CN110358063A - The preparation method of block copolymer mPEG-b-PLA - Google Patents
The preparation method of block copolymer mPEG-b-PLA Download PDFInfo
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- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G63/00—Macromolecular compounds obtained by reactions forming a carboxylic ester link in the main chain of the macromolecule
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Abstract
The invention discloses the preparation method of block copolymer mPEG-b-PLA, preparation method is the following steps are included: pre-process raw material poly glycol monomethyl ether and lactide；Poly glycol monomethyl ether and lactide are dissolved respectively again, amidine class catalyst is added in poly glycol monomethyl ether, is added with stirring lactide, 10-180min is polymerize under oxygen free condition, reaction is then terminated using terminator, obtains crude product；After crude product is dissolved with eluant, eluent, then pass through the isolated purified product of chromatographic column.The present invention provides a kind of high yield of amphipathic nature block polymer, high-purity, the preparation methods of Narrow Molecular Weight Distribution.The preparation method is polymerisation in solution, and organic catalysis can accurately control reaction condition；It can be purified respectively using chromatography and centrifugal process.The molecular weight dispersity of the block copolymer of acquisition can control within 1.1.
Description
Technical field
The invention belongs to field of pharmaceutical chemistry technology, it is related to synthesis and the purification process of a kind of block copolymer, especially relates to
And a kind of preparation method of block copolymer mPEG-b-PLA.
Background technique
Block copolymer mPEG-b-PLA is the good carrier of curative drug, and it is for example anticancer can to increase insoluble medicine
The solubility of medicine reduces side effects of pharmaceutical drugs etc..Since the synthetic reaction of the block polymer is ring-opening polymerisation, the prior art is normal
It is bulk polymerization with method, but bulk polymerization needs high temperature, the reaction time is usually also required to 12h or more, and polymerizing condition is difficult to surely
Fixed control, and production efficiency is low, and the distribution of molecular weight of product and molecular weight is difficult to control.
Summary of the invention
The technical problem to be solved in the present invention is that in view of the drawbacks of the prior art, provide anti-under a kind of high efficiency, room temperature
It answers, the reaction time is short, the preparation method of the di-block copolymer mPEG-b-PLA of the product of enough acquisition Narrow Molecular Weight Distributions.
The technical solution adopted by the present invention to solve the technical problems is:
A kind of preparation method of block copolymer mPEG-b-PLA, comprising the following steps:
(1) pretreatment of raw material: using poly glycol monomethyl ether and lactide as raw material, poly glycol monomethyl ether is dissolved in organic
After solvent, remove water, filter it is spare；By lactide recrystallization, drying for standby；
(2) synthesis of mPEG-b-PLA: poly glycol monomethyl ether and lactide are weighed, wherein poly glycol monomethyl ether: third
The weight ratio of lactide is (0.1~30): 1；Poly glycol monomethyl ether and lactide are dissolved with volatile organic solvent respectively
To poly glycol monomethyl ether solution and Lactide solution；Then amidine class catalyst is added in poly glycol monomethyl ether, then is stirring
Lower addition lactide is mixed, polymerize 10-180min under oxygen free condition, reaction is then terminated using terminator, obtains crude product；Its
In, amidine class catalyst loading is the 0.1%~3% of reaction raw materials total mole number；The additive amount of terminator is that reaction raw materials are total
The 0.1%~3% of molal quantity；
(3) column chromatographic purifying: after crude product is dissolved with eluant, eluent, then passing through the isolated purified product of chromatographic column,
Middle eluant, eluent is volatile organic solvent.The volatile organic solvent of preferred steps (3) be low carbon chain alcohol or alkane substitute or
The mixture of both persons.
Further, it in the preparation method of the block copolymer mPEG-b-PLA, preferably after step (3), also wraps
Include following steps:
(4) o/w/ prepares micella: the purified product that column chromatography for separation is crossed is dissolved in volatile organic solvent, preferably low carbon chain
Alkanes dissolution after mix with water, mixed with water, it is fully emulsified, stir volatilize completely to organic solvent, obtain micella；
(5) it is centrifuged: micella being subjected to centrifugal treating, takes supernatant that end product is lyophilized to obtain.
Further, in the preparation method of the block copolymer mPEG-b-PLA, the preferably described volatility is organic molten
Agent selects alkane, alkane substitute or the alcohol of low carbon chain, and the carbon chain lengths are C1-C10。
Further, in the preparation method of the block copolymer mPEG-b-PLA, in the preferably described step (2), nothing
Oxygen condition is under vacuum or in inert gas.
Further, in the preparation method of the block copolymer mPEG-b-PLA, in the preferably described step (2), institute
Stating terminator is organic acid.It is preferred that at least one of benzoic acid, glacial acetic acid, ethanedioic acid, succinic acid.
