8-hydroxyl dictamnine iridium (III) complex as well as synthesis method and application thereof

Abstract

The invention discloses an 8-hydroxyl dictamnine iridium (III) complex as well as a synthesis method and application thereof. The novel 8-hydroxyl dictamnine iridium (III) complex with high activity in vitro and vivo is synthesized by taking an 8-hydroxyl dictamnine derivative as an active ligand and reacting with a 2-phenylpyridine iridium (III) dimer or a derivative thereof as a metal salt. The 8-hydroxyl dictamnine iridium (III) complex has a good inhibition effect on human ovarian cancer cells SK-OV-3 and cis-platinum drug-resistant strains SK-OV-3 / DDP, shows excellent in-vivo and in-vitro antitumor activity, has potential medicinal value, and is expected to be used for preparing various antitumor drugs.

Description

Technical Field

[0001] The invention relates to the technical field of medicines, and in particular to an 8-hydroxydandridine iridium (III) complex and a synthesis method and application thereof. Background Art

[0002] Cisplatin drugs have achieved very good therapeutic effects in the clinical treatment of cancer, but the toxicity and drug resistance they produce cannot be ignored, which limits their wide clinical application (Guo, Z.; et al. Chem. Soc. Rev., 2013, 42: 202-224.), so there is an urgent need to develop a class of targeted, low-toxic non-platinum anticancer drugs.

[0003] Literature reports show that 8-hydroxyquinoline derivative iridium complexes have good anticancer activity against tumor cells, especially HeLa cancer cells, IC 50 The value is as low as 0.05±0.04μM (Liang, F.-P., et al. Eur. J. Med. Chem., 2020, 192, 112192.); however, the synthesis and anticancer effect research of 8-hydroxydandruff metal complexes is still blank. Summary of the invention

[0004] One of the objects of the present invention is to provide an 8-hydroxy-dandrogenine iridium (III) complex.

[0005] Specifically, the present invention uses 8-hydroxyquinoline and its derivatives (8-hydroxyquinoline (H-Rob1) and 8-hydroxyquinoline (H-Rob2)) as ligands, and 2-phenylpyridine or 3-methyl-2-phenylpyridine as auxiliary ligands to synthesize 8-hydroxyquinoline iridium (III) complexes, including [Ir(Rob1)(PPY1) 2 ]·CH 3 OH(Ro1), [Ir(Rob1)(PPY2) 2 ]·CH 3 OH(Ro2), [Ir(Rob2)(PPY2) 2 ](Ro3) and [Ir(Rob2)(PPY2) 2 ](Ro4), whose chemical structure is shown below:

[0006]

[0007] The second object of the present invention is to provide a method for synthesizing the above-mentioned 8-hydroxyquinoline iridium (III) complex: weigh 2-phenylpyridine iridium (III) dimer (H-PPY1 dimer) or its derivatives and place them in a drug-resistant tube, add 8-hydroxyquinoline or its derivatives, then add a solvent, cover the lid, react at 45°C for 2 days, react at 65°C for 1 day, and finally react at 80°C for 1 day. After the reaction is completed, cool it to 25°C to obtain the obtained product.

[0008] The solvent was 2.0 mL MeOH and 0.1 mL DMSO or 2.0 mL MeOH and 0.1 mL CH 2 Cl 2 .

[0009] The 8-hydroxyquinoline derivatives are 8-hydroxyrobinine (H-Rob1) and 8-hydroxyquinoline (H-Rob2).

[0010] The 2-phenylpyridine iridium (III) dimer derivative is 3-methyl-2-phenylpyridine iridium (III) dimer (H-PPY2 dimer).

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

[0012]

[0013] In the present invention,

[0014] (a) Synthesis conditions of compounds Ro1 and Ro3: solvent was 2.0 mL MeOH + 0.1 mL DMSO, reaction was carried out at 45°C for 2 days, at 65°C for 1 day, and finally at 80°C for 1 day.

[0015] (b) Synthesis conditions of compounds Ro2 and Ro4: Solvent: 2.0 mL MeOH + 0.1 mL CH 2 Cl 2 , react at 45°C for 2 days, react at 65°C for 1 day, and finally react at 80°C for 1 day.

[0016] Another object of the present invention is to provide the application of the above 8-hydroxy-dandrogenine iridium (III) complex.

[0017] Specifically, the invention relates to the use of 8-hydroxydandridine iridium (III) complex in the preparation of drugs for targeted treatment of ovarian cancer, and further relates to the use of 8-hydroxydandridine iridium (III) complex in the preparation of drugs for targeted treatment of drug-resistant ovarian cancer.

