CN1308034C - Method of preparing microsphere of ethoxyl copolymer PLGA in interferon poly acid - Google Patents
Method of preparing microsphere of ethoxyl copolymer PLGA in interferon poly acid Download PDFInfo
- Publication number
- CN1308034C CN1308034C CNB2004100776220A CN200410077622A CN1308034C CN 1308034 C CN1308034 C CN 1308034C CN B2004100776220 A CNB2004100776220 A CN B2004100776220A CN 200410077622 A CN200410077622 A CN 200410077622A CN 1308034 C CN1308034 C CN 1308034C
- Authority
- CN
- China
- Prior art keywords
- interferon
- microsphere
- preparation
- polylactic acid
- acetic acid
- 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
- 239000004005 microsphere Substances 0.000 title claims abstract description 49
- 108010050904 Interferons Proteins 0.000 title claims abstract description 35
- 102000014150 Interferons Human genes 0.000 title claims abstract description 34
- 229920001577 copolymer Polymers 0.000 title claims abstract description 34
- 229940079322 interferon Drugs 0.000 title claims abstract description 34
- 239000002253 acid Substances 0.000 title claims description 4
- 229920001606 poly(lactic acid-co-glycolic acid) Polymers 0.000 title claims 4
- 238000000034 method Methods 0.000 title abstract description 9
- 238000002360 preparation method Methods 0.000 claims abstract description 38
- 238000004108 freeze drying Methods 0.000 claims abstract description 13
- 238000002347 injection Methods 0.000 claims abstract description 12
- 239000007924 injection Substances 0.000 claims abstract description 12
- 108010047761 Interferon-alpha Proteins 0.000 claims abstract description 7
- 102000006992 Interferon-alpha Human genes 0.000 claims abstract description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Natural products CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 40
- 239000000243 solution Substances 0.000 claims description 28
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 24
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
- 210000003022 colostrum Anatomy 0.000 claims description 18
- 235000021277 colostrum Nutrition 0.000 claims description 18
- 238000003756 stirring Methods 0.000 claims description 16
- 239000000839 emulsion Substances 0.000 claims description 14
- 208000035126 Facies Diseases 0.000 claims description 12
- 239000011780 sodium chloride Substances 0.000 claims description 12
- 230000015572 biosynthetic process Effects 0.000 claims description 11
- 239000003960 organic solvent Substances 0.000 claims description 11
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical group ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 9
- BPMFZUMJYQTVII-UHFFFAOYSA-N alpha-guanidinoacetic acid Natural products NC(=N)NCC(O)=O BPMFZUMJYQTVII-UHFFFAOYSA-N 0.000 claims description 8
- 239000007864 aqueous solution Substances 0.000 claims description 8
- 230000001804 emulsifying effect Effects 0.000 claims description 8
- 238000005406 washing Methods 0.000 claims description 8
- 239000003381 stabilizer Substances 0.000 claims description 7
- 238000004945 emulsification Methods 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 239000008307 w/o/w-emulsion Substances 0.000 claims description 4
- 108010010803 Gelatin Proteins 0.000 claims description 3
- 229920000159 gelatin Polymers 0.000 claims description 3
- 239000008273 gelatin Substances 0.000 claims description 3
- 235000019322 gelatine Nutrition 0.000 claims description 3
- 235000011852 gelatine desserts Nutrition 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 101000959794 Homo sapiens Interferon alpha-2 Proteins 0.000 claims description 2
- 229920002565 Polyethylene Glycol 400 Polymers 0.000 claims description 2
- 238000005119 centrifugation Methods 0.000 claims description 2
- JLFNLZLINWHATN-UHFFFAOYSA-N pentaethylene glycol Chemical compound OCCOCCOCCOCCOCCO JLFNLZLINWHATN-UHFFFAOYSA-N 0.000 claims description 2
- 239000003814 drug Substances 0.000 abstract description 17
- 229940079593 drug Drugs 0.000 abstract description 12
- 238000009826 distribution Methods 0.000 abstract description 5
- 239000002245 particle Substances 0.000 abstract description 5
- 239000002904 solvent Substances 0.000 abstract description 2
- 238000005538 encapsulation Methods 0.000 abstract 1
- 230000000694 effects Effects 0.000 description 7
- 238000013268 sustained release Methods 0.000 description 4
- 239000012730 sustained-release form Substances 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 3
- 239000011806 microball Substances 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000001647 drug administration Methods 0.000 description 2
- 230000003203 everyday effect Effects 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 229920000747 poly(lactic acid) Polymers 0.000 description 2
- 239000004626 polylactic acid Substances 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 102000009027 Albumins Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 102000014914 Carrier Proteins Human genes 0.000 description 1
- 108010078791 Carrier Proteins Proteins 0.000 description 1
- 229940124872 Hepatitis B virus vaccine Drugs 0.000 description 1
- 102000002265 Human Growth Hormone Human genes 0.000 description 1
- 108010000521 Human Growth Hormone Proteins 0.000 description 1
- 239000000854 Human Growth Hormone Substances 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 108010000817 Leuprolide Proteins 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 229940040591 biotech drug Drugs 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000012876 carrier material Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 239000003405 delayed action preparation Substances 0.000 description 1
- 238000001938 differential scanning calorimetry curve Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- GFIJNRVAKGFPGQ-LIJARHBVSA-N leuprolide Chemical compound CCNC(=O)[C@@H]1CCCN1C(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](CC(C)C)NC(=O)[C@@H](NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H]1NC(=O)CC1)CC1=CC=C(O)C=C1 GFIJNRVAKGFPGQ-LIJARHBVSA-N 0.000 description 1
- 229960004338 leuprorelin Drugs 0.000 description 1
- 230000001592 luteinising effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000002861 polymer material Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000009711 regulatory function Effects 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 238000000935 solvent evaporation Methods 0.000 description 1
- 150000005846 sugar alcohols Polymers 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
Images
Landscapes
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The present invention discloses a preparation method for an interferon polylactic acid-glycolate copolymer (PLGA) microsphere, which is characterized in that the microsphere is prepared by a multiple emulsion-solvent volatilization method. A polylactic acidglycolate copolymer (PLGA) is a microsphere carrier, and recombinant human interferon-alpha is taken as a packaged object in a freeze-drying injection process. The manufactured interferon microsphere has round and normal form and uniform particle size distribution. The particle size distribution is below 30 microns, the drug bearing capacity can reach more than 8%, and the encapsulation efficiency is about 40%. The extrasomatic drug release property accords with the character of a long-acting preparation.
Description
Technical field
The present invention relates to a kind of preparation method of biodegradable control-release microsphere of biologically active drug, belong to the crossing research field of biological medical polymer material and biologically active drug controlled release preparation.
Background technology
In recent years, along with the high speed development of biotechnology, increasing polypeptide, pharmaceutical grade protein are developed out, and show the curative effect of many uniquenesses in clinical practice, for example insulin, Hepatitis B virus vaccine, interferon etc.But because polypeptide and protein medicaments biological half-life are short, poor stability easily by enzymes metabolism in the body, and is difficult to absorb in gastrointestinal tract, so need for a long time, drug administration by injection continually, has hindered that it is convenient clinically, extensively and has effectively used.
Human recombinant interferon-α is the product that adopts biological high-tech preparation, by the engineering bacteria that has the human interferon gene through efficiently expressing, and the off-white powder highly purified, that lyophilization is made, it has stronger immunoloregulation function, can suppress the effect of tumor proliferation, antiviral and enhancing human body immunity regulatory function.Be mainly used in treatment rheumatoid arthritis, tumor, viral disease.Because the oral inactivation of interferon, need drug administration by injection, and the half-life in blood is short, poor stability, easily by enzymes metabolism in the body, be eliminated soon after the injection or degrade,, need frequent, heavy dose of and long-time administration in order to reach curative effect, usually usage is intramuscular injection every day 50-200 ten thousand IU, and 3 months is a course of treatment.
Utilizing the biological degradation polyalcohol microparticulate systems to come transport protein matter medicine is a main method of current domestic and international research, report is arranged, with the polylactic acid-hydroxide acetic acid copolymer is human growth hormone's microball preparation of carrier, and its drug effect than injection every day is once injected 28 days better effects if continuously.Interferon is made polylactic acid microsphere, can reach the raising medicine stability, prolong action time and purpose easy to use.The report of interferon microball preparation was abroad promptly arranged at twentieth century in 90 years, and its carrier material has albumin, gelatin and polylactic acid-hydroxide acetic acid copolymer (PLGA) etc., and the PLGA microsphere is long and more satisfactory because of drug release time.
At present, scientist is when the development sustained release microsphere agents both at home and abroad, and using maximum biodegradated polymer materals is polylactic acid-hydroxide acetic acid copolymer, because of it has favorable biological degradability, the compatibility and absorbability, is pharmaceutic adjuvant by the FDA approval.And polylactic acid sustained-release micro-spheres product such as luteinising hormone-releasing hormo, leuprorelin official listing abroad.Because polylactic acid-hydroxide acetic acid copolymer is not also produced at home as medicinal materials, and human recombinant interferon-α has how tame manufacturer production in China, price is suitable, if interferon-' alpha ' is made the polylactic acid-hydroxide acetic acid copolymer sustained release microsphere agents, to improve its added value greatly, not only can promote the exploitation of biotech drug, but also can promote the industrialization process of pharmaceutic adjuvant.
Summary of the invention
The objective of the invention is to adopt polylactic acid-hydroxide acetic acid copolymer (PLGA) is carrier, interferon-' alpha ' is made microball preparation, reach the increase medicine stability, prolong drug action time, reduce frequency injection and consumption, improve the purpose of curative effect of medication and economic benefit, and the preparation method of interferon microsphere is provided.
Interferon polylactic acid-hydroxide acetic acid copolymer of the present invention (PLGA) method for preparing microsphere is to adopt emulsion-solvent evaporated method.Polylactic acid-hydroxide acetic acid copolymer is dissolved in the organic solvent; Lyophilizing injection recombinant human interferon alpha 2 is dissolved in the interior water of formation in the PBS solution that contains stabilizing agent; Interior water is injected in the organic facies, stirring and emulsifying becomes the W/O colostrum again; In the water PVA aqueous solution, be emulsified into the W/O/W emulsion outside then colostrum being added under stirring condition; Stirring at low speed in containing electrolytical solution makes the organic solvent volatilization fully again; Centrifugal, washing, collect microsphere, lyophilization promptly gets the microsphere of ethoxyl copolymer PLGA in interferon poly acid powder.
Interferon polylactic acid-hydroxide acetic acid copolymer (PLGA) method for preparing microsphere is characterized in that preparation method may further comprise the steps:
(1) preparation of polylactic acid-hydroxide acetic acid copolymer PLGA solution
Polylactic acid-glycolic guanidine-acetic acid copolymer p LGA is dissolved in forms organic facies in the organic solvent, concentration 20%~30%g/ml of PLGA;
(2) preparation of interferon PBS solution
Lyophilizing injection recombinant human interferon-alpha is dissolved in the interior water of formation in the PBS solution that contains a certain amount of stabilizing agent, and the concentration of interferon is 13.3%~20%g/ml, and the concentration of stabilizing agent is 11%~30%g/ml;
(3) preparation of colostrum
Interior water is injected into formation W/O colostrum in the organic facies, and emulsifying rate is 1000~9000rpm, and emulsification times is 1~3 minute, and the volume ratio of organic facies and interior water is 20: 3;
(4) preparation of emulsion
Colostrum is added under condition of stirring in the outer water aqueous solution of certain density PVA and NaCl, is emulsified into the W/O/W emulsion; The mixing speed that emulsion forms is 1600~2000rpm, and the concentration of PVA solution is 1~3%g/ml, and the concentration of NaCl solution is 0.5~1%g/ml, and emulsification times is 10~20 minutes, and colostrum is 2.3: 200 to 2.3: 300 with the volume ratio of outer water;
(5) formation of microsphere
Emulsion solution is transferred in the NaCl aqueous solution, and stirring at low speed is complete, centrifugal to the organic solvent volatilization, microsphere is collected in the washing back; The concentration of NaCl solution is 1%g/ml, and volume is 2-3 a times of emulsion, and mixing speed is 1000rpm, and centrifugal speed is 1200~1500rpm, and centrifugation time is 15-20 minute, and washing times is 3 times;
(6) lyophilization.
Good effect of the present invention is: interferon polylactic acid-hydroxide acetic acid copolymer (PLGA) microsphere that the present invention adopts emulsion-solvent evaporation method to prepare, microsphere stable preparation process, feasible, the form rounding of microsphere, smooth surface, good fluidity, even particle size distribution, mean diameter is 30 μ m, drug loading is more than 8%, and envelop rate is about 40%, and its external Release Performance meets the durative action preparation feature.Interferon-' alpha ' is made sustained release microsphere agents, but prolong drug action time is improved curative effect of medication and economic benefit.
Accompanying drawing and explanation
Fig. 1 adopts the optical microscope and the electron scanning micrograph of the interferon PLGA microsphere that the present invention prepares, and observes configuration of surface.
Fig. 2 is the DSC curve of the blank microsphere (1) of PLGA, interferon PLGA medicine carrying microballoons (2).
The specific embodiment
The invention will be further described for following examples, so that those skilled in the art further understands the present invention, but do not limit the present invention in any form.
The preparation of embodiment 1 interferon polylactic acid-glycolic guanidine-acetic acid copolymer (PLGA) microsphere
(1) preparation of polylactic acid-hydroxide acetic acid copolymer (PLGA) solution
400mg polylactic acid-glycolic guanidine-acetic acid copolymer (PLGA) is dissolved in forms organic facies (20%g/ml) in the 2ml dichloromethane.
(2) preparation of interferon PBS solution
40mg lyophilizing injection recombined human interferon-alpha is dissolved in form in the gelatin solution of 300 μ l1% (g/ml) concentration in water;
(3) preparation of colostrum
Interior water is injected in the organic facies, and stirring and emulsifying 1 minute is to form colostrum (W/O) under the 9000rpm condition;
(4) preparation of emulsion
Colostrum is added to 300ml contains in 1% (g/ml) PVA and 1% (g/ml) NaCl solution (outer water) under the 2000rpm condition of stirring, emulsifying 10 minutes is emulsified into emulsion (W/O/W);
(5) formation of microsphere
Emulsion solution is transferred in 3 times of amount 1% (g/ml) NaCl aqueous solutions, and the 1000rpm stirring at low speed is complete to the organic solvent volatilization, and centrifugal 15 minutes of 1500rpm collects microsphere with distilled water wash 3 times, washing back;
(6) made microsphere is carried out lyophilization in the following manner successively: in-40 ℃, dry 4h; Follow in-25 ℃ dry 10h; Follow in-10 ℃ dry 20h; At last in-4 ℃, dry 48h.
Made microsphere is carried out lyophilization, under above-mentioned lyophilization condition, obtain the Powdered interferon polylactic acid-glycolic of white loose guanidine-acetic acid copolymer microsphere.
The form rounding of gained interferon microsphere, smooth surface, good fluidity, even particle size distribution, mean diameter are 10.71 μ m, and drug loading is 6.96%, and envelop rate is 32.90%, and external Release Performance meets the durative action preparation feature.
The preparation of embodiment 2 interferon polylactic acid-glycolic guanidine-acetic acid copolymer (PLGA) microspheres
(1) preparation of polylactic acid-hydroxide acetic acid copolymer (PLGA) solution
600mg polylactic acid-glycolic guanidine-acetic acid copolymer (PLGA) is dissolved in forms organic facies (30%g/ml) in the 2ml dichloromethane.
(2) preparation of interferon PBS solution
60mg lyophilizing injection recombined human interferon-alpha is dissolved in the interior water of formation in the 300 μ l 30%PEG400 solution;
(3) preparation of colostrum
Interior water is injected in the organic facies, and stirring and emulsifying 3 minutes is to form colostrum (W/O) under the 1000rpm condition;
(4) preparation of emulsion
Colostrum is added to 200ml contains in 3% (g/ml) PVA and 0.5% (g/ml) NaCl solution (outer water) under the 1600rpm condition of stirring, emulsifying 20 minutes is emulsified into emulsion (W/O/W);
(5) formation of microsphere
Emulsion solution is transferred in 2 times of amount 1% (g/ml) NaCl aqueous solutions, and the 1000rpm stirring at low speed is complete to the organic solvent volatilization, and centrifugal 20 minutes of 1200rpm collects microsphere with distilled water wash 3 times, washing back;
(6) made microsphere is carried out lyophilization in the following manner successively: in-40 ℃, dry 4h; Follow in-25 ℃ dry 10h; Follow in-10 ℃ dry 20h; At last in-4 ℃, dry 48h.
Made microsphere is carried out lyophilization, under above-mentioned lyophilization condition, obtain the Powdered interferon polylactic acid-glycolic of white loose guanidine-acetic acid copolymer microsphere.
The form rounding of gained interferon microsphere, smooth surface, good fluidity, even particle size distribution, mean diameter are 12.36 μ m, and drug loading is 6.06%, and envelop rate is 36.80%, and external Release Performance meets the durative action preparation feature.
Claims (5)
1, a kind of preparation method of interferon polylactic acid-hydroxide acetic acid copolymer microsphere, carry out according to the following steps:
Polylactic acid-hydroxide acetic acid copolymer is dissolved in the organic solvent; Lyophilizing injection recombinant human interferon alpha 2 is dissolved in the interior water of formation in the PBS solution that contains stabilizing agent; Interior water is injected in the organic facies, stirring and emulsifying becomes the W/O colostrum again; In the water PVA aqueous solution, be emulsified into the W/O/W emulsion outside then colostrum being added under stirring condition; Stirring at low speed in containing electrolytical solution makes the organic solvent volatilization fully again; Centrifugal, washing, collect microsphere, lyophilization promptly gets the microsphere of ethoxyl copolymer PLGA in interferon poly acid powder.
2, interferon polylactic acid-hydroxide acetic acid copolymer microsphere preparation method as claimed in claim 1, it is characterized in that: preparation method may further comprise the steps:
(1) preparation of polylactic acid-hydroxide acetic acid copolymer PLGA solution
Polylactic acid-glycolic guanidine-acetic acid copolymer p LGA is dissolved in forms organic facies in the organic solvent, concentration 20%~30%g/ml of PLGA;
(2) preparation of interferon PBS solution
Lyophilizing injection recombinant human interferon-alpha is dissolved in the interior water of formation in the PBS solution that contains a certain amount of stabilizing agent, and the concentration of interferon is 13.3%~20%g/ml, and the concentration of stabilizing agent is 11%~30%g/ml;
(3) preparation of colostrum
Interior water is injected into formation W/O colostrum in the organic facies, and emulsifying rate is 1000~9000rpm, and emulsification times is 1~3 minute, and the volume ratio of organic facies and interior water is 20: 3;
(4) preparation of emulsion
Colostrum is added under condition of stirring in the outer water aqueous solution of certain density PVA and NaCl, is emulsified into the W/O/W emulsion; The mixing speed that emulsion forms is 1600~2000rpm, and the concentration of PVA solution is 1~3%g/ml, and the concentration of NaCl solution is 0.5~1%g/ml, and emulsification times is 10~20 minutes, and colostrum is 2.3: 200 to 2.3: 300 with the volume ratio of outer water;
(5) formation of microsphere
Emulsion solution is transferred in the NaCl aqueous solution, and stirring at low speed is complete, centrifugal to the organic solvent volatilization, microsphere is collected in the washing back; The concentration of NaCl solution is 1%g/ml, and volume is 2-3 a times of emulsion, and mixing speed is 1000rpm, and centrifugal speed is 1200~1500rpm, and centrifugation time is 15-20 minute, and washing times is 3 times;
(6) lyophilization.
3. interferon polylactic acid-hydroxide acetic acid copolymer microsphere preparation method as claimed in claim 1 or 2 is characterized in that: described organic solvent is a dichloromethane.
4. interferon polylactic acid-hydroxide acetic acid copolymer microsphere preparation method as claimed in claim 1 or 2 is characterized in that: the stabilizing agent of described interferon is gelatin or PEG400.
5. interferon polylactic acid-hydroxide acetic acid copolymer microsphere preparation method as claimed in claim 1 or 2 is characterized in that: made microsphere is carried out lyophilization in the following manner successively: in-40 ℃, and dry 4h; Follow in-25 ℃ dry 10h; Follow in-10 ℃ dry 20h; At last in-4 ℃, dry 48h.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CNB2004100776220A CN1308034C (en) | 2004-12-27 | 2004-12-27 | Method of preparing microsphere of ethoxyl copolymer PLGA in interferon poly acid |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CNB2004100776220A CN1308034C (en) | 2004-12-27 | 2004-12-27 | Method of preparing microsphere of ethoxyl copolymer PLGA in interferon poly acid |
Publications (2)
Publication Number | Publication Date |
---|---|
CN1660412A CN1660412A (en) | 2005-08-31 |
CN1308034C true CN1308034C (en) | 2007-04-04 |
Family
ID=35010149
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CNB2004100776220A Expired - Fee Related CN1308034C (en) | 2004-12-27 | 2004-12-27 | Method of preparing microsphere of ethoxyl copolymer PLGA in interferon poly acid |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN1308034C (en) |
Families Citing this family (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN100460441C (en) * | 2006-09-25 | 2009-02-11 | 南开大学 | Organism degradable star-type structure poly (glycolide-lactide) medicine carrier microsphere and preparation method thereof |
CN100457187C (en) * | 2006-11-10 | 2009-02-04 | 中国人民解放军第二军医大学 | VEGF slowly releasing injection microsphere support and its prepn and use |
UY33517A (en) * | 2010-07-19 | 2012-02-29 | Astrazeneca Ab | Pharmaceutical depot for 5-fluoro-2 - [[(1S) -1- (5-fluoro-2-pyridyl) ethyl] amino] -6 - [(5-isopropoxy-1H-pyrazol-3-yl) amino] pyridin -3-carbonitrile ?. |
CN107281111A (en) * | 2016-04-05 | 2017-10-24 | 南方医科大学南方医院 | A kind of degradable polymer contains the preparation method of NBD polypeptide microballoons |
CN107698795B (en) * | 2017-09-30 | 2020-10-27 | 西安交通大学 | Preparation method and application of porous polymer microspheres with controllable structures |
CN109172874A (en) * | 2018-09-18 | 2019-01-11 | 福建师范大学 | A method of preparing the polylactic acid microsphere of gelatin surface graft modification |
CN114634634A (en) * | 2022-03-22 | 2022-06-17 | 陈凌卉 | Biological function composite porous polyester microsphere and preparation method thereof |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1140058A (en) * | 1996-03-29 | 1997-01-15 | 中国科学院成都有机化学研究所 | Slow-release microsphere powder injection of androgenic hormone, its preparing method and use |
BR0300713A (en) * | 2003-03-24 | 2004-11-16 | Ct Ingenieria Genetica Biotech | Immunogenic compositions for prevention and treatment against endoparasites and ectoparasites |
-
2004
- 2004-12-27 CN CNB2004100776220A patent/CN1308034C/en not_active Expired - Fee Related
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1140058A (en) * | 1996-03-29 | 1997-01-15 | 中国科学院成都有机化学研究所 | Slow-release microsphere powder injection of androgenic hormone, its preparing method and use |
BR0300713A (en) * | 2003-03-24 | 2004-11-16 | Ct Ingenieria Genetica Biotech | Immunogenic compositions for prevention and treatment against endoparasites and ectoparasites |
Also Published As
Publication number | Publication date |
---|---|
CN1660412A (en) | 2005-08-31 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN1055618C (en) | Prolonged release microcapsules | |
CN1298386C (en) | Prepn process of slow release parathyroid hormone microballoon | |
AU2004259208B2 (en) | Method for the preparation of controlled release formulations | |
CN1205248C (en) | Biodegradable poly (alkylene oxide)-poly cp-dioxanone) block copolymer soluble in organic solvents, and drug delivery composition comprising same | |
CN1507357A (en) | Method and compositions for enhanced delivery of bioactive molecules | |
US20080241267A1 (en) | Hydrogel Microspheres with Improved Release Profile | |
US20070092574A1 (en) | Controlled released compositions | |
EP0946169A2 (en) | Method of producing a sustained-release preparation | |
CN1104488A (en) | Sustained releasable parenteral pharmaceutical preparations and method of producing the same | |
WO2013189282A1 (en) | Polypeptide-medicine-slow-releasing microsphere preparation and preparation method therefor | |
CN102727899A (en) | Protein-medicament-carrying PLGA composite microspheres and preparation method thereof | |
CN1308034C (en) | Method of preparing microsphere of ethoxyl copolymer PLGA in interferon poly acid | |
KR20110076783A (en) | A polymer for protein, polypeptide or peptide drug delivery and a method for preparing the same, and a composition for sustained release of protein, polypeptide or peptide drug and a method for preparing the same | |
CN100345535C (en) | The Controlled release preparation of insulin and its method | |
CN1604788A (en) | High-concentration protein formulations and method of manufacture | |
CN1823740A (en) | Taxadol slow release nano-particle, its preparation method and application | |
CN1907270A (en) | Method for preparing protein-polysaccharide vitreum slow release microsphere by using low-temperature aqueous-aqueous phase emulsion | |
CN104984327A (en) | Thymalfasin sustained release micro-sphere preparation and preparing method thereof | |
CN1887273A (en) | Prepn process of polysaccharide vitreous particle | |
CN1876173A (en) | Thymus pentapeptide slow-release microshpere formulation for injection and its preparation method | |
CN101947206B (en) | Method for preparing recombinant pancreotropic hormone secretion peptide medicament microspheres | |
CN1872334A (en) | Slow releasing microsphere preparation of Thymopentin, and prepartion method | |
KR101298219B1 (en) | A novel preparation method of a microsphere with high entrapment efficiency | |
CN1973843A (en) | Degradable polymer supported nanometer Daunorubicin microsphere and its prepn process | |
CN101040847A (en) | Nanometer medicine agent produced by hydrogenated castor oil and the technique of preparing the same |
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 | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20070404 Termination date: 20151227 |
|
EXPY | Termination of patent right or utility model |