CN110624113B - Ultrasonic preparation method and application of targeted polyethylene glycol nanoparticle drug carrier - Google Patents
Ultrasonic preparation method and application of targeted polyethylene glycol nanoparticle drug carrier Download PDFInfo
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Abstract
The invention relates to an ultrasonic preparation method and application of a targeted polyethylene glycol nanoparticle drug carrier. Meanwhile, the ultrasonic polymerization method also has the advantages of short polymerization time and high monomer conversion efficiency, is used for loading the antitumor drugs to obtain the targeted polyethylene glycol nano-drug carrier (Pt-loaded PEG-RGD NPs) loaded with the cisplatin prodrug molecules, can realize the controllable release of the anticancer drug molecules cisplatin based on the characteristics of a tumor microenvironment, and reduces the toxic and side effects of the anticancer drug molecules.
Description
Technical Field
The invention relates to an ultrasonic preparation method and application of a targeted polyethylene glycol nanoparticle drug carrier, belonging to the technical field of high polymer materials.
Background
Cancer is a worldwide scientific problem in the 21 st century and is also one of the diseases which seriously affect the human life and health in the world today. The common treatment means includes operation treatment, radiotherapy, chemotherapy, gene treatment, immunotherapy and the like. Chemotherapy is an important cancer treatment means, and has very good application in the aspects of tumor control, postoperative treatment and the like. However, the traditional anticancer drugs have the defects of low drug utilization rate, large toxic and side effects, no specific identification and the like. In recent years, different types of drug carriers (e.g., micelles, capsules, dendrimers, inorganic nanoparticles, proteins, hydrogels, etc.) have been developed. The design of these drug carriers only alleviates the problems of low utilization rate and large toxic and side effects to some extent, but does not solve the problems fundamentally. Therefore, how to enhance the utilization rate and safety of chemical drugs has irreplaceable significance for improving the treatment effect of tumors and human health, and is also a major challenge in current scientific and clinical research.
Prolonging the circulation time of the carrier in blood and increasing the probability of enrichment at tumor sites are effective ways to improve the transmission efficiency of anticancer drugs. Polyethylene glycol (Poly) is a polymeric material approved by the U.S. food and drug safety administration (FDA) for use in biopharmaceuticals. Since the first report in 1977 that the nano-drug carrier can be used for prolonging the circulation time in vivo, the nano-drug carrier has been widely used for surface modification. The polyethylene glycol can reduce protein in nanometer level due to its high hydration propertyThe adsorption on the surface of the particle is favorable for reducing the nonspecific interaction between the carrier and organisms and prolonging the circulation time in vivo. Currently, some polyethylene glycol surface-modified nano drug carriers (such as:
and
) Has been marketed and has achieved effective therapeutic effects in the clinical field. Self-assembling polyethylene glycol-containing block polymers and post-modifying polyethylene glycols are two major pegylation strategies. However, the steric hindrance of polyethylene glycol is large, and the modification density of polyethylene glycol is low, so that the biological antifouling performance is poor. In recent years, nanocarriers mainly comprising polyethylene glycol as a building element have been designed and reported, which overcome the problem of insufficient density of polyethylene glycol modification to some extent. However, polyethylene glycol nanoparticles can reduce the interaction with tumor cells while interacting non-specifically with organisms, so that the therapeutic effect is reduced. How to increase the targeted delivery efficiency of pure polyethylene glycol drug carriers at tumor sites is an important scientific problem.
At present, the preparation method of the polyethylene glycol nano particles mainly comprises a template method and a bulk polymerization method. For the template method, silica, calcium carbonate, polystyrene and the like are commonly used as templates. The silicon dioxide template method has various shapes and is easy to control the size, but dangerous substances such as hydrofluoric acid and the like are needed to be used for removing the template. Calcium carbonate and polystyrene templates are relatively easy to remove, but are difficult to use to prepare polyethylene glycol particles of small size. For bulk polymerization, the synthesis conditions are usually complex and harsh, time and labor are consumed, and organic substances such as catalysts and the like are generally introduced in the reaction process, so that the bulk polymerization has certain biological toxicity for subsequent application.
Therefore, the development of a novel targeted polyethylene glycol nano-drug carrier is beneficial to improving the preparation efficiency of the nano-carrier, reducing the nonspecific interaction between the carrier and an organism, improving the targeted transmission efficiency of the carrier and enhancing the tumor treatment effect.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides an ultrasonic preparation method and application of a targeted polyethylene glycol nanoparticle drug carrier.
In the specification, 2-aminoethyl methacrylate hydrochloride is abbreviated as: AEMA;
4- (4, 6-dimethoxytriazine) -4-methylmorpholine hydrochloride, abbreviated as: DMTMM;
methoxy polyethylene glycol methacrylate, polyethylene glycol molecular weight 2k, abbreviation: ACLT-PEG 2K ；
Methoxy polyethylene glycol methacrylate, polyethylene glycol molecular weight 5k, abbreviation: ACLT-PEG 5K ；
The molecular weight of the acrylate polyethylene glycol active ester is 5k, and the molecular weight is as follows: ACLT-PEG 5K -NHS；
The molecular weight of the acrylate polyethylene glycol cyclic RGD is 5k, and the short is: ACLT-PEG 5K -RGD；
Cyclic RGD, abbreviation: CRGDfk;
polyethylene glycol nanoparticles, for short: PEG NPs;
targeting polyethylene glycol nanoparticles, abbreviated: PEG-RGD NPs;
the fluorescent dye AF647 marked polyethylene glycol nano-drug carrier is abbreviated as: AF647-loaded PEG NPs;
the fluorescent dye AF647 marked targeting polyethylene glycol nano-drug carrier is abbreviated as: AF647-loaded PEG-RGD NPs;
loaded with cisplatin prodrug molecule c, c, t- [ Pt (NH) 3 ) 2 Cl 2 (OH)(O 2 CCH 2 CH 2 CO 2 H)]The polyethylene glycol nano-drug carrier is abbreviated as: pt-loaded PEG NPs;
loaded with cisplatin prodrug molecule c, c, t- [ Pt (NH) 3 ) 2 Cl 2 (OH)(O 2 CCH 2 CH 2 CO 2 H)]The targeted polyethylene glycol nano-drug carrier is abbreviated as: pt-loaded PEG-RGD NPs;
hereinafter, the term "a" or "an" is used for brevity.
In order to solve the problems, the invention is realized by the following technical scheme:
a targeted polyethylene glycol nanoparticle drug carrier has a structure shown in the following formula I:
an ultrasonic polymerization preparation method of a targeted polyethylene glycol nanoparticle drug carrier comprises the following steps:
respectively mixing ACLT-PEG, AEMA and ACLT-PEG 5k Adding RGD into ultrapure water, stirring and dissolving to obtain a mixed solution, removing oxygen from the mixed solution, performing ultrasonic polymerization at 35-50 ℃ for 10-30min, wherein the ultrasonic power is 35-45W and the ultrasonic frequency is 400-420kHz, and after the polymerization reaction is finished, purifying, and freeze-drying to obtain the targeted polyethylene glycol nanoparticle drug carrier.
According to the invention, the ACLT-PEG is preferably ACLT-PEG 2k Or ACLT-PEG 5k Preferably, the ACLT-PEG is ACLT-PEG 2k 。
According to the invention, preferably, the ACLT-PEG 5k RGD is prepared as follows: mixing with ACLT-PEG 5k mixing-NHS and cRGDfk according to a molar ratio of 1 5k -RGD。
ACLT-PEG 5k Preparation of RGD as shown in scheme 1 below:
according to a preferred embodiment of the invention, the ratio of ACLT-PEG: AEMA: ACLT-PEG 5k -the molar ratio of RGD is: (5-7): (2-4): (0.5-1.5).
