CN108421049A - Mesoporous carbon particle of insulin and preparation method thereof, oral preparation and application - Google Patents
Mesoporous carbon particle of insulin and preparation method thereof, oral preparation and application Download PDFInfo
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Abstract
The invention discloses a kind of mesoporous carbon particle of insulin, including mesoporous carbon and insulin, the insulin is embedded in mesoporous carbon, and the amount ratio of the insulin and mesoporous carbon is 36:10‑15.Correspondingly, invention additionally discloses the application of oral preparation and the mesoporous carbon particle of above-mentioned insulin in treating diabetes made of a kind of preparation method of the mesoporous carbon particle of insulin and the mesoporous carbon particle of above-mentioned insulin.It using the present invention, prepares simply, good biocompatibility will not cause cell or internal organs toxic reaction, and excretion is very fast in vivo.Oral preparation obtained solves the problems, such as the first pass effect of unstable and liver of the insulin in digestive tract environment, realizes and takes orally, and ensures the oral efficient treatment of polypeptide.
Description
Technical field
The present invention relates to insulin practical technique field more particularly to a kind of mesoporous carbon particle of insulin and its preparation sides
Method, oral preparation and application.
Background technology
At present for diabetic, insulin remains the head for maintaining blood glucose level by promoting blood glucose to take in
Want drug.Even to this day, insulin is still the first-line drug for treating Type I diabetes, also effective in cure to patients with NIDDM.
Insulin is mainly still administered in a manner of hypodermic and cannot be taken orally, and reason is that it is unstable in digestive tract environment,
The first pass effect etc. of influence including hydrochloric acid in gastric juice and protease, lower intestinal wall permeability and liver.But long-term subcutaneous is noted
Penetrating insulin can cause injection site to infect, and mental pressure is caused to increase, and be reduced so as to cause patient compliance,
And then lead to futile treatment.In addition, voluntarily subcutaneous insulin injections are also possible to operation error to patient, lead to excess injection, draw
Play the adverse reactions such as hypoglycemia.Therefore, the whole world is still dedicated to developing novel non-injection insulin medicament so far, passing to overcome
Various disadvantages caused by subcutaneous insulin injections.
Currently, insulin non-injection administration mode is mainly that spray is sucked through lung, but this administering mode is brought
Many problems, for example it causes the asthma of patient or increases the risk etc. of lung cancer.And transdermal means administration is then because of skin
The problem of skin barrier, there are problems that absorbing, working slowly even inefficacy.Therefore, oral administration is returned, is prepared novel
Oral insulin preparation it is particularly necessary.
The obstacle that the research and development of oral insulin preparation are encountered includes mainly：(1) dissolving of gastric juice and protease disappear
Change；(2) enteron aisle stickiness mucous layer；(3) intestinal epithelial cell.Therefore, the R＆D direction of Macrulin is mainly wrapped at present
It includes：(1) absorption of insulin accelerating agent；(2) protease inhibitors；(3) intestinal mucosa adhesive agent；(4) particle drug-loaded system.Always
It, the research direction of oral insulin preparation can be summarized as：(1) prevent insulin from being destroyed by protease；(2) do not changing pancreas
Enhance absorption of the insulin in digestive system under the premise of the element physiological activity of island.
The concrete mode of oral insulin preparation includes：(1) chemical constitution of insulin is modified, avoids being disappeared
Change road enzyme hydrolysis；(2) absorption of insulin reinforcing agent shifts by paracellular pathway, across cell or changes Gut wall epithelial cells
Characteristic breaks Gut barrie r to promote absorption of insulin；(3) enzyme inhibitor；(4) cell-penetrating peptides, by improving plasma membrane to egg
In vain, the penetrability of polypeptide promotes absorption of insulin；(5) polymer system of mucous membrane tackness, this is most promising research and development side
To this kind of material is by changing its mucosa adhesion performance when the environment such as pH, temperature, enzyme, light, electric field or ionic strength, as shell is poly-
Sugar；(6) oral microparticle delivery system, predominantly 1~1000 μm of microencapsulated delivery system, avoid insulin from dissolving out and increase under one's belt
It is strong its in the absorption of enteron aisle, such as beta-cyclodextrin, polylactic acid；(7) oral administration nanometer grain drug delivery system, particle size range be 1~
100nm, such as polymer micelle.
Although above-mentioned oral insulin preparation R＆D direction, which has, improves insulin stability, enhancing absorption of insulin
The advantages of, but due to its that there are costs is excessively high, potential safety and bioavilability be not up to standard the problems such as, U.S. FDA is extremely
The present does not ratify any Macrulin listing yet.Therefore, the research and development of Macrulin must also be carried out, and should
Toward direction that is safe and non-toxic, improving bioavilability research and development.
According to the definition of International Union of Pure and Applied Chemistry (IUPAC), porous material can be divided into poromerics, mesoporous
Material and large pore material.Aperture is large pore material more than 50nm, and what aperture was situated between 2nm~50nm is mesoporous material, and aperture is small
In 2nm be poromerics.Mesoporous material is since with higher specific surface area, hole holds and adjustable aperture structure, pattern
The features such as, there is stronger suction-operated to large biological molecule, heavy metal ion and synthesis organic pollution etc..
The prior art also has prepares insulin granule using mesoporous material, for example, Publication No. CN 102961340B are public
It opens《Nanoscale insulin and preparation method thereof》：It is using aerosil as the carrier of insulin, the titanium dioxide
The porosity of silica aerogel is 95~99%, aperture is 10~50nm, specific surface area is 200~1000m2/ g, density be 3~
300kg/m3, network consisting colloidal particle diameter be 1~50nm, the diameter of the nanoscale insulin particle 100nm with
Under；The nanoscale insulin particle further includes PEG；
The preparation method of the nanoscale insulin particle includes the following steps：
(1) insulin is dissolved in the hydrochloric acid solution of 0.01mol/L；
(2) aerosil is added into above-mentioned hydrochloric acid solution；
(3) dry after insulin and aerosil absorption completely；
(4) PEG is dissolved in absolute ethyl alcohol；
(5) solid of the dry gained of step (3) is added in the ethanol solution of step (4)；
(6) ethanol solution of step (5) is sent into mulser and is emulsified；
(7) emulsion obtained by step (6) is dry in thermostatic drying chamber；
(8) solid of the dry gained of grinding steps (7), and 200 mesh sieve is crossed to get nanoscale insulin particle；
When aerosil described in the step (2) has hydrophobicity, need first to pass through before hydrochloric acid solution is added
300~1000 DEG C of heat treatments make it have hydrophily.
Documents 1 solve the problems, such as that the bioavilability of current oral insulin is low, and which improve nanoparticles to be inhaled
The case where receipts.However, the carrier that documents 1 use is aerosil, cell or internal organs toxic reaction may be caused,
It is easy to accumulate in vivo.And the nanoscale insulin particle of documents 1 prepares complexity.
Invention content
Technical problem to be solved by the present invention lies in provide a kind of mesoporous carbon particle of insulin and preparation method thereof, system
Standby simple, good biocompatibility will not cause cell or internal organs toxic reaction, and excretion is very fast in vivo.
Technical problem to be solved by the present invention lies in, oral preparation made of a kind of mesoporous carbon particle of insulin is provided,
The oral preparation solves the problems, such as the first pass effect of unstable and liver of the insulin in digestive tract environment, realizes and takes orally,
And ensure the oral efficient treatment of polypeptide.
Technical problem to be solved by the present invention lies in provide a kind of mesoporous carbon particle of insulin in treating diabetes
Using.
In order to solve the above technical problem, the present invention provides a kind of mesoporous carbon particles of insulin, including mesoporous carbon and pancreas
Island element, the insulin are embedded in mesoporous carbon, and the amount ratio of the insulin and mesoporous carbon is 3-6:10-15.
As the improvement of said program, the amount ratio of the insulin and mesoporous carbon is 4-5:11-13.
As the improvement of said program, the grain size of the mesoporous carbon is 2nm~50nm.
As the improvement of said program, the grain size of the mesoporous carbon is 10nm~40nm.
As the improvement of said program, the mesoporous carbon selects MCN-1, MCN-2 or MCN-3, wherein
MCN-1 refers to that specific surface area is 755-765m2/ g, Kong Rongwei 0.8-0.95cm3/ g, pore-size distribution 3.9-7.8
The mesoporous carbon of nm；
MCN-2 refers to that specific surface area is 740-754m2/ g, Kong Rongwei 0.96-1.15cm3/ g, pore-size distribution 3.9-7.6
The mesoporous carbon of nm；
MCN-3 refers to that specific surface area is 480-530m2/ g, Kong Rongwei 0.50-0.80cm3/ g, pore-size distribution 8.0-
The mesoporous carbon of 20nm.
Correspondingly, the present invention discloses a kind of preparation method such as the mesoporous carbon particle of insulin, including:
10-15mg mesoporous carbons and 3-6mg insulin are placed in test tube, the alcoholic solution and 0.1- of 0.4-1.2mL is added
The acid solution of 0.4mL, a concentration of 0.005-0.03mol/L of acid solution, vortex oscillation 20-50min；
Sample after oscillation is transferred in evaporating dish, and room temperature drying obtains the mesoporous carbon particle of insulin.
As the improvement of said program, the dosage of the mesoporous carbon is 11-13mg, and the dosage of the insulin is 4-5mg,
The dosage of the alcoholic solution is 0.6-1.0mL, and the dosage of the acid solution is 0.2-0.3mL, a concentration of 0.01- of acid solution
The time of 0.02mol/L, vortex oscillation are 25-35min.
As the improvement of said program, it is hydrochloric acid or acetic acid that the alcoholic solution, which selects methanol or ethyl alcohol, the acid solution,.
Correspondingly, the present invention also provides the insulin oral preparations made of above-mentioned insulin mesoporous carbon particle.
Correspondingly, the application the present invention also provides the mesoporous carbon particle of insulin in treating diabetes.
Implement the present invention, has the advantages that：
One, a kind of mesoporous carbon particle of insulin of present invention offer, including mesoporous carbon and insulin, insulin and mesoporous carbon
Amount ratio is 3-6:10-15, moreover, the present invention passes through specific preparation method so that insulin molecule is integrated into mesoporous carbon
Duct, and then realize and carry medicine, success insulin synthesis mesoporous carbon nanoparticle.
Two, the preparation method of the mesoporous carbon particle of the insulin is simple, only includes two steps, and alcoholic solution and acid are molten
Liquid is added simultaneously inside a step, simplifies operating procedure, saves preparation time.
Three, adsorption rate of the invention is respectively 25-55%, and drugloading rate is respectively 8-15%, meets pharmacodynamics demand；Release
Degree is 15-57%, realizes being released effectively in simulated intestinal fluid environment；Blood glucose in diabetic rats significantly drops after oral medication 4h
It is low, have the effect of sustained release compared with being subcutaneously injected；Has certain adsorption capacity to colon cancer cell, insulin mesoporous carbon is received
The grain of rice can be realized in intestinal cell and be transported.
