CN105078890B - A kind of preparation method of the multi-layer biological base vesica of releasable insulin - Google Patents
A kind of preparation method of the multi-layer biological base vesica of releasable insulin Download PDFInfo
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Abstract
A kind of preparation method of the multi-layer biological base vesica of releasable insulin, belongs to field of functional materials.The present invention modifies sodium alginate (Alg) and chitosan oligosaccharide (CS) synthesis Alg g CPBA graft copolymers and CS g CPBA graft copolymers respectively using biology base macromolecular as primary raw material, with 3 amino phenyl boric acid (APBA) of function monomer and 3 Carboxybenzeneboronic acids (CPBA)；Using the Nano particles of silicon dioxide for filling insulin in advance as template, then by LBL self-assembly, multi-layer biological base macromolecular microspheres are prepared；Add CaCl2, make Alg layers to be physical crosslinking, improve microballoon stability；Removing template is finally removed, prepares the biology base macromolecular vesica of controlled release insulin.Obtained vesica has high stability, unique hollow-core construction and excellent biocompatibility, and 60% is may be up to the covering amount of insulin；Under the stimulation of the glucose containing various concentration, insulin can realize controlled release, and burst size reaches 80%；This vesica has potential application value in fields such as drug loading, control releases.
Description
Technical field
A kind of preparation method of the multi-layer biological base vesica of releasable insulin, belongs to field of functional materials.
Background technology
Diabetes are to endanger one of major disease of human health, and traditional therapy brings huge pain to patient
Hardship, and intelligent sharp response medicine transmission polymer has the function of controllability and intelligentized control method drug delivery, can avoid
The shortcomings that traditional therapy, the treatment for diabetes provide new method.In these methods, phenyl boric acid and its derivative object
System stablizes, there is controllable adjustment insulin to deliver and persistently detect the application prospect of blood glucose and studied extensively.Phenyl boric acid
And its there are two different forms for derivative：It is unionized hydrophobic form when pH ＜ phenyl boric acid pKa values；When pH ＞ benzene
Boronic acid pKa, the hydrophilic version for ionization.The phenyl boric acid of hydrophilic version is very easy to and 1,2 glycol such as glucose after ionization
Deng generation reversible covalent bonds, more hydrophilic form is formed, and under extraneous glucose stimulation, competition is detached from matrix sugar
Class realizes that blood glucose is delivered and persistently detected to insulin with this.In existing research report, part has been selected in studying can not
Degradation or catabolite, which are harmful to organism, waits the poor polymer of biocompatibilities, so as to limit further should for they
With.
Sodium alginate (Sodium alginate, Alg) has hypotoxicity, good biocompatibility, thickening property, film forming
Property and gelling performance and height degradability, these special performances cause alginic acid and salt medical industry, textile and dyeing industry,
It is applied widely in the various fields such as fine chemical industry and food industry, and shows outstanding application value.It is early
United States Pharmacopeia has just been taken in 1938 and in 1963 annual income British Pharmacopoeias, and there is stability needed for pharmaceutical preparation auxiliary material, molten
Xie Xing, viscosity and safety.
Chitosan (chitosan, CS) can combine acid molecule, be natural polysaccharide because containing free amine group in its molecular structure
In unique alkaline polysaccharide so that chitosan have many special physicochemical properties and physiology, pharmacological function.As
Second largest organic natural resources in nature, chitosan itself have physiological adaptation, biocompatibility, can completely decomposability,
A variety of excellent properties such as polyfunctional reactant, stereochemical structure, recyclability and hydrophily so that the research of chitosan is got over application
To get over by worldwide extensive concern.
3- amino phenyl boric acid (APBA) and 3- Carboxybenzeneboronic acid (CPBA) of the present invention selection with sugared response are right respectively
Alg and CS carries out graft modification；Again using silica as template, fill insulin after, using after Alg deprotonations formed-
COO﹣Formation-NH after anion and CS protonations3 ﹢Electrostatic interaction between cation, is constructed by layer-by-layer
Nucleocapsid particle；Add CaCl2, make Alg layers to be physical crosslinking, improve microballoon stability；Finally use HF/NH4F is buffered
Liquid forms biology base macromolecular vesica after removing silica.It can shape between function monomer APBA and CPBA and macromolecular glucose
Into reversible ester bond, impart vesica glucose responding, by controlling the proportioning of macromolecular and function monomer, the grain size of vesica and
Structure, which can stimulate extraneous glucose, carries out the corresponding controlled release for realizing insulin of intelligence so that in drug loading, control
The fields such as release have broad application prospects.
