Preparation method of multi-component nano-drug based on ruthenium complex and application of multi-component nano-drug in tumor resistance
The preparation of multi-component nanomedicines of ruthenium complexes by solvent-antisolvent method has solved the problems of poor water solubility and low bioavailability, realizing highly efficient anti-tumor therapy of ruthenium complexes, overcoming drug resistance, and providing a safe and efficient tumor treatment strategy.
Patent Information
- Application Number
- CN202511232123.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-01
- Publication Date
- 2025-11-21
AI Technical Summary
Ruthenium complexes have limitations in antitumor therapy due to their poor water solubility, low bioavailability, and the tendency to develop drug resistance when used as a single agent, which restricts their clinical application.
A solvent-antisolvent method was used to prepare multi-component nanomedicines based on ruthenium complexes. By self-assembling ruthenium complexes, oxidative stress amplifiers and DNA repair inhibitors, carrier-free fully active nanomedicines were formed. A nanodelivery system was used to improve bioavailability and achieve synergistic therapy.
It improves the water solubility and bioavailability of ruthenium complexes, enhances antitumor effects, overcomes drug resistance, and provides a safe and efficient tumor treatment strategy.
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Abstract
Description
TECHNICAL FIELD
[0001] The application relates to a preparation method of a multi-component nanodrug based on a ruthenium complex and application thereof in anti-tumor, and belongs to the field of biological medicines. BACKGROUND
[0002] Platinum-based chemotherapy drugs are often used as first-line drugs in tumor treatment, however, due to their significant toxic side effects and the fact that long-term use can easily make tumor cells resistant, the clinical application of the drugs is greatly limited. In comparison, ruthenium complexes, which also belong to transition metals, exhibit unique advantages, many of which not only have lower toxicity, but also show high selectivity to tumor cells, and are expected to break through the application bottleneck of existing platinum drugs. In particular, ruthenium polypyridyl complexes, with their unique photo-physical and photo-chemical properties and easy-to-modify molecular structure, not only have potential application prospects in traditional chemotherapy, but also have attracted widespread attention in the application of photoactivated anti-tumor. From the mechanism of action, such compounds have multiple potential anti-tumor pathways when applied to photoactivated anti-tumor. One is to achieve oxidative damage to tumor cells by generating reactive oxygen species (ROS) under irradiation of specific wavelength light, oxidizing and breaking down biological macromolecules such as DNA, i.e. to achieve photodynamic anti-tumor; the other is to generate hydrolysis products by photolabile ligand dissociation, which can cross-link with DNA bases or other biological macromolecules, and then kill tumor cells.
[0003] Although ruthenium complexes have excellent anti-tumor activity, their poor water solubility, low bioavailability and single drug use easily leading to drug resistance limit their clinical application.
[0004] Synergistic drug strategy is often used as a means to overcome the problems of single drug use. At the same time, nanodrug delivery systems can effectively improve the bioavailability of drugs by improving the water solubility of drugs and achieving targeted transport. The integration of nanodrug delivery systems and synergistic drug strategies not only can solve the problems of low bioavailability and drug resistance of ruthenium complexes, but also can bring synergistic effects, and can provide new ideas for developing safe and efficient tumor treatment strategies. SUMMARY
[0005] The main purpose of the present application is to solve the problems of poor water solubility, low bioavailability and single drug use easily leading to drug resistance of ruthenium complexes, and to provide a preparation method of a multi-component nanodrug based on a ruthenium complex, and to improve the anti-tumor effect thereof by synergistic drug strategy and nanotechnology.
[0006] The technical scheme adopted to achieve the purpose of the present application is:
[0007] One of the purposes of the present application is to provide a preparation method of a multi-component nanodrug based on a ruthenium complex, which is a solvent-anti-solvent method, and the ruthenium complex is self-assembled with a plurality of active drugs by means of intermolecular weak interaction to construct a nanodrug of a full-activity multi-drug combination without a carrier, and the preparation method comprises the following steps:
[0008] (1) Dissolving a poorly soluble drug in an appropriate amount of organic solvent, wherein the poorly soluble drug comprises a ruthenium complex, an oxidative stress amplifier and a DNA repair inhibitor.
[0009] (2) Mixing the ruthenium complex solution, the oxidative stress amplifier and the DNA repair inhibitor in step (1) according to a certain molar ratio for a certain time, dropping the mixed solution into an appropriate amount of poor solvent ultrapure water, and mixing and stirring in the dark for a certain time to obtain a multi-component nanodrug composed of the ruthenium complex, the oxidative stress amplifier and the DNA repair inhibitor.
