CN106038478A - Glucose sensitive porous microsphere/polymer composite gel as well as preparation method and application thereof - Google Patents
Glucose sensitive porous microsphere/polymer composite gel as well as preparation method and application thereof Download PDFInfo
- Publication number
- CN106038478A CN106038478A CN201610493506.XA CN201610493506A CN106038478A CN 106038478 A CN106038478 A CN 106038478A CN 201610493506 A CN201610493506 A CN 201610493506A CN 106038478 A CN106038478 A CN 106038478A
- Authority
- CN
- China
- Prior art keywords
- porous microsphere
- plga
- acid
- microsphere
- polymer
- 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.)
- Granted
Links
Classifications
-
- 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
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/28—Insulins
-
- 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
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
-
- 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
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
-
- 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
- A61K9/0002—Galenical forms characterised by the drug release technique; Application systems commanded by energy
-
- 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
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/5005—Wall or coating material
- A61K9/5015—Organic compounds, e.g. fats, sugars
-
- 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
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/5005—Wall or coating material
- A61K9/5021—Organic macromolecular compounds
- A61K9/5031—Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poly(lactide-co-glycolide)
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Diabetes (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Endocrinology (AREA)
- Inorganic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to glucose sensitive porous microsphere/polymer composite gel as well as a preparation method and application thereof. In the glucose sensitive porous microsphere/polymer composite gel, porous microspheres are polylactic acid-glycolic acid copolymer (PLGA) porous microspheres, and the average particle size of the PLGA porous microspheres is 1-20mu m; and the polymer is hyaluronic acid containing a catechol group. The preparation method of the porous microsphere/polyhydroxylated polymer composite gel comprises the following steps: blending the porous microspheres subjected to surface modification by phenylboronic acid with the aqueous solution of the polyhydroxylated polymer; and carrying out bonding reaction between phenylboronic acid on the surfaces of the microspheres and cis-dihydroxy on the chain segment of the polyhydroxylated polymer to form a cross-linked polymer network, thereby obtaining the composite gel. The glucose sensitive porous microsphere/polymer composite gel is applied to a drug delivery system, is used for developing insulin implanted gel long-acting sustained release preparations, and has great application values in the field of development of a new dosage form for treating diabetes.
Description
Technical field
The present invention relates to a kind of porous microsphere/polymer pluralgel and preparation method thereof, this pluralgel system has
Glucose-sensitive, it is possible to the most a large amount of load insulins the change of response glucose concentration under the physiological condition of pH 7.4
The release of regulation insulin, belongs to medicinal macromolecule hydrogel field.
Background technology
Diabetes and complication thereof have become as the great public health crisis of one, China.Diabetes are insulin secretion phases
To or absolute metabolic disease caused by deficiency, so since finding from insulin and successfully extract, it is always diabetes and controls
The choice drug treated.At present, the main path that clinical islet element is administered is subcutaneous injection.Diabetics generally requires long-term frequency
Numerous injection, not only cost is high, be administered inconvenience, there is also many untoward reaction such as pain and subcutaneous tissue atrophy, thus patient
Compliance extreme difference.Additionally, this Therapeutic Method cannot provide a kind of simulation physiological insulin secretion pattern, it is impossible to effectively stop phase
Close generation [Pillai O., Panchagnula R.Insulin therapies:past, the present and of complication
future,Drug Discov.Today 2001,6(20),1056-1061].Therefore, safety convenient is provided for diabetics
Non-injection formula drug-delivery preparation there is important social meaning and wide economic outlook.
Implant the insulin physiology self-adjustable drug-supplying system of glucose-sensitive in subcutaneous or abdominal cavity, not only can avoid gastrointestinal
Road and the first pass effect of liver and biomembrane obstacle, improve bioavailability, it is achieved long-acting slow-release, it is to avoid frequent drug administration, increases
Patient compliance；But also can the change of automatic sensing blood sugar concentration in the patient and regulate and control the release of insulin in time,
The secretion pattern of big degree simulation physiological insulin, is ideal scheme [the Catargi B.Current for the treatment of diabetes
status and future of implantable insulin pumps for the treatment of diabetes,
Expert Rev.Med.Dev.2004,1,181-185.].Phenylboric acid and derivant thereof can reversible, persistently respond Fructus Vitis viniferae for a long time
Sugar concentration change, it is achieved the intelligence release of circulating insulin on-off formula (Closed-loop delivery).Therefore, based on benzene
The insulin implanted gel drug-supplying system of boric acid functionalized polymer exploitation, has implantation locating features and the benzene boron of gel simultaneously
The acid sensing identification ability to glucose, is not only able to by the way of Wicresoft or injection be easy to implant, and can
Simulation physiological insulin secretion pattern, bio-compatibility is good, lives patient without impact, can degradation in vivo after drug release is complete
Without taking out.Obviously, this kind of high-molecular gel has high at exploitation insulin physiology self-adjustable drug delivery implant system aspects
Using value.While it is true, polyalcohol hydrogel based on phenylboric acid could stimulate by response glucose the most in the basic conditions,
Glucose-sensitive function cannot be played under the physiological condition at pH 7.4, limit its application in vivo greatly
[Bratlie K.M.,York R.L.,Invernale M.A.,Langer R.,Anderson D.G.Materials for
diabetes therapeutics,Adv.Healthcare Mater.2012,1,267-284.].Additionally, this life of insulin
Thing macromolecular drug (Mn=5808g/mol), it is different from other small-molecule drugs, insulin is encapsulated completely, efficiently into solidifying
Extremely difficult [Vermonden T., Censi R., the Hennink W.E.Hydrogels for protein of glue carrier
delivery,Chem.Rev.2012,112,2853-2888].Additionally, there is the insulin of macromolecular structure, and macromolecule water
Often there is stronger intermolecular force between gel, be loaded into a large amount of insulin to the mechanical property of gel, solvability, should
Tremble with a start sensitivity etc. all to have a great impact.These factors constrain the polymer gel carrier load energy to insulin further
Power, causes drug loading the least.For drug delivery implant system, low drug loading will significantly reduce gel administration in vivo
Time, cause implanting frequency and rise.This will sharply increase the use cost of such drug-supplying system, reduce the compliance of patient.Also
That is, the mode of loading of existing polymer gel system biopharmaceutical macromolecular drug this kind of to insulin and Drug loading capacity, it is difficult to
Meet the insulin implanted gel carrier performance requirement at aspects such as the most a large amount of load, accurate long-acting controlled releases.
