CN114306628A - PEG-MAL modification-based two-dimensional black phosphorus hybrid nano-drug targeting HER2 positive breast cancer - Google Patents

PEG-MAL modification-based two-dimensional black phosphorus hybrid nano-drug targeting HER2 positive breast cancer Download PDF

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CN114306628A
CN114306628A CN202111526452.XA CN202111526452A CN114306628A CN 114306628 A CN114306628 A CN 114306628A CN 202111526452 A CN202111526452 A CN 202111526452A CN 114306628 A CN114306628 A CN 114306628A
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周本青
刘锦杏
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Shantou University
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Abstract

The invention discloses a PEG-MAL modified two-dimensional black phosphorus hybrid nano-drug targeting HER2 positive breast cancer, belonging to the field of nano-drugs. The preparation method comprises the following steps: (1) preparing two-dimensional black phosphorus quantum dots BPQDs with the particle size of 3-7 nm; (2) BP-PEG-MAL-Tmb. The invention selects the novel two-dimensional black phosphorus nano material capable of completely metabolizing as the inorganic nano platform, and has higher drug loading rate and photo-thermal conversion rate. Modification of PEG-MAL can not only improve colloidal stability and biocompatibility of nano-drug, but also effectively enhance capture ability of antigen presenting cells to antigen, and stimulate organism immunoreaction. The modification of Tmb enables the nano-drug to effectively target HER2 positive breast cancer, and further improves the colloidal stability of the nano-drug. The medicine Tmb for resisting HER2 positive breast cancer is organically combined with the two-dimensional black phosphorus for the first time, the targeted photo-thermal immune composite treatment for HER2 positive breast cancer is realized, the design method is simple, the reaction condition is mild, and a new strategy is provided for the accurate treatment of the breast cancer.

Description

PEG-MAL modification-based two-dimensional black phosphorus hybrid nano-drug targeting HER2 positive breast cancer
Technical Field
The invention belongs to the field of nano-drugs, and particularly relates to a PEG-MAL modified two-dimensional black phosphorus hybrid nano-drug targeting HER2 positive breast cancer and a preparation method thereof.
Background
Nanomedicine has become an important development direction in the modern medical field, and the FDA has approved over 60 drug applications containing nanomaterials (e.g., nanoparticles, nanospheres, nanocapsules, and quantum dots), and emerging two-dimensional materials for therapeutic applications have been extensively studied, including Graphene (GO), Black Phosphorus (BP), 15 Transition Metal Dihalides (TMD), 16 transition metal carbides and nitrides. The two-dimensional black phosphorus is used as a novel metal-free two-dimensional nano material, has high biocompatibility and drug loading rate, has light absorption in ultraviolet and near infrared regions, has very high photo-thermal conversion efficiency, and is applicable to the field of biomedicine. However, two-dimensional black phosphorus is very susceptible to degradation when exposed to air or water, Wan et al prepared polyethylene glycol (PEG) modified BP NSs, significantly improving the biocompatibility and physiological stability of BP NSs (J Mater Chem B,8,32,2020, 7076-7120).
Cancer immunotherapy has become a powerful new strategy in cancer treatment. Currently, cancer immunotherapy mainly includes checkpoint inhibitor methods (e.g., anti-PD-1/PD-L1 and anti-CTLA-4 methods), T cell adoptive immunotherapy, vaccine methods, and the like, which have shown good therapeutic effects clinically. Immunotherapy has a slow immune response rate due to the lack of specific targeted therapy analysis, and immunotherapy alone has difficulty in completely ablating localized tumors (Nat Rev Cancer,12,4,2012, 265-. Maleimide (MAL) -modified AC-NPs (Antigen-capturing nanoparticles) can bind to proteins by forming stable thioether bonds, and thus can capture tumor-derived protein antigens (TDPA) and transport them to Antigen Presenting Cells (APC), thereby significantly improving the therapeutic effect of immunotherapy (Nat Nanotechnol,12,9,2017, 877-.
