CN112618487A - Thrombus-targeted long-circulating polycation micelle and preparation method and application thereof - Google Patents
Thrombus-targeted long-circulating polycation micelle and preparation method and application thereof Download PDFInfo
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- CN112618487A CN112618487A CN202011254555.0A CN202011254555A CN112618487A CN 112618487 A CN112618487 A CN 112618487A CN 202011254555 A CN202011254555 A CN 202011254555A CN 112618487 A CN112618487 A CN 112618487A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
- A61K9/1075—Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
- A61K31/727—Heparin; Heparan
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/55—Protease inhibitors
- A61K38/57—Protease inhibitors from animals; from humans
- A61K38/58—Protease inhibitors from animals; from humans from leeches, e.g. hirudin, eglin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—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
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—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
- 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
- A61K47/51—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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
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Abstract
The invention discloses a thrombus-targeted long-circulating polycation micelle and a preparation method and application thereof. The thrombus-targeted polyion micelle prepared by the invention can achieve the purposes of enhancing the stability of protein drugs, targeting thrombus, prolonging the action time in vivo and reducing the immunogenicity. The preparation method is feasible and reliable in operation, the obtained carrier has high drug loading, the drug has obvious thrombus targeting property, the in-vivo action time is prolonged, and a theoretical basis is provided for the research of a novel thrombus targeting drug delivery system.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a thrombus-targeted long-circulating polycation micelle, and a preparation method and application thereof.
Background
Cardiovascular and cerebrovascular diseases are the first fatal diseases at present, and seriously threaten the life health of human beings. Ischemic stroke, myocardial infarction and venous thromboembolism caused by thrombus are three killers of cardiovascular and cerebrovascular diseases. The drug thrombolysis is a common method for treating thrombus clinically. However, the anticoagulant and antithrombotic drugs lack tissue specificity, cannot selectively act on the diseased region, and are easy to cause serious complications such as bleeding, and the clinical application of the drugs is greatly limited due to the defects. Therefore, the research on a novel drug delivery system of an anticoagulant and antithrombotic drug has attracted the attention of a plurality of scholars, and in recent years, novel preparations such as subcutaneous injection temperature-sensitive gel, polyethylene glycol preparation, nasal spray and the like are developed. Although the preparations can prolong the in vivo circulation time of the medicine, the targeting is insufficient, the absolute bioavailability is still low, and a large amount of organic solvents are used in the preparation process, so that the toxicity to the organism is easily caused. Therefore, the development of an effective preparation which can carry and deliver the anticoagulant and antithrombotic drug to the thrombus in a targeted manner, can prolong the circulation time of the drug in vivo and has low toxicity is urgently needed, the specific targeting property of the anticoagulant and antithrombotic drug is enhanced, and the prolonging of the action time of the drug in vivo is a key measure for reducing the dosage, improving the bioavailability and reducing the side effect.
Currently, enhanced drug targeting can be achieved through specific recognition of ligand-receptors. Arginine-glycine-aspartic acid sequence (Arg-Gly-Asp, RGD) short peptides are recognition sites for the binding of cell membrane integrin receptors to IIb/IIIa globulin (GP IIb/IIIa) or other extracellular ligands on activated platelets. On resting platelets, GP IIb/IIIa is in an inactive state and cannot bind to the RGD fragment of adhesion proteins. However, once the endothelium is damaged and the platelets adhere to the endothelium, after an inducer (such as thrombin, collagen, thrombin-sensitive protein, ADP, thromboxane a2, and the like) is combined with a corresponding receptor, the steric configuration activation state of GP IIb/IIIa is regulated through different intracellular signal transduction mechanisms to be activated, the adhesion capability of GP IIb/IIIa and fibrinogen is improved, and the adhesion and aggregation of the platelets are accelerated. The RGD tripeptide is used as a target molecule for guiding and activating the platelet, can bring anticoagulant and antithrombotic drugs to the vicinity of thrombus, competitively combines and activates GP IIb/IIIa receptors on the platelet, antagonizes the combination of the platelet and fibrin, and reduces the adhesion and aggregation of the platelet. Therefore, the RGD peptide can be used as a guiding warhead of a thrombus targeting preparation. The RGD gene is recombined to the gene of hirudin, and the hirudin with anticoagulation and antiplatelet functions is obtained by fermentation, but the method has high production cost, changes the original chemical composition of the medicament, and the structural modification of the hirudin can potentially influence the curative effect and the elimination of the medicament. Therefore, the development of a novel preparation which does not change the chemical composition of the original medicine, has simple preparation process and long in-vivo circulation time and is targeted on thrombus is still urgent.
In recent years, polyion complex micelles (PIC micelles) have attracted great attention with their unique properties. PIC micelles are typically spontaneously assembled to form micelles with a core-shell structure by electrostatic interactions between a charged polymer and an oppositely charged species as the driving force for the formation of the hydrophobic core of the micelle. The PIC micelle has the characteristics of the traditional polymer micelle: stable structure, targeting property, long circulation time in vivo, good safety, simple preparation and the like. In addition, PIC micelles have their unique advantages: the drug loading range is wide; the drug loading process is basically carried out in the aqueous solution, so that the use of organic solvent is avoided, and the toxic and side effects caused by solvent residue can be eliminated.
Currently, cationic polymers used for preparing PIC micelles include poly-L-lysine, polydimethylaminoethyl methacrylate, polyethyleneimine, chitosan, and the like. Among these polymers, chitosan is the only basic polysaccharide in nature, chemically named (1,4) -2-amino-2-deoxy- β -D-glucan, a cationic polymer with biocompatibility, biodegradability and pH sensitivity. In a weakly acidic medium, amino groups in chitosan molecules are protonated to enable chitosan to be positively charged, and the chitosan can be aggregated with certain negatively charged substances (such as electronegative drugs) through electrostatic interaction, so that the chitosan can be used as a polycation chain segment of a PIC micelle. Poly (2-ethyl-2-oxazoline) (PEOz) is selected as a hydrophilic fragment of the PIC micelle, the PEOz has good biocompatibility, passes the certification of the U.S. food and drug administration, can be used for modifying the surface of the micelle, can increase the surface hydrophilicity and the biocompatibility, and simultaneously, PEOz chains can form steric hindrance so that the micelle is hidden from the recognition of a reticuloendothelial system and is prevented from being absorbed and removed, thereby prolonging the internal circulation time of the medicament. Meanwhile, PEOz has lower immunogenicity compared with PEG, and can improve the defects that PEG preparations induce strong immune response when repeatedly used and cause the blood circulation time of the preparations to be shortened (ABC effect).
Therefore, the RGD peptide modified PEOz-chitosan polyion micelle is expected to realize the goals of anticoagulation and thrombus prevention of thrombus targeting and in vivo long circulation, and no report is found on the RGD-PEOz-chitosan polycation micelle and the application thereof in thrombus targeting treatment at present.
Disclosure of Invention
The invention aims to provide a thrombus-targeted long-circulating polycation micelle which can realize the aims of thrombus targeting, long circulation and low immunogenicity after an antithrombotic drug is loaded.
The second object of the present invention is to provide a method for preparing the above micelle.
It is a third object of the present invention to provide the use of the above micelle.
The first technical scheme adopted by the invention is as follows: a thrombus-targeted long-circulating polycation micelle is prepared by preparing polyion micelle by poly (2-ethyl-2-oxazoline) grafted chitosan through an EDC/NSH esterification system, modifying arginine-glycine-aspartic acid sequence short peptide at the chain end, and loading an anticoagulant and antithrombotic drug.
The second technical scheme adopted by the invention is as follows: a preparation method of a thrombus-targeted long-circulating polycation micelle specifically comprises the following steps:
step 2, dissolving the HOOC-PEOZ-OH powder obtained in the step 1 in dimethyl sulfoxide, adding acetic anhydride, magnetically stirring under the protection of nitrogen, after the reaction is finished, rotationally evaporating to remove the organic solvent in the reaction liquid, dropwise adding a small amount of dichloromethane for redissolution, precipitating with excessive cold ethyl ether, and performing suction filtration to obtain HOOC-PEOz-CHO solid powder;
step 3, dissolving chitosan in distilled water, and stirring until the chitosan is completely dissolved to prepare a solution I; dissolving the HOOC-PEOz-CHO solid powder prepared in the step 2 in absolute ethyl alcohol, dissolving NHS and EDC in absolute ethyl alcohol, pouring the absolute ethyl alcohol mixed solution containing HOOC-PEOz-CHO into the dissolved absolute ethyl alcohol mixed solution of NHS and EDC, stirring and dissolving at 65 ℃ to prepare a solution II; adding the solution II into the solution I under vigorous stirring, reacting at 65 ℃, putting the solution into a dialysis bag after reaction, dialyzing in distilled water, freeze-drying, washing the obtained product with absolute ethyl alcohol, and performing suction filtration to obtain CHO-PEOz-CS;
step 4, stirring and dissolving the CHO-PEOz-CS obtained in the step 3 and RGD in 95% ethanol with the pH of 4.6, and then dissolving NaBH3CN and CHO-PEOz-CS are added into the mixture in a certain molar ratioDialyzing the obtained product in distilled water, and freeze-drying to obtain a product RGD-PEOz-CS;
and 5, precisely weighing RGD-PEOz-CS, dissolving the RGD-PEOz-CS in 1 acetate buffer solution, adding an equivalent volume of an antithrombotic drug solution, wherein the mass ratio of the antithrombotic drug to the RGD-PEOz-CS is 1: 2-10, adding a TPP solution with the concentration of 1-2 mg/mL into the solution under room-temperature magnetic stirring, and continuously stirring for 30-90 min after adding to obtain the drug-loaded RGD-PIC polyion micelle.
The second technical solution adopted by the present invention is further characterized in that,
in the step 1, the mass ratio of the monomer EOz to the 3-bromo-ethyl propionate is 1-5: 1, the volume of acetonitrile is 80-250 mL, the molar ratio of the 3-bromo-ethyl propionate to the potassium iodide is 1: 0.5-2, the stirring speed is 300-800 rpm, the first heating reflux reaction time is 24-48 h, the temperature is 80-120 ℃, the molar ratio of potassium hydroxide to the 3-bromo-ethyl propionate is 2-5: 1, the second heating reflux reaction time is 12-24 h, the temperature is 80-120 ℃, the rotary evaporation temperature is 40-80 ℃, the volume of dichloromethane is 10-20 mL, the volume of cold ether is 200-500 mL, the volume of water is 10-20 mL, the cut-off molecular weight of a dialysis bag is 8000-14000, the dialysis time is 2-4 days, and freeze drying is 1-2 days.
HOOC-PEO in step 2ZThe mass of the-OH powder is 0.5-2 g, the volume of the dimethyl sulfoxide is 2-10 mL, the volume of the acetic anhydride is 0.1-0.5 mL, the stirring speed is 300-800 rpm, the stirring time is 8-24 h, the rotary evaporation temperature is 40-60 ℃, the volume of the dichloromethane is 10-20 mL, the volume of the cold ether is 200-500 mL, and the dialysis time is 2-4 days.
In the step 3, the mass of the chitosan is 0.2-1 g, the volume of the distilled water is 30-150 mL, the first stirring speed is 300-800 rpm, the mass of HOOC-PEOz-CHO is 0.1-0.5 g, the volume of absolute ethyl alcohol for dissolving HOOC-PEOz-CHO is 20-100 mL, the mass of NHS is 0.1-0.5 g, the mass of EDC is 0.1-0.5 g, the volume of absolute ethyl alcohol for dissolving NHS and EDC is 50-250 mL, the second stirring speed is 300-800 rpm, the reaction time is 3-6 h, the cut-off molecular weight of a dialysis bag is 8000-14000, the dialysis time is 2-4 days, and the freeze drying is 1-2 days.
In the step 4, the molar ratio of CHO-PEOz-CS to RGD is 1: 0.2-0.6, and the CHO-PEOz-CS and NaBH are3CN is 1: 0.2-0.6, the cut-off molecular weight of the dialysis bag is 8000-14000, the dialysis time is 2-4 days, and the freeze drying is 1-2 days.
The antithrombotic drug in the step 5 is any one of heparin, natural hirudin, recombinant hirudin, streptokinase or urokinase.
In the step 5, the mass of the RGD-PEOz-CS is 10-20 mg, the volume of the acetate buffer solution is 10-20 mL, the stirring speed is 300-600 rpm, and the volume of the TPP solution is 8-10 mL.
The third technical scheme adopted by the invention is as follows: an application of a thrombus-targeted polyion micelle in the field of antithrombotic drug delivery.
The invention has the beneficial effects that: the thrombus-targeted polyion micelle prepared by the invention can achieve the purposes of enhancing the stability of protein drugs, targeting thrombus, prolonging the action time in vivo and reducing the immunogenicity. The preparation method is feasible and reliable in operation, the obtained carrier has high drug loading, the drug has obvious thrombus targeting property, the in-vivo action time is prolonged, and a theoretical basis is provided for the research of a novel thrombus targeting drug delivery system.
Drawings
FIG. 1 is an infrared spectrum of RGD-PEOz-CS according to the present invention;
fig. 2 is a particle size distribution diagram of the drug-loaded micelle of the present invention.
Detailed Description
The invention is further described with reference to the following figures and detailed description.
A preparation method of a thrombus-targeted long-circulating polycation micelle specifically comprises the following steps:
And 2, weighing 0.5-2 g of HOOC-PEOZ-OH powder obtained in the step 1, dissolving the powder in 2-10 mL of dimethyl sulfoxide, adding 0.1-0.5 mL of acetic anhydride, magnetically stirring for 8-24 h under the protection of nitrogen, removing the organic solvent in the reaction solution by rotary evaporation at the stirring speed of 300-800 rpm at 40-60 ℃, dropwise adding 10-20 m of dichloromethane for redissolution, precipitating with 200-500 mL of cold ether, and performing suction filtration to obtain the HOOC-PEOz-CHO solid powder.
Step 3, weighing 0.2-1 g of chitosan, dissolving the chitosan in 30-150 mL of distilled water, and stirring at the rotating speed of 300-800 rpm until the chitosan is completely dissolved to prepare a solution I; dissolving the HOOC-PEOz-CHO obtained in the step 2 in 20-100 mL of absolute ethyl alcohol, dissolving 0.1-0.5 g of NHS and 0.1-0.5 g of EDC in 50-250 mL of absolute ethyl alcohol, pouring the absolute ethyl alcohol mixed solution containing the HOOC-PEOz-CHO into the dissolved absolute ethyl alcohol mixed solution of the NHS and the EDC, and stirring and dissolving at 65 ℃ to prepare a solution II;
and stirring the solution II at the rotation speed of 300-800 rpm, adding the solution II into the solution I, reacting for 3-6 h at 65 ℃, placing the solution after reaction into a dialysis bag with the molecular weight cutoff of 8000-14000, dialyzing for 2-4 days in distilled water, freeze-drying for 1-2 days, washing the obtained product with absolute ethyl alcohol, and performing suction filtration to obtain the CHO-PEOz-CS.
Step 4, stirring and dissolving the CHO-PEOz-CS and the RGD obtained in the step 3 in 95% ethanol with the pH of 4.6, wherein the molar ratio of the CHO-PEOz-CS to the RGD is 1: 0.2-0.6, and then adding NaBH3Adding CN and CHO-PEOz-CS into the mixture in a molar ratio of 1: 0.2-0.6, and placing the mixture into a cut-offDialyzing in distilled water for 2-4 days in a dialysis bag with the molecular weight of 8000-14000, and freeze-drying the dialysis bag for 1-2 days to obtain a product RGD-PEOz-CS;
accurately weighing 10-20 mg of RGD-PEOz-CS, dissolving in 10-20 mL of acetate buffer solution, adding an equivalent volume of an antithrombotic drug solution, wherein the mass ratio of the antithrombotic drug to the RGD-PEOz-CS is 1: 2-10, adding 8-10 mL of TPP solution with the concentration of 1-2 mg/mL into the solution under room-temperature magnetic stirring (300-600 rpm), and continuing to stir for 30-90 min after adding to obtain the drug-loaded RGD-PIC polyion micelle.
The antithrombotic is any one of heparin, natural hirudin, recombinant hirudin, streptokinase or urokinase.
Compared with the prior art, the invention has the following advantages:
(1) the RGD is modified on the surface of the polyion micelle, and can be combined with GP IIb/IIIa receptors on activated platelets to realize thrombus targeting.
(2) The invention takes PEOz as the hydrophilic chain segment of the polyion micelle, can perform hydrophilic modification, and prolongs the in vivo circulation time of the medicament;
(3) the invention takes PEOz as the hydrophilic chain segment of the polyion micelle, can improve the defect that the PEG preparation induces strong immune response when repeatedly used, which leads to the shortening of the blood circulation time (ABC effect) of the preparation;
(4) the preparation method has the advantages of simple operation, easy obtaining of products, simple post-treatment and suitability for industrial production.
Example 1
Placing 10g of anhydrous EOz into a round-bottom flask, adding 80mL of anhydrous acetonitrile, adding 3-bromo-ethyl propionate and potassium iodide (the molar ratio is 1:0.5) according to the mass ratio of a monomer to an initiator being 1:1, stirring (300rpm) under the protection of nitrogen, heating and refluxing at 80 ℃ for 24 hours, adding a potassium hydroxide methanol solution to stop the reaction after the reaction liquid is cooled to room temperature (the molar ratio of potassium hydroxide to 3-bromo-ethyl propionate being 1:1), heating and refluxing at 80 ℃ for 12 hours, removing an organic solvent in the reaction liquid by rotary evaporation at 40 ℃, redissolving with 10mL of dichloromethane, precipitating with 200mL of cold ether, and carrying out suction filtration. Dissolving the obtained solid in 10mL of water, placing in a dialysis bag with molecular weight of 8000, dialyzing in deionized water for 24h, and freeze-drying for 1 day to obtain HOOC-PEOZ-OH powder.
Step 2, HOOC-PEOZSynthesis of-CHO
0.5g of HOOC-PEOZ-OH was dissolved in 2mL of dimethyl sulfoxide, 0.1mL of acetic anhydride was added, and the mixture was magnetically stirred under nitrogen for 8h at a stirring speed of 300 rpm. After the reaction is finished, the organic solvent in the reaction solution is removed by rotary evaporation at 40 ℃, 10mL of dichloromethane is dripped for redissolution, 200mL of cold ether is used for precipitation and suction filtration, and then HOOC-PEOz-CHO solid powder is obtained.
Step 3, synthesis of CHO-PEOz-CS
Dissolving 0.2g of chitosan in 30mL of distilled water, and stirring (300rpm) until the chitosan is completely dissolved to prepare a solution I; dissolving 0.1g of HOOC-PEOz-CHO in 20mL of absolute ethyl alcohol, dissolving 0.1g of NHS and 0.1g of EDC in 50mL of absolute ethyl alcohol, pouring the absolute ethyl alcohol mixed solution containing the HOOC-PEOz-CHO into the dissolved anhydrous ethyl alcohol mixed solution of the NHS and the EDC, and stirring and dissolving at 65 ℃ to prepare a solution II;
adding the solution II into the solution I under the condition of vigorous stirring (600rpm), reacting at 65 ℃, putting the reacted solution into a dialysis bag (molecular weight cut-off 8000) to dialyze in distilled water for 2d, and freeze-drying for 1d to obtain the CHO-PEOz-CS.
Step 4, synthesis of RGD-PEOz-CS
CHO-PEOz-CS and RGD are dissolved in 95% ethanol with pH 4.6 at a molar ratio of 1:0.2 with stirring for 24h, and then NaBH is added3CN and CHO-PEOz-CS are added into the mixture in a molar ratio of 1:0.2, the mixture is dialyzed in distilled water for 2d by a dialysis bag with molecular weight cut-off of 8000, and the product RGD-PEOz-CS is obtained after freeze drying for 1 d;
Precisely weighing 10mg RGD-PEOz-CS, dissolving in 10mL of acetate buffer solution, adding an isometric drug solution (the mass ratio of the drug to the RGD-PEOz-CS is 1:2), adding 8mL of TPP solution (1mg/mL) into the solution under room temperature magnetic stirring (the rotating speed is 300rpm), and after the addition, continuing stirring for 30min to obtain the drug-loaded RGD-PIC polyion micelle.
Example 2
Step 2, synthesis of HOOC-PEOZ-OH
Placing 15g of anhydrous EOz in a round bottom flask, adding 100mL of anhydrous acetonitrile, adding 3-bromo-ethyl propionate and potassium iodide (the molar ratio is 1:1) according to the mass ratio of a monomer to an initiator of 2:1, stirring under the protection of nitrogen (400rpm), heating and refluxing at 100 ℃ for 36h, cooling the reaction liquid to room temperature, adding a potassium hydroxide methanol solution to terminate the reaction (the molar ratio of potassium hydroxide to 3-bromo-ethyl propionate is 2:1), and heating and refluxing at 100 ℃ for 12 h. The organic solvent in the reaction solution was removed by rotary evaporation at 50 ℃ and redissolved with 12mL of dichloromethane, precipitated with 250mL of cold ether, and filtered under suction. Dissolving the obtained solid in 15mL of water, placing in a dialysis bag with molecular weight of 10000, dialyzing in deionized water for 36h, and freeze-drying for 1d to obtain HOOC-PEOZ-OH powder.
Step 2, HOOC-PEOZSynthesis of-CHO
1.0g of HOOC-PEOZ-OH was dissolved in 4mL of dimethyl sulfoxide, 0.2mL of acetic anhydride was added, and the mixture was magnetically stirred under nitrogen for 12h at a stirring speed of 500 rpm. After the reaction is finished, the organic solvent in the reaction solution is removed by rotary evaporation at 50 ℃, 12mL of dichloromethane is dripped for redissolution, 250mL of cold ether is used for precipitation and suction filtration, and then HOOC-PEOz-CHO solid powder is obtained.
Step 3, synthesis of CHO-PEOz-CS
Dissolving 0.4g of chitosan in 50mL of distilled water, and stirring (300rpm) until the chitosan is completely dissolved to prepare a solution I; dissolving 0.2g of HOOC-PEOz-CHO in 30mL of absolute ethyl alcohol, dissolving 0.2g of NHS and 0.2g of EDC in 100mL of absolute ethyl alcohol, pouring the absolute ethyl alcohol mixed solution containing the HOOC-PEOz-CHO into the dissolved anhydrous ethyl alcohol mixed solution of the NHS and the EDC, and stirring and dissolving at 65 ℃ to prepare a solution II;
adding the solution II into the solution I under the condition of vigorous stirring (700rpm), reacting at 65 ℃, putting the reacted solution into a dialysis bag (with cut-off molecular weight of 10000) to dialyze in distilled water for 3d, and freeze-drying for 1d to obtain the CHO-PEOz-CS.
Step 4, synthesis of RGD-PEOz-CS
Dissolving CHO-PEOz-CS and RGD in 95% ethanol with pH 4.6 at a molar ratio of 1:0.3 under stirring for 24h, adding NaBH3CN and CHO-PEOz-CS dropwise at a molar ratio of 1:0.3 into the mixture, dialyzing the mixture with a dialysis bag with molecular weight cut-off of 10000 in distilled water for 3d, and freeze-drying for 1d to obtain RGD-PEOz-CS;
Precisely weighing 10mg RGD-PEOz-CS, dissolving in 10mL of acetate buffer solution, adding an equal volume of drug solution (the mass ratio of the drug to the RGD-PEOz-CS is 1:4), adding 10mL of TPP solution (1.2mg/mL) into the solution under room temperature magnetic stirring (the rotating speed is 400rpm), and after adding, continuing to stir for 40min to obtain the drug-loaded RGD-PIC polyion micelle.
