CN114469897A - Hectorite composite delivery material for targeted therapy of myocardial infarction and preparation and application thereof - Google Patents
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Abstract
The invention relates to a hectorite composite delivery material for targeted therapy of myocardial infarction and preparation and application thereof. Compared with the prior art, the invention has the characteristics of simple preparation method, controllable alkali amount and alkali strength, high drug loading capacity and strong biological safety, the drug has high myocardial targeting property, can be locally and accurately released, has longer elimination half-life period than the traditional drug, has higher bioavailability of the drug, and provides a controllable, slow-release and efficient multifunctional composite platform for drug delivery of myocardial infarction.
Description
Technical Field
The invention belongs to the technical field of nano biomaterials, and relates to a hectorite composite delivery material for targeted therapy of myocardial infarction, and a preparation method and application thereof.
Background
Myocardial infarction is a big killer threatening human life health, and although coronary bypass and stent intervention are widely performed clinically, the death rate caused by myocardial infarction is still high. Therefore, the search for new strategies for treating myocardial infarction is urgent. Research shows that microRNA (miRNA) plays an important regulation role in the pathogenesis process of myocardial infarction. Various miRNAs such as miR-1, miR-133a, miR-92a and the like play an important role in pathological processes such as apoptosis, oxidative stress, angiogenesis and the like in myocardial infarction tissues, and are expected to become effective gene therapy means for myocardial infarction. A safe and efficient delivery system is selected, and the premise that miRNA is carried to local lesion of myocardial infarction to play a therapeutic role is provided.
Compared with the traditional viral vector, the miRNA is delivered by adopting the nano-vector, so that the biological safety and the in vitro targeting performance are good. However, the in vivo delivery efficiency of the nano-miRNA is not ideal because miRNA has a short half-life in blood circulation and does not have active targeting to infarcted myocardium.
Disclosure of Invention
The invention aims to provide a safe and high-load sustained-release type lithium-saponite-based nano delivery material targeting local myocardial infarction and preparation and application thereof.
The purpose of the invention can be realized by the following technical scheme:
the composite hectorite delivery material for targeted therapy of myocardial infarction comprises hectorite loaded with microRNA and targeted specific cell membranes coated outside.
Further, the hectorite comprises one or more of RD hectorite, RDs hectorite, XLG hectorite, XLS hectorite or S482 hectorite, the microRNA comprises one or more of miR-1, miR-204, miR-133a or miR-92a, and the target specific cell membrane is one of platelet membrane, macrophage membrane or exosome.
Furthermore, in the composite delivery material, the mass percentage of the microRNA is 0.01-20%, and the mass percentage of the targeted specific cell membrane is 0.01-20%.
Preferably, the laponite composite delivery material is formulated such that the proportion of solids to the mass of the entire liquid (mass ratio) is 0.01-20%.
Preferably, the composite delivery material further comprises a solvent, the mass ratio of the solvent to the hectorite being 4-1000: 1.
The preparation method of the hectorite composite delivery material for targeted therapy of myocardial infarction comprises the following steps:
1) mixing the microRNA solution with hectorite, carrying out exchange adsorption reaction, and separating to obtain a slow-release microRNA-hectorite composite material;
2) dispersing the slow-release microRNA-hectorite composite material in a solvent, adding a target specific cell membrane, carrying out adsorption and coating reaction under stirring, and separating to obtain the hectorite composite delivery material for targeted treatment of myocardial infarction.
Further, in the step 1), the concentration of the microRNA solution is 0.5 wt% to 5 wt%, the temperature of the exchange adsorption reaction is 0 to 50 ℃, and the time is 0.1 to 20 hours.
Further, in the step 2), the solvent is water, the temperature of the adsorption and coating reaction is 0-50 ℃, and the time is 1-48 h.
Further, in the step 2), a product obtained by separation after the adsorption and coating reaction is added into a solvent to prepare a solution.
In the preparation process, a centrifugal method is adopted for separation, and an ice bath and water bath mode is adopted for temperature control.