Four, complicated with preparation, biocompatibility is poor, may cause cell or internal organs toxic reaction and be easy to store in vivo
Long-pending mesoporous silicon oxide is compared, and meso-porous carbon material toxicity of the invention is relatively low, is had good biocompatibility, is drained in vivo
Comparatively fast, preparation method is easy, and yield is higher.
Description of the drawings
Fig. 1 is the SEM spectrum (A.MCN-1 of blank mesoporous carbon sample and insulin mesoporous carbon sample；B. MCNI-1；
C.MCN-2；D.MCNI-2；E.MCN-3；F.MCNI-3)；
Fig. 2 is the TEM collection of illustrative plates (A.MCN-1 of blank mesoporous carbon sample and insulin mesoporous carbon sample；B. MCNI-1；
C.MCN-2；D.MCNI-2；E.MCN-3；F.MCNI-3)；
Fig. 3 A are the FTIR spectrum (A. of insulin, blank mesoporous carbon sample and insulin mesoporous carbon nanoparticle
Insulin；B.MCN-1；C.MCNI-1；D.MCN-2)；
Fig. 3 B are the FTIR spectrum (E. of insulin, blank mesoporous carbon sample and insulin mesoporous carbon nanoparticle
MCNI-2；F.MCN-3；G.MCNI-3)；
Fig. 4 is the schematic diagram of release of the insulin mesoporous carbon nanoparticle in pH7.4PBS solution；
Fig. 5 be Oral Administration in Rats insulin mesoporous carbon nanoparticle after blood glucose percent change value schematic diagram；
Fig. 6 is that the meso-porous carbon material of FITC fluorescent markers acts on the schematic diagram of colon cancer HCT-116 cells.
Specific implementation mode
To make the object, technical solutions and advantages of the present invention clearer, the present invention is made into one below in conjunction with attached drawing
Step ground detailed description.
The present invention provides a kind of mesoporous carbon particle of insulin, including mesoporous carbon and insulin, the insulin is embedded in
In mesoporous carbon, the amount ratio of the insulin and mesoporous carbon is 3-6:10-15.Preferably, the use of the insulin and mesoporous carbon
Amount is than being 4-5:11-13.More preferably, the insulin and the amount ratio of mesoporous carbon are 4:12.
Mesoporous carbon is a kind of novel non-silicon-based mesoporous material, 2nm<Aperture<50nm has huge specific surface area (can
Up to 2500m2/ g) and pore volume (up to 2.25cm3/ g), to provide the drugloading rate of bigger.Compared with pure mesoporous silicon material,
Meso-porous carbon material shows special property, there is high specific surface area, high porosity；Aperture size is adjustable in a certain range；
Mesoporous various shapes, hole wall composition, structure and property are adjustable；High thermal stability and hydro-thermal can be obtained by optimum synthesis condition
Stability；Synthesize simple, easy to operate, physiological-toxicity-free.Wherein, the grain size of the mesoporous carbon is 2nm~50nm.Preferably, institute
The grain size for stating mesoporous carbon is 10nm~40nm.More preferably, the grain size of the mesoporous carbon is 10nm~40nm.
The insulin and the amount ratio of mesoporous carbon are 3-6:The mass ratio of 10-15, suitable insulin and mesoporous carbon is
Ensure the guarantee of its release performance and carrier medicine carrying efficiency.If the content of insulin is relatively low, it is likely that lead to the release speed of insulin
Rate slows down；If insulin content is higher, it is likely that cause insulin releasing to accelerate, and cause to carry medicine in vain, to proinsulin
Expect the waste of medicine.
The insulin and mesoporous carbon of the present invention passes through specific preparation method with specific dosage so that insulin point
Son is integrated into the duct of mesoporous carbon, and then realizes and carry medicine, success insulin synthesis mesoporous carbon nanoparticle.Insulin after synthesis is situated between
Hole carbon nanoparticle can overcome the obstructions such as the dissolving of gastric juice and the digestion of protease, enteron aisle stickiness mucous layer, intestinal epithelial cell, suitable
Profit is discharged in enteron aisle, and blood glucose in diabetic rats significantly reduces after 4h is administered orally, and has the effect of sustained release compared with being subcutaneously injected.
Specifically, the mesoporous carbon can select MCN-1, MCN-2 or MCN-3, but not limited to this.Preferably, it is given an account of
Hole carbon can select MCN-1 or MCN-3.More preferably, the mesoporous carbon can select MCN-3.
It should be noted that the present invention passes through specific surface area (SBET), Kong Rong (VM) and pore-size distribution (Dm) define MCN-
1, MCN-2 and MCN-3.
Wherein, it is 755-765m that MCN-1, which refers to specific surface area,2/ g, Kong Rongwei 0.8-0.95cm3/ g, pore-size distribution 3.9-
The mesoporous carbon of 7.8nm；
MCN-2 refers to that specific surface area is 740-754m2/ g, Kong Rongwei 0.96-1.15cm3/ g, pore-size distribution 3.9-7.6
The mesoporous carbon of nm；
MCN-3 refers to that specific surface area is 480-530m2/ g, Kong Rongwei 0.50-0.80cm3/ g, pore-size distribution 8.0-
The mesoporous carbon of 20nm.
Correspondingly, the present invention discloses a kind of preparation method such as the mesoporous carbon particle of insulin, including:
10-15mg mesoporous carbons and 3-6mg insulin are placed in test tube, the alcoholic solution and 0.1- of 0.4-1.2mL is added
The acid solution of 0.4mL, a concentration of 0.005-0.03mol/L of acid solution, vortex oscillation 20-50min；
Sample after oscillation is transferred in evaporating dish, and room temperature drying obtains the mesoporous carbon particle of insulin.
The preparation method of the mesoporous carbon particle of insulin of the present invention is simple, only includes two steps, and alcoholic solution and acid are molten
Liquid is added simultaneously inside a step, simplifies operating procedure, saves preparation time.And documents 1 need by
Insulin is first dissolved in hydrochloric acid, adds silica, dry after insulin and aerosil absorption completely, obtains
Solid；Then PEG is dissolved in absolute ethyl alcohol, then solid is dissolved in ethanol solution, final emulsion drying obtains particle.And
And aerosil have hydrophobicity when, need first to make its tool through 300~1000 DEG C of heat treatments before hydrochloric acid solution is added
There is hydrophily.
Preferably, the dosage of the mesoporous carbon is 11-13mg, and the dosage of the insulin is 4-5mg, the alcoholic solution
Dosage is 0.6-1.0mL, and the dosage of the acid solution is 0.2-0.3mL, a concentration of 0.01-0.02mol/L of acid solution, whirlpool
The time of rotation oscillation is 25-35min.It is hydrochloric acid or acetic acid that the alcoholic solution, which selects methanol or ethyl alcohol, the acid solution,.
More preferably, the dosage of the mesoporous carbon is 12mg, and the dosage of the insulin is 4mg, the dosage of the alcoholic solution
For 0.8mL, the dosage of the acid solution is 0.2mL, a concentration of 0.01mol/L of acid solution, and the time of vortex oscillation is
30min.It is acetic acid that the alcoholic solution, which selects methanol, the acid solution,.
Correspondingly, the present invention also provides the insulin oral preparations made of above-mentioned insulin mesoporous carbon particle.It is described
Oral preparation is pharmaceutically acceptable oral preparation, including tablet, pill, powder, capsule, granule or suspension.Together
When, the mesoporous carbon particle of insulin of the present invention is also possible to be made into pharmaceutically acceptable injection or suppository.
Correspondingly, the application the present invention also provides the mesoporous carbon particle of insulin in treating diabetes.
Three kinds of meso-porous carbon materials MCN-1, MCN-2 that the present invention is provided with pharmacy teaching and research room of Guangdong pharmaceutical university and
MCN-3 is raw material, by the modes such as stir, volatilize synthesize the different insulin mesoporous carbon nanoparticle MCNI-1 of three kinds of properties,
MCNI-2 and MCNI-3 is surveyed by scanning electron microscope collection of illustrative plates, transmission electron microscope collection of illustrative plates, infared spectrum and specific surface area, pore-size distribution
It is fixed that comparison characterization is carried out to blank meso-porous carbon material and the insulin mesoporous carbon nanoparticle synthesized, and by measuring mesoporous carbon materials
The insulin concentration after being adsorbed to insulin solutions in filtrate is expected to calculate drugloading rate of the meso-porous carbon material to insulin, and is ground
Release behavior of the insulin in the medium of simulation intestinal environment is studied carefully.The last present invention carries out insulin mesoporous carbon nanoparticle pair
Adsorption test of the oral pharmacodynamic study and FITC fluorescent markers meso-porous carbon material of diabetes rat to HCT-116 cells.
The result shows that the scanning electron microscope collection of illustrative plates of blank meso-porous carbon material and insulin mesoporous carbon oral administration nanometer grain and transmission electricity
Mirror collection of illustrative plates has significant difference, the infared spectrum of insulin mesoporous carbon nanoparticle to show that the characteristic peak of insulin, insulin are mesoporous
The specific surface area of carbon nanoparticle is significantly reduced than blank meso-porous carbon material, pore-size distribution significant changes.Adsorption rate and drugloading rate are surveyed
Take temperature bright, the adsorption rate of MCNI-1, MCNI-2 and MCNI-3 be respectively 41.52% ± 1.99%, 28.77% ± 0.30% and
49.49% ± 1.78%, drugloading rate is respectively 12.16% ± 0.51%, 8.68% ± 0.08% and 14.06% ± 0.43%,
Meet the pharmacodynamics demand of this experiment.In addition in releasing research of the insulin mesoporous carbon nanoparticle in pH7.4PBS solution,
The release of MCNI-1, MCNI-2 and MCNI-3 in 15min can reach 48.20% ± 8.93%, 15.68% respectively ±
5.50% and 53.11% ± 3.69%, realize being released effectively in simulated intestinal fluid environment.Insulin mesoporous carbon nanoparticle presses pancreas
Island element 50IU/kg dosage to the oral pharmacodynamics of diabetes rat experiments have shown that, blood glucose in diabetic rats is aobvious after 4h is administered orally
Writing reduces, and has the effect of sustained release compared with being subcutaneously injected, wherein MCNI-1, MCNI-2 and MCNI-3 are to blood glucose in diabetic rats
Most low energy degrades 13.2% ± 5.2%, 62.3% ± 17.0% and the 48.7% ± 21.3% of original initial blood glucose.FITC labels are situated between
Hole carbon shows that meso-porous carbon material used herein has HCT-116 cells certain absorption to HCT-116 adsorption tests
Ability, the transporting mechanism research for later insulin mesoporous carbon nanoparticle in intestinal cell provide the foundation.
It is detailed content of the test below.
