CN106880846A - A kind of cancer target multifunctional nano delivery system and preparation method and purposes - Google Patents
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Abstract
The invention discloses a kind of cancer target multifunctional nano delivery system and preparation method and purposes, its system includes the multifunctional nano grain of tumour penetrating peptide RGERPPR modifications and antineoplastic, and the multifunctional nano grain is T1 T2 bimodulus magnetic resonance contrast agents；The antineoplastic is doxorubicin or epirubicin；Percentage by weight is 10% 35% to its antineoplastic in systems.The delivery system has following characteristics：Particle diameter is spherical in shape in 60 200nm；By T1 contrast agent Gd DTPA and T2 contrast agent Fe3O4Utilize simultaneously, there is provided the diagnostic message of pin-point accuracy；The water solubility and biocompatibility of medicine are improve, the curative effect and bioavilability of medicine is improve；Can be used to be injected intravenously, by RGERPPR mediations and tumour EPR effects targeted to tumor tissues, the system has obvious inhibiting effect to glioma tumour growth；The delivery system can be used for the diagnosis and treatment integration of diagnosing tumor and oncotherapy.
Description
Technical field
The invention belongs to field of pharmaceutical preparations, it is related to a kind of peptide modified targeted nano delivery system, specifically swells
The active targeting multifunctional nano delivery system and preparation method and purposes of knurl penetrating peptide RGERPPR modifications.
Background technology
Cancer is the major public health problem that most countries face in the world, is the world's second largest cause of death.Mesh
Before, chemotherapy is still one of most efficient method for the treatment of cancer in clinic.However, because conventional chemotherapeutics is in body
It is interior cannot be by real-time spike, it cannot learn in the distribution situation of tumor tissues, thus cannot be medicine Clinical efficacy carry
For feedback in time.This is also that chemotherapy of tumors cannot realize the main cause of individualized treatment, is considered current by many scientists
The subject matter that classic chemotherapy clinically faces, is badly in need of a kind of effective technology and solves.
The diagnosis and treatment integration function of cancer diagnosis and treatment of cancer, has become the new trend for the treatment of of cancer.With
The fast development of nano science and nanometer technology, the foundation of nanosecond medical science and develop into the early diagnosis of the principal diseases such as cancer
A new multifunctional platform is provided with precisely treatment.Novel nano probe can integrate the function of diagnosis and treatment, can be with
The imaging and treatment of tumour are realized simultaneously.
Magnetic resonance imaging (MRI) is a kind of non-invasive diagnostic techniques, clinical at present with spatial resolution higher
On be widely used in tumor imaging.MRI contrast agent is commonly used to strengthen the contrast between normal structure and pathological tissues, so that
Improve the accuracy of diagnosis.By development for many years, contrast agent is roughly divided into T1 contrast agent and T2 contrast agent, current T1 radiographies
Agent mainly provides brighter image, and T2 contrast agent is typically to produce darker image.Although the development of T1 and T2 contrast agent
Have been achieved for successfully, but the contrast agent of this single-mode respectively has advantage and disadvantage, increasingly faces new challenges.Therefore, it is right
The exploitation of the double mode NMR contrast agents of T1-T2 is very attractive because two different imaging patterns (T1 and
T2) can utilize simultaneously, there is provided the diagnostic message of pin-point accuracy.
Adriamycin is one of conventional antineoplastic, and antitumor spectra is relatively broad, belongs to cell cycle nonspecific agent (CCNSA),
Can all there is lethal effect to the tumour cell in each growth cycle.But it is secondary that its clinical efficacy is limited by serious cardio-toxicity grade
Effect, therefore therapeutic effect is not too preferable.Targeted drug delivery system, especially ligand modified tumor-targeting pass medicine
System, due to its accurate cancer target ability, enhances antitumor efficacy and receives the advantages of reduce whole body toxic and side effect
To increasing concern.Targeting ligand such as peptide, antibody and its fragment, play a crucial role in cancer target, its by with it is swollen
Specific expressed acceptor interaction in tumor tissue and mediate the cell endocytic of nanoscale medicine delivery system.NPR-1 is one kind in glue
There is the acceptor of expression high on matter blastoma and tumor vascular endothelial cell surface.RGERPPR polypeptides, the specificity of NPR-1 is matched somebody with somebody
Body, is a kind of tumour penetrating peptide, with the ability for penetrating tumor vessel wall and tumor tissues.The delivery system of RGERPPR modifications
Combined with tumor vascular endothelial cell surface and tumor cell surface NPR-1 receptor-specifics, penetrated tumor vessel wall and tumour
Cell, into tumour cell, plays the neoplasm growth effect of adriamycin, realizes the mesh to glioblastoma targeted therapy
's.And the tumor-selective of delivery system can be improved, pharmacokinetics behavior is improved, tolerance dose is increased, pair is reduced
Effect, and improve antitumous effect.
The content of the invention
Lack the problem of spike, weak curative effect in vivo it is an object of the invention to be directed to delivery system, there is provided a kind of tumour
The cancer target multifunctional nano delivery system of penetrating peptide RGERPPR modifications, Fe3O4/SiO2/mSiO2/DOX-(Gd-DTPA)-
PEG-RGE is used for the diagnosis and treatment integration of diagnosing tumor and oncotherapy.Delivery system Fe of the invention3O4/SiO2/mSiO2/
DOX- (Gd-DTPA)-PEG-RGE can be administered by being injected intravenously, and be targetted by RGERPPR mediations and tumour EPR effects
To tumor tissues, tumour cell is killed, in addition, it is by T1 contrast agent Gd-DTPA and T2 contrast agent Fe3O4Utilize simultaneously, Ke Yitong
Magnetic resonance imaging spike Nano medication is crossed, there is provided the diagnostic message of pin-point accuracy, so as to realize diagnosing tumor and oncotherapy
Diagnosis and treatment integration.
The object of the present invention is achieved like this：
A kind of cancer target multifunctional nano delivery system, Fe3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE,
Feature is that the multifunctional nano grain that the delivery system is modified by tumour penetrating peptide RGERPPR and antineoplastic are constituted, described
Multifunctional nano grain is T1-T2 bimodulus magnetic resonance contrast agents；The antineoplastic is doxorubicin or epirubicin；Its is antitumor
Percentage by weight is 10%-35% to medicine in systems.Its structure is as follows：
Wherein：Delivery system is nanoparticle, and in 60-200nm, particle diameter distribution PDI is 0.10-0.60 to particle diameter, and n is polymerization
Degree, n=9-227.
A kind of preparation method of above-mentioned nanoscale medicine delivery system, the method includes step in detail below：
Step 1：Prepare Fe3O4Nano-particle
1.39-5.56g iron oleates add 0.24-0.96mL octadecylenes after 30 DEG C of vacuum drying chambers stand 24 hours；It is mixed
Compound is with 3.3 DEG C of min of heating rate-1320 DEG C are heated to, reaction 60 minutes is then kept at this temperature.After reaction, it is cooled to
Room temperature, through 5-20mL n-hexanes and the washing of 20-80mL acetone, centrifugation, obtains Fe3O4Nano-particle；
Step 2：Prepare Fe3O4/SiO2Nano-particle
3.4-13.6mg Fe3O4Nano-particle and 4-16mL Igepal CO-520 are dissolved in 85-340mL hexamethylenes,
Excusing from death 30 minutes.Then, 0.65-2.6mL ammoniacal liquor is added dropwise, transparent reverse micro emulsion is obtained.0.75-3mLTEOS is added dropwise again,
Reacted 72 hours with 600 revs/min at room temperature.Add acetone demulsification, centrifugation to be washed through methyl alcohol, collect Fe3O4/SiO2Receive
Rice corpuscles；
Step 3：Prepare Fe3O4/SiO2/mSiO2Nano-particle
1-4g CTAC and Fe3O4/SiO2Nanoparticle dissolution is had children outside the state plan 30 minutes in 10-40mL water.Then, 41- is added
164 μ LTEA, 80 DEG C are reacted 60 minutes.0.75-3mL TEOS are added, is reacted 60 minutes with 600 revs/min at room temperature.Plus
Enter acetone demulsification, centrifugation is washed through methyl alcohol.1%NaCl methanol solutions are used again, are flowed back 3 hours at 60 DEG C, centrifugation,
It is repeated 5 times, removes CTAC, collects Fe3O4/SiO2/mSiO2Nano-particle；
Step 4：Prepare Fe3O4/SiO2/mSiO2-NH2Nano-particle
25-100mg Fe3O4/SiO2/mSiO2It is dissolved in 25-100mL ethanol, adds 50-200 μ L APTES stirrings 4
Hour, Fe is collected in centrifugation3O4/SiO2/mSiO2-NH2Nano-particle；
Step 5：Prepare Fe3O4/SiO2/mSiO2- (Gd-DTPA) nano-particles
25-100mg DTPA and Fe3O4/SiO2/mSiO2-NH2It is dissolved in 10-40mL water, 40 DEG C are stirred 3 hours, then are added
Enter 50-200mgGdCl3, stir 12 hours, wash with water, collect Fe3O4/SiO2/mSiO2- (Gd-DTPA) nano-particles.
