A zinc complex, its preparation method and application
By preparing zinc complexes with good biosafety as sound sensitizers, the biosafety and metabolic problems of sound sensitizers in existing acoustic dynamic treatments are solved, and the non-invasive therapeutic effect on deep tumors is achieved.
Patent Information
- Application Number
- CN202311447833.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-11-01
- Publication Date
- 2025-08-01
- Estimated Expiration
- 2043-11-01
AI Technical Summary
The lack of sound-sensitive agents with good biosafety, clear structure and easy metabolism in existing acoustic dynamics therapy limits its application in deep tumor treatment.
A zinc complex was developed to prepare zinc complexes through specific chemical synthesis methods, which act as a sound-sensitive agent, and generate singlet oxygen under ultrasound activation to kill cancer cells, with good biosafety and structural clarity.
Zinc complexes can produce a large amount of singlet oxygen under ultrasound conditions, destroy the redox balance of cancer cells, show excellent anti-tumor effects, and have no obvious toxicity to normal cells. They are suitable for non-invasive treatment of deep tumors.
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Figure CN117603233B_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of pharmaceutical chemistry, and particularly relates to a zinc complex and its preparation method and application. Background Art
[0002] In recent years, many studies have reported the development of novel non-invasive treatment methods for various diseases including cancer. One such method is photodynamic therapy (PDT), which can generate reactive oxygen species (ROS) by transferring the energy of light to a photosensitizer. However, the penetration ability of light, including near-infrared light, is poor, which limits the future development of photodynamic therapy. To overcome the limitation of light penetration, ultrasound (US), a deeply penetrating mechanical wave, has become a key diagnostic technology in hospitals and clinics. US can not only penetrate deeper tissue layers but also reduce damage to surrounding normal tissues. At the same time, US can also activate sonosensitizers and generate reactive oxygen species (ROS), such as singlet oxygen, thereby killing malignant tumors or weakening their proliferation potential, namely sonodynamic therapy (SDT). SDT has the characteristics of deep penetration, non-invasiveness, high biosafety, and low cost, and is suitable for non-invasive treatment methods for various deep tumors. So far, a series of ultrasound-active nanomaterials with good physical and chemical properties have been developed, but uncertain composition and poor biosafety limit their extensive therapeutic applications. Therefore, it is necessary to develop sonosensitizers with good biosafety, clear structure, and easy metabolism to promote the development of sonodynamic therapy for tumors. Summary of the Invention
[0003] In view of the above deficiencies of the prior art, the present invention provides a zinc complex and its preparation method and application, aiming to solve the problem of the lack of sonosensitizers with good biosafety, clear structure, and easy metabolism in existing sonodynamic therapy.
[0004] Specifically, the technical solution of the present invention is as follows:
[0005] The present invention provides a zinc complex, and the chemical structural formula of the zinc complex is shown as follows:
[0006]
[0007] For the zinc complex, in an aqueous solution with a DMSO volume fraction of 10%, the absorption wavelength of the zinc complex is 445 nm.
[0008] For the zinc complex, in an aqueous solution with a DMSO volume fraction of 10%, the emission wavelength of the zinc complex is about 600 nm.
[0009] The present invention provides a preparation method of the zinc complex, comprising the steps:
[0010] S1. Add 4-methoxy-N-(4-methoxyphenyl)-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxolan-2-yl)phenyl)aniline and 3,8-dibromo-1,10-phenanthroline to the first solvent, add Pd(PPh)4 and K2CO3, and heat under reflux in an inert atmosphere. After the reaction, ligand TP is obtained.
[0011] S2. Add zinc acetate and the ligand TP to the second solvent, and heat under reflux in an inert atmosphere to obtain a crude solid. After purifying the crude solid, the zinc complex is obtained.
[0012] The method for preparing the zinc complex, wherein the molar ratio of 4-methoxy-N-(4-methoxyphenyl)-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxolan-2-yl)phenyl)aniline to 3,8-dibromo-1,10-phenanthroline is 2.2:1.
[0013] The method for preparing the zinc complex, wherein the first solvent includes tetrahydrofuran and water, and the volume ratio of tetrahydrofuran to water is 1:1.
[0014] The method for preparing the zinc complex, wherein the molar ratio of zinc acetate to the ligand TP is 1:1.
