CN114246947B

CN114246947B - Inorganic sound sensitizer independent of oxygen in tumor microenvironment, preparation method and application

Info

Publication number: CN114246947B
Application number: CN202111618253.1A
Authority: CN
Inventors: 裴仁军; 孙丽娜
Original assignee: Suzhou Institute of Nano Tech and Nano Bionics of CAS
Current assignee: Suzhou Institute of Nano Tech and Nano Bionics of CAS
Priority date: 2021-12-27
Filing date: 2021-12-27
Publication date: 2023-05-23
Anticipated expiration: 2041-12-27
Also published as: CN114246947A

Abstract

The invention discloses an inorganic sound sensitizer which does not depend on oxygen in a tumor microenvironment. The inorganic sound sensitizer is metal doped manganese oxide nano particles, and because of the abundant oxygen vacancy structure, a large number of oxygen molecules can be spontaneously adsorbed on the surface, the oxygen molecules can be provided for an oxygen source rich in the sound sensitizer in a hypoxia state, and after the sound sensitizer is irradiated by ultrasound, the oxygen molecules adsorbed on the surface are converted into singlet oxygen with cytotoxicity, so that the tumor cells are killed. The sound-sensitive agent provided by the invention can be free from limitation of hypoxia in a tumor microenvironment, and can exert a good sound power treatment effect. The work provides new insights and ideas for developing high-efficiency inorganic sound-sensitive agents which do not depend on oxygen in tumor microenvironment.

Description

Inorganic sound sensitizer independent of oxygen in tumor microenvironment, preparation method and application

Technical Field

The invention relates to an acoustic sensitizer, in particular to an inorganic acoustic sensitizer independent of oxygen in a tumor microenvironment, and a preparation method and application thereof, belonging to the technical field of functional inorganic nano materials.

Background

Traditional tumor treatment schemes, such as radiotherapy and chemotherapy, have lower efficiency, strong toxic and side effects, tolerance and the like. The novel tumor treatment scheme has higher efficiency, is mostly noninvasive treatment, and can carry out accurate treatment. Therefore, the development of new tumor treatment regimens is particularly important.

The acoustic power treatment is a novel tumor treatment method which utilizes the ultrasonic wave with the frequency within the range of 20 kHz-3 MHz and without thermal effect to reach the deep covered soft tissue and simultaneously activates the acoustic sensitizer to generate active oxygen, thereby achieving the effect of treating tumors. His penetration depth is deeper (10 cm) than the optical treatment in either the near infrared first region (690-950 nm) or the near infrared second region (1000-1350 nm), and is of great interest. The widely accepted mechanism for active oxygen production for sonodynamic therapy is the cavitation effect caused by ultrasound generation, which initiates both sonoluminescence and thermal decomposition. The energy generated by the sonoluminescence causes the sonosensitizer to transit from a ground state to an excited state, and the energy is released in the process of returning from the excited state to the ground state, so that oxygen is converted into singlet oxygen; and the heat generated by cavitation effect can directly split water into hydroxyl radicals.

At present, the sound sensitive agent is mainly divided into an organic sound sensitive agent and an inorganic sound sensitive agent. The organic sound-sensitive agent is mainly porphyrin derivatives, is widely developed due to good biocompatibility and potential clinical application value, and generally has the defects of poor water solubility, difficult enrichment at tumor sites, too short blood circulation time in vivo and the like, so that good treatment effect cannot be exerted. Whereas inorganic sonosensitizers such as titanium dioxide generally have lower phototoxicity and better chemical stability. However, oxygen is the most important raw material for producing active oxygen, and the lack of oxygen at the tumor site can make the sonodynamic therapy effect unsatisfactory. Therefore, the development of inorganic sound-sensitive agents capable of overcoming the tumor hypoxia state is a urgent problem to be solved.

Disclosure of Invention

The invention aims to solve the problems and provide an inorganic sound sensitizer independent of oxygen in tumor microenvironment and a preparation method thereof.

