Ring metal type organic gold (III) complex and preparation method thereof

Abstract

The invention provides a cyclometalated organic gold (III) complex and a preparation method thereof, and belongs to the technical field of medicines.The cyclometalated organic gold (III) complex is characterized in that a natural active molecule isoquinoline derivative is used as a CN ligand, and a carbon-metal sigma bond is introduced through a cyclometalation strategy to construct a main coordination structure; meanwhile, an anionic auxiliary ligand with a strong electron donating effect is introduced, so that the stability of a high-valence metal center is further enhanced, the oxidation-reduction stability of the high-valence metal center is improved, and the anti-tumor activity of the complex is improved. The invention has obvious inhibition on colon cancer cell proliferation, has high physiological stability, and provides technical support for development of new generation of antitumor metal drugs.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to a cyclic metal organogold (III) complex and its preparation method. Background Technology

[0002] In the field of clinical cancer treatment, chemotherapy remains an important means of combating malignant tumors. Although platinum-based drugs, represented by cisplatin, are widely used in chemotherapy for various malignant tumors, metal complexes have shown unique advantages in cancer chemotherapy due to their structural diversity and controllable biological effects. Among them, gold (III) complexes are particularly regarded as potential candidate drugs to overcome cisplatin resistance and toxic side effects. At the same time, in order to improve the stability of gold (III) complexes, the research focus has been on stabilizing the metal center through the rational design of ligands. The application of ring metallization strategy (introducing carbon-metal σ bonds) and strong field chelating ligands (such as C^N type donors) has been proven to be an effective way to enhance the redox stability of complexes. In addition, the research team has previously used isoquinoline alkaloids as ligands to verify their anticancer potential in platinum, ruthenium and other metal complexes, which has provided a foundation for the subsequent design of gold (III) complexes.

[0003] Currently, some gold (III) complexes have shown excellent antitumor activity in in vitro and in vivo models. In particular, cyclic metallized gold (III) complexes containing strong electron donor auxiliary ligands have further demonstrated excellent stability and significant antitumor effects. Based on previous research on isoquinoline alkaloid ligands, the concept of natural active molecular ligands and gold (III) centers has become an important practical direction for the development of gold (III) antitumor complexes.

[0004] However, the development of existing gold (III) complexes still faces key limitations: their inherent chemical instability is prominent, they are easily reduced under physiological conditions, and they have problems such as photosensitivity and kinetic instability, resulting in a lack of reliable in vivo antitumor efficacy pharmacological data for this type of compound, which seriously restricts further development; at the same time, the serious toxic side effects of traditional platinum drugs and the problem of tumor cells easily developing drug resistance have not been effectively solved, limiting their efficacy and application. Summary of the Invention

[0005] To overcome the shortcomings of the prior art, the purpose of this invention is to provide a cyclic metal organogold (III) complex and its preparation method. This invention significantly inhibits the proliferation of colon cancer cells and has high physiological stability, providing technical support for the development of a new generation of anti-tumor metal drugs.

[0006] To achieve the above objectives, the present invention provides the following solution: On one hand, the present invention provides a cyclic metal-type organometallic (III) complex, the structural formula of which is: ; Wherein, X^X is a bidentate anionic ligand of dithiocarbamate or a bidentate anionic ligand formed by the loss of one proton from a biguanide compound or a bidentate anionic ligand formed by the loss of two protons from an N,N'-diarylsulfonyl ethylenediamine compound; n is 0 or 1, Y - It is a chloride anion Cl - Or hexafluorophosphate anion F6P - .

[0007] Preferably, the structural formula of the dithiocarbamate bidentate anionic ligand is as follows: or ; Wherein, R is a C1~C4 alkyl group, including: methyl, ethyl, propyl, isopropyl and n-butyl; m values ​​include: 1, 2 and 3.

[0008] Preferably, the structure of the bidentate anionic ligand formed by the loss of one proton from the biguanide compound is as follows: ; Among them, R 1 It is a C1-C3 alkyl or phenyl; the C1-C3 alkyl includes: methyl, ethyl, n-propyl.

[0009] Preferably, the structural formula of the bidentate anionic ligand formed by the loss of two protons from the N,N'-diarylsulfonyl ethylenediamine compound is as follows: ; Where Ar is C6~C 10 The aryl group or the phenyl group substituted at the 4-position with a C1-C4 alkyl group, halogen, trifluoromethyl, nitro, or phenoxy group; the C6-C 10 The aryl group includes: phenyl, 2-naphthyl; the C1~C4 alkyl group includes: methyl, ethyl, propyl, isopropyl and tert-butyl; the halogen includes: fluorine, chlorine and bromine.

[0010] On the other hand, the present invention also provides a method for preparing the above-mentioned cyclic metal organogold(III) complex, comprising the following steps: S1. Under an inert atmosphere, protected from light and nitrogen, isoquinoline derivatives and mercuric acetate were dissolved in ethanol solvent and reacted at 110°C for 48 hours. Then, after cooling to room temperature, anhydrous lithium chloride methanol solution was added, and the reaction was continued at 100°C for 2 hours. After cooling, the first reaction solution was obtained. S2. Add deionized water to the first reaction solution, let it stand, filter and collect the filter cake, and purify the filter cake by silica gel column chromatography to obtain the purified compound. S3. Under an inert atmosphere, protected from light and nitrogen, the purified compound and potassium tetrachloroaurate were dissolved in a polar solvent and reacted at 110°C for 24 hours. After cooling to room temperature, a second reaction solution was obtained. The solution was filtered and the filter cake was collected to obtain the yellow intermediate (II). S4. The intermediate (II) is reacted with different anionic auxiliary ligands in polar solvents, filtered and the filter cake is collected. The filter cake is then separated and purified by column chromatography using an alkaline alumina packed column to obtain the corresponding cyclic metal organogold (III) complex.

[0011] Preferably, in S1, the structural formula of the isoquinoline derivative is: ; The molar ratio of the isoquinoline derivative, mercuric acetate, and anhydrous lithium chloride is 1:1:1 to 1:5:5.

[0012] Preferably, in step S3, the molar ratio of the purified compound to potassium tetrachloroaurate is 1:1 to 1:3; the volume ratio of the purified compound to the polar solvent is 1 mmol: 100 to 500 mL; the polar solvent is selected from one or more of N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, methanol, ethanol, ethylene glycol, ethylene glycol methyl ether, and water.

[0013] Preferably, in S4, the cyclic metal-type organogold(III) complex includes an organogold complex coordinated with dithiocarbamate anions, an organogold complex coordinated with biguanide anions, and an organogold complex coordinated with N,N'-diarylsulfonylethylenediamine anions. The specific preparation process is as follows: S41. The intermediate (II) and sodium dithiocarbamate were added to polar solvents respectively. The dithiocarbamate solution was added dropwise to the intermediate (II) solution while stirring. After stirring at room temperature for 12 hours, a saturated sodium hexafluorophosphate aqueous solution was added and allowed to stand for 2 hours. The filter cake was filtered and collected. The filter cake was then separated and purified by column chromatography using an alkaline alumina packed column to obtain the organogold complex with anion coordination of dithiocarbamate. S42. The intermediate (II), biguanide compound and potassium tert-butoxide are added to a polar solvent and stirred at room temperature for 12 hours. The mixture is then filtered to collect the filter cake, which is then purified by column chromatography using an alkaline alumina packed column to obtain the biguanide anion-coordinated organogold complex. S43. N,N'-diarylsulfonyl ethylenediamine compounds, triethylamine and deionized water were added to a polar solvent and reacted at 50°C for 30 minutes. Then intermediate (II) was added and the reaction was continued for 2 hours. Finally, the mixed reaction solution was filtered to collect the filter cake, and the filter cake was separated and purified by column chromatography using an alkaline alumina packed column to obtain the N,N'-diarylsulfonyl ethylenediamine anion-coordinated organogold complex. Specifically, the alkaline alumina packed column is used for column chromatography separation and purification of the filter cake, which employs alumina column chromatography. The alumina column chromatography uses a mixed solvent of dichloromethane and alcohols as the eluent. The volume ratio of dichloromethane to alcohols is 10~200:1. The alcohols include methanol and ethanol.

[0014] Preferably, the molar ratio of intermediate (II) to sodium dithiocarbamate is 1:1 to 1:5, the molar ratio of intermediate (II), biguanide compound and potassium tert-butoxide is 1:1:1 to 1:5:5, and the molar ratio of intermediate (II) to N,N'-diarylsulfonyl ethylenediamine compound is 1:1 to 1:5.

[0015] Preferably, in S41, the ratio of intermediate (II) to polar solvent is 1 mmol: 50~100 mL, and the ratio of sodium dithiocarbamate to polar solvent is 1 mmol: 50~100 mL. In S42, the ratio of intermediate (II) to polar solvent is 1 mmol: 50~250 mL; In S43, the ratio of intermediate (II) to polar solvent is 1 mmol: 50~250 mL, the volume ratio of triethylamine to deionized water is 1:1~1:3, and the ratio of N,N'-diarylsulfonyl ethylenediamine compound to triethylamine is 1 mmol: 1~10 mL.

[0016] According to specific embodiments provided by the present invention, the present invention discloses the following technical effects: This invention provides a series of novel cyclic organometallic gold complexes containing isoquinoline alkaloid derivatives and anionic ligands such as S^S or N^N. These complexes form a stable C^N main coordination framework through a cyclic metallization reaction between the isoquinoline alkaloid derivative and gold (III). The S^S or N^N anionic ligands then chelate with the gold (III) center. This strong chelation not only inhibits the reductive degradation of gold (III) under physiological conditions, thus enhancing physiological stability, but also synergistically regulates the activity of the gold (III) center through interligand electronic effects. Furthermore, experiments have demonstrated that these cyclic organometallic gold (III) complexes exhibit significant inhibitory activity against colon cancer cells, while also possessing high physiological stability, providing technical support for the development of next-generation antitumor metal drugs. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 This is a synthetic route diagram for the cyclic metal organogold(III) complexes of the present invention; wherein, Figure 1 Part (A) in the diagram is the synthetic route diagram for intermediate (II); Figure 1 Part (B) in the diagram is the synthetic route for cyclic metal organometallic (III) complexes; Figure 2 The high-performance liquid chromatography (HPLC) results of Au-1 prepared in Example 1 of this invention after exposure to physiological buffer solution for 0 h, 24 h and 48 h are shown. Figure 3 The high-performance liquid chromatography (HPLC) results of Au-2 prepared in Example 3 of this invention after exposure to physiological buffer solution for 0 h, 24 h and 48 h are shown. Figure 4 The high-performance liquid chromatography (HPLC) results of Au-3 prepared in Example 5 of this invention after exposure to physiological buffer solution for 0 h, 24 h and 48 h are shown. Figure 5 The high-performance liquid chromatography (HPLC) results of Au-4 prepared in Example 7 of this invention after exposure to physiological buffer solution for 0 h, 24 h and 48 h are shown. Figure 6 The image shows the high-performance liquid chromatography (HPLC) detection results of Au-5 prepared in Example 9 of this invention after exposure to physiological buffer solution for 0 h, 24 h, and 48 h. Detailed Implementation

[0019] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0020] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0021] like Figure 1As shown, this invention provides a synthetic route for cyclic metal organogold (III), including the synthetic route for intermediate (II), as follows. Figure 1 As shown in (A), the synthetic route for cyclic metal organogold(III) is as follows. Figure 1 As shown in (B) above. Based on the above, the preparation method of the cyclic metal organogold (III) of the present invention includes the following steps: S1. Under an inert atmosphere, protected from light and nitrogen, isoquinoline derivatives and mercuric acetate were dissolved in ethanol solvent and reacted at 110°C for 48 hours. Then, the mixture was cooled to room temperature and anhydrous lithium chloride methanol solution was added. The reaction was continued at 100°C for 2 hours. After cooling, the first reaction solution was obtained.

[0022] The structural formula of the isoquinoline derivative is as follows: ; Specifically, the molar ratio of isoquinoline derivative, mercuric acetate, and anhydrous lithium chloride is 1:1:1 to 1:5:5.

[0023] S2. Add deionized water to the first reaction solution, let it stand, filter and collect the filter cake, and purify the filter cake by silica gel column chromatography to obtain the purified compound.

[0024] S3. Under an inert atmosphere, protected from light and nitrogen, the purified compound and potassium tetrachloroaurate were dissolved in a polar solvent and reacted at 110°C for 24 hours. After cooling to room temperature, a second reaction solution was obtained. The solution was filtered and the filter cake was collected to obtain the yellow intermediate (II).

[0025] The molar ratio of the purified compound to potassium tetrachloroaurate is 1:1 to 1:3; the volume ratio of the purified compound to the polar solvent is 1 mmol: 100 to 500 mL; the polar solvent is selected from one or more of N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, methanol, ethanol, ethylene glycol, ethylene glycol methyl ether, and water.

[0026] S4. The intermediate (II) is reacted with different anionic auxiliary ligands in polar solvents, filtered and the filter cake is collected. The filter cake is then separated and purified by column chromatography using an alkaline alumina packed column to obtain the corresponding cyclic metal organogold (III) complex.

[0027] In the above preparation process, the cyclic metal organogold (III) complexes include organogold complexes coordinated with dithiocarbamate anions, organogold complexes coordinated with biguanide anions, and organogold complexes coordinated with N,N'-diarylsulfonylethylenediamine anions. The specific preparation process is as follows: S41. The intermediate (II) and sodium dithiocarbamate are added to a polar solvent respectively. The dithiocarbamate solution is added dropwise to the intermediate (II) solution while stirring. After stirring at room temperature for 12 hours, a saturated sodium hexafluorophosphate aqueous solution is added and allowed to stand for 2 hours. The filter cake is filtered and collected. The filter cake is then separated and purified by column chromatography using an alkaline alumina packed column to obtain the organogold complex with dithiocarbamate anion coordination.

[0028] S42. The intermediate (II), biguanide compound and potassium tert-butoxide are added to a polar solvent and stirred at room temperature for 12 hours. The mixture is then filtered to collect the filter cake, which is then purified by column chromatography using an alkaline alumina packed column to obtain the biguanide anion-coordinated organogold complex.

[0029] S43. N,N'-diarylsulfonyl ethylenediamine compounds, triethylamine, and deionized water were added to a polar solvent and reacted at 50°C for 30 minutes. Then, intermediate (II) was added, and the reaction was continued for 2 hours. Finally, the mixed reaction solution was filtered to collect the filter cake, and the filter cake was separated and purified by column chromatography using an alkaline alumina packed column to obtain the N,N'-diarylsulfonyl ethylenediamine anion-coordinated organogold complex.

[0030] Specifically, the alkaline alumina packed column is used for column chromatography separation and purification of the filter cake. The alumina column chromatography method uses a mixed solvent of dichloromethane and alcohols as the eluent; the volume ratio of dichloromethane to alcohols is 10~200:1; the alcohols include methanol and ethanol.

[0031] In the above specific preparation process, the molar ratio of intermediate (II) to sodium dithiocarbamate is 1:1 to 1:5, the molar ratio of intermediate (II), biguanide compound and potassium tert-butoxide is 1:1:1 to 1:5:5, and the molar ratio of intermediate (II) to N,N'-diarylsulfonyl ethylenediamine compound is 1:1 to 1:5.

[0032] In addition, in S41, the ratio of intermediate (II) to polar solvent is 1 mmol: 50~100 mL, and the ratio of sodium dithiocarbamate to polar solvent is 1 mmol: 50~100 mL. In S42, the ratio of intermediate (II) to polar solvent is 1 mmol: 50~250 mL; In S43, the ratio of intermediate (II) to polar solvent is 1 mmol: 50~250 mL, the volume ratio of triethylamine to deionized water is 1:1~1:3, and the ratio of N,N'-diarylsulfonyl ethylenediamine compound to triethylamine is 1 mmol: 1~10 mL.

[0033] The above content will be further explained below through specific implementation methods. The described embodiments are only some embodiments of the present invention.

[0034] It should be noted that the synthetic route of intermediate (II) involved in the following embodiments is as follows: .

[0035] Specifically, the preparation method of the above intermediate (II) includes the following steps: In a 200 mL round-bottom flask equipped with a magnetic stirrer, 1 g of isoquinoline derivative (4 mmol), 3.4 g of mercuric acetate (10 mmol), and 80 mL of ethanol were added sequentially. The mixture was heated to 110 °C and reacted for 48 hours under light-protected and nitrogen-protected conditions. After the reaction solution cooled to room temperature, anhydrous lithium chloride methanol solution was added, consisting of 0.84 g of anhydrous lithium chloride dissolved in 20 mL of methanol. The reaction was continued at 100 °C for 2 hours. After the reaction was complete, the reaction solution cooled to room temperature, 100 mL of deionized water was added to the flask, and the mixture was allowed to stand overnight. The mixture was then filtered, and the filter cake was collected. The filter cake was purified using silica gel column chromatography to obtain the purified compound. The eluent was a mixture of methanol and dichloromethane in a ratio of MeOH:DCM = 1:50.

[0036] In a 200 mL round-bottom flask equipped with a magnetic stirrer, 200 mg of the purified compound, 170 mg of potassium tetrachloroaurate (0.044 mmol), and 20 mL of acetonitrile were added sequentially. The mixture was reacted at 110 °C for 24 hours under light-protected and nitrogen-filled conditions. After the reaction was complete, the reaction solution was cooled to room temperature, filtered, collected, and dried to obtain intermediate (II), which could be directly used in the next reaction. High-resolution mass spectrometry characterization confirmed that the obtained compound was intermediate (II) of the present invention.

[0037] Example 1 This embodiment provides a cyclic metal organogold(III) complex, where the auxiliary ligand is... The target complex is referred to as Au-1. The preparation method of the target complex Au-1 includes the following steps: Under nitrogen protection, 30 mg of intermediate (II) (0.058 mmol) and 6 mL of methanol were added to a 50 mL Shrek reaction flask equipped with a magnetic stirrer. A methanol solution of sodium di-n-butyldithiocarbamate (13 mg of sodium di-n-butyldithiocarbamate dissolved in 3 mL of methanol) was slowly added dropwise while stirring. After the addition was complete, the mixture was stirred at room temperature for 12 hours. After the reaction was complete, the reaction solution was transferred to a 500 mL beaker, and 200 mL of saturated KPF6 aqueous solution was added. The mixture was allowed to stand for 2 hours. The filter cake was then collected by vacuum filtration and purified by column chromatography using an alkaline alumina-packed column to obtain Au-1 in 45% yield. The eluent used for purification was a mixture of ethyl acetate and hexane in a ratio of EA:hexane = 1:1.

[0038] The product obtained in Example 1 was characterized below, and the results are as follows: Nuclear magnetic resonance spectroscopy analysis yielded the following spectral data: 1H NMR spectrum: 1 H NMR (400 MHz, DMSO- d 6) δ 8.26 (d, J = 5.9 Hz, 1H), 8.12(d, J = 7.9 Hz, 1H), 8.00 (s, 1H), 7.80 (d, J = 6.3 Hz, 1H), 7.60 (s, 1H), 7.45 (t, J = 7.6 Hz, 1H), 7.33 (t, J = 7.4 Hz, 1H), 7.09 (d, J = 7.5 Hz, 1H),6.41 (s, 2H), 3.84 – 3.73 (m, 4H), 1.80 – 1.70 (m, 4H), 1.45 – 1.30 (m, 4H),0.97 (t, J = 7.4 Hz, 6H); Carbon NMR spectrum: 13 C NMR (100 MHz, DMSO- d6) δ 193.68, 160.64, 154.08, 152.16,151.31, 145.13, 139.97, 139.12, 132.29, 131.63, 129.49, 128.75, 123.40,123.26, 104.28, 102.33, 53.86, 52.29, 32.00, 29.88, 29.26, 28.95, 19.96,19.93; High-resolution mass spectrometry: HRMS (ESI) C 25 H 28 AuN2O2S2 + [M-F6P] + The calculated value is 649.1253, and the experimental value is 649.1249.

[0039] Based on the characterization results described above, the product obtained in Example 1 can be identified as the target complex Au-1, with the molecular formula C1. 25 H 28 AuN2O2S2PF6 has the following structure: .

[0040] Example 2 The preparation method of the target complex Au-1 in this embodiment differs from that in Example 1 in that methanol is replaced with ethylene glycol, while the other steps are the same. The yield obtained in this embodiment is 35%.

[0041] Furthermore, analysis of the product obtained in this embodiment using 1H NMR, 1C NMR, and high-resolution mass spectrometry confirmed that the product obtained in this embodiment is the target complex Au-1.

[0042] Example 3 This embodiment provides a cyclic organometallic gold (III) complex, wherein the auxiliary ligand is The target complex is referred to as Au-2. The preparation method of the target complex Au-2 includes the following steps: Under nitrogen protection, 30 mg of intermediate (II) (0.058 mmol) and 6 mL of methanol were added to a 50 mL Shrek reaction flask equipped with a magnetic stirrer. A methanol solution of sodium diethyldithiocarbamate trihydrate (13 mg sodium diethyldithiocarbamate dissolved in 3 mL of methanol) was slowly added dropwise while stirring. After the addition was complete, the mixture was stirred at room temperature for 12 hours. After the reaction was complete, the reaction solution was transferred to a 500 mL beaker, and 200 mL of saturated KPF6 aqueous solution was added. The mixture was allowed to stand for 2 hours. The filter cake was then collected by vacuum filtration and purified by column chromatography using an alkaline alumina-packed column to obtain Au-2 in a yield of 42%. The eluent used for purification was a mixture of ethyl acetate and n-hexane in a ratio of EA:hexane = 1:1.

[0043] The product obtained in Example 3 was characterized below, and the results are as follows: Nuclear magnetic resonance spectroscopy analysis yielded the following spectral data: 1H NMR spectrum: 1 H NMR (600 MHz, DMSO- d6 ) δ 8.19 (d, J = 6.3 Hz, 1H), 8.10 (d, J = 7.9 Hz, 1H), 8.00 (s, 1H), 7.76 (d, J = 6.2 Hz, 1H), 7.59 (s, 1H),7.48 – 7.37 (m, 1H), 7.33 – 7.25 (m, 1H), 7.01 (d, J = 7.5 Hz, 1H), 6.42 (s, 2H), 3.98 – 3.74 (m, 4H), 1.37 (dt, J = 11.6, 7.2 Hz, 6H); Carbon NMR spectrum: 13 C NMR (150 MHz, DMSO- d6 ) δ 191.93, 159.48, 159.47, 152.83,150.91, 150.01, 143.89, 138.87, 137.89, 131.03, 130.54, 128.25, 127.79,122.24, 103.20, 103.13, 101.29, 64.31, 47.84, 46.36, 14.57; High-resolution mass spectrometry: C 21 H 20 AuN2O2S2 + [M-F6P] +The calculated value is 593.0632, and the experimental value is 593.0620.

[0044] Based on the characterization results described above, the yellow solid product obtained in Example 3 can be identified as the target complex Au-2, with the molecular formula C2. 21 H 20 AuN2O2S2F6P has the following structure: .

[0045] Example 4 The preparation method of the target complex Au-2 in this embodiment differs from that in Example 3 in that methanol is replaced with ethylene glycol. All other steps are the same as in Example 3. The yield of the product obtained in this embodiment is 30%.

[0046] Furthermore, analysis of the yellow solid product obtained in this embodiment using 1H NMR, 1C NMR, and high-resolution mass spectrometry confirmed that the yellow solid product obtained in this embodiment is the target complex Au-2.

[0047] Example 5 This embodiment provides a cyclic organometallic gold (III) complex, wherein the auxiliary ligand is The target complex is referred to as Au-3. The preparation method for the target complex Au-3 is as follows: Under nitrogen protection, 0.058 mmol of intermediate (II) (30 mg) and 6 mL of methanol were added to a 50 mL Shrek reaction flask equipped with a magnetic stirrer. A methanol solution of sodium pyrrolidine dithiocarbamate (12 mg dissolved in 3 mL of methanol) was slowly added dropwise while stirring. After the addition was complete, the mixture was stirred at room temperature for 12 hours. After the reaction was complete, the reaction solution was transferred to a 500 mL beaker, and 200 mL of saturated KPF6 aqueous solution was added. The mixture was allowed to stand for 2 hours. The filter cake was then collected by vacuum filtration and purified by column chromatography using an alkaline alumina-packed column to obtain Au-3 in a yield of 39%. The eluent used for purification was a mixture of ethyl acetate and n-hexane in a ratio of EA:hexane = 1:1.

[0048] The product obtained in Example 5 was characterized below, and the results are as follows: Nuclear magnetic resonance spectroscopy analysis yielded the following spectral data: 1H NMR spectrum: 1 H NMR (400 MHz, DMSO- d 6) δ 8.23 ​​– 8.08 (m, 2H), 8.05 – 8.01(m, 1H), 7.79 (t, J= 3.5 Hz, 1H), 7.63 (d, J = 2.7 Hz, 1H), 7.42 (d, J = 8.1Hz, 1H), 7.32 – 7.22 (m, 1H), 6.99 – 6.92 (m, 1H), 6.43 (s, 2H), 3.85 (s,3H), 2.17 – 2.09 (m, 4H); Carbon NMR spectrum: 13 C NMR (100 MHz, DMSO- d 6) δ 187.93, 160.54, 154.10, 152.19,151.24, 145.00, 139.73, 139.13, 132.25, 131.61, 129.41, 128.50, 123.47,123.32, 104.30, 104.26, 102.33, 52.96, 52.29, 24.62, 24.31; High-resolution mass spectrometry: C 21 H 18 AuN2O2S2 + [M-F6P] + The calculated value is 591.0475, and the experimental value is 591.0469.

[0049] Based on the characterization results described above, the product obtained in Example 5 can be identified as the target complex Au-3, with the molecular formula C5. 21 H 18 AuN2O2S2PF6 has the following structure: .

[0050] Example 6 The preparation method of the target complex Au-3 in this embodiment differs from that in Example 5 in that methanol is replaced with ethylene glycol. All other steps are the same as in Example 5. The yield of the product obtained in this embodiment is 28%.

[0051] Furthermore, analysis of the product obtained in this embodiment using 1H NMR, 1C NMR, and high-resolution mass spectrometry confirmed that the product obtained in this embodiment is the target complex Au-3.

[0052] Example 7 This embodiment provides a cyclic organometallic gold (III) complex, wherein the auxiliary ligand is The target complex is referred to as Au-4. The preparation method for the target complex Au-4 is as follows: Under nitrogen protection, 50 mg of intermediate (II) (0.097 mmol), 200 mg of biguanide (1.45 mmol), 320 mg of potassium tert-butoxide (2.9 mmol), and 20 mL of methanol were added to a 50 mL Shrek reaction flask equipped with a magnetic stirrer. The mixture was stirred at room temperature for 12 hours. After the reaction was completed, the reaction solution was filtered and the filter cake was collected. The filter cake was purified by column chromatography using an alkaline alumina-packed column to obtain Au-4 with a yield of 35%. The eluent used for purification was a mixed solvent of methanol and dichloromethane in a ratio of MeOH:DCM = 1:30.

[0053] The product obtained in Example 7 was characterized below, and the results are as follows: Nuclear magnetic resonance spectroscopy analysis yielded the following spectral data: 1H NMR spectrum: 1 H NMR (600 MHz, DMSO- d 6 ) δ 8.61 (d, J = 6.4 Hz, 1H), 8.21 (dd, J = 7.8, 1.6 Hz, 1H), 8.17 (s, 1H), 7.93 (d, J = 6.5 Hz, 1H), 7.80 –7.74 (m, 1H), 7.64 (s, 1H), 7.54 – 7.45 (m, 2H), 7.04 (s, 1H), 6.58 (s, 2H), 6.42 (s, 2H), 6.34 (s, 2H), 6.16 (s, 1H); Carbon NMR spectrum: 13 C NMR (150 MHz, DMSO- d 6 ) δ 163.11, 159.95, 157.49, 154.14,151.96, 146.85, 145.31, 139.01, 137.32, 130.94, 128.88, 128.57, 123.10,121.78, 104.12, 103.95, 102.74; High-resolution mass spectrometry: C 18 H 16 AuN6O2 + [M-Cl] + The calculated value is 545.1000, and the experimental value is 545.0991.

[0054] Based on the characterization results described above, the yellow solid product obtained in Example 7 can be identified as the target complex Au-4, with the molecular formula C0. 18H 16 AuN6O2Cl ​​has the following structure: .

[0055] Example 8 The preparation method of the target complex Au-4 in this embodiment differs from that in Example 7 in that methanol is replaced with ethylene glycol. All other steps are the same as in Example 7. The yield of the product obtained in this embodiment is 26%.

[0056] Furthermore, analysis of the yellow solid product obtained in this embodiment using 1H NMR, 1C NMR, and high-resolution mass spectrometry confirmed that the yellow solid product obtained in this embodiment is the target complex Au-4.

[0057] Example 9 This embodiment provides a cyclic organometallic gold (III) complex, wherein the auxiliary ligand is The target complex is referred to as Au-5. The preparation method for the target complex Au-5 is as follows: Under nitrogen protection, 0.29 mmol of 100 mg N,N'-bis(p-toluenesulfonyl)ethylenediamine, 20 mL of methanol, and 2 mL of a 1:1 mixture of triethylamine and deionized water were added to a 50 mL Shrek reaction flask equipped with a magnetic stirrer. The reaction was carried out at 50 °C for 30 minutes. Subsequently, 0.097 mmol of 50 mg intermediate (II) was added, and the reaction was continued for 2 hours. After the reaction was completed, the reaction solution was filtered and the filter cake was collected. The filter cake was purified by column chromatography using an alkaline alumina-packed column to obtain Au-5 with a yield of 33%. The eluent used for purification was a mixture of methanol and dichloromethane in a ratio of MeOH:DCM = 1:30.

[0058] The product obtained in Example 9 was characterized below, and the results are as follows: Nuclear magnetic resonance spectroscopy analysis yielded the following spectral data: 1H NMR spectrum: 1 H NMR (400 MHz, DMSO- d 6) δ 8.76 (d, J = 6.6 Hz, 1H), 8.31(s, 1H), 8.23 ​​– 8.14 (m, 1H), 7.82 (d, J = 7.0 Hz, 2H), 7.77 (d, J = 8.3 Hz,2H), 7.69 (d, J = 8.2 Hz, 2H), 7.62 (s, 1H), 7.48 (td, J = 7.8, 1.3 Hz, 1H), 7.40 – 7.31 (m, 4H), 7.31 – 7.28 (m, 1H), 6.42 (s, 2H), 3.22 (t, J = 5.7 Hz, 2H), 2.95 (t, J = 5.6 Hz, 2H), 2.38 (d, J = 4.1 Hz, 6H); High-resolution mass spectrometry: C 32 H 29 N3O6S2Au + [M+H] + The calculated value is 812.1158, and the experimental value is 812.1124.

[0059] Based on the characterization results described above, the product obtained in Example 9 can be identified as the target complex Au-5, with the molecular formula C5. 32 H 28 N3O6S2Au has the following structure: .

[0060] The following experiments, conducted based on the above description, investigated the in vitro inhibitory activity of cyclic metallo-organogold (III) complexes against various cancer cell lines, specifically including: 1. Cell lines and cell culture The cell lines used in the experiment were: mouse triple-negative breast cancer cells (4T1 cells), human triple-negative breast cancer cells (MDA-MB-231 cells), human breast cancer cells (MCF-7), mouse colon cancer cells (CT-26), and mouse melanoma cells (B16-F10).

[0061] 4T1 cells were cultured in RPMI 1640 medium containing 10% fetal bovine serum, 1% sodium pyruvate, 100 U / mL penicillin, and 100 μg / mL streptomycin; MDA-MB-231 and MCF-7 cells were cultured in DMEM medium containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin; CT-26 and B16-F10 cells were cultured in RPMI 1640 medium containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin. All media were incubated at 37°C in a 5% CO2 incubator. The growth of 4T1, MDA-MB-231, MCF-7, CT-26, and B16-F10 cells was observed using an inverted microscope. Cells were passaged using 0.25% trypsin, and cells in the logarithmic growth phase were used for experiments.

[0062] 2. Preparation of the solution of the organometallic (III) complex of the analyte ring Five test ring-metal organogold complexes, Au-1, Au-2, Au-3, Au-4, and Au-5, were prepared according to the methods described in Examples 1, 3, 5, 7, and 9 of this invention, respectively. The products were all recrystallized twice from dichloromethane, and their purity was ≥95%. N,N-Dimethylformamide (DMF) stock solutions (2 mmol / L) of the test ring-metal organogold complexes Au-1, Au-2, Au-3, Au-4, and Au-5 were sequentially diluted to five concentration gradients (20 μmol / L, 10 μmol / L, 5 μmol / L, 2.5 μmol / L, 1.25 μmol / L, and 0.625 μmol / L) using RPMI-1640 or DMEM medium, with a final DMF concentration ≤1%, to test the inhibitory effects of different compounds on the growth of various cancer cells at different concentrations.

[0063] 3. Cell growth inhibition assay First, logarithmically growing 4T1 cells, MDA-MB-231 cells, MCF-7 cells, CT-26 cells, and B16-F10 cells were digested with trypsin and then cultured in medium containing 10% fetal bovine serum to prepare cell suspensions. 180 µL of each suspension was seeded into 96-well plates as experimental groups, ensuring a cell density of 1000-10000 cells / well. The edge wells were filled with sterile phosphate-buffered saline (PBS). Next, the cells were incubated in a 5% CO2, 37℃ incubator for 24 hours until 4T1 cells, MDA-MB-231 cells, MCF-7 cells, CT-26 cells and B16-F10 cells covered 60%~70% of the bottom of the experimental group wells in monolayer. 20µL of a solution of a certain concentration of the test ring metal-type organogold complex (Au-1, Au-2, Au-3, Au-4 and Au-5) was added to each well, with 5 replicates for each concentration. Next, the cells were incubated in a 5% CO2, 37°C incubator for 48 hours and observed under an inverted microscope. Then, add 20 µL of MTT solution (concentration of 5 mg / mL) to each well and incubate for another 4 h in an incubator; Subsequently, the culture was terminated, and the culture medium in the wells was carefully aspirated. 150 µL of dimethyl sulfoxide (DMSO) was added to each well to fully dissolve the formazan precipitate formed in 4T1, MDA-MB-231, MCF-7, CT-26, and B16-F10 cells after the addition of MTT solution. After vortexing to mix, the optical density of each well was measured using a microplate reader at a wavelength of 570 nm (reference wavelength 630 nm). Density (OD) values ​​were measured, and zeroing and control wells were set up. The zeroing wells included culture medium, 20 µL of MTT solution (concentration of 5 mg / mL) and 150 µL of DMSO. The control wells included 4T1 cells, MDA-MB-231 cells, MCF-7 cells, CT-26 cells and B16-F10 cells with the same number and condition as the experimental group, DMF at the same concentration as the experimental group, 20 µL of culture medium, 20 µL of MTT solution (concentration of 5 mg / mL) and 150 µL of DMSO.

[0064] Finally, the number of viable cells is determined based on the measured OD value; the higher the OD value, the stronger the cell activity. The inhibition rate of the compound on cancer cell growth is calculated using the formula: Cell growth inhibition rate = (OD value - OD value) / (Cell growth inhibition rate = OD value - OD value) 对照组 -OD 实验组 The inhibition rate data at five concentration gradients were fitted using the Bliss method to determine the half-maximal inhibitory concentration (IC50 value, in μmol / L) of the cyclic metallo-organogold complex against cancer cells.

[0065] Based on the above information, the test results are shown in Table 1.

[0066] Table 1. IC50 values ​​(μmol / L) of Au-1~Au-5 and cisplatin against five cancer cell lines.

[0067] The IC50 values ​​of cisplatin in Table 1 50The values ​​are derived from existing technology. Based on in vitro growth inhibition activity tests on cancer cells (4T1 cells, MDA-MB-231 cells, MCF-7 cells, CT-26 cells, and B16-F10 cells), the cyclic metalloid organogold complexes Au-1~Au-5 exhibited varying degrees of antitumor activity, with the best activity against CT-26 colon cancer cells (IC500). 50 The values ​​ranged from 1.03 μM to 9.98 μM. Au-1 showed the best inhibitory activity against all five cancer cell lines, with an IC50 value of [missing value]. 50 The values ​​range from 1.03 μM to 6.98 μM, and are all lower than the IC50 of cisplatin. 50 The value further proves that the cyclic metal organogold (III) complex of the present invention exhibits good in vitro antitumor activity against colon cancer cells and has potential medicinal value.

[0068] In addition, the present invention also conducted physiological stability experiments on cyclic organometallic gold (III) complexes, specifically including: 10 μL of Au-1 DMF stock solution (2 mmol / L) was added to 1 mL of TBS solution and incubated at 37 °C for 24 h and 48 h. After incubation, the changes were detected using a Waters Alliance e2695 HPLC system. The results are as follows: Figure 2 As shown.

[0069] 10 μL of Au-2 DMF stock solution (2 mmol / L) was added to 1 mL of TBS solution and incubated at 37 °C for 24 h and 48 h. After incubation, the changes were detected using a Waters Alliance e2695 HPLC system. The results are as follows: Figure 3 As shown.

[0070] 10 μL of Au-3 DMF stock solution (2 mmol / L) was added to 1 mL of TBS solution and incubated at 37 °C for 24 h and 48 h. After incubation, the changes were detected using a Waters Alliance e2695 HPLC system. The results are as follows: Figure 4 As shown.

[0071] 10 μL of Au-4 DMF stock solution (2 mmol / L) was added to 1 mL of TBS solution and incubated at 37 °C for 24 h and 48 h. After incubation, the changes were detected using a Waters Alliance e2695 HPLC system. The results are as follows: Figure 5 As shown.

[0072] 10 μL of Au-5 DMF stock solution (2 mmol / L) was added to 1 mL of TBS solution and incubated at 37 °C for 24 h and 48 h. After incubation, the changes were detected using a Waters Alliance e2695 HPLC system. The results are as follows: Figure 6 As shown.

[0073] like Figures 2 to 6 As shown, after 24 h and 48 h of treatment, no other obvious peaks of other components appeared in the cyclic organometallic gold (III) complexes of the present invention, indicating that the cyclic organometallic gold (Au-1, Au-2, Au-3, Au-4 and Au-5) complexes are stable in physiological buffer solutions, which further verifies the application of the cyclic organometallic gold (III) complexes provided by the present invention in the preparation of antitumor drugs.

[0074] Therefore, by employing the above-mentioned cyclic metal organogold (III) complex and its preparation method, the present invention significantly inhibits the proliferation of colon cancer cells and exhibits high physiological stability, providing technical support for the development of a new generation of anti-tumor metal drugs.

[0075] The various embodiments in this specification are described in a progressive manner, with each embodiment focusing on the differences from other embodiments. The same or similar parts between the various embodiments can be referred to each other.

[0076] This document uses specific examples to illustrate the principles and implementation methods of the present invention. The descriptions of the above embodiments are only for the purpose of helping to understand the method and core ideas of the present invention. Furthermore, those skilled in the art will recognize that, based on the ideas of the present invention, there will be changes in the specific implementation methods and application scope. Therefore, the content of this specification should not be construed as a limitation of the present invention.

Claims

1. A cyclic metal organogold(III) complex, characterized in that, The structural formula of the cyclic metal organometallic (III) complex is: ； Wherein, X^X is a bidentate anionic ligand of dithiocarbamate or a bidentate anionic ligand formed by the loss of one proton from a biguanide compound or a bidentate anionic ligand formed by the loss of two protons from an N,N'-diarylsulfonyl ethylenediamine compound; n is 0 or 1, Y - It is a chloride anion Cl - Or hexafluorophosphate anion F6P - .

2. The cyclic metal organometallic (III) complex according to claim 1, characterized in that, The structural formula of the dithiocarbamate bidentate anionic ligand is: or ; Wherein, R is a C1~C4 alkyl group, including: methyl, ethyl, propyl, isopropyl and n-butyl; m values ​​include: 1, 2 and 3.

3. The cyclic metal organometallic (III) complex according to claim 1, characterized in that, The structural formula of the bidentate anion ligand formed by the loss of one proton from the biguanide compound is as follows: ； Among them, R 1 It is a C1-C3 alkyl or phenyl; the C1-C3 alkyl includes: methyl, ethyl, n-propyl.

4. The cyclic metal organometallic (III) complex according to claim 1, characterized in that, The structural formula of the bidentate anionic ligand formed by the loss of two protons from the N,N'-diarylsulfonyl ethylenediamine compound is as follows: ； Where Ar is C6~C 10 The aryl group or the phenyl group substituted at the 4-position with a C1-C4 alkyl group, halogen, trifluoromethyl, nitro, or phenoxy group; the C6-C 10 The aryl group includes: phenyl, 2-naphthyl; the C1~C4 alkyl group includes: methyl, ethyl, propyl, isopropyl and tert-butyl; the halogen includes: fluorine, chlorine and bromine.

5. A method for preparing a cyclic metal organogold(III) complex according to any one of claims 1 to 4, characterized in that, Includes the following steps: S1. Under an inert atmosphere, protected from light and nitrogen, isoquinoline derivatives and mercuric acetate were dissolved in ethanol solvent and reacted at 110°C for 48 hours. Then, after cooling to room temperature, anhydrous lithium chloride methanol solution was added, and the reaction was continued at 100°C for 2 hours. After cooling, the first reaction solution was obtained. S2. Add deionized water to the first reaction solution, let it stand, filter and collect the filter cake, and purify the filter cake by silica gel column chromatography to obtain the purified compound. S3. Under an inert atmosphere, protected from light and nitrogen, the purified compound and potassium tetrachloroaurate were dissolved in a polar solvent and reacted at 110°C for 24 hours. After cooling to room temperature, a second reaction solution was obtained. The solution was filtered and the filter cake was collected to obtain the yellow intermediate (II). S4. The intermediate (II) is reacted with different anionic auxiliary ligands in polar solvents, filtered and the filter cake is collected. The filter cake is then separated and purified by column chromatography using an alkaline alumina packed column to obtain the corresponding cyclic metal organogold (III) complex.

6. The method for preparing a cyclic metal organogold(III) complex according to claim 5, characterized in that, In S1, the structural formula of the isoquinoline derivative is: ； The molar ratio of the isoquinoline derivative, mercuric acetate, and anhydrous lithium chloride is 1:1:1 to 1:5:

5.

7. The method for preparing a cyclic metal organogold(III) complex according to claim 5, characterized in that, In S3, the molar ratio of the purified compound to potassium tetrachloroaurate is 1:1 to 1:3; the volume ratio of the purified compound to the polar solvent is 1 mmol: 100 to 500 mL; the polar solvent is selected from one or more of N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, methanol, ethanol, ethylene glycol, ethylene glycol methyl ether, and water.

8. The method for preparing a cyclic metal organogold(III) complex according to claim 5, characterized in that, In S4, the cyclic metal organogold (III) complex includes organogold complexes coordinated with dithiocarbamate anions, organogold complexes coordinated with biguanide anions, and organogold complexes coordinated with N,N'-diarylsulfonylethylenediamine anions. The specific preparation process is as follows: S41. The intermediate (II) and sodium dithiocarbamate were added to polar solvents respectively. The dithiocarbamate solution was added dropwise to the intermediate (II) solution while stirring. After stirring at room temperature for 12 hours, a saturated sodium hexafluorophosphate aqueous solution was added and allowed to stand for 2 hours. The filter cake was filtered and collected. The filter cake was then separated and purified by column chromatography using an alkaline alumina packed column to obtain the organogold complex with anion coordination of dithiocarbamate. S42. The intermediate (II), biguanide compound and potassium tert-butoxide are added to a polar solvent and stirred at room temperature for 12 hours. The mixture is then filtered to collect the filter cake, which is then purified by column chromatography using an alkaline alumina packed column to obtain the biguanide anion-coordinated organogold complex. S43. N,N'-diarylsulfonyl ethylenediamine compounds, triethylamine and deionized water were added to a polar solvent and reacted at 50°C for 30 minutes. Then intermediate (II) was added and the reaction was continued for 2 hours. Finally, the mixed reaction solution was filtered to collect the filter cake, and the filter cake was separated and purified by column chromatography using an alkaline alumina packed column to obtain the N,N'-diarylsulfonyl ethylenediamine anion-coordinated organogold complex. Specifically, the alkaline alumina packed column is used for column chromatography separation and purification of the filter cake, which employs alumina column chromatography. The alumina column chromatography uses a mixed solvent of dichloromethane and alcohols as the eluent. The volume ratio of dichloromethane to alcohols is 10~200:

1. The alcohols include methanol and ethanol.

9. The method for preparing a cyclic metal organogold(III) complex according to claim 8, characterized in that, The molar ratio of intermediate (II) to sodium dithiocarbamate is 1:1 to 1:5, the molar ratio of intermediate (II), biguanide compound and potassium tert-butoxide is 1:1:1 to 1:5:5, and the molar ratio of intermediate (II) to N,N'-diarylsulfonyl ethylenediamine compound is 1:1 to 1:

5.

10. The method for preparing a cyclic metal organogold(III) complex according to claim 8, characterized in that, In S41, the ratio of intermediate (II) to polar solvent is 1 mmol: 50~100 mL, and the ratio of sodium dithiocarbamate to polar solvent is 1 mmol: 50~100 mL. In S42, the ratio of intermediate (II) to polar solvent is 1 mmol: 50~250 mL; In S43, the ratio of intermediate (II) to polar solvent is 1 mmol: 50~250 mL, the volume ratio of triethylamine to deionized water is 1:1~1:3, and the ratio of N,N'-diarylsulfonyl ethylenediamine compound to triethylamine is 1 mmol: 1~10 mL.

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