Surface or inside have the porous polymer microsphere of cellular structures and have the biggest specific surface area and hole
Volume, possesses the strongest absorbability inside the surface of microsphere and duct, in terms of the load and controlled release of biopharmaceutical macromolecular drug
There is many unique advantages [Cai Y., Chen Y., Hong X., Liu Z., Yuan W.Porous microsphere and
its applications,Inter.J.Nanomedicine 2013,8,1111-1120.].The loose structure of porous microsphere is used
Oil/water interface, high-speed stirred or the protein denaturation caused such as ultrasonic and rendezvous problem can be avoided in load protein drug.
And, by regulation microsphere size and form, pore passage structure, control load capacity and the distribution of medicine the most relatively easily, but also
Controllable drug releasing rate.These characteristics have made porous polymer microsphere become the ideal carrier of protein drug, but
Being due to the administration controlled fashion of insulin uniqueness, single porous microsphere obviously cannot be administered as the long-acting slow-release of insulin
Carrier.
Described on end, the phenylboric acid functionalized polymer with glucose-sensitive, exploitation is utilized to have similar human pancreas
The prospect that the insulin physiology self-adjustable implantable gel delivery system of function is significant and wide.However it is necessary that solution
Phenylboric acid functionalization hydrogel cannot stimulate and to insulin high-efficient carrier control by response glucose under the physiological condition of pH 7.4
The problem released.
Summary of the invention
The key issue faced as Carriers for Protein Drugs for phenylboric acid functionalized polymer hydrogel, based on system
The thought of engineering, the present invention develops a kind of porous microsphere/polymer of glucose-sensitive under the physiological condition of pH 7.4 of design
Pluralgel system, this pluralgel system is by the implantable positioning performance of gel carrier, stimuli responsive function and porous microsphere
High-efficient carrier characteristic synergism action, be expected to solve insulin physiology self-adjustable implantable gel delivery systems face complexity ask
Topic.
Technical scheme is as follows:
The porous microsphere of a kind of glucose-sensitive/polymer pluralgel, porous microsphere is poly lactic-co-glycolic acid copolymerization
Thing (PLGA) porous microsphere, the average particle size range of PLGA porous microsphere is 1～20 μm；Polymer is containing catechol group
Hyaluronic acid, its architectural feature is as shown in (1) formula:
Described porous microsphere is the PLGA porous microsphere that phenylboric acid function of surface is modified, utilizes aminobenzene boric acid, to PLGA
The modifying surface of porous microsphere, introduces phenylboric acid group.
Described aminobenzene boric acid includes adjacent aminobenzene boric acid, Resocinol-phenol formaldehyde resin and p-aminophenyl boric acid.
The described hyaluronic acid containing catechol group is, utilizes dopamine to hyaluronic acid modified, introduces adjacent benzene two
Phenolic group group, prepare dopamine modified hyaluronic acid, the dopamine grafting efficiency on dopamine modified hyaluronic acid segment be 1～
50%.
Porous microsphere of the present invention/polyhydroxylated polymer pluralgel is: by the porous microsphere of phenylboric acid surface modification with many
The aqueous solution of hydroxy polymer is blended；Between phenylboric acid on microsphere surface and the cis-form dihydroxy on polyhydroxylated polymer segment
There is bonding reaction, form crosslinking polymer network, prepare pluralgel.
The preparation method of the pluralgel of the present invention, comprises the following steps:
(1), phenylboric acid surface modification PLGA porous microsphere is dispersed in water, makes the matter of PLGA porous microsphere aqueous dispersions
Amount concentration range is 1～100%；
(2), by dopamine modified hyaluronic acid it is dissolved in water or phosphatic buffer solution, makes dopamine modified thoroughly
The mass concentration scope of bright matter acid solution is 1～20%；
(3) taking PLGA porous microsphere aqueous dispersions and dopamine modified hyaluronic acid solution respectively, regulation pH value of solution is
6.5-11.5, control temperature, at 4-60 DEG C, stirs, and room temperature stands, and obtains PLGA porous microsphere/dopamine modification hyalomitome
Acid pluralgel；In obtained PLGA porous microsphere/dopamine modified hyaluronic acid pluralgel, dopamine modified hyaluronic acid
Mass content scope is 5～20%, and the mass range of PLGA microspheres amount is 1～20%.
The porous microsphere of the glucose-sensitive of the present invention/polymer pluralgel is applied to drug delivery system；Porous is micro-
Compound the coagulating of sphere/polymer can be implanted, under the physiological condition of pH 7.4 by injection or Minimally Invasive Surgery, it is possible to response
The change of concentration of glucose in environment, it is achieved the controllable release of insulin medicament.
