CN111888333A - Transferrin receptor targeted nano micelle and preparation method and application thereof - Google Patents
Transferrin receptor targeted nano micelle and preparation method and application thereof Download PDFInfo
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- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
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- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
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Abstract
The invention discloses a transferrin receptor targeted nano micelle and a preparation method and application thereof, wherein the nano micelle comprises TfT-T12 short peptide and polyethylene glycol-polylactic acid connected with the TfT-T12 short peptide through amido bond. The invention utilizes the characteristics of good biological safety, biocompatibility and the like of TfT-T12-PEG-PLA and contains a hydrophilic part and a hydrophobic part, so that the nano-micelle can be formed by self-assembly in water, has stronger transferrin targeting property, and can be used as a drug carrier to carry drugs to cross a blood brain barrier, therefore, the nano-micelle can be used as a transferrin targeting carrier and can be widely applied to the field of treatment of brain glioma.
Description
Technical Field
The invention relates to the technical field of drug carriers, in particular to a transferrin receptor targeted nano micelle and a preparation method and application thereof.
Background
Brain glioma is a common intracranial primary tumor, and because of high malignancy, strong invasiveness and high recurrence rate, the median survival time of a patient is still less than 14 months and the 5-year mortality rate is as high as 95 percent despite the comprehensive treatment of surgery, radiotherapy, chemotherapy and the like. For the glioma with high malignancy, the first-line treatment scheme adopted clinically is combined radiotherapy and chemotherapy after surgical resection, and the postoperative chemotherapy is an important treatment means for the brain glioma, but the Blood Brain Barrier (BBB) and the drug resistance of hydrophobic chemotherapy drugs seriously affect the effect of chemotherapy, so the treatment of the brain glioma is still a worldwide acknowledged problem. The blood brain barrier is a natural barrier that protects brain tissue from metabolic products, maintaining central nervous system homeostasis. However, while protecting the nervous system, the blood-brain barrier also prevents nearly 100% of large molecule drugs and 98% of small molecule drugs from reaching the brain tissue efficiently, resulting in most central nervous system diseases, such as malignant brain tumors, being ineffectively treated. Therefore, for intracerebral diseases such as malignant brain tumors, the greatest challenge to current clinical treatment is how to effectively pass the drug across the blood-brain barrier.
Accordingly, the prior art is yet to be improved and developed.
Disclosure of Invention
In view of the defects of the prior art, the invention aims to provide a transferrin receptor targeted nano micelle, a preparation method and an application thereof, and aims to solve the problem that the existing drugs cannot effectively penetrate through a blood brain barrier.
The technical scheme of the invention is as follows:
a transferrin receptor targeted nanomicelle, which comprises TfT-T12 short peptide and polyethylene glycol-polylactic acid connected with the TfT-T12 short peptide through amido bond.
The transferrin receptor targeted nano micelle is characterized in that the amino acid structural sequence of the TfT-T12 short peptide is THRPPMWSPVWP.
The transferrin receptor targeted nano micelle is characterized in that the particle size of the nano micelle is 100-200 nm.
The preparation method of the transferrin receptor targeted nano micelle comprises the following steps:
dissolving polyethylene glycol-polylactic acid in an organic solvent, adding 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and benzotriazole for activation treatment to obtain an activated polyethylene glycol-polylactic acid solution;
TfT-T12 short peptide and triethylamine are added into the activated polyethylene glycol-polylactic acid solution, and the nano micelle targeted by the transferrin receptor is generated through reaction.
The preparation method of the transferrin receptor targeted nano micelle comprises the step of preparing a nano micelle by using a solvent, wherein the organic solvent is N, N-dimethylformamide or dimethyl sulfoxide.
The preparation method of the transferrin receptor targeted nano micelle comprises the step of activating for 2-3 h.
The application of the transferrin receptor targeted nano-micelle is characterized in that the transferrin receptor targeted nano-micelle is used for encapsulating hydrophobic chemotherapeutic drugs.
The application of the transferrin receptor targeted nano micelle is characterized in that the drug loading of the transferrin receptor targeted nano micelle is 1-10%.
The transferrin receptor targeted nano micelle is applied, wherein the hydrophobic chemotherapeutic drug is paclitaxel.
The application of the transferrin receptor targeted nano micelle is that the mass ratio of the transferrin receptor targeted nano micelle to paclitaxel is 10: 1.
