CN1289066C - Glicetin -1 slow release microspheric preparation and its use - Google Patents
Glicetin -1 slow release microspheric preparation and its use Download PDFInfo
- Publication number
- CN1289066C CN1289066C CN 03151059 CN03151059A CN1289066C CN 1289066 C CN1289066 C CN 1289066C CN 03151059 CN03151059 CN 03151059 CN 03151059 A CN03151059 A CN 03151059A CN 1289066 C CN1289066 C CN 1289066C
- Authority
- CN
- China
- Prior art keywords
- medicine
- glp
- protective agent
- substrate
- oil phase
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Abstract
The present invention belongs to the technical field of medicine, which relates to a sustained-release microsphere preparation of glucagon like peptide-1(GLP-1) and a use thereof as hypoglycemic medicine or anti obesity medicine for controlling body weight. GLP-1 is endogenous polypeptide secreted by secretory cells of the small intestine and the large intestine and nerve cells of the brain and has the functions of stimulating insulin secretion, inhibiting glucagon secretion, inhibiting an appetite and delaying gastric emptying. The half-life of the GLP-1 in vivo is less than two minutes, and thus, the development and the utilization of the GLP-1 as medicine are restricted. In the present invention, the sustained-release microsphere preparation is prepared from the GLP-1 in the mode of PLGA as substrates, has the sustained-release effect of more than 4 weeks, and thus, is capable of being used as medicine for treating diabetes or controlling the body weight.
Description
Technical field
The present invention relates to medical technical field, is glucagon-like-peptide-1 (Glucagon-likepepide-1, sustained release microsphere agents GLP-1) and uses thereof.
Background technology
Glucagon-like-peptide-1 (Glucagon-like pepide-1, GLP-1) be by the secretory cell of small intestinal and large intestine and the excretory polypeptide of neurocyte of brain, be made of 37 aminoacid sequences, its aminoacid sequence is: His-Asp-Glu-Phe-Glu-Arg-His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Lle-Ala-Trp-Leu-Val-Lys-Gly-Arg-Gly.(Lopez?LC，Frazier?ML，Su?C-J，et?al.Mammalianpancreatic?proglucangon?contains?three?glucanguon-related?peptides.Proc?Nalt?Acad?Sci?USA?1983，80：5485-5489)。GLP-1 can stimulate secretion of insulin, and the secretion of energy glucagon suppression, reduce empty stomach or post-prandial glycemia, it acts on { the M.A.Nauck that has been confirmed in the clinical research of healthy human body and diabetic, D.Wollschlager, J.Werner, et al.Effects of subcutaneous glucagon-like peptide 1[GLP-1 (7-36amide)] in patients with NIDDM .Diabetologia1996,39:1546-1553}.Stimulate insulin secretion and the excretory effect of glucagon suppression just because of GLP-1 has, old friends attempt the medicine used as the treatment diabetes.Again can appetite-suppressing because of GLP-1, slow down gastric emptying, so also the someone plans to do it and is the medicine of controlling body weight (Juris J.Meier, Baptist Gallwitz, Wolfgang E.Schmidt, et al.Glucagon-like peptide-1as a regulator of food intake and body weight:therapeutic perspectves.Eur J Pharm 2000,440:269-279).GLP-1 is as endogenous polypeptide, safety and pharmacological action are clear and definite, toxic and side effects is very little, the hypoglycemic activity of GLP-1 is the concentration that depends on glucose in the blood plasma, be used for the treatment of diabetes and hypoglycemic side reaction can not take place, the diabetic that sulfonylurea hypoglycemic agent is failed to respond to any medical treatment has good curative effect equally.[Ritzel?R，Orskov?C，et?al.Pharmacokietic，insulinotropic，and?glucangonostatic?properties?ofGLP-1(7-36amide)after?subcutaneous?injection?in?healthy?volunteers.dose-response-relationships.Diabetologia?199538：720-725]。Yet, because GLP-1 half-life in vivo very short (less than 2 minutes) (DeaconCF, Knudsen LB, MadsenK, et al.Dipeptidyl peptidase IV resistant analogues of glucagons-likepeptide-1 which have extended metabolic stability and improvedbiological activity.Diabetologia 1998 41:271-278), this has just restricted its development and utilization as medicine, therefore is not applied to clinical so far yet.
