CN113616617A - Modified albumin nano particle capable of releasing NO gas by ultrasonic, preparation method and application thereof - Google Patents
Modified albumin nano particle capable of releasing NO gas by ultrasonic, preparation method and application thereof Download PDFInfo
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- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0002—Galenical forms characterised by the drug release technique; Application systems commanded by energy
- A61K9/0009—Galenical forms characterised by the drug release technique; Application systems commanded by energy involving or responsive to electricity, magnetism or acoustic waves; Galenical aspects of sonophoresis, iontophoresis, electroporation or electroosmosis
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- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0028—Disruption, e.g. by heat or ultrasounds, sonophysical or sonochemical activation, e.g. thermosensitive or heat-sensitive liposomes, disruption of calculi with a medicinal preparation and ultrasounds
- A61K41/0033—Sonodynamic cancer therapy with sonochemically active agents or sonosensitizers, having their cytotoxic effects enhanced through application of ultrasounds
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- 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
- A61K47/54—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 an organic compound
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
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- A61K9/5169—Proteins, e.g. albumin, gelatin
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
Abstract
The invention discloses a modified albumin nano particle capable of releasing NO gas by ultrasonic, a preparation method and application thereof. The modified albumin has SNO groups capable of effectively exciting and releasing NO by ultrasonic, NO reacts with superoxide anions to form peroxynitrite, the function of the albumin can be changed, DNA can be damaged, and the modified albumin is beneficial to killing tumor cells and intracellular pathogens by cooperating with chemotherapeutic drugs; the preparation method is characterized in that the desolvation method is utilized to avoid the toxicity of the drug to normal tissues, the nano-particle size distribution is uniform and controllable within the range of 20-500nm, the dispersibility is good, and the large-scale preparation is facilitated.
Description
Technical Field
The invention belongs to the field of new biomedical materials, and particularly relates to modified albumin nanoparticles capable of releasing NO signal molecules by ultrasound, a preparation method thereof, and application of the modified albumin nanoparticles in releasing medicaments by ultrasound.
Background
Albumin is a serum protein with the most abundant content in blood, and is often selected as a drug carrier due to its characteristics of high water solubility, good stability, high biocompatibility and the like, and a large number of amino acid residues exist in the primary structure of albumin, so that binding sites are provided for some small molecules (ligands), and the albumin can effectively load various different types of drugs. Human Serum Albumin (HSA) and Bovine Serum Albumin (BSA) have been the most studied carriers for drug delivery. HAS is a soluble single-chain globular protein with a molecular weight of 67kDa and consists of 585 amino acids, and HAS many drug binding sites, and can be bound with salicylic acid, warfarin, diazepam, penicillin and penicillin. BSA is a globular heart-shaped protein, has the molecular weight of 66kDa, and has the advantages of no toxicity, low antigenicity, biodegradability and the like. The BSA peptide chain consists of 583 amino acid residues, provides rich drug-loading sites, also contains 35 cysteine residues, one sulfydryl and the balance of disulfide bonds, and plays an important role in maintaining the spatial structure of albumin.
Chemotherapy is one of the indispensable important means for treating tumors, but the further clinical application of the chemotherapy is hindered by the problems of low target and large toxic and side effects in treatment. The nano-drug delivery system can effectively reduce the toxic and side effects of the drug, improve the solubility of the hydrophobic drug, improve the pharmacokinetics of the drug, enhance the passive targeting capability of the solid tumor through the EPR effect (high permeability and retention effect), and improve the enrichment of the drug at the tumor.
In a word, the prior art has the problem that the albumin nanoparticles can not controllably release the drugs under physiological conditions, and can only slowly release the drugs under the endogenous stimulation of acidity or reducibility and the like, and the defects of low drug release amount and the like caused by poor tissue penetrability of light-controlled release are urgently needed to be solved.
Although the chinese invention patent (application) CN202010186222.2 discloses an albumin nanoparticle capable of activating and releasing drugs by ultrasound, its preparation method and application, which uses azo cross-linking agent and encapsulates chemotherapeutic drugs for ultrasound release, the albumin used in this patent is unmodified albumin, and does not consider the modification of albumin, and does not involve the release of NO signal molecules.
Disclosure of Invention
The invention aims to provide modified albumin nano particles capable of releasing NO signal molecules through ultrasound, and a preparation method and application thereof. The nano particles have the characteristics of uniform and stable particle size distribution, and the modified albumin can release NO molecules with a tumor killing effect under the action of ultrasound, and the release amount of the NO molecules can be regulated and controlled according to the time of ultrasound.
In order to achieve the purpose, the invention obtains a group (NO donor structure) capable of releasing NO by ultrasonic through reducing part of disulfide bonds in an albumin structure and oxidizing, prepares modified albumin nano particles through a desolvation method, forms stable nano particles, can control the release amount of a medicament through the time and the intensity of ultrasonic, and achieves the purpose of quickly and controllably releasing NO molecules at a tumor position in vivo. Specifically, the following technical scheme is adopted:
The first aspect of the invention provides a preparation method of modified nanoparticles capable of releasing NO molecules by ultrasonic, which comprises the following steps:
(1) adding a disulfide bond reducing agent into the albumin aqueous solution, adjusting the pH value to 8-10, stirring and reacting for 12-48h at room temperature, adjusting the pH value to 3-5, and dialyzing for 2-48h with pure water to obtain a modified albumin solution with reduced disulfide bonds;
(2) adding a sulfhydryl oxidant into the modified albumin solution of the reduced disulfide bond obtained in the step (1), reacting for 2-24h at 4-50 ℃, and dialyzing for 2-48h with pure water to obtain a modified albumin solution containing-SNO groups capable of releasing NO gas;
(3) and (3) adjusting the pH value of the modified albumin solution containing-SNO groups capable of releasing NO gas obtained in the step (2) to 7-9, adding a certain proportion of organic solvent at a constant speed under stirring to perform a desolvation process to obtain a modified albumin nanoparticle suspension (modified albumin nanoparticle solution), and performing dehydration treatment to obtain modified albumin nanoparticles.
Preferably, in the step (1), the albumin molecule is one or a combination of two or more selected from human serum albumin, recombinant human serum albumin, bovine serum albumin, ovalbumin and animal serum albumin. Correspondingly, the obtained modified albumin nanoparticles are any one or a composition of more than two of modified human serum albumin nanoparticles, modified recombinant human serum albumin nanoparticles, modified bovine serum albumin nanoparticles, modified ovalbumin nanoparticles and modified animal serum albumin nanoparticles with an NO donor structure capable of releasing NO molecules by ultrasonic.
Preferably, in step (1), the concentration of albumin molecules in the aqueous solution is 1-200mg/mL, preferably 10-100 mg/mL; the disulfide bond reducing agent is 2-mercaptoethanol and dithiothreitol, and the concentration of the disulfide bond reducing agent is 10-100 mu L/mL; preferably, after adding the disulfide bond reducing agent, alkaline substances are used for adjusting the pH value to 8-10; the alkaline substance is one or a mixture of more than two of sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate and sodium bicarbonate, and the preferred concentration of the alkaline substance is 0.5-2 mol/L; preferably, after the stirring reaction, the pH value is adjusted to 3-5 by using an acidic substance; the acidic substance is one or more of hydrochloric acid, sulfuric acid and phosphoric acid, and the preferable concentration of the acidic substance is 0.5-2 mol/L. Preferably, the reaction time is 2-4h and the dialysis time is 12-24 h.
Preferably, in the step (2), the oxidizing agent is one or a mixture of two of tert-butyl nitrite and isopropyl nitrite, and the concentration of the sulfhydryl oxidizing agent is 3-10 μ L/mL; preferably, the reaction temperature is between room temperature and 37 ℃, and the reaction time is 10-16 h.
Preferably, in steps (1) and (2), the dialysis has a molecular weight cut-off of 500Da to 100 kDa; preferably 3000Da-12 kDa; the dialysis was performed at room temperature.
