CN102690328B - Single photon emission computed tomography developer and preparation method thereof - Google Patents
Single photon emission computed tomography developer and preparation method thereof Download PDFInfo
- Publication number
- CN102690328B CN102690328B CN201210164547.6A CN201210164547A CN102690328B CN 102690328 B CN102690328 B CN 102690328B CN 201210164547 A CN201210164547 A CN 201210164547A CN 102690328 B CN102690328 B CN 102690328B
- Authority
- CN
- China
- Prior art keywords
- compounds
- reaction
- structural formula
- add
- mmole
- 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.)
- Expired - Fee Related
Links
- 238000002360 preparation method Methods 0.000 title claims abstract description 38
- 238000002603 single-photon emission computed tomography Methods 0.000 title abstract description 3
- 150000001875 compounds Chemical class 0.000 claims abstract description 283
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 32
- 238000000034 method Methods 0.000 claims abstract description 19
- 238000010532 solid phase synthesis reaction Methods 0.000 claims abstract description 6
- 238000006243 chemical reaction Methods 0.000 claims description 69
- 150000001413 amino acids Chemical class 0.000 claims description 52
- 235000001014 amino acid Nutrition 0.000 claims description 50
- 125000006239 protecting group Chemical group 0.000 claims description 38
- 239000004475 Arginine Substances 0.000 claims description 36
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 30
- 230000004913 activation Effects 0.000 claims description 30
- 238000012360 testing method Methods 0.000 claims description 26
- 238000003325 tomography Methods 0.000 claims description 26
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 24
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 24
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 23
- 239000000203 mixture Substances 0.000 claims description 23
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 21
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 20
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 claims description 18
- 239000000047 product Substances 0.000 claims description 15
- VDQQXEISLMTGAB-UHFFFAOYSA-N chloramine T Chemical compound [Na+].CC1=CC=C(S(=O)(=O)[N-]Cl)C=C1 VDQQXEISLMTGAB-UHFFFAOYSA-N 0.000 claims description 14
- 229960001479 tosylchloramide sodium Drugs 0.000 claims description 14
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims description 14
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims description 13
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 claims description 12
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 claims description 12
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 12
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 12
- IOJUPLGTWVMSFF-UHFFFAOYSA-N benzothiazole Chemical compound C1=CC=C2SC=NC2=C1 IOJUPLGTWVMSFF-UHFFFAOYSA-N 0.000 claims description 12
- 239000011347 resin Substances 0.000 claims description 12
- 229920005989 resin Polymers 0.000 claims description 12
- 239000002904 solvent Substances 0.000 claims description 12
- 238000003756 stirring Methods 0.000 claims description 12
- HRZFUMHJMZEROT-UHFFFAOYSA-L sodium disulfite Chemical compound [Na+].[Na+].[O-]S(=O)S([O-])(=O)=O HRZFUMHJMZEROT-UHFFFAOYSA-L 0.000 claims description 11
- 235000010262 sodium metabisulphite Nutrition 0.000 claims description 11
- 239000004296 sodium metabisulphite Substances 0.000 claims description 11
- 238000002347 injection Methods 0.000 claims description 9
- 239000007924 injection Substances 0.000 claims description 9
- 238000000926 separation method Methods 0.000 claims description 9
- 239000002250 absorbent Substances 0.000 claims description 8
- 230000002745 absorbent Effects 0.000 claims description 8
- -1 9-fluorenylmethyloxycarbonyl Chemical group 0.000 claims description 7
- 239000003153 chemical reaction reagent Substances 0.000 claims description 7
- 239000000243 solution Substances 0.000 claims description 7
- YOETUEMZNOLGDB-UHFFFAOYSA-N 2-methylpropyl carbonochloridate Chemical compound CC(C)COC(Cl)=O YOETUEMZNOLGDB-UHFFFAOYSA-N 0.000 claims description 6
- 239000000460 chlorine Substances 0.000 claims description 6
- 229910052801 chlorine Inorganic materials 0.000 claims description 6
- 229960004126 codeine Drugs 0.000 claims description 6
- OROGSEYTTFOCAN-DNJOTXNNSA-N codeine Chemical compound C([C@H]1[C@H](N(CC[C@@]112)C)C3)=C[C@H](O)[C@@H]1OC1=C2C3=CC=C1OC OROGSEYTTFOCAN-DNJOTXNNSA-N 0.000 claims description 6
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims description 6
- 235000018417 cysteine Nutrition 0.000 claims description 6
- XMBWDFGMSWQBCA-RNFDNDRNSA-M iodine-131(1-) Chemical compound [131I-] XMBWDFGMSWQBCA-RNFDNDRNSA-M 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 230000008961 swelling Effects 0.000 claims description 6
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 6
- JBWKIWSBJXDJDT-UHFFFAOYSA-N triphenylmethyl chloride Chemical compound C=1C=CC=CC=1C(C=1C=CC=CC=1)(Cl)C1=CC=CC=C1 JBWKIWSBJXDJDT-UHFFFAOYSA-N 0.000 claims description 6
- 238000010521 absorption reaction Methods 0.000 claims description 5
- 239000007795 chemical reaction product Substances 0.000 claims description 4
- 239000008055 phosphate buffer solution Substances 0.000 claims description 3
- QRUDEWIWKLJBPS-UHFFFAOYSA-N benzotriazole Chemical compound C1=CC=C2N[N][N]C2=C1 QRUDEWIWKLJBPS-UHFFFAOYSA-N 0.000 claims description 2
- 238000001514 detection method Methods 0.000 claims description 2
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 claims description 2
- 239000008363 phosphate buffer Substances 0.000 claims description 2
- 239000007790 solid phase Substances 0.000 claims description 2
- 210000004027 cell Anatomy 0.000 abstract description 45
- 238000003384 imaging method Methods 0.000 abstract description 22
- 102000004190 Enzymes Human genes 0.000 abstract description 19
- 108090000790 Enzymes Proteins 0.000 abstract description 19
- 239000002086 nanomaterial Substances 0.000 abstract description 16
- 239000002243 precursor Substances 0.000 abstract description 16
- 230000002285 radioactive effect Effects 0.000 abstract description 10
- 230000000694 effects Effects 0.000 abstract description 9
- 206010028980 Neoplasm Diseases 0.000 abstract description 7
- 238000000338 in vitro Methods 0.000 abstract description 7
- 238000001338 self-assembly Methods 0.000 abstract description 7
- 238000003745 diagnosis Methods 0.000 abstract description 3
- 210000004881 tumor cell Anatomy 0.000 abstract description 3
- 201000010099 disease Diseases 0.000 abstract description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 2
- 102000004196 processed proteins & peptides Human genes 0.000 abstract 2
- 230000008685 targeting Effects 0.000 abstract 1
- 238000002474 experimental method Methods 0.000 description 24
- 229910052739 hydrogen Inorganic materials 0.000 description 22
- 239000001257 hydrogen Substances 0.000 description 18
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 14
- 230000004700 cellular uptake Effects 0.000 description 11
- 239000002105 nanoparticle Substances 0.000 description 11
- 238000001228 spectrum Methods 0.000 description 11
- 238000005406 washing Methods 0.000 description 10
- 150000003384 small molecules Chemical class 0.000 description 8
- 241001465754 Metazoa Species 0.000 description 7
- 238000006482 condensation reaction Methods 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 230000003834 intracellular effect Effects 0.000 description 5
- 230000003211 malignant effect Effects 0.000 description 5
- 238000009833 condensation Methods 0.000 description 4
- 238000002296 dynamic light scattering Methods 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 3
- 230000005540 biological transmission Effects 0.000 description 3
- 229910002091 carbon monoxide Inorganic materials 0.000 description 3
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000012404 In vitro experiment Methods 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 230000005494 condensation Effects 0.000 description 2
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 2
- 239000010931 gold Substances 0.000 description 2
- 229910052737 gold Inorganic materials 0.000 description 2
- 230000012447 hatching Effects 0.000 description 2
- 150000002431 hydrogen Chemical class 0.000 description 2
- 238000001819 mass spectrum Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000002096 quantum dot Substances 0.000 description 2
- 238000010200 validation analysis Methods 0.000 description 2
- 238000009736 wetting Methods 0.000 description 2
- DEWLEGDTCGBNGU-UHFFFAOYSA-N 1,3-dichloropropan-2-ol Chemical compound ClCC(O)CCl DEWLEGDTCGBNGU-UHFFFAOYSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- YYQRGCZGSFRBAM-UHFFFAOYSA-N Triclofos Chemical compound OP(O)(=O)OCC(Cl)(Cl)Cl YYQRGCZGSFRBAM-UHFFFAOYSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 238000000701 chemical imaging Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 238000013016 damping Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000013399 early diagnosis Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 125000003827 glycol group Chemical group 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 229940089468 hydroxyethylpiperazine ethane sulfonic acid Drugs 0.000 description 1
- 239000012216 imaging agent Substances 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000005311 nuclear magnetism Effects 0.000 description 1
- 238000011580 nude mouse model Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000002797 proteolythic effect Effects 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 229960001147 triclofos Drugs 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention discloses a single photon emission computed tomography developer and a preparation method thereof. The method is characterized by respectively synthetising two peptides by the solid-phase synthesis to obtain corresponding initial products and final products, synthetising a single compound with a first kind feature structure (I) or a second kind feature structure (II) and forming the developer according to a needed dosage ratio. The imaging developer is micro-molecule, good in hydrophilicity and easy to prepare, and precursor micro-molecule of the developer has peptides specially recognized by specific enzyme, so that imaging areas can be targeted actively, and controlled self-assembly of radioactive nano-structures in living cells is realized to study activity of specific enzymes in tumor cells. Compared with conventional micro-nano systems used in early disease diagnosis, by the preparation method, micro-nano materials synthetised in vitro are not needed to be injected into living bodies for tumor diagnosis, problems of low intake and difficulty in targeting of conventional nano-materials are solved.
Description
Technical field
The invention belongs to nanometer developer technical field, be specifically related to nanometer developer of single photon emission computerized tomography and preparation method thereof.
Background technology
Britain < < nature > > magazine biotechnology periodical (Nat.Biotechnol., 2004, Vol.22 (8), P.969) a kind of quantum dot nano preparation for bio-imaging and preparation method thereof is disclosed, although this preparation has good imaging effect, and can resist photobleaching, but because first this preparation of preparation wants synthesized semiconductor nano-quantum point, then not only to modify polyoxyethylene glycol fragment and improve the metabolism cycle, also to modify the antibody of target specific recognition, so it is more difficult that preparation is got up, and can cause toxicity because of the contained heavy metal of this class quantum dot itself, also limited the development of this class nanometer preparation.The periodical < < of GDCh applied chemistry > > (Angew.Chem.Int.Ed., 2008, Vol.47 (15), P.2804) introduced a kind of " switching mode " near infrared gold nano preparation and preparation method thereof, although this preparation has good biocompatibility, but because it is that optical molecular is modified to gold nano surface, also to modify the peptide section of target specific recognition, synthetic more loaded down with trivial details, stability neither be fine.In a word, the shortcoming such as it is high that existing these nanometer developer ubiquities are prepared difficulty, and cellular uptake rate is low, target is difficult, forms the bottleneck into restriction biology imaging research.
Summary of the invention
The object of the invention is to propose a kind of single photon emission computerized tomography developer and preparation method thereof, to obtain the radioactive nano developer of the real-time controlled self-assembly of energy, overcoming traditional nanometer developer, to prepare difficulty high, the shortcomings such as cellular uptake rate is low, target is difficult, can be used for the activity of certain enzyme in study tumor cell, thereby can implement non-intruding, carry out intuitively, fast the early diagnosis of disease.
The preparation method of single photon emission computerized tomography developer of the present invention, is characterized in that:
The first step, first adopt solid phase synthesis process synthetic two sections of peptide chains respectively, process is as follows: by 1 mmole 2-chlorine trityl chloride resin at 4-6 milliliter N, in dinethylformamide after swelling 4-8 minute, add first amino acid tyrosine of 2 mmoles, add again 2 mmole N, N-diisopropylethylamine, react after 2-3 hour, with 100 microliter methanol reaction 10-20 minute, cut the protecting group of tyrosine, second the amino acid cysteine reaction 2-3 hour of 1.6 mmole that adds activation, cut the protecting group of halfcystine, the 3rd the amino acids Arginine reaction 2-3 hour of 1.6 mmoles that adds activation, cut arginic protecting group, the 4th the amino acids Arginine reaction 2-3 hour of 1.6 mmoles that adds activation, cut arginic protecting group, the 1.6 mmole five amino acid α-amino-isovaleric acid reaction 2-3 hour that add activation, cut the protecting group of α-amino-isovaleric acid, the 6th the amino acids Arginine reaction 3-4 hour of 1.6 mmoles that adds again activation, cut the protecting group of last amino acids Arginine, add 2-3 mmole aceticanhydride reaction 20-30 minute, finally with the methylene dichloride of the trifluoroacetic acid that is 1% containing volumetric concentration, from this resin, cut above-mentioned synthetic peptide section, add again ether, on refrigerated centrifuge, upper strata ether is removed in centrifugal layering hypsokinesis, treat that the dry resulting white solid powder afterwards of solvent evaporates is tyrosine-halfcystine-arginine-arginine-α-amino-isovaleric acid-arginine peptide section, again by 1 mmole 2-chlorine trityl chloride resin at 4-6 milliliter N, in dinethylformamide after swelling 4-8 minute, add first amino acid α-amino-isovaleric acid of 2 mmoles, add again 2 mmole N, N-diisopropylethylamine, react after 2-3 hour, with 100 microliter methanol reaction 10-20 minute, cut the protecting group of α-amino-isovaleric acid, second the amino acids Arginine reaction 2-3 hour of 1.6 mmole that adds activation, cut arginic protecting group, the 3rd the amino acid cysteine reaction 2-3 hour of 1.6 mmoles that adds activation, cut the protecting group of halfcystine, the 4th the amino acids Arginine reaction 2-3 hour of 1.6 mmoles that adds activation, cut arginic protecting group, the 1.6 mmole five amino acid tyrosine reaction 2-3 hour that add activation, cut the protecting group of tyrosine, the 6th the amino acids Arginine reaction 3-4 hour of 1.6 mmoles that adds again activation, cut the protecting group of last amino acids Arginine, add 2-3 mmole aceticanhydride reaction 20-30 minute, the dichloromethane solution of the trifluoroacetic acid that is finally 1% by volumetric concentration cuts above-mentioned synthetic peptide section from this resin, add again ether, on refrigerated centrifuge, upper strata ether is removed in centrifugal layering hypsokinesis, treat that the dry resulting white solid powder afterwards of solvent evaporates is α-amino-isovaleric acid-arginine-halfcystine-arginine-tyrosine-arginine peptide section,
Second step, above-mentioned synthetic tyrosine-halfcystine-arginine-arginine-α-amino-isovaleric acid-arginine peptide section is got to 0.1 mmole be dissolved in 2 milliliters of tetrahydrofuran solvents, add 0.15 mmole N-methylmorphine, while being cooled to zero degree, add 0.12 mmole isobutyl chlorocarbonate, activate after half an hour, add 0.12 mmole 2-amino-6-cyano group benzothiazole, maintain the temperature at 0-10 ℃ one hour, stirring at room reaction is spent the night, through high performance liquid chromatography separating-purifying, being collected in ultraviolet 320 nanometers has the component of strong absorption, is first kind head product; Again above-mentioned synthetic α-amino-isovaleric acid-arginine-halfcystine-arginine-tyrosine-arginine peptide section being got to 0.1 mmole is dissolved in 2 milliliters of tetrahydrofuran solvents, add 0.15 mmole N-methylmorphine, while being cooled to zero degree, add 0.12 mmole isobutyl chlorocarbonate, activate after half an hour, add 0.12 mmole 2-amino-6-cyano group benzothiazole, maintain the temperature at 0-10 ℃ one hour, stirring at room reaction is spent the night, through high performance liquid chromatography separating-purifying, being collected in ultraviolet 320 nanometers has the component of strong absorption, is Equations of The Second Kind head product;
The 3rd step, first the first kind head product of above-mentioned preparation is dissolved in to stirring reaction 3-5 hour in the dichloromethane solution of the trifluoroacetic acid that 10 ml volumes concentration are 95%, through high performance liquid chromatography separating-purifying, being collected in ultraviolet 320 nanometers has strong absorbent components to be end product structural formula to be
the first compounds X
1or structural formula is
the third compounds X
3; Equations of The Second Kind head product is dissolved in to stirring reaction 3-5 hour in the dichloromethane solution of the trifluoroacetic acid that 10 ml volumes concentration are 95%, through high performance liquid chromatography separating-purifying, being collected in ultraviolet 320 nanometers has strong absorbent components to be end product structural formula to be again
the 5th kind of compound Y
1or structural formula is
the 7th kind of compound Y
3;
The 4th step, employing chloramine-t method mark radioiodine, process is as follows: by 50 microlitre concentration, be first 5mg/mL
Structural formula is
the first compounds X
1add in reaction flask injection 1mCi
125i-NaI, the chloramine-T that adds 15 microlitre 2mg/mL reacts one minute, then adds the sodium metabisulphite termination reaction of 20 microlitre 2mg/mL, obtains corresponding structural formula be with the high performance liquid chromatography separation that is equipped with γ-detector
the second compounds X
2; By 50 microlitre concentration, be that 5mg/mL structural formula is again
the first compounds X
1add reaction flask
In, injection 1mCi
131i-NaI, the chloramine-T that adds 15 microlitre 2mg/mL reacts one minute, then adds the sodium metabisulphite termination reaction of 20 microlitre 2mg/mL, obtains corresponding structural formula be with the high performance liquid chromatography separation that is equipped with γ-detector
the 4th kind of compounds X
4; And then the structural formula that is 5mg/mL by 50 microlitre concentration is
the 5th kind of compound Y
1add in reaction flask injection 1mCi
125i-NaI, the chloramine-T that adds 15 microlitre 2mg/mL reacts one minute, then adds the sodium metabisulphite termination reaction of 20 microlitre 2mg/mL, and wherein institute's reagent adding is to be all dissolved in the phosphate buffer solution of pH=7.4; With the high performance liquid chromatography separation that is equipped with γ-detector, obtaining corresponding structural formula is
the 6th kind of compound Y
2; By 50 microlitre concentration, be finally that 5mg/mL structural formula is
the 5th kind of compound Y
1add in reaction flask injection 1mCi
131i-NaI, the chloramine-T that adds 15 microlitre 2mg/mL reacts one minute, then adds the sodium metabisulphite termination reaction of 20 microlitre 2mg/mL, obtains corresponding structural formula be with the high performance liquid chromatography separation that is equipped with γ-detector
the 8th kind of compound Y
4; The structural formula wherein relating in the 4th step is
the first compounds X
1, structural formula is
the 5th kind of compound Y
1, chloramine-T, sodium metabisulphite be to be all dissolved in 0.05mol/L according to each self-corresponding mass body volume concentrations above, in the phosphate buffer soln of pH=7.4, join in reaction system;
Wherein solid phase amino acid used is all with 9-fluorenylmethyloxycarbonyl
as alpha-amino group protecting group, the sulfydryl of halfcystine is protected by tertiary butylthio, arginine side chain amino by
protection; When the selected tyrosine of synthetic peptide chain is
time, what the 3rd step obtained is exactly that structural formula is
the third compounds X
3with structural formula be
the 7th kind of compound Y
3, when the selected tyrosine of synthetic peptide chain is
time, what the 3rd step obtained is exactly that structural formula is
the first compounds X
1with structural formula be
the 5th kind of compound Y
1; Wherein the reagent of activated amino acid is I-hydroxybenzotriazole and the benzotriazole-N with the amount concentration of amino acid commaterial, N, N', N'-tetramethyl-urea hexafluorophosphate; When often connecting an amino acid and cutting protecting group, all to test with Caesar (kaiser test) reagent detection imino-and whether exist: if positive aobvious blue, show to have sheared; If negative displaing yellow, shows that amino acid connects.
Thus, in the preparation method of above-mentioned single photon emission computerized tomography developer of the present invention, synthesized following eight kinds of concrete single compounds with first kind feature structure (I) or Equations of The Second Kind feature structure (II):
Radicals R in structural formula above
1be selected from H or Ac, R
2be selected from H or
125i or I or
131i, R
3be selected from H or Ac, R
4be selected from H or
125i or I or
131i.
The single compound wherein with first kind feature structure (I) is:
The first compounds X
1: the radicals R in its structural formula
1and R
2be all H;
The second compounds X
2: the radicals R in its structural formula
1for H and R
2for
125i;
The third compounds X
3: the radicals R in its structural formula
1for Ac and R
2for I;
The 4th kind of compounds X
4: the radicals R in its structural formula
1for H and R
2for
131i;
The single compound with Equations of The Second Kind feature structure (II) is:
The 5th kind of compound Y
1: the radicals R in its structural formula
3and R
4be all H;
The 6th kind of compound Y
2: the radicals R in its structural formula
3for H and R
4for
125i;
The 7th kind of compound Y
3: the radicals R in its structural formula
3for Ac and R
4for I;
The 8th kind of compound Y
4: the radicals R in its structural formula
3for H and R
4for
131i.
The described compound with first kind feature structure (I) is to be sheared the compound that self-assembly condensation reaction occurs by enzyme spcificity identification, described in there is Equations of The Second Kind feature structure (II) thus compound be to be identified the control compound that self-assembling reaction can not occur by enzyme; The second compounds X
2, the 4th kind of compounds X
4, the 6th kind of compound Y
2with the 8th kind of compound Y
4contain radioiodine; The third compounds X
3with the 7th kind of compound Y
3contain cold natural iodine, it is in order to improve the chemical concentrations of preparation with corresponding radioactive compound co-cultivation in experiment; The first compounds X
1, the second compounds X
2, the third compounds X
3for doing single photon emission computerized tomography precursor cell picked-up experiment; The third compounds X
3with the 4th kind of compounds X
4for doing single photon emission computerized tomography experimentation on animals; The 5th kind of compound Y
1, the 6th kind of compound Y
2, the 7th kind of compound Y
3for doing single photon emission computerized tomography precursor cell picked-up control experiment; The 7th kind of compound Y
3with the 8th kind of compound Y
4for doing the control experiment of single photon emission computerized tomography animal.
Single photon emission computerized tomography developer of the present invention, it is characterized in that comprising by suitable dosage ratio by the third compounds X
3with the 4th kind of compounds X
4the video picture group mixture forming, and supporting by the 7th kind of compound Y with it
3with the 8th kind of compound Y
4the control group mixture forming; Described is to guarantee the optimum quantity that tells on when using this preparation in different experiments object and clinical experiment by suitable dosage ratio, is specially: the third compounds X
3with the 7th kind of compound Y
3concentration must be respectively between 100 μ mol/L to 200 μ mol/L, the 4th kind of compounds X
4with the 8th kind of compound Y
4radioactive dosage should be respectively between 2.5mCi/L to 4mCi/L.
The product that adopts the inventive method to prepare, wherein, except for developer, also comprises by the third compounds X
3with the second compounds X
2the precursor research mixture mix forming and with it supporting the 7th kind of compound Y
3with the 6th kind of compound Y
2the single photon emission computerized tomography precursor developer that mixes the control group mixture forming, is characterized in that comprising the third compounds X by suitable dosage ratio
3with the second compounds X
2the mixture forming, and the 7th kind of compound Y
3with the 6th kind of compound Y
2the control group mixture forming; Described is to guarantee the optimum quantity that tells on during for different experimental conditions by suitable dosage ratio, is specially: the third compounds X
3with the 7th kind of compound Y
3concentration must reach respectively between 100 μ mol/L to 200 μ mol/L, the second compounds X
2with the 6th kind of compound Y
2radioactive dosage should be respectively between 1mCi/L to 5mCi/L.
Compare with traditional small molecules preparation, the significant advantage of nanometer imaging developer of the present invention is the nano material that is self-assembled in target spot place intelligence by small molecules, at imaging region, accumulate, and reach good gathering, expand effect, thereby it is not high to have overcome small molecules sensitivity, plasma half-life is short, uses the shortcoming that needs high density, has greatly improved imaging sensitivity; Different from traditional nanometer preparation is, because the precursor of imaging developer of the present invention is small molecules, wetting ability is fine, preparation easily, then can fix a point to arrive video picture region and be self-assembled into nano particle, therefore imaging developer of the present invention has overcome again a difficult problem for general nano material preparation difficulty, low picked-up and target difficulty when possessing the advantage of nano material.Because the precursor small molecules of nanometer imaging developer of the present invention has the peptide section of certain enzyme specific recognition, therefore targeted imaging region initiatively, is different from general nano particle by the enhancing infiltration of tumor tissues and keeps effect to reach the object of passive target imaging region.Therefore, utilize single photon emission computerized tomography developer of the present invention and preparation method thereof can controlled self-assembly radioactive nano structure, for the imaging research of certain enzyme activity.In a word, adopt the inventive method not need to synthesize in vitro micro-nano material and reinject in vivo for the diagnosis of tumour, overcome the low picked-up of traditional micro Nano material and a difficult problem for target difficulty.This is to utilize the inventive method live body to prepare in real time the intelligence of Nano medication.Can also be for the proteolytic ferment design precursor compound of some malignant tumour high expression level, thereby at the real-time self-assembled nano structures of tumor locus, nano material can be used as a carrier and loads a large amount of chemical imaging agent effectively to improve the preparation concentration of biological tissue part, and this is the target intelligence of utilizing the inventive method.The research work of this respect at present there is not yet bibliographical information.To sum up, the present invention has obvious novelty and creativeness.
Accompanying drawing explanation
Fig. 1 is the first compounds X synthetic in embodiment 1
1nucleus magnetic hydrogen spectrum figure;
Fig. 2 is the third compounds X synthetic in embodiment 1
3nucleus magnetic hydrogen spectrum figure;
Fig. 3 is the 5th kind of compound Y synthetic in embodiment 1
1nucleus magnetic hydrogen spectrum figure;
Fig. 4 is the 7th kind of compound Y in embodiment synthetic 1
3nucleus magnetic hydrogen spectrum figure.
Fig. 5 is the third compounds X in embodiment 2
3the transmission electron microscope that Condensation in Vitro forms nanoparticle characterizes;
Fig. 6 is the third compounds X in embodiment 2
3condensation in Vitro forms the dynamic light scattering analysis of nanoparticle;
Fig. 7 is the third compounds X in embodiment 2
3vitro enzyme is cut the uv-absorbing figure at 500-700nm of front and back.
Fig. 8 is the second compounds X in embodiment 3
2with the 6th kind of compound Y
2the comparison of cellular uptake;
Fig. 9 is the second compounds X in embodiment 3
2respectively at the third compounds X of different concns
3(0,25,50,100 μ mol/L) hatches 30min jointly, when said mixture is 160min from the time washing out in cell and the comparison of residual quantity in cell afterwards;
Figure 10 is the second compounds X in embodiment 3
2with the 6th kind of compound Y
2having and nothing 100 the third compounds Xs of μ mol/L respectively
3time and the 7th kind of compound Y
3jointly hatch after 30min with cell, from the time plot washing out in cell;
Figure 11 is the second compounds X in embodiment 3
2with the 6th kind of compound Y
2respectively having, without 100 the third compounds Xs of μ mol/L
3time and the 7th kind of compound Y
3jointly hatch 30min, when said mixture is 160min from the time washing out in cell and the comparison of residual quantity in cell afterwards.
Embodiment
The synthetic of probe is provided in the following examples 1, and embodiment 2 is Validation in vitro experiment, and embodiment 3 is cellular uptake experiment.The described compound with first kind feature structure (I) is to be sheared the compound that self-assembly condensation reaction occurs by enzyme spcificity identification, described in there is Equations of The Second Kind feature structure (II) thus compound be to be identified the control compound that self-assembling reaction can not occur by enzyme.The second compounds X
2, the 4th kind of compounds X
4, the 6th kind of compound Y
2with the 8th kind of compound Y
4contain radioiodine, the third compounds X
3with the 7th kind of compound Y
3contain cold natural iodine, be in order to improve the chemical concentrations of preparation with corresponding radioactive compound co-cultivation in experiment.The first compounds X
1, the second compounds X
2, the third compounds X
3for doing single photon emission computerized tomography precursor cell picked-up experiment; The third compounds X
3with the 4th kind of compounds X
4for doing single photon emission computerized tomography experimentation on animals; The 5th kind of compound Y
1, the 6th kind of compound Y
2, the 7th kind of compound Y
3for doing single photon emission computerized tomography precursor cell picked-up control experiment; The 7th kind of compound Y
3with the 8th kind of compound Y
4for doing the control experiment of single photon emission computerized tomography animal.
Embodiment 1:
First adopt solid-phase synthesis that tyrosine-halfcystine-arginine-arginine-α-amino-isovaleric acid-arginine and two peptide sections of α-amino-isovaleric acid-arginine-halfcystine-arginine-tyrosine-arginine are synthesized respectively.
The second compounds X in the present embodiment
2the synthetic route of (Acetyl-Arg-Val-Arg-Arg-Cys (StBu)-Tyr (I-125)-CBT) is as follows:
First adopt solid-phase synthesis that tyrosine-halfcystine-arginine-arginine-α-amino-isovaleric acid-arginine peptide section is synthesized, concrete steps are as follows: by 0.33 mmole 2-chlorine trityl chloride resin at 2 milliliters of N, in dinethylformamide, swelling, after five minutes, adds first amino acid of 0.66 mmole in reactor
add 0.66 mmole N, N-diisopropylethylamine, react after two hours, with 30 microliter methanol reaction 20 minutes, cut first amino acid protecting group, kaiser test is aobvious blue, add second amino acid cysteine of 0.55 mmole of activation to react two hours, kaiser tests displaing yellow, cut protecting group, kaiser test is aobvious blue, add the 3rd amino acids Arginine of 0.55 mmole of activation to react three hours, kaiser tests displaing yellow, cut protecting group, kaiser test is aobvious blue, add the 4th amino acids Arginine of 0.55 mmole of activation to react three hours, kaiser tests displaing yellow, cut protecting group, kaiser test is aobvious blue, add 0.55 mmole five amino acid α-amino-isovaleric acid of activation to react two hours, kaiser tests displaing yellow, cut protecting group, kaiser test is aobvious blue, add the 6th amino acids Arginine of 0.55 mmole of activation to react four hours, kaiser tests displaing yellow, cut protecting group, kaiser test is aobvious blue, add 1 mmole aceticanhydride reaction 20 minutes, finally with the methylene dichloride that containing volumetric concentration is 1% trifluoroacetic acid, from resin, cut peptide section, add again ether, on refrigerated centrifuge, upper strata ether is removed in centrifugal layering hypsokinesis, after solvent evaporates is dry, obtain white solid powder and be will be synthetic peptide section, above-mentioned synthetic peptide section is taken to 0.1 mmole to be dissolved in 2 milliliters of tetrahydrofuran solvents, add 0.15 mmole N-methylmorphine, while being cooled to zero degree, add 0.12 mmole isobutyl chlorocarbonate, after half an hour, add 0.12 mmole 2-amino-6-cyano group benzothiazole, maintain the temperature at 0 ℃ to 10 ℃ one hour, stirring at room reaction is spent the night, and through high performance liquid chromatography separating-purifying, being collected in ultraviolet 320 nanometers has strong absorbent components to obtain head product B, then the head product B obtaining in previous step is dissolved in the methylene dichloride of the trifluoroacetic acid that 10 ml volumes concentration are 95% to stirring at room reaction 3 hours, through high performance liquid chromatography separating-purifying, being collected in ultraviolet 320 nanometers has strong absorbent components to be the first compounds X
1.After above-mentioned reactions steps, also will be with chloramine-t method mark radioiodine, process is as follows: first by 50 microlitres, 5mg/mL the first compounds X
1add in reaction flask injection 1mCi
125i-NaI, adds 15 microlitres, 2mg/mL chloramine-T reaction one minute, add 20 microlitres, 2mg/mL sodium metabisulphite termination reaction, wherein above-mentioned institute reagent adding is to be all dissolved in the phosphate buffer solution of pH=7.4, with the high performance liquid chromatography separation that is equipped with γ-detector, obtains the second compounds X
2.
Adopt German Brooker company (bruker) Brooker nuclear-magnetism software to resolve four kinds of compounds synthetic in the present embodiment and obtain four kinds of nucleus magnetic hydrogen spectrum figure as shown in Figures 1 to 4:
Fig. 1 is the first compounds X synthetic in the present embodiment
1nucleus magnetic hydrogen spectrum figure; Fig. 2 is the third compounds X
3nucleus magnetic hydrogen spectrum figure; Fig. 3 is the 5th kind of compound Y
1nucleus magnetic hydrogen spectrum figure; Fig. 4 is the 7th kind of compound Y
3nucleus magnetic hydrogen spectrum figure.The first compounds X
1molecular formula is C
49h
74n
18o
8s
3, [(M+H)
+]: 1139.51, gained mass spectrum is m/z1139.3.From accompanying drawing 1, the first compounds X
1nucleus magnetic hydrogen spectrum (d-methyl alcohol, 300MHz): 8.63 (s, 1H), 8.13 (d, J=9.0Hz, 1H), 7.70 (d, J=9.0Hz, 1H), 7.10 (d, J=9.0Hz, 2H) these are fragrant ring hydrogen, 6.69 (d, J=9.0Hz, 2H), 4.68 (t, J=7.6Hz, 1H), 4.58 (q, J=5Hz, 1H), 4.34 (m, 3H), 4.16 (d, J=7.3Hz, 1H), 3.14-3.25 (m, 8H), 3.10 (d, J=5.4Hz, 1H), 2.96-3.06 (m, 2H), 2.04 (s, 3H) be the hydrogen of Ac, 1.58-1.95 (br, 14H), 1.32 (s, 20H), 0.98 (d, J=6.7Hz, 6H) be two methyl hydrogen of α-amino-isovaleric acid.
The 7th kind of compound Y
3the synthetic route of (Acetyl-Arg-Tyr (Ac) (I)-Arg-Cys (StBu)-Arg-Val-CBT) is as follows:
Adopt again solid-phase synthesis that α-amino-isovaleric acid-arginine-halfcystine-arginine-tyrosine-arginine peptide section is synthesized, concrete steps are as follows: by 0.33 mmole 2-chlorine trityl chloride resin at 2 milliliters of N, in dinethylformamide, swelling is after five minutes, in reactor, add first amino acid α-amino-isovaleric acid of 0.66 mmole, add 0.66 mmole N, N-diisopropylethylamine, react after two hours, with 30 microliter methanol reaction 20 minutes, cut first amino acid protecting group, kaiser test is aobvious blue, add second amino acids Arginine of 0.55 mmole of activation to react three hours, kaiser tests displaing yellow, cut protecting group, kaiser test is aobvious blue, add the 3rd amino acid cysteine of 0.55 mmole of activation to react three hours, kaiser tests displaing yellow, cut protecting group, kaiser test is aobvious blue, add the 4th amino acids Arginine of 0.55 mmole of activation to react three hours, kaiser tests displaing yellow, cut protecting group, kaiser test is aobvious blue, add 0.55 mmole five amino acid tyrosine of activation to react three hours, kaiser tests displaing yellow, cut protecting group, kaiser test is aobvious blue, add the 6th amino acids Arginine of 0.55 mmole of activation to react four hours, kaiser tests displaing yellow, cut protecting group, kaiser test is aobvious blue, add 1 mmole aceticanhydride reaction 20 minutes, finally with the methylene dichloride that containing volumetric concentration is 1% trifluoroacetic acid, from resin, cut peptide section, add again ether, on refrigerated centrifuge, upper strata ether is removed in centrifugal layering hypsokinesis, after solvent evaporates is dry, obtaining white solid powder is α-amino-isovaleric acid-arginine-halfcystine-arginine-tyrosine-arginine peptide section, above-mentioned synthetic peptide section is taken to 0.1 mmole to be dissolved in 2 milliliters of tetrahydrofuran solvents, add 0.15 mmole N-methylmorphine, while being cooled to zero degree, add 0.12 mmole isobutyl chlorocarbonate, after half an hour, add 0.12 mmole 2-amino-6-cyano group benzothiazole, maintain the temperature at 0 ℃ to 10 ℃ one hour, stirring at room reaction is spent the night, and through high performance liquid chromatography separating-purifying, being collected in ultraviolet 320 nanometers has strong absorbent components to obtain head product K, then the head product K obtaining in previous step is dissolved in the methylene dichloride of the trifluoroacetic acid that 10 ml volumes concentration are 95% to stirring reaction 3 hours, through high performance liquid chromatography separating-purifying, being collected in ultraviolet 320 nanometers has strong absorbent components to be the 7th kind of compound Y
3.
The 7th kind of compound Y
3molecular formula is C
51h
76iN
18o
9s
3, [(M+H)+]: 1307.42, gained mass spectrum is m/z1307.2.From 4, the seven kinds of compound Y of accompanying drawing
3nucleus magnetic hydrogen spectrum (d-methyl alcohol, 300MHz): 8.65 (d, J=2Hz, 1H), 8.13 (d, J=9Hz, 1H), 7.75 (dd, J
1=9Hz, J
2=2Hz, 2H), 7.28 (dd, J
1=8.5Hz, J
2=2.2Hz, 1H), 7.02 (d, J=8.2Hz, 1H) these are fragrant ring hydrogen, 4.54-4.68 (br, 3H), 4.40-4.48 (m, 1H), 4.37 (d, J=7.1Hz, 1H), 4.23-4.32 (m, 1H), 4.11-4.19 (m, 1H), 3.53-3.60 (br, 1H), 2.98-3.26 (br, 15H), 2.33 (s, 3H) are the hydrogen of Ac on tyrosine phenyl ring, 2.03 (s, 3H) are the hydrogen of the arginine end-blocking Ac of place, 1.48-1.77 (br, 13H), 1.24-1.43 (m, 33H), two methyl hydrogen that 1.05 (d, J=6.8Hz, 7H) are α-amino-isovaleric acid.
Thus, in the above-mentioned preparation process of the present embodiment, synthesized following eight kinds of concrete single compounds with first kind feature structure (I) or Equations of The Second Kind feature structure (II):
Radicals R in structural formula above
1be selected from H or Ac, R
2be selected from H or
125i or
131i, R
3be selected from H or Ac, R
4be selected from H or
125i or
131i.
The single compound wherein with first kind feature structure (I) is:
The first compounds X
1: the radicals R in its structural formula
1and R
2be all H;
The second compounds X
2: the radicals R in its structural formula
1for H and R
2for
125i;
The third compounds X
3: the radicals R in its structural formula
1for Ac and R
2for I;
The 4th kind of compounds X
4: the radicals R in its structural formula
1for H and R
2for
131i;
The single compound with Equations of The Second Kind feature structure (II) is:
The 5th kind of compound Y
1: the radicals R in its structural formula
3and R
4be all H;
The 6th kind of compound Y
2: the radicals R in its structural formula
3for H and R
4for
125i;
The 7th kind of compound Y
3: the radicals R in its structural formula
3for Ac and R
4for I;
The 8th kind of compound Y
4: the radicals R in its structural formula
3for H and R
4for
131i.
The described compound with first kind feature structure (I) is to be sheared the compound that self-assembly condensation reaction occurs by enzyme spcificity identification, described in there is Equations of The Second Kind feature structure (II) thus compound be to be identified the control compound that self-assembling reaction can not occur by enzyme; The second compounds X
2, the 4th kind of compounds X
4, the 6th kind of compound Y
2with the 8th kind of compound Y
4contain radioiodine; The third compounds X
3with the 7th kind of compound Y
3contain cold natural iodine, it is in order to improve the chemical concentrations of preparation with corresponding radioactive compound co-cultivation in experiment; The first compounds X
1, the second compounds X
2, the third compounds X
3for doing single photon emission computerized tomography precursor cell picked-up experiment; The third compounds X
3with the 4th kind of compounds X
4for doing single photon emission computerized tomography experimentation on animals; The 5th kind of compound Y
1, the 6th kind of compound Y
2, the 7th kind of compound Y
3for doing single photon emission computerized tomography precursor cell picked-up control experiment; The 7th kind of compound Y
3with the 8th kind of compound Y
4for doing the control experiment of single photon emission computerized tomography animal.
Single photon emission computerized tomography developer of the present invention, comprise by suitable dosage ratio by the third compounds X
3with the 4th kind of compounds X
4the video picture group mixture forming, and supporting by the 7th kind of compound Y with it
3with the 8th kind of compound Y
4the control group mixture forming; Described is to guarantee the optimum quantity that tells on when using this preparation in different experiments object and clinical experiment by suitable dosage ratio, is specially: the third compounds X
3with the 7th kind of compound Y
3concentration must be respectively between 100 μ mol/L to 200 μ mol/L, the 4th kind of compounds X
4with the 8th kind of compound Y
4radioactive dosage should be respectively between 2.5mCi/L to 4mCi/L.
The product that adopts aforesaid method to prepare, wherein except for developer, also comprise by suitable dosage ratio by the third compounds X
3with the second compounds X
2the precursor research mixture mix forming and with it supporting the 7th kind of compound Y
3with the 6th kind of compound Y
2the single photon emission computerized tomography precursor developer that mixes the control group mixture forming; Described is to guarantee the optimum quantity that tells on during for different experimental conditions by suitable dosage ratio, is specially: the third compounds X
3with the 7th kind of compound Y
3concentration must reach respectively between 100 μ mol/L to 200 μ mol/L, the second compounds X
2with the 6th kind of compound Y
2radioactive dosage should be respectively between 1mCi/L to 5mCi/L.
Embodiment 2: Validation in vitro experiment
What in the present embodiment experiment in vitro, adopt is the third compounds X
3concentration is 100 μ mol/L, the cumulative volume that is the not woods enzyme of 100mmol/L4-hydroxyethyl piperazine ethanesulfonic acid damping fluid, 1mmol/L trichloroethyl phosphate, 1mmol/L calcium chloride and 5 μ L in concentration is in the solution of 50 μ L, hatch for 30 ℃ and be condensed into dimer in 16 hours, and be self-assembled into nanoparticle.
Fig. 5 has provided the third compounds X in the present embodiment
3the transmission electron microscope that Condensation in Vitro forms nanoparticle characterizes; Fig. 6 is the third compounds X in the present embodiment
3condensation in Vitro forms the dynamic light scattering analysis of nanoparticle; Fig. 7 is the third compounds X in embodiment 2
3vitro enzyme is cut front and back at the uv-absorbing figure of 500-700nm.
Wherein Fig. 5 is transmission electron microscope (TEM) sign of the nanoparticle that preparation forms to the inventive method, and the mean sizes of its nanoparticle of product that employing the inventive method prepares is 415nm.Fig. 6 is dynamic light scattering (DLS) analysis, shows that its mean sizes is 363-429nm.Fig. 7 is the variation of enzyme-added front and back uv-absorbing within the scope of 500-700nm, in Fig. 7, can find out, top enzyme is cut previous curve b and is shown that it does not absorb at 500-700nm, below enzyme is cut curve a afterwards and is shown that it has absorption at 500-700nm, and this has shown the generation of condensation reaction and the formation of nanoparticle.
Embodiment 3: cellular uptake experiment
First cultivate malignant galactophore JEG-3 MDA-MB-468, concrete operations are as follows: malignant galactophore JEG-3 MDA-MB-468 is used the DMEM culture medium culturing containing the bovine serum albumin of volumetric concentration 10%, and cell is being 5%CO containing volumetric concentration
237 ℃ of thermostat containers of air ambient in cultivate, every one day, change a subculture.
Do cellular uptake experiment, concrete operations are as follows again: by malignant galactophore JEG-3 MDA-MB-468 kind in six orifice plates, about 1,000,000 cells in each hole, volume is 1mL, described is 5%CO containing volumetric concentration in the above
237 ℃ of constant incubators of air ambient in cultivate six hours after, the second compounds Xs that contain 1 μ Ci are changed in three holes wherein
2fresh culture, and another three holes are changed to the 6th kind of compound Y that contains 1 μ Ci
2fresh culture, then every 5,10,20,30 and 60min, collect part substratum, by the substratum of gamma counter measurement collection and the radioactive intensity of cell, calculate uptake ratio.
Finally do cell and wash out experiment, concrete operations are as follows: by malignant galactophore JEG-3 MDA-MB-468 kind in six orifice plates, about 1,000,000 cells in each hole, described is in the above 5%CO containing volumetric concentration
237 ℃ of constant incubators of air ambient in cultivate six hours after, will in first three hole of first block of plate, be replaced by the second compounds X that contains 1 μ Ci of 1mL
2fresh culture, will in its another three holes, be replaced by the 6th kind of compound Y that contains 1 μ Ci of 1mL
2fresh culture; The second compounds X that contains 1 μ Ci of 1mL will be replaced by first three hole of second block of plate
2the third compounds X with 25 μ mol/L
3fresh culture, will in other three holes of second block of plate, be replaced by the second compounds X that contains 1 μ Ci of 1mL
2the third compounds X with 50 μ mol/L
3fresh culture; The second compounds X that contains 1 μ Ci of 1mL will be replaced by first three hole of the 3rd block of plate
2the third compounds X with 100 μ mol/L
3fresh culture, will in other three holes of the 3rd block of plate, be replaced by the 6th kind of compound Y that contains 1 μ Ci of 1mL
2the 7th kind of compound Y with 100 μ mol/L
3fresh culture.Cultivating after half an hour, is 5,10,20,40,80 and during 160min in the time, more renews substratum and collects former substratum, and when 160min, want collecting cell simultaneously, finally uses the radioactive intensity of gamma counter measurement collection substratum and cell.
Fig. 8 is the second compounds X in embodiment 3
2with the 6th kind of compound Y
2the comparison of cellular uptake; Fig. 9 is the second compounds X in embodiment 3
2respectively at the third compounds X of different concns
3(0,25,50,100 μ mol/L) hatches 30min jointly, when said mixture is 160min from the time washing out in cell and the comparison of residual quantity in cell afterwards; Figure 10 is the second compounds X in embodiment 3
2with the 6th kind of compound Y
2having and nothing 100 the third compounds Xs of μ mol/L respectively
3time and the 7th kind of compound Y
3jointly hatch after 30min with cell, from the time plot washing out in cell; Figure 11 is the second compounds X in embodiment 3
2with the 6th kind of compound Y
2respectively having, without 100 the third compounds Xs of μ mol/L
3time and the 7th kind of compound Y
3jointly hatch 30min, when said mixture is 160min from the time washing out in cell and the comparison of residual quantity in cell afterwards.
As shown in Figure 8, square point curve in the top is the second compounds X
2cellular uptake curve c, middle circle point curve is the 6th kind of compound Y
2cellular uptake curve d, bottom trigonometric curve is two kinds of compound picked-up difference curve e.By to the second compounds X
2with the 6th kind of compound Y
2the comparison of cellular uptake, shows the second compounds X
2have than the 6th kind of compound Y
2high cellular uptake, and when hatching 30min left and right, cellular uptake reaches capacity.Then further condition is groped.Fig. 9 is the second compounds X in embodiment 3
2respectively at the third compounds X of different concns
3(0,25,50,100 μ mol/L) hatches after 30min jointly, when said mixture is 160min from the time washing out in cell and the comparison of residual quantity in cell afterwards.By 1 μ Ci the second compounds X
2the third compounds X with different concns
3jointly hatch after 30min with cell, intracellular radioactivity residual quantity when relatively this mixture is 160min from the time washing out in cell and afterwards, left narrow column with slant lines f, left wide column with slant lines g are respectively the third compounds Xs
3cell radioactivity residual quantity while being 0 μ mol/L and 25 μ mol/L, showing that cell radioactivity is residual does not have a considerable change; Right narrow column with slant lines h is the third compounds X
3cell radioactivity residual quantity when concentration is 50 μ mol/L, shows the residual obvious raising of radioactivity; Right wide column with slant lines i is the third compounds X
3cell radioactivity residual quantity when concentration is 100 μ mol/L, its radioactivity is residual is as seen from Figure 9 the second compounds X
2while hatching separately (left narrow column with slant lines f) nearly four times, therefore, acquired results proves the third compounds X
3when concentration need to reach 100 μ mol/L, could there is condensation reaction and effectively improve radioactivity the second compounds X
2in intracellular picked-up.
Figure 10 is the second compounds X in embodiment 3
2with the 6th kind of compound Y
2having and nothing 100 the third compounds Xs of μ mol/L respectively
3time and the 7th kind of compound Y
3jointly hatch after 30min with cell, from the time plot washing out in cell.As shown in Figure 10, by 1 μ Ci the second compounds X
2with the 6th kind of compound Y of 1 μ Ci
2respectively with the third compounds X of different concns
3(the corresponding curve m of 0 μ mol/L, the corresponding curve k of 100 μ mol/L) and the 7th kind of compound Y
3(curve l during 0 μ mol/L, curve j during 100 μ mol/L) hatches after 30min jointly with cell, the Washout Curve in 160min, and wherein in figure, minimum three curve l, j, m is respectively the 6th kind of compound Y of 1 μ Ci
2, the 6th kind of compound Y of 1 μ Ci
2the 7th kind of compound Y with 100 μ mol/L
3mixture, 1 μ Ci the second compounds X
2jointly hatch after 30min the Washout Curve in 160min with cell; The highest curve k is 1 μ Ci the second compounds X
2the third compounds X with 100 μ mol/L
3jointly hatch after 30min the Washout Curve in 160min with cell.Relatively cell wash-in goes out intracellular radioactivity residual quantity afterwards, can obviously find out 1 μ Ci the second compounds X
2with 100 the third compounds Xs of μ mol/L
3have higher residual.
Figure 11 is the second compounds X in embodiment 3
2with the 6th kind of compound Y
2respectively having, without 100 the third compounds Xs of μ mol/L
3time and the 7th kind of compound Y
3jointly hatch 30min, when said mixture is 160min from the time washing out in cell and the comparison of residual quantity in cell afterwards.As shown in Figure 11, left black post n, left column with slant lines o represent respectively 1 μ Ci the second compounds X
2, 1 μ Ci the second compounds X
2with 100 the third compounds Xs of μ mol/L
3jointly hatch and wash out after 160min residual quantity in cell; Right black post p, right column with slant lines q represent respectively the 6th kind of compound Y of 1 μ Ci
2, the 6th kind of compound Y of 1 μ Ci
2with the 7th kind of compound Y of 100 μ mol/L
3jointly hatch and wash out after 160min residual quantity in cell.As can be seen from Fig. 11, the height of left column with slant lines o is high nearly five times of other three posts, shows 1 μ Ci the second compounds X
2with 100 the third compounds Xs of μ mol/L
3(left column with slant lines o) jointly hatched and washed out after 160min residual quantity in cell is 1 μ Ci the second compounds X
2hatch separately nearly five times, 1 the 6th kind of μ Ci compound Y
2with the 7th kind of compound Y
3at 0 micromole (right black post p) and the right column with slant lines q of 100 μ mol/L() jointly hatch comparison acquired results and be not improved, and lower than 1 μ Ci the second compounds X
2hatch separately the intracellular residual quantity of (left black post n).Therefore prove the third compounds X
3concentration need to reach 100 μ mol/L and condensation reaction just can occur and effectively improved radioactive compound the second compounds X
2in intracellular picked-up.
After carrying out cellular uptake experiment, do exactly experimentation on animals.First by malignant galactophore cancer cells MDA-MB-468(high expression level woods enzyme not) on the nude mice of heavily about 20g, set up tumor model.Treat that tumor growth is to 300-700mm
3size, tail vein is given 2 the third compounds Xs of μ mol
3with the 4th kind of compounds X of 50 μ Ci
4.Compound arrives after tumor locus, is absorbed and in cell, is self-assembled into nanostructure " intelligence ", and do SPECT video picture by tumour cell.Simultaneously by the 7th kind of compound Y of 2 μ mol
3with the 8th kind of compound Y of 50 μ Ci
4mixture carries out contrast experiment.According to known this preparation of cell experiment result, there is good effect.
Known by above-described embodiment and result thereof: nanometer imaging developer of the present invention is the nano material that is self-assembled in target spot place intelligence by small molecules, at imaging region, accumulate, thereby can reach good gathering, expand effect, greatly improved imaging sensitivity; Because the precursor of imaging developer of the present invention is small molecules, wetting ability is fine, preparation easily, then can fix a point to arrive video picture region and be self-assembled into nano particle, therefore imaging developer of the present invention has overcome again a difficult problem for general nano material preparation difficulty, low picked-up and target difficulty when possessing the advantage of nano material.Because the precursor small molecules of nanometer imaging developer of the present invention has the peptide section of certain enzyme specific recognition, so can active targeted imaging region.To sum up, utilize single photon emission computerized tomography developer of the present invention and preparation method thereof can controlled self-assembly radioactive nano structure, for the imaging research of certain enzyme activity.
Claims (3)
1. a preparation method for single photon emission computerized tomography developer, is characterized in that:
The first step, first adopt solid phase synthesis process synthetic two sections of peptide chains respectively, process is as follows: by 1 mmole 2-chlorine trityl chloride resin at 4-6 milliliter N, in dinethylformamide after swelling 4-8 minute, add first amino acid tyrosine of 2 mmoles, add again 2 mmole N, N-diisopropylethylamine, react after 2-3 hour, with 100 microliter methanol reaction 10-20 minute, cut the protecting group of tyrosine, second the amino acid cysteine reaction 2-3 hour of 1.6 mmole that adds activation, cut the protecting group of halfcystine, the 3rd the amino acids Arginine reaction 2-3 hour of 1.6 mmoles that adds activation, cut arginic protecting group, the 4th the amino acids Arginine reaction 2-3 hour of 1.6 mmoles that adds activation, cut arginic protecting group, the 1.6 mmole five amino acid α-amino-isovaleric acid reaction 2-3 hour that add activation, cut the protecting group of α-amino-isovaleric acid, the 6th the amino acids Arginine reaction 3-4 hour of 1.6 mmoles that adds again activation, cut the protecting group of last amino acids Arginine, add 2-3 mmole aceticanhydride reaction 20-30 minute, finally with the methylene dichloride of the trifluoroacetic acid that is 1% containing volumetric concentration, from this resin, cut above-mentioned synthetic peptide section, add again ether, on refrigerated centrifuge, upper strata ether is removed in centrifugal layering hypsokinesis, treat that the dry resulting white solid powder afterwards of solvent evaporates is tyrosine-halfcystine-arginine-arginine-α-amino-isovaleric acid-arginine peptide section, again by 1 mmole 2-chlorine trityl chloride resin at 4-6 milliliter N, in dinethylformamide after swelling 4-8 minute, add first amino acid α-amino-isovaleric acid of 2 mmoles, add again 2 mmole N, N-diisopropylethylamine, react after 2-3 hour, with 100 microliter methanol reaction 10-20 minute, cut the protecting group of α-amino-isovaleric acid, second the amino acids Arginine reaction 2-3 hour of 1.6 mmole that adds activation, cut arginic protecting group, the 3rd the amino acid cysteine reaction 2-3 hour of 1.6 mmoles that adds activation, cut the protecting group of halfcystine, the 4th the amino acids Arginine reaction 2-3 hour of 1.6 mmoles that adds activation, cut arginic protecting group, the 1.6 mmole five amino acid tyrosine reaction 2-3 hour that add activation, cut the protecting group of tyrosine, the 6th the amino acids Arginine reaction 3-4 hour of 1.6 mmoles that adds again activation, cut the protecting group of last amino acids Arginine, add 2-3 mmole aceticanhydride reaction 20-30 minute, the dichloromethane solution of the trifluoroacetic acid that is finally 1% by volumetric concentration cuts above-mentioned synthetic peptide section from this resin, add again ether, on refrigerated centrifuge, upper strata ether is removed in centrifugal layering hypsokinesis, treat that the dry resulting white solid powder afterwards of solvent evaporates is α-amino-isovaleric acid-arginine-halfcystine-arginine-tyrosine-arginine peptide section,
Second step, above-mentioned synthetic tyrosine-halfcystine-arginine-arginine-α-amino-isovaleric acid-arginine peptide section is got to 0.1 mmole be dissolved in 2 milliliters of tetrahydrofuran solvents, add 0.15 mmole N-methylmorphine, while being cooled to zero degree, add 0.12 mmole isobutyl chlorocarbonate, activate after half an hour, add 0.12 mmole 2-amino-6-cyano group benzothiazole, maintain the temperature at 0-10 ℃ one hour, stirring at room reaction is spent the night, through high performance liquid chromatography separating-purifying, being collected in ultraviolet 320 nanometers has the component of strong absorption, is first kind head product; Again above-mentioned synthetic α-amino-isovaleric acid-arginine-halfcystine-arginine-tyrosine-arginine peptide section being got to 0.1 mmole is dissolved in 2 milliliters of tetrahydrofuran solvents, add 0.15 mmole N-methylmorphine, while being cooled to zero degree, add 0.12 mmole isobutyl chlorocarbonate, activate after half an hour, add 0.12 mmole 2-amino-6-cyano group benzothiazole, maintain the temperature at 0-10 ℃ one hour, stirring at room reaction is spent the night, through high performance liquid chromatography separating-purifying, being collected in ultraviolet 320 nanometers has the component of strong absorption, is Equations of The Second Kind head product;
The 3rd step, first the first kind head product of above-mentioned preparation is dissolved in to stirring reaction 3-5 hour in the dichloromethane solution of the trifluoroacetic acid that 10 ml volumes concentration are 95%, through high performance liquid chromatography separating-purifying, being collected in ultraviolet 320 nanometers has strong absorbent components to be end product structural formula to be
the first compounds X
1or structural formula is
the third compounds X
3; Equations of The Second Kind head product is dissolved in to stirring reaction 3-5 hour in the dichloromethane solution of the trifluoroacetic acid that 10 ml volumes concentration are 95%, through high performance liquid chromatography separating-purifying, being collected in ultraviolet 320 nanometers has strong absorbent components to be end product structural formula to be again
the 5th kind of compound Y
1or structural formula is
the 7th kind of compound Y
3;
The 4th step, employing chloramine-t method mark radioiodine, process is as follows: by 50 microlitre concentration, be first that 5mg/mL structural formula is
the first compounds X
1add in reaction flask injection 1mCi
125i-NaI, the chloramine-T that adds 15 microlitre 2mg/mL reacts one minute, then adds the sodium metabisulphite termination reaction of 20 microlitre 2mg/mL, obtains corresponding structural formula be with the high performance liquid chromatography separation that is equipped with γ-detector
the second compounds X
2; By 50 microlitre concentration, be that 5mg/mL structural formula is again
the first compounds X
1add in reaction flask injection 1mCi
131i-NaI, the chloramine-T that adds 15 microlitre 2mg/mL reacts one minute, then adds the sodium metabisulphite termination reaction of 20 microlitre 2mg/mL, obtains corresponding structural formula be with the high performance liquid chromatography separation that is equipped with γ-detector
the 4th kind of compounds X
4; And then the structural formula that is 5mg/mL by 50 microlitre concentration is
the 5th kind of compound Y
1add in reaction flask injection 1mCi
125i-NaI, the chloramine-T that adds 15 microlitre 2mg/mL reacts one minute, then adds the sodium metabisulphite termination reaction of 20 microlitre 2mg/mL, and wherein institute's reagent adding is to be all dissolved in the phosphate buffer solution of pH=7.4; With the high performance liquid chromatography separation that is equipped with γ-detector, obtaining corresponding structural formula is
the 6th kind of compound Y
2; By 50 microlitre concentration, be finally that 5mg/mL structural formula is
the 5th kind of compound Y
1add in reaction flask injection 1mCi
131i-NaI, the chloramine-T that adds 15 microlitre 2mg/mL reacts one minute, then adds the sodium metabisulphite termination reaction of 20 microlitre 2mg/mL, obtains corresponding structural formula be with the high performance liquid chromatography separation that is equipped with γ-detector
the 8th kind of compound Y
4; The structural formula wherein relating in the 4th step is
the first compounds X
1, structural formula is
the 5th kind of compound Y
1, chloramine-T, sodium metabisulphite be to be all dissolved in 0.05mol/L according to each self-corresponding mass body volume concentrations above, in the phosphate buffer soln of pH=7.4, join in reaction system; Wherein solid phase amino acid used is all with 9-fluorenylmethyloxycarbonyl
as alpha-amino group protecting group, the sulfydryl of halfcystine is protected by tertiary butylthio, arginine side chain amino by
protection; When the selected tyrosine of synthetic peptide chain is
time, what the 3rd step obtained is exactly that structural formula is
the third compounds X
3with structural formula be
the 7th kind of compound Y
3, when the selected tyrosine of synthetic peptide chain is
time, what the 3rd step obtained is exactly that structural formula is
the first compounds X
1with structural formula be
the 5th kind of compound Y
1; Wherein the reagent of activated amino acid is I-hydroxybenzotriazole and the benzotriazole-N with the amount concentration of amino acid commaterial, N, N', N'-tetramethyl-urea hexafluorophosphate; When often connecting an amino acid and cutting protecting group, all to test with Caesar (kaiser test) reagent detection imino-and whether exist: if positive aobvious blue, show to have sheared; If negative displaing yellow, shows that amino acid connects.
2. the single photon emission computerized tomography developer that described in claim 1 prepared by method, is characterised in that it is: structural formula is
the 4th kind of compounds X
4.
3. a single photon emission computerized tomography developer mixture, is characterized in that by having structural formula being
the third compounds X
3with structural formula be
the 4th kind of compounds X
4mix and form, described the third compounds X
3concentration between 100 μ mol/L to 200 μ mol/L, described the 4th kind of compounds X
4radioactive dosage between 2.5mCi/L to 4mCi/L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210164547.6A CN102690328B (en) | 2012-05-24 | 2012-05-24 | Single photon emission computed tomography developer and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210164547.6A CN102690328B (en) | 2012-05-24 | 2012-05-24 | Single photon emission computed tomography developer and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102690328A CN102690328A (en) | 2012-09-26 |
| CN102690328B true CN102690328B (en) | 2014-09-24 |
Family
ID=46856080
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210164547.6A Expired - Fee Related CN102690328B (en) | 2012-05-24 | 2012-05-24 | Single photon emission computed tomography developer and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102690328B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105085617B (en) * | 2014-08-20 | 2019-08-13 | 江苏省原子医学研究所 | A kind of compound and the preparation method and application thereof for nanometer PET imaging agent |
| CN104177475B (en) * | 2014-08-21 | 2016-09-14 | 南京医科大学 | A kind of19f NMR probe and preparation method thereof |
| CN104650181B (en) * | 2015-03-10 | 2017-07-21 | 上海久裕生物科技有限公司 | One class supramolecular hydrogel gel nanometer materials and gelator precursor and its preparation method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1493566A (en) * | 2003-09-15 | 2004-05-05 | 江苏省原子医学研究所 | Preparation method of 1,4,7,10,13-penta azacyclopentadecane |
| CN1528744A (en) * | 2003-10-17 | 2004-09-15 | 江苏省原子医学研究所 | Positron emitted computerized tomography imaging agent benzamide derivative synthesizing method |
| CN101157644A (en) * | 2007-11-16 | 2008-04-09 | 江苏省原子医学研究所 | Method for preparing adamantine disulfide tetroxide derivative |
-
2012
- 2012-05-24 CN CN201210164547.6A patent/CN102690328B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1493566A (en) * | 2003-09-15 | 2004-05-05 | 江苏省原子医学研究所 | Preparation method of 1,4,7,10,13-penta azacyclopentadecane |
| CN1528744A (en) * | 2003-10-17 | 2004-09-15 | 江苏省原子医学研究所 | Positron emitted computerized tomography imaging agent benzamide derivative synthesizing method |
| CN101157644A (en) * | 2007-11-16 | 2008-04-09 | 江苏省原子医学研究所 | Method for preparing adamantine disulfide tetroxide derivative |
Non-Patent Citations (2)
| Title |
|---|
| 张荣军等.肝ASGP受体功能显像剂的制备及显像研究.《核技术》.2000,第23卷(第11期),第779页. |
| 肝ASGP受体功能显像剂的制备及显像研究;张荣军等;《核技术》;20001231;第23卷(第11期);第779页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102690328A (en) | 2012-09-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Bhatt et al. | Recent advances in zirconium-89 chelator development | |
| Zhu et al. | Aggregation-induced emission: a trailblazing journey to the field of biomedicine | |
| CN102827252B (en) | One kind of magnetic resonance imaging developers and two-photon imaging developers and preparation method thereof | |
| Yuan et al. | GSH-activated light-up near-infrared fluorescent probe with high affinity to αvβ3 integrin for precise early tumor identification | |
| CN104749377B (en) | A kind of fluorescent probe with aggregation-induced emission characteristic and its preparation method and application | |
| CN106967152B (en) | A kind of compound and the preparation method and application thereof of Value linear label | |
| CN106470964A (en) | There is the application in the treatment of imaging and imaging guiding of the polymer of aggregation-induced emission property and oligomer | |
| CN102690328B (en) | Single photon emission computed tomography developer and preparation method thereof | |
| CN103030682B (en) | Enzyme-sensitive supramolecular hydrogel nano material, gelator and preparation method of gelator | |
| CN104174039A (en) | Nanosilicon dioxide/ferroferric oxide shell-core particle surface modifying method and magnetic nano material | |
| CN105419781A (en) | Click chemistry-based tumor marker probe, preparation method and application | |
| CN109096170A (en) | Nir dye, its targeted imaging agent, nano-carrier and anticancer drug and application | |
| Usama et al. | Targeted dual-modal PET/SPECT-NIR imaging: from building blocks and construction strategies to applications | |
| CN107778367B (en) | Polypeptide, application thereof and medicine containing polypeptide | |
| CN104749369A (en) | Fluorescent labeling method for living organisms having cell membrane structures | |
| CN103784978B (en) | A kind of albumen-dye composition and application thereof | |
| Guillou et al. | Photoactivatable fluorescent tags for dual-modality positron emission tomography optical imaging | |
| CN108314678B (en) | Using phosphatidylserine as molecular probe of target spot and application thereof | |
| CN103143037B (en) | Method for synthesizing rare earth metal compound nano cluster and application thereof | |
| CN105884867B (en) | 18The affinity body class compound and the preparation method and application thereof of F label | |
| CN104177475B (en) | A kind of19f NMR probe and preparation method thereof | |
| Wang et al. | " Click Chemistry" for Molecular Imaging | |
| Krishnan et al. | Agarose Micro‐Well Platform for Rapid Generation of Homogenous 3D Tumor Spheroids | |
| CN105085617B (en) | A kind of compound and the preparation method and application thereof for nanometer PET imaging agent | |
| Krishnan et al. | Developing μSpherePlatform using a commercial hairbrush: an agarose 3d culture platform for deep-tissue imaging of prostate cancer |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140924 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |