CN114957706B - Vesicle with stability and permeability as well as preparation method and application thereof - Google Patents
Vesicle with stability and permeability as well as preparation method and application thereof Download PDFInfo
- Publication number
- CN114957706B CN114957706B CN202210572208.5A CN202210572208A CN114957706B CN 114957706 B CN114957706 B CN 114957706B CN 202210572208 A CN202210572208 A CN 202210572208A CN 114957706 B CN114957706 B CN 114957706B
- Authority
- CN
- China
- Prior art keywords
- vesicle
- permeability
- vesicles
- preparation
- monomer
- 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
- 230000035699 permeability Effects 0.000 title claims abstract description 30
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 239000000178 monomer Substances 0.000 claims abstract description 32
- 238000001338 self-assembly Methods 0.000 claims abstract description 11
- 230000002209 hydrophobic effect Effects 0.000 claims abstract description 7
- 150000002433 hydrophilic molecules Chemical class 0.000 claims abstract description 6
- 238000011065 in-situ storage Methods 0.000 claims abstract description 5
- OXBLVCZKDOZZOJ-UHFFFAOYSA-N 2,3-Dihydrothiophene Chemical compound C1CC=CS1 OXBLVCZKDOZZOJ-UHFFFAOYSA-N 0.000 claims abstract description 3
- 238000012650 click reaction Methods 0.000 claims abstract 2
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 16
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 15
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 13
- 239000012074 organic phase Substances 0.000 claims description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- BHELZAPQIKSEDF-UHFFFAOYSA-N allyl bromide Chemical compound BrCC=C BHELZAPQIKSEDF-UHFFFAOYSA-N 0.000 claims description 6
- WXZMFSXDPGVJKK-UHFFFAOYSA-N pentaerythritol Chemical compound OCC(CO)(CO)CO WXZMFSXDPGVJKK-UHFFFAOYSA-N 0.000 claims description 6
- JRMUNVKIHCOMHV-UHFFFAOYSA-M tetrabutylammonium bromide Chemical compound [Br-].CCCC[N+](CCCC)(CCCC)CCCC JRMUNVKIHCOMHV-UHFFFAOYSA-M 0.000 claims description 6
- 102000004190 Enzymes Human genes 0.000 claims description 5
- 108090000790 Enzymes Proteins 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 239000007864 aqueous solution Substances 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical group SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 claims description 3
- 239000011259 mixed solution Substances 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- 238000001291 vacuum drying Methods 0.000 claims description 3
- 239000003054 catalyst Substances 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 238000010438 heat treatment Methods 0.000 claims description 2
- 238000003756 stirring Methods 0.000 claims description 2
- 238000005406 washing Methods 0.000 claims description 2
- 238000007664 blowing Methods 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 4
- 239000002356 single layer Substances 0.000 abstract description 4
- 230000006698 induction Effects 0.000 abstract description 2
- 230000001699 photocatalysis Effects 0.000 abstract 1
- 238000007146 photocatalysis Methods 0.000 abstract 1
- 102000016938 Catalase Human genes 0.000 description 22
- 108010053835 Catalase Proteins 0.000 description 19
- 230000015572 biosynthetic process Effects 0.000 description 13
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 11
- 239000003642 reactive oxygen metabolite Substances 0.000 description 8
- 230000008569 process Effects 0.000 description 7
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 6
- 230000003197 catalytic effect Effects 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- 239000002086 nanomaterial Substances 0.000 description 6
- 239000002245 particle Substances 0.000 description 6
- 238000003786 synthesis reaction Methods 0.000 description 6
- 230000002427 irreversible effect Effects 0.000 description 5
- 239000012528 membrane Substances 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 238000003556 assay Methods 0.000 description 4
- 238000004132 cross linking Methods 0.000 description 4
- 238000005538 encapsulation Methods 0.000 description 4
- 230000001965 increasing effect Effects 0.000 description 4
- 229920000642 polymer Polymers 0.000 description 4
- 238000006116 polymerization reaction Methods 0.000 description 4
- 229920000575 polymersome Polymers 0.000 description 4
- 239000011541 reaction mixture Substances 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- 150000003573 thiols Chemical group 0.000 description 4
- UOWNEWCMPHICQH-UHFFFAOYSA-N 1,2-bis(2-bromoethoxy)ethane Chemical compound BrCCOCCOCCBr UOWNEWCMPHICQH-UHFFFAOYSA-N 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- 238000003917 TEM image Methods 0.000 description 3
- 238000002296 dynamic light scattering Methods 0.000 description 3
- 230000003834 intracellular effect Effects 0.000 description 3
- 239000003208 petroleum Substances 0.000 description 3
- 238000000235 small-angle X-ray scattering Methods 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- KIUMMUBSPKGMOY-UHFFFAOYSA-N 3,3'-Dithiobis(6-nitrobenzoic acid) Chemical compound C1=C([N+]([O-])=O)C(C(=O)O)=CC(SSC=2C=C(C(=CC=2)[N+]([O-])=O)C(O)=O)=C1 KIUMMUBSPKGMOY-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241000282326 Felis catus Species 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- 102000004142 Trypsin Human genes 0.000 description 2
- 108090000631 Trypsin Proteins 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000004630 atomic force microscopy Methods 0.000 description 2
- 239000012267 brine Substances 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 238000000354 decomposition reaction Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 150000004662 dithiols Chemical class 0.000 description 2