[0015] The method for preparing the zinc complex, wherein the second solvent includes toluene and methanol, and the volume ratio of toluene to methanol is 1:1.
[0016] The method for preparing the zinc complex, wherein, optionally, in step S1, the temperature of the heating under reflux is 75 °C.
[0017] Optionally, in step S2, the temperature of the heating under reflux is 90 °C.
[0018] The present invention also provides an application of the zinc complex in preparing a drug for sonodynamic therapy of tumors.
[0019] Compared with the prior art, the present invention has the following beneficial effects:
[0020] The present invention provides a zinc complex, a preparation method and an application thereof. The zinc complex has good biosafety. In the absence of ultrasound, it cannot generate singlet oxygen 1 O2, and has extremely low killing ability to cells; under ultrasound irradiation at 1.0 MHz, 3 W cm -2 , 10% duty cycle, it can generate a large amount of singlet oxygen 1O2, thereby disrupting the intracellular redox balance and leading to the death of cancer cells. The anti-tumor activity of the zinc complex was evaluated using mouse breast cancer cells (4T1). In the absence of ultrasound, the zinc complex had no obvious cytotoxicity to 4T1 cells, while under ultrasound conditions, it showed good anti-tumor activity against 4T1 cells. The IC 50 value of the zinc complex against 4T1 cells could reach 2.45 μM, showing excellent anti-tumor effects and promising for sonodynamic therapy of tumors. BRIEF DESCRIPTION OF THE DRAWINGS
[0021] Figure 1 1H NMR spectrum of ligand TP in the examples of the present invention;
[0022] Figure 2 1H NMR spectrum of zinc complex Zn-TP in the examples of the present invention;
[0023] Figure 3 UV-Vis absorption spectrum of zinc complex Zn-TP in the examples of the present invention;
[0024] Figure 4 Fluorescence emission spectrum of zinc complex Zn-TP in the examples of the present invention;
[0025] Figure 5 UV-Vis absorption spectrum of zinc complex Zn-TP using 9,10-diphenylanthracene (DPA) as a singlet oxygen probe under ultrasound conditions in the examples of the present invention;
[0026] Figure 6 Effect of zinc complex Zn-TP on the survival rate of mouse melanoma cells under dark and ultrasound conditions in the examples of the present invention. DETAILED DESCRIPTION OF THE INVENTION
[0027] The present invention provides a zinc complex, its preparation method and application. To make the purpose, technical solution and effects of the present invention clearer and more definite, the present invention is further described in detail below. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the protection scope of the present invention.
[0028] An embodiment of the present invention provides a zinc complex, and its chemical structural formula is as follows:
[0029]
[0030] In the structure of the above zinc complex, the zinc element belongs to transition metals. Compared with heavy metals, it has less biological dark toxicity and side effects. Therefore, as a sonosensitizer, this zinc complex has the advantages of good biological safety, clear structure and easy metabolism, and generates 1O2 can effectively kill tumor cells and achieve excellent anti-tumor effects.
[0031] In one embodiment, in an aqueous solution with a DMSO volume fraction of 10%, the absorption wavelength of the zinc complex is 445 nm.
[0032] In one embodiment, in an aqueous solution with a DMSO volume fraction of 10%, the emission wavelength of the zinc complex is approximately 600 nm.
[0033] The embodiments of the present invention provide a preparation method of the zinc complex, including the steps:
[0034] S1. Add 4-methoxy-N-(4-methoxyphenyl)-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)aniline and 3,8-dibromo-1,10-phenanthroline to a first solvent, add Pd(PPh)4 and K2CO3, heat and reflux under an inert atmosphere, and obtain ligand TP after the reaction;
[0035] S2. Add zinc acetate and the ligand TP to a second solvent, heat and reflux under an inert atmosphere to obtain a crude solid, and purify the crude solid to obtain the zinc complex.
[0036] In one embodiment, the molar ratio of 4-methoxy-N-(4-methoxyphenyl)-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)aniline to 3,8-dibromo-1,10-phenanthroline is 2.2:1.
[0037] In one embodiment, the first solvent includes tetrahydrofuran and water, and the volume ratio of tetrahydrofuran to water is 1:1.
[0038] In one embodiment, the molar ratio of zinc acetate to the ligand TP is 1:1.
[0039] In one embodiment, the second solvent includes toluene and methanol, and the volume ratio of toluene to methanol is 1:1.
[0040] In one embodiment, in step S1, the temperature of the heating reflux is 75 °C.
[0041] In one embodiment, in step S2, the temperature of the heating reflux is 90 °C.
[0042] The present invention also provides an application of the zinc complex in the preparation of a drug for sonodynamic therapy of tumors.
[0043] As a sonosensitizer, this zinc complex has the advantages of good biosafety, clear structure, and easy metabolism. In the absence of ultrasound, it cannot generate singlet oxygen 1 O2, and has extremely low killing ability to cells, thus having good biosafety. When activated by acoustic energy, this zinc complex can generate a large amount of singlet oxygen 1 O2 at the target site, thereby disrupting the intracellular redox balance and leading to the death of cancer cells. Since ultrasound can easily penetrate through several centimeters of tissue, tumors located deep inside the tissue can be treated. This zinc complex exhibits excellent anti-tumor effects and is expected to be used in the sonodynamic therapy of tumors.
[0044] The following specific examples are used to further illustrate the solution of the present invention.
[0045] Example 1
[0046] The synthesis route of the zinc complex Zn-TP is as follows:
[0047]
[0048]
[0049] The specific steps for preparing the zinc complex Zn-TP are as follows: Take a 50 mL reaction tube, add 4-methoxy-N-(4-methoxyphenyl)-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxolane-2-yl)phenyl)aniline (2.2 equiv.), 3,8-dibromo-1,10-phenanthroline (1 equiv.), potassium carbonate (11 equiv.), Pd(PPh)4 (0.22 equiv.) into a 6 mL mixed solvent with a volume ratio of tetrahydrofuran to deionized water of 1:1. Heat under reflux and stir overnight under nitrogen protection. After evaporating the solvent, use dichloromethane and methanol as eluents to purify the crude product by silica gel column chromatography to obtain the ligand TP with a yield of 55%. As Figure 1 shown, the 1H NMR spectrum of the product is: 1 H NMR(500MHz,Chloroform-d)δ8.37(s,2H),7.87(d,J=3.7Hz,2H),7.63(dd,J=5.3,3.3Hz,4H),7.28(d,J=4.2Hz,2H),7.20–7.15(m,8H),7.12–7.07(m,4H),6.95–6.88(m,8H),3.84(d,J=4.1Hz,12H).
[0050] Zinc acetate (CH3COO)2Zn (1 equiv.) and ligand TP (1 equiv.) were added to a solvent mixture of 5 mL of toluene and 5 mL of methanol. The mixture was heated under N2 atmosphere at 90 °C for 9 hours, cooled to room temperature, and the solvent was removed by rotary evaporation to obtain a solid with a yield of 70%. The obtained solid was dissolved in dichloromethane, and the crude product was purified by silica gel column chromatography using dichloromethane and methanol as eluents to obtain the product, i.e., zinc complex Zn-TP. As Figure 2 shown, the 1H NMR spectrum of the product is as follows: 1 H NMR (500 MHz, Chloroform-d) δ 9.42 (s, 2H), 8.51 (s, 1H), 7.95 (s, 1H), 7.56 (d, J = 8.4 Hz, 2H), 7.14 (d, J = 8.8 Hz, 4H), 7.05 (d, J = 8.6 Hz, 3H), 6.90 (d, J = 8.9 Hz, 4H), 3.83 (s, 5H), 2.36 (s, 1H).
[0051] Example 2
[0052] The zinc complex Zn-TP in Example 1 was dissolved in DMSO to prepare a mixed solvent with a concentration of 20 μM and a volume ratio of H2O:DMSO of 9:1. The UV-visible absorption spectrum was recorded on a UV-2500 spectrophotometer. The emission spectrum was recorded on an Edinburgh FS5 fluorescence spectrometer.
[0053] As Figure 3 shown in the UV-visible absorption spectrum of zinc complex Zn-TP, it can be seen that the absorption wavelength of zinc complex Zn-TP in aqueous solution (DMSO volume fraction is 10%) is 445 nm.
[0054] As Figure 4 shown in the fluorescence emission spectrum of zinc complex Zn-TP, it can be seen that the emission wavelength of zinc complex Zn-TP in aqueous solution (DMSO volume fraction is 10%) is about 600 nm.
[0055] Example 3
[0056] Using 9,10-diphenylanthracene (DPA) as a probe to detect singlet oxygen 1 O2, the ability of zinc complex Zn-TP to generate singlet oxygen under ultrasonic conditions was detected. DPA can selectively react with 1 O2, resulting in a decrease in its UV absorption value.
[0057] A mixed solution of 20 μM zinc complex Zn-TP and 10 μg / ml DPA was placed at 1.0 MHz, 3 W / cm -2, Under ultrasonic irradiation with a duty cycle of 10%, the ultraviolet absorption of DPA was measured every 10 min using an ultraviolet-visible spectrophotometer.
[0058] As Figure 5 shown, under light irradiation, the ultraviolet absorption value of DPA gradually decreased, and after 60 minutes, the absorption value almost dropped to zero, indicating that the zinc complex Zn-TP has good ability to release singlet oxygen under light conditions.
[0059] Example 4
[0060] Cytotoxicity of zinc complex Zn-TP against mouse breast cancer cells
[0061] 4T1 cells were seeded in 96-well plates and cultured for 24 hours. After the cell morphology recovered, the cells were incubated with drug-containing media with different concentrations of Zn-TP (0 μM, 1.56 μM, 3.13 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM, 100 μM) for 4 hours. The drug-containing media was aspirated, and fresh media was added. The cells were treated for 20 minutes in a dark environment and under an ultrasonic environment of 1.0 MHz, 3 W cm -2 , 10% duty cycle. Then the cells were placed in an incubator for incubation. After 44 hours, 25 μL of PBS solution containing thiazolyl blue (MTT) (5 mg mL -1 ) was added to each well, and incubation was continued for 4 hours. The culture medium was removed, and then 150 μL of DMSO was added to each well. The 96-well plate was placed on a shaker and shaken for 15 minutes. Finally, the change in optical density (OD) at 490 nm was read using an enzyme-linked immunosorbent assay reader to obtain the survival of the cells.
[0062] As Figure 6 shown, under dark conditions, the zinc complex Zn-TP has no obvious toxicity to 4T1 cells, with an IC50 > 100 μM. Under ultrasonic irradiation, the cell viability of 4T1 cells decreased significantly. The zinc complex Zn-TP showed completely different cytotoxicity under ultrasonic irradiation compared to dark conditions, with an IC50 reaching 2.45 μM, indicating that the zinc complex Zn-TP has excellent anti-tumor activity under ultrasonic irradiation.
[0063] It should be understood that the application of the present invention is not limited to the above examples. For those of ordinary skill in the art, improvements or transformations can be made according to the above description. All such improvements and transformations should fall within the protection scope of the appended claims of the present invention.
Claims
1. A zinc complex, characterized in that, The chemical structural formula of the zinc complex is as follows:
2. A method for preparing the zinc complex according to claim 1, characterized in that, It includes the steps: S1. Add 4-methoxy-N-(4-methoxyphenyl)-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxolan-2-yl)phenyl)aniline and 3,8-dibromo-1,10-phenanthroline into the first solvent, add Pd(PPh)4 and K2CO3, heat under reflux in an inert atmosphere, and obtain the ligand TP after the reaction; S2. Add zinc acetate and the ligand TP into the second solvent, heat under reflux in an inert atmosphere to obtain a crude solid, and purify the crude solid to obtain the zinc complex.
3. The preparation method of the zinc complex according to claim 2, wherein The molar ratio of the 4-methoxy-N-(4-methoxyphenyl)-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxolan-2-yl)phenyl)aniline to the 3,8-dibromo-1,10-phenanthroline is 2.2:
1.
4. The preparation method of the zinc complex according to claim 2, characterized in that, The first solvent includes tetrahydrofuran and water, and the volume ratio of the tetrahydrofuran to the water is 1:
1.
5. The preparation method of the zinc complex according to claim 2, wherein In step S2, the molar ratio of the zinc acetate to the ligand TP is 1:
1.
6. The preparation method of the zinc complex according to claim 2, wherein, The second solvent includes toluene and methanol, and the volume ratio of the toluene to the methanol is 1:
1.
7. The preparation method of the zinc complex according to claim 2, wherein In step S1, the temperature of the heating under reflux is 75 °C; in step S2, the temperature of the heating under reflux is 90 °C.
8. Use of the zinc complex according to claim 1 in the preparation of a drug for sonodynamic therapy of tumors.