CN110105550A - A kind of high-mechanical property biocompatible block copolymer and preparation method and application - Google Patents
A kind of high-mechanical property biocompatible block copolymer and preparation method and application Download PDFInfo
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- CN110105550A CN110105550A CN201910431983.7A CN201910431983A CN110105550A CN 110105550 A CN110105550 A CN 110105550A CN 201910431983 A CN201910431983 A CN 201910431983A CN 110105550 A CN110105550 A CN 110105550A
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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
- C08G63/02—Polyesters derived from hydroxycarboxylic acids or from polycarboxylic acids and polyhydroxy compounds
- C08G63/06—Polyesters derived from hydroxycarboxylic acids or from polycarboxylic acids and polyhydroxy compounds derived from hydroxycarboxylic acids
- C08G63/08—Lactones or lactides
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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
- C08G63/68—Polyesters containing atoms other than carbon, hydrogen and oxygen
- C08G63/685—Polyesters containing atoms other than carbon, hydrogen and oxygen containing nitrogen
- C08G63/6854—Polyesters containing atoms other than carbon, hydrogen and oxygen containing nitrogen derived from polycarboxylic acids and polyhydroxy compounds
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- C08G63/82—Preparation processes characterised by the catalyst used
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Abstract
The invention discloses a kind of high-mechanical property biocompatible block copolymer and preparation method and applications.It the described method comprises the following steps: (1) preparing hydroxy-end capped BPLP prepolymer；(2) L- lactide, glycolide, catalyst and BPLP prepolymer are reacted in a reservoir, the organic solvent being then evaporated in vacuo in catalyst seals container vacuum gas washing and flame, oil bath heating, cooling, purifying, vacuum filter, vacuum drying, obtains BPLP-co-PLGA copolymer.The copolymer combines the excellent properties of BPLP and PLGA, has excellent biocompatibility, meets bio-medical material requirement；And the thermomechanical property of adjustable final product is arranged by the amount of change BPLP and the ratio of components of LA and GA in copolymer synthesis process.
Description
Technical field
The invention belongs to the technical fields of biological medical polymer material, and in particular to a kind of high-mechanical property bio-compatible
Property copolymer and preparation method and application.
Background technique
Medical domain is studied and be applied to biological medical polymer material extensively, becomes medical product core
Core material；But this kind of material is difficult simultaneously both inanimate object cytotoxicities, and has visualization capability, while possessing again excellent
The ability of mechanical performance, this kind of material is very rare in a word.Currently, existing method be by biodegradable polymer with
Semiconductor-quantum-point or organic dyestuff mixing are conjugated to generate the Biodegradable material of luminescence generated by light.But semiconductor amount
Son point is at high cost, and the intrinsic toxicity with heavy metal；And organic dyestuff has photobleaching and lower dyestuff-poly-
Substance markers ratio is closed, so that it is easily separated from the drug of load, these problems all hinder the practical application of the method.There are also methods just
It is to use green fluorescent protein (GFP) as the substitute for showing inherent luminescence generated by light, but due to protein overexpression and gather
Collection, it would still be possible to cause cytotoxicity, it is also difficult to reach requirement.
Currently, there are also a kind of biodegradable light emitting polymer (BPLP) bio-medical materials with inherent luminescence generated by light
Material is able to satisfy numerous requirements, it has adjustable fluorescence, high quantum production rate and excellent photostability.But BPLP polymer by
In its low molecular weight (1200Da), it is provided with the property of low viscosity, so as to cause its easily aggregation in physiological conditions, and works as it
When heat cross-linking is biodegradable elastomer, its tensile strength is for many organizational projects using not high enough.On its basis
The copolymer (BPLP-PLLA) that the BPLP (UBPLP) and BPLP and L- lactide of upper improved carbamate doping are copolymerized is all
There is certain improvement, but the rigidity and brittleness of PLLA polymer are bad, and degradation is slowly, and BPLP-PLLA polymer
Synthesis in use enzyme-catalyzed polymerization method, these cannot all cause its large-scale practical application.
The above several method can only be generated with one or several satisfactory polymer, cannot be applied completely
In clinical test.But it is still considerable that research, which has the biodegradable light emitting polymer (BPLP) of inherent luminescence generated by light,
BPLP is included in polylactide-glycolic acid poly and closed in object (PLGA) by direction, generate it is a kind of new there is inherent luminescence generated by light can
Biodegradable light-emitting copolymers (BPLP-co-PLGA), since PLGA has good biocompatibility, faster biology drop
Xie Xing, outstanding mechanical performance and the features such as being readily synthesized, in many excellent properties of fusion BPLP, so that copolymer
BPLP-co-PLGA is able to satisfy the extensive large-scale application requirement of biological medical polymer material.
Summary of the invention
In order to overcome the bio-incompatibility of biological medical polymer material, cytotoxicity and bad machine in the prior art
The disadvantages of tool performance and deficiency, the object of the present invention is to provide a kind of high-mechanical property biocompatible block copolymer and preparation methods
With application.
The purpose of the present invention is achieved through the following technical solutions.