It is also an object of the present invention to provide the use of said inorganic sound-sensitive agent independent of oxygen in the tumor microenvironment.

In order to achieve the aim of the invention, the invention adopts the following technical scheme:

the embodiment of the invention provides a preparation method of an inorganic sound-sensitive agent independent of oxygen in a tumor microenvironment, which comprises the following steps:

carrying out a first reaction for 2-4 hours at 70-90 ℃ in a first mixed reaction system comprising a first precursor, oleic acid, oleylamine and a high-boiling point organic solvent, wherein the first precursor is manganese acetylacetonate;

heating the obtained reaction system to 260-320 ℃ in a protective atmosphere, adding a second precursor to form a second mixed reaction system, and continuing the second reaction for 15-60 min to obtain metal doped manganese oxide nano particles, wherein the second precursor is metal acetylacetonate;

and modifying the amphiphilic polyethylene glycol on the surface of the metal-doped manganese oxide nanoparticle to obtain the inorganic sound sensitizer which does not depend on oxygen in the tumor microenvironment.

The embodiment of the invention also provides the inorganic sound-sensitive agent which is prepared by the method and does not depend on oxygen in the tumor microenvironment, wherein the inorganic sound-sensitive agent is manganese oxide nano particles with the Fe/Mn metal molar doping ratio of 6.7% -20%, the inorganic sound-sensitive agent which does not depend on oxygen in the tumor microenvironment has a rich oxygen vacancy structure, and a large number of oxygen molecules can be spontaneously adsorbed on the surface of the inorganic sound-sensitive agent.

The embodiment of the invention also provides application of the inorganic sound sensitizer independent of oxygen in the tumor microenvironment in preparation of an antibacterial product or a product for treating tumors by acoustic power.

Compared with the prior art, the invention has the beneficial effects that:

1) The abundant defects constructed by the oxygen vacancy engineering technology used in the invention can play the role of electron traps, inhibit recombination of electrons and holes, thereby obviously improving the active oxygen quantum yield of the sound sensitizer;

2) The invention introduces oxygen vacancy engineering technology to construct the sound sensitizer with abundant defects, and the surface of the sound sensitizer can adsorb a large amount of oxygen molecules, thereby providing abundant oxygen sources for sound power treatment under the condition of hypoxia. The surface of the sound sensitizer can spontaneously adsorb a large amount of oxygen molecules due to the abundant oxygen vacancy structure, and the oxygen molecules can be provided for the sound sensitizer oxygen source in the hypoxia state, so that the sound sensitizer can not be limited by hypoxia in a tumor microenvironment, and a good sound power treatment effect is exerted, the sound sensitizer is a novel high-efficiency sound sensitizer, and oxygen source can be provided by oxygen molecules adsorbed on the surface of the sound sensitizer in sound power treatment, so that the problem that the hypoxia in the tumor microenvironment restricts the sound sensitization efficiency of the sound sensitizer is solved;

3) The oxygen vacancy engineering technology adopted by the invention has universality and can be used for constructing other sound sensitizers which do not depend on oxygen in tumor microenvironment.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments or the description of the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments described in the present invention, and other drawings may be obtained according to the drawings without inventive effort to those skilled in the art.

FIG. 1 is a schematic illustration of an acoustic sensitizer and its use in an exemplary embodiment of the present invention;

FIG. 2 is an electron microscopic image of the sound-sensitive agent with independent oxygen characteristics in tumor microenvironment prepared in example 1 of the present invention;

FIG. 3 is a graph showing the particle size distribution of an acoustic sensitizer having oxygen independent characteristics in tumor microenvironment prepared in example 1 of the present invention;

FIG. 4 is a graph showing the ultraviolet absorption spectrum of the acoustic sensitizer having an oxygen independent property in tumor microenvironment prepared in example 1 of the present invention;

FIG. 5 is a solid electron paramagnetic resonance spectrum of the sound-sensitive agent with independent oxygen characteristics in tumor microenvironment prepared in example 1 of the present invention;

FIG. 6 is a schematic illustration of single-line oxygen generated under acoustic power by electron spin resonance detection in the presence of a capture agent TEMP for an acoustic sensitizer having oxygen characteristics independent of the tumor microenvironment prepared in example 1 of the present invention;

FIG. 7 is a bar graph of the effect of the sonosensitizer prepared in example 1 of the present invention having oxygen independent properties in tumor microenvironment on cancer cell survival under normoxic and hypoxic conditions.

