CN114748644B - Preparation method of phycocyanin molecularly imprinted drug carrier with ZIF-8 as matrix - Google Patents
Preparation method of phycocyanin molecularly imprinted drug carrier with ZIF-8 as matrix Download PDFInfo
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Abstract
The invention discloses a preparation method of an phycocyanin molecularly imprinted drug carrier with ZIF-8 as a matrix, belonging to the field of functional material preparation. The invention aims to solve the problems of poor drug synergistic effect, low drug loading rate, easy early leakage of drugs, limited targeting and large toxic and side effects of the existing drug carrier. The method comprises the following steps: the invention encapsulates doxorubicin in ZIF-8 as a carrier matrix, adopts surface imprinting polymerization, takes phycocyanin as a template molecule, and does not elute the template molecule to prepare the drug-loaded imprinted polymer. On the basis, modifying PEG sequence connected with folic acid, doping calcium carbonate, and preparing the targeting drug-carrying molecularly imprinted polymer. The drug carrier disclosed by the invention has the advantages of excellent drug loading rate, good drug release effect and biodegradability, and can be used for dual drug synergistic treatment of targeted cancer cells. Can be used as a drug delivery system of compound preparation.
Description
Technical Field
The invention belongs to the field of functional material preparation, and particularly relates to a preparation method of an phycocyanin molecularly imprinted drug carrier with ZIF-8 as a matrix.
Background
Doxorubicin is a broad-spectrum antitumor drug which has a certain curative effect on cancers such as breast cancer, sarcoma, lung cancer, bladder cancer and the like, but has a strong cytotoxicity effect, so that doxorubicin is often used in combination with other anticancer drugs. Phycocyanin is the most effective and representative of the anti-tumor drugs of algae, has good biological activity, can improve the activity of regulating enzymes and the synthesis of various proteins, has good anti-tumor activity, has obvious killing effect on various cancer cell lines, has small toxic and side effects, has no influence on normal cell tissues, and has a protective effect on apoptosis and death caused by free radicals. The combination of phycocyanin and doxorubicin can significantly enhance therapeutic effect by synergistic effect of anticancer drug, and minimize adverse side effects of drug without being affected by dosage.
However, there are some limitations of general drug delivery vehicles, mainly related to insufficient drug loading, difficulty in responding to drug release and susceptibility to drug leakage. Molecularly Imprinted Polymers (MIPs) can selectively recognize and carry template molecules and structural analogs thereof. Due to their unique properties, such as selective recognition capability and controlled drug release initiated by imprinted sites in the polymer structure, are often selected as drug delivery vehicles. However, the imprinting of two template molecules is easy to cause the problems of uneven imprinting site distribution, mutual occupation of imprinting sites and the like when two drugs are loaded simultaneously. The zeolite imidazole ester skeleton series material (ZIFs) is a porous crystalline material with a zeolite-like structure, can belong to a special metal organic framework MOFs material, has a tetrahedral three-dimensional network structure, and has the advantages of larger specific surface area, high porosity, ultrahigh drug loading efficiency, biodegradability, good biocompatibility and the like; however, its stability and ease of drug leakage limit its practical application as a drug carrier.
Disclosure of Invention
The invention provides a preparation method of an phycocyanin molecularly imprinted drug carrier taking ZIF-8 as a matrix, which aims to solve the problems of poor drug synergistic effect, low drug loading rate, easy early leakage of drugs, limited targeting and large toxic and side effects of the existing drug carrier.
