CN112999365A - Preparation method of drug-loaded targeting magnetic nano composite particle - Google Patents
Preparation method of drug-loaded targeting magnetic nano composite particle Download PDFInfo
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- 239000003814 drug Substances 0.000 title claims abstract description 27
- 229940079593 drug Drugs 0.000 title claims abstract description 24
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 239000002245 particle Substances 0.000 title claims abstract description 17
- 230000008685 targeting Effects 0.000 title claims abstract description 16
- 239000002114 nanocomposite Substances 0.000 title claims abstract description 12
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 18
- 239000002105 nanoparticle Substances 0.000 claims abstract description 14
- 229920001184 polypeptide Polymers 0.000 claims abstract description 14
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 14
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims abstract description 12
- 239000002122 magnetic nanoparticle Substances 0.000 claims abstract description 7
- 229910052697 platinum Inorganic materials 0.000 claims abstract description 5
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims abstract description 4
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims abstract description 3
- GEPDYQSQVLXLEU-AATRIKPKSA-N methyl (e)-3-dimethoxyphosphoryloxybut-2-enoate Chemical compound COC(=O)\C=C(/C)OP(=O)(OC)OC GEPDYQSQVLXLEU-AATRIKPKSA-N 0.000 claims abstract 14
- 238000006243 chemical reaction Methods 0.000 claims description 34
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 18
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 18
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 17
- 239000000243 solution Substances 0.000 claims description 17
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 10
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 9
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- 238000007885 magnetic separation Methods 0.000 claims description 9
- 229960004316 cisplatin Drugs 0.000 claims description 7
- SZVJSHCCFOBDDC-UHFFFAOYSA-N iron(II,III) oxide Inorganic materials O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 claims description 7
- 238000003760 magnetic stirring Methods 0.000 claims description 6
- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical compound [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 claims description 6
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 claims description 5
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- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 claims description 3
- FALRKNHUBBKYCC-UHFFFAOYSA-N 2-(chloromethyl)pyridine-3-carbonitrile Chemical compound ClCC1=NC=CC=C1C#N FALRKNHUBBKYCC-UHFFFAOYSA-N 0.000 claims description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 3
- 239000000872 buffer Substances 0.000 claims description 3
- 239000007853 buffer solution Substances 0.000 claims description 3
- 229910052801 chlorine Inorganic materials 0.000 claims description 3
- 125000001309 chloro group Chemical group Cl* 0.000 claims description 3
- NQXWGWZJXJUMQB-UHFFFAOYSA-K iron trichloride hexahydrate Chemical compound O.O.O.O.O.O.[Cl-].Cl[Fe+]Cl NQXWGWZJXJUMQB-UHFFFAOYSA-K 0.000 claims description 3
- 239000011259 mixed solution Substances 0.000 claims description 3
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- 229910001961 silver nitrate Inorganic materials 0.000 claims description 3
- 239000002904 solvent Substances 0.000 claims description 3
- 229940014800 succinic anhydride Drugs 0.000 claims description 3
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- 238000010532 solid phase synthesis reaction Methods 0.000 claims 1
- 238000001291 vacuum drying Methods 0.000 claims 1
- 230000008499 blood brain barrier function Effects 0.000 abstract description 8
- 210000001218 blood-brain barrier Anatomy 0.000 abstract description 8
- 208000003174 Brain Neoplasms Diseases 0.000 abstract description 5
- 238000003384 imaging method Methods 0.000 abstract description 4
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- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 4
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- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 3
- WTFXARWRTYJXII-UHFFFAOYSA-N iron(2+);iron(3+);oxygen(2-) Chemical compound [O-2].[O-2].[O-2].[O-2].[Fe+2].[Fe+3].[Fe+3] WTFXARWRTYJXII-UHFFFAOYSA-N 0.000 description 3
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- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 description 1
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- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
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- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
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- A61K49/0032—Methine dyes, e.g. cyanine dyes
- A61K49/0034—Indocyanine green, i.e. ICG, cardiogreen
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Abstract
The invention belongs to the technical field of nano biological medicine, and particularly relates to a preparation method of drug-loaded targeting magnetic nano composite particles3O4Preparing nano particles CMDP/OTPBA-SPIONs carrying platinum drugs after preparing OTPBA-SPIONs; preparing an ICG labeled targeting polypeptide ANG-ICG; finally through amino and carboxyl groupsCoupling, namely modifying ANG-ICG on CMDP/OTPBA-SPIONs to obtain the multifunctional fluorescent targeting drug-loaded magnetic nanoparticles. The targeting polypeptide is used for increasing the ability of the medicine to pass through the blood brain barrier, and accurately positioning the brain tumor part through MRI imaging and near infrared fluorescence imaging, and meanwhile diagnosis and treatment of the brain tumor are realized.
Description
Technical Field
The invention belongs to the technical field of nano biomedicine, and relates to a preparation method of drug-loaded targeting magnetic nano composite particles, in particular to a preparation method of magnetic nanoparticles with coupled near-infrared dyes, targeting polypeptides and platinum anti-tumor drugs, so that multifunctional fluorescent targeting drug-loaded magnetic nanoparticles are obtained, the aim is to increase the ability of drugs to pass through a blood brain barrier through the targeting polypeptides, accurately position brain tumor parts through MRI imaging and near-infrared fluorescence imaging, and simultaneously realize diagnosis and treatment of brain tumors.
Background
The blood-brain barrier is an obstacle to protect the brain against the invasion of organic matter and foreign species, and more than 95% of drugs cannot cross the blood-brain barrier to enter the brain. Research shows that the nano-carrier can enter endothelial cells through endocytosis and then enter the brain through transcellular transfer, and the research shows that the research is greatly encouraging on the treatment of brain tumor diseases.
Iron oxide nanoparticles are a wide range of nanoparticles in the nano-family, and their unique magnetic properties can be applied to catalysis, data storage, bioseparation and MRI, which are of interest to many researchers. The superparamagnetic iron oxide nanometer can be used for MRI to guide drug transmission due to the excellent biocompatibility of the superparamagnetic iron oxide nanometer, and can also realize targeted transmission under the interference of an external magnetic field, so that the superparamagnetic iron oxide nanometer is widely applied to the field of biological medicines. However, one image has its own advantages and disadvantages, and it is very difficult to obtain accurate and reliable information of the disease location. Making up for their respective deficiencies accurate and reliable disease information can be obtained by the combined imaging tools, thus requiring combined imaging agents.
Disclosure of Invention
The invention aims to solve the technical problem of the application defects that the traditional platinum anticancer drugs are all chemical micromolecular drugs, have weaker targeting performance and obvious toxic and side effects and cannot pass through a blood brain barrier. Designing and synthesizing nanoparticles with proper size and biocompatibility to enable the drug small molecules to have the capability of passing through the blood brain barrier; on the basis, the surface of the nano material is further subjected to specific marking, so that the medicine can pass through a blood brain barrier and recognize the specificity of tumor cells, and the magnetic nano particles are subjected to near infrared fluorescence marking, so that nuclear magnetism and fluorescence visualization research and follow-up can be realized in clinic.
The technical scheme adopted by the invention for solving the technical problems is as follows:
preparing OTPBA;
succinic anhydride was added to methylene chloride under ice-bath conditions, and dissolved by strong magnetic stirring, and then a methylene chloride solution containing 3-aminopropyl-triethoxysilane was slowly added dropwise to the above solution over 1 hour, followed by transferring the reaction to room temperature and stopping the reaction after 5 hours. The solvent was removed using a rotary evaporator to give OTPBA.
(II) Fe3O4Preparing nano particles and carrying out surface modification to obtain OTPBA-SPIONs;
adding ferric trichloride hexahydrate and sodium hydroxide into ethylene glycol successively, heating to 70 ℃ under the condition of magnetic stirring until a uniformly mixed brown yellow solution is generated, transferring the uniformly mixed solution into a polytetrafluoroethylene reaction kettle, sealing, reacting at 240 ℃, stopping the reaction after 5 hours, and cooling the reaction kettle to room temperature. Solid product of black color (Fe)3O4) Collected by magnetic separation, then washed with water and ethanol, respectively, to remove unreacted reagents, and dried under vacuum at room temperature. Mixing Fe3O4The nanoparticles were ultrasonically dispersed in isopropanol (30mg/200mL), and then water and aqueous ammonia (mass fraction 30%) were added thereto, followed by dissolving 4-oxo-4- [3- (triethoxysilyl) propyl group in isopropanol]Amino-2-butyric acid (OTPBA) was slowly added dropwise to the above reaction system. The whole reaction is carried out at 50 ℃ under the ultrasonic condition, and the reaction is stopped after 3 h. The coated magnetic nanoparticles (OTPBA-SPIONs) were collected by magnetic separation, washed with water and ethanol to remove unreacted OTPBA, and the sample was vacuum dried at room temperature.
Preparing nano-particles CMDP/OTPBA-SPIONs carrying platinum drugs (CMDP);
cisplatin (DDP) and silver nitrate were dissolved in N, N-Dimethylformamide (DMF), and the reaction was stirred at room temperature in the dark for 12 hours. The reaction supernatant was centrifuged to remove the precipitate to give cisplatin CMDP deprived of one chlorine atom, which was present in the supernatant. The OTPBA-SPIONs are dispersed into a solution containing CMDP by an ultrasonic method, and the solution is stirred strongly for reaction for 48h under the condition of keeping out of the light. CMDP/OTPBA-SPIONs were obtained by magnetic separation and repeated washing with DMF and ethanol to remove unreacted CMDP. Whether the unreacted CMDP is completely washed away or not is judged by the Pt content of the supernatant obtained by ICP-MS test separation. The resulting sample was dried under vacuum at room temperature.
And (IV) preparing a near-infrared fluorescent dye ICG-labeled targeting polypeptide ANG-ICG, and modifying the ANG-ICG to CMDP/OTPBA-SPIONs through coupling of amino and carboxyl to obtain the ANG-ICG/CMDP/OTPBA-SPIONs.
Solid phase peptide Synthesis Using Fmoc chemistry Using Rink
The peptide sequence GGTFFYGGSRGKRNNFKTEEYGK(Mtt) -Resin was synthesized using a 5-fold excess of Fmoc protected amino acids and condensation reagents (HBTU/HOBt). The side chain Mtt-protected lysine was selectively deprotected with 1% TFA, and reacted with indocyanine green carboxylic acid (ICG-COOHCAS:181934-09-8), EDC, HOBt, anhydrous DMF to couple the side chain of lysine to ICG. Cleavage of the polypeptide from the resin with 95% TFA (and radical scavenger) gave NH2-GGTFFYGGSRGKRNNFKTEEYGK(ICG)-CONH2(ANG-ICG), precipitated with ether, the crude peptide obtained was analyzed by HPLC and isolated and purified, and the polypeptide was further verified by mass spectrometry.
CMDP/OTPBA-SPIONs particles were dispersed in buffer solution, EDC and NHS were added, stirred at room temperature for 30 minutes, ANG-ICG was added dropwise to the above buffer suspension (reaction protected from light), and stirred at 4 ℃ overnight. And then putting the reaction solution into a dialysis bag, and putting the bag into a large beaker filled with deionized water for dialysis so as to remove unreacted ICG, EDC and NHS. The sample in the bag is ANG-ICG/CMDP/OTPBA-SPIONs, and is stored at 4 ℃ in a closed light.
Has the advantages that:
according to the invention, brain glioma cells can be selectively identified by constructing a brain glioma targeting polypeptide, a near-infrared fluorescent dye ICG and cisplatin-modified magnetic ferroferric oxide nanoparticle, the polypeptide can be used as a targeting molecule to carry a nano complex to pass through a blood brain barrier and enter tumor cells, and the tumor part is accurately positioned through MRI imaging and near-infrared fluorescence imaging, so that the specific treatment of the brain glioma is realized.
Drawings
FIG. 1: a schematic of the preparation of OTPBA;
FIG. 2: fe3O4Scanning electron microscope photographs of (a);
FIG. 3: a preparation schematic diagram of OTPBA-SPIONs;
FIG. 4: scanning electron microscope photos of OTPBA-SPIONs;
FIG. 5: a schematic diagram of the preparation of CMDP/OTPBA-SPIONs;
FIG. 6: a preparation schematic diagram of ANG-ICG/CMDP/OTPBA-SPIONs;
FIG. 7: hydrated particle size plot of ANG-ICG/CMDP/OTPBA-SPIONs.
Detailed Description
The invention will be further described in the following examples, but it is to be understood that these examples are for illustrative purposes only and are not to be construed as limiting the practice of the invention.
Example 1
Preparation of fluorescent nano drug-loaded particles ANG-ICG/CMDP/OTPBA-SPIONs
Preparation of OTPBA
Succinic anhydride (1.0g) was added to methylene chloride (100mL) under ice-bath conditions, and dissolved by vigorous magnetic stirring, and then a solution of 3-aminopropyl-triethoxysilane (2.21g) in methylene chloride (10mL) was slowly added dropwise to the solution over 1h, after which the reaction apparatus was transferred to room temperature and the reaction was stopped after 5 h. The solvent was removed using a rotary evaporator to give the product.
Preparation of (di) OTPBA-SPIONs
Ferroferric oxide (Fe)3O4) And (3) preparing the nano particles.
Firstly, 1g of ferric chloride hexahydrate and 0.4g of sodium hydroxide are sequentially added into 30mL of ethylene glycol, the mixture is heated to 70 ℃ under the condition of magnetic stirring until a uniformly mixed brown yellow solution is generated, the uniformly mixed solution is transferred into a polytetrafluoroethylene reaction kettle to be sealed, the reaction is carried out at 240 ℃, the reaction is stopped after 5h, and the reaction kettle is cooled to room temperature. The black solid product was collected by magnetic separation, then washed with water and ethanol, respectively, to remove unreacted reagents, and dried under vacuum at room temperature.
Mixing Fe3O4The nanoparticles were ultrasonically dispersed in isopropanol (60mg/400mL), and then 4mL of water and 1mL of aqueous ammonia (mass fraction of 30%) were added thereto, followed by slowly dropping 0.5g of OTPBA dissolved in 50mL of isopropanol into the above reaction system. The whole reaction is carried out at 50 ℃ under the ultrasonic condition, and the reaction is stopped after 3 h. The coated magnetic nanoparticles (OTPBA-SPIONs) were collected by magnetic separation, washed with water and ethanol to remove unreacted OTPBA, and the sample was vacuum dried at room temperature.
Preparation of (tri) CMDP/OTPBA-SPIONs
100mg of cisplatin (DDP) and 56mg of silver nitrate were dissolved in 10mL of N, N-Dimethylformamide (DMF), and the reaction was stirred at room temperature with exclusion of light for 12 hours. The reaction supernatant was centrifuged to remove the precipitate to give cisplatin CMDP deprived of one chlorine atom, which was present in the supernatant.
And (d) dispersing the OTPBA-SPIONs obtained in the step (two) into the solution containing the CMDP by an ultrasonic method, and strongly stirring the solution for reaction for 48 hours in a dark condition. CMDP/OTPBA-SPIONs were obtained by magnetic separation and repeated washing with DMF and ethanol to remove unreacted CMDP. Whether the unreacted CMDP is completely washed away or not is judged by the Pt content of the supernatant obtained by ICP-MS test separation. The resulting sample was dried under vacuum at room temperature.
(IV) preparation of ANG-ICG/CMDP/OTPBA-SPIONs
Solid phase peptide Synthesis Using Fmoc chemistry Using Rink
The peptide sequence GGTFFYGGSRGKRNNFKTEEYGK(Mtt) -Resin was synthesized using a 5-fold excess of Fmoc protected amino acids and condensation reagents (HBTU/HOBt). Selectively deprotecting the side chain Mtt-protected lysine with 1% TFA, and addingReacting in ICG-COOH, EDC, HOBt and anhydrous DMF to couple the side chain of lysine with ICG. Cleavage of the polypeptide from the resin with 95% TFA (and radical scavenger) gave NH2-GGTFFYGGSRGKRNNFKTEEYGK(ICG)-CONH2(ANG-ICG), precipitated with ether, the crude peptide obtained was analyzed by HPLC and isolated and purified, and the polypeptide was further verified by mass spectrometry.
The CMDP/OTPBA-SPIONs particles were dispersed in a buffer solution (0.05M Na)2CO3/0.1M NaHCO3) EDC and NHS were added, and the mixture was stirred at room temperature for 30 minutes, and ANG-ICG (10mg/mL) was added dropwise to the above-mentioned buffer suspension (reaction was protected from light), and the mixture was stirred at 4 ℃ overnight. The reaction solution was placed in a dialysis bag (MWCO 30kDa) and dialyzed in a beaker of deionized water to remove unreacted ICG, EDC and NHS. The sample in the bag is ANG-ICG/CMDP/OTPBA-SPIONs, and is stored at 4 ℃ in a closed light.
In light of the foregoing description of the preferred embodiment of the present invention, it is to be understood that various changes and modifications may be made by those skilled in the art without departing from the spirit and scope of the invention. The technical scope of the present invention is not limited to the contents of the specification, and must be determined according to the scope of the claims.
Claims (8)
1. A preparation method of drug-loaded targeting magnetic nano composite particles is characterized by comprising the following steps:
(1) preparing OTPBA;
(2)Fe3O4preparing nano particles and carrying out surface modification to obtain OTPBA-SPIONs;
(3) preparing nano-particles CMDP/OTPBA-SPIONs carrying platinum drugs;
(4) preparing a near-infrared fluorescent dye ICG-labeled targeting polypeptide ANG-ICG, and modifying the ANG-ICG to CMDP/OTPBA-SPIONs through the coupling of amino and carboxyl to obtain the ANG-ICG/CMDP/OTPBA-SPIONs.
2. The method for preparing the drug-loaded targeted magnetic nanocomposite particle according to claim 1, wherein the method for preparing the OTPBA in the step (1) comprises the following steps: adding succinic anhydride into dichloromethane under ice bath condition, dissolving with strong magnetic stirring, slowly dripping dichloromethane solution containing 3-aminopropyl-triethoxysilane into the solution within 1h, then transferring to room temperature for reaction for 5h, and removing the solvent to obtain OTPBA.
3. The method for preparing the drug-loaded targeted magnetic nanocomposite particle according to claim 1, wherein the Fe in the step (2)3O4The preparation method of the nano particles comprises the following steps: sequentially adding ferric trichloride hexahydrate and sodium hydroxide into ethylene glycol, heating to 70 ℃ under magnetic stirring until a uniformly mixed brown yellow solution is generated, transferring the mixed solution into a polytetrafluoroethylene reaction kettle, sealing, reacting for 5 hours at 240 ℃, cooling the reaction kettle to room temperature, collecting black solids through magnetic separation, washing with water and ethanol respectively to remove unreacted reagents, and drying in vacuum at room temperature.
4. The preparation method of the drug-loaded targeted magnetic nanocomposite particle according to claim 1, wherein the method for obtaining OTPBA-SPIONs by surface modification in the step (2) comprises the following steps: mixing Fe3O4The nanoparticles were ultrasonically dispersed in isopropanol, and then water and aqueous ammonia were added thereto, followed by dissolving 4-oxo-4- [3- (triethoxysilyl) propyl group in isopropanol]Slowly dripping amino-2-butyric acid into the reaction system, reacting for 3h at 50 ℃ under the ultrasonic condition, collecting the wrapped magnetic nanoparticles (OTPBA-SPIONs) through magnetic separation, washing with water and ethanol respectively to remove unreacted OTPBA, and drying in vacuum at room temperature.
5. The preparation method of the drug-loaded targeted magnetic nanocomposite particle according to claim 1, wherein the preparation method of the nanoparticle CMDP/OTPBA-SPIONs in the step (3) comprises the following steps: dissolving cisplatin and silver nitrate in N, N-dimethylformamide, stirring at room temperature in dark place for 12h, centrifuging reaction clear liquid to remove precipitate to obtain cisplatin CMDP without one chlorine atom, allowing the cisplatin CMDP to exist in the clear liquid, ultrasonically dispersing OTPBA-SPIONs into a solution containing the CMDP, strongly stirring for reaction for 48h in dark place, removing unreacted CMDP from the CMDP/OTPBA-SPIONs through magnetic separation and repeated washing with DMF and ethanol, and vacuum drying at room temperature.
6. The method for preparing the drug-loaded targeted magnetic nanocomposite particle according to claim 1, wherein the method for preparing the near-infrared fluorescent dye ICG labeled targeted polypeptide ANG-ICG in the step (4) comprises the steps of firstly synthesizing a polypeptide sequence GGTFFYGGSRGKRNNFKTEEYGK(Mtt) -Resin by Fmoc chemical solid phase synthesis, and modifying to obtain NH2-GGTFFYGGSRGKRNNFKTEEYGK(ICG)-CONH2(ANG-ICG)。
7. The method for preparing the drug-loaded targeted magnetic nanocomposite particle according to claim 1, wherein the ANG-ICG/CMDP/OTPBA-SPIONs prepared in the step (4) are prepared by: dispersing CMDP/OTPBA-SPIONs particles in buffer solution, adding EDC and NHS, stirring at room temperature for 30 minutes, dropwise adding ANG-ICG into the buffer suspension, stirring at 4 ℃ in dark for overnight, dialyzing the reaction solution in a dialysis bag, and removing unreacted ICG, EDC and NHS to obtain the final product
ANG-ICG/CMDP/OTPBA-SPIONs。
8. A drug-loaded targeted magnetic nanocomposite particle made according to the method of any of claims 1-7.
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| XIAOYONG WANG等: "Functionalization of Platinum Complexes for Biomedical Applications", 《ACCOUNTS OF CHEMICAL RESEARCH》 * |
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