Detailed Description

Aiming at a plurality of defects in the prior art, the inventor of the present invention provides a technical scheme through long-term research and a large amount of practice, and provides a preparation method of an inorganic sound-sensitive agent which does not depend on oxygen in a tumor microenvironment. The technical scheme, the implementation process, the principle and the like are further explained as follows. It should be understood, however, that within the scope of the present invention, the above-described technical features of the present invention and technical features specifically described in the following (embodiments) may be combined with each other to constitute new or preferred technical solutions. And are limited to a space, and are not described in detail herein.

As one aspect of the technical scheme of the invention, the inorganic sound sensitizer which does not depend on oxygen in a tumor microenvironment is related, the sound sensitizer is metal doped manganese oxide nano-particles and is prepared by carrying out high-temperature pyrolysis and fusion on a first precursor and a second precursor, wherein the first precursor is a manganese doped precursor, the second precursor is a metal doped precursor, the two precursors are subjected to high-temperature pyrolysis and are fused into nano-particles, and then the surface of the nano-particles is modified with amphiphilic polyethylene glycol.

As another aspect of the technical scheme of the invention, the preparation method of the inorganic sound-sensitive agent independent of oxygen in tumor microenvironment comprises the following steps:

The preparation mechanism of the inorganic sound sensitizer independent of oxygen in tumor microenvironment is as follows: when the invention prepares the sound sensitive agent which does not depend on oxygen in tumor microenvironment, the first precursor is thermally decomposed to form seeds at high temperature, the seeds are fused with the second precursor after the second precursor is added, the second precursor further grows up to form metal doped oxide, and the final hydrophilic sound sensitive agent is formed through modification of amphiphilic polyethylene glycol. The sound sensitizer can generate a large amount of active oxygen under normoxic and hypoxic conditions, and has good therapeutic effect on solid tumors with hypoxia.

In some specific embodiments, the specific synthetic procedure for the inorganic sound-sensitive agent independent of oxygen in the tumor microenvironment is as follows:

(1) Mixing a first precursor, oleic acid, oleylamine and a high-boiling-point organic solvent, heating to 70-90 ℃, and keeping the reaction for 2-4 hours;

(2) Continuously heating the system to 260-320 ℃, adding a second precursor, and reacting for 15-60 min to obtain metal doped manganese oxide nano particles;

(3) Finally, metal doped manganese oxide nano particles precipitated by a polar solvent are centrifuged, and amphiphilic polyethylene glycol is modified on the surface to change the surface from hydrophobic to hydrophilic;

(4) The inorganic sound sensitive agent with the modified surface is used for resisting bacteria and treating tumor by sound power.

In some embodiments, the first precursor is manganese acetylacetonate.

Further, the second precursor is mainly metal acetylacetonates including, but not limited to, iron acetylacetonate, copper acetylacetonate, and the like.

In some embodiments, the molar volume ratio of the first precursor, oleic acid, and oleylamine is 0.5-1 mmol, 0.5-1 mL, and 0.5-1 mL. For example, the first precursor may be used in an amount of 0.5 to 1 mmol, the oleic acid may be used in an amount of 0.5 to 1 ml, and the oleylamine may be used in an amount of 0.5 to 1 ml.

In some embodiments, in step (1), oleic acid and oleylamine are necessary throughout the reaction.