The preparation method of the phycocyanin molecularly imprinted drug carrier with ZIF-8 as a matrix comprises the following steps:
1. preparation of ZIF-8:
zn (NO) 3 ) 2 ·6H 2 Respectively adding O and 2-methylimidazole into deionized water, and performing ultrasonic treatment for 3-6 min to obtain Zn (NO) 3 ) 2 ·6H 2 O solution and 2-methylimidazole solution; zn (NO) was taken up at room temperature 3 ) 2 ·6H 2 Mixing and stirring the O solution and the 2-methylimidazole solution, washing the mixed solution by water and ethanol in sequence, and vacuum drying to obtain ZIF-8; the Zn (NO) 3 ) 2 ·6H 2 The mass ratio of O to 2-methylimidazole is 1 (8-12); the Zn (NO) 3 ) 2 ·6H 2 The concentration of the O solution is 0.01-0.015 g/mL; the concentration of the 2-methylimidazole solution is 0.1-0.15 g/mL;
2. preparation of ZIF-8/DOX:
dispersing DOX and ZIF-8 in PBS solution, and carrying out ultrasonic treatment for 50-70 min; stirring for 22-26 hours in a dark place, centrifuging, and collecting supernatant to obtain ZIF-8/DOX; the volume ratio of the DOX to the PBS solution is 1g (1200-1400 mL); the volume ratio of the ZIF-8 to the PBS solution is 1g (180-220) mL;
3. preparation of ZIF-8/DOX@MIPs-FA:
dispersing ZIF-8/DOX in PBS solution, and then sequentially adding phycocyanin and alpha-methacrylic acid to obtain a mixed solution; magnetically stirring the mixed solution for 0.8-1.2 h, and introducing N 2 Removing oxygen, sequentially adding N-isopropyl acrylamide, polyethylene glycol 2000 and ammonium persulfate, performing polymerization reaction at 20-30 ℃ for 22-26 hours, and then adding PEG-FA and CaCO 3 Continuously reacting for 4-6 hours under magnetic stirring, washing with deionized water, and vacuum drying to obtain ZIF-8/DOX@MIPs-FA; the volume ratio of the ZIF-8/DOX to the PBS solution is 1g (180-220) mL; the volume ratio of the phycocyanin to the PBS solution is 1g (900-1100 mL); the volume ratio of the alpha-methacrylic acid to the PBS solution is 1g (350-400 mL); the volume ratio of the N-isopropyl acrylamide to the PBS solution is 1g (280-320 mL); the volume ratio of the polyethylene glycol 2000 to the PBS solution is 1g (180-220) mL; the volume ratio of the ammonium persulfate to the PBS solution is 1g (1700-1900) mL; body of the PEG-FA and PBS solutionThe product ratio is 1g (90-110) mL; said CaCO 3 The volume ratio of the solution to the PBS solution is 1g (3800-4200) mL.
The invention has the advantages that:
1. the drug carrier prepared by the invention can simultaneously have higher loading capacity for two anticancer drugs, and reduce the toxic and side effects of doxorubicin on normal cells when being used for synergizing anticancer.
2. The invention applies the molecular imprinting technology to prepare the drug-loaded imprinted polymer on the surface of the drug-loaded ZIF-8. The surface imprinting polymer layer has the effects of stabilizing ZIF-8 and preventing the drug from leaking in advance so as to respond to drug release in the tumor microenvironment.
3. According to the invention, doxorubicin is loaded in ZIF-8, an phycocyanin molecularly imprinted polymer is prepared on the surface of the doxorubicin, and PEG-FA is modified in a non-imprinted region, so that good biocompatibility and targeting property are obtained. Avoiding mutual interference caused by imprinting site effect of targeting recognition and drug release, and avoiding solvent use and residue reduction of secondary drug imprinting.
4. The doping of calcium carbonate plays a role in protecting the surface medicine of the imprinted polymer, and after the phycocyanin in the imprinted sites are released in response to the tumor microenvironment, the imprinted sites are exposed to promote the effective release of the internal doxorubicin, so that the synergistic release of the doxorubicin and the phycocyanin is ensured, and the toxic and side effects on normal cells are reduced.
5. The invention also solves the problem that the existing drug carrier is difficult to biodegrade.
Drawings
FIG. 1 is a bar graph comparing the drug loading and encapsulation rates of ZIF-8 (DOX) @ MIPs-FA for DOX and PC;
FIG. 2 is a graph showing comparison of DOX and PC release levels in ZIF-8 (DOX) @ MIPs-FA at pH values of 5.0 (a) and 7.4 (b);
FIG. 3 is a histogram comparing cytotoxicity of ZIF-8, ZIF-8 (DOX) and ZIF-8 (DOX) @ MIPs-FA against RWPE-2 of normal prostate cells;
FIG. 4 is a histogram comparing cytotoxicity of ZIF-8, ZIF-8 (DOX) and ZIF-8 (DOX) @ MIPs-FA against prostate cancer cells DU 145;
FIG. 5 is a graph of the residual mass percent in ZIF-8 (DOX) @ MIPs-FA degradation in vitro.
Detailed Description
The first embodiment is as follows: the preparation method of the phycocyanin molecularly imprinted drug carrier with ZIF-8 as a matrix in the embodiment specifically comprises the following steps:
1. preparation of ZIF-8:
zn (NO) 3 ) 2 ·6H 2 Respectively adding O and 2-methylimidazole into deionized water, and performing ultrasonic treatment for 3-6 min to obtain Zn (NO) 3 ) 2 ·6H 2 O solution and 2-methylimidazole solution; zn (NO) was taken up at room temperature 3 ) 2 ·6H 2 Mixing and stirring the O solution and the 2-methylimidazole solution, washing the mixed solution by water and ethanol in sequence, and vacuum drying to obtain ZIF-8; the Zn (NO) 3 ) 2 ·6H 2 The mass ratio of O to 2-methylimidazole is 1 (8-12); the Zn (NO) 3 ) 2 ·6H 2 The concentration of the O solution is 0.01-0.015 g/mL; the concentration of the 2-methylimidazole solution is 0.1-0.15 g/mL;
2. preparation of ZIF-8/DOX:
dispersing DOX and ZIF-8 in PBS solution, and carrying out ultrasonic treatment for 50-70 min; stirring for 22-26 hours in a dark place, centrifuging, and collecting supernatant to obtain ZIF-8/DOX; determining the drug loading rate and the drug encapsulation rate of the nano particles by adopting an ultraviolet spectrophotometry; the volume ratio of the DOX to the PBS solution is 1g (1200-1400 mL); the volume ratio of the ZIF-8 to the PBS solution is 1g (180-220) mL;
3. preparation of ZIF-8/DOX@MIPs-FA:
dispersing ZIF-8/DOX in PBS solution, and then sequentially adding phycocyanin and alpha-methacrylic acid to obtain a mixed solution; magnetically stirring the mixed solution for 0.8-1.2 h, and introducing N 2 Removing oxygen, sequentially adding N-isopropyl acrylamide, polyethylene glycol 2000 and ammonium persulfate, performing polymerization reaction at 20-30 ℃ for 22-26 hours, and then adding PEG-FA and CaCO 3 Continuously reacting for 4-6 hours under magnetic stirring, washing with deionized water, and vacuum drying to obtain ZIF-8/DOX@MIPs-FA; the volume ratio of the ZIF-8/DOX to the PBS solution is 1g (180-220) mL;the volume ratio of the phycocyanin to the PBS solution is 1g (900-1100 mL); the volume ratio of the alpha-methacrylic acid to the PBS solution is 1g (350-400 mL); the volume ratio of the N-isopropyl acrylamide to the PBS solution is 1g (280-320 mL); the volume ratio of the polyethylene glycol 2000 to the PBS solution is 1g (180-220) mL; the volume ratio of the ammonium persulfate to the PBS solution is 1g (1700-1900) mL; the volume ratio of the PEG-FA to the PBS solution is 1g (90-110) mL; said CaCO 3 The volume ratio of the solution to the PBS solution is 1g (3800-4200) mL.
In this embodiment, DOX is doxorubicin.