Further, in the preparation method of the block copolymer mPEG-b-PLA, the preferably described amidine class catalyst is
TBD, DBN or DBU.
Further, in the preparation method of the block copolymer mPEG-b-PLA, in the preferably described step (3), institute
The filler for stating column chromatography is sephadex, and granularity is 100~400 mesh.
Further, in the preparation method of the block copolymer mPEG-b-PLA, in the preferably described step (3), institute
The filler for stating column chromatography is the glucose gel of G type or LH type, and the filler of the preferably described column chromatography is G25, G50, G100, LH20
Or the glucose gel of LH60.
Further, in the preparation method of the block copolymer mPEG-b-PLA, in the preferably described step (3), institute
Stating eluant, eluent is methanol, ethyl alcohol, methylene chloride, chloroform or two or more of mixtures in them.
The method that polymerisation in solution is used in the present invention, by amidine class catalyst, at normal temperature in the short time
Block polymerization reaction is completed, the present invention is led to by control feed ratio, catalyst and the dosage and the reaction time that use terminator
The resultant effect for crossing above-mentioned various conditions achievees the purpose that the molecular weight and its molecular weight distribution that accurately control product.
The present invention is chromatographed with density centrifugation both methods using column to bi-block copolymer after the completion of reaction, then respectively
Product is purified.Wherein, since the molecule of the different spaces volume of column chromatography can by time different characteristic in filler
To separate the small molecular weight compounds such as monomer, catalyst, terminator after reaction in system and polymer, reach purifying
Purpose.Centrifugal method is that the particle based on different-grain diameter has the different rates of settling, and amphiphilic polymer is easy in water
Form micella, prepare micella using o/w method, amphiphilic polymer can form the micella of different-grain diameter according to molecular weight, finally from
Heart method can allow the copolymer of different molecular weight to separate, and reach purifying.
Detailed description of the invention
Present invention will be further explained below with reference to the attached drawings and examples, in attached drawing:
Fig. 1 is the nuclear magnetic resonance map of the embodiment of the present invention 1；
Fig. 2 is the nuclear magnetic resonance map of the embodiment of the present invention 2；
Fig. 3 is the gpc chromatogram of the embodiment of the present invention 2；
Fig. 4 is the nuclear magnetic resonance map of the embodiment of the present invention 3；
Fig. 5 is the nuclear magnetic resonance map of the embodiment of the present invention 4；
Fig. 6 is the gpc chromatogram of the embodiment of the present invention 4；
Fig. 7 is the gpc chromatogram after the embodiment of the present invention 4 is centrifuged；
Fig. 8 is the nuclear magnetic resonance map of the embodiment of the present invention 5；
Fig. 9 is the nuclear magnetic resonance map of the embodiment of the present invention 6；
Figure 10 is the absorbance release profiles of sustained release experiment of the embodiment of the present invention；
Figure 11 is the nuclear magnetic resonance map of comparative example；
Figure 12 is the gpc chromatogram of comparative example.
Specific embodiment
For a clearer understanding of the technical characteristics, objects and effects of the present invention, now control attached drawing is described in detail
A specific embodiment of the invention.
A kind of block copolymer mPEG-b-PLA is the high molecular polymer for including hydrophily section and hydrophobicity section, wherein
Hydrophily section is the poly glycol monomethyl ether of molecular weight 300-20000, and hydrophobicity section is the polylactic acid of molecular weight 200-15000,
Hydrophobicity section is blocked with organic acid row, and the weight ratio of hydrophily section and hydrophobicity section is (0.1~30): 1.In above-mentioned molecular weight
In range, can arbitrarily it select, such as: mPEG1000-b-PLA9000, mPEG1000-b-PLA3000, mPEG1000-b-
PLA1000、mPEG5000-b-PLA3000、mPEG5000-b-PLA5000、mPEG5000-b-PLA8500、mPEG1000-b-
PLA10000,mPEG6000-b-PLA15000,mPEG6000-b-PLA200,mPEG300-b-PLA5000.Above-mentioned block copolymerization
Object mPEG-b-PLA is prepared by following preparation method.
A kind of preparation method of block copolymer mPEG-b-PLA, comprising the following steps:
(1) feedstock processing: weighing poly glycol monomethyl ether and lactide, and it is organic that poly glycol monomethyl ether is dissolved in volatility
After solvent, remove water, filter it is spare；By lactide recrystallization, drying for standby；Wherein, poly glycol monomethyl ether: the weight of lactide
Than for (0.1~30): 1；The alkane or alkane substitute of the volatile organic solvent selection low carbon chain, the carbon chain lengths are
C1-C10。
(2) synthesis of mPEG-b-PLA: poly glycol monomethyl ether and lactide are dissolved with volatile organic solvent respectively
To poly glycol monomethyl ether solution and Lactide solution, the alkane of the preferred low carbon chain of the volatile organic solvent of this step or
Alkane substitute, the carbon chain lengths are C1-C10.Then amidine class catalyst is added in poly glycol monomethyl ether solution, then is stirring
Lower addition Lactide solution is mixed, polymerize 10-180min under oxygen free condition, reaction is then terminated using terminator, obtains crude product；
Wherein, amidine class catalyst loading is the 0.1%~3% of reaction raw materials total mole number；The terminator is organic acid, terminator
Additive amount be reaction raw materials total mole number 0.1%~3%, the terminator be preferably benzoic acid, glacial acetic acid, ethanedioic acid,
At least one of succinic acid.The amidine class catalyst is preferably TBD, DBN or DBU.Under the preferred vacuum of oxygen free condition or inertia
In gas, wherein the vacuum degree under vacuum is 0.08~0.1MPa, and inert gas selects nitrogen or argon gas.
(3) column chromatographic purifying: after crude product is dissolved with eluant, eluent, then passing through the isolated purified product of chromatographic column,
Middle eluant, eluent is volatile organic solvent, and the preferably low carbon chain alcohol of low carbon chain or alkane substitute either low carbon chain alcohol and takes
For the mixture of alkane, most preferably methanol, ethyl alcohol, methylene chloride, chloroform, tetrachloromethane or they in two or more
Mixture.The filler of the column chromatography is sephadex, and granularity is 100~400 mesh.The filler of column chromatography is preferably
The glucose gel of G type or LH type.The filler of the most preferably described column chromatography is the glucose of G25, G50, G100, LH20 or LH60
Gel.
After purification, can with the following steps are included:
(4) o/w/ prepares micella: the purified product volatile organic solvent that column chromatography for separation is crossed, preferably low carbon chain
It is mixed after alkanes dissolution with water, fully emulsified, stirring is volatilized completely to organic solvent, obtains micella；
(5) it is centrifuged: micella being subjected to centrifugal treating, takes supernatant that end product is lyophilized to obtain.
It is described in detail below with specific embodiment:
Embodiment 1, a kind of preparation method of block copolymer mPEG-b-PLA (mPEG1000-b-PLA9000), including with
Lower step:
(1) feedstock processing: using poly glycol monomethyl ether (PEG1000) and lactide as raw material, by poly glycol monomethyl ether
It after being dissolved in methylene chloride, is removed water, is filtered spare with molecular sieve；Twice with ethyl alcohol recrystallization by lactide, drying for standby；
(2) synthesis of mPEG-b-PLA: pretreated 1g mPEG1000 is dissolved in 50mL methylene chloride, and 500r/min is stirred
It mixes to obtain mPEG1000 solution, 240 μ L DBN is added；9g lactide is dissolved in 50mL methylene chloride and obtains Lactide solution, then
Lactide solution is transferred in mPEG1000 solution, air is taken out, after reacting 180min, 110 μ l are added in vacuum degree 0.1Mpa
Glacial acetic acid terminates reaction, continues to stir 10min, obtains crude product after removing methylene chloride.Wherein, poly glycol monomethyl ether: third
The weight ratio of lactide is 1:9, and amidine class catalyst DBN additive amount is the 3% of reaction raw materials total mole number；The additive amount of terminator is
The 3% of reaction raw materials total mole number.
(3) column chromatographic purifying: after 2g crude product is dissolved with 10mL eluant, eluent (methylene chloride), use filler solidifying for glucan
The column chromatography for separation of glue LH60, and be rinsed by the above-mentioned eluant, eluent of 20 loading volumes.Main peak fraction is collected, after dry
White powder is obtained, nuclear-magnetism test is done.
Nuclear-magnetism result is solvent peak, the CH of mPEG at δ=1.8ppm as shown in Figure 1:2In the CH of δ=3.6ppm, PLA3With
For CH in δ=1.6ppm and δ=5.1ppm, integrated value ratio is 1.00:4.07:1.34, by characteristic peak integral than calculating to obtain copolymerization
Object structure is mPEG1000-b-PLA8848, the degree of polymerization 98.3%；Column chromatographic purifying: yield 93.5%, purity 99.5%.
Embodiment 2, a kind of preparation method of block copolymer mPEG-b-PLA (mPEG1000-b-PLA3000), including with
Lower step:
(1) feedstock processing: using poly glycol monomethyl ether (PEG1000) and lactide as raw material, by poly glycol monomethyl ether
It after being dissolved in methylene chloride, is removed water, is filtered spare with molecular sieve；Twice with ethyl alcohol recrystallization by lactide, drying for standby, wherein
Poly glycol monomethyl ether: the weight ratio of lactide is 1:3.
(2) synthesis of mPEG-b-PLA: pretreated 2g mPEG1000 is dissolved in 50mL methylene chloride, and 500r/min is stirred
It mixes to obtain mPEG1000 solution, 130 μ g TBD is added；6g lactide is dissolved in 50mL methylene chloride and obtains Lactide solution, then
Lactide solution is transferred in mPEG1000 solution, takes out air, and vacuum degree is that 0.09Mpa reacts 30min；89 μ g benzoic acid are added
Reaction is terminated, continues to stir 10min, obtains crude product after removing methylene chloride.Wherein, poly glycol monomethyl ether: lactide
Weight ratio is 1:3, and amidine class catalyst TBD additive amount is the 1.5% of reaction raw materials total mole number；The additive amount of terminator is reaction
The 1.5% of raw material total mole number.
(3) column chromatographic purifying: after 3g crude product is dissolved with 10mL eluant, eluent (ethyl alcohol: DCM=2:1), sephadex
The eluant, eluent of LH20 column chromatography for separation, 20 loading volumes is rinsed.Main peak fraction is collected, obtains transparent paste after dry
Object does nuclear-magnetism test.
Nuclear-magnetism result is as shown in Figure 2: the CH of copolymer structure, mPEG2In the CH of δ=3.6ppm, PLA3With CH δ=
1.6ppm and δ=5.1ppm, integrated value ratio be 1.00:1.21:0.38, by characteristic peak integral than calculate copolymer structure is
MPEG1000-b-PLA2642, the degree of polymerization 88.1%；Column chromatographic purifying: yield 94.4%, purity 99.5%；GPC result is such as
Shown in Fig. 3: being in Unimodal Distribution, dispersion degree 1.07 illustrates that the product only exists a kind of continuous polymer of molecular weight, and molecule
Amount distribution is very narrow.
Embodiment 3, a kind of preparation method of block copolymer mPEG-b-PLA (mPEG1000-b-PLA1000), including with
Lower step:
(1) feedstock processing: using poly glycol monomethyl ether (PEG1000) and lactide as raw material, by poly glycol monomethyl ether
It after being dissolved in methylene chloride, is removed water, is filtered spare with molecular sieve；Twice with ethyl alcohol recrystallization by lactide, drying for standby.
(2) synthesis of mPEG-b-PLA: pretreated 5g mPEG1000 is dissolved in 50mL methylene chloride and obtains mPEG1000
Solution, 500r/min stirring, is added 60 μ l DBU；5g lactide is dissolved in 50mL methylene chloride and obtains Lactide solution, then will
It obtains Lactide solution to be transferred in mPEG1000 solution, is passed through nitrogen after taking out air, reacts 40min under nitrogen protection；Add
Enter 36 μ l ethanedioic acids and terminate reaction, continues to stir 5min, obtain crude product after removing methylene chloride.Wherein, polyethyleneglycol first
Ether: the weight ratio of lactide is 1:1, and amidine class catalyst DBU additive amount is the 1% of reaction raw materials total mole number；Terminator adds
Dosage is the 1% of reaction raw materials total mole number.
(3) column chromatographic purifying: after 3g crude product is dissolved with 10mL eluant, eluent (methanol), use filler for sephadex
The column chromatography for separation of G50, and be rinsed by the eluant, eluent of 20 loading volumes.Main peak fraction is collected, is obtained after dry
Gelatin-like liquid does nuclear-magnetism test.
Nuclear-magnetism result is as shown in Figure 4: the CH of mPEG2In the CH of δ=3.6ppm, PLA3With CH δ=1.6ppm and δ=
5.1ppm, integrated value ratio be 1.00:0.42:0.13, by characteristic peak integral than calculate copolymer structure be mPEG1000-b-
PLA913, the degree of polymerization 91.30%；Column chromatographic purifying: yield 92.5%, purity 99.5%.
Embodiment 4, a kind of preparation method of block copolymer mPEG-b-PLA (mPEG5000-b-PLA3000), including with
Lower step:
(1) feedstock processing: using poly glycol monomethyl ether (PEG5000) and lactide as raw material, by poly glycol monomethyl ether
It after being dissolved in methylene chloride, is removed water, is filtered spare with molecular sieve；Twice with ethyl alcohol recrystallization by lactide, drying for standby.
(2) synthesis of mPEG-b-PLA:
Pretreated 5g mPEG5000 is dissolved in 50mL methylene chloride, and 500r/min stirs to get mPEG5000 solution, adds
Enter 70 μ g TBD；3g lactide is molten to obtain Lactide solution with 50mL methylene chloride, and then Lactide solution is transferred to mPEG5000
In solution, air is taken out, reacts 60min；35 μ l ethanedioic acids are added and terminate reaction, continue to stir 10min, after removing methylene chloride
Obtain crude product.Wherein, poly glycol monomethyl ether: the weight ratio of lactide is 5:3, and amidine class catalyst TBD additive amount is reaction
The 1.5% of raw material total mole number；The additive amount of terminator is the 1.5% of reaction raw materials total mole number.
(3) column chromatographic purifying: after 3g crude product is dissolved with 10mL eluant, eluent (ethyl alcohol), use filler for sephadex
The column chromatography for separation of G100, and be rinsed by the eluant, eluent ethyl alcohol of 20 loading volumes.Main peak fraction is collected, is obtained after dry
To white powder, nuclear-magnetism test is done.
Micella is made with o/w method in above-mentioned purification of samples, and 3000r/min is centrifuged 5min, takes supernatant to be lyophilized to get to finally
Purified product.
Nuclear-magnetism result is as shown in Figure 5: the CH of mPEG2In the CH of δ=3.6ppm, PLA3With CH δ=1.6ppm and δ=
5.1ppm, integral ratio are 1.00:0.28:0.09, and obtaining copolymer structure by characteristic peak ratio calculation is mPEG5000-b-
PLA2940, the degree of polymerization 98%；Column chromatographic purifying: yield 95.5%；GPC result is as shown in fig. 6, be in bimodal distribution, main peak point
Divergence 1.03；GPC result after centrifugation is as shown in fig. 7, be in Unimodal Distribution, dispersion degree 1.04, yield 85.5%, purity 99.5%.
It can be seen that: bimodal to become unimodal after centrifugal treating, by-product is separated, and dispersion degree is 1.04, illustrates to pass through
Cross centrifuge separation, which only remains a kind of continuous polymer of molecular weight, and disperse it is relatively narrow, it is therefore, simple for some
The product further separated can not be further realized using column chromatography, so that it may be centrifugated using micella is made.
Embodiment 5, a kind of preparation method of block copolymer mPEG-b-PLA (mPEG5000-b-PLA5000), including with
Lower step:
(1) feedstock processing: using poly glycol monomethyl ether (mPEG5000) and lactide as raw material, by poly glycol monomethyl ether
It after being dissolved in methylene chloride, is removed water, is filtered spare with molecular sieve；Twice with ethyl alcohol recrystallization by lactide, drying for standby.
(2) synthesis of mPEG-b-PLA: pretreated 5g mPEG5000 is dissolved in 50mL methylene chloride, and 500r/min is stirred
It mixes to obtain mPEG5000 solution, 140 μ g TBD is added；5g lactide is molten to obtain Lactide solution with 50mL methylene chloride, then
Lactide solution is transferred in mPEG5000 solution, takes out air, and vacuum degree 0.09Mpa reacts 70min；70 μ l second two are added
Acid terminates reaction, continues to stir 5min, obtains crude product after removing methylene chloride.Wherein, poly glycol monomethyl ether: lactide
Weight ratio is 1:1, and amidine class catalyst TBD additive amount is the 2% of reaction raw materials total mole number；The additive amount of terminator is that reaction is former
Expect the 2% of total mole number.
(3) column chromatographic purifying: after 3g crude product is dissolved with 10mL eluant, eluent (ethyl alcohol: DCM volume ratio=2:1), using filling out
Material is the column chromatography for separation of sephadex LH60, and the eluant, eluent of 20 loading volumes is rinsed.Main peak fraction is collected, is done
Nuclear-magnetism test is done after dry.
Micella is made with o/w method in above-mentioned purification of samples, and 2500r/min is centrifuged 5min, takes supernatant dry final pure to get arriving
Change product.
Nuclear-magnetism result is as shown in Figure 8: the CH of mPEG2In the CH of δ=3.6ppm, PLA3With CH δ=1.6ppm and δ=
5.1ppm, integral ratio are 1.00:0.41:0.13, are mPEG5000-b- by characteristic peak integral ratio calculation copolymer structure
PLA4476, the degree of polymerization 89.5%；Column chromatographic purifying: yield 95.8%；GPC result is in bimodal distribution, main peak dispersion degree
1.05；GPC result is in Unimodal Distribution, dispersion degree 1.05, yield 84.5%, purity 99.5% after centrifugation.
Embodiment 6, a kind of preparation method of block copolymer mPEG-b-PLA (mPEG5000-b-PLA8500), including with
Lower step:
(1) feedstock processing: using poly glycol monomethyl ether (PEG5000) and lactide as raw material, by poly glycol monomethyl ether
It after being dissolved in methylene chloride, is removed water, is filtered spare with molecular sieve；Twice with ethyl alcohol recrystallization by lactide, drying for standby.
(2) synthesis of mPEG-b-PLA: pretreated 5g mPEG3000 is dissolved in 50mL methylene chloride, and 500r/min is stirred
It mixes to obtain mPEG3000 solution, 220 μ g TBD is added；8.5g lactide is molten to obtain Lactide solution with 50mL methylene chloride, so
Lactide solution is transferred in mPEG3000 solution afterwards, takes out air, and vacuum degree 0.08Mpa reacts 40min；130 μ l fourths are added
Diacid terminates reaction, continues to stir 5min, obtains crude product after removing methylene chloride.Wherein, poly glycol monomethyl ether: lactide
Weight ratio be 1:1, amidine class catalyst loading be reaction raw materials total mole number 2%；The additive amount of terminator is that reaction is former
Expect the 2% of total mole number.
(3) column chromatographic purifying: after 3g crude product is dissolved with 10mL eluant, eluent (methylene chloride), sephadex LH60 column layer
Analysis separation, the eluant, eluent of 20 loading volumes are rinsed.Main peak fraction is collected, does nuclear-magnetism test after dry.
Micella is made with o/w method in above-mentioned purification of samples, and 2000r/min is centrifuged 5min, takes supernatant dry final pure to get arriving
Change product.
Nuclear-magnetism result is as shown in Figure 9: the CH of mPEG2In the CH of δ=3.6ppm, PLA3With CH δ=1.6ppm and δ=
5.1ppm, integral ratio are 1.00:0.74:0.24, and obtaining copolymer structure by characteristic peak ratio calculation is mPEG5000-b-
PLA8043, the degree of polymerization 94.6%；GPC result is in bimodal distribution, main peak dispersion degree 1.04；GPC result is in unimodal point after centrifugation
Cloth, divergence 1.05, yield 87.5%, purity 99.9%.
Embodiment 7, a kind of preparation method of block copolymer mPEG-b-PLA (mPEG1000-b-PLA10000), including
Following steps:
(1) feedstock processing: using poly glycol monomethyl ether (PEG1000) and lactide as raw material, by poly glycol monomethyl ether
It after being dissolved in acetone, is removed water, is filtered spare with molecular sieve；Twice with acetone recrystallization by lactide, drying for standby.
(2) synthesis of mPEG-b-PLA: pretreated 1g mPEG1000 is dissolved in 50mL acetone, and 500r/min is stirred
To mPEG1000 solution, 420 μ gTBD are added；10g lactide is dissolved in 50ml acetone and obtains Lactide solution, then will obtain third
Lactide solution is transferred in mPEG1000 solution, takes out air, is filled with nitrogen, reacts 40min；250 μ l benzoic acid and second two is added
Sour (volume ratio 10:1) terminates reaction, continues to stir 10min, obtains crude product after removing acetone.Wherein, polyethyleneglycol first
Ether: the weight ratio of lactide is 1:10, and amidine class catalyst loading is the 3% of reaction raw materials total mole number；The addition of terminator
Amount is the 3% of reaction raw materials total mole number.
(3) column chromatographic purifying: after 3g crude product is dissolved with 10mL eluant, eluent (chloroform), use filler solidifying for glucan
The column chromatography for separation of glue LH60, and be rinsed by the eluant, eluent of 20 loading volumes, main peak fraction is collected, is obtained after dry
White powder does nuclear-magnetism test.
Crude product nuclear-magnetism is as the result is shown: the CH of mPEG2In the CH of δ=3.6ppm, PLA3With CH δ=1.6ppm and δ=
5.1ppm, integral ratio are 1.00:4.03:1.43, and being integrated by characteristic peak than calculating the degree of polymerization is 94.0%；Column chromatographic purifying:
Yield 93.9%, purity 99.5%.
Embodiment 8, a kind of preparation method of block copolymer mPEG-b-PLA (mPEG6000-b-PLA200), including with
Lower step:
(1) feedstock processing: using poly glycol monomethyl ether (PEG6000) and lactide as raw material, by poly glycol monomethyl ether
It after being dissolved in chloroform, is removed water, is filtered spare with molecular sieve；Twice with ethyl alcohol recrystallization by lactide, drying for standby.
(2) synthesis of mPEG-b-PLA: pretreated 9g mPEG6000 is dissolved in 50mL chloroform, and 500r/min is stirred
It mixes to obtain mPEG6000 solution, 4.2 μ g TBD is added；0.3g lactide is molten to obtain Lactide solution with 50mL chloroform, so
After will obtain Lactide solution and be transferred in mPEG6000 solution, take out air, react 10min；2 μ l ethanedioic acids are added to terminate instead
It answers, continues to stir 5min, obtain crude product after removing chloroform.Wherein, poly glycol monomethyl ether: the weight ratio of lactide is
30:1, amidine class catalyst loading are the 0.1% of reaction raw materials total mole number；The additive amount of terminator is reaction raw materials total moles
Several 0.1%.
(3) column chromatographic purifying: after 3g crude product is dissolved with 10mL eluant, eluent (ethyl alcohol), 25 column of sephadex G chromatography point
From the eluant, eluent of 20 loading volumes is rinsed.Main peak fraction is collected, does nuclear-magnetism test after dry.
Nuclear-magnetism is as the result is shown: the CH of mPEG2In the CH of δ=3.6ppm, PLA3With CH δ=1.6ppm and δ=5.1ppm,
Basic ratio: 71.42:1.00:0.34 is mPEG6000-b-PLA190, the degree of polymerization by characteristic peak ratio calculation copolymer structure
It is 95%；Column chromatographic purifying: yield 90.5%, purity 99.5%.
Functional experiment of the block copolymer mPEG-b-PLA of the present invention as Thermosensitive Material Used for Controlled Releasing of Medicine:
Taxadol slow release experiment
1) standard curve
Taxol is configured to 40 μ g/ml, 20 μ g/ml, 10 μ g/ml, 5 μ g/ml, 2.5 μ g/ml, 1 μ g/ with dehydrated alcohol
The solution of ml tests ultraviolet absorptivity, obtains standard curve, regression equation: y=0.0232x
2) prepared by taxol micella
200mg mPEG5000-b-PLA3000,20mg taxol are dissolved in 2ml methylene chloride, 20ml is added to and goes
In ionized water, 140w ultrasound 15min is used under ice-water bath, rear room temperature 200r/min, which is stirred to organic solvent, to volatilize completely to get arriving
Taxol micella.
3) sustained release experiment
By above-mentioned micella, high speed refrigerated centrifuge 10min, washing are centrifuged again after precipitating under 4 DEG C, 15000rpm speed, weight
It washes twice again, merges all supernatants, measuring encapsulation rate is 90.1%, and centrifugation gained precipitating is dispersed with 10ml PBS buffer solution
In the bag filter of 3500D, bag filter is dialysed in 20mlPBS buffer, is placed in the constant temperature oscillator of 37 DEG C, 180r/min
In, respectively in 2h, 4h, 6h, 22h, 31h, 48h, 57h, 78h, 95 sampling 10ml dissolution fluids, 10mlPBS buffer is added, by sample
Product are extracted with dichloromethane respectively, then dry methylene chloride, are dissolved with 4ml ethyl alcohol, and test absorbance makes release profiles, release
Put curve such as Figure 10.Elution profiles illustrate that mPEG-b-PLA can be used for Thermosensitive Material Used for Controlled Releasing of Medicine.
Comparative experiments: weighing 5g mPEG5000 respectively, and 8.5g lactide is placed in together in the dry round-bottomed flask of 100ml,
15mg stannous octoate is heated to 140 DEG C of magnetic agitation reaction 8h, obtains faint yellow clear viscous fluid under vacuum conditions；
It is dissolved with methylene chloride, instills precipitating in anhydrous cold ether, stand overnight, leaching sediment is dried in vacuo at room temperature, is produced
Object.
The molecular weight structural of design is mPEG5000-b-PLA8500, nuclear-magnetism result such as Figure 11, and actual molecular weight structure is
MPEG5000-b-PLA6300, the degree of polymerization 74.1%, GPC result such as Figure 12 are in bimodal distribution, main peak dispersion degree 1.15.
It can be seen that preparation method of the invention is reacted at normal temperature after comparative experiments and embodiment 6 compare, the reaction time
The significantly shorter than prior art, and have the effect of accurately controlling the molecular weight and its molecular weight distribution of product, and have more
The high degree of polymerization, the degree of polymerization are up to 90% or more.
In addition, as can be seen from example 4 that, before and after centrifugal treating in GPC result, the bimodal distribution shown in Figure 12 becomes
The single distribution shown in Fig. 7, by-product are separated completely.Experiments have shown that: in purification process of the invention, chromatographic column can be by list
The separation of small molecuies such as body, catalyst, terminator；Centrifugal treating can separate the close with principal product molecular weight of side reaction generation
Polymer.Therefore, the present invention passes through two-step purifying, can be kept completely separate by-product, can high efficiency obtain Narrow Molecular Weight Distribution
The di-block copolymer mPEG-b-PLA of product.
Claims (9)
1. a kind of preparation method of block copolymer mPEG-b-PLA, which comprises the following steps:
(1) using poly glycol monomethyl ether and lactide as raw material, poly glycol monomethyl ether pretreatment of raw material: is dissolved in organic solvent
Afterwards, remove water, filter it is spare；By lactide recrystallization, drying for standby；
(2) synthesis of mPEG-b-PLA: poly glycol monomethyl ether and lactide are weighed, wherein poly glycol monomethyl ether: lactide
Weight ratio be (0.1~30): 1；Poly glycol monomethyl ether and lactide are dissolved with volatile organic solvent respectively and gathered
Ethylene glycol monomethyl ether solution and Lactide solution；Amidine class catalyst is added in poly glycol monomethyl ether solution, then under stiring
Lactide solution is added, polymerize 10-180min under oxygen free condition, reaction is then terminated using terminator, obtains crude product；Its
In, amidine class catalyst loading is the 0.1%~3% of reaction raw materials total mole number；The additive amount of terminator is that reaction raw materials are total
The 0.1%~3% of molal quantity；
(3) column chromatographic purifying: after crude product is dissolved with eluant, eluent, then by the isolated purified product of chromatographic column, wherein washing
De- agent is volatile organic solvent.
2. the preparation method of block copolymer mPEG-b-PLA as described in claim 1, which is characterized in that after step (3),
It is further comprising the steps of:
(4) o/w/ prepares micella: the purified product that column chromatography for separation is crossed is dissolved in volatile organic solvent, then mixes sufficiently with water
Emulsification, stirring are volatilized completely to volatile organic solvent, obtain micella；
(5) it is centrifuged: micella being subjected to centrifugal treating, takes supernatant that end product is lyophilized to obtain.
3. the preparation method of block copolymer mPEG-b-PLA as claimed in claim 1 or 2, which is characterized in that the step
(1), in (2), volatile organic solvent selects alkane, alkane substitute or the alcohol of low carbon chain, and the carbon chain lengths are C1-C10。
4. the preparation method of block copolymer mPEG-b-PLA as claimed in claim 1 or 2, which is characterized in that the step
(2) in, oxygen free condition is under vacuum or in inert gas.
5. the preparation method of block copolymer mPEG-b-PLA as claimed in claim 1 or 2, which is characterized in that the step
(2) in, the terminator is organic acid.
6. the preparation method of block copolymer mPEG-b-PLA as claimed in claim 1 or 2, which is characterized in that the amidine class
Catalyst is TBD, DBN or DBU.
7. the preparation method of block copolymer mPEG-b-PLA as claimed in claim 1 or 2, which is characterized in that the step
(3) in, the filler of the column chromatography is sephadex, and granularity is 100~400 mesh.
8. the preparation method of block copolymer mPEG-b-PLA as claimed in claim 7, which is characterized in that the step (3)
In, the filler of the column chromatography is the glucose gel of G type or LH type.
9. the preparation method of block copolymer mPEG-b-PLA as claimed in claim 1 or 2, which is characterized in that the step
(3) in, the eluant, eluent is methanol, ethyl alcohol, methylene chloride, chloroform or two or more of mixtures in them.
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| US20130324680A1 (en) * | 2010-11-30 | 2013-12-05 | Ningbo Institute Of Material Technology And Engineering, Chinese Academy Of Sciences | Polylactic acid block copolymers and preparation methods thereof |
| CN104761710A (en) * | 2014-02-14 | 2015-07-08 | 苏州海特比奥生物技术有限公司 | Methoxypolyethylene glycol-lactic acid block copolymer and preparation method thereof |
| CN106939075A (en) * | 2015-12-31 | 2017-07-11 | 银谷制药有限责任公司 | A kind of hydrophilic polyester and its block copolymer |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US20130324680A1 (en) * | 2010-11-30 | 2013-12-05 | Ningbo Institute Of Material Technology And Engineering, Chinese Academy Of Sciences | Polylactic acid block copolymers and preparation methods thereof |
| CN104761710A (en) * | 2014-02-14 | 2015-07-08 | 苏州海特比奥生物技术有限公司 | Methoxypolyethylene glycol-lactic acid block copolymer and preparation method thereof |
| US20160137775A1 (en) * | 2014-02-14 | 2016-05-19 | Suzhou High-Tech Bioscience Co., Ltd. | Methoxypolyethylene glycol-polylactic acid block copolymer and preparation method thereof |
| CN106939075A (en) * | 2015-12-31 | 2017-07-11 | 银谷制药有限责任公司 | A kind of hydrophilic polyester and its block copolymer |
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