CN110354281A - A kind of pair of multi-modal molecular image probe of targeting and its preparation method and application - Google Patents
A kind of pair of multi-modal molecular image probe of targeting and its preparation method and application Download PDFInfo
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- CN110354281A CN110354281A CN201910537283.6A CN201910537283A CN110354281A CN 110354281 A CN110354281 A CN 110354281A CN 201910537283 A CN201910537283 A CN 201910537283A CN 110354281 A CN110354281 A CN 110354281A
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Abstract
The invention discloses a kind of double multi-modal molecular image probes of targeting and its preparation method and application.Probe of the invention is the double targeted probes of three mode for having nuclear-magnetism, optoacoustic, near-infrared fluorescent signal, using the strategy of the exclusive bis- targetings of EGFR and SEC61G, navigation application in its precision that can significantly improve preoperative diagnosing tumor and potential art, has broad application prospects and conversion value.
Description
Technical field
The invention belongs to molecular image probe technical fields, are related to a kind of pair of multi-modal molecular image probe of targeting and its system
Preparation Method and application.
Background technique
Glioma is the most common malignant primary brain tumor of central nervous system.According to brain tumor registration center, the U.S.
(CBTRUS) primary brain tumors and other central nerve neuroma diagnosed cases that 2010 to 2014 years of publication are collected
Statistical report show: wherein having the brain tumor case close to half (47.1%) is all patients with gliomas；And it is being diagnosed as glue
In the patient of matter tumor, has close to sixty percent (56.1%) and be diagnosed as the highest glioblastoma of grade malignancy (GBM).Due to
Glioma tumor cell can invade neighbouring normal cerebral tissue, therefore performed the operation along the nature of white matter fiber tract infiltration growth
Cheng Dangzhong is very difficult to identify small residual tumor and cannot thoroughly remove tumour cell, cause after performing the operation not
Evitable tumor recurrence, thus its prognosis is often very poor.Operation excision is the main means of the treatment GBM of current clinical use
With foundation stone treat, not only can rapidly reduction of patient partial clinical symptom, but also perform the operation for tumor resection complete journey
It spends closely bound up with the prognosis of patient.One III large-scale clinical trial phase is studied and confirmed: the degree of ocal resection is higher,
Patient's prognosis is better, and progression free survival phase and Overall survival significantly extend.But it due to the characteristic of glioma infiltration growth, invades
Violate normal adjacent tissue and structure, therefore the reality either in the preoperative diagnosis imaging of image department doctor or surgeon's art
When observation or " perception ", be all difficult to accurately define real tumor boundaries, and the excision that multirow is empirical.The consequence done so
Be exactly: on the one hand, excision not exclusively causes Neoplasm residual to easily lead to tumor recurrence；On the other hand, excessively excision causes important blood
Pipe and the probability of function core group damage increase, and lead to the damage of nervous function, influence patients ' life quality and level.Therefore, it " looks for
To " or " seeing " GBM patient tumors infiltration real border where, promoted operation excision precise degrees, for save patient
Critical function and extend patient's prognosis, have important value and significance.
Molecular image (molecularimaging) is a kind of based on cell or tissue specific target target label imaging skill
Art, by the instrumentality of molecular probe, the complicated physiological disease that we can be needed to observe on cell or molecular level
It manages phenomenon and carries out intuitive visual or reflection indirectly.Compared to traditional anatomy imaging, the feature of molecular image maximum is exactly can
In a manner of through Noninvasive, the change of molecular level under a certain physiology of reflection or pathological state of real-time quantitative, and often
This variation occurs before the pathological change of institutional framework, thus its diagnosing image of tumour early stage, clinical treatment decision,
Medicament research and development or even accurate medical treatment etc., which suffer from, to be widely applied.It is wide in clinical practice and pre- clinical level at present
The general image mode for using and studying has CT, MRI, PET, SPECT, US, optical molecular image and photoacoustic imaging etc..Different
Image mode, based on different image-forming principles, the sensitivity of diagnosis, resolution ratio, imaging depth, cost and in terms of
All respectively have superiority and inferiority, and the diagnostic message provided often gives priority to and tends to be single.Therefore, when the letter that a certain image mode provides
It ceases and is worth in limited situation, we will generate limitation or even deviation for the understanding of physiology or pathological phenomenon.And lead to
The chief for integrating a variety of image modes is crossed, the available various dimensions comprising molecule, function and anatomical information etc. are developed
The fusion of imaging means of image mode, can help us from different levels disease is appreciated and understood, and accomplish precisely to diagnose,
Reach personalized medicines.
With basic research deeply and the development of high throughput sequencing technologies, tumor formation mechanism to glioma and it is pernicious into
The mechanism study of exhibition obtains a series of important breakthroughs, while also determining the molecular marker of many keys relevant to glioma
Object (biomarkers), and corresponding biological agent is developed for these targets, but their therapeutic effect is unsatisfactory.
More and more researchs and viewpoint disclose: glioma is a kind of entity tumor that height is heterogeneous, is possessed a plurality of different carcinogenic
Access；In the different phase of Spatio-temporal Evolution, molecular marker is expressed, relied on oncogenic pathways also change correspondingly.Therefore, it is badly in need of
A kind of precisely diagnosis glioma is developed, accurately defines tumor boundaries molecular imaging probe and method, the diagnosis for glioma
It will be of great significance with treatment.
Summary of the invention
It is an object of the invention to provide a kind of double targetings of three mode for having nuclear-magnetism, optoacoustic, near-infrared fluorescent signal
Probe and its preparation method and application can significantly improve preoperative using the strategy of the exclusive bis- targetings of EGFR and SEC61G
Navigation application in the precision of diagnosing tumor and potential art, has broad application prospects and conversion value.
A kind of double multi-modal molecular image probes of targeting of the invention, are by epidermal growth factor EGF polypeptide and anti-SEC61G
Multi-modal molecular image probe of the antibody as targeting group targets neoplastic cells.
It further, is by EGF polypeptide and anti-SEC61G antibody coupling to the multimode for being able to achieve different kinds of molecules video imaging
On state signaling molecule group.
Further, the multi-modal signaling molecule group for being able to achieve different kinds of molecules video imaging includes: to have
Nuclear-magnetism, optoacoustic, near-infrared fluorescent signal three mode signals molecular radicals.
Further, it is described have nuclear-magnetism, optoacoustic, near-infrared fluorescent signal three mode signals molecular radicals include
The nano oxidized iron particle of superparamagnetic with MRIT2 enhancing contrasting effects of core, and there is near-infrared fluorescent and light
The indocyanine green of acoustic imaging signal.
The preparation method of double multi-modal molecular image probes of targeting of the present invention: synthesis lipophilic group package first
Superparamagnetic iron oxide nano inner core；It (include: again DSPE-PEG5000-COOH, DPSE- with DSPE-PEGization phosphatide
PEG5000-NHS, DSPE-PEG2000-COOH or DSPE-PEG2000-NHS, being purchased from Ai Weituo (Shanghai) medical sci-tech has
Limit company) package, so that it is switched to water solubility；Then indocyanine green is introduced in the hydrophobic region of product；Finally pass through NHS/EDC again
The reaction activated carboxyl of (1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride/n-hydroxysuccinimide), couples
Upper corresponding antibody and polypeptide simultaneously purify, that is, double targeted probes are made.
Further, the preparation method of double multi-modal molecular image probes of targeting of the present invention:
(1) lipophilicity oleyl amine/oleic acid chain package superparamagnetic iron oxide nano inner core is synthesized by high temperature thermal decomposition method, i.e.,
Hydrophobicity SPIO nano inner core；Rotary evaporation is recycled to carry out DSPE-PEGization phosphatide again to hydrophobicity SPIO nano inner core
Package makes it switch to water solubility；
(2) by the parents characteristic of indocyanine green (ICG), it is made independently to enter hydrophobicity under hydrophobic forces effect
The hydrophobic region of SPIO nano inner core and DSPE-PEGization phosphatide junction；
(3) the last reaction activated carboxyl for passing through NHS/EDC again, couples corresponding antibody or polypeptide and purifies, i.e.,
Double targeted probes are made.
Further, the preparation method of the multi-modal molecular image probe of double targetings, specifically comprises the following steps:
1) synthesis of hydrophobicity SPIO nano inner core: taking ferric acetyl acetonade 0.7g, oleic acid 4.45g, and oleyl amine 4.07g is placed in burning
It shakes and mixes in bottle, argon gas is passed through, with the air in exclusion system；120 DEG C are warming up in half an hour to react 2 hours；Then, exist
220 DEG C of reaction 30min are lifted temperature in half an hour；Finally the temperature was then adjusted to 300 DEG C of the reaction was continued 30min in 1 hour；To it
After natural cooling, centrifugation after addition 100ml ethyl alcohol mixes is resuspended after removing supernatant with 50ml hexane again so that product precipitating,
It so repeats to precipitate and be resuspended four periods, precipitating is finally collected by centrifugation, is dissolved in toluene, adjust its iron content as after 10mg/ml
It is placed in 4 DEG C of preservations；
2) it synthesizes DSPE-PEG5000-COOH coated water solubility SPIO: being contained with iron in hydrophobicity SPIO nano inner core
Amount and DSPE-PEG5000-COOH are placed in flask according to the ratio of mass ratio 1:5, and addition 20ml toluene is sufficiently vibrated molten
Solution；Open Rotary Evaporators, connect condensing unit (effect of condensation be so that by thermal evaporation toluene vapor restore liquid, into
Enter waste liquid bottle to be recycled, prevent from causing danger into air), revolving speed is adjusted to 40 revs/min, maintains temperature at 50 DEG C, to
After toluene evaporation is clean in system, 20ml deionized water is added, is vibrated 1 hour under ultrasound；By 0.22 μm of needle of system solution
Formula filter filter 23, removes big polymer；100000g ultracentrifugation is used again, is collected precipitating and is resuspended；(use 100000g
The unloaded micella of SPIO is not coated in ultracentrifugation removal system, because DSPE-PEG5000-COOH can be spontaneous in system
The empty carrier without wrapping up SPIO is formed, filtering can only remove big particle, but centrifugation can make these not wrap up
The empty carrier of SPIO is separated)；Precipitating is collected to be resuspended.
3) loading certainly of indocyanine green: according to quality than iron: indocyanine green is that indoles is added in 20:1 in water solubility SPIO
Cyanines are green, dialyse in vibrating 4h on shaking table, then with the bag filter of molecular weight 3500Da, first 4 hours respectively at 10 minutes, 30 points
Clock 60 minutes, 2 hours, carries out changing water for 4 hours, collects after staying overnight；
4) anti-SEC61G antibody (being purchased from Abcam, ab209867) and EGF polypeptide (being purchased from Abcam, ab9697) and nanometer
Carrier couples: in the nano-carrier system that step 3) obtains, taking SPIO concentration of iron is the solution 1ml, 5mgEDC of 1mg/ml
It is reacted at room temperature with 10mgNHS 15 minutes, carboxyl is activated, then gone with molecule interception for the super filter tube of 100Kd
Except the free EDC/NHS not reacted with activated carboxylic；Adjustment concentration of iron is 1mg/ml after precipitating is resuspended, and takes 1ml solution, is added
The EGF polypeptide solution of 10ul1mg/ml and the anti-SEC61G antibody-solutions of 10ul1mg/ml, after the reaction was continued at room temperature 4 hours,
Precipitating is collected by centrifugation, removes the protein molecular not coupled with carrier, is placed in 4 DEG C of refrigerators and saves backup.
The invention further relates to the applications of the multi-modal molecular image probe of double targetings, are used to prepare the system of diagnosing tumor
Agent is especially used to prepare the preparation of Diagnosing Gliomas.
Double multi-modal molecular image probes of targeting prepared by the present invention pass through dynamic light scattering (DLS), transmission electron microscope
(TEM), vibrating specimen magnetometer (VSM), ultraviolet-visible spectrophotometer (Uv-Vis) etc. survey its physicochemical property
It is fixed.
The in vitro toxicity detection of double multi-modal molecular image probes of targeting prepared by the present invention and tumour cell Targeting Performance
Research: the toxicity of cell is made using the nano-probe under CellCountingKit-8 (CCK-8) method detection various concentration gradient
With；In order to verify tumour cell in vitro to the intake of probe and targeting binding ability, by by cell and different targeting bases
The probe of group's label is incubated altogether, carries out qualitative and quantitative ratio to its Targeting Effect respectively using laser co-focusing and flow cytometry
Compared with.
Nuclear-magnetism-optoacoustic-fluorescence multi-modality imaging research of double multi-modal molecular image probes of targeting prepared by the present invention: logical
Cross tail vein injection mode will target with non-targeted nano-probe, by under each mode targeting group with non-targeted group at
As performance individually compares, the advantageous information for finally integrating three kinds of mode is analyzed comprehensively.
Advantage for present invention:
First, synthesis material of the present invention or component are all either ICG or SPIO, are all FDA approvals
Can be used for clinical material；
Second, maximum innovation of the invention is, find GBM patient's epidermal growth factor receptor (EGFR) and
There is the trend being overexpressed jointly in γ subunit (SEC61G) the two albumen of protein transporters Sec61.By targeting simultaneously
Two molecular targets can significantly improve sensitivity and the specificity of diagnosis.
Third can help to obtain clinician more more accurately in conjunction with the imaging characteristics of different image modes
Diagnostic message.Meanwhile fluorescence, optoacoustic and the NMR imaging effect of the permanent enhancing of molecular probe, it is preoperative diagnosis and operation rule
Draw and art in navigation information more abundant can be provided, make more reasonable resolution, with clinical application prospect and
Conversion value.
Detailed description of the invention
Fig. 1: Figure 1A nano-probe synthesis flow schematic diagram, Figure 1B are the transmission electron microscope pictures of SPIO, and Fig. 1 C is at room temperature
The hysteresis loop of SPIO, Fig. 1 D are the hydration grain size distributions of probe；Fig. 1 E is the different mixed spies of Iron:ICG mass ratio
Absorbance A of the needle in 400-900nm range of wavelength.
Fig. 2: TCGA (cancer gene group map) data analyze EGFR and SEC61G in the coexpression trend result of GBM；
A is the oncopoint and heatmap of EGFR and SEC61G in GBM patient,
B: in genome copy numbers level, the correlation of EGFR and SEC61G；
C: on mRNA expression, the correlation of EGFR and SEC61G；
Trend prediction analysis result coexists in D:EGFR and SEC61G；
The influence that its mRNA is expressed in the variation of E:SEC61G copy number；
The case of F:SEC61G amplification, prognosis are worse.
Fig. 3: Western Blot verifies the coexpression trend of EGFR and SEC61G in glioma cell line；
The protein quantification of EGFR and SEC61G in A:U87 and T98 cell；
B and C: gray value analyzes the coexpression trend of EGFR and SEC61G in U87 and T98.
Fig. 4: the toxicity detection of nano-probe.
Fig. 5: the qualitative and quantitative detection result of the external affinity of nano-probe and glioma cell line；
A: laser co-focusing laboratory qualitative is most strong with tumour cell affinity analysis shows that the more single targeting group of double targeting groups,
And the binding ability of this enhancing, it can be blocked by corresponding excessive free antibodies；
B: flow cytometry tests quantitative analysis shows the more single targeting group of double targeting groups, most strong with tumour cell affinity,
And the binding ability of this enhancing, it can be blocked by corresponding excessive free antibodies.
Fig. 6: the fluorescence imaging situation of the internal different time points of probe；
A: after double targeted probes tail vein injections 0,4,6,12,24,48h, the internal metabolic condition of probe and swollen
The fluorescence imaging at tumor position；
B: for 24 hours, the common location result of archebiosis the light imaging and fluorescence imaging of tumour；
C: the variation of the background area fluorescence intensity of the tumor region and opposite side of different time points；
D: the size of the imaging signal to noise ratio of different time point tumor regions.
Fig. 7: the T2 of probe different time points in vivo enhances contrasting effects；
The image (original image and pcolor) and tumor region of A and B: double target probe groups in situ tumor nuclear-magnetism T2 enhancings and
The variation of the T2 value of opposite side normal cerebral tissue；
C and D: non-target probe groups in situ tumor nuclear-magnetism T2 enhancing image (original image and pcolor) and tumor region and
The variation of the T2 value of opposite side normal cerebral tissue.
Fig. 8: the photoacoustic imaging and histology verification result of probe in vivo；
A: double targeting groups are dyed with non-targeted group of probe in the imaging contrast on tumor region and boundary and corresponding H&E；
B: double targeting groups are compared with non-targeted group of probe is in the signal strength on tumor region and boundary；
C: the immunohistochemical staining result of tumor tissues EGFR and SEC61G entirety；
D: the partial enlargement picture of tumor tissues EGFR and SEC61G immunohistochemical staining.
Specific embodiment
Further describe the present invention below with reference to specific example, the features of the present invention also can with the description of example and more
It is clear.But these examples are only exemplary, do not constitute any restrictions to protection scope of the present invention.
Embodiment one: probe preparation process
1. the synthesis of hydrophobicity SPIO nano inner core: taking ferric acetyl acetonade 0.7g, oleic acid 4.45g, oleyl amine 4.07g is placed in
It shakes and mixes in 100ml twoport flask, be fixed on constant temperature blender with magnetic force, access conduit is passed through argon gas 5min, to squeeze system
Interior air.Power on and heated to system, after system is warming up to 120 DEG C within half an hour, temperature is maintained to react at 120 DEG C
2 hours；Then, 220 DEG C are lifted temperature to gradually within half an hour, and system is controlled in 220 DEG C of reaction 30min；Finally at 1 hour
It is interior the temperature was then adjusted to after 300 DEG C, in the reaction was continued with this condition 30min.After its natural cooling, by four periods
Ethyl alcohol and hexane are precipitated and are resuspended respectively, and precipitating is finally collected by centrifugation, is dissolved in toluene, and adjusting its iron content is 10mg/ml
It is placed on 4 DEG C of long-term preservations.
2. synthesizing the coated water solubility SPIO of DSPE-PEG-COOH-5000: with the content and DSPE-PEG- of iron in SPIO
COOH-5000 is placed in flask according to the ratio of mass ratio 1:5, and 20ml toluene is added and carries out sufficiently oscillation dissolution；Open condensation
Device and vacuum pump adjust revolving speed to 40 revs/min, maintain temperature at 50 DEG C, after toluene evaporation in system is clean, are added
20ml deionized water vibrates 1 hour under ultrasound；By system solution 0.22um syringe filter filter 23, big gather is removed
Close object；Again with the unloaded micella for not being coated with SPIO in 100000g ultracentrifugation removal system；Precipitating is collected to be resuspended.
3.ICG's loads certainly: according to the iron in mass ratio SPIO: ICG is respectively 5:1, the past water solubility SPIO of 10:1,20:1
Interior addition ICG dialyses in vibrating 4h on shaking table, then with the bag filter of molecular weight 3500Da, and first 4 hours respectively at 10 points
Clock 30 minutes, 60 minutes, 2 hours, carries out changing water for 4 hours, collects after staying overnight.
4. step 3 obtain nano-carrier system in, take SPIO concentration of iron be 1mg/ml solution 1ml, 5mgEDC and
10mgNHS reacts 15 minutes at room temperature, activates to carboxyl, is then removed with molecule interception for the super filter tube of 100Kd
The free EDC/NHS not reacted with activated carboxylic；Adjustment concentration of iron is 1mg/ml after precipitating is resuspended, and takes 1ml solution, is added
The EGF polypeptide solution of 10ul1mg/ml and the anti-SEC61G antibody-solutions of 10ul1mg/ml, after the reaction was continued at room temperature 4 hours,
Precipitating is collected by centrifugation, removes the protein molecular not coupled with carrier, is placed in 4 DEG C of refrigerators and saves backup.
5. vitro characterization
Transmission electron microscopy: it takes appropriate drop in copper mesh, is scanned after natural drying with transmission electron microscope.Electronic Speculum operates in
Physics and chemistry research institute, institute, section instructs personnel to help through；After the completion of scanning, its particle size is measured, its whole pattern is observed, sees
Figure 1B.It can be seen that SPIO Nanoparticle shape is relatively regular, in almost spherical either ellipse；It is uniformly dispersed, it is not bright
Aobvious aggregation illustrates that its stability is preferable.Available by measuring: the diameter of SPIO kernel is in 12.26 ± 0.35nm or so.
It is saturated the measurement of magnetic intensity: utilizing magnetic vibration sample magnetometer (Vibration sample
Magnetometer, VSM), at room temperature, the sample 1mg of drying is weighed, is put into specimen cup and is pressed with non-magnetic plastic package
It is real, then start to measure, record and save experimental data, draws hysteresis loop, judge its superparamagnetism and Relaxivity.By
Hysteresis loop result can be seen that (Fig. 1 C): it is about 65emu/g that it, which is saturated magnetic intensity, and (curve is by former with superparamagnetism
Point), it can be used as the contrast agent of suitable MRIT2 enhancing scanning.
Be hydrated the measurement of partial size: solution to be measured uses sonic oscillation 10min before testing, and cuvette is cleaned with deionized water
It is rinsed later with alcohol, hair dryer Quick-air-drying；Then 1ul solution to be measured is added to the colorimetric that 1ml deionized water is housed
It in ware, is put into Malvern particle instrument and is detected after mixing well, vary the cleaning step of stressed front every time.It is measured
Experimental data is saved and exported afterwards, draws, sees Fig. 1 D.It can be seen that the nanometer for having coupled EGF and anti-SEC61G antibody is visited
Needle average grain diameter is 43.80nm or so.
The measurement of absorbance: with before measurement of ultraviolet-visible spectrophotometer absorbance, take two quartz colorimetric utensils go from
Sub- water is cleaned, alcohol rinsing carries out flash baking；Most start to make baseline calibration: all adding 3ml deionized water in two cuvettes
It is calibrated, sets wave-length coverage between 400-900nm.The deionized water of one of cuvette is changed into together after completion
The testing liquid of volume measures its absorbance in this wave-length coverage by several times respectively.Record and preservation experimental data, are shown in figure
1E: as the increase of ICG specific gravity can have apparent absorption peak in 780nm or so, and the value of absorbance is consequently increased,
The amount for showing that ICG is loaded is consequently increased.On the contrary, in the case where only SPIO, be not found have in this wave-length coverage it is bright
The appearance of aobvious absorption peak.
Embodiment two: double targeted probes in GBM EGFR and SEC61G detect
1, as can be seen from the result of figure 2 that: EGFR and SEC61G is whether horizontal or mRNA's in genome copy numbers
On expression pattern, there is the trend of " symbiosis "；In the research queue that we select, 33% GBM patient is in genome water
There is the amplification of SEC61G gene copy number on flat；It is interesting that these GBM patients with SEC61G amplification event, while
All has the amplification of EGFR gene；It has a direct impact: having moreover, this amplification event can be formed the transcription of its mRNA
The case of amplification event, the level for corresponding to mRNA also dramatically increase (Fig. 2A)；Meanwhile EGFR and SEC61G are in genomic level
On the variation of copy number tend to synchronous (Fig. 2 B)；And in the waterborne of mRNA, the Spearman of the expression of two albumen
Related coefficient is 0.58, and Pearson correlation coefficients 0.65, P value is respectively less than 0.05, is had statistical significance (Fig. 2 C).Thus may be used
With determination, this trend coexisted of EGFR and SEC61G are not haphazard part.Furthermore SEC61G is copied on genome
The transcription of several increases, mRNA is consequently increased, and difference has statistical significance (Fig. 2 E).Finally, as EGFR,
SEC61G similarly can be as the index of the Index for diagnosis of GBM patient, i.e., when SEC61G is highly expressed, patient's
Prognosis is worse (P < 0.05, Fig. 2 F).
2. western blot Western Blot: to cell climbing sheet in culture dish to 80% or so, discarding culture medium, PBS is clear
200ul cell pyrolysis liquid is added after washing 3 times, is placed in 2 minutes on ice；Adherent part is then scraped with cell scraper.Collection is split
After solution liquid is centrifuged, supernatant is taken to carry out protein quantification with BCA method；Loading is carried out after taking same volume sample to boil again, electrophoresis, is turned
Film and closing；Primary antibody EGFR and SEC61G dilution ratio is 1:5000, selects GAPDH as internal reference albumen, incubates in 4 DEG C of refrigerators
Secondary antibody incubation is developed for 1 hour again after educating；Gray value record and analysis finally are carried out to image band value.As a result see Fig. 3,
It can be seen that the expression of EGFR is relatively low, SEC61G's in U87 cell (source of people human malignant glioma cell line U87)
Expression is relatively low；And in T98 cell line (human glioma cell line), the expression of EGFR increases, then SEC61G
Expression also increase.
3. the toxicity detection of probe: after the cell of adhere-wall culture is carried out digestion collection, adjusting its number of cells is 106
A/ml, then with 104The cell concentration in a/hole is layered in 96 orifice plates, is placed in constant incubator and is grown overnight.By SPIO,
5 kinds of SPIO-ICG, SPIO-ICG-EGF, SPIO-ICG-SEC61G, SPIO-ICG-Dual (EGF and SEC61G) etc. not isolabeling
Molecular probe with iron content for 0,5,10,20,40,60,80ug/ml gradient and cell be incubated for altogether 48 hours.Upper machine testing
Liquid is changed in cleaning in first 2 hours, and the CCK-8 of every hole addition 10ul (100ul mixed liquor) is cultivated, at room temperature every half an hour enzyme mark
Instrument measures its light absorption value (OD value) after dosing in 4 hours at 450nm, calculates cell viability according to following formula to assess
The toxicity size of probe:
Cell viability (%)=[A dosing-A blank]/[A0 dosing-A blank] X100%
Wherein:
A blank: there is culture medium, the absorbance in CCK-8 and cell-free each hole
A0 dosing: the absorbance in each hole without nano-probe with culture medium, cell and CCK-8
A dosing: the absorbance in each hole with culture medium, cell, nano-probe and CCK-8.
As a result see Fig. 4: no matter whether nano-probe has coupled targeting group, under different concentration with cell incubation 48h,
With the increase of concentration and probe concentration, the effect that cell growth is suppressed is become apparent from, but even if is up at concentration (Iron)
When 80ug/ml, the activity of cell entirety remains within 85% or more, shows it in vitro to the toxicity of cell and growth
The effect of inhibition is not significant.Therefore, it is suitable as contrast agent and is used for In vivo detection.
4. laser co-focusing experiment and flow cytometry carry out qualitative and quantitative inspection to the external affinity of double target image probes
It surveys
Laser co-focusing experiment: 10 are deployed into after taking eugonic cell, digestion to be collected by centrifugation6The cell of a/ml is outstanding
Liquid.Take 105A cell is added in laser co-focusing capsule and cultivates for 24 hours, and PBS is cleaned 3 times, is added after probe is incubated for 2h and uses poly first
30min is fixed in aldehyde, and Qula is added the DAPI dyeing anti-fluorescence quencher mounting of 3min after leading to rupture of membranes 5min, carries out machine examination
It surveys.
Flow cytometry: taking cell count concentration is 106The cell suspension of a/ml, with 1056 holes are added in the concentration in a/hole
Plate is incubated overnight, and PBS is added probe and is incubated for 2h after cleaning 3 times, and cell shape precipitating is collected after centrifugation in digestion, and PBS is carried out after being resuspended
Upper machine testing.
Note:
A. probe used in laser co-focusing and flow cytometry has passed through NHS-FITC to antibody or polypeptide point
Son carries out fluorescent marker.The fluorescent collecting in the channel FITC can directly be carried out.
B. blocking experiment is incubated for probe altogether again after being pre-processed using free 20 times excessive free antibodies or polypeptide.
By the result of laser co-focusing we can see that (Fig. 5 A): the bis- target image nano-probes of EGFR and SEC61G compared to
For the mono- targeting of EGFR and the mono- targeting group of SEC61G, is more absorbed or combined by tumour cell；Moreover, this binding ability
Enhancing can be by carrying out part blocks using corresponding excessive free antibodies or polypeptide in advance.It can be seen that: there is target
To group (EGF polypeptide or anti-SEC61G antibody) as guidance under the premise of, while having double targets of EGFR and SEC61G can be with
More it is obviously improved intake and binding ability of the tumour cell to nano-probe.
Meanwhile finding (Fig. 5 B) after carrying out flow cytometer detection: the ratio of single EGF targeting group probe combination cell is 37.5%,
Single SEC61G targeting group probe cell combination ratio is 48.2%, and EGFR and SEC61G group probe can reach highest combination ratio
Rate 63.8%；There is targeting group (EGF and SEC61G) to verify the binding ability of this enhancing.Equally, we
Processing is incubated in advance with corresponding excess, free antibodies or polypeptide progress is taken before cell is incubated for altogether in every group of probe respectively, is tied
Fruit, which shows, can different degrees of blocking effect: the blocking group of blocking group and SEC61G for EGFR has 10% He respectively
The effect that 23.6% blocking combines；Moreover, mutually being echoed with laser co-focusing result, the resistance to the bis- targeting groups of EGFR and SEC61G
Disconnected effect be also it is most significant, up to 32.6%.
Embodiment three: double target image probes are in the fluorescence of body, nuclear-magnetism and photoacoustic imaging effect experiment
1. the foundation of animal model
The building of subcutaneous tumor model: selection in Exponential growth stage, cell that cell viability is strong digested, from
The heart, collection, PBS clean 3 times after by PBS: matrigel according to volume ratio be 1:1 in a manner of uniformly mixed, and adjust cell
Concentration is to 107A/ml；100ul mixing with cells suspension is drawn at nude mice back to be subcutaneously injected with insulin needle, works as needle point
Parallel inserting needle 2-3mm again when having breakthrough to feel, is careful not to puncture skin, slowly inject, and cutaneous protuberance whiting indicates to be injected into
Mouse is put back to and continues to raise by function, routine observation, when its diameter about 0.5mm can be tested.
The building of heteroplastic transplantation brain tumor model in situ: good thin of growth vigorous (be in increased logarithmic phase), vigor is chosen
Born of the same parents carry out PBS cleaning, pancreatin digestion after be collected by centrifugation, exhaust surplus liquid, with PBS: matrigel 1:1 mix in equal volume after to thin
Born of the same parents are resuspended, and adjustment concentration is 107A/ml is placed in ice chest or spare on ice.It is three-dimensional fixed to be fixed on after mouse anesthesia
On the instrument microscope carrier of position, carried out disinfection 3 times using Iodophor to its head；Scalp, the visual field are carefully cut along middle line using sterile scissors
It is advisable with clear exposure hat, sagittal suture and median line.Entry point is selected in right side brain, and apart from sagittal suture 1.5mm, coronal suture is past
2mm afterwards.First is penetrated for skull with osteoclastic but not enter brain parenchym；It beaten, mixed with finger tip again again before per injection
Even cell suspension then slowly draws 6ul cell suspension (avoiding generating bubble) with micro liquid inlet device, is fixed and finds elder generation
Preceding osteoclastic needle entry point vertically slowly enters brain parenchym, first inserting needle 3mm, then the 0.5mm that shrinks back, can inject cell suspension, push away
Whether there is cell suspension to overflow (if any wiping is removed in time, and adjusts the depth of inserting needle) at observation inserting needle during note；Often push away
Into 1ul cell suspension, interval 30s until whole liquid inject completion, when injecting completion for the last time, keeps needle repeatedly
Situations such as slowly pulling out needle again after stopping 2 minutes, paying attention to the breathing of observation mouse in the process.Then scalp is sutured,
Disinfection.It puts back in cage to its revival.The growth detection of intracranial tumors is monitored by IVIS or MRI, to tumour growth to conjunction
Suitable size, for carrying out subsequent experimental.
2. multi-modality imaging in body
Small animal living body fluorescence and archebiosis light detection: probe is administered by the mode of tail vein injection, gas fiber crops
After liquor-saturated, 0h, 1h, 2h, 4h, 6h, 12h upon administration, for 24 hours, 48h with small animal living body imager to the fluorescence signal of probe into
Row detection, according to the close wave band of excitation-emission of ICG, select 780nm and 810nm respectively as excitation and launch wavelength into
Row signal acquisition；Acquisition for archebiosis light, every mouse extract the D- fluorescein 100ul of 15mg/ml through being injected intraperitoneally,
Switching signal acquisition module is BLI acquisition module, and 10-15min carries out the acquisition of image after substrate injection.As a result see Fig. 6: from
Tail vein injection probe starts, the time point before 6 hours, we be difficult directly to distinguish on living body tumour and around tissue
Region, fluorescence signal are concentrated mainly on liver area；In 12 hours this time points, the comparison of tumour and surrounding tissue was gradually bright
Aobvious, when reaching for 24 hours, signal increases to maximum in the aggregation of tumor region, and tumour at this time: the signal-to-noise ratio of background reaches highest
(1.974 ± 0.061), and it is this it is apparent comparison until 48h be also maintained at high levels (1.830 ± 0.027) (figure A, C,
D).In order to be verified directly under condition of living organism where enhancing contrast district is tumour really under fluorescence imaging, we for 24 hours this
The luciferase substrate D- fluorescein of 15mg/kg is injected intraperitoneally in time point, since tumor cells expression has luciferase, thus by
With the effect of D- fluorescein, the higher archebiosis light of specificity is formed.By (Fig. 6 B) it can be seen that excitation fluorescence area and life
In the fusion of tumor locus, the region so as to confirm that fluorescence highlights under condition of living organism is exactly tumour institute in object self-luminous region
?.
The acquisition of toy nuclear magnetic data: after experiment mice is grouped, being administered by tail vein, obtain 0h, 4h, 12h respectively,
For 24 hours with the nuclear-magnetism image of the tumor region of 48h different time points, basic parameter is set as；FOV 40mm, Scan slice thickness
0.8mm, TR=6000ms, TE=60ms.Then by the T2 value of measurement tumor region, it is compared.In order to preferably intuitive
Show this variation tendency (because SPIO is negativity contrasting effects), we depict corresponding pcolor and show, as a result
See Fig. 7: enhancing the visualisation range of decrease in 24 hours T2 with the tumor region of the bis- targeted probes imaging groups of EGFR and SEC61G
Up to 34.3% (T2 average value drops to 12458.8ms from 18968.4ms), variation of this variation compared with opposite side normal cerebral tissue
Significantly, the reduction of signal value is more obvious (Fig. 7 A and B)；And the mouse of non-targeted probe imaging is used, the T2 of tumor region
The reinforcing effect range of decrease is only 10.9% (average from 18020.8ms to 16060.5ms).From the color change width of corresponding pcolor
Degree can more intuitive this variation tendency (Fig. 7 C and D) of display；.
The acquisition of toy photoacoustic data: experiment mice is fixed on load after the administration of tail vein approach under narcosis
On platform, in order to reduce artifact, ultrasonic coupling liquid is smeared to imaging region.Before measurement, the signal (ICG) that selection needs to be imaged is inhaled
Receive spectrum, setting scanning wavelength is while to choose Hb and HbO for 720nm, 740nm, 780nm, 810nm, 830nm2Absorption light
Spectrum, according to the timing information of chemiluminescence assay, it is also considered that it is bigger to load of the photoacoustic imaging to toy, select several spies
Other time point (0h, for 24 hours, 48h) signal acquisition is carried out, as a result see Fig. 8.It can be seen that the bis- targeting labels of EGFR and SEC61G
The outer region for the display tumour that nano-probe can be more clear is not significant for the enhancing in the centre of tumour；And
For non-targeted group, it is not only very difficult for the judgement of tumor region and profile on the whole, but also does not have double
The apparent edge strengthening of targeting group (Fig. 8 A and B).We carry out EGFR respectively after being sliced to tumor tissues and SEC61G exempts from
The discovery of epidemic disease histochemical staining: EGFR and SEC61G is stronger in the expression of the periphery of tumour, and closer to central area, express weaker (figure
8C and D).
Claims (9)
1. a kind of double multi-modal molecular image probes of targeting, it is characterised in that: be by epidermal growth factor EGF polypeptide and to resist
Multi-modal molecular image probe of the SEC61G antibody as targeting group targets neoplastic cells.
2. double multi-modal molecular image probes of targeting according to claim 1, it is characterised in that: be by EGF polypeptide and to resist
On SEC61G antibody coupling to the multi-modal signaling molecule group for being able to achieve different kinds of molecules video imaging.
3. double multi-modal molecular image probes of targeting according to claim 2, it is characterised in that: described being able to achieve is a variety of
The multi-modal signaling molecule group of molecular image imaging includes: the three mode letter for having nuclear-magnetism, optoacoustic, near-infrared fluorescent signal
Number molecular radical.
4. double multi-modal molecular image probes of targeting according to claim 3, it is characterised in that: described has core
Magnetic, optoacoustic, near-infrared fluorescent signal three mode signals molecular radicals include core have MRIT2 enhance contrasting effects
The nano oxidized iron particle of superparamagnetic, and the indocyanine green with near-infrared fluorescent and photoacoustic imaging signal.
5. the preparation method of the described in any item double multi-modal molecular image probes of targeting of claim 1-4, it is characterised in that: first
The first superparamagnetic iron oxide nano inner core of synthesis lipophilic group package；Again with DSPE-PEGization phosphatide package, it is made to switch to water
Dissolubility；Then indocyanine green is introduced in the hydrophobic region of product；The reaction activated carboxyl for finally passing through EDC/NHS again, couples
Double targeted probes are made in corresponding antibody and polypeptide.
6. the preparation method of double multi-modal molecular image probes of targeting according to claim 5, it is characterised in that:
(1) lipophilicity oleyl amine/oleic acid chain package superparamagnetic iron oxide nano inner core is synthesized by high temperature thermal decomposition method, i.e., it is hydrophobic
Property SPIO nano inner core；Rotary evaporation is recycled to be wrapped up DSPE-PEGization phosphatide hydrophobicity SPIO nano inner core again
It is set to switch to water solubility；
(2) by the parents characteristic of indocyanine green, enter it independently in SPIO nanometers of hydrophobicity under hydrophobic forces effect
The hydrophobic region of core and DSPE-PEGization phosphatide junction；
(3) the last reaction activated carboxyl for passing through NHS/EDC again, couples corresponding antibody or polypeptide and purifies, that is, be made
Double targeted probes.
7. the preparation method of double multi-modal molecular image probes of targeting according to claim 5, it is characterised in that:
Specifically comprise the following steps:
1) synthesis of hydrophobicity SPIO nano inner core: taking ferric acetyl acetonade 0.7g, oleic acid 4.45g, and oleyl amine 4.07g is placed in flask
Concussion mixes, and argon gas is passed through, to drain the air in system；120 DEG C are warming up in half an hour to react 2 hours；Then, small half
When interior lift temperature to 220 DEG C of reaction 30min；Finally the temperature was then adjusted to 300 DEG C of the reaction was continued 30min in 1 hour；To its nature
After cooling, it is centrifuged after the mixing of 100ml ethyl alcohol is added so that product precipitating, is resuspended after removing supernatant with 50ml hexane, so again
It repeats four precipitatings and the period is resuspended, precipitating is finally collected by centrifugation, is dissolved in toluene, adjusting its iron content is that 10mg/ml is placed on 4 DEG C
It saves；
2) synthesize the coated water solubility SPIO of DSPE-PEG5000-COOH: with the content of iron in hydrophobicity SPIO nano inner core and
DSPE-PEG5000-COOH is placed in flask according to the ratio of mass ratio 1:5, and 20ml toluene is added and carries out sufficiently oscillation dissolution；
Rotary Evaporators are opened, condensing unit is connected, adjust revolving speed to 40 revs/min, reaction temperature is set at 50 DEG C, to first in system
After benzene evaporation is clean, 20ml deionized water is added, is vibrated 1 hour under ultrasound；By 0.22 μm of syringe filter of system solution
Filter 23, removes big polymer；100000g ultracentrifugation is used again, is collected precipitating and is resuspended；
3) loading certainly of indocyanine green: according to quality than iron: indocyanine green is that indocyanine green is added in 20:1 in water solubility SPIO,
It dialyses in vibrating 4h on shaking table, then with the bag filter of molecular weight 3500Da, first 4 hours respectively at 10 minutes, 30 minutes, 60
Minute, it 2 hours, carries out within 4 hours changing water, be collected after staying overnight；
4) anti-SEC61G antibody and EGF polypeptide and nano-carrier couple: in the nano-carrier system that step 3) obtains, taking
SPIO concentration of iron is that the solution 1ml, 5mgEDC and 10mgNHS of 1mg/ml reacts 15 minutes at room temperature, is activated to carboxyl,
The free EDC/NHS not reacted with activated carboxylic is then removed for the super filter tube of 100Kd with molecule interception；After precipitating is resuspended
Adjustment concentration of iron is 1mg/ml, takes 1ml solution, and the EGF polypeptide solution of 10ul1mg/ml and resisting for 10ul1mg/ml is added
SEC61G antibody-solutions after the reaction was continued at room temperature 4 hours, are collected by centrifugation precipitating, remove the protein molecular not coupled with carrier,
4 DEG C of refrigerators are placed in save backup.
8. any one of the multi-modal molecular image probe of described in any item pairs of targetings of claim 1-4 or claim 5-7 institute
The application for the multi-modal molecular image probe of double targetings that the preparation method stated obtains, which is characterized in that be used to prepare diagnosing tumor
Preparation.
9. the application of double multi-modal molecular image probes of targeting according to claim 8, which is characterized in that be used to prepare brain
The preparation of diagnosis of glioma.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112546224A (en) * | 2020-12-22 | 2021-03-26 | 暨南大学 | Preparation method of bimodal non-small cell lung cancer targeted nanoprobe based on ultra-small magnetic nanoparticles |
| CN112843258A (en) * | 2021-01-28 | 2021-05-28 | 厦门大学附属翔安医院 | Breast cancer targeted molecular probe and preparation method and application thereof |
| CN114107435A (en) * | 2021-11-30 | 2022-03-01 | 广东省人民医院 | Activatable photoacoustic-fluorescence dual-mode probe for real-time monitoring of immunotherapy and application |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102573925A (en) * | 2009-10-05 | 2012-07-11 | 佳能株式会社 | Contrast agent for photoacoustic imaging and photoacoustic imaging method utilizing same |
| CN105477630A (en) * | 2015-11-17 | 2016-04-13 | 华中科技大学 | Method capable of promoting migration of dendritic cells to lymph nodes and achieving multi-mode imaging simultaneously |
| CN105833296A (en) * | 2016-05-16 | 2016-08-10 | 东南大学 | Dual-mode optical imaging probe with tumor double-targeting function and preparation and application thereof |
| CN106692993A (en) * | 2017-01-04 | 2017-05-24 | 中国科学院自动化研究所 | Specific targeted magnetic resonance-optical dual-mode imaging probe and preparation method thereof |
| CN107050452A (en) * | 2017-05-12 | 2017-08-18 | 东南大学 | A kind of aptamer modified magnetic nanoparticle and its preparation method and application |
| CN109125743A (en) * | 2018-10-26 | 2019-01-04 | 复旦大学附属华山医院 | For Gliblastoma targeted nano mr contrast agent and preparation, application |
| CN109395101A (en) * | 2018-09-27 | 2019-03-01 | 复旦大学附属华山医院 | Target the preparation method of the mr contrast agent of blood-brain barrier and glioma |
-
2019
- 2019-06-20 CN CN201910537283.6A patent/CN110354281B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102573925A (en) * | 2009-10-05 | 2012-07-11 | 佳能株式会社 | Contrast agent for photoacoustic imaging and photoacoustic imaging method utilizing same |
| CN105477630A (en) * | 2015-11-17 | 2016-04-13 | 华中科技大学 | Method capable of promoting migration of dendritic cells to lymph nodes and achieving multi-mode imaging simultaneously |
| CN105833296A (en) * | 2016-05-16 | 2016-08-10 | 东南大学 | Dual-mode optical imaging probe with tumor double-targeting function and preparation and application thereof |
| CN106692993A (en) * | 2017-01-04 | 2017-05-24 | 中国科学院自动化研究所 | Specific targeted magnetic resonance-optical dual-mode imaging probe and preparation method thereof |
| CN107050452A (en) * | 2017-05-12 | 2017-08-18 | 东南大学 | A kind of aptamer modified magnetic nanoparticle and its preparation method and application |
| CN109395101A (en) * | 2018-09-27 | 2019-03-01 | 复旦大学附属华山医院 | Target the preparation method of the mr contrast agent of blood-brain barrier and glioma |
| CN109125743A (en) * | 2018-10-26 | 2019-01-04 | 复旦大学附属华山医院 | For Gliblastoma targeted nano mr contrast agent and preparation, application |
Non-Patent Citations (2)
| Title |
|---|
| BO LIU等: "Identification of SEC61G as a Novel Prognostic Marker for Predicting Survival and Response to Therapies in Patients with Glioblastoma", 《MED SCI MONIT》 * |
| CHUANG GAO等: "Near-infrared dye-loaded magnetic nanoparticles as photoacoustic contrast agent for enhanced tumor imaging", 《CANCER BIOL MED》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112546224A (en) * | 2020-12-22 | 2021-03-26 | 暨南大学 | Preparation method of bimodal non-small cell lung cancer targeted nanoprobe based on ultra-small magnetic nanoparticles |
| CN112843258A (en) * | 2021-01-28 | 2021-05-28 | 厦门大学附属翔安医院 | Breast cancer targeted molecular probe and preparation method and application thereof |
| CN114107435A (en) * | 2021-11-30 | 2022-03-01 | 广东省人民医院 | Activatable photoacoustic-fluorescence dual-mode probe for real-time monitoring of immunotherapy and application |
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