CN112933038A - Drug-loaded temperature-sensitive hydrogel delivery system and preparation method and application thereof - Google Patents
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Abstract
The invention discloses a drug-loaded temperature-sensitive hydrogel delivery system and a preparation method and application thereof. The system comprises a temperature-sensitive supramolecular polymer, a drug and a micromolecular polypeptide; the drug is a chemotherapeutic drug; the small molecular polypeptide is an immunologic adjuvant. The preparation method comprises the following steps: the medicine, the small molecular polypeptide and the temperature-sensitive supermolecule polymer are mixed through simple physics, added with sterile phosphate buffer solution, stirred evenly and then placed in an environment with the temperature of more than 30 ℃ to form gel, thus obtaining the gel. After the drug-loaded temperature-sensitive hydrogel delivery system is injected to the periphery of a tumor tissue in a tumor to form colloid, the drug can be continuously and slowly released in the tumor tissue. In a mouse CT26 colorectal cancer subcutaneous tumor model, the drug-loading temperature-sensitive hydrogel drug delivery system has better biological safety, can effectively activate the immune effect of a mouse body and inhibit the growth of tumors, and has huge clinical application potential.
Description
Technical Field
The invention relates to the field of high-molecular drug carriers, in particular to a drug-loading temperature-sensitive hydrogel delivery system and a preparation method and application thereof.
Background
Cancer is one of the factors affecting human life health. Currently, common cancer therapies include: surgical treatment, radiation treatment, chemotherapy, targeted therapy, and the like. Among them, chemotherapy is one of the most commonly used therapies in clinical practice.
Chemotherapy usually enters blood circulation by intravenous injection of chemotherapeutic drugs, and the drugs are transported to tumor tissues through systemic blood circulation, so that tumor cells are killed to realize the purpose of cancer treatment. However, since small molecule chemotherapy drugs have no specific recognition ability for tumor cells after intravenous administration, they can kill normal cells while killing tumor cells, resulting in severe adverse side effects such as severe nausea, vomiting, allergy, renal toxicity, hepatotoxicity, etc. In addition, intravenous injection of small molecule chemotherapy drugs is easily cleared by the reticuloendothelial system (RES) in vivo, resulting in extremely low drug amount reaching tumor tissues and poor treatment effect, and multiple drug resistance of chemotherapy is generated after multiple administrations. In order to solve the above problems, clinical methods for reducing the toxic and side effects of chemotherapeutic drugs, increasing the local drug concentration, and reducing systemic toxicity are always sought, for example: the nano-drug delivery system passively targets tumor tissue enrichment through an EPR effect, and locally injects hydrogel drugs to improve the concentration of the drugs in the tumor tissue, and the like. Among them, the hydrogel has attracted much attention due to its excellent biocompatibility, biodegradability, and bionic properties, and can be retained in tumor tissue as a drug reservoir for a long time and continuously release the drug to increase the drug concentration at the lesion site.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide a drug-loaded temperature-sensitive hydrogel delivery system and a preparation method and application thereof. The drug-loaded temperature-sensitive hydrogel delivery system provided by the invention is injected to a tumor focus and then gels, can be retained for a long time and continuously releases the drug, kills tumor cells and activates the immune effect of an organism, and enhances the treatment effect of tumors.
The purpose of the invention is realized by the following technical scheme.
A drug-loaded temperature-sensitive hydrogel delivery system comprises a temperature-sensitive supramolecular polymer, a drug and a small molecular polypeptide; the drug is a chemotherapeutic drug; the small molecular polypeptide is an immunologic adjuvant.
Preferably, the structure of the temperature-sensitive supramolecular polymer is as follows:
wherein n >0, x >0, y >0, and are integers.
Preferably, the preparation of the temperature-sensitive supramolecular polymer comprises the following steps:
adding a methacrylic anhydride modified hyaluronic acid solution, a double-bond functionalized ureido pyrimidone functional monomer solution and a 2- (2-methoxyethoxy) ethyl methacrylate solution into an oxidation-reduction initiation system of potassium persulfate and tetramethylethylenediamine, and carrying out free radical reaction in a nitrogen atmosphere to obtain a temperature-sensitive supramolecular polymer; the mass ratio of the methacrylic anhydride modified hyaluronic acid, the double-bond functionalized ureidopyrimidone functional monomer to the 2- (2-methoxyethoxy) ethyl methacrylate is (0.05-0.2): (0.01-0.1): (0.15-0.3).
Preferably, the preparation of the temperature-sensitive supramolecular polymer comprises the following steps:
firstly, enabling isocyano ethyl methacrylate (ICEMA) and 2-amino-4-hydroxy-6-Methylpyrimidine (MIS) to generate a rapid coupling reaction in an anhydrous DMSO solvent to prepare a double-bond functionalized ureido-pyrimidone functional monomer UPyMA; preparing a HAMA macromonomer by using sodium Hyaluronate (HA) and Methacrylic Anhydride (MA) under an alkaline condition (pH 8-12); then preparing the temperature-sensitive supramolecular polymer HDU based on free radical reaction between HAMA, UPyMA and 2-methyl-2-acrylic acid-2- (2-methoxy-ethoxy) ethyl ester (DEGMA).
The free radical reaction process of the HDU employs a redox initiation system of potassium persulfate (KPS) and Tetramethylethylenediamine (TEMED).
Preferably, the DMSO is anhydrous DMSO, and the DMSO is stirred in a preheated oil bath at 170 ℃ after being deaerated by introducing nitrogen for 10 min.
The HAMA is prepared in an alkaline environment (pH 8-12), and the temperature is controlled according to the specific reaction stage (the specific steps are that HA is completely dissolved by using deionized water as a solvent and stirred in a low-temperature reactor at 6 ℃ for 12 hours, the temperature is reduced to 0 ℃, MA is dropwise added by using a constant-pressure funnel, 5M NaOH stock solution is dropwise added to maintain the pH between 8-12, the reaction system is changed from transparent to milky white after the NaOH stock solution is added, and the temperature is increased to 4 ℃ for continuous reaction for 12 hours). Since changes in pH and temperature can promote the conversion of MA to methacrylic acid, methacrylic acid cannot react with hydroxyl groups on HA.
Preferably, the chemotherapeutic agent is a chemotherapeutic agent with ICD effect.
Preferably, the chemotherapeutic is one or more of doxorubicin, mitoxantrone, daunorubicin, oxaliplatin and 5-fluorouracil.
Preferably, the immunoadjuvant isDOne or more of PPA-1 (Top-peptide-630184, Shanghai Tanpai Biotech Co., Ltd.), aPD-L1 and aPD-1 immune checkpoint blocking agents.
Preferably, the HDU mass fraction is 4 wt%.
Preferably, the concentration of the drug in the system is 6.25-200. mu.g/100. mu.L.
Preferably, the concentration of the small molecule polypeptide in the system is 4-16mg/100 μ L.
The preparation method of the drug-loaded temperature-sensitive hydrogel delivery system comprises the following steps:
mixing the drug, the small molecular polypeptide and the temperature-sensitive supramolecular polymer through simple physics, adding sterile phosphate buffer solution, stirring uniformly, then placing in an environment with the temperature of more than 30 ℃ for gelling to obtain a drug-loaded temperature-sensitive hydrogel delivery system, which is marked as DOX&DPPA-1@HDU。
Preferably, the stirring temperature is 4 ℃, and the stirring time is 12 hours; the temperature for gelling is 37 ℃ and the time is 1-10 min.
The drug-loaded temperature-sensitive hydrogel delivery system is applied to preparation of tumor treatment drugs.
The temperature-sensitive hydrogel HDU takes HA as a main chain, PDEGMA as a temperature-sensitive chain segment and UPyMA as a quadruple hydrogen bond chain segment, and HAs the advantages that the HA is glycosaminoglycan, and HAs good viscoelasticity and shear thinning properties due to larger molecular weight and hydrophilic repeating units; HA can play a role in regulating cell behaviors in aspects of embryonic development, inflammatory reaction, tumor diffusion and the like, so HA HAs good biocompatibility and bioactivity; ③ the degradation products of HA can promote the process of vascularization; the PDEGMA chain segment enables the HDU to have the characteristic of temperature sensitivity, and the HDU is in a liquid state when the temperature is lower than the LCST and is in a gel state when the temperature is higher than the LCST; the hydrogel is biodegradable, and the final degradation product of the hydrogel does not have adverse effect on organisms; the preparation of the drug-loaded hydrogel is simple, and the tumor drug, HDU and a proper amount of 1xPBS are stirred at 4 ℃ overnight.
After the drug-loaded temperature-sensitive hydrogel delivery system is injected to the periphery of a tumor tissue in a tumor to form colloid, the drug can be continuously and slowly released in the tumor tissue. On one hand, the chemotherapeutic drugs can directly kill tumor cells, cause the cells to generate immunogenic death, release signal molecules such as CRT, HMGB1 and ATP, promote DC maturation and activate the immune effect of mice. On the other hand, the released small molecule polypeptide can be combined with PD-L1 over-expressed on the surface of the tumor, the combination of PD-1 expressed on the surface of a T cell and PD-L1 on the surface of the tumor cell is blocked, the activity of the T cell is maintained, and thus the immune effect is enhanced.
Compared with the prior art, the invention has the following advantages:
the temperature-sensitive hydrogel synthesized by the invention can endow the supermolecule hydrogel with unique physicochemical properties based on the interaction of PDEGMA hydrophobic self-assembly and UPy multiple hydrogen bonds, such as: shear thinning, temperature responsiveness and the like. HDUs also have good biocompatibility and degradability. The drug-loaded temperature-sensitive hydrogel delivery system can realize the long-time sustained release of tumor drugs after directly injecting gel at the focus part, has good safety and huge clinical application potential.
Drawings
FIG. 1 shows the scheme for the synthesis of the supramolecular polymer HDU.
FIG. 2 is a diagram of a supramolecular polymer HDU1H NMR。
FIG. 3 shows drug-loaded temperature-sensitive hydrogel DOX&DThe PPA-1@ HDU co-delivery system is in a gel schematic.
FIG. 4 shows drug-loaded temperature-sensitive hydrogel DOX&DPPA-1@ HDU in vitro drug release profile.
FIG. 5 shows the cytotoxicity test of CT26 on colorectal cancer of mice after DOX is released by drug-loaded temperature-sensitive hydrogel DOX @ HDU.
FIG. 6 is a schematic diagram of calreticulin eversion of CT26 cells of mouse colorectal cancer after DOX is released by detecting drug-loaded temperature-sensitive hydrogel DOX @ HDU through a flow cytometer.
FIG. 7 shows a thermosensitive hydrogel for detecting drug loading by a flow cytometerDPPA-1@ HDU releaseDPPA-1 was followed by binding to cell surface PD-L1 of mouse colorectal cancer CT 26.
FIG. 8 shows that the small animal imager detects the temperature-sensitive hydrogel DOX carrying the drug&DPPA-1@ HDU extended drug retention profiles.
FIG. 9 shows drug-loaded temperature-sensitive hydrogel DOX&DPPA-1@ HDU co-delivery system therapeutic schematic.
Detailed Description
Specific implementations of the present invention are further described below with reference to examples, but the embodiments of the present invention are not limited thereto.
Example 1 Synthesis of temperature-responsive supramolecular Polymer HDU
The synthesis route of HDU is shown in FIG. 1, firstly, double-bond functionalized ureido-pyrimidone group monomer UPyMA is prepared by fast coupling reaction of isocyano ethyl methacrylate (ICEMA) and 2-amino-4-hydroxy-6-Methylpyrimidine (MIS) in an anhydrous DMSO solvent; preparing a HAMA macromonomer by using sodium Hyaluronate (HA) and Methacrylic Anhydride (MA) under an alkaline condition (pH 8-12); then the temperature-sensitive supramolecular polymer HDU is prepared by free radical polymerization based on HAMA, UPyMA and 2-methyl-2-acrylic acid-2- (2-methoxy-ethoxy) ethyl ester (DEGMA).
The detailed synthetic procedure for UPyMA is as follows: weighing 4g of MIS, adding the MIS into a 250mL single-neck round-bottom flask (24#), introducing nitrogen for 10 minutes, removing oxygen, and placing the MIS in a preheated oil bath at 170 ℃; 50mL of anhydrous DMSO is extracted by a 60mL syringe and added, and the mixture is magnetically stirred to be completely dissolved; withdrawing the reaction bottle from the oil bath, placing the reaction bottle on a magnetic stirrer in the air, extracting 5mL of ICEMA by using a syringe under the condition of vigorous stirring, and gradually separating out white precipitate in the cooling (ice bath) process; adding 100mL of acetone, stirring continuously, pouring out the cup, adding 100mL of acetone, stirring continuously, and filtering. The obtained white crude product was repeatedly washed with acetone three times, vacuum-dried at 40 ℃ (yield: 90%), and stored in a refrigerator at 4 ℃ for further use.
The detailed synthesis procedure of HAMA is as follows: weighing 2g of HA, adding 200mL of deionized water, and stirring in a low-temperature reactor at 6 ℃ for 12 hours to completely dissolve the HA; reducing the temperature to 0 ℃, and dropwise adding 15mL of MA into a constant pressure funnel; dropwise adding 5M NaOH stock solution, maintaining the pH value between 8 and 12, and changing the reaction system from transparent to milky white after adding the NaOH stock solution; heating to 4 ℃ and continuing the reaction for 12 hours; precipitating with 1L cold anhydrous ethanol to obtain white floccule, adding deionized water, stirring and dissolving completely, dialyzing in dialysis bag with molecular weight cutoff of 8000-; after the dialysis was finished, the white sponge-like product was obtained by freeze-drying (yield: 85%), and the product was stored in a refrigerator at-20 ℃ for later use.
The detailed synthesis procedure of HDU is as follows: weighing 0.2g of HAMA, adding into a 250mL (24#) single-neck round-bottom flask, adding 180mL of deionized water, and magnetically stirring for 12 hours until the solution is completely dissolved; weighing 0.1g of UPyMA monomer, placing the UPyMA monomer in a 20mL glass bottle, adding 5mL of deionized water, adding 200 mu L of 5M NaOH stock solution, continuing to magnetically stir and completely dissolve the UPyMA monomer, and injecting the UPyMA monomer into a reaction bottle by using an injector; adding 500 mu L (0.5g) of DEGMA into 5mL of deionized water, dissolving completely at low temperature, and adding into a reaction bottle by using an injector; weighing 40mg of KPS, adding 5mL of deionized water, stirring at low temperature to dissolve completely, and injecting into a reaction bottle by using an injector; introducing nitrogen to remove oxygen for 1 hour; adding 200 mu L of TEMED into 5mL of deionized water, deoxidizing, and adding into a reaction bottle; keeping the positive pressure of nitrogen, and reacting for 12 hours at 25 ℃; dialyzing in 8000-12000 dialysis bag for three days, changing water three times per day, lyophilizing to obtain white sponge product (yield: 88%), and storing in 4 deg.C refrigerator.
As shown in fig. 2, by1H NMR characterization shows that the double bond characteristic peak of the reacted monomer is completely proved by comparing the double bond characteristic peak of HAMA at the position of 5-6ppm and the double bond characteristic peak of HD and HDU at the position disappears. The characteristic peak (-COO-CH) of PDEGMA appears at the position of 4.2ppm2-), the 3.22ppm position is attributed to the characteristic peak of the terminal methyl group of PDEGMA, and the characteristic peaks of the other methylene groups on the side chain are located between 3.5 and 4 ppm. The peaks between 3-4ppm overlap with the positions of 10 hydrogens on the glycosidic bond of the disaccharide unit of HA. About 1ppm is hydrogen on the PDEGMA skeleton chain. 1.9ppm (-CH)3) Methyl groups belonging to the HA chain and methylene groups (-CH) on the side chain backbone chain2)。
Example 2 drug-loaded temperature sensitive hydrogel System DOX&DThe preparation method of PPA-1@ HDU and the application I are the establishment of a drug-loaded temperature-sensitive hydrogel system, and the specific method comprises the following steps:
mixing HDU temperature-sensitive supramolecular polymer, chemotherapeutic drug DOX and micromolecular polypeptideDPPA-1 is physically mixed, dissolved by PBS, stirred overnight at 4 ℃, after being uniformly mixed, placed in an environment of 37 ℃ for 1min to observe the change from sol to gel state, as shown in figure 4, the drug-loaded temperature-sensitive hydrogel is injected around the colorectal cancer of a mouse, the hydrogel system changes from sol to gel state at the body temperature of the mouse to be fixed around the tumor, and then DOX and DOX are addedDThe PPA-1 is released from the hydrogel continuously, and the chemotherapeutic drug DOX can kill tumor cells directlyThe cells cause the calreticulin on the surface of tumor cells to turn out, generate ICD effect, release HMGB-1 and ATP, promote DC maturation, activate immune effect of the body, and releaseDPPA-1 can bind with PD-L1 on the surface of tumor cell, maintain T cell activity, kill tumor cell, and enhance the therapeutic effect of tumor.
Second, drug-loaded temperature-sensitive hydrogel system DOX&DIn vitro drug delivery of PPA-1@ HDU
Chemotherapeutic drug DOX and small molecule polypeptideDThe release of PPA-1 was performed in phosphate buffered saline (PBS, pH 7.4) containing 0.02M. Each set of three parallel experiments, 1mL DOX was taken&DPPA-1@ HDU, 2mL of PBS buffer was added. At the specified time point, 1mL of the release solution was removed and supplemented with an equal amount of fresh buffer. Analyzing DOX concentration in the released external liquid by fluorescence spectrophotometer, and analyzing in the drug released liquid by High Performance Liquid Chromatography (HPLC)DThe concentration of PPA-1 is shown in FIG. 4.
In-vitro anti-tumor treatment experiment of drug-loaded temperature-sensitive hydrogel system DOX @ HDU delivery system
1. The DOX has the effect of killing tumors, and the MTT colorimetric method is utilized to detect the proliferation inhibition effect of the prepared drug-loaded temperature-sensitive hydrogel system on the CT26 of the colorectal cancer cells of mice. The specific method comprises the following steps: firstly, 3X 10 of each hole in a 24-hole plate4Individual cells/500. mu.L DMEM medium (containing 10% FBS + 1% diabody), in CO2Incubator (37 ℃, CO)2 Concentration 5%) for 24 h. And preparing different contents of DOX (6.25 mug, 12.5 mug, 25 mug, 50 mug, 100 mug and 200 mug) and equal volume hydrogel in advance, quickly adding the mixed solution into an upper chamber of a Transwell, putting the upper chamber into an empty 24-hole plate, putting the upper chamber into an incubator at 37 ℃ for several minutes until the mixed solution becomes gel, transferring the small chamber of the Transwell into the hole plate containing cells for culture for 12 hours, and culturing the original culture solution for 24 hours. The culture medium was changed and the activity of each group of cells was measured by the MTT method. As can be seen from FIG. 5, the killing effect of DOX on mouse colorectal cancer cell CT26 gradually increased with the increase of the drug concentration.
2. DOX produces ICD effects when killing tumor cells. Evaluation of the above preparation by high-end analytical flow cytometerThe DOX gradually released by the drug-loaded temperature-sensitive hydrogel co-delivery system has the effect of causing the tumor cells to generate immunogenic death. The specific method comprises the following steps: firstly, 1X 10 of each hole is planted in a 24-hole plate5Individual cells/500. mu.L DMEM medium (containing 10% FBS + 1% diabody), in CO2The cells were incubated in an incubator (37 ℃ C., CO2 concentration: 5%) for 24 h. And preparing mixed liquor of the drugs DOX (50 mug, 200 mug and 400 mug) with different contents and the equal volume of hydrogel in advance, quickly adding the mixed liquor into an upper chamber of a Transwell, putting the mixed liquor into an empty 24-hole plate, putting the mixed liquor into an incubator at 37 ℃ for several minutes to gelatinize, putting the mixed liquor into a 24-hole plate with planted cells, and incubating for 12 hours. The medium was aspirated off, the cells were digested, collected, washed with 3% BSA in PBS, incubated with Anti-Calretialin 647 antibody for 30min, washed with 3% BSA in PBS, and finally resuspended in 300 μ L PBS and the MFI detected by flow, as shown in fig. 6.
Fourth, drug-loaded temperature-sensitive hydrogelDPPA-1@ HDU delivery systemDPPA-1 and mouse colorectal cancer cell CT26 surface PD-L1 binding effect experiment
The specific method comprises the following steps: firstly, 1X 10 of each hole is planted in a 24-hole plate5Individual cells/500. mu.L DMEM medium (containing 10% FBS + 1% diabody), in CO2Incubator (37 ℃, CO)2 Concentration 5%) for 24 h. And preparing medicines with different contents in advanceDRapidly adding PPA-1(4mg and 16mg) and hydrogel mixture into upper chamber of Transwell, placing in empty 24-well plate, incubating at 37 deg.C for several minutes until it becomes gel, placing in DMEM medium (containing 10% FBS and 1% double antibody), incubating at 37 deg.C for about 1 hr, and adding PD-L1 antibody to obtain the final productDPPA-1 release solution + PD-L1 antibody mixture. Digesting cells, collecting cells, blocking nonspecific adsorption of cells with 10% FBS-containing PBS solution, and collecting the cellsDThe PPA-1 release + PD-L1 antibody mixture was incubated with the cells at 4 ℃ for 1h, the cells were washed three times with 3% BSA in PBS, and finally the cells were resuspended in 300. mu.L PBS and the MFI was detected by flow-cytometry, as shown in FIG. 7.
Fifthly, carrying medicine thermo-sensitive hydrogel DOX&DPPA-1@ HDU co-delivery system for improving drug retention effect in vivoTest (experiment)
The specific method comprises the following steps: after the Balb/c female mouse is planted with the tumor (CT26) for 6-8 weeks, the tumor volume is grown to 70-80 mm3After that, administration (100. mu.L DOX)&DPPA-1@ HDU and Free DOX&DPPA-1(DOX＝20mg/kg、DPPA-1 ═ 10mg/kg)), a hydrogel is formed at the tumor lesion. At the designated time point, the tumor site of the mouse was photographed using a small animal imager, and the change in fluorescence intensity was analyzed, as shown in fig. 8, it was observed that the drug-loaded hydrogel could prolong the DOX residence time.
Sixthly, carrying medicine thermo-sensitive hydrogel DOX&DPPA-1@ HDU co-delivery system in vivo antitumor therapy experiment
The specific method comprises the following steps: after the Balb/c female mouse is planted with the tumor (CT26) for 6-8 weeks, the tumor volume is grown to 70-80 mm3After that, administration (100. mu.L DOX)&DPPA-1@HDU(DOX＝20mg/kg、DPPA-1 ═ 10mg/kg)), a hydrogel can be formed at the tumor lesion. Tumor growth and body weight were observed every two days, mice were sacrificed after 14 days, dissected and tumor-removed, as shown in figure 9, drug-loaded hydrogel DOX&DThe PPA-1@ HDU group can effectively improve the effect of treating tumors.
Claims (10)
1. A drug-loaded temperature-sensitive hydrogel delivery system is characterized by comprising a temperature-sensitive supramolecular polymer, a drug and micromolecular polypeptide; the drug is a chemotherapeutic drug; the small molecular polypeptide is an immunologic adjuvant.
2. The drug-loaded temperature-sensitive hydrogel delivery system according to claim 1, wherein the temperature-sensitive supramolecular polymer has the following structure:
wherein n >0, x >0, y >0, and are integers.
3. The drug-loaded temperature-sensitive hydrogel delivery system according to claim 2, wherein the preparation of the temperature-sensitive supramolecular polymer comprises the following steps:
adding a methacrylic anhydride modified hyaluronic acid solution, a double-bond functionalized ureido pyrimidone functional monomer solution and a 2- (2-methoxyethoxy) ethyl methacrylate solution into an oxidation-reduction initiation system of potassium persulfate and tetramethylethylenediamine, and carrying out free radical reaction in a nitrogen atmosphere to obtain a temperature-sensitive supramolecular polymer;
the mass ratio of the methacrylic anhydride modified hyaluronic acid, the double-bond functionalized ureidopyrimidone functional monomer to the 2- (2-methoxyethoxy) ethyl methacrylate is (0.05-0.2): (0.01-0.1): (0.15-0.3).
4. The drug-loaded temperature-sensitive hydrogel delivery system according to any one of claims 1 to 3, wherein the chemotherapeutic drug is a chemotherapeutic drug having ICD effect.
5. The drug-loaded temperature-sensitive hydrogel delivery system according to claim 4, wherein the chemotherapeutic drug is one or more of doxorubicin, mitoxantrone, daunorubicin, oxaliplatin and 5-fluorouracil; the immunologic adjuvant isDOne or more of PPA-1, aPD-L1, and aPD-1 immune checkpoint blockers.
6. The drug-loaded temperature-sensitive hydrogel delivery system according to claim 5, wherein the concentration of the drug in the system is 6.25-200 μ g/100 μ L.
7. The drug-loaded temperature-sensitive hydrogel delivery system according to claim 6, wherein the concentration of the small molecule polypeptide in the system is 4-16mg/100 μ L.
8. The method for preparing a drug-loaded temperature-sensitive hydrogel delivery system according to any one of claims 1 to 7, comprising the steps of:
the drug, the small molecular polypeptide and the temperature-sensitive supramolecular polymer are simply and physically mixed, added with sterile phosphate buffer solution, uniformly stirred and then placed in an environment with the temperature of more than 30 ℃ to form gel, so that a drug-loaded temperature-sensitive hydrogel delivery system is obtained.
9. The drug-loaded temperature-sensitive hydrogel delivery system according to claim 8, wherein the stirring temperature is 4 ℃ and the stirring time is 12 h; the temperature for gelling is 37 ℃ and the time is 1-10 min.
10. The use of a drug-loaded temperature-sensitive hydrogel delivery system according to any one of claims 1 to 7 in the preparation of a medicament for the treatment of tumors.
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| CN114426656A (en) * | 2022-01-28 | 2022-05-03 | 青岛科技大学 | Polymer nano hydrogel, drug delivery system comprising same and preparation method thereof |
| CN115317461A (en) * | 2022-08-05 | 2022-11-11 | 常州大学 | Adriamycin delivery system Cu-GA-DOX NPs and preparation method thereof |
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