CN110585447B - Preparation method and application of aggregated luminescent nano-particle material capable of being used as sound sensitizer - Google Patents
Preparation method and application of aggregated luminescent nano-particle material capable of being used as sound sensitizer Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0028—Disruption, e.g. by heat or ultrasounds, sonophysical or sonochemical activation, e.g. thermosensitive or heat-sensitive liposomes, disruption of calculi with a medicinal preparation and ultrasounds
- A61K41/0033—Sonodynamic cancer therapy with sonochemically active agents or sonosensitizers, having their cytotoxic effects enhanced through application of ultrasounds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0063—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres
- A61K49/0069—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form
- A61K49/0076—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form dispersion, suspension, e.g. particles in a liquid, colloid, emulsion
- A61K49/0084—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form dispersion, suspension, e.g. particles in a liquid, colloid, emulsion liposome, i.e. bilayered vesicular structure
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
The invention discloses a preparation method and application of an aggregated luminescent nano-particle material capable of being used as a sound-sensitive agent, which comprises the following steps: (1) weighing tetra [ tetra- (4-aminophenyl) ] phenylethene (TAPE) and dissolving in chloroform to obtain a solution A; weighing liposome and dissolving the liposome in chloroform to obtain a solution B; adding the solution A into the solution B; (2) removing chloroform by rotary evaporation at normal temperature to obtain a TAPE-lipid nanoparticle lipid membrane C; (3) adding ultrapure water into a round-bottom flask to dissolve a TAPE-lipid nanoparticle lipid membrane C; (4) placing the mixture in an ultrasonic cell disruptor for ultrasonic treatment. The nano-particles prepared by the method solve the application of water-soluble AIE materials in organisms, develop novel functions, combine imaging function and ultrasonic activation function, realize the diagnosis-treatment integrated function of tumors, and have important application prospects in the fields of nano-medicine, disease diagnosis, treatment for overcoming the limitation of tumor hypoxia and the like.
Description
Technical Field
The invention relates to the technical field of nano medicine. In particular to a preparation method of an aggregated luminescent nano-particle material which can be used as a sound-sensitive agent.
Background
The sonodynamic therapy (SDT) uses ultrasonic to excite the sonosensitizer, and in the process of acoustic cracking caused by sonoluminescence, energy is transferred to oxygen molecules or water molecules to generate high-activity singlet oxygen, free radicals and other Reactive Oxygen Species (ROS) to kill tumor cells, so as to achieve the purpose of treating tumors. The sonodynamic therapy has the advantages of deep penetration, non-invasion, space-time controllability and the like, and becomes a research hotspot in the field of tumor treatment in recent years, and the research and development of the sonosensitizer are the key points of the development of the electrokinetic therapy.
Tetrakis [ tetrakis- (4-aminophenyl) ] phenylethene (TAPE) of the formula:
as a tetraphenylethylene derivative, a typical AIE fluorescent luminophore, which has a propeller-shaped structure with a freely rotatable peripheral benzene ring, is widely used in many important fields such as electrical conduction research, functional materials, sensing, imaging, etc., however, the application in the field of disease treatment is still very lacking. In addition, poor water solubility is a major barrier to its widespread study in the fields of biological research and disease treatment. Therefore, the design of biocompatible nanoparticles and the development of new functions of the biocompatible nanoparticles in the therapeutic field provide ideas for the research and application fields of AIE materials.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to provide a preparation method of an aggregation luminescent nanoparticle material which has good water solubility and aggregation luminescent function and can be used as a sound sensitive agent.
In order to solve the technical problems, the invention provides the following technical scheme:
a method for preparing aggregation luminous nano-particle material used as a sound-sensitive agent adopts a thin film method to prepare nano-particles, and comprises the following steps:
(1) weighing tetra [ tetra- (4-aminophenyl) ] phenylethene, and dissolving in chloroform to obtain a solution A; weighing liposome and dissolving the liposome in chloroform to obtain a solution B; adding the solution A into the solution B, and stirring at normal temperature until the solution A is dissolved;
(2) at normal temperature, rotationally evaporating by using a rotary evaporator under a vacuum condition to remove chloroform, adjusting a circulating water type multipurpose vacuum pump to the maximum vacuum degree when no solvent is observed in the round-bottom flask, and then rotationally evaporating for a certain time till the mixture is completely dried to obtain a TAPE-lipid nanoparticle lipid membrane C;
(3) adding ultrapure water into a round-bottom flask to dissolve the TAPE-lipid nanoparticle lipid membrane C, and transferring the dissolved TAPE-lipid nanoparticle lipid membrane C into a centrifuge tube;
(4) and (3) placing the centrifugal tube into an ultrasonic cell disruptor for ultrasonic treatment to obtain TAPE-lipid suspension D, filtering by using an ultrafiltration membrane to obtain TAPE-lipid nanoparticle solution, and storing in a refrigerator at 4 ℃ for later use.