Preferably, in the step (3), the organic solvent is one or a mixture of two or more of methanol, ethanol, acetone, 1, 4-dioxane, dimethyl sulfoxide, n-propanol, isobutanol, polyethylene glycol and tetrahydrofuran; preferably, the rate of adding the organic solvent is 0.1-5mL/min, and more preferably the rate of adding the organic solvent is 1-4 mL/min; preferably, the volume ratio of the volume of the added organic solvent to the volume of the modified albumin aqueous solution containing the-SNO groups capable of releasing NO gas obtained in the step (2) is (0.1-10): 1; further preferred volume ratios are (0.5-3): 1; preferably, the pH value of the modified albumin solution containing the-SNO groups capable of releasing NO gas obtained in the step (2) is adjusted to 7-9 by using alkaline substances; the alkaline substance is one or more of sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate and sodium bicarbonate, and the preferred concentration of the alkaline substance is 0.1-2 mol/L.
Preferably, step (3) further comprises: and continuously reacting the obtained modified albumin nanoparticle suspension for 12-24h at 37 ℃ in a dark condition to obtain a stable modified albumin nanoparticle solution, and then dehydrating to obtain the stable albumin nanoparticles.
Preferably, in the step (3), the dehydration treatment is performed by high-speed centrifugation, freeze drying, spray drying, vacuum drying or distillation under reduced pressure.
The second aspect of the present invention provides modified albumin nanoparticles capable of ultrasonically releasing NO molecules, which are prepared by the method of the first aspect, and the modified albumin molecules capable of ultrasonically releasing NO molecules are used as a nanoparticle framework.
A third aspect of the invention provides the use of a nanoparticle according to the second aspect above for the release of NO molecules by ultrasound.
Preferably, the ultrasound used for the modified albumin nanoparticle drug that releases NO gas by ultrasound is within an ultrasonic power and ultrasonic frequency range acceptable for living organisms.
Preferably, the ultrasonic power is 0-3W/cm2More preferably 1.5 to 2.5W/cm2(ii) a Preferably, the ultrasonic frequency is 0.5 to 10MHz, more preferably 0.5 to 2 MHz.
NO is a unique diffusible messenger molecule that plays a central role in mammalian physiology. Its many effects include vascular smooth muscle relaxation, inhibition of platelet aggregation, effects on nerve signal transmission and modulation of immune function. In addition, NO reacts with superoxide anions (produced by activated macrophages and other cells) to form peroxynitrite. The by-product of NO is an effective chemical oxidant, altering albumin function and destroying DNA. These effects are part of the non-specific host defense, which helps to kill tumor cells and intracellular pathogens.
The invention has the beneficial effects that:
(1) the invention provides a modified albumin nanoparticle capable of releasing NO molecules by ultrasonic, wherein the modified albumin has SNO groups capable of effectively releasing NO by ultrasonic excitation, NO reacts with superoxide anions to form peroxynitrite, the function of albumin can be changed, DNA can be damaged, and the modified albumin nanoparticle is beneficial to killing tumor cells and intracellular pathogens by cooperating with chemotherapeutic drugs;
(2) the nano particles provided by the invention are prepared by a desolvation method, so that the toxicity of the medicine to normal tissues is avoided, the distribution of the nano particle size is uniform and controllable within the range of 20-500nm, the dispersibility is good, and the large-scale preparation is facilitated;
(3) the slow-release medicine ultrasound is used as an activation mode harmless to human bodies, compared with the traditional light-operated release mode, the slow-release medicine ultrasound has stronger tissue penetrability, can regulate and control the released NO gas through the ultrasound time, achieves the purpose of controllable release, and is beneficial to control.
Drawings
FIG. 1 is a UV-Vis spectrum of the modified albumin BSA-SH with reduced disulfide bonds prepared in example 1.
Fig. 2 is a transmission electron microscope image of modified albumin drug nanoparticles BSASNO capable of ultrasonically releasing NO gas prepared in example 3.
Fig. 3 shows the hydrated particle size of the modified albumin nanoparticle BSASNO nanoparticles (20ml ethanol) capable of releasing NO gas by ultrasound prepared in example 4.
Fig. 4 shows the hydrated particle size of the modified albumin nanoparticle BSASNO nanoparticles (15ml ethanol) capable of releasing NO gas by ultrasound prepared in example 5.
Fig. 5 is a graph of the change of NO gas released by BSASNO nanoparticles with ultrasound time.
Fig. 6 shows the hydrated particle size of the modified albumin BSASNO nanoparticles capable of releasing NO gas by ultrasound prepared in comparative example 1.
Detailed Description
The present invention will be described in detail below with reference to examples in order to make objects, technical solutions, and advantages of the present invention more apparent, but the present invention is not limited to the following examples.
Modified albumin nano particles capable of releasing NO gas by ultrasound are prepared by taking modified albumin molecules containing groups capable of releasing NO gas by ultrasound as a nano particle framework through a desolvation method, and are activated to release NO gas controllably under the action of ultrasound, and the release amount of NO is related to the time of ultrasound.
Example 1
Preparation of modified albumin with reduced disulfide bonds
5g of bovine serum albumin powder was dissolved in 100g of ultrapure water, and the mixture was stirred by a magnetic stirrer for 1 hour at 200 rpm. 6.975g of albumin aqueous solution was taken, 284. mu.l of 2-mercaptoethanol was added, the pH of the solution was adjusted to 8 with 1M NaOH, and the reaction was stirred at room temperature overnight. Adjusting the pH value of the solution to 3 by using 1M HCl, dialyzing for 24h by using a dialysis bag with the molecular weight cutoff of 3500Da, and adjusting the pH value to 5 in the dialysis process to obtain a modified albumin BSA-SH solution with reduced disulfide bonds.
The obtained modified bovine serum albumin is characterized in that as shown in figure 1, free sulfydryl obtained by reducing disulfide bonds in the modified bovine serum albumin reacts with a total sulfydryl reagent, and an absorption peak exists at the wavelength of 420nm in the ultraviolet visible spectrum.
Example 2
Preparation of modified albumin containing-SNO group capable of releasing NO gas
5mL of the BSA-SH solution prepared in example 1 was taken, 37. mu.L of tert-butyl nitrite was added thereto, the mixture was stirred at room temperature for 12 hours, and dialyzed with a dialysis bag having a cut-off molecular weight of 3500Da for 24 hours to obtain a BSA-SNO solution of modified albumin containing-SNO groups capable of releasing NO gas.
Example 3
Modified bovine serum albumin based nanoparticles (BSASNO) were prepared by the desolvation method. In principle, 5mL of the BSA-SNO solution prepared in example 2 was measured, the pH was adjusted to 7 with 0.1mol/L NaOH, 2.5mL of absolute ethanol was added dropwise at a constant rate of 2.5mL/min, and after the desolvation process was completed, the reaction was continued overnight at 37 ℃ in the dark to obtain a stable modified albumin nanoparticle solution. And centrifugally separating the obtained stable modified albumin nano solution at 12000rpm, washing the precipitate with deionized water, repeating the steps for three times, dispersing with a certain amount of deionized water, and freeze-drying to obtain the modified bovine serum albumin BSASNO nano particles.
The transmission electron microscope image of the obtained modified albumin BSASNO nano particles capable of releasing NO gas by ultrasonic is shown in figure 2, and the BSASNO nano particles have the radius of about 30nm and are uniformly distributed.
Example 4
Modified bovine serum albumin based nanoparticles (BSASNO) were prepared by the desolvation method. In principle, 5mL of the BSA-SNO solution prepared in example 2 was measured, the pH was adjusted to 9 with 0.1mol/L NaOH, 8mL of absolute ethanol was added dropwise at a constant rate of 4mL/min, and after the desolvation process was completed, the reaction was continued overnight at 37 ℃ in the dark to obtain a stable modified albumin nanoparticle solution. And centrifugally separating the obtained stable modified albumin nano solution at 12000rpm, washing the precipitate with deionized water, repeating the steps for three times, dispersing with a certain amount of deionized water, and freeze-drying to obtain the modified bovine serum albumin BSASNO nano particles.
The hydration particle size of the obtained modified albumin BSASNO nano particle capable of releasing NO gas by ultrasonic is shown in figure 3, and the hydration particle size of the BSASNO nano particle is 300 +/-20 nm.
