CN104353062A - Insulin oral nano-preparation and preparation method thereof - Google Patents
Insulin oral nano-preparation and preparation method thereof Download PDFInfo
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- CN104353062A CN104353062A CN201410671032.4A CN201410671032A CN104353062A CN 104353062 A CN104353062 A CN 104353062A CN 201410671032 A CN201410671032 A CN 201410671032A CN 104353062 A CN104353062 A CN 104353062A
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Abstract
The invention discloses an insulin oral nano-preparation and a preparation method thereof. Polymer sulfhydrylated hyaluronic acid with biocompatibility and bio-adhesion is adopted as a carrier to prepare an insulin-entrapped nano-suspension, and a lyophilized preparation is finally prepared. According to the method, the bio-adhesive material sulfhydrylated hyaluronic acid is firstly used as the carrier to prepare sulfhydrylated hyaluronic acid nano-particles, the insulin oral bioavailability can be improved by the preparation, and the preparation has the characteristic of excellent stability, is easy to store and transport, is convenient for patient using, and is a preparation with wide development prospect.
Description
Technical field
The invention belongs to field of medicine preparing technology, be specifically related to a kind of oral insulin nanometer formulation and preparation method thereof.
Background technology
The number that the whole world suffers from diabetes reaches 3.5 hundred million, and the trend risen year by year in addition, wherein the patient groups of about 1/4 suffers from type 1 diabetes, needs to use insulin throughout one's life.At present, insulin dosage form conventional is clinically subcutaneous injection dosage form, there is the problems such as the inconvenient and untoward reaction of patient compliance difference, long-term prescription is many, therefore, develops a kind of oral insulin dosage form and have important clinical meaning.
But insulin is as a kind of albumen, polypeptide drug, and it is large that its oral administration also exists molecular weight, and water solublity is strong, and film permeability is poor, and biological half-life is short, easily by obstacles such as gastrointestinal tract environment destructions, makes its oral administration biaavailability be only 0.1%-2%.For overcoming above malabsorption, adopt biological adhesion type nano-delivery system as drug delivery system, both can resist after combining nano carrier bag medicine carrying thing gastrointestinal tract environment destroy and particle diameter little, be easy to be improved its bioavailability by effects such as intestinal M cytophagies, again can by the holdup time of bioadhesive carrier prolong drug on intestinal mucosa, improve the local concentration of slime layer medicine, make to stick position and produce the driving force that promotes cell Passive intake medicine, thus improve the oral absorption of medicine.
Summary of the invention
The object of the present invention is to provide a kind of oral insulin nanometer formulation and preparation method thereof, first biological adhesive material sulfhydryl hyaluronic acid is prepared thiolated hyaluronic acid nanoparticle as carrier, said preparation can improve oral insulin bioavailability, and have and have good stability, being easy to store, transport and be convenient to the features such as patient's use, is a kind of preparation with wide DEVELOPMENT PROSPECT.
For achieving the above object, the present invention adopts following technical scheme:
A kind of oral insulin nanometer formulation take thiolated hyaluronic acid as carrier material, and preparation bag carries the nanosuspension of insulin, obtains through lyophilization.
The molecular weight ranges of described thiolated hyaluronic acid is 400kDa-2000kDa, and free sulfhydryl groups content is 50-400 μm of olg
-1.
A kind of method preparing oral insulin nanometer formulation as above: first using thiolated hyaluronic acid, insulin and surfactant dissolves in tri-distilled water as aqueous phase, again aqueous phase is joined in organic facies, Probe Ultrasonic Searching or high pressure homogenize obtain colostrum, aqueous medium is added after reduction vaporization removing organic solvent, Probe Ultrasonic Searching or high pressure homogenize again, obtain nanosuspension, obtain oral insulin nanometer formulation through lyophilization.
In aqueous phase, the mass ratio of thiolated hyaluronic acid and insulin is 0.5:1-10:1.
Surfactant is one or more in Tween 80, soybean lecithin, Ovum Gallus domesticus Flavus lecithin, oleic acid polyethyleneglycol glyceride, sad certain herbaceous plants with big flowers acid polyethylene glycol glyceride, Polyethylene Glycol 40 castor oil hydrogenated, CREMOPHORE EL, HS15, and consumption is 0.1%-10%(w/v).
Organic facies is one or more in dichloromethane, acetone, cyclohexane extraction, normal hexane, ether; Containing surfactant in organic facies, described surfactant is one or more in oleic acid polyethyleneglycol glyceride, sorbester p17, sad certain herbaceous plants with big flowers acid polyethylene glycol glyceride, Polyethylene Glycol 40 castor oil hydrogenated, CREMOPHORE EL, HS15, and content is 0.1%-10%(w/v).
The volume ratio of aqueous phase and organic facies is 5:1-1:20.
