CN106913525A - It is a kind of to carry the preparation method for promoting bone synthetic drug nano particle lipopolymer - Google Patents
It is a kind of to carry the preparation method for promoting bone synthetic drug nano particle lipopolymer Download PDFInfo
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Abstract
The invention discloses a kind of preparation method for carrying and promoting bone synthetic drug nano particle lipopolymer, the sclerotin mineralising difference in the bon e formation area that the bone information area and Gegenbaur's cell gathered using osteoclast in BMU are gathered, design a kind of nano target medicine transmission system of specific selectivity, with external aspartic acid, the liposome of serine repetitive sequence DSS6, PLGA and lecithin are raw material, and add promotion bone synthetic drug, synthesis is carried and promotees bone synthetic drug nano particle lipopolymer, the nano particle for loading rush bone synthetic drug has special affinity with the low hydroxyapatite of mineralization degree, the bon e formation area that distribution of specific is assembled in Gegenbaur's cell after into bone tissue, can greatly reduce and promote the side effect that bone synthetic drug promotes bone information.Nano medicament carrying system has the advantages such as itself long circulating, targeting, slow controlled release, can overcome the defects such as existing rush bone synthetic drug bioavailability is low, stability is poor, pharmacological action time is short.
Description
Technical field
The present invention relates to field of medicaments, a kind of more particularly to load promotees bone synthetic drug nano particle fat-polymerization
The preparation method of thing.
Background technology
Osteoporosis is a kind of gradual bone information and the unbalance systemic bone disease of bon e formation, osteoporosis hair
Raw Cytological Basis are relatively weak osteoblast activity and/or stronger osteoclast activities, result in Bone m etabolism mistake
Weighing apparatus.Current osteoporosis therapy medicine can be divided into two major classes, anti-bone information medicine and rush bone synthetic drug, anti-bone information medicine
The Main Function mechanism of thing is, with osteoclast as target, to suppress pathologic bone information.And promote bone synthetic drug with Gegenbaur's cell
It is target, mainly promotes bone anabolism and bon e formation, increases bone volume and bone density.However, most of anti-bone information medicines
Thing has suppression sclerotin to synthesize, and promoting bone synthetic drug has the side effect of non-specific promotion bone information.In order to obtain more preferable curative effect, one
A little scholars also attempt the anti-bone information medicine of use in conjunction and promote bone synthetic drug, but drug combination might not have than single medicine
There is a more preferable curative effect, be the reason for produce this phenomenon：Either promote bone synthetic drug or anti-bone absorption medicine, medicine exists
It is non-distribution of specific in bone tissue, medicine can simultaneously act on Gegenbaur's cell and osteoclast, inevitably bring
Side effect.Current bone target medicine transmission system is only capable of realizing that bone tissue is targetted, and can not realize medicine in bone tissue
Accurately cell-targeting transmission.
The absorption and reconstruction of bone are a processes for dynamic equilibrium, and bone remodeling process is betided in the BMU of all bones, including
Activation, bone information, four different phases of reversion and bon e formation and constantly circulation generation, inhale so as to form obvious bone in BMU
Receive area and bon e formation area.The spatial distribution of Gegenbaur's cell and osteoclast in BMU is significantly different.It is a large amount of broken in bone information area
Osteocyte is attached to ripe bone surface dissolves bone；In bon e formation area, Gegenbaur's cell turns into leading cell and forms new bone.In bone
Uptake zone is ripe sclerotin, and mineralization degree is high, and hydroxylapatite crystal degree is high；And be newborn sclerotin, mineralising in bon e formation area
Low degree, hydroxylapatite crystal degree is low.Some special peptides show selective suction to different crystallinity hydroxyapatite
Attached characteristic, aspartic acid, serine repetitive sequence can specific adsorption in low-crystallinity hydroxyapatite surface；Asparagus fern in vitro
Propylhomoserin, serine repetitive sequence also show that the characteristic for being adsorbed to osteoblast mineralization tubercle and amorphous calcium phosphate well.
Promote the skeletonization that bone synthetic drug can be gathered with specific effect under aspartic acid, the guiding of serine repetitive sequence in bon e formation area
Cell.Therefore, aspartic acid, serine repetitive sequence can be realized promoting the thin of bone synthetic drug as selectively targeted molecule
Born of the same parents' targeted delivery.
The content of the invention
In view of the shortcomings of the prior art, carried it is an object of the invention to provide one kind and promote bone synthetic drug nano particle
The preparation method of fat-polymer, makes rush bone synthetic drug to make with specificity under aspartic acid, the guiding of serine repetitive sequence
For the Gegenbaur's cell that bon e formation area gathers, can greatly reduce and promote the side effect that bone synthetic drug promotes bone information.
To achieve the above object, the invention provides following technical scheme：One kind load rush bone synthetic drug nano particle fat-
The preparation method of polymer,
Step one, using polypeptide solid-state reaction method prepare SDSSDC；
Step 2, the external SDSSDC on liposome, obtain SDSSDC- liposomes；
Step 3, promoting bone synthetic drug, Poly(D,L-lactide-co-glycolide PLGA, lecithin and SDSSDC- liposomes
Promote bone synthetic drug nano particle fat-polymer for raw material prepares to carry.
As a further improvement on the present invention, the liposome is PEG-DSPE-Malaysia
Acid imide copolymerization, PEG-DSPE-succinimide copolymerization, DSPE-
Polyethylene glycol-silane copolymer, PEG-DSPE-amino copolymerization or distearoylphosphatidyl ethanol
One kind of amine-polyethylene glycol copolymerization.
As a further improvement on the present invention, comprising the following steps that for SDSSDC- liposomes is synthesized in the step 2：
Using 1- ethyls -3- (3- dimethyl aminopropyls)-carbodiimides and N-hydroxy-succinamide as coupling reagent
Activation SDSSDC c-terminus 3-4h, add liposome, at room temperature hybrid reaction 12-24h, dialyse, and freeze, stand-by.
As a further improvement on the present invention, the SDSSDC and the mass ratio of liposome are 1：10-1:20.
As a further improvement on the present invention, the step 3 is comprised the following steps that：
A, the ethanol water that 9-12mg lecithin, 4-6mg SDSSDC- liposomes are dissolved in 15-25mL mass concentrations 3-6%
Solution A is obtained in solution；
B, will promote bone synthetic drug be added to water or acetonitrile in be well mixed, be subsequently poured into solution A and be well mixed
To B solution, the concentration for making rush bone synthetic drug is 200-400ug/ml；
C, PLGA is dissolved in the acetonitrile solution that mass fraction is 80% C solution for obtaining that concentration is 2mg/mL, by 20-
It is well mixed in 30mL C solutions addition B solution and obtains solution D；
D, by solution D Ultrasonic Cell Disruptor ultrasound 5-10min, period is by completion of dropping 5-15mL C solutions；
Mixed liquor after the ultrasound that e, collection step d are obtained obtains final product load and promotees in centrifugal ultrafiltration three times in the super filter tube of 10kDa
Bone synthetic drug nano particle fat-polymer.
6th, a kind of preparation method for carrying rush bone synthetic drug nano particle fat-polymer according to claim 5, its
It is characterised by：To sucrose, mannose, the human blood protein or bright that addition 5-10mL concentration in solution A is 40-80mg/mL in step a
A kind of protective agent in glue solution.
As a further improvement on the present invention, the lecithin is soybean lecithin, hydrogenated soy phosphatidyl choline, yolk lecithin
One or more in fat and hydrogenated yolk lecithin.
As a further improvement on the present invention, the frequency of Ultrasonic Cell Disruptor is 21-25kHz in the step d, and power is
130-140W。
As a further improvement on the present invention, glycolide in the Poly(D,L-lactide-co-glycolide PLGA：Lactide=
20:60~85:15, the molecular weight of the Poly(D,L-lactide-co-glycolide is 2000-50000.
As a further improvement on the present invention, the bone synthetic drug that promotees is in Teriparatide or parathormone
Kind.
The sclerotin ore deposit in the bon e formation area that the bone information area and Gegenbaur's cell that the present invention is gathered using osteoclast in BMU gather
Change difference, design a kind of nano target medicine transmission system of specific selectivity, with the liposome of external SDSSDC, PLA-
Co-glycolic acid and lecithin are raw material, and add promotion bone synthetic drug, and synthesis is carried and promotees bone synthetic drug nano particle
Fat-polymer, the nano particle for loading rush bone synthetic drug has special affinity with the low hydroxyapatite of mineralization degree, enters
The bon e formation area that distribution of specific is assembled in Gegenbaur's cell after bone tissue, can greatly reduce rush bone synthetic drug and promote bone information
Side effect.Nano medicament carrying system has the advantages such as itself long circulating, targeting, slow controlled release, can overcome existing rush bone synthetic drug
The defect such as bioavailability is low, stability is poor, pharmacological action time is short.