[0018] Compared with the prior art, the present invention uses 8-hydroxybactamine derivatives H-Rob1 and H-Rob2 as active ligands, and reacts with 2-phenylpyridine iridium (III) dimer (H-PPY1 dimer) and 3-methyl-2-phenylpyridine iridium (III) dimer (H-PPY2 dimer) as metal salts to synthesize four novel 8-hydroxybactamine iridium (III) complexes Ro1-Ro4 with high in vitro and in vivo activity, and investigates their activity and toxicity experiments on human ovarian cancer cells SK-OV-3, cisplatin-resistant strain SK-OV-3 / DDP and normal HL-7702 liver cells. The experimental results show that complex Ro2 has a good anticancer effect on SK-OV-3 and SK-OV-3 / DDP cancer cells, and its IC 50 The values ​​were 6.39±0.80μM and 4.03±0.19μM, especially for SK-OV-3 / DDP cancer cells, and their activity was much greater than that of H-Rob1 (30.27±0.99μM), H-Rob2 (36.11±0.23μM), Ro1 (7.08±0.50μM), Ro3 (13.82±1.04μM), Ro4 (10.16±0..31μM) and cisplatin (>50μM); and Ro1-Ro4 had little inhibitory effect on normal HL-7702 cells (IC 50 Value>50μM); it shows that complexes Ro1-Ro4 can overcome cisplatin resistance and target the proliferation of SK-OV-3 / DDP cancer cells. More importantly, in vivo tumor inhibition experiments show that complex Ro2 has a good tumor inhibition effect on the nude mouse model of human ovarian cancer cells SK-OV-3, with an inhibition rate of up to 60.4%. In short, the new 8-hydroxy-dandridine iridium (III) complexes Ro1-Ro4 show excellent in vitro and in vivo anti-tumor activity, have potential medicinal value, and are expected to be used in the preparation of various anti-tumor drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 is the X-ray single crystal diffraction pattern of the complex Ro1 prepared in Example 1 of the present invention;

[0020] Figure 2 This is the X-ray single crystal diffraction pattern of the complex Ro2 prepared in Example 1 of the present invention;

[0021] Figure 3 is the hydrogen nuclear magnetic resonance spectrum of the complex Ro1 prepared in Example 1 of the present invention;

[0022] Figure 4 is the hydrogen nuclear magnetic resonance spectrum of the complex Ro2 prepared in Example 1 of the present invention;

[0023] Figure 5 is the hydrogen nuclear magnetic resonance spectrum of the complex Ro3 prepared in Example 1 of the present invention;

[0024] Figure 6 This is the carbon NMR spectrum of the complex Ro3 prepared in Example 1 of the present invention;

[0025] Figure 7 This is the hydrogen nuclear magnetic resonance spectrum of the complex Ro4 prepared in Example 1 of the present invention;

[0026] Figure 8 This is the carbon NMR spectrum of the complex Ro4 prepared in Example 1 of the present invention. DETAILED DESCRIPTION

[0027] The present invention will be further described below with reference to specific examples, but the present invention is not limited to these examples.

[0028] Example 1

[0029] (1) 2-phenylpyridine iridium (III) dimer (H-PPY1 dimer) and 3-methyl-2-phenylpyridine iridium (III) dimer (H-PPY2 dimer) were synthesized according to the synthesis method of the metal salt reaction (Chao, H., et al. Biomaterials, 2015, 63, 128-136).

[0030] (2) Synthetic complex

[0031] Weigh 0.1 mmol of H-PPY1 dimer and place it in a 15.0 cm thick-walled drug-resistant tube, weigh 0.1 mmol of H-Rob1, then add 2.0 mL of MeOH and 0.1 mL of DMSO. After covering the lid of the drug-resistant bottle, the system is reacted at 45°C for 2 days, at 65°C for 1 day, and finally at 80°C for 1 day. After the reaction is completed, cool it to 25°C, open the lid of the system, and then obtain the yellow product Ro1.

[0032] Weigh 0.1 mmol of H-PPY1 dimer and place it in a 15.0 cm thick-walled drug-resistant tube. Weigh 0.1 mmol of H-Rob1 and then add 2.0 mL of MeOH and 0.1 mL of CH 2 Cl 2 After covering the lid of the drug-resistant bottle, the system was reacted at 45°C for 2 days, at 65°C for 1 day, and finally at 80°C for 1 day. After the reaction was completed, it was cooled to 25°C, the lid of the system was opened, and then the yellow product Ro2 was obtained.