Further, in the step (1), the high boiling point organic solvent includes at least any one of dibenzyl ether or 1-octadecene, but is not limited thereto. Benzyl ether may be used when the second reaction temperature is 260-280 ℃, and 1-octadecene is required when the second reaction temperature is 280-320 ℃.

In some embodiments, in step (2), the entire system should be anhydrous and oxygen-free, and the reaction is carried out under a protective atmosphere (e.g., nitrogen protection).

Further, in the step (2), the temperature at the time of adding the second precursor must be kept at 260 ℃ or higher, preferably 260 to 320 ℃. The molar ratio of the added second precursor to the first precursor is 5-25:100, and alternatively, the added second precursor is 5-25% of the molar amount of the first precursor.

In some embodiments, in step (3), the amphiphilic polyethylene glycol comprises at least any one of methoxy-polyethylene glycol-phospholipid or polyethylene glycol grafted polymaleic anhydride-1-octadecene.

Further, the mass ratio of the amphiphilic polyethylene glycol to the whole inorganic sound sensitizer is 5:1-10:1.

Further, step (3) includes: after the second reaction is completed, adopting a polar solvent to precipitate metal doped manganese oxide nano particles, and centrifuging, wherein the polar solvent adopted for precipitation comprises acetone, ethanol and the like; and the volume ratio of the polar solvent to the second mixed reaction system should be greater than 10:1.

In another aspect, embodiments of the present invention provide an inorganic sound-sensitive agent prepared by the foregoing method that is independent of oxygen in a tumor microenvironment.

The inorganic sound sensitizer which does not depend on oxygen in the tumor microenvironment is manganese oxide nano particles with the metal doping mole ratio (Fe/Mn) of 6.7% -20.0%, has rich oxygen vacancy structures, can spontaneously adsorb a large number of oxygen molecules on the surface, and provides rich oxygen sources for sound power treatment under the condition of hypoxia.

Furthermore, the inorganic sound sensitizer independent of oxygen in the tumor microenvironment is an ultra-small-sized metal doped manganese oxide nanoparticle, and the surface of the inorganic sound sensitizer is modified by amphiphilic polyethylene glycol so that the inorganic sound sensitizer has good water solubility.

Specifically, the particle size of the inorganic sound sensitive agent is 5-20 nm, preferably uniform particle size and particle size below 10 nm.

In another aspect, the embodiment of the invention also provides an application of the inorganic sound-sensitive agent independent of oxygen in the tumor microenvironment in preparing a product for treating tumors by sound power.

Further, the ultrasonic device used in the acoustic power therapy is selected from the parameters of20 kHz~1.0 MHz, 1.5~3.0 W/cm ² 40% -50% duty ratio.

In conclusion, the abundant defects constructed by the oxygen vacancy engineering technology used by the invention can play the role of electron traps, inhibit recombination of electrons and holes, and thus remarkably improve the active oxygen quantum yield of the sound-sensitive agent; compared with the existing inorganic sound sensitizer, the sound sensitizer prepared by the invention has rich oxygen vacancy structures, and a large number of oxygen molecules can be spontaneously adsorbed on the surface, and can be provided for an oxygen source of the sound sensitizer in a hypoxia state, so that the sound sensitizer can not be limited by hypoxia in a tumor microenvironment, and a good sound power treatment effect is exerted, and the sound sensitizer is a novel high-efficiency sound sensitizer.

The invention will now be described in detail with reference to the drawings and specific examples. The present embodiment is implemented on the premise of the technical scheme of the present invention, and a detailed implementation manner and a specific operation process are given, but the protection scope of the present invention is not limited to the following examples.

The ultrasonic device which is used in the acoustic power treatment process of the acoustic sensitizer which does not depend on the oxygen characteristic in the tumor microenvironment and is prepared by the invention preferably selects parameters of 20 kHz-1.0 MHz and 1.5-3.0W/cm ² 40% -50% duty ratio.

