CN106729638A - A kind of preparation method of poly-dopamine microcapsules oral insulin drug administration carrier - Google Patents
A kind of preparation method of poly-dopamine microcapsules oral insulin drug administration carrier Download PDFInfo
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Abstract
A kind of preparation method of poly-dopamine microcapsules oral insulin drug administration carrier, first by insulin powder dissolving in an acidic solution, adds sodium chloride, and insulin micro-nano granules are prepared by salting out method；Then insulin micro-nano granules are dispersed in the weakly alkaline solution of dopamine, the lower reaction of concussion obtains the Insulin Core shell structure of poly-dopamine cladding；Finally the Insulin Core shell structure that poly-dopamine is coated is dispersed in the solution of functional molecular, in its surface modification functional molecular, the poly-dopamine microcapsules oral insulin drug administration carrier of the biocompatibility for obtaining that gut barrier can be overcome, poly-dopamine microcapsules oral insulin drug administration carrier has the insulin releasing behavior of pH responses, good cell compatibility, the advantages of gut barrier being overcome, can apply to the treatment of diabetes.
Description
Technical field
The present invention relates to insulin administration carrier technique field, and in particular to a kind of poly-dopamine microcapsules oral insulin
The preparation method of drug administration carrier.
Background technology
Insulin is treatment the most frequently used and maximally effective medicine of diabetes.At present, the most frequently used administering mode of insulin is
Subcutaneous administrations.This administering mode easily causes the lipodystrophy of infection, allergic reaction and injection site, and
Injection pain, also bring very big puzzlement to patient the shortcomings of need special messenger to be administered.Therefore, research and development are needed badly more to pass through
Ji, convenient and painless novel insulin delivery system.
Oral insulin delivery system is a kind of novel insulin delivery system for developing in recent years, the physiology of this system
Interior raw path after mechanism of action is secreted in vivo to insulin is similar, i.e., insulin first passes around liver rather than directly arrival
Blood circulation system.Liver plays an important role in blood sugar level regulation, and it is also most important pancreatic secretion that it is first
The target organ of insulin.However, the insulin for there was only sub-fraction (about 20%) during using subcutaneous administrations is reached
Liver, a large amount of insulin are directly entered blood circulation system, easily trigger hypoglycemia.Oral insulin delivery system is simulated
The physiological mechanism of human glucose metabolism, insulin first passes around liver after being absorbed, reduce the insulin in system circulation
Level, so as to reduce the risk of hypoglycemic episodes.But, the realization of oral insulin delivery system still suffers from much asking
Topic.The oral administration of insulin is needed by the extreme environment in intestines and stomach, acid gastric juice and the intestinal juice of alkalescence and various
Digestive ferment, and it also requires passing through enteron aisle mucus barrier and epithelium layer barrier.Insulin is used as a kind of protein-based medicine
Thing, is easy to by protease hydrolytic in human gastrointestinal tract, loses biological function.This brings for the realization of Insulin Oral Delivery
Great challenge.
Interior in the past few decades, the fast development of nanosecond science and technology and Nano medication brings for oral bio molecules like pharmaceuticals
New development opportunity.Researchers have designed and developed multi-medicament carrier, such as polymer nano granules, liposome, micella, micro-
Capsule, microgel etc., coating decoration is carried out to insulin.These pharmaceutical carriers can protect the 26S Proteasome Structure and Function of insulin not by
Destruction, so as to improve the absorptivity of insulin.At present, the primitive of constructing of oral insulin carrier mainly includes that shitosan, Portugal gather
The natural polymer such as sugar, sodium alginate and hyaluronic acid, and the synthetic polymer such as PLA, polypropylene amine and polycaprolactone.
However, the preparation process of these pharmaceutical carriers usually needs to use the means such as high temperature, ultrasound, violent stirring, easily to pancreas islet
The structure and bioactivity of element are damaged.Therefore, scientific research personnel always searches for the preparation method of more preferable insulin carrier
And material.