The present invention provides a kind of preparation methods of high-mechanical property biocompatible block copolymer, comprising the following steps:
(1) in protective gas, polybasic carboxylic acid, polyalcohol and amino acid are added in container 1, stirs, is heated to
T1, until polybasic carboxylic acid, polyalcohol and the amino acid fusing in container 1, it is cooled to T2, in T2At a temperature of continue to stir it is several
Time obtains prepolymer；
(2) prepolymer obtained with Isosorbide-5-Nitrae-dioxanes dissolving step (1) obtains pre-polymer solution, then is carried out with deionized water
Prepolymer after purification is freeze-dried, obtains hydroxy-end capped pure BPLP prepolymer by purifying；
(3) L- lactide, glycolide and BPLP prepolymer are added in anhydrous methylene chloride, are then added in container 2, with
After catalyst is added, be heated to 160-170 DEG C carry out reaction 8-12 hours；
(4) anhydrous methylene chloride is evaporated in vacuo, by 2 vacuum gas washing of container, flame sealing, container 2, which is put into temperature, is
T3Oil bath in some time, be subsequently cooled to environment temperature, obtain solid matter, pure solid substance is that is, molten by solid matter
Solution obtains mixed solution 1 in chloroform, then mixed solution 1 is instilled in dehydrated alcohol, obtains sediment, repeats the operation 3-5 of purifying
Secondary, vacuum filter, vacuum drying obtains BPLP-co-PLGA copolymer.
Preferably, the polyalcohol in step (1) is α, ω-normal alkane glycol；The amino acid is cysteine, silk ammonia
One or more of acid, glycine；The polybasic carboxylic acid is one or more of citric acid, citrate；
The polybasic carboxylic acid, polyalcohol and amino acid molar ratio are (1~2): (1.1~2): (0.2~1)；
The container 1 is three neck round bottom；
The protective gas is nitrogen；
T1It is 155~165 DEG C；T2It is 135~145 DEG C, in T2At a temperature of continue stirring 1-1.5 hours.
Preferably, Isosorbide-5-Nitrae-dioxanes mass fraction is 20%-30% in step (2) pre-polymer solution；
The quality of deionized water used is the 30%-40% of the quality of the pre-polymer solution；
The time of freeze-drying is 24~25 hours.
Preferably, the molar ratio of L- lactide, glycolide and BPLP prepolymer is (1-3): 1:(0.01- in step (3)
0.08)；
Catalyst is the stannous 2-ethylhexoate Sn (Oct) that purity is 94-96%2, stannous 2-ethylhexoate Sn (Oct)2's
Quality is the 0.1-0.15% of both L- lactide and glycolide quality sum；L- lactide, glycolide and BPLP prepolymer three
Moles total number and anhydrous methylene chloride mass ratio be (6.7-10.2) mmol/g.
Preferably, in step (4), the time of vacuum evaporation is 0.5~1 hour；Container 2 is drying reaction tube；Vacuum is washed
Gas, which refers to, to be vacuumized container 2 and is purged 3-5 times with nitrogen；T3It is 155~165 DEG C；It is T that container 2, which is put into temperature,3Oil bath
In time be 48~49 hours；The mass fraction of chloroform is 30%-40% in mixed solution 1；The quality of dehydrated alcohol is mixed
Close the 40%-50% of the quality of solution 1；The vacuum drying time is 7~10 days.
The present invention also provides the BPLP-co- of the preparation method of high-mechanical property biocompatible block copolymer preparation
PLGA copolymer.
The present invention also provides the BPLP-co-PLGA copolymers to prepare answering in BPLP-co-PLGA co-polymer membrane
Include: that BPLP-co-PLGA copolymer is dissolved in absolute dichloromethane with the step of, application, obtains mixed solution 2, then will mixing
Solution 2 is cast in polytetrafluoroethylene PTFE container, evaporates methylene chloride；BPLP-co-PLGA copolymer in mixed solution 2
Concentration is 8-13mg/ml.
The present invention also provides the BPLP-co-PLGA copolymers to prepare the application in BPLP-co-PLGA bracket, answers
With the step of include: that BPLP-co-PLGA copolymer is dissolved in Isosorbide-5-Nitrae-dioxanes, mixed solution 3 is obtained, in mixed solution 3
The screening salt having a size of 50-250 μm is added, evaporates Isosorbide-5-Nitrae-dioxanes, leaches screening salt with deionized water, sample is obtained, by sample
Product freeze-drying；The time of the freeze-drying is 24~25 hours；The concentration of BPLP-co-PLGA copolymer in mixed solution 3
For 8-13mg/ml；The mass ratio for sieving salt and BPLP-co-PLGA copolymer is 8-10.
The present invention also provides the BPLP-co-PLGA copolymers in preparing BPLP-co-PLGA nano particle
Include: that 1. BPLP-co-PLGA copolymer is dissolved in chloroform using the step of, application, obtains mixed solution 4；2. will mix molten
Liquid 4 is added drop-wise in PVA aqueous solution and is ultrasonically treated simultaneously, obtains oil-in-water emulsion；3. oil-in-water emulsion is stirred, chloroform is evaporated；
4. being centrifuged, wash, freeze-drying obtains BPLP-co-PLGA nano particle；
The concentration of BPLP-co-PLGA copolymer is 20~30mg/ml in mixed solution 4；
The mass volume ratio of the PVA aqueous solution is 4-6%mg/ml；
The volume ratio of mixed solution 4 and PVA aqueous solution is 1:(8~10)；
3. the middle time stirred is 24~25 hours to step；4. middle washing is to be washed with distilled water 3-5 times to step.
The present invention also provides the BPLP-co-PLGA copolymers in the BPLP-co- for preparing 5 FU 5 fluorouracil load
The step of application in PLGA nano particle, application includes: that BPLP-co-PLGA copolymer is dissolved in chloroform by (a), is obtained mixed
Close solution 5；(b) 5 FU 5 fluorouracil aqueous solution is added in mixed solution 5, and be ultrasonically treated simultaneously, obtain Water-In-Oil W/O lotion；
(c) ultrasound homogenizing Water-In-Oil W/O lotion, the Water-In-Oil W/O lotion after homogenizing is added in PVA aqueous solution, W/O/W is obtained
W/O/W lotion；(d) W/O/W W/O/W lotion is stirred, evaporates chloroform；(e) it is centrifuged, washs, freeze-drying obtains 5- fluorine
The BPLP-co-PLGA nano particle of uracil load；
The mass volume ratio of the PVA aqueous solution is 4-6%mg/ml；
The concentration of BPLP-co-PLGA copolymer is 20~30mg/ml in mixed solution 5；
The concentration of 5 FU 5 fluorouracil aqueous solution is 5-5.5mg/mL；
The volume ratio of 5 FU 5 fluorouracil aqueous solution and mixed solution 5 is 1:(2~3)；Mixed solution 5 and PVA aqueous solution
Volume ratio is 1:(8~10)；The time of step (d) stirring is 24~25 hours；Step (e) washing is to be washed with distilled water 3-5
It is secondary.
Compared with prior art, the present invention has the following advantages and beneficial effects:
The present invention is to be passed through by the BPLP prepolymer that first synthesis of hydroxy blocks then as macromole evocating agent
Stannous 2-ethylhexoate (Sn (Oct)2) catalysis ring-opening polymerization, react, obtain with L- lactide (LA) and glycolide (GA)
To BPLP-co-PLGA copolymer.PLGA, which is added, makes it combine the excellent properties of BPLP and PLGA, has excellent biofacies
Capacitive meets bio-medical material requirement；And pass through the amount and LA and GA for changing BPLP in copolymer synthesis process
Ratio of components arranges the thermomechanical property of adjustable final product.
Detailed description of the invention
Fig. 1 is the pure PLGA polymerization of BPLP-co-PLGA copolymer prepared by embodiment 2-7 and embodiment 2-3 preparation
The infrared spectrogram (ATR-FTIR) of object；
Fig. 2 is to prepare pure PLGA in the pure BPLP-co-PLGA copolymer and embodiment 2-3 prepared in embodiment 2-7 to gather
Close the tensile strength and Young's modulus comparison diagram of object；
Fig. 3 is to polymerize the pure BPLP-co-PLGA copolymer in embodiment 2,4,5,6 and the pure PLGA in embodiment 2
Object preparation film forming, after stem cell is then cultivated 24 hours above it, the fluorescence property comparison diagram of corresponding membrane.
Specific embodiment
Below with reference to embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited to
This.
Embodiment 1
Present embodiments provide the preparation method of pure prepolymer BPLP, comprising the following steps:
(1) under constant nitrogen stream, the citric acid, 1,8- ethohexadiol and the cysteine that are 1:1.1:0.2 by molar ratio
It is added in three neck round bottom, stirs, be heated to 160 DEG C, until citric acid, 1,8- ethohexadiol and cysteine fusing, cooling
To 140 DEG C, 140 DEG C at a temperature of continue stirring 1 hour, obtain prepolymer；
(2) prepolymer obtained with Isosorbide-5-Nitrae-dioxanes dissolving step (1), obtains pre-polymer solution, Isosorbide-5-Nitrae-in pre-polymer solution
The mass fraction of dioxanes is 25%, and pre-polymer solution is deposited in deionized water dropwise, to remove unreacted citric acid,
1,8- ethohexadiol and cysteine purify gained prepolymer, and the quality of deionized water used is above-mentioned pre-polymer solution
35%, prepolymer after purification is lyophilized 24 hours in freeze-dryer, obtains pure prepolymer BPLP.
Embodiment 2
(1) a kind of preparation method of high-mechanical property biocompatible block copolymer, including following step are present embodiments provided
It is rapid:
The BPLP prepolymer of the L- lactide of 0.06mol, the glycolide of 0.02mol and 0.0016mol is added to 10g's
In anhydrous methylene chloride, in being added to drying reaction tube, stannous 2-ethylhexoate Sn (Oct) then is added2(Sn(Oct)2's
Purity is 95%, Sn (Oct)2Quality be the 0.12% of both L- lactide and glycolide quality summation), be warming up to 165 DEG C,
Reaction 10 hours, then methylene chloride is evaporated in vacuo 1 hour, drying reaction tube is vacuumized and is purged three times with nitrogen, flame
Sealing will be dried reaction tube and be put into the oil bath that temperature is 160 DEG C 48 hours, is subsequently cooled to environment temperature, obtains solid matter,
Solid matter is purified again, i.e., solid matter is dissolved in chloroform, mixed solution 1 is obtained, the quality of chloroform point in mixed solution 1
Number is 35%, then mixed solution 1 is instilled in dehydrated alcohol and is precipitated, and the quality of dehydrated alcohol is the 45% of 1 mass of mixed solution,
It repeats purifying three times, then vacuum filter, is dried in vacuo 7 days at room temperature, obtains BPLP-co-PLGA copolymer, i.e. BPLP-co-
PLGA-1。
(2) the present embodiment also contrastively proposes the preparation method of pure PLGA polymer, preparation step and raw material dosage
(1) preparation step of the BPLP-co-PLGA-1 in is identical with raw material dosage, the difference is that BPLP is not added in round-bottomed flask
Prepolymer is prepared into pure PLGA polymer, i.e. PLGA-1.
It is manufactured in the present embodiment that there is inherent luminescence generated by light and biodegradable medical macromolecular materials BPLP-co-
PLGA-1 and PLGA-1 for carrying out tension test, measure its tensile strength and Young's modulus and by BPLP-co-PLGA-1 and
PLGA-1 preparation film forming, obtains BPLP-co-PLGA-1 film and PLGA-1 film and then cultivates stem cell above it 24 hours
Afterwards, fluorescence detection is carried out again to corresponding film.
The infrared spectrogram of BPLP-co-PLGA-1 and PLGA-1 is shown in Fig. 1, it can be seen that in 1745cm-1There is suction at place
Receive peak, it is known that have ester group generation, it was demonstrated that BPLP-co-PLGA-1 is with the structure of PLGA-1 similar, and BPLP is only accounted for
The sub-fraction of BPLP-co-PLGA-1.
The tensile strength and Young's modulus figure of BPLP-co-PLGA-1 and PLGA-1 is shown in Fig. 2, it can be seen that PLGA-1 film
Tensile strength and Young's modulus will be much higher than BPLP-co-PLGA-1 film, illustrate the mechanical performance ratio BPLP- in PLGA-1 film
Co-PLGA-1 film will be got well, and with the increase of PLGA content, the mechanical performance of film is also increasing, and illustrate that PLGA improves BPLP-
The mechanical performance of co-PLGA-1.
BPLP-co-PLGA-1 and pure PLGA-1 preparation film forming is right after stem cell is then cultivated 24 hours above it
The fluorescence detection of film is as shown in figure 3, measuring the comparable cell survival rate on all films by mtt assay and passing through SEM in BPLP-
The stretching form of cell on co-PLGA-1 film shows that BPLP-coPLGA supports human mesenchymal stem cell (hMSC) adherency and increases
Cytotoxicity is grown and shows, the fluorescence intensity ratio PLGA-1 of BPLP-co-PLGA-1 wants high as seen from the figure, illustrates its
Cytotoxicity is more preferable, cell can be allowed preferably to survive, to know that BPLP-co-PLGA-1 biological property is fine.
Embodiment 3
(1) a kind of preparation method of high-mechanical property biocompatible block copolymer, including following step are present embodiments provided
It is rapid:
The BPLP prepolymer of the L- lactide of 0.04mol, the glycolide of 0.04mol and 0.0016mol is added to 10g's
In anhydrous methylene chloride, in being added to drying reaction tube, stannous 2-ethylhexoate Sn (Oct) then is added2(Sn(Oct)2's
Purity is 95%, Sn (Oct)2Quality be the 0.12% of both L- lactide and glycolide quality summation), be warming up to 165 DEG C,
Reaction 10 hours, then methylene chloride is evaporated in vacuo 1 hour, drying reaction tube is vacuumized and is purged three times with nitrogen, flame
Sealing will be dried reaction tube and be put into the oil bath that temperature is 160 DEG C 48 hours, is subsequently cooled to environment temperature, obtains solid matter,
Solid matter is purified again, i.e., solid matter is dissolved in chloroform, mixed solution 1 is obtained, the quality of chloroform point in mixed solution 1
Number is 35%, then mixed solution 1 is instilled in dehydrated alcohol and is precipitated, and the quality of dehydrated alcohol is the 45% of 1 mass of mixed solution,
It repeats purifying three times, then vacuum filter, is dried in vacuo 7 days at room temperature, obtains BPLP-co-PLGA copolymer, i.e. BPLP-co-
PLGA-2。
(2) the present embodiment also contrastively proposes the preparation method of pure PLGA polymer, preparation step and raw material dosage
(1) preparation step of the BPLP-co-PLGA-2 in is identical with raw material dosage, the difference is that BPLP is not added in round-bottomed flask
Prepolymer is prepared into pure PLGA polymer, i.e. PLGA-2.
It is manufactured in the present embodiment that there is inherent luminescence generated by light and biodegradable medical macromolecular materials BPLP-co-
PLGA-2 is for examination of infrared spectrum and carries out tension test, measures its tensile strength and Young's modulus.
The infrared spectrogram of BPLP-co-PLGA-2 and PLGA-2 can refer to Fig. 1 in embodiment 2；
The tensile strength and Young's modulus figure of BPLP-co-PLGA-2 and PLGA-2 is shown in Fig. 2, it can be seen that PLGA-1 film
Tensile strength and Young's modulus to be much higher than PLGA-2 film, the tensile strength and Young's modulus of PLGA-1 film be higher than BPLP-
The film of co-PLGA-2, and the tensile strength and Young's modulus of PLGA-2 film are higher than the film of BPLP-co-PLGA-2, illustrate LA and
The ratio of GA influences the mechanical performance of copolymer, the i.e. mechanical performance of the adjustable copolymer of amount of change LA and GA.
Embodiment 4
Present embodiments provide a kind of preparation method of high-mechanical property biocompatible block copolymer, comprising the following steps:
The BPLP prepolymer of the L- lactide of 0.06mol, the glycolide of 0.02mol and 0.0008mol is added to 10g's
In anhydrous methylene chloride, in being added to drying reaction tube, stannous 2-ethylhexoate Sn (Oct) then is added2(Sn(Oct)2's
Purity is 95%, Sn (Oct)2Quality be the 0.12% of both L- lactide and glycolide quality summation), be warming up to 165 DEG C,
Reaction 10 hours, then methylene chloride is evaporated in vacuo 1 hour, drying reaction tube is vacuumized and is purged three times with nitrogen, flame
Sealing will be dried reaction tube and be put into the oil bath that temperature is 160 DEG C 48 hours, is subsequently cooled to environment temperature, obtains solid matter,
Solid matter is purified again, i.e., solid matter is dissolved in chloroform, mixed solution 1 is obtained, the quality of chloroform point in mixed solution 1
Number is 35%, then mixed solution 1 is instilled in dehydrated alcohol and is precipitated, and the quality of dehydrated alcohol is the 45% of 1 mass of mixed solution,
It repeats purifying three times, then vacuum filter, is dried in vacuo 7 days at room temperature, obtains BPLP-co-PLGA copolymer, i.e. BPLP-co-
PLGA-3。
It is manufactured in the present embodiment that there is inherent luminescence generated by light and biodegradable medical macromolecular materials BPLP-co-
PLGA-3 is for examination of infrared spectrum and carries out tension test, measures its tensile strength and Young's modulus and by pure BPLP-co-
PLGA-3 preparation film forming, obtains BPLP-co-PLGA-3 film, after stem cell is then cultivated 24 hours above it, to the fluorescence of film
Detection.
The infrared spectrogram of BPLP-co-PLGA-3 can refer to Fig. 1 in embodiment 2；
BPLP-co-PLGA-3 tensile strength and Young's modulus figure such as Fig. 2, it can be seen that the tensile strength of PLGA-1 film
It is slightly above BPLP-co-PLGA-3 film with Young's modulus, but the tensile strength and Young's modulus of BPLP-co-PLGA-3 film want high
In PLGA-2 and BPLP-co-PLGA-1 film, illustrate that the ratio of LA and GA influences the mechanical performance of copolymer.
The biological property of BPLP-co-PLGA-3 can refer to the performance of BPLP-co-PLGA-1 prepared in embodiment 2.
Embodiment 5
Present embodiments provide a kind of preparation method of high-mechanical property biocompatible block copolymer, comprising the following steps:
(1) the BPLP prepolymer of the L- lactide of 0.04mol, the glycolide of 0.04mol and 0.0008mol is added to
In the anhydrous methylene chloride of 10g, in being added to drying reaction tube, stannous 2-ethylhexoate Sn (Oct) then is added2(Sn
(Oct)2Purity be 95%, Sn (Oct)2Quality be the 0.12% of both L- lactide and glycolide quality summation), heating
It to 165 DEG C, reacts 10 hours, then methylene chloride is evaporated in vacuo 1 hour, drying reaction tube is vacuumized and purge three with nitrogen
Secondary, flame sealing will be dried reaction tube and be put into the oil bath that temperature is 160 DEG C 48 hours, is subsequently cooled to environment temperature, must consolidate
Body substance, then solid matter is purified, i.e., solid matter is dissolved in chloroform, obtains mixed solution 1, chloroform in mixed solution 1
Mass fraction be 35%, then will mixed solution 1 instill dehydrated alcohol in precipitate, the quality of dehydrated alcohol is 1 matter of mixed solution
The 45% of amount repeats purifying three times, then vacuum filter, is dried in vacuo 7 days at room temperature, obtains BPLP-co-PLGA copolymer,
That is BPLP-co-PLGA-4.
It is manufactured in the present embodiment that there is inherent luminescence generated by light and biodegradable medical macromolecular materials BPLP-co-
PLGA-4 is for examination of infrared spectrum and carries out tension test, measures its tensile strength and Young's modulus and by BPLP-co-
PLGA-4 preparation film forming, obtains BPLP-co-PLGA-4 film, after stem cell is then cultivated 24 hours above it, to BPLP-co-
The fluorescence detection of PLGA-4 film.
The infrared spectrogram of BPLP-co-PLGA-4 can refer to Fig. 1 in embodiment 2；
The tensile strength and Young's modulus figure of BPLP-co-PLGA-4 is shown in Fig. 2, it can be seen that the stretching of PLGA-1 film is strong
Degree and Young's modulus are much higher than BPLP-co-PLGA-4 film, but the tensile strength and Young's modulus of BPLP-co-PLGA-4 film are wanted
Higher than PLGA-2 and BPLP-co-PLGA-2 film, illustrate that the ratio of LA and GA influences the mechanical performance of copolymer.
The biological property of BPLP-co-PLGA-4 can refer to the performance of BPLP-co-PLGA-1 prepared in embodiment 2.
Embodiment 6
Present embodiments provide a kind of preparation method of high-mechanical property biocompatible block copolymer, comprising the following steps:
(1) the BPLP prepolymer of the L- lactide of 0.06mol, the glycolide of 0.02mol and 0.0004mol is added to
In the anhydrous methylene chloride of 10g, in being added to drying reaction tube, stannous 2-ethylhexoate Sn (Oct) then is added2(Sn
(Oct)2Purity be 95%, Sn (Oct)2Quality be the 0.12% of both L- lactide and glycolide quality summation), heating
It to 165 DEG C, reacts 10 hours, then methylene chloride is evaporated in vacuo 1 hour, drying reaction tube is vacuumized and purge three with nitrogen
Secondary, flame sealing will be dried reaction tube and be put into the oil bath that temperature is 160 DEG C 48 hours, is subsequently cooled to environment temperature, must consolidate
Body substance, then solid matter is purified, i.e., solid matter is dissolved in chloroform, obtains mixed solution 1, chloroform in mixed solution 1
Mass fraction be 35%, then will mixed solution 1 instill dehydrated alcohol in precipitate, the quality of dehydrated alcohol is 1 matter of mixed solution
The 45% of amount repeats purifying three times, then vacuum filter, is dried in vacuo 7 days at room temperature, obtains BPLP-co-PLGA copolymer,
That is BPLP-co-PLGA-5.
It is manufactured in the present embodiment that there is inherent luminescence generated by light and biodegradable medical macromolecular materials BPLP-co-
PLGA-5 is for examination of infrared spectrum and carries out tension test, measures its tensile strength and Young's modulus and by BPLP-co-
PLGA-5 preparation film forming, obtains BPLP-co-PLGA-5 film, after stem cell is then cultivated 24 hours above it, to the fluorescence of film
Detection.
The infrared spectrogram of BPLP-co-PLGA-5 can refer to Fig. 1 in embodiment 2；
The tensile strength and Young's modulus figure of BPLP-co-PLGA-5 is shown in Fig. 2, it can be seen that the stretching of PLGA-1 film is strong
Degree and Young's modulus are comparable to BPLP-co-PLGA-5 film, but the tensile strength and Young's modulus of BPLP-co-PLGA-5 film are wanted
Higher than BPLP-co-PLGA-1 and BPLP-co-PLGA-3 film, illustrate that the ratio of LA and GA influences the mechanical performance of copolymer.
The biological property of BPLP-co-PLGA-5 can refer to the performance of BPLP-co-PLGA-1 prepared in embodiment 2.
Embodiment 7
Present embodiments provide a kind of preparation method of high-mechanical property biocompatible block copolymer, comprising the following steps:
The BPLP prepolymer of the L- lactide of 0.04mol, the glycolide of 0.04mol and 0.0004mol is added to 10g's
In anhydrous methylene chloride, in being added to drying reaction tube, stannous 2-ethylhexoate Sn (Oct) then is added2(Sn(Oct)2's
Purity is 95%, Sn (Oct)2Quality be the 0.12% of both L- lactide and glycolide quality summation), be warming up to 165 DEG C,
Reaction 10 hours, then methylene chloride is evaporated in vacuo 1 hour, drying reaction tube is vacuumized and is purged three times with nitrogen, flame
Sealing will be dried reaction tube and be put into the oil bath that temperature is 160 DEG C 48 hours, is subsequently cooled to environment temperature, obtains solid matter,
Solid matter is purified again, i.e., solid matter is dissolved in chloroform, mixed solution 1 is obtained, the quality of chloroform point in mixed solution 1
Number is 35%, then mixed solution 1 is instilled in dehydrated alcohol and is precipitated, and the quality of dehydrated alcohol is the 45% of 1 mass of mixed solution,
It repeats purifying three times, then vacuum filter, is dried in vacuo 7 days at room temperature, obtains BPLP-co-PLGA copolymer, obtain BPLP-co-
PLGA-6。
It is manufactured in the present embodiment that there is inherent luminescence generated by light and biodegradable medical macromolecular materials BPLP-co-
PLGA-6 is for examination of infrared spectrum and carries out tension test, measures its tensile strength and Young's modulus.
The infrared spectrogram of BPLP-co-PLGA-6 can refer to Fig. 1 in embodiment 2；
The tensile strength and Young's modulus figure of BPLP-co-PLGA-6 is shown in Fig. 2, it can be seen that the stretching of PLGA-1 film is strong
Degree and Young's modulus are much higher than BPLP-co-PLGA-6 film, but the tensile strength and Young's modulus of BPLP-co-PLGA-6 film are wanted
Higher than BPLP-co-PLGA-2 and BPLP-co-PLGA-4 film, illustrate that the ratio of LA and GA influences the mechanical performance of copolymer.
Embodiment 8
A kind of BPLP-co-PLGA copolymer is present embodiments provided in preparation BPLP-co-PLGA co-polymer membrane, BPLP-
Answering in the BPLP-co-PLGA nano particle of co-PLGA bracket, BPLP-co-PLGA nano particle and 5 FU 5 fluorouracil load
With applying step includes:
(1) BPLP-co-PLGA-1 of 100mg is dissolved in 10ml absolute dichloromethane, obtains mixed solution 2, then will mixing
Solution 2 is cast in polytetrafluoroethyldisk disk, is evaporated methylene chloride, is obtained BPLP-co-PLGA copolymer-1 film.
(2) BPLP-co-PLGA-1 of 100mg is dissolved in Isosorbide-5-Nitrae-dioxanes of 10ml, obtains mixed solution 3, then will mix
Close the screening salt for being added that the size of 900mg is 150 μm in solution 3.Isosorbide-5-Nitrae-dioxanes is evaporated, leaches screening salt with deionized water,
Freeze-drying 24 hours, obtains BPLP-co-PLGA bracket.
(3) 100mg BPLP-co-PLGA-1 is dissolved in 5ml chloroform, obtains mixed solution 4, then by mixed solution 4
It is added drop-wise in the PLA aqueous solution of 40mL (5%mg/ml), and is ultrasonically treated simultaneously to generate oil-in-water emulsion, oil-in-water is being changed
It learns in draught cupboard and stirs 24 hours to evaporate chloroform, the particle as obtained by being collected by centrifugation, and be washed with distilled water three times, it is then cold
It is lyophilized dry, obtains BPLP-co-PLGA nano particle.
(4) 100mg BPLP-co-PLGA-1 is dissolved in 5mL chloroform, obtains mixed solution 5, by 2mL 5 FU 5 fluorouracil
Aqueous solution (5mg/mL) is slowly added to be ultrasonically treated in mixed solution 5 and simultaneously, obtains primary water-in-oil emulsion；Ultrasound homogenizing is primary
Water-in-oil emulsion is simultaneously slowly added in the PVA aqueous solution of 40mL (5%mg/ml), and water-in-oil-in-water compositions are obtained；By oil-in-water
Packet aqueous emulsion stirs 24 hours in chemical hood to evaporate chloroform；The particle as obtained by being collected by centrifugation, and washed with distillation
It washs three times, freeze-drying obtains the BPLP-co-PLGA nano particle of 5 FU 5 fluorouracil load.
Above several case study on implementation are the preferable several embodiments of the present invention, described in the above content to illustrate the invention
Basic principle, therefore the present invention is not limited to the above embodiments.Under the premise of basic principle and range of the invention, can also
There is various change and improve, these changes and improvements both fall within scope of the claimed invention.
Claims (10)
1. a kind of preparation method of high-mechanical property biocompatible block copolymer, which comprises the following steps:
(1) in protective gas, polybasic carboxylic acid, polyalcohol and amino acid are added in container 1, stirs, is heated to T1, directly
To polybasic carboxylic acid, polyalcohol and the amino acid fusing in container 1, it is cooled to T2, in T2At a temperature of continue stir some time,
Obtain prepolymer；
(2) prepolymer obtained with Isosorbide-5-Nitrae-dioxanes dissolving step (1), obtains pre-polymer solution, then purified with deionized water,
By prepolymer freeze-drying after purification, hydroxy-end capped pure BPLP prepolymer is obtained；
(3) L- lactide, glycolide and BPLP prepolymer are added in anhydrous methylene chloride, are then added in container 2, then added
Enter catalyst, is heated to 160-170 DEG C and carries out reaction 8-12 hours；
(4) anhydrous methylene chloride is evaporated in vacuo, by 2 vacuum gas washing of container, flame sealing, it is T that container 2, which is put into temperature,3Oil
Some time in bath, it is subsequently cooled to environment temperature, obtains solid matter, solid matter is dissolved in chlorine by pure solid substance
In imitative, mixed solution 1 obtained, then mixed solution 1 is instilled in dehydrated alcohol, obtain sediment, repeat operation 3-5 times of purifying, very
Sky filtering, vacuum drying obtain BPLP-co-PLGA copolymer.
2. the preparation method of high-mechanical property biocompatible block copolymer according to claim 1, which is characterized in that step
(1) polyalcohol in is α, ω-normal alkane glycol；The amino acid be one of cysteine, serine, glycine with
On；The polybasic carboxylic acid is one or more of citric acid, citrate；The polybasic carboxylic acid, polyalcohol and amino acid mole
Than for (1 ~ 2): (1.1 ~ 2): (0.2 ~ 1)；The container 1 is three neck round bottom；The protective gas is nitrogen；T1For
155~165℃；T2It is 135 ~ 145 DEG C, in T2At a temperature of continue stirring 1-1.5 hours.
3. the preparation method of high-mechanical property biocompatible block copolymer according to claim 1, which is characterized in that step
(2) mass fraction of 1,4- dioxanes is 20%-30% in pre-polymer solution；The quality of deionized water used is that the prepolymer is molten
The 30%-40% of the quality of liquid；The time of freeze-drying is 24 ~ 25 hours.
4. the preparation method of high-mechanical property biocompatible block copolymer according to claim 1, which is characterized in that step
(3) molar ratio of L- lactide, glycolide and BPLP prepolymer is (1-3): 1:(0.01-0.08 in)；Catalyst is that purity is
The stannous 2-ethylhexoate Sn (Oct) of 94-96%2, stannous 2-ethylhexoate Sn (Oct)2Quality be L- lactide and glycolide
The 0.1-0.15% of the two quality sum；The moles total number of L- lactide, glycolide and BPLP prepolymer three and anhydrous dichloromethane
The mass ratio of alkane is (6.7-10.2) mmol/g.
5. the preparation method of high-mechanical property biocompatible block copolymer according to claim 1, which is characterized in that step
(4) in, the time of vacuum evaporation is 0.5 ~ 1 hour；Container 2 is drying reaction tube；Vacuum gas washing, which refers to, vacuumizes container 2 simultaneously
It is purged 3-5 times with nitrogen； T3It is 155 ~ 165 DEG C；It is T that container 2, which is put into temperature,3Oil bath in time be 48 ~ 49 hours；It is mixed
The mass fraction for closing chloroform in solution 1 is 30%-40%；The quality of dehydrated alcohol is the 40%-50% of the quality of mixed solution 1；Very
The sky dry time is 7 ~ 10 days.
6. the BPLP- of the preparation method preparation of any one of claim 1 to the 5 high-mechanical property biocompatible block copolymer
Co-PLGA copolymer.
7. BPLP-co-PLGA copolymer as claimed in claim 6 is preparing the application in BPLP-co-PLGA co-polymer membrane,
It being characterized in that, the step of application includes: that BPLP-co-PLGA copolymer is dissolved in absolute dichloromethane, mixed solution 2 is obtained, then
Mixed solution 2 is cast in polytetrafluoroethylene PTFE container, methylene chloride is evaporated；BPLP-co-PLGA is total in mixed solution 2
The concentration of polymers is 8-13mg/ml.
8. BPLP-co-PLGA copolymer as claimed in claim 6 is preparing the application in BPLP-co-PLGA bracket, feature
It is, the step of application includes: that BPLP-co-PLGA copolymer is dissolved in Isosorbide-5-Nitrae-dioxanes, obtains mixed solution 3, is mixing
The screening salt having a size of 50-250 μm is added in solution 3, evaporates Isosorbide-5-Nitrae-dioxanes, leaches screening salt with deionized water, obtains sample
Sample is freeze-dried by product；The time of the freeze-drying is 24 ~ 25 hours；BPLP-co-PLGA copolymer in mixed solution 3
Concentration be 8-13mg/ml；The mass ratio for sieving salt and BPLP-co-PLGA copolymer is 8-10.
9. BPLP-co-PLGA copolymer as claimed in claim 6 is preparing the application in BPLP-co-PLGA nano particle,
It is characterized in that, the step of application includes: that 1. BPLP-co-PLGA copolymer is dissolved in chloroform, obtains mixed solution 4；2. will mix
It closes solution 4 to be added drop-wise in PVA aqueous solution and be ultrasonically treated simultaneously, obtains oil-in-water emulsion；3. oil-in-water emulsion is stirred, chlorine is evaporated
It is imitative；4. being centrifuged, wash, freeze-drying obtains BPLP-co-PLGA nano particle；BPLP-co-PLGA copolymer in mixed solution 4
Concentration be 20 ~ 30 mg/ml；The mass volume ratio of the PVA aqueous solution is 4-6%mg/ml；Mixed solution 4 and PVA are water-soluble
The volume ratio of liquid is 1:(8 ~ 10)；3. the middle time stirred is 24 ~ 25 hours to step；4. middle washing is to be washed with distilled water to step
3-5 times.
10. BPLP-co-PLGA copolymer as claimed in claim 6 is received in the BPLP-co-PLGA for preparing 5 FU 5 fluorouracil load
Application in rice grain, which is characterized in that the step of application includes: that BPLP-co-PLGA copolymer is dissolved in chloroform by (a),
Obtain mixed solution 5；(b) 5 FU 5 fluorouracil aqueous solution is added in mixed solution 5, and be ultrasonically treated simultaneously, obtain Water-In-Oil W/O cream
Liquid；(c) ultrasound homogenizing Water-In-Oil W/O lotion, the Water-In-Oil W/O lotion after homogenizing is added in PVA aqueous solution, oil-in-water packet is obtained
Water W/O/W lotion；(d) W/O/W W/O/W lotion is stirred, evaporates chloroform；(e) it is centrifuged, washs, freeze-drying obtains 5-
The BPLP-co-PLGA nano particle of fluorouracil load；
The mass volume ratio of the PVA aqueous solution is 4-6% mg/ml；
The concentration of BPLP-co-PLGA copolymer is 20 ~ 30 mg/ml in mixed solution 5；
The concentration of 5 FU 5 fluorouracil aqueous solution is 5-5.5mg/mL；The volume ratio of 5 FU 5 fluorouracil aqueous solution and mixed solution 5 is
1:(2 ~ 3)；The volume ratio of mixed solution 5 and PVA aqueous solution is 1:(8 ~ 10)；The time of step (d) stirring is 24 ~ 25 hours；
Step (e) washing is to be washed with distilled water 3-5 times.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1555278A1 (en) * | 2004-01-15 | 2005-07-20 | Innocore Technologies B.V. | Biodegradable multi-block co-polymers |
| CN105555833A (en) * | 2013-07-09 | 2016-05-04 | 德克萨斯大学体系董事会 | Fluorescent polymers and applications thereof |
-
2019
- 2019-05-23 CN CN201910431983.7A patent/CN110105550A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1555278A1 (en) * | 2004-01-15 | 2005-07-20 | Innocore Technologies B.V. | Biodegradable multi-block co-polymers |
| CN105555833A (en) * | 2013-07-09 | 2016-05-04 | 德克萨斯大学体系董事会 | Fluorescent polymers and applications thereof |
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| Title |
|---|
| JIANQING HU ET AL.: "Fluorescence imaging enabled poly(lactide-co-glycolide)", 《ACTA BIOMATERIALIA》 * |
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