According to the invention, TEMP is used as a capturing agent, electron spin resonance is adopted to detect single-wire oxygen generated in acoustic power, and under ultrasonic radiation, the prepared acoustic sensitizer with the characteristic of independent oxygen in tumor microenvironment can generate abundant single-wire oxygen under normal oxygen and hypoxia conditions.

Example 1

The preparation process of the sound-sensitive agent with the characteristic of independent oxygen in tumor microenvironment in the embodiment comprises the following steps:

(1) Manganese acetylacetonate 1 mmol is mixed with oleic acid 1 ml, oleylamine 1 ml and dibenzyl ether 20 ml, heated to 70 ℃ and kept for reaction for 2 hours;

(2) Continuously heating the system to 260 ℃, adding 0.25 millimole of ferric acetylacetonate, and reacting for 15 minutes to obtain iron-doped manganese oxide nano particles;

(3) Finally precipitating iron doped manganese oxide nano particles by ethanol, centrifuging, and modifying methoxy-polyethylene glycol-phospholipid on the surface to change the surface from hydrophobic to hydrophilic. The sound sensitizer which does not depend on oxygen characteristics in tumor microenvironment is prepared, and the process schematic diagram is shown in figure 1.

Fig. 2 is an electron microscope image of the sound-sensitive agent with the characteristic of independent oxygen in the tumor microenvironment prepared in this example, and as can be seen from fig. 2, the sound-sensitive agent is spherical uniform nano-particles, and the particle size is about 5 nm.

Fig. 3 is a graph showing the particle size distribution of the sound-sensitive agent prepared in this example, which is independent of oxygen in the tumor microenvironment, and as can be seen from fig. 3, the particle size distribution of the sound-sensitive agent is 5.29±0.03 nm, which is substantially consistent with the result of the electron microscope image of fig. 2.

FIG. 4 is a graph showing the ultraviolet absorption spectrum of the sound-sensitive agent with the characteristic independent of oxygen in the tumor microenvironment prepared in this example, and as can be seen from FIG. 4, the sound-sensitive agent has an absorption peak around 290 and nm.

FIG. 5 is a solid electron paramagnetic resonance spectrum of the sound-sensitive agent with the characteristic independent of oxygen in the tumor microenvironment prepared in the embodiment, and as can be seen from FIG. 5, the sound-sensitive agent has a rich oxygen vacancy structure.

And (3) taking TEMP as a single-wire oxygen capturing agent, and detecting single-wire oxygen generated in acoustic power by utilizing electron spin resonance. The ultrasonic parameters used were 40 kHz, 3.0. 3.0W/cm ² 50% duty cycle, total irradiation time was 2 minutes. Fig. 6 shows that the sonosensitizer prepared in this example has stronger electron spin resonance peak under the ultrasonic condition compared with pure water, and the sonosensitizer prepared in this example has the characteristic of independent oxygen in the tumor microenvironment, and can generate abundant singlet oxygen under the ultrasonic radiation.

Mouse breast cancer 4T1 cells (1 x 10 ⁴ And/well) was incubated in 96-well plates for 24 hours, and then cell culture media containing the sonosensitizer prepared in this example and having oxygen independent properties in the tumor microenvironment was added thereto, and incubation was continued under normoxic and hypoxic conditions for 4 hours, thereby examining the sonodynamic therapeutic effect. Experimental facilityControl group 1 (sonosensitizer alone), control group 2 (sonicator alone) and experimental group 3 (sonosensitizer+sonicator) were counted. The parameter of the ultrasound was 40 kHz, 3.0W/cm ² The viability of the cells was examined using the WST-1 method with a 50% duty cycle and an ultrasound irradiation time of 5 minutes. FIG. 7 is a bar graph showing the effect of sonosensitizers with oxygen independent properties in tumor microenvironment on cancer cell survival after ultrasound under normoxic and hypoxic conditions in this example. Experimental results show that the sound-power effect of the sound-sensitive agent, the inhibition effect of the hypoxia condition on cancer cells is similar to that of the normoxic condition, and the inhibition effect can reach more than 85%.