In the above method for preparing aggregated luminescent nanoparticle material for use as a sonosensitizer, in step (1), the mass ratio of the tetrakis [ tetrakis- (4-aminophenyl) ] phenylethene to the liposome is 1: 5-20.
In the step (2), when no solvent is observed in the round-bottom flask, the circulating water type multipurpose vacuum pump is adjusted to the maximum vacuum degree, and then the mixture is steamed for 2-10min in a rotary manner, and then the mixture is steamed to be completely dry at the temperature of 45 ℃.
In the step (3), 1-10mL of ultrapure water is added into a bottle to dissolve the aggregated luminescent nanoparticle material which can be used as the sound-sensitive agent, and then the solution is transferred into a 10-50mL centrifuge tube.
In the step (4), the rated power of the ultrasonic cell disruption instrument is 150W, the frequency is 20kHz, the ultrasonic power is 20% -90% of the rated power, and the ultrasonic time is 3-10 min.
The preparation method of the aggregation luminescent nano-particle material which can be used as the sound-sensitive agent comprises the following steps of (1) preparing a liposome which is a mixture of soybean lecithin and cholesterol; the mass ratio of the soybean lecithin to the cholesterol is as follows: soybean lecithin: cholesterol (5-3): 1.
in the preparation method of the aggregated luminescent nanoparticle material capable of being used as the sonosensitizer, 3.6mg of TAPE is weighed and dissolved in 1mL of chloroform to obtain a solution A; dissolving 54mg of soybean lecithin and 18mg of cholesterol in 20mL of chloroform to obtain a mixed solution B; adding the solution A into the mixed solution B; stirring at normal temperature to dissolve, placing in a rotary evaporator, rotary evaporating at normal temperature to remove organic solvent, observing no solvent in the bottle, rotating at maximum vacuum degree for 5min, and rotating at 45 deg.C to completely dry to obtain TAPE-lipid nanoparticle lipid membrane C; adding 4mL of ultrapure water into a bottle to dissolve the ultrapure water, transferring the dissolved ultrapure water into a 10mL centrifuge tube, and carrying out ultrasonic hydration for 15min in ultrasonic cleaning waves to obtain a TAPE liposome suspension D; the rated power of the ultrasonic cell disruptor is 150W, the frequency is 20kHz, the ultrasonic power is 30 percent of the rated power, the time is 5min, after ultrasonic treatment, the TAPE-lipid nanoparticle solution is obtained after the filtration by an ultrafiltration membrane with the aperture of 0.22 μm.
In the preparation method of the aggregated luminescent nanoparticle material capable of being used as the sonosensitizer, 3.6mg of TAPE is weighed and dissolved in 1mL of chloroform to obtain a solution A; dissolving 54mg of soybean lecithin and 18mg of cholesterol in 10mL of chloroform to obtain a mixed solution B; adding the solution A into the mixed solution B; stirring at normal temperature to dissolve, placing in a rotary evaporator, rotary evaporating at normal temperature to remove organic solvent, observing no solvent in the bottle, rotating at maximum vacuum degree for 5min, and rotating at 45 deg.C to completely dry to obtain TAPE-lipid nanoparticle lipid membrane C; adding 3mL of ultrapure water into a bottle, dissolving the ultrapure water, transferring the dissolved ultrapure water into a 10mL centrifuge tube, and carrying out ultrasonic hydration for 15min in ultrasonic cleaning waves to obtain a TAPE liposome suspension D; the rated power of the ultrasonic cell disruptor is 150W, the frequency is 20kHz, the ultrasonic power is 30 percent of the rated power, the time is 5min, after ultrasonic treatment, the TAPE-lipid nanoparticle solution is obtained after the filtration by an ultrafiltration membrane with the aperture of 0.22 μm.