[0010] (3) Dialyzing the multi-component nanodrug formed in step (2) for 6h to remove the molecular state drug components to obtain a multi-component nanodrug based on the ruthenium complex, named RuOCANps, and the nanodrug is stored in a solution state at 4℃, wherein the molecular weight cut-off of the dialysis bag used is 1500.
[0011] Preferably, the organic solvent in step (1) is methanol.
[0012] Preferably, the ruthenium complex in step (1) is a ruthenium polypyridine complex [Ru(dip)2(tpy-Py)2](PF6)2 (dip = 4,7-diphenyl-1,10-phenanthroline, tpy-Py = pyrene-modified 2,2':6',2"-terpyridine ligand), which generates ROS and undergoes ligand dissociation under light; the oxidative stress amplifier is cinnamaldehyde, which amplifies the generation of ROS in cooperation with the ruthenium complex; and the DNA repair inhibitor is olaparib, which inhibits DNA repair.
[0013] Preferably, the mixing time of the ruthenium complex solution, cinnamaldehyde and olaparib in step (2) is 2h-10h, more preferably 10h.
[0014] Preferably, the concentration of the ruthenium complex in the poorly soluble drug in step (1) is 1mg / mL, the concentration of cinnamaldehyde is 2mg / mL, and the concentration of olaparib is 1mg / mL.
[0015] Preferably, the molar ratio of the ruthenium complex, cinnamaldehyde and olaparib in step (2) is 1:2:1-1:2:3, more preferably 1:2:1.
[0016] Preferably, the volume of the poor solvent ultrapure water in step (2) is 1-3 mL, more preferably 2 mL.
[0017] Preferably, the mixing and stirring speed in step (2) is 950 rpm / min.
[0018] Preferably, in step (2), after the mixed solution is dropped into the ultrapure water, the mixing and stirring time is 4-12 h, more preferably 10 h.
[0019] With the above technical solutions, the application has the following technical effects:
[0020] The application provides a multi-component nanodrug based on a ruthenium complex, the multi-component nanodrug RuOCANps is a fully active nanodrug, has no inert carrier component, is uniform and stable in preparation, can be stored for a long time, has good biological safety, can improve the problems of poor water solubility and low bioavailability of the ruthenium complex and the problem that single drug use is easy to cause drug resistance, and further enhances the anti-tumor effect by means of multi-drug synergistic effect, thereby expanding a new path for the treatment of malignant tumors. BRIEF DESCRIPTION OF DRAWINGS
[0021] Figure 1 is a structural formula of the ruthenium polypyridine complex [Ru(dip)2(tpy-Py)2](PF6)2, cinnamyl aldehyde and olaparib described in the embodiment of the application.
[0022] Figure 2 is a transmission electron microscope image of the multi-component nanodrug RuOCANps based on a ruthenium complex obtained in Example 1 of the application.
[0023] Figure 3 is a particle size distribution diagram of the multi-component nanodrug RuOCANps based on a ruthenium complex obtained in Example 1 of the application.
[0024] Figure 4 (a) is a stability characterization diagram of the multi-component nanodrug RuOCANps based on a ruthenium complex solution prepared in Example 1 within 5 days; Figure 4 (b) is a stability characterization diagram of the multi-component nanodrug RuOCANps based on a ruthenium complex dispersed in a culture medium containing 10% fetal bovine serum within 5 days.
[0025] Figure 5 (a) is a cytotoxicity result of the multi-component nanodrug RuOCANps based on a ruthenium complex and a reference drug on human lung cancer cells (A549) in dark conditions, obtained in Example 1 of the application. Figure 5 (b) is a cytotoxicity result of the multi-component nanodrug RuOCANps based on a ruthenium complex and a reference drug on A549 cells under light conditions.
[0026] Figure 6 The figure of the generation of reactive oxygen species (ROS) in A549 cells by the multi-component nanomedicine RuOCANps based on a ruthenium complex obtained in Embodiment 1 of the present application and the reference drug.
[0027] Figure 7 The figure of the results of the live and dead cell staining experiment of A549 cells by the multi-component nanomedicine RuOCANps based on a ruthenium complex obtained in Embodiment 1 of the present application and the reference drug under dark and light (λ = 470 nm) conditions. DETAILED DESCRIPTION
[0028] The technical solutions of the present application will be described in detail below with reference to specific embodiments and the accompanying drawings, so as to facilitate the understanding of those skilled in the art, but not in any way limit the present application.
[0029] Embodiment 1
[0030] A multi-component nanomedicine based on a ruthenium complex, the specific steps of the preparation method are as follows:
[0031] The ruthenium polypyridyl complex [Ru(dip)2(tpy-Py)2](PF6)2, cinnamaldehyde and olaparib were respectively dissolved with a good solvent methanol to prepare three solutions with concentrations of 1 mg / mL, 2 mg / mL and 1 mg / mL, then 100 μL of the ruthenium complex solution, 8.9 μL of the cinnamaldehyde solution and 29.1 μL of the olaparib solution (molecular molar ratio of 1:2:1) were mixed for 10 h, and then slowly dropped into 2 mL of a poor solvent ultrapure water, and mixed and stirred under dark conditions for 10 h, and then the mixture was transferred to a dialysis bag with a molecular weight cut-off of 1500, and dialyzed in ultrapure water for 6 h, and then the liquid in the dialysis bag was collected to obtain a multi-component nanomedicine based on a ruthenium complex, and the obtained nanomaterial was stored at 4℃.
[0032] Embodiment 2
[0033] A multi-component nanomedicine based on a ruthenium complex, the specific steps of the preparation method are as follows:
[0034] Ruthenium polypyridine complex [Ru(dip)2(tpy-Py)2](PF6)2, cinnamaldehyde, and olaparib were dissolved in methanol, a good solvent, to prepare three solutions with concentrations of 1 mg / mL, 2 mg / mL, and 1 mg / mL, respectively. Then, 100 μL of the ruthenium complex solution, 8.9 μL of the cinnamaldehyde solution, and 58.2 μL of the olaparib solution (molecular molar ratio of 1:2:2) were mixed for 10 h, and then 2 mL of ultrapure water, a poor solvent, was slowly added dropwise. The mixture was shaken and stirred for 10 h in the dark. The mixture was then transferred to a dialysis bag with a molecular weight cutoff of 1500 and dialyzed in ultrapure water for 6 h. The liquid in the dialysis bag was collected to obtain a multi-component nanomedicine based on the ruthenium complex. The obtained nanomaterials were stored at 4 °C.
[0035] Example 3
[0036] A multi-component nanomedicine based on ruthenium complexes, the specific steps of which are as follows:
[0037] Ruthenium polypyridine complex [Ru(dip)2(tpy-Py)2](PF6)2, cinnamaldehyde, and olaparib were dissolved in methanol, a good solvent, to prepare three solutions with concentrations of 1 mg / mL, 2 mg / mL, and 1 mg / mL, respectively. Then, 100 μL of the ruthenium complex solution, 8.9 μL of the cinnamaldehyde solution, and 87.3 μL of the olaparib solution (molecular molar ratio of 1:2:3) were mixed for 10 h. Then, 2 mL of ultrapure water, a poor solvent, was slowly added dropwise. The mixture was shaken and stirred for 10 h in the dark. The mixture was then transferred to a dialysis bag with a molecular weight cutoff of 1500 and dialyzed in ultrapure water for 6 h. The liquid in the dialysis bag was collected to obtain a multi-component nanomedicine based on the ruthenium complex. The obtained nanomaterials were stored at 4 °C.
[0038] The ruthenium complex-based multi-component nanomedicine RuOCANps prepared in the above examples were analyzed as follows:
[0039] (1) Morphological analysis
[0040] The hydrated particle size of the multi-component nanomedicines composed of cinnamaldehyde, ruthenium complex, and olaparib in Examples 1, 2, and 3 was characterized using a Malvern particle size analyzer. Figure 2 The particle size distribution diagram for Example 1 shows that the hydrated particle size of the nanocomposite is approximately 195 nm, and it exhibits good dispersibility.
[0041] The morphology of the ruthenium complex-based multidrug nanomaterials obtained in Examples 1, 2, and 3 was characterized using transmission electron microscopy (TEM). Figure 3 This is a TEM image of Example 1. As can be seen from the image, the nanomaterial particles have a diameter of approximately 50 nm and are uniformly spherical.
[0042] (2) Stability test
[0043] The multi-drug composition nanomaterial solution prepared in Example 1 was stored at 4°C and dispersed in a culture medium containing 10% fetal bovine serum for 5 days to observe its stability. The results are as follows: Figure 4 As shown in (a), no obvious aggregation or precipitation was observed in the nanomedicine solution over the 5 days. Figure 4 As shown in (b), no significant aggregation occurred when dispersed in a culture medium containing 10% fetal bovine serum, demonstrating its good particle stability.