In recent years, dopamine generates poly-dopamine due to oxidation auto polymerization reaction can occur under weak basic condition, and
And this auto polymerization reaction can occur on the surface of various inorganic and organic materials, and turn into a kind of novel preparation surface
The bionical of coating material constructs primitive.According to the literature, this extremely strong sticking property of poly-dopamine is because its molecular structure
Chemical constitution with the talin of aquatile mussel is close, and the two is rich in 3,4- dihydroxy-L-phenylalanine and lysine
Construction unit.Additionally, the phenolic hydroxyl group and amino group of poly-dopamine coating surface assign its chemical reactivity higher, such as phenol
Dehydration condensation, the complex reaction of phenolic hydroxyl group and many kinds of metal ions, the Michael's addition of amino group of hydroxyl and sulfydryl
With schiff base reaction etc., can be reacted with various chemical moleculars.Researchers are often using these reactions to poly- DOPA
Amine coating carries out surface modification, so as to prepare multi-functional composite.Poly-dopamine coating is except with extremely strong adhesive capacity
Outside chemical reactivity higher, also with very excellent biocompatibility, so that it has in biomedicine field
Very big application prospect.
Above-mentioned analysis result shows, it is difficult to prepare have good bio-compatible by gentle, quick and controllable reaction
Property, the oral insulin drug administration carrier of gastrointestinal tract barrier can be overcome, bionical poly-dopamine material is insulin delivery system
Preparation brings new opportunity to develop.
The content of the invention
It is oral it is an object of the invention to provide a kind of poly-dopamine microcapsules in order to overcome the shortcoming of above-mentioned prior art
The preparation method of insulin administration carrier, the carrier has good biocompatibility, gastrointestinal tract barrier can be overcome, with pancreas
The controllable advantage of island element rate of release.
In order to achieve the above object, the technical scheme taken of the present invention is：
A kind of preparation method of poly-dopamine microcapsules oral insulin drug administration carrier, comprises the following steps：
1) concentration is reached 5-50mg/mL in an acidic solution insulin powder dissolving, be added thereto to sodium chloride, make
The content of sodium chloride is 0.5-5mol/L, and insulin micro-nano granules are prepared by salting out method；
2) insulin micro-nano granules are dispersed in the weakly alkaline solution of dopamine so that the concentration of insulin is 10-
80mg/mL, the concentration of dopamine is 0.5-5mg/mL, and the lower reaction of concussion obtains the Insulin Core shell structure of poly-dopamine cladding；
3) the Insulin Core shell structure that poly-dopamine is coated is dispersed in the solution of functional molecular so that poly-dopamine
The concentration of the Insulin Core shell structure of cladding is 10-80mg/mL, and the concentration of functional molecular is 5-10mg/mL, in core shell structure
Surface modification functional molecular, the poly-dopamine microcapsules oral insulin of the biocompatibility for obtaining that gut barrier can be overcome
Drug administration carrier.
Described insulin be actrapid monotard, bovine insulin, pork insulin, sheep insulin, horse insulin, dog insulin or
One or more in cat insulin.
Described acid solution is hydrochloric acid solution, and its pH is 1-2.
The described water-bath temperature for preparing insulin micro-nano granules by salting out method is 5-20 DEG C；Reaction condition is
Shake, stir or ultrasonically treated；Reaction time is 0.5-3h.
Described salting out method prepares insulin micro-nano granules, by being dispersed in 2mol/L sodium chloride solutions after preparation
Row cleaning, centrifugation obtains pure insulin micro-nano granules.
Described weakly alkaline solution is Tris-HCl solution, Tris-HCl and H2O2Solution or NaOH and H2O2Solution,
The pH of solution is 7.5-10.5.
The concussion rotating speed reacted under described concussion is 500-1500rpm；Reaction time is 12-48h；Reaction temperature is 4-
30℃。
The Insulin Core shell structure of described poly-dopamine cladding is cleaned by being centrifuged, being dispersed in weakly alkaline solution, and
It is stored in 4 DEG C of environment.
Described functional molecular is stronger interaction can occur with hydrophobic, negatively charged enteron aisle mucus
Or the biological targeting molecule of specific recognition can be carried out with the glycoprotein in enteron aisle mucus, including polyethylene glycol, shitosan,
Cationic-liposome and agglutinin, these molecules can improve carrier stability in the gastrointestinal tract, extend it in enteron aisle mucus
The interior residence time, and then improve the absorptivity of insulin.
Described step 3) by covalent bond or non-covalent interaction in poly-dopamine surface of microcapsule rhetorical function
Property molecule.
Described step 3) the specific reaction of functional molecular is in poly-dopamine by the schiff base reaction with glutaraldehyde
Surface of microcapsule modification shitosan, cationic-liposome and agglutinin, are modified by the dehydration condensation of phenolic hydroxyl group and sulfydryl
Peg molecule with sulfydryl.
The application of above-mentioned poly-dopamine microcapsules oral insulin drug administration carrier, including test poly-dopamine microcapsules pancreas islet
The interaction of plain drug administration carrier and mucoprotein, the carrier of evaluation function sex modification overcomes the ability of gut barrier；Test is poly- more
The insulin releasing behavior of bar amine microcapsules insulin administration carrier；Test the thin of poly-dopamine microcapsules insulin administration carrier
Born of the same parents' compatibility, poly-dopamine microcapsules insulin administration carrier is used to prepare the product of the following diseases for the treatment of：Human diabetes, ox
Diabetes, pig diabetes, sheep diabetes, horse diabetes, dog diabetes and cat diabetes.
Beneficial effects of the present invention are：
1) gentle, the quick and controllable reaction of poly-dopamine microcapsules oral insulin drug administration carrier first passage of the present invention
Prepare with good biocompatibility, can overcome the bionical poly-dopamine microcapsules oral insulin of gut barrier that load is administered
Body.
2) poly-dopamine microcapsules oral insulin drug administration carrier of the present invention can be by changing insulin in course of reaction
Concentration, the concentration of sodium chloride when saltouing, the concentration of dopamine, the type of cushioning liquid, pH value, and the reaction time, temperature,
The reaction conditions such as concussion speed, realize the regulation and control to poly-dopamine microcapsules wall thickness and size, so as to regulate and control carrying medicament pancreas islet
The rate of release of element.
3) poly-dopamine microcapsules oral insulin drug administration carrier good biocompatibility of the present invention, can overcome enteron aisle screen
Barrier, the residence time in enteron aisle mucus is long, so as to improve the absorptivity of insulin, improves the operational efficiency of delivery system.
Brief description of the drawings
Fig. 1 be (a) prepared by embodiment 1 it is uncoated, (b) coated the insulin granule of poly-dopamine microcapsules shell
Stereoscan photograph.
Fig. 2 is the insulin releasing curve of chitosan-modified oral insulin drug administration carrier prepared by embodiment 1：(a)
The PBS cushioning liquid of pH 7.4, the PBS cushioning liquid of (b) pH 5.4.
Fig. 3 is the cytotoxicity test of chitosan-modified oral insulin drug administration carrier prepared by embodiment 1, and (a) is
Control sample, (b), (c) are respectively human umblilical vein endothelial after the chitosan-modified oral insulin drug administration carrier of the μ L of addition 20,40
The survival rate of cell.
Specific embodiment
With reference to embodiment, the present invention will be described in detail.Unless it is defined otherwise in the description of the present invention, otherwise
Herein all of technical term be all according to persons skilled in the art usually using and the conventional definitions that understand use.
Experimental technique described in following embodiments, unless otherwise specified, is conventional method；The reagent and material, such as without special theory
It is bright, commercially obtain.
A kind of embodiment 1, preparation method of poly-dopamine microcapsules oral insulin drug administration carrier, comprises the following steps：
1) insulin powder is dissolved using the hydrochloric acid solution that pH is 2, makes its concentration for 10mg/mL, be added thereto to chlorination
Sodium crystal, lower reaction 1h is shaken until the amount of sodium chloride reaches 0.6mol/L, in 15 DEG C of water-baths, it was observed that the insulin of white
Grain is gradually separated out, centrifugation product, and uses 2mol/L sodium chloride solutions cleaning product 2 times, and centrifugation obtains insulin
Micro-nano granules；
2) the Tris-HCl cushioning liquid of pH 7.5 first, is prepared；Secondly, insulin micro-nano granules are scattered in
Tris-HCl cushioning liquid so that the concentration of insulin is 10mg/mL, adds dopamine powder, makes its concentration for 2mg/mL, will
Above reaction system is placed at room temperature with the rotating speed concussion reaction 24h of 900rpm, and in course of reaction, the color of solution is by pink
Color, is gradually converted into brown, dark-brown；Finally, particle is collected by centrifugation, particle is cleaned using Tris-HCl cushioning liquid, obtain
The Insulin Core shell structure of poly-dopamine cladding, is stored in 4 DEG C of environment；Its microstructure is as shown in figure 1, Fig. 1 a display brand-news
Insulin granule show slightly coarse, and after poly-dopamine shell has been coated, insulin becomes very smooth, as shown in Figure 1 b；
3) the Insulin Core shell structure that first, poly-dopamine is coated, is distributed in the PBS solution of 0.025% glutaraldehyde
(pH 7.4) so that the concentration of the Insulin Core shell structure of poly-dopamine cladding is 10mg/mL, is adsorbed 24 hours；Washing three times
Afterwards, the PBS solution of 5mg/mL shitosans is added, is adsorbed 12 hours, washed three times；Centrifugation, obtains chitosan-modified mouth
Insulin administration carrier is taken, is stored in 4 DEG C of environment.