In the preparation method of the aggregated luminescent nanoparticle material capable of being used as the sonosensitizer, 3.6mg of TAPE is weighed and dissolved in 1mL of chloroform to obtain a solution A; dissolving 54mg of soybean lecithin and 18mg of cholesterol in 20mL of chloroform to obtain a mixed solution B; adding the solution A into the mixed solution B; stirring at normal temperature to dissolve, placing in a rotary evaporator, rotary evaporating at normal temperature to remove organic solvent, observing no solvent in the bottle, rotating at maximum vacuum degree for 5min, and rotating at 45 deg.C to completely dry to obtain TAPE-lipid nanoparticle lipid membrane C; adding 4mL of ultrapure water into a bottle to dissolve the ultrapure water, transferring the dissolved ultrapure water into a 10mL centrifuge tube, and carrying out ultrasonic hydration for 15min in ultrasonic cleaning waves to obtain a TAPE liposome suspension D; the rated power of the ultrasonic cell disruptor is 150W, the frequency is 20kHz, the ultrasonic power is 30 percent of the rated power, the time is 5min, after ultrasonic treatment, the TAPE-lipid nanoparticle solution is obtained after the filtration by an ultrafiltration membrane with the aperture of 0.22 μm.
Use of an aggregated luminescent nanoparticle material as a sonosensitizer in the sonodynamic treatment of tumours.
The technical scheme of the invention achieves the following beneficial technical effects:
1. the application of tetra [ tetra- (4-aminophenyl) ] phenylethylene in organisms is severely limited due to poor water solubility of the tetra [ tetra- (4-aminophenyl) ] phenylethylene serving as a typical AIE material, and the nano particles are prepared from the tetra [ tetra- (4-aminophenyl) ] phenylethylene and an amphiphilic carrier, so that the solubility of the tetra [ tetra- (4-aminophenyl) ] phenylethylene in water is increased, and the application value of the tetra [ tetra- (4-aminophenyl) ] phenylethylene in organisms is improved; the key point of the invention is that the problem of poor water solubility of the aggregated luminescent material in the prepared nano liposome particles is overcome, and the invention is more beneficial to being applied to imaging probes in organisms.
2. Fluorescence imaging shows that the aqueous solution of the nano-particles has an obvious fluorescence signal near 800nm and can be used as a fluorescence probe for fluorescence imaging in a living body; in addition, the fluorescence intensity of the nano-particles shows an enhanced trend along with the increase of concentration, and the function of gathering and emitting light is beneficial to the detection of drugs in organisms and the function of lightening the focal tissues of organisms, thereby providing a basis for monitoring the drug enrichment and treatment in real time.
3. Further ultrasonic excitation shows that the nano-particles can also generate singlet oxygen under the ultrasonic excitation, and the singlet oxygen has the function of killing tumor cells, so the nano-particles can be used as a sonosensitizer and have the function of dynamically treating tumors, and the property provides a basis for the nano-particles to serve as a diagnosis-treatment integrated biological material.
The nano-particle solves the application of the water-soluble AIE material in organisms, develops novel functions simultaneously, combines an imaging function and an ultrasonic activation function, can realize the diagnosis-treatment integrated function of tumors, and has important application prospect in the fields of nano-medicine, disease diagnosis, treatment for overcoming the limitation of tumor hypoxia and the like.
Drawings
FIG. 1 is a synthesis scheme of a method for preparing aggregated luminescent nanoparticle materials useful as sonosensitizers according to the present invention;
FIG. 2 is an electron microscope image of TAPE-Lipid nanoparticles (tetrakis [ tetrakis- (4-aminophenyl) ] phenylethene-liposome nanoparticles) obtained by the preparation method of the aggregated luminescent nanoparticle material used as a sonosensitizer of the present invention;
FIG. 3 is a graph showing the increase in fluorescence intensity of tetrakis [ tetrakis- (4-aminophenyl) ] phenylethenTAPE-liposome nanoparticles with increasing concentration;
FIG. 4 shows a quenching diagram of the absorption value of DPBF due to singlet oxygen generated by the nanoparticles after ultrasonic excitation.