Breast cancer is a highly heterogeneous disease, and different molecular subtype breast cancers have different biological behaviors and clinical pathological characteristics, and the treatment strategies and prognosis of the breast cancers are also greatly different. Currently, breast cancer is generally classified into four molecular subtypes (Nat Rev Clin Oncol,13,11,2016, 674-one 690) according to the conditions of Estrogen Receptor (ER), Progesterone Receptor (PR), human epidermal growth factor receptor 2 (HER 2), and the like. Overexpression of HER2 marked rapid tumor cell proliferation, and Trastuzumab (Trastuzumab, abbreviated as Tmab), a humanized monoclonal antibody that binds to HER2, improved HER2 breast cancer patient prognosis. Tmb in combination with the chemotherapeutic drug paclitaxel or after chemotherapy significantly improves survival and reduces the risk of tumor recurrence (N Engl J Med,372,2, 2015).
The results of the documents and patents in the aspect of searching the hybrid nano-drugs at home and abroad show that: at present, no report on the application aspects of preparation and the like of a PEG-MAL modified two-dimensional black phosphorus hybrid nano-medicament targeting HER2 positive breast cancer is found.
Disclosure of Invention
The invention aims to solve the problems of incomplete treatment effect, easy relapse, immunosuppression and the like of the existing photothermal treatment and immunotherapy.
In order to solve the technical problems, the invention adopts the following technical scheme:
a two-dimensional black phosphorus hybrid nano-drug based on PEG-MAL modification comprises BPQDs modified by PEG-MAL and Tmab.
PEG-MAL (polyethylene glycol-maleimide) and Tmab (trastuzumab) are used for modifying BPQDs (black phosphorus quantum dots), and the PEGylated two-dimensional black phosphorus nano hybrid drug modified with trastuzumab is used for targeting HER2 positive metastatic breast cancer, so that the photothermal immunotherapy effect is achieved. It was found that NH2The PEG-MAL is heterobifunctional polyethylene glycol containing maleimide and amino, can be used for modifying proteins, polypeptides and other materials or small molecules containing sulfydryl, can enhance the capture capacity of antigen presenting cells to antigens by the MAL, can effectively target HER2 positive breast cancer by trastuzumab, and can be used as an effective photothermal reagent by two-dimensional black phosphorus. The invention uses two-dimensional black phosphorus as a carrier to successfully modify PEG-MAL andtab, the photothermal therapy and the immunotherapy are organically combined, so that a better treatment effect is achieved, and the aim of eliminating tumors more thoroughly is achieved by exciting the immune regulation of an organism while local tumor ablation is realized.
A preparation method of the PEG-MAL modification-based two-dimensional black phosphorus hybrid nano-drug comprises the following steps:
A. dissolving the blocky black phosphorus crystal powder in an amide organic solvent, centrifuging and washing after ultrasonic oscillation and crushing to obtain a BPQDs aqueous solution;
B. mixing the aqueous solution of BPQDs with NH2And uniformly mixing the PEG-MAL aqueous solution and the Tmb aqueous solution to obtain a mixed solution, then carrying out ultrasonic treatment, stirring and centrifuging to obtain the PEG-MAL modification-based two-dimensional black phosphorus hybrid nano-drug.
Preferably, in the mixed solution, BPQDs, NH2-PEG-MAL, Tmab molar ratio 1: 5-10: 2 to 6.
Preferably, the concentration of the BPQDs aqueous solution is 5-10 mg/mL; the NH2NH in aqueous PEG-MAL solution2The molecular weight of the-PEG-MAL is 2000.
Preferably, the BPQDs aqueous solution is 3-7nm two-dimensional black phosphorus quantum dots.
The BPQDs aqueous solution is 3-7nm two-dimensional black phosphorus quantum dots, the concentration is 5-10 mg/mL, and the volume dosage is about 500-1000 muL. NH (NH)2The molecular weight of the-PEG-MAL is 2000, and the mass dosage is about 15-60 mg. The mass dosage of Tma is about 7.5-30 mg.
Preferably, the mass-to-volume ratio of the massive black phosphorus crystal powder to the NMP is 50-100 mg: 20-30 mL.
Preferably, the method comprises the following steps:
A. dissolving the blocky black phosphorus crystal powder in the NMP (1-methyl-2-pyrrolidone), firstly performing ultrasonic oscillation crushing by a probe, then performing ultrasonic oscillation cleaning in a low-temperature water bath, then centrifuging at 3500-4500 rpm for 10-15 min to remove large blocky precipitates, then centrifuging at 13000-14000 rpm and washing with deionized water to obtain the uniformly dispersed BPQDs aqueous solution;
B. mixing the aqueous solution of BPQDs with NH2-PEG-MAL aqueous solution, Tmab aqueous solution is uniformly mixed to obtain the mixed solution, then water bath ultrasound is carried out for 20-30 min, then the mixed solution is stirred at the room temperature of 400-500 rpm in a dark place for 10-12 h, and the mixture is centrifuged at the rotating speed of 13000-14000 rpm for 10-15 min to obtain the PEG-MAL modification-based two-dimensional black phosphorus hybrid nano-drug.
In step A, the black phosphorus crystal powder can be dissolved in amide organic solvents, and amide organic solvents similar to NMP can be used as dispersants for BP, but the BP has the best dispersion stability in NMP and is considered to be a better choice.
Preferably, in the step a, the specific operations of firstly performing ultrasonic oscillation crushing on the probe and then performing ultrasonic oscillation cleaning in the low-temperature water bath are as follows: firstly, under the circulating condition that the power is 810W and the ultrasonic frequency is 2.0s for 1.0s, an ultrasonic cell crusher with a No. 6 horn is used for carrying out ice bath ultrasonic crushing for 40-50 min, repeating for 3-5 times, and then carrying out water bath ultrasonic cleaning for 1-2 h at the temperature of 0-10 ℃.
Because the black phosphorus is extremely unstable and is extremely easy to decompose under the condition of high temperature, the black phosphorus is carried out under the condition of ice bath in the ultrasonic crushing process, and the cleaning process also ensures low temperature, and the temperature is about 0-10 ℃ approximately. The two-dimensional black phosphorus quantum dots with uniform dispersion of 3-7nm can be obtained by firstly carrying out ultrasonic oscillation crushing on the probe and then carrying out ultrasonic oscillation cleaning in a low-temperature water bath.
An application of the PEG-MAL modification-based two-dimensional black phosphorus hybrid nano-drug in preparation of a cancer photo-thermal immune composite treatment targeted drug.
Preferably, the cancer is HER2 positive breast cancer.
The invention can be directly used for the photothermal immune complex treatment of cancer or the preparation of related medicaments, in particular to the photothermal immune complex treatment of HER2 positive metastatic breast cancer.
The implementation of the invention has the following beneficial effects:
(1) the preparation method is simple and easy to operate, the uniformly dispersed two-dimensional black phosphorus quantum dots are obtained by utilizing a liquid phase stripping method combining probe ultrasound and water bath ultrasound, the PEG-MAL and Tma are successfully modified by utilizing the electrostatic adsorption effect of mutual attraction of positive and negative charges, and the prepared hybrid nano-drug has good targeting property on HER2 positive breast cancer, and simultaneously, the photo-thermal treatment and the immunotherapy are organically combined, so that the preparation method has good application prospect;
(2) the PEG-MAL is modified on the surface of the two-dimensional black phosphorus, the capture capacity of antigen presenting cells to antigen can be effectively enhanced, the immunocompetence of an organism is greatly enhanced, Tma is used as a known medicine for HER2 over-expressed metastatic breast cancer, HER2 positive breast cancer can be effectively targeted, the prepared hybrid nano medicine can enhance the immune function of the organism to achieve the effect of more thoroughly eliminating tumors while performing photothermal therapy, and the invention lays a good foundation for a novel photothermal immune combination therapy of HER2 positive breast cancer.
