CN112107597A - Copper tungstate nanodot medicine, preparation method and application in mouse tumor model - Google Patents
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Abstract
Discloses a copper tungstate nanodot drug, a preparation method thereof and application thereof in a mouse tumor model, and the copper tungstate nanodot drug can simultaneously have the synergistic effect of chemodynamics and photodynamic therapy. The method comprises the following steps: (1) adding copper salt and tungstate with the same mole into water, and stirring and dispersing uniformly; (2) adding a certain amount of sodium citrate, fully stirring and uniformly mixing; (3) transferring the mixture into a hydrothermal reaction tank for hydrothermal treatment; (4) cooling the reaction liquid; (5) taking out, dialyzing, purifying and freeze-drying to obtain the copper tungstate nanodot drug; wherein the copper salt is one or more of copper chloride, copper sulfate, copper nitrate and copper acetate; the tungstate is one or more of sodium tungstate, potassium tungstate and ammonium tungstate.
Description
Technical Field
The invention relates to the technical field of nano materials and biology, in particular to a copper tungstate nanodot drug, a preparation method thereof and application of the copper tungstate nanodot drug in a mouse tumor model.
Background
The incidence and mortality of cancer is currently on an increasing trend each year. At present, the clinical treatment of cancer mainly comprises surgical treatment and chemotherapeutic drug treatment, but the two means have great side effects. Aiming at the characteristics of cancer, the development of novel cancer treatment means is a hot direction of related research. With the development of nanotechnology, a kinetic treatment method is a promising alternative. The dynamic therapy mainly realizes the action on genetic materials in tumor cells based on active oxygen generated by chemical means or photochemical means, and achieves the purposes of cell ablation and tissue necrosis. Meanwhile, the used medicine is a catalyst, so that the property is stable, the structure cannot be changed greatly after active oxygen is generated, and extra large side effects cannot be generated after tissues are necrotized and absorbed by the body. Therefore, the method is a treatment scheme with a better application prospect.
The design of kinetic therapeutic drugs is an important aspect of kinetic therapy. Currently, there are two main kinetic drug designs, one is a chemical kinetic drug which catalyzes hydrogen peroxide existing in tumor tissue to generate active oxygen based on the catalytic properties of transition metal elements in the drug, and the other is a photodynamic drug which specifically absorbs irradiated light and effectively converts light energy into chemical energy, and hydrogen peroxide or dissolved oxygen adsorbed on or around the drug is converted into active oxygen. The chemokinetic drugs have a better killing effect on the interior of tumor tissues, while the photodynamic drugs have a better effect on cells on the surface of the tumor due to the penetration depth of light. If a nano-drug can be designed, the chemical kinetics effect and the photodynamic effect can be generated simultaneously, and the synergistic treatment effect of the two dynamics is realized, the treatment of the tumor tissue can be more effectively realized. However, few reports of drugs having both chemo-kinetic and photodynamic therapy synergistic effects are reported, which is a hot research direction for nano-drugs for cancer therapy.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide a copper tungstate nanodot drug and a preparation method thereof, which can simultaneously have the synergistic effect of chemo-kinetic and photodynamic therapy.
The technical scheme of the invention is as follows: the preparation method of the copper tungstate nanodot medicine comprises the following steps:
(1) adding copper salt and tungstate with the same mole into water, and stirring and dispersing uniformly;
(2) adding a certain amount of sodium citrate, fully stirring and uniformly mixing;
(3) transferring the mixture into a hydrothermal reaction tank for hydrothermal treatment;
(4) cooling the reaction liquid;
(5) taking out, dialyzing, purifying and freeze-drying to obtain the copper tungstate nanodot drug;
wherein the copper salt is one or more of copper chloride, copper sulfate, copper nitrate and copper acetate; the tungstate is one or more of sodium tungstate, potassium tungstate and ammonium tungstate.
A copper tungstate nanodot drug produced by the method of claims 1-8; the size of the copper tungstate nano-dot product is 2-10 nanometers.
The copper tungstate nanodot drug can dissociate copper ions under the condition of low pH value of a simulated tumor microenvironment to catalyze hydrogen peroxide to generate active oxygen, and has the application prospect of chemical kinetics; under the irradiation of exciting light with wavelength of 808 nm, hydroxyl free radicals and superoxide free radicals are generated in aqueous solution, so that the method has a photodynamic application prospect; therefore, the compound can simultaneously have the synergistic effect of chemo-kinetic and photodynamic therapy.
