CN105193732A - Sirolimus sustained-release nano-microspheres and preparation method thereof - Google Patents
Sirolimus sustained-release nano-microspheres and preparation method thereof Download PDFInfo
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- CN105193732A CN105193732A CN201410245734.6A CN201410245734A CN105193732A CN 105193732 A CN105193732 A CN 105193732A CN 201410245734 A CN201410245734 A CN 201410245734A CN 105193732 A CN105193732 A CN 105193732A
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Abstract
The invention discloses sirolimus sustained-release nano-microspheres. A drug carrier of the sirolimus sustained-release nano-microspheres is linear-dendritic polymer methoxypolyethylene glycol-dendritic-poly L-lactide. The sustained release nano-microspheres are obtained in a self-assembly preparation mode. The maximum drug carrying capacity of the nano-microspheres can reach 45%, and the encapsulation rate is above 75%; besides, the nano-microspheres have the sustained release and sustained release targeting functions, have the in-vivo long-acting sustained release characteristic and can be applied to local parts for effectively reducing the toxic and side effects on the whole body.
Description
Technical field
The present invention relates to a kind of rapamycin nanoparticle sustained-release micro-spheres and prepare the method for rapamycin nanoparticle sustained-release micro-spheres in molecular self-assembling mode.
Background technology
Rapamycin (Rapamycin, RAPA), having another name called Sirolimus (Sirolimus) is a kind of Novel macrocyclic lactone immunosuppressant.After human body carries out organ transplantation, may immunologic rejection be there is, and the rapamycin good anti-immunologic rejection medicine that is exactly current popular one.But its water solublity is very poor, 2.6 micrograms can only be dissolved in the water of every milliliter, clinically or commercially all limit its range of application.Rapamycin in clinical treatment of cancer, mainly with oral, intravenous injection two kinds.But these two kinds of administering modes directly can not arrive operative site, through the circulation of organ or blood, cause can not reaching concentration needed for treatment in its concentration of lesion.So in the application of reality, need oral or inject a large amount of rapamycin drug, this likely can cause drug intoxication, brings huge drug side effect, also adds financial burden to patient simultaneously.
Polymer drug-carried Nano microsphere has good penetrance, slow-releasing and targeting, is the most promising current newtype drug induction system.And dendrimer, dendritic polymer has non-immunogenicity due to it, the immunoreation of cell can not be caused, hereditary-less toxicity and cytotoxicity is low, nano level particle diameter makes it easier through the biological barrier such as blood vessel wall or cell membrane, having in molecular structure can the huge cavity of packaging medicine molecule, drug loading is high, instability or insoluble drug can be wrapped up, increase its stability or improve its dissolubility and bioavailability, and Drug controlled release, molecular structure outer surface has a large amount of functional groups, its circulation time in vivo can be increased after suitable modification, obvious advantages such as realizing target administration also can be connected with targeting group, wide development and application space is had in pharmacy and pharmaceutics.
Summary of the invention
The object of the invention is a kind of rapamycin nanoparticle sustained-release micro-spheres, it adopts the mode of self assembly to prepare, the most high drug load of this Nano microsphere can reach more than 45%, and there is slow release, slow release target function, there is long-acting slow-release feature in vivo, local can be applied to and effectively reduce the toxic and side effects of whole body.
A first aspect of the present invention is a kind of rapamycin nanoparticle sustained-release micro-spheres, and its medicine carrying body is linear-dendritic macromole poly glycol monomethyl ether-dendroid-poly (l-lactic acid).
Preferably, the poly glycol monomethyl ether-dendroid-poly (l-lactic acid) of medicine carrying body described in sustained-release micro-spheres of the present invention is by mPEG
z-G
x-PLLA
yrepresent, wherein mPEG represents linear backbone poly glycol monomethyl ether, Z represents the relative molecular weight of linear backbone, G in Gx represents and props up by dihydromethyl propionic acid the dendritic structure being combined to and obtaining, X is expressed as the composition algebra of tree, PLLA represents the polylactic acid side chain of grafting, and Y represents the relative molecular weight of each polylactic acid side chain, and wherein X is 1 ~ 6; Y is 5000 ~ 10000; Z is 350 ~ 20000.
Preferably, its mean diameter of sustained-release micro-spheres of the present invention is 100 ~ 1000nm, and drug loading is 1 ~ 45%.
Preferably, in the structure of the poly glycol monomethyl ether-dendroid-poly (l-lactic acid) of medicine carrying body described in sustained-release micro-spheres of the present invention, X is 1 ~ 6, relative molecular weight Z be 5000 ~ 10000 mPEG linear backbone be connected to 2
xindividual PLLA grafting segment, the relative molecular weight Y of each segment is 2500 ~ 10000.When X is 4, namely 4 generation poly glycol monomethyl ether-dendroid-poly (l-lactic acid) chemical structural formula be:
Wherein R represents PLLA, and n is the chain number of polyoxyethylene ether on mPEG main chain, and its value makes the relative molecular weight of mPEG main chain be 5000 ~ 10000.
A second aspect of the present invention is a kind of method preparing rapamycin nanoparticle sustained-release micro-spheres of the present invention, wherein said sustained-release micro-spheres is formed by dialysis self assembly, comprising: rapamycin and poly glycol monomethyl ether-dendroid-poly (l-lactic acid) are dissolved in interior phase solvent as oil phase by (1); (2) oil phase is put into outer phase solvent and carry out dialysis treatment; (3) dialysis is complete to interior phase solvent volatilization, obtains polypeptide drug-loaded micelle solution; And (4) polypeptide drug-loaded micelle solution is isolated to carrier micelle precipitate.
The present invention also provides a kind of method preparing claim rapamycin nanoparticle sustained-release micro-spheres of the present invention, wherein said sustained-release micro-spheres is formed by solvent evaporation method self assembly, comprising: rapamycin and poly glycol monomethyl ether-dendroid-poly (l-lactic acid) are dissolved in interior phase solvent as oil phase by (1); (2) be added dropwise to by oil phase in outer phase solvent, when opalescence appears in solution, polymer self assembles forms carrier micelle; (3) phase solvent in reduction vaporization removing, obtains polypeptide drug-loaded micelle solution; And (4) polypeptide drug-loaded micelle solution obtains carrier micelle precipitate through centrifugalize.
In preparation method of the present invention, preferably, in oil phase, the mass ratio of rapamycin and poly glycol monomethyl ether-dendroid-poly (l-lactic acid) is 10 ~ 100:100; Described interior phase solvent is selected from one or more in oxolane, acetone, methanol, dichloromethane and chloroform; And described outer phase solvent is selected from one or more in water, methanol, ethanol, propanol and isopropyl alcohol.
In preparation method of the present invention, preferably, in step (2), dialysis or dropwise operation carry out in temperature 10 ~ 40 DEG C, pH6 ~ 8 time, more preferably carry out at temperature 15 ~ 35 DEG C.
In preparation method of the present invention, the carrier micelle precipitate obtained is made obtain rapamycin medicament-carried nano sustained-release micro-spheres dry powder through washing, lyophilization.
The present invention for different administering modes, can prepare the rapamycin nanoparticle sustained-release micro-spheres of different-grain diameter size, different drug loading and different slow release speed.Adopt dialysis and mixed solvent volatility process to prepare the rapamycin nanoparticle microsphere that various mean diameter is 100nm ~ 1000nm, drug loading is 1% ~ 45%, the entrapment efficiency in preparation is 75 ~ 95%.The rapamycin nanoparticle medicine carrying microballoons prepared, has the features such as long-acting slow-release, targeting and good medicine stability in vivo, can apply and local and effectively reduce the toxic and side effects of function whole body.
Accompanying drawing explanation
Fig. 1 is mPEG
5000-G
4nmr spectrum before and after-OH protection.
Fig. 2 is mPEG
5000-G
4-PLLA
5000with mPEG
5000the nmr spectrum contrast of-G4-OH.
Fig. 3 is the SEM figure of the rapamycin nanoparticle medicine carrying microballoons that embodiment 1 obtains, and wherein polymeric material adopts mPEG
5000-G
1-PLLA
5000.
Fig. 4 is the SEM figure of the rapamycin nanoparticle medicine carrying microballoons that embodiment 1 obtains, and wherein polymeric material adopts mPEG
5000-G
3-PLLA
5000.
Fig. 5 is the SEM figure of the rapamycin nanoparticle medicine carrying microballoons that embodiment 1 obtains, and wherein polymeric material adopts mPEG
5000-G
4-PLLA
5000.
Fig. 6 is the SEM figure of the rapamycin nanoparticle medicine carrying microballoons that embodiment 1 obtains, and wherein polymeric material adopts mPEG
5000-G
2-PLLA
10000.
Fig. 7 is respectively the elution profiles of the poly glycol monomethyl ether-dendroid-poly (l-lactic acid) rapamycin nanoparticle sustained-release micro-spheres of the different structure form according to embodiments of the invention 1.
Detailed description of the invention
A kind of rapamycin nanoparticle sustained-release micro-spheres of the present invention, its medicine carrying body is linear-dendritic macromole poly glycol monomethyl ether-dendroid-poly (l-lactic acid).
Poly glycol monomethyl ether-dendroid-poly (l-lactic acid) used, its general formula of molecular structure can be expressed as mPEG
z-G
x-PLLA
yrepresent, wherein G represents the dendritic structure obtained by dihydromethyl propionic acid branching, and X is the composition algebra (branching) of dendrimer, dendritic polymer, and Y represents the relative molecular weight Mn of polylactic acid side chain, and Z is the relative molecular weight Mn of poly glycol monomethyl ether; Composition algebra X=1 ~ 6; Molecular weight Y=5000 ~ 10000 of polylactic acid side chain; Z=350 ~ 20000.
Such as, the mPEG of X=4, Z ≈ 5000 is connected to 16 PLLA grafting segments, and the molecular weight Y=5000 of each segment R, its general structure can be expressed as mPEG
5000-G
4-PLLA
5000, its chemical structural formula is:
N in above-mentioned chemical structural formula is the chain number of polyoxyethylene ether, and in the structure above that its value is, the relative molecular weight of mPEG backbone portion is about 5000, and its numerical value is about (Z-32.04)/44.05.
The present invention prepares the method for rapamycin nanoparticle sustained-release micro-spheres, dialysis self assembly can be selected to form nano controlled-release microsphere, comprising: be dissolved in interior phase solvent rapamycin and poly glycol monomethyl ether-dendroid-poly (l-lactic acid) as oil phase; Oil phase is put into outer phase solvent and carry out dialysis treatment; Dialysis is complete to interior phase solvent volatilization, obtains polypeptide drug-loaded micelle solution; And polypeptide drug-loaded micelle solution is isolated to carrier micelle precipitate.
