CN107744503A - The preparation method of the amphipathic polyester MePEG Peptide PER CL administrations nanoparticle of enzyme sensitiveness - Google Patents
The preparation method of the amphipathic polyester MePEG Peptide PER CL administrations nanoparticle of enzyme sensitiveness Download PDFInfo
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Abstract
The invention belongs to pharmaceutical carrier macromolecule polymer material technical field, a kind of preparation method of amphipathic polyester of enzyme sensitiveness (MePEG Peptide PER CL) nanoparticle is disclosed.Its preparation is carried out in two steps：1) preparation of amphipathic polyester (MePEG Peptide PER CL)；2) curcumin mixes in proportion with MePEG Peptide PER CL, adds acetone solution and forms lipid phase；The surfactants such as polysorbate85, PLURONICS F87 (P 188), polyvinyl alcohol (PVA) and dodecyl sodium sulfate are dissolved in water and form aqueous phase；Lipid phase is slowly instilled into aqueous phase, 0.5~8h is stirred by emulsion-solvent evaporation method, prepares nanoparticle.Purpose obtains 100~280nm of average grain diameter, and polydispersity index (PDI) is less than 0.30, and envelop rate is more than 65% curcumin nanoparticles.Such nanometer of drug administration carrier can be used as a kind of load curcumin slow-released carrier system, have the advantages that targeting is strong, bioavilability is high, toxic side effect is small, suitable for a variety of administering modes such as intravenous injection, oral.
Description
(1) technical field
The present invention relates to the preparation of medicine macromolecule carrier and technical field of medicine, and in particular to a kind of enzyme sensitiveness two
The preparation method of nanoparticle is administered in parent's property polyester MePEG-Peptide-PER-CL.
(2) background technology
Cancer is one of global maximum public health problem, governs the development of social economy, greatly endangers people
The life of class, and by as the first killer of new century human health.Therefore, antineoplastic objects system efficiently, less toxic is found
Had very important significance for capturing tumor disease.
Curcumin (Curcumin) be extracted from the rhizome of some plants in Zingiber, Araeceae one kind chemistry into
Point, wherein, turmeric is the very rare pigment with diketone of plant kingdom containing about 3%~6%, is cyclohexadione compounds.Medical science is ground
Study carefully and show, curcumin has the effect such as reducing blood lipid, antitumor, anti-inflammatory, cholagogic, anti-oxidant.But curcumin solubility is low, external
Oxidizable, easily metabolism in vivo, causes its bioavilability low, limits its use in clinic, these characteristics limit it
Clinical practice.By researching and developing the novel form of curcumin, the clinical practice of curcumin can be promoted.
Nanoparticle (Nanoparticles) refers to particle diameter in 10~1000nm, using solid natural or the lipoid of synthesis to carry
Body, medicine is wrapped up or clamps the manufactured solid micelle delivery system in lipoid core, with toxicity is low, good biocompatibility, life
The solid natural of Biodegradable or the lipoid of synthesis are carrier, by Drug absorbability or are wrapped in manufactured a new generation in lipid film and receive
Grain of rice delivery system, have the advantages that can with Drug controlled release, avoid degraded or leakage and the good targeting of medicine.
Studies have reported that：Using nano molecular as gene transfer and the carrier of medicine, medicine, DNA and protein equimolecular are wrapped in
Among nano molecular, safely and effectively medicine and gene therapy can be finally realized.
Matrix metalloproteinase (matrix metalloproteinases, MMP) is that extracellular matrix degradation process can not
A kind of conservative enzyme lacked, is widely present in animal and plant body.But (such as inflammation, tumour when lesion occurs for human body
Fine raw infiltration, transfer and recurrence), the expression of matrix metalloproteinase (MMP) will significantly rise.Therefore, with curcumin
For medicine, MMP specific cleavages peptide is modified amphipathic polymer support, and curcumin can be achieved and released in tumor locus targeting
Put, so as to play the purpose for improving drug effect.
Amphipathilic block polymer refers to both there is very long hydrophobic chain segment in molecule, there is very long hydrophilic segment again
High molecular polymer, also known as high molecular surfactant.In field of medicaments, amphipathic multi-arm star-type polymer has good
Good drug encapsulation ability, biocompatibility and biodegradability, and it is new such as receptor target to be obtained by base group modification
Function, medicine can be avoided to be removed in vivo by internal organs and immunocyte, reduce drug toxicity to the lethality of normal cell, carry
The high security of medication.
(3) content of the invention
It is an object of the invention to provide a kind of amphipathic polyester MePEG-Peptide-PER-CL administrations nanometer of enzyme sensitiveness
The preparation method of grain.
The present invention adopts the following technical scheme that：
A kind of preparation method of the amphipathic polyester MePEG-Peptide-PER-CL administrations nanoparticle of enzyme sensitiveness, it is described
Preparation method is：
(1) amphipathic polyester MePEG-Peptide-PER-CL preparation：
1. PER-CL preparation：N2Under protection, by 136.2mg (1.0mmol) pentaerythrite, 6.8464g (60.0mmol)
6-caprolactone, the mixing of 60 μ g (0.15mol) stannous octoates, 140 DEG C of reaction 72h are warming up to, room temperature are cooled to, by reactant mixture
After being dissolved with 15mL dichloromethane, it is added drop-wise in 250mL ether, Precipitation, precipitation is collected by filtration, is purified through recrystallizing methanol
Secondary, at room temperature, 24h is to constant weight for vacuum drying, saves backup；
2. MePEG-COOH preparation：Take 1.9000g (1mmol) poly glycol monomethyl ether MePEG1900 and 0.1200g
(1.2mmol) succinic anhydride is added in 50ml three-necked flask, adds the stirring of 20ml pyridines until whole dissolvings.Keep nitrogen stream
It is logical, 0.1000g 4- dimethylamino naphthyridines DMAP, 20 μ L triethylamines are rapidly joined, 24h is reacted at room temperature after sealing.Product is turned
Move on in 25ml constant pressure funnel, be added drop-wise to 250ml ice ether dropwise, make precipitation abundant, filter.Product is placed in 40
DEG C vacuum drying chamber dries 24h to constant weight, saves backup；
3. MePEG-NHS preparation：Weigh above-mentioned 1. MePEG-COOH 1.9820g (1.0 mmol) and N- hydroxysuccinimidyls
Acid imide is placed in 50ml round-bottomed flask by 0.1381g (1.2mmol), adds the quick stirring and dissolving of 15mL acetonitriles.Add
0.1060g DCC (N, N- dicyclohexylcarbodiimide), react at room temperature 24h.Reaction solution is added drop-wise in 150ml ice ether,
It is added dropwise, makes precipitation abundant, filter, the filter cake being filtrated to get is product, and product is placed in into 40 DEG C of vacuum drying chambers dries
24h saves backup to constant weight.
4. MePEG-Peptide preparation：Under inert gas shielding, by Peptide ((ACP)-
GPLGIAGQrrrrrrrrr- (ACP)), n-hydroxysuccinimide (DMAP), 1- ethyls-(3- dimethyl chloros propyl group) carbon
Diimmonium salt hydrochlorate (EDC) mixes, with acetonitrile: water (v: v) 20: 80 is dissolved completely, and 2h is activated under ice bath；Dissolved with acetonitrile
Above-mentioned 2. MePEG-NHS, under stirring at room temperature, MePEG-NHS solution is added dropwise under inert gas shielding, reacts 2~5d, obtain and slightly produce
Thing solution A.
Product A purifying method be：By crude product in solution A, molecule M is placed inWIn 3500 bag filter, to use dialysis clamp
Sealing two ends.It is placed in 500ml distilled water.Preceding 5 hours per first water is changed every other hour, changed one every two hours afterwards
Secondary distilled water.Rotating speed 100r/min, room temperature dialysis 3d.Dialysis collects liquid in bag filter after terminating, vacuum freeze drying 24h
To constant weight, purified product MePEG-Peptide is produced；
Step 4. in, the Peptide ((ACP)-GPLGIAGQrrrrrrrrr- (ACP)) passes through the limited public affairs of biology of shining by force
Department's purchase obtains (content 98.69%)；
The Peptide ((ACP)-GPLGIAGQrrrrrrrrr- (ACP)), n-hydroxysuccinimide (DMAP), 1-
Ethyl-(3- dimethyl chloros propyl group) carbodiimide hydrochloride (EDC), MePEG-NHS inventory mol ratio for 1: 10~
40: 10~40: 0.5~2.0；
5. MePEG-Peptide-PER-CL preparation：Under inert gas shielding, by above-mentioned 3. MePEG-Peptide, N-
HOSu NHS (DMAP), N, N- dicyclohexylcarbodiimides (DCC) mixing, are dissolved completely, in ice bath with dichloromethane
Lower activation 2h；With dichloromethane dissolving pentaerythrite-caprolactone (PER-CL), under stirring at room temperature, dripped under inert gas shielding
Add PER-CL solution, react 2~5d, obtain crude product in solution B；
Product B purifying method be：By crude product in solution B, molecule M is placed inWIn 14000 bag filter, to use dialysis clamp
Sealing two ends.It is placed in 500ml distilled water.Preceding 5 hours per first water is changed every other hour, changed one every two hours afterwards
Secondary distilled water.Rotating speed 100r/min, room temperature dialysis 3d.Dialysis collects liquid in bag filter after terminating, vacuum freeze drying 24h
To constant weight, the amphipathic polyester MePEG-Peptide-PER-CL of purified product is produced.
The MePEG-Peptide, n-hydroxysuccinimide (DMAP), N, N- dicyclohexylcarbodiimides (DCC), season
The mol ratio of the object amount that feeds intake of penta tetrol-caprolactone (PER-CL) is 1: 5~20: 5~20: 0.25~1；
In the present invention, amphipathic polyester MePEG-Peptide-PER-CL is calculated by gpc analysis made from step (1)
Mean molecule quantity and polydispersity coefficient (PDI), and calculated yield of weighing.
GPC conditions：Mobile phase：Tetrahydrofuran (1ml/min)；Detection temperature：35℃；Polymer is used for GPC detectable concentrations
For 30mg/ml；Sample size：50μL；Column type number：HP Phenogel guard column attached to a Phenogel
linear(2)5μ GPC column。
The amphipathic polyester MePEG-Peptide-PER-CL molecular weight ranges of gained are 17150~23104, its polydispersion
Coefficient (PDI) is less than 1.3, generally 1.05~1.28.
(2) MePEG-Peptide-PER-CL made from curcumin, step (1) is dissolved in organic solvent, forms fat
Phase；Surfactant is soluble in water, form aqueous phase；Gained lipid phase is added drop-wise in aqueous phase, stirs 0.5~8h, collects filtrate
The solution of the as described amphipathic polyester MePEG-Peptide-PER-CL administrations nanoparticle of enzyme sensitiveness；
The curcumin concentration is 0.6~1.0mg/mL, preferably 0.8mg/mL；
The molecular weight of the amphipathic polyester MePEG-Peptide-PER-CL is 17150~23104, preferably
MW22494；
The surfactant is selected from Tween-85, PVA, P-188, preferably dodecyl sodium sulfate, dodecyl sodium sulfonate
Sodium；
The mixing time is 0.5~8h, preferably 4h；
In step (2), the curcumin can be commercially available by conventional route.
The solution of the final amphipathic polyester MePEG-Peptide-PER-CL administrations nanoparticle of gained is determined by particle instrument
Nanoparticle particle diameter and more breadth coefficients (PDI), HPLC calculate turmeric cellulose content in solution, and computational envelope rate (EE).
W1：Total curcumin quality in nanoparticle solution；
W2：Nanoparticle solution is after 15000r/min is centrifuged, contained curcumin quality in supernatant.
Amphipathic polyester MePEG-Peptide-PER-CL administration nanoparticles average grain diameter 100 produced by the present invention~
280nm, polydispersity index (PDI) are less than 0.30, and envelop rate is more than 65%.
Relative to prior art, the beneficial effects of the present invention are：The amphipathic polyester MePEG- of the enzyme responsive type
Peptide-PER-CL delivery systems with digestion peptide its with selectively targeted, the toxic side effect of medicine can be reduced；It is hydrophilic
Fragment MePEG has the function that the passive target for escaping internal reticuloendothelial system or macrophage system is removed, and increase carrier is in blood
Residence time in liquid, improve pharmacokinetics；Lipophilic moiety PER-CL improves dissolubility of the curcumin in water, improves medicine
Bioavilability；The delivery system has preferable slow release effect, suitable for a variety of administering modes such as intravenous injection, oral.
(4) illustrate
Fig. 1：MePEG-Peptide-PER-CL synthetic routes.
(5) embodiment
Below by specific embodiment, technical scheme is further elaborated.
MePEG-Peptide is synthesized
Embodiment 1
Under inert gas shielding, 100.0mg (0.04mmol) Peptide ((ACP)-GPLGIAGQrrrrrrrrr- are weighed
(ACP)), 13.9mg (0.16mmol) n-hydroxysuccinimide (DMAP), 21.8mg (0.16mmol) 1- ethyls-(3- diformazans
Base chloro propyl group) carbodiimide hydrochloride (EDC) mixing, use acetonitrile：The dissolving of water (v: v) 20: 80 is complete, is activated under ice bath
2h；89.6mg (0.04mmol) MePEG-NHS is dissolved with acetonitrile, under stirring at room temperature, MePEG- is added dropwise under inert gas shielding
NHS solution, 2d is reacted, obtains crude product in solution A.Through MW3500 dialysis 3d, remove unreacted raw material, collect bag filter after purification
In solution, vacuum freeze drying 24h to constant weight, 144.7mg MePEG-Peptide are made.
It is 4833, PI 1.12 through GPC mean molecule quantities, ultimate yield 66.30%.
Embodiment 2
Under inert gas shielding, 100.0mg (0.04mmol) Peptide ((ACP)-GPLGIAGQrrrrrrrrr- are weighed
(ACP)), 48.9mg (0.40mmol) n-hydroxysuccinimide (DMAP), 76.7mg (0.40mmol) 1- ethyls-(3- diformazans
Base chloro propyl group) carbodiimide hydrochloride (EDC) mixing, use acetonitrile：The dissolving of water (v: v) 20: 80 is complete, is activated under ice bath
2h；125.9mg (0.06mmol) MePEG-NHS is dissolved with acetonitrile, under stirring at room temperature, MePEG- is added dropwise under inert gas shielding
NHS solution, 3d is reacted, obtains crude product in solution A.Through MW3500 dialysis 3d, remove unreacted raw material, collect bag filter after purification
In solution, vacuum freeze drying 24h to constant weight, 141.4mg MePEG-Peptide are made.
It is 5822, PI 1.02 through GPC mean molecule quantities, ultimate yield 74.59%.
Embodiment 3
Under inert gas shielding, 100.0mg (0.04mmol) Peptide ((ACP)-GPLGIAGQrrrrrrrrr- are weighed
(ACP)), 97.8mg (0.80mmol) n-hydroxysuccinimide (DMAP), 153.4mg (0.80mmol) 1- ethyls-(3- diformazans
Base chloro propyl group) carbodiimide hydrochloride (EDC) mixing, use acetonitrile：Water (v: v) 20：80 dissolvings are complete, activated under ice bath
2h；179.2mg (0.08mmol) MePEG-NHS is dissolved with acetonitrile, under stirring at room temperature, MePEG- is added dropwise under inert gas shielding
NHS solution, 5d is reacted, obtains crude product in solution A.Through MW3500 dialysis 3d, remove unreacted raw material, collect bag filter after purification
In solution, vacuum freeze drying 24h to constant weight, 153.8mg MePEG-Peptide are made.
It is 6588, PI 1.16 through GPC mean molecule quantities, ultimate yield 81.14%.
MePEG-Peptide-PER-CL preparation
Embodiment 4
By 232.9mg (0.04mmol) MePEG-Peptide, 24.4mg (0.20 mmol) N- hydroxysuccinimidyl acyls of example 2
Imines (DMAP), 41.3mg (0.20mmol) N, N- dicyclohexylcarbodiimide (DCC) mixing, are dissolved completely with dichloromethane,
2h is activated under ice bath；140.0mg (0.01) pentaerythrite-caprolactone (PER-CL) is dissolved with dichloromethane, is stirred at room temperature
Under, PER-CL solution is added dropwise under inert gas shielding, reacts 2d, obtains crude product in solution B；Through MW7000 dialysis 3d, are removed not anti-
The raw material answered, the solution in bag filter is collected after purification, vacuum freeze drying 24h to constant weight, 232.4mg MePEG- are made
Peptide-PER-CL。
It is 17150, PI 1.36 through GPC mean molecule quantities, ultimate yield 62.32%.
Embodiment 5
By 232.9mg (0.04mmol) MePEG-Peptide, 48.8mg (0.40 mmol) N- hydroxysuccinimidyl acyls of example 2
Imines (DMAP), 82.6mg (0.40mmol) N, N- dicyclohexylcarbodiimide (DCC) mixing, are dissolved completely with dichloromethane,
2h is activated under ice bath；280.0mg (0.02) pentaerythrite-caprolactone (PER-CL) is dissolved with dichloromethane, is stirred at room temperature
Under, PER-CL solution is added dropwise under inert gas shielding, reacts 3d, obtains crude product in solution B；Through MW7000 dialysis 3d, are removed not anti-
The raw material answered, the solution in bag filter is collected after purification, vacuum freeze drying 24h to constant weight, 299.9mg MePEG- are made
Peptide-PER-CL。
It is 22494, PI 1.13 through GPC mean molecule quantities, ultimate yield 80.42%.
Embodiment 6
By 232.9mg (0.04mmol) MePEG-Peptide, 97.6mg (0.80 mmol) N- hydroxysuccinimidyl acyls of example 2
Imines (DMAP), 165.2mg (0.80mmol) N, N- dicyclohexylcarbodiimide (DCC) mixing, are dissolved completely with dichloromethane,
2h is activated under ice bath；560.0mg (0.04) pentaerythrite-caprolactone (PER-CL) is dissolved with dichloromethane, is stirred at room temperature
Under, PER-CL solution is added dropwise under inert gas shielding, reacts 3d, obtains crude product in solution B；Through MW7000 dialysis 3d, are removed not anti-
The raw material answered, the solution in bag filter is collected after purification, vacuum freeze drying 24h to constant weight, 284.5mg MePEG- are made
Peptide-PER-CL。
It is 21626, PI 1.27 through GPC mean molecule quantities, ultimate yield 76.29%.
Embodiment 7
By 232.9mg (0.04mmol) MePEG-Peptide, 97.6mg (0.80 mmol) N- hydroxysuccinimidyl acyls of example 2
Imines (DMAP), 165.2mg (0.80mmol) N, N- dicyclohexylcarbodiimide (DCC) mixing, are dissolved completely with dichloromethane,
2h is activated under ice bath；560.0mg (0.04) pentaerythrite-caprolactone (PER-CL) is dissolved with dichloromethane, is stirred at room temperature
Under, PER-CL solution is added dropwise under inert gas shielding, reacts 5d, obtains crude product in solution B；Through MW7000 dialysis 3d, are removed not anti-
The raw material answered, the solution in bag filter is collected after purification, vacuum freeze drying 24h to constant weight, 290.6mg MePEG- are made
Peptide-PER-CL。
It is 23104, PI 1.18 through GPC mean molecule quantities, ultimate yield 77.93%.
It is prepared by the amphipathic polyester MePEG-Peptide-PER-CL curcumin nanoparticles of enzyme sensitiveness
Embodiment 8
Examine or check curcumin concentration influences on nanoparticle granulating.
Curcumin 6.0,8.0,10.0mg are settled to 10mL volumetric flasks with acetone solvent, are configured to the turmeric of various concentrations
Plain organic solution, is saved backup；2mL is dissolved according to MePEG-Peptide-PER-CL 24.0mg made from the method for embodiment 5
In curcumin acetone solvent, lipid phase is formed；15.0mg surfactant sodium dodecyl base sodium sulfonates are dissolved in 10mL water, form water
Phase；Gained lipid phase is added drop-wise in aqueous phase, stirs 4h, removes organic solvent, 6000r/min centrifugation 20min, removing is not encapsulated
Curcumin, collect supernatant be MePEG-Peptide-PER-CL curcumin nanoparticles solution；
As a result Table1 is seen
Table1 curcumins concentration influences on nanoparticle granulating
It is preferred that curcumin concentration 0.8mg/ml.
Embodiment 9
Examine or check curcumin/MePEG-Peptide-PER-CL ratios influences on nanoparticle granulating.
Curcumin 6.0mg is settled to 10mL volumetric flasks with acetone solvent, the curcumin for being configured to 0.6mg/mL is organic molten
Liquid, save backup；Weighed respectively according to the starlike polyester of MePEG-Peptide-PER-CL made from embodiment 4-7 methods
36.0mg is dissolved in 2mL curcumin acetone solvents, forms lipid phase；15.0mg surfactant sodium dodecyl base sodium sulfonates are dissolved in
In 10mL water, aqueous phase is formed；Gained lipid phase is added drop-wise in aqueous phase, stirs 4h, removes organic solvent, 6000r/min centrifugations
20min, non-encapsulated curcumin is removed, it is MePEG-Peptide-PER-CL curcumin nanoparticles solution to collect supernatant；
As a result Table2 is seen
Table2 curcumin MePEG-Peptide-PER-CL ratios influence on nanoparticle granulating
It is preferred that MePEG-Peptide-PER-CL molecular weight 22494.
Embodiment 10
Curcumin 6.0mg is settled to 10mL volumetric flasks with acetone solvent, the curcumin for being configured to 0.6 mg/mL is organic molten
Liquid, save backup；36.0mg is weighed respectively according to the starlike polyester of MePEG-Peptide-PER-CL made from the method for embodiment 5
It is dissolved in 2mL curcumin acetone solvents, forms lipid phase；By 15.0mg surface active agent tweens -85, P-188, PVA, dodecyl
Sodium sulfonate is dissolved in 10mL water, forms aqueous phase；Gained lipid phase is added drop-wise in aqueous phase, stirs 4h, removes organic solvent, 6000r/
Min centrifuges 20min, removes non-encapsulated curcumin, and it is MePEG-Peptide-PER-CL curcumin nanoparticles to collect supernatant
Solution；
As a result Table3 is seen
The selection of Table3 surfactants influences on nanoparticle granulating
It is preferred that dodecyl sodium sulfate.
Embodiment 11
Curcumin 8.0mg is settled to 10mL volumetric flasks with acetone solvent, the curcumin for being configured to 0.6 mg/mL is organic molten
Liquid, save backup；36.0mg is weighed respectively according to the starlike polyester of MePEG-Peptide-PER-CL made from the method for embodiment 5
It is dissolved in 2mL curcumin acetone solvents, forms lipid phase；15.0mg surfactant sodium dodecyl base sodium sulfonates are dissolved in 10mL water
In, form aqueous phase；Gained lipid phase is added drop-wise in aqueous phase, stirs 0.5,1,2,4,6,8 h, removes organic solvent, 6000r/min
20min is centrifuged, removes non-encapsulated curcumin, it is that MePEG-Peptide-PER-CL curcumin nanoparticles are molten to collect supernatant
Liquid；
As a result Table4 is seen
The Table4 reaction time influences on nanoparticle granulating
Preferred reaction time is 4h.
Claims (7)
1. a kind of preparation method of the amphipathic polyester MePEG-Peptide-PER-CL administrations nanoparticle of enzyme sensitiveness, its feature exist
In described preparation method is：
1) amphipathic polyester MePEG-Peptide-PER-CL preparation：
1. pentaerythrite and 6-caprolactone add appropriate stannous octoate, N in 140 DEG C of meltings2Under protective condition, ring-opening polymerisation 72h, lead to
Cross secondarily purified process and PER-CL monomers are made；
2. poly glycol monomethyl ether MePEG1900 and the appropriate pyridinium dissolution of succinic anhydride, add a certain amount of 4- dimethylaminos
Pyridine DMAP and triethylamine, N2 protections are lower to be reacted at room temperature, and clarification, obtains product MePEG-COOH；
3. MePEG-COOH and n-hydroxysuccinimide are dissolved with appropriate acetonitrile, in N, the catalysis of N- dicyclohexylcarbodiimides
Under, N2Under protection, 24h is reacted at room temperature, clarification, obtains product MePEG-NHS；
4. Peptide, n-hydroxysuccinimide and 1- ethyls-(3- dimethyl chloros propyl group) carbodiimide hydrochloride mixing are used
Appropriate acetonitrile and water mixed solution dissolving, 2h is activated under ice bath；Add MePEG-NHS, under stirring at room temperature, N2Under protection,
2~5d is reacted, clarification, obtains product MePEG-Peptide；
5. by above-mentioned MePEG-Peptide, n-hydroxysuccinimide DMAP, N, the mixing of N- dicyclohexylcarbodiimides is with right amount
Dichloromethane dissolving, activate 2h under ice bath；Pentaerythrite-caprolactone of dichloromethane dissolving is added, under stirring at room temperature,
N2Protection 2~5d of lower reaction, clarification, obtains product MePEG-Peptide-PER-CL；
Described 4. middle Peptide, n-hydroxysuccinimide, 1- ethyls-(3- dimethyl chloros propyl group) carbodiimide hydrochloride
Mol ratio with MePEG-NHS inventory is 1: (10~40): (10~40): (0.5~2.0)；
Described 5. middle MePEG-Peptide, n-hydroxysuccinimide, N, N- dicyclohexylcarbodiimides and pentaerythrite-oneself
The mol ratio of the object amount that feeds intake of lactone is 1: (5~20): (5~20): (0.25~1)；
2) the amphipathic polyester of enzyme sensitiveness made from curcumin, step (1) is dissolved in acetone, forms lipid phase；By surface-active
Agent forms aqueous phase in water；Gained lipid phase is added drop-wise in aqueous phase, stirs 0.5~8h, vacuum filtration, it is institute to collect filtrate
State the solution of the amphipathic polyester administration nanoparticle of enzyme sensitiveness；
The curcumin concentration is 0.6~1.0mg/mL；
The molecular weight of the amphipathic polyester MePEG-Peptide-PER-CL is 17150~23104；
The surfactant is selected from Tween-85, PVA, P-188 or dodecyl sodium sulfate.
2. the preparation of the amphipathic polyester MePEG-Peptide-PER-CL administrations nanoparticle of enzyme sensitiveness as claimed in claim 1
Method, it is characterised in that step 4. in, the Peptide, n-hydroxysuccinimide, 1- ethyls-(3- dimethyl chloro third
Base) the carbodiimide hydrochloride and MePEG-NHS mol ratio of inventory is 1: 10: 10: 1.5.
3. the preparation of the amphipathic polyester MePEG-Peptide-PER-CL administrations nanoparticle of enzyme sensitiveness as claimed in claim 1
Method, it is characterised in that step 5. in, the MePEG-Peptide, n-hydroxysuccinimide, N, N- dicyclohexyls carbon two
The mol ratio of the object amount that feeds intake of imines and pentaerythrite-caprolactone is 1: 10: 10: 0.5.
4. the preparation of the amphipathic polyester MePEG-Peptide-PER-CL administrations nanoparticle of enzyme sensitiveness as claimed in claim 1
Method, it is characterised in that in step 2), the curcumin concentration is 0.8mg/mL.
5. the preparation of the amphipathic polyester MePEG-Peptide-PER-CL administrations nanoparticle of enzyme sensitiveness as claimed in claim 1
Method, it is characterised in that in step 2), the molecular weight of the amphipathic polyester MePEG-Peptide-PER-CL is 22494.
6. the preparation of the amphipathic polyester MePEG-Peptide-PER-CL administrations nanoparticle of enzyme sensitiveness as claimed in claim 1
Method, it is characterised in that in step 2), the surfactant is dodecyl sodium sulfate.
7. the preparation of the amphipathic polyester MePEG-Peptide-PER-CL administrations nanoparticle of enzyme sensitiveness as claimed in claim 1
Method, it is characterised in that step
2) in, the mixing time is 4h.
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