The beneficial effect of the chitosan-modified oral insulin drug administration carrier of the present embodiment：
(1) mucoprotein adsorption experiment：
Prepare the mucoprotein solution of 0.5mg/mL, pH 5.5.The chitosan-modified oral insulin of above-mentioned preparation is given
Drug carrier is dispersed in mucoprotein solution, and interact 5h in 37 DEG C of environment.Centrifugation, mucoprotein contains in measurement supernatant
Amount, so as to calculate the quality that drug administration carrier adsorbs mucoprotein, evaluates the power of drug administration carrier and mucus effect.
(2) insulin releasing performance testing：
Two kinds of PBS cushioning liquid that pH 7.4 and pH 5.4 is respectively adopted study the insulin of oral insulin drug administration carrier
Release behavior.Oral insulin drug administration carrier is scattered in (2mL) in two kinds of cushioning liquid respectively, is shaken under conditions of 200rpm
Swing uelralante.The testing time is 24h altogether.At specific time point, the supernatant of 0.5mL is taken out in centrifugation, and replaces with new
Cushioning liquid processed.By the insulin concentration in Coomassie brilliant blue solution testing supernatant, so as to show that insulin is slow at two kinds
The release profiles rushed in solution.The insulin releasing performance of oral insulin drug administration carrier is as shown in Figure 2.Fig. 2 explanations, oral pancreas
The insulin releasing of island element drug administration carrier is presented pH responses, and in the cushioning liquid of pH 5.4, the release of insulin is little.And
In the cushioning liquid of pH 7.4, insulin realizes complete release.The release behavior of this response, is conducive to the carrier to lead to
Oral mode is crossed to be administered.
(3) cell compatibility of oral insulin drug administration carrier：
Cytotoxicity test is carried out by taking Human umbilical vein endothelial cells (HUVEC) as an example.HUVEC cell-seeding-densities are about
2×104cells/mL.Cell is cultivated in polystyrene culture dish.Hyclone containing 10% (v/v%), 1% in culture medium
Streptomysin and penicillin, every two days of culture medium changes once.Incubator temperature is set as 37 DEG C, contains 5% (v/v%) CO2
Humid air.When cell reaches for 90% growth period, they are divided with the digestion techniques of standard is trained in 24 orifice plates
Support.Culture situation according to cell, about two days later, draws nutrient solution, renews liquid, wherein being added without any sample in a row, makees
It is reference, is separately added into different volumes (20,40 μ L) oral insulin drug administration carrier solution in several row in addition, continues to cultivate 12
Hour.It is each in every hole to add 100 μ L MTT solution (concentration is 5mg/mL) in order to determine the activity of cell, wrapped up with aluminium foil
Culture plate incubates 4h in incubator.Culture medium and MTT in removal hole, are then separately added into 200 μ L DMSO in each hole,
Concussion 10min, MTT- formazan crystal of the abundant dissolution precipitation in culture plate bottom.Detected using ultraviolet-visible spectrometer immediately
Its light absorption value at 570nm, to prevent product deterioration.Result is as shown in Figure 3.From the figure 3, it may be seen that the addition of microcapsules is not right
The growth of cell is impacted, and illustrates microcapsules no cytotoxicity, is bio-compatible.
A kind of embodiment 2, preparation method of poly-dopamine microcapsules oral insulin drug administration carrier, comprises the following steps：
1) insulin powder is dissolved using the hydrochloric acid solution that pH is 1, makes its concentration be 50mg/mL；It is added thereto to chlorination
Sodium crystal, until the amount of sodium chloride reaches 0.5mol/L；Lower reaction 3h is shaken in 5 DEG C of water-baths, it was observed that the insulin of white
Grain is gradually separated out；Centrifugation, cleaning 2 times is carried out using 2mol/L sodium chloride solutions, and centrifugation obtains insulin micro-nano
Particle；
2) the Tris-HCl cushioning liquid of pH 7.5 is prepared, it is molten that insulin micro-nano granules are scattered in into Tris-HCl bufferings
Liquid so that the concentration of insulin is 80mg/mL, adds dopamine powder, makes its concentration be 2mg/mL；Above reaction system is put
At room temperature with the rotating speed concussion reaction 24h of 900rpm, in course of reaction, the color of solution is gradually converted into palm fibre by pink
Color, dark-brown；Particle is collected by centrifugation, particle is cleaned using Tris-HCl cushioning liquid, obtain the insulin of poly-dopamine cladding
Core shell structure, is stored in 4 DEG C of environment；
3) the Insulin Core shell structure that poly-dopamine is coated is distributed in the PBS of 0.025% glutaraldehyde
(PBS pH 7.4) so that the concentration of the Insulin Core shell structure of poly-dopamine cladding is 80mg/mL, is adsorbed 24 hours；Washing
After three times, the PBS solution of 10mg/mL shitosans is added, adsorbed 12 hours, washed three times；Centrifugation, obtains chitosan-modified
Oral insulin drug administration carrier, be stored in 4 DEG C of environment.
A kind of embodiment 3, preparation method of poly-dopamine microcapsules oral insulin drug administration carrier, comprises the following steps：
1) insulin powder is dissolved using the hydrochloric acid solution that pH is 1.5, makes its concentration be 20mg/mL；It is added thereto to chlorine
Change sodium crystal, until the amount of sodium chloride reaches 2mol/L；Lower reaction 1h is shaken in 15 DEG C of water-baths, it was observed that the insulin of white
Grain is gradually separated out；Centrifugation, cleaning 2 times is carried out using 2mol/L sodium chloride solutions, and centrifugation obtains insulin micro-nano
Particle；
2) the Tris-HCl cushioning liquid of pH 8.5 is prepared, upper insulin micro-nano granules is scattered in Tris-HCl bufferings
Solution so that the concentration of insulin is 40mg/mL, adds dopamine powder, makes its concentration be 2mg/mL；By above reaction system
It is placed at room temperature with the rotating speed concussion reaction 24h of 900rpm；In course of reaction, the color of solution is gradually converted into by pink
Brown, dark-brown.Particle is collected by centrifugation, particle is cleaned using Tris-HCl cushioning liquid, obtain the pancreas islet of poly-dopamine cladding
Plain core shell structure, is stored in 4 DEG C of environment；
3) the Insulin Core shell structure that poly-dopamine is coated is distributed in the PBS of 0.025% glutaraldehyde
(PBS pH 7.4) so that the concentration of the Insulin Core shell structure of poly-dopamine cladding is 40mg/mL, is adsorbed 24 hours；Washing
After three times, the PBS solution of 10mg/mL shitosans is added, adsorbed 12 hours, washed three times；Centrifugation, obtains chitosan-modified
Oral insulin drug administration carrier, be stored in 4 DEG C of environment.
A kind of embodiment 4, preparation method of poly-dopamine microcapsules oral insulin drug administration carrier, comprises the following steps：
1) insulin powder is dissolved using the hydrochloric acid solution that pH is 2, makes its concentration be 0.5mg/mL；It is added thereto to chlorination
Sodium crystal, until the amount of sodium chloride reaches 5mol/L；Lower reaction 1h is shaken in 10 DEG C of water-baths, it was observed that the insulin granule of white
Gradually separate out；Centrifugation, cleaning 2 times is carried out using 2mol/L sodium chloride solutions, and centrifugation obtains micro-nano of insulin
Grain；
2) the Tris-HCl cushioning liquid of pH 10 is prepared, upper insulin micro-nano granules is scattered in Tris-HCl bufferings
Solution so that the concentration of insulin is 30mg/mL, adds dopamine powder, makes its concentration be 2mg/mL；By above reaction system
It is placed at room temperature with the rotating speed concussion reaction 12h of 900rpm, in course of reaction, the color of solution is gradually converted into by pink
Brown, dark-brown.Particle is collected by centrifugation, particle is cleaned using Tris-HCl cushioning liquid, obtain the pancreas islet of poly-dopamine cladding
Plain core shell structure, is stored in 4 DEG C of environment；
3) first, the Insulin Core shell structure that poly-dopamine is coated is distributed to the phosphate-buffered of 0.025% glutaraldehyde
In solution (PBS pH 7.4) so that the concentration of the Insulin Core shell structure of poly-dopamine cladding is 30mg/mL, absorption 24 is small
When；After washing three times, the PBS solution of 8mg/mL shitosans is added, adsorbed 12 hours, washed three times；Centrifugation, obtains shell and gathers
Sugar-modified oral insulin drug administration carrier, is stored in 4 DEG C of environment.