Or the present invention prepares the method for rapamycin nanoparticle sustained-release micro-spheres, dripping method self assembly can be selected to form nano controlled-release microsphere, comprising: rapamycin and poly glycol monomethyl ether-dendroid-poly (l-lactic acid) are dissolved in interior phase solvent as oil phase; Be added dropwise to by oil phase in outer phase solvent, when opalescence appears in solution, polymer self assembles forms carrier micelle; Phase solvent in reduction vaporization removing, obtains polypeptide drug-loaded micelle solution; And polypeptide drug-loaded micelle solution obtains carrier micelle precipitate through centrifugalize.
Phase solvent in as above, one or more preferably in oxolane, acetone, methanol, dichloromethane and chloroform.Outer phase solvent as above, one or more preferably in water, methanol, ethanol, propanol and isopropyl alcohol.
Separation micellar solution described in dialysis, preferably centrifugalize, such as, for high speed centrifugation is separated, centrifuge speed 8000 ~ 16000r/min, the time is 10 ~ 20min.Precipitate obtains sustained-release micro-spheres through washing and lyophilization (temperature is preferably-45 ~-20 DEG C).
In dialysis, preferably, outer phase solvent is changed in timing.Only relevant with the replacing number of times of outer phase solvent for " timing is changed ", and the replacing of outer phase solvent, evaporation rate just in order to accelerate interior phase solvent, thus shorten dialysis time to improve preparation efficiency, for performance not any impact of final rapamycin nanoparticle sustained-release micro-spheres, so replacement frequency can be grasped voluntarily by those skilled in the art.
The number of times of washing can be grasped voluntarily by those skilled in the art, the object of washing is the medicine that removing sticks to nano controlled-release microsphere outer surface, and the number of times of washing is relevant with the purity of microsphere and drug loading, and washing times is more, the purity of microsphere is higher, but drug loading is lower simultaneously.Experimental result shows, and preferably, washing times is advisable with 1 ~ 3 time.
Polylactic acid, polyethers and ester group contained in linearly of the present invention-dendritic macromole poly glycol monomethyl ether-dendroid-poly (l-lactic acid) molecule are all biodegradable; And, its structure is controlled, can by regulating the chain length of polyethers, the chain length of polylactic acid chain segment and the composition algebra in synthesis, obtain the polymer samples of different structure and performance, owing to containing hydrophilic and hydrophobic segment in molecular structure simultaneously, at suitable solvent under condition, this polymer energy self assembly formed compound with regular structure, uniform particle diameter, in have the polymer micelle of cavity.Therefore, using this polymer as the pharmaceutical carrier of rapamycin, the drug loading of medicine can not only be improved, and can according to the requirement of application, the control of being synthesized by polymer architecture, obtains the carried medicine sustained-release microsphere of different-grain diameter and sustained releasing character.In addition, the present invention adopts the mode of self assembly to prepare rapamycin nanoparticle sustained-release micro-spheres, and the microsphere obtained has that particle diameter is less, neat appearance, simple operation and other advantages, and these are the keys ensureing pharmacy quality.The polymer of different structure form (the different polylactic acid chain molecular weight of different PEG molecular weight, different algebraical sum) is adopted in rapamycin medicament-carried nano sustained-release micro-spheres preparation method of the present invention, obtained rapamycin medicament-carried nano sustained-release micro-spheres has following feature: the mean diameter of (1) rapamycin nanoparticle sustained-release micro-spheres is 100nm ~ 1000nm, and distribution uniform; (2) drug loading of rapamycin nanoparticle sustained-release micro-spheres reaches as high as more than 45%; (3) rapamycin nanoparticle sustained-release micro-spheres has good sustained release performance, and adjusts its slow-release time by the structure of polymer.
Explanation about the self assembly of preparation method Middle molecule: the system that can realize self assembly, except the feature of material structure own, also needs extraneous certain condition.Generally can be summarized as:
Molecule is amphiphilic structure.So-called amphiphilic structure, to refer in molecule simultaneously containing hydrophilic segment and oleophylic segment, as surfactant, block copolymer etc. as hyper-dispersant.Suitable solvent pair, comprises two solvents here, and one is can the solvent (in the preparation of carrier micelle or microsphere, also requiring must simultaneously dissolve used medicine) of dissolve polymer; Another is referred to as " selective solvent ", and it can mix with the segment of in parents' molecule, and immiscible with another segment, simultaneously also must be compatible and immiscible with medicine with another solvent complete (or in the concentration range of testing).Like this, when dripping in the solvent being dissolved with parents' molecule and medicine or infiltrating " selective solvent ", the solubility property of whole solvent (being made up of solvent and " selective solvent ") there occurs change, the dissolubility of parents' molecule and medicine is deteriorated, this is that it will obtain best system stability, the segment mixed with " selective solvent " will be tending towards surface, the segment immiscible with " selective solvent " and medicine will be tending towards inner, and form the aggregation of certain form.This process is constantly adding or infiltrate and spontaneously to carry out along with " selective solvent ", in addition, also needs the conditions such as the temperature be suitable for.Self assembly for molecule adopts the method for adding " selective solvent ", and " selective solvent " mode of adding is generally dialysis and dripping method.Dialysis operation controls simple, product favorable reproducibility, and the product quality that dripping method obtains is relevant with mixing speed to dropping, product quality is difficult to control relatively, the Nano microsphere performance prepared is stable not, so the present invention preferably adopts dialysis to carry out the self assembly of molecule.In addition, in the preparation process of rapamycin nanoparticle sustained-release micro-spheres, the control for temperature and pH value: temperature is lower, although be more conducive to the stable of medicine, but unfavorable to micelle self assembly, it all may cannot form stable, complete micelle in preparation in several days; And pH does not affect medicine, but polylactic acid is had to the effect promoting its degraded, polymeric material is decomposed, polylactic acid is degraded, and rate of release can be made to accelerate, and is unfavorable for application.Therefore, preparation temperature with 15 ~ 35 DEG C, pH is advisable with neutrality.
In order to understand the present invention better, below in conjunction with accompanying drawing, several embodiments of the present invention are described.
the preparation of carrier polymer
Poly glycol monomethyl ether-dendroid-poly (l-lactic acid) is prepared as required by experimenter.
Preparation method:
(1) dihydromethyl propionic acid 2,2-dimethoxypropane is carried out hydroxyl protection.
(2) with dehydrant, the dihydromethyl propionic acid of hydroxyl protection is passed through the higher anhydride of dehydrating condensation synthesizing activity.
(3) adopt divergent method to be added to by anhydride on the poly glycol monomethyl ether that molecular weight is Z by branching repeatedly, synthesize the dendrimer initiator obtaining different algebraically, its general formula of molecular structure is expressed as mPEG
z-G
x-OH, wherein X is the composition algebra of dendrimer, dendritic polymer.
(4) be grafted on dendrimer initiator by L-lactide by ring-opening polymerisation, obtained poly glycol monomethyl ether-dendroid-poly (l-lactic acid), its general formula of molecular structure can be expressed as mPEG
z-G
x-PLLA
yrepresent, wherein X is the composition algebra of dendrimer, dendritic polymer, and Y represents the relative molecular weight of poly (l-lactic acid); Composition algebra X=1 ~ 6; Molecular weight Y=5000 ~ 10000 of poly (l-lactic acid) chain; Molecular weight Z=350 ~ 20000 of poly glycol monomethyl ether.
Carrier polymer preparation example
MPEG
5000-G
4-PLLA
5000synthetic method, it includes following steps: (1) takes 50g dihydromethyl propionic acid and is dissolved in acetone, adds 2,2-dimethylpropane 69ml and 3.55g p-methyl benzenesulfonic acid.React 1-10h at normal temperatures.Add 2.5ml triethylamine, after continuing reaction a few minutes, revolve and steam except desolventizing.Add 500ml dichloromethane, sucking filtration removing insoluble matter.Dewater with anhydrous calcium chloride after solution is washed twice, vacuum drying.(2) dried product is dissolved in dichloromethane, stirring and dissolving.Dicyclohexylcarbodiimide solution is added in product slowly.24h is reacted under room temperature.After filtering reacting liquid, revolve and steam except desolventizing.Add 500ml n-hexane dissolution, after sucking filtration removes insoluble matter, recrystallization obtains anhydride.(3) initiator (mPEG is taken
5000or the initiator in each generation) 1mmol and DMAP 2.75mmol in distilled dichloromethane, add anhydride equimolar with hydroxyl in initiator, reaction 24h.Add methanol 5ml, continue reaction 24h.After question response terminates, with ice ether, product is precipitated out.Dried in vacuo overnight.The product of drying is dissolved in methanol, adds acidic cationic resin, stirring reaction 24h.Ice ether sedimentation is dry, obtains product mPEG
5000-G
1-OH.Repeat step (3), obtain mPEG
5000-G
2-OH.This step of repeatable operation is until obtain mPEG
5000-G
4-OH dendrimer initiator.(4) 0.5gmPEG is taken
5000-G
4-OH is in polymerization bottle.Add 8.0gL-lactide, using stannous octoate as catalyst under anhydrous and oxygen-free environment sealing by fusing polymerization bottle, reactant is polymerized 18 hours at 80 ~ 130 DEG C.After being dissolved by product, by absolute ether precipitation, filter, in vacuum drying oven after dry 24h, the micro-yellow powder shape solid obtained is product mPEG
5000-G
4-PLLA
5000.
Molecular structure characterization:
By methods such as nuclear-magnetisms, the polymer that preparation example obtains is characterized, to prove the molecular structure of product.
Fig. 1 is mPEG
5000-G
4nmr spectrum before and after-OH protection.After hydroxyl protection is mPEG
5000-G
4: δ 3.38 (s, 3H), δ 3.45 ~ 3.85 (m, 452H), δ 4.20 ~ 4.40 (m, 16H), δ 1.23 ~ 1.30 (m, 21H), δ 4.14 (t, 3H), δ 1.14 (s, 24H), δ 1.32 (s, 24H), δ 1.35 (s, 24H).Be mPEG after deprotection
5000-G
4-OH: δ 3.38 (s, 3H), δ 3.45 ~ 3.85 (m, 452H), δ 4.20 ~ 4.40 (m, 16H), δ 1.22 ~ 1.38 (m, 21H), δ 1.08 (s, 24H).Do not have other obviously assorted peaks, between each peak, the ratio of peak area is substantially equal with H atom number ratio, illustrates that the product formula of this molecular structural formula and experimental design matches.Contrast mPEG
5000-G
4and mPEG
5000-G
4the proton nmr spectra graph discovery of-OH, the methyl peak (l, m) be originally positioned near 1.32ppm and 1.35ppm disappears completely after deprotection, and this illustrates mPEG
5000-G
4the success of dendroid deprotection, obtains product mPEG
5000-G
4-OH.
Fig. 2 is mPEG
5000-G
4-PLLA
5000with mPEG
5000-G
4the nmr spectrum contrast of-OH.The characteristic peak of poly (l-lactic acid) can be observed significantly, i.e. two stack features peaks near 5.25ppm and 1.75ppm from the nuclear-magnetism figure of copolymer.And according to integral and calculating, the structure of this polymer is mPEG
5000-G
4-PLLA
5000.
Use and the similar method of above-mentioned preparation example, by the dosage of L-lactide in the number of repetition of rate-determining steps (3) and step (4), mPEG used in embodiment below can be obtained respectively
5000-G
1-PLLA
5000, mPEG
5000-G
3-PLLA
5000, and mPEG
10000-G
2-PLLA
10000, mPEG
5000-G
1-PLLA
2500, mPEG
5000-G
3-PLLA
2500.
The present invention tests rapamycin used and is provided by pharmaceutical factory, and specification meets States Pharmacopoeia specifications.
From current result of study, the dialysis kind of material and the size of molecular cut off, appreciable impact is not had on the performance of medicament-carrying nano-microsphere, so bag filter can according to those skilled in the art need select, and bag filter material used in the present invention is regenerated cellulose (RC), molecular cut off is 2000 ~ 7000.In embodiment, high speed centrifugation is separated, centrifuge speed 8000 ~ 16000r/min, and the time is 10 ~ 20min; Cryodesiccated temperature-65 ~-20 DEG C.
The mensuration of rapamycin nanoparticle sustained-release micro-spheres (hereinafter referred to as microcapsule) drug loading and envelop rate:
Take the medicine carrying microballoons dry powder of 3mg, after it being dissolved completely with the dichloromethane of 10ml, adopt ultraviolet spectrophotometer under 278nm, measure the absorbance of solution, calculated the content of rapamycin by standard curve.And calculate drug loading and envelop rate.
Gross mass × 100% of the amount/medicine carrying microballoons of drug loading=medicine carrying microballoons Chinese medicine;
Amount/the experiment of envelop rate=medicine carrying microballoons Chinese medicine adds amount × 100% of medicine.
Embodiment 1
A preparation method for rapamycin medicament-carried nano sustained-release micro-spheres, it includes following steps: (1) takes four kinds of structures and is respectively mPEG
5000-G
1-PLLA
5000, mPEG
5000-G
3-PLLA
5000, mPEG
5000-G
4-PLLA
5000and mPEG
5000-G
2-PLLA
10000poly glycol monomethyl ether-dendroid-each 5.0mg of poly (l-lactic acid) polymer, after being dissolved in the oxolane of 8mL respectively, respectively add the rapamycin of 4.0mg, as oil phase after ultrasonic 15min makes rapamycin dissolve completely under the ultrasound condition of 400W; (2) four oil phases being transferred to molecular cut off is respectively in the bag filter of 7000, bag filter is placed in respectively the beaker filling 60ml deionization redistilled water, the water bath with thermostatic control that beaker is placed in 35 DEG C is dialysed, and the external solution in 8 hours replacing beakers; (3) when dialysis time reaches 24 hours, treat that solvents tetrahydrofurane volatilization is complete, in bag filter, obtain polypeptide drug-loaded micelle solution; (4) polypeptide drug-loaded micelle solution prepared by the poly glycol monomethyl ether-dendroid-PLLA polymer of four different structures is separated through high speed centrifugation respectively, and with 15mL ultrafiltration water washing three times, centrifugalize obtains white powder precipitate, the rapamycin medicament-carrying nano-microsphere that mean diameter is respectively 230,300.2,311.5 and 328.4nm is obtained after lyophilization, the coefficient of dispersion of its particle size distribution is respectively 0.247,0.181,0.165 and 0.185, and drug loading is respectively 42.1%, 41.5%, 33.5% and 38.5%.
Embodiment 2
A preparation method for rapamycin medicament-carried nano sustained-release micro-spheres, it includes following steps: (1) takes three kinds of structures and is respectively mPEG
5000-G
1-PLLA
2500, mPEG
5000-G
3-PLLA
2500and mPEG
5000-G
4-PLLA
2500poly glycol monomethyl ether-dendroid-each 5.0mg of poly (l-lactic acid) polymer, after being dissolved in the oxolane of 8mL respectively, respectively add the rapamycin of 3.0mg, as oil phase after ultrasonic 15min makes rapamycin dissolve completely under the ultrasound condition of 400W; (2) three oil phases being transferred to molecular cut off is respectively in the bag filter of 7000, bag filter is placed in respectively the beaker filling 60ml deionization redistilled water, the water bath with thermostatic control that beaker is placed in 35 DEG C is dialysed, and the external solution in 4 hours replacing beakers; (3) when dialysis time reaches 24 hours, treat that solvents tetrahydrofurane volatilization is complete, in bag filter, obtain polypeptide drug-loaded micelle solution; (4) polypeptide drug-loaded micelle solution prepared by the poly glycol monomethyl ether-dendroid-poly (l-lactic acid) polymer of three different structures is separated through high speed centrifugation respectively, and with 15mL ultrafiltration water washing three times, centrifugalize obtains white powder precipitate, the rapamycin medicament-carrying nano-microsphere that mean diameter is respectively 211.1,281.9 and 296.7nm is obtained after lyophilization, the coefficient of dispersion of its particle size distribution is respectively 0.213,0.155 and 0.134, and drug loading is respectively 30.7%, 32.6% and 33.29%.
Embodiment 3
A preparation method for rapamycin medicament-carried nano sustained-release micro-spheres, it includes following steps: molecular structure is mPEG by (1)
5000-G
4-PLLA
5000poly glycol monomethyl ether-dendroid-poly (l-lactic acid) polymer 5.0mg be dissolved in oxolane 8mL formed oil phase, the rapamycin taking 5.0mg, 4.0mg, 3.0mg and 1.0mg is respectively placed in bag filter; (2) bag filter is placed in the beaker of the deionization redistilled water of 60ml, the water bath with thermostatic control that beaker is placed in 35 DEG C carries out dialysis 24 hours, treats that solvents tetrahydrofurane diffusion is complete, obtains polypeptide drug-loaded micelle solution in bag filter; Centrifugalize obtains white powder precipitate, obtain mean diameter after lyophilization and be respectively 343.7nm, 311.5nm, 287.4nm and 347.0nm, breadth coefficient is respectively the rapamycin medicament-carrying nano-microsphere of 0.290,0.165,0.185 and 0.231, its drug loading is respectively 47.8%, 42.1%, 34.8% and 13.9%, and envelop rate is respectively 95.4%, 94.7%, 93.0% and 83.6%.
Embodiment 4
A preparation method for rapamycin medicament-carried nano sustained-release micro-spheres, it includes following steps: molecular structure is mPEG by (1)
5000-G
4-PLLA
2500poly glycol monomethyl ether-dendroid-poly (l-lactic acid) polymer 5.0mg be dissolved in oxolane 8mL formed oil phase, the rapamycin taking 5.0mg, 4.0mg, 3.0mg and 1.0mg is respectively placed in bag filter; (2) bag filter is placed in the beaker of the deionization redistilled water of 60ml, the water bath with thermostatic control that beaker is placed in 35 DEG C carries out dialysis 24 hours, treats that solvents tetrahydrofurane diffusion is complete, obtains polypeptide drug-loaded micelle solution in bag filter; Centrifugalize obtains white powder precipitate, obtain mean diameter after lyophilization and be respectively 316.5nm, 282.1nm, 249.6nm and 300.4nm, breadth coefficient is respectively the rapamycin medicament-carrying nano-microsphere of 0.238,0.216,0.134 and 0.316, its drug loading is respectively 44.6%, 40.6%, 33.3% and 10.3%, and envelop rate is respectively 89.2%, 91.4%, 88.9% and 82.4%.
Embodiment 5
A preparation method for rapamycin medicament-carried nano sustained-release micro-spheres, it includes following steps: (1) takes molecular structure is mPEG
5000-G
4-PLLA
5000poly glycol monomethyl ether-dendroid-poly (l-lactic acid) polymer 5.0mg, after being dissolved in the acetone of 8mL, add the rapamycin of 4.0mg, as oil phase after ultrasonic 15min makes rapamycin dissolve completely under the ultrasound condition of 400W; (2) oil phase being transferred to molecular cut off is in the bag filter of 7000, bag filter is placed in the beaker filling 60ml deionization redistilled water, and the water bath with thermostatic control that beaker is placed in 35 DEG C is dialysed, and the external solution in 8 hours replacing beakers; (3) when dialysis time reaches 24 hours, treat that solvent acetone volatilization is complete, in bag filter, obtain polypeptide drug-loaded micelle solution; Obtain the rapamycin medicament-carrying nano-microsphere that mean diameter is 198.0nm after lyophilization, its particle size distribution coefficient is 0.116, and drug loading is 35.7%, and envelop rate is 95.1%.
Embodiment 6
A preparation method for rapamycin medicament-carried nano sustained-release micro-spheres, it includes following steps: (1) takes molecular structure is mPEG
5000-G
4-PLLA
5000poly glycol monomethyl ether-dendroid-each 5.0mg of two parts, poly (l-lactic acid) polymer, after being dissolved in the acetone of 8mL respectively, respectively add the rapamycin of 4.0mg, as oil phase after ultrasonic 15min makes rapamycin dissolve completely under the ultrasound condition of 400W; (2) two oil phases being transferred to molecular cut off is respectively in the bag filter of 7000, bag filter is placed in respectively the beaker filling 60ml deionization redistilled water, the water bath with thermostatic control that beaker is placed in 30 DEG C, 25 DEG C is respectively dialysed, and the external solution in 8 hours replacing beakers; (3) when dialysis time reaches 24 hours, treat that solvent acetone volatilization is complete, in bag filter, obtain polypeptide drug-loaded micelle solution; Obtain the rapamycin medicament-carrying nano-microsphere that mean diameter is respectively 212.1nm, 235.4nm after lyophilization, its particle size distribution coefficient is 0.148,0.176, and drug loading is respectively 33.4%, 30.8%, and envelop rate is 89.2%, 82.3%.
Embodiment 7
A preparation method for rapamycin medicament-carried nano sustained-release micro-spheres, it includes following steps: (1) takes structure is respectively mPEG
5000-G
4-PLLA
5000and mPEG
5000-G
4-PLLA
2500after poly glycol monomethyl ether-dendroid-each 3mg of poly (l-lactic acid) polymer is dissolved in the oxolane of 5mL respectively, add 2.5mg rapamycin respectively, as oil phase after 200W makes rapamycin dissolve completely in ultrasonic 15 minutes in a water bath; (2) 25 DEG C, under the magnetic agitation of 600r/min, the organic solution of 5mL is dropped in the deionized water of 10ml respectively with the speed of 0.05ml/ minute, obtains respective polypeptide drug-loaded micelle solution; (3) solution is transferred in the water bath with thermostatic control of 33 DEG C the 8h that dialyses, removing oxolane organic solvent; (4) obtain the nano drug-carrying microsphere that mean diameter is respectively 85.1nm, 73.2nm after lyophilization, its particle size distribution coefficient is 0.187,0.174, and drug loading is respectively 40.0% and 37.50%, and envelop rate is respectively 76.9% and 75.4%.
Embodiment 8
A preparation method for rapamycin medicament-carried nano sustained-release micro-spheres, it includes following steps: (1) takes molecular structure is respectively mPEG
5000-G
4-PLLA
5000poly glycol monomethyl ether-dendroid-each 5.0mg of three parts, poly (l-lactic acid) polymer, be dissolved in respectively oxolane 8mL formed oil phase, the rapamycin respectively taking 4.0mg is placed in bag filter; (2) respectively bag filter is placed in the beaker of the deionization redistilled water of 60ml, the water bath with thermostatic control that beaker is placed in 25 DEG C, 30 DEG C and 35 DEG C respectively carries out dialysis 24 hours, treats that solvents tetrahydrofurane diffusion is complete, obtains polypeptide drug-loaded micelle solution in bag filter; Centrifugalize obtains white powder precipitate, obtain mean diameter after lyophilization and be respectively 250.2,273.5 and 311.5nm, breadth coefficient is respectively the rapamycin medicament-carrying nano-microsphere of 0.153,0.198 and 0.165, its drug loading is respectively 26.9%, 32.0% and 42.1%, and envelop rate is respectively 60.6%, 71.9% and 94.7%.
The particle size distribution of the rapamycin medicament-carrying nano-microsphere of the various embodiments described above adopts the ZetasizerNano laser particle analyzer of Malvern company of Britain to detect.
According to embodiment, the feature of the rapamycin nanoparticle sustained-release micro-spheres prepared by the present invention can be provided:
(1) particle diameter is less and be evenly distributed, and particle diameter, between 100 ~ 1000nm, all belongs to the scope of nano drug-carrying microsphere, therefore can pass through blood transport.And the medicine carrying microballoons of different-grain diameter can be obtained by the polymer of different molecular structures;
(2) drug loading is high, envelop rate is high, and in the ordinary course of things, drug loading is between 30% ~ 40%, and envelop rate is between 80% ~ 95%;
(3) preparation owing to adopting dialysis and solvent evaporation method to carry out self assembly, therefore, as long as the condition of the relatively good controls such as the rate of charge of control temperature and polymer and medicine, the medicine carrying microballoons that repeatability is fabulous can be obtained, comparatively other such as method such as emulsifying or emulsion, the regularity, repeatability etc. of micelle are more stable, as shown in accompanying drawing 3 ~ Fig. 6, can ensure that medical effect etc. is stablized.
(4) the release in vitro time is long, 10 ~ 40 days can be reached, as shown in Figure 7, for the elution profiles of the rapamycin nanoparticle sustained-release micro-spheres of three kinds of materials of 1 preparation according to the preferred embodiment, rapamycin nanoparticle sustained-release micro-spheres initial release speed can be found out, in 10 hours, the Cumulative release amount of medicine is between 10% to 50%, reason mainly due to two aspects causes burst drug release: one is the quick release of the medicine being adsorbed on microsphere surface, two is inner due to microsphere large with the drug level difference of outer aqueous phase, make the diffusion velocity of the rapamycin of microsphere inside than very fast.
But after being greater than 1 day release time, along with the raising of outer aqueous phase drug level, the drug releasing rate of medicine carrying microballoons eases up gradually.Reach 200 hours upon release (about 17 days), the cumulative release amount of three kinds of microspheres is also respectively between 40% to 90%.Meanwhile, because the structure of three kinds of polymer is different, along with the increase of algebraically, the amount of polylactic acid increases, and makes microsphere finer and close, the drug diffusion rate in medicine carrying microballoons is slowed down, cause different rate of release curves.Therefore, the molecular structure of medicine carrier polymer can be designed according to the demand of clinical medicine rate of release.
Claims (10)
1. a rapamycin nanoparticle sustained-release micro-spheres, is characterized in that, medicine carrying body is linear-dendritic macromole poly glycol monomethyl ether-dendroid-poly (l-lactic acid).
2. sustained-release micro-spheres as claimed in claim 1, wherein said medicine carrying body poly glycol monomethyl ether-dendroid-poly (l-lactic acid) is by mPEG
z-G
x-PLLA
yrepresent, wherein mPEG represents linear backbone poly glycol monomethyl ether, Z represents the relative molecular weight of linear backbone, G in Gx represents and props up by dihydromethyl propionic acid the dendritic structure being combined to and obtaining, X is expressed as the composition algebra of tree, PLLA represents the polylactic acid side chain of grafting, and Y represents the relative molecular weight of each polylactic acid side chain, and wherein X is 1 ~ 6; Y is 5000 ~ 10000; Z is 350 ~ 20000.
3. sustained-release micro-spheres as claimed in claim 1 or 2, its mean diameter is 100 ~ 1000nm, and drug loading is 1 ~ 45%, and envelop rate is more than 75%.
4. sustained-release micro-spheres as claimed in claim 2, in the structure of wherein said medicine carrying body poly glycol monomethyl ether-dendroid-poly (l-lactic acid), X is 1 ~ 6, relative molecular weight Z be 5000 ~ 10000 mPEG linear backbone be connected to 2
xindividual PLLA grafting segment, the relative molecular weight Y of each segment is 2500 ~ 10000, when composition algebra X is 4, namely 4 generation poly glycol monomethyl ether-dendroid-poly (l-lactic acid) chemical structural formula be:
Wherein R represents PLLA, and n is the chain number of polyoxyethylene ether on mPEG main chain, and its value makes the relative molecular weight of mPEG main chain be 5000 ~ 10000.
5. prepare a method for the rapamycin nanoparticle sustained-release micro-spheres described in any one of claim 1-4, wherein said sustained-release micro-spheres is formed by dialysis self assembly, comprising:
(1) rapamycin and poly glycol monomethyl ether-dendroid-poly (l-lactic acid) are dissolved in interior phase solvent as oil phase;
(2) oil phase is put into outer phase solvent and carry out dialysis treatment;
(3) dialysis is complete to interior phase solvent volatilization, obtains polypeptide drug-loaded micelle solution; And
(4) polypeptide drug-loaded micelle solution is isolated to carrier micelle precipitate.
6. prepare a method for the rapamycin nanoparticle sustained-release micro-spheres described in any one of claim 1-4, wherein said sustained-release micro-spheres is formed by dripping method self assembly, comprising:
(1) rapamycin and poly glycol monomethyl ether-dendroid-poly (l-lactic acid) are dissolved in interior phase solvent as oil phase;
(2) be added dropwise to by oil phase in outer phase solvent, when opalescence appears in solution, polymer self assembles forms carrier micelle;
(3) phase solvent in reduction vaporization removing, obtains polypeptide drug-loaded micelle solution; And
(4) polypeptide drug-loaded micelle solution obtains carrier micelle precipitate through centrifugalize.
7. the method as described in claim 5 or 6, in oil phase, the mass ratio of rapamycin and poly glycol monomethyl ether-dendroid-poly (l-lactic acid) is 10 ~ 100:100; Described interior phase solvent is selected from one or more in oxolane, acetone, methanol, dichloromethane and chloroform; And described outer phase solvent is selected from one or more in water, methanol, ethanol, propanol and isopropyl alcohol.
8. the method as described in claim 5 or 6, in step (2) operate in temperature 10 ~ 40 DEG C, pH6 ~ 8 time are carried out.
9. the method as described in claim 5 or 6, operates in temperature 15 ~ 35 DEG C in step (2).
10. the method as described in claim 5 or 6, carrier micelle precipitate is made obtain rapamycin medicament-carried nano sustained-release micro-spheres dry powder through washing, lyophilization.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109431997A (en) * | 2018-12-20 | 2019-03-08 | 武汉科福新药有限责任公司 | A kind of rapamycin locally injecting preparation and preparation method thereof |
| CN110623925A (en) * | 2019-09-26 | 2019-12-31 | 严鹏科 | Rapamycin nanometer sustained release agent and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102462656A (en) * | 2010-11-04 | 2012-05-23 | 天津德兰玮诚生物技术有限公司 | Macrolide immunosuppressant drug loaded nanomicelle and preparation method thereof |
| CN103242517A (en) * | 2013-05-17 | 2013-08-14 | 中国药科大学 | Preparation of multifunctional linear-dendritic segmented copolymer and application in pharmaceutics thereof |
| CN103748142A (en) * | 2011-05-13 | 2014-04-23 | 加利福尼亚大学董事会 | Reversibly crosslinked micelle systems |
-
2014
- 2014-06-05 CN CN201410245734.6A patent/CN105193732A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102462656A (en) * | 2010-11-04 | 2012-05-23 | 天津德兰玮诚生物技术有限公司 | Macrolide immunosuppressant drug loaded nanomicelle and preparation method thereof |
| CN103748142A (en) * | 2011-05-13 | 2014-04-23 | 加利福尼亚大学董事会 | Reversibly crosslinked micelle systems |
| CN103242517A (en) * | 2013-05-17 | 2013-08-14 | 中国药科大学 | Preparation of multifunctional linear-dendritic segmented copolymer and application in pharmaceutics thereof |
Non-Patent Citations (3)
| Title |
|---|
| YVONNE HED,ET AL: "Side-by-Side Comparison of Dendritic-Linear Hydrids and Their Hyperbranched Analogs as Micellar Carriers of Chemotherapeutics", 《JOURNAL OF POLYMER SCIENCE》 * |
| 江琳等: "雷帕霉素缓释胶束的制备及其释放行为", 《华东理工大学学报（自然科学版）》 * |
| 谢威等: "树枝状聚L-丙交酯-dendron-聚乙二醇-dendron-聚L-丙交酯的自组装", 《华东理工大学学报（自然科学版）》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109431997A (en) * | 2018-12-20 | 2019-03-08 | 武汉科福新药有限责任公司 | A kind of rapamycin locally injecting preparation and preparation method thereof |
| CN110623925A (en) * | 2019-09-26 | 2019-12-31 | 严鹏科 | Rapamycin nanometer sustained release agent and preparation method thereof |
| WO2021057007A1 (en) * | 2019-09-26 | 2021-04-01 | 严鹏科 | Rapamycin nanoscale sustained-release agent and preparation method thereof |
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