CN107496943A - The preparation method for the Dimer San A Cyclopeptide derivatives cancer of pancreas molecular probes that F 18 is marked - Google Patents
The preparation method for the Dimer San A Cyclopeptide derivatives cancer of pancreas molecular probes that F 18 is marked Download PDFInfo
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Abstract
The present invention relates to a kind of preparation method of the Dimer San A Cyclopeptide derivatives cancer of pancreas molecular probes of F 18 marks, this method comprises the following steps：(1) prepare18F normal saline solutions：By Na18F solution is enriched in Sep Park light QMA posts；Then elute and collect through deionized water, physiological saline, obtain18F normal saline solutions；(2) acetic acid, AlCl3And18Reaction solution is obtained after the heating of F normal saline solutions；(3) acetonitrile is added in NOTA Dimer Sansalvamide A polypeptides, fully dissolved after ultrasonic oscillation, add deionized water, the polypeptide dissolved；The polypeptide of dissolving is moved in reaction solution, heated, cooling, obtains label18F‑NOTA‑Dimer‑Sansalvamide A；(4) label18F NOTA Dimer San dissolve after removing solvent through phosphate buffer, membrane filtration, produce18The Dimer Sansalvamide A Cyclopeptide derivatives cancer of pancreas molecular probes of F marks.The present invention first using NOTA Dimer San molecules as18The precursor of F marks, the target spot using Hsp90 as imaging, structure18The new developers of NOTA Dimer San of F marks, the developer have the advantage for making a distinction cancer of pancreas and pancreatitis.
Description
Technical field
The present invention relates to a kind of preparation method of probe, more particularly to the Dimer-San A Cyclopeptide derivatives of F-18 marks
The preparation method of cancer of pancreas molecular probe.
Background technology
Conventional structure image（Including CT, MRI and ultrasonic examination）Without specificity；Molecular image（molecular
imaging）It is to show tissue level, cell, the specific molecular of subcellsular level with iconography means, reacts under condition of living organism
The change of molecular level, so as to carry out the research of qualitative and quantitative to disease so that " the precisely diagnosis " of disease is possibly realized.
PET/CT (positron e mission computed tomography) is the important imaging device of molecular image, its appearance, is medical science shadow
As the revolution again learned, generally acknowledged and extensive concern by medical field, survey titles " hat of modern high technology ", be described as detecting and swell
" radar " of knurl, " Holmes " of diagnosing tumor.PET and CT are perfectly combined in one by PET/CT, by PET metabolic characteristics
It is combined with the ripe accurate anatomical informations of CT, the two mutual supplement with each other's advantages, 1+1 can be reached in medical diagnosis on disease>2 effect.But with
Conventional image is a difference in that PET imagings need to rely on internal injection developer.
The presence of positive electron or " positive electron " are proved in the 1930s.When positive electron and electron interaction,
It is equal, in opposite direction that they launch two energy in annihilation process（1800）Photon.PET is one kind positron radioactivity
The radioactive tracer imaging technique that nucleic is carried out, after entering in vivo using the various positron emitting tracers of injection, determine live body group
Knit change, acceptor distribution, the combination of internal antigen-antibody, weary oxygen, blood perfusion and gene expression of intracellular various metabolism etc.
Situation, and intuitively shown with image format, once check and be achieved with whole body fault image, PET images the work that is otherwise known as
The biochemical imaging of body, molecular imaging.PET imagings are available for tumour, central nervous system, three big field of heart, wherein with tumour
Clinical and basic research is in the highest flight.
The available positron emitting tracers of PET have a lot.18F- deoxyglucoses (18F-FDG it is) most widely used at present
General positron emitting tracer.18F-FDG PET be respectively to the average sensitivity of diagnosis of malignant tumor and specificity 84% and
88%, changed the therapeutic scheme of 30% patient.Thus,18F-FDG PET are some in assessment18The high intakes of F-FDG
It has been a ripe method in terms of malignant tumour.