Brief description of the drawings
Fig. 1 is the preparation process schematic diagram that load prepared by the present invention promotees bone synthetic drug nano particle fat-polymer.
Specific embodiment
The present invention is described in further detail below in conjunction with embodiment.
Polypeptide Ser-Asp-Ser-Ser-Asp-Cys abbreviations SDSSDC；
PEG-DSPE-maleimide abbreviation DSPE-PEG-Maleimide；
PEG-DSPE-succinimide abbreviation DSPE-PEG-NHS；
PEG-DSPE-silane abbreviation DSPE-PEG-Silane；
PEG-DSPE abbreviation mPEG-DSPE；
PEG-DSPE-amino abbreviation DSPE-PEG-NH2；
Poly(D,L-lactide-co-glycolide abbreviation PLGA.
Embodiment 1
It is a kind of to carry the preparation method for promoting bone synthetic drug nano particle fat-polymer：
Step one, using polypeptide solid-state reaction method prepare SDSSDC.
Step 2, the external SDSSDC on DSPE-PEG-NHS, obtain DSS6-DSPE-PEG-NHS；
The SDSSDC of 10mg is dissolved in 30mL phosphate buffer solutions first, is subsequently adding 4.8mg1- ethyl -3- (3-
Dimethyl aminopropyl)-carbodiimide and the hydroxysuccinimide-activated SDSSDC c-terminuses 3h of 3.5mgN-, add 160mg
DSPE-PEG-NHS, the hybrid reaction 18h under the gentle agitation of room, dialysis are freezed, stand-by.
Step 3, with Teriparatide, Poly(D,L-lactide-co-glycolide, hydrogenated soy phosphatidyl choline and DSS6-DSPE-
PEG-NHS is that raw material prepares load rush bone synthetic drug nano particle fat-polymer：
Glycolide in the Poly(D,L-lactide-co-glycolide：Lactide=10：3, the poly lactic-co-glycolic acid is total to
The molecular weight of polymers is 20000.
A, the second that 10mg hydrogenated soy phosphatidyl cholines, 5.5mg DSS6-DSPE-PEG-NHS are dissolved in 18mL mass concentrations 4%
Solution A is obtained in alcohol solution；Add human blood protein's aqueous solution that 6mL concentration is 50mg/mL；
B, Teriparatide is added in the acetonitrile of 30mL and be well mixed, be subsequently poured into solution A and well mixed obtain B
Solution, the concentration for making Teriparatide in B solution is 200ug/ml；
C, PLGA is dissolved in the acetonitrile solution that mass fraction is 80% C solution for obtaining that concentration is 200mg/mL, will
It is well mixed in 25mL C solutions addition B solution and obtains solution D；
D, by solution D Ultrasonic Cell Disruptor ultrasound 5min, period is by completion of dropping 8mL C solutions；The frequency of Ultrasonic Cell Disruptor
Rate is 22kHz, and power is 130W.
Mixed liquor after the ultrasound that e, collection step d are obtained obtains final product load special in centrifugal ultrafiltration three times in the super filter tube of 10kDa
Vertical handkerchief peptide nanoparticles fat-polymer.
Embodiment 2
It is a kind of to carry the preparation method for promoting bone synthetic drug nano particle fat-polymer：
Step one, using polypeptide solid-state reaction method prepare SDSSDC.
Step 2, the external SDSSDC on DSPE-PEG-NH2, obtain DSS6-DSPE-PEG-NH2；
The SDSSDC of 10mg is dissolved in 30mL phosphate buffer solutions first, is subsequently adding 5.5mg1- ethyl -3- (3-
Dimethyl aminopropyl)-carbodiimide and the hydroxysuccinimide-activated SDSSDC c-terminuses 4h of 4.3mgN-, add 180mg
DSPE-PEG-NHS, the hybrid reaction 23h under the gentle agitation of room, dialysis are freezed, stand-by.
Step 3, with Teriparatide, Poly(D,L-lactide-co-glycolide, soybean lecithin and DSS6-DSPE-PEG-NHS
Promote bone synthetic drug nano particle fat-polymer for raw material prepares to carry：
Glycolide in the Poly(D,L-lactide-co-glycolide：Lactide=13：3, the poly lactic-co-glycolic acid is total to
The molecular weight of polymers is 30000.
A, the ethanol that 9.5mg soybean lecithins, 11.5mgDSS6-DSPE-PEG-NHS are dissolved in 25mL mass concentrations 3%
Solution A is obtained in the aqueous solution；Add the aqueous sucrose solution that 8mL concentration is 50mg/ml；
B, Teriparatide is added in the acetonitrile of 50mL and be well mixed, be subsequently poured into solution A and well mixed obtain B
Solution, the concentration for making Teriparatide in B solution is 400ug/ml；
C, PLGA is dissolved in the acetonitrile solution that mass fraction is 80% C solution for obtaining that concentration is 200mg/mL, will
It is well mixed in 30mL C solutions addition B solution and obtains solution D；
D, by solution D Ultrasonic Cell Disruptor ultrasound 10min, period is by completion of dropping 15mL C solutions；Ultrasonic Cell Disruptor
Frequency is 214kHz, and power is 135W.
Mixed liquor after the ultrasound that e, collection step d are obtained obtains final product load special in centrifugal ultrafiltration three times in the super filter tube of 10kDa
Vertical handkerchief peptide nanoparticles fat-polymer.
Embodiment 3
It is a kind of to carry the preparation method for promoting bone synthetic drug nano particle fat-polymer：
Step one, using polypeptide solid-state reaction method prepare SDSSDC.
Step 2, the external SDSSDC on mPEG-DSPE, obtain DSS6-mPEG-DSPE；
The SDSSDC of 12mg is dissolved in 35mL phosphate buffer solutions first, is subsequently adding 4.8mg1- ethyl -3- (3-
Dimethyl aminopropyl)-carbodiimide and the hydroxysuccinimide-activated SDSSDC c-terminuses 3h of 3.6mgN-, add 190mg
MPEG-DSPE, the hybrid reaction 24h under the gentle agitation of room, dialysis are freezed, stand-by.
Step 3, with parathormone, Poly(D,L-lactide-co-glycolide, hydrogenated yolk lecithin and DSS6-mPEG-
DSPE is that raw material prepares load rush bone synthetic drug nano particle fat-polymer：
Glycolide in the Poly(D,L-lactide-co-glycolide：Lactide=5：1, the poly lactic-co-glycolic acid copolymerization
The molecular weight of thing is 30000.
A, the ethanol that 10mg hydrogenated yolk lecithins, 6mgDSS6-mPEG-DSPE are dissolved in into 24mL mass concentrations 6% are water-soluble
Solution A is obtained in liquid；Add human blood protein's aqueous solution that 10mL concentration is 60mg/ml；
B, parathormone is added in the water of 35mL and be well mixed, be subsequently poured into solution A and well mixed obtain B
Solution, the concentration for making Teriparatide in B solution is 250ug/ml；
C, PLGA is dissolved in the acetonitrile solution that mass fraction is 80% C solution for obtaining that concentration is 200mg/mL, will
It is well mixed in 25mL C solutions addition B solution and obtains solution D；
D, by solution D Ultrasonic Cell Disruptor ultrasound 5min, period is by completion of dropping 8mL C solutions；The frequency of Ultrasonic Cell Disruptor
Rate is 21kHz, and power is 140W.
Mixed liquor after the ultrasound that e, collection step d are obtained obtains final product load first in centrifugal ultrafiltration three times in the super filter tube of 10kDa
Parathyrine nano particle fat-polymer by shape.
Embodiment 4
It is a kind of to carry the preparation method for promoting bone synthetic drug nano particle fat-polymer：
Step one, using polypeptide solid-state reaction method prepare SDSSDC.
Step 2, the external SDSSDC on DSPE-PEG-Maleimide, obtain DSS6-DSPE-PEG-Maleimide；
The SDSSDC of 10mg is dissolved in 25mL phosphate buffer solutions first, is subsequently adding 3.6mg1- ethyl -3- (3-
Dimethyl aminopropyl)-carbodiimide and the hydroxysuccinimide-activated SDSSDC c-terminuses 4h of 2.1mgN-, add
180mgDSPE-PEG-Maleimide, the hybrid reaction 20h under the gentle agitation of room, dialysis are freezed, stand-by.
Step 3, with parathormone, Poly(D,L-lactide-co-glycolide, egg yolk lecithin and DSS6-DSPE-PEG-
Maleimide is that raw material prepares load rush bone synthetic drug nano particle fat-polymer：
Glycolide in the Poly(D,L-lactide-co-glycolide：Lactide=2：1, the poly lactic-co-glycolic acid copolymerization
The molecular weight of thing is 50000.
A, 11mg egg yolk lecithins, 4.5mgDSS6-DSPE-PEG-Maleimide are dissolved in 22mL mass concentrations 3%
Solution A is obtained in ethanol water；Add the mannose aqueous solution that 10mL concentration is 80mg/ml；
B, parathormone is added in the water of 45mL and be well mixed, be subsequently poured into solution A and well mixed obtain B
Solution, the concentration for making Teriparatide in B solution is 400ug/ml；
C, PLGA is dissolved in the acetonitrile solution that mass fraction is 80% C solution for obtaining that concentration is 200mg/mL, will
It is well mixed in 25mL C solutions addition B solution and obtains solution D；
D, by solution D Ultrasonic Cell Disruptor ultrasound 5min, period is by completion of dropping 13mL C solutions；The frequency of Ultrasonic Cell Disruptor
Rate is 25kHz, and power is 130W.
Mixed liquor after the ultrasound that e, collection step d are obtained obtains final product load first in centrifugal ultrafiltration three times in the super filter tube of 10kDa
Parathyrine nano particle fat-polymer by shape.
Embodiment 5
It is a kind of to carry the preparation method for promoting bone synthetic drug nano particle fat-polymer：
Step one, using polypeptide solid-state reaction method prepare SDSSDC.
Step 2, the external SDSSDC on DSPE-PEG-Maleimide, obtain DSS6-DSPE-PEG-Maleimide：
The SDSSDC of 15mg is dissolved in 40mL phosphate buffer solutions first, is subsequently adding 4.8mg1- ethyl -3- (3-
Dimethyl aminopropyl)-carbodiimide and the hydroxysuccinimide-activated SDSSDC c-terminuses 3h of 3.3mgN-, add
160mgDSPE-PEG-Maleimide, the hybrid reaction 12h under the gentle agitation of room, dialysis are freezed, stand-by.
Step 3, with Teriparatide, Poly(D,L-lactide-co-glycolide, soybean lecithin and DSS6-DSPE-PEG-
Maleimide is that raw material prepares load rush bone synthetic drug nano particle fat-polymer：
Glycolide in the Poly(D,L-lactide-co-glycolide：Lactide=20：60, the poly lactic-co-glycolic acid is total to
The molecular weight of polymers is 50000.
A, the second that 12mg soybean lecithins, 6mgDSS6-DSPE-PEG-Maleimide are dissolved in 15mL mass concentrations 3%
Solution A is obtained in alcohol solution；Add the aqueous sucrose solution that 5mL concentration is 40mg/ml；
B, Teriparatide is added in the acetonitrile of 50mL and be well mixed, be subsequently poured into solution A and well mixed obtain B
Solution, the concentration for making Teriparatide in B solution is 250ug/ml；
C, PLGA is dissolved in the acetonitrile solution that mass fraction is 80% C solution for obtaining that concentration is 200mg/mL, will
It is well mixed in 20mL C solutions addition B solution and obtains solution D；
D, by solution D Ultrasonic Cell Disruptor ultrasound 5min, period is by completion of dropping 15mL C solutions；The frequency of Ultrasonic Cell Disruptor
Rate is 24kHz, and power is 130W.
Mixed liquor after the ultrasound that e, collection step d are obtained obtains final product load special in centrifugal ultrafiltration three times in the super filter tube of 10kDa
Vertical handkerchief peptide nanoparticles fat-polymer.
Embodiment 6
It is a kind of to carry the preparation method for promoting bone synthetic drug nano particle fat-polymer：
Step one, using polypeptide solid-state reaction method prepare SDSSDC.
Step 2, the external SDSSDC on DSPE-PEG-Silane, obtain DSS6-DSPE-PEG-Silane；
The SDSSDC of 16mg is dissolved in 45mL phosphate buffer solutions first, is subsequently adding 4.5mg1- ethyl -3- (3-
Dimethyl aminopropyl)-carbodiimide and the hydroxysuccinimide-activated SDSSDC c-terminuses 4h of 3.8mgN-, add 190mg
DSPE-PEG-Silane, the hybrid reaction 22h under the gentle agitation of room, dialysis are freezed, stand-by.
Step 3, with parathormone, Poly(D,L-lactide-co-glycolide, hydrogenated yolk lecithin and DSS6-DSPE-
PEG-Silane is that raw material prepares load rush bone synthetic drug nano particle fat-polymer：
Glycolide in the Poly(D,L-lactide-co-glycolide：Lactide=2：3, the poly lactic-co-glycolic acid copolymerization
The molecular weight of thing is 20000.
A, 10.5mg hydrogenated yolk lecithins, 5mgDSS6-DSPE-PEG-Silane are dissolved in 20mL mass concentrations 5%
Solution A is obtained in ethanol water；Add the aqueous gelatin solution that 8mL concentration is 60mg/ml；
B, parathormone is added in the water of 30mL and be well mixed, be subsequently poured into solution A and well mixed obtain B
Solution, the concentration for making Teriparatide in B solution is 350ug/ml；
C, PLGA is dissolved in the acetonitrile solution that mass fraction is 80% C solution for obtaining that concentration is 200mg/mL, will
It is well mixed in 25mL C solutions addition B solution and obtains solution D；
D, by solution D Ultrasonic Cell Disruptor ultrasound 10min, period is by completion of dropping 11mL C solutions；Ultrasonic Cell Disruptor
Frequency is 21kHz, and power is 135W.
Mixed liquor after the ultrasound that e, collection step d are obtained obtains final product load first in centrifugal ultrafiltration three times in the super filter tube of 10kDa
Parathyrine nano particle fat-polymer by shape.
Testing result：
First, physical features
The load obtained to embodiment 4 promotees bone synthetic drug nano particle fat-polymer carries out NMR and the infrared inspection of Fourier
Survey, as a result show on proton magnetic spectrum and FTIR spectrum while there is the feature of SDSSDC and DSPE-PEG-Maleimide
Peak, it was demonstrated that SDSSDC is successfully connected with DSPE-PEG-Maleimide；It is scanned with transmission electron microscope, measuring average diameter is
120nm or so, display pattern is homogeneous spherical structure.
2nd, cellular uptake ability and toxicological experiment
The load rush bone synthetic drug nano particle fat-polymer obtained using MG63 cells and embodiment 1-6 is incubated aobvious altogether
Show, cell can be very good to absorb this nano-particle.
Detected using mtt assay and have no that load rush bone synthetic drug nano particle fat-polymer that embodiment 1-6 is obtained has substantially
Toxicity, is detected without obvious haemolysis and aggegation effect using 2% rabbit blood suspension.
3rd, bone affinity
Compared with the nano-particle of not connected polypeptide, the load rush bone synthetic drug nano particle fat that embodiment 1-6 is obtained-poly-
Compound has more preferable affinity with hydroxyapatite in vitro.Show it in animal body using small animal living body phosphorimager
Also there is preferable Bone targeting ability, can be deposited in bone tissue.
4th, function of resisting osteoporosis
In vivo, grouping experiment 1, negative control (the osteoporosis animal of medicine is not used)；2nd, (sclerotin is just for positive control
Normal animal)；3rd, free Teriparatide is given；4th, to give obtain in embodiment 6 and carry Teriparatide nano particle fat-polymer.
Terminate the animal anti-osteoporosis best results of 4 groups of experiment display, bone density highest after six months.
The above is only the preferred embodiment of the present invention, and protection scope of the present invention is not limited merely to above-mentioned implementation
Example, all technical schemes belonged under thinking of the present invention belong to protection scope of the present invention.It should be pointed out that for the art
Those of ordinary skill for, some improvements and modifications without departing from the principles of the present invention, these improvements and modifications
Should be regarded as protection scope of the present invention.
Claims (10)
- It is 1. a kind of to carry the preparation method for promoting bone synthetic drug nano particle fat-polymer, it is characterised in that：Step one, using polypeptide solid-state reaction method prepare SDSSDC；Step 2, the external SDSSDC on liposome, obtain SDSSDC- liposomes；Step 3, to promote bone synthetic drug, Poly(D,L-lactide-co-glycolide PLGA, lecithin and SDSSDC- liposomes as former Material is prepared to carry and promotees bone synthetic drug nano particle fat-polymer.
- 2. a kind of load according to claim 1 promotees the preparation method of bone synthetic drug nano particle fat-polymer, its feature It is：The liposome is PEG-DSPE-maleimide amine copolymer, distearoylphosphatidyl Monoethanolamine-polyethylene glycol-succinimide copolymerization, PEG-DSPE-silane copolymer, distearyl - polyethylene glycol-amino the copolymerization of acyl phosphatidyl-ethanolamine or one kind of PEG-DSPE copolymerization.
- 3. a kind of load according to claim 2 promotees the preparation method of bone synthetic drug nano particle fat-polymer, its feature It is：Synthesize comprising the following steps that for SDSSDC- liposomes in the step 2：Activated as coupling reagent using 1- ethyls -3- (3- dimethyl aminopropyls)-carbodiimides and N-hydroxy-succinamide SDSSDC c-terminus 3-4h, add liposome, at room temperature hybrid reaction 12-24h, dialysis, freeze, stand-by.
- 4. a kind of load according to claim 3 promotees the preparation method of bone synthetic drug nano particle fat-polymer, its feature It is：The SDSSDC is 1 with the mass ratio of liposome：10-1:20.
- 5. a kind of load according to claim 4 promotees the preparation method of bone synthetic drug nano particle fat-polymer, its feature It is：The step 3 is comprised the following steps that：A, the ethanol water that 9-12mg lecithin, 4-6mg SDSSDC- liposomes are dissolved in 15-25mL mass concentrations 3-6% In obtain solution A；B, will promote to be well mixed during bone synthetic drug is added to water or acetonitrile, and be subsequently poured into solution A and well mixed to obtain B molten Liquid, the concentration for making rush bone synthetic drug is 200-400ug/ml；C, PLGA is dissolved in the acetonitrile solution that mass fraction is 80% C solution for obtaining that concentration is 2mg/mL, by 20-30mL It is well mixed in C solution addition B solution and obtains solution D；D, by solution D Ultrasonic Cell Disruptor ultrasound 5-10min, period is by completion of dropping 5-15mL C solutions；Mixed liquor after the ultrasound that e, collection step d are obtained obtains final product load and promotees bone conjunction in centrifugal ultrafiltration three times in the super filter tube of 10kDa Into medicine nano particle fat-polymer.
- 6. a kind of load according to claim 5 promotees the preparation method of bone synthetic drug nano particle fat-polymer, its feature It is：To sucrose, mannose, human blood protein or the gelatin water for adding 5-10mL concentration for 40-80mg/mL in solution A in step a A kind of protective agent in solution.
- 7. a kind of load according to claim 6 promotees the preparation method of bone synthetic drug nano particle fat-polymer, its feature It is：The lecithin is the one kind in soybean lecithin, hydrogenated soy phosphatidyl choline, egg yolk lecithin and hydrogenated yolk lecithin Or it is several.
- 8. a kind of load according to claim 7 promotees the preparation method of bone synthetic drug nano particle fat-polymer, its feature It is：The frequency of Ultrasonic Cell Disruptor is 21-25kHz in the step d, and power is 130-140W.
- 9. a kind of load according to claim 8 promotees the preparation method of bone synthetic drug nano particle fat-polymer, its feature It is：Glycolide in the Poly(D,L-lactide-co-glycolide PLGA：Lactide=20:60~85:15, the polylactic acid-glycolic The molecular weight of acetic acid copolymer is 2000-50000.
- 10. a kind of load according to claim any one of 1-9 promotees the preparation side of bone synthetic drug nano particle fat-polymer Method, it is characterised in that：The bone synthetic drug that promotees is the one kind in Teriparatide or parathormone.
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| CN116211827A (en) * | 2023-03-17 | 2023-06-06 | 浙江大学 | Teriparatide solid lipid nanoparticle and preparation method and application thereof |
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| WO2013044734A1 (en) * | 2011-09-28 | 2013-04-04 | 中国科学院深圳先进技术研究院 | Bone targeted liposome and preparation method therefor |
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| CN116211827A (en) * | 2023-03-17 | 2023-06-06 | 浙江大学 | Teriparatide solid lipid nanoparticle and preparation method and application thereof |
| CN116211827B (en) * | 2023-03-17 | 2024-04-05 | 浙江大学 | Teriparatide solid lipid nanoparticle and preparation method and application thereof |
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