The sonosensitizer with the characteristic of independent oxygen in the tumor microenvironment prepared in the embodiment is injected into a tumor-bearing mouse by tail vein injection, and the sonodynamic treatment is respectively carried out on the first day and the second day, wherein the ultrasonic parameters are 40 kHz and 3.0W/cm ² 50% duty cycle, ultrasound for 5 minutes. Experimental design control group 1 (no treatment), control group 2 (sonosensitizer alone), control group 3 (ultrasound alone) and experimental group 4 (sonosensitizer+ultrasound). The inventors also tested the volume change of each group of mouse tumors during sonodynamic therapy. The experimental results show that compared with other control groups, the tumor growth of the mice in the sound sensitizer and ultrasonic group is obviously inhibited.

Example 2

(2) Continuously heating the system to 260 ℃, adding 0.25 millimole of copper acetylacetonate, and reacting for 30 minutes to obtain copper doped manganese oxide nano particles;

(3) Finally precipitating the copper doped manganese oxide nano particles by ethanol, centrifuging, and modifying methoxy-polyethylene glycol-phospholipid on the surface to change the surface from hydrophobic to hydrophilic.

Through tests, the prepared sound-sensitive agent prepared in the embodiment has the characteristics independent of oxygen in the tumor microenvironment, such as an electron microscope image, a particle size distribution diagram, an ultraviolet absorption spectrum image, a solid electron paramagnetic resonance spectrum image, a histogram for influencing the survival rate of cancer cells, and a sound power treatment effect, which are similar to those in the embodiment 1.

Example 3

(1) Manganese acetylacetonate 0.5 mmol, oleic acid 1 ml, oleylamine 0.5 ml and 1-octadecene 20 ml are mixed, heated to 70 ℃ and kept for reaction for 3 hours;

(2) Continuously heating the system to 320 ℃, adding 0.025 mmol of ferric acetylacetonate, and reacting for 30 minutes to obtain iron-doped manganese oxide nano particles;

(3) Finally precipitating iron doped manganese oxide nano particles by ethanol, centrifuging, and modifying methoxy-polyethylene glycol-phospholipid on the surface to change the surface from hydrophobic to hydrophilic.

The test shows that the particle size of the sound-sensitive agent with the characteristic of independent oxygen in the tumor microenvironment is larger than the particle size synthesized in the benzyl ether solvent at 260 ℃. The ultraviolet absorption spectrum, solid electron paramagnetic resonance spectrum, histogram of the effect on the survival rate of cancer cells, and the effect of photodynamic therapy are similar to those of example 1.

Example 4

(1) Manganese acetylacetonate 0.5 mmol, oleic acid 0.5 ml, oleylamine 1 ml and dibenzyl ether 20 ml were mixed and heated to 70 ℃ and kept for reaction for 4 hours;

(2) Continuously heating the system to 280 ℃, adding 0.05 millimole of ferric acetylacetonate, and reacting for 30 minutes to obtain iron-doped manganese oxide nano particles;

The particle size of the sound-sensitive agent with the characteristic independent of oxygen in the tumor microenvironment prepared by the method is not equal to that of particles synthesized in a benzyl ether solvent at 260 ℃ through testing. The ultraviolet absorption spectrum, solid electron paramagnetic resonance spectrum, histogram of the effect on the survival rate of cancer cells, and the effect of photodynamic therapy are similar to those of example 1.

Example 5

(1) Manganese acetylacetonate 0.5 mmol, oleic acid 1 ml, oleylamine 1 ml and dibenzyl ether 20 ml are mixed, heated to 90 ℃ and kept for reaction for 2 hours;

(2) Continuously heating the system to 260 ℃, adding 0.1 millimole of ferric acetylacetonate, and reacting for 60 minutes to obtain iron-doped manganese oxide nano particles;

(3) Finally, precipitating iron doped manganese oxide nano particles by using acetone, centrifuging, and modifying polyethylene glycol grafted polymaleic anhydride-1-octadecene on the surface to change the surface from hydrophobic to hydrophilic.