18F is a kind of nucleic for launching positive electron, and half-life period is 110 min, is appropriate for PET, PET/CT imaging.20
The twenties in century, German biochemist find that glycolysis is one main metabolic pathway of tumour cell by zoopery, swollen
Oncocyte is to provide the energy needed for its fast breeding, utilization of the tumour cell to glucose by ATP caused by glycolysis
Greatly increase.18F-FDG PET and18F-FDG PET/CT imagings are namely based on this principle, and FDG structure is similar to grape
Sugar, one of oh group are substituted by a F atom.Cell starts the sugar similar to glucose to FDG capture process
Glycolysis process, entered through GLUT with speed K1 mediation cross-films in cell liquid, under the catalytic action of hexokinase,
6- phosphoric acid-FDG (FDG-6-PO are turned into speed K3 FDG phosphorylations4), but with the phosphoric acid Portugal of metabolin 6 of natural glucose
Grape sugar is different, FDG-6-PO4It is no longer participate in further glycometabolism process, the phosphatase in cytosol can be by 6- phosphoric acid-FDG
Removing phosphoric acid turns into FDG, then from being mediated positioned at the GLUT of cell membrane inner surface with speed K2 to extracellular transport.It is but most of to utilize
Phosphatase content is very low in the more tissue of glucose, and 6- phosphoric acid-FDG can not pass through cell membrane diffusion so that and 6- phosphoric acid-
FDG goes the reverse transformation of phosphoric acid very low with the speed for leaving cell, is trapped in cell and is imaged as tracer, FDG
Dense poly- amount and the metaboilic level of glucose in the cell is proportionate.
18F- deoxyglucoses (18F-FDG) it is the most frequently used global PET radiopharmaceutical at present, is by Ido etc. 1978
Synthesize first.18F has the ability combined with glucose molecule, and it instead of hydrogen atom or hydroxyl, without significantly changing glucose
The biological function of molecule.In addition, its half-life period is relatively long（110 min）.Glycolysis is to produce ATP basic bioid
Approach, it is generally present in normal cell under the conditions of weary oxygen.However, in most of cancer cells, even in oxygen-enriched ring
There is also glycolysis in border, and tumor cells showed glucose consumption is elevated.Therefore,18F- deoxyglucoses (18F-
FDG it is) developer the most frequently used global PET/CT at present, is a kind of tumour non-specificity developer.This spy for coming from medicine
The design method of pin is a kind of method of high risk and high reward.Because additional label is likely to affect the life of medicine in itself
Thing activity, the failure for causing probe to design, if but the medicine remains to keep its bioactivity to keep constant, the spy after mark
The possibility that pin is transformed into clinical practice in the future is very big.
18Prepared by F-FDG passes through automatic chemistry synthesis module（Chemistry process control unit,
CPCU）, it is automatically synthesized using software control.Synthetic method is with tetra--oxy-acetyls of 1,3,4,6- -2- oxygen-fluoroform sulphonyl
Base-β-D- mannopyranoses（Abbreviation mannose triflate）For raw material, in phase transfer catalyst Kryptofix2.2.2 (amino-polyethers
2.2.2 under) promoting, 18With the hydroxyl on mannose triflate 2 nucleophilic substitution, generation occur for F ion18F-FDG protection types
Precursor, through acid or basic hydrolysis slough acetyl protection base and obtain18F-FDG。18F-FDG PET/CT are non-specific as a kind of tumour
Property developer, obvious deficiency in terms of tumour early stage malignant and benign lesion diagnosis be present：False positive and false negative are simultaneously deposited, so as to cause
The mistaken diagnosis of benign from malignant tumors diagnosis is with failing to pinpoint a disease in diagnosis.
Cancer of pancreas is the fifth-largest malignant tumour in the world, and its median survival interval is less than 6 months, and 5 years total survival rates are less than 6%,
Only 10% patient is adapted to operative treatment, and its poor prognosis and the disease fail to early diagnose closely related.
