747.62,676.81,643.56,615.01,601.28,583.31,506.26,429.56,415.88cm -1 .
[0028] Data for CaL2.Yield:70.1%.IR(KBr):4015.23,3393.07,3062.78,3039.15,1952.21,1748.80,1610.94,1589.67,1572.07,1561.05,1542.09,1510.45,1483.77,1455.43,1436.85,1420.21,1411.08,1388.90,1372.68,1340.01,1283.16,1247.22,1207.69,1191.26,1142.88,1103.16,1045.00,989.98,951.13,880.86,841.40,824.71,807.53,798.08,790.69,738.10,720.58,694.12,670.16,633.83,616.04,600.66,593.44,565.68,501.62,480.66,439.51,431.92,416.90,407.07cm -1 .
[0029] Data for CaL3.Yield:63.5%.IR(KBr):3425.87,3062.67,3040.70,2924.24,2584.30,1959.75,1661.27,1610.62,1589.73,1546 ,86,1509.36,1482.59,1455.55,1443.80,1420.18,1388.60,1373.76,1346.96,1338.30,1283.51,1264.89,1247.37,1 204.68,1191.46,1176.75,1143.50,1098.40,1045.90,992.24,948.40,933.08,882.28,873.25,852.03,824.02,807.1 7,790.74,737.23,719.49,685.43,670.01,633.15,615.65,595.36,588.29,565.23,502.47,461.31,429.21,416.75cm -1 .
[0030] (3) Elemental analysis results are shown in Table 1.
[0031] Table 1 shows the elemental analysis results of the coordination compounds CaL1-CaL3 in the examples.
[0032]
[0033] Therefore, the structural formula of the resulting complex CaL1-CaL3 can be determined as follows:
[0034]
[0035] Example 2
[0036] Dissolve 0.0215 g of 5-chloro-1,10-phenanthroline (CP, 0.1 mmol) in 1.5 mL of CH3OH-DMF mixed solution (v:v, 1.25 mL / 0.25 mL), transfer the solution to a 15.0 cm thick-walled drug-resistant tube, and then add Ca(NO3)2·4H2O (0.1 mmol, 0.0236 g) solution dissolved in 1.5 mL of CH3OH-DMF mixed solution (v:v, 1.25 mL / 0.25 mL) (perform three parallel experiments). After capping, react at 68 °C for 72 h. After the reaction was complete and cooled to room temperature, 5,7-dichloro-8-hydroxyquinoline (H-dhg1), chloroiodohydroxyquinoline (H-dhg2), and 5,7-dibromo-8-hydroxyquinoline (H-dhg3) were added to three separate tubes. Finally, 0.1 mL of triethylamine was added, the tubes were capped, and the mixture was incubated at 65°C for 72 hours. After cooling to room temperature, the mixture was filtered, and the yellow blocky crystals were washed with 10 mL of cold ethanol and ether, respectively. The crystals were then dried to obtain the o-phenanthroline calcium complexes CaL1 (yield: 37.5%), CaL2 (yield: 48.9%), and CaL3 (yield: 50.2%).
[0037] Example 3
[0038] Dissolve 0.0215 g of 5-chloro-1,10-phenanthroline (CP, 0.1 mmol) in 1.5 mL of CH3OH-DMF mixed solution (v:v, 1.25 mL / 0.25 mL), transfer the solution to a 15.0 cm thick-walled drug-resistant tube, and then add Ca(NO3)2·4H2O (0.1 mmol, 0.0236 g) solution dissolved in 1.5 mL of CH3OH-DMF mixed solution (v:v, 1.25 mL / 0.25 mL) (perform three parallel experiments). After capping, react at 68 °C for 72 h. After the reaction was complete and cooled to room temperature, 5,7-dichloro-8-hydroxyquinoline (H-dhg1), chloroiodohydroxyquinoline (H-dhg2), and 5,7-dibromo-8-hydroxyquinoline (H-dhg3) were added to three separate tubes. Finally, 0.1 mL of triethylamine was added, the tubes were capped, and the mixture was incubated at 90°C for 72 hours. After cooling to room temperature, the mixture was filtered, and the yellow blocky crystals were washed with 10 mL of cold ethanol and ether, respectively. The crystals were then dried to obtain the o-phenanthroline calcium complexes CaL1 (yield: 50.5%), CaL2 (yield: 69.3%), and CaL3 (yield: 63.0%).