Example 5
Modified bovine serum albumin based nanoparticles (BSASNO) were prepared by the desolvation method. In principle, 5mL of the BSA-SNO solution prepared in example 2 was measured, the pH was adjusted to 9, 6mL of absolute ethanol was added dropwise at a constant rate of 3mL/min, and after the desolvation process was completed, the reaction was continued overnight at 37 ℃ in the dark to obtain a stable modified albumin nanoparticle solution. And centrifugally separating the obtained stable modified albumin nano solution at 12000rpm, washing the precipitate with deionized water, repeating the steps for three times, dispersing with a certain amount of deionized water, and freeze-drying to obtain the modified bovine serum albumin BSASNO nano particles.
The hydration particle size of the obtained modified albumin BSASNO nano particle capable of releasing NO gas by ultrasonic is shown in figure 4, and the hydration particle size of the BSASNO nano particle is 500 +/-50 nm.
Example 6
Release of NO gas by modified albumin nanoparticles (BSA-SNO) at different ultrasound times
50ul of the BSASNO nanoparticle solution prepared in example 2 was dispersed in 3ml of ultrapure water, placed in a six-well plate, and the space between the six-well plate and an ultrasonic machine was filled with ultrapure water at 2MHz and 2.0W/cm2Selecting different times (0,1.0,2.0,3.0,4.0 and 5.0min) for ultrasonic treatment under the ultrasonic condition, transferring the nanoparticle solution in the six-hole plate to a four-side dish after the ultrasonic treatment is finished, and measuring the absorbance at 540 nm.
The curve of the change of the NO gas released by the BSASNO nanoparticles with the ultrasonic time as shown in fig. 5 shows that the NO gas released by the BSASNO nanoparticles with the ultrasonic time increases, and the speed does not show a significant decrease trend with the increase of the time.
Comparative example 1
Modified bovine serum albumin based nanoparticles (BSASNO) were prepared by the desolvation method. In principle, 5mL of BSA-SNO solution prepared in example 2 was measured, 2mL of absolute ethanol was added dropwise at a constant rate of 2mL/min, and after the desolvation process was completed, the reaction was continued overnight at 37 ℃ in the dark to obtain a stable modified albumin nanoparticle solution. And centrifugally separating the obtained modified albumin nano suspension at 12000rpm, washing the precipitate with deionized water, repeating the steps for three times, dispersing with a certain amount of deionized water, and freeze-drying to obtain the modified bovine serum albumin nano particles.
The hydration particle size of the obtained modified albumin BSASNO nano particle capable of releasing NO gas by ultrasonic is shown in figure 6, and it can be seen that the BSASNO nano particle has serious coagulation phenomenon and is not uniform.
The applicant states that the product and the detailed preparation method of the present invention are illustrated by the above examples, but the present invention is not limited to the above product and the detailed preparation method, i.e. the present invention is not meant to be implemented by relying on the above product and the detailed preparation method. Persons skilled in the art should understand that the persons and improvements of the present invention, the equivalent substitution of each raw material and the addition of auxiliary components, the selection of specific modes, etc. of the product of the present invention all fall within the protection scope and the disclosure scope of the present invention.
Claims (10)
1. A preparation method of modified nanoparticles capable of releasing NO molecules by ultrasonic is characterized by comprising the following steps:
(1) adding a disulfide bond reducing agent into the albumin aqueous solution, adjusting the pH value to 8-10, stirring and reacting for 12-48h at room temperature, adjusting the pH value to 3-5, and dialyzing for 2-48h with pure water to obtain a modified albumin solution with reduced disulfide bonds;
(2) adding a sulfhydryl oxidant into the modified albumin solution of the reduced disulfide bond obtained in the step (1), reacting for 2-24h at 4-50 ℃, and dialyzing for 2-48h with pure water to obtain a modified albumin solution containing-SNO groups capable of releasing NO gas;
(3) And (3) adjusting the pH value of the modified albumin solution containing-SNO groups capable of releasing NO gas obtained in the step (2) to 7-9, adding an organic solvent under stirring to perform a desolvation process to obtain a modified albumin nanoparticle suspension, and performing dehydration treatment to obtain modified albumin nanoparticles.
2. The method for preparing the modified nanoparticle capable of ultrasonically releasing NO molecules according to claim 1, wherein in the step (1), the albumin molecules are any one or a combination of more than two of human serum albumin, recombinant human serum albumin, bovine serum albumin, ovalbumin and animal serum albumin; the concentration of albumin in the water solution is 1-200mg/mL, preferably 10-100 mg/mL; the disulfide bond reducing agent is 2-mercaptoethanol and dithiothreitol, and the concentration of the disulfide bond reducing agent is 10-100 mu L/mL; the reaction time is 2-4h, and the dialysis time is 12-24 h.
3. The method for preparing modified nanoparticles capable of ultrasonically releasing NO molecules according to claim 1, wherein in the step (2), the oxidizing agent is one or a mixture of two of tert-butyl nitrite and isopropyl nitrite, and the concentration of the sulfhydryl oxidizing agent is 3-10 μ L/mL; the reaction temperature is between room temperature and 37 ℃, and the reaction time is 10-16 h.
4. The method for preparing modified nanoparticles capable of releasing NO molecules ultrasonically according to claim 1, wherein in the step (3), the organic solvent is one or a mixture of two or more of methanol, ethanol, acetone, 1, 4-dioxane, dimethyl sulfoxide, n-propanol, isobutanol, polyethylene glycol and tetrahydrofuran; the rate of adding the organic solvent is 0.1-5mL/min, and the rate of adding the organic solvent is 1-4 mL/min; the volume ratio of the added organic solvent to the aqueous solution of modified albumin containing-SNO groups which can release NO gas obtained in step (2) is (0.1-10):1, and more preferably (0.5-3): 1.
5. The method for preparing modified nanoparticles capable of ultrasonically releasing NO molecules according to claim 1, wherein in the steps (1) and (2), the dialysis has a molecular weight cut-off of 500Da to 100kDa, preferably 3000Da to 12 kDa;
in the step (3), the dehydration treatment method is high-speed centrifugation, freeze drying, spray drying, vacuum drying or reduced pressure distillation.
6. The method for preparing modified nanoparticles capable of ultrasonically releasing NO molecules according to claim 1, wherein in the step (1), after the disulfide bond reducing agent is added, the pH value is adjusted to 8-10 by using an alkaline substance; after stirring and reacting, adjusting the pH value to 3-5 by using an acidic substance; in the step (3), the pH value of the modified albumin solution containing the-SNO group capable of releasing NO gas obtained in the step (2) is adjusted to 7-9 by using alkaline substances;
Preferably, the alkaline substance used is one or a mixture of more than two of sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate and sodium bicarbonate; the acidic substance is one or a mixture of more than two of hydrochloric acid, sulfuric acid and phosphoric acid;
preferably, step (3) further comprises: and continuously reacting the obtained modified albumin nanoparticle suspension for 12-24h at 37 ℃ in a dark condition to obtain a stable modified albumin nanoparticle solution, and then dehydrating to obtain the stable albumin nanoparticles.
7. Modified albumin nanoparticles ultrasonically releasable of NO molecules prepared by the method of any one of claims 1-6.
8. Use of modified albumin nanoparticles according to claim 7 for the release of NO molecules by ultrasound.
9. Use according to claim 8, wherein the ultrasound means is in a range of acceptable ultrasound power and ultrasound frequency for the living organism.
10. Use according to claim 8 or 9, wherein the ultrasonic power is 0-3W/cm2More preferably 1.5 to 2.5W/cm2(ii) a The ultrasonic frequency is 0.5-10MHz, and more preferably 0.5-2 MHz.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114376067A (en) * | 2022-01-18 | 2022-04-22 | 大连工业大学 | Modified protein nano-particle, preparation method and application thereof |
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| US20130184237A1 (en) * | 2010-07-09 | 2013-07-18 | Fresenius Kabi Deutschland Gmbh | Nitric oxide delivering hydroxyalkyl starch derivatives |
| CN111407740A (en) * | 2020-03-17 | 2020-07-14 | 大连理工大学 | Albumin nanoparticles capable of activating and releasing drugs by ultrasound, and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114376067A (en) * | 2022-01-18 | 2022-04-22 | 大连工业大学 | Modified protein nano-particle, preparation method and application thereof |
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Application publication date: 20211109 |