1. the physicochemical property of insulin mesoporous carbon nanoparticle characterizes
It takes each 12mg of meso-porous carbon material MCN-1, MCN-2 and MCN-3 to be placed in 1mL EP pipes, is separately added into 4mg insulin,
0.8mL methanol and 0.2mL 0.01mol/L HCl, vortex oscillation 30min is added.The sample that oscillation finishes is transferred to steaming later
Send out in ware, room temperature drying, obtain for scanning electron microscope collection of illustrative plates insulin mesoporous carbon nanoparticle MCNI-1, MCNI-2 for using of test and
MCNI-3.In addition, preparing each 12mg of blank meso-porous carbon material MCN-1, MCN-2 and MCN-3.Scanning electron microscope diagram spectrum is surveyed
Strip part：SEM and efficient energy disperse spectroscopy (Energy dispersive spectroscopy, EDS) are Germany Merlin
Emission electron microscope, accelerating potential 5kV.Transmission electron microscope figure composes test condition：200kV
(Japanese JEOL 2100F).Sample powder is done in alcohol ultrasonic disperse by hanging drop on the copper mesh rich in porous carbon membrane
Dry post analysis.The specific surface area of insulin mesoporous carbon nanoparticle and blank meso-porous carbon material is by Brunauer-Emmett-
Teller (BET) method is measured, and the computational methods of Kong Rong and pore-size distribution are Barrett-Joyner-Halenda (BJH) mould
Type.
Take each 2mg of insulin mesoporous carbon nanoparticle MCNI-1, MCNI-2 and MCNI-3 obtained above, respectively with 200mg
Dry KBr powder is uniform with agate mortar drying and grinding under infrared lamp, and thin slice is made with tablet press machine, is placed in ir spectrophotometry
It is detected in meter, scanning range：4000cm-1~500cm-1, scanning times：32 times, record gained collection of illustrative plates.
Fig. 1 is the SEM spectrum (A.MCN-1 of blank mesoporous carbon sample and insulin mesoporous carbon sample；B. MCNI-1；
C.MCN-2；D.MCNI-2；E.MCN-3；F.MCNI-3), it is known that, insulin mesoporous carbon nanoparticle and blank meso-porous carbon material phase
Than, the mesoporous carbon surface of blank and duct edge clear, uniformly；And the surface of insulin mesoporous carbon nanoparticle is covered by insulin medicament
Lid, causes meso-porous carbon material surface and its duct edge to fog.
Fig. 2 is the TEM collection of illustrative plates (A.MCN-1 of blank mesoporous carbon sample and insulin mesoporous carbon sample；B. MCNI-1；
C.MCN-2；D.MCNI-2；E.MCN-3；F.MCNI-3), the duct of blank meso-porous carbon material is clear under transmission electron microscope
As it can be seen that but the duct of insulin mesoporous carbon nanoparticle that has synthesized cannot differentiate, show that insulin molecule has been integrated into
The duct of mesoporous carbon, and then realize and carry medicine, success insulin synthesis mesoporous carbon nanoparticle.
Fig. 3 is the FTIR spectrum (A. of insulin, blank mesoporous carbon sample and insulin mesoporous carbon nanoparticle
Insulin；B.MCN-1；C.MCNI-1；D.MCN-2；E.MCNI-2；F.MCN-3；G.MCNI-3), due to the mesoporous carbon materials of blank
Material unavoidably generation-C=O, therefore the peaks-the C=O (~1616.06cm in blank meso-porous carbon material in sintering procedure-1)
It can overlap with the peaks-C=O of insulin.And in 4000~3000cm-1In wave-number range ,-the NH- ,-NH of insulin2、-OH
And the peak of C ≡ C or the C-H structures of the peaks-COOH and mesoporous carbon overlaps and results in the variation of peak type, or even there are conjugations
Lead to the appearance of broad peak.In 2000~1000 cm-1In wave-number range ,-the NH- ,-NH of insulin molecule2, the peaks such as-OH are in pancreas islet
There is shown, the synthesis success of expression insulin mesoporous carbon nanoparticle in plain mesoporous carbon nanoparticle.
Table 1 is the specific surface area (S of blank mesoporous carbon sample and insulin mesoporous carbon nanoparticleBET), Kong Rong (VM) and hole
Diameter is distributed (Dm)：
Table 1
The specific surface area and Kong Rongjun of each model insulin mesoporous carbon nanoparticle are than original corresponding blank meso-porous carbon material
It reduces, wherein SBETThe ratio MCNI-1 of reduction>MCNI-3>MCNI-2, VMThe ratio MCNI-1 of reduction> MCNI-2>MCNI-3.
On the pore-size distribution that BJH models obtain, MCNI-1 does not have significant change, MCNI-2 to compare compared with MCN-2 and then lose MCN-1
The pore-size distribution peak of 3.9nm, MCNI-3 are reduced to 6.8nm compared to pore-size distribution peak compared with MCN-3 by 8.5nm.
2. meso-porous carbon material measures insulin adsorption rate, drugloading rate
By measuring meso-porous carbon material to the concentration of the remaining insulin of filtrate after insulin absorption in solution, calculate indirectly
Adsorption rate and drugloading rate of the mesoporous carbon to insulin.The computational methods of wherein adsorption rate and drugloading rate are：
This experiment is measured using high performance liquid chromatography (HPLC), is pressed《Chinese Pharmacopoeia》Under the two pancreas islet prime implicants of version in 2015
It measures, chromatographic condition is：
Mobile phase：0.2mol/L sulfate buffers (anhydrous sodium sulfate containing 28.4g and 2.7ml phosphoric acid, with ethanol amine tune pH
To 2.3)-acetonitrile (74:26), flow velocity 1ml/min；Chromatographic column：C18 (4.6 × 150mm, 3 μm)；Detector：D2 (λ=
214nm)；Appearance time：About 10min.