Probe Ultrasonic Searching technological parameter: power is 100W-500W, ultrasonic time is 30 seconds-10 minutes, and temperature is 0 DEG C-40 DEG C.
High pressure homogenize technological parameter: pressure is 800-1200bar, circulation 3-10 time.
During lyophilization, need to add freeze drying protectant in nanosuspension, described freeze drying protectant is one or more in glucose, lactose, mannitol, trehalose, and consumption is 2%-20%(w/w).
Remarkable advantage of the present invention is:
(1) the present invention adopts the polymer thiolated hyaluronic acid with good biocompatibility and bioadhesive as pharmaceutical carrier, prepare the thiolated hyaluronic acid nanoparticle that bag carries insulin, thus improve medicine in the gastrointestinal holdup time, improve its oral administration biaavailability.Biological adhesive material sulfhydryl hyaluronic acid is carried the carrier material of insulin by the present invention first as bag, biocompatibility is good, is a kind of novel insulin oral Preparation, can be applied to clinical, have wide DEVELOPMENT PROSPECT.
(2) obtained insulin nano suspension has been prepared into final lyophilized formulations by cryodesiccated method by the present invention, extends the effect duration of preparation, is conducive to the storage of medicine, transport, facilitates patient and uses, thus have good DEVELOPMENT PROSPECT.
Accompanying drawing explanation
Fig. 1 is insulin nano preparation transmission electron microscope picture.
Fig. 2 is the potential change figure before and after oral insulin nanoparticle mixes with mucoprotein microgranule.
Fig. 3 is blood sugar level variation diagram after diabetes rat administration.
Detailed description of the invention
There is the preparation method of the oral insulin nanometer formulation of bio-adhesive performance: first adopt the standby bag of second emulsifying legal system to carry the thiolated hyaluronic acid nanoparticle suspension of insulin, then adopt Freeze Drying Technique to be prepared into lyophilized formulations.
(1) preparation of nanosuspension: take thiolated hyaluronic acid, insulin is dissolved in aqueous phase, wherein the mass ratio of thiolated hyaluronic acid and insulin is 0.5:1-10:1; Add surfactant in aqueous phase and can be one or more mixture in Tween 80, soybean lecithin, Ovum Gallus domesticus Flavus lecithin, oleic acid polyethyleneglycol glyceride, sad certain herbaceous plants with big flowers acid polyethylene glycol glyceride, Polyethylene Glycol 40 castor oil hydrogenated, CREMOPHORE EL, HS15, consumption is 0.1%-10%(w/v); Organic facies is one or more mixture in dichloromethane, acetone, cyclohexane extraction, normal hexane, ether; Adding surfactant in organic facies is one or more mixture in oleic acid polyethyleneglycol glyceride, sorbester p17, sad certain herbaceous plants with big flowers acid polyethylene glycol glyceride, Polyethylene Glycol 40 castor oil hydrogenated, CREMOPHORE EL, HS15, and consumption is 0.1%-10%(w/v); Aqueous phase and organic facies volume ratio are 5:1-1:20; Probe Ultrasonic Searching power is 100W-500W, and ultrasonic time is 30 seconds-10 minutes, and temperature is 0 DEG C-40 DEG C, or adds in high pressure homogenizer, and pressure is 800-1200bar, and circulation 3-10 time, obtains colostrum; Reduction vaporization removing organic solvent, then add aqueous phase, Probe Ultrasonic Searching or high pressure homogenize, obtain nanosuspension again.
(2) preparation method of final lyophilized formulations, is characterized in that, obtained nanosuspension adds one or more that freeze drying protectant can be in glucose, lactose, mannitol, trehalose, and consumption is 2%-20%(w/w).
The preparation of embodiment 1 oral insulin nanometer formulation
Preparation technology: get thiolated hyaluronic acid and insulin is dissolved in distilled water, add Tween 80, mix homogeneously is as aqueous phase, aqueous phase is slowly added under magnetic stirring in the acetone containing CREMOPHORE EL, Probe Ultrasonic Searching 300W, 5min, obtain w/o type colostrum; 40 DEG C of rotary evaporation removing acetone, adding distil water is to 200ml, and Probe Ultrasonic Searching 300W, 5min again, obtains insulin nanoparticles suspension; In insulin nanoparticles suspension, add lactose and mannitol as freeze drying protectant ,-70 DEG C of pre-freeze 8h, lyophilization 48h, to obtain final product.