[0039] To fully illustrate the pharmaceutical applications of the three o-phenanthroline calcium complexes CaL1-CaL3 described in this invention, the applicant conducted antitumor activity experiments on them.
[0040] I. Experimental Study on the Inhibitory Activity of Three phenanthroline Calcium Complexes CaL1-CaL3 on the Proliferation of Three Human Cell Lines
[0041] 1. Cell lines and cell culture
[0042] This experiment used three human cell lines: human ovarian cancer cells SK-OV-3, human ovarian adenocarcinoma cisplatin-resistant SK-OV-3 / DDP cells, and normal liver HL-7702 cells.
[0043] 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.
[0044] 2. Preparation of the test compound
[0045] All compounds used must have a purity of ≥95%. Their DMSO stock solutions were diluted with physiological buffer to a final solution of 20 μmol / L (final DMSO concentration ≤1%), and the inhibitory effect of each compound on the growth of normal cells or selected tumor cells at this concentration was tested.
[0046] 3. Cell growth inhibition experiment
[0047] The specific steps are as follows:
[0048] (1) Cells were digested, counted, and prepared to a concentration of 5×10⁻⁶. 4 Add 100 μL of cell suspension per well to each well of a 96-well cell culture plate and incubate at 37°C in a 5.0% CO2 incubator for 24.0 h.
[0049] (2) Dilute the drug to the required concentration with RPMI-1640 medium, add 100 μL of the corresponding drug-containing medium to each well of a 96-well plate, so that the final concentrations reach 0, 1.0, 2.5, 5.0, 10.0, 20.0 and 50.0 μM respectively. Set up 6 parallel experiments for each concentration. After adding the drug, place it in a 37℃, 5% CO2 incubator and continue to incubate for 48 h.
[0050] (3) Perform CCK-8 staining and measure OD values at a wavelength of λ = 450 nm: Add 10 μL of CCK-8 to each well, continue incubation in an incubator for 2 h, gently mix on a shaker for 10 min, and then measure the OD value of each well at a wavelength of λ = 450 nm using an ELISA reader. Calculate the inhibition rate of each compound on the growth of the selected cell lines, and then calculate the IC50 of each tested compound on the selected cell lines using the Bliss method. 50 Values. The results are shown in Table 2 below.
[0051] Table 2. IC50 of calcium complexes CaL1-CaL3 for various cell lines 50 Value (μM)
[0052]
[0053]
[0054] From IC 50 Based on the activity screening results, compared with human ovarian cancer cells SK-OV-3, the complex CaL1-CaL3 showed a good inhibitory effect on SK-OV-3 / DDP cancer cells, with an IC50 value of [missing information]. 50 The concentrations were 0.58±0.03, 0.89±0.12, and 1.93±0.26 μM, respectively, indicating that the activity was significantly greater than that of CP, cisplatin, Ca(NO3)2·4H2O, and H-dhg1–H-dhg3. The study also found that the complex CaL1-CaL3 exhibited low toxicity to normal HL-7702 cells, suggesting that it can target and inhibit the growth of SK-OV-3 / DDP cancer cells. In conclusion, the three o-phenanthroline calcium complexes, CaL1-CaL3, demonstrated superior antitumor activity and potential medicinal value, and are expected to be used in the preparation of various antitumor drugs.
Claims
1. α-phenanthroline calcium complexes, characterized by: Its chemical structural formula is as follows:
2. Preparation method of o-phenanthroline calcium complex: 5-chloro-1,10-phenanthroline and Ca(NO3)2·4H2O are reacted in a CH3OH-DMF mixed solution at 68℃ for 72 h. After the reaction is completed, the mixture is cooled to room temperature, and 8-hydroxyquinoline derivative and triethylamine are added. The mixture is then reacted at 65-90℃ for 72 h. After the reaction is completed, the mixture is cooled to room temperature and filtered to obtain o-phenanthroline calcium complex.
3. The method for preparing the o-phenanthroline calcium complex according to claim 1, characterized in that, The 8-hydroxyquinoline derivative is 5,7-dichloro-8-hydroxyquinoline, chloroiodohydroxyquinoline, or 5,7-dibromo-8-hydroxyquinoline.
4. The use of the o-phenanthroline calcium complex according to claim 1 in the preparation of antitumor drugs.
5. The use of the o-phenanthroline calcium complex according to claim 1 in the preparation of a targeted therapy for ovarian adenocarcinoma.
6. The use of the o-phenanthroline calcium complex according to claim 1 in the preparation of targeted therapy for drug-resistant ovarian adenocarcinoma.