CN109734901B - Polypeptide-based polyester ammonia-type nano particle and preparation and application thereof - Google Patents
Polypeptide-based polyester ammonia-type nano particle and preparation and application thereof Download PDFInfo
- Publication number
- CN109734901B CN109734901B CN201811573662.2A CN201811573662A CN109734901B CN 109734901 B CN109734901 B CN 109734901B CN 201811573662 A CN201811573662 A CN 201811573662A CN 109734901 B CN109734901 B CN 109734901B
- Authority
- CN
- China
- Prior art keywords
- polypeptide
- enzyme
- solution
- polyester ammonia
- polypeptidyl
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to a polypeptide-based polyester ammonia type nano particle, and preparation and application thereof, wherein the polypeptide-based polyester ammonia type nano particle is prepared by taking enzyme degradable polypeptide-based polyester ammonia as a raw material and degrading and assembling the raw material under the action of enzyme. The preparation method comprises the following steps: dissolving enzyme-degradable polypeptidyl polyesteramide in an organic solvent to obtain a polyesteramide solution with the mass fraction of 5-35%, removing the solvent to form a membrane, and degrading in an enzyme-containing PBS solution water bath shaking table with the concentration of 0.05-2.0 mg/mL to obtain the nanoparticles. The production process is safe, nontoxic and low in cost. The nano-particles have stable shape, uniform particle size distribution and excellent biocompatibility, can preload drugs, and are widely applied to the biomedical fields of wound antibiosis, bacterial biofilm inhibition, wound repair and the like.
Description
Technical Field
The invention belongs to the technical field of biomedical materials, and particularly relates to a polypeptide-based polyester ammonia-type nano particle, and preparation and application thereof.
Background
Degradable polymers are of interest for their wide range of applications, particularly in the biomedical field, such as controlled drug release, gene transfer, and tissue engineering. Biodegradable aliphatic polyesters and polycarbonates have become the most important synthetic biomaterials approved for biomedical device administration due to their good biocompatibility and the U.S. Food and Drug Administration (FDA). In practice, these classical biomedical polymers cannot meet the requirements of specific applications due to their high hydrophobicity, uncontrollable degradation rate, insufficient mechanical properties and other disadvantages.
Polyesteramines have been proposed as a new class of biomaterials comprising ester and amide linkages in their backbone for use in various biomedical applications. There are two classes of polyesteramines, one derived from non-amino acids, such as aliphatic diamines. Another class is derived from amino acids such as L-phenylalanine, L-leucine and L-lysine. The amino acid-based polyester has amido bonds and ester bonds in the molecular chain, so that the polymer has the characteristics of polyurethane and protein, namely, the biodegradation of enzyme-catalyzed surface erosion and the required mechanical, physical and biological compatibility are combined into a single entity. In the past decades, poly (ester amides) (PEAs) based on α -amino acids have been developed as a general class of biodegradable polymers with the advantageous properties of polyesters and polypeptides, such as enzymatic degradability and bioactivity.
Amino acid based polyester ammonia has a wide variety of material properties, good processability, excellent mechanical properties, also shows good biocompatibility and low inflammatory response, and may potentially enhance cell-substance interactions, is susceptible to enzymatic degradation by hydrolysis and hydrolysis, is biodegradable, their degradation products have a low toxicity source, amino acids can be absorbed by proteolytic enzymes and by the human body. However, amino acid based polyester ammonia has a single function, and modification in a molecular chain is required to provide the amino acid based polyester ammonia with multiple functions, so that the amino acid based polyester ammonia can be intelligently applied to specific fields.
The polypeptide is a vital active substance necessary for human body, affects many important physiological functions in the organism, has multiple functions, has protein and non-protein characteristics, and can be synthesized to imitate the characteristics of protein by designing amino acid types and sequences. Many active substances of human body exist in the form of peptide, and the polypeptide is a component and nutrient substance in human body, and can control growth, development and metabolism of human body, and at the same time, has the functions of preventing and curing diseases and regulating physiological function of human body. The short peptide has the characteristics of small molecular weight, unique action mechanism, no drug resistance, multiple functions and the like, is an ideal substitute, and can simulate more complex protein. The peptide chain can be applied to various biomedical fields such as treatment, biocatalysts, drug delivery, biosensing, intelligent biomaterial field and the like by designing the amino acid types and sequences and designing the physical, chemical, self-assembly, irritation and other properties of the peptide chain so as to simulate the properties of protein.
Wu Zhongshan university biomedical engineering institute professor, an article of "engineering-based poly (ester amide) nanoparticle platform" in Acta biomaterials journal, From structure-property correlation to nucleic acid delivery, through the steps of preparation of di-p-nitrophenyl ester of dicarboxylic acid monomer, synthesis of amino acid diester monomer, and preparation of polyesteramine, polyurethane and nucleic acid are formed into a complex, and nanoparticles are formed through electrostatic interaction for nucleic acid delivery. According to which 1) linker/spacer length (length effect and parity effect); 2) arginine in salt form; 3) a side chain; 4) the rigidity of the chain; 5) molecular Weight (MW), in vitro data further confirm that this nanoparticle exhibits excellent nucleic acid delivery properties. However, the polyester-ammonia material is only used as a matrix material, and needs to be modified in certain specific fields, for example, the polyester-ammonia material is prepared into a compound and is assembled into nanoparticles for application, and the problems of unstable structure, low drug loading rate, explosive release of drugs, incapability of long-term use and the like exist.
Disclosure of Invention
The invention aims to solve the technical problem of providing a polypeptide-based polyester ammonia type nano particle and preparation and application thereof, and overcomes the defects that the nano particle structure formed by the electrostatic action of the existing polyester ammonia material is unstable, the drug loading rate is low, the drug can be released suddenly, and the long-term use cannot be realized.
The invention relates to a polypeptide-based polyester ammonia type nano particle, which is prepared by taking enzyme degradable polypeptide-based polyester ammonia as a raw material and degrading and assembling the raw material under the action of enzyme.
The chemical structural formula of the enzyme-degradable multi-peptidyl polyester ammonia is as follows:
The enzyme is an enzyme for degrading amido bond or ester bond, and is selected from trypsin, alpha-chymotrypsin, lipase or phospholipase.
The invention also provides a preparation method of the nano-particles, which comprises the following steps:
dissolving enzyme-degradable polypeptidyl polyesteramide in an organic solvent to obtain a polyesteramide solution with the mass fraction of 5-35%, removing the solvent to form a membrane, and degrading in an enzyme-containing PBS solution water bath shaking table with the concentration of 0.05-2.0 mg/mL to obtain the nanoparticles.
The organic solvent is DMF, DMAC, THF or CHCl3。
The temperature of the water bath was 37 ℃.
The nano particles are frozen at the temperature of-19 ℃ and then freeze-dried and stored at the temperature of-50 ℃.
The invention further provides the application of the nanoparticles in drug loading, substances such as antibacterial agents, growth factors, S-nitrosylation glutathione and the like can be preloaded, the multifunctional property is endowed, the application of the polyester ammonia high molecular compound in the biomedical field is widened, and the nanoparticles can be used in the biomedical field of wound antibiosis, bacterial biofilm inhibition or wound repair.