Example 3
Placing 20g of anhydrous EOz into a round bottom flask, adding 150mL of anhydrous acetonitrile, adding 3-bromo-ethyl propionate and potassium iodide (molar ratio is 1:1.5) according to the mass ratio of a monomer to an initiator of 3:1, stirring under the protection of nitrogen (500rpm), heating at 110 ℃ for reflux reaction for 36h, adding a potassium hydroxide methanol solution to terminate the reaction after the reaction liquid is cooled to room temperature (the molar ratio of potassium hydroxide to 3-bromo-ethyl propionate is 3:1), and heating at 110 ℃ for reflux for 16 h. The organic solvent in the reaction solution was removed by rotary evaporation at 60 ℃ and redissolved with 15mL of dichloromethane, precipitated with 300mL of cold ether, and filtered under suction. Dissolving the obtained solid in 18mL of water, placing in a dialysis bag with a molecular weight of 12000, dialyzing in deionized water for 48h, and freeze-drying for 2 days to obtain HOOC-PEOZ-OH powder.
Step 2, HOOC-PEOZSynthesis of-CHO
1.5g of HOOC-PEOZ-OH was dissolved in 6mL of dimethyl sulfoxide, 0.3mL of acetic anhydride was added, and the mixture was magnetically stirred under nitrogen for 16h at a stirring speed of 600 rpm. After the reaction is finished, the organic solvent in the reaction solution is removed by rotary evaporation at 60 ℃, 15mL of dichloromethane is dripped for redissolution, 300mL of cold ether is used for precipitation and suction filtration, and then HOOC-PEOz-CHO solid powder is obtained.
Step 3, synthesis of CHO-PEOz-CS
Dissolving 0.6g of chitosan in 80mL of distilled water, and stirring (400rpm) until the chitosan is completely dissolved to prepare a solution I; dissolving 0.3g of HOOC-PEOz-CHO in 30mL of absolute ethyl alcohol, dissolving 0.3g of NHS and 0.3g of EDC in 150mL of absolute ethyl alcohol, pouring the absolute ethyl alcohol mixed solution containing the HOOC-PEOz-CHO into the dissolved anhydrous ethyl alcohol mixed solution of the NHS and the EDC, and stirring and dissolving at 65 ℃ to prepare a solution II;
adding the solution II into the solution I under the condition of vigorous stirring (800rpm), reacting at 65 ℃, putting the reacted solution into a dialysis bag (with the molecular weight cut-off of 12000) to dialyze in distilled water for 4 days, and freeze-drying for 2 days to obtain the CHO-PEOz-CS.
Step 4, synthesis of RGD-PEOz-CS
Dissolving CHO-PEOz-CS and RGD in 95% ethanol at pH 4.6 at a molar ratio of 1:0.4 under stirring, adding NaBH3CN and CHO-PEOz-CS dropwise at a molar ratio of 1:0.4 into the mixture, dialyzing the mixture with a dialysis bag with a cut-off molecular weight of 12000 in distilled water for 4d, and freeze-drying for 2d to obtain a product RGD-PEOz-CS;
Precisely weighing 15mg RGD-PEOz-CS, dissolving in 15mL of acetate buffer solution, adding an isometric drug solution (the mass ratio of the drug to the RGD-PEOz-CS is 1:6), adding 9mL of TPP solution (1.5mg/mL) into the solution under room temperature magnetic stirring (the rotating speed is 500rpm), and after adding, continuously stirring for 50min to obtain the drug-loaded RGD-PIC polyion micelle.
Example 4
Putting 25g of anhydrous EOz into a round-bottom flask, adding 200mL of anhydrous acetonitrile, adding 3-bromo-ethyl propionate and potassium iodide (the molar ratio is 1:1.8) according to the mass ratio of a monomer to an initiator of 4:1, stirring under the protection of nitrogen (600rpm), heating at 120 ℃ for reflux reaction for 36 hours, adding a potassium hydroxide methanol solution to terminate the reaction after the reaction liquid is cooled to room temperature (the molar ratio of potassium hydroxide to 3-bromo-ethyl propionate is 4:1), and heating at 115 ℃ for reflux for 20 hours. The organic solvent in the reaction solution was removed by rotary evaporation at 70 ℃ and redissolved with 18mL of dichloromethane, precipitated with 400mL of cold ether, and filtered under suction. Dissolving the obtained solid in 20mL of water, placing in a dialysis bag with a molecular weight of 12000, dialyzing in deionized water for 48h, and freeze-drying for 2 days to obtain HOOC-PEOZ-OH powder.
Step 2, HOOC-PEOZSynthesis of-CHO
1.8g of HOOC-PEOZ-OH was dissolved in 8mL of dimethyl sulfoxide, 0.4mL of acetic anhydride was added, and the mixture was magnetically stirred under nitrogen for 20h at a stirring speed of 800 rpm. After the reaction is finished, the organic solvent in the reaction solution is removed by rotary evaporation at 70 ℃, 18mL of dichloromethane is dripped for redissolution, and then the HOOC-PEOz-CHO solid powder is obtained by precipitation and suction filtration of 400mL of cold ether.
Step 3, synthesis of CHO-PEOz-CS
Dissolving 0.8g of chitosan in 80mL of distilled water, and stirring (500rpm) until the chitosan is completely dissolved to prepare a solution I; dissolving 0.4g of HOOC-PEOz-CHO in 40mL of absolute ethyl alcohol, dissolving 0.4g of NHS and 0.4g of EDC in 200mL of absolute ethyl alcohol, pouring the absolute ethyl alcohol mixed solution containing the HOOC-PEOz-CHO into the dissolved anhydrous ethyl alcohol mixed solution of the NHS and the EDC, and stirring and dissolving at 65 ℃ to prepare a solution II;
adding the solution II into the solution I under the condition of vigorous stirring (800rpm), reacting at 65 ℃, putting the reacted solution into a dialysis bag (with the molecular weight cut-off of 12000) to dialyze in distilled water for 3d, and freeze-drying for 2d to obtain the CHO-PEOz-CS.