Step 6：Prepare Fe3O4/SiO2/mSiO2- (Gd-DTPA)-PEG-RGE nano-particles
25-100mg RGERPPR-PEG-COOH and Fe3O4/SiO2/mSiO2- (Gd-DTPA) is dissolved in 10-40mL ethanol
In, 40 DEG C are stirred 3 hours, with ethanol and water washing, collect Fe3O4/SiO2/mSiO2- (Gd-DTPA)-PEG-RGE nanoparticles
Son.
Step 7：Prepare Fe3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE nano-particles
0.5-2mL solubility is the DOX PBS solutions and Fe of 0.33mg/mL3O4/SiO2/mSiO2-(Gd-DTPA)-PEG-
RGE, stirs 12 hours, collects Fe3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE nano-particles, the nano-particle is
The nanoscale medicine delivery system, its uv measurement DOX drugloading rates are 10-35%.
PEG molecular weight is 400-10000 in the RGERPPR-PEG-COOH.
The rotating speed of the centrifugation is 2000-12000 revs/min, 5-30 minutes.
The application of the nanoscale medicine delivery system, feature is that the delivery system is passed for the targeting of glioma treatment medicine
Send.
Following measure has been made respectively to nanoscale medicine delivery system of the invention：
Determine the particle diameter of the nanoscale medicine delivery system；
Determine the nanoscale medicine delivery system relaxation rate；
Determine intake of the nanoscale medicine delivery system to U87MG cells；
Determine Magnetic resonance imaging of the nanoscale medicine delivery system in glioma nude mouse；
Determine inhibitory action of the nanoscale medicine delivery system to glioma tumour growth.
Measurement result shows that nanoscale medicine delivery system of the invention can be used as tumor imaging diagnosis tracer drug, it is also possible to make
The integration of the delivering of antineoplastic, diagnosing tumor and oncotherapy.
Delivery system of the invention provides a kind of cancer target multifunctional nano of tumour penetrating peptide RGERPPR modifications and passs medicine
System, Fe3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE is used for the diagnosis and treatment integration of diagnosing tumor and oncotherapy.
Delivery system Fe of the invention3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE can be administered by being injected intravenously, by
RGERPPR mediations and tumour EPR effects kill tumour cell targeted to tumor tissues, in addition, it is by T1 contrast agent Gd-
DTPA and T2 contrast agent Fe3O4Utilize simultaneously, can by Magnetic resonance imaging spike Nano medication, there is provided pin-point accuracy is examined
Disconnected information, so as to realize the diagnosis and treatment integration of diagnosing tumor and oncotherapy.
Brief description of the drawings
Fig. 1 is Fe of the present invention3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE delivery system syntheti c route figures；
Fig. 2 is Fe of the present invention3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE transmission electron microscope pictures；
Fig. 3 is Fe of the present invention3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE relaxation rate lab diagrams；
Fig. 4 is Fe of the present invention3O4/SiO2/mSiO2The intake experiment of/DOX- (Gd-DTPA)-PEG-RGE cells qualitative, quantitative
Figure；
Fig. 5 is Fe of the present invention3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE Magnetic resonance imaging lab diagrams；
Fig. 6 is Fe of the present invention3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE animal efficacy experiment figures.
Specific embodiment
In order to be better understood from the present invention, below with embodiment come the present invention is furture elucidated, but present disclosure is not
It is limited only to example below.
Embodiment 1
Prepare Fe3O4Nano-particle
2.78g iron oleates add 0.48mL octadecylenes after 30 DEG C of vacuum drying chambers stand 24 hours.Mixture is heating up
3.3 DEG C of min of speed-1320 DEG C are heated to, reaction 60 minutes is then kept at this temperature.After reaction, room temperature is cooled to, passed through
10mL n-hexanes and the washing of 40mL acetone, centrifugation, obtain Fe3O4Nano-particle.
Embodiment 2
Prepare Fe3O4/SiO2Nano-particle
6.8mg Fe3O4Nano-particle and 8mL Igepal CO-520 are dissolved in 170mL hexamethylenes, are had children outside the state plan 30 minutes.
Then, 1.3mL ammoniacal liquor is added dropwise, transparent reverse micro emulsion is obtained.1.5mLTEOS is added dropwise again, at room temperature with 600 revs/min
Reaction 72 hours.Add acetone demulsification, centrifugation to be washed through methyl alcohol, collect Fe3O4/SiO2Nano-particle.
Embodiment 3
Prepare Fe3O4/SiO2/mSiO2Nano-particle
2g CTAC and Fe3O4/SiO2Nanoparticle dissolution is had children outside the state plan 30 minutes in 20mL water.Then, 82 μ LTEA are added,
80 DEG C are reacted 60 minutes.1.5mL TEOS are added, is reacted 60 minutes with 600 revs/min at room temperature.Acetone demulsification is added,
Centrifugation, washs through methyl alcohol.1%NaCl methanol solutions are used again, is flowed back 3 hours at 60 DEG C, centrifugation is repeated 5 times, remove
CTAC, collects Fe3O4/SiO2/mSiO2Nano-particle.
Embodiment 4
Prepare Fe3O4/SiO2/mSiO2-NH2Nano-particle
50mg Fe3O4/SiO2/mSiO2It is dissolved in 50mL ethanol, adds 100 μ L APTES to stir 4 hours, centrifugation point
From collection Fe3O4/SiO2/mSiO2-NH2Nano-particle.
Embodiment 5
Prepare Fe3O4/SiO2/mSiO2- (Gd-DTPA) nano-particles
50mg DTPA and Fe3O4/SiO2/mSiO2-NH2It is dissolved in 20mL water, 40 DEG C are stirred 3 hours, are added
100mgGdCl3, stir 12 hours, wash with water, collect Fe3O4/SiO2/mSiO2- (Gd-DTPA) nano-particles.
Embodiment 6
Prepare Fe3O4/SiO2/mSiO2- (Gd-DTPA)-PEG-RGE nano-particles
50mg RGERPPR-PEG-COOH and Fe3O4/SiO2/mSiO2- (Gd-DTPA) is dissolved in 20mL ethanol, 40 DEG C
Stirring 3 hours, with ethanol and water washing, collects Fe3O4/SiO2/mSiO2- (Gd-DTPA)-PEG-RGE nano-particles.
Embodiment 7
Prepare Fe3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE nano-particles
1mL solubility is the DOX PBS solutions and Fe of 0.33mg/mL3O4/SiO2/mSiO2- (Gd-DTPA)-PEG-RGE, stir
Mix 12 hours, collect Fe3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE nano-particles, uv measurement DOX drugloading rates
It is 27.2%.
Embodiment 8
By Fe3O4(Fig. 2A), Fe3O4/SiO2(Fig. 2 B), Fe3O4/SiO2/SiO2(Fig. 2 C) and Fe3O4/SiO2/mSiO2(figure
2D) form is observed by transmission electron microscope, specific as follows：(1) sample solution is dripped on copper mesh, in (2) air
Dry about 10 minutes, unnecessary liquid is sucked with filter paper, (3) are observed under transmission electron microscope is placed in after sample drying.Fig. 2 results show
Show the equal comparison rule of nanoparticle form, it is spherical in shape.
Embodiment 9
The T1 relaxation times and T2 relaxation times of probe under different Gd concentration are measured using 3T MRI.With Gd concentration and relaxation
The slope of the linearity curve of time 1/T1 calculates corresponding r1, with Fe concentration and the slope meter of the linearity curve of relaxation time 1/T2
Calculate corresponding r2.Fe3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE is diluted in distilled water, and Gd concentration ranges are
0.1-0.5mM.In transferring the sample into 96 orifice plates, and with the following parameter measurement T1 relaxation times：TR=7000ms, TE=
11ms, TI=24,100,200,400,600,900,1200,2000,3000 and 5000ms, FOV=120 × 85mm, average
=1.The following parameter measurement T2 relaxation times：TR=3000ms, TE=14.3ms, FOV=87x280mm, matrix=
240x768, slice thickness=5mm, average=1.Experimental result is as shown in figure 3, the r1 relaxation rates of Gd-DTPA
6.13mM-1S-1, Fe3O4R2 relaxation rates 36.89mM-1S-1。
Embodiment 10
Cell in vitro is absorbed：With U87MG cells as model, by cell with 3 × 104The density kind of cells/ml is in 24 orifice plates
Middle culture 24 hours, then with the Fe for being loaded with fluorescent dye DIO3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG and Fe3O4/
SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE drug-treateds 4h.Washed with PBS three times after agent-feeding treatment, laser is used respectively
The burnt qualitative and flow cytometer quantitative assessment cellular uptake situation of copolymerization.Need to use in nucleus with laser co-focusing qualitative analysis
Hochest dyes blueness.The cell of flow cytometry analysis is first washed three times with PBS, is washed with PBS, is digested, is centrifuged, will
Precipitate resuspended, flow cytomery.As shown in Figure 4 B, laser co-focusing result qualitative observation shows that tumour penetrating peptide is modified
Fe3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE is easily by cellular uptake.Flow cytometer quantitative expedition result
Display that, Fe3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE nano-particles are easily by cellular uptake.
Embodiment 11
Carry out MR imaging research.After nude inoculation U87MG cells two weeks, by intraperitoneal injection yellow Jackets (50 μ
L, 2.5%, 50mg/kg) mouse (n=3) is anaesthetized.MR images are obtained in the 3T MRI scanners with animal coil.It is logical
Cross tail vein and Fe is injected with the dosage of 0.07mmol-Gd/kg body weight3O4/SiO2/mSiO2/DOX-(Gd-DTPA)-PEG-RGE(C
T1 and D T2), and with the Fe of same dose3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG (A T1 and B T2) is used as right
According to.It is scanned in different time points.T1 sequential parameters for IMAQ are as follows：TR=400ms, TE=14ms, α=
90 °, FOV=45 (60,75%) mm, slice thickness=1mm, image matrix=256 × 256.The measurement ginseng of T2
Number is：TR=3500ms, TE=71ms, FOV=70 (70,100%) mm, matrix=256x256, slice thickness
=1.2mm.Experimental result is as shown in figure 5, Fe3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE had preferably at 8 hours
Contrasting effects, tumour penetrating peptide RGERPPR mediations and tumour EPR effects make Fe3O4/SiO2/mSiO2/DOX-(Gd-
DTPA)-PEG-RGE is enriched with tumor locus, for the diagnosis and treatment integration of diagnosing tumor and oncotherapy provides feasibility.
Embodiment 12
By U87MG cells routinely condition (37 DEG C, 5%CO2) culture, when cell fusion degree reaches 80-90%, with containing
0.25% trypsin digestion cell of EDTA is simultaneously collected by centrifugation, and is counted twice with the PBS washed cells of PH 7.4, is prepared into final concentration
It is 5 × 106The single cell suspension of individual/mL, the right shoulder for seeding cells into nude mice with 1mL syringes is subcutaneous, every nude inoculation
0.1mL, when tumour is long to 100-120mm3When it is standby.When tumour is long to 100-120mm3When (be denoted as 0 day), by tumor-bearing mice with
Machine is divided into 4 groups, every group 6, is set to saline control group, DOX (5mg/kg) group, Fe3O4/SiO2/mSiO2/DOX-
(Gd-DTPA)-PEG (5mg/kg) group, Fe3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE (5mg/kg) group, tail is quiet
Arteries and veins drug administration by injection, investigates following index：
Tumor growth curve：The every two days major diameters and minor axis with vernier caliper measurement tumour, it is (long according to formula v=1/2
Footpath * minor axis2), the volume of tumour is calculated, draw the growth curve that gross tumor volume is changed over time.
Efficacy experiment result as shown in fig. 6, compared with saline control group, Fe3O4/SiO2/mSiO2/DOX-(Gd-
DTPA)-PEG-RGE groups show significantly to suppress the effect of tumour growth.
Claims (5)
1. a kind of cancer target multifunctional nano delivery system, it is characterised in that the delivery system includes tumour penetrating peptide
The multifunctional nano grain and antineoplastic of RGERPPR modifications, the multifunctional nano grain is T1-T2 bimodulus magnetic resonance radiographies
Agent；The antineoplastic is doxorubicin or epirubicin；Percentage by weight is 10%- to its antineoplastic in systems
35%；Its structure is as follows：
Wherein：Delivery system is nanoparticle, and particle diameter is 0.10-0.60 in 60-200nm, particle diameter distribution PDI, and n is the degree of polymerization, n=
9-227。
2. the preparation method of nanoscale medicine delivery system described in a kind of claim 1, it is characterised in that the method includes walking in detail below
Suddenly：
Step 1：Prepare Fe3O4Nano-particle
1.39-5.56g iron oleates add 0.24-0.96mL octadecylenes after 30 DEG C of vacuum drying chambers stand 24 hours；Mixture
With 3.3 DEG C of min of heating rate-1320 DEG C are heated to, reaction 60 minutes is then kept at this temperature；After reaction, room is cooled to
Temperature, through 5-20mL n-hexanes and the washing of 20-80mL acetone, centrifugation, obtains Fe3O4Nano-particle；
Step 2：Prepare Fe3O4/SiO2Nano-particle
3.4-13.6mg Fe3O4Nano-particle and 4-16mL Igepal CO-520 are dissolved in 85-340mL hexamethylenes, excusing from death
30 minutes；Then, 0.65-2.6mL ammoniacal liquor is added dropwise, transparent reverse micro emulsion is obtained；0.75-3mLTEOS is added dropwise again, in room temperature
Under with 600 revs/min react 72 hours；Add acetone demulsification, centrifugation to be washed through methyl alcohol, collect Fe3O4/SiO2Nanoparticle
Son；
Step 3：Prepare Fe3O4/SiO2/mSiO2Nano-particle
1-4g CTAC and Fe3O4/SiO2Nanoparticle dissolution is had children outside the state plan 30 minutes in 10-40mL water；Then, 41-164 μ are added
LTEA, 80 DEG C are reacted 60 minutes；0.75-3mL TEOS are added, is reacted 60 minutes with 600 revs/min at room temperature；Add third
Ketone is demulsified, centrifugation, is washed through methyl alcohol；1%NaCl methanol solutions being used again, being flowed back 3 hours at 60 DEG C, centrifugation repeats 5
It is secondary, CTAC is removed, collect Fe3O4/SiO2/mSiO2Nano-particle；
Step 4：Prepare Fe3O4/SiO2/mSiO2-NH2Nano-particle
25-100mg Fe3O4/SiO2/mSiO2It is dissolved in 25-100mL ethanol, adds 50-200 μ L APTES to stir 4 hours,
Centrifugation, collects Fe3O4/SiO2/mSiO2-NH2Nano-particle；
Step 5：Prepare Fe3O4/SiO2/mSiO2- (Gd-DTPA) nano-particles
25-100mg DTPA and Fe3O4/SiO2/mSiO2-NH2It is dissolved in 10-40mL water, 40 DEG C are stirred 3 hours, are added
50-200mgGdCl3, stir 12 hours, wash with water, collect Fe3O4/SiO2/mSiO2- (Gd-DTPA) nano-particles；Step 6：
Prepare Fe3O4/SiO2/mSiO2- (Gd-DTPA)-PEG-RGE nano-particles
25-100mg RGERPPR-PEG-COOH and Fe3O4/SiO2/mSiO2- (Gd-DTPA) is dissolved in 10-40mL ethanol, and 40
DEG C stirring 3 hours, with ethanol and water washing, collects Fe3O4/SiO2/mSiO2- (Gd-DTPA)-PEG-RGE nano-particles；Step
7：Prepare Fe3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE nano-particles
0.5-2mL solubility is the DOX PBS solutions and Fe of 0.33mg/mL3O4/SiO2/mSiO2- (Gd-DTPA)-PEG-RGE, stir
Mix 12 hours, collect Fe3O4/SiO2/mSiO2/ DOX- (Gd-DTPA)-PEG-RGE nano-particles, the nano-particle is received for described
Rice delivery system, its uv measurement DOX drugloading rates are 10-35%.
3. preparation method according to claim 2, it is characterised in that PEG molecular weight is in the RGERPPR-PEG-COOH
400-10000。
4. preparation method according to claim 2, it is characterised in that the rotating speed of the centrifugation be 2000-12000 turn/
Minute, 5-30 minutes.
5. the application of nanoscale medicine delivery system described in a kind of claim 1, it is characterised in that the delivery system is controlled for glioma
Treat the targeted delivery of medicine.
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