Also provides the application of the copper tungstate nanodot drug in a mouse tumor model.
Drawings
Fig. 1 is a Transmission Electron Microscope (TEM) image of the nanodots prepared in example 1.
Fig. 2 is a TEM image of the nanodots prepared in example 2.
Fig. 3 is a TEM image of the nanodots prepared in example 3.
Fig. 4 is data of kinetic measurements of the nanodots prepared in example 1.
FIG. 5 shows the results of the kinetic co-therapy study of nanodots prepared in example 1 in a mouse tumor model.
Fig. 6 is a flow chart of a method for preparing a copper tungstate nanodot drug according to the present invention.
Detailed Description
As shown in fig. 6, the preparation method of the copper tungstate nanodot drug comprises the following steps:
(1) adding copper salt and tungstate with the same mole into water, and stirring and dispersing uniformly;
(2) adding a certain amount of sodium citrate, fully stirring and uniformly mixing;
(3) transferring the mixture into a hydrothermal reaction tank for hydrothermal treatment;
(4) cooling the reaction liquid;
(5) taking out, dialyzing, purifying and freeze-drying to obtain the copper tungstate nanodot drug;
wherein the copper salt is one or more of copper chloride, copper sulfate, copper nitrate and copper acetate; the tungstate is one or more of sodium tungstate, potassium tungstate and ammonium tungstate.
The copper tungstate nanodot drug can dissociate copper ions under the condition of low pH value of a simulated tumor microenvironment to catalyze hydrogen peroxide to generate active oxygen, and has the application prospect of chemical kinetics; under the irradiation of exciting light with wavelength of 808 nm, hydroxyl free radicals and superoxide free radicals are generated in aqueous solution, so that the method has a photodynamic application prospect; therefore, the compound can simultaneously have the synergistic effect of chemo-kinetic and photodynamic therapy.
Preferably, in the step (1), the concentration of the copper element and the tungsten element in the reaction solution is 10 to 500 millimoles per liter.
More preferably, in the step (1), the concentration of the copper element and the tungsten element in the reaction solution is 50 to 200 millimoles per liter.
Preferably, in the step (2), the molar ratio of the initial addition amount of the sodium citrate to the use amount of the copper element in the reaction solution is 1: 0.2-1.
More preferably, in the step (2), the molar ratio of the initial addition amount of the sodium citrate to the use amount of the copper element in the reaction solution is 1: 0.5-0.8.
Preferably, in the step (3), the hydrothermal treatment is carried out in a constant temperature oven at 150-200 ℃ for 2-24 hours.
More preferably, in the step (3), the hydrothermal treatment is carried out at 190 ℃ for 6-12 hours under the condition of 170 ℃, and the reaction solution is naturally cooled to room temperature.
Preferably, in the step (5), the dialysis conditions are that a dialysis bag with molecular weight cut-off of 5000-.
More preferably, in the step (5), the dialysis conditions are 10000-.
Also provided are copper tungstate nanodot drugs, prepared by the method of claims 1-9; the size of the copper tungstate nano-dot product is 2-10 nanometers.
Also provides the application of the copper tungstate nanodot drug in a mouse tumor model.
The invention has the following beneficial effects:
1. the invention provides a preparation scheme of copper tungstate nanodots, the obtained copper tungstate nanodots can dissociate copper ions under the condition of low pH value of a simulated tumor microenvironment to catalyze hydrogen peroxide to generate active oxygen, and the preparation scheme has an application prospect of chemical kinetics.
2. The copper tungstate nanodots obtained by the method generate hydroxyl free radicals and superoxide free radicals in an aqueous solution under the irradiation of exciting light with the wavelength of 808 nanometers, and have a photodynamic application prospect.
3. The copper tungstate nanodot drug obtained by the invention is tested on a mouse tumor model, and the test result shows that the nanodot drug can generate the synergistic treatment effect of photodynamic and chemical kinetics.
The following describes in detail embodiments of the method of the present invention with reference to the accompanying drawings.
Example 1
The preparation method of the copper tungstate nanodot medicine comprises the following steps:
(1) preparation of copper tungstate reaction solution
Mixing 10ml of 50 millimole/L copper nitrate aqueous solution and 5ml of 100 millimole/L sodium tungstate aqueous solution, uniformly mixing, adding 10ml of 100 millimole/L sodium citrate aqueous solution, and continuously stirring and reacting for 1 hour.
