CN112704741A - Preparation and application of serum albumin-non-steroidal anti-inflammatory drug nano preparation - Google Patents
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Abstract
The invention name is as follows: the preparation and application technical field of a serum albumin-non-steroidal anti-inflammatory drug nano preparation: summary content of advanced nano medicine and technical field: the invention introduces a preparation and application of a serum albumin-non-steroidal anti-inflammatory drug nano preparation formed based on a self-assembly technology, which comprises the following specific steps: the serum albumin and the non-steroidal anti-inflammatory drug or the mixed solution of the serum albumin, the trivalent cerium salt and the non-steroidal anti-inflammatory drug are stirred and then purified to obtain the serum albumin-non-steroidal anti-inflammatory drug. The preparation method is simple and rapid, green and environment-friendly, the synthetic raw materials are cheap and easy to obtain, and mass production can be realized. The bovine or human serum albumin can be combined with any one of the non-steroidal anti-inflammatory drugs to obtain a nano preparation. The nano preparation can improve the water solubility, the drug effect and the side effect of the non-steroidal anti-inflammatory drug, and can realize the secondary loading and the visual tracking of the therapeutic drug; secondly, the compound can be combined with any one or more excipients or auxiliary materials to obtain a corresponding pharmaceutical dosage form or a new medicine, so as to have the function of efficiently and synergistically treating diseases, in particular various acute and chronic inflammatory diseases, by using the non-steroidal anti-inflammatory drug or/and the cerium nano particles.
Description
Technical Field
The invention belongs to the field of advanced nano medicine and technology, relates to a preparation method of a nano material for treating inflammatory diseases, and also relates to a product prepared by the method and application, in particular to a serum albumin-non-steroidal anti-inflammatory drug nano preparation formed based on a self-assembly technology.
Background
Inflammation is a defensive response of living tissue with a vascular system to an injury factor. Inflammation is a double sword, and in general, inflammation is beneficial to the body, but excessive inflammatory response is potentially harmful. Inflammatory reactions are the basis of the onset of a large number of diseases, and when inflammation is too severe, it can threaten human life, such as severe hypersensitivity. In addition, inflammation occurring at specific sites or organs can have serious consequences, such as systemic degenerative osteoarthritis can lead to disability; inflammation of the brain may stress the nerve center; severe myocarditis can affect cardiac function. In the inflammation process, there are both injuries and anti-injuries, and the extent of the reactions between them determines the occurrence, development and outcome of inflammation. If the injury process is dominant, the inflammation is aggravated and spreads to the whole body; on the contrary, if the anti-injury reaction is dominant, the inflammation tends to be healed gradually; if the injury factor persists, or the body is less resistant, the inflammation becomes chronic. Thus, inflammation is one of the medical problems of the world that is not negligible, as little as cold fever, as much as tumor cancer, all with an inflammatory response.
The most commonly used drugs in inflammatory diseases are non-steroidal anti-inflammatory drugs, which are a class of anti-inflammatory drugs that do not contain a steroidal structure. First-line non-steroidal anti-inflammatory drugs include aspirin, indomethacin, sulindac, and the like, wherein indomethacin drug substance is soluble in acetone, slightly soluble in methanol, ethanol, chloroform or diethyl ether, very slightly soluble in toluene, and hardly soluble in water. The medicine is clinically orally taken and used for symptoms such as fever, pain and the like, but the medicine has more side effects such as gastrointestinal tract reaction, central nervous system reaction, hematopoietic system reaction, skin and anaphylactic reaction, irritation caused by local eye drop, burning sensation, liver toxicity, renal toxicity and the like. The sulindac belongs to a prodrug without pharmacological activity, and has lasting anti-inflammatory, analgesic and antipyretic effects after being reduced into sulfide by liver metabolism or intestinal flora. Sulindac can be metabolized into active sulfide and inactive sulfone, and sulfide and proto-drug can be mutually converted, wherein the half-life period of the proto-drug is 7.8h, and the half-life period of the active metabolite is 14 h. The drug is eventually excreted through the feces and urine as a crude drug or as an inactive metabolite or glucuronic acid conjugate. Its clinical indications include proliferative osteoarthropathy, rheumatoid arthritis; pain from various causes; light and moderate cancer pain, etc., it is required to be taken orally. Side effects are minor, but gastrointestinal reactions are the most common adverse reactions; it also has less damage to the central nervous system and kidney than other nsaids, but it also causes liver damage. It is very slightly soluble in water and is a hydrophobic drug.
