Ru(II) complex with anticancer activity and preparation method of complex

Abstract

The invention provides a Ru(II) complex with anticancer activity and a preparation method of the complex. The complex is [Ru(phen)2GFLX]Cl2. The preparation method of the complex comprises the following steps of: sequentially adding [Ru(phen)2Cl2].2H2O, GFLX and sodium ethoxide under the protection of argon; stirring and dissolving with absolute ethanol, refluxing, adding hydrochloric acid to remove excessive sodium ethoxide, and carrying out spin drying to recycle the solvent; dissolving the obtained solid by using acetonitrile, filtering, carrying out spin-drying on the filtrate to obtain dark brownish red solid; and dissolving the solid by using the acetonitrile, performing neutral alumina column chromatography, eluting with a mixed solution of acetonitrile and ethanol, collecting the brownish red zone, performing spin-drying, and drying in vacuum to obtain the target complex. The Ru(II) complex has a good anticancer effect, and has low injury of normal cells.

Description

technical field

[0001] The invention relates to a metal complex, in particular to a Ru(II) complex with anticancer activity and a preparation method thereof. Background technique

[0002] There are three ways of interaction between metal complexes and DNA, they are 1, electrostatic binding; 2, channel binding; 3, intercalation binding. Electrostatic binding refers to the combination of positively charged metal complexes and negatively charged phosphates in DNA in a manner of electrostatic attraction; channel binding refers to the binding of complexes to the channel site of double-helix DNA; insertional binding refers to the use of complexes The planar aromatic heterocyclic ring it contains is inserted into the base pair of DNA and stacked with its base pair. Among the three methods, the third method is the best combination method, because it can be inserted into the base pair of DNA, directly damaging the base pair, and in genetic diseases such as cancer, due to the mismatc...

Examples

Embodiment 1

[0019] The preparation of embodiment 1Ru (II) complex:

[0020] The first step: the RuCl 3 ·H 2 O1g (4.82mmol), 1,10-phenanthroline (phen) 1.91g (9.64mmol) was dissolved in 30mlN, N'--dimethylformamide, under the protection of argon, heated to reflux for 3h, and then removed by rotary evaporation Most of the solvent, the remaining solution was cooled to room temperature, 250ml of acetone was added, and placed at 0°C overnight. The resulting crystals were filtered off with suction and washed with ice water. Then dissolve the crude product in a mixed solvent of ethanol and water (1:1) and heat to reflux for 1 h, filter the resulting solution to remove insoluble matter, add 23g LiCl to the filtrate, and remove the ethanol in the solution by rotary evaporation, and cool the remaining aqueous solution in an ice bath to obtain ink. Green crystals. Wash with ether and dry in vacuum;

[0021] Step 2: Under argon protection, add 0.0488g (0.2mmol) of the ligand L obtained in the fi...

Embodiment 2

[0026] Example 2 Ru (II) complex anticancer activity experiment

[0027] The anticancer activity was tested by cytotoxicity by using the tetrazolium salt 3-(4,5-dimethylthiazole)-2,5-diphenyltetrazolium bromide (MTT) as the dye. The complex was prepared with phosphoric acid solution and stored in a refrigerator at 4°C for later use. Serial stock solutions were prepared in culture medium in 96-well microtiter plates. The complex was added to the cell culture medium in a concentration gradient (1nM-1μM), and the medium did not need to be replaced within 24 hours. At the end of the set reaction time, the medium was transferred to 200 μL of medium containing 0.5 mg / ml MTT and the microtiter plate was placed in an incubator at 37° C. for 4 hours. 100 μl DMSO was added to the medium to increase the solubility of this purple dye. Finally, the product is obtained by measuring the optical density of each well under 595nm fluorescence. Inhibition rate (II%) was calculated by the fol...

Embodiment 3

[0029] Example 3 Interaction experiment between Ru(II) complexes and DNA

[0030] Absorption Spectroscopy Research

[0031] figure 2 Curve 1 is the ultraviolet absorption spectrum of the Ru(II) complex, and curve 2-17 is the ultraviolet absorption spectrum of the Ru(II) complex after adding DNA. It can be seen from the figure that the absorption peak intensity of the complex decreases when DNA is added. According to literature reports [10] , when the small molecule is intercalated between the base pairs of the CT-DNA double helix, its absorption spectrum shows a color reduction effect; when the small molecule is electrostatically combined with the CT-DNA, its absorption spectrum shows a color enhancement effect ; Therefore, the interaction between Ru(II) complexes and CT-DNA at this time should be dominated by intercalation.

