CN104758959A - Preparation method of radionuclide 131I-labeled folic acid targeted multifunctional dendrimer - Google Patents
Preparation method of radionuclide 131I-labeled folic acid targeted multifunctional dendrimer Download PDFInfo
- Publication number
- CN104758959A CN104758959A CN201510171048.3A CN201510171048A CN104758959A CN 104758959 A CN104758959 A CN 104758959A CN 201510171048 A CN201510171048 A CN 201510171048A CN 104758959 A CN104758959 A CN 104758959A
- Authority
- CN
- China
- Prior art keywords
- hpao
- peg
- dendrimer
- labelling
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Medicinal Preparation (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention relates to a preparation method of a radionuclide 131I-labeled folic acid targeted multifunctional dendrimer. The preparation method comprises the following steps: (1) adding EDC and an NH2-PEG-COOH solution to folic acid for reacting so as to obtain FA-PEG-COOH; (2) adding HPAO to a dendrimer solution for reacting so as to obtain G5.NH2-HPAO; (3) adding EDC to the FA-PEG-COOH solution, activating, and adding to the G5.NH2-HPAO water solution for reacting so as to obtain dendrimer solution; and (4) adding N(C2H5)3 and acetic anhydride to the solution for reacting so as to obtain a functionalized dendrimer solution, and then adding Na131I for carrying out a labeling reaction so as to obtain the multifunctional dendrimer. The material prepared by the invention has excellent SPECT imaging and radioactive therapy functions, and the material has a potential application value in the field of forming a nano material integrating diagnosis and treatment.
Description
Technical field
The invention belongs to the preparation field of functional dendrimer material, particularly a kind of radionuclide
131the multi-functional tree-shaped macromolecular preparation method of the folate-targeted of I labelling.
Background technology
Cancer be body under various carcinogenic factor effect, the cell of local organization loses the normal regulation to its growth on gene level, causes its clonal abnormality hypertrophy and the abnormality that formed.In view of local infiltration and the far-end transfer characteristic of malignant tumor, make the period of the therapeutic effect of cancer and medical expense and discovery cancer closely related.Discovery and early treatment reduce cancer mortality effective method the most early.Along with the development of nanometer science and technology, various nanometer system has started the early diagnosis and therapy being applied to cancer, comprise ferric oxide nanometer particle, quantum dot, CNT, gold nano grain, dendrimer, silicon nano materials etc. all can be endowed imaging and treatment function, and desirable nanoparticle systems that can be used in the early stage targeting diagnosis of cancerous cell and treatment should the targeted molecular of load simultaneously, imaging agents molecule and medicine, both enough preparations and medicine can be carried, again can by its efficiently orientation be transported to the focal area such as tumor, to provide sufficient imaging signal and drug level to focal zone.Therefore, Clinics and Practices combines by the nano load system by functionalization, is expected " diagnosis and treatment " integration realizing cancer.
Medicine imaging technique be by collecting, display radionuclide distribution density in vivo and flowing situation of change, obtain a kind of technology of the shape of internal organs, skeleton and pathological changes, position and function information.Radioactive isotope reagent is injected by blood vessel and assemble in vivo with blood flow, distribute, time to time change.Therefore, nuclear medicine is beyond the scope of anatomical structure development, the changes of function of internal organs or system can be reflected, and can effective diagnosing tumor be carried out, tumor can be found easily in the development of Whole body bone scan, thyroid and parathyroid function, liver blood pond in developing especially and differentiate the character of tumor.The probe of normal use uses
18f,
13n,
11c,
99mtc,
131i and
201the compound of the radioisotope labeling of the maximum element of content in the biological tissues such as Tl, they have the feature of with molecule in body similar (comprising cellular metabolism).And radionuclide
131i can decay and can launch gamma-rays and β ray.Gamma-rays can be used for biodistribution research in biological in-vivo imaging and body thereof, and β ray can be used to carry out radiation treatment.Therefore,
131i can be used as functional nucleic and carry out single photon emission computerized tomography (SPECT) and radiation treatment simultaneously.But due to molecule
131the blood circulation time that I is shorter in vivo and poor distribution of specific limit it and carry out biomedical applications as effective diagnosis and treatment nucleic.Therefore an effective load is built
131the nanosystems of I becomes the key of SPECT imaging and radiation treatment in body.
Daiamid (poly (amidoamine), PAMAM) dendrimer is by the branching unit progressively macromole with tree-shaped highly branched structure that obtains of reaction repeated.Molecular structure comprises three parts: multi-arm initiated core; The support be made up of multiple repetitive; Surface multi-functional group.It has highly branched, high degree of monodispersity, has structure that is controlled, rule and composition.It not only has unique surface physics, chemical property because surface has numerous functional groups, and has unique internal cavity structures.Surface has one deck functional group, can pass through chain-interionic interaction, the stable metal ions such as Acid-Base interaction, forms composite.(Zhang, the Y.Q. such as such as Shen Y.M.; Shen, Y.M et al.Journalof Medicinal Chemistry.2010,53,3262) the 5th generation PAMAM dendrimer utilizing end group to be amino for template modified metal ion chelating agent diethyl pentetic acid (DTPA) with chelating radioelement technetium (
99mand modify targeting agent folic acid and Polyethylene Glycol for SPECT imaging in body Tc).(Chen, the Q. such as Shi X.Y.; Zhang, G.X.; Shi, X.Y.et al.Biomaterials.2013,34,5200) the 5th generation PAMAM dendrimer utilizing end group to be amino is for template load gadolinium (Gd) ion and in-situ reducing prepares gold nano grain, form MR/CT bimodal imaging system, and modify targeted molecular etc. in dendrimer periphery, the nano material prepared has good biocompatibility, can carry out targeting and imaging effectively to cancer cell.Dendrimer, due to the physico-chemical property of its uniqueness, can be used as the carrier of multifunctional targeted molecular imaging probe, is also the excellent carrier of medicine.Therefore, physicochemical property that can be excellent according to dendrimer, utilize targeting group Imaging probe to be taken to the early diagnosis that target site realizes tumor, and realized the radiation treatment of cancer by the character of radionuclide uniqueness, the growth of effective anticancer.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of radionuclide
131the multi-functional tree-shaped macromolecular preparation method of the folate-targeted of I labelling, the inventive method preparation process is simple, and experiment condition is normal temperature and pressure, is easy to operation, the preparation procedure adopted can be used for the macromolecular preparation of other functional dendritic, has good use value.
A kind of radionuclide of the present invention
131the multi-functional tree-shaped macromolecular preparation method of the folate-targeted of I labelling, comprising:
(1) folic acid FA is dissolved in solvent, adds EDC and activate 2-3h, then add NH
2-PEG-COOH solution, stirring reaction 1-2d, obtains FA-PEG-COOH;
(2) by the 5th PAMAM dendrimer solution G5.NH
2add 3-(4-hydroxy phenyl) propanoic acid N-hydroxy-succinamide ester HPAO solution, stirring reaction 24h, obtains G5.NH
2-HPAO;
(3) aqueous solution of FA-PEG-COOH is activated 2-3h through EDC, then join G5.NH
2in the aqueous solution of-HPAO, stirring reaction 2-3d, obtains dendrimer G5.NH
2-HPAO-(PEG-FA) solution;
(4) by above-mentioned dendrimer G5.NH
2triethylamine N (C is added in-HPAO-(PEG-FA) solution
2h
5)
3stir 20-40min, finally add acetic anhydride Ac
2o, stirring reaction 12-24h, dialysis, lyophilization, obtains the dendrimer G5.NHAc-HPAO-(PEG-FA) of functionalization;
(5) toluene-sodium-sulfonchloramide (ch-T) and sodium iodide Na is added by the PBS solution of the dendrimer G5.NHAc-HPAO-(PEG-FA) of above-mentioned functions
131i, after reaction 2min, adds Na
2s
2o
5and potassium iodide, reaction 1min, separation and purification, obtains elemental radioiodine
131the nano material of I labelling
131i-G5.NHAc-HPAO-(PEG-FA).
In described step (1), (2), solvent is dimethyl sulfoxide DMSO.
In described step (1), the mol ratio of EDC and FA is 1:1, FA and NH
2the mol ratio of-PEG-COOH is 1.5-2:1.
In described step (2), the mol ratio of HPAO and the 5th PAMAM dendrimer is 12:1.
FA-PEG-COOH and G5.NH in described step (3)
2the mol ratio of-HPAO is the mol ratio of 50:1, EDC and FA-PEG-COOH is 20:1.
Triethylamine N (C in described step (4)
2h
5)
3, acetic anhydride Ac
2o and dendrimer G5.NH
2the mol ratio of-HPAO-(PEG-FA) is 120-660:100-550:1.
In described step (4), dialysis is for using cellulose dialysis film MWCO=14000, and dialyse 3-5 days in phosphate buffered solution and distilled water.
Na in described step (5)
131i radioactive activity is 185-370MBq.
In described step (5), separation and purification is: with PD-10 desalination chromatography column separating purification, with the PBS of pH=7.2-7.4 for mobile phase, every 15 are collected in same test tube, altogether collect 20 pipes, measure radioactive activity pure to collect with radioactivity instrument
131the functional dendritic macromole of I labelling.
Use PEG can increase the circulation time in vivo of this material in the present invention, extend SPECT contrast time and strengthen its radiotherapy effectiveness in vivo.
Targeted molecular FA is used to be modified at dendrimer surface to realize the targeting specific effect of its inside and outside in the present invention.
Acetylated to reduce its surface potential by dendrimer surface residual in the present invention, improves the biocompatibility of material.The alkaline environment of solution is kept to ensure carrying out smoothly of acetylization reaction with triethylamine.
In the present invention dendrimer G5.NHAc-HPAO-(PEG-FA) PBS solution in add Na
131i, and fully stirring makes
131the HPAO of I fully and on dendrimer G5.NHAc-HPAO-(PEG-FA) reacts, unreacted Na
131i is separated through PD-10 desalination chromatographic column, and collection obtains pure
131i-G5.NHAc-HPAO-(PEG-FA).
Use
1sPECT imaging in HNMR (hydrogen nuclear magnetic resonance), TLC (thin layer chromatography) test, MTT test, fluidic cell survey, the burnt ultramicroscope survey of copolymerization, body, tumor size change, mice with tumor body weight change, mice with tumor survival rate statistics characterize radionuclide
131the multi-functional tree-shaped macromolecular result of the folate-targeted of I labelling is as follows respectively:
(1)
1h NMR test result
1h NMR test result shows: prepare in the present invention
131in I-G5.NHAc-HPAO-(PEG-FA), a G5 has met 9.4 HPAO, 35 PEG, 20 FA, see Figure of description 2.
(2) TLC test result
TLC test result shows: the functional dendritic macromole prepared in the present invention
131i-G5.NHAc-HPAO-(PEG-FA) has good stability, under different time points (0h, 1h, 3h, 5h, 27h and 44h),
131the radio-chemical purity of I-G5.NHAc-HPAO-(PEG-FA), all more than 80%, does not have a large amount of
131i splits away off from carrier, see Figure of description 3.
(3) MTT test result
MTT test result shows: the dendrimer G5.NHAc-HPAO-(PEG-FA) of the function vector that the present invention prepares has good biocompatibility, after functional dendritic macromole hatching cancerous cell 24h, cell viability is still higher than 80%, its good biocompatibility can be used in later experiment in vivo, see Figure of description 4.
(4) cellular morphology result
Cellular morphology result shows: with the functional dendritic macromolecular carrier G5.NHAc-HPAO-(PEG-FA) (0.1 μM of variable concentrations, 1 μM, 5 μMs, 10 μMs and 20 μMs) hatching cancerous cell, cellular morphology is good, there is not obvious apoptosis sign, see Figure of description 5.Illustrate that the functional dendritic macromolecular carrier prepared in the present invention has good biocompatibility.
(5) fluidic cell test result
Fluidic cell test result shows: the functional dendritic macromolecular carrier G5.NHAc-FI-HPAO-PEG-FA through FI labelling prepared in the present invention is 1000nM and hatching cancerous cell 2h in concentration, to the cancerous cell of homofolic acid expression of receptor, there is targeting, illustrate that obtained material has specificity to homofolic acid receptor-expressing cells, see Figure of description 6 by the significant difference of fluorescence intensity.
(6) laser confocal microscope test
Laser confocal microscope test result shows: the functional dendritic macromolecular carrier G5.NHAc-FI-HPAO-PEG-FA through FI labelling prepared in the present invention is 1000nM and hatching cancerous cell 2h in concentration, to the cell of homofolic acid expression of receptor, there is targeting, can see that the functional dendritic macromole prepared enters in the Cytoplasm of homofolic acid receptor-expressing cells clearly by Laser Scanning Confocal Microscope picture, illustrate that obtained material has specificity to homofolic acid receptor-expressing cells, see Figure of description 7.This result is consistent with fluidic cell test result, the common targeting specific of material for cancerous cell that modification FA is described.
(7) SPECT imaging test in body
In body, SPECT imaging test result shows: prepare in the present invention
131the functional dendritic macromole of I labelling
131i-G5.NHAc-HPAO-(PEG-FA), by tail vein injection 200 μ L
131after I-G5.NHAc-HPAO-(PEG-FA) (18.5MBq) to mice with tumor, test SPECT imaging effect in the body of 2min, 30min, 1h, 2h, 3h, 4h, 6h, 16h, 24h.Result shows when 6h, 16h and 24h, and the tumor locus of mice with tumor has obvious SPECT imaging signal, and non-targeted material
131i-G5.NHAc-HPAO-mPEG do not have obvious SPECT imaging signal at the tumor locus of mice with tumor, illustrates to prepare
131i-G5.NHAc-HPAO-(PEG-FA) has good in-vivo tumour targeting SPECT imaging characteristic, see Figure of description 8a.And when tail vein injection 6h and 24h, dissect taking-up tumor locus and carry out SPECT imaging.Result shows when 6h, through targeting material
131the mice with tumor tumor locus ratio that I-G5.NHAc-HPAO-(PEG-FA) processes is through non-targeted material
131the mice with tumor tumor locus of I-G5.NHAc-HPAO-mPEG process has obviously strong SPECT imaging signal.And when 24h,
131the mice with tumor tumor locus that I-G5.NHAc-HPAO-(PEG-FA) processes has stronger SPECT imaging signal, but
131the mice with tumor tumor locus of I-G5.NHAc-HPAO-mPEG process has not had SPECT imaging signal, see Figure of description 8b.Illustrate through 24h,
131i-G5.NHAc-HPAO-mPEG removes from tumor locus, and metabolism goes out in body.Further illustrate
131i-G5.NHAc-HPAO-(PEG-FA) has good in-vivo tumour targeting SPECT imaging characteristic.
(8) mice with tumor tumor size change test
Mice with tumor tumor size change test result shows: prepare in the present invention
131the functional dendritic macromole of I labelling
131i-G5.NHAc-HPAO-(PEG-FA) has the effect of obvious Tumor suppression growth.Pass through
131the growth fraction of the mice with tumor tumor that I-G5.NHAc-HPAO-(PEG-FA) processes is through normal saline, Na
131i and
131the growth of the mice with tumor tumor of I-G5.NHAc-HPAO-mPEG process is slow, see Figure of description 9.
(9) mice with tumor body weight change test
Mice with tumor body weight change test result shows: prepare in the present invention
131the functional dendritic macromole of I labelling
131i-G5.NHAc-HPAO-(PEG-FA) does not have a significant effect the change of mice with tumor body weight.Warp
131the mice with tumor body weight change that I-G5.NHAc-HPAO-(PEG-FA) processes basic with through normal saline, Na
131i and
131the mice with tumor body weight change similar trend of I-G5.NHAc-HPAO-mPEG process, see Figure of description 10.
(10) mice with tumor survival rate statistics
The statistical result of mice with tumor survival rate shows: prepare in the present invention
131the functional dendritic macromole of I labelling
131i-G5.NHAc-HPAO-(PEG-FA) makes the mice with tumor death time postpone, through statistics
131the mice with tumor that I-G5.NHAc-HPAO-(PEG-FA) organizes is until 38d also has the mice with tumor of survival.And the mice with tumor of normal saline group is in heaven complete at 22d, Na
131the mice with tumor of I group is in heaven complete at 28d,
131the mice with tumor of I-G5.NHAc-HPAO-mPEG group is in heaven complete at 31d, see Figure of description 11.
Radionuclide prepared by the present invention
131the multi-functional dendrimer of the folate-targeted of I labelling, it has good stability and biocompatibility.In vitro cell experiment result prove functional dendritic macromolecular carrier G5.NHAc-HPAO-(PEG-FA) cancerous cell to homofolic acid expression of receptor there is targeting and
131the functional dendritic macromole of I labelling
131i-G5.NHAc-HPAO-(PEG-FA) has good stability, and has targeting SPECT imaging and targeting radiation treatment function in vivo.
Have a large amount of amino with surface, the Polyamidoamine Dendrimers that inside has a large amount of cavity and structure controllable precise is template and stabilizing agent, has prepared radionuclide
131the multi-functional dendrimer of the folate-targeted of I labelling, the present invention relates to two ultimate principles:
(1) the modified change amino that the surface of Polyamidoamine Dendrimers is a large amount of is made full use of, at its finishing targeting group folic acid (FA) and 3-(4-hydroxy phenyl) propanoic acid N-hydroxy-succinamide ester (HPAO), Multifunction dendrimer can be prepared.
(2) with NH
2-PEG-COOH is intermediary, by FA and NH
2-PEG-COOH chemical bonding, then the FA-PEG-COOH of generation is connected with Polyamidoamine Dendrimers, biocompatibility and the body inner blood circulation time of this nano material can be improved.
Prepare this radionuclide
131the multi-functional tree-shaped macromolecular key element of the folate-targeted of I labelling is exactly find a suitable carrier platform with the different functional group of load, forms multifunctional nano complex.The present invention utilizes ad hoc structure and the character of dendrimer, targeting group folic acid (FA) and 3-(4-hydroxy phenyl) propanoic acid N-hydroxy-succinamide ester (HPAO) is modified at dendrimer surface to realize targeting and labelling
131the function of I, wherein with NH
2-PEG-COOH is intermediary, by FA and NH
2-PEG-COOH chemical bonding; again the FA-PEG-COOH of generation is connected with Polyamidoamine Dendrimers; the macromolecular biocompatibility of this functional dendritic and blood circulation time can be improved; and acetylation dendrimer surface residual amino reduces positive surface charge; to reduce its toxicity; prepare multifunction dendrimer nano platform, finally load on this functional dendritic macromole
131i, is expected to for SPECT imaging and radiotherapy integration.
beneficial effect
(1) preparation process of the present invention is simple, and experiment condition is normal temperature and pressure, and be easy to operation, the preparation procedure adopted can be used for the macromolecular preparation of other functional dendritic, has good use value;
(2) material prepared by the present invention can be used for in-vivo tumour SPECT and diagnoses, and has good diagnostic application prospect;
(3) radionuclide for preparing of the present invention
131the multi-functional dendrimer of the folate-targeted of I labelling has good targeting radiotherapeutic efficacy, for the radiopharmaceutic exploitation of novel nano is laid a good foundation.
Accompanying drawing explanation
Fig. 1 is reaction equation sketch of the present invention;
Fig. 2 is (a) FA-PEG-COOH prepared by the present invention, (b) G5.NH
2the hydrogen nuclear magnetic resonance spectrogram of-HPAO, (c) G5.NHAc-HPAO-PEG-FA, (d) G5.NHAc-HPAO-mPEG;
Fig. 3 is Na
131prepared by I (a) and the present invention
131i-G5.NHAc-HPAO-PEG-FA is at 0h (b), 1h (c), 3h (d), 5h (e), the radio-chemical purity test of 27h (f), 44h (g) and radio chemistry Efficiency Statistics (h) thereof;
Fig. 4 is through variable concentrations
131i-G5.NHAc-HPAO-PEG-FA hatches the C6 cell MTT test result figure of 24h;
Fig. 5 is through PBS (a) and using 0.1 μM (b) respectively under phase contrast microscope, 1 μM (c), 5 μMs (d), 10 μMs (e), the G5.NHAc-HPAO-PEG-FA of 20 μMs (f) hatches the cellular morphology figure after C6 cell 24h;
Fig. 6 is (a) C6-HFAR cell PBS process; (b), the G5.NHAc-FI-HPAO-mPEG of (c) C6-LFAR cell and C6-HFAR cell 1000nM processes 2h respectively; (d), the G5.NHAc-FI-HPAO-PEG-FA of (e) C6-LFAR cell and C6-HFAR cell 1000nM processes the flow cytometry figure of 2h respectively; F Fluorescence Intensity Assays datagram that () is C6 cell; C6-HFAR refers to the cell of homofolic acid expression of receptor, and C6-LFAR refers to the cell that low folacin receptor is expressed;
Fig. 7 is respectively with the different materials hatching C6-LFAR cell of 2h and the Laser Scanning Confocal Microscope figure of C6-HFAR cell that concentration is 1000nM;
Fig. 8 is mice with tumor tail vein injection
131i-G5.NHAc-HPAO-PEG-FA and
131sPECT image (a) of mice with tumor and SPECT imaging (b) at 6h and 24h tumor locus after dissecting under different time points after I-G5.NHAc-HPAO-mPEG;
Fig. 9 is the gross tumor volume variation diagram of mice with tumor after different materials process;
Figure 10 is the body weight change trendgram of mice with tumor after different materials process;
Figure 11 is the survival rate statistical analysis figure of mice with tumor after different materials process.
Detailed description of the invention
Below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention.In addition should be understood that those skilled in the art can make various changes or modifications the present invention, and these equivalent form of values fall within the application's appended claims limited range equally after the content of having read the present invention's instruction.
Embodiment 1
(1) dissolving dry weight with the DMSO of 5mL is the FA of 11.37mg, dropwise dripping the dry weight being dissolved in 2mL DMSO solution is while stirring the EDC of 4.94mg, after stirring reaction 3h, in this solution, dropwise drip the dry weight being dissolved in 5mL DMSO solution is the NH of 85.86mg
2-PEG-COOH (Mw=5000), wherein FA and NH
2the mol ratio of-PEG-COOH is the mol ratio of 1.5:1, EDC and FA is 1:1, and react 1 day, dialysed 3 days in phosphate buffered solution and distilled water by the solution of gained cellulose dialysis film MWCO=1000, lyophilization process obtains FA-PEG-COOH.
(2) dissolve with the DMSO of 5mL the G5PAMAM dendrimer that dry weight is 30mg, dropwise dripping the dry weight being dissolved in 2mL DMSO solution is while stirring the HPAO of 3.64mg, wherein the mol ratio of HPAO and G5PAMAM dendrimer is 12:1, react 1 day, dialysed 3 days in phosphate buffered solution and distilled water by the solution of gained cellulose dialysis film MWCO=14000, lyophilization process obtains G5.NH
2-HPAO.
(3) dissolving dry weight with the distilled water of 5mL is the FA-PEG-COOH of 30mg, dropwise dripping the dry weight being dissolved in 2mL distilled water solution is while stirring the EDC of 21.9mg, after stirring reaction 3h, in this solution, dropwise drip the dry weight being dissolved in 5mL distilled water is the G5.NH of 3.27mg
2-HPAO dendrimer, wherein the mol ratio of FA-PEG-COOH and G5PAMAM dendrimer is 50:1, the mol ratio of EDC and FA-PEG-COOH is 20:1, react 3 days, dialysed 3 days in phosphate buffered solution and distilled water by the solution of gained cellulose dialysis film MWCO=14000, lyophilization process obtains G5.NH
2-HPAO-(PEG-FA).
(4) dissolving dry weight with the distilled water of 5mL is the G5.NH of 15mg
2-HPAO-(PEG-FA), dropwise drip 6.55 μ L triethylamines while stirring, after stirring reaction 30min, 3.70 μ L acetic anhydrides are added in reactant liquor, at room temperature stirring reaction 24 hours, dialysed 3 days in phosphate buffered solution and distilled water by the solution of gained cellulose dialysis film MWCO=14000, lyophilization process obtains multifunction dendrimer nano platform G5.NHAc-HPAO-PEG-FA.
In building-up process, dendrimer finishing nuclear-magnetism is characterized, integral and calculating carried out to each peak known: dendrimer supports finishing 9.4 HPAO, 35 PEG, 20 FA molecules (accompanying drawing 2).These test results show the dendrimer G5.NHAc-HPAO-PEG-FA of successful design complex functionality.And at labelling
131after I, test is formed
131i-G5.NHAc-HPAO-PEG-FA radio-chemical purity reaches 97.7% ± 0.7%, and labelling is described
131i success.
Embodiment 2
With thin layer chromatography to the product obtained
131i-G5.NHAc-HPAO-PEG-FA carries out stability analysis, see accompanying drawing 3.The radioactive marker of 100 μ L is mixed, at utilizing thin layer chromatography to test 37 DEG C afterwards with the normal saline of 1mL 0.9% and hyclone respectively
131the radio-chemical purity of I-G5.NHAc-HPAO-PEG-FA when 0h, 1h, 3h, 5h, 27h and 44h, result shows under different time points,
131the radio-chemical purity of I-G5.NHAc-HPAO-PEG-FA, all more than 80%, does not have a large amount of
131i splits away off from carrier.The product obtained is described
131i-G5.NHAc-HPAO-PEG-FA has good stability.
Embodiment 3
The biocompatibility in vitro of the multi-functional dendrimer supports of the folate-targeted prepared is detected with MTT test, take carrier material 12.7mg in embodiment 1, be dissolved in the PBS buffer of 300 μ L and prepare the solution that G5.NHAc-HPAO-PEG-FA concentration is 200 μMs, being diluted to concentration is more respectively 100 μMs, 50 μMs, 10 μMs, the solution of 1 μM.By the material of above-mentioned variable concentrations, (its respective final concentration is 20 μMs, 10 μMs, 5 μMs, 1 μM, 0.1 μM) after hatching C6 cell 24h, material is outwelled, add 180 μ L culture medium and 20 μ L MTT, after 37 DEG C of hatching 4h, culture fluid is outwelled, adds 200 μ L DMSO, after shaking table jolting 20min, record result by microplate reader, obtain data shown in Fig. 4, result shows that embodiment 1 is unmarked within the scope of finite concentration
131the carrier material of I does not have obvious cytotoxicity, has good biocompatibility in vitro.
Embodiment 4
The biocompatibility of prepared carrier material is analyzed further by cellular morphology, take 16.98mgG5.NHAc-HPAO-PEG-FA, be dissolved in the PBS of 400 μ L and prepare the solution that G5.NHAc-HPAO-PEG-FA concentration is 200 μMs, being diluted to concentration is more respectively 100 μMs, 50 μMs, 10 μMs, the solution of 1 μM.By C6 cell kind in 96 orifice plates, 10
4individual cells/well, after hatching 24h, adds 10 μ LPBS in every hole respectively, the material of above-mentioned variable concentrations and 90 μ L culture medium, make the final concentration of each material be 20 μMs, 10 μMs, 5 μMs, 1 μM, 0.1 μM, after hatching C6 cell 24h, outwelled by material, use phase contrast microscope shooting cell image, result shows that the cellular morphology of hatching with PBS and each concentration G5.NHAc-HPAO-PEG-FA is good, there is no obvious apoptosis sign, see accompanying drawing 5.Illustrate in the present invention that the multi-functional dendrimer supports preparing folate-targeted does not have cytotoxicity, there is good cell compatibility.
Embodiment 5
The targeting of the multi-functional dendrimer supports material of the folate-targeted prepared is proved with fluidic cell test.By carrier material mark fluorescent tracer Fluorescein isothiocyanate (FI) in embodiment 1, then take the embodiment 1 carrier material 2.14mg through FI labelling, being dissolved in compound concentration in the PBS buffer of 1000 μ L is the solution of 10000nM; Meanwhile also by the carrier material flag F I in comparative example 1, take the comparative example 1 material 2.05mg through FI labelling, being dissolved in compound concentration in the PBS buffer of 1000 μ L is the solution of 10000nM; By C6 cell kind in 12 orifice plates, 2 × 10
5cells/well, C6 cell is divided into homofolic acid expression of receptor and low folacin receptor to express, and uses 2.5 μMs of acid-treated cells of leaf to be the C6 cell that low folacin receptor is expressed, does not use the C6 cell that the acid-treated cell of leaf is homofolic acid expression of receptor.Each cell uses isopyknic PBS process, after hatching 24h, embodiment 1 carrier material of 100 μ L through FI labelling is added respectively in every hole, through comparative example 1 material and the 900 μ L culture medium of FI labelling, the final concentration of each material is 1000nM, by material sucking-off after hatching 2h, wash 2-3 time with PBS, with trypsin by cell suspension, after centrifugal, be dissolved in 1mLPBS, flow cytometer is used to measure, result shows that the G5.NHAc-FI-HPAO-PEG-FA with concentration is 1000nM hatches cancerous cell 2h, to the cancerous cell of homofolic acid expression of receptor, there is targeting, the comparative example 1 material G5.NHAc-FI-HPAO-mPEG through FI labelling not with folic acid does not have a targeting to the cancerous cell that height folacin receptor is expressed, illustrate that obtained embodiment 1 carrier material through FI labelling has specificity to homofolic acid receptor-expressing cells by the significant difference of fluorescence intensity, see Figure of description 6.
Embodiment 6
The targeting of the multi-functional dendrimer supports material of the folate-targeted prepared is proved further with Laser Scanning Confocal Microscope test.Take the embodiment 1 carrier material 2.14mg through FI labelling, being dissolved in compound concentration in the PBS buffer of 1000 μ L is the solution of 10000nM; Take the comparative example 1 material 2.05mg through FI labelling, being dissolved in compound concentration in the PBS buffer of 1000 μ L is the solution of 10000nM; By C6 cell kind in 12 orifice plates being placed with coverslip, 8 × 10
4cells/well, C6 cell is divided into homofolic acid expression of receptor and low folacin receptor to express, and uses 2.5 μMs of acid-treated cells of leaf to be the C6 cell that low folacin receptor is expressed, does not use the C6 cell that the acid-treated cell of leaf is homofolic acid expression of receptor.Each cell uses isopyknic PBS process, after hatching 24h, embodiment 1 carrier material of 100 μ L through FI labelling is added respectively in every hole, through comparative example 1 material and the 900 μ L culture medium of FI labelling, the final concentration of each material is 1000nM, by material sucking-off after hatching 2h, wash 2-3 time with PBS, every hole adds 2.5% glutaraldehyde 500 μ L and fixes 15min, wash 2-3 time with PBS again, after adding 1 μ g/mL Hoechst33342500 μ L dyeing 20min, wash 2-3 time with PBS, by the cell mounting on coverslip on microscope slide, take under Laser Scanning Confocal Microscope, result shows that the G5.NHAc-FI-HPAO-PEG-FA with concentration is 1000nM hatches cancerous cell 2h, to the cancerous cell of homofolic acid expression of receptor, there is targeting, the comparative example 1 material G5.NHAc-FI-HPAO-mPEG through FI labelling not with folic acid does not have a targeting to the cancerous cell that height folacin receptor is expressed, can see that embodiment 1 carrier material through FI labelling enters in the Cytoplasm of homofolic acid receptor-expressing cells clearly by Laser Scanning Confocal Microscope picture, illustrate that obtained carrier material has specificity to homofolic acid receptor-expressing cells, see accompanying drawing 7.This result is consistent with fluidic cell test result, common illustrates that embodiment 1 carrier material through FI labelling is for the targeting specific of cancerous cell.
Embodiment 7
The radionuclide obtained is tested with SPECT imager
131the multi-functional dendrimer of the folate-targeted of I labelling
131the SPECT imaging effect of I-G5.NHAc-HPAO-PEG-FA.Using the male nude mouse of about 20g as animal model (Shanghai animal experimental center), inoculated tumour, treats that tumor grows to 1.0cm
3after, prepared by tail vein injection 200 μ L the present invention
131i-G5.NHAc-HPAO-PEG-FA (18.5MBq) enters in mice with tumor body, carries out SPECT scanning (accompanying drawing 8a) in different time points (2min, 30min, 1h, 2h, 3h, 4h, 6h, 16h, 24h).Find out from picture 8a, compared with matched group (the mice with tumor picture of non-folate targeting material processed), the SPECT signal intensity of the mice with tumor tumor locus scanned after injection 6h, 16h, 24h obviously strengthens, and shows
131i-G5.NHAc-HPAO-PEG-FA material has target tumor SPECT imaging effect.Then when 6h and 24h, mice with tumor is dissected, take out tumor and carry out SPECT imaging (accompanying drawing 8b), finding out, during 6h from picture 8b
131the SPECT signal of I-G5.NHAc-HPAO-PEG-FA obviously than
131the SPECT signal of I-G5.NHAc-HPAO-mPEG is strong.When 24h,
131the SPECT signal of I-G5.NHAc-HPAO-PEG-FA is stronger than its SPECT signal at 6h, and when 24h
131the SPECT weak output signal of I-G5.NHAc-HPAO-mPEG, illustrates contrast material
131i-G5.NHAc-HPAO-mPEG has been fallen by slowly metabolism when 24h.By under different time points
131i-G5.NHAc-HPAO-PEG-FA with
131the SPECT signal value contrast of I-G5.NHAc-HPAO-mPEG demonstrates
131i-G5.NHAc-HPAO-PEG-FA is to the targeting SPECT imaging effect of tumor.
Embodiment 8
Described in embodiment 7, set up tumor model, treat that tumor grows to 0.5-1.2cm
3after, every 3 days tail vein injection salines (200 μ L, 7.4MBq), Na
131i (200 μ L, 7.4MBq),
131i-G5.NHAc-HPAO-mPEG (200 μ L, 7.4MBq) and
131i-G5.NHAc-HPAO-PEG-FA (200 μ L, 7.4MBq).Tumor size was measured every 3 days with slide gauge, and according to (tumor major diameter × (tumor minor axis)
2)/2 calculate tumor size, the relative tumour volume (V/V of mice with tumor
0, V is each tumor size measured, V
0be the tumor size of the 0th day) (accompanying drawing 9) and every 3 days of body weight change (accompanying drawing 10) record once, meanwhile record the death condition of mice with tumor, do fatality rate statistics (accompanying drawing 11).From accompanying drawing 9, through the mice with tumor tumor growth rate of different materials process
131i-G5.NHAc-HPAO-PEG-FA <
131i-G5.NHAc-HPAO-mPEG < Na
131i < normal saline.With
131i-G5.NHAc-HPAO-mPEG compares,
131i-G5.NHAc-HPAO-PEG-FA has targeting radiotherapeutic efficacy, and its interior therapeutic efficiency is than pure Na
131i is obvious.From accompanying drawing 10, warp
131after I-G5.NHAc-HPAO-PEG-FA process, obviously do not affect mice with tumor body weight change, warp
131the mice with tumor body weight change that I-G5.NHAc-HPAO-(PEG-FA) processes basic with through normal saline, Na
131i and
131the mice with tumor body weight change similar trend of I-G5.NHAc-HPAO-mPEG process, illustrates that it has biocompatibility in good body.From accompanying drawing 11,
131i-G5.NHAc-HPAO-(PEG-FA) makes the mice with tumor death time postpone, through statistics
131the mice with tumor that I-G5.NHAc-HPAO-(PEG-FA) organizes is until 38d also has the mice with tumor of survival.And the mice with tumor of normal saline group is in heaven complete at 22d, Na
131the mice with tumor of I group is in heaven complete at 28d,
131the mice with tumor of I-G5.NHAc-HPAO-mPEG group is in heaven complete at 31d, explanation
131i-G5.NHAc-HPAO-(PEG-FA) has good therapeutic efficiency, can effectively delay the mice with tumor death time.
Comparative example 1
(1) dissolve with the DMSO of 5mL the G5PAMAM dendrimer that dry weight is 30mg, dropwise dripping the dry weight being dissolved in 2mL DMSO solution is while stirring the HPAO of 3.64mg, wherein the mol ratio of HPAO and G5PAMAM dendrimer is 12:1, reaction 1d, dialysed 3 days in phosphate buffered solution and distilled water by the solution of gained cellulose dialysis film MWCO=14000, lyophilization process obtains G5.NH
2-HPAO.
(2) dissolve with the distilled water of 5mL the mPEG-COOH (Mw=5000) that dry weight is 30mg, dropwise dripping the dry weight being dissolved in 2mL distilled water solution is while stirring the EDC of 23mg, after stirring reaction 3h, in this solution, dropwise drip the dry weight being dissolved in 5mL distilled water is the G5.NH of 3.42mg
2-HPAO dendrimer, wherein the mol ratio of mPEG-COOH and G5PAMAM dendrimer is 50:1, the mol ratio of EDC and mPEG-COOH is 20:1, react 3 days, dialysed 3 days in phosphate buffered solution and distilled water by the solution of gained cellulose dialysis film MWCO=14000, lyophilization process obtains G5.NH
2-HPAO-mPEG.
(3) dissolving dry weight with the distilled water of 5mL is the G5.NH of 20mg
2-HPAO-mPEG, dropwise drip 9.12 μ L triethylamines while stirring, after stirring reaction 30min, 5.16 μ L acetic anhydrides are added in reactant liquor, at room temperature stirring reaction 24 hours, dialysed 3 days in phosphate buffered solution and distilled water by the solution of gained cellulose dialysis film MWCO=14000, lyophilization process obtains multifunction dendrimer nano platform G5.NHAc-HPAO-mPEG.
In building-up process, this comparison vehicle material nuclear-magnetism is characterized, integral and calculating carried out to each peak known: dendrimer supports finishing 9.4 HPAO, 35 PEG.These test results show successful design synthesis comparison vehicle material G5.NHAc-HPAO-mPEG.And at labelling
131after I, test is formed
131i-G5.NHAc-HPAO-mPEG radio-chemical purity, more than 98%, illustrates labelling
131i success.
Claims (9)
1. a radionuclide
131the multi-functional tree-shaped macromolecular preparation method of the folate-targeted of I labelling, comprising:
(1) folic acid FA is dissolved in solvent, adds EDC and activate 2-3h, then add NH
2-PEG-COOH solution, stirring reaction 1-2d, obtains FA-PEG-COOH;
(2) by the 5th PAMAM dendrimer solution G5.NH
2add 3-(4-hydroxy phenyl) propanoic acid N-hydroxy-succinamide ester HPAO solution, stirring reaction 24h, obtains G5.NH
2-HPAO;
(3) aqueous solution of FA-PEG-COOH is activated 2-3h through EDC, then join G5.NH
2in the aqueous solution of-HPAO, stirring reaction 2-3d, obtains dendrimer G5.NH
2-HPAO-(PEG-FA) solution;
(4) by above-mentioned dendrimer G5.NH
2triethylamine N (C is added in-HPAO-(PEG-FA) solution
2h
5)
3stir 20-40min, finally add acetic anhydride Ac
2o, stirring reaction 12-24h, dialysis, lyophilization, obtains the dendrimer G5.NHAc-HPAO-(PEG-FA) of functionalization;
(5) toluene-sodium-sulfonchloramide (ch-T) and sodium iodide Na is added by the PBS solution of the dendrimer G5.NHAc-HPAO-(PEG-FA) of above-mentioned functions
131i, after reaction 2min, adds Na
2s
2o
5and potassium iodide, reaction 1min, separation and purification, obtains elemental radioiodine
131the nano material of I labelling
131i-G5.NHAc-HPAO-(PEG-FA).
2. a kind of radionuclide according to claim 1
131the multi-functional tree-shaped macromolecular preparation method of the folate-targeted of I labelling, is characterized in that: in described step (1), (2), solvent is dimethyl sulfoxide DMSO.
3. a kind of radionuclide according to claim 1
131the multi-functional tree-shaped macromolecular preparation method of the folate-targeted of I labelling, is characterized in that: in described step (1), the mol ratio of EDC and FA is 1:1, FA and NH
2the mol ratio of-PEG-COOH is 1.5-2:1.
4. a kind of radionuclide according to claim 1
131the multi-functional tree-shaped macromolecular preparation method of the folate-targeted of I labelling, is characterized in that: in described step (2), the mol ratio of HPAO and the 5th PAMAM dendrimer is 12:1.
5. a kind of radionuclide according to claim 1
131the multi-functional tree-shaped macromolecular preparation method of the folate-targeted of I labelling, is characterized in that: FA-PEG-COOH and G5.NH in described step (3)
2the mol ratio of-HPAO is the mol ratio of 50:1, EDC and FA-PEG-COOH is 20:1.
6. a kind of radionuclide according to claim 1
131the multi-functional tree-shaped macromolecular preparation method of the folate-targeted of I labelling, is characterized in that: triethylamine N (C in described step (4)
2h
5)
3, acetic anhydride Ac
2o and dendrimer G5.NH
2the mol ratio of-HPAO-(PEG-FA) is 120-660:100-550:1.
7. a kind of radionuclide according to claim 1
131the multi-functional tree-shaped macromolecular preparation method of the folate-targeted of I labelling, is characterized in that: in described step (4), dialysis is for using cellulose dialysis film MWCO=14000, and dialyse 3-5 days in phosphate buffered solution and distilled water.
8. a kind of radionuclide according to claim 1
131the multi-functional tree-shaped macromolecular preparation method of the folate-targeted of I labelling, is characterized in that: Na in described step (5)
131i radioactive activity is 185-370MBq.
9. a kind of radionuclide according to claim 1
131the multi-functional tree-shaped macromolecular preparation method of the folate-targeted of I labelling, it is characterized in that: in described step (5), separation and purification is: with PD-10 desalination chromatography column separating purification, with the PBS of pH=7.2-7.4 for mobile phase, every 15 are collected in same test tube, altogether collect 20 pipes, measure radioactive activity with radioactivity instrument pure to collect
131the functional dendritic macromole of I labelling.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510171048.3A CN104758959B (en) | 2015-04-10 | 2015-04-10 | A kind of radionuclide131The preparation method of the multi-functional dendrimer of the folate-targeted of I marks |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510171048.3A CN104758959B (en) | 2015-04-10 | 2015-04-10 | A kind of radionuclide131The preparation method of the multi-functional dendrimer of the folate-targeted of I marks |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104758959A true CN104758959A (en) | 2015-07-08 |
| CN104758959B CN104758959B (en) | 2018-01-05 |
Family
ID=53641267
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510171048.3A Expired - Fee Related CN104758959B (en) | 2015-04-10 | 2015-04-10 | A kind of radionuclide131The preparation method of the multi-functional dendrimer of the folate-targeted of I marks |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104758959B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106948810A (en) * | 2017-04-10 | 2017-07-14 | 河南省科学院同位素研究所有限责任公司 | A kind of preparation method of hydrophobic liquid radioactive tracer |
| CN109865145A (en) * | 2019-03-12 | 2019-06-11 | 东华大学 | A kind of radionuclide131The preparation method of the functional poly phosphonitrile nanosphere of I label |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101294955A (en) * | 2008-06-06 | 2008-10-29 | 吉林大学 | Production method of probe molecule for recognizing tumor cell by specificity |
| CN101327328A (en) * | 2008-07-29 | 2008-12-24 | 华东师范大学 | Dendrimers targeting nano particle and preparation and application thereof |
| CN101531800A (en) * | 2009-04-14 | 2009-09-16 | 东华大学 | Method for preparing poly(amidoamine)/carbon nanometer tube composite material for cancer cell targeting diagnosis |
| CN101569751A (en) * | 2009-06-09 | 2009-11-04 | 华东师范大学 | Tumor-targeted nonionic dendritic macromolecule magnetic resonance imaging contrast agent |
| CN104162175A (en) * | 2014-06-18 | 2014-11-26 | 东华大学 | Functionalized dendrimer-based SPECT-CT bimodal imaging contrast agent and preparation method thereof |
| US20140371401A1 (en) * | 2013-06-17 | 2014-12-18 | Universidad De Talca | Pamam, spacer molecule and cafestol polymers |
-
2015
- 2015-04-10 CN CN201510171048.3A patent/CN104758959B/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101294955A (en) * | 2008-06-06 | 2008-10-29 | 吉林大学 | Production method of probe molecule for recognizing tumor cell by specificity |
| CN101327328A (en) * | 2008-07-29 | 2008-12-24 | 华东师范大学 | Dendrimers targeting nano particle and preparation and application thereof |
| CN101531800A (en) * | 2009-04-14 | 2009-09-16 | 东华大学 | Method for preparing poly(amidoamine)/carbon nanometer tube composite material for cancer cell targeting diagnosis |
| CN101569751A (en) * | 2009-06-09 | 2009-11-04 | 华东师范大学 | Tumor-targeted nonionic dendritic macromolecule magnetic resonance imaging contrast agent |
| US20140371401A1 (en) * | 2013-06-17 | 2014-12-18 | Universidad De Talca | Pamam, spacer molecule and cafestol polymers |
| CN104162175A (en) * | 2014-06-18 | 2014-11-26 | 东华大学 | Functionalized dendrimer-based SPECT-CT bimodal imaging contrast agent and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| ZHANG Y, ET AL.: "Synthesis, biodistribution, and microsingle photon emission computed tomography(SPECT) imaging study of technetium-99m labeled PEGylated dendrimer poly(amidoamine)(PAMAM)-folic acid conjugates", 《J MED CHEM》 * |
| 董文宾: "《生物工程分析》", 30 April 2006 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106948810A (en) * | 2017-04-10 | 2017-07-14 | 河南省科学院同位素研究所有限责任公司 | A kind of preparation method of hydrophobic liquid radioactive tracer |
| CN109865145A (en) * | 2019-03-12 | 2019-06-11 | 东华大学 | A kind of radionuclide131The preparation method of the functional poly phosphonitrile nanosphere of I label |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104758959B (en) | 2018-01-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chen et al. | Fabrication of fluorescent nanoparticles based on AIE luminogens (AIE dots) and their applications in bioimaging | |
| Qiao et al. | Dendrimer-based molecular imaging contrast agents | |
| Cao et al. | Biodistribution of sub-10 nm PEG-modified radioactive/upconversion nanoparticles | |
| Ma et al. | 64Cu-Labeled multifunctional dendrimers for targeted tumor PET imaging | |
| Chen et al. | Dual-modal MRI contrast agent with aggregation-induced emission characteristic for liver specific imaging with long circulation lifetime | |
| Zhou et al. | Tumor-homing peptide-based NIR-II probes for targeted spontaneous breast tumor imaging | |
| Kim et al. | Gadolinium-chelate nanoparticle entrapped human mesenchymal stem cell via photochemical internalization for cancer diagnosis | |
| Zhang et al. | Affibody-functionalized Ag 2 S quantum dots for photoacoustic imaging of epidermal growth factor receptor overexpressed tumors | |
| CN104436220B (en) | A kind of preparation method and its usage of chitosan magnetic Nano microsphere | |
| CN104350109A (en) | Functionalised silicon nanoparticles | |
| Fernández-Barahona et al. | Molecular imaging with 68Ga radio-nanomaterials: shedding light on nanoparticles | |
| CN110128666A (en) | Functionalized polyethy-lene imines wraps up nanogold particle composite material and preparation method | |
| CN106075469A (en) | Gd3+induction gold nano cluster is self-assembled into method and the application of gold nano grain | |
| Jiménez Sánchez et al. | Fluorescent radiosensitizing gold nanoparticles | |
| CN105079826A (en) | Preparation method and application of RGD@BBN double-targeted MR (magnetic resonance)/optical dual-mode molecular probe | |
| CN104162175B (en) | Functionalized dendrimer-based SPECT-CT bimodal imaging contrast agent and preparation method thereof | |
| Boschi et al. | Nanoparticles for Cerenkov and radioluminescent light enhancement for imaging and radiotherapy | |
| Jarockyte et al. | Biodistribution of multimodal gold nanoclusters designed for photoluminescence-SPECT/CT imaging and diagnostic | |
| CN105770922B (en) | It is a kind of64The preparation method for the trimanganese tetroxide nano particle that Cu marks the functionalized polyethy-lene imines of folate-targeted stable | |
| CN104758959A (en) | Preparation method of radionuclide 131I-labeled folic acid targeted multifunctional dendrimer | |
| Kubeil et al. | Dual-labelling strategies for nuclear and fluorescence molecular imaging: current status and future perspectives | |
| CN107118767B (en) | Radionuclide iodine labeled fluorescent carbon dot, synthesis method and application | |
| CN102070747A (en) | Polymer for fluorescein angiogram and isotope angiogram | |
| Zhu et al. | Charge-conversional polyethylenimine-entrapped gold nanoparticles with 131 I-labeling for enhanced dual mode SPECT/CT imaging and radiotherapy of tumors | |
| CN104258420A (en) | Preparation method of CT nano contrast agent namely low-algebraic tree-shaped macromolecule-coated gold nano particles with liver cancer targeting function |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180105 Termination date: 20200410 |