CN108273042A - A kind of ginsenoside-insulin nano gel and the preparation method and application thereof - Google Patents
A kind of ginsenoside-insulin nano gel and the preparation method and application thereof Download PDFInfo
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- 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
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
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- 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
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Abstract
The invention discloses a kind of ginsenoside insulin nano gels and the preparation method and application thereof, it is using hypoglycemic drug insulin as effective component, in conjunction with the ginsenoside gel nano particles with effect of lowering blood sugar, a kind of ginsenoside insulin nano gel with dual hypoglycemic effect is prepared；Wherein, the percutaneous sustained-release administration of insulin can be achieved in the use of ginsenoside gel nano particles, avoids the side effect that injection of insulin is brought.Ginsenoside insulin nano gel of the present invention is used to prepare percutaneous drug administration preparation, has the characteristics that percutaneous absorbtion rate fast, safety and stability and painless convenient, and preparation method is simple, production cost is low, and technique is easy to amplify.
Description
Technical field
The invention belongs to pharmaceutical preparation technical fields, and in particular to a kind of ginsenoside-insulin nano gel and its system
Preparation Method and application.
Background technology
Diabetes are a kind of metabolic diseases characterized by hyperglycemia, it has also become after cardiovascular and cerebrovascular disease and tumour
It endangers " the third killer " of human health.According to International Diabetes Federation（IDF）Report, global diabetic in 2017
4.25 hundred million people, wherein diabetes mellitus in China number of patients are up to 1.144 hundred million people, and number of patients height ranks first in the world, pre- to the year two thousand forty
Meter will rise to 1.507 hundred million people.Once blood glucose controls bad for a long time for diabetic, it is likely that can cause it is serious simultaneously
Disease is sent out, such as diabetic retinopathy will cause blindness, diabetic nephropathy that will lead to Chronic renal failure simultaneously
Send out disease.China has 1,300,000 people to die of diabetes and its complication within 2017, and therefore, treating diabetes are very urgent.
Two kinds of approach of the universally recognized treatment diabetes of medical field are mainly oral drug therapy and insulin therapy,
Middle insulin accounts for more than half of whole market.Insulin（insulin）Be pancreatic beta cell secrete, it is unique it is a kind of can be
The hormone of blood glucose and the first-line drug of diabetes clinic are reduced in vivo.Whole type-1 diabetes mellitus people and after the II types at present
The diabetes patient of phase must receive injection of insulin treatment.But long term injections insulin also brings along problems, such as patient
It can be by huge psychology and physiology pain；Long term injections can cause the formation of injection site fat pad, this fat pad to be deposited
The absorption of local insulin can be being seriously affected, insulin resistance is occurring, drug effect is caused to reduce；Meanwhile it there is also such as water
The symptoms such as swollen, part silt blueness, skin infection, instantaneous hypoglycemia；And it is expensive, need to frequently inject.These are all current skills
The inevitable problem of insulin injection under the conditions of art.
Transdermal delivery system（Transdermal drug delivery system, TDDS）Refer to drug with certain speed
Rate enters a kind of preparation of body circulation through skin through capillary absorbance.TDDS can realize that non-invasive is administered, and have and surmount
The particular advantages of general medication：It directly acts on target area and plays drug effect；Avoid the first pass effect and gastrointestinal tract of liver
Interference；Blood concentration is stable, reduces administration number of times etc..But percutaneous dosing still has limitation, i.e. bioprotein class macromolecular,
Such as insulin molecule, it is difficult to which passing freely through keratoderma realizes percutaneous dosing.
The present invention uses nanotechnology, and by insulin and ginsenoside self assembly, a kind of percutaneous drug administration preparation, tool is made
There is smaller nano-scale, the absorption and infiltration of drug can be promoted.Skin is acted on, insulin can be solidifying from ginsenoside
Stablize release in gel matrix, in entering in vivo through Cutaneous permeation, plays hypoglycemic effect.
Invention content
For reduce side effect that existing injection of insulin agent brings, solve physiology that injection of insulin agent is brought to patient and
Mental anguish and the problems such as it is expensive, the present invention provides a kind of ginsenoside-insulin nano gel and its preparation sides
Method and application have the advantages that the drug effect phase is long and economical convenient.