(2) Transferring the reaction solution prepared in the step (1) into a hydrothermal reaction kettle with the volume of 50mL, and reacting for 20h at 150 ℃; and after naturally cooling to normal temperature, taking out the reaction solution, changing water for three times by using a dialysis bag with the molecular weight of 8000, dialyzing for 24 hours, and freeze-drying the purified solution to obtain the copper tungstate nanodot product.
The final product was observed with a transmission electron microscope (abbreviated as TEM, FEI Corp., USA, model Tecnai G2F 30, the same as in examples 2-4 below), and the results are shown in FIG. 1. The resulting nanodots are shown in fig. 1 to have an average particle size of 7.3 nm.
EXAMPLE 2
(1) Preparation of copper tungstate reaction solution
8ml of 500 millimole/L copper chloride aqueous solution and 8ml of 500 millimole/L potassium tungstate aqueous solution are mixed, 10ml of 2 millimole/L sodium citrate aqueous solution is added after uniform mixing, and the stirring reaction is continued for 1 hour.
(2) Transferring the reaction solution prepared in the step (1) into a hydrothermal reaction kettle with the volume of 50mL, and reacting for 2h at 200 ℃; and after naturally cooling to normal temperature, taking out the reaction liquid, changing water for three times by using a dialysis bag with the molecular weight cut-off of 20000, dialyzing for 7 hours, and freeze-drying the purified liquid to obtain the copper tungstate nanodot product.
The TEM observation of the final product obtained in example 2 is shown in FIG. 2. The resulting nanodots are shown in fig. 2 to have an average particle size of 4.2 nm.
Example 3
(1) Preparation of copper tungstate reaction solution
5ml of copper acetate aqueous solution of 100 millimoles per liter and 5ml of ammonium tungstate aqueous solution of 100 millimoles per liter are mixed, 10ml of sodium citrate aqueous solution of 100 millimoles per liter is added after uniform mixing, and stirring reaction is continued for 1 hour.
(2) Transferring the reaction solution prepared in the step (1) into a hydrothermal reaction kettle with the volume of 50mL, and reacting for 12h at 180 ℃; and naturally cooling to normal temperature, taking out the reaction solution, changing water for three times by using a dialysis bag with the molecular weight of 15000 cut-off, dialyzing for 15 hours, and freeze-drying the purified solution to obtain the copper tungstate nanodot product.
The final product obtained in this example was observed and examined as follows:
(1) observation by a transmission electron microscope:
the final product was observed by TEM and the results are shown in fig. 3. The nanodots obtained in example 3 have an average particle size of 8.4 nm as shown in fig. 3.
Example 4
Example 1 Electron spin resonance Spectroscopy testing of nanodot generated active oxygen
The nanodots obtained in example 1 were subjected to chemical kinetics and photodynamically generated active oxygen measurements using electron paramagnetic resonance spectroscopy (abbreviated as ESR, model A300-10/12, Bruker, Germany).
The results are shown in FIG. 4. In FIG. 4, the upper left panel shows the case of generating superoxide radicals by chemical kinetics, the upper right panel shows the case of generating hydroxyl radicals by chemical kinetics, the lower left panel shows the case of generating superoxide radicals by photodynamic, and the lower right panel shows the case of generating hydroxyl radicals by photodynamic. As can be seen from fig. 4, the prepared copper tungstate nanodots can effectively generate chemical kinetic effects and photodynamic effects.
Example 5:
therapeutic applications of mouse tumor models:
(1) the copper tungstate nanodots obtained in example 1 were added to a biological phosphate buffer solution (PBS for short) to prepare a PBS solution having a copper element concentration of 20 μ g per ml.
(2) And (3) constructing a leg subcutaneous tumor model mouse with the tumor volume of about 100 cubic centimeters. Injecting the PBS solution of the nanodots obtained in the step 1 into the tumor, after the solution is well dispersed after half an hour, generating near infrared light with the wavelength of 808 nanometers at a fixed point of the tumor part, wherein the irradiation intensity is 1 watt per square centimeter, and the irradiation is carried out for 5 minutes. After two weeks of subsequent culture, tumor tissues were removed to observe changes in tumor volume in the mice.
A contrast photograph of the subcutaneous tumor taken after dissection is shown in fig. 5, and the three group tumor tissue photographs in the uppermost row of fig. 5 are blank control groups; the second group of tumor tissue photographs from top to bottom are the group irradiated with only 808 nm wavelength near-infrared light, the third group is the group injected with the nanodot material obtained in example 1, there is no group irradiated with light, and the bottom group is the group injected with the nanodot material obtained in example 1 and irradiated with 808 nm wavelength near-infrared light. As can be seen from fig. 5, the nanodot material obtained in example 1 was injected and irradiated with near infrared light, which effectively inhibited the growth of mouse tumor tissue.
Through a plurality of experiments, the size of the final copper tungstate nanodot product obtained by the preparation method is 2-10 nanometers.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in any way, and all simple modifications, equivalent variations and modifications made to the above embodiment according to the technical spirit of the present invention still belong to the protection scope of the technical solution of the present invention.
Claims (10)
1. The preparation method of the copper tungstate nanodot medicine is characterized by comprising the following steps: which comprises the following steps:
(1) adding copper salt and tungstate with the same mole into water, and stirring and dispersing uniformly;
(2) adding a certain amount of sodium citrate, fully stirring and uniformly mixing;
(3) transferring the mixture into a hydrothermal reaction tank for hydrothermal treatment;
(4) cooling the reaction liquid;
(5) taking out, dialyzing, purifying and freeze-drying to obtain the copper tungstate nanodot drug;
wherein the copper salt is one or more of copper chloride, copper sulfate, copper nitrate and copper acetate; the tungstate is one or more of sodium tungstate, potassium tungstate and ammonium tungstate.
2. The method for preparing a copper tungstate nanodot drug as claimed in claim 1, wherein: in the step (1), the concentration of copper element and tungsten element in the reaction solution is 10-500 millimoles per liter.
3. The method for preparing a copper tungstate nanodot drug as claimed in claim 2, wherein: in the step (1), the concentration of copper element and tungsten element in the reaction solution is 50-200 millimoles per liter.
4. The method for preparing a copper tungstate nanodot drug as claimed in claim 3, wherein: in the step (2), the molar ratio of the initial addition amount of the sodium citrate to the use amount of the copper element in the reaction solution is 1: 0.2-1.
5. The method for preparing a copper tungstate nanodot drug as claimed in claim 4, wherein: in the step (2), the molar ratio of the initial addition amount of the sodium citrate to the use amount of the copper element in the reaction solution is 1: 0.5-0.8.
6. The method for preparing a copper tungstate nanodot drug as claimed in claim 5, wherein: in the step (3), the hydrothermal treatment is carried out in a constant temperature oven with the temperature of 150 ℃ and 200 ℃ for 2-24 hours.
7. The method for preparing a copper tungstate nanodot drug as claimed in claim 6, wherein: in the step (3), the hydrothermal treatment is carried out at 190 ℃ for 6-12 hours under the condition of 170 ℃, and the reaction is naturally cooled to room temperature.
8. The method for preparing a copper tungstate nanodot drug as claimed in claim 7, wherein: in the step (5), the dialysis condition is to select a dialysis bag with molecular weight cut-off of 5000-.
9. The copper tungstate nanodot medicine is characterized in that: prepared by the process of claims 1-8; the size of the copper tungstate nano-dot product is 2-10 nanometers.
10. Use of a copper tungstate nanodot drug according to claim 9 in a mouse tumor model.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015195889A2 (en) * | 2014-06-18 | 2015-12-23 | University Of Georgia Research Foundation, Inc. | Induced photodynamic therapy using nanoparticle scintillators as transducers |
| CN108069458A (en) * | 2017-12-26 | 2018-05-25 | 南京鼓楼医院 | A kind of ultra micro nano-level sphere bismuth tungstate crystal grain and preparation method and application |
| CN111450081A (en) * | 2020-04-10 | 2020-07-28 | 大连理工大学 | Protein nanoparticle for enhanced dynamic therapy, preparation method and application thereof |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015195889A2 (en) * | 2014-06-18 | 2015-12-23 | University Of Georgia Research Foundation, Inc. | Induced photodynamic therapy using nanoparticle scintillators as transducers |
| CN108069458A (en) * | 2017-12-26 | 2018-05-25 | 南京鼓楼医院 | A kind of ultra micro nano-level sphere bismuth tungstate crystal grain and preparation method and application |
| CN111450081A (en) * | 2020-04-10 | 2020-07-28 | 大连理工大学 | Protein nanoparticle for enhanced dynamic therapy, preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| WEN ET AL.: "Tuning the NIR photoabsorption of CuWO 4-x nanodots with oxygen vacancies for CT imaging guided photothermal therapy of tumors", 《BIOMATER SCI》 * |
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Application publication date: 20201222 |