CN110128452B

CN110128452B - Gold complex and synthesis method and application thereof

Info

Publication number: CN110128452B
Application number: CN201910372292.4A
Authority: CN
Inventors: 黄克斌
Original assignee: Guangxi Normal University
Current assignee: Guangxi Normal University
Priority date: 2018-02-05
Filing date: 2018-02-05
Publication date: 2020-12-01
Anticipated expiration: 2038-02-05
Also published as: CN108148080B; CN108148080A; CN110128452A

Abstract

The invention discloses a gold complex and a synthesis method and application thereof. The synthesis method of the gold complex comprises the following steps: placing chloroauric acid tetrahydrate or chloroauric acid salt and phenethylamine ligand in an organic solvent, coordinating under the condition of keeping out of the sun, cooling the reactant, standing for crystallization, and collecting crystals to obtain the chlorauric acid tetrahydrate or chloroauric acid salt and phenethylamine ligand; wherein the chloraurate is sodium or potassium chloraurate, and the phenylethylamine ligand is piperonylethylamine. Experiments of the applicant show that the complex has good proliferation inhibition activity and drug resistance to certain tumor cell strains, has low toxicity to human normal cells HL-7702, and also has high physiological stability. The complex structure of the invention is shown as the following formula (I):

Description

Gold complex and synthesis method and application thereof

the application is a divisional application of 'metal organic gold (III) complex and a synthesis method and application thereof', and the application date of the original application is as follows: and on 5 days 2 months in 2018, the application numbers are as follows: 201810111445.5, title of the invention: a metal organic gold (III) complex and a synthesis method and application thereof.

Technical Field

The invention relates to the technical field of medicines, in particular to a metal organic gold (III) complex and a synthesis method and application thereof.

Background

Cisplatin and its analogs carboplatin and oxaliplatin have been successfully used to treat various solid tumors, but severe drug resistance and side effects limit the clinical utility of these platinum drugs, and thus metal complexes with stronger anticancer activity and lower side effects are of interest to numerous scholars. Some of the non-platinum metal complexes show high-efficiency and low-toxicity drug design attributes, and some of the gold complexes also show anticancer mechanisms which are obviously different from those of platinum drugs, however, the low physiological stability of the general gold complexes prevents the in vivo therapeutic effect of the gold complexes.

1,2,3, 4-Tetrahydroisoquinoline (THIQ) is a special alkaloid compound, widely exists in natural products, and has certain bioactivity. At present, metallo compounds of (iso) quinoline and various biological activities thereof have been widely researched, but no relevant report has been found on the synthesis and activity research of inserting a metal atom into an N-heterocyclic structure of THIQ to replace one carbon atom.

Disclosure of Invention

The invention aims to solve the technical problem of providing a metal organic gold (III) complex with a novel structure and a synthesis method and application thereof.

The metal organic gold (III) complex is a compound shown as the following formula (I) or a pharmaceutically acceptable salt thereof:

wherein R is₁Represents OMe or OCH₂O，R₂Represents OMe or OCH₂O。

The synthesis method of the metal organic gold (III) complex comprises the following steps: placing chloroauric acid tetrahydrate or chloroauric acid salt and phenethylamine ligand in an organic solvent, coordinating under the condition of keeping out of the sun, cooling the reactant, standing for crystallization, and collecting crystals to obtain a target product; wherein the chloroaurate is sodium chloroaurate or potassium chloroaurate, and the phenylethylamine ligand is 3, 4-dimethoxyphenylethylamine or piperonylethylamine.

In the above synthesis method, the ratio of the amounts of the chloroauric acid tetrahydrate or the chloroauric acid salt and the phenylethylamine ligand is a stoichiometric ratio, and in actual operation, the molar ratio of the chloroauric acid tetrahydrate or the chloroauric acid salt to the phenylethylamine ligand may be generally 1 to 5: 1.

in the above synthesis method, the organic solvent may be any one or a combination of two or more selected from dichloromethane, acetone, ethanol, methanol and chloroform. When the organic solvent is a combination of two or more of the above, the ratio of the organic solvents may be any ratio. The organic solvent is preferably used before use

The molecular sieve dehydrates, thereby being more beneficial to the reaction. The amount of the organic solvent can be determined according to the need, and is usually the raw material capable of dissolving and participating in the reaction, specifically, the total amount of the organic solvent used for all the raw materials is usually 10 to 150mL, preferably 15 to 50mL, calculated by taking 0.1mmol of chloroauric acid tetrahydrate or chloroauric acid salt and 0.1mmol of phenethylamine ligand as the reference. When the amount of the organic solvent added is large, after the reaction is stopped, most of the organic solvent is removed by a method such as reduced pressure distillation (usually, the removal accounts for 80 to 95% of the amount of the organic solvent added), and then the reaction solution is cooled and left to stand to precipitate the target product.

In the above synthesis method, the reaction may be carried out with or without heating. When the reaction is carried out under heating, the reaction temperature is lower than the boiling point temperature of the organic solvent, and preferably the reaction is carried out at 50 to 80 ℃. Whether the reaction is complete or not can be followed by Thin Layer Chromatography (TLC). After the reaction is completed, the reaction product is preferably filtered, cooled and kept stand for crystallization. The standing time is usually 1 to 5 days.

The yield of the metal organic gold (III) complex prepared by the method is more than 40%, specifically 40-90%. The prepared metal organic gold (III) complex can be further purified by a recrystallization method, and the solvent used in recrystallization is the same as the organic solvent.

The invention also comprises the application of the metal organic gold (III) complex or the pharmaceutically acceptable salt thereof in preparing antitumor drugs.

The invention further includes a pharmaceutical composition comprising a therapeutically effective amount of the above-described metal-organic gold (III) complex or a pharmaceutically acceptable salt thereof.

Compared with the prior art, the invention provides two metal organic gold (III) complexes with novel structures and a synthesis method and application thereof, and experiments of the applicant show that the metal organic gold (III) complexes have good proliferation inhibition activity and drug resistance to certain tumor cell strains, have low toxicity to human normal cells HL-7702 and have high physiological stability. The complex has the advantages of simple synthesis method, mild reaction conditions, cheap and easily-obtained reaction raw materials and low cost.

Drawings

FIG. 1 is a single crystal structural view of a final product obtained in example 1 of the present invention;

FIG. 2 is a single crystal structural view of a final product obtained in example 6 of the present invention;

FIG. 3 is a graph showing HPLC test results of Cyc-Au-1 prepared by the method of example 1 of the present invention in physiological buffer solution for 24h and 48 h;

FIG. 4 is a graph showing the results of HPLC analysis of Cyc-Au-2 prepared by the method of example 6 in physiological buffer solution for 24h and 48 h.

Detailed Description

The present invention will be better understood from the following detailed description of specific examples, which should not be construed as limiting the scope of the present invention.

Example 1: synthesis of target product Cyc-Au-1

Taking KAuCl₄(0.2mmol) and 3, 4-dimethoxyphenethylamine (0.2mmol) are placed in a container, then 30mL of mixed solvent composed of dichloromethane and methanol (the volume ratio of dichloromethane to methanol is 1: 1) is added, the obtained mixture is refluxed and stirred for reaction for 24 hours under the condition of keeping out of the sun, the obtained reactant is filtered, filtrate is collected and kept stand, yellow crystals are separated out, the crystals are collected and dried in vacuum, and yellow solid products are obtained with the yield of 55 percent.

The product obtained in this example was characterized:

1) and (3) performing nuclear magnetic hydrogen spectrum analysis, wherein the obtained spectral data are as follows:

nuclear magnetic hydrogen spectrum:¹H NMR(400MHz,DMSO-d₆)6.84(d,J＝8.1Hz,1H),6.70(dd,J＝8.1,1.6Hz,1H),3.74(s,3H),3.72(s,3H),2.81–2.67(t,J＝5.4Hz 2H),2.57(t,J＝7.2Hz,2H)。

2) electrospray mass spectrometry: m/z 490.4[ M-Cl + DMSO ]]⁺。

3) X-ray single crystal diffraction analysis:

the obtained blackish green plate-like crystal was subjected to single crystal X-ray analysis by an agilent SuperNova single crystal diffractometer, and its crystallographic data are shown in table 1 below, and the crystal structure of the obtained product is shown in fig. 1.

Table 1: crystallographic data of Cyc-Au-1 and Cyc-Au-2

Therefore, the yellow solid product obtained in the example can be determined to be the target product Cyc-Au-1 with the molecular formula of C₁₀H₁₄NO₂Cl₂Au, the structure of which is shown as the following formula:

example 2: synthesis of target product Cyc-Au-1

Example 1 was repeated except that: chloroform was used instead of the mixed solvent consisting of dichloromethane and methanol. The yield was 58%.

And (3) performing nuclear magnetic hydrogen spectrum, electrospray mass spectrum and X-ray single crystal diffraction analysis on the product obtained in the embodiment, and determining that the product obtained in the embodiment is the target product Cyc-Au-1.

Example 3: synthesis of target product Cyc-Au-1

Example 1 was repeated except that: the reaction is carried out at 25 ℃ for 48 h. The yield was 46%.

Example 4: synthesis of target product Cyc-Au-1

Example 1 was repeated except that: with HAuCl₄·4H₂O instead of KAuCl₄. The yield was 59%.

Example 5: synthesis of target product Cyc-Au-1

Example 1 was repeated except that: with NaAuCl₄Substitute for KAuCl₄. The yield was 58%.

Example 6: synthesis of target product Cyc-Au-2

Taking NaAuCl₄(0.2mmol) and 3, 4-dimethyleneoxyphenylethylamine (0.2mmol) are placed in a container, then 30mL of mixed solvent composed of dichloromethane and methanol (the volume ratio of dichloromethane to methanol is 1: 1) is added, the obtained mixture is refluxed and stirred for reaction for 24 hours under the condition of keeping out of the light, the obtained reactant is filtered, filtrate is collected and kept stand, yellow crystals are separated out, the crystals are collected and dried in vacuum, and the yellow solid product is obtained with the yield of 43 percent.

The product obtained in this example was characterized:

nuclear magnetic hydrogen spectrum:¹H NMR(400MHz,DMSO-d₆)6.81(dd,J＝7.7,3.6Hz,1H),6.65(dd,J＝7.9,1.7Hz,1H),5.96(s,2H),2.73(t,J＝7.2Hz,1H),2.56(t,J＝7.2Hz,1H)。

2) electrospray mass spectrometry: m/z 474.9[ M-Cl + DMSO]⁺。

3) X-ray single crystal diffraction analysis:

the obtained blackish green plate-like crystal was subjected to single crystal X-ray analysis by an agilent SuperNova single crystal diffractometer, and the crystallographic data thereof are shown in table 1 above, and the crystal structure of the obtained product is shown in fig. 2.

Therefore, the yellow solid product obtained in the example can be determined to be the target product Cyc-Au-2 with the molecular formula of C₉H₁₀NO₂Cl₂Au, the structure of which is shown as the following formula:

example 7: synthesis of target product Cyc-Au-2

Example 6 was repeated except that: replacing a mixed solvent consisting of dichloromethane and methanol with a mixture of ethanol and chloroform, wherein the volume ratio of ethanol to chloroform is 3: 1. the yield was 53%.

And (3) performing nuclear magnetic hydrogen spectrum, electrospray mass spectrum and X-ray single crystal diffraction analysis on the product obtained in the embodiment, and determining that the product obtained in the embodiment is the target product Cyc-Au-2.

Example 8: synthesis of target product Cyc-Au-2

Example 6 was repeated except that: the mixed solvent composed of dichloromethane and methanol is replaced by acetone, and the reaction is carried out at 40 ℃ for 24 h. The yield was 43%.

Example 9: synthesis of target product Cyc-Au-2

Example 6 was repeated except that: with HAuCl₄·4H₂O instead of KAuCl₄. The yield was 45%.

Example 10: synthesis of target product Cyc-Au-2

Example 6 was repeated except that: with KAuCl₄Replacing NaAuCl₄. The yield is 48 percent。

Experimental example 1: the complex of the invention is used for carrying out in vitro inhibitory activity experiments on various human tumor strains:

1. cell lines and cell cultures

The experiment selects human liver cancer cell BEL-7404, bladder cancer cell HepG-2, ovarian cancer cell SK-OV-3, lung cancer cell A549, lung cancer cis-platinum resistant cell A549/CDDP, colon cancer cell HCT116 and human normal liver cell HL-7702.

All cell lines were cultured in RPMI-1640 or DMEM medium containing 10% fetal bovine serum, 100U/mL penicillin, and 100U/mL streptomycin, and placed at 37 deg.C with 5% CO by volume₂Culturing in an incubator. The growth of the cells was observed by inverted microscope, and the cells were passed through digestion with 0.25% trypsin and used in the experiment in the logarithmic phase.

2. Preparation of test Compounds

The Cyc-Au-1 and Cyc-Au-2 are obtained by recrystallizing products obtained by the methods of the embodiment 1 and the embodiment 6 of the invention for 2 times by using dichloromethane, the purity of the products is more than or equal to 95 percent, DMSO stock solution (the concentration is 0.002mol/L) is sequentially diluted into five concentration gradients which are respectively 40, 20, 10, 5 and 2.5 mu mol/L by RPMI-1640 or DMEM culture medium, and the final concentration of the cosolvent DMSO is less than or equal to 1 percent. Respectively testing the proliferation inhibition degree of the target product to various tumor cells under different gradient concentrations for fitting and calculating the half inhibition concentration, namely IC₅₀The value is obtained. All tests were repeated at least three times independently.

3. Cell growth inhibition assay (MTT method)

(1) Taking tumor cells in logarithmic growth phase, digesting by trypsin, preparing cell suspension by using culture solution containing 10% fetal calf serum, inoculating 180 mu L of the cell suspension into a 96-well culture plate, and enabling the cell density to be detected to reach 1000-10000 cells/well (the edge hole is filled with sterile PBS);

(2)5％CO₂incubating for 24 hours at 37 ℃ until cell monolayers are paved to 60-70% of the bottom of the hole, and adding a certain amount of the suspension into each hole20 mu L of medicine with concentration gradients, and each concentration gradient is provided with 5 compound holes;

(3)5％CO₂incubating for 48h at 37 ℃, and observing under an inverted microscope;

(4) mu.L of MTT solution (5mg/mL) was added to each well and incubation was continued for 4h

(5) Terminating the culture, carefully removing the culture solution in the wells, adding 150. mu.L DMSO into each well to sufficiently dissolve formazan precipitate, mixing uniformly by using an oscillator, and measuring the optical density of each well at a wavelength of 570nm and a reference wavelength of 630nm by using an enzyme labeling instrument;

(6) setting zero adjusting holes (culture medium, MTT and DMSO) and control holes (cells, drug dissolving medium with the same concentration, culture solution, MTT and DMSO);

(7) the number of living cells was judged from the measured optical density values (OD values), and the larger the OD value, the stronger the cell activity. Using the formula:

and (4) calculating the inhibition rate of the compound on the growth of tumor cells. The test results are shown in table 2 below:

table 2: growth inhibition Rate (%) of Cyc-Au-1 and Cyc-Au-2 against 6 tumor cell lines and 1 Normal cell line

Note: human liver cancer cell BEL-7404, bladder cancer cell HepG-2, ovarian cancer cell SK-OV-3, lung cancer cell A549, lung cancer cis-platinum resistant cell A549/CDDP, colon cancer cell HCT116 and human normal liver cell HL-7702, wherein the primary screening concentration of the selected compounds is 20 mu mol/L.

Further, inhibition rate data of five concentration gradients are fitted through Bliss software, and half Inhibition Concentration (IC) of the product on different tumor strains is calculated₅₀Values in μmol/L), the results are shown in table 3 below:

table 3: Cyc-Au-1 and Cyc-Au-2 pairs 6 tumor cell strains and one normal strainIC of cell lines₅₀Value (μ M)

From the results of in vitro antitumor activity tests, the complexes Cyc-Au-1 and Cyc-Au-2 both have broad-spectrum anticancer activity, and meanwhile, the two gold complexes show lower cytotoxic activity on normal human hepatocytes HL-7702. It can be seen that complexes Cyc-Au-1 and Cyc-Au-2 show a certain degree of selectivity on cancer cells. When two gold complexes are compared, Cyc-Au-2 has higher tumor cell inhibition activity. Compared with the clinical anticancer drug cisplatin, Cyc-Au-2 also shows better tumor growth inhibition activity and lower cytotoxicity to normal liver cells HL-7702 on the whole, so Cyc-Au-2 has better cytotoxicity selectivity than cisplatin. Meanwhile, in terms of the drug resistance to the cis-platinum drug-resistant strain A549/DDP, the drug resistance factors RF of Cyc-Au-1 and Cyc-Au-2 to the A549/DDP are respectively 1.42 and 1.15, while the drug resistance factor of the cis-platinum is 7.12, and the results show that both Cyc-Au-1 and Cyc-Au-2 have lower drug resistance than the cis-platinum.

In conclusion, the metal organic gold (III) complex compound provided by the invention generally shows a certain in vitro anti-tumor activity, has obvious toxicity selectivity and anti-drug resistance effect on cancer cells, has good potential medicinal value, and is expected to be used for preparing various anti-tumor medicaments.

Experimental example 2: biostability experiments for the complexes of the invention

(1) A10. mu.L DMSO stock solution (2X 10) of Cyc-Au-1 (prepared as described in example 1 of the invention) was taken^-3mol/L) were added to the 1ml PBS solution, incubated at 37 ℃ for 24h and 48h, and the change was detected by Waters 2950HPLC after the incubation, the results are shown in FIG. 3.

(2) A10. mu.L DMSO stock solution (2X 10) of Cyc-Au-2 (prepared as described in example 6 of the invention) was taken^-3mol/L) was added to the 1ml PBS solution, incubated at 37 ℃ for 24h and 48h, and the change was detected by Waters 2950HPLC after the incubation, the results of which are shown in FIG. 4.

As can be seen from FIGS. 3 and 4, no other component peaks were evident after the two complexes were exposed for 24 and 48 hours, respectively, indicating that the two complexes were stable in physiological buffer solutions.

Claims

1. A compound of the formula:

2. a method of synthesizing the compound of claim 1, wherein: the method mainly comprises the following steps: placing chloroauric acid tetrahydrate or chloroauric acid salt and phenethylamine ligand in an organic solvent, coordinating under the condition of keeping out of the sun, cooling the reactant, standing for crystallization, and collecting crystals to obtain a target product; wherein the chloraurate is sodium or potassium chloraurate, and the phenylethylamine ligand is piperonylethylamine.

3. The method of synthesis according to claim 2, characterized in that: the organic solvent is selected from any one or the combination of more than two of dichloromethane, acetone, ethanol, methanol and chloroform.

4. The method of synthesis according to claim 2, characterized in that: when the reaction is carried out under heating, the reaction temperature is lower than the boiling temperature of the organic solvent.

5. The method of synthesis according to claim 4, characterized in that: the reaction is carried out at 50-80 ℃.

6. The method of synthesis according to any one of claims 2-5, characterized in that: further comprises a purification step: specifically, the prepared target product is recrystallized to obtain a purified target product.

7. The use of a compound of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of an anti-neoplastic drug.

8. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1 or a pharmaceutically acceptable salt thereof.