[0033] Weigh 0.1 mmol H-PPY2 dimer and place it in a 15.0 cm thick-walled drug-resistant tube, weigh 0.1 mmol H-Rob2, then add 2.0 mL MeOH and 0.1 mL DMSO. After covering the lid of the drug-resistant bottle, the system is reacted at 45°C for 2 days, at 65°C for 1 day, and finally at 80°C for 1 day. After the reaction is completed, cool it to 25°C, open the lid of the system, and then obtain the yellow product Ro3.

[0034] Weigh 0.1 mmol H-PPY2 dimer and place it in a 15.0 cm thick-walled drug-resistant tube. Weigh 0.1 mmol H-Rob2 and then add 2.0 mL MeOH and 0.1 mL CH 2 Cl 2 After covering the lid of the drug-resistant bottle, the system was reacted at 45°C for 2 days, at 65°C for 1 day, and finally at 80°C for 1 day. After the reaction, it was cooled to 25°C, the lid of the system was opened, and then the yellow product Ro4 was obtained.

[0035] The obtained products Ro1-Ro4 were identified:

[0036] (1) X-ray single crystal diffraction patterns of complexes Ro1 and Ro2, as shown in the following figure Figure 1 and 2 shown.

[0037] (2) The hydrogen and carbon nuclear magnetic resonance spectra of the complexes Ro1-Ro4 are shown in the following figure. Figure 3-8 shown.

[0038] Data for Ro1.Yield:49.5%. 1 H NMR (600 MHz, DMSO-d 6 )δ9.81(d,J=5.3Hz,1H),9.53(d,

[0039] J=5.0Hz,1H),8.26(d,J=8.0Hz,1H),8.17(d,J=7.9Hz,1H),8.12-8.07(m,1H),8.03-7.99(m,1H),7.78(d,J=6.9Hz,1H),7.75(dd,J=11.8,5.0Hz,2H),7.72-7.69(m,2H),7.59-7.55(m,1H),7.47-7.43(m,1H),7.33(dd,J=9.3,2.7Hz,1H),6.89(dd,J=10.8,4.1Hz,1H),6.84(t,J=7.4Hz,1H),6.78-6.74(m,1H),6.69(dd,J=11.7,4.3Hz,2H),6.25(d,J=7.1Hz,1H),5.66(d,J=7.2Hz,1H),4.39(s,3H).

[0040] Data for Ro2.Yield:68.4%. 1 H NMR(600MHz,DMSO-d 6 )δ9.85(d,J=5.1Hz,1H),9.67(d,

[0041] J=5.2Hz,1H),7.96(d,J=7.5Hz,1H),7.92-7.86(m,3H),7.84(m,3H),7.51-7.44(m,1H),7.39-7.31(m,1H),6.90(t,J=7.4Hz,2H),6.84(t,J=7.4Hz,2H),6.74(t,J=7.3Hz,1H),6.66(t,J=7.3Hz,1H),6.24(d,J=7.5Hz,1H),5.61(d,J=7.5Hz,1H),4.38(s,3H),2.80(s,3H),2.77(s,3H).

[0042] Data for Ro3.Yield:75.1%. 1 H NMR(600MHz,DMSO-d 6 )δ9.81(d,J=5.5Hz,1H),9.54(d,

[0043] J=5.1Hz,1H),8.26(d,J=8.1Hz,2H),8.18(d,J=8.2Hz,2H),8.09(t,J=7.8Hz,1H),8.01(t,J=7.8Hz,1H),7.78(d,J=7.8Hz,2H),7.73(d,J=7.2Hz,2H),7.57(t,J=6.6Hz,1H),7.45(t,J=6.6Hz,1H),6.89(t,J=7.5Hz,1H),6.84(t,J=7.5Hz,1H),6.76(t,J=7.5Hz,2H),6.69(t,J=7.4Hz,2H),6.25(d,J=7.3Hz,1H),5.66(d,J=7.3Hz,1H). 13 CNMR(151MHz,DMSO-d 6 )Δ167.34,166.96,152.09,152.00,150.63,145.37,143.75,143.16,139.18,138.11,131.24,129.89,129.62,128.93,124.77,123.77,123.52,122.78,122.23,121.94,120.04,119.41.