A kind of embodiment 5, preparation method of poly-dopamine microcapsules oral insulin drug administration carrier, comprises the following steps：
1) insulin powder is dissolved using the hydrochloric acid solution that pH is 1.5, makes its concentration be 20mg/mL；It is added thereto to chlorine
Change sodium crystal, until the amount of sodium chloride reaches 2mol/L；Lower reaction 1h is shaken in 15 DEG C of water-baths, it was observed that the insulin of white
Grain is gradually separated out, centrifugation, and cleaning 2 times is carried out using 2mol/L sodium chloride solutions, and centrifugation obtains insulin micro-nano
Particle；
2) NaOH and H of pH 8.5 are prepared2O2Insulin micro-nano granules are scattered in NaOH and H by mixed solution2O2It is mixed
Close solution so that the concentration of insulin is 50mg/mL, adds dopamine powder, make its concentration be 0.5mg/mL；Reacted by more than
System is placed at room temperature with the rotating speed concussion reaction 24h of 900rpm.In course of reaction, the color of solution is gradually turned by pink
It is changed into brown, dark-brown, particle is collected by centrifugation, particle is cleaned using PBS cushioning liquid, obtains the insulin of poly-dopamine cladding
Core shell structure, is stored in 4 DEG C of environment；
3) the Insulin Core shell structure that poly-dopamine is coated is distributed in the PBS of 0.025% glutaraldehyde
(PBS pH 7.4) so that the concentration of the Insulin Core shell structure of poly-dopamine cladding is 50mg/mL, is adsorbed 24 hours；Washing
After three times, the PBS solution of 8mg/mL shitosans is added, adsorbed 12 hours, washed three times；Centrifugation, obtains chitosan-modified
Oral insulin drug administration carrier, be stored in 4 DEG C of environment.
A kind of embodiment 6, preparation method of poly-dopamine microcapsules oral insulin drug administration carrier, comprises the following steps：
1) insulin powder is dissolved using the hydrochloric acid solution that pH is 1.5, makes its concentration be 20mg/mL；It is added thereto to chlorine
Change sodium crystal, until the amount of sodium chloride reaches 2mol/L；Lower reaction 1h is shaken in 15 DEG C of water-baths, it was observed that the insulin of white
Grain is gradually separated out；Centrifugation, cleaning 2 times is carried out using 2mol/L sodium chloride solutions, and centrifugation obtains insulin micro-nano
Particle；
2) the Tris-HCl cushioning liquid of pH 10.5 is prepared, insulin micro-nano granules is scattered in Tris-HCl bufferings
Solution so that the concentration of insulin is 50mg/mL, adds dopamine powder, makes its concentration be 5mg/mL；By above reaction system
It is placed at room temperature with the rotating speed concussion reaction 24h of 900rpm, in course of reaction, the color of solution is gradually converted into by pink
Brown, dark-brown, are collected by centrifugation particle, and particle is cleaned using Tris-HCl cushioning liquid, obtain the pancreas islet of poly-dopamine cladding
Plain core shell structure, is stored in 4 DEG C of environment；
3) the Insulin Core shell structure that poly-dopamine is coated is distributed in the PBS of 0.025% glutaraldehyde
(PBS pH 7.4) so that the concentration of the Insulin Core shell structure of poly-dopamine cladding is 50mg/mL, is adsorbed 24 hours；Washing
After three times, the PBS solution of 10mg/mL agglutinins is added, adsorbed 12 hours, washed three times；Centrifugation, obtains Phytoagglutinin modified
Oral insulin drug administration carrier, be stored in 4 DEG C of environment.
