CN107837396B - Preparation method of multifunctional drug carrier integrating magnetic targeting, thermotherapy and chemotherapy - Google Patents
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Abstract
The invention relates to a preparation method of a multifunctional drug carrier integrating magnetic targeting, thermotherapy and chemotherapy, belonging to the field of drug carriers. The invention synthesizes MTX modified ferroferric oxide by a thermal decomposition method, and MTX-Fe is synthesized by the electrostatic attraction effect3O4And AuNPs are successfully assembled on MTX/LDHs to obtain the final product Fe3O4@ MTX/LDH-Au. The product has magnetic targeting function, thermotherapy and chemotherapy functions, and realizes multiple functions of drug carrier.
Description
Technical Field
The invention relates to a preparation method of a multifunctional drug carrier integrating magnetic targeting, thermotherapy and chemotherapy, belonging to the field of drug carriers.
Background
Magnetic drug carriers are a field with a wide development prospect, and the research on the preparation, performance and application of the magnetic drug carriers becomes a hot point of research. In preparation, the magnetic nano material is generally a core-shell type microsphere, and a plurality of researchers have searched the core-shell type magnetic nano material. MgFe prepared by seguing et al2O4@ IBU (ibuprofen)/LDH magnetic composite, Fe3O4@ DFUR (deoxyfluorouridine)/LDH complex and Fe3O4@SiO2@ LDH microspheres; fe is synthesized by professor of Houwangshi3O4@ LDH complex; shijialin researchers and the like synthesize Fe3O4@SiO2@ LDH core-shell structure, and the like. Under the action of an external magnetic field, the medicine carrying system can carry medicines to a target part, so that the targeting property is improved.
The research on the drug carrier tends to cooperate in a plurality of functions.
Disclosure of Invention
According to the defects of the prior art, the technical problems to be solved by the invention are as follows: the product has magnetic targeting function, thermotherapy and chemotherapy functions, and realizes multiple functions of the drug carrier.
The invention synthesizes MTX modified ferroferric oxide by a thermal decomposition method, and MTX-Fe is synthesized by the electrostatic attraction effect3O4And AuNPs are successfully assembled on MTX/LDHs to obtain the final product Fe3O4@MTX/LDH-Au。
The technical scheme adopted by the invention for solving the technical problems is as follows: the preparation method of the multifunctional drug carrier integrating magnetic targeting, thermotherapy and chemotherapy comprises the following steps:
(1) synthesis of Fe by thermal decomposition3O4-MTX：
FeCl is added3.6H2Dissolving O in ethylene glycol, adding NaAc, polyethylene glycol and MTX into the solution, stirring, transferring to a high-pressure reaction kettle, placing the high-pressure reaction kettle into an oven after the reaction is finished, and performing hydrothermal treatment to obtain Fe3O4-a MTX nanocomposite;
(2)Fe3O4synthesis of @ MTX/LDH nanomaterial:
preparation of solution A: the newly synthesized Fe3O4-adding the MTX nanocomposite to an aqueous alcohol solution, dissolving MTX in NH3·H2O, transferring the mixture into the alcohol-water solution, wherein the mixed solution is marked as solution A;
preparation of mixed salt solution B: mixing Mg (NO)3)3·6H2O and Al (NO)3)3·9H2Dissolving O in 50mL of alcohol-water solution to prepare a mixed salt solution B,
transferring the solution A into a three-neck flask, continuously introducing nitrogen, dropwise adding the mixed salt solution B into the solution A, adjusting the pH value of the mixed solution to 9.5 in the dropwise adding process, keeping the temperature at 60 ℃, magnetically stirring, and continuously introducing N2And stopping the reaction after 1h to obtain a product Fe3O4@MTX/LDH；
(3) The synthesis of the magnetic targeting, thermotherapy and chemotherapy integrated multifunctional drug carrier:
adopting seed growth method to synthesize AuNPs, adding sodium citrate solution into round-bottom flask, and placing in oil bathHeated to boiling and then added HAuCl dropwise4Changing the solution from yellow to wine red until the color is not changed any more to form gold seeds; cooling the gold seeds to 90 ℃, and adding a sodium citrate solution and HAuCl4Stirring and reacting to obtain gold nanoparticles with the particle size of 30 nm;
centrifuging and washing newly synthesized AuNPs, dispersing into distilled water again, and adding into Fe3O4Stirring at room temperature in the @ MTX/LDH nano compound to obtain the final product Fe3O4@MTX/LDH-Au。
The method adopts a thermal decomposition method to synthesize Fe3O4-MTX：
1.35g FeCl was weighed3.6H2Dissolving O in 40mL of ethylene glycol, respectively weighing 3.6g of NaAc, 1.0g of polyethylene glycol and 30mg of MTX, adding the NaAc, the polyethylene glycol and the MTX into the solution, stirring the solution at room temperature for 30min, transferring the solution into a 50mL high-pressure reaction kettle (with the pressure of 15-30MPa) for reaction, placing the reaction product into an oven after the reaction is finished, and carrying out hydrothermal treatment at 180 ℃ for 24h to obtain Fe3O4-MTX nanocomposite.
Said Fe3O4Synthesis of @ MTX/LDH nanomaterial:
preparation of solution A: 50mg of newly synthesized Fe was accurately weighed3O4MTX nanocomposite was added to 50mL of an aqueous alcohol solution (1: 3 volume ratio of ethanol to water), 0.0341g of MTX was dissolved in 8mL of 10% NH3·H2O, transferring the mixed solution into the solution, wherein the mixed solution is marked as solution A;
preparation of mixed salt solution B: weighing 0.07692g Mg (NO)3)3·6H2O and 0.05627g Al (NO)3)3·9H2Dissolving the mixed salt solution in 50mL of alcohol-water solution (the volume ratio of ethanol to water is 1:3) to prepare a mixed salt solution B,
transferring the solution A into a three-neck flask, continuously introducing nitrogen to protect ferroferric oxide in the solution, then dropwise adding a mixed salt solution B into the solution A, adjusting the pH value of the mixed solution to 9.5 by using ammonia water with the mass fraction of 10% in the dropwise adding process, keeping the temperature at 60 ℃, magnetically stirring and continuously introducingN2And stopping the reaction after 1h to obtain a product Fe3O4@MTX/LDH。
The synthesis of the magnetic-collecting targeting, thermotherapy and chemotherapy integrated multifunctional drug carrier comprises the following steps:
au NPs are synthesized by adopting a seed growth method, 150mL of 0.0022mol/L sodium citrate solution is added into a round-bottom flask, the round-bottom flask is heated to boiling in oil bath at 137 ℃, and then 1mL of 0.025mol/L HAuCl is added dropwise4Changing the solution from yellow to wine red until the color is not changed any more to form gold seeds;
the gold seeds are cooled to 90 ℃, and 3mL of 0.06mol/L sodium citrate solution and 1mL of 0.025mol/L HAuCl are rapidly added4Magnetically stirring for reaction for 1h to obtain gold nanoparticles with the particle size of 30 nm;
20mL of freshly synthesized Au NPs were washed by centrifugation, redispersed in 20mL of distilled water, and added to 10mL of 10mg/mL Fe3O4Stirring for 5 hours at room temperature in the @ MTX/LDH nano compound to obtain the final product Fe3O4@MTX/LDH-Au。
The volume ratio of alcohol to water in the alcohol-water solution is 1:3 (alcohol Water the alcohol here is ethanol).
The invention has the beneficial effects that:
1. the invention synthesizes MTX modified ferroferric oxide by a thermal decomposition method, and MTX-Fe is synthesized by the electrostatic attraction effect3O4And Au NPs are successfully assembled on the MTX/LDHs to obtain a final product Fe3O4@ MTX/LDH-Au. The product has magnetic targeting function, thermotherapy and chemotherapy functions, and realizes multiple functions of drug carrier;
2. fe worth of the invention3O4The release of the @ MTX/LDH-Au complex in PBS is mainly divided into two stages, the release rate is faster at the first 100min, and the release rate is slower at 100-500min, which is similar to the drug release rule of MTX/LDH. We also believe that the first stage is primarily Fe adsorbed3O4The release of the drug from the surface of @ MTX/LDH-Au, whereas the second phase of drug release is mainly caused by ion exchange of the drug molecules in the interlayer with phosphate anions. In summary, Fe3O4The @ MTX/LDH-Au compound has better slow release performance.
Drawings
FIG. 1 is Fe3O4-MTX、Fe3O4@ MTX/LDH and Fe3O4TEM image of @ MTX/LDH-Au;
FIG. 2 shows Fe in sample3O4-MTX、Fe3O4@MTX/LDH、Fe3O4XRD patterns of @ MTX/LDH-Au and Au-MTX;
FIG. 3 shows MTX and Fe samples3O4-MTX、Fe3O4@ MTX/LDH and Fe3O4FTIR profile of @ MTX/LDH-Au;
FIG. 4 is Fe3O4The sustained release curve chart of the @ MTX/LDH-Au nano compound;
wherein the (A, B) sample in FIG. 1 is Fe3O4-TEM images of MTX; (C, D) sample is Fe3O4TEM image of @ MTX/LDH; (E, F) sample is Fe3O4TEM image of @ MTX/LDH-Au.
Detailed Description
Embodiments of the invention are further described below with reference to the accompanying drawings:
example 1
As shown in figures 1-4, the invention relates to a preparation method of a multifunctional drug carrier integrating magnetic targeting, thermotherapy and chemotherapy, which comprises the following steps:
(1) synthesis of Fe by thermal decomposition3O4-MTX；
(2)Fe3O4Synthesizing a @ MTX/LDH nano material;
(3) the synthesis of the multifunctional drug carrier integrating magnetic targeting, thermotherapy and chemotherapy.
The method adopts a thermal decomposition method to synthesize Fe3O4-MTX：
1.35g FeCl was weighed3.6H2Dissolving O in 40mL of ethylene glycol, respectively weighing 3.6g of NaAc, 1.0g of polyethylene glycol and 30mg of MTX, adding into the solution, stirring at room temperature for 30min, transferring into a 50mL high-pressure reaction kettle (under the pressure of 15-30MPa) for reaction, and reactingAfter the completion of the preparation, the mixture is put into an oven and is subjected to hydrothermal treatment for 24 hours at 180 ℃ to prepare Fe3O4-MTX nanocomposite.
Said Fe3O4Synthesis of @ MTX/LDH nanomaterial:
preparation of solution A: 50mg of newly synthesized Fe was accurately weighed3O4MTX nanocomposite was added to 50mL of an aqueous alcohol solution (ethanol/water volume ratio 1:3), 0.0341g of MTX was dissolved in 8mL of 10% NH3·H2O, transferring the mixed solution into the solution, wherein the mixed solution is marked as solution A;
preparation of mixed salt solution B: weighing 0.07692g Mg (NO)3)3·6H2O and 0.05627g Al (NO)3)3·9H2O, dissolving the mixed salt solution in 50mL of alcohol-water solution (the volume ratio of ethanol to water is 1:3) to prepare a mixed salt solution B,
transferring the solution A into a three-neck flask, continuously introducing nitrogen to protect ferroferric oxide in the solution, then dropwise adding a mixed salt solution B into the solution A, adjusting the pH value of the mixed solution to 9.5 by using ammonia water with the mass fraction of 10% in the dropwise adding process, keeping the temperature at 60 ℃, magnetically stirring, and continuously introducing N2And stopping the reaction after 1h to obtain a product Fe3O4@MTX/LDH。
The synthesis of the magnetic-collecting targeting, thermotherapy and chemotherapy integrated multifunctional drug carrier comprises the following steps:
au NPs are synthesized by adopting a seed growth method, 150mL of 0.0022mol/L sodium citrate solution is added into a round-bottom flask, the round-bottom flask is heated to boiling in oil bath at 137 ℃, and then 1mL of 0.025mol/L HAuCl is added dropwise4Changing the solution from yellow to wine red until the color is not changed any more to form gold seeds;
the gold seeds are cooled to 90 ℃, and 3mL of 0.06mol/L sodium citrate solution and 1mL of 0.025mol/L HAuCl are rapidly added4Magnetically stirring for reaction for 1h to obtain gold nanoparticles with the particle size of 30 nm;
20mL of freshly synthesized Au NPs were washed by centrifugation, redispersed in 20mL of distilled water, and added to 10mL of 10mg/mL Fe3O4@ MTX/LDH nano-scaleStirring the mixture for 5 hours at room temperature to obtain a final product Fe3O4@MTX/LDH-Au。
The volume ratio of alcohol to water in the alcohol-water solution is 1:3 (alcohol Water the alcohol here is ethanol).
As can be seen from FIG. 1, Fe3O4The MTX distribution is relatively uniform and the particle size is uniform. Measuring Fe3O4Zeta potential of MTX is-40.5 mV, which has strong stability, and magnesium aluminum salt solution, MTX solution and Fe are mixed by electrostatic action3O4Mixing in Fe3O4Growing in situ to generate Fe with a core-shell structure3O4@ MTX/LDH nanocomposite (FIGS. 1C-D), in which it is seen that a significant LDH thin shell is wrapped in Fe3O4Surface of (2) proves Fe3O4The presence of the @ MTX/LDH nanocomposite structure. Mixing Fe3O4Mixing and stirring @ MTX/LDH and Au NPs with a certain volume to obtain Fe3O4@ MTX/LDH-Au nanocomposites (FIGS. 1E-F), Au NPs successfully adsorbed to Fe by electrostatic attraction3O4@ MTX/LDH surface.
Sample Fe3O4-MTX、Fe3O4@MTX/LDH、Fe3O4The XRD patterns of @ MTX/LDH-Au and Au-MTX are shown in FIG. 2, sample Fe3O4MTX shows strong characteristic peaks at 30.1 °, 35.5 °, 43.1 °, 53.4 °, 57.0 ° and 62.6 °, which corresponds to Fe3O4Diffraction peaks of (220), (311), (400), (422), (511), and (440) crystal planes of the inverse cubic spinel structure. In Fe3O4@ MTX/LDH, except for Fe3O4Besides the characteristic peak of MTX, there is also a diffraction peak of LDH, but the intensity is not high. For Au-MTX NPs, the strong peaks appearing at 38 °, 44.6 °, 64.8 ° and 77.8 ° 2 θ correspond to the characteristic diffraction peaks of the (111), (200), (220) and (311) crystal planes of Au NPs. And in Fe3O4In the sample of @ MTX/LDH-Au, Fe3O4Characteristic diffraction peaks of MTX, Au NPs and LDH all exist. The small part of the upper right hand corner of the XRD pattern is sample Fe3O4@ MTX/LDH-Au XRD at 2 theta of 2-20 deg.,the presence of the (003) and (006) diffraction peaks of LDH, together with NO, is evident in this figure3The (003) diffraction peak of the sample shifted in the low angle direction compared to the LDHs (003 characteristic diffraction peak corresponding to a 2 theta angle of 10.92 °), indicating successful intercalation of the drug MTX into the LDHs interlayer. To sum up, the sample Fe3O4@ MTX/LDH-Au has been synthesized successfully.
FIG. 3 shows MTX and Fe samples3O4-MTX、Fe3O4@ MTX/LDH and Fe3O4FTIR profile of @ MTX/LDH-Au. Sample Fe3O4-MTX、Fe3O4@ MTX/LDH and Fe3O4@ MTX/LDH-Au at 575cm-1The characteristic absorption peak of Fe-O appears, which indicates that Fe3O4Are all present in the above-mentioned substances. Comparing the samples, it can be found that the wave number of the four samples is 1614cm-1And 1100cm-1The two absorption peaks respectively correspond to COO in MTX-The antisymmetric vibration peak and the stretching vibration peak of primary amine and tertiary amine C-N bond in MTX aromatic ring prove that MTX is successfully loaded on Fe3O4-MTX、Fe3O4@ MTX/LDH and Fe3O4@ MTX/LDH-Au nanoparticle surface.
Fe3O4The drug loading of the @ MTX/LDH-Au nano-composite is 38.2%, and Fe is explored in PBS buffer solution with pH 7.4 at 37 DEG C3O4The sustained release performance of @ MTX/LDH-Au (figure 4) shows that the compound has very obvious sustained release effect, the sustained release curve is stable, and the phenomenon of burst release of the drug does not occur. Fe3O4The release of the @ MTX/LDH-Au complex in PBS is mainly divided into two stages, the release rate is faster at the first 100min, and the release rate is slower at 100-500min, which is similar to the drug release rule of MTX/LDH. We also believe that the first stage is primarily Fe adsorbed3O4The release of the drug from the surface of @ MTX/LDH-Au, while the second phase of drug release is mainly caused by ion exchange of the drug molecules with phosphate anions between the layers[106]. In conclusion, Fe3O4The @ MTX/LDH-Au compound has better slow release performance.
Claims (2)
1. A preparation method of a multifunctional drug carrier integrating magnetic targeting, thermotherapy and chemotherapy is characterized by comprising the following steps:
(1) synthesis of Fe by thermal decomposition3O4-MTX：
FeCl is added3 .6H2Dissolving O in ethylene glycol, adding NaAc, polyethylene glycol and MTX into the solution, stirring, transferring to a high-pressure reaction kettle, placing the high-pressure reaction kettle into an oven after the reaction is finished, and performing hydrothermal treatment to obtain Fe3O4-a MTX nanocomposite;
(2)Fe3O4synthesis of @ MTX/LDH nanomaterial:
preparation of solution A: the newly synthesized Fe3O4-adding the MTX nanocomposite to an aqueous alcohol solution, dissolving MTX in NH3·H2O, transferring the mixture into the alcohol-water solution, wherein the mixed solution is marked as solution A;
preparation of mixed salt solution B: mixing Mg (NO)3)3·6H2O and Al (NO)3)3·9H2Dissolving O in 50mL of alcohol-water solution to prepare a mixed salt solution B,
transferring the solution A into a three-neck flask, continuously introducing nitrogen, dropwise adding the mixed salt solution B into the solution A, adjusting the pH value of the mixed solution to 9.5 in the dropwise adding process, keeping the temperature at 60 ℃, magnetically stirring, and continuously introducing N2And stopping the reaction after 1h to obtain a product Fe3O4@MTX/LDH；
(3) The synthesis of the magnetic targeting, thermotherapy and chemotherapy integrated multifunctional drug carrier:
synthesizing AuNPs by adopting a seed growth method: the sodium citrate solution was added to the round bottom flask, heated to boiling in an oil bath, and then HAuCl was added dropwise4Changing the solution from yellow to wine red until the color is not changed any more to form gold seeds; cooling the gold seeds to 90 ℃, and adding a sodium citrate solution and HAuCl4Stirring and reacting to obtain gold nanoparticles with the particle size of 30 nm;
centrifuging and washing newly synthesized AuNPs, dispersing into distilled water again, and adding into Fe3O4Stirring at room temperature in the @ MTX/LDH nano compound to obtain the final product Fe3O4@MTX/LDH-Au；
The method adopts a thermal decomposition method to synthesize Fe3O4-MTX：
1.35g FeCl was weighed3 .6H2Dissolving O in 40mL of ethylene glycol, respectively weighing 3.6g of NaAc, 1.0g of polyethylene glycol and 30mg of MTX, adding the NaAc, the polyethylene glycol and the MTX into the solution, stirring the solution at room temperature for 30min, transferring the solution into a 50mL high-pressure reaction kettle for reaction, keeping the pressure at 15-30MPa, putting the reaction product into an oven after the reaction is finished, and carrying out hydrothermal treatment at 180 ℃ for 24h to obtain Fe3O4-a MTX nanocomposite;
said Fe3O4Synthesis of @ MTX/LDH nanomaterial:
preparation of solution A: 50mg of newly synthesized Fe was accurately weighed3O4Adding the MTX nano-composite into 50mL of alcohol-water solution, wherein the volume ratio of ethanol to water is 1:3, and dissolving 0.0341g of MTX into 8mL of 10% NH3·H2O, transferring the mixed solution into the solution, wherein the mixed solution is marked as solution A;
preparation of mixed salt solution B: weighing 0.07692g Mg (NO)3)3·6H2O and 0.05627g Al (NO)3)3·9H2Dissolving the mixed salt solution B in 50mL of alcohol-water solution to prepare a mixed salt solution B, wherein the volume ratio of ethanol to water is 1: 3;
transferring the solution A into a three-neck flask, continuously introducing nitrogen to protect ferroferric oxide in the solution, then dropwise adding a mixed salt solution B into the solution A, adjusting the pH value of the mixed solution to 9.5 by using ammonia water with the mass fraction of 10% in the dropwise adding process, keeping the temperature at 60 ℃, magnetically stirring, and continuously introducing N2And stopping the reaction after 1h to obtain a product Fe3O4@MTX/LDH。
2. The preparation method of the magnetic collection targeting, thermotherapy and chemotherapy integrated multifunctional drug carrier according to claim 1, wherein the synthesis of the magnetic collection targeting, thermotherapy and chemotherapy integrated multifunctional drug carrier comprises:
synthesizing AuNPs by adopting a seed growth method: 150mL of 0.0022mol/L sodium citrate solution was added to a round bottom flask, heated to boiling in a 137 ℃ oil bath, and 1mL of 0.025mol/L HAuCl was added dropwise4Changing the solution from yellow to wine red until the color is not changed any more to form gold seeds;
the gold seeds are cooled to 90 ℃, and 3mL of 0.06mol/L sodium citrate solution and 1mL of 0.025mol/L HAuCl are rapidly added4Magnetically stirring for reaction for 1h to obtain gold nanoparticles with the particle size of 30 nm;
20mL of freshly synthesized Au NPs were washed by centrifugation, redispersed in 20mL of distilled water, and added to 10mL of 10mg/mL Fe3O4Stirring for 5 hours at room temperature in the @ MTX/LDH nano compound to obtain the final product Fe3O4@MTX/LDH-Au。
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| CN106865622A (en) * | 2017-02-16 | 2017-06-20 | 吴晓红 | A kind of Fe3O4The synthetic method of@LDH composite nano materials |
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| CN106865622A (en) * | 2017-02-16 | 2017-06-20 | 吴晓红 | A kind of Fe3O4The synthetic method of@LDH composite nano materials |
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| "A Novel Platform Designed by Au Core/Inorganic Shell Structure Conjugated onto MTX/LDH for Chemo-photothermal Therapy";De-Ying Tian等;《Int. J. Pharm.》;20161231(第505期);摘要，第5页 * |
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