The porous microsphere of glucose-sensitive/polymer pluralgel is applied to develop insulin implanted gel long-acting slow-release system
Agent.
It is described as follows:
PLGA porous microsphere of the present invention is based on emulsion-solvent evaporation method, the porous of preparation in the presence of porogen
Microsphere.Concretely comprising the following steps and be dissolved in suitable quantity of water by porogen, PLGA is dissolved in organic solvent, after the two mixing, utilizes cell to break
Broken instrument emulsifying forms W1 (interior aqueous phase)/O (oil phase) emulsion, and emulsion is scattered in polyvinyl alcohol (PVA) aqueous solution subsequently
(W2) in, forming W1/O/W2 double breast system, solvent flashing under room temperature, washing removes porogen, obtains PLGA porous microsphere.Many
The average particle size range of hole microsphere is 1～20 μm.
PLGA molecular weight ranges of the present invention be the ratio of 5000-50000g/mol, lactic acid and hydroxyacetic acid be 50:50
～85:15.
Porogen used by the PLGA of preparation porous microsphere of the present invention includes ammonium hydrogen carbonate, bovine serum albumin (BSA), oil
Acid sodium.
Organic solvent used by the PLGA of preparation porous microsphere of the present invention includes dichloromethane, chloroform etc..
The PLGA porous microsphere of phenylboric acid surface modification of the present invention be utilize PLGA porous microsphere surface carboxyl and
Reaction between the amino of aminobenzene boric acid, under the effect of a certain amount of catalyst, prepares in the reaction of middle water.
The PLGA porous microsphere of phenylboric acid surface modification of the present invention is to utilize ethylenediamine by PLGA porous microsphere surface
Carboxyl carry out amination after, then at PLGA microsphere surface grafted polyacrylic acid, recycling polyacrylic acid segment on carboxyl
Reacting with the amino of aminobenzene boric acid, under the effect of a certain amount of catalyst, aminobenzene boric acid is keyed to PLGA by Yu Shuizhong
On microsphere surface, prepare the PLGA porous microsphere of phenylboric acid surface modification.
Aminobenzene boric acid of the present invention includes p-aminophenyl boric acid and Resocinol-phenol formaldehyde resin and adjacent aminobenzene boric acid.
Polyacrylic acid molecular weight of the present invention is 1000～5000g/mol.
Polymer containing catechol group of the present invention is the hyaluronic acid that dopamine is modified.
Dopamine modified hyaluronic acid of the present invention is to utilize between the amino of dopamine and the carboxyl of hyaluronic acid
Reaction, sub-at a certain amount of 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (EDC) and N-maloyl
In the presence of amine (NHS), dopamine is keyed on hyaluronic acid by Yu Shuizhong, prepares dopamine modified hyaluronic acid, and its structure is special
Levy as shown in Equation 1.
The process of PLGA porous microsphere load insulin of the present invention is that prepared porous microsphere is scattered in pancreas
In the saturated aqueous solution of island element, fully adsorbing, centrifugation, lyophilization prepares.
The preparation of the porous microsphere of glucose-sensitive of the present invention/polymer pluralgel is by appropriate phenylboric acid table
Modification PLGA porous microsphere in face is dispersed in water, subsequently with a certain amount of dopamine modified hyaluronic acid aqueous solution, and stirring
Uniformly, regulate pH value 6.5-11.5, make dopamine modified hyaluronic acid cross-link, prepare PLGA porous microsphere/polymer compound solidifying
Glue.
The dopamine modified hyaluronic acid water prepared used by PLGA porous microsphere/polymer pluralgel of the present invention
The concentration range of solution is 1～20% (weight).
The PLGA porous microsphere aqueous dispersions prepared used by PLGA porous microsphere/polymer pluralgel of the present invention
Concentration range be 1～100% (weight).
In preparation PLGA porous microsphere/polymer pluralgel of the present invention, containing of dopamine modified hyaluronic acid
Measure in the range of 5～20% (weight).
In preparation PLGA porous microsphere/polymer pluralgel of the present invention, PLGA microspheres amount in the range of 1～
20% (weight).
The porous microsphere of glucose-sensitive of the present invention/polymer pluralgel can be in physiological conditions
(pH7.4) response glucose concentration change occurs swelling-contraction to change, it is achieved continue the long-term of physiology self-adjustable of insulin
Release.
The porous microsphere of glucose-sensitive of the present invention/polymer pluralgel system can be by injecting or micro-
Invasive procedures implants.
The porous microsphere of glucose-sensitive of the present invention/polymer pluralgel system can degradation in vivo.
Porous microsphere of the present invention/polymer pluralgel has glucose-sensitive performance, it is possible in response environment
The change of concentration of glucose, it is achieved the controllable release of insulin, as shown in Figure 1.Phenylboric acid on microsphere surface and dopamine
There is bonding reaction between catechol group on modified hyaluronic acid segment, form cross linked polymer pluralgel.Meanwhile,
Hyaluronic acid be crosslinked be anchored on porous microsphere surface formed closely, the Intelligent controlled release layer of glucose-sensitive.Deposit without glucose
Under, this Intelligent controlled release layer is highly stable, it is possible to stop insulin to discharge from porous microsphere, but the feelings occurred at glucose
Under condition, owing to small molecule glucose can compete the phenylboric acid group of microsphere surface, hyaluronic acid is caused to be peeled off from microsphere surface,
On the one hand destroy the cross-linked network of gel, make gel expand；On the other hand destroy the controlled release layer of microsphere surface, open
Drug release passage, thus cause insulin releasing.The release of insulin, makes internal concentration of glucose reduce, microsphere surface
On phenylboric acid and the catechol group on dopamine modified hyaluronic acid segment between again there is bonding reaction, formed multiple
Close gel, and re-form controlled release layer at microsphere surface, control the release of insulin.
The porous microsphere of glucose-sensitive provided by the present invention/polymer pluralgel system can be planted at exploitation insulin
Enter long-acting slow-release preparation, before providing the aspect such as non-injection formula administering mode of safety convenient to have wide application for diabetics
Scape.
It is a feature of the present invention that: in (1) pluralgel of the present invention, the PLGA porous of phenylboric acid surface modification is micro-
Ball not only plays crosslinking polyhydroxylated polymer and forms the effect of gel, is additionally, since porous polymer microsphere and has the biggest ratio
Surface area and pore volume, it is possible to a large amount of load insulins, effectively solve the macromolecular drugs such as insulin efficiently bearing in gel
Load problem；(2), in pluralgel of the present invention, dopamine modified hyaluronic acid the most still forms the matrix of gel network,
The Intelligent controlled release layer of glucose-sensitive can also be formed, it is achieved the insulin realizing controlled-release of glucose-sensitive on porous microsphere surface
Put；(3) can be between PLGA porous microsphere and the dopamine modified hyaluronic acid of phenylboric acid surface modification of the present invention
Realizing crosslinking in pH7.4 environment, solving polyalcohol hydrogel based on phenylboric acid cannot cross-link in physiological conditions and respond Portugal
The problem that grape sugar stimulates；(4) pluralgel of the present invention has given full play to porous microsphere, glucose-sensitive phenylboric acid, height
The respective function of molecular gel and cooperative effect, be expected to realize the long-term sustained release of insulin physiology self-adjustable, for diabetes
Patient develops efficiently, administration new technique and novel form provide new departure easily.Development field at treating diabetes novel form
There is bigger using value.
Accompanying drawing explanation
Fig. 1 is PLGA porous microsphere/polymer pluralgel and response glucose concentration regulation and control insulin releasing principle thereof
Schematic diagram
Fig. 2 is typical PLGA porous microsphere scanning electron microscope (SEM) photograph
Fig. 3 is the preparation process that the most modified method prepares phenylboric acid surface modification PLGA porous microsphere
Fig. 4 is the PLGA porous microsphere (PLGA-NH of PLGA porous microsphere (PLGA), surface amination2) and surface grafting
The Zeta-potential of polyacrylic PLGA porous microsphere (PLGA-PAA) characterizes
Fig. 5 is the PLGA porous microsphere (PLGA-NH of PLGA porous microsphere (PLGA), face grafted polyacrylic acid2) and benzene boron
The Infrared Characterization of acid surfaces modification PLGA porous microsphere (PLGA-PAAPBA)
Fig. 6 is hyaluronic acid and dopamine modified hyaluronic acid nuclear-magnetism sign
Fig. 7 is that dopamine modified hyaluronic acid aqueous solution and PLGA porous microsphere/dopamine modified hyaluronic acid are compound solidifying
Glue figure
Fig. 8 is the insulin releasing curve of PLGA porous microsphere/dopamine modified hyaluronic acid pluralgel, pH7.4
Detailed description of the invention
Contribute to understanding the present invention by following embodiment, but be not limiting as the summary of the invention of this patent.
Embodiment 1: the preparation of the PLGA porous microsphere of different-shape
(1) PLGA porous microsphere is prepared with BSA for porogen
The PLGA polymer 500mg that the ratio weighing lactic acid and hydroxyacetic acid is 50:50, molecular weight is 5000g/mol, is dissolved in
In 3mL dichloromethane.Weigh 400mg porogen BSA subsequently to be dissolved in 0.6mL water, by dichloromethane solution and the BSA water of PLGA
Solution cell crushing instrument emulsifying 12 times, forms W1 (interior aqueous phase)/O (oil phase) emulsion.After allowing, this emulsion is distributed to
In 300mL 0.4%PVA aqueous solution, high speed shear (7000r/min), form W1/O/W2 double breast system, volatilize under room temperature organic
Solvent, centrifugal drying, obtain PLGA porous microsphere, the mean diameter of microsphere is about 20 μm.
(2) PLGA porous microsphere is prepared with ammonium hydrogen carbonate for porogen
The PLGA polymer 500mg that the ratio weighing lactic acid and hydroxyacetic acid is 75:25, molecular weight is 30000g/mol, molten
In 3mL chloroform.Weigh 10mg porogen ammonium hydrogen carbonate subsequently to be dissolved in 1mL water, by the chloroform soln of PLGA and
Ammonium bicarbonate aqueous solution cell crushing instrument emulsifying 20 times, forms W1 (interior aqueous phase)/O (oil phase) emulsion.By this milkiness after allowing
Liquid is distributed in 300mL 0.4%PVA aqueous solution, high speed shear (10000r/min), forms W1/O/W2 double breast system, room temperature
Lower volatile organic solvent, centrifugal drying, obtain PLGA porous microsphere, the mean diameter of microsphere is about 10 μm.
