CN106540267A - Adr GNs coupled complexes for targeted therapy of malignant and preparation method thereof - Google Patents
Adr GNs coupled complexes for targeted therapy of malignant and preparation method thereof Download PDFInfo
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Abstract
Adr GNs coupled complexes for targeted therapy of malignant and preparation method thereof, belong to biomedicine technical field, gold nanoshell solution and the mixing of mPEG SH 2000 are reacted, the gold nanoshell of PEG modifications is obtained；The gold nanoshell solution mixing that again Adr and PEG is modified is reacted, and Adr GNs coupled complexes are obtained.The purpose that Adr is coupled on the GNs of PEG modifications is constituted with this.Present invention process has the advantages that few, with short production cycle operating procedure, low production cost, operating condition are gentle, coupling efficiency is high, by-product is few, pollution-free, product separation purification is simple.Adr GNs coupled complexes and the tumor cell to In vitro culture that is used in combination of siRap2b GNs coupled complexes prepared by the present invention shows obvious lethal effect.
Description
Technical field
The invention belongs to biomedicine technical field, it is intended to prepare a kind of new formulation, for oncotherapy, classification number is
A61P。
Background technology
Cancer, i.e. malignant tumor, are diseases that a class seriously threatens human health.According to IARC（IARC）Send out
The latest data of cloth shows that the whole world has more than 800 ten thousand people to die from cancer every year, and to the year two thousand twenty whole world, newly-increased cancer patient will surpass every year
17,000,000 are crossed, 3,000,000 by 2012 are risen to nearly 4,000,000 by China's cancer year new cases, therefore develop efficient anti-swollen
Tumor preparation, the significantly more efficient cancer immunotherapies of formulation are one of current urgent problems.
The formation of malignant tumor is relevant with many factors, and the exception of wherein caused by tumor suppressor p 53 and its signal path exists
Very important effect is played in the generation evolution of tumor.It is reported that, about 50% human tumor has p53 genes
Mutation, and in the tumor that p53 is not mutated, the approach relevant with p53 activation is all destroyed mostly, so as to cause p53 to lose
It is living.Therefore, for many years, p53 is always the emphasis of tumor research, and p53 signal path molecules are also the important target of oncotherapy
One of point.
Research finds that the new target gene Rap2b of p53 has the function of promoting tumor cell existence, reduces the intracellular of Rap2b
Expression can significantly improve the effect of amycin killing tumor cell, it is possible thereby to it is right to think that Rap2b is produced with tumor cell
The drug resistance of amycin is relevant.Further study show that, Rap2b is highly conserved in evolution, and in about 80% tumor tissues be in
Existing high expression.Therefore, Rap2b can be used for the treatment of tumor as a kind of important target spot in theory, by small molecules interference RNA
（siRNA）The intracellular expression level for reducing Rap2b can significantly improve fragmentation effect of the amycin to tumor cell.
Targeting preparation is the focus of current medical oncology research, and antitumor drug can be transported to target to greatest extent for it
Organ, reduces medicine distribution in the normal tissue, as far as possible so as to reach the therapeutic effect of high-efficiency low-toxicity.In recent years, based on receiving
The targeting drug delivery system of rice technology is widely used in the diagnosis and treatment of cancer.Particle diameter 10-1000 nm Nano medication due to small size
Effect and there is huge surface energy, be consequently belonging to thermodynamic unstable system and dynamic stabilization system.This has them
The properties such as many excellent magnetics for being different from body phase material, photoelectricity, photo-thermal.Additionally, in neoplasm targeted therapy application, these
The most important speciality of nanoparticle is exactly the high-permeability to solid tumor and retention effect --- EPR effects, i.e., relative to normal
Tissue, nanoparticle have tend to be gathered in the property of tumor tissues.Microvascular endothelial gap in normal structure is fine and close,
Structural integrity, macromole and lipid granule and nanoparticle are difficult to pass through blood vessel wall, and solid tumor mass medium vessels is abundant, blood
Tube wall gap is wider, poor structural integrity, lymphatic return disappearance, causes the selective high-permeability of nanoparticle and delay
Property.This phenomenon enables the medicine targeting drug release loaded by nanoparticle to tumor tissues and thin in Efficient killing effect tumor
While born of the same parents, the toxicity to normal body tissue is reduced.
The Major Difficulties that gene therapy is carried out using siRNA under environment in vivo are the RNase pair for existing in a large number in vivo
The degraded of exogenous RNA interference sequence, therefore exploitation can suppress the suitable pharmaceutical carriers of RNA degradeds for improving siRNA in vivo
Stability in environment is significant so as to improve curative effect.Current fast-developing nano material technology is then to solve this
Problem provides an important thinking.In various nano materials, nanometer gold is one of conventional gene drug carriers.
The content of the invention
Present invention aim at providing the Adr-GNs coupled complexes for targeted therapy of malignant.
The Adr-GNs coupled complexes for being used for targeted therapy of malignant in the present invention are characterized in that：In the gold of PEG modifications
Amycin is coupled in nanoshell（Adr）.
Research shows that golden nanometer particle can be with power supplying groups（Such as sulfydryl, amino etc.）Form stable chemical bond.This
Characteristic ensure that golden nanometer particle connects multi-medicament easily by chemical modification.For example, gene class medicine（Such as siRNA etc.）With
Higher density is modified in golden nanometer particle surrounding by mercapto gold key, space bit caused by the space conformation of nanoparticle surface
Resistance defines a kind of microenvironment along with the electron cloud of high density nucleic acid shell, during this microenvironment can suppress blood circulation
Degradation of the RNase to siRNA, so as to ensure that stability of the siRNA in vivo under environment.
The present invention is coupled Adr, for the targeted therapy of malignant tumor from hollow gold nanoshell as pharmaceutical carrier.This
Invention has all advantages of golden nanometer particle using GNs with Rap2b as target, can be by medicine（Adr）Stability and high efficiency ground is fixed
To tumor tissues are delivered to, the Efficient killing effect to tumor cell is realized, so as to substantially increase oncotherapy effect.
Present invention also offers the preparation method of above coupled complex.
Preparation method for the Adr-GNs coupled complexes of targeted therapy of malignant is comprised the following steps：
1）Gold nanoshell solution and mPEG-SH-2000 mixing are reacted, the gold nanoshell of PEG modifications is obtained；
2）The gold nanoshell solution mixing that Adr and PEG is modified is reacted, and Adr-GNs coupled complexes are obtained.
The present invention is in step 1）It is middle that GNs is modified using PEG first, the stability of nanoparticle can be improved.In step 2）In
The GNs solution that Adr is modified with PEG mixes and is reacted, and makes 5 ' end sulfydryls of the active amino of Adr form coordinate bond with GNs.
The purpose that Adr is coupled on the GNs of PEG modifications is constituted with this.
Present invention process has that few, with short production cycle operating procedure, low production cost, operating condition be gentle, coupling efficiency
It is high, the advantages of by-product is few, pollution-free, product separation purification is simple.
Adr-GNs coupled complexes and siRap2b-GNs coupled complexes prepared by the present invention is used in combination to external
The tumor cell of culture shows obvious lethal effect, and substantially suppresses the growth of tumor-bearing mice in-vivo tumour, and Jing is by Adr-
After GNs coupled complexes and siRap2b-GNs coupled complexes and illumination partner treatment, tumor size is saline control group
1/14, be used alone Adr-GNs coupled complexes matched group 1/8, be using Adr-GNs coupled complexes and illumination
The 1/2.6 of matched group.
Description of the drawings
Fig. 1 is the electromicroscopic photograph of the gold nanoshell that preparation is obtained.
Fig. 2 is the abosrption spectrogram of GNs, Adr-GNs coupled complex and siRap2b-GNs coupled complexes.
Fig. 3 is gold nanoshell（GNs）Wavelength be 808nm, power be 2w cm-2Laser irradiation under photo-thermal effect figure.
Fig. 4 is graph of a relation of the Adr-GNs coupled complexes medicine realeasing rate with the time for irradiating 1min per hour.
Fig. 5 is graph of a relation of the siRap2b-GNs coupled complexes medicine realeasing rate with the time for irradiating 1min per hour.
Fig. 6 is confocal laser scanning microscope tumor cell HCT116 to Adr-GNs coupled complexes and siRap2b-
The fluorescence contrast figure of GNs coupled complexes intake.
Fig. 7 is the impact comparison diagram of each drug on tumor cell HCT116 survival rates.
Fig. 8 is change comparison diagram of each test group to gross tumor volume before and after HCT116 treatment of colon cancer.
Fig. 9 is change comparison diagram of each test group to Mouse Weight before and after HCT116 treatment of colon cancer.
Figure 10 is the tumor object size contrast photo after each test group terminates to HCT116 treatment of colon cancer.
Specific embodiment
1st, gold nanoshell（GNs）Preparation：
The method of reference literature report（Jian You, Guodong Zhang, and Chun Li. Exceptionally
high payload of doxorubicin in hollow gold nanospheres for near-infrared
light-triggered drug release. ACS Nano, 2010. 4(2): 1033-1041.）, by 3.4 mg
AgNO3100 mL deionized waters are dissolved in 15 mg two citric acid monohydrate trisodiums, add 7.6 mg NaBH while stirring4, it is heated to
60 DEG C and stir 2 h.Room temperature is cooled to, 83.4 mg oxammonium hydrochloride .s are added, 5 min is stirred, is added 29.75 mg AgNO3,
Stir 2 more than h（Preferably overnight）.Then solution is heated to into 60 DEG C again, and rapidly joins the chlorine that 1 mL concentration is 20mg/mL
Auric acid aqueous solution, stopped reaction after stirring 1h in 60 DEG C.4 DEG C of 48 h of standing, discard bottom sediment.Upper solution is GNs, puts
Save backup in 4 DEG C of environment.By the method for the reports such as Prevo（Brian G. Prevo, Shelley A. Esakoff,
Alexander Mikhailovsky, and Joseph A. Zasadzinski. Scalable routes to gold
nanoshells with tunable sizes and response to near-infrared pulsed-laser
irradiation. Small, 2008. 4(8): 1183-1195）The concentration for determining GNs is 5 × 1010 mL-1。
Fig. 1 shows the electromicroscopic photograph of the GNs that preparation is obtained, it is seen that the diameter that GNs has hollow structure, GNs is about 28
nm。
2nd, the gold nanoshell of PEG modifications（GNs）Preparation：
In order to improve the stability of GNs structures, GNs is modified with PEG.
Take the GNs solution of 120 mL newly synthesis（Concentration is 5 × 1010 mL-1）, 14000 rpm be centrifuged 20 min, go
Clearly, 0.5 mL deionized waters are substantially soluble in, gained GNs solution concentrations are 1.2 × 1013 mL-1。
60mg mPEG-SH-2000 are weighed, deionized water is dissolved in and is settled to 10 mL, obtain 3 × 10-3 M mPEG-SH-
2000, deionized water presses 1:1000 are serially diluted to 3 × 10-9M, then take 50 μ l 3 × 10-9M mPEG-SH-2000 are molten
Liquid, is mixed with 950 μ L deionized waters, and gained mPEG-SH-2000 solution concentrations are 6 × 10-10 M。
0.5 mL 1.2 × 10 is added in 1.5 mL microcentrifugal tubes13 mL-1Gold nanoshell solution（Final concentration of 6 ×
1012 mL-1）With 0.5 mL 6 × 10-10M mPEG-SH-2000 solution（Final concentration of 3 × 10-10M）, mix, be stirred at room temperature
24 h, the terminating reaction after mPEG-SH-2000 is fully combined with GNs.14000 rpm are centrifuged 20 min, not anti-in removing supernatant
The mPEG-SH-2000 for answering, is precipitated with deionized water wash（That is, the GNs of PEG modifications）2 times, and be resuspended in deionized water, shape
Into the GNs solution of PEG modifications, its concentration is 6 × 1012 mL-1。
Made by taking, the GNs solution UV-visible-near infrared absorption instrument of PEG modifications scans its absorption spectrum, sees
Shown in Fig. 2.
3rd, the preparation of Adr-GNs coupled complexes：
Take 0.35 mg Adr and be added to 1 mL and contain 6 × 1012In the GNs solution of the PEG modifications of individual nanoparticle, it is stirred at room temperature
24h, the active amino and GNs for making Adr form coordinate bond.After reaction terminates, 14000 rpm are centrifuged 20 min.Reaction is produced
Washing of precipitate after be dried, obtain final product Adr-GNs coupled complexes.
Use dimethyl sulfoxide（DMSO）The Adr being coupled on eluting dissolving GNs, to Adr direct quantitatives, calculates the connection of Adr
Efficiency.
On the other hand unreacted Adr concentration in supernatant is determined, and the joint efficiency of Adr is calculated with indirect method.
Both the above method shows that the bonding ratio of Adr is 6 × 104Adr/GNs（That is, average each GNs particle loads 6
×104Individual Adr molecules）.
After the synthesis of Adr-GNs coupled complexes, its absorption spectrum is scanned with UV-visible-near infrared absorption instrument, seen
Shown in Fig. 2.
4th, the preparation of siRap2b-GNs coupled complexes：
The coupling process of the GNs and siRap2b of PEG modifications：
According to the siRNA sequence of Rap2b gene orders design（siRap2b）For 5 '-GACGAGCUAUUUGCCGAGATT-3 '.
GNs is connected to by 5 ' the end sulfydryls of SH-5 '-siRap2b-3 '-Rh123：0.5 mL is prepared with DEPC water
GNs（Wherein contain 6 × 1012Individual GNs particles）Mix with 40 μ g siRap2b, and 1 mL is settled to DEPC water, in room temperature bar
24 h of stirring reaction under part.
After reaction terminates, 14000 rpm are centrifuged 20 min, determine unreacted siRNA concentration in supernatant, use indirect method meter
Calculate siRap2b joint efficiencies.As a result show：The joint efficiency of siRap2b is 200/1（I.e. average each gold nanoshell particles are filled
Carry 200 siRNA molecules）.
After the synthesis of siRap2b-GNs coupled complexes, its absorption spectrum is scanned.
Fig. 2 respectively illustrates the absorption spectrum of GNs, Adr-GNs coupled complex and siRap2b-GNs coupled complexes
Figure, it is seen then that the characteristic absorption peak of the GNs solution with 786 nm of PEG modifications made by above step 2；What step 3 was obtained
Being coupled in Adr-GNs coupled complexes has Adr；And being coupled in the siRap2b-GNs coupled complexes of step 4 synthesis has
siRap2b。
5th, detect the photo-thermal effect of GNs particles：
By 1012Individual GNs particles are placed in the EP pipes containing 2 mL McCoy ' s 5A culture medium, and EP pipes are placed in 37 DEG C of water
In bath, with McCoy ' s 5A culture medium as control, the laser light for imposing 808 nm wavelength is shone（Optical density is 2 W cm-2）, swashing
The different time points of light illumination, real-time monitoring solution temperature draw time-temperature curve, obtain the photo-thermal effect comparison diagram shown in Fig. 3.
As seen from Figure 3：When laser irradiates 1 min, solution temperature is 44.7 DEG C.The temperature that the temperature releases the drug required by experiment in GNs
In the range of degree（41℃-45 ℃）, therefore the laser light of 808 nm wavelength can be shone（Optical density is 2 W cm-2）1 min conducts
The treatment conditions of follow-up drug release experiment, tumor cell killing experiments in vitro and interior therapeutic experiment.
6th, drug release experiment：
Drug release experiment is carried out under 37 DEG C of water-baths.
By 2 mL Adr-GNs or siRap2b-GNs solution（1012Individual nanoparticle/mL）It is sub-packed in EP pipes, puts 37 DEG C
Water-bath.All samples are divided into into two groups：Light group and non-light group.For non-light group, its load at 37 DEG C is mainly investigated
Drug stabilisation, respectively after 1 h of water-bath, 2 h, 3 h, by solution centrifugal（14000 rpm, 20 min), in detecting respective supernatant
Adr the or siRap2b concentration of release, so as to draw the drug release/time graph of Adr or siRap2b.And light group is then in 37 DEG C of water
Impose 1 min laser light photograph when bathing 1 h, 2 h, 3 h respectively（808 nm, 2 W cm-2）, caused with investigating short time illumination
Drug release profiles hop.
As a result as shown in Figure 4,5.
Fig. 4 shows graph of a relation of the Adr-GNs coupled complexes medicine realeasing rate with the time for irradiating 1min per hour.By Fig. 4
It can be seen that：Under conditions of the irradiation of no laser, prolongation over time, the medicine realeasing rate of Adr-GNs coupled complexes are gradually increasing,
In 37 DEG C of 1 h of water-bath, 2 h and 3 h, its medicine realeasing rate respectively 3.73%, 6.63% and 10.07%, but 1 min laser light of Jing
According to（808 nm, 2 W cm-2）Afterwards, the medicine realeasing rate of corresponding time point rises to 14.77%, 23.37% and 27.93% respectively.
Fig. 5 shows graph of a relation of the siRap2b-GNs coupled complexes medicine realeasing rate with the time for irradiating 1min per hour.By
Fig. 5 is visible：Under conditions of the irradiation of no laser, the medicine realeasing rate of siRap2b-GNs coupled complexes with the time prolongation gradually
Rise, in 37 DEG C of 1 h of water-bath, 2 h and 3 h, its medicine realeasing rate is respectively 3.47%, 5.87% and 9.23%, but 1 min of Jing swash
Light illumination（808 nm, 2 W cm-2）Afterwards, the medicine realeasing rate of corresponding time point rises to 13.07%, 20.90% and 28.27% respectively.
7th, cellular uptake experiment：
By human colon carcinoma HCT116 cells copolymerization Jiao plate in cultivate 12h after, be separately added into Adr-GNs coupled complexes and
SiRap2b-GNs coupled complexes, final concentration are 3 × 1011Individual nanoparticle/mL.37 DEG C be incubated respectively 0 h, 2 h, 4 h,
6 h, 12 h, suction abandon culture fluid, and use PBS（pH 8.0）Washing 2 times, then observes Adr under Laser Scanning Confocal Microscope
And Rhodamine 123（Rh123）（Hold positioned at siRap2b 3 '）Produced cell fluorescence, as shown in Figure 6.
As seen from Figure 6：Cell fluorescence intensity produced by prolongation over time, Adr and Rh123 present first rise, after
The overall trend of decline, and maximum is reached when 4 h is incubated for 37 DEG C.
8th, the impact experiment to tumor cell survival：
With McCoy ' the s 5A complete mediums containing 10% hyclone in 37 DEG C, 5% CO2HCT116 is cultivated in incubator thin
Born of the same parents, when cell growth is to covering culture dish about 80%, is digested with 0.25% trypsin-EDTA, and are prepared with complete medium
Into single cell suspension, 96 orifice plates are laid on 3000 cells/wells, put 37 DEG C, 5% CO2Cultivate in incubator, use instead after 24 h and contain
The low serum free culture system liquid of 1% hyclone.All cells are divided into 7 groups, per group of 6 multiple holes.
Group 1：The final concentration of Adr-GNs+siRap2b-GNs+ illumination, wherein Adr-GNs be respectively 0.0357,0.0179,
The final concentration respectively 1.25 of the 0.0089285th, 0.00446,0.00223,0.00112 and 0.000558 nM, loading Adr,
0.625th, 0.3125,0.15625,0.078125,0.0390625 and 0.01953125 μ g/mL, and in variable concentrations Adr-GNs
Final concentration of 5 × 10 are added in medicine feeding hole-4 The siRap2b-GNs of nM（Final concentration of 0.1 nM of siRap2b of loading）, 37
After DEG C being altogether incubated 4h, irradiate 1 minute per hole laser（808 nm, 2 W cm-2）, it is further continued for being incubated 44h.
Group 2：Adr-GNs+siRap2b-GNs, dosing are identical with group 1, are incubated 48 h, no light altogether.
Group 3：Adr-GNs+ illumination, wherein Adr-GNs final concentration of 0.0357,0.0179,0.0089285,
0.00446th, 0.00223,0.00112,0.000558 nM, load Adr final concentration be respectively 1.25,0.625,0.3125,
0.15625th, 0.078125,0.0390625,0.01953125 μ g/mL, and add in variable concentrations Adr-GNs medicine feeding holes
Final concentration of 5 × 10-4 The GNs of the unloaded medicine of nM so as to which nanoparticle sum is identical with group 1.After 37 DEG C are incubated 4 h altogether,
Irradiate 1 minute per hole laser（808 nm, 2 W•cm-2）, it is further continued for being incubated 44 h.
Group 4：Adr-GNs, dosing are identical with group 3, are incubated 48 h, no light altogether.
Group 5：Only Adr, its final concentration be respectively 1.25,0.625,0.3125,0.15625,0.078125,0.0390625,
0.01953125 μ g/mL, are incubated 48 h altogether.
Group 6：Adr+siRap2b, Adr final concentration be respectively 1.25,0.625,0.3125,0.15625,0.078125,
0.0390625th, 0.01953125 μ g/mL, and add final concentration of 0.1 nM's in all variable concentrations Adr medicine feeding holes
siRap2b。
Group 7：Adr+siRap2b-lip2000, Adr final concentration be respectively 1.25,0.625,0.3125,0.15625,
0.078125th, 0.0390625,0.01953125 μ g/mL, and add and transfection reagent in all variable concentrations Adr medicine feeding holes
The siRap2b, final concentration of 0.1 nM of siRap2b of the premixs of Lipofectamine 2000.
With not dosing cell as negative control, while arranging blank（Cell is not added with, remaining operation all same）.With medicine
Thing adds 10 μ l MTT per hole after being incubated altogether, continues at 37 DEG C, 5% CO2Cultivate in incubator, inhale after 4 h and abandon culture medium, per hole
Plus 150 μ l DMSO colour developing, at 490 nm wavelength of microplate reader survey light absorption value.Cell proliferation inhibition rate=（A samples-A is empty）/（A
P- A is empty）* 100%.
As a result it is as shown in Figure 7.As seen from Figure 7：SiRap2b can significantly improve the effect of Adr killing tumor cells, and group
1（Adr-GNs+siRap2b-GNs+ illumination）Fragmentation effect it is best.
9th, tumor-bearing mice Experiment on therapy：
By HCT116 cells in 37 DEG C, 5% CO2Cultivate in incubator, when cell growth is to covering culture dish about 80%, use
0.25% trypsin-EDTA digests, and is washed 2 times using ice-cold McCoy ' s 5A serum-free mediums, is resuspended in serum-free
In culture medium（1.25×106/mL）, in mice oxter subcutaneous vaccination HCT116 tumor cells（2.5×106Cell/200 μ l/
Only）.Wherein 36 tumor sizes are selected after 6 days（Diameter is about 3-4 mm）, body weight it is close（20 g or so）Transplanted tumor mice,
It is divided into 6 groups, 6 per group.
Experimental group 1（Adr-GNs+siRap2b-GNs+ illumination）：GNs（0.0857 nmol/kg, the Adr of loading is 3 mg/
kg）+siRap2b-GNs（1.25 nmol/kg, the siRap2b of loading is 0.25 μm of ol/kg）, after tail vein is administered 6 h, light
According to 1 min（808 nm, 2 W•cm-2）.
Experimental group 2（Adr-GNs+siRap2b-GNs）：, no light identical with test group 1 is administered.
Experimental group 3（Adr-GNs+ illumination）：GNs（0.0857 nmol/kg, the Adr of loading is 3 mg/kg）+ unloaded medicine
The GNS of thing（1.25 nmol/kg so as to identical with 1 nanoparticle sum of test group）, after tail vein is administered 6 h, 1 min of illumination
（808 nm, 2 W•cm-2）.
Experimental group 4（Adr-GNs）：, no light identical with test group 3 is administered.
Experimental group 5（Only Adr）：Adr（3 mg/kg）.
Experimental group 6（Normal saline）：Administration normal saline, as control.
A medicine was given during experiment per 3 days（Administered volume is less than 200 μ l every time）, while measuring diameter of tumor and little
Mus body weight, according to formula V=ab2/ 2 calculate gross tumor volume（A is tumor major diameter, and b is tumor minor axis）, tumor growth curve is drawn,
Tumour inhibiting rate is calculated, body weight change is observed.It is administered 5 times altogether, after 15 days, puts to death mice, peel off tumor and take pictures, tumor is stored in Fu Er
Malin's solution（-70 ℃）.
Fig. 8 shows change comparison diagram of each test group to gross tumor volume before and after HCT116 treatment of colon cancer.Can by Fig. 8
See：Adr substantially suppresses the growth of HCT116 tumors, siRap2b further enhance the Tumor growth inhibition effect of Adr, and
Experimental group 1（Adr-GNs+siRap2b-GNs+ illumination）Tumor growth inhibitory effect it is best.
Fig. 9 shows change comparison diagram of each test group to Mouse Weight before and after HCT116 treatment of colon cancer.Can by Fig. 9
See：Jing after Adr treatments, each experimental mice body weight is apparently higher than saline control group（Experimental group 6）, and experimental group 1
（Adr-GNs+siRap2b-GNs+ illumination）Average mice body weight apparently higher than other experimental grouies, illustrate 1 Mice Body of experimental group
, Jing after treatment, the impact to Mouse Weight is minimum for interior tumor.
Tumor object size after each test group that Figure 10 shows terminates to HCT116 treatment of colon cancer contrasts photo.By scheming
10 is visible：Jing after Adr treatments, HCT116 tumor mass reductions, and siRap2b can strengthen the therapeutic effect of Adr, further reduce
Gross tumor volume, and Jing experimental grouies 1（Adr-GNs+siRap2b-GNs+ illumination）Mouse tumor average external volume after treatment is minimum,
For saline control group（Experimental group 6）1/14, experimental group 5（Only Adr）1/8, experimental group 3（Adr-GNs+ illumination）1/
2.6, therefore the effect of the therapeutic scheme is best.
<110>Yangzhou University
<120>Adr-GNs coupled complexes for targeted therapy of malignant and preparation method thereof
<160>1
<210>1
<211>21
<212>DNA
<213>Artificial sequence
<220>
<223>According to the siRNA sequence of Rap2b gene orders design
<400>1
gacgagcuauuugccgagat t。
<110>Yangzhou University
<120>Adr-GNs coupled complexes for targeted therapy of malignant and preparation method thereof
<160>1
<210>1
<211>21
<212>DNA
<213>Artificial sequence
<220>
<223>According to the siRNA sequence of Rap2b gene orders design
<400>1
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Claims (2)
1. the Adr-GNs coupled complexes of targeted therapy of malignant are used for, it is characterised in that：In the gold nanoshell of PEG modifications
Upper coupling Adr.
2. the preparation method of the Adr-GNs coupled complexes of targeted therapy of malignant, its feature are used for as claimed in claim 1
It is to comprise the following steps：
1）Gold nanoshell solution and mPEG-SH-2000 mixing are reacted, the gold nanoshell of PEG modifications is obtained；
2）The gold nanoshell solution mixing that Adr and PEG is modified is reacted, and Adr-GNs coupled complexes are obtained.
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Citations (1)
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| CN102961337A (en) * | 2012-12-14 | 2013-03-13 | 哈尔滨工业大学 | Preparation method of target compound nano particle |
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| CN102961337A (en) * | 2012-12-14 | 2013-03-13 | 哈尔滨工业大学 | Preparation method of target compound nano particle |
Non-Patent Citations (2)
| Title |
|---|
| JIAN YOU ET AL.: "Exceptionally High Payload of Doxorubicin in Hollow Gold Nanospheres for Near-Infrared Light Triggered Drug Release", 《ACSNANO》 * |
| XINYUE ZHANG ET AL: "Rap2b, a novel p53 target, regulates p53-mediated pro-survival function", 《CELL CYCLE》 * |
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