- 239000003995 emulsifying agent Substances 0.000 description 2
- 238000003818 flash chromatography Methods 0.000 description 2
- 239000000543 intermediate Substances 0.000 description 2
- 230000008520 organization Effects 0.000 description 2
- 238000010926 purge Methods 0.000 description 2
- 238000010992 reflux Methods 0.000 description 2
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 2
- 230000002194 synthesizing effect Effects 0.000 description 2
- 239000006188 syrup Substances 0.000 description 2
- 235000020357 syrup Nutrition 0.000 description 2
- UMGDCJDMYOKAJW-UHFFFAOYSA-N thiourea Chemical compound NC(N)=S UMGDCJDMYOKAJW-UHFFFAOYSA-N 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 238000004627 transmission electron microscopy Methods 0.000 description 2
- 239000012588 trypsin Substances 0.000 description 2
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 description 2
- 230000007332 vesicle formation Effects 0.000 description 2
- VQVUBYASAICPFU-UHFFFAOYSA-N (6'-acetyloxy-2',7'-dichloro-3-oxospiro[2-benzofuran-1,9'-xanthene]-3'-yl) acetate Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(Cl)=C(OC(C)=O)C=C1OC1=C2C=C(Cl)C(OC(=O)C)=C1 VQVUBYASAICPFU-UHFFFAOYSA-N 0.000 description 1
- -1 1-mercapto-3, 6-dioxepane Chemical compound 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 125000000022 2-aminoethyl group Chemical group [H]C([*])([H])C([H])([H])N([H])[H] 0.000 description 1
- 239000000592 Artificial Cell Substances 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 241000283891 Kobus Species 0.000 description 1
- PHSPJQZRQAJPPF-UHFFFAOYSA-N N-alpha-Methylhistamine Chemical compound CNCCC1=CN=CN1 PHSPJQZRQAJPPF-UHFFFAOYSA-N 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Natural products NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000003929 acidic solution Substances 0.000 description 1
- 125000005336 allyloxy group Chemical group 0.000 description 1
- 229920000469 amphiphilic block copolymer Polymers 0.000 description 1
- 238000000149 argon plasma sintering Methods 0.000 description 1
- 238000000089 atomic force micrograph Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003012 bilayer membrane Substances 0.000 description 1
- 229920001400 block copolymer Polymers 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000010382 chemical cross-linking Methods 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000008602 contraction Effects 0.000 description 1
- 230000008094 contradictory effect Effects 0.000 description 1
- 239000003431 cross linking reagent Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000002088 nanocapsule Substances 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- 238000011056 performance test Methods 0.000 description 1
- 230000002688 persistence Effects 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000012429 reaction media Substances 0.000 description 1
- 230000036632 reaction speed Effects 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 238000001464 small-angle X-ray scattering data Methods 0.000 description 1
- 238000001370 static light scattering Methods 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- ZIBGPFATKBEMQZ-UHFFFAOYSA-N triethylene glycol Chemical compound OCCOCCOCCO ZIBGPFATKBEMQZ-UHFFFAOYSA-N 0.000 description 1
- 238000002371 ultraviolet--visible spectrum Methods 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
- A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers
- A61K9/1273—Polymersomes; Liposomes with polymerisable or polymerised bilayer-forming substances
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G75/00—Macromolecular compounds obtained by reactions forming a linkage containing sulfur with or without nitrogen, oxygen, or carbon in the main chain of the macromolecule
- C08G75/02—Polythioethers
- C08G75/04—Polythioethers from mercapto compounds or metallic derivatives thereof
- C08G75/045—Polythioethers from mercapto compounds or metallic derivatives thereof from mercapto compounds and unsaturated compounds
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
- C12N11/089—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0065—Oxidoreductases (1.) acting on hydrogen peroxide as acceptor (1.11)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y111/00—Oxidoreductases acting on a peroxide as acceptor (1.11)
- C12Y111/01—Peroxidases (1.11.1)
- C12Y111/01006—Catalase (1.11.1.6)
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2381/00—Characterised by the use of macromolecular compounds obtained by reactions forming in the main chain of the macromolecule a linkage containing sulfur with or without nitrogen, oxygen, or carbon only; Polysulfones; Derivatives of such polymers
- C08J2381/02—Polythioethers; Polythioether-ethers
Abstract
The invention discloses a vesicle with stability and permeability, and a preparation method and application thereof. The preparation method comprises the following steps: under the condition of no need of monomer self-assembly in advance or template induction, under the photocatalysis effect, the hydrophobic monomer and the hydrophilic monomer are gradually polymerized to form double-headed hydrophilic molecules through thiol-ene click reaction in an environment with proper polarity, and then the vesicle with both stability and permeability can be prepared by in-situ self-assembly. According to the invention, the vesicle is constructed by in-situ generation and covalent bond self-assembly of the double-headed parent molecule, and the prepared vesicle is an ultrathin single-layer vesicle and has excellent stability and permeability.