SUVmax (Standard Uptake Value, standardization intake maximum) is weigh glycometabolism degree half
Quantitative target.The metabolism hyperplasia Showed Very Brisk of most of malignant cells, GLUT in malignant cell
MRNA up-regulated expressions:The horizontal rise of GLUT Glut1 and Glut3；The intracellular horizontal rise of hexokinase；Grape
Sugar -6- phosphatase levels are lowered:So that anaerobic glycolysis metabolic rate substantially increases in malignant cell, show in aerobic ring
Anerobic glycolysis feature under border, make FDG in tumour cell it is dense it is poly- be higher than normal cell, so as to distinguish it is benign and malignant carefully
Difference of Metabolism in born of the same parents, find to be metabolized vigorous malignant tumor tissue.But for cancer of pancreas, glycometabolism, which increases, not to be had
Specificity；On the contrary, even if glycometabolism is not high, the diagnosis of cancer of pancreas can not be excluded completely.
The antidiastole of cancer of pancreas and mass pancreatitis is that current clinical medicine and medical imaging are urgently to be resolved hurrily
Common difficulty, the two shows hypermetabolism in PET image, and the extent of metabolism of the latter even can exceed the former, therefore the two is in PET
Findings exist overlapping, certain difficulty is brought to antidiastole.
Meta analysis and research displays：18Sensitivity, the specificity of F-FDG PET/CT antidiastoles cancers of pancreas and pancreatitis
Respectively 90%, 84%.There is scholar to marked the molecular image diagnosis that CA199 antibody fragments carry out cancer of pancreas, but radio-immunity shows
As whether use radioactive nuclear mark monoclone antibody or its fragment, there is that molecular weight is larger, and blood is removed slowly
（4~20h）, the problem of being difficult to obtain higher T/NT ratios within a short period of time；Furthermore CA199 is cancer of pancreas non-specificity
Antigen, also there is obvious expression in pancreas inflammatory lesion.Exploring the PET developers of new diagnosing tumour Just because of this just turns into
Instantly the focus studied.
Tagged ligand molecular weight used in rii receptor is small, blood removes that fast, tissue penetration is strong, T/NT ratios are high, nothing
Immunogenicity, have the advantages that high sensitivity, high specificity and accuracy are good, be the most active forward position research neck of molecular nuclear medicine
One of domain.
Heat shock protein 90（Heart shock protein 90, Hsp90）It is a kind of highly conserved general in living nature
Store-through, have the albumen of particular molecule chaperone function, molecular weight is about 83 ~ 90KDa.Hsp90 contains three highly conserved knots
Structure domain：That is N-terminal atriphos binding domain, central domain, C-terminal domain, in the form of homodimer exist, mainly with
Cell cycle is related to apoptosis regulation.Research confirms that Hsp90 is in the Several Kinds of Malignancy cell including cancer of pancreas
Overexpression, it is 2 ~ 10 times of normal cell, and generation with tumour, development, classification, by stages and prognosis is closely related；
Hsp90 is activated in tumour cell matter and navigates to cell surface, and is only resided in normal cell in cytoplasm.Cause
This, Hsp90 is of increasing concern as a potential oncotherapy research target spot, also turns into the molecular image of target for it and studies
Lay a good foundation.
Ogata, M. etc. compare Pancreatic Adenocarcinoma, control group（Pancreatitis and normal pancreatic tissue）Hsp90 expression
Situation, as a result show：Hsp90 α mRNA prompt cancer of pancreas group in the overexpression of Pancreatic Adenocarcinoma apparently higher than control group, the result
It is related to cell propagation to knit Hsp90 α mRNA.But Hsp90 β overexpression is in Pancreatic Adenocarcinoma, pancreatitis and Normal Pancreas
Tissue almost uniformly expression.
