CN113117077B - Platinum-based monatomic nanoenzyme for tumor combined treatment and preparation method thereof - Google Patents
Platinum-based monatomic nanoenzyme for tumor combined treatment and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a platinum-based monatomic nanoenzyme for tumor combined treatment and a preparation method thereof, and a platinum (IV) -based monatomic nanoenzyme medicine is prepared by taking cisplatin and 3, 4-diaminopyridine as precursors through oxidative polymerization. Compared with the prior art, the platinum (IV) group monatomic nanoenzyme medicine prepared by the invention has catalase-like and peroxidase-like activities, can greatly relieve tumor hypoxia, consumes glutathione overexpressed in tumors, can promote generation of free radicals in tumor cells, reduces toxic and side effects, and improves radiotherapy treatment effects. The nano enzyme has strong absorption in the whole near infrared region, can absorb 808nm laser and convert the laser into high-energy heat when being used as a photo-thermal conversion agent, greatly improves the killing power to tumor cells, and realizes photo-thermal treatment, so that the thermal therapy and radiotherapy functions of the platinum (IV) group monoatomic nano enzyme medicament have strong killing effect on the tumor cells.
Description
Technical Field
The invention belongs to the technical field of nano materials, and particularly relates to platinum-based monatomic nanoenzyme for tumor combined treatment and a preparation method thereof.
Background
Cancer, a disease with high mortality rate and difficult cure, always causes extreme troubles to people's lives. From the data of the world cancer report in 2015, the worldwide cancer cases are rapidly increased all the time, the occurrence of cancers in China has a trend of being younger, higher in incidence rate and mortality rate by three lines in recent 20 years, and the number of cases of cancer death in China is up to 270 ten thousand every year. At present, the treatment methods for treating cancers mainly comprise surgical treatment, radiotherapy and chemotherapy. However, these treatments have great side effects and are prone to adverse consequences, which makes it difficult to meet clinical needs. Therefore, in order to effectively treat cancer, people begin to search for some nano-drugs with simple synthetic method, high biological safety and good tumor cell inhibition effect, and cooperate with common treatment means to treat cancer
The photothermal therapy (PTT) is a therapeutic method in which a material having a high photothermal conversion efficiency is injected into the inside of a human body, is concentrated near tumor tissue by a targeting recognition technology, and converts light energy into heat energy under the irradiation of an external light source (generally, near infrared light) to kill cancer cells. Photothermal therapy has good tissue penetration and the photothermal material itself has no significant toxicity. Secondly, the photothermal therapy has high specificity and good targeting property, can selectively kill and kill tumor cells, avoids unnecessary damage to normal tissues, and reduces toxic and side effects.
Radiotherapy (RT), one of the most widespread methods for treating cancer in clinic, is based on the principle that a large amount of cytotoxic Reactive Oxygen Species (ROS) is generated by the interaction of X-rays and ionizing radiation with water or oxygen inside tissues to induce DNA double strand damage in cells. Radiation therapy can invisibly limit the DNA that induces oxidative stress and destroys tumor cells, but has little effect on hypoxic tumor cells that are resistant to radiation. In the radiation treatment process, the ion-doped polypyridine nano-drug is used as an anti-tumor nano-drug, the excellent oxidoreductase activity of the ion-doped polypyridine nano-drug is utilized to regulate and control a tumor microenvironment, the excellent anti-tumor effect can be realized, and the polypyridine nano-drug (M-PDAP) has good biocompatibility, high specific surface area, doped ions capable of generating charge transfer and high photo-thermal conversion efficiency.
How to utilize the multifunctionality of the nano particles, the activity of peroxidase mimic of the nano enzyme is expanded, and the nano enzyme is organically combined with traditional anticancer means such as chemotherapy, radiotherapy, surgical treatment and the like, so that the anticancer efficiency is improved, and the multifunctional anticancer application of the nano enzyme is realized. Therefore, the nano enzyme system with the tumor microenvironment regulation function is designed, and the immunosuppression tumor microenvironment can be reversed, so that the synergistic anticancer treatment is realized.