(3) PLGA porous microsphere is prepared with enuatrol for porogen
The PLGA polymer 500mg that the ratio weighing lactic acid and hydroxyacetic acid is 85:15, molecular weight is 50000g/mol, molten
In 3mL dichloromethane.Weigh 10mg porogen enuatrol subsequently to be dissolved in 1mL water, by dichloromethane solution and the oil of PLGA
Acid sodium aqueous solution cell crushing instrument emulsifying 20 times, forms W1 (interior aqueous phase)/O (oil phase) emulsion.After allowing, this emulsion is divided
It is scattered in 300mL 0.4%PVA aqueous solution, high speed shear (10000r/min), forms W1/O/W2 double breast system, wave under room temperature
Sending out organic solvent, centrifugal drying, obtain PLGA porous microsphere, the mean diameter of microsphere is about 5 μm.Use scanning electron microscopic observation PLGA
The pattern of porous microsphere, is coated in the PLGA porous microsphere after lyophilizing on conducting resinl, takes the section of a small amount of microsphere simultaneously, uses FEI
Nano 450 observes surface topography and the section thereof of PLGA porous microsphere.As shown in Figure 2, in figure, result shows test result,
The mean diameter of PLGA porous microsphere, at 5 μm, rough surface, has aperture.Microsphere section result shows, in irregularly inside PLGA
Loose structure.
(4) PLGA porous microsphere is prepared with enuatrol for porogen
The PLGA polymer 500mg that the ratio weighing lactic acid and hydroxyacetic acid is 50:50, molecular weight is 20000g/mol, molten
In 3mL dichloromethane.Weigh 5mg porogen enuatrol subsequently to be dissolved in 0.5mL water, by dichloromethane solution and the oil of PLGA
Acid sodium aqueous solution cell crushing instrument emulsifying 20 times, forms W1 (interior aqueous phase)/O (oil phase) emulsion.Form W1 (interior aqueous phase)/O
(oil phase) emulsion.This emulsion is distributed in 300mL 0.4%PVA aqueous solution after allowing, high speed shear (10000r/min),
Form W1/O/W2 double breast system, volatile organic solvent under room temperature, centrifugal drying, obtain PLGA porous microsphere, the average particle of microsphere
Footpath is about 1 μm.
Embodiment 2: the preparation of phenylboric acid surface modification PLGA porous microsphere
(1) the most modified method prepares phenylboric acid surface modification PLGA porous microsphere
The most modified method prepares the process of phenylboric acid surface modification PLGA porous microsphere as shown in Figure 3, first with second two
The carboxyl on PLGA porous microsphere surface is carried out amination by amine, and then at PLGA microsphere surface grafted polyacrylic acid, recycling is poly-
Carboxyl on acrylate reacts with the amino of aminobenzene boric acid, is keyed on PLGA microsphere surface by aminobenzene boric acid, system
Obtain the PLGA porous microsphere of phenylboric acid surface modification.Aminobenzene boric acid includes p-aminophenyl boric acid, Resocinol-phenol formaldehyde resin and adjacent ammonia
Base phenylboric acid, wherein optimum selection is Resocinol-phenol formaldehyde resin.Detailed process is as follows, is scattered in by the PLGA porous microsphere of 250mg
In MES aqueous solution (MES, 0.1mol/L, pH=5.5), it is subsequently added a certain amount of catalyst 1-(3-dimethylamino
Propyl group)-3-ethyl-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS), activation 2h is stirred at room temperature, adds
500mg ethylenediamine, reacts 24h, centrifugation, washs 3-5 time, prepares the PLGA microsphere of surface amination.By amidized
PLGA microsphere is scattered in appropriate polyacrylic acid aqueous solution, and acrylic acid optimum weight scope is 1000～5000g/mol.With
After, adding appropriate EDC and NHS, centrifugation after 4 DEG C of reaction 24h, the PLGA that washing prepares surface-grafted polyacrylic acid is micro-
Ball.Subsequently the PLGA microsphere 500mg obtaining surface-grafted polyacrylic acid is scattered in appropriate aqueous solution, add EDC and
NHS, after stir-activating, adds the Resocinol-phenol formaldehyde resin of 250mg, reacts 24h, centrifugation, and washing, vacuum drying is the most available
Phenylboric acid surface modification PLGA porous microsphere.
To obtained PLGA porous microsphere (PLGA), the PLGA porous microsphere (PLGA-NH of surface amination2) and surface
The Zeta-potential of the PLGA porous microsphere (PLGA-PAA) of grafted polyacrylic acid characterizes, as shown in Figure 4.Test result shows
Show, PLGA, PLGA-NH2, the Zeta-potential of PLGA-PAA porous microsphere is respectively-22.6 ± 1.8mV ,-8.7 ± 0.67mV ,-
30.1±2mV.The change of PLGA-PAA porous microsphere surface charge shows that the carboxyl-content on microsphere surface increases, PLGA-PAA
Successfully prepared.PLGA porous microsphere (PLGA), the PLGA porous microsphere (PLGA-PAA) of surface-grafted polyacrylic acid and benzene boron
The Infrared Characterization of the PLGA porous microsphere (PLGA-PAAPBA) that acid surfaces is modified, as shown in Figure 5.Test result shows, relatively
In PLGA porous microsphere, the carboxyl-content of PLGA-PAA porous ball becomes many, and PLGA-PAAPBA is at 1615cm-1, 1530cm-1Occur
On phenyl ring, the stretching vibration peak of C=C, illustrates that microsphere surface has connected phenylboric acid group.
(2) the most modified method prepares phenylboric acid surface modification PLGA porous microsphere
The PLGA porous microsphere of 500mg is scattered in MES buffer (0.1mol/L pH=5.5), is subsequently added certain
EDC and NHS of amount, is stirred at room temperature activation 4h, is subsequently added the aminobenzene boric acid of 500mg, react under conditions of ice-water bath
24h, centrifugation, with distilled water wash 3-5 time, lyophilization, obtain phenylboric acid surface modification PLGA porous microsphere.
Embodiment 3: the phenylboric acid surface modification PLGA porous microsphere load to insulin
Insulin is scattered in appropriate water, is 1.0-7.4 by the pH value range of dropping dilute hydrochloric acid regulation and control solution, adjusts
Control insulin dissolubility in water, the centrifuging and taking supernatant can obtain the insulin saturated aqueous solution of variable concentrations.Take subsequently
The saturated aqueous solution of 2mL insulin, adds 100mg phenylboric acid surface modification PLGA porous microsphere, gentle agitation 2-24h, is centrifuged
Separating, lyophilization obtains being loaded with the phenylboric acid surface modification PLGA porous microsphere of insulin.By test insulin solutions quilt
The change of concentration before and after micro-ball load, can calculate the PLGA porous microsphere load capacity to insulin.PLGA porous microsphere is to pancreas
The load capacity of island element is about 10～45%.
Embodiment 4: the preparation of dopamine modified hyaluronic acid
Weigh hyaluronic acid (1.5g) to be dissolved in 100mL MES buffer, add EDC (3.19g), NHS (1.92g), DOPA
Amine (2g), 4 DEG C of reactions, seal after logical nitrogen 4h, react 4-48h, dialysis, lyophilizing obtain dopamine modified hyaluronic acid, DOPA
Amine groups grafting efficiency on hyaluronic acid segment is 1～50%.Hyaluronic acid and dopamine modified hyaluronic acid are carried out
Nuclear-magnetism sign, as shown in Figure 6, relative to hyaluronic acid, dopamine modified hyaluronic acid goes out to have result at 6.5-7.5ppm
Significantly benzene ring hydrogen absworption peak, occurs in that, at 3.3ppm, the-CH connected mutually with amino simultaneously2-the absworption peak of hydrogen, explanation
Dopamine modified hyaluronic acid is successfully prepared.
Embodiment 5: the preparation of porous microsphere/polymer pluralgel
Phenylboric acid surface modification PLGA porous microsphere (100mg) is scattered in 1mL water, and the PLGA forming 10% (weight) is micro-
Ball dispersion liquid.Dopamine modified hyaluronic acid (100mg) is dissolved in 2mL phosphate buffered solution or aqueous solution, is formed
The dopamine modified hyaluronic acid solution of 5% (weight).PLGA microsphere dispersion liquid and dopamine modified hyaluronic acid solution are mixed
Close, regulation pH value of solution is 6.5-11.5, control temperature at 4-60 DEG C, stir, room temperature standing, obtain PLGA porous microsphere/
Dopamine modified hyaluronic acid pluralgel.
Prepare the concentration range of the PLGA porous microsphere aqueous dispersions used by porous microsphere/polymer pluralgel be 1～
100% (weight), the optimum selection of the concentration of PLGA porous microsphere aqueous dispersions is 10～20% (weight).Preparation PLGA porous
The concentration range of the dopamine modified hyaluronic acid aqueous solution used by micro-sphere/polymer pluralgel is 1～20% (weight), many
The optimum selection of the concentration of bar amine modified hyaluronic acid aqueous solution is 5～15% (weight).Prepare porous microsphere/polymer to be combined
The pH value range of gel is 6.5-11.5, and wherein pH value optimum selection is neutrallty condition pH=7.4.Prepare porous microsphere/polymerization
The temperature of thing pluralgel is at 4-60 DEG C, and wherein temperature value optimum selection is 15～30 DEG C.
In preparation process, the amount added by regulation PLGA porous microsphere and dopamine modified hyaluronic acid, thus it is possible to vary
The crosslink density of pluralgel, mechanical property and drug loading.Last obtained PLGA porous microsphere/dopamine modified hyaluronic acid
In pluralgel, the OK range of dopamine modified hyaluronic acid content is 5～20% (weight), and the model of PLGA microspheres amount
Enclose is 1～20% (weight).
Dopamine modified hyaluronic acid aqueous solution and PLGA porous microsphere/dopamine modified hyaluronic acid pluralgel are the most attached
Shown in Fig. 7, relative to dopamine modified hyaluronic acid aqueous solution, PLGA porous microsphere/dopamine modified hyaluronic acid is compound solidifying
Glue is white, and not the flowing of gel after being inverted by vial, illustrates that dopamine modified hyaluronic acid is crosslinked, defines
Pluralgel.
The insulin releasing of embodiment 6:PLGA porous microsphere/dopamine modified hyaluronic acid pluralgel
By modified for the dopamine of the aqueous dispersions 1mL and 10% of the PLGA porous microsphere being loaded with insulin that concentration is 10%
Phosphate buffered solution 3mL of hyaluronic acid is blended, and pH is 7.4, stirs, and the PLGA porous obtaining being loaded with insulin is micro-
Ball/dopamine modified hyaluronic acid pluralgel.
Take the above-mentioned gel of 0.5mL respectively to be placed in bag filter (molecular cut off: 8000), subsequently bag filter is immersed 50mL
Concentration of glucose is respectively 0mg/mL, and in the phosphate buffered solution of 5mg/mL, 10mg/mL, pH value is 7.4, by shaking table temperature
Being set to 37 DEG C, respectively at 15min, 30min, 45min, 60min, 90min, 120min, 180min sample, and use uv-spectrophotometric
Meter is observed and is surveyed absorbance at 226.5.Under different concentration of glucose, insulin from pluralgel vitro cumulative burst size-time
Half interval contour is as shown in Figure 8, it can be seen that without in D/W, glucose does not discharges, 24 hours Cumulative release amounts
It is about 5%；Along with the addition of glucose, insulin releasing is substantially accelerated, and with the rising of concentration of glucose, releasing of insulin
High-volume increase.This gel rubber system achieves in physiological conditions, with the change of glucose content, the release of regulation and control insulin.
Claims (7)
1. the porous microsphere of a glucose-sensitive/polymer pluralgel, it is characterised in that porous microsphere is polylactic acid-glycolic base
Acetate multipolymer (PLGA) porous microsphere, the average particle size range of PLGA porous microsphere is 1～20 μm；Polymer is for containing adjacent benzene
The hyaluronic acid of diphenol group, shown in its following structural features:
2. the pluralgel described in claim 1, it is characterised in that porous microsphere is that the PLGA of phenylboric acid function of surface modification is many
Hole microsphere, utilizes aminobenzene boric acid, the modifying surface to PLGA porous microsphere, introduces phenylboric acid group.
3. the pluralgel described in claim 1, it is characterised in that aminobenzene boric acid includes adjacent aminobenzene boric acid, m-aminophenyl boron
Acid and p-aminophenyl boric acid.
4. the pluralgel described in claim 1, it is characterised in that the described hyaluronic acid containing catechol group is, utilizes
Dopamine, to hyaluronic acid modified, introduces catechol group, prepares dopamine modified hyaluronic acid, dopamine modification hyalomitome
Dopamine grafting efficiency on acid segment is 1～50%.
5. the preparation method of the pluralgel of claim 1, it is characterised in that comprise the following steps:
(1), being dispersed in water by phenylboric acid surface modification PLGA porous microsphere, the quality making PLGA porous microsphere aqueous dispersions is dense
Degree scope is 1～100%；
(2), by dopamine modified hyaluronic acid it is dissolved in water or phosphatic buffer solution, makes dopamine modification hyalomitome
The mass concentration scope of acid solution is 1～20%；
(3) taking PLGA porous microsphere aqueous dispersions and dopamine modified hyaluronic acid solution respectively, regulation pH value of solution is 6.5-
11.5, control temperature, at 4-60 DEG C, stirs, and room temperature stands, and obtains PLGA porous microsphere/dopamine modified hyaluronic acid multiple
Close gel；In obtained PLGA porous microsphere/dopamine modified hyaluronic acid pluralgel, dopamine modified hyaluronic acid quality
Content range is 5～20%, and the mass range of PLGA microspheres amount is 1～20%.
6. the porous microsphere of glucose-sensitive/polymer pluralgel is applied to drug delivery system；Porous microsphere/polymer is multiple
Close and coagulate and can be implanted, under the physiological condition of pH 7.4 by injection or Minimally Invasive Surgery, it is possible to Fructus Vitis viniferae in response environment
The change of sugar concentration, it is achieved the controllable release of insulin medicament.
7. the porous microsphere of glucose-sensitive/polymer pluralgel is applied to develop insulin implanted gel long-acting slow-release system
Agent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610493506.XA CN106038478B (en) | 2016-06-24 | 2016-06-24 | A kind of porous microsphere of glucose-sensitive/polymer plural gel and its preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610493506.XA CN106038478B (en) | 2016-06-24 | 2016-06-24 | A kind of porous microsphere of glucose-sensitive/polymer plural gel and its preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106038478A true CN106038478A (en) | 2016-10-26 |
| CN106038478B CN106038478B (en) | 2019-06-04 |
Family
ID=57166839
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610493506.XA Expired - Fee Related CN106038478B (en) | 2016-06-24 | 2016-06-24 | A kind of porous microsphere of glucose-sensitive/polymer plural gel and its preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106038478B (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106729638A (en) * | 2016-12-14 | 2017-05-31 | 西安石油大学 | A kind of preparation method of poly-dopamine microcapsules oral insulin drug administration carrier |
| CN108079282A (en) * | 2018-01-25 | 2018-05-29 | 武汉大学 | A kind of intelligence uelralante adjusts gold nanoclusters particle of blood glucose and preparation method thereof |
| CN108175765A (en) * | 2017-12-01 | 2018-06-19 | 东南大学 | A kind of acid-sensitive controlled release anti-inflammatory gel and preparation method and application |
| CN108635570A (en) * | 2018-06-06 | 2018-10-12 | 武汉大学 | A kind of gold nanoclusters particle of response type blood glucose-control and preparation method thereof |
| CN110194877A (en) * | 2019-03-20 | 2019-09-03 | 湘潭大学 | A kind of Nanometer composite hydrogel and its preparation method and application |
| CN111658783A (en) * | 2020-07-09 | 2020-09-15 | 中国药科大学 | Switch type glucose responsive double-layer cross-linked polymer micelle drug delivery system and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001054593A1 (en) * | 2000-01-31 | 2001-08-02 | The General Hospital Corporation | Neural regeneration conduit |