According to the method in embodiment 1, using mucoprotein adsorption experiment, insulin releasing performance testing, MTT cytoactives
Experiment, the application performance of the oral insulin drug administration carrier for being prepared to embodiment 2-6 respectively is estimated.Result shows, implements
Oral insulin drug administration carrier prepared by example 2-6 shows and has a stronger interaction with mucoprotein, and carrier cell phase
Capacitive very well, with regulatable, pH responses insulin releasing behavior, is expected to be applied to the diabetes of human and animal
Treatment.
Claims (10)
1. a kind of preparation method of poly-dopamine microcapsules oral insulin drug administration carrier, it is characterised in that comprise the following steps：
1) concentration is reached 5-50mg/mL in an acidic solution insulin powder dissolving, be added thereto to sodium chloride, make chlorination
The content of sodium is 0.5-5mol/L, and insulin micro-nano granules are prepared by salting out method；
2) insulin micro-nano granules are dispersed in the weakly alkaline solution of dopamine so that the concentration of insulin is 10-80mg/
ML, the concentration of dopamine is 0.5-5mg/mL, and the lower reaction of concussion obtains the Insulin Core shell structure of poly-dopamine cladding；
3) the Insulin Core shell structure that poly-dopamine is coated is dispersed in the solution of functional molecular so that poly-dopamine is coated
The concentration of Insulin Core shell structure be 10-80mg/mL, the concentration of functional molecular is 5-10mg/mL, on core shell structure surface
Rhetorical function molecule, the poly-dopamine microcapsules oral insulin administration of the biocompatibility for obtaining that gut barrier can be overcome
Carrier.
2. the preparation method of a kind of poly-dopamine microcapsules oral insulin drug administration carrier according to claim 1, it is special
Levy and be：Described insulin be actrapid monotard, bovine insulin, pork insulin, sheep insulin, horse insulin, dog insulin or
One or more in cat insulin.
3. the preparation method of a kind of poly-dopamine microcapsules oral insulin drug administration carrier according to claim 1, it is special
Levy and be：Described acid solution is hydrochloric acid solution, and its pH is 1-2.
4. the preparation method of a kind of poly-dopamine microcapsules oral insulin drug administration carrier according to claim 1, it is special
Levy and be：The described water-bath temperature for preparing insulin micro-nano granules by salting out method is 5-20 DEG C；Reaction condition is
Shake, stir or ultrasonically treated；Reaction time is 0.5-3h.
5. the preparation method of a kind of poly-dopamine microcapsules oral insulin drug administration carrier according to claim 1, it is special
Levy and be：Described salting out method prepares insulin micro-nano granules, by being dispersed in 2mol/L sodium chloride solutions after preparation
Row cleaning, centrifugation obtains pure insulin micro-nano granules.
6. the preparation method of a kind of poly-dopamine microcapsules oral insulin drug administration carrier according to claim 1, it is special
Levy and be：Described weakly alkaline solution is Tris-HCl solution, Tris-HCl and H2O2Solution or NaOH and H2O2Solution, it is molten
The pH of liquid is 7.5-10.5.
7. the preparation method of a kind of poly-dopamine microcapsules oral insulin drug administration carrier according to claim 1, it is special
Levy and be：The concussion rotating speed reacted under described concussion is 500-1500rpm；Reaction time is 12-48h；Reaction temperature is 4-
30℃。
8. the preparation method of a kind of poly-dopamine microcapsules oral insulin drug administration carrier according to claim 1, it is special
Levy and be：The Insulin Core shell structure of described poly-dopamine cladding is cleaned by being centrifuged, being dispersed in weakly alkaline solution, and is protected
In the presence of in 4 DEG C of environment.
9. the preparation method of a kind of poly-dopamine microcapsules oral insulin drug administration carrier according to claim 1, it is special
Levy and be：Described functional molecular is stronger interaction can occur with hydrophobic, negatively charged enteron aisle mucus
Or the biological targeting molecule of specific recognition can be carried out with the glycoprotein in enteron aisle mucus, including polyethylene glycol, shitosan,
Cationic-liposome and agglutinin.