The application of the composite delivery material in preparing a medicament for targeted therapy of myocardial infarction. The composite delivery material plays an important role in modulating local apoptosis of ischemic myocardium, oxidative stress and endothelial neovascularization, prolongs elimination half-life period compared with the traditional medicine, and improves bioavailability of the medicine. The nanoparticles coated with the specific cell membranes show stronger myocardial targeting on in-vivo imaging, are sensitive to weakly acidic pH around myocardial infarction, and can be precisely released in a targeted manner locally.
The drug for targeted therapy of myocardial infarction contains the composite delivery material.
Laponite is an artificially synthesized silicate having a specific layered structure. The hectorite nanolayers have an anisotropic charge distribution: the edge is positively charged and the outer surface is negatively charged. The hectorite has good ion exchange capacity (50-150 mmol/100g at pH 6-13) and large specific surface area (350 m)2In terms of/g). When the hectorite crystals are dispersed in water, the hectorite crystals are close to each other, and surface charges can repel each other, so that the hectorite crystals can be uniformly dispersed in the water, and the hydrogel or the sol which flows in Newtonian mode and low viscosity is easy to form, and has adjustable thixotropy and rheological properties.
The invention adopts porous hectorite nano clay material with large surface area, high biocompatibility and good water dispersibility as a carrier, loads microRNA which plays an important role in modulating local apoptosis of ischemic myocardium, oxidative stress and endothelial neovascularization, then uses biomembrane technology to wrap specific cell membrane to achieve targeted effect on infarcted myocardium local, and utilizes the pH sensitivity characteristic of hectorite to accurately release the medicine. The material has the characteristics of simple preparation method, controllable alkali amount and alkali strength, high drug loading amount and strong biological safety, the drug has high myocardial targeting property, can be locally and accurately released, prolongs the elimination half-life period compared with the traditional drug, improves the bioavailability of the drug, and provides a controllable, slow-release and efficient multifunctional composite platform for drug delivery of myocardial infarction.
Compared with the prior art, the invention has the following characteristics:
1) the nano-carrier hectorite applied by the invention has good ion exchange capacity and large specific surface area, and when the hectorite is dispersed in water, the hectorite crystals are close to each other, and surface charges can repel each other, so that the hectorite crystals can be uniformly dispersed in the water, a Newtonian and low-viscosity flowing hydrogel or sol is easily formed, and the nano-carrier hectorite has adjustable thixotropy and rheological property, and can be used for synthesizing various nano-composite materials according to the design requirements of nano-composites.
2) The hectorite composite delivery material disclosed by the invention is stable in structure, is a potential excellent carrier for encapsulating miRNA, can overcome particle agglutination reaction, shields the identification of an endothelial reticular system, can play a role in systemic circulation for a long time, and improves the miRNA delivery rate.
3) The hectorite composite delivery material has the characteristics of in vivo degradability and good biocompatibility, is not easy to be removed by an autoimmune system in the miRNA delivery process, is easy to penetrate blood vessels without causing vascular endothelial injury, protects the medicament from enzymatic degradation, has high biosafety concentration, and can improve the miRNA delivery load capacity and the myocardial local medicament aggregation concentration.
4) The hectorite composite delivery material disclosed by the invention utilizes a cell membrane bionic technology to apply platelet membranes, macrophage membranes or exosomes to coating of the nanoparticles, so that the nanoparticles are directionally migrated to local ischemic myocardium along with the targeted delivery property of specific receptors or membrane proteins on the cell membranes, and the hectorite composite delivery material has the accurate targeting property for treating myocardial infarction and improves the action efficiency.
5) The hectorite composite delivery material plays an important role in modulating local apoptosis of ischemic cardiac muscle, oxidative stress and endothelial neovascularization.
6) The hectorite composite delivery material disclosed by the invention can enable the medicine to be slowly released under the local weak acid condition of myocardial infarction by utilizing the layered structure, high load capacity and pH value dependence of the hectorite, so that the half life of the medicine is prolonged compared with that of the traditional medicine, and the bioavailability of the medicine is improved.
Drawings
FIG. 1 is a graph of the results of fluorescence microscopy of myocardial cells assessing nanoparticle uptake and phagocytosis in the examples;
FIG. 2 flow cytometry evaluation of nanoparticle pairs H in examples2O2Results of the apoptotic effect of cardiomyocytes under stimulation are shown, wherein a is the cardiomyocytes under normal environment, and b is H2O2The stimulated myocardial cells, c are the myocardial cells added with the hectorite nano-composite AL @ M-2;
fig. 3 is a graph of the results of ex vivo imaging evaluation of the targeting effect of nanoparticles on myocardial tissue in examples.
Detailed Description
The invention is described in detail below with reference to the figures and specific embodiments. The present embodiment is implemented on the premise of the technical solution of the present invention, and a detailed implementation manner and a specific operation process are given, but the scope of the present invention is not limited to the following embodiments.
The invention provides a hectorite composite delivery material for targeted therapy of myocardial infarction, which comprises a hectorite loaded with microRNA and a targeted specific cell membrane coated outside the hectorite.
Wherein the hectorite comprises one or more of RD hectorite, RDS hectorite, XLG hectorite, XLS hectorite or S482 hectorite, the microRNA comprises one or more of miR-1, miR-204, miR-133a or miR-92a, and the targeted specific cell membrane is one of platelet membrane, macrophage membrane or exosome. In the composite delivery material, the mass percentage of the microRNA is 0.01-20%, and the mass percentage of the targeted specific cell membrane is 0.01-20%.
Preferably, the composite delivery material further comprises a solvent, the mass ratio of the solvent to the hectorite being 4-1000: 1.
The invention also provides a preparation method of the composite delivery material, which comprises the following steps:
1) mixing the microRNA solution with hectorite, carrying out exchange adsorption reaction, and separating to obtain a slow-release microRNA-hectorite composite material;
2) dispersing the sustained-release microRNA-hectorite composite material in a solvent, adding a target specific cell membrane, carrying out adsorption and wrapping reaction under stirring, and separating to obtain the hectorite composite delivery material for targeted treatment of myocardial infarction.
In the step 1), the concentration of the microRNA solution is 0.5 wt% -5 wt%, the temperature of the exchange adsorption reaction is 0-50 ℃, and the time is 0.1-20 h.
In the step 2), the solvent is water, the temperature of the adsorption and coating reaction is 0-50 ℃, and the time is 1-48 h. Preferably, a product obtained by separation after the adsorption coating reaction is further added into a solvent to prepare a solution.
The invention also provides application of the composite delivery material in preparation of a medicament for targeted treatment of myocardial infarction.
The invention further provides a medicine for targeted therapy of myocardial infarction, which contains the composite delivery material.
Example 1:
mixing a microRNA (anti-miR-1 antisense oligonucleotide, AMO-1 for short) solution with the concentration of 1.0 wt% with RD type hectorite (RD-Lp) according to the liquid-solid ratio of 50:1, performing exchange adsorption for 8.0h at 50 ℃, centrifuging to obtain a solid product, washing for multiple times, and centrifuging to obtain the sustained-release [ AMO-1/RD-Lp ] -1 composite delivery material (AL-1 for short). Adding the obtained composite material into a 1.0 wt% Platelet Membrane (PM) aqueous solution (liquid-solid ratio is 50:1), stirring at 25 ℃, adsorbing and wrapping for 10.0h, centrifuging, and preparing into a 1 wt% solution to obtain the hectorite composite delivery material [ AMO-1/RD-Lp @ PM ] -1 (AL @ M-1 for short) for targeted treatment of myocardial infarction.
The phagocytic uptake efficiency of the myocardial cells on AL-1 and AL @ M-1 and the distribution condition in the cells are observed through the localization of a fluorescence microscope.
1) Preparing Fluorescein Isothiocyanate (FITC) labeled Nanoparticles (NPs) AL-1 and AL @ M-1 in advance: respectively dispersing 100mg of AL-1 solution and 100mg of AL @ M-1 solution in 8mL of deionized water, adding 3mL of FITC ethanol solution (1mL/mg), stirring for 6h in the dark, centrifuging at 4000g, collecting nanoparticles, washing for 3 times, filtering with a 0.45-micron filter membrane, and sampling for quantitative use.
2) According to the cell concentration of 5 ten thousand per hole, the extracted primary newborn rat myocardial cells (NRCMs) are planted in a 24-hole plate, and the subsequent experiment is carried out after the cells are completely attached to the wall;
3) using a low-sugar medium (DMEM) to dilute AL-1 and AL @ M-1 to 200 mu g/mL for later use;
4) the cells of each group were modeled and dosed according to the described protocol and incubated for 3 h;
5) after 3h, the medium was aspirated, washed 3 times with phosphate-balanced physiological saline (PBS), and cleared of Al-1 and AL @ M-1 that were not ingested by phagocytosis;
6) PBS was blotted dry and 0.5mL of paraformaldehyde (4%) was added to each well for fixation at room temperature for 10 min;
7) washing with PBS for 3 times, and removing residual paraformaldehyde fixing solution;
8) adding 150 microliters of fluorescent dye Hochest3342 into each hole for dyeing, and coloring cell nuclei;
9) after 3 washes with PBS, 200 microliters of anti-fluorescence quencher was added to each well;
10) fluorescence microscope photographing observation (excitation light: hochest3342-340 nm, FITC-525 nm).
Fluorescence microscopy results As shown in FIG. 1, both AL-1 and AL @ M-1 exhibited green fluorescence by coating with FITC (green fluorescence) which represents the intracellular uptake and distribution of AL-1 and AL @ M-1, respectively. As shown in FIG. 1, cardiomyocytes phagocytose AL @ M-1 more than AL-1. And the uptake of AL-1 and AL @ M-1 in the myocardial cells was further quantified by flow cytometry. The results show that the mean immunofluorescence intensity of myocardial cell phagocytosis AL-1 is 4430 + -219; phagocytosis AL @ M-1 mean immunofluorescence intensity was 14633+463, with a significant increase in phagocytic efficiency (p < 0.001).
Example 2:
mixing a microRNA (anti-miR-133 antisense oligonucleotide, AMO-133 for short) solution with the concentration of 1.5 wt% with RDS type hectorite (RDS-Lp) according to the liquid-solid ratio of 40:1, performing exchange adsorption for 10.0h at 50 ℃, centrifuging to obtain a solid product, washing for multiple times, and centrifuging to obtain the slow-release [ AMO-133/RDS-Lp ] -2 composite delivery material (AL-2 for short). Adding the obtained composite material into a Macrophage Membrane (MM) aqueous solution (liquid-solid ratio 40:1) with the concentration of 1.5 wt%, stirring, adsorbing and wrapping for 12.0h at 25 ℃, performing centrifugal separation, and preparing into a solution with the concentration of 1.5 wt%, thus obtaining the hectorite composite delivery material [ AMO-133/RDS-Lp @ MM ] -1 (AL @ M-2 for short) for targeted treatment of myocardial infarction.
The flow cytometry Annexin V/PI double staining method is used for evaluating the influence of AL @ M-2 on myocardial cell apoptosis under an oxidative stress state:
1) cell collection: directly collecting the suspension cells into a 10ml centrifuge tube, wherein the number of the cells in each sample is (1-5) multiplied by 106Centrifuging for 5min at 500-1000 r/min per mL, and discarding the culture solution.
2) Washing with incubation buffer solution for 1 time, and centrifuging for 5min at 500-1000 r/min.
3) Resuspend the cells with 100ul of labeling solution, incubate in dark at room temperature for 10-15 min.
4) Centrifuging at 500-1000 r/min for 5min, washing the settled cells with the incubation buffer for 1 time.
5) Incubate 20min with addition of fluorescent (SA-FLOUS) solution 4, protected from light and shaking occasionally.
6) Flow cytometry analysis: the wavelength of the excitation light of the flow cytometer is 488nm, the fluorescence of FITC is detected by a band-pass filter with the wavelength of 515nm, and PI is detected by another filter with the wavelength of more than 560 nm.
Flow cytometry results are shown in FIG. 2, comparing cardiomyocytes (a in FIG. 2), H in normal environment2O2Apoptosis of myocardial cells was significantly up-regulated after stimulation (b in FIG. 2), and H was significantly alleviated after the addition of laponite nanocomposite AL @ M-22O2Degree of apoptosis of cardiac cells after stimulation (c in fig. 2).
Example 3:
mixing a microRNA (anti-miR-92 a antisense oligonucleotide, AMO-92a for short) solution with the concentration of 2.0 wt% and S482 type hectorite (S482-Lp) according to the liquid-solid ratio of 40:1, performing exchange adsorption for 10.0h at 50 ℃, centrifuging to obtain a solid product, washing for many times, and centrifuging to obtain the sustained-release [ AMO-92a/S482] -3 composite delivery material (AL-3 for short). Adding the obtained composite material into a platelet exosome (PLT-EV) aqueous solution (liquid-solid ratio 40:1) with the concentration of 2.0 wt%, stirring, adsorbing and wrapping for 12.0h at 25 ℃, performing centrifugal separation, and preparing into a solution with the concentration of 2.0 wt%, thus obtaining the hectorite composite delivery material [ AMO-92a/S482-Lp @ PLT-EV ] -3 (AL @ M-3 for short) for targeted treatment of myocardial infarction.
The targeting of AL-3 and AL @ M-3 to the heart and the distribution of other organs are observed through in vitro activity imaging:
1) the FITC label AL-3 and AL @ M-3 were prepared in advance: 100mg AL-3 and 100mg AL @ M-3 solutions are respectively dispersed in 8mL deionized water, 3mL FITC ethanol solution (1mL/mg) is added, after stirring for 6h in the dark, 4000g of nanoparticles are collected by centrifugation, washed for 3 times, filtered by a 0.45 mu M filter membrane, and then sampled and quantified for later use. Each group of rats was given 1mg/kg doses of AL-3 and AL @ M-3 injections 6 hours prior to testing.
2) Anesthesia: each rat was weighed and given 2% pentobarbital sodium solution i.p. 300 μ l/100 g;
3) after the rat is completely anesthetized, the skin is incised at the 3-4 intercostal transverse incision at the left side of the sternum, hemostatic forceps are used for separating each muscle layer by layer, after the pleura is punctured, a rib spreader is used for spreading the ribs, the pericardium is exposed, the pericardium is carefully torn, and the heart is exposed;
4) carefully cutting off the heart, putting the heart in precooled PBS (phosphate buffer solution) to wash out surface bloodstains, and sucking the surface moisture;
5) placing the heart on black background test paper, sending the heart into a small animal living body optical two-dimensional imaging system to detect a fluorescence signal FITC: 488 nm;
6) data were recorded and image analyzed.
MI/RI rats were imaged ex vivo with heart by using FITC labeled AL-3 and AL @ M-3. After intravenous injection of FITC-labeled AL-3 and AL @ M-36 hours in MI/RI rats, uptake in injured tissue was increased by approximately 5-fold in the AL @ M-3 group as quantified by radiation efficiency (FIG. 3).
Example 4:
mixing a microRNA (anti-miR-204 antisense oligonucleotide, AMO-204 for short) solution with the concentration of 1.5 wt% with XLS type hectorite (XLS-Lp) according to the liquid-solid ratio of 40:1, performing exchange adsorption for 8.0h at 50 ℃, centrifuging to obtain a solid product, washing for multiple times, and centrifuging to obtain the slow-release [ AMO-204/XLS-Lp ] -4 composite delivery material (AL-4 for short). Adding the obtained composite material into a platelet exosome (PLT-EV) aqueous solution (liquid-solid ratio 40:1) with the concentration of 1.5 wt%, stirring, adsorbing and wrapping for 10.0h at 25 ℃, performing centrifugal separation, and preparing into a solution with the concentration of 1.5 wt%, thus obtaining the hectorite composite delivery material [ AMO-204/XLS-Lp @ PLT-EV ] -4 (AL @ M-4 for short) for targeted treatment of myocardial infarction.
Through the fluorescent microscope positioning and in vitro activity imaging detection and observation, the phagocytic uptake efficiency of the myocardial cells to the AL @ M-4 and the targeting capability of the AL @ M-4 to the injured myocardium in vivo show better characteristics.
Example 5:
mixing a microRNA (anti-miR-133 antisense oligonucleotide, AMO-133 for short) solution with the concentration of 1.5 wt% with RDS type hectorite (RDS-Lp) according to the liquid-solid ratio of 60:1, performing exchange adsorption for 10.0h at 50 ℃, centrifuging to obtain a solid product, washing for multiple times, and centrifuging to obtain the slow-release [ AMO-133/RDS-Lp ] -5 composite delivery material (AL-5 for short). Adding the obtained composite material into a Macrophage Membrane (MM) aqueous solution (liquid-solid ratio 60:1) with the concentration of 1.5 wt%, stirring, adsorbing and wrapping for 12.0h at 25 ℃, performing centrifugal separation, and preparing into a solution with the concentration of 1.5 wt%, thus obtaining the hectorite composite delivery material [ AMO-133/RDS-Lp @ MM ] -5 (AL @ M-5 for short) for targeted treatment of myocardial infarction.
Through the fluorescent microscope positioning and the immune flow type apoptosis detection observation, the phagocytic uptake efficiency of the myocardial cells to AL @ M-5 and AL @ M-5 to H2O2The anti-apoptosis ability of the myocardial cells after stimulation shows better characteristics.
Example 6:
mixing a microRNA (anti-miR-1 antisense oligonucleotide, AMO-1 for short) solution with the concentration of 2.0 wt% with S482 type hectorite (S482-Lp) according to the liquid-solid ratio of 40:1, performing exchange adsorption at 50 ℃ for 10.0h, centrifuging to obtain a solid product, washing for multiple times, and centrifuging to obtain the sustained-release [ AMO-1/S482] -6 composite delivery material (AL-6 for short). Adding the obtained composite material into a platelet exosome (PLT-EV) aqueous solution (liquid-solid ratio 40:1) with the concentration of 2.0 wt%, stirring, adsorbing and wrapping for 12.0h at 25 ℃, performing centrifugal separation, and preparing into a solution with the concentration of 2.0 wt%, thus obtaining the hectorite composite delivery material [ AMO-1/S482-Lp @ PLT-EV ] -6 (AL @ M-6 for short) for targeted treatment of myocardial infarction.
Flow cytometric Annexin V/PI double staining method is used for evaluating the influence of AL @ M-6 on myocardial apoptosis under oxidative stress state, and the result shows that AL @ M-6 is applied to H2O2The anti-apoptosis ability of the myocardial cells after stimulation shows better characteristics; and shows excellent cell phagocytosis and tissue targeting in fluorescence microscope and in vitro activity imaging detection.
The embodiments described above are intended to facilitate a person of ordinary skill in the art in understanding and using the invention. It will be readily apparent to those skilled in the art that various modifications to these embodiments may be made, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above embodiments, and those skilled in the art should make improvements and modifications within the scope of the present invention based on the disclosure of the present invention.
Claims (10)
1. The hectorite composite delivery material for targeted therapy of myocardial infarction is characterized by comprising a hectorite loaded with microRNA and an externally coated targeted specific cell membrane.
2. The laponite complex delivery material for targeted therapy of myocardial infarction of claim 1, wherein the laponite comprises one or more of RD laponite, RDs laponite, XLG laponite, XLS laponite, or S482 laponite, the microRNA comprises one or more of miR-1, miR-204, miR-133a, or miR-92a, and the target specific cell membrane is one of platelet membrane, macrophage membrane, or exosome.
3. The hectorite composite delivery material for targeted therapy of myocardial infarction according to claim 1, wherein the mass percentage of microRNA in the composite delivery material is 0.01-20%, and the mass percentage of the targeted specific cell membrane is 0.01-20%.
4. The hectorite composite delivery material for targeted therapy of myocardial infarction according to claim 1, wherein the composite delivery material further comprises a solvent, and the mass ratio of the solvent to the hectorite is 4-1000: 1.
5. The method for preparing the hectorite composite delivery material for targeted therapy of myocardial infarction according to any one of claims 1 to 4, wherein the method comprises the following steps:
1) mixing the microRNA solution with hectorite, carrying out exchange adsorption reaction, and separating to obtain a slow-release microRNA-hectorite composite material;
2) dispersing the slow-release microRNA-hectorite composite material in a solvent, adding a target specific cell membrane, carrying out adsorption and coating reaction under stirring, and separating to obtain the hectorite composite delivery material for targeted treatment of myocardial infarction.
6. The preparation method of the hectorite composite delivery material for targeted therapy of myocardial infarction according to claim 5, wherein in the step 1), the concentration of the microRNA solution is 0.5 wt% -5 wt%, the temperature of the exchange adsorption reaction is 0-50 ℃, and the time is 0.1-20 h.
7. The preparation method of the laponite composite delivery material for targeted therapy of myocardial infarction according to claim 5, wherein in the step 2), the solvent is water, and the temperature of the adsorption and coating reaction is 0-50 ℃ and the time is 1-48 h.
8. The method for preparing the hectorite composite delivery material for targeted therapy of myocardial infarction according to claim 5, wherein in the step 2), the product obtained by separation after the adsorption and coating reaction is added into a solvent to prepare a solution.
9. Use of the composite delivery material according to any one of claims 1 to 4 for the preparation of a medicament for the targeted treatment of myocardial infarction.
10. A medicament for targeted therapy of myocardial infarction, characterized in that it comprises a composite delivery material according to any one of claims 1 to 4.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060069055A1 (en) * | 2004-09-21 | 2006-03-30 | Maya Dajee | Delivery of polynucleotides |
| WO2014057203A1 (en) * | 2012-10-09 | 2014-04-17 | Societe D'exploitation De Produits Pour Les Industries Chimiques Seppic | Pharmaceutical compositions comprising capsules obtained by coacervation without the use of toxic cross-linking agents |
| CN105189784A (en) * | 2013-05-07 | 2015-12-23 | 精密公司 | Device for preparation and analysis of nucleic acids |
| US20180021253A1 (en) * | 2015-01-30 | 2018-01-25 | President And Fellows Of Harvard College | Peritumoral and intratumoral materials for cancer therapy |
| EP3285782A1 (en) * | 2015-04-22 | 2018-02-28 | Innolife Co., Ltd. | Methods of tissue repair and regeneration |
-
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- 2022-01-12 CN CN202210033642.6A patent/CN114469897B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060069055A1 (en) * | 2004-09-21 | 2006-03-30 | Maya Dajee | Delivery of polynucleotides |
| WO2014057203A1 (en) * | 2012-10-09 | 2014-04-17 | Societe D'exploitation De Produits Pour Les Industries Chimiques Seppic | Pharmaceutical compositions comprising capsules obtained by coacervation without the use of toxic cross-linking agents |
| CN105189784A (en) * | 2013-05-07 | 2015-12-23 | 精密公司 | Device for preparation and analysis of nucleic acids |
| US20160090588A1 (en) * | 2013-05-07 | 2016-03-31 | Micronics, Inc. | Device for preparation and analysis of nucleic acids |
| US20180021253A1 (en) * | 2015-01-30 | 2018-01-25 | President And Fellows Of Harvard College | Peritumoral and intratumoral materials for cancer therapy |
| EP3285782A1 (en) * | 2015-04-22 | 2018-02-28 | Innolife Co., Ltd. | Methods of tissue repair and regeneration |
| US20180099008A1 (en) * | 2015-04-22 | 2018-04-12 | Innolife Co., Ltd. | Methods of tissue repair and regeneration |
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