| CN102596275A (en) * | 2009-09-04 | 2012-07-18 | 亚洲大学校产学协力团 | In situ-forming hydrogel for tissue adhesives and biomedical use thereof |
| CN104840415A (en) * | 2014-02-19 | 2015-08-19 | 香港浸会大学 | Long-acting controlled release liposome gel combination containing blood sugar reducing active component, and preparation method thereof |
-
2016
- 2016-06-24 CN CN201610493506.XA patent/CN106038478B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001054593A1 (en) * | 2000-01-31 | 2001-08-02 | The General Hospital Corporation | Neural regeneration conduit |
| CN102596275A (en) * | 2009-09-04 | 2012-07-18 | 亚洲大学校产学协力团 | In situ-forming hydrogel for tissue adhesives and biomedical use thereof |
| CN104840415A (en) * | 2014-02-19 | 2015-08-19 | 香港浸会大学 | Long-acting controlled release liposome gel combination containing blood sugar reducing active component, and preparation method thereof |
Non-Patent Citations (3)
| Title |
|---|
| FULI ZHAO等: "An injectable particle-hydrogel hybrid system for glucose-regulatory insulin delivery", 《ACTA BIOMATERIALIA》 * |
| HYUN-JI PARK等: "Catechol-Functionalized Hyaluronic Acid Hydrogels Enhance Angiogenesis and Osteogenesis of Human Adipose-Derived Stem Cells in Critical Tissue Defects", 《BIOMACROMOLECULES》 * |
| 左芳等: "基于苯硼酸-邻苯二酚相互作用的聚合物凝胶的研究", 《西南民族大学学报自然科学版》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106729638A (en) * | 2016-12-14 | 2017-05-31 | 西安石油大学 | A kind of preparation method of poly-dopamine microcapsules oral insulin drug administration carrier |
| CN108175765A (en) * | 2017-12-01 | 2018-06-19 | 东南大学 | A kind of acid-sensitive controlled release anti-inflammatory gel and preparation method and application |
| CN108079282A (en) * | 2018-01-25 | 2018-05-29 | 武汉大学 | A kind of intelligence uelralante adjusts gold nanoclusters particle of blood glucose and preparation method thereof |
| CN108079282B (en) * | 2018-01-25 | 2020-06-09 | 武汉大学 | Gold nanocluster particles capable of intelligently releasing insulin to regulate blood sugar and preparation method of gold nanocluster particles |
| CN108635570A (en) * | 2018-06-06 | 2018-10-12 | 武汉大学 | A kind of gold nanoclusters particle of response type blood glucose-control and preparation method thereof |
| CN110194877A (en) * | 2019-03-20 | 2019-09-03 | 湘潭大学 | A kind of Nanometer composite hydrogel and its preparation method and application |
| CN110194877B (en) * | 2019-03-20 | 2021-07-20 | 湘潭大学 | Nano composite hydrogel and preparation method and application thereof |
| CN111658783A (en) * | 2020-07-09 | 2020-09-15 | 中国药科大学 | Switch type glucose responsive double-layer cross-linked polymer micelle drug delivery system and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106038478B (en) | 2019-06-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106038478B (en) | A kind of porous microsphere of glucose-sensitive/polymer plural gel and its preparation method and application | |
| EP1750679B1 (en) | Gel composition comprising charged polymers | |
| JP2001517494A (en) | Improved hydrogels for tissue engineering | |
| CN101148520B (en) | Temperature sensitive chitosan hydrogel | |
| CN107496382A (en) | Composite Nano capsule injection aquagel dual drug-loading slow-releasing system and preparation method | |
| CN114146046A (en) | Coated microneedle with multilayer structure, preparation method thereof and microneedle patch comprising coated microneedle | |
| CN107811963A (en) | Soluble microneedle device and its application | |
| CN102357077B (en) | Protein nanometer particle for wrapping slightly soluble medicines and preparation method thereof | |
| CN111870806A (en) | Magnetic control microneedle robot and preparation method, use method and application thereof | |
| CN106821963A (en) | A kind of method of utilization cellulose base temperature and the load of pH sensitive hydrogels and slow releasing pharmaceutical | |
| CN103126975B (en) | Preparation method of hydrogel patch substrate with gradient drug concentration | |
| CN105596298A (en) | PEG-PLGA sustained release microsphere with encapsulated buprenorphine and preparation method thereof | |
| CN115317665B (en) | Polyester particle composite temperature-sensitive instant gel subcutaneous implant | |
| CN108186602A (en) | Two-layer compound sustained-release micro-spheres prevent and treat osteoporosis in preparation, and arthritis and cartilage restore the application in drug | |
| CN112807439A (en) | Preparation method and application of implantable in-situ formed chitosan hydrogel | |
| CN102357076B (en) | Preparation method of protein nanoparticles coating insoluble drug | |
| CN107233296A (en) | Thymopeptide-5 solubility micropin and preparation method thereof | |
| CN110063946A (en) | A kind of chitosan sodium alginate micro ball preparation method and application for containing Ah pa and replacing Buddhist nun | |
| CN115746341A (en) | Polymer microsphere, preparation method thereof and application thereof in injection type tissue filler | |
| CN114146050B (en) | Soluble microneedle substrate material and preparation method and application thereof | |
| CN115804842A (en) | Dacarbazine nano microneedle preparation as well as preparation method and application thereof | |
| Manna et al. | Interpenetrating polymer network in microparticulate systems: drug delivery and biomedical application | |
| CN108159021A (en) | A kind of two-layer compound sustained-release micro-spheres for being loaded with promoting bone growing albumen and eliminating arthritic symptom drug and preparation method thereof | |
| CN116672498A (en) | Composite material for injection and application thereof | |
| CN113197842A (en) | Cannabidiol injectable hydrogel, preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20190604 Termination date: 20210624 |