To achieve the above object, the present invention adopts the following technical scheme that：
A kind of ginsenoside-insulin nano gel is to be self-assembly of ginsenoside gel nano particles with insulin
Ginsenoside/insulin gel nano particles, then itself and matrix, moisturizer, penetration-assisting agent are mixed to prepare the ginsenoside-pancreas
Island element nanogel.
The preparation of the ginsenoside gel nano particles includes the following steps：
1）With one or more extractions in Araliaceae China ginseng, Korean ginseng, American Ginseng, Vietnam's ginseng, Japanese ginseng
General ginsenoside be raw material, in acidic aqueous solution carry out acidolysis reaction obtain ginsenoside acid hydrolysis solution；
2）By step 1）After the purified removal impurity of gained ginsenoside acid hydrolysis solution, it is re-dissolved in water to obtain the final product.
Step 1）Described in acidic aqueous solution be organic and or inorganic acids aqueous solution, pH value is<7, most preferably 1-
3；The temperature of the acidolysis reaction is 50 ~ 105 DEG C；Time is 1 ~ 72h.
Step 2）Concrete operations be：Ginsenoside acid hydrolysis solution is stood 2 hours in -25 DEG C~35 DEG C or more, then pass through
Centrifugation or filtering removal precipitation, adjust pH to 7.5~14 by clear liquid organic base and/or inorganic base, obtain sediment；At 25 DEG C
At~85 DEG C, sediment is dissolved in organic solvent, solution is then cooled to -20 DEG C~10 DEG C, through centrifuging or filtering off
Except being dried after precipitation；
The organic solvent is dimethyl sulfoxide (DMSO), methanol, ethyl alcohol, n-butanol, propyl alcohol, tetrahydrofuran, one kind in pyridine or more
Kind；The drying is constant pressure and dry, dry or spray drying is concentrated under reduced pressure.
The average grain diameter of the ginsenoside/insulin gel nano particles is 30 ~ 100nm, and the grain size of particle is smaller, warp
The efficiency that skin absorbs is higher.
The matrix includes natural polymer（Such as starch, alginate, tragacanth, agar and gelatin）, semi-synthetic height
Molecule（Such as modified starch and modified cellulose, carboxymethyl cellulose, methylcellulose）With synthesis macromolecule（Such as carbomer gathers
Sodium acrylate）In it is one or more；The moisturizer is glycerine, trimethylglycine, saualane, cyclopolysiloxane, tristearin
It is one or more in acid, cholesterol, lecithin etc.；The penetration-assisting agent be propylene glycol, azone, pyrrolones, aliphatic acid and its
It is one or more in esters, cyclodextrin, sulfoxide type etc..
The preparation method of the ginsenoside-insulin nano gel includes the following steps：
1）Insulin and ginsenoside gel nano particles are taken, secondary water is added, makes ginsenoside gel nano particles in solution
A concentration of 1 ~ 40 mg/mL, insulin a concentration of 1 ~ 25U/mL, ultrasonic vibration 15min is solidifying to get ginsenoside/insulin
Glue nanoparticles solution；
2）Matrix, moisturizer and penetration-assisting agent are dissolved in secondary water, ginsenoside/insulin gel is added under stirring conditions
Nanoparticles solution, up to ginsenoside-insulin nano gel after mixing well.
The ginsenoside-insulin nano gel is used to prepare answering in the percutaneous drug administration preparation with function of reducing blood sugar
With, it is characterised in that：The percutaneous drug administration preparation includes ointment, patch, spray.
At least contain 1.6mg/cm in the percutaneous drug administration preparation2Insulin, 40mg/cm2Ginsenoside gel nanometer
Grain.
Present invention has the advantage that：
（1）Ginsenoside of the present invention-insulin nano gel has the same of hypoglycemic effect using insulin, ginsenoside
When, promote insulin to penetrate into vivo using the ginsenoside for being prepared into gel nano particles, and reach good dual drop blood
The effect of sugar and medicament slow release.
（2）Ginsenoside of the present invention-insulin nano gel has extra small grain size, the potential of stabilization and higher load medicine
Amount, and blood vessel is penetrated readily through without causing blood vessel endothelium injury, enzyme degradation can be protected a drug from.By ginsenoside-of the present invention
Drug in percutaneous drug administration preparation prepared by insulin nano gel discharges 80% in 2 days, and unit area adds up transmitance and reaches
0.31mg/cm2。
（3）Hypoglycemic treatment is carried out using percutaneous drug administration preparation prepared by ginsenoside of the present invention-insulin nano gel,
The fat pad phenomenon caused by hypodermic injection can be avoided, to avoid its influence to drug absorption, and can effectively be kept away
Instantaneous hypoglycemia caused by possibility when exempting to be subcutaneously injected.
（4）The present invention can under conditions of not by complicated approach such as electro-ionic osmosis, ultrasonic wave, electroporation and electrets and
The percutaneous dosing for realizing insulin, the research and development for insulin percutaneous dosing is provided fundamental basis and researching value.
Description of the drawings
Fig. 1 is the grain size of ginsenoside-insulin nano gel（A）And potential energy diagram（B）.
Fig. 2 is that the atom of ginsenoside-insulin nano gel is tried hard to.
Fig. 3 is the standard curve that BCA methods survey protein quantification.
Fig. 4 is that the cumulative release amount of insulin in ginsenoside-insulin nano gel changes over time figure.
Fig. 5 is the cumulative in vitro transit dose that the outer percutaneous absorbtion of Mice Body tests insulin.
Fig. 6 is the blood glucose value variation of different time points in hyperglycemia Mice Body.
Specific implementation mode
In order to make content of the present invention easily facilitate understanding, With reference to embodiment to of the present invention
Technical solution is described further, but the present invention is not limited only to this.
Embodiment 1：
1) 5g general ginsenosides are weighed, the acetum of pH 1.8 is added to 25ml, 4h, gained reactant are decomposed at 90 DEG C
It is placed at room temperature for after 12h with 0.45 μm of membrane filtration, removes insoluble matter, it is 8 that filtrate is neutralized to pH with 10% sodium carbonate, then with 2 μm
Membrane filtration collects precipitation；Sediment is added in the absolute ethyl alcohol of 20 mL, is heated to 60 DEG C and makes it dissolve, 4 DEG C of decentralizations
It is filtered with 2 μm of filter paper after setting cooling 2h, ginsenoside gel powder is drying to obtain after filtrate decompression concentration；
2）Insulin and ginsenoside gel powder are taken, secondary water is added, makes final concentration of 1 ~ 40 mg/ of ginsenoside in solution
A concentration of 1 ~ 25U/mL of mL, insulin, ultrasonic vibration 15min are molten to get ginsenoside/insulin nano gel nano particles
Liquid；
3）0.5g carbomers, 0.35g gelatin, 0.5mg glycerine, 0.1ml propylene glycol are dissolved in 2ml secondary waters, in the item of stirring
Ginsenoside/insulin gel nano particles solution obtained by 2ml is added under part, is uniformly mixed and is received to get ginsenoside/insulin
Rice gel.
The characterization of 2 ginsenosides of embodiment-insulin nano gel：
1）The grain size (size, nm) and potential (zeta potential, mv) of nanogel are measured with dynamic light scattering (DLS),
The results are shown in Figure 1.
As seen from Figure 1, the nanogel that prepared by the present invention has small grain size and negative potential.
2）The three-dimensional structure of nanogel is measured with atomic force microscope (AFM), the results are shown in Figure 2.
From Figure 2 it can be seen that ginsenoside/insulin prepared by the present invention is in gel.
3 in-vitro percutaneous absorption experiment of embodiment：
1）The outer percutaneous absorbtion experiment of Mice Body
At room temperature, 50 volume BCA reagent As are added into 1 volume BCA reagents B（50:1）Appropriate BCA working solutions are prepared, are mixed well.
BCA working solutions room temperature is stablized in 24 hours.It is completely dissolved protein standard substance, takes 10 μ L to be diluted to 100 μ L, keeps its final concentration of
Standard items (0.5mg/ml) after dilution are added to by 0,2,4,6,8,12,16,20 μ L Fen Do in EP pipes, are added by 0.5mg/ml
PBS is supplied to 20 μ L.200 μ L BCA working solutions are added in each pipe, 37 DEG C are placed 30min, finally the wavelength between 562nm
Light absorption value, draw the standard curve of protein quantification, the results are shown in Figure 3.
As seen from Figure 3, it is linearly good to measure protein content for BCA methods.
2）Extracorporeal releasing test
At room temperature, the ginsenoside of prepared 1.6mg/mL-insulin nano gel is placed in the bag filter of 10000Da,
Bag filter is placed in the PBS buffer solution of 30mL, pH7.4 again, sample is placed on 37 DEG C, the constant-temperature table of rotating speed 220r/min
On, it is measured respectively at different point in time sampling 1mL, and fill into 1mL PBS plain buffers simultaneously.According to BCA albumen
Quantitation curves calculate the cumulative release amount of insulin in nanogel, and the results are shown in Figure 4.
From fig. 4, it can be seen that its external 48h releasably about 80%.
3）The outer percutaneous absorbtion experiment of Mice Body
6 complete fresh mouse part skins are prepared, in-vitro percutaneous absorption is carried out using improved Franz vertical types diffusion cell
Experiment.15mLPBS is added in reception tank.Skin is placed in position among supply pool and reception tank, by ginseng soap after fixing
Glycosides/insulin gelling agent is put into supply pool, proceeds by experiment.The effective infiltrating area of diffusion cell is 2.80 cm2, reception tank
Volume is 15mL.Diffusion cell being placed in 37 DEG C ± 0.5 DEG C of constant temperature water bath, magnetic stirring apparatus is placed under constant temperature water bath,
Mixing speed is 600 rmin/1.Receiving liquid is quantified respectively at 12,24,36,48,60,72,84,96,120,144,168h
It takes out, while mending with the blank receiving liquid of same volume, record takes out the volume of acceptable solution.It is bent according to BCA albuminimetries standard
The accumulation transdermal release amount of nanogel prepared by the line computation present invention, the results are shown in Figure 5.
As seen from Figure 5, nanogel patch shows good slow controlled-release effect, and percutaneous rate is very fast in 0-12h, later
Reach one and relatively smoothly penetrates state.
Treatment of the 4 insulin nano gel preparation of embodiment to diabetic mice：
1）Modeling and grouping：The kunming mice of weight 18-20g, after normally raising 7 days, tail vein injection streptozotocin
（streptozotocin STZ）50~150mg/kg, if continuous two days blood glucose values are more than 11.1mmol/L or glucose
Level then thinks diabetic mice modeling success before blood glucose value when tolerance test 120min does not recover to injection still.By 30 at
The diabetic mice of work(modeling is randomly divided into nanogel group, injection of insulin group and control group.
Nanogel preparation cutaneous penetration：Mouse weight is weighed, carrying out gel nanometer formulation by 40mg/kg dosage percutaneously gives
Medicine is treated, and the hair of experiment mice abdomen is handled with electric shaver, mouse is fixed on plank after processing, is close to
2cm × 2cm percutaneous dosing control-released plasters are put back in mouse cage after the administration regular hour and are normally raised, and in different time points
Detect change of blood sugar.Injection of insulin：Abdomen injection administration is carried out by equivalent dose, i.e., is wiped experiment mice abdomen alcohol ball
Disinfection is wiped, is injected by the dosage of 40mg/kg, identical time point surveys blood glucose.Control group is without any processing.
Adverse reaction is observed：Diabetic mice is observed continuously 3 days after the treatment of nanogel percutaneous dosing and finds nothing
Apparent state of mind abnormality, each appetite and drinking-water are normal, do not find that red and swollen and hair occurs in the skin of therapentic part
Scorching phenomenon illustrates that transdermal skin patches medication is safe.
2）3 groups of mouse are administered simultaneously respectively, and point takes blood in different times, and the change of 3 groups of mouse blood sugars is surveyed with GLU kits
Change value, the results are shown in Figure 6.
As seen from Figure 6, injection of insulin group declines most fast, but then nanogel group mouse blood sugar is noted less than insulin
Group and control group are penetrated, and nanogel agent group blood glucose value rises to the rear trend that reduction is presented again for 24 hours, illustrates that the present invention prepares
Ginsenoside-insulin nano gel transdermal delivery system has the function of sustained release.
Claims (8)
1. a kind of ginsenoside-insulin nano gel, it is characterised in that：Certainly by ginsenoside gel nano particles and insulin
Assembling forms ginsenoside/insulin gel nano particles, then itself and matrix, moisturizer, penetration-assisting agent are mixed to prepare the people
Join saponin(e-insulin nano gel.
2. ginsenoside-insulin nano gel according to claim 1, it is characterised in that：The ginsenoside gel is received
The preparation of rice grain includes the following steps：
1）Using general ginsenoside as raw material, progress acidolysis reaction obtains ginsenoside acid hydrolysis solution in acidic aqueous solution；
2）By step 1）After the purified removal impurity of gained ginsenoside acid hydrolysis solution, it is re-dissolved in water to obtain the final product.
3. ginsenoside-insulin nano gel according to claim 2, it is characterised in that：Step 1）Described in ginseng it is total
Saponin(e is one or more in Araliaceae China ginseng, Korean ginseng, American Ginseng, Vietnam's ginseng, Japanese ginseng；Institute
State the aqueous solution that acidic aqueous solution is organic and or inorganic acids, pH value 1-3；The temperature of the acidolysis reaction be 50 ~
105 DEG C, the time is 1 ~ 72h.
4. ginsenoside-insulin nano gel according to claim 2, it is characterised in that：Step 2）Concrete operations be：
Ginsenoside acid hydrolysis solution is stood 2 hours in -25 DEG C~35 DEG C or more, then through centrifuging or filtering removal precipitation, clear liquid is used
Organic base and/or inorganic base adjust pH to 7.5~14, obtain sediment；At 25 DEG C~85 DEG C, sediment is dissolved in organic
In solvent, solution is then cooled to -20 DEG C~10 DEG C, is dried after centrifuging or filtering removal precipitation；
The organic solvent is dimethyl sulfoxide (DMSO), methanol, ethyl alcohol, n-butanol, propyl alcohol, tetrahydrofuran, one kind in pyridine or more
Kind；
The drying is constant pressure and dry, dry or spray drying is concentrated under reduced pressure.
5. ginsenoside-insulin nano gel according to claim 1, it is characterised in that：Ginsenoside/the insulin
The average grain diameter of gel nano particles is 30 ~ 100nm.
6. ginsenoside-insulin nano gel according to claim 1, it is characterised in that：The matrix includes natural high
It is one or more in molecule, semi-synthetic macromolecule and synthesis macromolecule；The moisturizer include glycerine, trimethylglycine,
It is one or more in saualane, cyclopolysiloxane, stearic acid, cholesterol, lecithin；The penetration-assisting agent is propylene glycol, azone
It is one or more in class, pyrrolones, fitter acids and its ester class, cyclodextrin, sulfoxide type.
7. a kind of preparation method of ginsenoside as described in claim 1-insulin nano gel, it is characterised in that：Including with
Lower step：
1）Insulin and ginsenoside gel nano particles are taken, secondary water is added, makes ginsenoside gel nano particles in solution
A concentration of 1 ~ 40 mg/mL, insulin a concentration of 1 ~ 25U/mL, ultrasonic vibration 15min is solidifying to get ginsenoside/insulin
Glue nanoparticles solution；
2）Matrix, moisturizer and penetration-assisting agent are dissolved in secondary water, ginsenoside/insulin gel is added under stirring conditions
Nanoparticles solution, up to ginsenoside-insulin nano gel after mixing well.
8. a kind of ginsenoside as described in claim 1-insulin nano gel be used to prepare with function of reducing blood sugar it is percutaneous to
Application in medicine preparation, it is characterised in that：The percutaneous drug administration preparation includes ointment, patch, spray.
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Cited By (1)
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| CN109875956A (en) * | 2019-04-03 | 2019-06-14 | 福州大学 | A kind of ginsenoside-warfarin self-assembled nanometer gel and its application |
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