[0044] (3) In vitro antitumor activity
[0045] The CCK8 assay was used to evaluate the cytotoxic effect of the ruthenium complex-based multicomponent nanomedicine RuOCANps prepared in Example 1 on in vitro tumor cells, with a physical mixture of the ruthenium complex and cinnamaldehyde in the same molar ratio and each monomeric drug as references. Figure 5 As shown in (a), the multi-component nanomedicine exhibits low dark toxicity to A549 cells under dark conditions. Figure 5 As shown in (b), the survival rate of A549 cells cultured in RuOCANps under light conditions decreased with increasing concentration, and the phototoxicity was greater than that of the monomeric ruthenium complex and the synergistic effect of the ruthenium complex and cinnamaldehyde. This is because the ruthenium complex had a synergistic effect with cinnamaldehyde and olaparib in RuOCANps.
[0046] The cytotoxic effect of RuOCANps, a multi-component nanomedicine based on ruthenium complexes prepared in Example 1, on in vitro tumor cells was verified using a live-dead staining experiment. After drug treatment, the cells were stained with a biological cell viability assay reagent (Calcein AM concentration of 1 μM, PI concentration of 4 μM, live cells appearing green and dead cells appearing red), and the staining results were detected by fluorescence microscopy. The results are as follows: Figure 6 As shown, under light conditions, RuOCANps exhibited greater cytotoxicity to A549 cells than the monomeric ruthenium group and the synergistic effect of the ruthenium complex and cinnamaldehyde. These results are consistent with those of CCK8, indicating that the nanomedicine group has a better tumor-killing effect.
[0047] (4) The ability to generate reactive oxygen species (ROS) in tumor cells under light conditions.
[0048] The ability of the multi-component nanomedicine RuOCANps, based on ruthenium complexes prepared in Example 1, to generate ROS in A549 cells was observed using a fluorescence microscope using the ROS probe DCFH-DA. Figure 7As shown, compared to the blank group where no ROS green fluorescence signal was observed, the RuOCANps group showed a significant ROS signal, and the ROS green fluorescence signal of the RuOCANps group was significantly stronger than that of the monomeric ruthenium group, indicating that it has a better ability to generate ROS.
Claims
1. A method for preparing a multi-component nanomedicine based on ruthenium complexes and its application in antitumor treatment, characterized in that: The aforementioned multi-component nanomedicine based on ruthenium complexes combines ruthenium complexes and two synergistic drugs through weak intermolecular forces. The ruthenium complex is a ruthenium polypyridine complex [Ru(dip)2(tpy-Py)2](PF6)2, and the two synergistic drugs are cinnamaldehyde and the DNA repair inhibitor olaparib. The multi-component nanomedicine based on the ruthenium complex is used in the preparation of anti-lung cancer drugs.
2. A method for preparing a multi-component nanomedicine based on ruthenium complexes and its application in antitumor treatment, characterized in that... The preparation method employs a solvent-antisolvent method, and the steps of which are as follows: (1) Dissolve the ruthenium complex and synergistic drugs cinnamaldehyde and olaparib in an appropriate amount of organic solvent, and mix a certain amount of the ruthenium complex, cinnamaldehyde and olaparib for a certain period of time. (2) Slowly drop the mixture from step (1) into the antisolvent ultrapure water, and continue to shake and mix for a certain period of time under dark conditions to obtain the nanomedicine to be purified. (3) Transfer the mixture from step (2) to a dialysis bag for dialysis purification, and collect the liquid in the dialysis bag.
3. The method for preparing a multi-component nanomedicine based on a ruthenium complex according to claim 2, characterized in that: The organic solvent in step (1) is methanol, and the selected molar ratio of ruthenium complex, cinnamaldehyde and olaparib is 1:2:1 to 1:2:
3. The mixing time of the three drugs is 10h.
4. The method for preparing a multi-component nanomedicine based on a ruthenium complex according to claim 2, characterized in that: The oscillation mixing speed in step (2) is 950 rpm / min, and the mixing time of the mixture is 10 h.
5. The method for preparing a multi-component nanomedicine based on a ruthenium complex according to claim 2, characterized in that: The dialysis time in step (3) is 6 hours, and the molecular weight cutoff of the dialysis bag used is 1500.
6. The application of the multi-component nanomedicine based on ruthenium complex as described in claim 1 in antitumor activity.