The meso-porous carbon material (being respectively MCN-1, MCN-2 and MCN-3) of 4.0mg insulin and 12.0mg is taken to be placed in 10ml
In volumetric flask, PBS solution is added to be settled to scale, place it in ice bath makes mesoporous carbon and pancreas islet with 150rpm rotating speed shaking tables 12h
Plain fully absorption.It takes its filtrate to survey the insulin peak area of each filtrate through HPLC methods, according to standard curve, calculates the pancreas in solution
Island element residual concentration calculates adsorption rate and drugloading rate of each meso-porous carbon material to insulin in solution indirectly.
Table 2 is the adsorption rate and drugloading rate of meso-porous carbon material MCN-1, MCN-2 and MCN-3 to insulin,
Table 2
As shown in Table 2, three kinds of meso-porous carbon materials have insulin apparent suction-operated in experiment.
3. the release performance of insulin mesoporous carbon nanoparticle is studied
Release performance research, as dissolution medium, it is mesoporous to detect obtained insulin using 37 DEG C of pH7.4PBS solution
Release performance of the carbon nanoparticle in the case where simulating colonic environment, dosage foundation is provided for next step pharmacodynamics test.This was tested
Per the dissolution medium required supplementation with after sub-sampling with sampling same volume in journey, thus during this experiment release calculating
Method is：
Wherein：
cnThe concentration (mg/mL) of insulin in-each sample point filtrate
cmaxThe concentration that filtrate should reach when-insulin mesoporous carbon sample theoretically discharges completely in dissolution medium
(mg/mL)
The sample volume (mL) at V-each time points
VmaxThe total volume (mL) of-dissolution medium
This experiment is measured using high performance liquid chromatography (HPLC), is pressed《Chinese Pharmacopoeia》Under the two pancreas islet prime implicants of version in 2015
It measures, chromatographic condition is as hereinbefore.
Prepare three 100mL round-bottomed flasks, each pH7.4PBS solution for being packed into 50mL is placed in electric heating magnetic as dissolution medium
On power blender, if 37 DEG C of water-bath, stirred with 300rpm rotating speeds.Weigh the insulin mesoporous carbon nanoparticle of insulin-containing 5.0mg
When the temperature of medium to be released maintains 37 DEG C, above-mentioned insulin mesoporous carbon nanoparticle is added by MCNI-1, MCNI-2 and MCNI-3
Enter in dissolution medium, and start timing, sampled by 0.2mL every 15min, in 30s in filtering to 1mL EP pipes, sampling is backward
0.2mL pH7.4PBS solution is supplemented in delivery systme, this experiment is maintained to 120min.The filtrate that each time point is acquired, warp
HPLC methods detect the concentration of insulin in each filtrate, and insulin mesoporous carbon nanoparticle MCNI-1, MCNI- are calculated according to standard curve
2 and MCNI-3 characterizes its release performance in the insulin releasing degree at each time point.
Table 3 and Fig. 4 are release of the insulin mesoporous carbon nanoparticle in pH7.4PBS solution,
Table 3
*P ＜ 0.001vs MCNI-1 or MCNI-3
By watch 3 and Fig. 4 it is found that insulin mesoporous carbon sample can realize fast quick-release in pH7.4PBS solution in 15min
It puts, the insulin releasing rate of MCNI-1, MCNI-2 and MCNI-3 respectively reach 48.20% ± 8.93%, 15.68% when 15min
± 5.50% and 53.11% ± 3.69%.
Hypoglycemic influence of the 4 oral insulin mesoporous carbon nanoparticles to diabetes rat
Pharmacodynamics test, as experimental animal, passes through oral insulin mesoporous carbon nanoparticle using diabetic model rats
Diabetic model rats are administered in mode, and the change of blood sugar of each rat, preliminary proof oral insulin mesoporous carbon are measured every 2h
The mode of nanoparticle can reduce the blood glucose of diabetes rat.
The foundation of the preparation and diabetic model rats of 4.1 streptozotocin injections
Streptozotocin can establish rat diabetes model, but chain urea destroying in rat body by way of B cell
Help rhzomorph high temperature, light-exposed easy decomposition, it is therefore necessary to now with the current.In order to make streptozotocin stability of solution improve, this experiment
Streptozotocin injection is prepared as solvent using 0.1mol/L pH4.5 citric acid-sodium citrate buffer solutions, in injection
A concentration of 5mg/mL of streptozotocin.
Select Healthy female rat several, the streptozotocin that above-mentioned preparation is injected intraperitoneally by 50mg/kg body weight doses is injected
Liquid carries out second of injection after 7d again.A large amount of discharge into blood is caused because administration initial stage B cell is destroyed rear insulin
It crosses hypoglycemia and dead, is administered after 2h to need to give 5% glucose solution to rat and drink to maintain blood glucose.Away from second of injection 7d
Afterwards, to each rat limosis 6h, its tail vein blood glucose is surveyed with blood glucose meter, rat of the blood glucose more than 11.1mmol/L is diabetes model
The rat being successfully established.
Hypoglycemic effect of the 4.2 oral insulin mesoporous carbon nanoparticles to diabetes rat
Three 50mL centrifuge tubes are taken, load MCN-1, MCN-2 and MCN-3 of 74.0mg respectively, insulin is then respectively added
3.7mg (is equivalent to 100IU).1.0mL 0.01mol/L HCl, which are added, makes insulin dissolve, and adds 19.0mL pH7.4PBS
Solution seals vortex oscillation 30min, and 4 DEG C stand overnight, and obtain the insulin mesoporous carbon nanoparticles oral suspension of 5IU/mL,
As test group.
A 50mL centrifuge tube is taken, 3.7mg insulin (being equivalent to 100IU) is added, 0.01 mol/L of 1.0mL are added
HCl makes insulin dissolve, and adds 19.0mL pH7.4PBS solution, shakes up, and 4 DEG C stand overnight, obtains the insulin of 5IU/mL
Oral administration solution, as blank control group.
A 50mL centrifuge tube is taken, 0.37mg insulin (being equivalent to 10IU) is added, 0.01 mol/L of 1.0mL are added
HCl makes insulin dissolve, and adds 19.0mL pH7.4PBS solution, shakes up, and 4 DEG C stand overnight, obtains the pancreas islet of 0.5IU/mL
Liquid is subcutaneously injected in element, as positive controls.
Every group of 10 rats are randomly assigned, empty stomach 2h before testing, wherein the hypodermic injection group as positive control is to medicament
Amount is 5IU/kg, and the insulin gavage group as blank control and the insulin mesoporous carbon nanoparticle gavage group as test group are given
Pharmaceutical quantities are 50IU/kg, and administration measures the blood sugar concentration of each rat every 2h with blood glucose meter after starting, and with initial blood glucose (0h)
Compare, calculate the percentage of the blood glucose and initial blood glucose at each time point, draws change of blood sugar curve.
Fig. 5 is the percent change value of blood glucose after Oral Administration in Rats insulin mesoporous carbon nanoparticle, positive controls (subcutaneous note
Penetrate 5IU/kg groups) upon administration 2h reach blood glucose minimum, be initial blood glucose 16.0% ± 2.9%, subsequent blood glucose gradually returns
It rises, 8h returns to initial blood glucose (92.9% ± 7.8%) after administration.MCNI-1 takes orally group (50IU/kg) 4h after administration and reaches blood glucose
Minimum (13.2% ± 5.2%), 12h only returns to the 70.3% ± 29.6% of initial blood glucose after administration.MCNI-2 takes orally group
(50IU/kg) 4h after administration reaches blood glucose minimum (62.3% ± 17.0%), and 10h returns to initial blood glucose after administration
(104.1% ± 20.7%).MCNI-3 take orally group (50 IU/kg) after administration 6h up to blood glucose minimum (48.7% ±
21.3%), 8h returns to initial blood glucose (100.5% ± 2.9%) after administration.
Adsorption test of 5 meso-porous carbon materials to colon cancer cell HCT-116
By meso-porous carbon material MCN-1, MCN-2 and MCN-3 of FITC fluorescent markers to colon cancer HCT-116 cells
Absorption property of the above-mentioned meso-porous carbon material to HCT-116 cells is investigated in adsorption test, is existed for later insulin mesoporous carbon nanoparticle
The transporting mechanism research of colonic environment provides basis.
The synthesis of 5.1FITC fluorescent marker meso-porous carbon materials
Three 100mL round-bottomed flasks are taken, are protected from light with aluminium foil package, every each addition 20mL ethyl alcohol, 10mg FITC powder
And 100 μ L, stirring 8h is protected from light at 25 DEG C, obtains reaction solution.At the same time, prepare three 15mL centrifuge tubes, weigh respectively MCN-1,
Each 20mg of MCN-2 and MCN-3, are then respectively adding 30% hydrogen peroxide of 5mL by mesoporous carbon surface oxidation, for when 8h.Later will
Meso-porous carbon material after oxidation is washed with 0.22 μm of filtering with microporous membrane and with ethyl alcohol respectively, is then mixed with 10mL ethyl alcohol
It is outstanding, it is added in above-mentioned reaction solution and continues to stir 12h.Products therefrom will then be synthesized and be transferred to the 50mL centrifuge tubes that aluminium foil wraps up
In, 5min is centrifuged with 3000rpm rotating speeds, discards supernatant liquid, ethyl alcohol 10mL is added and is suspended, washs, then 3000rpm centrifuges 5min,
Liquid is discarded supernatant, repeats and washes twice, obtains the meso-porous carbon material of FITC fluorescent markers.
Adsorption test of 5.2 meso-porous carbon materials to colon cancer cell HCT-116
By DMEM culture medium (hereinafter referred to as DMEM+10%FBS of the HCT-116 cells containing 10% fetal calf serum (FBS)
Culture medium) it is diluted to 5 × 104mL-1, and be seeded in 13 45cm culture dishes, culture is for 24 hours.By synthesized FITC fluorescence marks
Note meso-porous carbon material is added 75% ethyl alcohol of 5mL and shakes up, sterilizes, and 2000 rpm centrifuge 4min, discard supernatant liquid, and 5mL is added and goes out
Bacterium PBS shakes up, washs, and 2000rpm centrifuges 4 min, discards supernatant liquid, then washed once with sterilizing PBS.After discarding supernatant liquid,
10mL DMEM+10%FBS culture mediums, which are added, makes a concentration of 2mg/mL of FITC fluorescent marker mesoporous carbons.
13 culture dishes are divided into three groups, every group four, remaining one is used as blank control group.By the training in each culture dish
Foster base discards, and new culture medium is added in blank control group, remaining each group is separately added into each FITC fluorescent markers meso-porous carbon material
Each 2mL of suspension.It is protected from light culture, the suspension on cell is discarded away from administration 0.5h, 1h, 2h and 4h, PBS solution is used in combination to wash three
It is secondary, cell is fixed with methanol, to observe the FITC that HCT-116 is adsorbed under 488nm excitation wavelengths under fluorescence inverted microscope
Mark mesoporous carbon particle, amplification factor 100 ×, and record its fluorescence micrograph.
After Fig. 6 shows that the meso-porous carbon material of FITC fluorescent markers acts on colon cancer HCT-116 cells 0.5h, three kinds of Jie
Hole carbon material is adsorbed with HCT-116 cells, and delaying with the time, and the mesoporous carbon for participating in absorption is more.This experiment is indirect
Show that three kinds of mesoporous carbons there is certain adsorption capacity, insulin mesoporous carbon nanoparticle can be realized in intestinal cell colon cell
Transhipment.
It is above disclosed to be only a preferred embodiment of the present invention, the power of the present invention cannot be limited with this certainly
Sharp range, therefore equivalent changes made in accordance with the claims of the present invention, are still within the scope of the present invention.
Claims (10)
1. a kind of mesoporous carbon particle of insulin, which is characterized in that including mesoporous carbon and insulin, the insulin is embedded in mesoporous
In carbon, the amount ratio of the insulin and mesoporous carbon is 3-6:10-15.
2. the mesoporous carbon particle of insulin as described in claim 1, which is characterized in that the amount ratio of the insulin and mesoporous carbon
For 4-5:11-13.
3. the mesoporous carbon particle of insulin as described in claim 1, which is characterized in that the grain size of the mesoporous carbon be 2nm~
50nm。
4. the mesoporous carbon particle of insulin as claimed in claim 3, which is characterized in that the grain size of the mesoporous carbon be 10nm~
40nm。
5. the mesoporous carbon particle of insulin according to any one of claims 1-4, which is characterized in that the mesoporous carbon selects MCN-
1, MCN-2 or MCN-3, wherein
MCN-1 refers to that specific surface area is 755-765m2/ g, Kong Rongwei 0.8-0.95cm3/ g, pore-size distribution are Jie of 3.9-7.8nm
Hole carbon；
MCN-2 refers to that specific surface area is 740-754m2/ g, Kong Rongwei 0.96-1.15cm3/ g, pore-size distribution are 3.9-7.6nm's
Mesoporous carbon；
MCN-3 refers to that specific surface area is 480-530m2/ g, Kong Rongwei 0.50-0.80cm3/ g, pore-size distribution are Jie of 8.0-20nm
Hole carbon.
6. a kind of preparation method of the mesoporous carbon particle of insulin as described in any one in claim 1-5, which is characterized in that packet
It includes:
10-15mg mesoporous carbons and 3-6mg insulin be placed in test tube, the alcoholic solution and 0.1-0.4mL of 0.4-1.2mL is added
Acid solution, a concentration of 0.005-0.03mol/L of acid solution, vortex oscillation 20-50min；
Sample after oscillation is transferred in evaporating dish, and room temperature drying obtains the mesoporous carbon particle of insulin.
7. the preparation method of the mesoporous carbon particle of insulin as claimed in claim 6, which is characterized in that the dosage of the mesoporous carbon
Dosage for 11-13mg, the insulin is 4-5mg, and the dosage of the alcoholic solution is 0.6-1.0mL, the use of the acid solution
Amount is 0.2-0.3mL, and the time of a concentration of 0.01-0.02mol/L of acid solution, vortex oscillation are 25-35min.
8. the preparation method of the mesoporous carbon particle of insulin as claimed in claims 6 or 7, which is characterized in that the alcoholic solution choosing
With methanol or ethyl alcohol, the acid solution is hydrochloric acid or acetic acid.
9. a kind of insulin oral preparation, which is characterized in that it includes that insulin as described in any one in claim 1-5 is mesoporous
Carbon particle.
10. a kind of application of mesoporous carbon particle of insulin as described in any one in claim 1-5 in treating diabetes.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103769052A (en) * | 2014-02-28 | 2014-05-07 | 广东药学院 | Magnetism and temperature double-responded mesoporous carbon material as well as preparation method and application thereof |
| CN103990133A (en) * | 2014-05-07 | 2014-08-20 | 沈阳药科大学 | Mesoporous carbon nanoparticle system with targeting site-specific drug release and application thereof |
| CN104940141A (en) * | 2015-07-10 | 2015-09-30 | 东北林业大学 | Preparation method for oral insulin enteric-coated preparations |
| 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 |
-
2017
- 2017-12-29 CN CN201711478291.5A patent/CN108421049B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103769052A (en) * | 2014-02-28 | 2014-05-07 | 广东药学院 | Magnetism and temperature double-responded mesoporous carbon material as well as preparation method and application thereof |
| CN103990133A (en) * | 2014-05-07 | 2014-08-20 | 沈阳药科大学 | Mesoporous carbon nanoparticle system with targeting site-specific drug release and application thereof |
| CN104940141A (en) * | 2015-07-10 | 2015-09-30 | 东北林业大学 | Preparation method for oral insulin enteric-coated preparations |
| 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 |
Non-Patent Citations (5)
| Title |
|---|
| ABDOLLAH ZAKERI SIAVASHANI ET AL: ""Preparation of mesoporous silica nanoparticles for insulin drug delivery"", 《ADVANCED MATERIALS RESEARCH》 * |
| QINFU ZHAO ET AL: ""Mesoporous carbon nanomaterials in drug delivery and biomedical application"", 《DRUG DELIVERY》 * |
| XIAOJING ZHU ET AL: ""Controllable synthesis of mesoporous carbon nanospheres with uniform size by a facile one-pot aqueous strategy under highly acidic conditions"", 《CARBON》 * |
| 林浩荣: "胰岛素介孔碳纳米粒的制备及其口服药效学评价", 《万方数据库》 * |
| 钟国英，等: "新型有序介孔材料在药物传递系统中的应用", 《材料导报：综述篇》 * |
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