Through tests, the prepared sound-sensitive agent prepared in the embodiment has the characteristics independent of oxygen in the tumor microenvironment, such as an electron microscope image, a particle size distribution diagram, an ultraviolet absorption spectrum image, a solid electron paramagnetic resonance spectrum image, a histogram for influencing the survival rate of cancer cells, and a sound power treatment effect, which are similar to those in the embodiment 4.

In addition, the inventors have conducted experiments with other materials, process operations, and process conditions as described in this specification with reference to the foregoing examples, and have all obtained desirable results.

While the invention has been described with reference to an illustrative embodiment, it will be understood by those skilled in the art that various other changes, omissions and/or additions may be made and substantial equivalents may be substituted for elements thereof without departing from the spirit and scope of the invention. In addition, many modifications may be made to adapt a particular situation or material to the teachings of the invention without departing from the scope thereof. Therefore, it is intended that the invention not be limited to the particular embodiment disclosed for carrying out this invention, but that the invention will include all embodiments falling within the scope of the appended claims.

Claims

1. The preparation method of the inorganic sound sensitizer independent of oxygen in tumor microenvironment is characterized by comprising the following steps:

heating the obtained reaction system to 260-320 ℃ in a protective atmosphere, adding a second precursor to form a second mixed reaction system, and continuing the second reaction for 15-60 min to obtain metal doped manganese oxide nano particles, wherein the second precursor is ferric acetylacetonate;

and modifying amphiphilic polyethylene glycol on the surface of the metal-doped manganese oxide nanoparticle to obtain the inorganic sound sensitizer which does not depend on oxygen in the tumor microenvironment, wherein the amphiphilic polyethylene glycol is methoxy-polyethylene glycol-phospholipid.

2. The method of manufacturing according to claim 1, characterized in that: the molar volume ratio of the first precursor, the oleic acid and the oleylamine is 0.5-1 mmol, 0.5-1 mL and 0.5-1 mL.

3. The method of manufacturing according to claim 1, characterized in that: the high boiling point organic solvent is at least one selected from benzyl ether and 1-octadecene.

4. The method of manufacturing according to claim 1, comprising: and heating the first mixed reaction system to 260-320 ℃, wherein the mol ratio of the added second precursor to the first precursor is 5-25:100.

5. The method of manufacturing according to claim 1, characterized in that: the mass ratio of the amphiphilic polyethylene glycol to the whole inorganic sound sensitizer is 5:1-10:1.

6. The method of manufacturing according to claim 1, comprising: after the second reaction is completed, metal doped manganese oxide nanoparticles are precipitated with a polar solvent selected from acetone and/or ethanol, followed by centrifugation.

7. The method of manufacturing according to claim 6, wherein: the volume ratio of the polar solvent to the second mixed reaction system is greater than 10:1.

8. The inorganic sound-sensitive agent independent of oxygen in a tumor microenvironment prepared by the method of any one of claims 1-7, wherein the inorganic sound-sensitive agent independent of oxygen in the tumor microenvironment is manganese oxide nano particles with Fe/Mn metal doping mole ratio of 6.7% -20%.

9. The inorganic sound-sensitive agent independent of oxygen in tumor microenvironment according to claim 8, wherein: the particle size of the inorganic sound sensitizer independent of oxygen in the tumor microenvironment is 5-20 nm.

10. The inorganic sound-sensitive agent independent of oxygen in tumor microenvironment according to claim 8, wherein: the inorganic sound sensitizer independent of oxygen in the tumor microenvironment has uniform particle size and particle size below 10 nm.

11. Use of an inorganic sound-sensitive agent according to any one of claims 8-10 independent of oxygen in a tumor microenvironment for the preparation of an antibacterial product or a product for the photodynamic treatment of tumors.