In the prior art, a bivalent platinum (II) group and 3, 4-diaminopyridine are used as precursors to prepare a novel platinum (II) -based polypyridine nano-drug (Pt (II) -PDAP), the effect of the novel platinum (II) -based polypyridine nano-drug is mainly that the novel nano-drug has strong absorption in a near infrared region, and the nano-drug is monitored by an infrared thermal imager to have good photo-thermal conversion efficiency and light stability, so that the platinum (II) -based polypyridine nano-drug (Pt (II) -PDAP) can be used as a photo-thermal absorption reagent for bacterial photo-thermal treatment, but only has a single photo-thermal effect and can not realize a radiotherapy effect with double enzyme activity.
Disclosure of Invention
In view of the above, the present invention provides a platinum-based monatomic nanoenzyme for tumor combination therapy and a preparation method thereof, so that the platinum (iv) -based monatomic nanoenzyme has both catalase-like and peroxidase-like activities, can greatly alleviate tumor hypoxia, consume over-expressed glutathione in tumors, can promote the generation of free radicals in tumor cells, reduce toxic and side effects, and improve the therapeutic effect of radiotherapy. The nano enzyme has strong absorption in the whole near infrared region, can absorb 808nm laser and convert the laser into high-energy heat when being used as a photo-thermal conversion agent, greatly improves the killing power to tumor cells, and realizes photo-thermal treatment, so that the thermal therapy and radiotherapy functions of the platinum (IV) group monoatomic nano enzyme medicament have strong killing effect on the tumor cells.
In order to achieve the purpose, the invention adopts the following technical scheme:
a platinum-based monatomic nanoenzyme, which is composed of cisplatin cis- [ PtCl [ ] 2 (NH 3 ) 2 ]The hydrogen peroxide, the 3, 4-diaminopyridine and the ultrapure water are prepared by oxidative polymerization, and the 3, 4-diaminopyridine is a monomolecular crystal.
Furthermore, the particle size of the nano enzyme is 5nm-200nm, and the nano enzyme particles are in a fusiform irregular polyhedral structure.
A preparation method of platinum-based monatomic nanoenzyme is characterized by comprising the following steps:
s1, adding cisplatin and hydrogen peroxide into ultrapure water, stirring to obtain a solution I, and sequentially recrystallizing, filtering, washing and drying the solution I to obtain powder A;
s2, dissolving the powder A in hydrochloric acid, and stirring to obtain a solution II;
s3, adding ultrapure water into the solution II, then adding 3, 4-diaminopyridine, ultrasonically dissolving, and stirring for reacting to obtain a solution III;
s4, dialyzing and purifying the solution III, and then freeze-drying to obtain a solid sample material;
and S5, performing index detection on the solid sample obtained in the step S4.
Further, in the step S1, the molar ratio of cisplatin, hydrogen peroxide, and ultrapure water is 1: 102: 417; the stirring speed is 600-800 r/min, the stirring temperature is 50-60 ℃, and the stirring time is 1-1.2 h.
Further, in the step S1, the molar ratio of cisplatin, hydrogen peroxide, and ultrapure water is 1: 105:420.
Further, in step S2, the stirring manner is: firstly stirring at the rotating speed of 400-600 r/min for 1-1.5h at the temperature of 40-60 ℃, and then stirring at the rotating speed of 400-600 r/min for 6-18h at normal temperature.
Further, the concentration of the hydrochloric acid solution in the step S2 is 10 mol/L.
Further, in the step S3, the ultrasonic frequency is 55-75 kHz, the reaction condition is that the rotating speed is 600-800 r/min, the reaction is continuously carried out for 24-26h at the temperature of 37-40 ℃, and the molar ratio of the solution II to the 3, 4-diaminopyridine is 1: 3.
Further, in the step S4, dialysis is performed by using a dialysis bag with a molecular weight of 12.0kDa for dialysis and purification, the dialysate is deionized water, the dialysis temperature is 25-35 ℃, the deionized water is replaced every 2-3 hours, and the process is repeated for 3 times.
Further, the platinum-based monatomic nanoenzyme is applied to the preparation of a synergistic medicine for assisting tumor thermotherapy and radiotherapy functions.
Has the advantages that:
the invention has the beneficial effects that:
(1) the preparation method of the platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) is simple, convenient and quick, has low pollution and good repeatability, and can catalyze hydrogen peroxide (H) under acidic conditions 2 O 2 ) Decompose to OH, catalyze hydrogen peroxide (H) under neutral conditions 2 O 2 ) Decomposition into H 2 O and O 2 In which highly toxic hydroxyl radicals (. OH) are produced, can induce apoptosis or necrosis of tumor cells.
(2) The platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) integrates chemotherapy and nano particles with near-infrared light responsiveness for anti-tumor treatment, has good photothermal conversion capability, excellent water dispersibility and tumor tissue targeting property, and can effectively kill tumor cells.
In summary, the technical scheme of the application is Pt from the chemical point of view 2+ /Pt 4+ The existence of redox couple makes it possess both catalase-like and peroxidase-like activities, Fenton-like reaction makes it exhibit excellent OH generating ability to eliminate GSH over-expressed in TME, and reduces the anti-oxidation ability of tumor by GSH peroxidase-like activity, and combines with endogenous H 2 O 2 Interaction to produce O 2 And relieve hypoxia of TME. From the physical point of view, the Pt (IV) -PDAP and X-ray action process mainly comprises the generation of photoelectric effect, scattering (Compton scattering and Rayleigh scattering) and electron pair effect. The invention adoptsMonitoring a photocurrent signal generated by the nano-drug under X-ray irradiation by using an electrochemical workstation, theoretically judging a transfer path of excited electrons under X-ray irradiation, and further determining the specific type of generated ROS. Meanwhile, the material has strong absorption in a near infrared region, has good photo-thermal conversion capability, can enhance local radiotherapy dosage, combines thermal therapy and radiotherapy, supplements each other, and improves the tumor killing power. The effect of the technical scheme is obviously different from the existing divalent platinum (II) -based polypyridine nano-drug preparation technology, and the synthesized novel nano-material drug has better anti-tumor effect by changing raw materials and a preparation method. The main difference lies in that the compound has double-enzyme activity and a heat radiation treatment effect, has double effects, synergy and better application prospect.
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In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present application, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a transmission electron microscope image of a platinum (IV) based monatomic nanoenzyme (Pt-PDAP) prepared in example 2 of the present invention;
FIG. 2 is an X-ray energy spectrum of a platinum (IV) group monatomic nanoenzyme (Pt-PDAP) prepared in example 2 of the present invention;
FIG. 3 is an infrared spectrum of a platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) prepared in example 2 of the present invention;
FIG. 4 is a UV spectrum of a platinum (IV) based monatomic nanoenzyme (Pt-PDAP) prepared in example 2 of the present invention;
FIG. 5 is a kinetic map of a platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) prepared in example 2 of the present invention;
FIG. 6 is a kinetic map of different hydrogen peroxide concentrations for a platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) prepared in example 2 of the present invention;
FIG. 7 is a graph showing the concentration-dependent change of a platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) as a peroxidase, prepared in example 2 of the present invention;
FIG. 8 is a graph showing the change of catalytic effect of a platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) as a peroxidase, prepared in example 2 of the present invention, under different hydrogen peroxide concentrations;
FIG. 9 is a graph showing the change of catalytic effects of a platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) as a peroxidase at different temperatures, prepared in example 2 of the present invention;
FIG. 10 is a graph showing the change in catalytic effect of a platinum (IV) -based monatin nanoenzyme (Pt-PDAP) as a peroxidase as prepared in example 2 of the present invention at different pHs;
FIG. 11 is a diagram showing a platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) prepared in example 2 of the present invention as a reactive oxygen species;
FIG. 12 is a graph showing the detection of platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) as active oxygen at various concentrations, prepared in example 2 of the present invention;
FIG. 13 is a graph showing the change in the absorption peak of reduced glutathione after the addition of reduced glutathione to a platinum (IV) -based monatin nanoenzyme (Pt-PDAP) prepared in example 2 at various concentrations;
FIG. 14 is a graph showing the change in the absorption peak of reduced glutathione after platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) prepared in example 2 and reduced glutathione were added to PDAP;
FIG. 15 is a graph showing the killing effect of platinum (IV) based monatomic nanoenzyme (Pt-PDAP) in combination with hyperthermia radiation therapy on T4 cells prepared in example 2 of the present invention;
FIG. 16 is a graph showing the tumor-suppressing effect of different treatment modalities after intratumoral administration of a platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) prepared in example 2 of the present invention to tumor-bearing mice. Blank, II NIR, III X-ray, IV X-ray +808nm, V material, VI material + NIR, VII material + X-ray, VIII material + NIR + X-ray, NIR: near infrared light; PBS: PBS buffer.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1: preparation of platinum based monatomic nanoenzyme
(1) 0.2g of cisplatin (cis- [ PtCl) 2 (NH 3 ) 2 ]) And 7mL of 30% hydrogen peroxide (H) 2 O 2 ) Adding into a round-bottom flask with 5mL of ultrapure water, placing the round-bottom flask into a constant-temperature water bath kettle, stirring at the rotation speed of 600r/min at 50 ℃ for 1 hour to obtain a yellow solution, and sequentially recrystallizing, filtering, washing and drying the obtained solution to obtain powder A;
(2) weighing 0.2g of powder A, dissolving the powder A in 5mL of HCl (10M), placing the solution in a constant-temperature water bath kettle, stirring the solution at the rotation speed of 400r/min at 50 ℃ for 1 hour, and then placing the solution at normal temperature and stirring the solution overnight to obtain a yellow-green solution II;
(3) adding 25mL of ultrapure water into the yellow-green solution II, then weighing 20mmol of 3, 4-diaminopyridine (0.13g) and adding, placing the yellow-green solution II into an ultrasonic dispersion instrument, ultrasonically dissolving under the condition that the ultrasonic frequency is 75kHz, then placing the yellow-green solution II into a constant-temperature water bath, and continuously reacting for 26h under the conditions that the rotating speed is 800r/min and the temperature is 40 ℃ to obtain a solution III;
(4) putting the solution III into a dialysis bag with the molecular weight of 12.0kDa for dialysis and purification at 35 ℃, replacing deionized water once every 3 hours, repeating for 3 times, and then freeze-drying to obtain a solid sample material;
(5) and (4) carrying out index detection on the substance obtained in the step (4).
Example 2: preparation of platinum based monatomic nanoenzyme
(1) 0.4g of cisplatin (cis- [ PtCl) 2 (NH 3 ) 2 ]) And 14mL of 30% hydrogen peroxide (H) 2 O 2 ) Adding into a round-bottom flask containing 10mL of ultrapure water, placing the round-bottom flask into a constant-temperature water bath, stirring at 60 deg.C at 800r/min for 1 hr to obtain yellow solution, sequentially adding the yellow solution into the flaskRecrystallizing, filtering, washing and drying to obtain powder A;
(2) weighing 0.2g of powder A, dissolving the powder A in 5mL of HCl (10M), placing the solution in a constant-temperature water bath kettle, stirring the solution at the rotation speed of 600r/min and the temperature of 50 ℃ for 1 hour, and then placing the solution at the normal temperature and stirring the solution overnight to obtain a yellow-green solution II;
(3) adding 25mL of ultrapure water into the yellow-green solution II, then weighing 20mmol of 3, 4-diaminopyridine (0.13g) and adding, placing the yellow-green solution II into an ultrasonic dispersion instrument, ultrasonically dissolving under the condition that the ultrasonic frequency is 55kHz, then placing the yellow-green solution II into a constant-temperature water bath kettle, and continuously reacting for 24 hours under the conditions that the rotating speed is 600r/min and the temperature is 37 ℃ to obtain a solution III;
(4) putting the solution III into a dialysis bag with the molecular weight of 12.0kDa for dialysis and purification at 25 ℃, replacing deionized water once every 2 hours, repeating for 3 times, and then freeze-drying to obtain a solid sample material;
(5) and (4) carrying out index detection on the substance obtained in the step (4).