Description
Technical Field
The invention belongs to the technical field of vesicle preparation, and particularly relates to a vesicle with stability and permeability, and a preparation method and application thereof.
Background
Vesicles are self-assembled from phospholipids of natural origin, providing an impermeable compartment for eukaryotic cell life activities. Artificially synthesized vesicle systems have attracted considerable interest in biomedical, nanoreactors, artificial organelles, and the like over the last several decades. In addition to natural lipids, synthetic amphiphilic block copolymers are the most common materials for constructing vesicle systems. However, polymer vesicles are very limited in practical use due to their poor stability.
The most straightforward method to improve vesicle stability is covalent cross-linking of dynamic self-assembled nanostructures. However, this method usually requires multiple steps to complete during the synthesis process, and changes the morphology of the nanostructures, which also has a certain impact on the precipitation of aggregates. To date, direct synthesis of vesicles using irreversible covalent bonds without the aid of any pre-organization, templates or emulsifiers remains a difficult challenge. In recent years Kim and co-workers have found that irreversible covalent bonds can be used to synthesize a variety of nanostructured materials with well-defined structures. This is in contrast to self-assembly by thermodynamic control, where the polymeric nanocapsules need to be covalently crosslinked by pre-organized rigid building blocks during the preparation process to form a dimensionally stable covalent spherical nanostructure. However, as is well known, no method for directly synthesizing covalent vesicles having high stability and high permeability by using flexible monomers with high degrees of freedom to react with each other has been developed. . The reason is that functional groups with high degrees of freedom and polyvalent nature may cause an unoriented cross-linked structure during the flexible monomer interaction and also that no controlled sequential polymerization and shape persistence induced metastable morphology transformation occurs.
In addition, stability and permeability are two important properties in practical applications of vesicles. To solve the problem of vesicle permeability, researchers all around the world have performed a lot of work to find a method for improving the permeability through physical or chemical modification. However, the stability and permeability of vesicles are often contradictory properties, i.e. it is difficult to increase the permeability again while increasing the stability, and the permeability becomes even worse. Also, increasing vesicle permeability may decrease vesicle stability. To solve this problem, researchers have generally used chemical crosslinking to increase the stability of vesicles and then to increase their permeability by modulating the bilayer membrane of the vesicles. Among them, covalent crosslinking is the most straightforward method of improving vesicle stability, but as previously mentioned, crosslinking can improve stability, but often results in reduced permeability. Furthermore, the direct synthesis of vesicles using irreversible covalent bonds remains a difficult challenge to date without the aid of any pre-organization, templates or emulsifiers. Thus, there is an urgent need for a rapid and simple process to prepare vesicles that have both excellent stability and permeability.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a vesicle with stability and permeability, and a preparation method and application thereof, which can effectively solve the problem that the stability and the permeability of the vesicle prepared by the prior art cannot be taken into consideration.
In order to achieve the above purpose, the technical scheme adopted by the invention for solving the technical problems is as follows:
the vesicle with both stability and permeability is prepared through the polymerization of hydrophobic monomer and hydrophilic monomer in proper polarity environment via thiol-ene clicking reaction to form double-headed hydrophilic molecule and the in-situ self-assembly of the double-headed hydrophilic molecule under the condition of no requirement of self-assembly of monomer or template induction.
Further, the vesicle formation process is:
adding a hydrophobic monomer and a hydrophilic monomer into a mixed solution of methanol and water with the polarity of 7-7.5, purging with nitrogen, irradiating for 60-80 min at room temperature, dialyzing for 2-3 days, concentrating under reduced pressure, and vacuum drying to obtain the vesicle.
Further, the polarity of the mixed solution is 7.2 to 7.32.
Further, the hydrophobic monomer is tetra-o-allylpentaerythritol, and the chemical structural formula is as follows:
further, the preparation method of the tetra-o-allylpentaerythritol comprises the following steps:
dissolving pentaerythritol in a 33% NaOH aqueous solution, adding tetrabutylammonium bromide, heating to 40-50 ℃, adding allyl bromide in 1h, stirring for 12-15 h at room temperature, adding toluene, extracting, washing an organic phase, and concentrating and purifying the organic phase.