The embodiment creatively provides a method for loading the antitumor drug doxorubicin by taking the degradable metal organic framework ZIF-8 as a drug-carrying matrix, and on the basis, the template molecule phycocyanin is not eluted, so that the drug-carrying MIPs are further synthesized, the drug carrying capacity is increased, the drug is protected, the drug is prevented from leaking in advance, and the efficiency of drug delivery and cancer treatment is improved. DOX is encapsulated in the carrier matrix of MIPs, and the polymer carrier matrix can slowly diffuse and release the encapsulated anticancer drug through the decomposition of the polymer coating. ZIF-8 is added in the polymerization process, so that the specific surface area is increased, the accessibility of imprinting sites is improved, the embedding of template molecules is reduced, and the preparation method has the degradable characteristics and the controllable drug release capability. The non-imprinted region can be further targeted and modified to enable the surface of the material to be functionalized to bind to the targeting ligand. In this way, the polymer may allow the ligand to recognize a receptor overexpressed on the surface of a particular cancer cell for active targeting by receptor-mediated endocytosis. Most tumors in humans, such as ovarian, breast, lung, and brain cancers, are highly expressed by the cell surface Folate Receptor (FRs), and moreover, the targeting ligand Folic Acid (FA) is non-immunogenic, stable during storage, low in molecular weight, and readily diffusible through biological barriers. Polyethylene glycol (PEG) is a high molecular polymer with good water solubility, biocompatibility and dispersibility. In biomedicine, PEG is commonly used for modifying a drug carrier, so that the stability of the drug carrier in blood is enhanced, the blood circulation of the drug carrier is prolonged, and the drug concentration is increased in an accumulated way. PEG and FA are connected in sequence, and folic acid side chain can enter target cells through folic acid receptor to transfer medicine to tumor site.
In the embodiment, DOX is encapsulated in ZIF-8 to be used as a bearing matrix, and a surface imprinting polymerization method is adopted, phycocyanin (PC) is used as a template molecule, and methacrylic acid is used as a functional monomer to prepare the molecularly imprinted polymer. Based on the method, FA is connected through a PEG sequence, calcium carbonate is doped, and the response drug release targeting drug-loaded molecularly imprinted polymer (ZIF-8 (DOX) @ MIPs-FA) is prepared. PC acts as an anti-tumor drug, enhances the anti-tumor effect of DOX, and reduces the side effects of DOX. The dual-drug simultaneous carrying and FA targeting functions enable two anticancer drugs to reach the disease position accurately and simultaneously, respond to drug release, and better exert synergistic effect. PEG improves the hydrophilicity of ZIF-8 (DOX) @ MIPs-FA, enhances its stability in biological systems, and prolongs its circulation time, thereby increasing the drug concentration around cancer cells. ZIF-8 (DOX) @ MIPs-FA gives consideration to superior drug loading, higher biocompatibility, good drug release effect and accurate targeting functionality. The research provides a new strategy for effective drug synergistic treatment, and provides an effective new idea for developing a drug delivery system of a compound formula.
The second embodiment is as follows: the first difference between this embodiment and the specific embodiment is that: the mixing and stirring in the first step is vigorously stirring at 1000rpm for 30min. The other is the same as in the first embodiment.
And a third specific embodiment: this embodiment differs from one of the first to second embodiments in that: the vacuum drying in the first step is vacuum drying at 50 ℃ for 12 hours. The other is the same as in one of the first to second embodiments.
The specific embodiment IV is as follows: this embodiment differs from one of the first to third embodiments in that: in step one, zn (NO 3 ) 2 ·6H 2 The mass ratio of O to 2-methylimidazole is 1 (10); the Zn (NO) 3 ) 2 ·6H 2 The concentration of the O solution is 0.0125g/mL; the concentration of the 2-methylimidazole solution was 0.125g/mL. The other is the same as in one of the first to third embodiments.
Fifth embodiment: this embodiment differs from one to four embodiments in that: the PBS solution in the second step has pH of 7 and concentration of 20mmol.L -1 . The others are the same as in one to one fourth embodiments.
Specific embodiment six: this embodiment differs from one of the first to fifth embodiments in that: the centrifugation in the second step is at 5000rpm for 20min. The others are the same as in one of the first to fifth embodiments.
Seventh embodiment: this embodiment differs from one of the first to sixth embodiments in that: in the second step, the volume ratio of the mass of DOX to the PBS solution is 0.75 g/1000 mL; the volume ratio of the ZIF-8 to the PBS solution is 1 g/200 mL. The others are the same as in one of the first to sixth embodiments.
Eighth embodiment: this embodiment differs from one of the first to sixth embodiments in that: in the third step, the volume ratio of the ZIF-8/DOX to the PBS solution is 1g:200mL; the volume ratio of the phycocyanin to the PBS solution is 1g to 1000mL; the volume ratio of the alpha-methacrylic acid to the PBS solution is 26g:10000mL. The others are the same as in one of the first to sixth embodiments.
Detailed description nine: this embodiment differs from one of the first to sixth embodiments in that: the volume ratio of the N-isopropyl acrylamide to the PBS solution in the third step is 34g:10000mL; the volume ratio of the polyethylene glycol 2000 to the PBS solution is 1g:200mL; the volume ratio of ammonium persulfate to PBS solution was 5.5g:10000mL. The others are the same as in one of the first to sixth embodiments.
Detailed description ten: this embodiment differs from one of the first to sixth embodiments in that: mw=2000 Da of the PEG-FA in step three; the volume ratio of the PEG-FA to the PBS solution is 1g to 100mL; said CaCO 3 The volume ratio to PBS solution was 1g:4000mL. The others are the same as in one of the first to sixth embodiments.
The beneficial effects of the invention are verified by the following examples:
example 1: the preparation method of the phycocyanin molecularly imprinted drug carrier with ZIF-8 as a matrix comprises the following steps:
1. preparation of ZIF-8:
1.0g Zn (NO) 3 ) 2 ·6H 2 Adding O and 10g of 2-methylimidazole into 80mL of deionized water respectively, and performing ultrasonic treatment for 5min to obtain Zn (NO) 3 ) 2 ·6H 2 O solution and 2-methylimidazole solution; zn (NO) was taken up at room temperature 3 ) 2 ·6H 2 Mixing the O solution and the 2-methylimidazole solution, vigorously stirring at 1000rpm for 30min, washing the mixed solution with water and ethanol in sequence, and then vacuum drying at 50 ℃ for 12h to obtain ZIF-8;
2. preparation of ZIF-8 (DOX):
15mg DOX and 100mg ZIF-8 were dispersed in 20mL PBS solution (pH=7.0, 20 mmol.L) -1 ) Ultrasonic treatment is carried out for 60 min; stirring for 24h in dark, centrifuging at 5000rpm for 20min, and collecting supernatant after centrifuging to obtain ZIF-8 (DOX);
3. preparation of ZIF-8 (DOX) @ MIPs-FA:
100mg ZIF-8 (DOX) was dispersed in 20mL PBS solution (pH=7.0, 20 mmol.L) -1 ) Then 20mg phycocyanin and 52mg (6 mmol) of alpha-methacrylic acid are added in sequence to obtain a mixed solution; magnetically stirring the mixed solution for 1h, and introducing N 2 After removing oxygen, 68mg (6 mmol) of N-isopropylacrylamide, 100mg of polyethylene glycol 2000 and 11mg of ammonium persulfate were sequentially added thereto for polymerization at 25℃for 24 hours, and 200mg of PEG-FA (MW=2000 Da) and 5mg of CaCO were further added thereto 3 And continuing to react for 5 hours under magnetic stirring, washing with deionized water to remove PBS and other residual components, and vacuum drying to obtain ZIF-8 (DOX) @ MIPs-FA.
1. Study of the drug-loading amount of ZIF-8- (DOX) @ MIPs-FA
Drug Loading (DLC) and Drug Encapsulation Efficiency (DEE) formulas (2-1), (2-2) are as follows:
(2-1)
(2-2)
wherein M is 1 (mg) is the mass of the supernatant Chinese medicine, M 0 (mg) is the mass of the drug added, M is the sum of the mass of the encapsulated drug and the mass of the loading material added.
As shown in FIG. 1, the ZIF-8 (DOX) @ MIPs-FA has higher drug loading rate and drug encapsulation rate to DOX and PC, because the ZIF-8 (DOX) @ MIPs-FA uses ZIF-8 as a bearing matrix, the specific surface area is increased, and the drug loading efficiency is improved. Doxorubicin is loaded in ZIF-8, phycocyanin molecularly imprinted polymer is prepared on the surface of the doxorubicin, the two medicaments are not interfered with each other in the loading process, the preparation is simple, and the use of solvents and elution of template molecules are reduced. DOX is well coated in the polymer layer, and meanwhile, calcium carbonate has a certain protection effect on the drug PC before the PC is released, and the preparation scheme avoids the premature leakage of the drug.
2. In vitro drug release experiment
ZIF-8 (DOX) @ MIPs loaded with DOX and PC were dispersed in PBS solution at pH 5.0 or 7.4. The above system was centrifuged at 37.+ -. 1 ℃ for 10min at specified time intervals and the released drug concentration was determined. And the mass remaining of the material was measured.
To mimic the drug release behavior of ZIF-8 (DOX) @ MIPs-FA in tumor stroma or tumor cells and intravenous injection. In this study, in vitro drug release of drug-loaded ZIF-8 (DOX) @ MIPs-FA was examined in two buffer solutions with pH values of 5.0 and 7.4, respectively, and FIG. 2 shows the amounts of released ZIF-8 (DOX) @ MIPs-FA of DOX and PC in different drug release conditions. The release amounts of DOX and PC in ZIF-8 (DOX) @ MIPs were 100mg g, respectively, at a pH of 5.0 in the buffer solution -1 And 135 mg g -1 Are higher than the drug release amount under the condition that the pH value of the buffer solution is 7.4. According to the results, there was a significant difference in the cumulative drug release at different pH values. The total drug release in the buffer solution at pH 5.0 is much higher than in the buffer solution at pH 7.0, which is caused by the dissolution of calcium carbonate on the surface of ZIF-8 (DOX) @ MIPs-FA due to the slightly acidic environmentThe carboxyl group of MAA protonates to disrupt the interaction between the functional monomer and the template molecule and thus effect drug release. The drugs in the PC sites on the surfaces of ZIF-8 (DOX) @ MIPs-FA are exposed to the outside and released first, so that the toxic and side effects caused by the early release of doxorubicin are avoided. The exposed blotting site after PC release causes ZIF-8 to be exposed to a slightly acidic environment, thereby further effectively releasing the drug doxorubicin. It is well known that the pH value of the microenvironment around cancer cells is about 5.0, so ZIF-8 (DOX) @ MIPs-FA prepared by the research can be decomposed with calcium carbonate in the slightly acidic condition of the cancer cells, and metal chelation and hydrogen bond destruction between functional monomers and drugs are carried out, thereby achieving the controlled release of the drugs, avoiding the release in normal tissues and reducing the killing property on the normal cells. And the prepared ZIF-8 (DOX) @ MIPs-FA can be subjected to framework decomposition in a tumor slightly acidic environment, so that the deep-coated medicine DOX is released more completely, and the mutual synergistic effect of medicines is exerted.
3. Cytotoxicity study
To investigate the cytotoxicity of ZIF-8 (DOX) @ MIPs-FA on tumor cells, MTT assays were performed. Prostate cancer cells DU 145 and normal prostate cells RWPE-2 were used at 1X 10 per well 4 The density of individual cells was inoculated in medium (80. Mu.L) and incubated at 37℃for 20 h. Next, the medium was replaced with 80. Mu.L of a different fresh medium containing the specific reagent (ZIF-8, ZIF-8 (DOX) @ MIPs-FA) and incubated 5 h. Subsequently, the medium was changed to contain 3- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide (MTT) (final concentration of 0.5 mg. Multidot.mL) -1 ) Is used as a fresh medium for the culture. Cells were re-incubated 3 h. Then, the supernatant was replaced with DMSO (80. Mu.L) and shaken on a shaker for 10 min. The absorbance of the solution was obtained by a microplate reader to measure the Optical Density (OD) value. Cell viability was calculated according to formula (2-3):
(2-3)
wherein OD treated Obtained from cells treated with a specific drug, OD control Is never processed finelyObtained in cells.
The relative cell viability of ZIF-8, ZIF-8 (DOX) and ZIF-8 (DOX) @ MIPs-FA treated prostate cancer cells DU 145 and normal prostate cells RWPE-2, respectively, is shown in FIGS. 3 and 4. After 5 hours of incubation, the DOX-loaded drug carrier ZIF-8 (DOX) had some killing power on cancer cells, indicating that the killing effect on cells is caused by the anticancer drug DOX. Compared with ZIF-8 (DOX) @ MIPs and ZIF-8 (DOX), ZIF-8 (DOX) @ MIPs-FA has larger cancer cytotoxicity to cancer cells and lower toxic and side effects to normal cells at various concentrations, because the surface of ZIF-8 (DOX) @ MIPs-FA is imprinted with PC drugs, and the ZIF-8 (DOX) @ MIPs-FA has targeted enrichment effect around cancer cells, thereby enhancing the killing effect to cancer cells and reducing the toxic and side effects of drugs to normal cells.
4. Degradation study
As can be seen from FIG. 5, ZIF-8- (DOX) @ MIPs-FA was degraded in PBS solution at pH 5.0 for about 77% of the remaining mass, the degradation started with calcium carbonate aspiration and drug release, and then the metal chelation of the carboxyl group of methacrylic acid in the polymer with the metal Zn ion in ZIF-8 was broken down, the imprinting layer was decomposed, ZIF-8 was gradually biodegraded, and ZIF-8- (DOX) @ MIPs-FA showed good biodegradability.
Claims (10)
1. The preparation method of the phycocyanin molecular imprinting drug carrier taking ZIF-8 as a matrix is characterized by comprising the following steps of:
1. preparation of ZIF-8:
zn (NO) 3 ) 2 ·6H 2 Respectively adding O and 2-methylimidazole into deionized water, and performing ultrasonic treatment for 3-6 min to obtain Zn (NO) 3 ) 2 ·6H 2 O solution and 2-methylimidazole solution; zn (NO) was taken up at room temperature 3 ) 2 ·6H 2 Mixing O solution and 2-methylimidazole solution, stirring, and mixingWashing the liquid sequentially with water and ethanol, and vacuum drying to obtain ZIF-8; the Zn (NO) 3 ) 2 ·6H 2 The mass ratio of O to 2-methylimidazole is 1 (8-12); the Zn (NO) 3 ) 2 ·6H 2 The concentration of the O solution is 0.01-0.015 g/mL; the concentration of the 2-methylimidazole solution is 0.1-0.15 g/mL;
2. preparation of ZIF-8/DOX:
dispersing DOX and ZIF-8 in PBS solution, and carrying out ultrasonic treatment for 50-70 min; stirring for 22-26 hours in a dark place, centrifuging, and collecting supernatant to obtain ZIF-8/DOX; the volume ratio of the DOX to the PBS solution is 1g (1200-1400 mL); the volume ratio of the ZIF-8 to the PBS solution is 1g (180-220) mL;
3. preparation of ZIF-8/DOX@MIPs-FA:
dispersing ZIF-8/DOX in PBS solution, and then sequentially adding phycocyanin and alpha-methacrylic acid to obtain a mixed solution; magnetically stirring the mixed solution for 0.8-1.2 h, and introducing N 2 Removing oxygen, sequentially adding N-isopropyl acrylamide, polyethylene glycol 2000 and ammonium persulfate, performing polymerization reaction at 20-30 ℃ for 22-26 hours, and then adding PEG-FA and CaCO 3 Continuously reacting for 4-6 hours under magnetic stirring, washing with deionized water, and vacuum drying to obtain ZIF-8/DOX@MIPs-FA; the volume ratio of the ZIF-8/DOX to the PBS solution is 1g (180-220) mL; the volume ratio of the phycocyanin to the PBS solution is 1g (900-1100 mL); the volume ratio of the alpha-methacrylic acid to the PBS solution is 1g (350-400 mL); the volume ratio of the N-isopropyl acrylamide to the PBS solution is 1g (280-320 mL); the volume ratio of the polyethylene glycol 2000 to the PBS solution is 1g (180-220) mL; the volume ratio of the ammonium persulfate to the PBS solution is 1g (1700-1900) mL; the volume ratio of the PEG-FA to the PBS solution is 1g (90-110) mL; said CaCO 3 The volume ratio of the solution to the PBS solution is 1g (3800-4200) mL.
2. The method for preparing a ZIF-8-based phycocyanin molecularly imprinted drug carrier according to claim 1, wherein the mixing and stirring in the first step is vigorously stirring at 1000rpm for 30min.
3. The method for preparing a ZIF-8-based phycocyanin molecularly imprinted drug carrier according to claim 1, wherein the vacuum drying in the first step is vacuum drying at 50 ℃ for 12h.
4. The method for preparing a ZIF-8-based phycocyanin molecularly imprinted drug carrier according to claim 1, wherein the Zn (NO 3 ) 2 ·6H 2 The mass ratio of O to 2-methylimidazole is 1 (10); the Zn (NO) 3 ) 2 ·6H 2 The concentration of the O solution is 0.0125g/mL; the concentration of the 2-methylimidazole solution was 0.125g/mL.
5. The preparation process of phycocyanin molecular engram medicine carrier with ZIF-8 as matrix as claimed in claim 1, wherein the PBS solution in the second step has pH of 7 and concentration of 20mmol.L -1 。
6. The method for preparing a ZIF-8-based phycocyanin molecularly imprinted drug carrier according to claim 1, wherein the centrifugation in the second step is performed at 5000rpm for 20min.
7. The method for preparing the phycocyanin molecularly imprinted drug carrier with ZIF-8 as matrix according to claim 1, wherein the volume ratio of DOX to PBS solution in the second step is 0.75g:1000mL; the volume ratio of the ZIF-8 to the PBS solution is 1 g/200 mL.
8. The preparation method of the phycocyanin molecular imprinting drug carrier with ZIF-8 as matrix according to claim 1, wherein the volume ratio of the ZIF-8/DOX to PBS solution in the third step is 1g:200mL; the volume ratio of the phycocyanin to the PBS solution is 1g to 1000mL; the volume ratio of the alpha-methacrylic acid to the PBS solution is 26g:10000mL.
9. The preparation method of the phycocyanin molecularly imprinted drug carrier with ZIF-8 as matrix according to claim 1, wherein the volume ratio of the N-isopropyl acrylamide to PBS solution in the third step is 34g:10000mL; the volume ratio of the polyethylene glycol 2000 to the PBS solution is 1g:200mL; the volume ratio of ammonium persulfate to PBS solution was 5.5g:10000mL.
10. The method for preparing the phycocyanin molecularly imprinted drug carrier with ZIF-8 as matrix according to claim 1, wherein MW=2000 Da of the PEG-FA in the third step; the volume ratio of the PEG-FA to the PBS solution is 1g to 100mL; said CaCO 3 The volume ratio to PBS solution was 1g:4000mL.
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| CN104492396A (en) * | 2014-12-10 | 2015-04-08 | 西南大学 | Targeted liposome based on protein molecular imprinting and preparation method thereof |
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| CN104492396A (en) * | 2014-12-10 | 2015-04-08 | 西南大学 | Targeted liposome based on protein molecular imprinting and preparation method thereof |
| CN110664778A (en) * | 2019-10-11 | 2020-01-10 | 上海交通大学医学院附属第九人民医院 | Composite microparticles, method for preparing same and use thereof as carrier in drug delivery |
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