1. Characterization of particle size
The platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) obtained in example 2 is characterized by using a transmission electron microscope, and the result is shown in figure 1, and it can be seen from figure 1 that the platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) is uniform in shape, shuttle-shaped, has a particle size of between 5nm and 200nm, and has good dispersibility and uniform particle size.
2. X-ray energy spectrum analysis
An X-ray energy spectrum is used for characterizing the platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) obtained in example 2, and the result is shown in figure 2, and the ion-doped polypyridine polymer nanomaterial contains a large amount of elements such as C, Pt, O, Cl, N and the like from figure 2, which indicates that Pt and 3, 4-diaminopyridine are polymerized together.
3. Characterization of the Infrared Spectrum
A platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) obtained in example 2 was characterized by an infrared spectrometer (Thermo FT-IR200), and the results are shown in FIG. 3, 3280cm -1 、1300cm -1 、800cm -1 Is a telescopic shock absorption of-N-H-, 530cm -1 Has an absorption peak of V Pt-N Characteristic absorption ofA peak; 3150cm -1 、1640cm -1 、1410cm -1 And 688cm -1 Is a characteristic absorption peak of the 3, 4-diaminopyridine, and characteristic peaks of cisplatin and the 3, 4-diaminopyridine can be seen to appear in an infrared spectrogram of a platinum (IV) -based monoatomic nanoenzyme (Pt-PDAP).
4. Characterization of the ultraviolet spectrum
A platinum (IV) group monatomic nanoenzyme (Pt-PDAP) obtained in example 2 was characterized by an ultraviolet spectrometer (UV-Vis), and the results are shown in FIG. 4, in FIG. 4: the nanoparticles do not show a distinct absorption peak in the near infrared region.
5. Evaluation of enzyme kinetic mechanism detection effect
The specific detection process comprises the following steps: peroxidase and H 2 O 2 The reaction can change colorless TMB into blue oxTMB, and obvious absorption peaks can be observed at 370nm and 652 nm. Therefore, the peroxidase-like activity of a platinum (IV) group monatomic nanoenzyme (Pt-PDAP) can be detected by using TMB as a substrate. One platinum (iv) -based monatin nanoenzyme (Pt-PDAP) prepared in example 2 was set at a concentration of 100 μ g/mL, hydrogen peroxide was set at a concentration of 50mM, and 3,3',5,5' -Tetramethylbenzidine (TMB) was set at a gradient concentration, using NaAc-HAc buffer at pH 4.0 as a solvent: 0.1mM, 0.2mM, 0.3mM, 0.4mM, 0.5mM, 0.6mM, 0.7mM, 0.8mM, 0.9mM, 1.0 mM. The platinum (IV) -based monatomic nanoenzyme (Pt-PDAP), TMB and hydrogen peroxide are respectively and uniformly mixed and placed in a dark place, an ultraviolet spectrophotometer is used for measuring the change condition of absorbance within 10s at 652nm, and the experimental results are shown in fig. 5 and fig. 6, so that the platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) can accord with the kinetic mechanism of peroxidase and can be used as peroxidase. The enzyme kinetic mechanism of the platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) obtained in example 2 conforms to the enzyme kinetic mechanism of peroxidase, and the platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) can be used as peroxidase.
6. Evaluation of Effect as a peroxidase
The specific detection process is as follows:
the platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) prepared in example 2, hydrogen peroxide and a color-developing agent 3,3',5,5' -Tetramethylbenzidine (TMB) were left to stand at room temperature for 30 minutes in the dark, then, the ultraviolet absorbance was measured by an ultraviolet spectrophotometer, and the dependency of platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) on concentration, pH, temperature and hydrogen peroxide concentration was measured, and as a result, as shown in FIGS. 7, 8, 9 and 10, it was revealed that the optimum pH, temperature and hydrogen peroxide concentration were 4.0 to 5.0, 25 to 35 ℃ and 18 to 25mmol/L, respectively, when platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) TMB was developed, and it was demonstrated that the platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) prepared in example 2 could be used as a peroxidase to catalyze low-concentration hydrogen peroxide to generate active oxygen species.
7. And (3) oxygen detection:
the specific detection process is as follows:
the addition of 1MH into platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) with different concentrations is measured by a portable dissolved oxygen measuring instrument 2 O 2 Change of oxygen in the post-solution. When only H is present 2 O 2 Then, as shown in fig. 12, the dissolved oxygen content did not change significantly, and as shown in fig. 11, the oxygen generation rate gradually increased with the increase of the concentration of the platinum (iv) -based monatin nanoenzyme (Pt-PDAP), indicating that the nano-drug has excellent CAT-like activity, i.e., can catalyze H 2 O 2 Production of O 2 。
8. Evaluation of the effect on glutathione:
the specific detection process is as follows: PBS buffer solution with pH 7.4 and concentration of 0.01M is used as solvent, GSH concentration is 50 μ M, and platinum (IV) group monoatomic nanoenzyme (Pt-PDAP) with different concentrations of 10 μ g/mL-100 μ g/mL is shaken for reaction for 3 hours at 30 ℃. The glutathione loss is measured by using an ultraviolet spectrophotometer and a glutathione kit, and compared with a blank control group figure 14 of PBS buffer solution, the result is shown in figure 13, the platinum (IV) group monoatomic nanoenzyme (Pt-PDAP) can effectively reduce the glutathione content, which shows that the platinum (IV) group monoatomic nanoenzyme (Pt-PDAP) can react with glutathione to reduce the GSH content and reduce the loss of active oxygen substances.
9. Animal hyperthermia/radiotherapy co-therapy:
dividing experimental animals into eight groups according to different treatment modes, wherein each group comprises four animals: no treatment control group, II: NIR, III: X-ray, IV: X-ray +808nm, V: material, VI: material + NIR, VII: material + X-ray, VIII: material + NIR + X-ray, NIR: near infrared light. Material usage and concentration: 20 μ L, 2 mg/mL. 808nm near infrared power: 1W/cm 2 And 10 min. Radiotherapy X-ray: 8 Gy. After treatment, the tumor growth tendency of the mice was observed and photographs were taken. Tumor volume and body weight were recorded every other day for each mouse in each group and tumor section observations were made. During the experimental treatment, the tumor temperature was monitored with a thermal infrared imager.
As shown in FIG. 15, which is a bar graph of the cell survival rate of 4T1 cells incubated with a platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) prepared in the present invention, under different treatment protocols, it can be seen that the platinum (IV) -based monatomic nanoenzyme +808nm near infrared light + X-ray has the best killing effect on 4T1 cells, while the untreated control group hardly kills 4T1 cells.
FIG. 16 is a digital photograph showing the tumor change of a tumor-bearing mouse treated by different treatment methods after the platinum (IV) -based monatomic nanoenzyme (Pt-PDAP) prepared in example 2 of the present invention is locally injected into the tumor tissue.
As can be seen in FIG. 16, compared with the case of using only 808nm near-infrared laser irradiation, the platinum (IV) group monatomic nanoenzyme + laser group clearly observed the tumor burn trace, which reflected the effect of thermal therapy. Similarly, the introduction of platinum (IV) -based monatin nanoenzymes (Pt-PDAP) can better inhibit T4 tumor cells than can be achieved using radiotherapy alone. However, compared to the above groups, the tumor suppression effect of the platinum (IV) -based monatomic nanoenzyme +808nm near-infrared light + X-ray group mice was the best, which represents an excellent hyperthermia/radiotherapy synergistic therapeutic performance of the platinum (IV) -based monatomic nanoenzyme (Pt-PDAP).
In conclusion, the invention discloses a preparation method of platinum (IV) group monatomic nanoenzyme, which takes cisplatin and 3, 4-diaminopyridine as precursors to prepare a platinum (IV) group monatomic nanoenzyme medicine through oxidative polymerization. Compared with the prior art, the platinum (IV) group monatomic nanoenzyme medicine prepared by the invention has catalase-like and peroxidase-like activities, can greatly relieve tumor hypoxia, consumes glutathione overexpressed in tumors, can promote the generation of free radicals in tumor cells, reduces toxic and side effects, and can generate hydroxyl free radicals for chemokinetic treatment and improve the treatment effect of radiotherapy. The nano enzyme has strong absorption in the whole near infrared region, can absorb 808nm laser and convert the laser into high-energy heat when being used as a photo-thermal conversion agent, can enhance the dosage of local radiotherapy, greatly improves the killing power on tumor cells and realizes photo-thermal treatment, so that the thermal treatment and the radiotherapy functions of the platinum (IV) -based monoatomic nano enzyme medicament have strong killing effect on the tumor cells in a synergistic manner, and the platinum (IV) -based monoatomic nano enzyme medicament is expected to be widely applied to treatment of tumors or other diseases.
Although the present invention has been described with reference to the above embodiments, it should be understood that the present invention is not limited to the above embodiments, and those skilled in the art can make various changes and modifications without departing from the scope of the present invention.
Claims (10)
1. The platinum-based monatomic nanoenzyme is characterized by consisting of cisplatin cis- [ PtCl [ ] 2 (NH 3 ) 2 ]The hydrogen peroxide, the 3, 4-diaminopyridine and the ultrapure water are prepared by oxidative polymerization, and the 3, 4-diaminopyridine is a monomolecular crystal.
2. The platinum-based monatomic nanoenzyme of claim 1, wherein the nanoenzyme has a particle size of 5nm to 200nm, and the nanoenzyme particles have a fusiform irregular polyhedral structure.
3. A preparation method of platinum-based monatomic nanoenzyme is characterized by comprising the following steps:
s1, adding cisplatin and hydrogen peroxide into ultrapure water, stirring to obtain a solution I, and sequentially recrystallizing, filtering, washing and drying the solution I to obtain powder A;
s2, dissolving the powder A in hydrochloric acid, and stirring to obtain a solution II;
s3, adding ultrapure water into the solution II, then adding 3, 4-diaminopyridine, ultrasonically dissolving, and stirring for reacting to obtain a solution III;
s4, dialyzing and purifying the solution III, and then freeze-drying to obtain a solid sample material;
and S5, performing index detection on the solid sample obtained in the step S4.
4. The method according to claim 3, wherein the molar ratio of cisplatin, hydrogen peroxide, and ultrapure water in step S1 is 1: 102: 417; the stirring speed is 600-800 r/min, the stirring temperature is 50-60 ℃, and the stirring time is 1-1.2 h.
5. The method according to claim 3, wherein the molar ratio of cisplatin, hydrogen peroxide, and ultrapure water in step S1 is 1: 105:420.
6. The method for preparing platinum-based monatomic nanoenzyme according to claim 3, wherein the stirring manner in step S2 is: firstly stirring at the rotating speed of 400-600 r/min for 1-1.5h at the temperature of 40-60 ℃, and then stirring at the rotating speed of 400-600 r/min for 6-18h at normal temperature.
7. The method of claim 3, wherein the hydrochloric acid solution has a concentration of 10mol/L in the step S2.
8. The method for preparing the platinum-based monatomic nanoenzyme of claim 3, wherein the ultrasonic frequency in the step S3 is 55-75 kHz, the reaction condition is that the rotation speed is 600-800 r/min, the reaction is continuously carried out for 24-26h at 37-40 ℃, and the molar ratio of the solution II to the 3, 4-diaminopyridine is 1: 3.
9. The method of claim 3, wherein the dialysis in step S4 is performed by using a dialysis bag with a molecular weight of 12.0kDa to perform dialysis purification, wherein the dialysate is deionized water, the dialysis temperature is 25-35 ℃, and the deionized water is replaced every 2-3 hours and repeated for 3 times.
10. Use of a platinum-based monatomic nanoenzyme according to any of claims 1 to 2 or of a platinum-based monatomic nanoenzyme prepared by the preparation method according to any of claims 3 to 9, for the preparation of a synergic drug for the functional assistance of tumor hyperthermia and radiotherapy.
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