[0032] Effects of Complexes on the Fluorescence Spectrum of DNA

[0033] image 3Curve 1 is the fluorescence spectrum of the Ru(II) complex, and c...

technical field

[0001] The invention relates to a metal complex, in particular to a Ru(II) complex with anticancer activity and a preparation method thereof. Background technique

[0002] There are three ways of interaction between metal complexes and DNA, they are 1, electrostatic binding; 2, channel binding; 3, intercalation binding. Electrostatic binding refers to the combination of positively charged metal complexes and negatively charged phosphates in DNA in a manner of electrostatic attraction; channel binding refers to the binding of complexes to the channel site of double-helix DNA; insertional binding refers to the use of complexes The planar aromatic heterocyclic ring it contains is inserted into the base pair of DNA and stacked with its base pair. Among the three methods, the third method is the best combination method, because it can be inserted into the base pair of DNA, directly damaging the base pair, and in genetic diseases such as cancer, due to the mismatc...

Examples

Embodiment 1

[0019] The preparation of embodiment 1Ru (II) complex:

[0020] The first step: the RuCl 3 ·H 2 O1g (4.82mmol), 1,10-phenanthroline (phen) 1.91g (9.64mmol) was dissolved in 30mlN, N'--dimethylformamide, under the protection of argon, heated to reflux for 3h, and then removed by rotary evaporation Most of the solvent, the remaining solution was cooled to room temperature, 250ml of acetone was added, and placed at 0°C overnight. The resulting crystals were filtered off with suction and washed with ice water. Then dissolve the crude product in a mixed solvent of ethanol and water (1:1) and heat to reflux for 1 h, filter the resulting solution to remove insoluble matter, add 23g LiCl to the filtrate, and remove the ethanol in the solution by rotary evaporation, and cool the remaining aqueous solution in an ice bath to obtain ink. Green crystals. Wash with ether and dry in vacuum;

[0021] Step 2: Under argon protection, add 0.0488g (0.2mmol) of the ligand L obtained in the fi...

Embodiment 2

[0026] Example 2 Ru (II) complex anticancer activity experiment

[0027] The anticancer activity was tested by cytotoxicity by using the tetrazolium salt 3-(4,5-dimethylthiazole)-2,5-diphenyltetrazolium bromide (MTT) as the dye. The complex was prepared with phosphoric acid solution and stored in a refrigerator at 4°C for later use. Serial stock solutions were prepared in culture medium in 96-well microtiter plates. The complex was added to the cell culture medium in a concentration gradient (1nM-1μM), and the medium did not need to be replaced within 24 hours. At the end of the set reaction time, the medium was transferred to 200 μL of medium containing 0.5 mg / ml MTT and the microtiter plate was placed in an incubator at 37° C. for 4 hours. 100 μl DMSO was added to the medium to increase the solubility of this purple dye. Finally, the product is obtained by measuring the optical density of each well under 595nm fluorescence. Inhibition rate (II%) was calculated by the fol...

Embodiment 3

[0029] Example 3 Interaction experiment between Ru(II) complexes and DNA

[0030] Absorption Spectroscopy Research

[0031] figure 2 Curve 1 is the ultraviolet absorption spectrum of the Ru(II) complex, and curve 2-17 is the ultraviolet absorption spectrum of the Ru(II) complex after adding DNA. It can be seen from the figure that the absorption peak intensity of the complex decreases when DNA is added. According to literature reports [10] , when the small molecule is intercalated between the base pairs of the CT-DNA double helix, its absorption spectrum shows a color reduction effect; when the small molecule is electrostatically combined with the CT-DNA, its absorption spectrum shows a color enhancement effect ; Therefore, the interaction between Ru(II) complexes and CT-DNA at this time should be dominated by intercalation.

[0032] Effects of Complexes on the Fluorescence Spectrum of DNA

[0033] image 3Curve 1 is the fluorescence spectrum of the Ru(II) complex, and c...

Images