Summary of the invention
The invention provides the sustained release microsphere agents of a kind of GLP-1, sustainable release reached more than 4 weeks, can be used for the treatment of diabetes or controlling body weight for the medicine of clinical use thereby make GLP-1 be expected to become.Biodegradable in recent years macromolecular material is widely used in the preparation of protein and peptide drugs slow-release microsphere, polylactic acid-glycolic guanidine-acetic acid copolymer [poly (lactide-co-glycolide) wherein, PLGA] because its excellent biological compatibility and biological degradability have been that pharmaceutical polymers uses by drugs approved by FDA.With PLGA as microsphere substrate, the inside and outside degradation experiment proves, in drug release process, whole polymer backbone is the uniformity degraded, decline along with molecular weight, the hydrophilic of framework material strengthens, because the continuous infiltration of moisture makes protein and peptide drugs continue to discharge (Wang YM, Sato H, Horikoshi.In vitro and in vivo evalution of taxol release from ploy (lactic-co-glycotic acid) microspheres containing isopropyl myristate anddegradation of the microspheres.J Control Rel, 1997,49:157).The present invention just is to use PLGA to prepare the GLP-1 sustained release microsphere agents as substrate.Because the polypeptide fragments that GLP-1 the 7th to the 36th amino acids is formed [being called for short GLP-1 (7-36)] has same biological activity with natural GLP-1, and more easily preparation, so the present invention is a principal agent with GLP-1 (7-36), PLGA is a substrate, and preparation is for hypodermic GLP-1 sustained release microsphere agents.
Sustained release microsphere agents preparation method of the present invention has three kinds:
1.w/o/w solvent evaporation method:
(1) preparation oil phase
Host material PLGA is dissolved in organic solvent dichloromethane makes oil phase, concentration is 50mg～300mg/ml;
(2) water in the preparation
Get water in the water-soluble formation of an amount of GLP-1 and protective agent, GLP-1 concentration is 1mg～10mg/ml, and protective agent is selected from zinc carbonate, human serum albumin, trehalose and mannitol etc., and its content is 1～5%;
(3) preparation microsphere
Interior water is added above-mentioned oil phase ultrasonic emulsification form colostrum, the rapid dropping of colostrum (also contained 0～2%NaCl in 2～6% polyvinyl alcohol (PVA) aqueous solution, or/and 0.5% surfactant F-68), (mixing speed is 1000～1800rpm) to the abundant homogenize of mechanical agitation, (mixing speed was 300～600rpm) in 4 hours to continue stirring at low speed under the room temperature, washing is collected, and lyophilization gets final product.
2.s/o/o solvent evaporation method:
(1) preparation oil phase
Host material PLGA is dissolved in organic solvent acetonitrile makes oil phase, concentration is 50mg～300mg/ml;
(2) preparation GLP-1 micropowder
An amount of Polyethylene Glycol (PEG) and GLP-1 and protective agent (its ratio is 8: 1: 2) are scattered in the water, after lyophilization, with washed with dichloromethane, centrifugal, remove PEG, obtain the 6LP-1 micropowder, protective agent is selected from zinc carbonate, trehalose and mannitol etc.;
(3) preparation microsphere
The GLP-1 micropowder is added oil phase, ultra-sonic dispersion, dropwise be added in the Oleum Gossypii semen then, (mixing speed is 600～800rpm) to the abundant homogenize of mechanical agitation, stirs 1 hour under the room temperature, and (mixing speed was 300～600rpm) in 2 hours to add an amount of petroleum ether continuation stirring again, can get the 6LP-1 microsphere, washing is collected, and lyophilization gets final product.
3. spray drying method.
(1) preparation GLP-1 micropowder
It is water-soluble to get an amount of GLP-1 and protective agent, and atomizing freeze drying obtains micropowder, and protective agent is selected from zinc carbonate, human serum albumin, trehalose etc., and its content is 1～5%;
(2) preparation microsphere
The GLP-1 micropowder is added organic solvent dichloromethane, ultra-sonic dispersion, spray drying promptly gets microsphere, and vacuum drying was collected after 6 hours under the room temperature.
The host material PL6A that the present invention is used, molecular weight are 3000～40000, polylactic acid (PLA): hydroxyacetic acid (PGA) is 25: 75～75: 25, and concentration is 50mg～300mg/ml.Ultrasonic emulsification, ultrasound condition is: duty cycle%:40; Output control:3 shelves; Timer:8 time.Inlet temperature is that 40 ℃, outlet temperature are 30 ℃, atomisation pressure 5Pa during spray drying.
Sustained-release micro-spheres of the present invention is through extracorporeal releasing experiment, and slow release reached more than 4 weeks, discharges to meet approximate zero mode, can be used for treating the subcutaneous injection preparation of diabetes and controlling body weight.
Description of drawings
Fig. 1 is the 1st day blood sugar concentration～time plot after the sustained release microsphere agents administration of GLP-1 of the present invention.
Fig. 2 is the 5th day blood sugar concentration～time plot after the sustained release microsphere agents administration of GLP-1 of the present invention.
Fig. 3 is the 10th day blood sugar concentration～time plot after the sustained release microsphere agents administration of GLP-1 of the present invention.
Fig. 4 is the 15th day blood sugar concentration～time plot after the sustained release microsphere agents administration of GLP-1 of the present invention.
Fig. 5 is the 20th day blood sugar concentration～time plot after the sustained release microsphere agents administration of GLP-1 of the present invention.
Fig. 6 is the 25th day blood sugar concentration～time plot after the sustained release microsphere agents administration of GLP-1 of the present invention.
Fig. 7 is the 30th day blood sugar concentration～time plot after the sustained release microsphere agents administration of GLP-1 of the present invention.
The specific embodiment
Embodiment 1:w/o/w solvent evaporation method prepares the GLP-1 sustained release microsphere agents
With PLGA (RG502H, PLA: PGA=50: 50, M
w=34000) 100mg is dissolved in the 1.0ml dichloromethane and makes oil phase, GLP-13mg is dissolved in the aqua bidestillata of 0.1ml and (includes 3% trehalose, 5% mannitol) form interior water, it is added above-mentioned oil phase, ultrasonic emulsification, form the colostrum of w/o, to contain 3%PVA solution 30ml (containing NaCl 2% and F-680.5%) and place stirred vessel, with colostrum abundant homogenize of aqueous phase outside high-speed stirred (1000rpm) adds down fast, after three minutes, rotating speed is adjusted downward to the simultaneously outer water of 400rpm adds the 30ml distilled water, stirred 4 hours microsphere sclerosis back centrifugalize and washing, lyophilization under the room temperature.Irradiation sterilization gets final product after the sealing packing.The envelop rate of GLP-l microsphere is 92%, particle diameter＜100 μ m.
Embodiment 2:s/o/o solvent evaporation method prepares the GLP-1 sustained release microsphere agents
PEG (PEG6000) 24mg and GLP-13mg and protective agent (zinc carbonate 5mg) are scattered in the lml aqua bidestillata, and vortex mixed about 3 minutes, after the lyophilization, with washed with dichloromethane, centrifugal, removed PEG, obtained the GLP-1 micropowder.With PLGA (RG502H, PLA: PGA=50: 50, M
w=34000) 200mg is dissolved in acetonitrile 1.0ml and makes oil phase, micropowder is added above-mentioned oil phase, and ultra-sonic dispersion dropwise adds fully homogenize in the Oleum Gossypii semen with it, stirred (600rpm) 1 hour, add an amount of petroleum ether again and continue to stir (400rpm) 2 hours, can get the GLP-1 microsphere, centrifugal, use petroleum ether, collect, lyophilization, irradiation sterilization gets final product after the sealing packing.The envelop rate of GLP-1 microsphere is 90%, particle diameter＜100 μ m.
Embodiment 3: spray drying method for preparation GLP-1 sustained release microsphere agents
GLP-18mg and protective agent (human serum albumin 30mg) are dissolved in the aqua bidestillata of 10ml, spray in the liquid nitrogen, liquid nitrogen is volatilized at low temperatures, obtain the GLP-1 micropowder.With PLGA (RG502H, PLA: PGA=50: 50, M
w=34000) 600mg is dissolved in dichloromethane 10ml and makes oil phase, and micropowder is added ultra-sonic dispersion in the oil phase, and spray drying, inlet temperature are that 40 ℃, outlet temperature are 30 ℃, atomisation pressure 5Pa, nozzle diameter 0.5mm, flow velocity 1～2ml/min.With the microsphere collected vacuum drying 6 hours at room temperature, irradiation sterilization gets final product after the sealing packing.The envelop rate of GLP-1 microsphere is 80%, particle diameter＜60 μ m.
Animal experiment: choose 16 of adult wister rats, about body weight 200g, male and female half and half, be divided into administration group and blank group at random, the an amount of microsphere of administration group subcutaneous injection (make the 160mg microsphere according to embodiment 1, contain the about 4.5mg of 6LP-1), blank group subcutaneous injection is with the normal saline of volume.Respectively at the 1st, 5,10,15,20,25,30 day the same time after the administration, give every rats by intraperitoneal injection 18mmol/kg glucose, every Mus is adopted blank blood sample earlier before the injection, gets blood at 5,10,15,20,30 and 50 minutes eye sockets then, measures the blood glucose of injection front and back.Blood sugar detection carries out with reference to the description of glucose assays test kit (glucose oxidase method, Beijing Chemical Plant).Make blood sugar concentration and the curve chart of time (seeing Fig. 1 to Fig. 7), have figure as seen, blank group lumbar injection glucose, blood glucose obviously raises after five minutes, falls after rise slowly, does not reach the ground state blood sugar concentration in the time of 50 minutes yet.The administration group can be observed the obvious functions of blood sugar effect at the 1st, 5,10,15,20,25,30 day, promptly fell back to ground state concentration in 30 minutes.Illustrate that the GLP-1 microsphere has tangible slow release and hypoglycemic activity in vivo, be used for the treatment of diabetes or controlling body weight so the present invention can be used as the slow release formulation of GLP-1.
Claims (4)
1. subcutaneous injection sustained release microsphere agents; form by substrate, medicine and protective agent thereof and polyvinyl alcohol; it is characterized in that: described substrate is polylactic acid: hydroxyacetic acid=25: 75-75: 25 polylactic acid-glycolic guanidine-acetic acid copolymer; described medicine is a glucagon-like-peptide-1; be selected from zinc carbonate, human serum albumin, trehalose and mannitol with described protective agent and through the preparation of following method:
Substrate is dissolved in organic solvent makes oil phase; concentration is 50mg-300mg/ml; get water in the formation soluble in water of appropriate amount of drug and protective agent; interior water is added above-mentioned oil phase; ultrasonic emulsification forms colostrum; the rapid dropping of colostrum prepared microsphere in the 2-6% polyvinyl alcohol water solution and through the W/O/W solvent evaporation method.
2. subcutaneous injection sustained release microsphere agents; form by substrate, medicine and protective agent thereof; it is characterized in that: described substrate is polylactic acid: hydroxyacetic acid=25: 75-75: 25 polylactic acid-glycolic guanidine-acetic acid copolymer; described medicine is a glucagon-like-peptide-1; be selected from zinc carbonate, trehalose and mannitol with described protective agent and through the preparation of following method:
Substrate is dissolved in organic solvent makes oil phase; concentration is 50mg-300mg/ml, and it is soluble in water to get appropriate amount of drug, Polyethylene Glycol and protective agent, removes Polyethylene Glycol after the lyophilization and obtains drug powder; drug powder is dispersed in the above-mentioned oil phase and through the S/O/O solvent evaporation method prepares microsphere.
3. subcutaneous injection sustained release microsphere agents; form by substrate, medicine and protective agent thereof; it is characterized in that: described substrate is polylactic acid: hydroxyacetic acid=25: 75-75: 25 polylactic acid-glycolic guanidine-acetic acid copolymer; described medicine is a glucagon-like-peptide-1; be selected from zinc carbonate, human serum albumin and trehalose with described protective agent and through the preparation of following method:
Substrate is dissolved in organic solvent makes oil phase, with appropriate amount of drug with protective agent is soluble in water obtains drug powder through atomizing freeze drying, add drug powder in the above-mentioned oil phase and through ultra-sonic dispersion, spray drying method for preparation microsphere.
4. claim 1 or the 2 or 3 described preparations application in preparation treatment diabetes or controlling body weight medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03151059 CN1289066C (en) | 2003-09-18 | 2003-09-18 | Glicetin -1 slow release microspheric preparation and its use |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03151059 CN1289066C (en) | 2003-09-18 | 2003-09-18 | Glicetin -1 slow release microspheric preparation and its use |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1524516A CN1524516A (en) | 2004-09-01 |
| CN1289066C true CN1289066C (en) | 2006-12-13 |
Family
ID=34286885
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 03151059 Expired - Fee Related CN1289066C (en) | 2003-09-18 | 2003-09-18 | Glicetin -1 slow release microspheric preparation and its use |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1289066C (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105769771A (en) * | 2014-12-25 | 2016-07-20 | 四川科伦药物研究院有限公司 | Exenatide slow-release microsphere composition and preparation method thereof |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100344323C (en) * | 2004-09-30 | 2007-10-24 | 华东师范大学 | Human glucagon-like peptide-1 compound and its preparing method |
| CN101292951B (en) * | 2007-04-23 | 2012-03-28 | 江苏先声药物研究有限公司 | Recombined human vascellum esoderma inhibin durative action preparation, preparation method and application thereof |
| CN102198103B (en) * | 2011-05-30 | 2014-07-23 | 深圳翰宇药业股份有限公司 | Stable exenatide sustained-release microsphere preparation and preparation method thereof |
| CN105878191B (en) * | 2016-04-26 | 2021-01-22 | 广州帝奇医药技术有限公司 | Preparation method of sustained-release microparticles, prepared sustained-release microparticles and application thereof |
| CN105878190B (en) * | 2016-04-26 | 2021-01-22 | 广州帝奇医药技术有限公司 | Preparation method of sustained-release microparticles, prepared sustained-release microparticles and application thereof |
| CN106074376A (en) * | 2016-06-27 | 2016-11-09 | 江苏师范大学 | A kind of glucagon-like peptide 1 slow release nanometer formulation, preparation method and application |
| WO2020219822A1 (en) * | 2019-04-24 | 2020-10-29 | Ra Pharmaceuticals, Inc. | Compositions and methods for modulating complement activity |
| KR20220104797A (en) | 2019-11-25 | 2022-07-26 | 리제너론 파마슈티칼스 인코포레이티드 | Sustained release formulation using non-aqueous emulsion |
| WO2024011218A1 (en) * | 2022-07-08 | 2024-01-11 | Brown University | Polymeric nanoparticles for long acting delivery of a peptide and methods of making and using thereof |
-
2003
- 2003-09-18 CN CN 03151059 patent/CN1289066C/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105769771A (en) * | 2014-12-25 | 2016-07-20 | 四川科伦药物研究院有限公司 | Exenatide slow-release microsphere composition and preparation method thereof |
| CN105769771B (en) * | 2014-12-25 | 2019-02-01 | 四川科伦药物研究院有限公司 | A kind of Exenatide release microsphere composition and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1524516A (en) | 2004-09-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US10363287B2 (en) | Method of manufacturing an osmotic delivery device | |
| EP1335661B1 (en) | Production of microspheres | |
| JP5175017B2 (en) | Particulate carrier for improving oral absorption of active ingredients | |
| US20070184015A1 (en) | Novel PEGylation agent | |
| CN101658496A (en) | Exenatide release microsphere preparation, preparation method and application thereof | |
| NO301405B1 (en) | Process for the preparation of extended release microcapsules | |
| CN1527698A (en) | Controlled release biodegradable gel substrate | |
| WO2017186073A1 (en) | Preparation method of sustained release microparticulates, sustained release microparticulates thereby and use thereof | |
| CN102688198A (en) | Polypeptide drug sustained-release microsphere preparation and preparation method thereof | |
| CN1289066C (en) | Glicetin -1 slow release microspheric preparation and its use | |
| CN102014880A (en) | Therapeutic calcium phosphate particles and methods of making and using same | |
| CN100345535C (en) | The Controlled release preparation of insulin and its method | |
| CN108434118A (en) | Glucagon-like peptide-1 analogs sustained-release micro-spheres and preparation method thereof | |
| CN102573799A (en) | Pharmaceutical composition containing medicament-containing fine particles and method for producing same | |
| CN1965810A (en) | Sustained release microsphere formulation of thymosin alpha-1, preparation process and use thereof | |
| US20080305139A1 (en) | Pharmaceutical Compositions Comprising Dextran with a Molecular Weight of 1.0-100 Kda and Processes for Their Preparation | |
| CN102370624A (en) | Exendin-4 sustained release microsphere, its injection and preparation method | |
| CN1302807C (en) | Biological degradable macromolecule microspheres of carrying insulin and preparation method | |
| US11246913B2 (en) | Suspension formulation comprising an insulinotropic peptide |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Assignee: Hainan Shuang Cheng Pharmaceutical Co., Ltd. Assignor: Army Medical Univ. No.2, Chinese PLA Contract fulfillment period: 2008.10.16 to 2013.10.16 Contract record no.: 2008460000017 Denomination of invention: Glicetin -1 slow release microspheric preparation and its use Granted publication date: 20061213 License type: Exclusive license Record date: 20081112 |
|
| LIC | Patent licence contract for exploitation submitted for record |
Free format text: EXCLUSIVE LICENSE; TIME LIMIT OF IMPLEMENTING CONTACT: 2008.10.16 TO 2013.10.16; CHANGE OF CONTRACT Name of requester: HAINAN SHUANGCHENG PHARMACEUTICAL CO., LTD. Effective date: 20081112 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20061213 Termination date: 20180918 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |