CN105983106A - IL-13 modified gadolinium chelate containing liposome targeted magnetic resonance imaging contrast agent and preparation method and application thereof - Google Patents
IL-13 modified gadolinium chelate containing liposome targeted magnetic resonance imaging contrast agent and preparation method and application thereof Download PDFInfo
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- CN105983106A CN105983106A CN201510319595.1A CN201510319595A CN105983106A CN 105983106 A CN105983106 A CN 105983106A CN 201510319595 A CN201510319595 A CN 201510319595A CN 105983106 A CN105983106 A CN 105983106A
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- liposome
- gadolinium chelate
- chelate compound
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- 239000013522 chelant Substances 0.000 title claims abstract description 48
- 239000002405 nuclear magnetic resonance imaging agent Substances 0.000 title claims abstract description 30
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 150000000921 Gadolinium Chemical class 0.000 title abstract 3
- 229910052688 Gadolinium Inorganic materials 0.000 claims abstract description 56
- UIWYJDYFSGRHKR-UHFFFAOYSA-N gadolinium atom Chemical group [Gd] UIWYJDYFSGRHKR-UHFFFAOYSA-N 0.000 claims abstract description 55
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Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Magnetic Resonance Imaging Apparatus (AREA)
Abstract
The invention discloses an IL-13 modified gadolinium chelate containing liposome targeted magnetic resonance imaging contrast agent and a preparation method and application thereof. The contrast agent is gadolinium chelate encapsulated liposome, and the surface of the liposome is modified by IL-13. The preparation method of the IL-13 modified gadolinium chelate containing liposome targeted magnetic resonance imaging contrast agent comprises the steps that a liposome film is firstly prepared; then gadolinium chelate is added into a container containing the liposome film; the gadolinium chelate is wrapped by the liposome film with the assistance of ultrasonic waves to form gadolinium chelate containing multilamelar liposome; the surface of the gadolinium chelate encapsulated multilamelar liposome is modified by thiolated IL-13. The contrast agent can penetrate through a blood brain barrier to be transferred to a tumor diseased region and be specifically bound with the tumor diseased region. Therefore, the contrast agent has targeting ability, can detect a tumor focus smaller than 2 mm and can be used for early detection of tumors.
Description
Technical field
The invention belongs to medical imaging technology field, be specifically related to the lipid containing gadolinium chelate compound that IL-13 modifies
Body targeting magnetic resonance imaging contrast agent and its preparation method and application.
Background technology
The technology being presently used for diagnosing tumor mainly includes computed tomography (CT), positron emission meter
Calculation machine tomoscan (PET), optical imagery and ultrasonic imaging etc..Because of magnetic resonance imaging (MRI) not ionization
Radiation, and by contrast medium, the characteristics such as accurate positioning, quantitative analysis can be carried out to deep tissue, become
For one of detection conventional clinically and diagnostic means.
Intestinal cancer, also referred to as colorectal cancer, it occurs at colon or rectum, belongs in the U.S. and diagnoses the 3rd arriving
Plant most commonly seen cancer, and belong to the third-largest cancer of Cancer death.Its harmfulness is, once intestines in late period
During carcinogenesis hepatic metastases, partially hepatectomized only has 25% survival rate, and recurrence rate but has 75%, and this is 75% again
Send out rate great majority and ultimately result in death, in this 75% recurrence rate, there are about the transfer of 50% can transfer to liver with
Outer tissue.IL-13 alpha-2 receptor height is had to express in pernicious intestinal cancer transfer cell, and in the transfer of intestinal cancer
Adhesion in journey, migrating, living in, metastatic moves to be grown period and plays an important role, and causes late period
Intestinal cancer prognosis is difficult to ideal.
Adrenocortical carcinoma is relatively rare, but it be a kind of there is aggressive and temporarily without the evil of effective treatment method
Property tumour.Result of study display adrenocortical carcinoma has IL-13 alpha-2 receptor height expresses, and IL-13 α 2 plays
Regulate pernicious adrenocortical carcinoma tumour cell to live in, regenerate breeding.Adrenocortical carcinoma IL-13 alpha-2 receptor
Highly express the novel targeting target being to detect with chemotherapy adrenocortical carcinoma.
Glioma is a kind of primary malignant tumor being grown in central nervous system, and brain tissue is had by it
Very strong aggressive, and astrocytoma, mesoglioma, ependymoma, medulloblastoma can be subdivided into
And glioblastoma multiforme.The conventional treatment regime of glioma is that Post operation adds radiotherapy or chemotherapy,
Unfortunately be the poor prognosis of conventional treatment regime, the average survival time rate only about 14 months.Cause so low
The one of the main reasons of survival rate is that we lack a kind of sensitive detection means in early days, conventional detection hand at present
The focus less than 2mm for the Duan Wufa detection, when the cancer of the brain is less than 2mm, conventional detection means are malfunctioning, and this is
By the machine extensively applying magnetic resonance imaging contrast agent (containing gadolinium chelate compound such as Magnevist Solution etc.) clinically at present
System is determined.Magnetic resonance Magnevist Solution is only capable of the cancer detecting more than more than 2mm, and it is often
Belong to advanced tumor.Advanced tumor blood-brain barrier is impaired, and Magnevist Solution can be by impaired blood-brain barrier
And enter the growth site of tumour and accumulate on this in a large number, make NMR can capture Magnevist Solution institute
The signal intensity producing.And during early lesion when being limited in that tumour is less than 2mm of Magnevist Solution
Blood-brain barrier is relatively complete, and Magnevist Solution is limited to blood-brain barrier thus cannot accumulate on tumor focus position,
Thus Magnetic resonance imaging desired signal cannot be provided to change, cause NMR to detect swollen in early days
Knurl and wellability focus.Meanwhile, conventional contrast agents can not produce specific binding with tumour cell, i.e. belongs to non-
Target contrast medium, it is impossible to accurately distinguish lesions position and normal structure, causes the focus cannot be by when operation
Excising completely, recurrence frequency is raw then.Therefore, the wellability focus diagnosis of tumour early stage, especially less than 2mm
Effective detection of following cancer lesions position, all plays key effect to raising oncotherapy effect or tumor eradication.
But conventional MRI imaging (MRI) is even if using contrast medium brain tumor in early days also cannot be detected.Cause
This, exploitation one can pass through blood-brain barrier and is combined with tumor tissue specificity, to detect early stage aggressive swollen
The novel contrast medium of tumor tissue has very important clinical meaning.
Many tumours express interleukin-13 alpha-2 receptor (IL-13R α 2) at its cell surface height, for example, go up
The glioma stated all has IL-13 α 2 height with the liver cancer etc. that carcinoma of the rectum transfer is gone expresses.
In order to improve the targeting of gadolinium (Gd) contrast preparation, researcher utilizes tumor cell surface IL-13R α 2 high
The characteristic that degree is expressed, allows tumor cell surface IL-13R α 2 combine with ligand IL-13 being loaded with medicine.
After the IL-13R α 2 on tumour cell occurs specific binding with ligand IL-13 being loaded with medicine, lipid
The contained carrier gadolinium of body is delivered to the tumour cell position that acceptor IL-13R α 2 height is expressed, thus reaches mark
The effect of target.
Chinese patent 201110312974.X discloses a kind of close based on biotinylated mAb and strepto-
Combine the targeting glioma magnetic resonance contrast that bag carries two kinds of compounds of sterically stabilized immunoliposomes of gadolinium with element
Agent；Described monoclonal antibody is Endoglin (CD105), for the detection of glioma new vessels.This is right
Inject in two steps when using than agent, i.e. need first to give biotinylated mAb, then give Streptavidin
Carry the sterically stabilized immunoliposomes of gadolinium in conjunction with bag.
Chinese patent 201410252588.X discloses a kind of fat being carried gadolinium by aptamer analog and bag
The magnetic resonance imaging contrast agent that plastid is constituted, described aptamer analog selected from mercapto is modified, amino
Modify or site modify GBI-10, AS1411, A10, MUC1, sgc8, sgc3, sgd3, KH1C1,
One or more in KD05, S6, AGR0100, A30 and STRG2, are used for early detection tumour
Cell, wherein GBI-10 can be used for the diagnosis of glioma.But in liposome preparation process, extruded film
Up to 20 times.
Chinese patent 201010286508.4 discloses a kind of with polypeptide for targeting head base, dendrimer material
Macromolecule carrier based on material, surface connects little molecule magnetic resonance imaging contrast agent, makes cancer target magnetic altogether
Shake imaging contrast, i.e. utilize and peptide T 7 peptide of specific binding tumor cell surface Tf acceptor can modify height
Molecular material, connects magnetic resonance imaging contrast agent and makes cancer target magnetic resonance contrast agent.
Chinese patent 200910196477.0 discloses a kind of nano liposomes and the gadolinium pair of target polypeptide modification
Ratio agent, reaches tumour identification and contrast preparation delays, and target polypeptide is modified and is selected from GD10 peptide molecule, F56
Polypeptide, K237 peptide molecule.Described contrast medium particle diameter is at 20～200nm, size distribution width.
Chinese patent 201080026113.9 discloses a kind of pulullan polysaccharide and DTPA (diethylenetriamines five
Acetic acid dianhydride) in the pulullan polysaccharide-DTPA conjugant that formed by ester bond gadolinium coordination be incorporated into DTPA and shape
The gadolinium contrast preparation becoming, is used for liver MRI image.
Content of the invention
In order to improve accuracy of detection, solve the problems referred to above of the prior art, the invention provides a kind of IL-13
The targeting magnetic resonance imaging contrast agent of liposome containing gadolinium chelate compound modified.
The targeting magnetic resonance imaging contrast agent of liposome containing gadolinium chelate compound that the IL-13 that the present invention provides modifies, be
The liposome of encapsulating gadolinium chelate compound, and surface of liposome is through IL-13 modification.
Interleukins IL-13 has modified surface of liposome, and liposome encloses gadolinium chelate compound.White thin
Born of the same parents' interleukin IL-13 has height homogenieity, the interleukin-13 R α 2 that can over-express with lesion
Receptor-specific combines.For example, for malignant brain tumor, each malignant brain tumor cell there are about
16000～22000 IL-13R alpha-2 receptor binding sites, therefore height is expressed white by this magnetic resonance imaging contrast agent
The cancer cell of cytokine 13R alpha-2 receptor has very strong affinity.
Preferably, in the targeting magnetic resonance imaging contrast agent of liposome containing gadolinium chelate compound that above-mentioned IL-13 modifies,
Described gadolinium chelate compound is Magnevist Solution or gadoterlc acid meglumine saltlniection.
Magnevist Solution (Gd-DATP), has another name called the double meglumine of diethylene triamine pentaacetic acid gadolinium, is presently the most
Widely used contrast medium.Gadoterlc acid meglumine saltlniection (Gd-DOTA) is also the more commonly used gadolinium contrast medium, equally
It is applicable to the present invention.
Preferably, in the targeting magnetic resonance imaging contrast agent of liposome containing gadolinium chelate compound that above-mentioned IL-13 modifies,
The raw material of preparing of described liposome includes DPPC (DPPC), distearyl acyl group phosphatidyl second
Hydramine-polyethylene glycol (DSPE-PEG), cholesterol (CHOL) and DSPE-poly-second two
Alcohol-maleimide cross-linking agent (DSPE-PEG-Mal).The mixing of this different materials can make multilamelar liposome,
Can effectively encapsulate gadolinium chelate compound, and pass through blood-brain barrier arrival focus.
Preferably, in the targeting magnetic resonance imaging contrast agent of liposome containing gadolinium chelate compound that above-mentioned IL-13 modifies,
DPPC, DSPE-PEG, cholesterol and distearyl
The mol ratio of base phosphatidyl-ethanolamine-polyethylene glycol-maleimide cross-linking agent is 10:5:5:0.5.This consumption
Liposome prepared by the material of proportioning, envelop rate is high, yield rate is high, and is used to suppress brain colloid carcinoma curative effect
Preferable.
The present invention also provides more than one targeting magnetic of liposome containing gadolinium chelate compound that arbitrary described IL-13 modifies
The preparation method of resonance image-forming contrast medium, step is as follows:
(1) by DPPC, DSPE-PEG, cholesterol
It is dissolved in volume ratio 1:1 with DSPE-PEG-maleimide cross-linking agent
In methanol chloroform mixed solvent, under 40 DEG C of rotatory vacuum reduced pressure, solvent is volatilized and is formed film, film
It is dried 10～20min in nitrogen, be then mothballed 14～16h in 4 DEG C of driers；Gadolinium chelate compound is added into
In the container containing liposome membrane, under the assistance of ultrasonic wave, gadolinium chelate compound is wrapped up by liposome membrane
To form the multilamelar liposome containing gadolinium chelate compound；This multilamelar liposome under the conditions of-80 DEG C by freeze thawing four times, so
It is extruded through the polycarbonate membrane four times of aperture 200nm afterwards by nitrogen, then be extruded through aperture by nitrogen
The polycarbonate membrane of 100nm four times；
(2) IL-13 is dissolved in the phosphate buffer containing EDTA, adds final concentration 1.4mM's
The 2 Iminothiolane hydrochloride aqueous solution, then under the conditions of 4 DEG C, stirring makes IL-13 Thiolation；Will be through mercaptan
The IL-13 solution changed is purified through G25 cross-linked dextran gel column, and purified product is concentrated to
1mL, then be added in step (1) products therefrom, under the conditions of 4 DEG C, stir 12h, obtain IL-13
The targeting magnetic resonance imaging contrast agent of liposome containing gadolinium chelate compound modified.
What the Thiolation process of IL-13 protein produced is Thiolation IL-13 and is not freely dissociated by Thiolation
The mixture of IL-13, not removed IL-13 not Thiolation freely, cross G25 sephadex
Post is requisite operation.
Preferably, the preparation of the targeting magnetic resonance imaging contrast agent of liposome containing gadolinium chelate compound that above-mentioned IL-13 modifies
Method, also includes purification step (3): the liposome containing gadolinium chelate compound that the IL-13 that step (2) obtains modifies
Targeting magnetic resonance imaging contrast agent first cross agarose CL-2B pillar to remove unconjugated Thiolation IL-13,
Then dialyse to exchange solvent in 4 DEG C of phosphate buffers；A phosphate buffer is changed after dialysis 4h,
Total dialysis time is 16～20h；Then, centrifugal concentrating under the conditions of 4 DEG C, the IL-13 obtaining purifying modifies
Liposome containing gadolinium chelate compound targeting magnetic resonance imaging contrast agent.
The targeting magnetic resonance imaging contrast agent of liposome containing gadolinium chelate compound that the above-mentioned IL-13 of the present invention modifies is in inspection
Surveying the application in tumour, IL-13 α 2 expression of described tumour is higher than Healthy People normal value (such as brain colloid
Knurl, pernicious intestinal cancer shifts, adrenocortical carcinoma).
Preferably, described tumour is glioma, intestinal cancer and/or adrenocortical carcinoma.
Compared with prior art, the method have the advantages that
The targeting magnetic resonance imaging contrast agent of liposome containing gadolinium chelate compound that 1.IL-13 modifies can assist contrast medium
Through blood-brain barrier；
The magnetic that bag can be carried by the targeting magnetic resonance imaging contrast agent of liposome containing gadolinium chelate compound that 2.IL-13 modifies is altogether
Image contrast of shaking agent is delivered to lesion and produces specific binding with cancer cell, possesses targeting；
The targeting magnetic resonance imaging contrast agent of liposome containing gadolinium chelate compound that 3.IL-13 modifies can detect and be less than
2mm tumor focus, can be used for the early stage detection of tumour.
Brief description
Fig. 1 is IL-13-Liposomes-Gd-DTPA stereoscan photograph.
Fig. 2 is nude mice intravenous injection IL-13-Liposomes-Gd-DTPA MRI Detection results.
Fig. 3 is nude mice intravenous injection Gd-DTPA MRI Detection results.
Fig. 4 is nude mice intravenous injection IL-13-Liposomes-Gd-DTPA Brain slices MRI Detection results.
Fig. 5 is nude mice intravenous injection Gd-DTPA Brain slices MRI Detection results.
Detailed description of the invention
Below in conjunction with specific embodiment, the invention will be further described, makes those skilled in the art can be more
The good understanding present invention simultaneously can be practiced, but illustrated embodiment is not as a limitation of the invention.
The preparation of embodiment 1 IL-13-liposomes-Gd-DTPA.
By DPPC 80mg, DSPE-PEG 13mg,
Cholesterol 20mg and DSPE-PEG-maleimide cross-linking agent 10mg (rubs
That ratio is 10:5:5:0.5) it is dissolved in 40mL methanol chloroform (the two volume ratio is 1:1) mixed solvent, 40 DEG C
Under rotatory vacuum reduced pressure, solvent is volatilized and is formed film, and film is dried 10～20min in nitrogen, so
After be mothballed 14～16h in 4 DEG C of driers.Magnevist Solution is added into the container containing liposome membrane
In, under the assistance of ultrasonic wave, liposome membrane wraps up magnetic resonance imaging contrast agent to be formed containing magnetic resonance
The multilamelar liposome of imaging contrast.This liposome by freeze thawing four times, is then squeezed by nitrogen under the conditions of-80 DEG C
Pressure is through the polycarbonate membrane four times of aperture 200nm, then is extruded through the poly-carbonic acid of aperture 100nm by nitrogen
Ester film four times.
IL-13 protein 2mg is dissolved in 6mL and contains 0.67mM disodium ethylene diamine tetraacetate (EDTA)
Phosphate buffer in, add the 2 Iminothiolane hydrochloride aqueous solution that ultimate density is 1.4mM,
Then under the conditions of 4 DEG C, stir 30min, make IL-13 protein Thiolation.IL-13 albumen after Thiolation
Matter obtains bright sulfur alcoholization by G-25 cross-linked dextran gel column to be isolatable from by free IL-13
IL-13, the IL-13 eluent of bright sulfur alcoholization is concentrated into 1mL.Thiolation IL-13 is joined containing gadolinium spray
In the liposome of acid Portugal amine, under the conditions of 4 DEG C, stir 12h, obtain liposomes vs's agent that IL-13 modifies.
It is uncombined to remove that agarose CL-2B pillar is first crossed in liposomes vs's agent that prepared IL-13 modifies
Thiolation IL-13, then dialyse to exchange solvent in 4 DEG C of phosphate buffers.Change after dialysis 4h
Phosphate buffer, total dialysis time is 16～20h.Then, at 4 DEG C of centrifugal concentratings to 8.0mL,
Obtain, named IL-13-liposomes-Gd-DTPA.
Prepared IL-13-liposomes-Gd-DTPA is spherical in shape, and size dimension is 80～140nm, averagely
A size of 131nm, liposome polydispersity index (PDI) is 0.1～0.2nm, sees Fig. 1.
Embodiment 2 IL-13-liposomes-Gd-DTPA may pass through blood-brain barrier
The targeting magnetic resonance imaging contrast agent of liposome containing gadolinium chelate compound modified for the IL-13 proving
IL-13-liposomes-Gd-DTPA may pass through blood-brain barrier, we select normally, the nude mice without the cancer of the brain be
Model is tested.Result shows, the nude mice injection without the cancer of the brain gives magnevist contrast medium
(Magnevist), after Gd-DTPA 30min, nude mice pituitary signal noise ratio (signal to noise ratio) improves 25%,
On the contrary, brain essence signal to noise ratio but keeps constant.Show that magnevist contrast medium cannot pass through the blood brain of the nude mice that lives
Barrier is to reach brain parenchyma section.By contrast, the nude mice injection without the cancer of the brain gives
After IL-13-liposomes-Gd-DTPA 30min, not only the signal to noise ratio at nude mice pituitary position adds 20%,
Meanwhile, brain essence signal to noise ratio too increases 15%.This is due to IL-13-liposomes-Gd-DTPA
May pass through live body nude mouse blood-brain barrier, and penetrate into brain parenchyma region.Prove liposome as medicine
Thing carrier can be assisted contrast medium to pass through complete blood-brain barrier and be reached brain essence brain tissue.
The targeting of embodiment 3 IL-13-liposomes-Gd-DTPA
The liposome mr contrast agent modified by fluorescein isothiocynate cyano group mark IL-13
(FITC-IL-13-liposomes-Gd-DTPA) with without IL-13 liposome contrast agent
(FITC-liposomes-Gd-DTPA), in order at glioma U251 and GSC T3691 cell cytosol
Middle observation of cell absorbs the effect of contrast medium, and we have done cell absorption experiment.Tissue of biological cells result
Show, in colloid carcinoma stem cell GSC T3691 endochylema, observe that cell absorbs more IL-13 and modifies
FITC-IL-13-liposomes-Gd-DTPA.In its prompting IL-13 and GSC T3691 cell
IL-13R alpha-2 receptor has the compatibility of height, can farthest promote specific tumors cell pair
The absorption of IL-13-liposomes-Gd-DTPA.
The effect of embodiment 4 IL-13-lipsomes-Gd-DTPA detection tumour cell
It for proving that the IL-13-liposomes-Gd-DTPA that IL-13 modifies can detect that immature glioma, is
This carries out Related Experimental Study, and normal nude mice is vaccinated glioma U251 cell, after surrounding, nude mice brain
Inside grow maturation and early stage glioma, in this, as glioma animal model in brain.Give containing
Glioma nude mice injection magnevist contrast medium (Magnevist) Gd-DTPA in brain, by weighting 7T nuclear-magnetism altogether
Vibration Meter obtains image, sees Fig. 3.Same nude mice injection IL-13-liposomes-Gd-DTPA is given after 24h,
Obtain image by weighting 7T NMR, see Fig. 2.Result shows, IL-13-liposomes-Gd-DTPA
Can detect beyond nude mice maturity period focus, the wellability glioma of minimal disease also can be revealed,
Magnevist contrast medium then only show the ripe tumour in brain position, because now blood-brain barrier is broken
Bad, thus mr contrast agent can to enter brain tissue thin to show the brain tissue being destroyed by pathology rather than tumour
Born of the same parents' tissue itself.
In order to further determine that testing result, to intravenous injection Gd-DTPA (Magnevist) and
Nude mice after IL-13-liposomes-Gd-DTPA carries out the detection of brain deeper hierarchical slicing.Profound section knot
Fruit display, after intravenous injection Gd-DTPA, nude mice minimal disease cannot show, sees Fig. 5；Intravenous injection
The immature cancer of the brain of nude mice same brain position wellability of IL-13-liposomes-Gd-DTPA is clearly shown, sees
Fig. 4.Therefore, highly sensitive IL-13-liposomes-Gd-DTPA contrast medium for tumour early stage detection,
Early treatment is most important.
Embodiment described above is only the preferred embodiment lifted by absolutely proving the present invention, the present invention's
Protection domain is not limited to this.Those skilled in the art made on the basis of the present invention equivalent replacement or
Conversion, all within protection scope of the present invention.Protection scope of the present invention is as the criterion with claims.
Claims (8)
1. the targeting magnetic resonance imaging contrast agent of liposome containing gadolinium chelate compound that IL-13 modifies, its feature exists
In, for encapsulating gadolinium chelate compound liposome, and surface of liposome through IL-13 modify.
2. the targeting magnetic resonance imaging of liposome containing gadolinium chelate compound that IL-13 according to claim 1 modifies
Contrast medium, it is characterised in that described gadolinium chelate compound is Magnevist Solution or gadoterlc acid meglumine saltlniection.
3. the targeting magnetic resonance of liposome containing gadolinium chelate compound that IL-13 according to claim 1 and 2 modifies
Imaging contrast, it is characterised in that described liposome prepare raw material include DPPC,
DSPE-PEG, cholesterol and DSPE-poly-second
Glycol-maleimide cross-linking agent.
4. the targeting magnetic resonance imaging of liposome containing gadolinium chelate compound that IL-13 according to claim 3 modifies
Contrast medium, it is characterised in that DPPC, DSPE-poly-second two
The mol ratio of alcohol, cholesterol and DSPE-PEG-maleimide cross-linking agent
For 10:5:5:0.5.
5. the targeting magnetic of liposome containing gadolinium chelate compound that the arbitrary described IL-13 of a Claims 1 to 4 modifies is common
The preparation method of imaging contrast of shaking, it is characterised in that step is as follows:
(1) by DPPC, DSPE-PEG, cholesterol
It is dissolved in volume ratio 1:1 with DSPE-PEG-maleimide cross-linking agent
In methanol chloroform mixed solvent, under 40 DEG C of rotatory vacuum reduced pressure, solvent is volatilized and is formed film, film
It is dried 10～20min in nitrogen, be then mothballed 14～16h in 4 DEG C of driers；Gadolinium chelate compound is added into
In the container containing liposome membrane, under the assistance of ultrasonic wave, gadolinium chelate compound is wrapped up by liposome membrane
To form the multilamelar liposome containing gadolinium chelate compound；This multilamelar liposome under the conditions of-80 DEG C by freeze thawing four times, so
It is extruded through the polycarbonate membrane four times of aperture 200nm afterwards by nitrogen, then be extruded through aperture by nitrogen
The polycarbonate membrane of 100nm four times；
(2) IL-13 is dissolved in the phosphate buffer containing EDTA, adds final concentration 1.4mM's
The 2 Iminothiolane hydrochloride aqueous solution, then under the conditions of 4 DEG C, stirring makes IL-13 Thiolation；Will be through mercaptan
The IL-13 solution changed is purified through G25 cross-linked dextran gel column, and purified product is concentrated to
1mL, then be added in step (1) products therefrom, under the conditions of 4 DEG C, stir 12h, obtain IL-13
The targeting magnetic resonance imaging contrast agent of liposome containing gadolinium chelate compound modified.
6. the targeting magnetic resonance imaging of liposome containing gadolinium chelate compound that IL-13 according to claim 5 modifies
The preparation method of contrast medium, it is characterised in that also include purification step (3): the IL-13 that step (2) obtains
The targeting magnetic resonance imaging contrast agent of liposome containing gadolinium chelate compound modified first crosses agarose CL-2B pillar to remove
Unconjugated Thiolation IL-13, then dialyses to exchange solvent in 4 DEG C of phosphate buffers；Dialysis 4h
After change a phosphate buffer, total dialysis time is 16～20h；Then, centrifugal concentrating under the conditions of 4 DEG C,
Obtain the targeting magnetic resonance imaging contrast agent of liposome containing gadolinium chelate compound that the IL-13 of purifying modifies.
7. the targeting magnetic resonance of liposome containing gadolinium chelate compound that the arbitrary described IL-13 of Claims 1 to 4 modifies becomes
As application in detection tumour for the contrast medium, IL-13 α 2 expression of described tumour is higher than Healthy People normal value.
8. application according to claim 7, it is characterised in that described tumour is glioma, intestinal cancer
And/or adrenocortical carcinoma.
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