Step 4, synthesis of RGD-PEOz-CS
CHO-PEOz-CS and RGD are dissolved in 95% ethanol with pH 4.6 under stirring at a molar ratio of 1:0.5, and NaBH is then added3CN and CHO-PEOz-CS are added into the mixture in a molar ratio of 1:0.5, the mixture is dialyzed in distilled water for 3d by a dialysis bag with a cut-off molecular weight of 12000, and the mixture is freeze-dried for 1d to obtain a product RGD-PEOz-CS;
Precisely weighing 18mg RGD-PEOz-CS, dissolving in 18mL of acetate buffer solution, adding an equal volume of drug solution (the mass ratio of the drug to the RGD-PEOz-CS is 1:8), adding 10mL of TPP solution (1.8mg/mL) into the solution under room temperature magnetic stirring (the rotating speed is 500rpm), and after adding, continuing to stir for 60min to obtain the drug-loaded RGD-PIC polyion micelle.
Example 5
Placing 30g of anhydrous EOz in a round bottom flask, adding 250mL of anhydrous acetonitrile, adding 3-bromo-ethyl propionate and potassium iodide (molar ratio is 1:2) according to the mass ratio of a monomer to an initiator of 5:1, stirring under the protection of nitrogen (800rpm), heating at 120 ℃ for reflux reaction for 48 hours, adding a potassium hydroxide methanol solution to stop the reaction after the reaction liquid is cooled to room temperature (the molar ratio of potassium hydroxide to 3-bromo-ethyl propionate is 2:1), and heating at 120 ℃ for reflux for 24 hours. The organic solvent in the reaction solution was removed by rotary evaporation at 80 ℃ and redissolved with 20mL of dichloromethane, precipitated with 500mL of cold ether and filtered with suction. Dissolving the obtained solid in 20mL of water, placing in a dialysis bag with molecular weight of 14000, dialyzing in deionized water for 4 days, and freeze-drying for 2 days to obtain HOOC-PEOZ-OH powder.
Step 2, HOOC-PEOZSynthesis of-CHO
2g of HOOC-PEOZ-OH was dissolved in 10mL of dimethyl sulfoxide, 0.5mL of acetic anhydride was added, and the mixture was magnetically stirred under nitrogen for 24h at a stirring speed of 800 rpm. After the reaction is finished, the organic solvent in the reaction solution is removed by rotary evaporation at 60 ℃, 20mL of dichloromethane is dripped for redissolution, and 500mL of cold ether is used for precipitation and suction filtration to obtain HOOC-PEOz-CHO solid powder.
Step 3, synthesis of CHO-PEOz-CS
Dissolving 1g of chitosan in 150mL of distilled water, and stirring (800rpm) until the chitosan is completely dissolved to prepare a solution I; dissolving 0.5g of HOOC-PEOz-CHO in 100mL of absolute ethyl alcohol, dissolving 0.5g of NHS and 0.5g of EDC in 250mL of absolute ethyl alcohol, pouring the absolute ethyl alcohol mixed solution containing the HOOC-PEOz-CHO into the dissolved anhydrous ethyl alcohol mixed solution of the NHS and the EDC, and stirring and dissolving at 65 ℃ to prepare a solution II;
adding the solution II into the solution I under the condition of vigorous stirring (800rpm), reacting at 65 ℃, putting the reacted solution into a dialysis bag (molecular weight cut-off 14000) to dialyze in distilled water for 4 days, and freeze-drying for 2 days to obtain the CHO-PEOz-CS.
Step 4, synthesis of RGD-PEOz-CS
CHO-PEOz-CS and RGD are dissolved in 95% ethanol with pH 4.6 under stirring at a molar ratio of 1:0.6, and then NaBH is added3CN and CHO-PEOz-CS are added into the mixture in a molar ratio of 1:0.6, the mixture is dialyzed in distilled water for 4d by a dialysis bag with a cut-off molecular weight of 14000, and the mixture is freeze-dried for 2d to obtain a product RGD-PEOz-CS;
Precisely weighing 20mg RGD-PEOz-CS, dissolving in 20mL of acetate buffer solution, adding an isometric drug solution (the mass ratio of the drug to the RGD-PEOz-CS is 1:10), adding 10mL of TPP solution (2mg/mL) into the solution under room temperature magnetic stirring (the rotating speed is 600rpm), and after the addition, continuing stirring for 90min to obtain the drug-loaded RGD-PIC polyion micelle.
Detecting the structure of RGD-PEOz-CS: taking RGD-PEOz-CS powder, tabletting by using KBr, and carrying out structure detection by using FTIR. FIG. 1 is an infrared spectrum of RGD-PEOz-CS, which is 1636.05cm from FIG. 1-1The peak at (a) should be a characteristic peak of the secondary amide associated carbonyl (vc ═ o) to which PEOz is attached to CS; 3300-3060 cm-1A peak is formed, and is known as a characteristic peak of a secondary amide association state (v NH); at 1550-1510 cm-1There is a peak, which should be known as the characteristic peak of the secondary amide association state (. beta.NH), indicating successful attachment of PEOz to CS, 1050.68cm-1At a distance of 1087.35cm-1At a distance of 1156.04cm-1The peak at (A) should be a characteristic peak of secondary amine (. nu.C-N), and CHO-PEOz-CS is presumed to have attached RGD.
Detecting the particle size distribution of the drug-loaded polyion micelle: taking the drug-loaded polyion micelle, and measuring the particle size and the particle size distribution of the micelle by a dynamic light scattering method. FIG. 2 is a particle size distribution diagram of the drug-loaded polyion micelle, and it can be known from FIG. 2 that the average particle size of the micelle is 297.07nm, and the polydispersity index (PDI) is (0.160. + -. 0.080), which shows that the micelle prepared by the invention has uniform particle size and good stability.
Detecting in vivo pharmacokinetic parameters of the drug-loaded polyion micelle:
preparation of FITC-labeled hirudin:
liquid a: preparation 5.3mg of FITC was weighed and added to 500 μ L of carbonate buffer (pH 9, 100 mmol/L); b, liquid: 11mg of hirudin were weighed into 5mL of carbonate buffer (pH 9, 100 mmol/L); mixing the solution a and the solution b, reacting for 4h in the dark, dialyzing for 2 days (the dialysis bag is 3400Da, and the dialysis medium is 1000mL), and freeze-drying to obtain FITC-labeled hirudin;
hirudin solution: 50mg of FITC-labeled hirudin is weighed and prepared into 0.5mg/mL solution by using 100mL of normal saline, namely the hirudin solution.