The preparation of embodiment 2 oral insulin nanometer formulation
Preparation technology: get thiolated hyaluronic acid and insulin is dissolved in distilled water, add Tween 80, mix homogeneously is as aqueous phase, and slowly added under magnetic stirring by aqueous phase in the dichloromethane containing soybean phospholipid, Probe Ultrasonic Searching 250W, 3min, obtain w/o type colostrum; 40 DEG C of rotary evaporation removing acetone, adding distil water is to 200ml, and Probe Ultrasonic Searching 250W, 3min again, obtains insulin nanoparticles suspension; In insulin nanoparticles suspension, add glucose and mannitol as freeze drying protectant ,-70 DEG C of pre-freeze 8h, lyophilization 48h, to obtain final product.
pharmacodynamic experiment result in the rat body of oral insulin nanometer formulation
Choose fasting glucose between 2.9 ~ 6.9 mmolL
-1male SD rat be experimental subject, fasting before SD rat experiment (can't help water) 12 h, after weighing, by 55 mgkg
-1dosage take streptozotocin (STZ), with citrate buffer solution (0.1 molL
-1, pH4.5) dissolve, lumbar injection is in rat body immediately.Stood upside down by rat immediately after having injected medicine, this measure is conducive to medicine and flows to the lower pancreatic sites in position at intraperitoneal, can contact failure islet cells direct with pancreas.72 h after modeling, survey blood glucose, and normal raising is after 1 week, and from tail vein blood, measure by Bai Anjie blood glucose meter, blood glucose value is more stable, higher than 16.7 mmolL
-1rat can be applicable to following experiment.Observe and survive quantity after rat modeling, amount of drinking water, voided volume and the general state such as to lose weight.
Successful for modeling SD rat 20 is divided into 4 groups at random, and the 1st group is positive controls, subcutaneous injection insulin-containing 5 Ukg
-1insulin solutions; 2nd group of gavage gives insulin-containing 50 Ukg
-1insulin solutions; 3rd group of gavage gives 50 Ukg
-1ins-HA-Cys-NPs; 4th group of gavage gives 50 Ukg
-1ins-HA-NPs.Respectively at 0,0.25,0.5,1,2,3,4,6,8,10 and 12 h, cut tail and get blood, survey blood glucose by Bai Anjie blood glucose meter.With zero time blood glucose value for 100%, calculate each time point blood glucose percentage rate.
Adopt the relative pharmacological bioavailability (relative pharmacological availability, PA) of following formulae discovery insulin:
In formula, AAC represent with trapezoidal method calculate 0 to 12 h in blood glucose curve on area " sc, oral " represent subcutaneous administrations and Oral administration respectively.
result
Experimental result shows, rat skin lower injection insulin solutions (5 Ukg
-1), relative blood sugar level comparatively administration front decline during 1 h after administration, difference has statistical significance (p<0.05), blood glucose value gos up after reaching minimum fast, during 6 h, blood glucose value reverts to original 76.6%, and after Oral Administration in Rats insulin nano preparation, show slower hypoglycemic activity, the rat of oral insulin nanometer formulation has significant difference (P<0.05) before the relative blood sugar level of 4 h ~ 12 h comparatively administration, blood glucose value is minimum at 4 h, and 8 h blood glucose values are original 72.6%.Compared with subcutaneous administrations mode, the rat relative pharmacological bioavailability of oral insulin nanometer formulation is 6.9%.Compared with direct oral insulin solution, there is significant hypoglycemic effect after showing oral insulin nanometer formulation,
table 1rat Internal pharmacokinetics parameter after different insulin preparation administration (
, n=5)
The foregoing is only preferred embodiment of the present invention, all equalizations done according to the present patent application the scope of the claims change and modify, and all should belong to covering scope of the present invention.
Claims (10)
1. an oral insulin nanometer formulation, is characterized in that: take thiolated hyaluronic acid as carrier material, and preparation bag carries the nanosuspension of insulin, obtains oral insulin nanometer formulation through lyophilization.
2. oral insulin nanometer formulation according to claim 1, is characterized in that: the molecular weight ranges of described thiolated hyaluronic acid is 400kDa-2000kDa, and free sulfhydryl groups content is 50-400 μm of olg
-1.
3. prepare the method for oral insulin nanometer formulation as claimed in claim 1 for one kind, it is characterized in that: first using thiolated hyaluronic acid, insulin and surfactant dissolves in tri-distilled water as aqueous phase, again aqueous phase is joined in organic facies, Probe Ultrasonic Searching or high pressure homogenize obtain colostrum, aqueous medium is added after reduction vaporization removing organic solvent, Probe Ultrasonic Searching or high pressure homogenize again, obtains nanosuspension, obtains oral insulin nanometer formulation through lyophilization.
4. method according to claim 3, is characterized in that: in aqueous phase, the mass ratio of thiolated hyaluronic acid and insulin is 0.5:1-10:1.
5. method according to claim 3, it is characterized in that: surfactant is one or more in Tween 80, soybean lecithin, Ovum Gallus domesticus Flavus lecithin, oleic acid polyethyleneglycol glyceride, sad certain herbaceous plants with big flowers acid polyethylene glycol glyceride, Polyethylene Glycol 40 castor oil hydrogenated, CREMOPHORE EL, HS15, and consumption is 0.1%-10%(w/v).