The preparation method of the enzyme degradable type multi-peptidyl polyester ammonia comprises the following steps:
(1) dissolving polypeptide, anhydride and a catalyst in an organic solvent for reaction, adding the di-tert-butyl dicarbonate anhydride protected alcohol amine micromolecule, an activator and the catalyst for continuous reaction, then eluting with trifluoroacetic acid, filtering, carrying out rotary evaporation, precipitating, carrying out suction filtration and drying to obtain polypeptide-based diamine with amino groups at two ends, and storing at the temperature of below 0 ℃ in a sealed manner;
(2) dissolving p-nitrophenol in an organic solvent, adding a catalyst, dropwise adding diacyl chloride, stirring for reaction, continuously stirring at room temperature overnight, precipitating, filtering, washing, drying, recrystallizing to obtain p-dinitrobenzene active ester, sealing, drying and storing;
(3) and (2) dissolving the polypeptidyl diamine obtained in the step (1) and the p-dinitrobenzene active ester obtained in the step (2) in an organic solvent, adding a catalyst, carrying out solution polymerization, precipitating, filtering, purifying to obtain the polypeptidyl polyesteramide, sealing, drying and storing.
The polypeptide in the step (1) is synthesized by a solid phase synthesis method.
The number of carbon atoms of the acid anhydride in the step (1) is 2-12, and the acid anhydride is selected from succinic anhydride or phthalic anhydride.
The organic solvent in the step (1) is DMF, DMAC, THF or CHCl3。
The activating agent in the step (1) is an activating agent for activating carboxyl, and is selected from DCC, EDC or NHS.
The catalyst in the step (1) is a basic catalyst selected from DMAP, DIEA and ET3N and one or more of pyridine.
The number of carbon atoms of the di-tert-butyl dicarbonate anhydride-protected alcohol amine micromolecules in the step (1) is 2-8, one end of a carbon chain is an amino group, and the other end of the carbon chain is an alcoholic hydroxyl group, and the alcohol amine is selected from ethanolamine, isopropanolamine or isobutanol amine.
The structural formula of the polypeptide-based diamine prepared in the step (1) is as follows:
The technological parameters of the reaction in the step (1) are as follows: the reaction temperature is normal temperature, and the reaction time is 1-5 h.
The technological parameters of the continuous reaction in the step (1) are as follows: the reaction temperature is-10 to 0 ℃, and the reaction time is 18 to 24 hours.
The elution process conditions in the step (1) are as follows: eluting with 0.25-5.0 wt% trifluoroacetic acid for 3-5 times. The purpose is to remove the di-tert-butyl dicarbonate anhydride protecting group and to cleave the resin.
The process conditions of the precipitation in the step (1) are as follows: the product was precipitated with cold ether.
The organic solvent in the step (2) is acetone, DMF, DMAC, THF and CHCl3One or more of them.
The catalyst in the step (2) is a basic catalyst selected from DMAP, DIEA and ET3N and one or more of pyridine.
The diacid chloride in the step (2) is aliphatic diacid chloride, and is selected from succinyl chloride, glutaryl chloride, adipoyl chloride or sebacoyl chloride.
The technological parameters of the stirring reaction in the step (2) are as follows: the stirring is mechanical stirring, the stirring reaction temperature is-90 to-30 ℃, and the stirring reaction time is 1 to 3 hours.
The process conditions of the precipitation in the step (2) are as follows: the product was precipitated with distilled water.
The process conditions of recrystallization in the step (2) are as follows: and recrystallizing with ethyl acetate for 3 times.
The organic solvent in the step (3) is DMF, DMAC or DMSO.
The catalyst in the step (3) is a basic catalyst selected from DMAP, DIEA and ET3N and one or more of pyridine.
The process parameters of the solution polymerization in the step (3) are as follows: the polymerization temperature is 60-100 ℃, and the reaction time is 6-24 h.
The process conditions of the precipitation in the step (3) are as follows: the product was precipitated with cold ethyl acetate.
The polypeptide-based polyester ammonia is a relatively new-generation synthetic biodegradable biomaterial, and due to the existence of polypeptide micromolecules, the polypeptide-based polyester ammonia has protein and non-protein characteristics and can simulate certain characteristics of proteins; most of the molecular chains are ester and amide groups, which provide chemically functionalized reaction sites. In addition, the most unique aspect is the biological characteristic of the polypeptide, because the polypeptide is 2-8 peptide consisting of hydrophilic and hydrophobic amino acids, the polypeptide is synthesized to imitate the characteristic energy of protein by designing the type and the sequence of the amino acid, and the polypeptide is micromolecule amino acid after being degraded, and is nontoxic and harmless to the environment and the human body, so the polymer has good biocompatibility and can not cause inflammatory reaction due to foreign matters; a large number of amido bonds and ester bonds in the polymer can be used as enzyme degradation sites and can be degraded by a plurality of enzymes, the degradation rate of the ester bonds and the amido bonds can be degraded by one or more enzymes, the polyester ammonia polymer is degraded in an enzyme solution, hydrophilic segments and hydrophobic segments in a molecular chain are assembled to finally obtain nano particles, the nano particles with different particle sizes are obtained by adjusting the type, concentration and degradation time of the enzyme, the nano particles are degraded into spheres, and the application of the nano particles is widened.
The nano-particles related by the invention are polypeptide-based nano-particles initiated by enzyme, the synthesized polymer is degraded by the enzyme, the polymer coats the drug in the degradation process, the drug is firmly locked in the nano-particles, and meanwhile, the nano-structure can change along with the change of concentration, so that the stability is controllable, the drug loading rate is obviously improved, and the nano-particles can be used for a long time. In addition, the nano particles can transmit various medicaments, thereby greatly widening the application field of the polyester-ammonia material.
Advantageous effects
(1) The production process of the invention is safe, nontoxic and low in cost.
(2) The polypeptide-based polyester ammonia-type nano particle is in a stable spherical shape, has a stable structure and is uniform in particle size distribution.
(3) The polypeptide-based polyester ammonia type nano particle can preload medicines, comprises S-nitrosoglutathione, antibacterial agents, growth factors and other substances, has excellent biocompatibility, and can be widely applied to the biomedical fields of wound antibiosis, biomembrane inhibition, wound repair and the like.
Drawings
FIG. 1 is a scanning electron microscope image of the amino-type nano-particles of the polypeptidyl polyester prepared in example 1 of the invention.
FIG. 2 is a particle size distribution diagram of the amino-type nanoparticles of the polypeptidyl polyester prepared in example 1 of the present invention.
FIG. 3 is a scanning electron microscope image of Escherichia coli treated by the antibiotic-free polypeptidyl polyester ammonia-type nanoparticles prepared in example 3 of the present invention.
FIG. 4 is a scanning electron microscope image of Escherichia coli treated by the antibacterial agent-containing polypeptidyl polyester ammonia-type nanoparticles prepared in example 3 of the present invention.
Detailed Description
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, it should be understood that various changes or modifications of the present invention may be made by those skilled in the art after reading the teaching of the present invention, and such equivalents may fall within the scope of the present invention as defined in the appended claims.
