CN113583028A - High-cancer-cell-selectivity anticancer drug based on metal organic complex and preparation method thereof - Google Patents

Abstract

The invention discloses a high-cancer-cell-selectivity anticancer drug based on a metal organic complex and a preparation method thereof, belonging to the technical field of anticancer drugs. The metal ion and the organic ligand form a coordination bond; the valence of the metal ion is not more than + 2; the coordination monomer of the organic ligand comprises at least one halogen element; at least three coordinating elements A forming coordination bonds with metal ions, said A comprising an element of N and/or an element of O; at least one bonding element B forming hydrogen bonds with other coordinating units, the B comprising an N element and/or an O element; at least one N element which does not form a coordination bond with the metal ion but can form a hydrogen bond with other coordination monomers, the N element being on the heterocycle of the pyrimidine. The complex has strong anticancer performance, has low toxicity to normal cells, and shows excellent cancer cell selectivity.

Description

High-cancer-cell-selectivity anticancer drug based on metal organic complex and preparation method thereof

Technical Field

The invention belongs to the technical field of anti-cancer drugs, and particularly relates to a high-cancer-cell-selectivity anti-cancer drug based on a metal organic complex and a preparation method thereof.

Background

Day 2, 4 of each year is a worldwide cancer day, and establishment of a worldwide cancer day shows a global consensus to overcome the promise and expectation of cancer. Cancer has become the leading cause of death in china, and morbidity and mortality are rising, posing a significant threat to public health. Statistically, over 280 million people died from cancer in 2015, averaging 7500 people per day. The search for effective anti-cancer drugs has been one of the major challenges facing medicinal chemists. Due to the drug resistance and toxicity of the platinum-based complex, in recent years, the copper complex is considered as a potential anti-tumor drug and can well replace the platinum-based complex. Halogen-containing compounds are always paid attention to by medicinal chemists as important anti-cancer drugs, and the research of synthesizing novel organic halogen-containing compounds and finding halogen-containing compounds with good anti-cancer effects is one of the research hotspots of medicinal chemistry.

Through retrieval, Chinese invention patent CN107089996A discloses an anticancer drug 5-bromosalicylaldehyde-2-chloro-6-hydrazinopyridine schiff base copper complex and a synthetic method thereof, and particularly discloses that the chemical formula of the complex is [ Cu (bchp ]₂]·2H₂O·C₂H₅OH, molecular formula C₂₄H₁₈Br₂Cl₂CuN₆O₂·2H₂O·C₂H₅OH with molecular weight of 798.80, wherein Hbchp is 5-bromosalicylaldehyde-2-chloro-6-hydrazino-pyridine Schiff base; said [ Cu (bchp)₂]·2H₂O·C₂H₅The OH was synthesized by dissolving 2.010g of analytically pure 5-bromosalicylaldehyde, 1.436g of analytically pure 2-chloro-6-hydrazinopyridine in 30mL of analytically pure ethanol solution under heating and refluxingThe ligand Hbchp was obtained after two hours. Dissolving dried 0.163-0.326g Hbchp in 5-10mL of analytically pure ethanol, dissolving 0.121-0.242g analytically pure copper nitrate trihydrate in 5-10mL of redistilled water, placing in a reaction kettle, standing in an oven at 80 ℃ for three days, and generating yellow stripe crystals (Cu (bchp)₂]·2H₂O·C₂H₅And (5) OH. Although the anticancer drugs have certain anticancer effect on cancer cells such as BEL-7404 human liver cancer cell line, Hep-G2 cancer cell line and the like, the anticancer drugs still have non-negligible toxicity on normal cells.

Therefore, there is a need to design an anticancer drug with high selectivity to cancer cells and low toxicity to normal cells or a preparation method thereof.

Disclosure of Invention

1. Problems to be solved

Aiming at the problems that the anticancer drug in the prior art has lower anticancer performance and nonnegligible toxicity to normal cells, the invention provides a high-cancer-cell-selectivity anticancer drug based on a metal organic complex and a preparation method thereof; by reasonably designing the structure of the metal organic complex, the metal organic complex can effectively solve the problems of low cancer performance and inconspicuous toxicity to normal cells when being used as an anti-cancer medicament.

2. Technical scheme

In order to solve the problems, the technical scheme adopted by the invention is as follows:

the invention relates to a high-cancer-cell-selectivity anticancer drug based on a metal organic complex, which comprises metal ions and organic ligands, wherein the metal ions and the organic ligands form coordinate bonds; the valence of the metal ion is not more than + 2;

the coordination monomer of the organic ligand comprises

At least one halogen element;

at least three coordinating elements A forming coordination bonds with metal ions, said A comprising an element of N and/or an element of O;

at least one bonding element B forming hydrogen bonds with other coordinating units, the B comprising an N element and/or an O element;

at least one N element which does not form a coordination bond with the metal ion but can form a hydrogen bond with other coordination monomers, the N element being on the heterocycle of the pyrimidine.

Preferably, the metal ion complex also comprises an oxygen-containing solvent ligand, wherein a coordination monomer X of the oxygen-containing solvent ligand forms a coordination bond with a metal ion through an oxygen element; the coordination monomer X of the oxygenated solvent ligand comprises H₂O and/or MeOH.

