CN102389573A - Medicine for treating malignant glioma and preparation method thereof - Google Patents
Medicine for treating malignant glioma and preparation method thereof Download PDFInfo
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- 208000029824 high grade glioma Diseases 0.000 title abstract 4
- 201000011614 malignant glioma Diseases 0.000 title abstract 4
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Abstract
The invention discloses a medicament for treating malignant glioma, which takes RGD polypeptide as an effective component and a polyaspartic acid ion compound as a carrier to treat malignant glioma in a targeted manner. The nanometer preparation can smoothly penetrate blood brain barrier and hematoma barrier, target RGD to gather in brain glioma, and specifically induce apoptosis of glioma cells. The preparation method provided by the invention is simple and feasible, the raw materials are easy to obtain, the repeatability of the preparation process is good, the prepared nano-micelle is good in stability, can stably exist under different pH values and salt concentrations, does not damage the structure and the function of RGD polypeptide, can well guarantee the biological activity of RGD, and can effectively treat malignant glioma.
Description
Technical field
The invention belongs to the medical preparation field, more particularly, relate to a kind of nanometer formulation of treating glioblastoma.
Background technology
Cerebral glioma is the modal malignant tumor of people central nervous system; Recently research shows; Glioma is the blood vessel dependent tumors; In the propagation and invasive procedure of glioma cell and vascular endothelial cell, the specific high expressed integrin alpha v beta 3 of glioma cell and vascular endothelial cell (INT α v β 3), and expression intensity obviously strengthens along with increasing of cerebral glioma pathology rank.In the neovascular tissue of tumor inducing, 3 pairs of tumor vascular generations of INT α v β of expressed in abundance play an important role, and participate in the migration and the invasion and attack of most of glioblastoma.Though tissue has the expression of INT α v β 3 outside normal cerebral tissue and cranium, expression is very low.Rgd peptide is as integrating recognition site plain and its ligand interaction; Sticking each other between cell and cell, cell and extracellular matrix (ECM); And play critical effect in the transduction of the two-way signaling between cell and the ECM, regulated and control sticking, breed, break up, shift and transferring and die of tumor cell.The courier who integrates interaction that plain recognition site suppresses endotheliocyte and ECM, blocking-up blood vessel epithelial cell proliferation through competition transmits, stops cell proliferation, suppresses tumor neovasculature formation, the migration and the invasive growth of prevention tumor cell.In addition; The RGD peptide of high concentration can through and cell in a potential RGD binding sequence self containing of procaspase-3 be the interaction between the Asp-Asp-Met (DDM); Cause procaspase-3 conformational changes; Thereby activate caspase-3, directly the accent of inducing tumor cell is died.
But because the long peptide of most of linear RGD is difficult to cross over blood brain barrier, local dose is low excessively, and RGD small peptide and becate peptide circulating half-life are short, are easy to remove, and are difficult to arrive the effect of the treatment cerebral tumor.Therefore, the RGD peptide only can be used to prepare cerebral tumor developing agent at present, and aspect the treatment of the cerebral tumor, does not report as yet both at home and abroad.
Drug-carried nanometer is the bonded product of nanotechnology and modern medicine and pharmacology, is the novel form of a kind of medicine controlled releasing and slow release.It comprises nanoparticle (nanopart icales) and Nano capsule (nanocap su les); Be the solid shape colloidal particle of diameter between 10～500nm; Active part (medicine, bioactive materials etc.) is positioned at particle inside through dissolving, package action, perhaps is positioned at particle surface through absorption, adhewsive action.Since Birrenbach reported first drug-carried nanometer in 1976, nanoparticle is applied in the medical domain as medicament slow release and targeting vector widely.Wherein, polyion micelle has been avoided the use of organic solvent owing to can in aqueous solution, carry out medicine carrying, receives extensive concern.The stability of polyion micelle receives the influence of various factorss such as chemical composition of micelle own and environment of living in thereof.Crosslinked through to polyion micelle can improve the stability of effective raising polyion micelle.
Summary of the invention
First purpose of the present invention is to provide a kind of medicine of treating glioblastoma.
Second purpose of the present invention is to provide a kind of method for preparing of treating the medicine of glioblastoma.
The 3rd purpose of the present invention is to provide a kind of nervous system pharmaceutical carrier.
The 4th purpose of the present invention is to provide a kind of application of poly-aspartate ion complex micelle in the medicine of preparation treatment glioblastoma of rgd peptide modification.
The 5th purpose of the present invention is to provide the application as carrier in the medicine of preparation treatment glioblastoma of a kind of poly-aspartate ion complex.
For realizing first purpose of the present invention, disclose following technical scheme: a kind of medicine of treating glioblastoma, said medicine are the poly-aspartate ion complex micelle that rgd peptide is modified.
Said poly-aspartate ion complex is with the end capped Polyethylene Glycol polycaprolactone of maleimide base group copolymer; Said copolymer forms nanoparticle according to mol ratio 1:1 through the electrostatic interaction self assembly by poly-aspartate derivant PEG-g-PDEA and PEG-g-PAsp; Wherein electronegative PAsp and electropositive PDEA electrostatic attraction form hydrophobic core, and skin is made up of biocompatible hydrophilic polymer PEG.
For realizing second purpose of the present invention, disclose following technical scheme: the method for preparing of the medicine of treatment glioblastoma may further comprise the steps:
(a) the preparation end group is the Polyethylene Glycol Mal-PEG-NH of dimaleoyl imino and amido
2
(b) preparation Polyethylene Glycol grafting polysuccinimide PEG-g-PSI;
(c) preparation poly-aspartate derivant PEG-g-PDEA;
(d) preparation PEG-g-PAsp;
(e) preparation poly-aspartate base polyion micelle PIC and crosslinked;
(f) preparation RGD-PIC.
Wherein, (a) end group is the (Mal-PEG-NH of the Polyethylene Glycol of dimaleoyl imino and amido
2) preparation: because the terminal hydroxy group of Mal-PEG-OH has certain activity, utilize
N, N-two succinimidyl carbonates (DSC) activation Mal-PEG-OH is intermediate Mal-PEG-SC, utilizes excessive ethylenediamine to replace SC again and directly obtains Mal-PEG-NH
2, reaction equation is as shown in Figure 1.
(b) preparation of Polyethylene Glycol grafting polysuccinimide (PEG-g-PSI): utilize Mal-PEG-NH
2With the polysuccinimide ring-opening reaction, two kinds have been prepared respectively with the poly-aspartate derivant PEG-that has maleimide base group of positive and negative electric charge
g-PDEA and PEG-
g-PAsp, the course of reaction sketch map is as shown in Figure 2.
(c) preparation of poly-aspartate derivant PEG-g-PDEA: PEG-g-PSI is scattered in the low amounts of water, magnetic agitation 10 min; Slow dropping sodium aqueous solution after dropwising, reacts 6 h at normal temperatures under the ice bath to PEG-g-PSI solution; After reaction finishes, add 0.1 M HCl regulator solution pH value to neutral; Dialysis, lyophilization obtains white solid powder PEG-
g-PAsp; The group of infrared test assay products changes situation.
(d) preparation of PEG-g-PAsp: 1 g PEG-g-PSI is dissolved in 10 mL DMF; Solution slowly is added drop-wise in the 5 mL ethylenediamines, reacts 8 h at normal temperatures; After reaction finishes,, obtain the light yellow solid powder with the absolute ether deposition; Absolute ethanol washing is removed excessive ethylenediamine for several times again; End product is 40
oC vacuum drying 24 h obtain light yellow solid powder PEG-
g-PDEA; The group of infrared test assay products changes situation.
(e) preparation of poly-aspartate base polyion micelle and crosslinked:
(1) to be dissolved in pH value respectively be that the mol ratio of PEG-g-PDEA and PEG-g-PAsp and phosphate buffer is 1:2 in 7.4 the phosphate buffer for PEG-g-PDEA and PEG-g-PAsp;
(2) under the magnetic agitation PEG-g-PAsp solution slowly is added drop-wise in the PEG-g-PDEA solution, the mol ratio of PEG-g-PAsp solution and PEG-g-PDEA solution is 1:1;
(3) continue to stir half an hour, with 0.45 μ m water membrane filtration;
(4) filtrating slowly adds 80-120 μ l 2.5% glutaraldehyde water solution under stirring condition;
(5) stirring at normal temperature is reacted 4 h, with 0.45 μ m water membrane filtration, obtains end group maleimide amination poly-aspartate base polyion micelle Mal-PIC.
(f) preparation of RGD-PIC: the Mal-PIC that step (e) obtains and C (RGDfC) five rings peptide are according to maleimide base group: sulfydryl mixes for mol ratio 3:1, and the lucifuge magnetic agitation is reacted and spent the night; Reacted solution is crossed 1.5 * 20 cm, and Sepharose CL-4B post is removed unconjugated C (RGDfC); Using 0.01 mol/L pH value is 7.4 PBS buffer solution elution, collects the RGD-PIC suspension of white.
Wherein, step (a) and (b), (c) and (d) be the conventional method preparation.
For realizing the 3rd purpose of the present invention; Following technical scheme is disclosed: a kind of nervous system pharmaceutical carrier; Said carrier is with the end capped Polyethylene Glycol polycaprolactone of maleimide base group copolymer; Said copolymer forms nanoparticle according to mol ratio 1:1 through the electrostatic interaction self assembly by poly-aspartate derivant PEG-g-PDEA and PEG-g-PAsp, and wherein electronegative PAsp and electropositive PDEA electrostatic attraction form hydrophobic core, and skin is made up of biocompatible hydrophilic polymer PEG.
For realizing the 4th purpose of the present invention; Following technical scheme is disclosed: the application of the poly-aspartate ion complex micelle that rgd peptide is modified in the medicine of preparation treatment glioblastoma; Said poly-aspartate ion complex is with the end capped Polyethylene Glycol polycaprolactone of maleimide base group copolymer; Said copolymer forms nanoparticle according to mol ratio 1:1 through the electrostatic interaction self assembly by poly-aspartate derivant PEG-g-PDEA and PEG-g-PAsp; Wherein electronegative PAsp and electropositive PDEA electrostatic attraction form hydrophobic core, and skin is made up of biocompatible hydrophilic polymer PEG.
For realizing the 5th purpose of the present invention; Following technical scheme is disclosed: the poly-aspartate ion complex in the medicine of preparation treatment glioblastoma as the application of carrier; Said poly-aspartate ion complex is with the end capped Polyethylene Glycol polycaprolactone of maleimide base group copolymer; Said copolymer forms nanoparticle according to mol ratio 1:1 through the electrostatic interaction self assembly by poly-aspartate derivant PEG-g-PDEA and PEG-g-PAsp; Wherein electronegative PAsp and electropositive PDEA electrostatic attraction form hydrophobic core, and skin is made up of biocompatible hydrophilic polymer PEG.
The invention has the advantages that: medicine provided by the invention is active ingredient with the rgd peptide, is carrier with the high molecular polymer, the targeted therapy glioblastoma.This nanometer formulation can see through blood brain barrier and hematoma tumor barrier smoothly; The RGD targeting is gathered in glioma in the brain, and the specific apoptosis that causes glioma cell is not owing to contain the other drug composition; This nanometer formulation does not have other toxic and side effects, and for the not influence of tumor Zhou Zhengchang tissue.Method for preparing provided by the invention is simple; Raw material is easy to get, preparation process good reproducibility, and the nano-micelle that makes has good stability; Can be under different PH and salinity stable existence; And do not destroy the 26S Proteasome Structure and Function of rgd peptide, can well ensure the BA of RGD, effectively treat glioblastoma.
Description of drawings
Fig. 1 is the synthetic sketch map of Mal-PEG-NH2.
Fig. 2 is the synthetic sketch map of poly-aspartate derivant PEG-g-PDEA and PEG-g-PAsp.
Fig. 3 is c (RGDfC) five rings peptide structural formula.
Fig. 4 is the relative light intensity value of DLS test RGD-PIC under the different PH environment.
Fig. 5 is the influence of RGD-PIC to hippocampus of rats cells survival rate.
Fig. 6 is the influence of the RGD-PIC of variable concentrations for people's glioma cell survival rate.
Fig. 7 is that mtt assay detected each experimental group cells survival rate after the RGD-PIC of use various dose handled rat cortical neuron and people's glioma cell U87 respectively.
The specific embodiment
Below in conjunction with accompanying drawing the present invention is elaborated, the effect of embodiment only is to explain and non-limiting the present invention.
The preparation 1 of the poly-aspartate ion complex micelle RGD-PIC that embodiment 1, rgd peptide are modified
In the 100mL round-bottomed flask, add 10g (5mmol) Mal-PEG
nThe DMF dissolving of heavily steaming with 40mL obtains colourless transparent solution; Add rapidly 7.68g (12mmol) triethylamine under the magnetic agitation, obtain the Polyethylene Glycol succinimdyl carbonate (PEGn-SC) that white solid state has the Mal group after ice ether and 60 ℃ remove the water-toluene purification.In the 50ml round-bottomed flask, add 10mL dewater dichloromethane, 3.06mL (46mmol) ethylenediamine; The 10mL dissolved 5g of dichloromethane (2.3mmol) PEGn-SC that dewaters; Normal-temperature reaction 12h; Reaction finishes after-filtration, deposition recrystallization, 40 ℃ of dry 24h, and obtaining the white solid state end group at last is dimaleoyl imino and aminated Polyethylene Glycol (MAL-PEG
n-NH2) 4g, productive rate is 80%, (n=1000 ~ 50000).
Add 10g L-aspartic acid in the 150mL three-necked bottle, 0.89g phosphoric acid (85%), 9.5g sulfolane and 22g 1,3,5-trimethylbenzene.Elevated temperature under nitrogen protection, 180 ℃ of backflow 4.5h.Reaction finishes after-filtration, separates, and washing with acetone is removed the sulfolane and 1 on surface for 3 times; 3,5-trimethylbenzene, an amount of DMF dissolving; Remove by filter unreacted L-aspartic acid, deionized water deposition, sucking filtration; Centrifuge washing, 60 ℃ of vacuum dryings of product 24 hours obtain white solid state polysuccinimide (PSI).
10mLDMF dissolves a certain amount of MAL-PEG respectively
n-NH2, PSI, 60 ℃ of magnetic agitation 8h, ice ether sedimentation, THF washing; Remove unreacted MAL-PEGn-NH2, end product is 40 ℃ of dry 24h under vacuum, obtain white solid state end group maleimide polyethylene glycol grafting polysuccinimide (MAL-PEG-g-PSI); Get 1g and be dispersed in 10mL water, magnetic agitation 10mins, slow Dropwise 5 g20% sodium hydrate aqueous solution under the ice bath; After dropwising, react 6h at normal temperatures, after this add 0.1MHCl regulator solution pH value to neutral; Behind the above-mentioned solution 48h of MWCO 3500 dialysis, lyophilization obtains end group maleimide polyethylene glycol grafting poly-aspartate (MAL-PEG-g-PAsp).Other gets 1g and is scattered among the 10mLDMF; Add the 5mL ethylenediamine,, behind the reaction 8h; The absolute ether deposition; Absolute ethanol washing is removed the excessive ethylenediamine of unreacted, and end product obtains end group maleimide polyethylene glycol grafting polyethyene diamine base agedoite (MAL-PEG-g-PDEA) at 40 ℃ of dry 24h.Respectively take by weighing 5mMPEG-g-PDEA and PEG-g-PAsp; Be dissolved in phosphate buffer (pH=7.4) magnetic agitation of 10mL10mM; Filter; Slowly add a certain amount of glutaraldehyde (2.5% aqueous solution), continue stirring at normal temperature 4h, membrane filtration gets end group maleimide amination poly-aspartate base polyion micelle (Mal PIC).
Mal-PIC mixes according to maleimide base group/sulfydryl=3/1 with C (RGDfC), and the reaction of lucifuge magnetic agitation is spent the night; Reacted solution is crossed 1.5 * 20 cm, and Sepharose CL-4B post is removed unconjugated C (RGDfC); With 0.01 mol/L PBS buffer (pH 7.4) eluting, collect the RGD-PIC suspension of white.
The preparation 2 of the poly-aspartate ion complex micelle RGD-PIC that embodiment 2, rgd peptide are modified
In the 100mL round-bottomed flask, add 10g (5mmol) Mal-PEG
n, the DMF dissolving of heavily steaming with 40mL obtains colourless transparent solution, adds 8.35mL (24mmol) triethylamine under the magnetic agitation rapidly, after reaction finishes, obtains the PEG that white solid state has the Mal group after removing the water-toluene purification through ice ether and 60 ℃ repeatedly
n-SC.In the 50ml round-bottomed flask, add 10mL dewater dichloromethane, 3.06mL (46mmol) ethylenediamine; The 10mL dissolved 5g of dichloromethane (2.3mmol) PEGn-SC that dewaters; Normal-temperature reaction 12h; Reaction finishes after-filtration, deposition recrystallization, 40 ℃ of dry 24h, and obtaining the white solid state end group at last is dimaleoyl imino and aminated Polyethylene Glycol (MAL-PEG
n-NH2) 4g, productive rate is 80%, (n=1000 ~ 50000).
Add 10g L-aspartic acid in the 150mL three-necked bottle, 0.89g phosphoric acid (85%), 9.5g sulfolane and 22g 1,3,5-trimethylbenzene.Elevated temperature under nitrogen protection, 180 ℃ of backflow 4.5h.Reaction finishes after-filtration, separates, and washing with acetone is removed the sulfolane and 1 on surface for 3 times; 3,5-trimethylbenzene, an amount of DMF dissolving; Remove by filter unreacted L-aspartic acid, deionized water deposition, sucking filtration; Centrifuge washing, 60 ℃ of vacuum dryings of product 24 hours obtain white solid state polysuccinimide (PSI).
10mLDMF dissolves a certain amount of MAL-PEG respectively
n-NH2, PSI, 60 ℃ of magnetic agitation 8h, ice ether sedimentation, THF washing; Remove unreacted MAL-PEGn-NH2, end product is 40 ℃ of dry 24h under vacuum, obtain white solid state end group maleimide polyethylene glycol grafting polysuccinimide (MAL-PEG-g-PSI); Get 1g and be dispersed in 10mL water, magnetic agitation 10mins, slow Dropwise 5 g20% sodium hydrate aqueous solution under the ice bath; After dropwising, react 6h at normal temperatures, after this add 0.1MHCl regulator solution pH value to neutral; Behind the above-mentioned solution 48h of MWCO 3500 dialysis, lyophilization obtains end group maleimide polyethylene glycol grafting poly-aspartate (MAL-PEG-g-PAsp).Other gets 1g and is scattered among the 10mLDMF; Add the 5mL ethylenediamine,, behind the reaction 8h; The absolute ether deposition; Absolute ethanol washing is removed the excessive ethylenediamine of unreacted, and end product obtains end group maleimide polyethylene glycol grafting polyethyene diamine base agedoite (MAL-PEG-g-PDEA) at 40 ℃ of dry 24h.Respectively take by weighing 5mMPEG-g-PDEA and PEG-g-PAsp; Be dissolved in phosphate buffer (pH=7.4) magnetic agitation of 10mL10mM; Filter; Slowly add a certain amount of glutaraldehyde (2.5% aqueous solution), continue stirring at normal temperature 4h, membrane filtration gets end group maleimide amination poly-aspartate base polyion micelle (Mal PIC).
Mal-PIC mixes according to maleimide base group/sulfydryl=3/1 with C (RGDfC), and the reaction of lucifuge magnetic agitation is spent the night; Reacted solution is crossed 1.5 * 20 cm, and Sepharose CL-4B post is removed unconjugated C (RGDfC); With 0.01 mol/L PBS buffer (pH 7.4) eluting, collect the RGD-PIC suspension of white.
The stability test of embodiment 3, RGD-PIC
As shown in Figure 4, we have studied the stable case of the RGD-PIC solution under the different PH environment, and selected pH value scope is 6.5-12; In the PB buffer, prepare polyion micelle solution; Regulate different pH, DLS tests with dynamic light scattering, and the light intensity of solution under the different pH is studied; Choosing the maximum optical strength is standard point, with relative light intensity object of study the most.Can see as pH 8.5 the time, the relative light intensity value of the RGD-PIC that DSL records weakens, and is 66.73%, and reduces along with the rising of pH, explains that polyion micelle is unstable.When pH value reaches neutrality (7-8), the relative light intensity value of RGD-PIC remains on more than 90%.The stability that shows RGD-PIC can receive the influence of body fluid pH value hardly under the body fluid environment.
The biocompatibility test of embodiment 4, RGD-PIC
The test cell line results suggest, the present invention all has excellent biological compatibility for rat nerves unit, rat star spongiocyte, rat microglia, people's brain capillary endothelial cell.Detection for rat nerves unit survival rate is as shown in Figure 5.
The rat cortical neuron is former is commissioned to train to support and is divided into 12 groups at random after 7 days, gives 0umol/ml respectively, 50 umol/ml; 150 umol/ml; The PIC of 300 umol/ml, c (RGDfC) and RGD-PIC handled 4 hours, and mtt assay detects each experimental group hippocampus of rats cells survival rate.Wherein, the negative matched group of PIC, the positive matched group of c (RGDfC).The result shows that each organizes rat nerves unit cells survival rate there was no significant difference.
The medicinal effects test of embodiment 5, RGD-PIC
The present invention has lethal effect for glioma cell, and the result is as shown in Figure 6.Cultivating people's glioma cell U87.Be divided into 12 groups at random, give 0umol/ml respectively, 50 umol/ml, 150 umol/ml, the PIC of 300 umol/ml, c (RGDfC) and RGD-PIC handled 4 hours, the negative matched group of PIC, the positive matched group of c (RGDfC).Mtt assay detects each experimental group cells survival rate.The result shows RGD-PIC processed group glioma cell survival rate, and there were significant differences than the PIC group, and have the agent effect relationship.Heavy dose of group smaller dose group better effects if.
Can know that by Fig. 7 after the RGD-PIC of use various dose handled rat cortical neuron and people's glioma cell U87 respectively, mtt assay detected each experimental group cells survival rate.The result shows, rat cortical neuron cell and U87 cells survival rate obvious difference that the RGD-PIC of same dose handles.RGD-PIC has tangible lethal effect for glioma cell, and does not have what influence for the survival rate of normal neurons cell.
The above only is a preferred implementation of the present invention; Should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the principle of the invention; Can also make some improvement and retouching, these improvement and retouching also should be regarded as protection scope of the present invention.
Claims (8)
1. a medicine of treating glioblastoma is characterized in that, said medicine is the poly-aspartate ion complex micelle that rgd peptide is modified.
2. the medicine of treatment glioblastoma according to claim 1; It is characterized in that; Said poly-aspartate ion complex is with the end capped Polyethylene Glycol polycaprolactone of maleimide base group copolymer; Said copolymer forms nanoparticle according to mol ratio 1:1 through the electrostatic interaction self assembly by poly-aspartate derivant PEG-g-PDEA and PEG-g-PAsp; Wherein electronegative PAsp and electropositive PDEA electrostatic attraction form hydrophobic core, and skin is made up of biocompatible hydrophilic polymer PEG.
3. the method for preparing of the medicine of the described treatment glioblastoma of claim 1 is characterized in that, may further comprise the steps:
(a) the preparation end group is the Polyethylene Glycol Mal-PEG-NH of dimaleoyl imino and amido
2
(b) preparation Polyethylene Glycol grafting polysuccinimide PEG-g-PSI;
(c) preparation poly-aspartate derivant PEG-g-PDEA;
(d) preparation PEG-g-PAsp;
(e) preparation poly-aspartate base polyion micelle PIC and crosslinked;
(f) preparation RGD-PIC.
4. the method for preparing of the medicine of treatment glioblastoma according to claim 3 is characterized in that, the preparation of step (e) poly-aspartate base polyion micelle may further comprise the steps:
(1) to be dissolved in pH value respectively be that the mol ratio of PEG-g-PDEA and PEG-g-PAsp and phosphate buffer is 1:2 in 7.4 the phosphate buffer for PEG-g-PDEA and PEG-g-PAsp;
(2) under the magnetic agitation PEG-g-PAsp solution slowly is added drop-wise in the PEG-g-PDEA solution, the mol ratio of PEG-g-PAsp solution and PEG-g-PDEA solution is 1:1;
(3) continue to stir half an hour, with 0.45 μ m water membrane filtration;
(4) filtrating slowly adds 80-120 μ l 2.5% glutaraldehyde water solution under stirring condition;
(5) stirring at normal temperature is reacted 4 h, with 0.45 μ m water membrane filtration, obtains end group maleimide amination poly-aspartate base polyion micelle Mal-PIC.
5. the method for preparing of the medicine of treatment glioblastoma according to claim 3 is characterized in that, the preparation of step (f) RGD-PIC may further comprise the steps:
The Mal-PIC that step (e) obtains and C (RGDfC) five rings peptide are according to maleimide base group: sulfydryl mixes for mol ratio 3:1, and the lucifuge magnetic agitation is reacted and spent the night; Reacted solution is crossed 1.5 * 20 cm, and Sepharose CL-4B post is removed unconjugated C (RGDfC); Using 0.01 mol/L pH value is 7.4 PBS buffer solution elution, collects the RGD-PIC suspension of white.
6. nervous system pharmaceutical carrier; It is characterized in that; Said carrier is with the end capped Polyethylene Glycol polycaprolactone of maleimide base group copolymer; Said copolymer forms nanoparticle according to mol ratio 1:1 through the electrostatic interaction self assembly by poly-aspartate derivant PEG-g-PDEA and PEG-g-PAsp, and wherein electronegative PAsp and electropositive PDEA electrostatic attraction form hydrophobic core, and skin is made up of biocompatible hydrophilic polymer PEG.
7.RGD the application of peptide modified poly-aspartate ion complex micelle in the medicine of preparation treatment glioblastoma; It is characterized in that; Said poly-aspartate ion complex is with the end capped Polyethylene Glycol polycaprolactone of maleimide base group copolymer; Said copolymer forms nanoparticle according to mol ratio 1:1 through the electrostatic interaction self assembly by poly-aspartate derivant PEG-g-PDEA and PEG-g-PAsp; Wherein electronegative PAsp and electropositive PDEA electrostatic attraction form hydrophobic core, and skin is made up of biocompatible hydrophilic polymer PEG.
The poly-aspartate ion complex in the medicine of preparation treatment glioblastoma as the application of carrier; It is characterized in that; Said poly-aspartate ion complex is with the end capped Polyethylene Glycol polycaprolactone of maleimide base group copolymer; Said copolymer forms nanoparticle according to mol ratio 1:1 through the electrostatic interaction self assembly by poly-aspartate derivant PEG-g-PDEA and PEG-g-PAsp; Wherein electronegative PAsp and electropositive PDEA electrostatic attraction form hydrophobic core, and skin is made up of biocompatible hydrophilic polymer PEG.
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| CN110028674A (en) * | 2018-01-12 | 2019-07-19 | 中国科学院苏州纳米技术与纳米仿生研究所 | A kind of amphipathy macromolecule material system, preparation method and application |
| CN111494644A (en) * | 2020-06-19 | 2020-08-07 | 首都医科大学 | Nanometer carrier containing RGD sequence peptide, preparation method thereof, drug-loading system, preparation method and application thereof |
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| 《中国博士学位论文全文数据库》 20091115 刘晓英 "脑靶向载药纳米胶束治疗胶质瘤和耐药性癫痫的实验研究" 摘要，第12－18页 1-8 , * |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110028674A (en) * | 2018-01-12 | 2019-07-19 | 中国科学院苏州纳米技术与纳米仿生研究所 | A kind of amphipathy macromolecule material system, preparation method and application |
| CN111494644A (en) * | 2020-06-19 | 2020-08-07 | 首都医科大学 | Nanometer carrier containing RGD sequence peptide, preparation method thereof, drug-loading system, preparation method and application thereof |
| CN111494644B (en) * | 2020-06-19 | 2022-08-19 | 首都医科大学 | Nano-carrier containing RGD sequence peptide, preparation method thereof, drug-loading system, preparation method and application thereof |
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