Therefore, how to effectively improve the water solubility and the side effect of the non-steroidal anti-inflammatory drug on the premise of ensuring the drug effect of the non-steroidal anti-inflammatory drug is a problem to be solved at present. There are studies reporting that there is interaction between nsaids and serum albumin, mainly in the sub-region of the 1-site of serum albumin. Bovine serum albumin, a globulin protein in bovine serum, contains 607 amino acid residues, has a molecular weight of 66.446KD, and has a thiol group in its structure, thereby having reducibility. In addition, human serum albumin is an approved drug by the FDA, is a protein in human plasma, has a molecular weight of 66KD, and also has reducibility. Human serum albumin can transport fatty acids, bile pigments, amino acids and many therapeutic molecules etc. in body fluids while maintaining normal osmotic pressure of blood; in clinic, human serum albumin can be used for treating shock and burn, supplementing blood loss caused by operation, accident or hemorrhage, and can also be used as blood plasma bulking agent. In addition, bovine/human serum albumin can bind to various cations, anions and other small molecular substances, and mainly plays roles in maintaining osmotic pressure, pH buffering, carrier and nutrition. Notably, sulindac in nsaids can be reduced to active ingredients by reducing substances to exert direct pharmacological effects. The concentration of the bovine/human serum albumin under physiological conditions is 40mg/mL, so that the bovine/human serum albumin and the non-steroidal anti-inflammatory drug under physiological conditions are combined and applied through non-chemical reaction to prepare the bovine/human serum albumin-non-steroidal anti-inflammatory drug nano preparation with special structure and effect, thereby removing the toxic cosolvent of the traditional non-steroidal anti-inflammatory drug, improving the safe dose of the drug, and being expected to produce better pharmacological effect and substantial clinical improvement.
In recent years, cerium nanoparticles are an inorganic nanomaterial with unique properties, and the trivalent state and the tetravalent state of cerium coexist on the surface of the inorganic nanomaterial and are mutually converted. Based on different valence state ratios, the compounds have strong mimic enzyme activities, such as superoxide enzyme, catalase, oxidative phosphatase and the like, so that various free radicals can be efficiently and permanently eliminated. Meanwhile, the oxidation resistance of the ultra-small cerium oxide nano particles can remove active oxygen at inflammatory parts, so that the ultra-small cerium oxide nano particles have very important and wide application prospects in the directions of chemical catalytic reaction, radiation protection, tumor radiotherapy and chemotherapy, treatment of oxidation-resistant diseases, regenerative medicine, inflammatory diseases and the like. However, the cerium nanoparticles have poor stability, and the problems of different degrees of biological activity and biocompatibility exist, so that the research and analysis results of the cerium nanoparticles can be influenced to a certain extent, so that the development of more active cerium nano materials is a relatively advanced research at present, and compared with the pure cerium nanoparticles, the cerium-mediated serum albumin-nonsteroidal anti-inflammatory drug nano preparation has more excellent activity and more biological application prospect.
Serum albumin is taken as a carrier, the serum albumin, a conventional non-steroidal anti-inflammatory drug and a cerium material are prepared into a nano preparation through a self-assembly technology, the water solubility and the oral absorption of the non-steroidal anti-inflammatory drug are improved, the internal circulation time is increased, the bioavailability is improved, the targeting effect is achieved on an action part, the drug dosage is reduced, and the side effect is reduced; meanwhile, serum albumin is discharged through biological metabolism, and has no toxic or side effect. The bovine or human serum albumin and the non-steroidal anti-inflammatory drug are prepared into the nano preparation for the first time and are applied, and the developed nano preparation is proved to have good application prospect in clinical research.
Disclosure of Invention
In view of the above, aiming at the key problem that chemical stability, biological activity and biocompatibility are difficult to be effectively ensured in the synthesis of the existing nano material, a preparation method of a serum albumin-non-steroidal anti-inflammatory drug nano preparation is established, the synthesis of the nano preparation is realized through one-step self-assembly, a simple, green, economical and practical synthesis method is provided, and a technical guarantee is provided for further application of a protein nano material in the field of biomedicine; the second purpose of the invention is to provide a serum albumin-non-steroidal anti-inflammatory drug nano preparation prepared by the method; the invention also aims to provide the application of the obtained serum albumin-non-steroidal anti-inflammatory drug nano preparation, provide a new idea for developing a novel nano drug delivery platform and provide a new scheme for treating patients with clinical inflammatory diseases.
In order to achieve the purpose, the invention provides the following technical scheme:
1. a method for preparing serum albumin-NSAID nanometer preparation, the process and the schematic diagram are shown in figure 1, comprising the following steps:
combining serum albumin with a nonsteroidal anti-inflammatory drug: dissolving serum albumin in water, adding the non-steroidal anti-inflammatory drug, carrying out ice-bath reaction for 24h under magnetic stirring, and purifying to obtain the serum albumin-non-steroidal anti-inflammatory drug nano preparation. (iii) cerium-mediated serum albumin in combination with a non-steroidal anti-inflammatory drug: dissolving serum albumin in water, adding trivalent cerium salt, reacting for 15min under magnetic stirring to synthesize a precursor, adding an alkaline solution into a reaction system, carrying out nucleation and curing reaction, adding a non-steroidal anti-inflammatory drug, and continuing to react for 15min, wherein the reaction system is always carried out in a nitrogen atmosphere. Purifying to remove unreacted inorganic ions and unreacted protein to obtain the cerium-mediated serum albumin-non-steroidal anti-inflammatory drug nano preparation.
Preferably, the serum albumin is bovine serum albumin, human serum albumin.
Preferably, the non-steroidal anti-inflammatory drug is an optional non-steroidal anti-inflammatory drug, such as sulindac, indomethacin, ibuprofen, or aspirin.
Preferably, the trivalent cerium salt is cerium nitrate, cerium trichloride or cerium sulfate; the alkaline solution is sodium hydroxide, potassium hydroxide or ammonia water.
Preferably, there is a defined ratio in composition between the serum albumin-nsaid nanoformulation and the cerium mediated serum albumin-nsaid nanoformulation. The molar ratio of the serum albumin to the non-steroidal anti-inflammatory drug is 0.1-1.5: 1-10, wherein the molar ratio of serum albumin, trivalent cerium salt, alkaline solution and non-steroidal anti-inflammatory drug is 0.1-1.5: 1-10: 10-100: 0.1 to 1.5.
More preferably, the nucleation curing reaction condition is that the reaction is carried out for 10min to 72h at the temperature of 4 to 80 ℃; the magnetic stirring is carried out at 500-1400 rpm/min.
2. The serum albumin and the nonsteroidal anti-inflammatory drug and the cerium-mediated serum albumin and the nonsteroidal anti-inflammatory drug are combined by a special chemical bond, and the preparation is a novel nano preparation.
3. The serum albumin-nonsteroidal anti-inflammatory drug nano preparation and the cerium-mediated serum albumin-nonsteroidal anti-inflammatory drug nano preparation described by the invention can be combined with one or more optional excipients or auxiliary materials to obtain corresponding drug formulations or new drugs. The pharmaceutical dosage form or new drug can be prepared into various other existing dosage forms according to the purpose, direction and the like required to be achieved, and can be prepared by methods known in the art.
4. The infrared spectrum of the serum albumin-nonsteroidal anti-inflammatory drug nano preparation and the cerium-mediated serum albumin-nonsteroidal anti-inflammatory drug nano preparation is different from that of a physical mixture of the serum albumin, the nonsteroidal anti-inflammatory drug, the serum albumin and the nonsteroidal anti-inflammatory drug.
5. The serum albumin-non-steroidal anti-inflammatory drug nano preparation and the cerium-mediated serum albumin-non-steroidal anti-inflammatory drug nano preparation have the function of treating diseases by using non-steroidal anti-inflammatory drugs or/and cerium nano particles, and the obtained serum albumin-non-steroidal anti-inflammatory drug nano preparation has a great promoting effect and an application prospect in the fields of drug targeted delivery, treatment of acute and chronic inflammatory diseases and regenerative medicine.
The invention has the beneficial effects that: the invention discloses a method based on a serum albumin-non-steroidal anti-inflammatory drug nano preparation, on one hand, the synthesis of the serum albumin-non-steroidal anti-inflammatory drug nano preparation is successfully realized through simple regulation and control of a system, the nano preparation does not need further modification, and the stability and the biocompatibility are correspondingly guaranteed; on the other hand, the cerium mediated serum albumin-non-steroidal anti-inflammatory drug nano preparation has excellent biological compatibility and enzymatic activity, improves the focus environment, and lays a foundation for further application in organisms. Finally, the method is environment-friendly and economical in reaction, flexible and easy to control in regulation and control mode and strong in repeatability.
Drawings
In order to make the object, technical scheme and beneficial effect of the invention more clear, the invention provides the following drawings for explanation:
FIG. 1 is a schematic diagram of the construction of the bovine/human serum albumin and non-steroidal anti-inflammatory drug sulindac and cerium-mediated bovine/human serum albumin and non-steroidal anti-inflammatory drug sulindac nano-preparation of the present invention.
FIG. 2 is a picture, ultraviolet spectrum and infrared spectrum of a product of a nano preparation formed by bovine serum albumin and a non-steroidal anti-inflammatory drug sulindac, cerium-mediated bovine serum albumin and a non-steroidal anti-inflammatory drug sulindac.
FIG. 3 is a diagram of the hydrated particle size and transmission electron microscope of the nanometer preparation of the bovine serum albumin and the non-steroidal anti-inflammatory drug sulindac.
Figure 4 is an atomic force microscope image of a cerium-mediated bovine/human serum albumin and a non-steroidal anti-inflammatory drug sulindac formed into a nano-formulation according to the present invention.
FIG. 5 is a transmission electron microscope image of the cerium mediated bovine/human serum albumin and the non-steroidal anti-inflammatory drug sulindac formed into the nano-preparation of the present invention.
FIG. 6 is the potential diagram of the nano preparation formed by bovine serum albumin and non-steroidal anti-inflammatory drug sulindac, cerium mediated bovine/human serum albumin and non-steroidal anti-inflammatory drug sulindac.
FIG. 7 is a molecular simulation diagram of bovine serum albumin and the NSAID sulindac of the present invention.
FIG. 8 is a molecular mimetic diagram of the cerium-mediated bovine/human serum albumin and the non-steroidal anti-inflammatory drug sulindac of the present invention.
Detailed Description
The above-described aspects of the present invention are explained in further detail by the following embodiments in specific forms, but they are not to be construed as limiting the scope of the present invention. Given the above, it is within the scope of the invention to extend the scope of the invention as claimed to the extent available to those skilled in the art to which the invention pertains.
Example 1 preparation of bovine serum Albumin-Indometacin Nanodimulants
Taking 20mL of 40mg/mL bovine serum albumin aqueous solution, slowly dropwise adding 50 mu L of 200mg/mL indomethacin dimethyl sulfoxide solution under magnetic stirring at 600rpm/min, and stirring for 24h at 4-10 ℃. At this time, the amount of dimethyl sulfoxide in the reaction system is two thousandth, and the dimethyl sulfoxide is nontoxic to cell animals. Filtering the reaction solution with a water-soluble microporous filter membrane of 0.22 mu m after the reaction is finished, then centrifugally washing the reaction solution for 5min by a 100KD jacketed centrifuge tube at 5000rpm to remove redundant unreacted bovine serum albumin and indometacin, washing the reaction solution for 2 times, and then fixing the volume to 10mL by using ultrapure water to obtain the bovine serum albumin-indometacin nano preparation which is marked and stored at 4 ℃ for later use.
Example 2 preparation of human serum Albumin-Indometacin Nanodimulations
20mL of 40mg/mL human serum albumin aqueous solution is taken, 50 mu L of 200mg/mL indomethacin dimethyl sulfoxide solution is slowly dripped under magnetic stirring at 600rpm/min, and the mixture is stirred for 24 hours at the temperature of 4-10 ℃. At this time, the amount of dimethyl sulfoxide in the reaction system is two thousandth, and the dimethyl sulfoxide is nontoxic to cell animals. Filtering the reaction solution with a water-soluble microporous filter membrane of 0.22 μm after the reaction is finished, then centrifugally washing the reaction solution with a 100KD jacketed centrifuge tube at 5000rpm for 5min to remove redundant unreacted human serum albumin and indometacin, washing the reaction solution for 2 times, and then fixing the volume to 10mL with ultrapure water to obtain a human serum albumin-indometacin nano preparation which is marked and stored at 4 ℃ for later use.