10. the preparation method of a kind of poly-dopamine microcapsules oral insulin drug administration carrier according to claim 1, it is special
Levy and be, comprise the following steps：
1) insulin powder is dissolved using the hydrochloric acid solution that pH is 2, makes its concentration for 10mg/mL, be added thereto to sodium chloride brilliant
Body, lower reaction 1h is shaken until the amount of sodium chloride reaches 0.6mol/L, in 15 DEG C of water-baths, it was observed that the insulin granule of white by
Gradually separate out, centrifugation product, and use 2mol/L sodium chloride solutions cleaning product 2 times, centrifugation obtains insulin micro-nano
Rice grain；
2) the Tris-HCl cushioning liquid of pH 7.5 first, is prepared；Secondly, insulin micro-nano granules are scattered in Tris-HCl
Cushioning liquid so that the concentration of insulin is 10mg/mL, adds dopamine powder, makes its concentration for 2mg/mL, is reacted by more than
System is placed at room temperature with the rotating speed concussion reaction 24h of 900rpm, and in course of reaction, the color of solution is gradually turned by pink
It is changed into brown, dark-brown；Finally, particle is collected by centrifugation, particle is cleaned using Tris-HCl cushioning liquid, obtain poly-dopamine bag
The Insulin Core shell structure for covering, is stored in 4 DEG C of environment；
3) the Insulin Core shell structure that first, poly-dopamine is coated, is distributed to (pH in the PBS solution of 0.025% glutaraldehyde
7.4) so that the concentration of the Insulin Core shell structure of poly-dopamine cladding is 10mg/mL, is adsorbed 24 hours；After washing three times, plus
Enter the PBS solution of 5mg/mL shitosans, adsorb 12 hours, wash three times；Centrifugation, obtains chitosan-modified oral pancreas islet
Plain drug administration carrier, is stored in 4 DEG C of environment.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107904190A (en) * | 2017-12-19 | 2018-04-13 | 南京工业大学 | A kind of method for the modification of cell surface functional coating |
| CN107897869A (en) * | 2017-11-13 | 2018-04-13 | 常州凯恒纺织品有限公司 | A kind of preparation method of ginger oil nanoemulsions |
| CN110179997A (en) * | 2018-08-08 | 2019-08-30 | 昆山新蕴达生物科技有限公司北京分公司 | A kind of nano-medicament carrier for treating diabetes and combinations thereof drug |
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| CN115463108A (en) * | 2022-09-16 | 2022-12-13 | 唐颐惠康干细胞产业平台(天津)有限公司 | Probiotics microcapsule for treating genital tract infection and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105412935A (en) * | 2015-02-04 | 2016-03-23 | 四川大学 | N-(2-hydroxypropyl) methacrylamide polymer based nanoparticles and preparation method thereof |
| CN106038478A (en) * | 2016-06-24 | 2016-10-26 | 天津大学 | Glucose sensitive porous microsphere/polymer composite gel as well as preparation method and application thereof |
| KR101830192B1 (en) * | 2015-06-25 | 2018-02-21 | 고려대학교 산학협력단 | Biocompatible fiber aggregate with expanded inter-fiber space and the method for manufacturing thereof |
-
2016
- 2016-12-14 CN CN201611154834.3A patent/CN106729638A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105412935A (en) * | 2015-02-04 | 2016-03-23 | 四川大学 | N-(2-hydroxypropyl) methacrylamide polymer based nanoparticles and preparation method thereof |
| KR101830192B1 (en) * | 2015-06-25 | 2018-02-21 | 고려대학교 산학협력단 | Biocompatible fiber aggregate with expanded inter-fiber space and the method for manufacturing thereof |
| CN106038478A (en) * | 2016-06-24 | 2016-10-26 | 天津大学 | Glucose sensitive porous microsphere/polymer composite gel as well as preparation method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| HONG LI等: "Facile fabrication of robust polydopamine microcapsules for insulin delivery", 《JOURNAL OF COLLOID AND INTERFACE SCIENCE》 * |
| 赵应征: "《生物药物制剂学》", 30 June 2011, 浙江大学出版社 * |
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| CN107904190A (en) * | 2017-12-19 | 2018-04-13 | 南京工业大学 | A kind of method for the modification of cell surface functional coating |
| CN107904190B (en) * | 2017-12-19 | 2021-10-15 | 南京工业大学 | Method for modifying functional coating on cell surface |
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| CN110200945A (en) * | 2019-07-18 | 2019-09-06 | 西安石油大学 | A kind of preparation method of light sensitivity DOPA amido nano-medicament carrier |
| CN111592801A (en) * | 2020-06-04 | 2020-08-28 | 广东省生物工程研究所（广州甘蔗糖业研究所） | Sustained and controlled release coating, sustained and controlled release mulching film and preparation method thereof |
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