(3) The invention selects the novel two-dimensional black phosphorus nano material capable of completely metabolizing as the inorganic nano platform, and has higher drug loading rate and photo-thermal conversion rate. Modification of PEG-MAL can not only improve colloidal stability and biocompatibility of nano-drug, but also effectively enhance capture ability of antigen presenting cells to antigen, and stimulate organism immunoreaction. The modification of Tmb enables the nano-drug to effectively target HER2 positive breast cancer, and further improves the colloidal stability of the nano-drug. The medicine Tmb for resisting HER2 positive breast cancer is organically combined with the two-dimensional black phosphorus for the first time, the targeted photo-thermal immune composite treatment for HER2 positive breast cancer is realized, the design method is simple, the reaction condition is mild, and a new strategy is provided for the accurate treatment of the breast cancer.
Drawings
FIG. 1 is an analytical view of prepared BPQDs, in which: (a) TEM images of BPQDs; (b) HRTEM image of BPQDs; (c) AFM images of BPQDs; (d) EDS diagrams of BPQDs.
FIG. 2 is an analytical chart of the prepared BP-PEG-MAL-Tmb, wherein: (a) is a surface potential histogram of BP-PEG-MAL-Tmb; (b) is a histogram of the particle size of BP-PEG-MAL-Tmb.
FIG. 3 shows the laser irradiation (1.0W/cm) at 808nm of BP-PEG-MAL-Tmb with different concentrations2) Temperature rise profile of the following.
FIG. 4 shows that 50. mu.g/ml BP-PEG-MAL-Tma is at 1W/cm2808nm laserTemperature rise-temperature drop curve under irradiation.
FIG. 5 is a graph showing the ultraviolet absorption spectra of BPQDs at day 0, day 7 and day 15 in the range of 450 to 900 nm.
FIG. 6 is a graph of the ultraviolet absorption spectrum of BP-Tmb in the range of 450-900 nm on days 0,7 and 15.
FIG. 7 is a graph showing the relative cell viability after incubation of different concentrations of BP-PEG-MAL-Tmab with MDA-MB-231 cells for 24h, 48h and 72h, i.e., the dark toxicity evaluation of MDA-MB-231 cells according to the present invention.
FIG. 8 is a graph showing the relative cell survival rates of BP-PEG-MAL-Tmab with different concentrations after incubation for 24h, 48h and 72h with SKBR3 cells, namely the dark toxicity evaluation of SKBR3 cells according to the invention.
FIG. 9 shows that BP-PEG-MAL-Tmab with different concentrations was co-cultured with MDA-MB-231 cells and SKBR3 cells respectively for 6h, and then irradiated with 808nm laser for 10min (1W/cm)2) And then, after incubation for 2 hours, the relative cell survival rate is shown as a schematic diagram of the targeted phototoxicity evaluation of different cells.
FIG. 10 shows that different concentrations of BP-PEG-MAL-Tmab and MDA-MB-231 cells were co-cultured for 6h and then irradiated with 808nm laser for 10min (1W/cm)2) And then, the staining pattern of the live and dead cells after further incubation for 2h is a schematic diagram of the phototoxicity evaluation of MDA-MB-231 cells by the invention.
FIG. 11 shows that BP-PEG-MAL-Tmab with different concentrations and SKBR3 cells were co-cultured for 6h and then irradiated with 808nm laser for 10min (1W/cm)2) And then, after further incubation for 2h, a staining pattern of the live and dead cells is shown, namely a schematic diagram of the phototoxicity evaluation of SKBR3 cells by the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail with reference to the accompanying drawings.
Example 1
The preparation of BPQDs comprises the following steps:
(1) the preparation of the two-dimensional black phosphorus quantum dot adopts a liquid phase stripping method. Dissolving blocky black phosphorus crystal powder in 1-methyl-2-pyrrolidone, performing ice-bath ultrasonic crushing for 45min by using an ultrasonic cell crusher with a No. 6 horn, repeating for 3 times, wherein the power of the ultrasonic cell crusher is 810W, the ultrasonic frequency is 2.0s on and 1.0s off, and performing ultrasonic cleaning for 1h in a 4 ℃ low-temperature water bath;
(2) and then placing the mixture into a centrifuge, centrifuging at 4000rpm for 12min to remove large blocky particle precipitates, then centrifuging at 13000rpm for 12min to obtain the two-dimensional black phosphorus quantum dots required by people, then adding deionized water, centrifuging at 13000rpm again for 12min to remove residual NMP to obtain a BPQDs aqueous solution, quantifying the black phosphorus concentration by using an ultraviolet-visible spectrophotometer, and storing at 4 ℃ for later use.
The preparation of BP-PEG-MAL-Tmb NPs comprises the following steps:
adding aqueous solution of BPQDs, NH2Mixing the aqueous solution of the PEG-MAL and the aqueous solution of the Tmb, performing water bath ultrasound for 20min, then stirring at 450rpm in the dark for 10h at room temperature, and centrifuging (13000rpm for 10min) to remove the excess NH not bound2Washing the PEG-MAL and the Tmb by using deionized water, and collecting all precipitates to obtain the BP-PEG-MAL-Tmb hybridized nano-drug.
Example 2
The preparation of BPQDs comprises the following steps:
(1) the preparation of the two-dimensional black phosphorus quantum dot adopts a liquid phase stripping method. Dissolving blocky black phosphorus crystal powder in 1-methyl-2-pyrrolidone, performing ice-bath ultrasonic crushing for 40min by using an ultrasonic cell crusher with a No. 6 horn for 5 times, wherein the power of the ultrasonic cell crusher is 810W, the ultrasonic frequency is 2.0s on and 1.0s off, and then performing ultrasonic cleaning for 2h in a low-temperature water bath at 0 ℃;
(2) and then placing the mixture into a centrifuge, centrifuging at 4500rpm for 10min to remove large blocky particle precipitates, then centrifuging at 14000rpm for 10min to obtain the two-dimensional black phosphorus quantum dots required by people, then adding deionized water, centrifuging at 14000rpm for 10min again to remove residual NMP to obtain BPQDs aqueous solution, quantifying the black phosphorus concentration by using an ultraviolet-visible spectrophotometer, and storing at 0 ℃ for later use.
The preparation of BP-PEG-MAL-Tmb NPs comprises the following steps:
adding aqueous solution of BPQDs, NH2Mixing the aqueous solution of-PEG-MAL and the aqueous solution of Tmab, performing ultrasonic treatment in water bath for 30min, and then performing ultrasonic treatment in roomStirring at 400rpm at room temperature for 12h in the dark, and centrifuging (14000rpm, 15min) to remove excess NH unbound2Washing the PEG-MAL and the Tmb by using deionized water, and collecting all precipitates to obtain the BP-PEG-MAL-Tmb hybridized nano-drug.
Example 3
The preparation of BPQDs comprises the following steps:
(1) the preparation of the two-dimensional black phosphorus quantum dot adopts a liquid phase stripping method. Dissolving blocky black phosphorus crystal powder in 1-methyl-2-pyrrolidone, performing ice-bath ultrasonic crushing for 50min by using an ultrasonic cell crusher with a No. 6 horn for 3 times, wherein the power of the ultrasonic cell crusher is 810W, the ultrasonic frequency is 2.0s on and 1.0s off, and then performing ultrasonic cleaning for 1h in a 10 ℃ low-temperature water bath;
(2) and then placing the mixture into a centrifuge, centrifuging at 3500rpm for 15min to remove large blocky particle precipitates, centrifuging at 13000rpm for 15min to obtain the two-dimensional black phosphorus quantum dots required by people, adding deionized water, centrifuging at 13000rpm again for 15min to remove residual NMP to obtain a BPQDs aqueous solution, quantifying the black phosphorus concentration by using an ultraviolet-visible spectrophotometer, and storing at 2 ℃ for later use.
The preparation of BP-PEG-MAL-Tmb NPs comprises the following steps:
adding aqueous solution of BPQDs, NH2Mixing the aqueous solution of the PEG-MAL and the aqueous solution of the Tmb, performing water bath ultrasound for 20min, then stirring at 500rpm at room temperature in the dark for 10h, and centrifuging (13000rpm for 10min) to remove the excess NH not bound2Washing the PEG-MAL and the Tmb by using deionized water, and collecting all precipitates to obtain the BP-PEG-MAL-Tmb hybridized nano-drug.
Effect example 1
Taking example 1 as an example, the prepared BPQDs are observed, and 2D BPQDs are found to be uniformly dispersed and have the size of about 3-7nm through observation (see figure 1a), HRTEM images show clear lattice stripes (see figure 1b), AFM images show that the height of the BPQDs is about 5nm (see figure 1c), and EDS images further confirm the existence of phosphorus elements (see figure 1D). Dynamic light scattering technique for BPQDs, NH2Surface potential (see FIG. 2a) and hydration of-PEG-MAL, Tma, BP-PEG-MAL, BP-Tma, BP-PEG-MAL-TmaParticle size analysis (see FIG. 2b), by comparing the potential of BPQDs and particle size, we can see that the final material BP-PEG-MAL-Tmb prepared by us successfully loads NH on the material2PEG-MAL and Tmb. BP-PEG-MAL-Tmb with different concentrations is irradiated by laser at 808nm for 10min (1W/cm)2) Near infrared light can be rapidly converted into heat energy (see fig. 3). For example, at a concentration of 100ppm, the temperature of BP-PEG-MAL-Tma reached around 50 ℃, whereas the temperature of PBS of the control group reached only 30 ℃ under the same conditions. Tumor cells are known to be very sensitive to temperature, and have killing ability when the temperature is higher than 42 ℃. In addition, after 5 laser on/off cycles, no obvious attenuation is seen in the temperature rising capability, which indicates that the hybrid nano-drug shows good photo-thermal stability (see fig. 4).
Effect example 2
According to the conditions of the example 1, aqueous solutions of BPQDs and BP-Tma are respectively prepared and placed at room temperature, then ultraviolet absorption spectra of 0 th day, 7 th day and 15 th day of the BPQDs and the BP-Tma in the range of 450-900 nm are respectively measured by an ultraviolet-visible spectrophotometer (see a figure 5 and a figure 6), and compared with a BPQDs group, the BP-Tma improves the stability of the BPQDs obviously. Table 1 shows the absorbance values at 700nm for BPQDs and BP-Tma at days 0,7 and 15 and the decay values at day 15 compared to day 0. Taking day 15 as an example (see table 1), the absorbance of the BPQDs group at 700nm is reduced by 41.82% compared with day 0, while the absorbance of the BP-Tma group is reduced by only 6.34%, which effectively proves that the stability of BP can be obviously improved and the loss caused by oxidative decomposition can be reduced after the Tma is modified.
Table 1
Figure BDA0003410642280000091
Effect example 3
Taking the BP-PEG-MAL-Tma hybrid nano-drug obtained in example 1 as an example, cytotoxicity of BP-PEG-MAL-Tma NPs with different concentrations on MDA-MB-231 cells (shown in figure 7) and SKBR3 cells (shown in figure 8) was evaluated by a CCK-8 method, and taking co-culture for 24h as an example, the nano-hybrid drug has one effect on two breast cancer cellsThe toxicity of SKBR3 is slightly higher than that of MDA-MB-231 cells, which is related to the targeting of the nano hybrid drug to HER2 positive breast cancer cells, and the cytotoxicity is higher as the co-culture time is longer. Double staining of live and dead cells further confirmed the above conclusion (see FIGS. 9, 10, 11), and Calcein-AM was known to stain live cells and PI-stained cells, and laser irradiation was given at 100ppm + L (+ L, 1W/cm)2) For example, after laser irradiation for 10min, the ratio of MDA-MB-231 cell living cells is 11%, while SKBR3 cell living cells are only 4%, and the targeting property and the good photothermal conversion capability of the nano hybrid drug are proved again.
