CN111228507B - Gold nanorod drug loading system modified by HPMA polymer and preparation method and application thereof - Google Patents
Gold nanorod drug loading system modified by HPMA polymer and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a gold nanorod drug loading system modified by HPMA polymer, which takes pH sensitive HPMA polymer as a modified chain of a gold nanorod, and the pH sensitive HPMA polymer is connected with chemotherapeutic drugs through pH sensitive chemical bonds. The gold nanorod drug loading system modified by the HPMA polymer utilizes the water-soluble HPMA polymer to assemble chemotherapeutic drugs into GNRs, and acts on prostate cancer cells to improve the killing power of the chemotherapeutic drugs under the synergistic effect of thermotherapy and chemotherapy. The invention also provides a preparation method of the gold nanorod drug delivery system modified by the HPMA polymer, and the preparation process is simple and easy to operate. The invention also provides application of the gold nanorod drug delivery system modified by the HPMA polymer in treating prostate cancer, effectively overcomes drug resistance of tumor cells, and delays occurrence and metastasis of prostate cancer.
Description
Technical Field
The invention relates to the field of pharmaceutical preparations, in particular to a gold nanorod drug delivery system modified by HPMA (high Performance polyethylene) polymer and a preparation method and application thereof.
Background
Prostate cancer is the most common malignant tumor in men in developed countries and regions such as europe and america, and the mortality rate of prostate cancer is the second place of various cancers. The incidence of disease is lower in asia than in western countries, but in recent years there is a rapidly rising trend. When patients diagnose metastatic prostate cancer in early stage, the surgical and endocrine treatment means can temporarily and effectively inhibit the metastasis of soft tissue focus and reduce the level of prostate specific antigen in serum. However, almost all prostate cancer patients inevitably develop castration-resistant prostate cancer (mCRPC), which is not sensitive to endocrine therapy, and only chemotherapy is tried. Among them, docetaxel can significantly improve clinical symptoms of mCRPC bone metastasis patients, but its drug resistance is one of the most common causes of clinical therapeutic ineffectiveness. Unfortunately, the resistance mechanisms of different chemotherapeutic molecules are also different. Therefore, it is necessary to research a novel targeted drug delivery system which is not toxic to normal prostate epithelial cells, can delay the occurrence and metastasis of prostate cancer and overcome multidrug resistance.
In recent years, chemo-photothermal combination therapy (chemo-photothermal combination therapy) has provided a new strategy for drug-resistant cancer therapy. A large amount of research data prove that chemotherapeutic drugs and photothermal reagents are co-transmitted through a nano system and are combined with near-infrared (NIR) laser irradiation, so that the photothermal reagents generate cancer cell sensitive heat, the residence time of the chemotherapeutic drugs in tumors is effectively prolonged, the drugs are promoted to play a role, and multi-drug resistance is overcome. Researchers of Chinese academy of sciences prepare nanoparticles (DINPs) which carry chemotherapy drug adriamycin and photothermal reagent indocyanine green together by a one-step ultrasonic method, and show more obvious induction rate of apoptosis and necrosis and tumor growth inhibition activity on drug-resistant MCF-7/ADR breast cancer cells through the synergistic effect of chemotherapy and thermotherapy. Therefore, the development of a safe and efficient delivery system ensures that the chemotherapeutic drug and the photothermal agent can be simultaneously delivered to the tumor part to play multiple synergistic functions, and overcomes the drug resistance of tumor cells, which is imperative.
Gold Nanorods (GNRs) are used as a novel nano-carrier, and have extremely wide application prospect in the field of biomedicine. It has the advantages of low toxicity, large specific surface area and easy combination with biological molecules, and can be used for transferring micromolecular drugs and biological macromolecules. The near infrared absorption performance of the GNRs can be adjusted by controlling the appearance, size and structure of the GNRs in the synthesis process. However, due to self-aggregation of GNRs in different cells, the ideal near-infrared window may be shifted into the visible region, greatly reducing their photothermal conversion efficiency. In addition, the non-specific transport of GNRs is likely to cause systemic toxic and side effects, and is a problem to be solved at present.
The water-soluble N- (2-Hydroxypropyl) methacrylamide [ N- (2-Hydroxypropyl) -methacrylamide, HPMA ] is used as a synthetic polymer material, has the characteristics of good biocompatibility, no immunogenicity, capability of modifying the structure according to the use purpose and the like. Various HPMA polymer drug conjugates of the linear, graft, and micelle types have been developed that link various drugs with different spacers.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide an HPMA polymer modified gold nanorod drug loading system, which utilizes a water-soluble HPMA polymer to assemble chemotherapeutic drugs into GNRs (GNRs) to cooperate with the thermotherapy chemotherapy effect, thereby solving the drug resistance problem of cells in the prostate cancer treatment process.
The invention also aims to provide a preparation method of the gold nanorod drug-loading system modified by the HPMA polymer.
The invention also aims to provide application of the gold nanorod drug delivery system modified by the HPMA polymer in treating prostate cancer.
One of the purposes of the invention is realized by adopting the following technical scheme:
the drug delivery system takes a pH sensitive HPMA polymer as a modification chain of a gold nanorod, and the pH sensitive HPMA polymer is connected with a chemotherapeutic drug through a pH sensitive chemical bond.
Further, the pH sensitive chemical bond is a hydrazone bond.
Further, the chemotherapeutic drug is doxorubicin.
The second purpose of the invention is realized by adopting the following technical scheme:
the preparation method of the gold nanorod drug delivery system modified by the HPMA polymer comprises the following steps:
(1) preparing CTAB-GNRs by adopting a seed solution growth method;
(2) taking MA-GG-NHNH2Reacting with chemotherapeutic drug adriamycin to obtain intermediate MA-GG-NHN ═ DOX;
(3) preparing a pH-sensitive HPMA polymer PDS-pHPMA-DOX by using HPMA and the intermediate MA-GG-NHN ═ DOX obtained in the step (2) as monomers;
(4) and (3) mixing the CTAB-GNRs prepared in the step (1) with the product PDS-pHPMA-DOX prepared in the step (3), and reacting in a dark place to obtain the gold nanorod drug delivery system pHPMA-DOX @ GNRs modified by the HPMA polymer.
Further, the preparation process of MA-GG-NHN ═ DOX in the step (2) above is: taking MA-GG-NHNH2Dissolving adriamycin in methanol, adding glacial acetic acid, and stirring at room temperature to obtain the product.
Further, the MA-GG-NHNH2And doxorubicin at a 1:1 molar ratio.
Further, the step (3) includes the following steps: placing ABIK-TT, DMSO, monomer HPMA, MA-GG-NHN ═ DOX in an ampoule bottle by taking 4, 4' -azobis (cyanovaleric acid) -thiazolidine-2-thione (ABIK-TT) as an initiator and DMSO as a reaction solvent, sealing, carrying out oil bath reaction at 50 ℃ for 24 hours, dissolving the crude product with ultrapure water, dialyzing for 48 hours, and carrying out freeze drying to obtain a semi-shaking claw polymer TT-pHPMA-DOX with the terminal chain being TT; TT-pHPMA-DOX and PDEA are dissolved in DMF, DMF containing DIEA is dripped into reaction liquid, the mixture is stirred for 3 hours at room temperature, the solvent is removed under reduced pressure, deionized water is used for dissolving the crude product, dialysis is carried out for 48 hours, and the polymer PDS-pHPMA-DOX is obtained after freeze drying.
Further, the weight percentage of the mixture of AIBK-TT, DMSO, monomer HPMA, and MA-GG-NHN ═ DOX is 2.4%, 85.1%, 12.5%, respectively, wherein the molar ratio of monomer HPMA and MA-GG-NHN ═ DOX is 9: 1.
Further, the step (4) comprises the steps of suspending CTAB-GNRs in physiological saline, dropwise adding PDS-pHPMA-DOX solution under the stirring condition, wherein the molar ratio of CTAB-GNRs to PDS-pHPMA-DOX is 1:1.5, and reacting for a period of time in a dark place to obtain the product pHPMA-DOX @ GNRs.
The third purpose of the invention is realized by adopting the following technical scheme:
application of gold nanorod drug delivery system modified by HPMA polymer in treatment of prostate cancer
Compared with the prior art, the invention has the beneficial effects that: the invention provides a gold nanorod drug loading system modified by a HPMA polymer, wherein a water-soluble HPMA polymer is utilized to assemble chemotherapeutic drugs into GNRs, and the GNRs are cooperated with the effect of thermotherapy chemotherapy. The GNRs absorb near infrared light and convert the near infrared light into heat, the tumor microenvironment is changed by absorbing NIR light, the photothermal conversion effect of the GNRs is efficiently utilized in the process of ensuring the stable release of the chemotherapeutic drugs, the lethality of the chemotherapeutic drugs is improved, the GNRs act on prostate cancer cells in a synergistic manner, and the drug resistance of tumor cells is overcome.
The modification of the pH sensitive HPMA polymer in the drug-loading system improves the stability of GNRs and the tumor accumulation of chemotherapeutic drugs, and reduces the toxicity of the GNRs and the chemotherapeutic drugs to normal tissues. The chemotherapy drug is connected to the HPMA polymer water-soluble bone chain by adopting a pH sensitive hydrazone bond, so that the micromolecule anti-cancer drug can be successfully connected to the HPMA polymer and can be kept stable in blood circulation, the chemotherapy drug can be rapidly released in a tumor cell lysosome and can be transported to a tumor tissue in a targeted way, enters the tumor cell in an endocytosis or pinocytosis mode, can be rapidly released, and the drug effect can be favorably exerted.
The invention also provides a preparation method of the gold nanorod drug delivery system modified by the HPMA polymer, and the preparation process is simple and easy to operate. The invention also provides application of the gold nanorod drug delivery system modified by the HPMA polymer in treating prostate cancer, effectively overcomes drug resistance of tumor cells, and delays occurrence and metastasis of prostate cancer.
Drawings
Fig. 1 is a synthesis scheme of an HPMA polymer-modified gold nanorod drug delivery system of the invention;
FIG. 2 is a transmission electron micrograph of CTAB-GNRs of example 1 and pHPMA-DOX @ GNRs of example 3;
FIG. 3 is a graph of the temperature rise trends of CTAB-GNRs of example 1 and pHPMA-DOX @ GNRs of example 3;
FIG. 4 is a statistical plot of the results of the release of DOX over time of pHPMA-DOX @ GNRs of example 3 in acidic and neutral buffers;
FIG. 5 is a statistical plot of cell viability for 48h incubation of pHPMA-DOX @ GNRs of example 3 with human prostate cancer cells at various concentrations of DOX;
FIG. 6 is a graph of the effect of DOX, pHPMA-DOX @ GNRs of example 3 on the treatment of human prostate cancer cell nude mice.
Detailed Description
The present invention will be further described with reference to the accompanying drawings and the detailed description, and it should be noted that any combination of the embodiments or technical features described below can be used to form a new embodiment without conflict.
Example 1
Preparation of CTAB-GNRs:
(1) preparing a seed solution: under the condition of water bath at 26 ℃, 5mL and 0.2mol/L Cetyl Trimethyl Ammonium Bromide (CTAB) solution are added with 5mL and 0.0005mol/L chloroauric acid solution, after uniform stirring, 600 mu L and 0.01mol/L sodium borohydride solution are rapidly added, the reaction solution is rapidly changed from yellow to brown, the mixture is vigorously stirred for 2min, and the mixture is kept stand for 20-30 min in the dark at 26 ℃.
(2) Preparing a growth solution: under the condition of water bath at 26 ℃, 200 mu L of silver nitrate (AgNO3) solution and 0.004mol/L of silver nitrate solution are sequentially added into 5mL of CTAB solution and 0.2mol/L of CTAB solution, 5mL of chloroauric acid solution and 0.001mol/L of chloroauric acid solution are uniformly stirred, 70 mu L of AA solution and 0.0788mol/L of AA solution are slowly added into the mixture under stirring, the reaction solution is rapidly changed from dark yellow to colorless, 80 mu L of the seed solution is rapidly added into the mixture, the mixture is vigorously stirred for 2min, and the mixture is kept stand overnight at 26 ℃ in a dark place to obtain the CTAB-GNRs.
Example 2
The preparation method of PDS-pHPMA-DOX is shown in figure 1, and comprises the following specific operation steps:
(1) preparation of MA-GG-OH: 8.71ml (0.091mol) of MA-Cl and 30ml of dichloromethane are mixed, 9.79g (0.075mmol) of GG-OH is dissolved in 20ml of 4M sodium hydroxide solution, the dichloromethane solution of the MA-Cl is slowly dropped into the sodium hydroxide solution containing the GG-OH under the stirring at the temperature of minus 15 ℃, simultaneously, 1M sodium hydroxide solution is dropped to adjust the pH value of 9-10, and the mixture is stirred for 1.5h at the room temperature. The dichloromethane layer was separated, washed with 20ml of water, the aqueous layers were combined, and the pH was adjusted to 1-2 with 6M aqueous hydrochloric acid. Recrystallizing the precipitate with 50% ethanol water solution to obtain white crystal MA-GG-OH 6.44g, yield 71.46%, M.p.136-138 deg.C; MS: c8H12N2O4，201.1(M+)。
(2) Preparation of MA-GG-NHNH2: 2.00g (10mmol) of MA-GG-OH are dissolved in 150mL of absolute ethanol, cooled at 0 ℃, added with 2.24g (18mmol) of DIC under stirring, reacted for 2h, and then kept at room temperature for reaction for 2 h. 0.7723g (12mmol) of 80% hydrazine monohydrate was added and the reaction was carried out at room temperature for 4 hours. Adding 150mL of n-hexane, separating out a white precipitate, and washing with an absolute ethanol/n-hexane (v/v is 1:1) mixed solution for 3 times to obtain 1.16g of a white solid, wherein the yield is 54.20%, and the M.p.166-168 ℃; MS: c8H14N4O3，215.0(M+)；
(3) Preparation of MA-GG-NHN ═ DOX: taking MA-GG-NHNH20.15g (0.68mmol) and 0.4g (0.68mmol) of Dox & HCl are dissolved in 35mL of methanol, 1mL of glacial acetic acid is added, stirred at room temperature for 24h, the appropriate amount of polymer containing a hydrazine group is added, and stirred at room temperature for 4h to bind the unreacted Dox & HCl. Separating the product with Sephadex LH-20 Sephadex column, collecting the second red color band, and drying under reduced pressure to obtain red solid product MA-GG-NHN (DOX 142.21 mg), yield of 26%, M.p.108-110 deg.C; MS: c35H41N5O13，740.3(M+)；
(4) Preparation of HPMA: 30ml (0.312mol) of freshly distilled MA-Cl was mixed with 15ml of dichloromethane, slowly added dropwise to 77ml of a dichloromethane solution containing 21ml (0.271mol) of 1-amino-2-propanol and 32g (0.312mol) of sodium carbonate, and after completion of the dropwise addition, the mixture was allowed to warm to room temperature and stirred for 1 hour. Then placing the reaction solution in a low-temperature cooling bath tank at the temperature of 50 ℃ below zero for 1h to obtain the productA white precipitate formed. After filtration, the precipitate was recrystallized from acetone to give 18.04g of white crystals as HPMA, yield 46.31%, mp 66-69 ℃, MS: c7H13NO2，144.1(M+)；
(5) Preparation of PDS-pHPMA-DOX: dissolving 287.27mg (2mmol) of HPMA and 164.60mg (0.22mmol) of MA-GG-NHN (DoX) in 3.82mL of DMSO, adding an initiator ABIK-TT144.73mg, putting into an ampoule bottle, sealing, carrying out oil bath reaction at 50 ℃ for 24h, dissolving the crude product with ultrapure water, dialyzing for 48h, and freeze-drying to obtain a semi-shaking claw polymer TT-pHPMA-DOX180.91mg with a terminal chain of TT; TT-pHPMA-DOX (0.021mM) and PDEA (0.027mM) are taken to be dissolved in 1.75mL of DMF, 0.3mL of DMF containing 5 mu L of DIEA is dripped into the reaction liquid, the mixture is stirred for 3h at room temperature, the solvent is removed under reduced pressure, the crude product is dissolved by deionized water, the dialysis is carried out for 48h, and the product PDS-pHPMA-DOX141.24mg is obtained after freeze drying.
Example 3
The pHPMA-DOX @ GNRs are prepared, the synthetic route is shown in figure 1, and the specific operation steps are as follows:
1mL (0.60nM) of CTAB-GNRs prepared in example 1 was resuspended in physiological saline, and 1mL (0.90nM) of PDS-pHPMA-DOX of example 2 was slowly added dropwise with stirring, and the mixture was left to react for 12h in the dark, to obtain 2mL (0.3nM) of pHPMA-DOX @ GNRs.
Experimental example 1
Transmission electron micrographs for observing CTAB-GNRs and pHPMA-DOX @ GNRs: the transmission electron micrographs of CTAB-GNRs of example 1 and pHPMA-DOX @ GNRs of example 3 were taken, respectively, and it can be seen from FIG. 2A that the CTAB-GNRs have an average transverse length of about 11nm and an average longitudinal length of about 37nm, and from FIG. 2 that the synthesized CTAB-GNRs have a coating of about 5nm formed on the surface of the CTAB-GNRs by the action of the HPMA polymer loaded with doxorubicin.
Experimental example 2
Temperature rising trends for CTAB-GNRs and pHPMA-DOX @ GNRs: respectively sucking 4mL of ultrapure water (Saline), a CTAB-GNRs aqueous solution (0.3nM) and a pHPMA-DOX @ GNRs aqueous solution (0.3nM), placing the ultrapure water, the CTAB-GNRs aqueous solution and the pHPMA-DOX @ GNRs aqueous solution in a quartz cuvette, irradiating the cuvette by using a 808 laser instrument at the temperature of 25 ℃, recording the temperature condition every 1min within 10min, and drawing a temperature rise curve, wherein the result is shown in FIG. 3.
As can be seen from fig. 3: after the gold nanorods are modified by the HPMA-carrying polymer adriamycin conjugates, the medicine carrying system pHPMA-DOX @ GNRs can still convert light energy into heat energy under the NIR condition, the heating effect is similar to that of CTAB-GNRs, and after laser irradiation is carried out for 5min, the temperature can reach more than 40 ℃, so that the pHPMA-DOX @ GNRs have the photo-thermal conversion capability and can play a role in thermal therapy.
Experimental example 3
pHPM A-DOX @ GNRs in vitro drug release: taking a proper amount of the product pHPMA-DOX @ GNRs, dissolving the product pHPMA-DOX @ GNRs in 1mL of acetate buffer solution with pH value of 5.0 or phosphate buffer solution with pH value of 7.4, transferring the product into a dialysis bag (MWCO3500), taking 20mL of corresponding pH buffer solution as a release medium, carrying out release investigation in a shaker (100rpm) at 37 ℃, taking 200 mu L of the release medium at a specific time point, supplementing an equal volume of fresh release medium, centrifuging (1000rpm, 5min), precisely measuring 20 mu L of supernate, carrying out sample injection analysis, and calculating the cumulative release amount at each time point, wherein the result is shown in figure 4.
As can be seen from fig. 4: at pH7.4, the DOX release from pHPMA-DOX @ GNRs was less than 20% within 48h, indicating that the hydrazone bonds remained stable at physiological pH. When the pH is reduced to a weak acid environment in the lysosome, namely pH5.0, the rate of releasing DOX by pHPMA-DOX @ GNRs is obviously accelerated due to the breakage of hydrazone bonds, and the cumulative release amount is 83%. The drug delivery system can keep stable during blood circulation, can ensure that the chemotherapeutic drug is quickly released in a tumor cell lysosome, is transported to a tumor tissue in a targeted manner, enters the tumor cell in an endocytosis or endocytosis mode, can be quickly released, and is beneficial to exerting the drug effect.
Experimental example 4
Cytotoxicity in vitro at various concentrations of pHPMA-DOX @ GNRs: collecting human prostate cancer cell PC-3 (obtained from Shanghai cell bank of Chinese academy of sciences) in DMEM medium (with addition of 10% FBS, 100U/mL penicillin and 100. mu.g/mL streptomycin) at 37 deg.C and 5% CO2Culturing under the conditions of (1).
The PC-3 cell suspension was applied at a density of 1X 104Inoculating the cell/well into a 96-well plate, incubating for 24h, discarding the culture medium, adding culture medium containing DOX or pHPMA-DOX @ GNRs with different concentrations, setting five multiple wells for each group of concentration, and incubating the cells with fresh culture mediumAs a negative control, the cell-free medium was used as a blank, incubated for 48h, 20. mu.L of MTT in PBS (5mg/mL) was added to each well, incubated for 4h, the solution was aspirated, 150. mu.L of DMSO was added, shaken for 20min, and the absorbance at 570nm of each well was measured using a microplate reader to calculate the cell viability, where pHPMA-DOX @ GNRs + Laser indicates that the Laser irradiation was 808 min after 2h of pHPMA-DOX @ GNRs administration, and the results are shown in FIG. 5.
As can be seen from fig. 5: the cytotoxicity of the pHPMA-DOX @ GNRs is far less than that of DOX, but the growth inhibition activity of the pHPMA-DOX @ GNRs on PC-3 cells is positively correlated with the concentration after NIR illumination. Under high concentration conditions, pHPMA-DOX @ GNRs + Laser has a significantly higher effect on the survival rate of PC-3 than DOX. The drug delivery system improves the stability of GNRs and the tumor accumulation of doxorubicin hydrochloride by modifying the pH sensitive HPMA polymer, and reduces the toxicity of the GNRs and the doxorubicin hydrochloride to normal tissues.
Experimental example 5
In vivo anti-tumor effects of pHPMA-DOX @ GNRs: 100 μ L of PC-3 cell suspension (6X 106cells, PBS) was injected subcutaneously into one shoulder of nude mice of male mice to establish a PC-3-bearing unilateral tumor model. When the tumor volume reaches 100-200mm3At this time, the tumor sites were irradiated with NIR at 808nm, 2W/cm after administration of pHPMA-DOX @ GNRs + Laser (pHPMA-DOX @ GNRs after 2 h), which were randomly divided into 5 groups (n ═ 6) each of physiological Saline (Saline), DOX, pHPMA-DOX @ GNRs, and combination treatment groups pHPMA-DOX @ GNRs + Laser (pHPMA-DOX @ GNRs were administered)25 min). Tail vein injections were given corresponding drug treatments at days 1, 8 and 15 with DOX equivalent of 5 mg/kg. Body weight and tumor size (i.e., tumor length and width) of nude mice were measured every three days for plotting the change in body weight and tumor volume, and the results are shown in fig. 6.
As can be seen from fig. 6: tumor growth was inhibited in the DOX group, pHPMA-DOX @ GNRs group and the combination-treated pHPMA-DOX @ GNRs + Laser group relative to the control group (Saline group). Statistically, the tumor volumes of the pHPMA-DOX @ GNRs group and the combination treatment group were significantly smaller than that of the control group (p <0.01), and the tumors of some mice in the combination treatment group were completely cured. The DOX tumor growth inhibition rate (TGI) is 31 percent, and the pharmaceutical composition has moderate anti-tumor effect, while the TGI of pHPMA-DOX @ GNRs is improved by one time and is about 68 percent, the good anti-tumor effect is attributed to longer blood circulation and stronger EPR effect, and the TGI of a combined treatment group reaches 88 percent, which shows that the drug carrying system of the invention utilizes the water-soluble HPMA polymer to assemble chemotherapeutic drugs into GNRs to cooperate with thermotherapy and chemotherapy. The GNRs absorb near infrared light and convert the near infrared light into heat, the tumor microenvironment is changed by absorbing NIR light, the photothermal conversion effect of the GNRs is efficiently utilized in the process of ensuring the stable release of the chemotherapeutic drugs, the lethality of the chemotherapeutic drugs is improved, the drug resistance of tumor cells is effectively overcome by the synergistic effect and the tumor cells of the prostate cancer, and the potential advantages of the drug delivery system in the chemical-photothermal therapy of the prostate cancer are further shown.
The above embodiments are only preferred embodiments of the present invention, and the protection scope of the present invention is not limited thereby, and any insubstantial changes and substitutions made by those skilled in the art based on the present invention are within the protection scope of the present invention.
Claims (7)
1. The gold nanorod drug loading system modified by the HPMA polymer is characterized in that the drug loading system takes the pH sensitive HPMA polymer as a modification chain of the gold nanorod, and the pH sensitive HPMA polymer is connected with a chemotherapeutic drug through a pH sensitive chemical bond;
the pH sensitive chemical bond is a hydrazone bond;
the chemotherapeutic drug is adriamycin;
the preparation method of the gold nanorod drug delivery system modified by the HPMA polymer comprises the following steps:
(1) preparing CTAB-GNRs by adopting a seed solution growth method;
(2) taking MA-GG-NHNH2Reacting with chemotherapeutic drug adriamycin to obtain intermediate MA-GG-NHN ═ DOX;
(3) preparing a pH-sensitive HPMA polymer PDS-pHPMA-DOX by using HPMA and the intermediate MA-GG-NHN ═ DOX obtained in the step (2) as monomers;
(4) and (3) mixing the CTAB-GNRs prepared in the step (1) with the product PDS-pHPMA-DOX prepared in the step (3), and reacting in a dark place to obtain the gold nanorod drug delivery system pHPMA-DOX @ GNRs modified by the HPMA polymer.
2. The HPMA polymer-modified gold nanorod drug delivery system of claim 1, wherein the MA-GG-NHN ═ DOX preparation process in step (2) is as follows: taking MA-GG-NHNH2Dissolving adriamycin in methanol, adding glacial acetic acid, and stirring at room temperature to obtain the product.
3. The HPMA polymer-modified gold nanorod drug delivery system of claim 2, wherein the MA-GG-NHNH is configured to carry out drug delivery in a single cell2And doxorubicin at a 1:1 molar ratio.
4. The HPMA polymer modified gold nanorod drug delivery system of claim 1, wherein step (3) comprises the following steps: 4,4 '-azobis (cyanovaleric acid) -thiazolidine-2-thione is used as an initiator, DMSO is used as a reaction solvent, 4' -azobis (cyanovaleric acid) -thiazolidine-2-thione, DMSO, monomers HPMA, MA-GG-NHN ═ DOX are placed in an ampoule bottle and sealed, oil bath reaction is carried out at 50 ℃ for 24 hours, ultrapure water is used for dissolving a crude product, dialysis is carried out for 48 hours, and freeze drying is carried out to obtain a half-shaking claw polymer TT-pHPMA-DOX with a chain at the tail end being TT; TT-pHPMA-DOX and PDEA are dissolved in DMF, DMF containing DIEA is dripped into reaction liquid, the mixture is stirred for 3 hours at room temperature, the solvent is removed under reduced pressure, deionized water is used for dissolving the crude product, dialysis is carried out for 48 hours, and the polymer PDS-pHPMA-DOX is obtained after freeze drying.
5. The HPMA polymer modified gold nanorod drug delivery system of claim 4, wherein the weight percentage of the mixture of AIBK-TT, DMSO, monomeric HPMA, and MA-GG-NHN ═ DOX is 2.4%, 85.1%, 12.5%, respectively, wherein the molar ratio of monomeric HPMA and MA-GG-NHN ═ DOX is 9: 1.
6. The HPMA polymer modified gold nanorod drug delivery system according to claim 1, wherein the step (4) comprises resuspending CTAB-GNRs in physiological saline, adding a PDS-pHPMA-DOX solution dropwise under stirring, wherein the molar ratio of the CTAB-GNRs to the PDS-pHPMA-DOX is 1:1.5, and reacting for a period of time in a dark place to obtain the product pHPMA-DOX @ GNRs.
7. Use of the HPMA polymer-modified gold nanorod drug delivery system of claim 1 in the preparation of a medicament for the treatment of prostate cancer.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2012009406A3 (en) * | 2010-07-13 | 2012-04-12 | University Of Utah Research Foundation | Gold particles and methods of making and using the same in cancer treatment |
| CN102858323A (en) * | 2010-04-15 | 2013-01-02 | 浦项工科大学校产学协力团 | Anticancer drug delivery system using ph-sensitive metal nanoparticles |
| CN104758948A (en) * | 2015-04-25 | 2015-07-08 | 郑州大学 | Preparation method and application of gold nano-star based multi-functional anti-tumor target diagnostic treatment medicine |
-
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102858323A (en) * | 2010-04-15 | 2013-01-02 | 浦项工科大学校产学协力团 | Anticancer drug delivery system using ph-sensitive metal nanoparticles |
| WO2012009406A3 (en) * | 2010-07-13 | 2012-04-12 | University Of Utah Research Foundation | Gold particles and methods of making and using the same in cancer treatment |
| CN104758948A (en) * | 2015-04-25 | 2015-07-08 | 郑州大学 | Preparation method and application of gold nano-star based multi-functional anti-tumor target diagnostic treatment medicine |
Non-Patent Citations (2)
| Title |
|---|
| Enhanced Reactive Oxygen Species Generation by Mitochondria Targeting of Anticancer Drug To Overcome Tumor Multidrug Resistance;Liu Y等;《Biomacromolecules》;20190829;摘要,表1，图示2 * |
| PEGylated PAMAM dendrimer-doxorubicin conjugate-hybridized gold nanorod for combined photothermal-chemotherapy;Li X等;《Biomaterials》;20140509;摘要 * |
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