Example 3 preparation of bovine serum Albumin-sulindac Nanodiulation
Taking 20mL of 40mg/mL bovine serum albumin aqueous solution, slowly dropwise adding 50 mu L of 200mg/mL sulindac dimethyl sulfoxide solution under magnetic stirring at 600rpm/min, and stirring for 24h at 4-10 ℃. At this time, the amount of dimethyl sulfoxide in the reaction system is two thousandth, and the dimethyl sulfoxide is nontoxic to cell animals. Filtering the reaction solution with a water-soluble microporous filter membrane of 0.22 mu m after the reaction is finished, then centrifugally washing the reaction solution for 5min by a 100KD jacketed centrifuge tube at 5000rpm to remove redundant unreacted bovine serum albumin and sulindac, washing the reaction solution for 2 times, and then fixing the volume to 10mL by using ultrapure water to obtain a bovine serum albumin-sulindac nano preparation which is marked and stored at 4 ℃ for later use. The product state is shown in figure 2a, which obviously improves the water solubility of sulindac bulk drug; further using an ultraviolet-visible spectrophotometer to verify the success of the material synthesis (fig. 2 b); the Fourier infrared spectrum proves that the material has the characteristic peaks of pure bovine serum albumin and sulindac, but the characteristic peaks are different from the characteristic peaks of a mixture of the bovine serum albumin and the sulindac (figure 2 c); the morphology of the compound is characterized by a hydrated particle size diagram and a transmission electron microscope diagram, as shown in figure 3, the size of a nano preparation formed by bovine serum albumin and a non-steroidal anti-inflammatory drug sulindac is about 10nm, and the nano preparation is in a uniform spherical shape; the zeta potential is shown in FIG. 6 and is about-27 mV, indicating good stability. In addition, the successful docking of bovine serum albumin and sulindac was simulated by Drug Studio software (fig. 7). Therefore, the bovine serum albumin-sulindac nano preparation is successfully prepared.
Example 4 preparation of human serum Albumin-sulindac Nanodiulation
Taking 20mL of 40mg/mL human serum albumin aqueous solution, slowly dropwise adding 50 mu L of 200mg/mL sulindac dimethyl sulfoxide solution under magnetic stirring at 600rpm/min, and stirring for 24h at 4-10 ℃. At this time, the amount of dimethyl sulfoxide in the reaction system is two thousandth, and the dimethyl sulfoxide is nontoxic to cell animals. Filtering the reaction solution with a water-soluble microporous filter membrane of 0.22 μm after the reaction is finished, then centrifugally washing the reaction solution for 5min by a 100KD jacketed centrifuge tube at 5000rpm to remove redundant unreacted human serum albumin and sulindac, washing for 2 times, and then fixing the volume to 10mL by ultrapure water to obtain a human serum albumin-sulindac nano preparation which is marked and stored at 4 ℃ for later use.
Example 5 preparation of cerium-mediated bovine serum Albumin-sulindac Nanodiulation
9mL of 25mg/mL bovine serum albumin aqueous solution is taken, one end of a three-neck flask is connected with a balloon filled with nitrogen by a three-port adapter, air in the three-neck flask is pumped by a vacuum pump and filled with nitrogen, 0.5mL of 0.1mol/L cerium nitrate hexahydrate aqueous solution is added by an injector, the mixture is stirred for 15min at room temperature, 0.6mL of 1mol/L potassium hydroxide aqueous solution is quickly dripped by the injector, 100 mu L of 200mg/mL sulindac is dripped by the injector continuously, and the stirring reaction is continued for 15 min. And (3) after the reaction is finished, carrying out ultrapure water dialysis and purification for 2h, replacing ultrapure water once per hour by using a snakeskin dialysis bag with the molecular weight cutoff of 10KD during dialysis, and finally collecting liquid in the dialysis bag to obtain a cerium-mediated bovine serum albumin-nonsteroidal anti-inflammatory drug nano preparation which is marked and stored at 4 ℃ for later use. The product state is shown in figure 2a, which obviously improves the water solubility of sulindac bulk drug; further preliminarily verifying the success of drug loading by using an ultraviolet-visible spectrophotometer (fig. 2 b); fourier infrared spectrum proves that the material has the characteristic peaks of pure bovine serum albumin and sulindac, but the characteristic peaks are different from the characteristic peaks of a mixture of the bovine serum albumin, the sulindac and cerium (figure 2 c); the morphology of the cerium-mediated bovine serum albumin-sulindac nano preparation is represented by an atomic force microscope and a transmission electron microscope image, and as shown in fig. 4 and 5, the size of the cerium-mediated bovine serum albumin-sulindac nano preparation is about 8nm and is in a uniform spherical shape. The zeta potential is shown in FIG. 6 and is about-43.67 mV, indicating good stability. In addition, the successful docking of bovine serum albumin and sulindac was simulated by Drug Studio software (fig. 8). Therefore, the invention successfully prepares the cerium-mediated bovine serum albumin-sulindac nano preparation.
Example 6 preparation of cerium-mediated human serum Albumin-sulindac Nanodiulation
9mL of 25mg/mL human serum albumin aqueous solution is taken, one end of a three-neck flask is connected with a balloon filled with nitrogen by a three-port adapter, air in the three-neck flask is pumped by a vacuum pump and filled with nitrogen, 0.5mL of 0.1mol/L cerium nitrate hexahydrate aqueous solution is added by an injector, the mixture is stirred for 15min at room temperature, 0.6mL of 1mol/L potassium hydroxide aqueous solution is quickly dripped by the injector, 100 mu L of 200mg/mL sulindac is dripped by the injector continuously, and the stirring reaction is continued for 15 min. And (3) after the reaction is finished, carrying out ultrapure water dialysis and purification for 2h, replacing ultrapure water once per hour by using a snakeskin dialysis bag with the molecular weight cutoff of 10kD during dialysis, and finally collecting liquid in the dialysis bag to obtain a cerium-mediated human serum albumin-nonsteroidal anti-inflammatory drug nano preparation, wherein the nano preparation is marked and stored at 4 ℃ for later use.
Claims (10)
1. A method for preparing a serum albumin-non-steroidal anti-inflammatory drug nano preparation is characterized by comprising the following steps: serum albumin in combination with non-steroidal anti-inflammatory drugs: dissolving serum albumin in water, adding the non-steroidal anti-inflammatory drug, carrying out ice-bath reaction for 24h under magnetic stirring, and purifying to obtain the serum albumin-non-steroidal anti-inflammatory drug nano preparation.
2. A method for preparing a serum albumin-non-steroidal anti-inflammatory drug nano preparation is characterized by comprising the following steps: cerium-mediated binding of serum albumin to non-steroidal anti-inflammatory drugs: dissolving serum albumin in water, adding trivalent cerium salt, reacting for 15min under magnetic stirring to synthesize a precursor, adding an alkaline solution into a reaction system, carrying out nucleation and curing reaction, adding a non-steroidal anti-inflammatory drug, and continuing to react for 15min, wherein the reaction system is in a nitrogen atmosphere.
3. The method of serum albumin-nsaid nanoformulation according to claims 1-2, characterized by: the serum albumin is bovine serum albumin, human serum albumin.
4. The method of serum albumin-nsaid nanoformulation according to claims 1-2, characterized by: the non-steroidal anti-inflammatory drug is an optional non-steroidal anti-inflammatory drug, such as sulindac, indomethacin, ibuprofen or aspirin.
5. The method of serum albumin-nsaid nanoformulation according to claims 1-2, characterized by: the composition of the serum albumin and the nonsteroidal anti-inflammatory drug and the cerium-mediated serum albumin and the nonsteroidal anti-inflammatory drug has a determined ratio, and the molar ratio of the serum albumin to the nonsteroidal anti-inflammatory drug is 0.1-1.5: 1-10; the molar ratio of the serum albumin, the trivalent cerium salt, the alkaline solution and the non-steroidal anti-inflammatory drug is 0.1-1.5: 1-10: 10-100: 0.1 to 1.5.
6. The method of claim 2, wherein the serum albumin-nsaid nanoformulation comprises: the trivalent cerium salt is cerous nitrate, cerous trichloride or cerous sulfate; the alkaline solution is sodium hydroxide, potassium hydroxide or ammonia water.
7. The method of claim 2, wherein the serum albumin-nsaid nanoformulation comprises: the nucleation curing reaction condition is that the reaction is carried out for 10min to 72h at the temperature of 4 to 80 ℃; the magnetic stirring is carried out at 500-1400 rpm/min.
8. The serum albumin and the nonsteroidal anti-inflammatory drug and the cerium-mediated serum albumin and the nonsteroidal anti-inflammatory drug which are prepared by the method according to any one of claims 1 to 7 are combined by a special chemical bond, and are a novel nano preparation, and the infrared spectrum of the nano preparation is different from that of a physical mixture of the serum albumin, the nonsteroidal anti-inflammatory drug, the serum albumin and the nonsteroidal anti-inflammatory drug.
9. The serum albumin-nsaid nano-preparation and the cerium-mediated serum albumin-nsaid nano-preparation according to claims 1 to 7 can be combined with optional one or more excipients or auxiliary materials to obtain corresponding pharmaceutical dosage forms or new drugs, and the pharmaceutical dosage forms or new drugs can be prepared into various other existing dosage forms according to the required purposes, directions and the like, and can be prepared by methods known in the art.
10. The serum albumin-NSAID nano preparation and the cerium-mediated serum albumin-NSAID nano preparation according to claims 1 to 7 have the function of treating diseases by using a non-steroidal anti-inflammatory drug or/and cerium nanoparticles, and the obtained serum albumin-NSAID nano preparation has a great promoting effect and an application prospect in the fields of drug targeted delivery, treatment of acute and chronic inflammatory diseases and regenerative medicine.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080187517A1 (en) * | 2004-04-06 | 2008-08-07 | Affibody Ab | Use of Serum Albumin Binding Peptides Conjugates for the Preparation of a Medicament |
| CN101543630A (en) * | 2009-04-20 | 2009-09-30 | 中国药科大学 | Preparation and application of amphiphilic albumin derivative and pharmaceutical composition thereof |
| CN104434808A (en) * | 2014-07-03 | 2015-03-25 | 石药集团中奇制药技术（石家庄）有限公司 | Therapeutic nanoparticles and preparation method thereof |
| CN105407718A (en) * | 2013-06-04 | 2016-03-16 | 维奥姆生物科学有限公司 | Coated particles and compositions comprising same |
| CN106270546A (en) * | 2016-08-12 | 2017-01-04 | 中国人民解放军第三军医大学军事预防医学院 | A kind of method of protein mediated synthetic modification cerium nano material |
| DE102017204850A1 (en) * | 2017-03-22 | 2018-09-27 | Michael Denck | HSA conjugate |
-
2021
- 2021-01-09 CN CN202110026825.0A patent/CN112704741A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080187517A1 (en) * | 2004-04-06 | 2008-08-07 | Affibody Ab | Use of Serum Albumin Binding Peptides Conjugates for the Preparation of a Medicament |
| CN101543630A (en) * | 2009-04-20 | 2009-09-30 | 中国药科大学 | Preparation and application of amphiphilic albumin derivative and pharmaceutical composition thereof |
| CN105407718A (en) * | 2013-06-04 | 2016-03-16 | 维奥姆生物科学有限公司 | Coated particles and compositions comprising same |
| CN104434808A (en) * | 2014-07-03 | 2015-03-25 | 石药集团中奇制药技术（石家庄）有限公司 | Therapeutic nanoparticles and preparation method thereof |
| CN106270546A (en) * | 2016-08-12 | 2017-01-04 | 中国人民解放军第三军医大学军事预防医学院 | A kind of method of protein mediated synthetic modification cerium nano material |
| DE102017204850A1 (en) * | 2017-03-22 | 2018-09-27 | Michael Denck | HSA conjugate |
Non-Patent Citations (6)
| Title |
|---|
| JING LIU，等: "Nucleus-Targeted Photosensitizer Nanoparticles for Photothermal and Photodynamic Therapy of Breast Carcinoma", 《INTERNATIONAL JOURNAL OF NANOMEDICINE》 * |
| QIAN CHEN，等: "Albumin Carriers for Cancer Theranostics: A Conventional Platform with New Promise", 《ADVANCED MATERIALS》 * |
| QIANWEN YU，等: "Targeting cancer-associated fibroblasts by dual-responsive lipid-albumin nanoparticles to enhance drug perfusion for pancreatic tumor therapy", 《JOURNAL OF CONTROLLED RELEASE》 * |
| YAN WEI,等: "Thermosensitive Liposomal Codelivery of HSA-Paclitaxel and HSA-Ellagic Acid Complexes for Enhanced Drug Perfusion and Efficacy Against Pancreatic Cancer", 《ACS APPLIED MATERIALS & INTERFACES 》 * |
| 吴艺舟，等: "白蛋白的功能化载体研究及临床应用", 《医学理论与实践》 * |
| 杨露勋，等: "铈纳米颗粒在辐射领域的研究进展", 《中国辐射卫生》 * |
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