While the invention has been described in connection with what is presently considered to be the most practical and preferred embodiment, it is to be understood that the invention is not to be limited to the disclosed embodiment, but on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims.

Claims (10)

1. A two-dimensional black phosphorus hybrid nano-drug based on PEG-MAL modification is characterized by comprising BPQDs modified by PEG-MAL and Tmb.
2. The preparation method of the PEG-MAL modification-based two-dimensional black phosphorus hybrid nano-drug according to claim 1, which comprises the following steps:
A. dissolving the blocky black phosphorus crystal powder in an amide organic solvent, centrifuging and washing after ultrasonic oscillation and crushing to obtain a BPQDs aqueous solution;
B. mixing the aqueous solution of BPQDs with NH2And uniformly mixing the PEG-MAL aqueous solution and the Tmb aqueous solution to obtain a mixed solution, then carrying out ultrasonic treatment, stirring and centrifuging to obtain the PEG-MAL modification-based two-dimensional black phosphorus hybrid nano-drug.
3. The preparation method of the PEG-MAL modification-based two-dimensional black phosphorus hybrid nano-drug according to claim 2, wherein in the mixed solution, BPQDs, NH2-PEG-MAL, Tmab molar ratio 1: 5-10: 2 to 6.
4. According to claim 2The preparation method of the PEG-MAL modified two-dimensional black phosphorus hybrid nano-drug is characterized in that the concentration of the BPQDs aqueous solution is 5-10 mg/mL; the NH2NH in aqueous PEG-MAL solution2The molecular weight of the-PEG-MAL is 2000.
5. The preparation method of the PEG-MAL modified two-dimensional black phosphorus hybrid nano-drug according to claim 2, wherein the BPQDs aqueous solution contains 3-7nm of two-dimensional black phosphorus quantum dots.
6. The preparation method of the PEG-MAL modified two-dimensional black phosphorus hybrid nano-drug as claimed in claim 2, wherein the mass-to-volume ratio of the bulk black phosphorus crystal powder to the NMP is 50-100 mg: 20-30 mL.
7. The preparation method of the PEG-MAL modification-based two-dimensional black phosphorus hybrid nano-drug according to claim 2, which comprises the following steps:
A. dissolving the blocky black phosphorus crystal powder in the NMP, firstly carrying out ultrasonic oscillation crushing by a probe, then carrying out ultrasonic oscillation cleaning in a low-temperature water bath, then centrifuging at 3500-4500 rpm for 10-15 min to remove large blocky precipitates, then centrifuging at 13000-14000 rpm, and washing with deionized water to obtain the uniformly dispersed BPQDs aqueous solution;
B. mixing the aqueous solution of BPQDs with NH2And (3) uniformly mixing a PEG-MAL aqueous solution and a Tmb aqueous solution to obtain the mixed solution, performing water bath ultrasound for 20-30 min, stirring at 400-500 rpm in a dark place for 10-12 h at room temperature, and centrifuging at 13000-14000 rpm for 10-15 min to obtain the PEG-MAL modification-based two-dimensional black phosphorus hybrid nano-drug.
8. The preparation method of the PEG-MAL modification-based two-dimensional black phosphorus hybrid nano-drug as claimed in claim 7, wherein in the step A, the specific operations of probe ultrasonic oscillation crushing and low-temperature water bath ultrasonic oscillation cleaning are as follows: firstly, under the circulating condition that the power is 810W and the ultrasonic frequency is 2.0s for 1.0s, an ultrasonic cell crusher with a No. 6 horn is used for carrying out ice bath ultrasonic crushing for 40-50 min, repeating for 3-5 times, and then carrying out water bath ultrasonic cleaning for 1-2 h at the temperature of 0-10 ℃.
9. The application of the PEG-MAL modification-based two-dimensional black phosphorus hybrid nano-drug according to claim 1 in the preparation of a cancer photothermal immune complex therapy targeted drug.
10. The use of the PEG-MAL modification-based two-dimensional black phosphorus hybrid nanopharmaceutical as claimed in claim 9, wherein the cancer is HER2 positive breast cancer.
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