CN108341923A - Glycerin monostearate-(polyethylene glycol)2Copolymer and preparation method and carrier micelle - Google Patents
Glycerin monostearate-(polyethylene glycol)2Copolymer and preparation method and carrier micelle Download PDFInfo
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- CN108341923A CN108341923A CN201810178434.9A CN201810178434A CN108341923A CN 108341923 A CN108341923 A CN 108341923A CN 201810178434 A CN201810178434 A CN 201810178434A CN 108341923 A CN108341923 A CN 108341923A
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- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G18/00—Polymeric products of isocyanates or isothiocyanates
- C08G18/06—Polymeric products of isocyanates or isothiocyanates with compounds having active hydrogen
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- 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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- C08G18/06—Polymeric products of isocyanates or isothiocyanates with compounds having active hydrogen
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Abstract
The present invention provides a kind of glycerin monostearates (polyethylene glycol) with structure shown in Formulas I2Copolymer, glycerin monostearate (polyethylene glycol) provided by the invention2Copolymer is made of hydrophilic fractions and hydrophobic patches, the nano-micelle using hydrophilic fractions as shell, hydrophobic patches for kernel can be formed when self assembly in water, and drug can pass through hydrophobic interaction force effect with the hydrophobic patches, to be wrapped in the inside of the nano-micelle, this not only increases the dissolubility of drug in water, also reduce the toxic side effect of simple drug.
Description
Technical field
The present invention relates to biomedical material technology more particularly to a kind of glycerin monostearates-(polyethylene glycol)2
Copolymer and preparation method thereof and carrier micelle.
Background technology
Amphipathic nature block polymer refers to the block copolymer that those contain hydrophilic segment and hydrophobic segment, in aqueous solution
In be easy to that microphase-separated occurs, and be self-assembled into a variety of accumulation shapes, such as vesica, micella.Amphipathic nature block polymer is certainly
Control release and the load treatment base of hydrophobic drug can be realized as drug carrier system by assembling the nano-micelle formed
Cause and albumen, thus can greatly improve the bioavilability and stability of drug.Polyethylene glycol (PEG), which is one kind, to be had
The crystalline polymer material of biocompatibility, is used widely in biotechnology and field of pharmacology.Currently, most of
Amphipathic nature block polymer be all based on PEG synthesis, this analog copolymer has the characteristic of thermal sensitivity and " stealthy ".
Currently, the anticancer drug of most of Clinical practices generally has water solubility bad, cytotoxicity is tall and big, lacks treatment
Selectivity, into blood after easy-clear fail the shortcomings of, this also causes utilization ratio of drug relatively low, and improves dosage and administration
Frequency can cause higher toxic side effect again.Therefore, hydrophobic drug is contained with amphipathic pharmaceutical carrier, can be effectively changed
Many deficiencies of kind drug, and can will be in drug delivery to tumour cell.
Invention content
In view of this, the purpose of the present invention is to provide a kind of glycerin monostearate-(polyethylene glycol)2Copolymer and its
Preparation method and carrier micelle, glycerin monostearate-(polyethylene glycol) provided by the invention2Copolymer is applied to as carrier
In carrier micelle, cytotoxicity is low, and slow release effect and stability are good.
In order to achieve the above-mentioned object of the invention, the present invention provides following technical scheme：
The present invention provides a kind of glycerin monostearate-(polyethylene glycol)2Copolymer has structure shown in Formulas I：
In Formulas I, n=4~136.
Preferably, n=11~43.
The present invention provides glycerin monostearate-described in above-mentioned technical proposal (polyethylene glycol)2The preparation side of copolymer
Method includes the following steps：
Under protective atmosphere, glycerin monostearate, hexamethylene diisocyanate and organic solvent are mixed and carried out in advance
Poly- reaction, obtains prepolymer；Wherein, the molar ratio of the glycerin monostearate and hexa-methylene diisocyanate is 1：(2~
3)；
The prepolymer is mixed with poly glycol monomethyl ether, organic solvent and carries out polymerisation, is obtained with shown in Formulas I
The glycerin monostearate-(polyethylene glycol) of structure2Copolymer, wherein mole of the prepolymer and poly glycol monomethyl ether
Than being 1：(2~2.1)；
The poly glycol monomethyl ether has structure shown in Formula II：
In Formula II, n=4~136.
Preferably, the mixing of the glycerin monostearate, hexamethylene diisocyanate and organic solvent is by six Asias
Methyl diisocyanate is added drop-wise in the mixture of glycerin monostearate and organic solvent, and the rate of the dropwise addition is 12~30
Drop/minute.
Preferably, the temperature of the prepolymerization reaction is 55~65 DEG C, and the time of prepolymerization reaction is 2.5~3.5h.
Preferably, the mixing of the prepolymer and poly glycol monomethyl ether, organic solvent is to drip poly glycol monomethyl ether
It is added in the mixture of prepolymer and organic solvent, the rate of the dropwise addition is 12~30 drops/minute.
Preferably, the temperature of the polymerisation is 55~65 DEG C, and the time of polymerisation is 2.5~3.5h.
The present invention provides a kind of carrier micelle, including carrier and the drug that is coated by the carrier, the carrier is upper
State glycerin monostearate-described in technical solution (polyethylene glycol)2It is prepared by preparation method described in copolymer or above-mentioned technical proposal
Obtained glycerin monostearate-(polyethylene glycol)2Copolymer.
Preferably, the glycerin monostearate-(polyethylene glycol)2The mass ratio of copolymer and drug is 10：(2.5~
3.5)。
Preferably, the drug is hydrophobic drug.
The present invention provides a kind of glycerin monostearates-(polyethylene glycol) with structure shown in Formulas I2Copolymer, this
The glycerin monostearate-(polyethylene glycol) provided is provided2Copolymer is made of hydrophilic fractions and hydrophobic patches, in water
The nano-micelle for kernel using hydrophilic fractions as shell, hydrophobic patches can be formed when self assembly, and drug can be dredged with described
Aqueous segment is not only increased by hydrophobic interaction force effect to be wrapped in the inside of the nano-micelle in this way
The dissolubility of drug in water, also reduces the toxic side effect of simple drug.Embodiment the experimental results showed that, the present invention carries
The glycerin monostearate-(polyethylene glycol) of confession2The critical micelle concentration of copolymer is 5.08mg/L, illustrates the monostearate
Glyceride-(polyethylene glycol)2Copolymer can keep nano-micelle structure under conditions of diluter, have higher stability；With
The glycerin monostearate-(polyethylene glycol)2Copolymer is the carrier micelle of carrier, and the burst size of drug is in 16h
The burst size 32.6% of drug when 12.8%, 1 week, illustrates that sustained drug release effect is good；PEG-GMS-PEG is measured using mtt assay to exist
External cytotoxicity, PEG-GMS-PEG, almost without toxicity, illustrate that the monostearate is sweet to COS7 cells and HELA cells
Grease-(polyethylene glycol)2Copolymer has good biocompatibility.
Description of the drawings
The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.
Fig. 1 is glycerin monostearate-(polyethylene glycol)2The synthetic route chart of copolymer；
Fig. 2 is in embodiment 5 during prepolymerization reaction-NCO content with the prepolymerization reaction time variation diagram；
Fig. 3 is prepolymer and glycerin monostearate-(polyethylene glycol) in embodiment 52The infrared spectrogram of copolymer；
Fig. 4 is prepolymer and glycerin monostearate-(polyethylene glycol) in embodiment 52The nuclear magnetic spectrum of copolymer；
Fig. 5 is glycerin monostearate-(polyethylene glycol) in embodiment 52The gel permeation chromatography figure of copolymer；
Fig. 6 is glycerin monostearate, poly glycol monomethyl ether and glycerin monostearate-(poly- second two in embodiment 5
Alcohol)2The differential scanning calorimetric analysis curve of copolymer；
Fig. 7 is glycerin monostearate-(polyethylene glycol) in embodiment 62The critical micelle concentration figure of copolymer；
Fig. 8 is glycerin monostearate-(polyethylene glycol) in embodiment 62The grain size distribution of copolymer micelle and transmission
Electron microscope；
Fig. 9 is the drug release patterns figure of carrier micelle in embodiment 6；
Figure 10 is that mtt assay measures glycerin monostearate-(polyethylene glycol) in embodiment 62The cell of copolymer in vitro
Toxicity figure.
Specific implementation mode
The present invention provides a kind of glycerin monostearate-(polyethylene glycol)2Copolymer has structure shown in Formulas I：
In Formulas I, n=4~136, preferably 11~43.
The present invention provides glycerin monostearate-described in above-mentioned technical proposal (polyethylene glycol)2The preparation side of copolymer
Method includes the following steps：
Under protective atmosphere, glycerin monostearate, hexamethylene diisocyanate and organic solvent are mixed and carried out in advance
Poly- reaction, obtains prepolymer；Wherein, the molar ratio of the glycerin monostearate and hexa-methylene diisocyanate is 1：(2~
3)；
The prepolymer is mixed with poly glycol monomethyl ether, organic solvent and carries out polymerisation, is obtained with shown in Formulas I
The glycerin monostearate-(polyethylene glycol) of structure2Copolymer, wherein mole of the prepolymer and poly glycol monomethyl ether
Than being 1：(2~2.1)；
The poly glycol monomethyl ether has structure shown in Formula II：
In Formula II, n=4~136.
The present invention by glycerin monostearate (GMS), hexamethylene diisocyanate (HDI) and has under protective atmosphere
Solvent mixing carries out prepolymerization reaction, obtains prepolymer (NCO-GMS-NCO)；Wherein, the glycerin monostearate and six Asias
The molar ratio of methyl diisocyanate is 1:(2~3).In the present invention, the glycerin monostearate, hexa-methylene diisocyanate
Hexamethylene diisocyanate is preferably added drop-wise to glycerin monostearate and organic solvent by the mixing of ester and organic solvent
In mixture, the rate of the dropwise addition is preferably 12~30 drops/minute.In the present invention, the glycerin monostearate and have
Solvent is preferably performed under heating conditions when mixing, to ensure that glycerin monostearate can dissolve in organic solvent；Institute
The temperature for stating heating is preferably 55~65 DEG C.
The present invention does not have the organic solvent special restriction, can dissolve hydroxylated tung oil and hexa-methylene two is different
Cyanate makes it be smoothed out prepolymerization reaction, specific such as toluene or tetrahydrofuran；In the present invention, the organic solvent
Preferably anhydrous grade organic solvent.The present invention does not have special restriction for the protective gas type for providing the protective atmosphere,
It is specific such as nitrogen using protective gas well known to those skilled in the art.
In the present invention, the prepolymerization reaction preferably carries out under agitation；The rate of the stirring is preferably 200~
400rpm。
In the present invention, the temperature of the prepolymerization reaction is preferably 55~65 DEG C, more preferably 60 DEG C；The prepolymerization reaction
Time be preferably 2.5~3.5h, more preferably 3h.The present invention preferably determines the prepolymerization reaction by the content of measurement-NCO
Terminal.In an embodiment of the present invention, the content for specifically using di-n-butylamine titration measuring-NCO the, when-NCO's
Content stops heating when reaching theoretical value (16.32%), completes the prepolymerization reaction, obtains prepolymer.
After completing the prepolymerization reaction, the present invention preferably removes the organic solvent in resulting material, and surplus material is dried
To constant weight, prepolymer is obtained.The present invention does not have special restriction for method used by removal organic solvent, using this field
Method known to technical staff, such as rotary evaporation.In the present invention, the drying is preferably dried in vacuo；The vacuum
Dry temperature is preferably 25~35 DEG C, more preferably 30 DEG C；The vacuum drying vacuum degree is preferably -0.1Mpa；This hair
It is bright there is no special restriction for the vacuum drying time, the surplus material can be dried to constant weight.
After obtaining the prepolymer, the prepolymer is mixed progress with poly glycol monomethyl ether, organic solvent by the present invention
Polymerisation obtains the glycerin monostearate-(polyethylene glycol) with structure shown in Formulas I2Copolymer (PEG-GMS-PEG),
Wherein, the molar ratio of the prepolymer and poly glycol monomethyl ether is 1：(2~2.1).In the present invention, the polyethyleneglycol
Methyl ether has structure shown in Formula II：
In Formula II, n=4~136, preferably 11~43.
In the present invention, the mixing of the prepolymer and poly glycol monomethyl ether, organic solvent is by polyethyleneglycol first
Ether is added drop-wise in the mixture of prepolymer and organic solvent；The drop rate of the poly glycol monomethyl ether is preferably 12~30
Drop/minute.Poly glycol monomethyl ether is added drop-wise in prepolymer by the present invention, and poly glycol monomethyl ether can be made abundant with prepolymer
Reaction.
The present invention does not have the organic solvent special restriction, can dissolve prepolymer and poly glycol monomethyl ether,
It is set to be smoothed out polymerisation, it is specific such as toluene or tetrahydrofuran；In the present invention, the organic solvent is preferably nothing
Water grade organic solvent.
In the present invention, the polymerisation preferably carries out under agitation；The rate of the stirring is preferably 200~
400rpm。
In the present invention, the temperature of the polymerisation is preferably 55~65 DEG C, more preferably 60 DEG C；The polymerisation
Time be preferably 2.5~3.5h, more preferably 3h.The present invention preferably determines the polymerisation by the content of measurement-NCO
Terminal.In an embodiment of the present invention, the content for specifically using di-n-butylamine titration measuring-NCO the, when-NCO's
Content stops heating when reaching zero, completes the polymerisation, obtains the glycerin monostearate-with structure shown in Formulas I
(polyethylene glycol)2Copolymer.
After completing the polymerisation, the present invention preferably removes the organic solvent in resulting material, and surplus material is dried
To constant weight, the glycerin monostearate-(polyethylene glycol) with structure shown in Formulas I is obtained2Copolymer.The present invention has removal
Method used by solvent does not have special restriction, using method well known to those skilled in the art, such as rotary evaporation.
In the present invention, the drying is preferably dried in vacuo；The vacuum drying temperature is preferably 25~35 DEG C, more preferably 30
℃；The vacuum drying vacuum degree is preferably -0.1Mpa；The present invention does not have special limit for the vacuum drying time
It is fixed, the surplus material can be dried to constant weight.
The present invention provides a kind of carrier micelle, including carrier and the drug that is coated by the carrier, the carrier is upper
State glycerin monostearate-described in technical solution (polyethylene glycol)2It is prepared by preparation method described in copolymer or above-mentioned technical proposal
Obtained glycerin monostearate-(polyethylene glycol)2Copolymer.In the present invention, glycerin monostearate-(the poly- second two
Alcohol)2The mass ratio of copolymer and drug is preferably 10：(2.5~3.5), more preferably 10：3.In the present invention, the drug
Preferably hydrophobic drug；The present invention does not have the type of the hydrophobic drug special restriction, using art technology
Hydrophobic drug known to personnel, it is specific such as prednisone acetate, taxol, adriamycin, brufen or methotrexate (MTX).
The present invention does not have the preparation method of the carrier micelle special restriction, using known to those skilled in the art
The technical solution for preparing carrier micelle.In the present invention, the preparation method of the carrier micelle preferably includes following step
Suddenly：
By the glycerin monostearate-(polyethylene glycol) with structure shown in Formulas I2Copolymer is mixed with organic solvent, is obtained
To glycerin monostearate-(polyethylene glycol)2Copolymer solution；
By the glycerin monostearate-(polyethylene glycol)2Copolymer solution is dialysed in water, the monostearate
Glyceride-(polyethylene glycol)2Copolymer forms glycerin monostearate-(polyethylene glycol) by self assembly mechanism2Copolymer nano
Micella；
By the glycerin monostearate-(polyethylene glycol)2Copolymer nano micella, drug and organic solvent mixing, will
Gained mixed solution is dialysed in water, obtains carrier micelle.
The present invention is preferably by glycerin monostearate-(polyethylene glycol)2Copolymer is mixed with organic solvent, obtains single tristearin
Acid glyceride-(polyethylene glycol)2Copolymer solution.In the present invention, the glycerin monostearate-(polyethylene glycol)2Copolymer
The concentration of solution is preferably 180~220mg/L, more preferably 200mg/L.The present invention does not have the type of the organic solvent
Special restriction can dissolve the glycerin monostearate-(polyethylene glycol) using well known to those skilled in the art2Copolymerization
The organic solvent of object；In the present invention, the organic solvent preferably includes tetrahydrofuran, methanol, ethyl alcohol, N, N- dimethyl
One or more of formamide and dimethyl sulfoxide.
Obtain glycerin monostearate-(polyethylene glycol)2After copolymer solution, the present invention is preferably sweet by the monostearate
Grease-(polyethylene glycol)2Copolymer solution is dialysed in water, the glycerin monostearate-(polyethylene glycol)2Copolymer
Glycerin monostearate-(polyethylene glycol) is formed by self assembly mechanism2Copolymer nano micella.The present invention is for the dialysis
There is no special restriction, the glycerin monostearate-(polyethylene glycol) can be removed2Organic solvent is in copolymer solution
It can.In the present invention, the molecular cut off of bag filter used by the dialysis is preferably 1000Da；The time of the dialysis is excellent
It is selected as 22~26h, more preferably for 24 hours；Preferably every 3.5~4.5h changes a water, more preferably 4h in the dialysis procedure.At this
In invention, the water is preferably distilled water；Carry out used water and the glycerin monostearate-(poly- second when the dialysis
Glycol)2The volume ratio of copolymer solution is preferably (180~220)：1, more preferably 200：1.
In the present invention, the glycerin monostearate-(polyethylene glycol)2Copolymer nano micella is with hydrophilic fractions
(poly glycol monomethyl ether segment) is shell, hydrophobic patches (prepolymer segment) are kernel.In the present invention, described single stearic
Acid glyceride-(polyethylene glycol)2The granularity of copolymer nano micella is preferably 50~100nm, more preferably 70~90nm.
Obtain glycerin monostearate-(polyethylene glycol)2After copolymer nano micella, the present invention is sweet by the monostearate
Grease-(polyethylene glycol)2Copolymer nano micella, drug and organic solvent mixing, gained mixed solution are carried out in water thoroughly
Analysis, obtains carrier micelle.In the present invention, the organic solvent is preferably tetrahydrofuran (THF) and n,N-Dimethylformamide
(DMF) mixed solvent；The volume ratio of the tetrahydrofuran and N,N-dimethylformamide is preferably (8~10)：1, more preferably
It is 9：1.In the present invention, the glycerin monostearate-(polyethylene glycol)2Copolymer nano micella, drug quality and have
The volume ratio of solvent is preferably (18~22) mg：(7~9) mg：(18~22) mL, more preferably 20mg：8mg：20mL.At this
In invention, the molecular cut off of bag filter used by the dialysis is preferably 1000Da；The time of the dialysis is preferably 22
~26h, more preferably for 24 hours；Preferably every 3.5~4.5h changes a water, more preferably 4h in the dialysis procedure.In the present invention
In, when the glycerin monostearate-(polyethylene glycol)2The mixed solution of copolymer nano micella, drug and organic solvent by
It is transparent when becoming translucent, it was demonstrated that the carrier micelle has been formed.
In the present invention, the glycerin monostearate-(polyethylene glycol) being made of hydrophilic fractions and hydrophobic patches2Altogether
Polymers in water self assembly when can form the nano-micelle using hydrophilic fractions as shell, hydrophobic patches for kernel, and drug
Can be with the hydrophobic patches by hydrophobic interaction force effect, to be wrapped in the inside of the nano-micelle, this
Sample not only increases the dissolubility of drug in water, also reduces the toxic side effect of simple drug.
Below in conjunction with the embodiment in the present invention, the technical solution in the present invention is clearly and completely described.It is aobvious
So, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based on the reality in the present invention
Example is applied, every other embodiment obtained by those of ordinary skill in the art without making creative efforts all belongs to
In the scope of protection of the invention.
Fig. 1 is glycerin monostearate-(polyethylene glycol)2The synthetic route of copolymer, by glycerin monostearate (GMS)
With hexamethylene diisocyanate (HDI) carry out prepolymerization reaction, obtain prepolymer (NCO-GMS-NCO), then with polyethylene glycol
Monomethyl ether (mPEG) carries out polymerisation, obtains glycerin monostearate-(polyethylene glycol)2Copolymer (PEG-GMS-PEG).
Embodiment 1
Under nitrogen protection, the dry toluene of 3.58g glycerin monostearates (GMS, 0.01mol) and 20mL are mixed,
4.24g hexamethylene diisocyanates are added dropwise with the rate of 12 drops/minute into gained mixture in the stirring and dissolving at 60 DEG C
(HDI, 0.0252mol), carries out prepolymerization reaction 3h under 60 DEG C, stirring condition after being added dropwise ,-NCO content reaches theoretical value
(16.32%), revolving removes the dry toluene in resulting material, is dried under vacuum to constant weight under the conditions of 30 DEG C, -0.1Mpa, obtains
To prepolymer (NCO-GMS-NCO)；
Under nitrogen protection, the prepolymer is mixed with 20mL dry toluenes, into gained mixture with 12 drops/minute
Rate be added dropwise 11g poly glycol monomethyl ethers (mPEG, 0.02mol, Mn=550), react 3h at 60 DEG C ,-NCO content reaches
Zero, revolving removes the dry toluene in resulting material, is dried under vacuum to constant weight under the conditions of 30 DEG C, -0.1Mpa, obtains single tristearin
Acid glyceride-(polyethylene glycol)2Copolymer (PEG-GMS-PEG550).
Embodiment 2
Glycerin monostearate-(polyethylene glycol) is prepared according to the method for embodiment 12Copolymer (PEG-GMS-
PEG750), wherein the molecular weight Mn=750 of poly glycol monomethyl ether.
Embodiment 3
Glycerin monostearate-(polyethylene glycol) is prepared according to the method for embodiment 12Copolymer (PEG-GMS-
PEG1000), wherein the molecular weight Mn=1000 of poly glycol monomethyl ether.
Embodiment 4
Glycerin monostearate-(polyethylene glycol) is prepared according to the method for embodiment 12Copolymer (PEG-GMS-
PEG1900), wherein the molecular weight Mn=1900 of poly glycol monomethyl ether.
Embodiment 5
The prepolymer and glycerin monostearate-(polyethylene glycol) prepared to embodiment 12Copolymer is analyzed, specifically
It is as follows：
Fig. 2 be prepolymerization reaction during-NCO content with the prepolymerization reaction time variation diagram, as shown in Figure 2, when pre-polymerization is anti-
When being 3h between seasonable ,-NCO content meets with theoretical value substantially；If continuing to extend the reaction time at this time, not only increase energy
Consumption, and the side reaction between HDI can be caused, cause-NCO content decline.Therefore, 3h is the most suitable prepolymerization reaction time.
Fig. 3 is prepolymer and glycerin monostearate-(polyethylene glycol)2The infrared spectrogram of copolymer, wherein (A)
NCO-GMS-NCO, (B) PEG-GMS-PEG.3344cm in Fig. 3-1For the stretching vibration absworption peak of the N-H of hydrogen bonding, 2924cm-1
And 2859cm-1The asymmetry and symmetrical stretching vibration absorption peak of respectively C-H, 1723cm-1It is stretched for C=O in carbamate
Contracting vibration absorption peak, 1532cm-1II bands of a spectrum of urethano amide to be formed are coupled for N-H in-plane bending vibrations and C-N stretching vibrations
Absorption peak, 1248cm-1The III band absorption peak of urethano amide to be formed is coupled for N-H in-plane bending vibrations and C-N stretching vibrations,
1110cm-1For the stretching vibration absworption peak of ehter bond.NCO-GMS-NCO is in 2263cm-1There is one strong peak (peaks-NCO) in place,
And there are urethane bond and without hydroxyl, showing HDI to react with the hydroxyl in GMS, to generate terminal isocyanate group pre-
Polymers；PEG-GMS-PEG is in 2263cm-1The vibration absorption peak of place-NCO disappears substantially, shows that-the NCO in reaction system is basic
The reaction was complete, can tentatively conclude and synthesize target product PEG-GMS-PEG.
Fig. 4 is prepolymer and glycerin monostearate-(polyethylene glycol)2The nuclear magnetic spectrum of copolymer, wherein (a) (NCO-
GMS-NCO), (b) (PEG-GMS-PEG).The ownership at each peak is as follows in Fig. 4：Belong to methene proton peak d+i, the e+h on HDI,
F+g is respectively at 3.13ppm, 1.47ppm, 1.24ppm；Peak o at δ 3.54ppm belongs to the proton peak of mPEG methylenes；δ
Peak p at 3.36ppm belongs to the proton peak of methyl in mPEG；Peak n at δ 0.86ppm belongs to the proton in GMS terminal methyls, δ
Peak a at 4.88ppm, 4.18ppm, b are respectively belonging to the proton in the methine and methylene that are connected with ester group in GMS, δ
Peak j at 2.29ppm belongs to the proton in the methylene being connected with carbonyl in GMS, and the peak m at δ 1.31ppm belongs to GMS
In proton in the methylene that is connected with terminal methyl, the peak l at δ 1.24ppm, 3.63ppm, k belong to remaining methylene in GMS
In proton δ 1.24ppm at peak.Pass through1H-NMR collection of illustrative plates is it can be seen that all kinds of protons of target product PEG-GMS-PEG
Characteristic peak, but never see the proton peak c of H above amide group N, this is because the H above N disappears because proton exchange acts on
It loses, it is difficult to be observed in spectrogram.Infrared spectrum and nuclear magnetic resonance hydrogen spectruming determining are the result shows that present invention success synthesizing amphipathic
Glycerin monostearate-(polyethylene glycol)2Copolymer.
Fig. 5 is glycerin monostearate-(polyethylene glycol)2The gel permeation chromatography figure of copolymer, as can be seen from Figure 5
PEG-GMS-PEG is in Unimodal Distribution in gel permeation chromatography (GPC) test, and breadth coefficient Mw/Mn is 1.66, illustrates product group
At single, and the number-average molecular weight measured is 2116, is closer to the molecular weight 1795 of theoretical calculation.
Fig. 6 is glycerin monostearate, poly glycol monomethyl ether and glycerin monostearate-(polyethylene glycol)2Copolymer
Differential scanning calorimetric analysis curve, heating rate be 10 DEG C/min.The fusing point of GMS and mPEG is respectively as can be seen from Figure 6
66 DEG C and 18 DEG C, and there are dual melting peaks in PEG-GMS-PEG, are located at 12 DEG C and 30 DEG C.Compared to pure GMS and
MPEG, fusing point decrease, and show to obtain is PEG-GMS-PEG triblock copolymers, rather than the mixing of GMS and mPEG
Object.
Embodiment 6
Glycerin monostearate-(polyethylene glycol) prepared by embodiment 12Copolymer is mixed with tetrahydrofuran respectively, is obtained
To glycerin monostearate-(polyethylene glycol)2Copolymer solution；
By the glycerin monostearate-(polyethylene glycol)2It is the saturating of 1000Da that copolymer solution, which goes to molecular cut off,
It analyses in bag, dialyses in 2L distilled water for 24 hours, first water is changed to remove tetrahydrofuran, the glycerin monostearate-per 4h
(polyethylene glycol)2Copolymer forms glycerin monostearate-(polyethylene glycol) by self assembly mechanism2Copolymer nano micella；
By the glycerin monostearate-(polyethylene glycol)2Copolymer nano micella 20mg, prednisone acetate 8mg and
20mL organic solvents (mixed solvent of 18mL THF and 2mLDMF) mix, and gained mixed solution, which is gone to molecular cut off, is
It dialyses in the bag filter of 1000Da and (changes first water per 4h) for 24 hours, obtain carrier micelle.
The carrying drug ratio of the carrier micelle is 12.57%, encapsulation rate 32.21%.
Fig. 7 is glycerin monostearate-(polyethylene glycol)2The critical micelle concentration figure of copolymer.Amphipathic nature polyalcohol exists
The minimum concentration that micella is self-assembly of in water is critical micelle concentration, is less than this concentration, polymer is in water with single molecular chain
Form exist, and higher than this concentration when, polymer, which can be assembled, is self-assembly of micella, thus we using pyrene be fluorescence probe survey
Determine the critical micelle concentration (CMC) of polymer micelle, the results are shown in Figure 7.When PEG-GMS-PEG micelle forma-tions, pyrene can divide
Cloth is in the hydrophobic core of micella, and the local environment around pyrene becomes nonpolarity from polarity, due to this change, in excitation spectrum
In will appear red shift (by 336 to 340nm).By the way that the peak height ratios at 340nm and 336nm are mapped with micellar solution concentration,
It was found that in the low concentration range, I340/I336 is varied less；After concentration reaches certain value, I340/I336 continues to increase, table
It is bright to initially form PEG-GMS-PEG micellas.Therefore concentration is the CMC value of PEG-GMS-PEG at the inflection point of low concentration region in Fig. 7.
As can be seen from Figure 7 the critical micelle concentration of PEG-GMS-PEG is 5.08mg/L, shows PEG-GMS-PEG in extremely dilute ring
Nano-micelle structure can be kept in border, and there is higher stability.
Fig. 8 is glycerin monostearate-(polyethylene glycol)2The grain size distribution and transmission electron microscope picture of copolymer micelle.Scheme A
Show that the average grain diameter of PEG-GMS-PEG micellas is 80.7nm, the coefficient of dispersion 0.373.Scheme B and shows PEG-GMS-PEG micellas
Grain size be about 30nm, be distributed in uniform spherical.The two grain size the difference is that, A survey be PEG-GMS-PEG micellas water
Grain size is closed, what B was surveyed is the grain size under PEG-GMS-PEG micella dry states.
Fig. 9 be carrier micelle at 37 DEG C, drug under the conditions of phosphate buffer (PBS, 0.1mol/L, pH=7.4)
Release conditions.The acetic acid when burst size of prednisone acetate is 12.8%, 1 week in carrier micelle in 16h it can be seen from Fig. 9
The burst size of Bo Nisong is 32.6%, is in sustainable lasting releasing trend.
Figure 10 is that mtt assay measures glycerin monostearate-(polyethylene glycol)2The cytotoxicity figure of copolymer in vitro,
In, (A) COS7 cells, (B) HELA cells.It can be seen from fig. 10 that in surveyed concentration range, PEG-GMS-PEG is to just
The survival rate of normal COS7 cells and HELA cells is almost 100%, it was demonstrated that PEG-GMS-PEG illustrates PEG- almost without toxicity
GMS-PEG is the good high molecular material of a kind of biocompatibility, is had laid a good foundation as pharmaceutical carrier for it.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of glycerin monostearate-(polyethylene glycol)2Copolymer has structure shown in Formulas I：
In Formulas I, n=4~136.
2. glycerin monostearate-(polyethylene glycol) according to claim 12Copolymer, which is characterized in that the n=11
~43.
3. glycerin monostearate-described in claims 1 or 2 (polyethylene glycol)2The preparation method of copolymer, includes the following steps：
Under protective atmosphere, it is anti-that glycerin monostearate, hexamethylene diisocyanate and organic solvent are mixed into progress pre-polymerization
It answers, obtains prepolymer；Wherein, the molar ratio of the glycerin monostearate and hexa-methylene diisocyanate is 1：(2~3)；
The prepolymer is mixed with poly glycol monomethyl ether, organic solvent and carries out polymerisation, is obtained with structure shown in Formulas I
Glycerin monostearate-(polyethylene glycol)2Copolymer, wherein the molar ratio of the prepolymer and poly glycol monomethyl ether is
1：(2~2.1)；
The poly glycol monomethyl ether has structure shown in Formula II：
In Formula II, n=4~136.
4. preparation method according to claim 3, which is characterized in that the glycerin monostearate, hexa-methylene two are different
The mixing of cyanate and organic solvent is that hexamethylene diisocyanate is added drop-wise to glycerin monostearate and organic solvent
In mixture, the rate of the dropwise addition is 12~30 drops/minute.
5. preparation method according to claim 3, which is characterized in that the temperature of the prepolymerization reaction is 55~65 DEG C, in advance
The time of poly- reaction is 2.5~3.5h.
6. preparation method according to claim 3, which is characterized in that the prepolymer and poly glycol monomethyl ether, organic
The mixing of solvent is added drop-wise to poly glycol monomethyl ether in the mixture of prepolymer and organic solvent, and the rate of the dropwise addition is
12~30 drops/minute.
7. preparation method according to claim 3, which is characterized in that the temperature of the polymerisation is 55~65 DEG C, is gathered
The time for closing reaction is 2.5~3.5h.
8. a kind of carrier micelle, including carrier and the drug that is coated by the carrier, the carrier is described in claims 1 or 2
Glycerin monostearate-(polyethylene glycol)2The list that any one of copolymer or claim 3~7 preparation method are prepared
Tristerin-(polyethylene glycol)2Copolymer.
9. carrier micelle according to claim 8, which is characterized in that the glycerin monostearate-(polyethylene glycol)2Altogether
The mass ratio of polymers and drug is 10：(2.5~3.5).
10. carrier micelle according to claim 8 or claim 9, which is characterized in that the drug is hydrophobic drug.
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| CN101831067A (en) * | 2010-05-31 | 2010-09-15 | 王二新 | Polyethylene glycol ester conjugate and application thereof in medicine preparation |
| CN102337073A (en) * | 2011-08-18 | 2012-02-01 | 中科院广州化学有限公司 | Water-soluble nonluminous anionic electrophoretic coating and preparation method and application thereof |
| CN102391498A (en) * | 2011-08-17 | 2012-03-28 | 沈阳药科大学 | Polyethylene glycol di-fatty acid glyceride derivatives and application thereof to medicine delivery |
| CN104774309A (en) * | 2015-04-16 | 2015-07-15 | 武汉科技大学 | Star amphipathic polyurethane and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101831067A (en) * | 2010-05-31 | 2010-09-15 | 王二新 | Polyethylene glycol ester conjugate and application thereof in medicine preparation |
| CN102391498A (en) * | 2011-08-17 | 2012-03-28 | 沈阳药科大学 | Polyethylene glycol di-fatty acid glyceride derivatives and application thereof to medicine delivery |
| CN102337073A (en) * | 2011-08-18 | 2012-02-01 | 中科院广州化学有限公司 | Water-soluble nonluminous anionic electrophoretic coating and preparation method and application thereof |
| CN104774309A (en) * | 2015-04-16 | 2015-07-15 | 武汉科技大学 | Star amphipathic polyurethane and preparation method thereof |
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