Detailed Description
First part, preparation of aggregated luminescent nanoparticle materials useful as sonosensitizers (tetrakis [ tetrakis- (4-aminophenyl)]Benzene and its derivatives
Vinyl TAPE-Liposome nanoparticle)
Example 1:
preparing TAPE-liposome nanoparticles: dissolving 3.6mg of TAPE in 1mL of chloroform to obtain a solution A; dissolving 36mg of soybean lecithin and 12mg of cholesterol in 10mL of chloroform to obtain a mixed solution B; adding the solution A into the mixed solution B. Stirring at normal temperature to dissolve, placing in a rotary evaporator, performing rotary evaporation at normal temperature under vacuum condition to remove chloroform, adjusting the maximum vacuum degree (minus 0.098Mpa) of a circulating water type multi-purpose vacuum pump when no solvent is in the observation bottle, rotating for 5min, and rotating at 45 deg.C to completely dry to obtain TAPE-lipid nanoparticle lipid membrane C. And adding 2mL of ultrapure water into the bottle to dissolve the liposome, transferring the dissolved liposome into a 10mL centrifuge tube, and carrying out ultrasonic hydration for 15min in ultrasonic cleaning waves to obtain a TAPE liposome suspension D. The rated power of an ultrasonic cell disruptor (sonic & metals inc, USA) is 150W, the frequency is 20kHz, the ultrasonic power is 30 percent of the rated power, the time is 5min, after ultrasonic treatment, the TAPE-lipid nanoparticle solution is obtained after filtering by an ultrafiltration membrane with the aperture of 0.22 mu m.
Example 2:
preparing TAPE-liposome nanoparticles: weighing 3.6mg of TAPE, and dissolving in 1mL of chloroform to obtain a solution A; dissolving 54mg of soybean lecithin and 18mg of cholesterol in 10mL of chloroform to obtain a mixed solution B; adding the solution A into the mixed solution B. Stirring at normal temperature to dissolve, placing in a rotary evaporator, performing rotary evaporation at normal temperature under vacuum condition to remove chloroform, adjusting the maximum vacuum degree (minus 0.098Mpa) of a circulating water type multi-purpose vacuum pump when no solvent is in the observation bottle, rotating for 5min, and rotating at 45 deg.C to completely dry to obtain TAPE-lipid nanoparticle lipid membrane C. And adding 3mL of ultrapure water into the bottle to dissolve the mixture, transferring the dissolved mixture into a 10mL centrifuge tube, and carrying out ultrasonic hydration for 15min in ultrasonic cleaning waves to obtain the TAPE liposome suspension D. The rated power of an ultrasonic cell disruptor (sonic & metals inc, USA) is 150W, the frequency is 20kHz, the ultrasonic power is 30 percent of the rated power, the time is 5min, after ultrasonic treatment, the TAPE-lipid nanoparticle solution is obtained after filtering by an ultrafiltration membrane with the aperture of 0.22 mu m.
Example 3:
preparing TAPE-liposome nanoparticles: weighing 3.6mg of TAPE, and dissolving in 1mL of chloroform to obtain a solution A; dissolving 54mg of soybean lecithin and 18mg of cholesterol in 20mL of chloroform to obtain a mixed solution B; adding the solution A into the mixed solution B. Stirring at normal temperature to dissolve, placing in a rotary evaporator, performing rotary evaporation at normal temperature under vacuum condition to remove organic solvent, adjusting the maximum vacuum degree (minus 0.098Mpa) of a circulating water type multi-purpose vacuum pump when no solvent is in the observation bottle, rotating for 5min, and rotating at 45 deg.C to completely dry to obtain TAPE-lipid nanoparticle lipid membrane C. Adding 4mL of ultrapure water into a bottle to dissolve the liposome, transferring the dissolved liposome into a 10mL centrifuge tube, and carrying out ultrasonic hydration for 15min in an ultrasonic cleaning wave to obtain a TAPE liposome suspension D. The rated power of an ultrasonic cell disruptor (sonic & metals inc, USA) is 150W, the frequency is 20kHz, the ultrasonic power is 30 percent of the rated power, the time is 5min, after ultrasonic treatment, the TAPE-lipid nanoparticle solution is obtained after filtering by an ultrafiltration membrane with the aperture of 0.22 mu m.
FIG. 1 is a schematic diagram of the synthesis of TAPE-doped nanoparticles in examples 1 to 3, and examples 1 to 3 all can obtain uniform-sized nanoparticles (as shown in FIG. 2) and uniform spherical nanoparticles with a particle size of about 60 nm.