Invention content
The present invention is directed to be prepared using silica as template using electrostatic interaction between the biological base electrolyte of functionalization
With the biology base vesica to insulin controlled release.
Technical scheme of the present invention：Sodium alginate (Alg) and chitosan oligosaccharide (CS) biology base macromolecular, selection have sugar response
Property 3- amino phenyl boric acid (APBA) and 3- Carboxybenzeneboronic acids (CPBA) graft modification is carried out to Alg and CS respectively；Again with dioxy
SiClx nano-particle is template, fills insulin, using the electrostatic interaction between Alg and CS, passes through layer-by-layer
Construct multi-layer core-shell structure microballoon；Add CaCl2, sodium alginate layer is physical crosslinking, improves microballoon stability；Finally
Use HF/NH4F buffer solutions form biology base macromolecular vesica after removing silica；And pass through and control macromolecular and function monomer
Proportioning, the grain size and structure of vesica, which are capable of to stimulate extraneous glucose, carries out the corresponding controlled release for realizing insulin of intelligence.
Using 3- amino phenyl boric acid as function monomer, by amidation process graft modification sodium alginate, sodium alginate and 3-
The mol ratio of amino phenyl boric acid is 10：The grafting rate of 1~8,3- amino phenyl boric acid is 10%~50%, gained graft polymers
Structure be：
It is expressed as Alg-g-APBA.
Using 3- Carboxybenzeneboronic acids as function monomer, by amidation process modified chitosan oligosaccharide, chitosan oligosaccharide and 3- carboxyl benzene boron
The mol ratio of acid is 10：The grafting rate of 1~5,3- Carboxybenzeneboronic acid is 5%~25%, and the structure of resulting polymers is：
It is expressed as CS-g-CPBA.
Using the Nano particles of silicon dioxide of 200nm as template, and with silane coupling agent 3- aminopropyl triethoxysilanes pair
Its surface carries out aminated；And then the Nano particles of silicon dioxide that surface amine groups are prepared is scattered in the buffering of pH=5
In solution, the insulin solutions of pH=5 are added in, and are added dropwise to the solution of the Alg-g-APBA and CS-g-CPBA of pH=5 in succession
In, it is repeated several times, assembling forms multi-layer core-shell structure microballoon；Finally, HF/NH is used4F buffer solutions remove silica template, obtain
To biology base macromolecular vesica.The assembling number of plies of Alg-g-APBA and CS-g-CPBA is 5~7 layers, and the covering amount of insulin is 60
~70%, the grain size of multi-layer core-shell structure microballoon is 240~270nm.
Beneficial effects of the present invention：The present invention respectively select with anion and cation biology base macromolecular Alg and
CS is polyelectrolyte, APBA the and CPBA function monomers and and macromolecular to glucose responding are respectively connected to by amidation process
Reversible ester bond can be formed between polysaccharide, imparts vesica glucose responding；Using Nano particles of silicon dioxide as template, fill out in advance
Insulin is buried, then by LBL self-assembly, prepares multi-layer biological base macromolecular microspheres；Add CaCl2, make Alg layers of generation
Physical crosslinking improves microballoon stability；Removing template is finally removed, prepares the biology base macromolecular vesica to insulin controlled release；
By controlling the proportioning of macromolecular and function monomer, the grain size and structure of vesica, which can stimulate extraneous glucose, carries out intelligent phase
It should realize the controlled release of insulin so that have broad application prospects in fields such as drug loading, control releases.
Description of the drawings
The infrared spectrum of Figure 1A lg-g-APBA
The nmr spectrum of Fig. 2 CS-g-CPBA
Fig. 3 SiO2Microballoon (a), ((Alg-g-APBA/CS-g-CPBA)5)@SiO2Core-shell particles (b) and ((Alg-g-
APBA/CS-g-CPBA)5) hollow vesica (c) transmission electron microscope picture
Fig. 4 ((Alg-g-APBA/CS-g-CPBA)5) hollow vesica insulin under different sugar concentration release profiles
Specific embodiment
The preparation of embodiment 1, CS-g-CPBA
1.790gCPBA (5mmol) is taken to be dissolved in 200ml deionized waters, it is sub- to add in co-catalyst 1.725gN- hydroxysuccinimidyls acyl
Amine (NHS, 15mmol) and 0.960g1- (3- dimethylamino-propyls) -3- ethyl carbodiimides (EDC, 5mmol), under magnetic agitation
Activate 0.5h.Then the chitosan oligosaccharide (5mmol) of the amount of substance such as CPBA is added in into solution, after reacting 72h at 25 DEG C, by sample
Dialysis one week, freeze-drying.
The preparation of embodiment 2, Alg-g-APBA
Alg (6mmol) is taken to be dissolved in 150ml water, adds in co-catalyst 2.070gNHS (18mmol) and 1.152g EDC
(6mmol) activates 0.5h under magnetic agitation.Then APBA (4mmol) is added in into solution, after reacting 72h at 25 DEG C, by sample
Product are dialysed 7 days, freeze-drying.
Embodiment 3, ((Alg-g-APBA/CS-g-CPBA)5)@SiO2Preparation
Alg-g-APBA, CS-g-CPBA, insulin are configured to the solution of a concentration of 2mg/ml of pH=5, SiO2-NH2Match
The suspension of a concentration of 2mg/ml of pH=5 is made, is ultrasonically treated 30min and is uniformly dispersed.Take the prepared SiO of 20ml2-NH2It suspends
Liquid is first slowly dropped in 20ml insulin solutions, reacts 30min, then 20mlAlg-g-APBA solution is slowly added dropwise, and is reacted
30min is centrifuged, and being washed with the aqueous solution of pH=5 will be in the aqueous solution of nano-particle redisperse to pH=5 after 3 times
(20ml) completes first layer assembling.First layer assembling hanging drop is added in 20mlCS-g-CPBA solution, after reacting 30min
It centrifuges, is washed (20ml) in the aqueous solution of nano-particle redisperse to pH=5 after 3 times, completed with the aqueous solution of pH=5
The second layer assembles.Third layer, the 4th layer, layer 5 assemble method reference first layer and the second layer.After layer 5 is completed, use
Cord blood after the aqueous solution centrifugation of pH=5 is washed 3 times.Every layer of eluent 1mL is taken, it is molten to add in 8mL Coomassie brilliant blues
Liquid (1mg/ml) fully vibrates, and absorbance at 595 is tested after 2min, and the covering amount of insulin is tested according to standard curve.
The preparation of embodiment 4, biology base macromolecular vesica
It takes ((Alg-g-APBA/CS-g-CPBA)5)@SiO2Sample 40mg adds in the NH of 100mlpH=54F/HF solution,
Ultrasonic disperse certain time, which is placed on, is stirred at room temperature etching 12h, and with deionized water centrifuge washing 3 times, vacuum drying obtains
((Alg-g-APBA/CS-g-CPBA)5) vesica.
The release in vitro research of embodiment 5, insulin
Take ((Alg-g-APBA/CS-g-CPBA) of appropriate load insulin5) vesica is configured to the suspension of 1mg/mL,
It is placed in dialysis (molecule in the PBS buffer solutions (pH=7.4) of different sugar concentration (0mg/mL, 2mg/mL, 6mg/mL, 20mg/mL)
Amount 8000~14000), be stirred at room temperature, 1h, 2h, 3h, 4h, 5h, 6h, 7h, 8h, 12h, for 24 hours, 36h take dialyzate 1mL, add in
8mL Coomassie Brillant Blue solutions (1mg/ml) fully vibrate, and absorbance at 595 is tested after 2min, and pancreas islet is tested according to standard curve
The burst size of element.
Claims (4)
1. a kind of preparation method of the biology base macromolecular vesica of releasable insulin, it is characterized in that few based on sodium alginate and shell
Sugared biology base macromolecular, it is few to sodium alginate and shell respectively with the 3- amino phenyl boric acid with sugared response and 3- Carboxybenzeneboronic acids
Sugar carries out graft modification, prepares sodium alginate-g- amino phenyl boric acid graft copolymers and the grafting of chitosan oligosaccharide-g- Carboxybenzeneboronic acids altogether
Polymers；Embed insulin in the Nano particles of silicon dioxide of surface amine groups, as template, and then using sodium alginate and
Electrostatic interaction between chitosan oligosaccharide constructs multi-layer core-shell structure microballoon by layer-by-layer；Add CaCl2, make
Sodium alginate layer is physical crosslinking, and improves microballoon stability；Finally, HF/NH is used4F buffer solutions remove silica template, obtain
To biology base macromolecular vesica；
Using 3- amino phenyl boric acid as function monomer, by amidation process graft modification sodium alginate, sodium alginate and 3- amino
The mol ratio of phenyl boric acid is 10：The grafting rate of 1~8,3- amino phenyl boric acid is 10%~50%, the knot of gained graft polymers
Structure is：
It is expressed as Alg-g-APBA；
Using 3- Carboxybenzeneboronic acids as function monomer, by amidation process modified chitosan oligosaccharide, chitosan oligosaccharide and 3- Carboxybenzeneboronic acids
Mol ratio is 10：The grafting rate of 1~5,3- Carboxybenzeneboronic acid is 5%~25%, and the structure of resulting polymers is：
It is expressed as CS-g-CPBA.
2. preparation method according to claim 1, it is characterized in that template is Nano particles of silicon dioxide, grain size is
200nm, and with silane coupling agent 3- aminopropyl triethoxysilanes its surface is carried out aminated；And then table will be prepared
The aminated Nano particles of silicon dioxide in face is scattered in the buffer solution of pH=5, adds in the insulin solutions of pH=5, pancreas islet
The covering amount of element is 60~70%.
3. preparation method according to claim 1, it is characterized in that the number of plies for the multi-layer vesicles that LBL self-assembly is formed for 5~
7 layers, the grain size of multi-layer core-shell structure microballoon is 240~270nm.
4. the biology base macromolecular vesica that preparation method according to claim 1 obtains.
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| CN106310230B (en) * | 2016-08-24 | 2019-08-27 | 华南师范大学 | A kind of oral insulin of LBL self-assembly structure transports the preparation method and application of system |
| CN106236734A (en) * | 2016-08-26 | 2016-12-21 | 郑州大学 | The preparation of mesoporous silicon oxide/insulin nanoparticles that phenylboric acid is modified and application |
| CN107320733A (en) * | 2017-07-01 | 2017-11-07 | 台州学院 | The preparation method of sugar response insulin carrier under a kind of physiological condition |
| CN108421042B (en) * | 2018-04-17 | 2021-02-09 | 南京邮电大学 | Preparation method of light-operated degradable hydrogel |
| CN108752488A (en) * | 2018-05-04 | 2018-11-06 | 暨南大学 | A kind of preparation method of carragheen grafting phenyl boric acid hydrophobic nanoparticles |
| CN113754793B (en) * | 2020-06-05 | 2023-12-15 | 中国医学科学院药物研究所 | Phenylboronic acid grafted chitosan oligosaccharide derivative and preparation method and application thereof |
| KR20230098561A (en) * | 2020-08-06 | 2023-07-04 | 푼다시오 유레카트 | Hybrid Core-Shell Microcapsules for Encapsulation of Active Ingredients |
| CN112618515B (en) * | 2020-12-29 | 2021-09-24 | 江南大学 | Preparation method of exosome-loaded oral colon-targeted drug delivery polymer |
| CN114099473B (en) * | 2021-11-30 | 2023-04-07 | 华臻 | Slow-release microcapsule for inducing in-vitro osteogenic differentiation of MC3T3-E1 cells and preparation method thereof |
| CN115025279B (en) * | 2022-06-30 | 2023-03-28 | 西安交通大学 | Sprayable natural hydrogel system for preventing postoperative adhesion and preparation and application thereof |
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Non-Patent Citations (3)
| Title |
|---|
| Layer-by-layer deposited nano- and micro-assemblies for insulin delivery: A review;Kentaro Yoshida等;《Materials Science and Engineering C》;20131005;第34卷;第384–392页 * |
| 层层自组装膜的研究： 从基础到生物医学领域中的应用;关英等;《高分子通报》;20130131(第1期);第40-52页 * |
| 糖响应性生物基聚电解质胶囊的制备与性能研究;冉茂双等;《化学学报》;20150721;第73卷;第1047-1054页 * |
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