Has the advantages that: the invention provides a transferrin receptor targeted nano micelle, which comprises TfT-T12 short peptide and polyethylene glycol-polylactic acid connected with the TfT-T12 short peptide through amido bond. The invention utilizes the characteristics of good biological safety, biocompatibility and the like of TfT-T12-PEG-PLA and contains a hydrophilic part and a hydrophobic part, so that the nano-micelle can be formed by self-assembly in water, has stronger transferrin targeting property, and can be used as a drug carrier to carry drugs to cross a blood brain barrier, therefore, the nano-micelle can be used as a transferrin targeting carrier and can be widely applied to the field of treatment of brain glioma.
Drawings
FIG. 1 is a graph showing the results of hydrogen spectroscopy of TfR-T12-PEG-PLA in example 1.
FIG. 2a is a graph showing the results of the particle size measurement of the nanomicelle in example 2.
FIG. 2b is a TEM representation of the nanomicelles of example 2
FIG. 3 is the in vitro release results of the nanomicelle in example 2
Fig. 4a is a graph of the results of the evaluation of the uptake of nanomicelles by cells by flow cytometry in example 3.
FIG. 4b is a graph of the results of the evaluation of the uptake of nanomicelles by cells by confocal laser scanning microscopy in example 3.
FIG. 5 is a graph showing the results of inhibiting tumor cell proliferation by the nanomicelles in example 4.
FIG. 6 is a graph showing the results of the effect of nanomicelles crossing the blood-brain barrier in example 5.
FIG. 7a is a graph of the results of quantifying the tumor growth inhibition effect of the nanomicelle in example 6 on tumor-bearing mice (subcutaneous model).
FIG. 7b is a graph showing the results of the tumor growth inhibition of the nanomicelle in example 6 on tumor-bearing mice (subcutaneous model).
FIG. 8 is a graph showing the results of the inhibition of angiogenesis in tumor-bearing mice by nanomicelle in example 7.
FIG. 9 is a graph showing the effect of nanomicelles in example 8 on the prolongation of the survival cycle of tumor-bearing mice (in situ model).
FIG. 10 is an H & E staining micrograph of heart, liver, spleen, lung and kidney when nano-micelle safety was investigated in example 9.
Detailed Description
The invention provides a transferrin receptor targeted nano micelle and a preparation method and application thereof, and the invention is further described in detail below in order to make the purpose, technical scheme and effect of the invention clearer and more clear and definite. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The transferrin Receptor (TfR) is a glycoprotein present on the cell surface, and is a most representative of transcytosis receptors, which is expressed on various cells of human body, especially on the surface of some tumor cells, such as brain tumor cells and liver tumor cells, and the expression of TfR Receptor is obviously higher than that of normal cells. Besides, receptors such as TfR are simultaneously highly expressed on brain endothelial cells, so that the medicament targeting TfR can pass through a blood brain barrier theoretically, can specifically identify brain glioma cells and can be enriched to malignant brain tumor parts in the brain. Based on the above theory, after being combined with a TfR receptor, a drug modified by a TfR ligand can pass through a blood brain barrier through transcytosis of brain endothelial cells, and in malignant brain tumor cells, the drug modified by the TfR ligand can be specifically taken up by brain glioma cells under the mediation of the receptor TfR, so that the therapeutic drug is specifically enriched to a tumor region and inhibits the growth of malignant brain tumors, thereby achieving the effect of targeted delivery. Therefore, the carrier modified by the TfR ligand is an ideal drug carrier capable of crossing the blood brain barrier and being specifically taken up by brain glioma cells, and is used for targeted delivery of hydrophobic chemotherapeutic drugs.
The polyethylene glycol-polylactic acid (PEG-PLA) nano micelle is composed of amphiphilic block polymers, namely, the polyethylene glycol-polylactic acid nano micelle simultaneously has a lipophilic hydrophobic polylactic acid segment (PLA) and a hydrophilic polyethylene glycol (PEG) segment, after the polyethylene glycol-polylactic acid forms micelle under the self-assembly condition, a hydrophobic inner core can entrap lipophilic medicaments such as paclitaxel, and a hydrophilic outer shell part has better biocompatibility, and meanwhile, the stability of the micelle in an aqueous solution can be improved, and the circulation time of the medicaments in vivo is prolonged. In addition, the nanometer drug-loaded micelle has smaller particle size, which is beneficial to improving the high permeability and retention effect of the solid tumor, namely the so-called EPR effect (enhanced permeability and retention effect), the EPR effect is the specific high permeability and retention effect of the solid tumor tissue, wherein the existence of capillary vascular endothelial cell fissures in the solid tumor causes the high permeability of the macromolecular drug, and in addition, the retention of the macromolecular drug is caused due to the structural integrity of the capillary lymphatic vessel and the damaged characteristics of the lymphatic system in the solid tumor, so that the selective distribution of macromolecules or particles in the tumor tissue is promoted. Therefore, the PEG-PLA nano micelle can solve the problem of solubility of insoluble drugs, and can also enhance the accumulation of the drugs in tumor tissues by utilizing the passive targeting effect on tumors generated by the characteristic EPR effect of the tumor tissues, improve the bioavailability of hydrophobic chemotherapeutic drugs and enhance the killing property of the hydrophobic chemotherapeutic drugs on the tumors.
Based on the above, the invention provides a transferrin receptor targeted nano-micelle, which comprises TfT-T12 short peptide and polyethylene glycol-polylactic acid connected with the TfT-T12 short peptide through amido bond.
The nano micelle provided by the invention can specifically identify TfR, so that the TfR can pass through a blood brain barrier, has a good effect of targeting brain glioma, has good safety, biocompatibility and in-vivo stability, and slowly releases a drug under the condition of simulating an internal environment; the nano micelle and a hydrophobic chemotherapeutic drug form a nano preparation with good stability, and the nano preparation has pharmacological activity and good brain glioma targeting effect.
In some embodiments, the TfT-T12 short peptide consists of 12 amino acids, has a molecular weight of 1490, has an amino acid structure sequence of thrpmwspvwp, and has a receptor highly expressed on brain endothelial cells and brain glioma cells, so that the nano-micelle modified by the TfR-T12 short peptide can smoothly cross the blood brain barrier and specifically recognize the brain glioma cells.
In some embodiments, the nanoparticle size is 100-200 nm.
In some embodiments, there is also provided a method of preparing a transferrin receptor targeted nanomicelle, comprising the steps of:
dissolving polyethylene glycol-polylactic acid in an organic solvent, adding 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and benzotriazole for activation treatment to obtain an activated polyethylene glycol-polylactic acid solution; TfT-T12 short peptide and triethylamine are added into the activated polyethylene glycol-polylactic acid solution, and the nano micelle targeted by the transferrin receptor is generated through reaction.
Specifically, PEG-PLA with a carboxyl end is dissolved in an organic solvent, 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and benzotriazole are added as activating agents, room temperature activation is carried out for 2-3h to obtain activated PEG-PLA, then TfR-T12 short peptide and a small amount of triethylamine are added, reaction is continued for 24h, dialysis is carried out in deionized water for 48h, and freeze drying is carried out for 24h to obtain TfR-T12-PEG-PLA, wherein the synthetic route is as follows:
further, dissolving the TfR-T12-PEG-PLA in an organic solvent, adding a corresponding chemotherapeutic drug according to the mass ratio of the carrier material to the chemotherapeutic drug, slowly dropping the mixed solution into 50ml of deionized water under the condition of slight stirring, dialyzing for 24 hours, removing the drug which is not entrapped by a 0.45 mu m microporous filter membrane, and freeze-drying for 24 hours to obtain the drug-loaded nano micelle.
In some embodiments, the organic solvent is N, N-dimethylformamide or dimethylsulfoxide, but is not limited thereto.
In some embodiments, there is also provided a use of a transferrin receptor targeted nanomicelle for encapsulating a hydrophobic chemotherapeutic drug.
In some embodiments, the transferrin receptor targeted nanomicelle drug load is 1-10%.
In some specific embodiments, the hydrophobic chemotherapeutic is paclitaxel, and preferably, the mass ratio of the transferrin receptor targeted nanomicelle to paclitaxel is 10: 1. In the embodiment, the paclitaxel can form a nano preparation with good stability with the nano micelle, and has pharmacological activity and good brain glioma targeting effect.
The preparation method and the performance of the transferrin nano micelle of the invention are further explained by the following specific examples:
example 1
Synthesis and characterization of TfR-T12-PEG-PLA
50mg HOOC-PEG-PLA copolymer was activated with EDC/NHS (10mg EDC/5.8mg NHS) for 3h, TfR-T12 peptide was added to the DMSO solution, and the reaction was maintained for an additional 24h with moderate stirring. The reacted mixture was then dialyzed in distilled water for 48h (dialysis bag cut-off 3500Da) to remove unbound TfR-T12 peptide, and the dialysate was freeze-dried for 24 h. The structure of TfR-T12-PEG-PLA was characterized by 1H-NMR spectroscopy, and as a result, as shown in FIG. 1, the methylene group of PEG-PLA was detected in the signal of 3.65ppm, and the signals of CH and CH3 from PLA were shown at the 5.3 and 1.6ppm positions, respectively. Chemical shifts of 7.0-8.0ppm are considered to be hydrogen protons in the TfR-T12 peptide chain. The results indicate that the TfR-T12 peptide successfully binds to PEG-PLA.
Example 2
Preparation and characterization of nanomicelle
100mg of TfR-T12-PEG-PLA copolymer and 10mg of PTX were dissolved in 2ml of DMSO, then added dropwise to 50ml of water with moderate stirring, the mixture was stirred at room temperature for 30min and dialyzed in distilled water for 48h (molecular weight cut-off 3500 Da). The non-entrapped PTX was removed by filtration using a 0.45 μ M filter.
The surface morphology of the drug-loaded micelle was observed by a transmission electron microscope. The sample solution of the drug-loaded micelle is prepared to a concentration of 1mg/ml, accurately measured to 20 μ L, and placed on a copper mesh, the copper mesh is placed under a lamp for 10min to be dried, and then the surface morphology of the drug-loaded micelle is observed by a transmission electron microscope.
The release of PTX in vitro was evaluated under physiological conditions for 48h, with free PTX and nanomicelles not modified by TfR-T12 as controls. Briefly, a nanomicelle sample containing 0.45mg of PTX was added to a dialysis bag. Then, the dialysis bag was immersed in 50mL of PBS (pH 7.4 containing 0.1% Tween 80), slowly stirred at 37 ℃, and at a predetermined time point, 0.2mL of a sample was taken out of the medium and the same volume of fresh medium was added, and the concentration of PTX was analyzed with an ultraviolet spectrophotometer.
The average particle diameter of the TfR-T12-PMs is 110nm (polydispersity index: 0.217) (as shown in FIG. 2 a), the electromotive potentials of PEG-PMs and TfR-T12-PMs are-7.91 and-4.77 respectively, the PEG-PMs are negatively charged due to the carboxyl group at the end of polyethylene glycol, and the electromotive potentials of the TfR-T12-PMs are slightly increased along with the modification of the TfR-T12. From the morphology image of the transmission electron microscope (FIG. 2B), the TfR-T12-PMs are spherical and uniformly distributed. The PTX of PEG-PMs and TfR-T12-PMs was released slowly compared to free PTX in PBS at pH 7.4 at 37 ℃, with about 50% of the PTX released from the nanomicelles within 24h and 70% of the free PTX released at the same time point. Therefore, the modification of TfR-T12 has no obvious influence on the drug release of the nano-micelle (as shown in figure 3).
Example 3
Uptake assay of nanomicelle
In order to evaluate the uptake and intracellular localization of nanomicelles by tumor cells, nanomicelles encapsulating DiR, a hydrophobic fluorescent substance, encapsulated in a carrier instead of PTX, were prepared. U87MG cells were seeded at a density of 6 × 104 cells/well in 12-well plates and cultured overnight at 37 ℃, after which the cells were treated with various formulations and cultured for 4 h. The uptake of PEG-PMs and TfR-T12-PMs by U87MG cells was studied by flow cytometry and confocal microscopy, and cellular uptake of the formulations was assessed using DiR as a probe. After treating the cells with different formulations of nanomicelles at 37 ℃ for 6h, uptake of TfR-T12-PMs was significantly higher than PEG-PMs (as shown in FIG. 4 a). This indicates that the presence of TfR-T12 effectively increased PMs uptake by U87MG cells. These results indicate that the presence of TfR-T12 has a significant effect on the uptake capacity of nanomicelles. Confocal microscopy also investigated the distribution of nanomicelles within U87MG cells (as shown in figure 4 b). U87MG cells were treated with various formulations at 37 ℃ for 4h, and the PEG-PMs group showed a weak red intracellular fluorescence, whereas the TfR-T12-PMs group showed enhanced uptake and uptake.
Example 4
CCK8 experiment
U87MG cells were seeded in 96-well plates at 6000 density/well and cultured overnight for fusion. The medium is then removed and nanomicelles or free PTX are added. After 24h incubation, 10 μ L of CCK8 working solution was added and incubated for 1-4h, and the viability per well was calculated. PTX was dose-dependent on the antiproliferative effect of U87MG cells. Cell viability decreased significantly with increasing PTX concentration. All formulations, including free PTX, PEG-PMs and TfR-T12-PMs, showed increased proliferation inhibition of U87MG cells compared to the blank (as shown in FIG. 5). Increased cytotoxicity in these cells is considered to be a promising improvement in the therapeutic effect of PTX, which may be attributed to a combination of higher PTX delivery to cancer cells and intracellular stable release of PTX when it is entrapped in TfR-T12-modified nanocolloid. Thus, the increased PTX toxicity of TfR-T12-PMs was attributable to higher uptake of PTX by U87MG cancer cells. In conclusion, the antiproliferative effect of the preparation on U87MG cells is in the order of TfR-T12-PMs > PEG-PMs > PTX.
Example 5
Construction and evaluation of in vitro blood brain barrier model
A HUVEC/U87MG co-culture model is established. Briefly, 5X 103HUVEC and 2.5X 104U 87MG cells were co-cultured for 72h, HUVEC were seeded into the upper chamber, and U87MG were seeded into the lower chamber. The above inserts were transferred to another 24-well plate, which was pre-seeded with U87MG cells at a concentration of 5 × 104, and the cells were then treated with free DiR and different DiR preparations. After 1h incubation, the upper chamber was removed and the uptake of DiR in U87MG cells was detected by flow cytometry. It is reported that the tumor cells stimulate the endothelial cells of the human umbilical vein and endow the endothelial cells with certain angiogenic properties, so that the human umbilical vein endothelial cells show higher proliferation rate after being cultured with the U87MG cells for 3 days. A human umbilical vein endothelial cell/U87 MG co-culture model is established as a BBTB model to evaluate the transport efficiency of the BBTB cells of the nano-micelles. The results showed (as shown in FIG. 6) that the cell viability of human umbilical vein endothelial cells via TfR-T12-PMs was slightly reduced. After a period of incubation, uptake by TfR-T12-PMs group U87MG cells was significantly higher than PEG-PMs group, and significant apoptosis was also observed in the TfR-T12-PMs group, suggesting that TfR-T12 modified PMs promote transport across the BBTB and targeted tumor cells through the interaction between TfR-T12 peptide and TfR.
Example 6
Construction of subcutaneous brain glioma model and evaluation of nano-micelle tumor inhibition effect
Male nude mice (4-6 weeks) were implanted with 1X 107U87MG cells subcutaneously in the right leg. After 2 weeks, when the solid tumor size reached about 80mm3, the nude mice were randomly divided into 4 groups (n ═ 5) and received intravenous saline, PTX, unmodified nanomicelles, and TfR-T12 modified nanomicelles at a PTX dose of 5 mg/kg. Tumor volume and mouse body weight were measured every 2 days, and tumor volume was calculated according to the following formula (length) × (width) 2/2. The anti-tumor efficacy of nanomicelles is illustrated in fig. 7a and 7b using a xenograft model. Tumor growth was significantly inhibited following continuous intravenous PTX formulation compared to the control group, and in particular, the tumor growth exhibited a more significant reduction in the group treated with TfR-T12-PMs. It demonstrates the superiority of combining TfR-T12 peptide with nano-formulations. In contrast, untreated mice from the saline group showed rapid tumor growth.
Example 7
Experiment for inhibiting angiogenesis
Tumor tissues from tumor-bearing mice were fixed in 4.0% paraformaldehyde solution, embedded in paraffin and sectioned. Immunohistochemistry measures the expression of CD31 in tumor tissue, CD31 being a marker for angiogenesis in tumor tissue. The expression of CD31 in tumor tissues was measured by immunohistochemistry to measure the effect of nanomicelle in inhibiting angiogenesis in tumor tissues, and compared with saline, PTX and PEG-PMs, TfR-T12-PMs significantly inhibited angiogenesis (as shown in FIG. 8).
Example 8
Construction of in-situ brain glioma model and life cycle evaluation
And establishing an in-situ brain glioma model. Briefly, male nude mice (6-8 weeks) were anesthetized with chloral hydrate and approximately 4-5 μ L of 5X 105U87MG cells were implanted into the right brain (2.0 mm parajuxtapose, 3.5 mm depth) using a stereotaxic apparatus (Stoteling). Tumor cells were randomly divided into 4 groups (n ═ 5) 14 days after inoculation, and injected intravenously with saline, PTX, unmodified nanomicelles (PEG-PMs), and TfR-T12 modified nanomicelles (TfR-T12-PMs). The administration is carried out for 5 times in total, the dosage is 5mg/kg, and the survival rate of tumor-bearing mice is observed after 5 times of administration. The survival cycle results of tumor-bearing mice show that the survival rates of the PTX group and the PEG-PMs group are slightly improved compared with those of the normal saline group. However, the median survival for the TfR-T12-PMs group was greater than that for the PEG-PMs group, which was 46 days, and the median survival for the TfR-T12-PMs group was 63 days (as shown in FIG. 9).
Example 9
Evaluation of safety of Nano-micelle
To assess the safety of nanomicelles against tumor-bearing mice, tumor-bearing mice were euthanized after dosing and tissue sections of vital organs were H & E stained for histochemical analysis. Combining the above data, the antitumor activity of TfR-T12-PMs was significantly improved without significant side effects. H & E staining of heart, liver, spleen, lung and kidney was also used for safety assessment. The liver necrosis and lung injury phenomenon occurred in the PTX group, mild heart injury occurred in the PEG-PMs group, and organ injury phenomenon did not occur in the TfR-T12-PMs group (as shown in FIG. 10). When PTX is encapsulated by the vector, its side effects are reduced. Therefore, the TfR-T12 modified nano-micelle is expected to develop a brain tumor targeting preparation, and lays a theoretical foundation for clinical application in the future.
In conclusion, the invention provides a transferrin receptor targeted nano micelle, which comprises TfT-T12 short peptide and polyethylene glycol-polylactic acid connected with the TfT-T12 short peptide through amido bond. The invention utilizes the characteristics of good biological safety, biocompatibility and the like of TfT-T12-PEG-PLA and contains a hydrophilic part and a hydrophobic part, so that the nano-micelle can be formed by self-assembly in water, has stronger transferrin targeting property, and can be used as a drug carrier to carry drugs to cross a blood brain barrier, therefore, the nano-micelle can be used as a transferrin targeting carrier and can be widely applied to the field of treatment of brain glioma.
It is to be understood that the invention is not limited to the examples described above, but that modifications and variations may be effected thereto by those of ordinary skill in the art in light of the foregoing description, and that all such modifications and variations are intended to be within the scope of the invention as defined by the appended claims.
Claims (10)
1. A transferrin receptor targeted nanomicelle, which is characterized by comprising TfT-T12 short peptide and polyethylene glycol-polylactic acid connected with the TfT-T12 short peptide through amido bond.
2. The transferrin receptor targeted nanomicelle according to claim 1, wherein the amino acid structural sequence of the TfT-T12 short peptide is THRPPMWSPVWP.
3. The transferrin receptor targeted nanomicelle according to claim 1, wherein the size of the nanoparticle is 100-200 nm.
4. A method for preparing the transferrin receptor targeted nanomicelle according to claims 1 to 3, comprising the steps of:
dissolving polyethylene glycol-polylactic acid in an organic solvent, adding 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and benzotriazole for activation treatment to obtain an activated polyethylene glycol-polylactic acid solution;
TfT-T12 short peptide and triethylamine are added into the activated polyethylene glycol-polylactic acid solution, and the nano micelle targeted by the transferrin receptor is generated through reaction.
5. The method for preparing the transferrin receptor targeted nanomicelle according to claim 4, wherein the organic solvent is N, N-dimethylformamide or dimethylsulfoxide.
6. The method for preparing transferrin receptor targeted nanomicelle according to claim 4, wherein the activation treatment time is 2-3 h.
7. Use of the transferrin receptor targeted nanomicelle according to any of claims 1 to 3, wherein said transferrin receptor targeted nanomicelle is used to encapsulate a hydrophobic chemotherapeutic agent.
8. The use of the transferrin receptor targeted nanomicelle according to claim 7, wherein the drug loading of the transferrin receptor targeted nanomicelle is between 1% and 10%.
9. The use of the transferrin receptor targeted nanomicelle according to claim 7, wherein the hydrophobic chemotherapeutic agent is paclitaxel.
10. The use of the transferrin receptor targeted nanomicelle according to claim 9, wherein the mass ratio of the transferrin receptor targeted nanomicelle to paclitaxel is 10: 1.
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