Second moiety, tetrakis [ tetrakis- (4-aminophenyl)]Fluorescence characteristic and application of phenylethene-liposome nanoparticles
The TAPE-Lipid nanoparticle solution prepared in example 3 was diluted to concentrations of 2. mu.g/mL, 4. mu.g/mL, 8. mu.g/mL, 16. mu.g/mL, 32. mu.g/mL and 64. mu.g/mL (TAPE concentration) by a stepwise dilution method, and fluorescence spectra of the solutions at respective concentrations were measured in descending order of concentration to confirm a change curve of fluorescence intensity versus concentration. As shown in fig. 3, the maximum emission spectrum of the nanoparticle is around 800nm, and meanwhile, the fluorescence intensity gradually increases with the increase of the concentration, so that the nanoparticle has the characteristic of obvious aggregation enhancement, and this property can be used for imaging the lesion tissue in vivo, and as the nanoparticle is aggregated in the lesion tissue, the lesion site becomes clearer and more obvious, so that the nanoparticle has a good imaging diagnosis function.
Detecting singlet oxygen generated by the nanoparticle solution under ultrasonic excitation: diluting the TAPE-Lipid nanoparticle solution prepared in example 3 to 50 ug/mL; taking 3mL of the diluted solution in a culture dish with the diameter of 2.5cm, and marking as a solution E; dissolving a 1, 3-diphenyl isobenzofuran ethanol solution (DPBF, a singlet oxygen quencher) into a 0.5mM solution by using ethanol, putting 10uLDPBF solution into an E solution, uniformly mixing, continuously ultrasonically exciting for 10-80 seconds by using an ultrasonic therapeutic apparatus, taking 50 microliters of the solution every 10 seconds, diluting to 1mL, and detecting the change condition of the absorption value of the solution at the position of 420nm by using an ultraviolet spectrophotometer. As can be seen from fig. 4, with the increasing ultrasonic excitation time, the absorption value near 420nm decreases, indicating that singlet oxygen is generated in the ultrasonic excitation; singlet oxygen combines with DPBF, resulting in a decrease in absorbance and an increasing production of singlet oxygen over time, and is useful as a sonosensitizer for sonodynamic therapy.
Therefore, the TAPE-Lipid nanoparticle prepared by the invention has the integrated capability of diagnosis and treatment of lesion tissues such as tumor.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications are possible which remain within the scope of the appended claims.
Claims (9)
1. A method for preparing an aggregated luminescent nanoparticle material which can be used as a sound-sensitive agent is characterized in that a thin film method is adopted to prepare nanoparticles, and the method comprises the following steps:
(1) weighing tetra [ tetra- (4-aminophenyl) ] phenylethene, and dissolving in chloroform to obtain a solution A; weighing liposome and dissolving the liposome in chloroform to obtain a solution B; adding the solution A into the solution B, and stirring at normal temperature until the solution A is dissolved;
(2) at normal temperature, rotationally evaporating by using a rotary evaporator under a vacuum condition to remove chloroform, adjusting a circulating water type multipurpose vacuum pump to the maximum vacuum degree when no solvent is observed in the round-bottom flask, and then rotationally evaporating for a certain time till the mixture is completely dried to obtain a TAPE-lipid nanoparticle lipid membrane C;
(3) adding ultrapure water into a round-bottom flask to dissolve the TAPE-lipid nanoparticle lipid membrane C, and transferring the dissolved TAPE-lipid nanoparticle lipid membrane C into a centrifuge tube;
(4) and (3) placing the centrifugal tube into an ultrasonic cell disruptor for ultrasonic treatment to obtain TAPE-lipid suspension D, filtering by using an ultrafiltration membrane to obtain TAPE-lipid nanoparticle solution, and storing in a refrigerator at 4 ℃ for later use.
2. The method for preparing an aggregated luminescent nanoparticle material useful as a sonosensitizer according to claim 1, wherein in step (1), the mass ratio of said tetrakis [ tetrakis- (4-aminophenyl) ] phenylethene to said liposome is 1: 5-20.
3. The method for preparing the aggregated luminescent nanoparticle material used as a sonosensitizer of claim 1, wherein in the step (2), when the round-bottomed flask is observed to be free of solvent, the vacuum pump with circulating water type is adjusted to the maximum vacuum degree, and then the mixture is spin-evaporated for 2-10min, and then the mixture is spin-evaporated to be completely dry at 45 ℃.
