CN102604076B - Multifunctional polyethylene glycol-dual vitamin E succinate derivative and application thereof in drug delivery - Google Patents
Multifunctional polyethylene glycol-dual vitamin E succinate derivative and application thereof in drug delivery Download PDFInfo
- Publication number
- CN102604076B CN102604076B CN 201210061032 CN201210061032A CN102604076B CN 102604076 B CN102604076 B CN 102604076B CN 201210061032 CN201210061032 CN 201210061032 CN 201210061032 A CN201210061032 A CN 201210061032A CN 102604076 B CN102604076 B CN 102604076B
- Authority
- CN
- China
- Prior art keywords
- succinate
- vitamin
- polyoxyethylene glycol
- drug
- methionin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Landscapes
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to a multifunctional polyethylene glycol-dual vitamin E succinate derivative and application thereof in drug delivery. An amphiphilic block copolymer utilizes polyethylene glycol as a hydrophyllic end to be combined with bimolecular hydrophobic vitamin E succinate under bridging of lysine, so that the AB2 type two-arm amphiphilic block copolymer is obtained. The polymer has potential antineoplastic activity and P-gp inhibitory action, and is capable of synergetically and effectively overcoming multidrug resistance to increase chemotherapeutic effect. Besides, the block copolymer can be self-assembled to form a micelle in aqueous medium, and can serve as a reservoir of poorly water-soluble drug, protein and gene-based drug, and the micelle is safe and high in stability and encapsulation, can be used for intravenous injection and oral ingestion, and has bright market application prospects.
Description
Technical field
The invention belongs to the new auxiliary material of pharmaceutical preparation and novel form field, relate to and a kind ofly have that antitumor action, P-gp suppress and the amphipathic ethylene glycol bisthioglycolate VE-succinate derivative of long circulatory function and the preparation of this carrier, and the application in useful for drug delivery as pharmaceutical carrier.
Background technology
For the clinical treatment of cancer, chemotherapy is the Main Means for the treatment of cancer.Yet, there is high toxicity, tumour non-exclusive and multidrug resistance due to chemotherapeutics and limited its clinical efficacy and application in application.Thereby the antitumor drug drug delivery system of development of new more and more draws attention.Amphipathic polymer micelle particularly causes concern in drug delivery system.Amphipathic nature block polymer self-assembly in water forms the nano-micelle of nucleocapsid structure, and insoluble drug (as taxanes, Anthraquinone) is wrapped in hydrophobic inner core, thereby has improved solvability and the stability of medicine.And for antitumor drug, drug-carrying polymer micelle can also reduce drug toxicity, suppress drug efflux and then overcome tumor multi-medicine drug-resistant, and polymer micelle can be gone up useful for drug delivery to tumor tissues by passive target by seepage and the retention effect (enhanced permeation and retention, EPR) of tumour largely.Thereby the parcel antitumor drug has great effect for overcoming multidrug resistance and increasing chemotherapeutic efficacy in nano-carrier.
Polyoxyethylene glycol (PEG) is a kind of low toxicity, the hydrophilic polymer of good biocompatibility, one of synthetic polymer that injection drug is used in the only a few of U.S. FDA approval can be as body.The Chang Zuowei hydrophilic group, can form the space multistory barrier, thereby can avoid the picked-up of reticuloendothelial system (RES), increases cycling time, and reach long circulating action, and then increase antitumor curative effect by the EPR effect.And VE-succinate has stronger hydrophobicity, and have anti-tumor activity and normal cell is not had to toxicity, the hydrophobic section using it as polymer micelle will have antitumor action synergistically.By both Chemical bond, make carrier there is the function of long circulation and tumor suppression, thereby increase the functional of micella.
At present, for fear of interior, take the photograph effluxing of medicine, overcome better the multidrug resistance of tumour, a lot of multidrug resistance inhibitor and Anticancer drug combination, but be limited to inefficiencies and high toxicity.Therefore develop a kind of new functional segmented copolymer, itself both can be used as the polymer support of micella, there is again long circulation, synergistic antitumor and suppress the multidrug resistance function, reduce toxicity and reach the drug combination purpose.This kind of segmented copolymer is applied to drug delivery system as carrier, will has great application prospect.
Summary of the invention
The object of the present invention is to provide a kind of long circulation that has, the antitumor amphipathic ethylene glycol bisthioglycolate VE-succinate derivative with suppressing the multidrug resistance function.
Second purpose of the present invention is to provide the preparation method of above-mentioned multi-functional amphipathic nature block polymer.
The 3rd purpose of the present invention is to provide the application of multi-functional amphipathic nature block polymer in pharmaceutical preparation.
The present invention is achieved through the following technical solutions above-mentioned purpose:
A kind of amphipathic multipolymer polyoxyethylene glycol two vitamin-E succinate derivatives, obtain good stability, functional strong, medicament solubilization carrier that encapsulation rate is high by polyoxyethylene glycol (PEG), Methionin and VE-succinate (VES) chemosynthesis, this polymer support is water-wet side by polyoxyethylene glycol, take Methionin as bridging, link hydrophobic VE-succinate.
The structural formula general formula of described amphipathic multipolymer polyoxyethylene glycol two vitamin-E succinate derivatives is as follows:
Described polymer support, is characterized in that the molecular weight ranges 2000-5000 of polyoxyethylene glycol used, can be one-ended hydroxy or amino a kind of or derivative.
A kind of preparation method of above-mentioned polyoxyethylene glycol two vitamin-E succinate derivatives, comprise the steps:
(a) VE-succinate is dissolved in methylene dichloride, in the 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride and I-hydroxybenzotriazole katalysis under, with benzyloxy Methionin in 30 ℃ of N
2the lower reaction of protection, obtain benzyloxy Methionin two VE-succinate, and structural formula is suc as formula I).
(b) get the product in step (a), under palladium carbon and hydrogen catalysis, obtain yellow solid Methionin two VE-succinate, structural formula is suc as formula II.
(c) step (b) product is in the 1-(3-dimethylaminopropyl)-catalysis of 3-ethyl-carbodiimide hydrochloride under, with the different chain length polyoxyethylene glycol, react, then ether recrystallization, drying, obtain faint yellow wax-like polyoxyethylene glycol two vitamin-E succinate derivatives (structural formula is shown in the formula III).
1-(3-dimethylaminopropyl described in step (a))-3-ethyl-carbodiimide hydrochloride and I-hydroxybenzotriazole composite catalyst catalysis VE-succinate are reacted with the bisamide of benzyloxy Methionin ester.
Described amphipathic ethylene glycol bisthioglycolate VE-succinate derivative has antitumor action, P-gp restraining effect and macrocyclic function.
Described amphipathic polyoxyethylene glycol two vitamin-E succinate derivatives, can be used as the insoluble drug solubilization carrier, wherein insoluble drug can be the arbitrary material or derivatives thereof in taxanes, camptothecin, Anthraquinones antitumour drug or dihydropyridines, NSAID (non-steroidal anti-inflammatory drug).
Described polymer micelle can adopt dialysis method, film dispersion method, and the emulsion process preparation method is characterized in that adopting following step:
Dialysis method is that amphipathic polyoxyethylene glycol two vitamin-E succinate derivatives and poorly water soluble drugs are dissolved in to organic solvent, in distilled water, dialyses, and 0.22 micron membrane filtration, obtain the drug-carrying polymer micelle of self-assembly.
Emulsion process is that above-mentioned polymkeric substance is soluble in water, under agitation slowly adds methylene dichloride or the chloroformic solution of medicine, and stirred overnight at room temperature is removed organic solvent, centrifugal, and 0.22 micron membrane filtration, obtain.
Film dispersion method is that above-mentioned polymkeric substance and medicine are dissolved in to organic solvent (as methyl alcohol, methylene dichloride etc.) simultaneously, and rotary evaporation, except organic solvent, obtains the film of medicine and carrier, add water to stir 2-5 h, centrifugal, 0.22 micron membrane filtration, obtain carrier micelle.
The present invention has following beneficial effect: prepare a kind of novel multi-functional amphipathic nature block polymer polyoxyethylene glycol two vitamin-E succinate derivatives, and carrier preparation process gentleness, easy to operate.Prepared drug-carrying polymer micelle, prepare easy, the less and homogeneous of particle diameter, encapsulation rate is high, micelle-forming concentration is low, good stability.Can be used as the bank of insoluble drug, protein or genomic medicine.In vitro cell experiment proves that amphipathic nature block polymer of the present invention is anticancer promotor and P-gp inhibitor, thereby, under its synergy, loads antitumor drug and can improve significantly chemotherapeutic efficacy.
The accompanying drawing explanation
The mono methoxy polyethylene glycol 2000-Methionin that Fig. 1 is the embodiment of the present invention 1-bis-VE-succinate block copolymer structures
1the HNMR spectrogram.
The Dynamic Light Scattering Determination micella grain-size graph of the drug-carrying polymer micelle that Fig. 2 is the embodiment of the present invention 6.
The perspective Electronic Speculum figure of the carrier micelle that Fig. 3 is the embodiment of the present invention 6.
Fig. 4 is embodiment 1 mono methoxy polyethylene glycol 2000-Methionin-bis-VE-succinate fluorescent exciting intensity rate I
335/ I
333with concentration logarithmic relationship figure.
Fig. 5 carries the release in vitro curve of Zorubicin mono methoxy polyethylene glycol 2000-Methionin-bis-VE-succinate micellas.
Fig. 6 is the graphic representation of the cell inhibitory rate for the MCF-7 cell that mtt assay is measured the segmented copolymer of the embodiment of the present invention 1.
The P-gp restraining effect that Fig. 7 is the cells were tested by flow cytometry segmented copolymer and the cellular uptake situation of carrier micelle.
Fig. 8 is curve during to the medicine after the mono methoxy polyethylene glycol 2000-Methionin of the quiet notes embodiment of rat 6 preparations-bis-VE-succinate Zorubicin carrier micelles and Zorubicin solution
Embodiment
Mode below by embodiment further illustrates the present invention, but therefore invention is not limited among described scope of embodiments.
The preparation of embodiment 1 mono methoxy polyethylene glycol 2000-Methionin-bis-VE-succinate segmented copolymers
(a) get 2.1 g VE-succinate and be dissolved in methylene dichloride, add the 1-(3-dimethylaminopropyl of 1 g)-I-hydroxybenzotriazole of 3-ethyl-carbodiimide hydrochloride and 0.6 g, stir 1 h at 0 ℃ of ice bath lower magnetic force.1.3 g benzyloxy Methionin ester hydrochloride sulfonate mixes with 3 mL triethylamines, then adds in above-mentioned activated solution N
2protect lower 30 ℃ of magnetic agitation 24 h.After completion of the reaction, washing, drying.Purifying, both obtained benzyloxy Methionin two VE-succinate (structural formula is suc as formula I).
(b) step (a) product is dissolved in to ethyl acetate, at palladium carbon (Pd/C) and H
2under effect, at 30 ℃ of magnetic agitation 6 h.Filter, collect filtrate, revolve steaming, obtain Methionin two VE-succinate (structural formula is suc as formula II).
(c) 1.2g step (b) is dissolved in methylene dichloride, add 0.44 g 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride and 0.13 g DMAP, 0 ℃ of lower magnetic force stirs 1 h activated carboxyl, then adds 2 g mono methoxy polyethylene glycols (MW=2000), N
2protect lower 37 ℃ of magnetic agitation 36 h.Filter, the ether recrystallization, collect white precipitate.Then obtain end product mono methoxy polyethylene glycol 2000-Methionin-bis-VE-succinate (structural formula is suc as formula III) in column chromatography for separation.Product is faint yellow wax.
Adopt nuclear magnetic resonance measuring
1hNMR hydrogen composes to determine the structure of segmented copolymer in embodiment 1, and the solvent of selecting is CDCl
3, result is as Fig. 1.5.7 ppm and 6.2 ppm are respectively two H in amido linkage-NH-CO-, 3.0ppm is following is the typical proton H peak in VE-succinate, and the proton peak of 3.7 ppm left and right is the H in PEG.
The preparation of embodiment 2 polyoxyethylene glycol 5000-Methionins-bis-VE-succinate segmented copolymers
(a) get 2.1 g VE-succinate and be dissolved in methylene dichloride, add the 1-(3-dimethylaminopropyl of 1 g)-I-hydroxybenzotriazole of 3-ethyl-carbodiimide hydrochloride and 0.6 g, stir 1 h at 0 ℃ of ice bath lower magnetic force.1.3 g benzyloxy Methionin ester hydrochloride sulfonate mixes with 3 mL triethylamines, then adds in above-mentioned activated solution N
2protect lower 30 ℃ of magnetic agitation 24 h.After completion of the reaction, washing, drying.Purifying, both obtained benzyloxy Methionin two VE-succinate (structural formula is suc as formula I).
(b) step (a) product is dissolved in to ethyl acetate, under palladium carbon and hydrogen effect, at 30 ℃ of magnetic agitation 6 h.Filter, collect filtrate, revolve steaming, obtain Methionin two VE-succinate (structural formula is suc as formula II).
(c) 1.2g step (b) is dissolved in methylene dichloride, add 0.44 g 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride and 0.13 g DMAP, 0 ℃ of lower magnetic force stirs 1 h activated carboxyl, then adds 5 g mono methoxy polyethylene glycols (MW=5000), N
2protect lower 37 ℃ of magnetic agitation 36 h.Filter, the ether recrystallization, collect white precipitate.Then obtain end product mono methoxy polyethylene glycol 5000-Methionin-bis-VE-succinate (structural formula is suc as formula III) in column chromatography for separation.Product is faint yellow wax.
In embodiment 1 and embodiment 2, selected polyoxyethylene glycol is monomethyl polyoxyethylene glycol (MW=2000,5000), but is not limited to this, and polyoxyethylene glycol of the present invention can also be mPEG-NH
2, NH
2-PEG-NH-Boc, MAL-PEG-NH
2and modify the polyethylene glycol hydroxyl, but be not limited to listed material.The molecular weight of polyoxyethylene glycol can be in the 2000-5000 scope.
The preparation of embodiment 3 mono methoxy polyethylene glycol amidation-Methionins-bis-VE-succinate segmented copolymers
(a) get 2.1 g VE-succinate and be dissolved in methylene dichloride, add the 1-(3-dimethylaminopropyl of 1 g)-I-hydroxybenzotriazole (HOBT) of 3-ethyl-carbodiimide hydrochloride (EDCI) and 0.6 g, stir 1 h at 0 ℃ of ice bath lower magnetic force.1.3 g benzyloxy Methionin ester hydrochloride sulfonate mixes with 3 mL triethylamines, then adds in above-mentioned activated solution N
2protect lower 30 ℃ of magnetic agitation 24 h.After completion of the reaction, washing, drying.Purifying, both obtained benzyloxy Methionin two VE-succinate (structural formula is suc as formula I).
(b) step (a) product is dissolved in to ethyl acetate, at palladium carbon (Pd/C) and H
2under effect, at 30 ℃ of magnetic agitation 6 h.Filter, collect filtrate, revolve steaming, obtain Methionin two VE-succinate (structural formula is suc as formula II).
(c) 1.2 g Lys-di-VES are dissolved in methylene dichloride, add 0.4 g EDCI and 0.2 g HOBT, and 0 ℃ of lower magnetic force stirs the 1h activated carboxyl, then add the amino (mPEG-NH of 1.6 g mono methoxy polyethylene glycols
2, MW=2000), N
2protect lower 37 ℃ of magnetic agitation 36 h.Filter, the ether recrystallization, collect faint yellow precipitation.Purifying obtains end product mono methoxy polyethylene glycol acid amides-Methionin-bis-VE-succinate (structural formula is suc as formula III).Product is faint yellow wax, and molecular weight is about 3152.
Amphipathic ethylene glycol bisthioglycolate VE-succinate derivative of the present invention can be used as the insoluble drug that carrier is applied to be had: taxol, Zorubicin, docetaxel, hydroxycamptothecine, camptothecine, vincristine(VCR), nimodipine, mitomycin etc.Especially to Zorubicin, docetaxel, taxol, solublization is arranged, but be not limited to listed medicine.
In the preparation of carrier micelle, the selection Zorubicin is model drug, embodiment 1 invention mono methoxy polyethylene glycol 2000-Methionin-bis-VE-succinate polymkeric substance are carrier.
The Zorubicin polymer micelle is carried in embodiment 4 dialysis method preparations
Take 5 mg Lipodoxs, 40mg mono methoxy polyethylene glycol 2000-Methionin-bis-VE-succinate, 10 μ L triethylamines, be dissolved in 4 mL DMFs, magnetic agitation 30 min make it even, solution is gone to (MWCO=1,4000) in dialysis tubing, as for dialysing in the 1L deionized water, respectively at 1 h, 2 h, 4 h, 6 h, 8 h, 12 h, 24 h change deionized water.After dialysis, take out the solution in dialysis tubing, centrifugal 20 min of 1,3000 rpm, remove the not medicine of parcel, 0.22 μ m membrane filtration, and filtrate is carrier micelle.The encapsulation rate of carrier micelle is 31.4%.
The Zorubicin polymer micelle is carried in embodiment 5 emulsification volatilization method preparations
Take 5 mg Lipodoxs, be dissolved in 3 mL methyl alcohol to 50 mL eggplant type bottles, add 10 μ L triethylamines, after stirring 0.5 h, 37 ℃ of rotary evaporation 30 min, remove organic solvent.Add 40 mg mono methoxy polyethylene glycol 2000-Methionins-bis-VE-succinate, be dissolved in 5 mL methylene dichloride, magnetic agitation is even.Dropwise add in the aqueous solution of 5 mL, 37 ℃ of stirrings are spent the night, and remove organic solvent.Centrifugal 20 min of 1,3000 rpm, remove not the medicine of parcel, 0.22 μ m membrane filtration and get final product.The encapsulation rate of carrier micelle is more than 80.2%, but still has residual organic solvent.
The Zorubicin polymer micelle is carried in embodiment 6 film dispersion method preparations
Take 5 mg Lipodoxs, be dissolved in 3 mL methyl alcohol to 50 mL eggplant type bottles, add 10 μ L triethylamines, after stirring 0.5 h, add 40 mg polymkeric substance mono methoxy polyethylene glycol 2000-Methionins-bis-VE-succinate, add 2ml methyl alcohol, jolting is even, 37 ℃ of rotary evaporation 30 min, remove organic solvent.Nitrogen gas stream (N
2) blow eggplant type bottle and remove residual organic solvent, add the aqueous solution of 5 mL, stir 2 h aquations in 37 ℃.Centrifugal 20 min of 1,3000 rpm, remove not the medicine of parcel, 0.22 μ m membrane filtration and get final product.The encapsulation rate of carrier micelle is more than 85.5%.
The carrier micelle of preparation in embodiment 6 is measured to size and the form of micella by dynamic light scattering and perspective Electronic Speculum, result is as Fig. 2 and Fig. 3.The quantity footpath of Dynamic Light Scattering Determination micella is 11.2 nm, and size distribution is narrower; Perspective Electronic Speculum figure shows that carrier micelle is the spherical of uniform particle diameter, and particle size range, at 10-15nm, conforms to the result of Dynamic Light Scattering Determination.
The mensuration of the amphipathic mono methoxy polyethylene glycol 2000-of test case 7 Methionin-bis-VE-succinate micelle-forming concentrations
The mensuration of micelle-forming concentration extensively adopts the fluorescence probe method.Pyrene is a kind of fat-soluble fluorescent probe, in the polarity environment fluorescence weak and in nonpolar environment fluorescence stronger.When micelle or hydrophobic region are arranged in polar solvent, pyrene can certainly be sent to nonpolar environment by the polarity environment and shift, and causes the fluorescence reinforcement.Usually this characteristic of pyrene is reacted by the ratio of the first excitation peak and the 3rd excitation peak intensity, when this ratio enlarges markedly, for pyrene is moved to nonpolar environment by the polarity environment, is micella or hydrophobic region and produces.
By 3 * 10
-6the ethanol solution of mol/L pyrene joins in the tool plug flask of 20 mL, and nitrogen gas stream dries up removes dehydrated alcohol.The polymkeric substance Mpeg2000-Lys-di-VES of embodiment 1 preparation of accurately weighed 10 mg is dissolved in the distilled water of 10 mL, obtains 1 mg/mL.Polymers soln is diluted to series concentration (10
-3, 10
-4, 10
-5, 10
-6, 10
-7, 10
-8, 10
-9mol/L) 10 mL, join respectively in the flask that contains pyrene, and the final concentration that makes pyrene is 3 * 10
-7mol/L, the ultrasonic 4h of lucifuge, lucifuge is placed and is spent the night.The fluorescent spectrophotometer assay fluorescence intensity.Emission wavelength is 390 nm, and excitation wavelength is I
1=333 nm, I
2=335 nm.
With I
335/ I
333fluorescence intensity ratio and the mapping of the concentration logarithm of polymer micelle as Fig. 4, point of inflexion on a curve is the micelle-forming concentration (CMC) of polymkeric substance, the CMC=1.14 that measurement result is mono methoxy polyethylene glycol 2000-Methionin-bis-VE-succinate.
Fig. 4 result proves that polymer support micelle-forming concentration of the present invention is lower, be easy to self-assembly and form micella, and the micella formed has dilution stability preferably.
The release in vitro of test case 8 carrier micelles
Adopt dialysis method to investigate mono methoxy polyethylene glycol 2000-Methionin-bis-VE-succinate and carry the tablets in vitro feature of Zorubicin micella.Pipette 1 mL1mg/mL polypeptide drug-loaded micelle solution in dialysis tubing, the dialysis tubing two ends clamp, be placed in respectively the Erlenmeyer flask that contains 30 mLpH7.4,6.5,5.5,4.5 PBS damping fluid release medium, in 37 ℃ of constant temperature oscillators, with 100 r/min, carry out the vitro release investigation.Sample 2 mL at 0.25,0.5,1,2,4,6,8,10,12,24 h respectively, supplement the release medium of respective volume simultaneously, dilution, by 0.45 μ m filtering with microporous membrane, is got 20 μ L and is carried out HPLC mensuration.Fig. 5 as a result.
As shown in Figure 5, the release of each carrier micelle has the pH sensitive features, and under physiological pH, release is comparatively slow, can make micella keep stable in working cycle, reaches long circulation then by EPR effect arrival target cell.And, under low pH environment, carrier micelle discharges the medicine of sealing in micella rapidly, prove in the target cell tissue and can bring into play rapidly curative effect.
By the human breast cancer cell (MCF-7) in logarithmic phase with 3 * 10
4the RPMI-1640 of/hole/0.1mL is embedded in 96 orifice plates, and the polymer support after 24h prepared by embodiment 1 is with 1,5,10,20,50 and 100 μ g/ml concentration add respectively each hole, every hole adds the polymer supported liquid solution of 100 μ L, and 3 parallel holes of each concentration, put in incubator and hatch 96h.After 96, take out 96 orifice plates, every hole adds the 2mg/mL MTT of 50 μ L, hatch 4h in incubator, knockout plate fully blots residual liquid by 96 orifice plate back-offs in filter paper, every hole adds the 200 μ L DMSO 10min that vibrates in vibrator, and microplate reader is measured the absorbancy that each hole 570nm goes out.Calculate inhibiting rate:
Inhibiting rate (%)=(1-A
medicine feeding hole/ A
control wells) * 100%
Mtt assay is measured polymkeric substance cytotoxicity result as Fig. 6, shows that block polymer mono methoxy polyethylene glycol 2000-Methionin-bis-VE-succinate have than strong cytotoxicity, can the kill tumor cell.Analyzing it is mainly the antitumor action due to VE-succinate.
Test case 10 P-gp suppress experiment
By the human breast cancer cell (MCF-7/Adr) of the adriamycin-resistant in logarithmic phase with 1 * 10
5the RPMI-1640 of/hole/2.5 mL is embedded in 6 orifice plates, after 24 h by Zorubicin solution, Zorubicin and 20 μ g/mL P-gp inhibitor cyclosporin A mixed solutions, the carrier micelle of the polymer support mixed solution of Zorubicin and 2 μ g/mL embodiment 1 preparations and example 5 preparations, be 6 μ g/mL concentration with the concentration of Zorubicin and add respectively each hole, put in incubator and hatch 2 h.Take out 6 orifice plates after 2 h, siphon away nutrient solution, PBS washing 3 times, trysinization, centrifugal, collecting cell, add 400 μ L PBS to mix, and adopts the cells were tested by flow cytometry sample.
The cells were tested by flow cytometry result is as shown in Figure 7: cyclosporin A is the inhibitor of P-gp, can increase the picked-up of P-gp substrate Zorubicin.And the mixed solution of Zorubicin and polymer support has increased the picked-up of Zorubicin, thereby prove that amphipathic nature block polymer of the present invention has certain P-gp restraining effect.The Zorubicin cellular uptake amount that also draws in embodiment 5 carrier micelle of preparation from figure obviously increases, and is mainly that to be wrapped in polymer latex intrafascicular due to (1) Zorubicin, can avoid the identification of P-gp, has increased the Zorubicin picked-up, thereby has overcome multidrug resistance; (2) work in coordination with in the P-gp restraining effect of the polymer support of embodiment 1 preparation, can suppress to enter effluxing of institute's free drug after cell, overcome synergistically multidrug resistance, increased the picked-up of P-gp medicine.
The pharmacokinetic of Zorubicin carrier micelle in the rat body that test case 11 mono methoxy polyethylene glycol 2000-Methionins-prepared by bis-VE-succinate
Get 12 health, male rat, body weight 250 g left and right, be divided into 2 groups at random, and fasting 12 h before administration, freely drink water.One group of intravenous injection Zorubicin solution (DOX) in contrast, another group is the carrier micelle of intravenous injection embodiment 5 preparations respectively, dosage is 5 mg/kg, 0.08,0.17,0.33,0.50,1.0,2.0,4.0,6.0,8.0,12,24,36,48,72,96,120,144,168,192,216,240h blood sampling time point after administration:, get blood 0.5 mL at each blood sampling time point by eye socket, after getting blood, move into immediately in the test tube of processing through heparin, centrifugal 10 min(13000 * g), 80 ℃ of refrigerator and cooled of separated plasma Yu – are frozen, and measure.During medicine, curve is shown in Fig. 8.
As shown in Figure 8, after giving the Zorubicin carrier micelle, with the solution group, compare, the Zorubicin micella is eliminated the transformation period significant prolongation, has long circulating action significantly.
Claims (7)
1. multi-functional polyoxyethylene glycol two vitamin-E succinate derivatives, it is characterized in that: this polymer support is water-wet side by polyoxyethylene glycol, take Methionin as bridging, links hydrophobic VE-succinate, structural formula is as follows:
2. polyoxyethylene glycol two vitamin-E succinate derivatives according to claim 1, is characterized in that, prepared derivative is AB
2type both arms segmented copolymer, the molecular weight polyethylene glycol scope is at 2000-5000.
3. the preparation method of polyoxyethylene glycol two vitamin-E succinate derivatives as claimed in claim 1, is characterized in that, adopts the following steps preparation:
(a) VE-succinate is dissolved in methylene dichloride, in the 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride and I-hydroxybenzotriazole katalysis under, with benzyloxy Methionin in 30 ℃ of N
2the lower reaction of protection, obtain benzyloxy Methionin two VE-succinate I;
(b) get the product in step (a), under palladium carbon and hydrogen catalysis, obtain yellow solid Methionin two VE-succinate II;
(c) step (b) product is in the 1-(3-dimethylaminopropyl)-catalysis of 3-ethyl-carbodiimide hydrochloride under, with the different chain length polyoxyethylene glycol, react, then ether recrystallization, drying, obtain faint yellow wax-like polyoxyethylene glycol two vitamin-E succinate derivative III
The described polyoxyethylene glycol two vitamin-E succinate derivatives of claim 1 or 2 antitumor in preparation, suppress the application in the P-gp medicine.
5. the application of the described polyoxyethylene glycol two vitamin-E succinate derivatives of claim 1 or 2 in the long circulation medicine of preparation.
6. polyoxyethylene glycol two vitamin-E succinate derivatives according to claim 1 and 2, it is characterized in that, described derivative, by dialysis method, film dispersion method or emulsion process, forms drug-carrying polymer micelle with hydrophobic drug, protein, the self-assembly of gene class medicine.
7. polyoxyethylene glycol two vitamin-E succinate derivatives according to claim 6, it is characterized in that: described hydrophobic drug is the arbitrary material or derivatives thereof in taxanes, camptothecin, Anthraquinones antitumour drug or dihydropyridines, NSAID (non-steroidal anti-inflammatory drug).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201210061032 CN102604076B (en) | 2012-03-09 | 2012-03-09 | Multifunctional polyethylene glycol-dual vitamin E succinate derivative and application thereof in drug delivery |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201210061032 CN102604076B (en) | 2012-03-09 | 2012-03-09 | Multifunctional polyethylene glycol-dual vitamin E succinate derivative and application thereof in drug delivery |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102604076A CN102604076A (en) | 2012-07-25 |
| CN102604076B true CN102604076B (en) | 2013-12-25 |
Family
ID=46521888
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201210061032 Expired - Fee Related CN102604076B (en) | 2012-03-09 | 2012-03-09 | Multifunctional polyethylene glycol-dual vitamin E succinate derivative and application thereof in drug delivery |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102604076B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112006986A (en) * | 2020-08-07 | 2020-12-01 | 厦门大学 | Vitamin E succinate polyethylene glycol nano micelle and preparation method and application thereof |
Families Citing this family (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104116709A (en) * | 2013-04-23 | 2014-10-29 | 北京大学 | Tumor-targeting pH-sensitive polymeric micelle composition resisting tumor drug resistance |
| CN104116711A (en) * | 2013-04-23 | 2014-10-29 | 北京大学 | pH-sensitive polymeric micelle composition resisting tumor drug resistance |
| CN103788366B (en) * | 2014-01-22 | 2015-10-28 | 沈阳药科大学 | Mono methoxy polyethylene glycol-two sulphur-two VE-succinate and Synthesis and applications thereof |
| CN104356380B (en) * | 2014-10-23 | 2016-06-29 | 浙江工业大学 | Epsilon-polylysine-vitamin e succinate amide complex and preparation method thereof and application |
| CN104399084B (en) * | 2014-11-29 | 2017-08-25 | 沈阳药科大学 | The application of PEGylation vitamin A acid and its self-assembled micelle in drug delivery |
| CN104549084B (en) * | 2014-12-04 | 2016-06-22 | 浙江工业大学 | Composite micro-capsule wall material and microcapsule and preparation method |
| CN104530256B (en) * | 2014-12-18 | 2017-04-26 | 北京中医药大学 | Hyaluronic acid-vitamin E succinate polymer as well as preparation and application thereof |
| CN104672441B (en) * | 2015-02-10 | 2016-05-11 | 长春艾诺生化医药有限责任公司 | Polyethylene glycol-sulfenyl is sad-vitamin E copolymer and preparation method and application |
| CN105381468A (en) * | 2015-11-18 | 2016-03-09 | 沈阳药科大学 | Preparation of PLV2K-PLGA nano-particles with P-gp inhibiting effect and application of PLV2K-PLGA nano-particles in oral administration |
| CN105726511A (en) * | 2016-03-10 | 2016-07-06 | 温州医科大学 | Self-assembled polylysine graft nanoparticles and preparation and application thereof |
| CN105853357A (en) * | 2016-04-26 | 2016-08-17 | 中国药科大学 | Multifunctional cooperative mixed micelle drug delivery system and preparation method thereof |
| CN107674196B (en) * | 2017-10-23 | 2020-01-10 | 天津工业大学 | Docetaxel prodrug with anti-tumor effect and preparation method thereof |
| CN108670954B (en) * | 2018-06-12 | 2021-04-20 | 宁夏医科大学 | Chemotherapeutic drug co-loaded glycyrrhetinic acid prodrug micelle and preparation method thereof |
| CN109125260B (en) * | 2018-07-30 | 2020-10-02 | 北京中医药大学 | Application of amphiphilic polymer in preparation of oral medicine |
| CN112121010A (en) * | 2020-09-02 | 2020-12-25 | 广州市第一人民医院（广州消化疾病中心、广州医科大学附属市一人民医院、华南理工大学附属第二医院） | Reduction response type vitamin E polyethylene glycol succinate micelle containing folic acid molecules and preparation method and application thereof |
| CN114181200B (en) * | 2021-11-24 | 2023-09-29 | 华中科技大学 | Cationic liposome with efficient gene transfection efficiency and preparation and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101468203A (en) * | 2007-12-25 | 2009-07-01 | 沈阳药科大学 | Method for preparing cleavable polyethyleneglycol lipid derivates and application |
| CN102068701A (en) * | 2011-01-18 | 2011-05-25 | 沈阳药科大学 | Application of cleavable polyethylene glycol (PEG) lipid derivative to preparation |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DK2229156T3 (en) * | 2007-12-20 | 2017-02-06 | Fertin Pharma As | CHEW GUM TABLE AND PROCEDURE FOR DOSING PHARMACEUTICAL ACTIVE INGREDIENTS IN SUCH CHEW GUM TABLE |
-
2012
- 2012-03-09 CN CN 201210061032 patent/CN102604076B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101468203A (en) * | 2007-12-25 | 2009-07-01 | 沈阳药科大学 | Method for preparing cleavable polyethyleneglycol lipid derivates and application |
| CN102068701A (en) * | 2011-01-18 | 2011-05-25 | 沈阳药科大学 | Application of cleavable polyethylene glycol (PEG) lipid derivative to preparation |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112006986A (en) * | 2020-08-07 | 2020-12-01 | 厦门大学 | Vitamin E succinate polyethylene glycol nano micelle and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102604076A (en) | 2012-07-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102604076B (en) | Multifunctional polyethylene glycol-dual vitamin E succinate derivative and application thereof in drug delivery | |
| Zhang et al. | Folate-decorated poly (lactide-co-glycolide)-vitamin E TPGS nanoparticles for targeted drug delivery | |
| Zhu et al. | Docetaxel (DTX)-loaded polydopamine-modified TPGS-PLA nanoparticles as a targeted drug delivery system for the treatment of liver cancer | |
| Zhao et al. | Redox-sensitive mPEG-SS-PTX/TPGS mixed micelles: an efficient drug delivery system for overcoming multidrug resistance | |
| Pan et al. | Targeted delivery of paclitaxel using folate-decorated poly (lactide)–vitamin E TPGS nanoparticles | |
| Zhang et al. | A pH-sensitive nanosystem based on carboxymethyl chitosan for tumor-targeted delivery of daunorubicin | |
| Wang et al. | Star-shape copolymer of lysine-linked di-tocopherol polyethylene glycol 2000 succinate for doxorubicin delivery with reversal of multidrug resistance | |
| Zhang et al. | Nucleolin targeting AS1411 aptamer modified pH-sensitive micelles for enhanced delivery and antitumor efficacy of paclitaxel | |
| CN102626518B (en) | Preparation and application of insoluble drug-entrapped poloxamer/amphiphilic polysaccharide mixed micelle | |
| Kulhari et al. | Cyclic-RGDfK peptide conjugated succinoyl-TPGS nanomicelles for targeted delivery of docetaxel to integrin receptor over-expressing angiogenic tumours | |
| CN102060991B (en) | Amphiphilic prodrug of 7- ethyl-10-hydroxycamptothecin and preparation method thereof | |
| Pu et al. | A polymeric micelle with an endosomal pH-sensitivity for intracellular delivery and enhanced antitumor efficacy of hydroxycamptothecin | |
| CN101991860B (en) | Poloxamer-carboxylic acid drug conjugate and preparation method and application thereof | |
| Shi et al. | Docetaxel-loaded PEO–PPO–PCL/TPGS mixed micelles for overcoming multidrug resistance and enhancing antitumor efficacy | |
| CN101721350B (en) | Folate-mediated targeted polymeric micelle | |
| Qin et al. | mPEGylated solanesol micelles as redox-responsive nanocarriers with synergistic anticancer effect | |
| Lei et al. | Co-delivery of paclitaxel and gemcitabine via a self-assembling nanoparticle for targeted treatment of breast cancer | |
| Zhang et al. | Design and evaluation of a PEGylated lipopeptide equipped with drug-interactive motifs as an improved drug carrier | |
| Liu et al. | Self-assembled nanoparticles based on a carboxymethylcellulose–ursolic acid conjugate for anticancer combination therapy | |
| Wang et al. | Preparation and properties of hydroxycamptothecin-loaded nanoparticles made of amphiphilic copolymer and normal polymer | |
| CN105188673A (en) | Formulations and carrier systems including compound interactive domains | |
| CN106883404B (en) | Polyethylene glycol vitamin E succinate derivative and its preparation method and application | |
| CN103788366B (en) | Mono methoxy polyethylene glycol-two sulphur-two VE-succinate and Synthesis and applications thereof | |
| Li et al. | Sorafenib-loaded nanoparticles based on biodegradable dendritic polymers for enhanced therapy of hepatocellular carcinoma | |
| Li et al. | Active targeting co-delivery system based on pH-sensitive methoxy-poly (ethylene glycol) 2K-poly (ε-caprolactone) 4K-poly (glutamic acid) 1K for enhanced cancer therapy |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20131225 Termination date: 20190309 |