阅读说明：本技术 金诱导的聚吡咯/二氧化锰纳米材料及其制备方法与应用 (Gold-induced polypyrrole/manganese dioxide nano material and preparation method and application thereof ) 是由 梁华震 林华明 黄其文 黎昌国 于 2021-08-10 设计创作，主要内容包括：本发明属于生物医药技术领域,具体涉及金诱导的聚吡咯/二氧化锰纳米材料及其制备方法与应用。首次将含金氧化剂诱导引发氧化聚合制备聚吡咯,制备方法简单且粒径可控；所得材料生物相容性好,并且同时具有很好的放射增敏性、光热效应,并且可以显著改善肿瘤微环境,到达肿瘤部位后可以协同增效达到显著杀死肿瘤细胞的效果,并且对机体损伤程度较小,安全性高,具有较好的药物研发前景。(The invention belongs to the technical field of biological medicines, and particularly relates to a gold-induced polypyrrole/manganese dioxide nano material as well as a preparation method and application thereof. The gold-containing oxidant is induced to initiate oxidative polymerization to prepare polypyrrole for the first time, and the preparation method is simple and controllable in particle size; the obtained material has good biocompatibility, good radiosensitization and photothermal effects, can obviously improve the tumor microenvironment, can achieve the effect of obviously killing tumor cells by synergy after reaching the tumor part, has small damage degree to the organism and high safety, and has good drug research and development prospect.)

1. A preparation method of gold-induced polypyrrole/manganese dioxide nano material is characterized by comprising the following steps:

s1, mixing the stabilizer, the oxidant and the pyrrole monomer, stirring and reacting at 4-40 ℃, centrifuging, and collecting precipitates to obtain the polypyrrole nano material;

s2, preparing the polypyrrole nano material obtained in the step S1 into a polypyrrole aqueous solution, adding potassium permanganate, reacting completely at 20-60 ℃, centrifuging, collecting precipitate, and performing post-treatment to obtain the polypyrrole nano material;

wherein the oxidant is selected from one or more of tetrachloroauric acid trihydrate, tetrachloroauric acid dihydrate, sodium tetrachloroauric acid and potassium tetrachloroauric acid.

2. The method according to claim 1, wherein in step S1, the stabilizer is one or more selected from polyvinyl alcohol, polyvinylpyrrolidone, and sodium dodecyl sulfate.

3. The method according to claim 1, wherein the stabilizer is added in an amount of 0.1 to 10mg/mL in step S1.

4. The method according to claim 1, wherein in step S1, the oxidizing agent is added after being dissolved in 0.01-30 mM of the oxidizing agent solution in an amount of 1-10 mL.

5. The method according to claim 1, wherein in step S1, the volume ratio of the pyrrole monomer to the total volume of the solution is 1: (10-200).

6. The preparation method according to claim 1, wherein in step S2, the concentration ratio of the potassium permanganate to the polypyrrole nano-materials in the aqueous solution is 1 (0.5-3).

7. A gold-induced polypyrrole/manganese dioxide nanomaterial prepared by the preparation method of any one of claims 1 to 6.

8. The nanomaterial of claim 7, wherein the gold-induced polypyrrole/manganese dioxide nanomaterial has a particle size of 5-200 nm.

9. The use of the gold-induced polypyrrole/manganese dioxide nanomaterial of claim 7 in the preparation of a medicament for the prevention and treatment of tumors.

10. Use according to claim 9, wherein the medicament is combined with radiotherapy and/or photothermal therapy.

Technical Field

The invention belongs to the technical field of biological medicines. More particularly, relates to a gold-induced polypyrrole/manganese dioxide nano material, and a preparation method and application thereof.

Background

Cancer is one of the diseases with the highest fatality rate worldwide, and the number of patients rapidly rises every year, which is a public health problem seriously threatening the survival of human beings. The traditional method for treating cancer mainly comprises surgery, radiotherapy and chemotherapy at present; the radiotherapy (radiotherapy) is a local non-invasive tumor treatment strategy widely used clinically, and uses alpha, beta and gamma ray beams and electron beams to directly damage DNA molecules of tumor cells or generate free radicals to induce apoptosis so as to kill cancer cells. However, single radiotherapy generally requires higher radiation intensity to achieve clinically desirable therapeutic effects, and the toxic and side effects on surrounding normal tissues are increased, which greatly affects the quality of life of patients; in addition, the tumor microenvironment is a severe hypoxic environment, thereby greatly reducing the effect of oxygen-dependent radiotherapy.

In recent years, researchers find that nano materials containing high atomic number metal elements (such as gold, tungsten, bismuth and the like) can be used as radiosensitizers, and the radiotherapy efficiency is improved by enhancing the deposition of ray energy in tumor tissues. For example, the chinese patent application CN108356279A discloses a method for preparing a hollow gold nano material, the prepared hollow gold nano material can be directly loaded with drug molecules, and the effect of treating tumors can be achieved by combining methods such as drug chemotherapy and the like while radiosensitization. However, the problem that the tumor microenvironment hypoxia condition affects the radiotherapy effect still exists, and the conditions of radiation enhancement and toxic and side effects increase are caused. Therefore, a novel medicine which has the effect of sensitizing radiotherapy and can supply oxygen is urgently needed clinically to solve the problems faced by the existing radiotherapy.

Disclosure of Invention

The invention aims to solve the technical problems that the radiation enhancement of the existing tumor radiotherapy can damage normal cells to different degrees, and the improvement of the radiotherapy effect by utilizing a radiosensitizer is limited, and provides a gold-induced polypyrrole/manganese dioxide nano material which can obviously enhance the radiotherapy effect and improve the anti-tumor effect.

The invention aims to provide a preparation method of a polypyrrole/manganese dioxide nano material.

The invention also aims to provide the application of the gold-induced polypyrrole/manganese dioxide nano material in preparing a medicament for preventing and treating tumors.

The above purpose of the invention is realized by the following technical scheme:

a preparation method of gold-induced polypyrrole/manganese dioxide nano material comprises the following steps:

s1, mixing the stabilizer, the oxidant and the pyrrole monomer, stirring and reacting at 4-40 ℃, centrifuging, and collecting precipitates to obtain the polypyrrole nano material;

s2, preparing the polypyrrole nano material obtained in the step S1 into a polypyrrole aqueous solution, adding potassium permanganate, reacting completely at 20-60 ℃, centrifuging, collecting precipitate, and performing post-treatment to obtain the polypyrrole nano material;

wherein the oxidant is selected from one or more of tetrachloroauric acid trihydrate, tetrachloroauric acid dihydrate, sodium tetrachloroauric acid and potassium tetrachloroauric acid.

Further, in step S1, the stabilizer is selected from one or more of polyvinyl alcohol, polyvinylpyrrolidone, and sodium lauryl sulfate.

Further, in step S1, the amount of the stabilizer added is 0.1 to 10 mg/mL.

Further, in step S1, the oxidant is added after being prepared into a solution, the concentration of the oxidant solution is 0.01-30 mM, the addition amount is 1-10 mL, and the solvent is water.

Further, in step S1, the volume ratio of the pyrrole monomer to the total volume of the solution is 1: (10-200).

Preferably, in step S1, the stirring reaction time is 1 to 60 minutes, and the stirring rotation speed is 200 to 1500 rpm.

Further, in step S2, the concentration ratio of the potassium permanganate to the polypyrrole nano-material in the aqueous solution is 1 (0.5-3).

Preferably, in step S2, the reaction time is 0.5 to 2 hours, and stirring may be performed during the reaction, wherein the stirring speed is 100 to 1500 rpm.

Preferably, in the steps S1 and S2, the rotation speed of the centrifugation is 5000-30000 rpm, and the centrifugation time is 5-30 min.

In addition, the invention also provides a gold-induced polypyrrole/manganese dioxide nano material prepared by the preparation method.

Further, the particle size of the gold-induced polypyrrole/manganese dioxide nano material is 5-200 nm.

Still further, the gold-induced polypyrrole/manganese dioxide nanomaterial has a dodecahedral, cubic, spherical, rod-like, or irregular polyhedral structure.

In addition, the invention also provides application of the gold-induced polypyrrole/manganese dioxide nano material in preparing a medicine for preventing and treating tumors.

Preferably, the medicine can be combined with photothermal therapy and radiation therapy when treating tumors, so that better tumor removal and treatment effects can be achieved.

The gold-induced polypyrrole/manganese dioxide nano material provided by the invention is induced by a gold-containing oxidant for the first time to initiate oxidative polymerization to prepare polypyrrole, so that a gold element exists in the prepared nano material, the gold-induced polypyrrole/manganese dioxide nano material has a radiosensitization effect, and the radiotherapy effect can be obviously improved; the manganese dioxide is coated on the periphery of the material, so that the oxygen concentration of a tumor microenvironment can be obviously improved under an anoxic tumor specific microenvironment, the anti-tumor effect of an oxygen-dependent radiotherapy method is further enhanced, the radiation intensity required by radiotherapy is synergistically reduced from the two aspects, and the damage of radiotherapy to organism tissues is obviously reduced. In addition, the gold-induced polypyrrole/manganese dioxide nano material provided by the invention has a photo-thermal property, and can exert a strong photo-thermal effect to cauterize tumors under the laser stimulation of a near-infrared region II. After the gold-induced polypyrrole/manganese dioxide nano material provided by the invention is further targeted to a tumor part, the gold-induced polypyrrole/manganese dioxide nano material is cooperated with a photothermal therapy and/or a radiotherapy, so that a better tumor treatment effect can be achieved.

The invention has the following beneficial effects:

the invention provides a gold-induced polypyrrole/manganese dioxide nano material, which is prepared by inducing an oxidizing agent containing gold to initiate oxidative polymerization for the first time, and has the advantages of simple preparation method and controllable particle size; the obtained material has good biocompatibility, good radiosensitization and photothermal effects, can obviously improve the tumor microenvironment, can achieve the effect of obviously killing tumor cells by synergy after reaching the tumor part, has small damage degree to the organism and high safety, and has good drug research and development prospect.

Drawings

FIG. 1 is an electron microscope scanning image of AuPPy nano-material obtained in example 1 of the present invention.

FIG. 2 shows AuPPy @ MnO obtained in example 1 of the present invention₂Electron microscopy scan of nanocomposite.

FIG. 3 shows AuPPy @ MnO obtained in example 1 of the present invention₂Energy dispersive X-ray spectroscopy of the nanocomposites.

FIG. 4 shows AuPPy nanomaterial and AuPPy @ MnO obtained in example 1 of the present invention₂X-ray photoelectron spectrum of the nanocomposite.

FIG. 5 shows AuPPy nanomaterial and AuPPy @ MnO obtained in example 1 of the present invention₂Particle size distribution profile of the nanocomposite.

FIG. 6 shows AuPPy nanomaterial and AuPPy @ MnO obtained in example 1 of the present invention₂Zeta potential profile of the nanocomposite.

FIG. 7 shows AuPPy @ MnO obtained in example 1 of the present invention₂Ultraviolet-visible spectrum of the nanocomposite.

FIG. 8 shows AuPPy @ MnO obtained in example 1 of the present invention₂A photo-thermal performance evaluation graph of the nanocomposite; wherein, A-1064nm laser excitation, B-photo-thermal stability evaluation chart.

FIG. 9 shows AuPPy @ MnO obtained in example 1 of the present invention₂Statistical map of the oxygen production capacity of the nanocomposite.

FIG. 10 shows AuPPy @ MnO obtained in example 1 of the present invention₂Glutathione consumption capacity study statistical chart of the nano composite material.

Detailed Description

The invention is further described with reference to the drawings and the following detailed description, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.

Unless otherwise indicated, reagents and materials used in the following examples are commercially available.

Example 1 gold-induced polypyrrole/manganese dioxide nanomaterial

The preparation method of the gold-induced polypyrrole/manganese dioxide nano material comprises the following steps:

s1 preparation of gold-induced polymerized polypyrrole nano material (AuPPy)

Adding 10mL of deionized water into a 50mL single-neck round-bottom flask, adding 50mg of polyvinyl alcohol, mildly stirring at a heating environment of 90 ℃, condensing and refluxing for 1 hour, and naturally cooling to room temperature; dropwise adding 5mL of tetrachloroauric acid trihydrate aqueous solution (10mM) under the stirring condition of the rotation speed of 500-1200 rpm, and continuously stirring for 1 hour; dropwise adding 200 mu L of pyrrole monomer, and stirring at 500-1200 rpm for reaction for 15 minutes; centrifuging the reaction solution for 20min by using a low-temperature ultracentrifuge at 20000rpm/min, and collecting the precipitate to obtain a gold-induced polymerization polypyrrole AuPPy nano material;

s2, gold-induced polymerization polypyrrole/manganese dioxide nanocomposite (AuPPy @ MnO)₂) Preparation of

Dropwise adding 25mL of potassium permanganate aqueous solution (200 mu g/mL) into 25mL of AuPPy aqueous solution (200 mu g/mL) prepared from the AuPPy nano material obtained in the step S1 under the condition of low-speed stirring at 200rpm, transferring the AuPPy aqueous solution into a constant-temperature water bath kettle at 40 ℃, and carrying out vigorous stirring reaction at 1000rpm for 1 hour; centrifuging at 15000rpm/min for 10min after the reaction is finished, collecting precipitate, washing with deionized water for three times to obtain the gold-induced polymerized polypyrrole/manganese dioxide nanocomposite (AuPPy @ MnO)₂)。

Example 2 gold-induced polypyrrole/manganese dioxide nanomaterial

The preparation method of the gold-induced polypyrrole/manganese dioxide nano material comprises the following steps:

s1 preparation of gold-induced polymerized polypyrrole nano material (AuPPy)

Adding 10mL of deionized water into a 50mL single-neck round-bottom flask, adding 50mg of polyvinylpyrrolidone, mildly stirring at a heating environment of 90 ℃, condensing and refluxing for 1 hour, and naturally cooling to room temperature; dropwise adding 5mL of tetrachloroauric acid dihydrate aqueous solution (10mM) under the stirring condition of the rotation speed of 500-1200 rpm, and continuing stirring for 1 hour; dropwise adding 200 mu L of pyrrole monomer, and stirring at 500-1200 rpm for reaction for 15 minutes; centrifuging the reaction solution for 10min at 30000rpm/min by using a low-temperature ultracentrifuge, and collecting precipitates to obtain a gold-induced polymerization polypyrrole AuPPy nano material;

s2, gold-induced polymerization polypyrrole/manganese dioxide nanocomposite (AuPPy @ MnO)₂) Preparation of

Dropwise adding 25mL of potassium permanganate aqueous solution (200 mu g/mL) into 25mL of AuPPy aqueous solution (200 mu g/mL) prepared from the AuPPy nano material obtained in the step S1 under the condition of low-speed stirring at 200rpm, transferring the AuPPy aqueous solution into a constant-temperature water bath kettle at 50 ℃, and carrying out vigorous stirring reaction at 1000rpm for 1 hour; centrifuging at 20000rpm/min for 10min after the reaction is finished, collecting precipitate, washing with deionized water for three times to obtain gold-induced polymerization polypyrrole/manganese dioxide nanocomposite (AuPPy @ MnO)₂)。

The gold-induced polypyrrole/manganese dioxide nanomaterial prepared in example 1 is used as an example to perform structural characteristics and performance tests, and other examples can achieve similar effects.

Test example 1 determination of physical and chemical Properties of gold-induced polypyrrole/manganese dioxide nanomaterial

The polypyrrole AuPPy nanomaterial obtained by gold induced polymerization in step S1 of example 1 was subjected to electron microscope scanning analysis, and the result is shown in fig. 1. As can be seen from the figure, the polypyrrole AuPPy nanometer material obtained by the gold-induced polymerization in the example 1 shows uniform loading of ultra-small gold nanoparticles, and the AuPPy nanometer material has uniform particle size distribution and the size of 35-50 nm.

For AuPPy @ MnO obtained in step S2 of example 1₂The nanocomposite was subjected to electron microscopy scanning analysis and the results are shown in figure 2. As can be seen, AuPPy @ MnO obtained in example 1₂The surface of the nano composite material becomes rough and the size is largerThe size of the AuPPy nano material is increased to 40-55 nm, which indicates that the manganese dioxide is successfully coated.

For AuPPy @ MnO obtained in example 1₂The nanocomposite was subjected to energy dispersive X-ray spectroscopy, and the results are shown in fig. 3. As can be seen, AuPPy @ MnO₂The existence of Mn, Au, C, N and O elements in the nano composite material indicates that the AuPPy @ MnO of the invention₂The nano composite material is successfully prepared.

AuPPy nanomaterial obtained in example 1, AuPPy @ MnO₂The X-ray photoelectron spectroscopy (XPS) analysis of the nanocomposite material resulted in the results shown in FIG. 4. As can be seen, AuPPy @ MnO is relative to the XPS spectrum of AuPPy nano material₂Mn element is newly added in the nano composite material, which shows the successful coating of manganese dioxide and AuPPy @ MnO₂Successful preparation of nanocomposites.

AuPPy nanomaterial obtained in example 1, AuPPy @ MnO₂The results of the particle size distribution analysis of the nanocomposites are shown in FIG. 5. As can be seen, the hydrodynamic particle size of the AuPPy nano material is about 134nm, and the AuPPy @ MnO is₂The hydrodynamic particle size of the nanocomposite was increased, at around 186nm, indicating successful coating of manganese dioxide in the present invention.

AuPPy nanomaterial obtained in example 1, AuPPy @ MnO₂The results of Zeta potential analysis of each nanocomposite are shown in FIG. 6. As can be seen from the figure, the Zeta potential of the AuPPy nano material is about 8.32mV, and the AuPPy @ MnO is coated by manganese dioxide₂The Zeta potential of the nano composite material is about-33.97 mV.

Experimental example 2AuPPy @ MnO₂Determination of optical Properties of nanocomposites

For AuPPy @ MnO obtained in example 1₂The optical absorption intensity of the nanocomposite was measured, and the results are shown in fig. 7. As can be seen, AuPPy @ MnO₂The nano composite material has strong optical absorption in ultraviolet-visible light-near infrared region and has concentration dependence.

Under the condition of laser with wavelength of 1064nm, AuPPy @ MnO with different concentrations₂NanocompositeThe photothermal properties of the materials were evaluated and the results are shown in fig. 8.

As can be seen in FIG. 8A, AuPPy @ MnO₂The nano composite material has higher temperature rise under the excitation of 1064nm laser and excellent photo-thermal performance. As can be seen in FIG. 8B, AuPPy @ MnO₂After the nano composite material is irradiated by five cycles of 1064nm laser, the nano composite material still shows strong light stability, and the phenomenon of weakened photo-thermal performance does not occur. To sum up, AuPPy @ MnO₂The nano composite material has the temperature required by photo-thermal treatment for burning tumors.

Experimental example 3AuPPy @ MnO₂Determination of tumor microenvironment improvement performance of nano composite material

For AuPPy @ MnO obtained in example 1₂The oxygen production capacity of the nano composite material is tested, and the testing method comprises the following steps: different concentrations of AuPPy @ MnO₂The nanocomposite was dispersed in phosphate buffer (pH 7.4) and H was added once with a syringe₂O₂(10mM), the oxygen concentration of the solution at different time points was measured using a JPSJ-605F portable dissolved oxygen meter. The results are shown in FIG. 9.

As can be seen, AuPPy @ MnO₂The nano composite material has strong oxygen generation capacity and concentration dependence, and can relieve the tumor hypoxia state and promote radiotherapy.

For AuPPy @ MnO obtained in example 1₂The glutathione consumption capability of the nano composite material is tested, and the test method comprises the following steps: different concentrations of AuPPy @ MnO₂The nanocomposite was dispersed in phosphate buffer (pH 7.4), then 0.1mM GSH was added, and finally the consumption of GSH was detected using a 5,5' -dithiobis (2-nitrobenzoic acid) probe. The results are shown in FIG. 10.

As can be seen, AuPPy @ MnO₂The nano composite material has obvious glutathione removing capacity, can reduce the oxidation resistance of tumors and further promotes the radiotherapy effect.

To sum up, AuPPy @ MnO₂The nano composite material has the capabilities of improving the oxygen concentration of a tumor microenvironment and consuming glutathione, can assist in enhancing the radiotherapy effect, and has the capabilities of killing tumors and preventing tumor recurrence and metastasis.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.