CN103977434A - P-hydroxybenzoic acid mediated polymer micelle drug delivering system with brain targeting function - Google Patents
P-hydroxybenzoic acid mediated polymer micelle drug delivering system with brain targeting function Download PDFInfo
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Abstract
The invention belongs to the technical field of high polymer material and pharmaceutical preparations, and particular to preparation of a p-hydroxybenzoic acid mediated polymer micelle drug delivering system with brain targeting function and application of the polymer micelle drug delivering system in the treatment of brain tumors. The polymer micelle drug delivering system with the brain targeting function is prepared by mixed construction of a brain targeting functional material and an amphipathic r polymer material, in-vivo and in-vitro activity evaluation results show that the p-hydroxybenzoic acid mediated polymer micelle drug delivering system can step over blood brain barrier to deliver drugs into the brain, can significantly increase the drug accumulation in the brain, and has the obvious antitumor effect, and compared with a common polymer micelle drug delivery system, the treatment effect of anticancer drugs on the brain tumor can be significantly improved.
Description
Technical field
The invention belongs to macromolecular material and technical field of medicine, be specifically related to polymer micelle delivery system of a kind of P-hydroxybenzoic acid mediated brain targeting and preparation method thereof, and the application of this brain target polymer micelle delivery system in brain tumor treatment.
Background technology
It is the common malignant disease of central nervous system that prior art discloses brain tumor, comprise brain primary tumo(u)r and brain metastatic tumour, because tumor locus approaches patient central nervous system, clinical general operation is difficult to remove completely and tumor easily recurs, therefore, in clinical practice, conventionally adopt the control for tumor recurrence in conjunction with chemotherapy of excision brain tumor.
In prior art, targeting drug delivery system for brain tumor treatment adopts brain targeting or cancer target nanoscale medicine delivery system more, such targeted nano delivery system normally utilizes the receptor of brain capillary endothelial cell specifically expressing, or brain tumor organizes specific recognition and the combination of receptor with the corresponding part on nanoscale medicine delivery system of new vessels endotheliocyte or tumor cell specifically expressing, to realize the targeted delivery of antitumor drug.
Studies show that, blood-brain barrier (BBB) is a kind of adjustment interface between the neuronal cell of blood and brain, spinal cord, and mass exchange between central nervous system (CNS) and peripheral blood is played regulatory role.BBB has three-decker: internal layer be brain capillary endothelial cell (BCECs) and between tight connection, centre is basement membrane and pericyte, skin is astrocyte and extracellular matrix, and wherein BCECs and tight connection thereof are the principal elements that forms BBB barrier.Due to the existence of BBB and cerebrospinal fluid barrier (BCSFB), make the treatment of nearly all macromolecular drug and 98%, diagnosis brain diseases medicine cannot enter brain and central nervous system.Wherein, except fat-soluble, the relative molecular mass of medicine itself and ability of formation hydrogen bond etc. are to cause medicine to be difficult to the reason by BBB, BBB also has efflux mechanism, by the P-glycopeptide on BBB, some medicines is transported in brain.In addition; " the enzymology blood brain barrier " of the upper existence of BBB; it is highly active neuropeptide digestive enzyme; as; the amine peptidase of being combined with blood capillary, endopeptidase, angiotensin converting enzyme (ACE) etc., also making affects the treatment to brain diseases with the medicine of peptide coupling because metabolism is unstable, therefore; described BBB had both effectively protected cerebral tissue, simultaneously also for Drug therapy brain diseases has been manufactured the barrier that is difficult to go beyond.
At present, about the method that overcomes BBB increase medicine brain delivery is divided into invasive and the large class of Noninvasive two by administering mode.Invasive medication comprises that height oozes shock, carotid injection vaso-active substance and directly intracerebral injection administration.Although the method is effective, may cause damage and the surgery damage of brain infection, BBB, and administering mode complexity, patient compliance is poor, is not suitable for as conventional treatment and diagnosis scheme.Comparatively speaking, Noninvasive medication has more wide potential applicability in clinical practice, and the medication of Noninvasive mainly comprises: the structural modification of medicine, chemical transmission system, carrier mediated transhipment, transcytosis and nasal drug delivery system; First three methods all needs medicine to carry out certain chemical modification, and this chemical constitution, physicochemical property etc. for medicine is had relatively high expectations, and has larger limitation.
Benzamide compound is d2 dopamine receptor selective antagonist, has higher specificity and stronger affinity, and the side reaction of its extrapyramidal system is very little.It is reported, this type of medicine and the d2 dopamine receptor structure activity relationship on molecular level is: pyrrole ring N atom is one of this compounds and the interactional site of d2 dopamine receptor, this N atomic charge is larger, more be conducive to the COO-Interaction of substituents on compound and acceptor amino acid residue, stronger with the affinity of d2 dopamine receptor; Phenyl ring is interactional another site of benzamide compound and d2 dopamine receptor, phenyl ring is combined with the hydrophobic parts of acceptor molecule, its in conjunction with situation be subject to phenyl ring plane electrically charged and space structure phenyl ring affect, the charge distributing value of phenyl ring plane is less, phenyl ring and amide group keep coplanar space structure simultaneously, and the affinity of compound and d2 dopamine receptor is stronger.Therefore, the work that at present home and abroad researcher does is mainly to carry out structural modification and transformation for group and benzene ring side chain on the pyrrole ring N atom of this compounds, and successively synthetic and studied a series of derivants.
The present invention intends utilizing benzamide compound to have the feature of affine d2 dopamine receptor, prepares a kind of brain target polymer micelle delivery system of P-hydroxybenzoic acid mediation.Up to now, there is not yet the preparation of brain target polymer micelle delivery system and the bibliographical information of application in brain tumor treatment thereof of the mediation of relevant P-hydroxybenzoic acid.
Summary of the invention
The object of the present invention is to provide a kind of new brain targeting drug delivery system, be specifically related to a kind of brain target polymer micelle delivery system of P-hydroxybenzoic acid mediation.The present invention has built one can penetrate blood brain barrier, realize the polymer micelle delivery system of the P-hydroxybenzoic acid modification of brain drug delivery, this delivery system can be delivered to chemotherapeutics, diagnostic medicine in brain by intravenously administrable approach, performance treatment and diagnosis effect.
The brain target polymer micelle delivery system of P-hydroxybenzoic acid mediation of the present invention, comprise targeted molecular, carrier and medicine, described targeted molecular is P-hydroxybenzoic acid, described carrier is the amphipathic nature block polymer polymer micelle with nucleocapsid structure of spontaneous formation under proper condition, carry or suction type medicine carrying by bag, can obviously improve the enter brain volume of medicine through blood brain barrier.
It is brain targeted molecular that the present invention adopts P-hydroxybenzoic acid.
In the present invention, carrier material is selected amphipathic nature polyalcohol material, and its hydrophobic fragment is PHOSPHATIDYL ETHANOLAMINE, and hydrophilic segment is the Polyethylene Glycol of one end methoxyl group end-blocking, and its molecular weight is 1000-20000 Da, preferred molecular weight 2000-5000 Da.
In delivery system of the present invention, its medicine carrying mode is parcel or Electrostatic Absorption.
More specifically, the brain target polymer micelle delivery system of P-hydroxybenzoic acid mediation of the present invention, it is characterized in that, be jointly built into by thering is brain target function material (I), amphipathic nature polyalcohol material (II) and medicine the polymer micelle delivery system that wraps medicine carrying thing; Brain target function material (I) accounts for the 2%-20% of amphipathic nature polyalcohol material (II); Wherein: brain target function material (I) is made up of the mPEG2000-DSPE of modifying through P-hydroxybenzoic acid; Amphipathic nature polyalcohol material (II) is made up of the mPEG2000-DSPE of methoxyl group end-blocking.In embodiments of the invention, the molecular weight polyethylene glycol in brain target function material is between 2000-5000; Molecular weight polyethylene glycol in amphipathic nature polyalcohol material is between 1000-5000.
In the present invention, the PHOSPHATIDYL ETHANOLAMINE in brain target function material or amphipathic nature polyalcohol material, is satisfied fatty acid PHOSPHATIDYL ETHANOLAMINE or unsaturated fatty acid PHOSPHATIDYL ETHANOLAMINE.
In the present invention, the medicine that delivery system is sent is one or more in diagnostic medicine or medicine;
Wherein, diagnostic medicine comprises: nuclear medicine diagnostic medicine is as radiosiotope technetium or indium, and MR diagnosis medicine is as metal ion gadolinium; Medicine comprises: lipotropy antitumor drug, is selected from paclitaxel (PTX), Docetaxel (DTX), amycin (DOX), camptothecine, hydroxy camptothecin, 9-nitrocamptothecin, levodopa (L-DOPA) etc.
In the present invention, the particle diameter of described polymer micelle delivery system is 7-30nm.
The invention provides the preparation method of brain target polymer micelle delivery system of described P-hydroxybenzoic acid mediation and the application in brain tumor treatment thereof.
The present invention adopts thin film aquation legal system for polymer micelle delivery system, wherein: respectively brain target function material (I), amphipathic nature polyalcohol material (II) and medicine are dissolved in to organic solvent altogether, after film forming, add aquation medium (as: normal saline, phosphate buffer, 5% glucose solution) aquation to form polymer micelle; Its midbrain target function material (I) is mol ratio 5%, 10%, 15% or 20% with the ratio of amphipathic nature polyalcohol material (II).
Object of the present invention is achieved through the following technical solutions:
1, the fluorescent labeling of P-hydroxybenzoic acid-polymer micelle
P-hydroxybenzoic acid is placed under room temperature and is reacted with amino PEG2000-DSPE, and product is by dialysis method purification process;
With brain target function material and framework material mol ratio 5:95, utilize the standby bag of film forming aquation legal system to carry the polymer micelle of Coumarin-6 (C6) and DiR, remove free fluorescein by gel chromatography;
2, the preparation of P-hydroxybenzoic acid-polymer micelle/DTX
With brain target function material and framework material mol ratio 5:95, utilize the standby bag of film forming aquation legal system to carry the polymer micelle of chemicals, in one embodiment of the present of invention, chemicals is selected from Docetaxel (DTX);
3, the picked-up of BCECs cell test
Utilize fluorescence microscope and flow cytometer, measure the qualitative, quantitative picked-up situation of the polymer micelle delivery system of brain capillary endothelial cell BCECs to bag year C6;
4, animal tissue's distribution test
Adopt small animal living body imaging system, the distribution situation of the delivery system of observation DiR labelling in normal mouse body, and observe fluorescence distribution situation in each internal organs after processing mice;
5, the effect of P-hydroxybenzoic acid-polymer micelle/DTX is evaluated
Carry out mtt assay and measure the growth inhibited Experiment on Function of polymer micelle to brain glioblastoma cell U87 MG on cellular level, result demonstration, the polymer micelle delivery system making can significantly improve the inhibitory action of medicine to growth of tumour cell; Original position Brain Glioma Model Mus pharmacodynamics test shows, the brain target polymer micelle delivery system of P-hydroxybenzoic acid of the present invention mediation has good blood brain barrier and the brain congregational rate of penetrating, and can significantly improve the life span of Glioma Model animal.
The present invention is by Coumarin-6 and nir dye DiR fluorescent labeling tracer technique, and the polymer micelle delivery system of P-hydroxybenzoic acid mediated brain targeting carries out external BCECs cellular uptake and mice live body, in vitro brain targeting characterize test; Carry slightly solubility chemicals Docetaxel by P-hydroxybenzoic acid-polymer micelle bag, carry out the inside and outside anti-glioma pharmacodynamic evaluation of Mice Body, result shows, this brain target polymer micelle delivery system can be realized the target that improves brain tumor targeted therapy effect, reduces toxic and side effects; Described brain target polymer micelle delivery system has single-minded brain targeting, can overcome BBB and special antitumor drug (as: paclitaxel, the Docetaxel sent, the lipophilic drugs such as camptothecine) arrive brain, thus there is characteristic efficient, the anti-brain tumor of low toxicity.
The present invention passes through internal and external test, result confirms, described brain target polymer micelle delivery system has brain target function simultaneously, can carry medicine through BBB and targeted delivery of drugs to brain, strengthen the distribution of fluorescent probe at brain, and make the antitumor drug that bag carries there is good anti-cerebral glioma therapeutic effect, especially reach special brain Targeting Effect.This polymer micelle delivery system has a good application prospect for diseases such as treatment brain tumors.
tool of the present invention has the following advantages:
1, novel brain target function material is provided, be conducive to improve polymer micelle leap BBB and enter the ability in brain, with protein polypeptide brain targeted molecular comparisons such as the monoclonal antibody with brain target function of prior art, transferrins, lactoferrin, cation protein, RVG, p-hydroxybenzoic acid amount of the present invention is little, non-immunogenicity, preparation method are simple, cheap and easy to get;
2, provide the functional material of brain target polymer micelle delivery system to prepare and compound mode, preparation method is simple, is convenient to subsequent development and industrialization;
3, brain target polymer micelle delivery system is conducive to improve the medicine degree that concentrates in brain, improves the therapeutic effect of medicine to diseases such as brain tumors.
brief description of the drawings:
fig. 1: p-HA-PEG-DSPE's
1
h-NMR collection of illustrative plates
Wherein show that synthetic product has between 0.8-1.5ppm
two, there is the characteristic peak of Polyethylene Glycol in the characteristic peak of stearoyl PHOSPHATIDYL ETHANOLAMINE, occurred the characteristic peak of P-hydroxybenzoic acid at 6.8-7.8ppm between 3.2-4.0ppm, and p-HA-PEG-DSPE is confirmed.
fig. 2: the qualitative picked-up test of BCECs cell to P-hydroxybenzoic acid-polymer micelle (p-HA-micelles)
Wherein, A, B are light field and the details in a play not acted out on stage, but told through dialogues of free C6 group, and C, D are light field and the details in a play not acted out on stage, but told through dialogues of mPEG-micelles/C6 group, and E, F are light field and the details in a play not acted out on stage, but told through dialogues of p-HA-micelles/C6 group; After administration 1 h, free C6, mPEG-micelles/C6 and p-HA-micelles/C6 all can enter cell, but the cell fluorescence intensity of p-HA-micelles/C6 micelle group is higher than free C6 group and mPEG-micelles/C6 group, and the micelle of modifying through p-HA can increase the picked-up of BCECs cell to C6.
fig. 3: the quantitative picked-up test of BCECs cell to P-hydroxybenzoic acid-polymer micelle (p-HA-micelles)
Wherein, A is free C6 group, and B is mPEG-micelles/C6 group, and C is p-HA-micelles/C6 group; After administration 1 h, free C6, mPEG-micelles/C6 and p-HA-micelles/C6 all can enter cell, its uptake ratio be respectively 75%, 79.42% and the cell fluorescence intensity of 92.88%, p-HA-micelles/C6 group be significantly higher than free C6 group and mPEG-micelles/C6 group (
p< 0.05), p-HA micelle can increase the picked-up of BCECs cell to C6, and the micelle of unmodified is without this advantage.
fig. 4: the mice biological tissue scattergram of P-hydroxybenzoic acid-polymer micelle
Wherein show that mouse tail vein injects respectively after mPEG-micelles/DiR, p-HA-micelles/DiR 24 hours, living body fluorescent image after 10% chloral hydrate anesthesia, the mice of one, the left side for injection mPEG-micelles/DiR, the mice of one, the right for injection p-HA-micelles/DiR; Can be found out by fluorescence distribution figure, the fluorescence distribution of p-HA-micelles/DiR in mouse brain is apparently higher than mPEG-micelles/DiR, and prompting p-HA-micelles/DiR can penetrate BBB by the mediation of P-hydroxybenzoic acid targeting and enter brain.
fig. 5: the mice in vitro tissue scattergram of P-hydroxybenzoic acid-polymer micelle,
Wherein show that mouse tail vein injects respectively different time points after mPEG-micelles/DiR and p-HA-micelles/DiR, 10% chloral hydrate anesthesia, the in vitro each internal organs fluorescence distribution figure of mice and semidefinite spirogram after normal saline cardiac perfusion, fluorescence distribution figure by 2 hours, 4 hours, 8 hours, 12 hours finds out, p-HA-micelles/DiR each time point in mouse brain has obvious fluorescence distribution, and prompting p-HA-micelles can be mediated and penetrated BBB and distribute in brain by P-hydroxybenzoic acid.
fig. 6: original position glioma model Mus median survival time figure
P-HA-micelles/DTX group, mPEG-micelles/DTX group, Taxotere are wherein shown
?the median survival time of group and normal saline (physiological saline) group is respectively 45.8,32,27 and 22 days, the median survival time that p-HA-micelles/DTX can significant prolongation original position glioma model Mus is described, there is good anti-glioma effect.
Detailed description of the invention
Describe and will contribute to further to understand the present invention by following examples, but the present invention is not limited to describe as follows scope.
embodiment 1 prepares P-hydroxybenzoic acid-polymer micelle/DTX(p-HA-micelles/DTX)
1) prepare P-hydroxybenzoic acid-PEG2000-DSPE (p-HA-PEG-DSPE)
Take appropriate p-HA, be dissolved in DMF, add a certain amount of NHS, EDCHCl to activate 3 h, centrifuging and taking supernatant; Take appropriate NH
2-PEG-DSPE, in DMF, dropwise adds supernatant, then adds the DIEA of 50 μ L, under room temperature, stirs and spends the night.With ice ether to above-mentioned reactant liquor precipitate, centrifugal.After vacuum drying, disperse dry thing with distilled water, dialysis purification, lyophilization can obtain white solid p-HA-PEG-DSPE.Adopt
1h-NMR verifies (500 MHz, CD to its structure
3cl) (as shown in Figure 1), result shows: this compound has between 0.8-1.5ppm
two, there is the characteristic peak of Polyethylene Glycol in the characteristic peak of stearoyl PHOSPHATIDYL ETHANOLAMINE, occurred the characteristic peak of P-hydroxybenzoic acid at 6.8-7.8ppm between 3.2-4.0ppm, confirms that the product that obtains is p-HA-PEG-DSPE.
) prepare and wrap the polymer micelle that carries DTX
Take appropriate Docetaxel (DTX) and mPEG-DSPE, be dissolved in acetonitrile, both vortexs are mixed.Above-mentioned solution is put on Rotary Evaporators, 37 DEG C of decompression aquation film forming 2 h, adopt 3 mL aquation media to carry out aquation, by 0.22 μ m filtering with microporous membrane for resulting polymers micelle delivery system, obtain the DTX polymer micelle (mPEG-micelles/DTX) of unmodified;
Adopt above-mentioned identical preparation technology, and in prescription, add 5%(p-HA-PEG-DSPE/mPEG-DSPE mol ratio) p-HA-PEG-DSPE, preparation is loaded with the polymer micelle (p-HA-micelles/DTX) that the P-hydroxybenzoic acid (p-HA) of DTX is modified.
the BCECs cellular uptake test of embodiment 2 P-hydroxybenzoic acid-polymer micelle (p-HA-micelles)
1) prepare P-hydroxybenzoic acid-polymer micelle/C6(p-HA-micelles/C6)
Adopt film forming aquation method, prepare the mPEG-micelles of Coumarin-6 (C6) labelling, specifically comprise step: mPEG-DSPE is mixed with 50 μ g fluorescent dye C6, vortex 1 min, in 37 DEG C of rotation evaporate to dryness film forming, vacuum drying is removed residual organic solvent, add HEPES(pH 7.4,10 mmol/L) 200 μ L, vortex 10 min, 37 DEG C of water-bath jolting 2 h, adopt gel filtration chromatography, taking G50 as filler, HEPES is the C6 that eluting medium is removed unentrapped, cross after the microporous filter membrane of 0.22 μ m, obtain mPEG-micelles/C6; Adopt identical preparation technology, with p-HA-PEG-DSPE and mPEG-DSPE mol ratio 5:95, be prepared into p-HA-micelles/C6, with fluorescent spectrophotometer assay C6 content, calculate content by standard curve.
2) the qualitative picked-up test of the BCECs cell of P-hydroxybenzoic acid-polymer micelle (p-HA-micelles)
The take the logarithm BCECs cell of trophophase, with 0.25 % trypsinization and blow and beat into individual cells, cell suspension in the DMEM culture fluid containing containing 10%FBS and 1% penicillin-streptomycin, counting, be inoculated in 24 orifice plates with 20000, each hole cell, every pore volume 1 mL, cultivates in CO2 gas incubator after 24 h, washes 2 times with the PBS of pH 7.4.Add free C6, mPEG-micelles/C6 and each 1 mL of p-HA-micelles/C6, wherein the concentration of C6 is 0.05 μ g/mL, after hatching 1 h in carbonoxide case, stop, PBS washing 3 times, cell is fixed 15 min with 10% paraformaldehyde solution, PBS washing 3 times, with DAPI(1 μ g/mL) dye core 10 min, again with PBS washing 3 times, finally with buffering glycerol mounting, be placed in fluorescence microscopy Microscopic observation, result shows, BCECs to the intake of p-HA-micelles/C6 apparently higher than free C6 and mPEG-micelles/C6(as shown in Figure 2).
3) the BCECs cell of P-hydroxybenzoic acid-polymer micelle (p-HA-micelles/) quantitatively absorbs test
The take the logarithm BCECs cell of trophophase, with 0.25 % trypsinization and blow and beat into individual cells, cell suspension in the DMEM culture fluid containing containing 10%FBS and 1% penicillin-streptomycin, counting, be inoculated in 24 orifice plates with 20000, each hole cell, every pore volume 1 mL, in CO2 gas incubator, cultivate after 24 h, wash 2 times with the PBS of pH 7.4, add free C6, each 1 mL of mPEG-micelles/C6 and p-HA-micelles/C6, wherein the concentration of C6 is 25 ng/mL, after hatching 1 h in carbonoxide case, stop, PBS washing 3 times, after adding appropriate pancreatin that cell dissociation is centrifugal, abandoning supernatant, with the PBS washed cell of pH 7.4 2 times, and finally make cell be suspended in the PBS of 0.5 mL, detect fluorescence intensity proportion in each time point tumor cell with flow cytometer, result shows, BCECs to the intake of p-HA-micelles/C6 apparently higher than free C6 and mPEG-micelles/C6(as shown in Figure 3)
the brain drug delivery animal experiment of embodiment 3 P-hydroxybenzoic acid-polymer micelle (p-HA-micelles)
1) the mice biological tissue of P-hydroxybenzoic acid-polymer micelle/DiR distributes
Prepare respectively p-HA-micelles/DiR
, mpEG-micelles/DiR polymer micelle, wherein the consumption of DiR is 15.6 μ g, mouse tail vein injection 100 μ L/ only, respectively at after administration 1,2,4,8,12 and 24 hour with 10% chloral hydrate anesthesia mice, in living animal imaging system, observe fluorescence distribution in Mice Body.Can be found out by scattergram, p-HA-micelles/DiR has obvious fluorescence distribution in mouse brain, and mPEG-micelles/DiR does not have fluorescence distribution in brain, prompting p-HA-micelles/DiR can be mediated and be penetrated blood brain barrier and enter brain (as shown in Figure 4) by P-hydroxybenzoic acid.
2) the mice in vitro tissue of P-hydroxybenzoic acid--polymer micelle/DiR distributes
Prepare respectively p-HA-micelles/DiR
,mPEG-micelles/DiR polymer micelle, wherein the consumption of DiR is 15.6 μ g, mouse tail vein injection 100 μ L/ only, respectively at after administration 2,4,8 and 12 hours with 10% chloral hydrate anesthesia mice, cardiac perfusion normal saline 100ml/ only, cores, the internal organs such as lung, liver,spleen,kidney and brain are placed in and in living animal imaging system, observe and carry out semiquantitative determination.Each internal organs fluorescence distribution result shows that p-HA-micelles/DiR has obvious fluorescence distribution in mouse brain, and prompting p-HA-micelles can be mediated and penetrated BBB and distribute (as shown in Figure 5) in brain by P-hydroxybenzoic acid.
embodiment 4
P-hydroxybenzoic acid-polymer micelle/DTX(p-HA-micelles/DTX) antigen position cerebral glioma pharmacodynamic evaluation
The take the logarithm U87 MG cell of trophophase, peptic cell counting, with appropriate PBS buffer suspension, every nude inoculation 8 × 10
5individual cell (being scattered in 5 μ L PBS buffer), experiment before by nude mice with after 7% chloral hydrate anesthesia, fix with brain solid positioner, suspension cell is inoculated in to striatum position;
By original position cerebral tumor model nude mice be divided at random 5 groups (
n=9), respectively at the 6th, 9,12,15 days injections 250 μ l normal saline (physiological saline), mPEG-micelles/DTX, p-HA-micelles/DTX and Texotere
?, the injected dose of single DTX is 8 mg/kg, the life span of record cast nude mice, and result shows the median survival time (as shown in Figure 6) that p-HA-micelles/ DTX can significant prolongation original position Brain Glioma Model Mus.
Claims (9)
1. the brain target polymer micelle delivery system of a P-hydroxybenzoic acid mediation, comprise targeted molecular, carrier and medicine, it is characterized in that, be jointly built into by thering is brain target function material (I), amphipathic nature polyalcohol material (II) and medicine the polymer micelle delivery system that wraps medicine carrying thing; Brain target function material (I) accounts for the 2%-20% of amphipathic nature polyalcohol material (II); Wherein: brain target function material (I) is made up of the mPEG2000-DSPE of modifying through P-hydroxybenzoic acid; Amphipathic nature polyalcohol material (II) is made up of the mPEG2000-DSPE of methoxyl group end-blocking.
2. by the brain target polymer micelle delivery system of P-hydroxybenzoic acid mediation claimed in claim 1, it is characterized in that, described P-hydroxybenzoic acid is brain targeted molecular.
3. by the brain target polymer micelle delivery system of P-hydroxybenzoic acid mediation claimed in claim 1, it is characterized in that, molecular weight polyethylene glycol in brain target function material is between 2000-5000, and the molecular weight polyethylene glycol in amphipathic nature polyalcohol material is between 1000-5000.
4. by the brain target polymer micelle delivery system of P-hydroxybenzoic acid mediation claimed in claim 1, it is characterized in that, PHOSPHATIDYL ETHANOLAMINE in brain target function material and amphipathic nature polyalcohol material is satisfied fatty acid PHOSPHATIDYL ETHANOLAMINE or unsaturated fatty acid PHOSPHATIDYL ETHANOLAMINE.
5. by the brain target polymer micelle delivery system of P-hydroxybenzoic acid mediation claimed in claim 1, it is characterized in that, in described delivery system, its medicine carrying mode is parcel or Electrostatic Absorption.
6. by the brain target polymer micelle delivery system of P-hydroxybenzoic acid mediation claimed in claim 1, it is characterized in that, the particle diameter of described polymer micelle delivery system is 7-30nm.
7. by the brain target polymer micelle delivery system of P-hydroxybenzoic acid mediation claimed in claim 1, it is characterized in that, the medicine that described delivery system is sent is one or more in diagnostic medicine or medicine;
Wherein, diagnostic medicine is selected from: nuclear medicine diagnostic medicine, MR diagnosis medicine or lipotropy antitumor drug.
8. by the brain target polymer micelle delivery system of P-hydroxybenzoic acid mediation claimed in claim 1, it is characterized in that, described lipotropy antitumor drug is selected from, one or both in paclitaxel, Docetaxel, amycin, camptothecine, hydroxy camptothecin, 9-nitrocamptothecin or levodopa.
9. the preparation method of the brain target polymer micelle delivery system of the P-hydroxybenzoic acid of claim 1 mediation, it is characterized in that, adopt thin film aquation legal system for polymer micelle delivery system, wherein: respectively brain target function material (I), amphipathic nature polyalcohol material (II) and medicine are dissolved in to organic solvent altogether, after film forming, add aquation medium aquation to form polymer micelle; Its midbrain target function material (I) is mol ratio 5%, 10%, 15% or 20% with the ratio of amphipathic nature polyalcohol material (II); Described medicine is one or more in diagnostic medicine or medicine; Wherein, diagnostic medicine is selected from: nuclear medicine diagnostic medicine, MR diagnosis medicine or lipotropy antitumor drug.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104382851A (en) * | 2014-10-17 | 2015-03-04 | 南开大学 | Method for preparing intelligent target medicine carrying composite micelles |
| CN107714646A (en) * | 2017-10-26 | 2018-02-23 | 苏州大学 | Amphipathic nature polyalcohol micella of tumor extracellular matrix and preparation method thereof can be penetrated |
| CN110628011A (en) * | 2018-06-21 | 2019-12-31 | 中国医学科学院药物研究所 | Phospholipid-polyethylene glycol-borneol polymer, and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101721350A (en) * | 2008-10-10 | 2010-06-09 | 复旦大学 | Folate-mediated targeted polymeric micelle |
| CN102160894A (en) * | 2010-02-22 | 2011-08-24 | 复旦大学 | Benzamide analog mediated brain-targeting delivery system |
-
2013
- 2013-02-07 CN CN201310050084.5A patent/CN103977434B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101721350A (en) * | 2008-10-10 | 2010-06-09 | 复旦大学 | Folate-mediated targeted polymeric micelle |
| CN102160894A (en) * | 2010-02-22 | 2011-08-24 | 复旦大学 | Benzamide analog mediated brain-targeting delivery system |
Non-Patent Citations (2)
| Title |
|---|
| 张智新: "基于对羟基苯甲酸和肉豆蔻介导的脑靶向聚合物胶束递送系统研究", 《中国优秀硕士学位论文全文数据库（电子期刊）》 * |
| 王朝辉 等: "脑靶向纳米载体系统的研究进展", 《中国生化药物杂志》 * |
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| CN104382851A (en) * | 2014-10-17 | 2015-03-04 | 南开大学 | Method for preparing intelligent target medicine carrying composite micelles |
| CN104382851B (en) * | 2014-10-17 | 2018-12-28 | 南开大学 | A kind of preparation method of intelligence target drug-carrying composite micelle |
| CN107714646A (en) * | 2017-10-26 | 2018-02-23 | 苏州大学 | Amphipathic nature polyalcohol micella of tumor extracellular matrix and preparation method thereof can be penetrated |
| CN110628011A (en) * | 2018-06-21 | 2019-12-31 | 中国医学科学院药物研究所 | Phospholipid-polyethylene glycol-borneol polymer, and preparation method and application thereof |
| CN110628011B (en) * | 2018-06-21 | 2021-12-14 | 中国医学科学院药物研究所 | Phospholipid-polyethylene glycol-borneol polymer, and preparation method and application thereof |
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