According to the preparation process of the embodiment 1, 10mg of PIC and RGD-PIC are precisely weighed respectively, the PIC and RGD-PIC are respectively dissolved in 10mL of acetate buffer solution, 10mL of hirudin solution is added (the mass ratio of the drug to the RGD-PEOz-CS is 1:2), 8mL of TPP solution (1mg/mL) is added into the solution under room temperature magnetic stirring (the rotating speed is 300rpm), and after the addition, the stirring is continued for 30min, so that hirudin PIC micelle and hirudin RGD-PIC micelle are obtained.
Three groups of tests are set, namely a hirudin solution group, a hirudin PIC micelle group and a hirudin RGD-PIC micelle group, wherein each group is provided with three parallel rats, 9 rats are weighed and marked, 2mg/kg of hirudin is administrated by rat tail vein injection according to the weight, the administration time is recorded as 0 point, and the blood sampling time of each rat is 10min, 20min, 30min, 1h, 2h, 4h, 6h and 8 h. 0.5ml blood samples were collected from the orbital sinus each time, the collected whole blood was centrifuged at 3000r for 15min, and the supernatant was separated and stored at-20 ℃ until analysis. The FITC-hirudin concentration in the plasma was determined by means of a fluorescence spectrophotometer. The in vivo pharmacokinetic parameters are shown in table 1.
TABLE 1 in vivo pharmacokinetic parameters
As can be seen from Table 1, the micelle group significantly prolonged the half-life and increased the in vivo circulation time as compared with the hirudin solution group; according to the AUC value, the micelle group has improved bioavailability compared with the hirudin solution group, and the AUC value of the RGD-PIC micelle group is maximum; from the values of clearance CL, the micelle group decreased clearance compared to the hirudin solution group; the MRT value shows that the micelle group prolongs the average retention time and increases the in vivo circulation time compared with the hirudin solution group; in general, the PIC micelle group and the RGD-PIC micelle group achieve the expected effect, and the RGD-PIC micelle group has the most obvious effect of prolonging the in-vivo action time.
RGD gene is recombined to hirudin gene in the prior art, and hirudin with anticoagulation and anti-platelet functions is obtained by fermentation, but the method has extremely high production cost, changes the original chemical composition of the medicament, and the structural modification of the hirudin can potentially influence the curative effect and the elimination of the medicament. The invention does not change the chemical composition of the original medicine, has simple preparation process, simultaneously PEOz is used as the polycation chain segment of the PIC micelle, can increase the surface hydrophilicity and the biocompatibility of the PIC micelle, forms steric hindrance and avoids being absorbed and removed, and simultaneously, PEOz has lower immunogenicity compared with PEG and can further reduce the removal of PEOz in blood, thereby achieving the purpose of prolonging the internal circulation time of the medicine.
Claims (9)
1. The thrombus-targeted long-circulating polycation micelle is characterized in that poly (2-ethyl-2-oxazoline) grafted chitosan is used for preparing polyion micelles through an EDC/NSH esterification system, arginine-glycine-aspartic acid sequence short peptide is used for modifying at the chain end, and anticoagulant and antithrombotic drugs are loaded to obtain the thrombus-targeted long-circulating polycation micelle.
2. The preparation method of the thrombus-targeted long-circulating polycation micelle as claimed in claim 1, which is characterized by comprising the following steps:
step 1, placing anhydrous EOz into a round-bottom flask, adding anhydrous acetonitrile, adding a certain amount of 3-bromo-ethyl propionate and potassium iodide according to the molecular weight of a polymer to be synthesized and the proportion of a monomer and an initiator, stirring under the protection of nitrogen, heating and refluxing for reaction, adding a potassium hydroxide methanol solution to terminate the reaction after a reaction solution is cooled to room temperature, heating and refluxing, performing rotary evaporation to remove an organic solvent in the reaction solution, redissolving with dichloromethane, precipitating with excessive cold diethyl ether, performing suction filtration, dissolving a solid obtained by suction filtration in water, placing the solid in a dialysis bag, dialyzing in deionized water, and freeze-drying to obtain purified HOOC-PEOZ-OH powder;
step 2, dissolving the HOOC-PEOZ-OH powder obtained in the step 1 in dimethyl sulfoxide, adding acetic anhydride, magnetically stirring under the protection of nitrogen, after the reaction is finished, rotationally evaporating to remove the organic solvent in the reaction liquid, dropwise adding a small amount of dichloromethane for redissolution, precipitating with excessive cold ethyl ether, and performing suction filtration to obtain HOOC-PEOz-CHO solid powder;
step 3, dissolving chitosan in distilled water, and stirring until the chitosan is completely dissolved to prepare a solution I; dissolving the HOOC-PEOz-CHO solid powder prepared in the step 2 in absolute ethyl alcohol, dissolving NHS and EDC in absolute ethyl alcohol, pouring the absolute ethyl alcohol mixed solution containing HOOC-PEOz-CHO into the dissolved absolute ethyl alcohol mixed solution of NHS and EDC, stirring and dissolving at 65 ℃ to prepare a solution II; adding the solution II into the solution I under vigorous stirring, reacting at 65 ℃, putting the solution into a dialysis bag after reaction, dialyzing in distilled water, freeze-drying, washing the obtained product with absolute ethyl alcohol, and performing suction filtration to obtain CHO-PEOz-CS;
step 4, stirring and dissolving the CHO-PEOz-CS obtained in the step 3 and RGD in 95% ethanol with the pH of 4.6, and then dissolving NaBH3CN and CHO-PEOz-CS are dripped into the mixture according to a certain molar ratio, and the obtained product is dialyzed in distilled water and freeze-dried to obtain a product RGD-PEOz-CS;
and 5, precisely weighing RGD-PEOz-CS, dissolving the RGD-PEOz-CS in 1 acetate buffer solution, adding an equivalent volume of an antithrombotic drug solution, wherein the mass ratio of the antithrombotic drug to the RGD-PEOz-CS is 1: 2-10, adding a TPP solution with the concentration of 1-2 mg/mL into the solution under room-temperature magnetic stirring, and continuously stirring for 30-90 min after adding to obtain the drug-loaded RGD-PIC polyion micelle.