Further, the molar ratio of pentaerythritol, tetrabutylammonium bromide and allyl bromide is 20-22:4-6.2:100-118.
Further, the preparation method of the tetra-o-allylpentaerythritol comprises the following steps:
pentaerythritol (6.3 g,22 mmol) was dissolved in 33% aqueous NaOH (50 mL). Tetrabutylammonium bromide (2 g,6.2 mmol) was added, the temperature was raised to 40 ℃, and then allyl bromide (10 ml,118 mmol) was added dropwise over 1 h. The reaction mixture was vigorously stirred at room temperature for 12h until stirred well. Toluene (100 mL) was added, the organic phase was washed with water, concentrated in vacuo, and purified by flash chromatography (toluene: ethyl acetate, 20:1) to give the allylated product (3.84 g,13mmol, 59%) as a colorless syrup
Further, the hydrophilic monomer is a thiol having hydrophilicity; the thiol contains at least 2 mercapto groups.
Further, the hydrophilic monomer is dithiothreitol or 1-mercapto-3, 6-dioxepane.
After uv irradiation, particles with an average particle size of 5nm were detected starting from 2.5 minutes and the internal particle size increased rapidly to about 100nm in ten minutes, then increased slowly to the final particle size (-125 nm), after which it remained unchanged. The particle size growth process is fully in line with the concept of double-headed affinity molecule formation and self-assembly. The initial phase of the reaction (2.5 min) can be assumed to be the formation process of the bipartite parent molecule, and as soon as the concentration of the bipartite parent molecule reaches the requirement of self-assembly, the pre-organized covalent vesicle intermediates will start to grow. Likewise, TEM also monitored the process of this vesicle formation (fig. 2).
No pre-tissue structure was observed before uv irradiation, after which small plaques (2-5 nm) appeared at 2.5 minutes (fig. 2 a), which then began to curl, forming bowl-like nanostructures, and further transformation and growth in the transverse direction was completed within a few minutes (fig. 2 b-d). A reasonable integration process and a large number of covalent vesicles were observed in the following 10-15 minutes (fig. 2 d-f).
Vesicles prepared by the method.
The application of the vesicle in preparing a drug carrier, a catalyst, a chemical sensor, a nano-reactor and enzyme immobilization.
The invention has the beneficial effects that:
according to the invention, the vesicle is constructed by using the double-head amphiphilic molecule and the flexible monomer through the formation of irreversible covalent bonds, and the prepared vesicle is an ultrathin single-layer vesicle and has excellent stability and permeability.
Drawings
FIG. 1 shows the performance test of vesicles prepared according to the invention; wherein a is a TEM image; b is an AFM image; c is SAXS pattern; d is the vesicle size and PDI;
FIG. 2 is an animated image and TEM image of the vesicles prepared according to the invention at various time points during polymerization;
FIG. 3 shows the results of an enzyme encapsulation assay; a is an experimental device established by connecting a pump with a plastic model system; b is the effect of shear forces on the staged release of the contents encapsulated in vesicles and in commercially available polymers; c is free and coated CAT catalytic H under 40Pa shear stress and trypsin condition 2 O 2 Is a comparison of degradation kinetics of (2); d is the superposition of fluorescent images (first row), bright-field images (second row) and fluorescent images combined with bright-field (third row) by different post-treatment intracellular ROS (reactive oxygen species) levels; e is ROS level detection.
Detailed Description
The following description of the embodiments of the present invention is provided to facilitate understanding of the present invention by those skilled in the art, but it should be understood that the present invention is not limited to the scope of the embodiments, and all the inventions which make use of the inventive concept are protected by the spirit and scope of the present invention as defined and defined in the appended claims to those skilled in the art.
Example 1
1. Synthesis of Tetrao-allyl pentaerythritol (monomer 1)
Pentaerythritol (6.3 g,22 mmol) was dissolved in 33% aqueous NaOH (50 mL). Tetrabutylammonium bromide (2 g,6.2 mmol) was added, the temperature was raised to 40 ℃, and then allyl bromide (10 ml,118 mmol) was added dropwise over 1 h. The reaction mixture was vigorously stirred at room temperature for 12h until stirred well. Toluene (100 mL) was added, the organic phase was washed with water, concentrated in vacuo, and purified by flash chromatography (toluene: ethyl acetate, 20:1) to give the allylated product (3.84 g,13mmol, 59%) as a colorless syrup. The nuclear magnetic data are as follows: 1 H NMR(400MHz,CDCl 3 ) Delta 3.91 (dd, j=5.9 hz, 2H), 3.62 (dd, j=7.7 hz, 2H), 3.08-3.12 (m, 2H), 2.88-2.91 (m, 2H), 2.05 (s, 1H). The reaction formula is as follows:
2. dithiothreitol (monomer 2) is a commercially available product.
3. Construction of monomer 3
Synthesis of 1, 2-bis (2-bromoethoxy) ethane: to a solution of triethylene glycol (1.5 g,10 mmol) in 20mL dry dichloromethane was added dropwise phosphorus tribromide (8.1 g,30 mmol) and the reaction mixture was stirred at reflux for 24h. The reaction solution was then diluted with dichloromethane (40 mL) and neutralized with saturated aqueous NaHCO 3. The organic phase was collected, washed with brine and dried over anhydrous magnesium sulfate. The solvent was removed in vacuo and the residue was purified by column chromatography (petroleum ether: ethyl acetate=10:1) to give 1, 2-bis (2-bromoethoxy) ethane (1.9 g, 69%) as a yellow oil. 1H NMR (400 MHz, CDCl 3) δ:3.44 (t, J=1.2 Hz, 4H), 3.65 (s, 4H), 3.79 (t, J=1.2 Hz, 4H).
Synthesis of 1, 8-dimercapto-3, 6-dioxepane: 1, 2-bis (2-bromoethoxy) ethane (304 mg,1.0 mmol) and thiourea (365.8 mg,4.8 mmol) were dissolved in 95% ethanol (30 mL). The reaction mixture was stirred under reflux for 3h, then 20mL of sodium hydroxide solution (1.0 mol/L) was added andreflux was continued for 2h. The aqueous layer was separated and acidified (ph=1) by addition of dilute hydrochloric acid (2.0 mol/L). The acidic solution was then extracted with petroleum ether (3X 30 mL). The organics were combined, washed with brine and dried over anhydrous magnesium sulfate. The solvent was removed in vacuo and the residue was purified by column chromatography (petroleum ether: ethyl acetate=3:1) to give the compound as a colourless oil (156 mg, 85.7%). The nuclear magnetic data are as follows: 1 H NMR(400MHz,CDCl 3 ) Delta 3.91 (dd, j=5.9 hz, 2H), 3.62 (dd, j=7.7 hz, 2H), 3.08-3.12 (m, 2H), 2.88-2.91 (m, 2H), 2.05 (s, 1H). The reaction formula is as follows:
example 2
1. Preparation of vesicles from monomer 1 and monomer 2
Tetrao-allylpentaerythritol (monomer 1) and dithiol (monomer 2) were added to MeOH/H 2 O(v/v＝5:1，P AB =7.25) in solution, mixed in a ratio of 1:2 (allyloxy: thiol=1:1), and after purging with nitrogen and irradiation with 365nm uv light at room temperature for 60 minutes, then dialysis in water for two days, vacuum drying after concentrating under reduced pressure, the vesicle was obtained (yield: 98%).
The morphology and size of the covalent vesicles can be characterized by spectroscopic, light scattering and imaging techniques (fig. 1), and from fig. 1 it can be seen that the size and PDI are independent of concentration, which confirms the cross-linking nature of the vesicles, indicating that the vesicles have good stability (d in fig. 1).
When studied using Transmission Electron Microscopy (TEM) and Atomic Force Microscopy (AFM), synthetic vesicles with nanoscale colloidal particle properties were first presented (fig. 1a, b) and the diameters of the vesicles were very consistent with the average hydrodynamic diameter (125±4 nm) observed with Dynamic Light Scattering (DLS) (table 1).
A combination of dynamic and static light scattering studies can also measure the radius of gyration (rg=66 nm), the hydrodynamic radius (rh=63 nm) and the ratio of Rg/Rh (-1.05) of the vesicles, which can reflect the hollow sphere properties of the vesicles.
In addition, TEM micrographs showed a hollow interior surrounded by an ultrathin membrane (fig. 1 a), again confirming that the vesicles have hollow sphere properties by small angle X-ray scattering (SAXS) analysis of the vesicles (fig. 1 c).
The average thickness of vesicles (2.8.+ -. 0.4 nm) calculated by SAXS analysis was consistent with the size of the double-headed affinity intermediate, indicating the formation of a monolayer. From this, it is clear that synthetic vesicles are nanostructures composed of a monolayer film with stable hollow sphere properties. And synthetic vesicles become more stable due to the formation of multiple C-S covalent bonds, which is a distinct difference from self-assembled artificial vesicle systems (fig. 1 d).
2. When monomer 1 is crosslinked with thiol monomers that do not contain two hydrophilic hydroxyl groups, no well-defined covalent vesicles can be obtained due to the lack of formation and self-assembly of double-headed hydrophilic molecules. If the dithiol cross-linking agent is converted into monomer 3 with an ethylene glycol segment in between, then well-defined covalent vesicles can be regenerated within an optimized polarity range.
Example 3
1. Vesicle permeability assay
Stability and permeability are two important properties of vesicles in practical applications, with membrane permeability being one of the most important properties that determine their potential application. Determination of organophosphorus reagent loaded PP using a reaction Rate assay for ADNB substrates (also referred to as Ellman reagent) 3 Membrane permeability of vesicles of TS.
Under the experimental conditions of (k)>10M -1 ·s -1 ) In the case of DTNB and free PP 3 The reaction speed of TS is very fast. Adding ADNB to load PP 3 In aqueous solution of vesicles of TS. The substrate must penetrate into the vesicles and the PP 3 TS reaction and conversion to colored products. These colored products are then detected by the ultraviolet-visible spectrum. At room temperature, 0.5mL of PP containing vesicles was used 3 TS solution was mixed with 2.5mL of ADNB aqueous solution (4.5. Mu.M) and the reaction rate was measured in a 1cm silica glass reaction cup. Absorbance at 412nm was recorded every 20s and the permeability of the vesicles was determined (see table 1).
Table 1 comparison of the permeabilities of vesicles
As shown in Table 1, by monitoring the highly developed product 5-thio-2-nitrobenzoate (TNB) (ε=14150M at 412 nm) -1 ·cm -1 ) To measure the permeability of the synthetic vesicles, the substrate was 5,5- (2-aminoethyl) -dithio-2-nitrobenzoate (ADNB). All synthetic vesicles have very high permeability to small organic molecules (up to 57.9.+ -. 1.8 nm.s compared to block copolymer assembled polymer vesicles -1 ). And reaction medium (P) AB 、MeOH/H 2 The higher the polarity of O), the larger the hydrodynamic diameter of the synthetic vesicle and the greater the osmotic capacity.
It can be seen that during the irreversible polymerization, the polarity (P AB (polarity of solvent), meOH/H 2 O ratio) plays an important role in synthesizing vesicles and controlling the size of vesicles, and only at P AB In the range of 7.20-7.32, distinct vesicles can be formed.
Example 4
The enzyme encapsulation performance is detected as follows:
by exploring the encapsulation of Catalase (CAT) using synthetic vesicles, CAT can be encapsulated by in situ bipitch parent molecule formation and self-assembly by simply mixing with monomer 1 and monomer 2. .
With the CAT-loaded vesicles, we further used a detection instrument to detect the stability of CAT vascular simulations at different pressures (fig. 3 a). To highlight the advantages of vesicles, CAT@1V (CAT@vesicles) and CAT@polymersome (CAT@polymer vesicles) systems were measured for CAT release under different shear stresses (0-40 Pa). The CAT release rate of the cat@polymersome system, which is a standard on the market, is very high, whereas cat@1v shows that the CAT release rate of the crosslinked membrane tight vesicles is lower than 5% even under simulated shear stress (40 Pa) of arterial contraction. (FIG. 3 b)
The CAT@1V system achieved good stability after encapsulation, and the CAT vs H was subsequently evaluated by FOX assay 2 O 2 Catalytic activity of decomposition. We obtained higher Kobs than CAT@1V system, CAT@polymersome system and CAT alone (0.0473 s -1 ) This suggests that vesicles not only efficiently transport substrates and products across the membrane, but also enhance the catalytic activity of CAT. Furthermore, the protection of trypsin by encapsulated CAT is better than other systems (fig. 3 c). .
In organisms, the production of Reactive Oxygen Species (ROS) above critical levels may cause oxidative damage to intracellular biomolecules, thereby inducing a number of diseases. To detect hydrogen peroxide (H) in CAT to life systems 2 O 2 ) Catalytic Activity of decomposition Using 2',7' -dichlorofluorescein diacetate (DCFH-DA) as fluorescent Probe, H was introduced 2 O 2 The treated HUVEC cells were subjected to intracellular ROS studies.
As shown in fig. 3d, H 2 O 2 The treated cells produced intense fluorescence due to excessive ROS. However, when cells were incubated with cat@1v, the fluorescence intensity was significantly reduced, as low as that observed in normal cells after 128min of release. This ability to return to normal ROS concentrations is due to the stable catalytic properties of cat@1v in living cells. In contrast, while cat@polymersome system and CAT also catalyze this reaction, the catalytic efficiency was low due to the low stability and activity of the enzymes in living cells (fig. 3 e).
Claims (5)
1. A preparation method of vesicles with stability and permeability is characterized in that hydrophobic monomers and hydrophilic monomers are polymerized through thiol-ene click reaction to form double-headed hydrophilic molecules, and then the double-headed hydrophilic molecules are subjected to in-situ self-assembly;
the vesicle is formed by the following steps:
adding a hydrophobic monomer and a hydrophilic monomer into a mixed solution of methanol and water with the polarity of 7.2-7.32, blowing with nitrogen, illuminating for 10-80 min at room temperature, dialyzing for 2-3 days, concentrating under reduced pressure, and vacuum drying to obtain the vesicle; the hydrophobic monomer is tetra-o-allylpentaerythritol, and the chemical structural formula is as follows:the method comprises the steps of carrying out a first treatment on the surface of the The hydrophilic monomer is dithiothreitol or 1, 8-dimercapto-3, 6-dioxepane.
2. The preparation method according to claim 1, wherein the preparation method of the tetra-o-allylpentaerythritol comprises the following steps:
dissolving pentaerythritol in a 33% NaOH aqueous solution, adding tetrabutylammonium bromide, heating to 40-50 ℃, adding allyl bromide in 1h, stirring for 12-15 h at room temperature, adding toluene, extracting, washing an organic phase, and concentrating and purifying the organic phase.
3. The method according to claim 2, wherein the molar ratio of pentaerythritol, tetrabutylammonium bromide and allyl bromide is 20-22:4-6.2:100-118.
4. A vesicle prepared by the method of any one of claims 1 to 3.
5. Use of the vesicle of claim 4 for the preparation of a drug carrier, a catalyst, a chemical sensor, a nanoreactor, and an enzyme immobilization.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210572208.5A CN114957706B (en) | 2022-05-24 | 2022-05-24 | Vesicle with stability and permeability as well as preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210572208.5A CN114957706B (en) | 2022-05-24 | 2022-05-24 | Vesicle with stability and permeability as well as preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114957706A CN114957706A (en) | 2022-08-30 |
| CN114957706B true CN114957706B (en) | 2024-03-12 |
Family
ID=82955033
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210572208.5A Active CN114957706B (en) | 2022-05-24 | 2022-05-24 | Vesicle with stability and permeability as well as preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114957706B (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6835394B1 (en) * | 1999-12-14 | 2004-12-28 | The Trustees Of The University Of Pennsylvania | Polymersomes and related encapsulating membranes |
| CN101443048A (en) * | 2004-04-20 | 2009-05-27 | 德瑞迪克纳米科技公司 | Dendritic polymers with enhanced amplification and interior functionality |
| CN102336996A (en) * | 2010-06-22 | 2012-02-01 | 四川大学 | Time-temperature color-changing indicator and application and using method thereof |
| CN102657873A (en) * | 2012-05-21 | 2012-09-12 | 苏州大学 | Vesicles consisting of amphiphilic polymer and application of vesicles |
| CN103588940A (en) * | 2013-11-07 | 2014-02-19 | 中国科学技术大学 | Amphipathic block polymer, polymersome, and preparation method and application of polymersome |
| CN107099007A (en) * | 2017-04-24 | 2017-08-29 | 浙江理工大学 | Polymer composite vesica of Amphipathilic block polymer, light and pH double-responses and its preparation method and application |
| CN109475573A (en) * | 2016-06-13 | 2019-03-15 | 珀杜研究基金会 | Responsiveness elastomeric polymer and its preparation and application |
| US10696985B1 (en) * | 2016-06-06 | 2020-06-30 | Vanderbilt University | Reversibly crosslinked endosomolytic polymer vesicles for cytosolic drug delivery |
| CN113321596A (en) * | 2021-06-08 | 2021-08-31 | 四川大学华西医院 | Photochromic controllable permeable small-molecule cross-linked vesicle and preparation method and application thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110189284A1 (en) * | 2009-12-21 | 2011-08-04 | University Of Bristol | Peptide Fibres |
| US8790621B2 (en) * | 2010-04-12 | 2014-07-29 | Iowa State University Research Foundation, Inc. | Nanoparticles and nanoparticle compositions |
| EP2911701A4 (en) * | 2012-10-23 | 2016-06-08 | Univ Johns Hopkins | Novel self-assembling drug amphiphiles and methods for synthesis and use |
-
2022
- 2022-05-24 CN CN202210572208.5A patent/CN114957706B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6835394B1 (en) * | 1999-12-14 | 2004-12-28 | The Trustees Of The University Of Pennsylvania | Polymersomes and related encapsulating membranes |
| CN101443048A (en) * | 2004-04-20 | 2009-05-27 | 德瑞迪克纳米科技公司 | Dendritic polymers with enhanced amplification and interior functionality |
| CN102336996A (en) * | 2010-06-22 | 2012-02-01 | 四川大学 | Time-temperature color-changing indicator and application and using method thereof |
| CN102657873A (en) * | 2012-05-21 | 2012-09-12 | 苏州大学 | Vesicles consisting of amphiphilic polymer and application of vesicles |
| CN103588940A (en) * | 2013-11-07 | 2014-02-19 | 中国科学技术大学 | Amphipathic block polymer, polymersome, and preparation method and application of polymersome |
| US10696985B1 (en) * | 2016-06-06 | 2020-06-30 | Vanderbilt University | Reversibly crosslinked endosomolytic polymer vesicles for cytosolic drug delivery |
| CN109475573A (en) * | 2016-06-13 | 2019-03-15 | 珀杜研究基金会 | Responsiveness elastomeric polymer and its preparation and application |
| CN107099007A (en) * | 2017-04-24 | 2017-08-29 | 浙江理工大学 | Polymer composite vesica of Amphipathilic block polymer, light and pH double-responses and its preparation method and application |
| CN113321596A (en) * | 2021-06-08 | 2021-08-31 | 四川大学华西医院 | Photochromic controllable permeable small-molecule cross-linked vesicle and preparation method and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| Azobenzene-Based Cross-Linked Small-Molecule Vesicles for Precise Oxidative Damage Treatments Featuring Controlled and Prompt Molecular Release;Yongchao Yao等;chemistry of materials;20210913;第33卷;第7357-7366页 * |
| Thiol–acrylate based vitrimers: From their structure–property relationship to the additive manufacturing of self-healable soft active devices;Usman Shaukat等;Polymer;20210822;第1-11页 * |
| 两亲性嵌段共聚物的自组装及其在生物医学上的应用进展;李汉红 等;胶体与聚合物;20190930;第37卷(第3期);第138-141页 * |
| 高分子囊泡渗透性与微结构协同调控;姚陈志;汪枭睿;胡进明;刘世勇;;高分子学报;20190426(06);第21-34页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114957706A (en) | 2022-08-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Jiang et al. | Smart pH-responsive polymer-tethered and Pd NP-loaded NMOF as the Pickering interfacial catalyst for one-pot cascade biphasic reaction | |
| Zou et al. | Clicking well‐defined biodegradable nanoparticles and nanocapsules by UV‐induced thiol‐ene cross‐linking in transparent miniemulsions | |
| Chen et al. | Designing 3D Biological Surfaces via the Breath‐Figure Method | |
| Lockhart et al. | Electron beam lithography of poly (glycidol) nanogels for immobilization of a three-enzyme cascade | |
| Tang et al. | Facile aqueous-phase synthesis of multi-responsive nanogels based on polyetheramines and bisepoxide | |
| Kazybayeva et al. | Thiol-ene “click reactions” as a promising approach to polymer materials | |
| Modigunta et al. | Pore-selective modification of the honeycomb-patterned porous polystyrene film with poly (N-isopropylacrylamide) and application for thermo-responsive smart material | |
| Cao et al. | Fabrication of pore-selective carboxyl group functionalized polyimide honeycomb-patterned porous films using KOH humidity | |
| CN105801921B (en) | A kind of cellulose esterification derivative ordered porous membrane material and preparation method thereof | |
| CN114957706B (en) | Vesicle with stability and permeability as well as preparation method and application thereof | |
| US8546488B2 (en) | Cleavable block copolymers, functionalized nanoporous thin films and related methods of preparation | |
| Qi et al. | Breaking pore size limit of metal—organic frameworks: Bio-etched ZIF-8 for lactase immobilization and delivery in vivo | |
| You et al. | Recent advances on the construction of multidimensional polydopamine-based nanostructures | |
| CN106957417A (en) | A kind of method that galapectite graphene oxide complex catalysts prepare poly epsilon caprolactone lactone | |
| WO2006080849A2 (en) | Polyisocyanide polymersomes | |
| CN106047852B (en) | A kind of flower ball-shaped Co3(PO4)2/ chloroperoxidase hybridized nanometer reactor and its application | |
| Ahmadi et al. | Synthesis and structural characterization of lignin/silica hybrid nanoparticles functionalized with sulfonic acid-terminated polyamidoamine | |
| CN110204749A (en) | Based on amination copolymerization microsphere and the perforated membrane and preparation method thereof for gathering more aldehyde | |
| Kaang et al. | Laminar flow-assisted synthesis of amorphous ZIF-8-based nano-motor with enhanced transmigration for photothermal cancer therapy | |
| EP1833855B1 (en) | Method for controlling the dimensions and the morphology of nanostructural polymeric materials, materials thereby obtained and uses thereof | |
| CN110194845B (en) | Preparation method and application of membrane flux-adjustable unsupported Janus monomolecular layer protein membrane material | |
| CN109223726A (en) | A kind of preparation method of Novel hollow capsule and obtained Novel hollow capsule | |
| CN112011041B (en) | Cyclodextrin monomolecular micelle and preparation method thereof | |
| CN110407967B (en) | Polymer with chiral characteristics, nano material thereof and synthetic method thereof | |
| CN113307922B (en) | Preparation method of polymer nano material with adjustable reduction response capability |
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 |