Further preferably, the ratio of ACLT-PEG: AEMA: ACLT-PEG 5k -the molar ratio of RGD is: 6:3:1.
According to the invention, preferably, ACLT-PEG, AEMA, ACLT-PEG 5k RGD is added into water, and the total mass concentration of the RGD, the RGD and the water is 15-25%.
According to the invention, the ultrasonic polymerization temperature is 40 ℃, the ultrasonic power is 40W, the ultrasonic frequency is 412kHz, and the ultrasonic time is 20min.
According to the invention, the oxygen removal is preferably performed for 30min by blowing nitrogen.
According to the invention, the polymerization is terminated by using cold water, and the purification is carried out for 2 days by using a dialysis bag of 8k-14k Da.
A targeting polyethylene glycol nanoparticle drug carrier is used for loading antitumor drugs.
A drug-carrying system comprises a targeting polyethylene glycol nanoparticle drug carrier and a drug carried by the targeting polyethylene glycol nanoparticle drug carrier.
According to the invention, the drug loaded on the targeted polyethylene glycol nanoparticle drug carrier is preferably a reduction-response anticancer drug molecule, and the drug loaded on the targeted polyethylene glycol nanoparticle drug carrier is preferably a cisplatin prodrug molecule c, c, t- [ Pt (NH) 3 ) 2 Cl 2 (OH)(O 2 CCH 2 CH 2 CO 2 H)]。
The preparation method of the medicine carrying system comprises the following steps:
adding a targeted polyethylene glycol nanoparticle drug carrier and cisplatin prodrug molecules into a phosphate buffer solution, adding DMTMM as a carboxyl activator, reacting for 24 hours at 25 ℃, and purifying to obtain a targeted polyethylene glycol nanoparticle drug carrier (Pt-loaded PEG-RGD NPs) loading the cisplatin prodrug molecules; the mass ratio of the targeted polyethylene glycol nanoparticle drug carrier to the cisplatin prodrug molecule is as follows: (2-4): (1-3).
Preparation of Pt-loaded PEG-RGD NPs is shown in scheme 2 below:
preferably, according to the present invention, the molar ratio of DMTMM added to cisplatin prodrug molecule is: (3-1): 1.
preferably, according to the invention, the phosphate buffer solution has a pH of 7.4 and a concentration of 10mM, and the mass-to-volume ratio of the targeted polyethylene glycol nanoparticle drug carrier to the phosphate buffer solution is 30.
According to the invention, the purification is preferably carried out for 2 days by dialysis with a dialysis bag of 8k-14k Da.
Preferably, according to the present invention, the cisplatin prodrug molecule is prepared as follows:
(1) Mixing cisplatin with H 2 O 2 Mixing according to the molar ratio of 1 3 ) 2 Cl 2 (OH) 2 ]；
(2) Mixing c, c, t- [ Pt (NH) ] 3 ) 2 Cl 2 (OH) 2 ]Mixing with succinic anhydride according to the molar ratio of 1.
According to the invention, the purification of the step (1) is that the precipitate obtained after the reaction is washed with cold water for 3 times, ethanol for 3 times, ether for 3 times and dried in vacuum.
According to the invention, the purification of step (2) is performed by DMSO freeze drying, adding acetone to precipitate the product, washing with acetone for 3 times, washing with ether for 3 times, and drying.
The cisplatin prodrug molecule was prepared as shown in scheme 3 below:
the preparation method of the targeted polyethylene glycol nanoparticle drug carrier based on the ultrasonic polymerization method has the following advantages:
1. the invention adopts ultrasonic polymerization to prepare targeted polyethylene glycol nanoparticles, and the method has no initiator and catalyst introduction, and is particularly suitable for preparation of biological samples. Meanwhile, the ultrasonic polymerization method also has the advantages of short polymerization time and high monomer conversion efficiency.
2. The targeting polyethylene glycol nano particles prepared by the invention can be freeze-dried into powder for storage, and have good redispersibility. The method has important significance for the storage and transmission of the nano-drugs.
3. The cisplatin-loaded targeted polyethylene glycol nanoparticle prepared by the invention can realize the controllable release of anticancer drug molecules cisplatin based on the characteristics of a tumor microenvironment, and reduce the toxic and side effects of the anticancer drug molecules.
4. The targeting polyethylene glycol nanoparticles prepared by the invention have better biological antifouling performance, and can effectively reduce the nonspecific interaction between a carrier and organisms. The introduction of the targeting molecule effectively improves the specific interaction between the vector and the U87 MG cell. Meanwhile, the introduction of the targeting molecules does not influence the biological antifouling performance of the carrier.
Drawings
FIG. 1 shows the cisplatin prodrug molecule c, c, t- [ Pt (NH) ] 3 ) 2 Cl 2 (OH)(O 2 CCH 2 CH 2 CO 2 H)]Is 1 H nuclear magnetic resonance spectrogram.
FIG. 2 shows ACLT-PEG 5k Of RGD 1 H nuclear magnetic resonance spectrogram.
FIG. 3 shows the Zeta potentials of different support systems.
FIG. 4 is a TEM image of the targeted polyethylene glycol nanoparticle PEG-RGD NPs of example 2 and the drug-loaded PEG-RGD NPs of example 3, wherein a is a TEM image of the Pt-loaded PEG-RGD NPs, and b is a TEM image of the PEG-RGD NPs.
FIG. 5 is an AFM image and a height statistical image of the targeted polyethylene glycol nanoparticle PEG-RGD NPs of example 2 and the Pt-loaded PEG-RGD NPs of example 3; a is an AFM diagram of the Pt-loaded PEG-RGD NPs, b is a high statistical diagram of the Pt-loaded PEG-RGD NPs, c is an AFM diagram of the PEG-RGD NPs, and d is a high statistical diagram of the PEG-RGD NPs.
Fig. 6 is a photograph showing redispersion of the drug-loaded PEG-RGD NPs of example 3, wherein the Pt-loaded PEG-RGD NPs are powdery photographs, a photograph showing dispersion after dissolution in water, and a photograph showing tyndall phenomenon in sequence from left to right, and it can be seen from the photograph showing tyndall phenomenon that the dissolved polyethylene glycol nanocarrier is a stable colloidal dispersion system.
FIG. 7 is the drug release profile of Pt-loaded PEG-RGD NPs in different systems.
FIG. 8 shows PEG 2k NPs、PEG 2k -RGD NPs interaction with RAW 264.7 cells.
FIG. 9 is PEG 2k NPs、PEG 2k Results of interaction of RGD NPs with U87 MG cells.
FIG. 10 is a graph showing cytotoxicity results.
FIG. 11 is a graph showing the results of apoptosis, where a is a blank control group, b is a cisplatin group, c is a cisplatin prodrug group, d is PEG NPs, e is Pt-loaded PEG NPs, and f is Pt-loaded PEG-RGD NPs.
FIG. 12 is PEG 5k NPs、PEG 5k -RGD NPs interaction with U87 MG cells.
The specific implementation mode is as follows:
for a better understanding of the present invention, reference is made to the following detailed description and accompanying drawings.
Example 1
Cisplatin prodrug molecule c, c, t- [ Pt (NH) 3 ) 2 Cl 2 (OH)(O 2 CCH 2 CH 2 CO 2 H)]The synthesis of (a) is carried out,
(1) Mixing cisplatin with H 2 O 2 Mixing according to a molar ratio of 1 3 ) 2 Cl 2 (OH) 2 ]；
(2) Mixing c, c, t- [ Pt (NH) 3 ) 2 Cl 2 (OH) 2 ]Mixing with succinic anhydride according to a molar ratio of 1;
the prepared cisplatin prodrug molecule c, c, t- [ Pt (NH) 3 ) 2 Cl 2 (OH)(O 2 CCH 2 CH 2 CO 2 H)]Is/are as follows 1 The H NMR spectrum is shown in FIG. 1.
ACLT-PEG 5k RGD is prepared as follows: mixing with ACLT-PEG 5k mixing-NHS and cRGDfk according to a molar ratio of 1 5k -RGD。
Prepared ACLT-PEG 5k Of RGD 1 The H NMR spectrum is shown in FIG. 2.
Example 2
Preparing targeted polyethylene glycol nanoparticles, wherein the molecular weight of polyethylene glycol is 2k, and the steps are as follows:
ACLT-PEG 2k (138mg, 0.069mmol), AEMA (4.5mg, 0.0345mmol) and ACLT-PEG 5k RGD (57.5mg, 0.0115mmol) was dissolved in 1mL of ultrapure water, and oxygen was removed by bubbling nitrogen for 30min. The reaction system was subjected to ultrasonic polymerization at 40 ℃ for 20min under 40W and 412kHz. The polymerization was terminated by adding 9mL of cold water to the reaction system. Transferring the reaction solution into a dialysis bag for dialysis for 2 days, and freeze-drying into powder to obtain the targeted polyethylene glycol nanoparticles.
The Zeta potential, the TEM image and the AFM image of the obtained targeting polyethylene glycol nanoparticle are respectively shown in figures 3, 4 and 5.
GPC measurements showed that the molecular weight of the nanoparticles was around 1300kDa, and the hydration kinetic diameter was around 70nm. And further characterizing the morphology of the targeted polyethylene glycol nanoparticles by adopting a TEM and an AFM. The result shows that the microscopic morphology of the targeted polyethylene glycol nanoparticle is spherical, the sizes are different from 30 to 100nm, and the height is 2 to 3nm, which indicates that the nanoparticle is a substance nanoparticle.
Example 3
The preparation of the drug-carrying system comprises the following steps:
20mg (0.046 mmol) of the cisplatin prodrug molecule, 19.09mg (0.069 mmol) of DMTMM and 30mg of the polyethylene glycol nanoparticle PEG NPs of example 2, the polyethylene glycol molecular weight of which is 2k, were weighed and dissolved in 1mL of PBS (10 mM, pH 7.4) buffer solution, and the reaction was stirred at room temperature for 24 hours. After the reaction, the reaction solution was dialyzed for 2 days to remove unreacted cisplatin prodrug molecule and DMTMM as a catalyst. Freeze-drying into powder to obtain the target polyethylene glycol nano-drug carrier (Pt-loaded PEG-RGD NPs) loaded with cisplatin prodrug molecules.
Application example 1:
1. in vitro release of drug molecules
2mL of Pt-loaded PEG-RGD NPs (1 mg mL) were taken -1 ) Placed in an 8k-14k dialysis bag, and the bag was placed in 48mL of PBS buffer solution (10 mM, pH 6.5) containing 5mM sodium ascorbate. 1mL of dialysate was removed at a set time point each time, and 1mL of fresh buffer solution was added. After the end of spotting, the released cisplatin was quantitatively analyzed by ICP-MS to draw a cisplatin release curve. The control group was also prepared by adding dialysate without ascorbic acid, and the release profile of cisplatin prodrug molecule is shown in fig. 7.
The cisplatin prodrug molecule is a drug molecule which is released in a reduction response, and about 80 percent of the cisplatin drug molecule is released after 24 hours in the presence of ascorbic acid. In the control group without ascorbic acid, less than 20% of the cisplatin drug molecules were released after 24h. The reduction response release behavior of the drug molecules was demonstrated.
2. Cell interaction assay
Firstly, RAW 264.7 phagocytes are selected to study the interaction behavior of the nanoparticles and cells. 24-well plates, 50000 cells per well, cultured overnight, cells attached, to the wells added AF647-loaded PEG NPs and AF647-loaded PEG-RGD NPs. The material concentrations were set to 100. Mu.g mL respectively -1 Three groups, the culture time is set to four groups of 2,4,8 and 12 h. After the interaction was completed, the nanoparticles without interaction were washed away with DPBS, cells were digested with pancreatin and collected, dispersed in DPBS solution, and analyzed by flow cytometry, and the results of the detection are shown in fig. 8. The nano particles prepared by the method have better biological antifouling performance, and the introduction of the targeting molecules does not influence the biological antifouling performance.
Secondly, the expression alpha is selected v β 3 Integrin U87 MG cells studied their interaction behavior with nanoparticles. Similarly, U87 MG cells were seeded in 24-well plates at 50000 cells/well, cultured overnight, cells attached to the wall, and AF647-loaded PEG NPs and AF647-loaded PEG-RGD NPs were added. The concentration of nanoparticles was set at 100. Mu.g mL -1 Three groups, the culture time is set to four groups of 2,4,8 and 12 h. After the interaction is finished, washing off the nano particles without the action, digesting by pancreatin, collecting cells, and carrying out detection and analysis by a flow cytometer. The detection result is shown in fig. 9, and the result shows that the nanoparticles modified with the targeting molecule RGD have stronger cell interaction with U87 MG cells, while less than 20% of the cells of the nanoparticles without modified targeting molecule have interaction with the nanoparticles. The prepared nano particles are proved to have better U87 MG cell targeting.
3. Cytotoxicity test
8000 cells per well of 96-well plates were plated, cultured overnight, and cells attached. Cisplatin concentration was set at 0.3,0.6,1.3,2.5,5.0, 10.0. Mu.g mL -1 Six groups, the culture time is 48h. After completion of the action, 10. Mu.L of MTT (5 mg mL) was added to each well -1 ) After 4h of action, formazan was dissolved by adding 100. Mu.L of DMSO. The absorbance was measured by a microplate reader, and the excitation wavelength was 570nm. The results are shown in fig. 10, and from the result of cytotoxicity, the targeted polyethylene glycol nanoparticle loaded with cisplatin drug molecules has greater killing performance on tumor cells, while the targeted group without cisplatin drug molecules has relatively weaker cytotoxicity.
4. Apoptosis assay
50000 cells were seeded per well in 24-well plates, cultured overnight, and cells attached. Drug concentration was set to 5. Mu.g mL -1 Incubation time was 48h, then medium was removed and 500. Mu.L of dye-containing medium, 2. Mu. Mol L per well -1 Calcein-Am and 4. Mu. Mol L of -1 PI of (4). After staining for 10min, observation was performed with a fluorescence microscope, and the results are shown in FIG. 11, where red represents apoptotic cells and green represents viable cells. The result is consistent with the cytotoxicity result, and the apoptosis of the targeted polyethylene glycol nanoparticle group is the most.
Example 4
Preparing targeted polyethylene glycol nanoparticles, wherein the molecular weight of polyethylene glycol is 5k, and the steps are as follows:
ACLT-PEG 5k (171mg, 0.034mmol), AEMA (2.85mg, 0.117mmol) and ACLT-PEG 5k RGD (29mg, 0.0057mmol) was dissolved in 1mL of ultrapure water and purged with nitrogen for 30min to remove oxygen. The reaction system is put under the conditions of 40 ℃,40W and 412kHz for ultrasonic polymerization for 15min. The polymerization was terminated by adding 9mL of cold water to the reaction system. Transferring the reaction solution into a dialysis bag for dialysis for 2 days, and freeze-drying into powder to obtain the targeted polyethylene glycol nanoparticles.
Application example 2: cell interaction assay
Selecting expression alpha v β 3 Integrin U87 MG cells studied their interaction behavior with nanoparticles. Similarly, U87 MG cells were seeded in a 24-well plate at 50000 cells/well, cultured overnight, cells were attached to the wall, and AF647-loaded PEG NPs and AF647-loaded PEG-RGD NPs were added to the wells. The concentration of the nanoparticles was 50, 100, 200. Mu.g mL -1 Three groups, the culture time is set to be four groups of 2,4,8 and 12 h. After the interaction is finished, washing off non-functional nano particles, digesting by using pancreatin, collecting cells, and carrying out detection and analysis by using a flow cytometer. The detection results are shown in fig. 12, and the results show that the interaction degree between the nanoparticles modified with the targeting molecule RGD and the nanoparticles without the modified targeting molecule and the cells is low. This shows that the targeting molecules of RGD are shielded by the longer polyethylene glycol segments, and the targeting property of the polyethylene glycol nanoparticles with the main body building unit of 2k is not shielded.
Claims (5)
1. A preparation method of a medicine carrying system comprises the following steps:
adding a targeted polyethylene glycol nanoparticle drug carrier and cisplatin prodrug molecules into a phosphate buffer solution, adding DMTMM as a carboxyl activator, reacting for 24 hours at 25 ℃, and purifying to obtain a targeted polyethylene glycol nanoparticle drug carrier (Pt-loaded PEG-RGD NPs) loading the cisplatin prodrug molecules; the mass ratio of the targeted polyethylene glycol nanoparticle drug carrier to the cisplatin prodrug molecule is as follows: (2-4): (1-3);
the targeted polyethylene glycol nanoparticle drug carrier is prepared by the following method:
separately mixing ACLT-PEG 2k 、AEMA、ACLT-PEG 5k Adding the-RGD into ultrapure water, stirring and dissolving, and adding ACLT-PEG 2k ：AEMA：ACLT-PEG 5k -the molar ratio of RGD is: 3:1, obtaining a mixed solution, removing oxygen from the mixed solution, performing ultrasonic polymerization at 35-50 ℃ for 10-30min, wherein the ultrasonic power is 35-45W, the ultrasonic frequency is 400-420kHz, and after the polymerization reaction is finished, purifying, and freeze-drying to obtain the targeted polyethylene glycol nanoparticle drug carrier;
the cisplatin prodrug molecule is prepared by the following method:
(1) Mixing cisplatin with H 2 O 2 Mixing according to the molar ratio of 1c,c,t-[Pt(NH 3 ) 2 Cl 2 (OH) 2 ]；
(2) Will be provided withc,c,t-[Pt(NH 3 ) 2 Cl 2 (OH) 2 ]Mixing with succinic anhydride according to the molar ratio of 1.
2. The method of claim 1, wherein the ACLT-PEG is present in the composition 5k -RGD is prepared as follows: mixing with ACLT-PEG 5k mixing-NHS and cRGDFk according to the molar ratio of 1 5k -RGD。
3. The preparation method according to claim 1, wherein the ultrasonic polymerization temperature is 40 ℃, the ultrasonic power is 40W, the ultrasonic frequency is 412kHz, the ultrasonic time is 20min, the oxygen removal is performed by blowing nitrogen for 30min, the polymerization reaction is terminated by using cold water, and the purification is performed by dialyzing for 2 days by using a dialysis bag of 8k-14k Da.
4. The method according to claim 1, wherein the mass ratio of DMTMM to cisplatin prodrug molecule is: (0.8-1): 1, the pH of the phosphate buffer solution is 7.4, the concentration is 10mM, and the mass-to-volume ratio of the targeted polyethylene glycol nanoparticle drug carrier to the phosphate buffer solution is as follows: 30, unit, mg/mL, said purification being dialysis for 2 days using dialysis bags of 8k-14k Da.
5. The drug delivery system of any of claims 1-4, comprising a targeted polyethylene glycol nanoparticle drug carrier and a drug carried by the targeted polyethylene glycol nanoparticle drug carrier; targeting polyethylene glycol nanoparticle drug carrier loaded drug is cisplatin prodrug moleculec,c,t-[Pt(NH 3 ) 2 Cl 2 (OH)(O 2 CCH 2 CH 2 CO 2 H)]。
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