CN108273073A - Extra small functionalized nano cluster, nano-probe and its preparation method and application - Google Patents
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0002—General or multifunctional contrast agents, e.g. chelated agents
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- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
- A61K49/0056—Peptides, proteins, polyamino acids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0063—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres
- A61K49/0069—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form
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- A61K49/0091—Microparticle, microcapsule, microbubble, microsphere, microbead, i.e. having a size or diameter higher or equal to 1 micrometer
- A61K49/0093—Nanoparticle, nanocapsule, nanobubble, nanosphere, nanobead, i.e. having a size or diameter smaller than 1 micrometer, e.g. polymeric nanoparticle
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- A—HUMAN NECESSITIES
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- A61K49/04—X-ray contrast preparations
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
- A61K49/10—Organic compounds
- A61K49/14—Peptides, e.g. proteins
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
- A61K49/1818—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles
- A61K49/1821—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles
- A61K49/1824—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles
- A61K49/1827—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle
- A61K49/1866—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle the nanoparticle having a (super)(para)magnetic core coated or functionalised with a peptide, e.g. protein, polyamino acid
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Abstract
The present invention provides a kind of extra small functionalized nano cluster, the use of GSH is template, by the sulfydryl on the surfaces GSH, carboxyl and Au atoms, gadolinium Gd atoms huge legendary turtle and, form the internal nanometer clustering architecture for containing two kinds of atoms simultaneously;The extra small functionalized nano probe of cNGR in surface modification is also provided, and provide preparation method, because it is with the tumor neovasculature ability of specific recognition, and fluorescence is generated by the quantum confined effect of Au atoms and to the absorption of X-ray absorption coefficient, realizes fluorescence imaging and CT imagings;Shorten the longitudinal relaxation time (T1) of free water proton by Gd atoms, Enhanced MR contrast is, it can be achieved that application in tumor neogenetic blood vessels multi-modality imaging.Nano-particles size small (< 10nm) prepared by the present invention, good biocompatibility, toxicity are low;Using the modularization assembling performance of nano material, the efficient collaboration of multiple functions is realized;Yield height is prepared, is suitble to produce in enormous quantities.
Description
Technical field
The present invention relates to nanosecond medical science fields, and in particular to a kind of extra small functionalized nano cluster, nano-probe and its preparation
Methods and applications.
Background technology
Cancer Mortality and the death rate increase year by year, and the early diagnosis of tumour and personalized treatment are to improve patient's life
Deposit rate most efficient method.But most of patients has arrived middle and advanced stage when making a definite diagnosis, and tumour has shifted, when missing optimal treatment
Machine.The accurate evaluation of tumor-microvessel has evaluation tumour progression, formulation personalized therapy program and prediction prognosis important
Clinical meaning.However, clinically existing contrast agent (the sour sodium of gadolinium spray, Iohexol etc.) lacks targeting, susceptibility difference and nothing
The live body capilary of 300 μm of method direct image diameter < causes conventional image technology to be difficult to dynamic, accurately assess tumor vessel
It is newborn.In addition to this, the mechanism of Tumor Angiongesis is complicated, there are individual difference between different patients, single imaging pattern without
Method obtains all information of lesions position, causes assessment result inaccurate.Therefore, in order to improve tumor neogenetic blood vessels imaging standard
True property realizes that the targeting of imaging and high sensitivity are just of great significance.
Multi-modality imaging based on nano meter biomaterial can be synchronized in conjunction with a variety of Imaging Technologies, be realized between each pattern
Mutual supplement with each other's advantages, to achieve the purpose that improve imaging sensitivity and resolution ratio.Currently, making nano material that there are a variety of imaging moulds
The method of state mainly assembles different image-forming components, but nano material grain size would generally be caused largely to increase
Greatly, easily most nano material is made to enter liver, spleen by the capture of endothelium network later into blood circulation inside body system
Equal tissues cause only a small amount of nano material that can reach cancer target region, and targeting efficiency is low, affects due imaging
Effect.
The nano metal cluster of functionalization is in a manner of biomineralization, and the molecule being made of precious metal elements such as Au, Ag gathers
Collective is usually made of several to dozens of atoms, and the diameter of core is less than 5nm, and size and physicochemical property are between single atom
Between nano-particle.Such as gold atom quantum confined effect and X-ray absorption, can send out stronger fluorescence, enhanced CT at
Picture and radio therapy sensitization effect.A kind of widely applied nano material is increasingly becoming in many fields of nanosecond science and technology in recent years.
Invention content
The purpose of the present invention is be used for present in Tumor Angiongesis imaging in view of current multi-modal functionalized nano material
Defect so that it is preferably applied for tumor vessel in order to improve the targeting efficiency and image contrast of multi-modal nano-probe
Imaging is generated, a kind of extra small (< with endothelial cells in tumor neogenetic blood vessels targeting, MR/CT/ fluorescence multi-modality imagings is provided
10nm) functionalized nano cluster, nano-probe and preparation method thereof.
The technical scheme is that:
The nano-cluster synthesis template of the present invention selects glutathione (glutathione-glutamyl cysteingl-
Glycine, GSH), the small peptide being made of glutamic acid, cysteine and glycine contains three kinds of amino, carboxyl and sulfydryl
Functional group can be chelated the metal ions such as gold, gadolinium, and have good hydrophily, be widely used in conjugated biological molecules
Prepare the contrast agent etc. of pharmaceutical carrier, nanoscale.
The extra small functionalized nano cluster of the present invention, that is, it is template to select GSH, utilizes the sulfydryl on the surfaces GSH, carboxyl and gold
(Au) atom, gadolinium (Gd) atom huge legendary turtle and, forming nano-cluster simultaneously keeps it internal while containing two kinds of atoms, passes through the quantum of Au atoms
Confinement effect generates fluorescence and the absorption to X-ray absorption coefficient, realizes fluorescence imaging and CT imagings;Shortened by Gd atoms
Destination organization t1 values, Enhanced MR contrast.
Angiogenesis frizzled receptor aminopeptidase (CD13) is a kind of closely related with angiogenesis and tumor invasion
Transmembrane protein, height are expressed in endothelial cells in tumor neogenetic blood vessels and tumor cell surface, such as liver cancer, oophoroma, breast cancer, uterus
The malignant tumours such as endometrial carcinomas, brain tumor, adenocarcinoma of lung, brain tumor.Targeting preparation used in the present invention is cyclic n nitroso compound GR (cNGR, cyclic
NAc-Cys-Asn-Gly-Arg-Cys-Gly-Gly-Lys (Ac)-NH2, Mr=930.05), which is to pass through phage display technology
Show the ligands specific that technology screening goes out, can efficiently be specifically bound with CD13.Currently, NGR targeted drugs have been enter into
Clinical experimental stage has broad application prospects, but small-molecule drug there are still tachytrophism, blood circulation is poor the defects of.
The specific recognition of CD13 is acted in the height expression of malignant tumour neovascular endothelium cell and cNGR using CD13, can be set
The nano-probe of meter label cNGR, target tumor neovascular endothelium cell are imaged, and the non-spy of normal tissue is reduced
Opposite sex aggregation, enhances tumor neovasculature imaging effect.
The extra small functionalized nano probe of the present invention passes through the extra small functionalized nano cluster surface modification obtained by aforementioned
The nano-probe with the tumor neovasculature ability of specific recognition is prepared in upper cNGR.
Specifically preparation method is:In molar ratio by chlorauric acid solution and gadolinium chloride solution, glutathione solution:1:(1~
10):(4~6) are placed in single-necked flask, (1000rpm) are mixed slowly under water bath condition, temperature is risen to 70 DEG C after a certain period of time
To form nano-cluster.Centrifugation 20min removes supernatant afterwards for 24 hours for reaction, is used in combination 10m1 deionized waters to wash 3 times, obtains final product
Extra small functionalized nano cluster.
Then by cNGR polypeptides and nano-cluster according to mass volume ratio 1:(30~300) Bioconjugation is carried out:It will receive first
Rice cluster and 10 μ L EDC (8mg/mL) and 20 μ L NHS (8mg) be mixed in 1mL phosphate buffers (PBS buffer, 1 ×,
PH 7.4) in, cNGR polypeptides, the reaction was continued 2h is added under 37 DEG C of environment after concussion reaction 20min, centrifugal purification is simultaneously stored in
In PBS buffer solution (1 ×, pH 7.4), obtain extra small functionalized nano probe.
Reagent and drug used in the present invention can pass through commercially available acquisition.
Extra small functionalized nano probe of the present invention, can be applied to tumor neogenetic blood vessels multi-modality imaging, have such as
Lower advantageous effect:Nano-particles size small (< 10nm), good biocompatibility, the toxicity of preparation are low;Utilize the mould of nano material
Block assembling property realizes the efficient collaboration of multiple functions;Yield height is prepared, is suitble to produce in enormous quantities;CNGR is modified more
Mode nano-probe can effectively improve it to tumor neovasculature targeting ability, and imaging can be effectively improved in conjunction with multi-modality imaging
Sensitivity and resolution ratio, improve diagnostic.
Description of the drawings
Fig. 1:Extra small functionalized nano probe structure schematic diagram
Fig. 2:The ultra-violet absorption spectrum of multi-modal extra small nano-probe
Fig. 3:The particle diameter distribution of multi-modal extra small nano-probe
Fig. 4:The fluorescence emission spectrum of multi-modal extra small nano-probe
Fig. 5:MR the and CT values of the multi-modal extra small nano-probe of various concentration:Wherein (A) MR images;(B) corresponding MR
Value;(C) CT images;(D) corresponding CT values
Fig. 6:The aggregation (silver staining) of multi-modal extra small nano-probe chrotoplast within the tumor
Specific implementation mode
Extra small functionalized nano probe structure schematic diagram of the present invention as shown in Figure 1, its using GSH be template, lead to
Cross sulfydryl, carboxyl and the Au atoms on the surfaces GSH, gadolinium Gd atoms huge legendary turtle and, form the internal extra small function of containing two kinds of atoms simultaneously
Change nano-cluster;The cNGR in the extra small functionalized nano cluster surface modification.
Preparation method is as follows:
Embodiment 1:
By 2ml chlorauric acid solutions (a concentration of 24mM) and 1ml gadolinium chlorides solution (48mM), 24mL glutathione solutions
(4mM) is placed in the single-necked flask of 100mL, rises to temperature after mixing slowly (1000rpm) 20min under the conditions of water-bath (30 DEG C)
70 DEG C, to form nano-cluster, are reacted for 24 hours centrifugation 20min removals supernatant afterwards, are used in combination 10mL deionized waters to wash 3 times, are received
Rice cluster.
30 μ L nano-clusters (50mg/mL), 10 μ L EDC (8mg/mL) and 20 μ L NHS (8mg) are mixed in 1mL phosphate
Buffer solution (PBS buffer, 1 ×, pH 7.4) in, 5 μ L cNGR polypeptides are added under 37 DEG C of environment after concussion reaction 20min
(1mg/mL), the reaction was continued 2h, centrifugal purification are simultaneously stored in PBS buffer solution (1 ×, pH 7.4), obtain extra small functionalization and receive
Rice probe.
Gained nano-probe is diluted to 0.5mg/mL, its UV absorption light is detected using ultraviolet-visible spectrophotometer
Spectrum, detects its grain size using laser particle analyzer, its fluorescence emission spectrum is detected using sepectrophotofluorometer.
As shown in Fig. 2 ultra-violet absorption spectrums, nano-probe has absorption peak specific to typical nanogold cluster, illustrates me
Successfully prepared extra small functionalized nano probe for tumor neogenetic blood vessels multi-modality imaging.
As Fig. 3 particle diameter distributions indicate that the grain size of nano-probe is about 4nm, it was demonstrated that prepared nano material size belongs to super
Small range (< 10nm).
As shown in Fig. 4 fluorescence emission spectrums, which can launch the glimmering of 600nm or so under burst of ultraviolel
Light.
Embodiment 2:
By 2ml chlorauric acid solutions (a concentration of 24mM) and 1ml gadolinium chlorides solution (120mM), 24mL glutathione solutions
(4mM) is placed in the single-necked flask of 100mL, rises to temperature after mixing slowly (1000rpm) 20min under the conditions of water-bath (30 DEG C)
70 DEG C to form nano-cluster.Centrifugation 20min removes supernatant afterwards for 24 hours for reaction, is used in combination 10mL deionized waters to wash 3 times, is received
Rice cluster.
30 μ L nano-clusters (50mg/mL), 10 μ L EDC (8mg/mL) and 20 μ L NHS (8mg) are mixed in 1mL phosphate
Buffer solution (PBS buffer, 1 ×, pH 7.4) in, 10 μ L cNGR polypeptides are added under 37 DEG C of environment after concussion reaction 20min
(1mg/mL), the reaction was continued 2h, centrifugal purification are simultaneously stored in PBS buffer solution (1 ×, pH 7.4).Preparation-obtained nanometer
Probe grain size is 6nm.
Case study on implementation 3
By 2ml chlorauric acid solutions (a concentration of 24mM) and 1ml gadolinium chlorides solution (120mM), 24mL glutathione solutions
(5mM) is placed in the single-necked flask of 100mL, rises to temperature after mixing slowly (1000rpm) 20min under the conditions of water-bath (30 DEG C)
70 DEG C to form nano-cluster.Centrifugation 20min removes supernatant afterwards for 24 hours for reaction, is used in combination 10mL deionized waters to wash 3 times, is received
Rice cluster.
30 μ L nano-clusters (50mg/mL), 10 μ L EDC (8mg/mL) and 20 μ L NHS (8mg) are mixed in 1mL phosphate
Buffer solution (PBS buffer, 1 ×, pH 7.4) in, 15 μ L cNGR polypeptides are added under 37 DEG C of environment after concussion reaction 20min
(1mg/mL), the reaction was continued 2h, centrifugal purification are simultaneously stored in PBS buffer solution (1 ×, pH 7.4).Preparation-obtained nanometer
Probe grain size is 3.8nm.
Embodiment 4:
By 2ml chlorauric acid solutions (a concentration of 24mM) and 1ml gadolinium chlorides solution (120mM), 24mL glutathione solutions
(6mM) is placed in the single-necked flask of 100mL, rises to temperature after mixing slowly (1000rpm) 20min under the conditions of water-bath (30 DEG C)
70 DEG C to form nano-cluster.Centrifugation 20min removes supernatant afterwards for 24 hours for reaction, is used in combination 10mL deionized waters to wash 3 times, is received
Rice cluster.
30 μ L nano-clusters (50mg/mL), 10 μ L EDC (8mg/mL) and 20 μ L NHS (8mg) are mixed in 1mL phosphate
Buffer solution (PBS buffer, 1 ×, pH 7.4) in, 30 μ L cNGR polypeptides are added under 37 DEG C of environment after concussion reaction 20min
(1mg/mL), the reaction was continued 2h, centrifugal purification are simultaneously stored in PBS buffer solution (1 ×, pH 7.4).Preparation-obtained nanometer
Probe grain size is 5.04nm.
It is diluted to the concentration range of 0-1mg/mL, is placed in GE 3.0T superconducting magnetic resonances scanning machines and 64 row's spirals
In CT machines, scanning grey pictures simultaneously carry software reading gray value with instrument, itself MR and CT imaging function, such as Fig. 5 is detected with this
Shown, MR and CT values all increase with the raising of solution concentration, illustrate the ability that the nano-probe has multi-modality imaging.
Embodiment 5:
By 2ml chlorauric acid solutions (a concentration of 24mM) and 1ml gadolinium chlorides solution (240mM), 24mL glutathione solutions
(6mM) is placed in the single-necked flask of 100mL, rises to temperature after mixing slowly (1000rpm) 20min under the conditions of water-bath (30 DEG C)
70 DEG C to form nano-cluster.Centrifugation 20min removes supernatant afterwards for 24 hours for reaction, is used in combination 10mL deionized waters to wash 3 times, is received
Rice cluster.
30 μ L nano-clusters (50mg/mL), 10 μ L EDC (8mg/mL) and 20 μ L NHS (8mg) are mixed in 1mL phosphate
Buffer solution (PBS buffer, 1 ×, pH 7.4) in, 50 μ L cNGR polypeptides are added under 37 DEG C of environment after concussion reaction 20min
(1mg/mL), the reaction was continued 2h, centrifugal purification are simultaneously stored in PBS buffer solution (1 ×, pH 7.4).Preparation-obtained nanometer
Probe grain size is 6.32nm.
Female balb/c mouse keep the feeding of constant temperature (20 DEG C -26 DEG C) and constant humidity (50%-65%) condition without specific disease
In simple laminar-flow rack.Every mouse right shoulder inoculates 200 μ L PBS cell suspensions (cell content is 1 × 107), tumour
Grow to 80-100mm3Afterwards, by the nano-probe of 200 a concentration of 20mg/mL of μ L by tail vein injection to mice with tumor body, and one
After fixing time take out tumor tissues carry out silver staining experiment with prove nano-probe Tumor Angiongesis region enrichment condition.Knot
Fruit illustrates the nanometer as shown in fig. 6, more concentratedly occur a large amount of nano-probe on the endothelial cell of Tumor Angiongesis
Probe can specifically be targeted to tumor neogenetic blood vessels position.
One embodiment of the present invention has been described in detail above, but the content be only the present invention preferable implementation
Example should not be construed as limiting the practical range of the present invention.It is all according to all the changes and improvements made by the present patent application range
Deng should all still fall within the scope of the patent of the present invention.
Claims (4)
1. a kind of extra small functionalized nano cluster, it is characterised in that:Using GSH be template, by the sulfydryl on the surfaces GSH, carboxyl with
Au atoms, gadolinium Gd atoms huge legendary turtle and, form the internal extra small functionalized nano cluster for containing two kinds of atoms simultaneously.
2. a kind of extra small functionalized nano probe, it is characterised in that:The use of GSH is template, passes through the sulfydryl on the surfaces GSH, carboxyl
With Au atoms, gadolinium Gd atoms huge legendary turtle and, form the internal extra small functionalized nano cluster for containing two kinds of atoms simultaneously;In the extra small work(
CNGR in nano-cluster surface modification can be changed, the extra small functionalization with specific recognition tumor neogenetic blood vessels ability is prepared and receives
Rice probe.
3. the preparation method of the extra small functionalized nano probe described in claim 2, it is characterised in that:Include the following steps:
In molar ratio by chlorauric acid solution and gadolinium chloride solution, glutathione solution:1:(1~10):(4~6) are placed in single port burning
In bottle, (1000rpm) is mixed slowly under water bath condition, temperature is risen to 70 DEG C to form nano-cluster after a certain period of time, reaction is for 24 hours
Centrifugation 20min removes supernatant afterwards, is used in combination 10m1 deionized waters to wash 3 times, obtains extra small functionalized nano described in claim 1
Cluster;
By cNGR polypeptides and nano-cluster according to mass volume ratio 1:(30~300) Bioconjugation is carried out:First by nano-cluster and 10 μ
L EDC (8mg/mL) and 20 μ L NHS (8mg) be mixed in 1mL phosphate buffers (PBS buffer, 1 ×, pH 7.4) in,
CNGR polypeptides, the reaction was continued 2h is added under 37 DEG C of environment after concussion reaction 20min, centrifugal purification is simultaneously stored in PBS buffer solution
(1 ×, pH 7.4), finally obtain extra small functionalized nano probe.
4. the extra small functionalized nano probe described in claim 2 is to the application in tumor neogenetic blood vessels multi-modality imaging.
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2020201812A1 (en) * | 2019-04-02 | 2020-10-08 | Wutthinitikornkit Yanee | Dynamic clusters of atom particles activated in extracellular fluid |
CN112816513A (en) * | 2020-12-30 | 2021-05-18 | 国家纳米科学中心 | Gold-gadolinium composite nano probe, preparation method and application |
CN113304283A (en) * | 2021-05-19 | 2021-08-27 | 吉林大学 | Au/Mn nanocluster, preparation method and application thereof in NIR/MRI/CT multi-mode imaging |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20090298115A1 (en) * | 2008-05-29 | 2009-12-03 | Chung Yuan Christian University | Fluorescent Gold Nanocluster and Method for Forming the Same |
US20110300532A1 (en) * | 2009-03-05 | 2011-12-08 | Wilhelm Jahnen-Dechent | Control of the toxicity of gold nanoparticles |
CN102366632A (en) * | 2011-08-22 | 2012-03-07 | 长春工业大学 | Paramagnetic metal complex functionalized fluorogold nano-cluster magnetic resonance and fluorescence imaging contrast agent |
CN104749151A (en) * | 2015-04-08 | 2015-07-01 | 东南大学 | Application of glutathione-based stable gold nano cluster particles to detection of sulfhydryl compound |
CN105363043A (en) * | 2014-08-08 | 2016-03-02 | 屈晓超 | RGD-labeled fluorescent gold nano-cluster preparation method |
-
2018
- 2018-04-19 CN CN201810352318.4A patent/CN108273073A/en active Pending
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20090298115A1 (en) * | 2008-05-29 | 2009-12-03 | Chung Yuan Christian University | Fluorescent Gold Nanocluster and Method for Forming the Same |
US20110300532A1 (en) * | 2009-03-05 | 2011-12-08 | Wilhelm Jahnen-Dechent | Control of the toxicity of gold nanoparticles |
CN102366632A (en) * | 2011-08-22 | 2012-03-07 | 长春工业大学 | Paramagnetic metal complex functionalized fluorogold nano-cluster magnetic resonance and fluorescence imaging contrast agent |
CN105363043A (en) * | 2014-08-08 | 2016-03-02 | 屈晓超 | RGD-labeled fluorescent gold nano-cluster preparation method |
CN104749151A (en) * | 2015-04-08 | 2015-07-01 | 东南大学 | Application of glutathione-based stable gold nano cluster particles to detection of sulfhydryl compound |
Non-Patent Citations (4)
Title |
---|
WENJUN LE, ET AL.: "Facile Synthesis of Gd-Functionalized Gold Nanoclusters as Potential MRI/CT Contrast Agents", 《NANOSMATERIALS》 * |
ZHENTAO LUO,ET AL.: "From Aggregation-Induced Emission of Au(I)−Thiolate Complexes to Ultrabright Au(0)@Au(I)−Thiolate Core−Shell Nanoclusters", 《JOURNAL OF AMERICAN CHEMICAL SOCIETY》 * |
曾戎编著: "《多糖基高分子-药物轭合物的》", 31 May 2011, 广州:华南理工大 * |
武明豪等: "cNGR功能化金纳米粒子探针在乳腺癌血管生成CT成像中的应用", 《国际生物医学工程杂志》 * |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2020201812A1 (en) * | 2019-04-02 | 2020-10-08 | Wutthinitikornkit Yanee | Dynamic clusters of atom particles activated in extracellular fluid |
CN114901261A (en) * | 2019-04-02 | 2022-08-12 | 淅川海灵生物科技有限公司 | Dynamic clustering of activated atomic particles in extracellular fluid |
CN112816513A (en) * | 2020-12-30 | 2021-05-18 | 国家纳米科学中心 | Gold-gadolinium composite nano probe, preparation method and application |
CN112816513B (en) * | 2020-12-30 | 2023-11-07 | 国家纳米科学中心 | Jin composite nano probe, preparation method and application |
CN113304283A (en) * | 2021-05-19 | 2021-08-27 | 吉林大学 | Au/Mn nanocluster, preparation method and application thereof in NIR/MRI/CT multi-mode imaging |
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