6. method according to claim 3, is characterized in that: organic facies is one or more in dichloromethane, acetone, cyclohexane extraction, normal hexane, ether; Containing surfactant in organic facies, described surfactant is one or more in oleic acid polyethyleneglycol glyceride, sorbester p17, sad certain herbaceous plants with big flowers acid polyethylene glycol glyceride, Polyethylene Glycol 40 castor oil hydrogenated, CREMOPHORE EL, HS15, and content is 0.1%-10%(w/v).
7. method according to claim 3, is characterized in that: the volume ratio of aqueous phase and organic facies is 5:1-1:20.
8. method according to claim 3, is characterized in that: Probe Ultrasonic Searching technological parameter: power is 100W-500W, and ultrasonic time is 30 seconds-10 minutes, and temperature is 0 DEG C-40 DEG C.
9. method according to claim 3, is characterized in that: high pressure homogenize technological parameter: pressure is 800-1200bar, circulation 3-10 time.
10. method according to claim 3; it is characterized in that: during lyophilization; need in nanosuspension to add freeze drying protectant, described freeze drying protectant is one or more in glucose, lactose, mannitol, trehalose, and consumption is 2%-20%(w/w).
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| CN106334185A (en) * | 2016-08-25 | 2017-01-18 | 广东省人民医院 | Polypeptide drug-containing self-nano-emulsion oral preparation and preparation method thereof |
| CN107530296A (en) * | 2015-04-21 | 2018-01-02 | 北卡罗来纳州立大学 | Use the glucose responding insulin delivery system of hypoxia sensitivity nano composite material |
| CN108371708A (en) * | 2018-02-02 | 2018-08-07 | 中山大学 | A kind of oral insulin nanoparticle formulations and preparation method thereof |
| CN108653233A (en) * | 2017-04-01 | 2018-10-16 | 苏州苏融生物医药有限公司 | It is a kind of that there is bioadhesive to carry polypeptide protein class medical solid particulate matter, the preparation comprising it, Preparation method and use |
| WO2019076125A1 (en) * | 2017-10-20 | 2019-04-25 | 中山大学 | Nanoparticle loaded with therapeutic protein and microcapsule thereof |
| CN111417387A (en) * | 2017-07-12 | 2020-07-14 | 罗盖特兄弟公司 | Cross-linked maltodextrin for oral delivery of biologically active substances |
| US11351230B2 (en) | 2016-11-07 | 2022-06-07 | North Carolina State University | Patch loaded with dual-sensitive vesicles for enhanced glucose-responsive insulin delivery |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107530296A (en) * | 2015-04-21 | 2018-01-02 | 北卡罗来纳州立大学 | Use the glucose responding insulin delivery system of hypoxia sensitivity nano composite material |
| CN113350489A (en) * | 2015-04-21 | 2021-09-07 | 北卡罗来纳州立大学 | Glucose responsive insulin delivery system using hypoxia sensitive nanocomposite material |
| US11191815B2 (en) | 2015-04-21 | 2021-12-07 | North Carolina State University | Glucose-responsive insulin delivery microneedle system |
| CN106334185A (en) * | 2016-08-25 | 2017-01-18 | 广东省人民医院 | Polypeptide drug-containing self-nano-emulsion oral preparation and preparation method thereof |
| CN106334185B (en) * | 2016-08-25 | 2020-01-10 | 广东省人民医院 | Oral preparation containing polypeptide drug self-nanoemulsion and preparation method thereof |
| US11351230B2 (en) | 2016-11-07 | 2022-06-07 | North Carolina State University | Patch loaded with dual-sensitive vesicles for enhanced glucose-responsive insulin delivery |
| CN108653233A (en) * | 2017-04-01 | 2018-10-16 | 苏州苏融生物医药有限公司 | It is a kind of that there is bioadhesive to carry polypeptide protein class medical solid particulate matter, the preparation comprising it, Preparation method and use |
| CN111417387A (en) * | 2017-07-12 | 2020-07-14 | 罗盖特兄弟公司 | Cross-linked maltodextrin for oral delivery of biologically active substances |
| CN111417387B (en) * | 2017-07-12 | 2022-12-23 | 罗盖特兄弟公司 | Cross-linked maltodextrin for oral delivery of biologically active substances |
| WO2019076125A1 (en) * | 2017-10-20 | 2019-04-25 | 中山大学 | Nanoparticle loaded with therapeutic protein and microcapsule thereof |
| CN108371708A (en) * | 2018-02-02 | 2018-08-07 | 中山大学 | A kind of oral insulin nanoparticle formulations and preparation method thereof |
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Application publication date: 20150218 |