[0044] Data for Ro4.Yield:55.4%. 1 H NMR(600MHz,DMSO-d 6 )δ9.85(d,J=4.7Hz,1H),9.67(d,

[0045] J=5.1Hz,1H),7.96(d,J=7.5Hz,1H),7.92(t,J=8.1Hz,1H),7.87(m,2H),7.83(d,J=8.0Hz,2H),7.58(m,2H),7.35(dd,J=8.0,5.2Hz,2H),6.90(m,1H),6.85(m,2H),6.75(m,2H),6.67(dd,J=7.0,2.3Hz,2H),6.25(d,J=7..5Hz,1H),5.62(d,J=7.5Hz,1H),2.80(s,5H),2.78(s,5H). 13 C NMR(151MHz,DMSO-d 6)Δ169.08,165.20,164.71,150.73,,149.46,146.21,146.05,145.27 ,144.95,143.38,142.75,141.97,141.29,137.00,132.54,131.49,,1 31.26,130.31,129.46,128.89,128.24,127.92,127.31,122.70,122 .11,121.92,121.77,121.60,,120.30,120.15,116.09,22.93,22.44.

[0046] Therefore, the obtained products Ro1-Ro4 were determined, and their structural formulas are as follows:

[0047]

[0048] In order to fully illustrate the use of the four novel 8-hydroxydandridine iridium (III) complexes Ro1-Ro4 with high in vitro and in vivo activities in the pharmaceutical industry, the applicant conducted in vitro and in vivo anti-tumor activity experiments on them.

[0049] 1. Experimental study on the proliferation inhibition of three human cell lines by four novel 8-hydroxydandrogenine iridium (III) complexes Ro1-Ro4 with high in vitro and in vivo activity

[0050] 1. Cell lines and cell culture

[0051] In this experiment, three human cell lines were selected, including human ovarian cancer cell line SK-OV-3, cisplatin-resistant line SK-OV-3 / DDP and normal HL-7702 liver cell line.

[0052] All human cell lines were cultured in RPMI-1640 medium containing 100 U / mL penicillin, 10 wt% calf blood, and 100 U / mL streptomycin at 37°C with 5% CO 2 Culture in an incubator.

[0053] 2. Preparation of test compounds

[0054] The purity of all compounds used must be ≥95%. Their DMSO stock solutions are diluted with physiological buffer to a final solution of 20 μmol / L (final DMSO concentration ≤1%), and the degree of inhibition of the growth of normal cells or selected tumor cells by each compound at this concentration is tested.

[0055] 3. Cell Growth Inhibition Experiment

[0056] The specific steps are as follows:

[0057] (1) Cell digestion, counting, and preparation to a concentration of 5×10 5 100 μL of cell suspension was added to each well of a 96-well cell culture plate and placed at 37°C with 5% CO 2 Culture in an incubator for 24 h.

[0058] (2) The drug was diluted to the required concentration using RPMI-1640 medium. 100 μL of the corresponding drug-containing medium was added to each well of the 96-well plate to make the final concentrations reach 0, 1.0, 2.5, 5.0, 10.0, 20.0 and 50.0 μM, respectively. Six parallel experiments were set up for each concentration. After adding the drug, the plate was placed at 37°C and 5% CO 2 The cells were cultured in the incubator for 48 h.

[0059] (3) Perform CCK-8 staining and measure the OD value at a wavelength of λ = 450nm: add 10μL CCK-8 to each well, continue to culture in the incubator for 2h, shake for 10min to gently mix, then measure the OD value of each well at a wavelength of λ = 450nm using an enzyme marker to calculate the inhibition rate of each compound on the growth of the selected cell line, and then calculate the IC value of each test compound on the selected cell line using the Bliss method. 50 The results are shown in Table 1 below.

[0060] Table 1. IC values ​​of compounds against various cell lines 50 Value (μM)

[0061]

[0062] From IC 50 The results of activity screening showed that complex Ro2 had a good anticancer effect on SK-OV-3 and SK-OV-3 / DDP cancer cells, and its IC 50 The values ​​were 6.39±0.80μM and 4.03±0.19μM, especially for SK-OV-3 / DDP cancer cells, and their activity was much greater than that of H-Rob1 (30.27±0.99μM), H-Rob2 (36.11±0.23μM), Ro1 (7.08±0.50μM), Ro3 (13.82±1.04μM), Ro4 (10.16±0.31μM) and cisplatin (>50μM); and Ro1-Ro4 had little inhibitory effect on normal HL-7702 cells (IC 50 value>50μM); indicating that the complexes Ro1-Ro4 can overcome cisplatin resistance and target the inhibition of SK-OV-3 / DDP cancer cell proliferation.

[0063] 2. In vivo tumor inhibition experiment in tumor-bearing nude mice

[0064] Human ovarian cancer cells SK-OV-3 in the logarithmic growth phase were collected and the cells were adjusted to 5×10 8 Use a 1.0mL syringe to extract 0.2mL of the suspension, which contains approximately 1×10 8 The live cells were then inoculated into the subcutaneous tissue of the right axilla of nude mice. The subcutaneous tumors grew to about 1 cm. 3 The human ovarian cancer cells SK-OV-3 were cultured in nude mice for 3-4 generations. After the growth of the cells stabilized, the tumor-bearing mice with vigorous tumor growth and no ulceration were selected and killed by cervical dislocation. The animal skin was disinfected with 75% medical alcohol, the tissue blocks were dissected, the necrotic parts were removed, and the tumor tissue was cut into 1.5mm 3 The left and right small pieces were inoculated into the subcutaneous part of the right armpit of nude mice using a trocar. The diameter of the transplanted tumor was measured with an electronic vernier caliper. When the tumor volume grew to 100-150mm 3 The animals were randomly divided into groups.

[0065] Nude mice bearing human ovarian cancer cell SK-OV-3 tumors were randomly divided into a vehicle group, a drug-added group, and a cisplatin positive control group, with 7 animals in each group. Intraperitoneal injection was started on the day of grouping, with the compound administered every two days and cisplatin administered every other day. The tumor diameter was measured with an electronic vernier caliper every three days, and the body weight was measured. On the 13th day, the mice were killed by cervical dislocation, the tumors were dissected, weighed, photographed, and the tumor inhibition rate was calculated.

[0066] Tumor volume calculation formula: V = a × b 2 / 2, where a is the major diameter and b is the minor diameter;

[0067] Relative tumor volume RTV = V t / V 0 , where V t is the volume at each measurement, V 0 is the volume when grouped;

[0068] Relative tumor proliferation rate T / C% = (T RTV / C RTV )×100%;

[0069] Tumor growth inhibition rate (%) = (average tumor weight of vehicle group - average tumor weight of treatment group) / average tumor weight of vehicle group × 100%.

[0070] Table 2. In vivo tumor inhibition results of complex Ro2 on human ovarian cancer cell SK-OV-3 in nude mice

[0071]

[0072] From the in vivo tumor inhibition experiment shown in Table 2, the in vivo tumor inhibition experiment shows that the complex Ro2 has a good tumor inhibition effect on the nude mouse model of human ovarian cancer cell SK-OV-3, and the inhibition rate is as high as 60.4%. In conclusion, the new 8-hydroxy-dandridine iridium (III) complexes Ro1-Ro4 show excellent in vitro and in vivo antitumor activity, have potential medicinal value, and are expected to be used in the preparation of various antitumor drugs.

[0073] In summary, the 8-hydroxy-dandridine iridium (III) complexes Ro1-Ro4 provided by the present invention exhibit excellent in vitro and in vivo antitumor activity and selectivity, as well as good cytotoxic selectivity, indicating that the design ideas and synthesis methods of the synthesized 8-hydroxy-dandridine iridium (III) complexes of the present invention are feasible. The antitumor activity exhibited by the 8-hydroxy-dandridine iridium (III) complex Ro2 makes it have good potential medicinal value and is expected to be used in the preparation of various antitumor drugs.

Claims

1. 8-Hydroxy-dipsatin iridium (III) complex, characterized in that The chemical structure is shown below:

2. The method for synthesizing the 8-hydroxy-dandrogenine iridium (III) complex according to claim 1, characterized in that: Weigh 2-phenylpyridine iridium (III) dimer or its derivatives and place them in a drug-resistant tube, add 8-hydroxyquinoline or its derivatives, then add a solvent, cover the tube, react at 45°C for 2 days, react at 65°C for 1 day, and finally react at 80°C for 1 day. After the reaction, cool and cut to 25°C to obtain the tube.

3. The synthetic method of 8-hydroxy-dandrogenine iridium (III) complex according to claim 2, characterized in that: The solvent was 2.0 mL MeOH and 0.1 mL DMSO or 2.0 mL MeOH and 0.1 mL CH2Cl2.

4. The method for synthesizing the 8-hydroxydandrogenine iridium (III) complex according to claim 2, characterized in that: The 8-hydroxyquinoline derivative is 8-hydroxybactamine; the 2-phenylpyridine iridium (III) dimer derivative is 3-methyl-2-phenylpyridine iridium (III) dimer.

5. Use of the 8-hydroxydandrogenine iridium (III) complex according to claim 1 in the preparation of drugs for targeted treatment of ovarian cancer.

6. Use of the 1-hydroxy-dandrogenic iridium (III) complex of claim 1 in the preparation of a drug for targeted treatment of drug-resistant ovarian cancer strains.

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