Example 1
1. Preparation of polypeptidyl diamines
(1) Preparation of tripeptides:
using standard FMOC Solid Phase Peptide Synthesis (SPPS) techniques, the reaction involved the following ratios of materials: 2g of 2-chlorotrityl chloride resin, 1.6mmol of FMOC-Lys (Boc) -OH 3.78g, 2.48g of 6.4mmol of FMOC-Phe-OH, 2.26g of 6.4mmol of FMOC-Leu-OH, 2.42g of 6.4mmol of HBTU, 0.87g of 6.4mmol of HOBt, 3ml of 6.4mmol of DIEA and 5ml of piperidine. The method comprises the following steps:
adding the resin into a polypeptide synthesis device, adding dry DMF, soaking for half an hour to fully swell the resin, and finally discharging the solvent DMF.
Dissolving amino acid with DMF, transferring the solution into the polypeptide synthesis device containing the treated resin, adding catalyst DIEA, reacting at room temperature for 1.5h to fully fix the resin, and washing the resin with DMF.
piperidine/DMF solution was added to the resin from the previous step for half an hour, deprotected, the resin washed with DMF and checked for complete protection with ninhydrin.
Dissolving amino acid, HBTU and HOBt in DMF, transferring the solution into the polypeptide synthesis device containing the treated resin, adding a catalyst DIEA, reacting at room temperature for 1.5h, washing the resin with DMF, and detecting whether the amino group is completely reacted with ninhydrin, wherein if the amino group is colorless, the condensation reaction can be carried out for the next step; if blue color is developed, the reaction solution is condensed to colorless, and then the next operation can be carried out.
The above procedure was repeated until colorless as checked with ninhydrin, indicating complete amino reaction to give the tripeptide.
(2) Preparation of di-tert-butyl dicarbonate anhydride-protected ethanolamine:
ethanolamine (10.0ml, 165mmol) in anhydrous CH at-10 deg.C2Cl2To a solution in (500mL) was added triethylamine (24.5mL, 250mmol) followed by di-tert-butyl dicarbonate anhydride (36g, 165 mmol). The solution was stirred at 25 ℃ for 20 hours and then saturated NHCl4The solution (100ml) was quenched. The aqueous layer was extracted with ethyl acetate (3X 200 ml). The combined organic layers were then washed with brine, over MgSO4Drying and concentration under reduced pressure gave di-tert-butyl dicarbonate anhydride-protected ethanolamine as a colorless oil.
(3) Reaction of tripeptide with succinic anhydride, di-tert-butyl dicarbonate anhydride protected ethanolamine:
succinic anhydride (0.64g, 6.4mmol), DIEA (3ml), and ET were added to the tripeptide DMF solution obtained above3N (0.65g, 6.4mmol) was reacted at room temperature for 5 hours. Then, di-tert-butyl dicarbonate anhydride-protected ethanolamine (1.03g, 4mmol), DCC (1.32g, 6.4mmol), DMAP (0.78g, 6.4mmol) was dissolved in DMF and added at 0 ℃ to react for 24 hours. After draining the DMF solution, cleavage of the peptide and removal of the protected di-tert-butyl dicarbonate anhydride groups were carried out using trifluoroacetic acid at a concentration of 5% by weight. After shaking at room temperature for 2 hours, the mixture was collected. The combined solution was concentrated to a viscous solution by rotary evaporation. Adding cold ether to precipitate the product, dissolving the precipitate in distilled water, and freeze-drying under vacuum to obtain white product, i.e. polypeptidyl diamine, and storing at a temperature below 0 ℃ in a sealed manner.
2. Preparation of p-dinitrobenzene active ester
A solution of triethylamine (0.0804mol) and p-nitrophenol (0.0804mol) in 100ml of acetone was kept at-78 ℃ at room temperature with dry ice and acetone. Succinyl chloride (0.04mol) in 80ml acetone was then added dropwise to the cooled solution, stirred at-78 ℃ for 2 hours, then stirred at room temperature overnight. Thereafter, the mixture was poured into 1000ml of distilled water to precipitate the product, which was filtered, washed thoroughly with distilled water, dried under vacuum at 50 ℃ and finally purified by recrystallization from ethyl acetate for 3 times to give needle-like off-white solid p-dinitrobenzene active ester, which was stored sealed and dried.
3. Preparation of enzyme degradable polypeptide-based polyester ammonia
The polypeptidyl diamine (1.0mmol) and the p-dinitrobenzene active ester (1mmol) were dissolved in 1.5ml of anhydrous DMAC and the solution was heated to 60 ℃ with stirring until the monomers were completely dissolved. Will ET3N (2.2mmol) was added dropwise to the solution, and the reaction was held at 80 ℃ for 16 hours to effect polymerization. Precipitating the obtained solution with cold ethyl acetate, filtering, extracting with ethyl acetate in a Soxhlet extractor for 48 hours, finally drying at 50 ℃ in vacuum to obtain white solid polypeptide-based polyesteramide, sealing, drying and storing.
4. Degradation of enzymatically degradable polypeptidyl polyesteramines
Dissolving the polypeptidyl polyesteramide in DMF to form a polymer solution with the mass fraction of 30%, removing the solvent to form a film, placing the obtained film in a PBS solution with the lipase concentration of 0.15mg/ml, degrading the polymer in a water bath shaker at 37 ℃ at a certain shaking speed to obtain nanoparticles, freezing at-19 ℃, and freeze-drying and storing at-50 ℃.
As shown in a scanning electron microscope picture of the polypeptide-based polyester ammonia-type nano particles prepared by the embodiment, as shown in figure 1, the particles are spherical, have no obvious agglomeration phenomenon and have a stable structure.
As shown in FIG. 2, the particle size distribution diagram of the polypeptide-based polyester ammonia-type nanoparticles prepared in this example shows that the particle size of the nanoparticles is maintained at about 200nm and the particle size distribution is uniform.
Example 2
1. Preparation of polypeptidyl diamines
(1) Preparation of tetrapeptide:
using standard FMOC Solid Phase Peptide Synthesis (SPPS) techniques, the reaction involved the following ratios of materials: 2g of 2-chlorotrityl chloride resin, 1.6mmol of FMOC-Lys (Boc) -OH 3.78g, 2.48g of 6.4mmol of FMOC-Phe-OH, 2.26g of 6.4mmol of FMOC-Leu-OH, 2.42g of 6.4mmol of HBTU, 0.87g of 6.4mmol of HOBt, 3ml of 6.4mmol of DIEA and 5ml of piperidine. The method comprises the following steps:
adding the resin into a polypeptide synthesis device, adding dry DMF, soaking for half an hour to fully swell the resin, and finally discharging the solvent DMF.
Dissolving amino acid with DMF, transferring the solution into the polypeptide synthesis device containing the treated resin, adding catalyst DIEA, reacting at room temperature for 1.5h to fully fix the resin, and washing the resin with DMF.
piperidine/DMF solution was added to the resin from the previous step for half an hour, deprotected, the resin washed with DMF and checked for complete protection with ninhydrin.
Dissolving amino acid, HBTU and HOBt in DMF, transferring the solution into the polypeptide synthesis device containing the treated resin, adding a catalyst DIEA, reacting at room temperature for 1.5h, washing the resin with DMF, and detecting whether the amino group is completely reacted with ninhydrin, wherein if the amino group is colorless, the condensation reaction can be carried out for the next step; if blue color is developed, the reaction solution is condensed to colorless, and then the next operation can be carried out.
The above steps were repeated until colorless as checked by ninhydrin, indicating complete amino reaction to give the tetrapeptide.
(2) Preparation of di-tert-butyl dicarbonate anhydride-protected ethanolamine:
ethanolamine (10.0ml, 165mmol) in anhydrous CH at-10 deg.C2Cl2To a solution in (500mL) was added triethylamine (24.5mL, 250mmol) followed by di-tert-butyl dicarbonate anhydride (36g, 165 mmol). The solution was stirred at 25 ℃ for 20 hours and then saturated NHCl4The solution (100ml) was quenched. The aqueous layer was extracted with ethyl acetate (3X 200 ml). The combined organic layers were then washed with brine, over MgSO4Drying and concentration under reduced pressure gave di-tert-butyl dicarbonate anhydride-protected ethanolamine as a colorless oil.
(3) Reaction of tetrapeptide with succinic anhydride, di-tert-butyl dicarbonate anhydride protected ethanolamine:
to the solution of tetrapeptide in DMF obtained above was added succinic anhydride (0.64g, 6.4mmol), DIEA (3ml), ET3N (0.65g, 6.4mmol) was reacted at room temperature for 5 hours. Then, di-tert-butyl dicarbonate anhydride-protected ethanolamine (1.03g, 4mmol), DCC (1.32g, 6.4mmol), DMAP (0.78g, 6.4mmol) was dissolved in DMF and added at 0 ℃ to react for 24 hours. After draining the DMF solution, cleavage of the peptide and removal of the protected di-tert-butyl dicarbonate anhydride groups were carried out using trifluoroacetic acid at a concentration of 5% by weight. After shaking at room temperature for 2 hours, the mixture was collected. The combined solution was concentrated to a viscous solution by rotary evaporation. Adding cold ether to precipitate the product, dissolving the precipitate in distilled water, and freeze-drying under vacuum to obtain white product, i.e. polypeptidyl diamine, and storing at a temperature below 0 ℃ in a sealed manner.
2. Preparation of p-dinitrobenzene active ester
A solution of triethylamine (0.0804mol) and p-nitrophenol (0.0804mol) in 100ml of acetone was kept at-78 ℃ at room temperature with dry ice and acetone. Succinyl chloride (0.04mol) in 80ml acetone was then added dropwise to the cooled solution, stirred at-78 ℃ for 2 hours, then stirred at room temperature overnight. Thereafter, the mixture was poured into 1000ml of distilled water to precipitate the product, which was filtered, washed thoroughly with distilled water, dried under vacuum at 50 ℃ and finally purified by recrystallization from ethyl acetate for 3 times to give needle-like off-white solid p-dinitrobenzene active ester, which was stored sealed and dried.
3. Preparation of enzyme degradable polypeptide-based polyester ammonia
The polypeptidyl diamine (1.0mmol) and the p-dinitrobenzene active ester (1mmol) were dissolved in 1.5ml of anhydrous DMAC and the solution was heated to 60 ℃ with stirring until the monomers were completely dissolved. Will ET3N (2.2mmol) was added dropwise to the solution, and the reaction was held at 80 ℃ for 16 hours to effect polymerization. Precipitating the obtained solution with cold ethyl acetate, filtering, extracting with ethyl acetate in a Soxhlet extractor for 48 hours, finally drying at 50 ℃ in vacuum to obtain white solid polypeptide-based polyesteramide, sealing, drying and storing.
4. Degradation of enzymatically degradable polypeptidyl polyesteramines
Dissolving the polypeptidyl polyesteramide in DMF to form a polymer solution with the mass fraction of 30%, removing the solvent to form a film, placing the obtained film in a PBS solution with the trypsin concentration of 1.0mg/ml, wherein the PBS solution contains the antibiotic drug levofloxacin with the concentration of 1.5ml, and degrading the polymer in a water bath shaker at 37 ℃ at a certain shaking speed to obtain drug-loaded nanoparticles, freezing at-19 ℃ and freeze-drying and storing at-50 ℃.
The polypeptide-based polyester ammonia-type nanoparticles prepared by the embodiment are spherical, have no obvious agglomeration phenomenon and have a stable structure; the particle size distribution is uniform, and the particle size is kept about 200-300 nm.
The drug loading performance of the antibiotic-preloaded polypeptide-based polyester ammonia type nanoparticles prepared by the embodiment can be characterized by the drug loading rate, wherein the drug loading rate is drug loading/(drug loading + carrier mass), and the drug loading rate can be detected to be 40% by combining with infrared, simple, nuclear magnetic and ultraviolet molecular methods. The nano particles act on escherichia coli, the morphology of bacteria is observed through a scanning electron microscope image, the cell wall of the escherichia coli is damaged, the structure of the escherichia coli is damaged, the shape is flat, the escherichia coli is dead, and the antibacterial effect of the nano particles can be visually seen.
Example 3
1. Preparation of polypeptidyl diamines
(1) Preparation of tetrapeptide: the tetrapeptide was prepared by the method of preparation of the tetrapeptide in example 2.
(2) Preparation of di-tert-butyl dicarbonate anhydride-protected ethanolamine: ethanolamine (10.0ml, 165mmol) in anhydrous CH at-10 deg.C2Cl2To a solution in (500mL) was added triethylamine (24.5mL, 250mmol) followed by di-tert-butyl dicarbonate anhydride (36g, 165 mmol). The solution was stirred at 25 ℃ for 20 hours and then saturated NHCl4The solution (100ml) was quenched. The aqueous layer was extracted with ethyl acetate (3X 200 ml). The combined organic layers were then washed with brine, over MgSO4Drying and concentration under reduced pressure gave di-tert-butyl dicarbonate anhydride-protected ethanolamine as a colorless oil.
(3) Reaction of tetrapeptide with phthalic anhydride, di-tert-butyl dicarbonate anhydride protected ethanolamine: to the solution of tetrapeptide in DMF obtained above was added phthalic anhydride (0.89g, 6.4mmol), DIEA (3ml), ET3N (0.65g, 6.4mmol) was reacted at room temperature for 5 hours. Then, di-tert-butyl dicarbonate anhydride-protected ethanolamine (1.03g, 4mmol), DCC (1.32g, 6.4mmol), DMAP (0.78g, 6.4mmol) was dissolved in DMF and added at 0 ℃ to react for 24 hours. After draining the DMF solution, cleavage of the peptide and removal of the protected di-tert-butyl dicarbonate anhydride groups were carried out using trifluoroacetic acid at a concentration of 5% by weight. After shaking at room temperature for 2 hours, the mixture was collected. The combined solution was concentrated to a viscous solution by rotary evaporation. Adding cold ether to precipitate the product, dissolving the precipitate in distilled water, and freeze-drying under vacuum to obtain white product, i.e. polypeptidyl diamine, and storing at a temperature below 0 ℃ in a sealed manner.
2. Preparation of p-dinitrobenzene active ester
A solution of triethylamine (0.0804mol) and p-nitrophenol (0.0804mol) in 100ml of acetone was kept at-78 ℃ at room temperature with dry ice and acetone. Glutaryl chloride (0.04mol) in 80ml of acetone was then added dropwise to the cooled solution, stirred at-78 ℃ for 2 hours and then at room temperature overnight. Thereafter, the mixture was poured into 1000ml of distilled water to precipitate the product, which was filtered, washed thoroughly with distilled water, dried under vacuum at 50 ℃ and finally purified by recrystallization from ethyl acetate for 3 times to give needle-like off-white solid p-dinitrobenzene active ester, which was stored sealed and dried.
3. Preparation of enzyme degradable polypeptide-based polyester ammonia
The polypeptidyl diamine (1.0mmol) and the p-dinitrobenzene active ester (1mmol) were dissolved in 1.5ml of anhydrous DMAC and the solution was heated to 60 ℃ with stirring until the monomers were completely dissolved. Will ET3N (2.2mmol) was added dropwise to the solution, and the reaction was held at 80 ℃ for 16 hours to effect polymerization. Precipitating the obtained solution with cold ethyl acetate, filtering, extracting with ethyl acetate in a Soxhlet extractor for 48 hours, finally drying at 50 ℃ in vacuum to obtain white solid polypeptide-based polyesteramide, sealing, drying and storing.
4. Degradation of enzymatically degradable polypeptidyl polyesteramines
Dissolving the polypeptidyl polyesteramide in DMF to form a polymer solution with the mass fraction of 30%, removing the solvent to form a film, placing the obtained film in a PBS solution with the trypsin concentration of 1.5mg/ml, degrading the polymer in a water bath shaker at 37 ℃ at a certain shaking speed to obtain the nano-particles without the antibacterial agent, freezing at 19 ℃ below zero, and freeze-drying and storing at 50 ℃ below zero.
Dissolving the polypeptidyl polyesteramide in DMF to form a polymer solution with the mass fraction of 30%, removing the solvent to form a film, placing the obtained film in a PBS solution with the trypsin concentration of 1.5mg/ml, wherein the PBS solution contains the antibiotic drug levofloxacin with the concentration of 1.2ml, and degrading the polymer in a water bath shaker at 37 ℃ at a certain shaking speed to obtain the nano-particles containing the antibacterial agent, freezing at-19 ℃ and freeze-drying and storing at-50 ℃.
The polypeptide-based polyester ammonia-type nanoparticles prepared by the embodiment are spherical, have no obvious agglomeration phenomenon and have a stable structure; the particle size distribution is uniform, and the particle size is kept about 200-300 nm.
The polypeptide-based polyester ammonia-type nanoparticles prepared in this example were used for treating escherichia coli, and as shown in the scanning electron microscope image of fig. 3, it was found that under such conditions, the cell walls of escherichia coli were continuous and not damaged, the structure thereof was complete, and a good rod-like morphology could be maintained.
The drug loading performance of the polypeptide-based polyester ammonia-type nanoparticle preloaded with the antibacterial agent prepared in the embodiment can be characterized by the drug loading rate, wherein the drug loading rate is loaded with drugs/(loaded drugs + carrier mass), in addition, the drug loading rate is 60% by detection in combination with infrared, simple, nuclear magnetic and ultraviolet molecular methods, and the like, and the drug loading rate is used for treating escherichia coli, as shown in a Scanning Electron Microscope (SEM) picture of 4, the cell wall of the escherichia coli is known to be damaged, the structure is damaged, the shape is flat, and the escherichia coli is known to die, which indicates that the nanoparticle has an antibacterial effect.
Example 4
1. Preparation of polypeptidyl diamines
(1) Preparation of tetrapeptide: the tetrapeptide was prepared by the method of preparation of the tetrapeptide in example 2.
(2) Preparation of di-tert-butyl dicarbonate anhydride-protected isopropanolamine: isopropanolamine (12.5ml, 165mmol) in anhydrous CH at-10 deg.C2Cl2To a solution in (500mL) was added triethylamine (24.5mL, 250mmol) followed by di-tert-butyl dicarbonate anhydride (36g, 165 mmol). The solution was stirred at 25 ℃ for 20 hours and then saturated NHCl4The solution (100ml) was quenched. The aqueous layer was extracted with ethyl acetate (3X 200 ml). The combined organic layers were then washed with brine, over MgSO4Drying and concentration under reduced pressure gave di-tert-butyl dicarbonate anhydride-protected isopropanolamine as a colorless oil.
(3) Reaction of tetrapeptide with succinic anhydride, di-tert-butyl dicarbonate anhydride protected isopropanolamine: to the solution of tetrapeptide in DMF obtained above was added succinic anhydride (0.64g, 6.4mmol), DIEA (3ml), ET3N (0.65g, 6.4mmol) was reacted at room temperature for 5 hours. Di-tert-butyl dicarbonate anhydride-protected isopropanolamine (0.75g, 4mmol), DCC (1.32g, 6.4mmol), DMAP (0.78g, 6.4mmol) was then dissolved in DMF and added at 0 ℃ for reaction for 24 hours. After the DMF solution was drained, cleavage of the peptide and removal of the protected di-tert-butyl dicarbonate anhydride group were carried out using trifluoroacetic acid at a concentration of 4.5% by weight. In the roomAfter 2 hours of shaking at room temperature, the mixture was collected. The combined solution was concentrated to a viscous solution by rotary evaporation. Adding cold ether to precipitate the product, dissolving the precipitate in distilled water, and freeze-drying under vacuum to obtain white product, i.e. polypeptidyl diamine, and storing at a temperature below 0 ℃ in a sealed manner.
2. Preparation of p-dinitrobenzene active ester
A solution of triethylamine (0.0804mol) and p-nitrophenol (0.0804mol) in 100ml of acetone was kept at-78 ℃ at room temperature with dry ice and acetone. Succinyl chloride (0.04mol) in 80ml acetone was then added dropwise to the cooled solution, stirred at-78 ℃ for 2 hours, then stirred at room temperature overnight. Thereafter, the mixture was poured into 1000ml of distilled water to precipitate the product, which was filtered, washed thoroughly with distilled water, dried under vacuum at 50 ℃ and finally purified by recrystallization from ethyl acetate for 3 times to give needle-like off-white solid p-dinitrobenzene active ester, which was stored sealed and dried.
3. Preparation of enzyme degradable polypeptide-based polyester ammonia
The polypeptidyl diamine (1.0mmol) and the p-dinitrobenzene active ester (1mmol) were dissolved in 1.5ml of anhydrous DMAC and the solution was heated to 60 ℃ with stirring until the monomers were completely dissolved. Will ET3N (2.2mmol) was added dropwise to the solution, and the reaction was held at 80 ℃ for 16 hours to effect polymerization. Precipitating the obtained solution with cold ethyl acetate, filtering, extracting with ethyl acetate in a Soxhlet extractor for 48 hours, finally drying at 50 ℃ in vacuum to obtain white solid polypeptide-based polyesteramide, sealing, drying and storing.
4. Degradation of enzymatically degradable polypeptidyl polyesteramines
Dissolving the polypeptidyl polyesteramide in DMF to form a polymer solution with the mass fraction of 30%, removing the solvent to form a film, placing the obtained film in a PBS solution with the trypsin concentration of 2mg/ml, wherein the PBS solution contains 0.5mg/ml of fibroblast growth factor (VEGF), and degrading the polymer in a water bath shaker at 37 ℃ at a certain shaking speed to obtain drug-loaded nanoparticles, freezing at-19 ℃ and freeze-drying and storing at-50 ℃.
The polypeptide-based polyester ammonia-type nanoparticles prepared by the embodiment are spherical, have no obvious agglomeration phenomenon and have a stable structure; the particle size distribution is uniform, and the particle size is kept about 200-400 nm.
The drug loading performance of the growth factor preloaded polypeptide-based polyester ammonia type nanoparticles prepared by the embodiment can be characterized by the drug loading rate, wherein the drug loading rate is drug loading/(drug loading + carrier mass), in addition, the drug loading rate is 55% by detection through combining infrared, simple, nuclear magnetic and ultraviolet molecular methods, and the like, and the drug loaded nanoparticles and human fibroblasts are placed in a container containing 5% of CO2And culturing in a constant-temperature incubator at 37 ℃ for a certain time, observing the shape and the number of the cells under an electron microscope, and thus, the number of the cells is obviously increased, so that the effect of promoting cell proliferation is verified.
Example 5
1. Preparation of polypeptidyl diamines
(1) Preparation of tripeptides: the tetrapeptide was prepared by the method for preparing the tripeptide of example 1.
(2) Preparation of di-tert-butyl dicarbonate anhydride-protected ethanolamine:
ethanolamine (10.0ml, 165mmol) in anhydrous CH at-10 deg.C2Cl2To a solution in (500mL) was added triethylamine (24.5mL, 250mmol) followed by di-tert-butyl dicarbonate anhydride (36g, 165 mmol). The solution was stirred at 25 ℃ for 20 hours and then saturated NHCl4The solution (100ml) was quenched. The aqueous layer was extracted with ethyl acetate (3X 200 ml). The combined organic layers were then washed with brine, over MgSO4Drying and concentration under reduced pressure gave di-tert-butyl dicarbonate anhydride-protected ethanolamine as a colorless oil.
(3) Reaction of tripeptide with succinic anhydride, di-tert-butyl dicarbonate anhydride protected ethanolamine:
succinic anhydride (0.64g, 6.4mmol), DIEA (3ml), and ET were added to the tripeptide DMF solution obtained above3N (0.65g, 6.4mmol) was reacted at room temperature for 5 hours. Then, di-tert-butyl dicarbonate anhydride-protected ethanolamine (1.03g, 4mmol), DCC (1.32g, 6.4mmol), DMAP (0.78g, 6.4mmol) was dissolved in DMF and added at 0 ℃ to react for 24 hours. After the DMF solution was taken off, the concentration was 5wt% trifluoroacetic acid, cleavage of the peptide and removal of the protected di-tert-butyl dicarbonate anhydride group. After shaking at room temperature for 2 hours, the mixture was collected. The combined solution was concentrated to a viscous solution by rotary evaporation. Adding cold ether to precipitate the product, dissolving the precipitate in distilled water, and freeze-drying under vacuum to obtain white product, i.e. polypeptidyl diamine, and storing at a temperature below 0 ℃ in a sealed manner.
2. Preparation of p-dinitrobenzene active ester
A solution of triethylamine (0.0804mol) and p-nitrophenol (0.0804mol) in 100ml of acetone was kept at-78 ℃ at room temperature with dry ice and acetone. Glutaryl chloride (0.04mol) in 80ml of acetone was then added dropwise to the cooled solution, stirred at-78 ℃ for 2 hours and then at room temperature overnight. Thereafter, the mixture was poured into 1000ml of distilled water to precipitate the product, which was filtered, washed thoroughly with distilled water, dried under vacuum at 50 ℃ and finally purified by recrystallization from ethyl acetate for 3 times to give needle-like off-white solid p-dinitrobenzene active ester, which was stored sealed and dried.
3. Preparation of enzyme degradable polypeptide-based polyester ammonia
The polypeptidyl diamine (1.0mmol) and the p-dinitrobenzene active ester (1mmol) were dissolved in 1.5ml of anhydrous DMAC and the solution was heated to 60 ℃ with stirring until the monomers were completely dissolved. Will ET3N (2.2mmol) was added dropwise to the solution, and the reaction was held at 80 ℃ for 16 hours to effect polymerization. Precipitating the obtained solution with cold ethyl acetate, filtering, extracting with ethyl acetate in a Soxhlet extractor for 48 hours, finally drying at 50 ℃ in vacuum to obtain white solid polypeptide-based polyesteramide, sealing, drying and storing.
4. Degradation of enzymatically degradable polypeptidyl polyesteramines
Dissolving the polypeptidyl polyesteramide in DMF to form a polymer solution with the mass fraction of 30%, removing the solvent to form a film, placing the obtained film in a PBS solution with the chymotrypsin concentration of 0.5mg/ml, wherein the PBS solution contains 0.3mg/ml of fibroblast growth factor (VEGF), and degrading the polymer in a water bath shaker at 37 ℃ at a certain shaking speed to obtain drug-loaded nanoparticles, freezing at-19 ℃ and freeze-drying and storing at-50 ℃.
The polypeptide-based polyester ammonia-type nanoparticles prepared by the embodiment are spherical, have no obvious agglomeration phenomenon and have a stable structure; the particle size distribution is uniform, and the particle size is kept about 200-400 nm.
The drug loading performance of the growth factor preloaded polypeptide-based polyester ammonia type nanoparticles prepared by the embodiment can be characterized by the drug loading rate, wherein the drug loading rate is drug loading/(drug loading + carrier mass), in addition, the drug loading rate is 55% by detection through combining infrared, simple, nuclear magnetic and ultraviolet molecular methods, and the like, and the drug loaded nanoparticles and human fibroblasts are placed in a container containing 5% of CO2And culturing in a constant-temperature incubator at 37 ℃ for a certain time, observing the shape and the number of the cells under an electron microscope, and thus, the number of the cells is obviously increased, so that the effect of promoting cell proliferation is verified.
Example 6
1. Preparation of polypeptidyl diamines
(1) Preparation of pentapeptide:
using standard FMOC Solid Phase Peptide Synthesis (SPPS) techniques, the reaction involved the following ratios of materials: 2g of 2-chlorotrityl chloride resin, 1.6mmol of FMOC-Lys (Boc) -OH 3.78g, 2.48g of 6.4mmol of FMOC-Phe-OH, 2.26g of 6.4mmol of FMOC-Leu-OH, 2.42g of 6.4mmol of HBTU, 0.87g of 6.4mmol of HOBt, 3ml of 6.4mmol of DIEA and 5ml of piperidine. The method comprises the following steps:
adding the resin into a polypeptide synthesis device, adding dry DMF, soaking for half an hour to fully swell the resin, and finally discharging the solvent DMF.
Dissolving amino acid with DMF, transferring the solution into the polypeptide synthesis device containing the treated resin, adding catalyst DIEA, reacting at room temperature for 1.5h to fully fix the resin, and washing the resin with DMF.
piperidine/DMF solution was added to the resin from the previous step for half an hour, deprotected, the resin washed with DMF and checked for complete protection with ninhydrin.
Dissolving amino acid, HBTU and HOBt in DMF, transferring the solution into the polypeptide synthesis device containing the treated resin, adding a catalyst DIEA, reacting at room temperature for 1.5h, washing the resin with DMF, and detecting whether the amino group is completely reacted with ninhydrin, wherein if the amino group is colorless, the condensation reaction can be carried out for the next step; if blue color is developed, the reaction solution is condensed to colorless, and then the next operation can be carried out.
The above steps were repeated until colorless as checked with ninhydrin, indicating complete amino reaction to give the pentapeptide.
(2) Preparation of di-tert-butyl dicarbonate anhydride-protected ethanolamine:
ethanolamine (10.0ml, 165mmol) in anhydrous CH at-10 deg.C2Cl2To a solution in (500mL) was added triethylamine (24.5mL, 250mmol) followed by di-tert-butyl dicarbonate anhydride (36g, 165 mmol). The solution was stirred at 25 ℃ for 20 hours and then saturated NHCl4The solution (100ml) was quenched. The aqueous layer was extracted with ethyl acetate (3X 200 ml). The combined organic layers were then washed with brine, over MgSO4Drying and concentration under reduced pressure gave di-tert-butyl dicarbonate anhydride-protected ethanolamine as a colorless oil.
(3) Reaction of pentapeptide with succinic anhydride, di-tert-butyl dicarbonate anhydride protected ethanolamine:
to the pentapeptide DMF solution obtained above was added succinic anhydride (0.64g, 6.4mmol), DIEA (3ml), ET3N (0.65g, 6.4mmol) was reacted at room temperature for 5 hours. Then, di-tert-butyl dicarbonate anhydride-protected ethanolamine (1.03g, 4mmol), DCC (1.32g, 6.4mmol), DMAP (0.78g, 6.4mmol) was dissolved in DMF and added at 0 ℃ to react for 24 hours. After draining the DMF solution, cleavage of the peptide and removal of the protected di-tert-butyl dicarbonate anhydride groups were carried out using trifluoroacetic acid at a concentration of 5% by weight. After shaking at room temperature for 2 hours, the mixture was collected. The combined solution was concentrated to a viscous solution by rotary evaporation. Adding cold ether to precipitate the product, dissolving the precipitate in distilled water, and freeze-drying under vacuum to obtain white product, i.e. polypeptidyl diamine, and storing at a temperature below 0 ℃ in a sealed manner.
2. Preparation of p-dinitrobenzene active ester
A solution of triethylamine (0.0804mol) and p-nitrophenol (0.0804mol) in 100ml of acetone was kept at-78 ℃ at room temperature with dry ice and acetone. Sebacoyl chloride (0.04mol) in 80ml of acetone is then added dropwise to the cooled solution, stirred at-78 ℃ for 2 hours and then at room temperature overnight. Thereafter, the mixture was poured into 1000ml of distilled water to precipitate the product, which was filtered, washed thoroughly with distilled water, dried under vacuum at 50 ℃ and finally purified by recrystallization from ethyl acetate for 3 times to give needle-like off-white solid p-dinitrobenzene active ester, which was stored sealed and dried.
3. Preparation of enzyme degradable polypeptide-based polyester ammonia
The polypeptidyl diamine (1.0mmol) and the p-dinitrobenzene active ester (1mmol) were dissolved in 1.5ml of anhydrous DMAC and the solution was heated to 60 ℃ with stirring until the monomers were completely dissolved. Will ET3N (2.2mmol) was added dropwise to the solution, and the reaction was held at 80 ℃ for 16 hours to effect polymerization. Precipitating the obtained solution with cold ethyl acetate, filtering, extracting with ethyl acetate in a Soxhlet extractor for 48 hours, finally drying at 50 ℃ in vacuum to obtain white solid polypeptide-based polyesteramide, sealing, drying and storing.
4. Degradation of enzymatically degradable polypeptidyl polyesteramines
Dissolving the polypeptidyl polyesteramide in DMF to form a polymer solution with the mass fraction of 30%, removing the solvent to form a film, placing the obtained film in a PBS solution with the trypsin concentration of 0.15mg/ml, wherein the PBS solution contains 0.2mg/ml of fibroblast growth factor (VEGF), and degrading the polymer in a water bath shaker at 37 ℃ at a certain shaking speed to obtain drug-loaded nanoparticles, freezing at-19 ℃ and freeze-drying and storing at-50 ℃.
The polypeptide-based polyester ammonia-type nanoparticles prepared by the embodiment are spherical, have no obvious agglomeration phenomenon and have a stable structure; the particle size distribution is uniform, and the particle size is kept about 200 nm.
The drug loading performance of the growth factor preloaded polypeptide-based polyester ammonia type nanoparticles prepared by the embodiment can be characterized by the drug loading rate, wherein the drug loading rate is loaded with drugs/(loaded drugs + carrier mass), and the drug loading rate can be 55% by detection in combination with infrared, simple, nuclear magnetic and ultraviolet molecular methods,placing the nanoparticle and human fibroblast with pre-drug loading in a container containing 5% CO2And culturing in a constant-temperature incubator at 37 ℃ for a certain time, observing the shape and the number of the cells under an electron microscope, and thus, the number of the cells is obviously increased, so that the effect of promoting cell proliferation is verified.
Claims (7)
1. A polypeptide-based polyester ammonia-type nanoparticle characterized by: the polypeptide-based polyester ammonia is prepared by taking enzyme-degradable polypeptide-based polyester ammonia as a raw material and degrading and assembling the raw material under the action of enzyme;
wherein the chemical structural formula of the enzyme-degradable multi-peptidyl polyester ammonia is as follows:
2. A nanoparticle according to claim 1, wherein: the enzyme is an enzyme for degrading amido bond or ester bond, and is selected from trypsin, alpha-chymotrypsin, lipase or phospholipase.
3. A method for preparing the polypeptidyl polyester ammonia-type nanoparticles of claim 1, comprising:
dissolving enzyme-degradable polypeptidyl polyesteramide in an organic solvent to obtain a polyesteramide solution with the mass fraction of 5-35%, removing the solvent to form a membrane, and degrading in an enzyme-containing PBS solution water bath shaking table with the concentration of 0.05-2.0 mg/mL to obtain the polypeptidyl polyesteramide nano particles.
4. The production method according to claim 3, characterized in that: the organic solvent is DMF, DMAC, THF or CHCl3。
5. The production method according to claim 3, characterized in that: the temperature of the water bath was 37 ℃.
6. The production method according to claim 3, characterized in that: the polypeptidyl polyester ammonia-type nano particles are frozen at the temperature of 19 ℃ below zero and then freeze-dried and stored at the temperature of 50 ℃ below zero.
7. Use of the polypeptidyl polyester ammonia-type nanoparticles of claim 1 for drug loading.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811573662.2A CN109734901B (en) | 2018-12-21 | 2018-12-21 | Polypeptide-based polyester ammonia-type nano particle and preparation and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811573662.2A CN109734901B (en) | 2018-12-21 | 2018-12-21 | Polypeptide-based polyester ammonia-type nano particle and preparation and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109734901A CN109734901A (en) | 2019-05-10 |
| CN109734901B true CN109734901B (en) | 2021-05-04 |
Family
ID=66359508
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811573662.2A Active CN109734901B (en) | 2018-12-21 | 2018-12-21 | Polypeptide-based polyester ammonia-type nano particle and preparation and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109734901B (en) |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007133616A3 (en) * | 2006-05-09 | 2008-01-24 | Medivas Llc | Biodegradable water soluble polymers |
| CN101969963A (en) * | 2008-01-28 | 2011-02-09 | 那野伽利阿株式会社 | Pharmaceutical composition and combined agent |
| CN102146200A (en) * | 2011-04-19 | 2011-08-10 | 复旦大学 | Temperature-sensitive hydrogel based on chemical cross-linking gel granules and preparation method of temperature-sensitive hydrogel |
| CN102397258A (en) * | 2011-11-18 | 2012-04-04 | 中国科学院长春应用化学研究所 | Insulin carrying microsphere and preparation method thereof |
| CN102532533A (en) * | 2010-12-16 | 2012-07-04 | 国家纳米科学中心 | Poly(aspartic acid-co-lactic acid)-phosphatidyl ethanolamine graft polymer and preparation method and application thereof |
| CN102791255A (en) * | 2010-03-12 | 2012-11-21 | 帝斯曼知识产权资产管理有限公司 | Micelle compositions and process for the preparation thereof |
| CN107847398A (en) * | 2016-05-05 | 2018-03-27 | 南京三迭纪医药科技有限公司 | The pharmaceutical dosage form of control release |
| CN108743972A (en) * | 2018-06-21 | 2018-11-06 | 东华大学 | A kind of amino acidic group polyesteramine micella, preparation method and applications |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060188469A1 (en) * | 2003-10-14 | 2006-08-24 | Medivas, Llc | Vaccine delivery compositions and methods of use |
| US20070134332A1 (en) * | 2005-11-21 | 2007-06-14 | Medivas, Llc | Polymer particles for delivery of macromolecules and methods of use |
-
2018
- 2018-12-21 CN CN201811573662.2A patent/CN109734901B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007133616A3 (en) * | 2006-05-09 | 2008-01-24 | Medivas Llc | Biodegradable water soluble polymers |
| CN101969963A (en) * | 2008-01-28 | 2011-02-09 | 那野伽利阿株式会社 | Pharmaceutical composition and combined agent |
| CN102791255A (en) * | 2010-03-12 | 2012-11-21 | 帝斯曼知识产权资产管理有限公司 | Micelle compositions and process for the preparation thereof |
| CN102532533A (en) * | 2010-12-16 | 2012-07-04 | 国家纳米科学中心 | Poly(aspartic acid-co-lactic acid)-phosphatidyl ethanolamine graft polymer and preparation method and application thereof |
| CN102146200A (en) * | 2011-04-19 | 2011-08-10 | 复旦大学 | Temperature-sensitive hydrogel based on chemical cross-linking gel granules and preparation method of temperature-sensitive hydrogel |
| CN102397258A (en) * | 2011-11-18 | 2012-04-04 | 中国科学院长春应用化学研究所 | Insulin carrying microsphere and preparation method thereof |
| CN107847398A (en) * | 2016-05-05 | 2018-03-27 | 南京三迭纪医药科技有限公司 | The pharmaceutical dosage form of control release |
| CN108743972A (en) * | 2018-06-21 | 2018-11-06 | 东华大学 | A kind of amino acidic group polyesteramine micella, preparation method and applications |
Non-Patent Citations (4)
| Title |
|---|
| Biomimetic Shells Endow Sub-50 nm Nanoparticles with Ultrahigh;Xing Chen等;《ACS Appl. Mater. Interfaces》;20181010;第10卷(第40期);全文 * |
| From macro to micro to nano: the development of a novel lysine based hydrogel platform and enzyme triggered self-assembly of macro hydrogel into nanogel;De-Qun Wu等;《Journal of Materials Chemistry B》;20150109;第3卷(第11期);摘要，第2287页第2.4节，第2288页第2.9、2.12节 * |
| 一种基于赖氨酸聚酯氨新型载药纳米水凝胶;崔海春等;《东华大学学报 自然科学版》;20161231;第42卷(第6期);第853页第1.3，1.4节 * |
| 聚酯类超支化聚合物的研究现状及应用展望;王学川等;《皮革科学与工程》;20081031;第18卷(第5期);全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109734901A (en) | 2019-05-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Yadav et al. | Short to ultrashort peptide-based hydrogels as a platform for biomedical applications | |
| Fonseca et al. | Biodegradable poly (ester amide) s–A remarkable opportunity for the biomedical area: Review on the synthesis, characterization and applications | |
| Mallakpour et al. | Progress in synthetic polymers based on natural amino acids | |
| US4892733A (en) | Biodegradable synthesis polypeptide and its therapeutic use | |
| Ma et al. | Development of ionic strength/pH/enzyme triple-responsive zwitterionic hydrogel of the mixed l-glutamic acid and l-lysine polypeptide for site-specific drug delivery | |
| Tsuchiya et al. | Chemoenzymatic synthesis of polypeptides for use as functional and structural materials | |
| Liu et al. | Preparation and applications of peptide-based injectable hydrogels | |
| Gupta et al. | Amino acid derived biopolymers: Recent advances and biomedical applications | |
| Arakawa et al. | Polymer design and development | |
| Binaymotlagh et al. | Peptide-based hydrogels: New materials for biosensing and biomedical applications | |
| Yan et al. | Fabrication of injectable hydrogels based on poly (l-glutamic acid) and chitosan | |
| AU2005257078B2 (en) | Hydrogels of hyaluronic acid and alpha, beta-polyaspartylhydrazide and their biomedical and pharmaceutical uses | |
| Guo et al. | Biodegradation of unsaturated poly (ester-amide) s and their hydrogels | |
| CN109734900B (en) | Enzyme-degradable polypeptide-based polyester ammonia and preparation method and application thereof | |
| Wu et al. | Redox-responsive supramolecular hydrogel based on 10-hydroxy camptothecin-peptide covalent conjugates with high loading capacity for drug delivery | |
| Wu et al. | Structural design and antimicrobial properties of polypeptides and saccharide–polypeptide conjugates | |
| CN107459654B (en) | Simulated antibacterial peptide copolymer and preparation method thereof | |
| Arias et al. | Tailored recombinant elastin-like polymers for advanced biomedical and nano (bio) technological applications | |
| Zavradashvili et al. | Artificial polymers made of α-amino acids-poly (amino acid) s, pseudo-poly (amino acid) s, poly (depsipeptide) s, and pseudo-proteins | |
| Fan et al. | Poly (glutamic acid) hydrogels crosslinked via native chemical ligation | |
| Jalilpour et al. | A simple route for preparation of pH-sensitive hydrogels by using egg white proteins in alginate scaffold for the encapsulation of probiotics | |
| CN114133435A (en) | Elastin-like polypeptide and application | |
| CN109517162B (en) | Injectable hydrogel and preparation method thereof | |
| CN109734901B (en) | Polypeptide-based polyester ammonia-type nano particle and preparation and application thereof | |
| Watanabe et al. | Chemoenzymatic polymerization of l-serine ethyl Ester in aqueous media without side-group protection |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |