CN103142475B - Exenatideacetate sustained-release microsphere preparation and preparation method thereof - Google Patents
Exenatideacetate sustained-release microsphere preparation and preparation method thereof Download PDFInfo
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- CN103142475B CN103142475B CN201210494756.7A CN201210494756A CN103142475B CN 103142475 B CN103142475 B CN 103142475B CN 201210494756 A CN201210494756 A CN 201210494756A CN 103142475 B CN103142475 B CN 103142475B
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Abstract
The invention discloses an exenatideacetate acetate sustained-release microsphere preparation and a preparation method thereof. The sustained-release microsphere comprises, calculated by the weight of the microsphere, 0.1%-20% (w / W) of exenatideacetate, 80%-99.9% of a biodegradable and biocompatible polymer material with molecular weight of 10000-100000 Dalton, and 0%-10% of a pharmaceutically acceptable emulsion and an excipient. The sustained-release microsphere provided by the invention has average particle size of 5-50 mum and encapsulation efficiency greater than 92%. The sustained-release microsphere has a release period of up to more than 40 days, and can significantly reduce the frequency of medication, improve bioavailability of nesiritide acetate, reduce side effect of the drug, and facilitate clinical treatment.
Description
Technical field:
The present invention relates to a kind of sustained release microsphere agents for the treatment of diabetes, be specifically related to a kind of containing Exendin-4 sustained release microsphere agents and preparation method.
Background technology:
Exenatide is to separate from the salivation thing of Monster at first, is a kind of 39 amino acid whose polypeptide that contain, and its aminoacid mechanism is as follows:
His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met-Glu-Glu-Glu-Ala-Val-Arg-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH
2。
The aminoacid sequence of Exenatide and mankind GLP-1 partly overlap.Exenatide show in vitro can in conjunction with and activate known mankind GLP-1 receptor, this just mean by comprise cAMP and/or other Cellular Signaling Transduction Mediated mechanism make dependence on the glucose insulin synthesis and beta Cell of islet in vivo excreting insulin increase.In the situation that concentration of glucose raises, Exenatide can promote insulin to discharge from β cell.Certain antihyperglycemic effect of Exenatide simulation GLP-1 after vivo medicine-feeding.
Summary of the invention:
The technical problem to be solved in the present invention is to provide a kind of can effectively extend Exendin-4 Exendin-4 sustained release microsphere agents of action time in vivo, said preparation can reduce frequency injection and Drug tolerance, improve patient's adaptability, facilitate clinical use and patient to accept.In addition, it has also eliminated the vivo medicine concentration peak valley phenomenon that normal injection agent multiple dosing produces, and can obtain steadily valid density for a long time, has reduced toxic and side effects, and total dosage reduces.
In addition, the present invention also provides a kind of preparation method of Exendin-4 sustained release microsphere agents.
The present invention has prepared a kind of sustained release microsphere agents of the injection containing Exendin-4 medicine, it is characterized in that, acetic acid Exendin-4 sustained-release micro-spheres in described preparation is containing the Exendin-4 that accounts for microsphere weight 0.1%-20% (w/w), the molecular weight that accounts for microsphere weight 80%-99.9% is 5,000-300, the macromolecular material of 000 daltonian biodegradable and tool biocompatibility, and pharmaceutically acceptable other adjuvants that account for microsphere weight 0%-10%.Described sustained-release micro-spheres particle diameter is 1-200 μ m, average 5-100 μ m.
The macromolecular material of described biodegradable and tool biocompatibility, optional from polylactide (PLA), PGA (PGA), polylactide-co-glycolide (PLGA), polyalkylcyanoacrylate, polycaprolactone (PCL), poly hydroxybutyric acid (PHB), poly-hydroxyl valeric acid (PVA), poly-capric acid (PDA), poly-anhydride, Polyhydroxybutyrate-co-hydroxyvalerate, polylactic acid-polyglycol, polyglycolic acid-Polyethylene Glycol a kind of or its mixture wherein, preferably PLGA.Described lactide and Acetic acid, hydroxy-, bimol. cyclic ester molecular weight ranges be all at 5,000-20,000 dalton, and the proportion of lactide and Acetic acid, hydroxy-, bimol. cyclic ester is 15: 85-85: 15.
Described pharmaceutically acceptable other adjuvants comprise emulsion stabilizer and excipient, and described emulsion stabilizer is selected from polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), sodium polymethacrylate, sodium polyacrylate, sodium carboxymethyl cellulose etc.Emulsion stabilizer preferably polyethylene alcohol, its amount ranges is 0.1%-5%.
Excipient is selected from a kind of or its mixture in sorbitol, mannitol, lactose, sucrose, glycine, the mixture of preferred mannitol and lactose, and its amount ranges is 0.1%-5%.
In addition, the present invention also provides the method first step of preparation containing acetic acid Exendin-4 medicine, first that Exendin-4 is water-soluble, obtains interior water; Separately PLGA is dissolved in organic solvent, obtains oil phase; Oil phase and interior water are placed in agitator, and breast is even at a high speed, forms W/O emulsion; Then W/O emulsion is joined in polyvinyl alcohol water solution, breast is even at a high speed, forms W/O/W emulsion;
Second step, moves into W/O/W emulsion in polyvinyl alcohol water solution, and stirring at low speed is centrifugal, collects thus obtained microsphere, with distilled water wash repeatedly after, more centrifugal collection, lyophilization, obtains Exendin-4 sustained-release micro-spheres.
accompanying drawing explanation:
Take drug release time as abscissa, and accumulative releasing degree is vertical coordinate, draws the Exendin-4 sustained release microsphere agents tablets in vitro curve chart of embodiment 1,2,3 preparations, the results are shown in Figure of description 1,2,3.
Accompanying drawing 1 is embodiment 1 release in vitro curve chart;
Accompanying drawing 2 is embodiment 2 release in vitro curve charts;
Accompanying drawing 3 is embodiment 3 release in vitro curve charts.
the specific embodiment:
Below in conjunction with embodiment, embodiment of the present invention are described in detail, but it will be understood to those of skill in the art that the following example is only for the present invention is described, and should not be considered as limiting scope of the present invention.
Embodiment 1
Take 10mg Exendin-4 and be dissolved in distilled water, obtain interior water; Take 1000mgPLGA (polymerization ratio=20: 80) be dissolved in dichloromethane, obtain oil phase.Compound concentration is the PVA solution 500ml that 15% PVA solution 50ml and concentration are 1.5%.First Exendin-4 solution is moved into and is dissolved with in the dichloromethane solution of PLGA, under room temperature, be placed on emulsion dispersion machine at a high speed (30000rpm) breast even 30 seconds, then the w/o type Emulsion of gained is transferred to 50ml concentration and is in 15% PVA solution, be placed on emulsion dispersion machine the rotating speed with 5000rpm, even 1 minute of breast, obtain W/O/W type emulsion, move in the PVA solution of 500ml1.5, be placed on mechanical agitator, with the rotating speed stirring at low speed of 500rpm 2 hours, centrifugal, collect thus obtained microsphere, with distilled water, repeatedly wash, and then centrifugal collection, lyophilization, being distributed into actual drug loading is the Exendin-4 sustained release microsphere agents of 5.0mg dosage, thus obtained microsphere particle diameter is 10-50 μ m, mean diameter is 30 μ m.
The medicine carrying Exendin-4 sustained-release micro-spheres 5mg that precision takes embodiment 1 preparation is dissolved in 1mL dichloromethane, gradation adds 3mL and 2mL double distilled water to extract medicine, vortex extraction 1min, the centrifugal 3min of 15000rpm, draw and merge supernatant, pipette supernatant 0.5mL and be settled to 10mL with double distilled water, sample introduction 20 μ l, HPLC measures content, drawing standard curve, the medicine carrying Exendin-4 sustained-release micro-spheres Chinese medicine content that calculates embodiment 1 preparation according to standard curve, calculated yield and envelop rate drug loading are respectively 91.5% and 84.8% accordingly.
Embodiment 2
Take 15mg Exendin-4 and be dissolved in distilled water, obtain interior water; Take 1000mgPLGA (polymerization ratio=40: 60) be dissolved in dichloromethane, obtain oil phase.Compound concentration is the PVA solution 500ml that 10% PVA solution 50ml and concentration are 1%.First Exendin-4 solution is moved into and is dissolved with in the dichloromethane solution of PLGA, under room temperature, be placed on emulsion dispersion machine at a high speed (30000rpm) breast even 30 seconds, then the w/o type Emulsion of gained is transferred to 50ml concentration and is in 10% PVA solution, be placed on emulsion dispersion machine the rotating speed with 5000rpm, even 1 minute of breast, obtain W/O/W type emulsion, move in the PVA solution of 500ml1%, be placed on mechanical agitator, with the rotating speed stirring at low speed of 500rpm 2 hours, centrifugal, collect thus obtained microsphere, with distilled water, repeatedly wash, and then centrifugal collection, lyophilization, be distributed into actual drug loading at the Exendin-4 sustained release microsphere agents of 4.0mg dosage, thus obtained microsphere particle diameter is 5-40 μ m, mean diameter is 20 μ m.
The medicine carrying Exendin-4 sustained-release micro-spheres 5mg that precision takes embodiment 2 preparations is dissolved in 1mL dichloromethane, gradation adds 3mL and 2mL double distilled water to extract medicine, vortex extraction 1min, the centrifugal 3min of 15000rpm, draw and merge supernatant, pipette supernatant 0.5mL and be settled to 10mL with double distilled water, sample introduction 20 μ l, HPLC measures content, drawing standard curve, the medicine carrying Exendin-4 sustained-release micro-spheres Chinese medicine content that calculates embodiment 2 preparations according to standard curve, calculated yield and envelop rate drug loading are respectively 81.4% and 75.9% accordingly.
Embodiment 3
Take 20mg Exendin-4 and be dissolved in distilled water, obtain interior water; Take 500mgPLA and 1500mgPLGA (polymerization ratio=60: 40) be dissolved in dichloromethane, obtain oil phase.Compound concentration is the PVA solution 1000ml that 5% PVA solution 100ml and concentration are 0.5%.First Exendin-4 solution is moved into and is dissolved with in the dichloromethane solution of PLA and PLGA, under room temperature, be placed on emulsion dispersion machine at a high speed (30000rpm) breast even 30 seconds, then the w/o type Emulsion of gained is transferred to 100ml concentration and is in 5% PVA solution, be placed on emulsion dispersion machine the rotating speed with 5000rpm, even 1 minute of breast, obtain W/O/W type emulsion, move in the PVA solution of 1000m10.5%, be placed on mechanical agitator, with the rotating speed stirring at low speed of 500rpm 2 hours, centrifugal, collect thus obtained microsphere, with distilled water, repeatedly wash, and then centrifugal collection, lyophilization, be distributed into actual drug loading at the Exendin-4 sustained release microsphere agents of 3.5mg dosage, thus obtained microsphere particle diameter is 10-30 μ m, mean diameter is 25 μ m.
The medicine carrying Exendin-4 sustained-release micro-spheres 5mg that precision takes embodiment 3 preparations is dissolved in 1mL dichloromethane, gradation adds 3mL and 2mL double distilled water to extract medicine, vortex extraction 1min, the centrifugal 3min of 15000rpm, draw and merge supernatant, pipette supernatant 0.5mL and be settled to 10mL with double distilled water, sample introduction 20 μ l, HPLC measures content, drawing standard curve, the medicine carrying Exendin-4 sustained-release micro-spheres Chinese medicine content that calculates embodiment 3 preparations according to standard curve, calculated yield and envelop rate drug loading are respectively 57.2% and 60.7% accordingly.
Embodiment 4
To the Exendin-4 sustained release microsphere agents of embodiment 1,2,3 preparations is carried out to tablets in vitro investigation, method is as follows respectively:
The accurate Exendin-4 sustained-release micro-spheres that takes some parts of embodiment 1,2,3 preparations respectively, every part of 5mg, is placed in the cillin bottle of 35 10mL, and every part adds the Na that contains 0.1M
2hPO
4naH with 0.1M
2pO
4the phosphate buffer solution of pH7.4, be placed in 37 ℃ of waters bath with thermostatic control, respectively the 0th, 4, 8, 12, 16, 20, 24, 28, within 32 days, take out, centrifugal, again be dispersed in acetate dichloromethane buffer (1: 1v/v), chromatographic column is C1850 * 2mm, mobile phase is 1: 1 containing 0.1% trifluoracetic acid and containing 1% trifluoroacetic 50% acetonitrile mixed solution, flow velocity 1.0ml/min, at 240nm place, detect, measure remaining dose in microsphere, according to external standard method, calculate accumulative releasing degree, embodiment 1, 2, the tablets in vitro measurement result of the Exendin-4 sustained release microsphere agents of 3 preparations is as shown in table 1:
The tablets in vitro experimental result of the Exendin-4 sustained release microsphere agents of table 1 embodiment 1,2,3 preparations
Take drug release time as abscissa, and accumulative releasing degree is vertical coordinate, draws the Exendin-4 sustained release microsphere agents tablets in vitro curve of embodiment 1,2,3 preparations, the results are shown in Figure of description 1,2,3.
By yield and envelop rate, and release in vitro curve chart, the Exendin-4 sustained release microsphere agents of embodiment 1 preparation is better than embodiment 2, and embodiment 2 is better than embodiment 3.
Claims (3)
1. contain a sustained release microsphere agents for the injection of Exendin-4 medicine, it is characterized in that: contain 10mg Exendin-4,1000mgPLGA, lactide and Acetic acid, hydroxy-, bimol. cyclic ester polymerization ratio are 20:80, the PVA solution that emulsifying agent is 15%, its preparation technology is as follows: 10mg Exendin-4 is dissolved in distilled water, obtains interior water, 1000mgPLGA, lactide and Acetic acid, hydroxy-, bimol. cyclic ester polymerization ratio are 20:80, be dissolved in dichloromethane, obtain oil phase, interior water is moved into oil phase, under room temperature, be placed on emulsion dispersion machine the rotating speed with 30000rpm, even 30 seconds of breast, then the w/o type Emulsion of gained is transferred to 50mL concentration and is in 15% PVA solution, be placed on emulsion dispersion machine the rotating speed with 5000rpm, even 1 minute of breast, obtain W/O/W type emulsion, move in the PVA solution of 500mL1.5%, be placed on mechanical agitator, with the rotating speed stirring at low speed of 500rpm 2 hours, centrifugal, collect thus obtained microsphere, with distilled water, repeatedly wash, and then centrifugal collection, lyophilization, being distributed into actual drug loading is the Exendin-4 sustained release microsphere agents of 5.0mg dosage, thus obtained microsphere particle diameter is 10-50 μ m, mean diameter is 30 μ m.
2. contain a sustained release microsphere agents for the injection of Exendin-4 medicine, it is characterized in that: contain 15mg Exendin-4,1000mgPLGA, lactide and Acetic acid, hydroxy-, bimol. cyclic ester polymerization ratio are 40:60, the PVA solution that emulsifying agent is 10%, its preparation technology is as follows: 15mg Exendin-4 is dissolved in distilled water, obtains interior water, 1000mgPLGA, lactide and Acetic acid, hydroxy-, bimol. cyclic ester polymerization ratio are 40:60
,be dissolved in dichloromethane, obtain oil phase, interior water is moved into oil phase, under room temperature, be placed on emulsion dispersion machine the rotating speed with 30000rpm
,even 30 seconds of breast, then the w/o type Emulsion of gained is transferred to 50mL concentration and is in 10% PVA solution, be placed on emulsion dispersion machine the rotating speed with 5000rpm, even 1 minute of breast, obtain W/O/W type emulsion, move in the PVA solution of 500mL1%, be placed on mechanical agitator, with the rotating speed stirring at low speed of 500rpm 2 hours, centrifugal, collect thus obtained microsphere, with distilled water, repeatedly wash, and then centrifugal collection, lyophilization, being distributed into actual drug loading is the Exendin-4 sustained release microsphere agents of 4.0mg dosage, thus obtained microsphere particle diameter is 5-40 μ m, mean diameter is 20 μ m.
3. the sustained release microsphere agents containing the injection of Exendin-4 medicine, it is characterized in that: contain 20mg Exendin-4,500mgPLA and 1500mgPLGA, wherein PLGA, lactide and Acetic acid, hydroxy-, bimol. cyclic ester polymerization ratio are 60:40, the PVA solution that emulsifying agent is 5%, its preparation technology is as follows: 20mg Exendin-4 is dissolved in distilled water, obtains interior water, 500mgPLA and 1500mgPLGA, PLGA wherein, lactide and Acetic acid, hydroxy-, bimol. cyclic ester polymerization ratio are 60:40, be dissolved in dichloromethane, obtain oil phase, interior water is moved into oil phase, under room temperature, be placed on emulsion dispersion machine the rotating speed with 30000rpm, even 30 seconds of breast, then the w/o type Emulsion of gained is transferred to 100mL concentration and is in 5% PVA solution, be placed on emulsion dispersion machine the rotating speed with 5000rpm, even 1 minute of breast, obtain W/O/W type emulsion, move in the PVA solution of 1000mL0.5%, be placed on mechanical agitator, with the rotating speed stirring at low speed of 500rpm 2 hours, centrifugal, collect thus obtained microsphere, with distilled water, repeatedly wash, and then centrifugal collection, lyophilization, being distributed into actual drug loading is the Exendin-4 sustained release microsphere agents of 3.5mg dosage, thus obtained microsphere particle diameter is 10-30 μ m, mean diameter is 25 μ m.
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| CN104922660A (en) * | 2014-03-19 | 2015-09-23 | 陕西天森药物研究开发有限公司 | Application in treating diabetes and preparation method of exenatide long-acting sustain-released agent |
| WO2015158270A1 (en) * | 2014-04-16 | 2015-10-22 | 山东绿叶制药有限公司 | Exenatide-containing composition and preparation method therefor |
| CN107405307B (en) * | 2015-12-22 | 2020-11-10 | 四川科伦药物研究院有限公司 | Exenatide microsphere preparation and preparation method thereof |
| CN106729717A (en) * | 2016-12-12 | 2017-05-31 | 深圳市健翔生物制药有限公司 | The analogs of GLP 1 and ziconotide composition sustained-release microsphere preparation |
| CN108606957A (en) * | 2016-12-13 | 2018-10-02 | 南京星银药业集团有限公司 | A kind of oral slow-releasing preparation of the peptide containing Suo Malu and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101658496A (en) * | 2009-09-11 | 2010-03-03 | 中国人民解放军第二军医大学 | Exenatide release microsphere preparation, preparation method and application thereof |
| CN102198103A (en) * | 2011-05-30 | 2011-09-28 | 深圳翰宇药业股份有限公司 | Stable exenatide sustained-release microsphere preparation and preparation method thereof |
| CN102488619A (en) * | 2011-12-05 | 2012-06-13 | 上海交通大学 | Device for continuously producing exenatide microspheres and method for controlling release rate of microspheres |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101658496A (en) * | 2009-09-11 | 2010-03-03 | 中国人民解放军第二军医大学 | Exenatide release microsphere preparation, preparation method and application thereof |
| CN102198103A (en) * | 2011-05-30 | 2011-09-28 | 深圳翰宇药业股份有限公司 | Stable exenatide sustained-release microsphere preparation and preparation method thereof |
| CN102488619A (en) * | 2011-12-05 | 2012-06-13 | 上海交通大学 | Device for continuously producing exenatide microspheres and method for controlling release rate of microspheres |
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