CN112675150B - Preparation method of tumor-targeted drug-loaded nanoparticles based on stibene - Google Patents
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Abstract
The invention relates to the field of medicines, and discloses a preparation method of a tumor targeted drug-loaded nanoparticle based on stibene. The preparation method comprises the steps of firstly obtaining stibene nanoparticles through a liquid phase mechanical stripping method and differential centrifugation, then coating chitosan and adriamycin with positive charges on the surfaces of AMNPs with negative charges by utilizing electrostatic adsorption, and finally coating hyaluronic acid with negative charges on the surfaces of chitosan to obtain the photo-thermal/chemotherapy drug-loaded nanoparticles capable of realizing tumor targeting, drug slow release and multi-modal imaging. The drug-loaded nano particle has important development prospect in the aspects of anti-tumor photothermal photodynamic therapy, drug nano carriers and the like.
Description
Technical Field
The invention relates to the field of medicines, in particular to a preparation method of a tumor targeted drug-loaded nanoparticle based on stibene.
Background
The traditional tumor treatment method has high recurrence and great side effect, can not realize accurate treatment of malignant tumor, and has certain damage to normal tissues. With the rise of new tumor treatment modes of photothermal photodynamic therapy, the search for suitable materials has become a primary task. Since the first report of antimonene in 2015, the antimonene gradually attracts wide attention due to excellent physicochemical properties, including application prospect in cancer treatment. Although the antimonene nano material (AM) is a new photo-thermal therapeutic agent which appears recently, the direct utilization of the antimonene nano material is greatly limited by the rapid degradation of the antimonene nano material in a physiological medium, and the antimonene nano material is rarely reported in the aspect of tumor treatment; secondly, AM surface active groups are few, chemical modification difficulty is large, and the stability of products obtained by combination of Van der Waals force and hydrogen bond hydrophobic effect is poor. Therefore, the application of the stibene in cancer treatment provides a new direction for the research of antitumor nano-drugs.
Disclosure of Invention
In order to solve the technical problems, the invention provides a preparation method of a tumor targeted drug-loaded nanoparticle based on stibene. The preparation method comprises the steps of firstly obtaining stibene nanoparticles through a liquid-phase mechanical stripping method and differential centrifugation, then coating chitosan with positive charges and adriamycin on the surfaces of the stibene nanoparticles with negative charges by utilizing the electrostatic adsorption effect, and finally coating hyaluronic acid with negative charges on the surfaces of chitosan. The nano particles obtained by the method have multiple functions, can realize photoacoustic imaging, fluorescence imaging, photo-thermal/chemotherapy synergistic treatment and tumor targeting, can realize the function of slow release of the drug, increase the stability of the nano drug taking stibene as a matrix, enhance the anti-tumor effect and are expected to be actually applied to tumor treatment; meanwhile, the method does not have obvious influence on cells in subsequent experiments, does not influence the scientificity of experimental results, and is simple, safe, nontoxic and environment-friendly in preparation process operation.
The specific technical scheme of the invention is as follows: a preparation method of a tumor targeted drug-loaded nanoparticle based on stibene comprises the following steps:
(1) Preparing a chitosan solution: dissolving chitosan in acetic acid to prepare a chitosan solution with the concentration of 1-2mg/mL, and refrigerating for later use.
(2) Preparing a hyaluronic acid solution: dissolving hyaluronic acid in deionized water to prepare hyaluronic acid with concentration of 1-2mg/mL, and refrigerating for later use.
(3) Preparing an adriamycin solution: dissolving adriamycin in deionized water to prepare adriamycin solution of 1-2mg/mL, encapsulating with tinfoil, and refrigerating for later use.
(4) Preparation of AMNPs: dispersing 2-4g of antimony powder in 40-80 mLN-methyl-2-pyrrolidone, sealing the container, performing ultrasonic treatment with an ultrasonic probe in an ice bath through an ultrasonic cell crushing instrument, observing the temperature of the ice bath at intervals of 20-40min, continuing ultrasonic treatment after stirring, and performing ultrasonic treatment in an ice bath in an ultrasonic cleaner; and finally, carrying out ultrasonic treatment by using an ultrasonic cell crusher.
Centrifuging the obtained dispersion liquid at 8-10 ℃, discarding deposited blocky antimony, centrifuging again, collecting precipitate, adding 10-15mL of deionized water to obtain the stibene nano-particle dispersion liquid, and refrigerating for later use.