3. The preparation method of the thrombus-targeted long-circulating polycation micelle according to claim 2, wherein in the step 1, the mass ratio of the monomer EOz to the 3-bromo-ethyl propionate is 1-5: 1, the volume of acetonitrile is 80-250 mL, the molar ratio of the 3-bromo-ethyl propionate to the potassium iodide is 1: 0.5-2, the stirring speed is 300-800 rpm, the first heating reflux reaction time is 24-48 h, the temperature is 80-120 ℃, the molar ratio of the potassium hydroxide to the 3-bromo-ethyl propionate is 2-5: 1, the second heating reflux reaction time is 12-24 h, the temperature is 80-120 ℃, the rotary evaporation temperature is 40-80 ℃, the volume of dichloromethane is 10-20 mL, the volume of cold ether is 200-500 mL, the volume of water is 10-20 mL, the molecular weight cut-off of a dialysis bag is 8000-14000, and the dialysis time is 2-4 days, and (5) freeze-drying for 1-2 days.
4. The method for preparing a thrombus-targeted long-circulating polycation micelle according to claim 2, wherein in the step 2, HOOC-PEO is adoptedZThe mass of the-OH powder is 0.5-2 g, the volume of the dimethyl sulfoxide is 2-10 mL, the volume of the acetic anhydride is 0.1-0.5 mL, the stirring speed is 300-800 rpm, the stirring time is 8-24 h, the rotary evaporation temperature is 40-60 ℃, the volume of the dichloromethane is 10-20 mL, the volume of the cold ether is 200-500 mL, and the dialysis time is 2-4 days.
5. The method for preparing a thrombus-targeting long-circulating polycation micelle according to claim 2, wherein in the step 3, the mass of chitosan is 0.2-1 g, the volume of distilled water is 30-150 mL, the first stirring speed is 300-800 rpm, the mass of HOOC-PEOz-CHO is 0.1-0.5 g, the volume of absolute ethanol for dissolving HOOC-PEOz-CHO is 20-100 mL, the mass of NHS is 0.1-0.5 g, the mass of EDC is 0.1-0.5 g, the volume of absolute ethanol for dissolving NHS and EDC is 50-250 mL, the second stirring speed is 300-800 rpm, the reaction time is 3-6 h, the molecular weight of a dialysis bag is 8000-14000, the dialysis time is 2-4 days, and the freeze drying is 1-2 days.
6. The method for preparing a thrombus-targeting long-circulating polycation micelle according to claim 2, wherein the molar ratio of CHO-PEOz-CS to RGD in the step 4 is 1: 0.2-0.6, and the CHO-PEOz-CS and NaBH are used3CN is 1: 0.2-0.6, the cut-off molecular weight of the dialysis bag is 8000-14000, the dialysis time is 2-4 days, and the freeze drying is 1-2 days.
7. The method for preparing a thrombus-targeting long-circulating polycation micelle according to claim 2, wherein the antithrombotic agent in the step 5 is any one of heparin, natural hirudin, recombinant hirudin, streptokinase or urokinase.
8. The method for preparing a thrombus-targeting long-circulating polycation micelle according to claim 2, wherein the mass of RGD-PEOz-CS in the step 5 is 10-20 mg, the volume of acetate buffer solution is 10-20 mL, the stirring speed is 300-600 rpm, and the volume of TPP solution is 8-10 mL.
9. An application of a thrombus-targeted polyion micelle in the field of antithrombotic drug delivery.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101810559A (en) * | 2009-02-20 | 2010-08-25 | 北京大学 | Hirudin polyion micelle composition of targeted platelet |
| CN103319710A (en) * | 2013-06-06 | 2013-09-25 | 辽宁师范大学 | Poly(2-ethyl-2-oxazoline)-lipid derivatives and preparation method thereof |
| WO2014126537A1 (en) * | 2013-02-13 | 2014-08-21 | Agency For Science, Technology And Research | A Polymeric System for Release of an Active Agent |
| CN106983719A (en) * | 2017-03-08 | 2017-07-28 | 江苏富泽药业有限公司 | A kind of docetaxel polymer nano micelle injection, its preparation method and its application in tumor is prepared |
| CN110693851A (en) * | 2019-10-24 | 2020-01-17 | 西安医学院 | A kind of mesoporous silica drug-loaded nanoparticle and its preparation method and application |
-
2020
- 2020-11-11 CN CN202011254555.0A patent/CN112618487A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101810559A (en) * | 2009-02-20 | 2010-08-25 | 北京大学 | Hirudin polyion micelle composition of targeted platelet |
| WO2014126537A1 (en) * | 2013-02-13 | 2014-08-21 | Agency For Science, Technology And Research | A Polymeric System for Release of an Active Agent |
| CN103319710A (en) * | 2013-06-06 | 2013-09-25 | 辽宁师范大学 | Poly(2-ethyl-2-oxazoline)-lipid derivatives and preparation method thereof |
| CN106983719A (en) * | 2017-03-08 | 2017-07-28 | 江苏富泽药业有限公司 | A kind of docetaxel polymer nano micelle injection, its preparation method and its application in tumor is prepared |
| CN110693851A (en) * | 2019-10-24 | 2020-01-17 | 西安医学院 | A kind of mesoporous silica drug-loaded nanoparticle and its preparation method and application |
Non-Patent Citations (3)
| Title |
|---|
| LI ET AL.: "Poly(2-ethyl-2-oxazoline)-Doxorubicin Conjugate-Based Dual Endosomal pH-Sensitive Micelles with Enhanced Antitumor Efficacy", 《BIOCONJUGATE CHEMISTRY》 * |
| WANG ET AL.: "Nanoscaled Polyion Complex Micelles for Targeted Delivery of Recombinant Hirudin to Platelets Based on Cationic Copolymer", 《MOLECULAR PHARMACEUTICS》 * |
| 胡海洋 等: "pH敏感聚(2-乙基-2-噁唑啉)-壳聚糖-阿霉素共聚物胶束的制备及性质考察", 《沈阳药科大学学报》 * |
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