4. The method for preparing an aggregated luminescent nanoparticle material capable of being used as a sonosensitizer according to claim 1, wherein in step (3), 1-10mL of ultrapure water is added into a bottle to be dissolved, and then the dissolved ultrapure water is transferred into a 10-50mL centrifuge tube.
5. The method for preparing the aggregated luminescent nanoparticle material that can be used as a sonosensitizer of claim 1, wherein in step (4), the power rating of the ultrasonic cell disruptor is 150W, the frequency is 20kHz, the ultrasonic power is 20% -90% of the power rating, and the ultrasonic time is 3-10 min.
6. The method for preparing an aggregated luminescent nanoparticle material useful as a sonosensitizer according to any one of claims 1-5, wherein said liposome is a mixture of soy lecithin and cholesterol; the mass ratio of the soybean lecithin to the cholesterol is as follows: soybean lecithin: cholesterol = (5-3): 1.
7. the method for preparing an aggregated luminescent nanoparticle material useful as a sonosensitizer of claim 6, wherein 3.6mg of TAPE is weighed and dissolved in 1mL of chloroform to obtain a solution A; dissolving 54mg of soybean lecithin and 18mg of cholesterol in 20mL of chloroform to obtain a mixed solution B; adding the solution A into the mixed solution B; stirring at normal temperature to dissolve, placing in a rotary evaporator, rotary evaporating at normal temperature to remove organic solvent, observing no solvent in the bottle, rotating at maximum vacuum degree for 5min, and rotating at 45 deg.C to completely dry to obtain TAPE-lipid nanoparticle lipid membrane C; adding 4mL of ultrapure water into a bottle to dissolve the ultrapure water, transferring the dissolved ultrapure water into a 10mL centrifuge tube, and carrying out ultrasonic hydration for 15min in ultrasonic cleaning waves to obtain a TAPE liposome suspension D; the rated power of the ultrasonic cell disruptor is 150W, the frequency is 20kHz, the ultrasonic power is 30 percent of the rated power, the time is 5min, after ultrasonic treatment, the TAPE-lipid nanoparticle solution is obtained after the filtration by an ultrafiltration membrane with the aperture of 0.22 μm.
8. The method for preparing an aggregated luminescent nanoparticle material useful as a sonosensitizer of claim 6, wherein 3.6mg of TAPE is weighed and dissolved in 1mL of chloroform to obtain a solution A; dissolving 54mg of soybean lecithin and 18mg of cholesterol in 10mL of chloroform to obtain a mixed solution B; adding the solution A into the mixed solution B; stirring at normal temperature to dissolve, placing in a rotary evaporator, rotary evaporating at normal temperature to remove organic solvent, observing no solvent in the bottle, rotating at maximum vacuum degree for 5min, and rotating at 45 deg.C to completely dry to obtain TAPE-lipid nanoparticle lipid membrane C; adding 3mL of ultrapure water into a bottle, dissolving the ultrapure water, transferring the dissolved ultrapure water into a 10mL centrifuge tube, and carrying out ultrasonic hydration for 15min in ultrasonic cleaning waves to obtain a TAPE liposome suspension D; the rated power of the ultrasonic cell disruptor is 150W, the frequency is 20kHz, the ultrasonic power is 30 percent of the rated power, the time is 5min, after ultrasonic treatment, the TAPE-lipid nanoparticle solution is obtained after the filtration by an ultrafiltration membrane with the aperture of 0.22 μm.
9. Use of aggregated luminescent nanoparticle material, characterized in that the aggregated luminescent nanoparticle material prepared according to any of claims 1 to 8 is used for the preparation of a sonosensitizer.
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| WO2012143739A1 (en) * | 2011-04-21 | 2012-10-26 | University Of Ulster | Sonodynamic therapy |
| CN110025576A (en) * | 2019-04-23 | 2019-07-19 | 上海市第六人民医院 | A kind of preparation method and applications of the photothermal reagent of the photo-thermal oncotherapy mediated for fluorescence imaging |
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| WO2012143739A1 (en) * | 2011-04-21 | 2012-10-26 | University Of Ulster | Sonodynamic therapy |
| CN110025576A (en) * | 2019-04-23 | 2019-07-19 | 上海市第六人民医院 | A kind of preparation method and applications of the photothermal reagent of the photo-thermal oncotherapy mediated for fluorescence imaging |
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