(5) Diluting the stibene nano-particle dispersion liquid obtained in the step (4), measuring the concentration by using an ultraviolet-visible light spectrometer, and diluting to prepare the AMNPs dispersion liquid with the concentration of 1-2 mg/mL.
(6) Chitosan and doxorubicin coated AMNPs: adding 0.5-1mL of chitosan solution and 0.5-1mL of adriamycin solution into 0.5-1mL of AMNPs dispersion liquid, stirring, centrifuging, collecting precipitate, and adding 1-2mL of deionized water to obtain AMNPs @ CS-DOX nanoparticle dispersion liquid.
(7) Coating hyaluronic acid: and adding 1-2mL of hyaluronic acid solution into the obtained nanoparticle dispersion liquid, performing ultrasonic treatment in an ultrasonic cleaner, stirring, aging overnight, centrifuging twice, collecting precipitates, and finally obtaining the tumor-targeted drug-loaded nanoparticles based on the stibene.
The preparation method comprises the steps of firstly obtaining stibene nanoparticles through a liquid-phase mechanical stripping method and differential centrifugation, then coating chitosan and adriamycin with positive charges on the surfaces of the stibene nanoparticles with negative charges by utilizing the electrostatic adsorption effect, and finally coating hyaluronic acid with negative charges on the surfaces of chitosan.
Antimony is in the same family as phosphorus and has similar morphological and functional properties. The stibene has a honeycomb structure similar to BP, can be used for loading and conveying medicaments, has excellent photo-thermal conversion efficiency, and has environmental stability obviously better than BP. In addition, the stibene nanoparticles have excellent acousto-optic performance, and show the strongest photoacoustic signal in a 2D material family, and the photoacoustic imaging performance is endowed by the characteristics; and the stibene nano-particles can be converted into Sb with specific cytotoxicity to cancer cells under the irradiation of X-rays 2 O 3 And Sb 2 O 5 ，Sb 2 O 3 The strong cytotoxicity effect is exerted by inducing the excessive generation of active oxygen and the damage of mitochondria, and the characteristic endows the stibene nanoparticles with the potential in the photodynamic treatment of tumors; the spontaneous oxidation of the stibene accelerates the release of light energy, further improves the photothermal conversion by accelerating the non-radiative transition rate, and widens the absorption range of a near infrared region, so that the stibene nanoparticles have potential in photothermal treatment of tumors.
Based on the functions of the stibene, the chitosan with good biocompatibility and biodegradability is introduced, so that the biocompatibility and the structural stability of the nano particles in the physiological environment are improved; the loaded adriamycin has a fluorescent effect; the coated hyaluronic acid further optimizes the biological characteristics of the stibene, improves the blood compatibility and enables the nano material to have the functions of tumor targeting and drug sustained release. The finally obtained nano-drug can accurately treat local tumors, has photothermal/chemotherapy synergistic treatment effect and multimode imaging property (fluorescence/photoacoustic/photothermal imaging) and provides a new direction for nano-drug research of antitumor treatment.
Preferably, in the step (4), the first centrifugation rotating speed is 2000-4000rpm, and the centrifugation time is 10-15min; the second centrifugation rotating speed is 8000-12000rpm, and the centrifugation time is 30-35min; the centrifugation temperature does not exceed 10 ℃. Preferably, in the step (4), the specification of the ultrasonic probe is phi 6, the ice-bath ultrasonic time is 8-10h, the power is 800-1200w, and the ultrasonic operation is stopped for 1s every 2-4 s; the ultrasonic cleaner is used for carrying out ice bath and ultrasonic treatment for 2-3h and has the power of 200-400w; the ice-bath ultrasonic time of the ultrasonic cell crusher is 2-3h.
Preferably, in the steps (1) to (4), the refrigerating temperature is 1-5 ℃.
Preferably, in the step (6), the stirring time is 8-12h; the centrifugal speed is 10000-12000rpm, and the centrifugal time is 3-5min.
Preferably, in the step (7), the ultrasonic time is 15-20min, the magnetic stirring time is 3-5h, and the mixture is aged overnight; the two-time centrifugation rotating speed is 10000-12000rpm, and each time of centrifugation is 3-5min.
Compared with the prior art, the invention has the beneficial effects that:
1. the invention successfully prepares the stibene nano-particles by liquid phase stripping and differential centrifugation.
2. Good biocompatibility, biological safety and physiological environment stability are important indexes for inspecting the superiority of the photothermal agent. According to the invention, the chitosan is coated outside the stibene nano-particles by utilizing the electrostatic adsorption effect, so that the biocompatibility and the structural stability of the nano-particles are improved, and the degradation of the material in a physiological environment is slowed down, namely the stability of the material in the physiological environment is improved.
3. Compared with graphene nanosheets, BP and transition metal sulfide nanosheets, the antimonene nanosheets have higher photo-thermal conversion efficiency and excellent photo-thermal performance, have photodynamic performance due to the spontaneous oxidation of the antimonene, endow the material with a photo-thermal and photodynamic synergistic treatment effect, and meanwhile, the surface of the material is loaded with adriamycin, so that the anti-tumor effect of the nano-drug is enhanced.
4. The stibene used in the invention has the function of photoacoustic imaging, and the loaded adriamycin enables the whole nano material to have the effect of fluorescence imaging, so that the material is endowed with the effects of fluorescence, photothermal and photoacoustic imaging, and the accuracy of local treatment of nano drugs is greatly improved.
5. The hyaluronic acid is coated on the outermost layer of the micelle, so that the tumor targeting capability is enhanced, the blood compatibility and the drug sustained release of the nano-drug are greatly improved, and the precise release of the chemotherapeutic drug to a tumor part is realized.
6. The invention introduces the novel 2D material stibene into the field of anti-tumor nano-drugs, and opens up a new path for the application research of the stibene in a photo-thermal agent.
Drawings
FIG. 1 is a photothermal plot of AM @ CS/DOX @ HA as prepared in example 1 of the present invention;
FIG. 2 is the photodynamic graph of AM @ CS/DOX @ HA prepared in example 1 of the present invention;
FIG. 3 is a TEM image of AM prepared in example 1 of the present invention; FIG. 4 is a graph of AM @ CS/DOX @ HA drug release profile prepared in example 1 of the present invention.
Detailed Description
The present invention will be further described with reference to the following examples.
Example 1
(1) Preparing a chitosan solution: 20mg of chitosan was weighed and dissolved in 20mL of acetic acid to prepare a 1mg/mL chitosan solution, which was stored in a refrigerator at 4 ℃.
(2) Preparing a hyaluronic acid solution: 20mg of hyaluronic acid is weighed and dissolved in 20mL of deionized water to prepare a hyaluronic acid solution with the concentration of 1mg/mL, and the hyaluronic acid solution is stored in a refrigerator at the temperature of 4 ℃.
(3) Preparing an adriamycin solution: 10mg of ICG was weighed and dissolved in 10mL of deionized water to prepare a 1mg/mL doxorubicin solution, which was then stored in a refrigerator at 4 ℃ with a foil envelope.
(4) Preparation of AMNPs: 2g of antimony powder is taken and dispersed in a clean beaker filled with 40ml of NMP at room temperature, a preservative film is sealed, a phi 6 ultrasonic probe is used for carrying out ice bath ultrasonic treatment for 8 hours under an ultrasonic cell crushing instrument, the power is 1000w, the ultrasonic treatment is stopped for 1s every 3s, a glass rod is used for stirring dispersion liquid every 30min to ensure that the dispersion liquid is uniformly dispersed, whether ice blocks are dissolved in ice bath is observed, the ice blocks are supplemented in time, then the ice bath ultrasonic treatment is carried out for 2 hours in an ultrasonic cleaner, the power is 300w, and the ultrasonic cell crushing instrument is used for carrying out ultrasonic treatment for 2 hours; and finally, centrifuging: centrifuging the obtained dispersion liquid at the rotating speed of 3000rpm for 10min at the temperature of 8 ℃, discarding deposited blocky antimony, retaining supernatant, centrifuging the supernatant at the rotating speed of 8000rpm for 45min, collecting precipitate, adding 10ml of deionized water to obtain stibene nanoparticle dispersion liquid, and storing the stibene nanoparticle dispersion liquid at the temperature of 4 ℃.
After being diluted properly, the AMNPs dispersion liquid is measured to have the absorbance under the wavelength of 808nm by an ultraviolet-visible light analyzer, the concentration of antimony in the AMNPs dispersion liquid prepared by the method is calculated to be 7mg/mL according to an antimony standard curve, and the AMNPs dispersion liquid with the concentration of 1mg/mL is obtained after being diluted by 7 times and is used for subsequent experiments.
(5) CS and DOX coated AMNPs: adding 1mL of chitosan solution and 1mL of DOX into 1mL of AMNPs dispersion liquid, magnetically stirring for 8h, centrifuging at 10000rpm for 3min, collecting precipitate, adding 1mL of deionized water ultrasonic cleaner, and ultrasonically dispersing for 10min to obtain AMNPs @ CS-DOX nanoparticle dispersion liquid.
(6) Coated Hyaluronic Acid (HA): adding 1mL of hyaluronic acid solution into the obtained nanoparticle dispersion liquid, performing ultrasonic treatment for 15min by using an ultrasonic cleaner, performing magnetic stirring for 3h, aging overnight, centrifuging at 10000rpm for 3min, collecting precipitate, adding 1mL of deionized water, performing ultrasonic dispersion for 10min, centrifuging again at the same centrifugal parameters, collecting precipitate, adding 1mL of deionized water, finally obtaining 1mg/mL of drug-loaded nanoparticle AM @ CS/DOX @ HA dispersion liquid, and refrigerating in a refrigerator at 4 ℃.
FIG. 1 is a photothermal graph of AM @ CS/DOX @ HA prepared in example 1 of the present invention at 808nm,1.5Wcm -2 The temperature can reach nearly 90 ℃ after the laser with power density irradiates for 200s, and the photo-thermal performance is proved to be better; FIG. 2 is a graph of the photodynamic AM @ CS/DOX @ HA profile prepared in example 1 of the present invention, which shows that the active oxygen generation efficiency is high, and the AM @ CS/DOX @ HA has good photodynamic performance; fig. 3 is a TEM image of AM prepared in example 1 of the present invention, illustrating that the prepared AM has a sheet structure; FIG. 4 is a graph showing the release profile of AM @ CS/DOX @ HA prepared in example 1 of the present invention, which shows that AM @ CS/DOX @ HA has drug release effect.
Example 2
(1) Preparing a chitosan solution: 20mg of chitosan was weighed and dissolved in 10mL of acetic acid to prepare a chitosan solution of 2mg/mL, which was stored in a refrigerator at 4 ℃.
(2) Preparing a hyaluronic acid solution: 20mg of hyaluronic acid is weighed and dissolved in 10mL of deionized water to prepare a hyaluronic acid solution of 2mg/mL, and the hyaluronic acid solution is stored in a refrigerator at 4 ℃.
(3) Preparing an adriamycin solution: 20mg of ICG was weighed and dissolved in 10mL of deionized water to prepare a 2mg/mL ICG solution, which was then sealed with tinfoil and stored in a refrigerator at 4 ℃.
(4) Preparation of AMNPs: dispersing 4g of antimony powder in a clean beaker filled with 80ml of NMP at room temperature, sealing a preservative film, carrying out ice bath ultrasonic treatment for 10 hours by using a phi 6 ultrasonic probe under an ultrasonic cell crushing instrument, wherein the power is 1000w, the ultrasonic treatment is stopped for 1s every 3s, stirring a dispersion liquid by using a glass rod every 30min to uniformly disperse the dispersion liquid, simultaneously observing whether ice blocks are dissolved in the ice bath, supplementing the ice blocks in time, carrying out ice bath ultrasonic treatment for 3 hours in an ultrasonic cleaner, wherein the power is 300w, and then carrying out ultrasonic treatment for 3 hours by using the ultrasonic cell crushing instrument; and finally, centrifuging: centrifuging the obtained dispersion at 3000rpm for 15min at 10 ℃, discarding deposited blocky antimony, retaining supernatant, centrifuging the supernatant at 12000rpm for 30min, collecting precipitate, adding 15ml of deionized water to obtain the stibene nano-particle dispersion, and storing at 4 ℃.
After being diluted properly, the absorbance of the AMNPs dispersion liquid under the wavelength of 808nm is measured by an ultraviolet-visible light analyzer, the concentration of antimony in the AMNPs dispersion liquid prepared by the method is calculated and obtained according to an antimony standard curve, and the AMNPs dispersion liquid with the concentration of 8mg/mL is obtained after being diluted by 4 times and is used for subsequent experiments.
(5) CS and DOX coated AMNPs: adding 1mL of chitosan solution and 1mL of DOX into 1mL of AMNPs dispersion liquid, magnetically stirring for 12h, centrifuging at the rotating speed of 12000rpm for 5min, collecting precipitates, adding 2mL of deionized water ultrasonic cleaner, and ultrasonically dispersing for 10min to obtain AMNPs @ CS-DOX nanoparticle dispersion liquid.
(6) Coated Hyaluronic Acid (HA): adding 1mL of hyaluronic acid solution into the obtained nanoparticle dispersion liquid, performing ultrasonic treatment for 20min by using an ultrasonic cleaner, performing magnetic stirring for 5h, aging overnight, centrifuging at 12000rpm for 5min, collecting precipitate, adding 2mL of deionized water, performing ultrasonic dispersion for 10min, centrifuging again according to the same centrifugation parameters, collecting precipitate, adding 2mL of deionized water, finally obtaining 1mg/mL of drug-loaded nanoparticle AM @ CS/DOX @ HA dispersion liquid, and refrigerating in a refrigerator at 4 ℃.
Example 3
(1) Preparing a chitosan solution: 15mg of chitosan was weighed and dissolved in 10mL of acetic acid to prepare a 1.5mg/mL chitosan solution, which was stored in a refrigerator at 4 ℃.
(2) Preparing a hyaluronic acid solution: weighing 15mg of hyaluronic acid, dissolving in 10mL of deionized water to prepare 1.5mg/mL hyaluronic acid solution, and storing in a refrigerator at 4 ℃.
(3) Preparing an adriamycin solution: 15mg of ICG was weighed and dissolved in 10mL of deionized water to prepare a 1.5mg/mL doxorubicin solution, which was then stored in a refrigerator at 4 ℃ with a foil envelope.
(4) Preparation of AMNPs: 3g of antimony powder is taken and dispersed in a clean beaker filled with 50ml of NMP at room temperature, a preservative film is sealed, a phi 6 ultrasonic probe is used for carrying out ice bath ultrasonic treatment for 8.5 hours under an ultrasonic cell crushing instrument, the power is 1000w, 1s is stopped every 3s of ultrasonic treatment, a glass rod is used for stirring dispersion liquid every 30min to ensure that the dispersion liquid is uniformly dispersed, whether ice blocks are dissolved in ice bath is observed at the same time, the ice blocks are supplemented in time, then ice bath ultrasonic treatment is carried out for 2.5 hours in an ultrasonic cleaner, the power is 300w, and then the ultrasonic cell crushing instrument is used for carrying out ultrasonic treatment for 2.5 hours; and finally, centrifuging: centrifuging the obtained dispersion liquid at the rotating speed of 3000rpm for 13min at the temperature of 8 ℃, discarding deposited blocky antimony, retaining supernatant, centrifuging the supernatant at the rotating speed of 10000rpm for 35min, collecting precipitate, adding 10ml of deionized water to obtain stibene nanoparticle dispersion liquid, and placing the stibene nanoparticle dispersion liquid at the temperature of 4 ℃ for storage.
After being properly diluted, the AMNPs dispersion liquid is measured to have the absorbance under the wavelength of 808nm by an ultraviolet-visible light analyzer, the concentration of antimony in the AMNPs dispersion liquid prepared by the method is calculated to be 4.5mg/mL according to an antimony standard curve, and the AMNPs dispersion liquid with the concentration of 1.5mg/mL is obtained after being diluted by 3 times and is used for subsequent experiments.
(5) CS and DOX coated AMNPs: adding 1mL of chitosan solution and 1mL of DOX into 1mL of AMNPs dispersion liquid, magnetically stirring for 10h, centrifuging at the rotating speed of 11000rpm for 4min, collecting precipitates, adding 1.5mL of deionized water ultrasonic cleaner, and ultrasonically dispersing for 10min to obtain AMNPs @ CS-DOX nano-particle dispersion liquid.
(6) Coated Hyaluronic Acid (HA): adding 1mL of hyaluronic acid solution into the obtained nanoparticle dispersion liquid, carrying out ultrasonic treatment for 17min by using an ultrasonic cleaner, carrying out magnetic stirring for 4h, aging overnight, centrifuging for 4min at the rotating speed of 11000rpm, collecting precipitate, adding 1.5mL of deionized water, carrying out ultrasonic dispersion for 10min, centrifuging again with the same centrifugal parameters, collecting precipitate, adding 1.5mL of deionized water, finally obtaining 1mg/mL of drug-loaded nanoparticle AM @ CS/DOX @ HA dispersion liquid, and refrigerating in a refrigerator at 4 ℃.
The raw materials and equipment used in the invention are common raw materials and equipment in the field if not specified; the methods used in the present invention are conventional in the art unless otherwise specified.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and all simple modifications, alterations and equivalents of the above embodiments according to the technical spirit of the present invention are still within the protection scope of the technical solution of the present invention.
Claims (4)
1. A preparation method of a tumor targeted drug-loaded nanoparticle based on stibene is characterized by comprising the following steps:
(1) Preparing a chitosan solution: dissolving chitosan in acetic acid to prepare a chitosan solution of 1-2mg/mL, and refrigerating for later use;
(2) Preparing a hyaluronic acid solution: dissolving hyaluronic acid in deionized water to prepare hyaluronic acid with concentration of 1-2mg/mL, and refrigerating for later use;
(3) Preparation of adriamycin solution: dissolving adriamycin in deionized water to prepare 1-2mg/mL adriamycin solution, and refrigerating the adriamycin solution for later use after being encapsulated by tinfoil;
(4) Preparation of AMNPs: dispersing 2-4g of antimony powder in 40-80mL of N-methyl-2-pyrrolidone, sealing a container, carrying out ice bath ultrasonic treatment by using an ultrasonic probe through an ultrasonic cell crushing instrument, observing the temperature of the ice bath at an interval of 20-40min, continuing ultrasonic treatment after stirring, and carrying out ice bath ultrasonic treatment in an ultrasonic cleaner; finally, ultrasonic treatment is carried out by an ultrasonic cell crusher;
centrifuging the obtained dispersion liquid at 8-10 ℃, removing deposited blocky antimony, centrifuging again, collecting precipitates, adding 10-15mL of deionized water to obtain an antimonene nanoparticle dispersion liquid, and refrigerating for later use;
the first centrifugation rotating speed is 2000-4000rpm, and the centrifugation time is 10-15min; the second centrifugation rotating speed is 8000-12000rpm, and the centrifugation time is 30-35min; the centrifugation temperature is not more than 10 ℃; the specification of the ultrasonic probe is phi 6, the ice-bath ultrasonic time is 8-10h, the power is 800-1200w, and the ultrasonic operation is stopped for 1s every 2-4 s; the ultrasonic cleaner is used for carrying out ice bath and ultrasonic treatment for 2-3h and has the power of 200-400w; the ultrasonic time of the ultrasonic cell crushing instrument is 2-3h;
(5) Diluting the stibene nano-particle dispersion liquid obtained in the step (4), measuring the concentration by using an ultraviolet-visible spectrophotometer, and preparing an AMNPs dispersion liquid with the concentration of 1-2mg/mL after dilution;
(6) Chitosan and doxorubicin coated AMNPs: adding 0.5-1mL of chitosan solution and 0.5-1mL of adriamycin solution into 0.5-1mL of AMNPs dispersion liquid, stirring, centrifuging, collecting precipitate, and adding 1-2mL of deionized water to obtain AMNPs @ CS-DOX nanoparticle dispersion liquid;
(7) Coating hyaluronic acid: and adding 1-2mL of hyaluronic acid solution into the obtained nanoparticle dispersion liquid, performing ultrasonic treatment in an ultrasonic cleaner, stirring, aging overnight, centrifuging twice, collecting precipitates, and finally obtaining the tumor-targeted drug-loaded nanoparticles based on the stibene.
2. The preparation method of the stibene-based tumor-targeted drug-loaded nanoparticle according to claim 1, wherein the refrigeration temperature in the steps (1) to (4) is 1-5 ℃.
3. The preparation method of the stibene-based tumor-targeted drug-loaded nanoparticle according to claim 1, wherein in the step (6), the stirring time is 8-12h; the centrifugal speed is 10000-12000rpm, and the centrifugal time is 3-5min.
4. The preparation method of the stibene-based tumor-targeted drug-loaded nanoparticle as claimed in claim 1, wherein in step (7), the ultrasound time is 15-20min, the magnetic stirring time is 3-5h, and the aging is carried out overnight; the two-time centrifugation rotating speed is 10000-12000rpm, and the time of each centrifugation is 3-5min.
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