CN107281111A - A kind of degradable polymer contains the preparation method of NBD polypeptide microballoons - Google Patents
A kind of degradable polymer contains the preparation method of NBD polypeptide microballoons Download PDFInfo
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- CN107281111A CN107281111A CN201610211199.1A CN201610211199A CN107281111A CN 107281111 A CN107281111 A CN 107281111A CN 201610211199 A CN201610211199 A CN 201610211199A CN 107281111 A CN107281111 A CN 107281111A
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- nbd
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- degradable polymer
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/08—Peptides having 5 to 11 amino acids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
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- Life Sciences & Earth Sciences (AREA)
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- Proteomics, Peptides & Aminoacids (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Inorganic Chemistry (AREA)
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The present invention relates to the preparation method that a kind of degradable polymer contains NBD polypeptide microballoons.Preparation method of the present invention comprises the following steps：The water-in-oil emulsion being made up of the organic solution of the good degradable polymer of biocompatibility and the aqueous solution of NBD polypeptides, the colostric fluid after being emulsified as first time；Colostric fluid is injected in syringe, promotes syringe colostric fluid is injected into PVA shearing liquid with certain speed with medical micro-injection pump, is transferred in centrifuge tube, is placed in mixer and stirs；Drive the iron rod being fixed thereon to rotate together when mixer is rotated with certain speed, drive PVA solution rotation to produce shear action, colostric fluid is gradually elongated under the shearing force of rotating liquid to attenuate, and is separated into small ball；The small ball of emulsion formation gradually spreads in PVA solution, is solidified into microballoon of uniform size；By microballoon centrifuge washing, the microballoon being collected into is freeze-dried.The present invention can prepare that particle size is homogeneous, the stable microballoon of envelop rate, and preparation technology is more stable.
Description
Technical field
The invention belongs to biomedical sector, it is related to a kind of preparation method of medicine microspheres, it is more particularly to a kind of
Degradable polymer contains the preparation method of NBD polypeptide microballoons.
Background technology
In Cranial defect treatment clinical course, the presence of persistent inflammation and infection will cause Postoperative Bone focus of infection
Recurrence so that the failure of debridement surgical.Inflammatory factor TNF-α plays key player in most of inflammation,
NF- к B are crucial transcription factors in inflammatory reaction simultaneously.NBD polypeptides are a kind of with cell membrane penetration
Polypeptide, be used as inflammation inhibitor energy specific inhibition NF- к B inflammatory signals paths.But for NBD polypeptides
Intervene in bone and infect, still there is problems with to have to be solved：(1) administration of the NBD polypeptides in infection of bone focus
Mode, administration concentration and use time have to be determined；(2) half-life short in NBD polypeptides body, it is local to answer
With metabolism is diluted quickly, as the key of postoperative anti-inflammatory, how to extend its continuous action time in art area,
Reducing its first pass effect to greatest extent has to be solved simultaneously.
Biodegradable high molecular sustained-release micro-spheres delivery system has the protective effect of drug and control release of uniqueness special
Property, as polypeptide, the transport vehicle of protein-based macromolecular drug, in the activity of extension medicine, improve medicine
In terms of stability, with superiority more obvious than general pharmaceutical preparation.
The preparation method of conventional drug bearing microsphere is multi-emulsion method (w/o/w).This method mainly includes four steps：
The preparation of colostrum, the formation of emulsion, the solidification of microballoon and the collection of microballoon and washing.When multi-emulsion method is research
Between earliest, a kind of application most wide method for preparing microsphere, its technique is simple, and the microballoon drugloading rate of preparation exists
Distribution uniform in microballoon.
But, preparing micro spheres by multiple emulsion process traditionally easily causes microspherulite diameter distributing inhomogeneity, envelop rate
It is unstable, the problems such as microballoon being made is easily broken deformation, this can cause a large amount of losses of medicine, this to including
Costly medicine including NBD polypeptides, is a kind of serious loss.
Therefore, it is necessary to develop the loading microballoon of new load NBD polypeptides.
The content of the invention
In order to solve the shortcoming and defect of prior art, it is an object of the invention to provide a kind of degradable polymer
The preparation method of NBD polypeptide microballoons is contained, can prepare that particle size is homogeneous, the stable microballoon of envelop rate, and
Preparation technology is more stable, and the microballoon of various patterns can be obtained as needed.
Degradable polymer of the present invention contains the preparation method of NBD polypeptide microballoons, comprises the following steps：
A. by the organic solution and the aqueous solution group of NBD polypeptides of the good degradable polymer of biocompatibility
Into water-in-oil emulsion, be used as the first time emulsify after colostric fluid；
B. the colostric fluid is injected in syringe, the note is promoted with certain speed with medical micro-injection pump
Emitter makes the colostric fluid be injected into the PVA shearing liquid that concentration is 1%~6%, is transferred in centrifuge tube, is placed in
Stirred in mixer, in syringe and the additional ice bath of container for containing shearing liquid in whipping process, to ensure NBD
Polypeptide consistency；The iron rod being fixed thereon is driven to rotate together when mixer is rotated with certain speed, so as to drive
PVA solution rotation produces shear action, and colostric fluid is gradually elongated under the shearing force of rotating liquid to attenuate,
It is separated into small ball；The small ball of emulsion formation gradually spreads in PVA solution, and final curing is into size
Uniform microballoon；
C. after colostric fluid injection finishes reaction completion, microballoon centrifuge washing freezes the microballoon being collected into
Dry.
The further spy of the preparation method of NBD polypeptide microballoons is contained according to degradable polymer of the present invention
Levy, in the step A, the degradable polymer is to be selected from：Poly- hydroxyl ethanol sour (PLGA), gather oneself
Lactone (PCL), PLA (PLA), glycolic acid polymer (GA).
The further spy of the preparation method of NBD polypeptide microballoons is contained according to degradable polymer of the present invention
Levy, in the step A, the organic solvent in the organic solution is to be selected from：Dichloromethane (DCM), three
Vinyl chloride (TCE), tetrachloro-ethylene (PCE), tetrahydrofuran (THF), acetone or ethyl acetate (EAC).
The further spy of the preparation method of NBD polypeptide microballoons is contained according to degradable polymer of the present invention
Levy, in the step A, the organic solution of described degradable polymer is that poly- hydroxyl ethanol is sour (PLGA)
Dichloromethane (DCM) solution.Preferably, the concentration of the DCM solution of the PLGA is 2.5%~15%.
The further spy of the preparation method of NBD polypeptide microballoons is contained according to degradable polymer of the present invention
Levy, in the step A, the concentration of the aqueous solution of the NBD polypeptides is 1.24%~5%.
The further spy of the preparation method of NBD polypeptide microballoons is contained according to degradable polymer of the present invention
Levy, in the step B, syringe is fixed on medical micro-injection pump, after syringe needle is connected with syringe
Stretch into 50ml centrifuge tubes, iron rod is fixed on mechanical agitator, while iron rod is stretched into 50ml centrifuge tubes,
On horizontal level, iron rod bottom is less than needle point 1-1.5cm.
The further spy of the preparation method of NBD polypeptide microballoons is contained according to degradable polymer of the present invention
Levy, needle tubing internal diameter is 0.2~1mm, and the stirring rod distance of syringe needle and mixer is 0.1-0.3mm, and syringe needle is stretched into
0.5-5cm below PVA solution liquid level.
The further spy of the preparation method of NBD polypeptide microballoons is contained according to degradable polymer of the present invention
Levy, the rotating speed of mixer is 1600-2000rad/min.
The further spy of the preparation method of NBD polypeptide microballoons is contained according to degradable polymer of the present invention
Levy, medical micro-injection pump promotes the volume fltting speed of syringe injection to be 10-20mL/h.
Degradable polymer of the present invention contains the preparation method of NBD polypeptide microballoons, by for the first time
After emulsification, colostric fluid is injected into by the mode of the emulsion formation of abandoning tradition using syringe pump pushing syringe
In the PVA shearing liquid of low concentration, and colostrum drop is dispersed into microballoon by shearing force.Therefore shorten
Microballoon molding time and hardening time.Especially for NBD polypeptides this classes is soluble in water and half-life short
Active medicine, the time of contact of NBD polypeptides and water is not only shortened using preparation method of the present invention,
And the precipitation of NBD polypeptides can be prevented, so as to be effectively improved the envelop rate and drugloading rate of microballoon.Cause
This, method for preparing microsphere of the present invention is not only effectively improved the envelop rate and carrying drug ratio of microballoon, and substantially
Reduce the waste of NBD polypeptides.
Compared with traditional microsphere preparation technology, preparation method of the present invention has the advantage that：(1) shorten
In the reaction time, reduce drug wastage；(2) syringe pump fltting speed uniform, controllable, and due to stirring rod and syringe needle
Distance keeps certain in the reaction, makes shearing force suffered by colostrum drop homogeneous, thus the shaping microballoon size obtained
It is homogeneous.
Brief description of the drawings
Fig. 1 is that degradable polymer of the present invention contains what is used in the preparation method of NBD polypeptide microballoons
Injection and the structural representation of agitating device.
Fig. 2 is the optical imagery using the NBD polypeptide microballoons prepared by preparation method of the present invention.
Fig. 3 is the scanning electron microscope image using the NBD polypeptide microballoons prepared by preparation method of the present invention.
Embodiment
Combining appended accompanying drawing below by specific embodiment, the present invention is described in further detail, but should not
It is considered as limitation of the present invention.
Degradable polymer of the present invention contains the preparation method of NBD polypeptide microballoons, comprises the following steps：
A. by the organic solution and the aqueous solution group of NBD polypeptides of the good degradable polymer of biocompatibility
Into water-in-oil emulsion, be used as the first time emulsify after colostric fluid；
B. the colostric fluid is injected in syringe, the note is promoted with certain speed with medical micro-injection pump
Emitter makes the colostric fluid be injected into the PVA shearing liquid that concentration is 1%~6%, is transferred in centrifuge tube, is placed in
Stirred in mixer, in syringe and the additional ice bath of container for containing shearing liquid in whipping process, to ensure NBD
Polypeptide consistency；The iron rod being fixed thereon is driven to rotate together when mixer is rotated with certain speed, so as to drive
PVA solution rotation produces shear action, and colostric fluid is gradually elongated under the shearing force of rotating liquid to attenuate,
It is separated into small ball；The small ball of emulsion formation gradually spreads in PVA solution, and final curing is into size
Uniform microballoon；
C. after colostric fluid injection finishes reaction completion, microballoon centrifuge washing freezes the microballoon being collected into
Dry.
As shown in figure 1, the preparation method that degradable polymer of the present invention contains NBD polypeptide microballoons is adopted
With following device：Mixer 1, iron rod 2, medical micro-injection pump 3, syringe 4, container 5.Iron rod
2 are fixed on mixer 1, can be rotated under the drive of mixer 1.Colostric fluid is by medical micro-injection
In container 5 of the pushing syringe 4 of pump 3 injection equipped with PVA solution, iron rod 2 is under the drive of mixer 1
Rotation, thus drives PVA solution rotation to produce shear action.During stirring, medical micro-injection pump 3 with
The outside bath 6 on the rocks of the container 5 of shearing liquid is contained, to ensure NBD polypeptide consistency.
Colostric fluid is gradually elongated under the shearing force of rotating liquid to attenuate, and is separated into small ball.Emulsion shape
Into small ball gradually spread in PVA solution, final curing is into microballoon of uniform size.React completion
Afterwards, centrifuge washing, freeze-drying.
Preferably, centrifugal washing times are 3-5 times.
Preferably, microballoon first pre-freeze 24-72 hours, then be freeze-dried 24-36 hours.
Embodiment 1
The poly- hydroxyl ethanols of 0.5g sour (PLGA) (Mw=50000) are dissolved in 10mL dichloromethane (DCM)
In, magnetic agitation obtains the solution that mass fraction is 5% for 3 hours, carries out deaeration processing.By 50mg NBD
Polypeptide is mixed static in 1ml water.Both the above solution is mixed, emulsified, Water-In-Oil colostrum is obtained
Liquid.It is subsequently added in syringe 4, syringe 4 is placed on medical micro-injection pump 3, sets and promote speed
Spend for 10mL/h.The concentration for shearing liquid is 2% PVA solution, and mixer rotating speed is set to 1600rad/min.
Colostric fluid balling-up under shear action.Obtained microballoon centrifuge washing, freeze-drying.
Poly- hydroxyl ethanol sour (PLGA) in the present embodiment can be substituted with following degradable polymer：Gather oneself
Lactone (PCL), PLA (PLA), glycolic acid polymer (GA).
Dichloromethane (DCM) in the present embodiment can be substituted with following organic solvent：Trichloro ethylene (TCE),
Tetrachloro-ethylene (PCE), tetrahydrofuran (THF), acetone or ethyl acetate (EAC).
Embodiment 2
The poly- hydroxyl ethanols of 0.25g sour (PLGA) (Mw=50000) are dissolved in 10mL dichloromethane (DCM)
In, magnetic agitation obtains the solution that mass fraction is 2.5% for 3 hours, carries out deaeration processing.By 50mg NBD
Polypeptide is mixed static in 1ml water.Both the above solution is mixed, emulsified, Water-In-Oil colostrum is obtained
Liquid.It is subsequently added in syringe 4, syringe 4 is placed on medical micro-injection pump 3, fltting speed is set
For 10mL/h.The concentration for shearing liquid is 2% PVA solution.Mixer rotating speed is set to 1600rad/min.
The balling-up under shear action.Obtained microballoon centrifuge washing, freeze-drying.
Poly- hydroxyl ethanol sour (PLGA) in the present embodiment can be substituted with following degradable polymer：Gather oneself
Lactone (PCL), PLA (PLA), glycolic acid polymer (GA).
Dichloromethane (DCM) in the present embodiment can be substituted with following organic solvent：Trichloro ethylene (TCE),
Tetrachloro-ethylene (PCE), tetrahydrofuran (THF), acetone or ethyl acetate (EAC).
Embodiment 3
The poly- hydroxyl ethanols of 1.5g sour (PLGA) (Mw=50000) are dissolved in 10mL dichloromethane (DCM)
In, magnetic agitation obtains the solution that mass fraction is 15% for 3 hours, carries out deaeration processing.By 50mg NBD
Polypeptide is mixed static in 1ml water.Both the above solution is mixed, emulsified, Water-In-Oil colostrum is obtained
Liquid.It is subsequently added in syringe 4, syringe 4 is placed on medical micro-injection pump 3, fltting speed is set
For 20mL/h.The concentration for shearing liquid is 2% PVA solution.Mixer rotating speed is set to 2000rad/min.
The balling-up under shear action.Obtained microballoon centrifuge washing, freeze-drying.
Poly- hydroxyl ethanol sour (PLGA) in the present embodiment can be substituted with following degradable polymer：Gather oneself
Lactone (PCL), PLA (PLA), glycolic acid polymer (GA).
Dichloromethane (DCM) in the present embodiment can be substituted with following organic solvent：Trichloro ethylene (TCE),
Tetrachloro-ethylene (PCE), tetrahydrofuran (THF), acetone or ethyl acetate (EAC).
Embodiment 4
The NBD polypeptide microballoon observation by light microscope that will be prepared according to embodiment 1, as a result as shown in Fig. 2
Microspherulite diameter size is homogeneous, and envelop rate is stable.It is scanned with SEM (SEM), as a result such as
Shown in Fig. 3, the microspherulite diameter prepared with this kind of method is more uniform.
Above-described embodiment is preferably embodiment, but embodiments of the present invention are not by above-mentioned implementation of the invention
The change made under the limitation of example, other any Spirit Essences and principle without departing from the present invention, modify, replace
Generation, combination, simplification, should be equivalent substitute mode, are included within protection scope of the present invention.
Claims (10)
1. a kind of degradable polymer contains the preparation method of NBD polypeptide microballoons, it is characterised in that the preparation side
Method comprises the following steps：
A. it is made up of the organic solution of the good degradable polymer of biocompatibility and the aqueous solution of NBD polypeptides
Water-in-oil emulsion, the colostric fluid after being emulsified as first time；
B. the colostric fluid is injected in syringe, the syringe is promoted with certain speed with medical micro-injection pump
The colostric fluid is injected into the PVA shearing liquid that concentration is 1%~6%, be transferred in centrifuge tube, be placed in stirring
Stirred in machine, in syringe and the additional ice bath of container for containing shearing liquid in whipping process, to ensure that NBD is more
Peptide consistency；The iron rod being fixed thereon is driven to rotate together when mixer is rotated with certain speed, so as to drive
PVA solution rotation produces shear action, and colostric fluid is gradually elongated under the shearing force of rotating liquid to attenuate,
It is separated into small ball；The small ball of emulsion formation gradually spreads in PVA solution, and final curing is into size
Uniform microballoon；
C. after colostric fluid injection finishes reaction completion, the microballoon being collected into is freeze-dried by microballoon centrifuge washing.
In syringe and the additional ice bath of container for containing shearing liquid in whipping process, to ensure NBD polypeptide consistency.
2. degradable polymer according to claim 1 contains the preparation method of NBD polypeptide microballoons, its feature
It is：In the step A, the degradable polymer is to be selected from：It is poly- hydroxyl ethanol sour (PLGA), poly-
Caprolactone (PCL), PLA (PLA), glycolic acid polymer (GA).
3. degradable polymer according to claim 1 contains the preparation method of NBD polypeptide microballoons, its feature
It is：In the step A, the organic solvent in the organic solution is to be selected from：Dichloromethane (DCM),
Trichloro ethylene (TCE), tetrachloro-ethylene (PCE), tetrahydrofuran (THF), acetone or ethyl acetate (EAC).
4. degradable polymer according to claim 1 contains the preparation method of NBD polypeptide microballoons, its feature
It is：In the step A, the organic solution of described degradable polymer is that poly- hydroxyl ethanol is sour (PLGA)
Dichloromethane (DCM) solution.
5. degradable polymer according to claim 4 contains the preparation method of NBD polypeptide microballoons, its feature
It is：The concentration of the DCM solution of the PLGA is 2.5%~15%.
6. degradable polymer according to claim 1 contains the preparation method of NBD polypeptide microballoons, its feature
It is：In the step A, the concentration of the aqueous solution of the NBD polypeptides is 1.24%~5%.
7. degradable polymer according to claim 1 contains the preparation method of NBD polypeptide microballoons, its feature
It is：In the step B, syringe is fixed on medical micro-injection pump, syringe needle is connected with syringe
After stretch into 50ml centrifuge tubes, iron rod is fixed on mechanical agitator, while iron rod stretches into 50ml centrifuge tubes
In, on horizontal level, iron rod bottom is less than needle point 1-1.5cm.
8. degradable polymer according to claim 1 contains the preparation method of NBD polypeptide microballoons, its feature
It is：Needle tubing internal diameter is 0.2~1mm, and the stirring rod distance of syringe needle and mixer is 0.1-0.3mm, and syringe needle is stretched
Enter 0.5-5cm below PVA solution liquid level.
9. degradable polymer according to claim 1 contains the preparation method of NBD polypeptide microballoons, its feature
It is：The rotating speed of mixer is 1600-2000rad/min.
10. degradable polymer according to claim 1 contains the preparation method of NBD polypeptide microballoons, it is special
Levy and be：Medical micro-injection pump promotes the volume fltting speed of syringe injection to be 10-20mL/h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201610211199.1A CN107281111A (en) | 2016-04-05 | 2016-04-05 | A kind of degradable polymer contains the preparation method of NBD polypeptide microballoons |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610211199.1A CN107281111A (en) | 2016-04-05 | 2016-04-05 | A kind of degradable polymer contains the preparation method of NBD polypeptide microballoons |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108704578A (en) * | 2018-04-20 | 2018-10-26 | 中国药科大学 | A kind of equipment of continuous production microballoon and its application |
| CN111568878A (en) * | 2020-05-22 | 2020-08-25 | 浙江圣兆药物科技股份有限公司 | Method for preparing polypeptide drug microspheres based on submerged airflow spraying technology |
| CN113244108A (en) * | 2021-06-04 | 2021-08-13 | 胡振华 | Preparation method and preparation device of polymer microspheres |
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| CN1660412A (en) * | 2004-12-27 | 2005-08-31 | 中山大学 | A kind of preparation method of interferon polylactic acid-glycolic acid copolymer PLGA microsphere |
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| CN1660412A (en) * | 2004-12-27 | 2005-08-31 | 中山大学 | A kind of preparation method of interferon polylactic acid-glycolic acid copolymer PLGA microsphere |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108704578A (en) * | 2018-04-20 | 2018-10-26 | 中国药科大学 | A kind of equipment of continuous production microballoon and its application |
| CN111568878A (en) * | 2020-05-22 | 2020-08-25 | 浙江圣兆药物科技股份有限公司 | Method for preparing polypeptide drug microspheres based on submerged airflow spraying technology |
| CN111568878B (en) * | 2020-05-22 | 2022-03-25 | 浙江圣兆药物科技股份有限公司 | Method for preparing polypeptide drug microspheres based on submerged airflow spraying technology |
| CN113244108A (en) * | 2021-06-04 | 2021-08-13 | 胡振华 | Preparation method and preparation device of polymer microspheres |
| CN113244108B (en) * | 2021-06-04 | 2023-08-25 | 胡振华 | Method and device for preparing polymer microsphere |
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