Preferably, the metal ions comprise Cu²⁺And/or Cu⁺。

Preferably, the metal ion is Cu²⁺The anticancer drug monomer has the chemical formula 1:

in chemical formula 1, X₁、X₂Are the same or different and are each independently H₂O or MeOH, X₃Is H₂O or MeOH; r¹～R⁷The same or different and are each independently H or C1-C10 alkyl or carboxyl or amino or hydroxyl or halogen or methoxy or ethoxy or nitro.

Preferably, the coordination monomers of the organic ligand comprise coordination monomer M and coordination monomer N; the coordination monomer M, the coordination monomer N and the coordination monomer X containing oxygen ligand are respectively as follows:

and H₂O; or

And H₂O, MeOH, respectively; or

And H₂O; or

And H₂O; or

And MeOH; or

And H₂O; or

And H₂O。

Preferably, the molar ratio of the coordination monomer M to the coordination monomer N is (0.7-1.4): 1.

the invention relates to a preparation method of a high-cancer-cell-selectivity anticancer drug based on a metal organic complex, wherein the anticancer drug prepared by the method is the anticancer drug in the invention, and a compound of a chemical formula 2 and a compound of a chemical formula 3 are reacted to obtain an organic ligand of the anticancer drug;

the chemical formula 2 is:

the chemical formula 3 is:

in chemical formula 3, R¹Is H or Br or Cl or OCH₃Or OCH₂CH₃，R³Is H or Br or Cl.

8. The preparation method of the metal organic complex-based high cancer cell selectivity anticancer drug according to claim 7, characterized by comprising the following specific steps:

(1) mixing a compound of a chemical formula 2 and a compound of a chemical formula 3, dissolving the mixture in methanol, and heating and refluxing the mixture to obtain a coordination monomer M;

(2) dissolving the dried coordination monomer M in a solvent, adding a copper ion salt, and stirring for dissolving to obtain a mixed solution;

(3) placing the mixed solution obtained in the step (2) into a reaction kettle, and then placing the reaction kettle into an oven for standing reaction to obtain the anti-cancer drug;

in the above steps (1) to (3), the structural formula of the coordination monomer M is:

9. the method of claim 8, wherein the concentration of the compound of formula 2 in step (1) is 0.2-0.8 mol/L, the concentration of the compound of formula 3 in step (1) is 0.24-0.96 mol/L, the temperature of the heating reflux is 70-90 ℃ and the time is 3-5 hours.

10. The method of claim 8, wherein the solvent used in step (2) comprises 1-4 mL of analytically pure methanol, 0-3 mL of analytically pure DMF, and 3-9 mL of deionized water; the copper ion salt is copper nitrate trihydrate; the stirring time is 0.5 to 3 hours; the concentration of coordination monomer M in the mixed solution is 12.0 mg/mL-27.3 mg/mL, and the concentration of copper ions is 2.9 mg/mL-7.1 mg/mL;

and/or the standing reaction temperature in the step (3) is 65-90 ℃, and the reaction time is 48-96 hours.

3. Advantageous effects

Compared with the prior art, the invention has the beneficial effects that:

(1) the invention relates to a high-cancer-cell-selectivity anticancer drug based on a metal organic complex, which comprises metal ions and organic ligands, wherein the metal ions and the organic ligands form coordinate bonds; the valence of the metal ion is not more than + 2; the coordination monomer of the organic ligand comprises at least one halogen element; at least three coordinating elements A forming coordination bonds with metal ions, said A comprising an element of N and/or an element of O; at least one bonding element B forming hydrogen bonds with other coordinating units, the B comprising an N element and/or an O element; at least one N element which does not form a coordination bond with a metal ion, but can form a hydrogen bond with other coordination monomers, the N element being on the heterocycle of the pyrimidine; through the arrangement, the complex with metal ions and organic ligands continuously coordinated can be obtained, and the complex has strong anticancer performance, has low toxicity to normal cells and shows excellent cancer cell selectivity.

(2) The anti-cancer drugs provided by the invention have excellent anti-cancer performance and cancer cell selectivity, wherein the inhibition rate of the complex 4 on BEL-7404 human hepatoma cell line, Hep-G2 cancer cell line and Hela cervical carcinoma cell can reach 70.82 +/-0.39, 79.06 +/-0.36 and 82.69 +/-0.39; it has IC effect on BEL-7404 human liver cancer cell line, Hep-G2 cancer cell line and Hela cervical cancer cell₅₀The values are respectively 10.32 + -0.35 μ M, 5.01 + -0.33 μ M and 2.05 + -0.33 μ M, and the IC of the published invention patent CN107089996A for BEL-7404 human liver cancer cell line and Hep-G2 cancer cell line₅₀The values are respectively 10.34 +/-1.06 mu M and 8.56 +/-1.50 mu M, and the activity of the complex 4 to BEL-7404 human liver cancer cell line and Hep-G2 cancer cell line is obviously higher than that of the prior art; meanwhile, the complex of the invention has IC of 4 pairs of HL-7702 normal liver cells₅₀Maintaining at above 100 μ M, and IC of published invention patent CN107089996A on HL-7702 normal liver cell₅₀Reaches 36.09 +/-0.71 mu M, so the anticancer drug of the invention shows excellent cancer cell selectivity.

Drawings

FIG. 1 is a schematic diagram of the synthesis of anticancer drugs according to embodiments 1 to 7 of the present invention;

FIG. 2 is a schematic structural diagram of complex 1 prepared in example 1 of the present invention;

FIG. 3 is a three-dimensional stacking diagram of complex 1 prepared in example 1 of the present invention;

FIG. 4 is a schematic structural diagram of complex 2 prepared in example 2 of the present invention;

FIG. 5 is a three-dimensional stacking diagram of complex 2 prepared in example 2 of the present invention;

FIG. 6 is a schematic structural diagram of complex 3 prepared in example 3 of the present invention;

FIG. 7 is a three-dimensional stacking diagram of complex 3 prepared in example 3 of the present invention;

FIG. 8 is a schematic structural diagram of complex 4 prepared in example 4 of the present invention;

FIG. 9 is a three-dimensional stacking diagram of complex 4 prepared in example 4 of the present invention;

FIG. 10 is a schematic structural diagram of complex 5 prepared in example 5 of the present invention;

FIG. 11 is a three-dimensional stacking diagram of complex 5 prepared in example 5 of the present invention;

FIG. 12 is a schematic structural diagram of complex 6 prepared in example 6 of the present invention;

FIG. 13 is a three-dimensional stacking diagram of complex 6 prepared in example 6 of the present invention;

FIG. 14 is a schematic structural diagram of complex 7 prepared in example 7 of the present invention;

FIG. 15 is a three-dimensional stacking diagram of complex 7 prepared in example 7 of the present invention.

Detailed Description

The following detailed description of exemplary embodiments of the invention refers to the accompanying drawings, which form a part hereof, and in which are shown by way of illustration exemplary embodiments in which the invention may be practiced, and in which features of the invention are identified by reference numerals. The following more detailed description of the embodiments of the invention is not intended to limit the scope of the invention, as claimed, but is presented for purposes of illustration only and not limitation to describe the features and characteristics of the invention, to set forth the best mode of carrying out the invention, and to sufficiently enable one skilled in the art to practice the invention. It will, however, be understood that various modifications and changes may be made without departing from the scope of the invention as defined in the appended claims. The detailed description and drawings are to be regarded as illustrative rather than restrictive, and any such modifications and variations are intended to be included within the scope of the present invention as described herein. Furthermore, the background is intended to be illustrative of the state of the art as developed and the meaning of the present technology and is not intended to limit the scope of the invention or the application and field of application of the invention.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs; the terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention; as used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

The invention provides 7 metal organic complexes as anticancer drugs, and the structure, preparation method and anticancer performance of the metal organic complexes are respectively illustrated by each example. As can be seen from FIG. 1, the structural formula and the synthetic raw materials of the anticancer drug in 7 are basically the same, and the main difference is that R is¹、R²And R⁷In contrast, the functional groups are selected in different embodiments as follows:

example 1: r in the reactants¹＝H，R³＝Br，R⁷Cl, R in the product¹＝H，R³＝Br，R⁷＝H；

Example 2: r in the reactants¹＝R³＝Br，R⁷Cl, R in the product¹＝R³＝Br，R⁷＝H；

Example 3: r in the reactants¹＝OCH₃，R³＝H，R⁷Cl, R in the product¹＝OCH₃，R³＝H，R⁷＝H；

Example 4: r in the reactants¹＝OCH₂CH₃，R³＝H，R⁷Cl, R in the product¹＝OCH₂CH₃，R³＝H，R⁷＝H；

Example 5: r in the reactants¹＝R³＝Cl，R⁷Cl, R in the product¹＝R³＝Cl，R⁷＝H；

Example 6: r in the reactants¹＝R³＝H，R⁷Cl, R in the product¹＝R³＝H，R⁷＝H；

Example 7: r in the reactants¹＝NO₂，R³＝H，R⁷Cl, R in the product¹＝NO₂，R³＝H，R⁷＝H。

The crystallographic data of the complexes in the examples of the invention are shown in tables 1 and 2, wherein complex 1 was prepared by the preparation method of example 1, and the rest is similar.

TABLE 1, data sheet of crystallography of complexes 1-3

^aR₁＝Σ||F_o|–|F_c||/Σ|F_o|.^b wR₂＝[Σw(|F_o ²|–|F_c ²|)²/Σw(|F_o ²|)²]^1/2

TABLE 2 crystallographic data table of complexes 4-7

^aR₁＝Σ||F_o|–|F_c||/Σ|F_o|.^bwR₂＝[Σw(|F_o ²|–|F_c ²|)²/Σw(|F_o ²|)²]^1/2

The invention is further described with reference to specific examples.

Example 1

This example provides a high cancer cell selectivity anticancer drug based on metal organic complex, in this example, complex 1, whose schematic structural diagram and three-dimensional stacking diagram are shown in fig. 2 and fig. 3. The chemical formula of the complex 1 is expressed as

[Cu(Hbhp)_0.49(bchp)_0.51(H₂O)]Wherein M is 5-bromosalicylaldehyde-2-chloro-4-hydrazinopyrimidine Schiff base in this example, represented by Hbchp, bchp is dehydrogenated Hbchp, and N is 5-bromosalicylaldehyde-4-hydrazinopyrimidine Schiff base in this example (generated in situ by the reaction system of Hbchp and copper ions), represented by Hbhp. The formula of the complex 1 is thus C₁₁H_8.49BrCl_0.51CuN₄O₂The molecular weight is 390.24, the crystal structure data is shown in Table 1, and the bond length and bond angle data is shown in Table 3.

TABLE 3 partial bond length of Complex 1

Angle of harmony key (°)

As shown in FIG. 1, the present embodiment further provides a method for preparing a metal-organic complex-based anticancer drug with high cancer cell selectivity [ Cu (Hbhp) ]_0.49(bchp)_0.51(H₂O)]The synthesis method comprises the following specific steps:

12mmol (2.412g) of analytically pure 5-bromosalicylaldehyde and 10mmol of 1.4456g of analytically pure 2-chloro-4-hydrazinopyrimidine are dissolved in 50mL of analytically pure methanol solution and heated under reflux for 4 hours to give the ligand Hbchp. Dissolving dried 0.146-0.292g Hbchp in 3mL of analytically pure methanol, 3mL of analytically pure DMF and 3mL of deionized water, adding 0.121-0.242g analytically pure copper nitrate trihydrate, stirring for 1 hour, placing in a reaction kettle, standing in an oven at 80 ℃ for three days to obtain yellow blocky crystals, namely [ Cu (Hbhp ]_0.49(bchp)_0.51(H₂O)]。

Example 2

This example provides a high cancer cell selectivity anticancer drug based on metal organic complex, in this example, complex 2, which is a schematic structural diagramAnd three-dimensional stacking diagrams as shown in fig. 4 and 5. The chemical formula of the complex 2 is expressed as [ Cu (Hdhp) ]_0.5(Hdbchp)_0.5(H₂O)(MeOH)]·(NO₃) Wherein M is 3, 5-dibromo salicylaldehyde-condensed-2-chloro-4-hydrazinopyrimidine Schiff base in the embodiment and is represented by Hdbchp, and N is 3, 5-dibromo salicylaldehyde-condensed 4-hydrazinopyrimidine Schiff base in the embodiment (generated in situ by a reaction system of Hdbchp and copper ions) and is represented by Hdhp. The formula of the complex 2 is thus C₁₂H_13.5Br₂Cl_0.5CuN₅O₆Molecular weight is 564.99, and its crystal structure data are shown in Table 1 and bond length and bond angle data are shown in Table 4.

TABLE 4 partial bond length of Complex 2

Angle of harmony key (°)

As shown in FIG. 1, this example also provides a method for preparing a highly cancer cell selective anticancer drug based on a metal organic complex, [ Cu (Hdhp) ]_0.5(Hdbchp)_0.5(H₂O)(MeOH)]·(NO₃) The synthesis method comprises the following specific steps:

12mmol (3.359g) of analytically pure 3, 5-dibromo salicylaldehyde and 10mmol of 1.4456g of analytically pure 2-chloro-4-hydrazinopyrimidine are dissolved in 50mL of analytically pure methanol solution and heated under reflux for 4 hours to obtain the ligand Hdbchp. Dissolving dried 0.204-0.408g Hdbchp in 3mL of analytically pure methanol, 3mL of analytically pure DMF and 3mL of deionized water, adding 0.121-0.242g of analytically pure copper nitrate trihydrate, stirring for 1 hour, placing in a reaction kettle, standing in an oven at 80 ℃ for three days to generate green strip crystals, namely [ Cu (Hdhp) ]_0.5(Hdbchp)_0.5(H₂O)(MeOH)]·(NO₃)。

Example 3

This example provides a highly cancer cell selective anticancer drug based on a metal organic complex, in this example complex 3,the schematic structure and three-dimensional stacking diagram are shown in fig. 6 and 7. The chemical formula of the complex 3 is expressed as [ Cu (mhp) ]_0.46(mchp)_0.54(H₂O)]·(H₂O), wherein M is 3-methoxysalicylaldehyde-condensed-2-chloro-4-hydrazinopyrimidine Schiff base in the present example, represented by Hmchp, the mchp is dehydrogenated Hmchp, and N is 3-methoxysalicylaldehyde-condensed 4-hydrazinopyrimidine Schiff base in the present example (generated in situ by reaction of Hmchp and copper ions), represented by Hmhp, the mhp is dehydrogenated Hmhp. The formula of the complex 3 is thus C₁₂H_14.46Cl_0.54CuN₄O₄Molecular weight is 361.43, and its crystal structure data are shown in Table 1 and bond length and bond angle data are shown in Table 5.

TABLE 5 partial bond length of Complex 3

Angle of harmony key (°)

As shown in FIG. 1, this example also provides a method for preparing a metal organic complex-based anticancer drug with high cancer cell selectivity [ Cu (mhp) ]_0.46(mchp)_0.54(H₂O)]·(H₂O) comprises the following steps:

12mmol (1.8259g) of analytically pure 3-methoxysalicylaldehyde and 10mmol of 1.4456g of analytically pure 2-chloro-4-hydrazinopyrimidine are dissolved in 50mL of analytically pure methanol solution and heated under reflux for 4 hours to give the ligand Hmchp. Dissolving dried 0.132-0.264g Hmchp in 4mL of analytically pure methanol, 3mL of analytically pure DMF and 4mL of deionized water, adding 0.121-0.242g of analytically pure copper nitrate trihydrate, stirring for 1 hour, placing in a reaction kettle, standing in an oven at 80 ℃ for three days to obtain black blocky crystals, namely [ Cu (mhp) ]_0.46(mchp)_0.54(H₂O)]·(H₂O)。

Example 4

This example provides a metal organic complex-based highly cancer cell selective anticancer agentThe complex 4 in this example has a structure diagram and a three-dimensional stacking diagram as shown in FIGS. 8 and 9. The chemical formula of the complex 4 is expressed as [ Cu (ehp) ]_0.49(echp)_0.51(H₂O)]·(H₂O), wherein M is 3-ethoxysalicylaldehyde-2-chloro-4-hydrazinopyrimidine Schiff base in the present example, represented by Hechp, echp is dehydrogenized Hechp, N is 3-ethoxysalicylaldehyde-4-hydrazinopyrimidine Schiff base in the present example (produced in situ by the reaction system of Hechp and copper ions), represented by Hehp, and ehp is dehydrogenized Hehp. The formula of complex 4 is thus C₁₃H_16.49Cl_0.51CuN₄O₄The molecular weight is 374.42, the crystal structure data is shown in Table 2, and the bond length and bond angle data is shown in Table 6.

TABLE 6 partial bond length of Complex 4

Angle of harmony key (°)

As shown in FIG. 1, this example also provides a method for preparing a metal organic complex-based anticancer drug with high cancer cell selectivity, which is [ Cu (ehp) ]_0.49(echp)_0.51(H₂O)]·(H₂O) comprises the following steps:

12mmol (1.994g) of analytically pure 3-ethoxysalicylaldehyde and 10mmol of 1.4456g of analytically pure 2-chloro-4-hydrazinopyrimidine are dissolved in 50mL of analytically pure methanol solution and heated under reflux for 4 hours to give the ligand Hechp. Dissolving dried 0.146-0.292g Hechp in 1mL of analytically pure methanol and 9mL of deionized water, adding 0.121-0.242g of analytically pure copper nitrate trihydrate, stirring for 1 hour, placing in a reaction kettle, standing in an oven at 80 ℃ for three days, and generating black blocky crystals (Cu (ehp))_0.49(echp)_0.51(H₂O)]·(H₂O)。

Example 5

The present embodiment provides a metal-organic based formulationThe high cancer cell selective anticancer drug, in this example, is complex 5, and the schematic structural diagram and the three-dimensional stacking diagram thereof are shown in fig. 10 and fig. 11. The chemical formula of the complex 5 is expressed as [ Cu (bchp)_0.42(bcchp)_0.58(MeOH)]Wherein M is 3, 5-dichlorosalicylaldehyde-condensed-2-chloro-4-hydrazinopyrimidine Schiff base expressed by Hbcchp in the embodiment, bcchp is dehydrogenated Hbcchp, N is 3, 5-dichlorosalicylaldehyde-condensed 4-hydrazinopyrimidine Schiff base expressed by Hbchp in the embodiment (Hbcchp and copper ion reaction system generated in situ), and bchp is dehydrogenated Hbchp. The formula of complex 5 is thus C₁₂H_10.42Cl_2.58CuN₄O₂The molecular weight is 397.67, the crystal structure data is shown in Table 2, and the bond length and bond angle data is shown in Table 7.

TABLE 7 partial bond length of Complex 5

Angle of harmony key (°)

As shown in FIG. 1, this example also provides a method for preparing a highly cancer cell selective anticancer drug based on a metal organic complex, [ Cu (bchp) ]_0.42(bcchp)_0.58(MeOH)]The synthesis method comprises the following specific steps:

12mmol (2.292g) of analytically pure 3, 5-dichlorosalicylaldehyde salicylaldehyde and 10mmol of 1.4456g of analytically pure 2-chloro-4-hydrazinopyrimidine are dissolved in 50mL of analytically pure methanol solution and heated under reflux for 4 hours to obtain the ligand Hdcchp. Dissolving dried 0.164-0.328g Hbcchp in 3mL of analytically pure methanol, 3mL of analytically pure DMF and 3mL of deionized water, adding 0.121-0.242g analytically pure copper nitrate trihydrate, stirring for 1 hour, placing in a reaction kettle, standing in an oven at 80 ℃ for three days to generate green needle-like crystals (Cu (bchp)_0.42(bcchp)_0.58(MeOH)]。

Example 6

This example provides a metal-based organic complexThe high cancer cell selective anticancer drug of (2), in this example, is a complex 6, and the schematic structural diagram and the three-dimensional stacking diagram thereof are shown in fig. 12 and 13. The chemical formula of the complex 6 is expressed as [ Cu (hp)_0.52(chp)_0.48(H₂O)]·(H₂O), wherein M is salicylaldehyde-condensed-2-chloro-4-hydrazinopyrimidine Schiff base in the present example, represented by Hchp, chp is dehydrogenated Hchp, N is salicylaldehyde-condensed 4-hydrazinopyrimidine Schiff base in the present example (generated in situ by the reaction system of Hchp and copper ions), represented by Hhp, and hp is dehydrogenated Hhp. The complex 6 is therefore of formula C₁₁H_12.52Cl_0.48CuN₄O₃The molecular weight is 329.34, the crystal structure data is shown in Table 2, and the bond length and bond angle data is shown in Table 8.

TABLE 8 partial bond length of Complex 6

Angle of harmony key (°)

As shown in FIG. 1, this example also provides a method for preparing a metal organic complex-based anticancer drug with high cancer cell selectivity, which is [ Cu (hp) ]_0.52(chp)_0.48(H₂O)]·(H₂O) comprises the following steps:

12mmol (1.2mL) of analytically pure salicylaldehyde, 10mmol of 1.4456g of analytically pure 2-chloro-4-hydrazinopyrimidine dissolved in 50mL of analytically pure methanol solution are heated under reflux for 8 hours to give the ligand Hchp. Dissolving dried 0.115-0.230g Hchp in 4mL of analytically pure methanol, 3mL of analytically pure DMF and 4mL of deionized water, adding 0.121-0.242g of analytically pure copper nitrate trihydrate, stirring for 1 hour, placing in a reaction kettle, standing in an oven at 80 ℃ for three days to obtain green needle crystals, namely [ Cu (hp)_0.52(chp)_0.48(H₂O)]·(H₂O)。

Example 7

The embodiment provides a machine based on metal organic machineryThe high cancer cell selectivity anticancer drug of the compound, which is the complex 7 in this example, is shown in the schematic structure diagram and the three-dimensional stacking diagram in fig. 14 and 15. The chemical formula of complex 7 is { [ Cu (Hnhp) ]_0.5(Hnchp)_0.5(H₂O)₂]·(H₂O)₂·(NO₃)}₂Wherein M is 5-nitrosalicylaldehyde-2-chloro-4-hydrazinopyrimidine Schiff base in this example and is represented by Hnchp, and N is 5-nitrosalicylaldehyde-4-hydrazinopyrimidine Schiff base in this example (the reaction system of Hnchp and copper ions is generated in situ) and is represented by Hnhhp. The complex 7 is therefore of formula C₂₂H₃₃ClCu₂N₁₂O₂₀The molecular weight is 948.15, the crystal structure data is shown in Table 2, and the bond length and bond angle data is shown in Table 9.

TABLE 9 partial bond length of Complex 7

Angle of harmony key (°)

As shown in FIG. 1, this example also provides a method for preparing a metal organic complex-based anticancer drug with high cancer cell selectivity, which is { [ Cu (Hnhp) ]_0.5(Hnchp)_0.5(H₂O)₂]·(H₂O)₂·(NO₃)}₂The synthesis method comprises the following specific steps:

12mmol (2.004g) of analytically pure 5-nitro salicylaldehyde and 10mmol of 1.4456g of analytically pure 2-chloro-4-hydrazinopyrimidine are dissolved in 50mL of analytically pure methanol solution and heated under reflux for 4 hours to give the ligand Hnchp. Dissolving dried 0.131-0.262g Hnchp in 3mL of analytically pure methanol, 3mL of analytically pure DMF and 3mL of deionized water, adding 0.121-0.242g of analytically pure copper nitrate trihydrate, stirring for 1 hour, placing in a reaction kettle, standing in an oven at 80 ℃ for three days to obtain green stripe crystals, namely { [ Cu (Hnhhp)_0.5(Hnchp)_0.5(H₂O)₂]·(H₂O)₂·(NO₃)}₂。

In order to test the anticancer performance of the complexes in examples 1 to 7, the growth inhibition rates (Table 10) and IC (Integrated Circuit) of the complexes 1 to 7 on different cell lines of BEL-7404 human hepatoma cell line, Hep-G2 cancer cell line, Hela cervical cancer cell and HL-7702 normal hepatoma cell line were measured by an MTT method₅₀Values (table 11).

Table 10, complexes 1-7, inhibition rate of copper chloride and cisplatin on human cancer cells under 20 μ M condition

TABLE 11 IC of human cancer cells for Complex 1-7, cupric chloride, cisplatin and drug of comparative example 3₅₀Value (μ M)

Comparative example 1

The comparative example provides cisplatin as a reference substance for anticancer performance test, and the growth inhibition rates (Table 10) and IC of cisplatin on different cell lines such as BEL-7404 human hepatoma cell line, Hep-G2 cancer cell line, Hela cervical cancer cell line, HL-7702 normal hepatoma cell line and the like were measured by MTT method₅₀Values (table 11).

Comparative example 2

This comparative example provides CuCl₂·4H₂O as an anticancer drug, growth inhibition rates (Table 10) and IC of copper chloride on different cell lines such as BEL-7404 human hepatoma cell line, Hep-G2 cancer cell line, Hela cervical cancer cell, HL-7702 normal hepatoma cell line, etc. were measured by MTT method₅₀Values (table 11).

Comparative example 3

The comparative example provides a 5-bromosalicylaldehyde-2-chloro-6 hydrazinopyridine schiff base copper complex and a synthesis method thereof, and the chemical formula of the complex is [ Cu (bchp)₂]·2H₂O·C₂H₅OH, molecular formula C₂₄H₁₈Br₂Cl₂CuN₆O₂·2H₂O·C₂H₅OH, molecular weight: 798.80, wherein Hbchp is 5-bromosalicylaldehyde-2-chloro-6-hydrazino-pyridine Schiff base.

Said [ Cu (bchp)₂]·2H₂O·C₂H₅The OH synthesis method comprises the following specific steps:

2.010g of analytically pure 5-bromosalicylaldehyde, 1.436g of analytically pure 2-chloro-6-hydrazinopyridine were dissolved in 30mL of analytically pure ethanol solution, and heated under reflux for two hours to give the ligand Hbchp. Dissolving dried 0.163-0.326g Hbchp in 5-10mL of analytically pure ethanol, dissolving 0.121-0.242g analytically pure copper nitrate trihydrate in 5-10mL of redistilled water, placing in a reaction kettle, standing in an oven at 80 ℃ for three days, and generating yellow stripe crystals (Cu (bchp)₂]·2H₂O·C₂H₅OH。

The IC of the complex on different cell lines such as BEL-7404 human liver cancer cell line, Hep-G2 cancer cell line, Hela cervical cancer cell, HL-7702 normal liver cell line, etc. is determined by MTT method₅₀Values (table 11).

The specific test method of the invention is as follows:

selected cell lines were in 5% CO₂Culturing in 37 deg.C culture box, wherein the culture solution is DMEM or RPMI-1640 culture solution containing 10% newborn bovine serum and streptomycin 100U/mL each. The growth of the cells was observed using an inverted microscope, and cells were subjected to digestion passage with 0.25% trypsin at the time of inoculation and selected for cytotoxicity test in the logarithmic growth phase. Test tumor cells were inoculated in culture medium containing 10% neonatal calf serum PPMI1640, 5% CO at 37 deg.C₂Culturing in an incubator under saturated humidity conditions, and changing the culture medium for 2 to 3 times per week and subculturing once in 6 to 7 days. The cancer cells in the culture solution were seeded at a concentration of 2X 104 cells/mL in a 96-well cell culture plate at 190. mu.L per well volume, placed at 37 ℃ and 5% CO₂Cultured under the conditions of (1) for one day. Adding 10 μ L of test compound with different concentrations into each well after the cells adhere to the wall, wherein the DMSO concentration is less than or equal to 1%, and setting corresponding negative control group as culture solution without drug, only test cells and DM with same amountAnd (3) SO. For in-well testing, six replicates per sample were used. After two days of incubation, MTT reagent (5mg/mL, 10. mu.L) was added to each well and incubation continued for 4h, after completion of the incubation, the supernatant was aspirated.

On one hand, after 150. mu.L of DMSO was added to each well, the OD value of each well at a wavelength of 490nm was measured using a microplate reader, and the cell proliferation inhibition rate was calculated by the following formula: inhibition (%) ((OD value of control-OD value of sample group)/OD value of control) x 100%; on the other hand, each IC was calculated by the Bliss method₅₀Values, averaged after 3 replicates of all experiments.

After the treatment for 48 hours with the drug concentration of 20 mu M, as can be seen from Table 10, the complexes 1-7 have excellent anticancer performance and low toxicity to normal cells; copper chloride does not show selectivity to cancer cells or any anticancer performance, so the complexes 1-7 in the invention are compared with cisplatin.

Wherein the inhibition rates of 70.82 +/-0.39, 79.06 +/-0.36 and 82.69 +/-0.39 of the complex 4 to BEL-7404, Hep-G2 and Hela are all higher than the inhibition rate of cisplatin of the comparative example 1, the inhibition rate of the complex 3 to HeLa has good selectivity, and the inhibition rate of the complex 5 to HeLa is close to the inhibition rate of cisplatin; slave IC₅₀The test results are consistent with the inhibition rate, and the IC of BEL-7404, Hep-G2 and Hela of the 4-pair complex is determined₅₀The values are respectively 10.32 +/-0.35 mu M, 5.01 +/-0.33 mu M and 2.05 +/-0.33 mu M, which are all lower than the IC corresponding to the cis-platinum₅₀This indicates that only low concentrations of complex 4 are required to achieve an anti-cancer effect compared to cisplatin. In addition, IC of Compound 6 on Hep-G2 and Hela₅₀The values are respectively 10.03 +/-0.36 mu M and 9.02 +/-1.09 mu M, and are also lower than the IC corresponding to the cis-platinum₅₀The compound has better anticancer performance. The experimental result shows that the complex 4 has stronger apoptosis induction capability on BEL-7404 human hepatoma cell lines, Hep-G2 and Hela cervical carcinoma cells, and has lower toxicity on normal cells; on the contrary, cisplatin has certain inhibition on BEL-7404 human liver cancer cell lines, Hep-G2 cancer cell lines, Hela cervical cancer cells and HL-7702 normal liver cell lines, and the toxicity on normal cells is far higher than that of the complexes 1-7. Thus, gold of the present inventionBelongs to an organic complex, ensures stronger anticancer performance, has lower toxicity to normal cells, and is a promising anticancer drug.

In addition, comparative example 3 is carried out according to the preparation method of the anticancer drug in the prior patent, and comparison of comparative example 3 with examples 1 to 7 shows that the anticancer drug prepared in comparative example 3 has certain anticancer performance on BEL-7404 human hepatoma cell lines and Hep-G2 cancer cell lines, but has strong toxicity on HL-7702 normal hepatoma cell lines, which is the fatal defect, cannot show obvious selectivity on cancer cells, and is very likely to cause great damage to human bodies.

More specifically, although exemplary embodiments of the invention have been described herein, the invention is not limited to these embodiments, but includes any and all embodiments modified, omitted, combined, e.g., between various embodiments, adapted and/or substituted, as would be recognized by those skilled in the art from the foregoing detailed description. The limitations in the claims are to be interpreted broadly based the language employed in the claims and not limited to examples described in the foregoing detailed description or during the prosecution of the application, which examples are to be construed as non-exclusive. Any steps recited in any method or process claims may be executed in any order and are not limited to the order presented in the claims. The scope of the invention should, therefore, be determined only by the appended claims and their legal equivalents, rather than by the descriptions and examples given above.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, including definitions, will control. When "mass, concentration, temperature, time, or other value or parameter is expressed as a range, preferred range, or as a range defined by a list of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, a range of 1 to 50 should be understood to include any number, combination of numbers, or subrange selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50, and all fractional values between the above integers, e.g., 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, and 1.9. With respect to sub-ranges, specifically consider "nested sub-ranges" that extend from any endpoint within the range. For example, nested sub-ranges of exemplary ranges 1-50 may include 1-10, 1-20, 1-30, and 1-40 in one direction, or 50-40, 50-30, 50-20, and 50-10 in another direction. "

Claims (10)

1. A high cancer cell selectivity anticancer drug based on a metal organic complex, which is characterized by comprising a metal ion and an organic ligand, wherein the metal ion and the organic ligand form a coordination bond; the valence of the metal ion is not more than + 2;

the coordination monomer of the organic ligand comprises

At least one halogen element;

at least three coordinating elements A forming coordination bonds with metal ions, said A comprising an element of N and/or an element of O;

at least one bonding element B forming hydrogen bonds with other coordinating units, the B comprising an N element and/or an O element;

at least one N element which does not form a coordination bond with the metal ion but can form a hydrogen bond with other coordination monomers, the N element being on the heterocycle of the pyrimidine.

2. The metal-organic complex-based high cancer cell-selective anticancer drug according to claim 1, further comprising an oxygen-containing solvent ligand, wherein the coordination monomer X of the oxygen-containing solvent ligand forms a coordination bond with a metal ion through an oxygen element; the coordination monomer X of the oxygenated solvent ligand comprises H₂O and/or MeOH.

3. The metal-organic complex-based high cancer cell-selective anticancer agent as claimed in claim 2, wherein the metal ion comprises Cu²⁺And/or Cu⁺。

4. The metal-organic complex-based high cancer cell-selective anticancer drug according to claim 3, wherein the metal ion is Cu²⁺The anticancer drug monomer has the chemical formula 1:

in chemical formula 1, X₁、X₂Are the same or different and are each independently H₂O or MeOH, X₃Is H₂O or MeOH; r¹～R⁷The same or different and are each independently H or C1-C10 alkyl or carboxyl or amino or hydroxyl or halogen or methoxy or ethoxy or nitro.

5. The metal-organic complex-based high cancer cell-selective anticancer drug according to claim 4, wherein the coordination monomer of the organic ligand comprises coordination monomer M and coordination monomer N; the coordination monomer M, the coordination monomer N and the coordination monomer X containing oxygen ligand are respectively as follows:

and H₂O; or

And H₂O, MeOH, respectively; or

And H₂O; or

And H₂O; or

And MeOH; or

And H₂O; or

And H₂O。

6. The metal-organic complex-based high-cancer-cell-selectivity anticancer drug as claimed in claim 5, wherein the molar ratio of coordination monomer M to coordination monomer N is (0.7-1.4): 1.

7. a method for preparing an anticancer drug with high cancer cell selectivity based on a metal organic complex, the anticancer drug prepared by the method is the anticancer drug according to any one of claims 1 to 6, and the method is characterized in that a compound of a chemical formula 2 and a compound of a chemical formula 3 are reacted to obtain an organic ligand of the anticancer drug;

the chemical formula 2 is:

the chemical formula 3 is:

in chemical formula 3, R¹Is H or Br or Cl or OCH₃Or OCH₂CH₃，R³Is H or Br or Cl.

8. The preparation method of the metal organic complex-based high cancer cell selectivity anticancer drug according to claim 7, characterized by comprising the following specific steps:

(1) mixing a compound of a chemical formula 2 and a compound of a chemical formula 3, dissolving the mixture in methanol, and heating and refluxing the mixture to obtain a coordination monomer M;

(2) dissolving the dried coordination monomer M in a solvent, adding a copper ion salt, and stirring for dissolving to obtain a mixed solution;

(3) placing the mixed solution obtained in the step (2) into a reaction kettle, and then placing the reaction kettle into an oven for standing reaction to obtain the anti-cancer drug;

in the above steps (1) to (3), the structural formula of the coordination monomer M is:

9. the method of claim 8, wherein the concentration of the compound of formula 2 in step (1) is 0.2-0.8 mol/L, the concentration of the compound of formula 3 in step (1) is 0.24-0.96 mol/L, the temperature of the heating reflux is 70-90 ℃ and the time is 3-5 hours.

10. The method of claim 8, wherein the solvent used in step (2) comprises 1-4 mL of analytically pure methanol, 0-3 mL of analytically pure DMF, and 3-9 mL of deionized water; the copper ion salt is copper nitrate trihydrate; the stirring time is 0.5 to 3 hours; the concentration of coordination monomer M in the mixed solution is 12.0 mg/mL-27.3 mg/mL, and the concentration of copper ions is 2.9 mg/mL-7.1 mg/mL;

and/or the standing reaction temperature in the step (3) is 65-90 ℃, and the reaction time is 48-96 hours.

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