Sansalvamide A (abbreviation San A) be 1999 by U.S. Belofsky etc. from ocean Pseudomonas Fusarium
In separate a kind of be made up of two leucines, a valine, a phenylalanine and an Alpha-hydroxy isocaproic acid
Ring pentapeptide ester type compound, there is very high lipophilicity and significant anti-tumor capacity, it is to national cancer institute
60 kinds of cancer cell lines have a significant antiproliferative activity, and to including pancreatic carcinoma in interior kinds of tumor cells
With treatment targeting.Research is found, substitutes phenyl ring in San A cyclic peptide molecules to align hydrogen using groups such as fluorine, chlorine, methoxyl groups
Atom, the anti-tumor biological of gained San A Cyclopeptide derivatives are substantially better than San A, have unique anticancer property, and
There is no structural homology with the medicine of current anti-pancreatic cancer.Up to the present, the San A Cyclopeptide derivatives of synthesis have more than 100
Kind, its antitumor activity difference is obvious.
The content of the invention
The technical problems to be solved by the invention are to provide a kind of Dimer-San for the F-18 marks for effectively improving mark rate
The preparation method of A Cyclopeptide derivatives cancer of pancreas molecular probes.
To solve the above problems, the Dimer-San A Cyclopeptide derivatives cancer of pancreas molecules of F-18 marks of the present invention
The preparation method of probe, comprises the following steps：
(1) prepare18F- normal saline solutions：
50 ~ 100mCi Na that cyclotron is produced18F solution be first enriched in respectively through the mol/L of 10 mL 0.5 and pH=
8.4 NaOAc solution and the Sep-Park light QMA posts of 10 mL deionized waters pretreatment；Then with 5 mL deionized waters
Elution removes metal impurities ion；Eluted and collected with the mL physiological saline of 0.2 mL ~ 1 again, obtained18F- normal saline solutions；
(2) μ L of acetic acid 5,0.01 M AlCl are added in 5mL vials312 μ L's and 10 ~ 20 mCi is described18F- gives birth to
The μ L of saline solution 100 are managed, 10 min are heated at a temperature of 120 DEG C, obtain reaction solution；
(3) the μ g of NOTA-Dimer-Sansalvamide A polypeptides 100 ~ 150 are weighed in 1.5 mL EP pipes, add 250 ~ 350
μ L acetonitriles, fully dissolve after the min of ultrasonic oscillation 2, add 40 μ L deionized waters, the polypeptide dissolved；The dissolving
Polypeptide move in the reaction solution, cool down 5 min after 10 min are heated at a temperature of 100 ~ 105 DEG C, obtain label18F-
NOTA-Dimer-Sansalvamide A, referred to as18F-NOTA-Dimer-San, its structural formula are as follows：
；
(4) the label18F-NOTA-Dimer-San removes solvents with dry machine is hanged in 40 DEG C, and with the phosphorus containing 5% DMSO
Phthalate buffer dissolves, and is most produced afterwards through 0.22 μm of membrane filtration18The Dimer-Sansalvamide A cyclic peptide of F marks derives
Thing cancer of pancreas molecular probe.
The condition of the step (3) middle supersonic oscillations refers to that temperature is 37 DEG C, and the concussion time is 2 min.
The step (4) middle phosphate buffer and the label18F-NOTA-Dimer-San ratio is 300 ~ 500
μL：100~150 μg.
The present invention has advantages below compared with prior art：
1st, the precursor that is marked first using NOTA-Dimer-San molecules as positron-emitting radionuclides of the present invention, using Hsp90 as
The target spot of imaging, structure18The new developers of NOTA-Dimer-San of F marks.
The present invention is with the most strong cyclic peptide Dimer-Sansalvamide A (letters of activity in Sansalvamide A derivatives
Claim Dimer-San A) it is used as labelled precursor (IC50For 1-20 nM), its structural formula is as follows：
The labelled precursor is on the premise of the analog derivative important activity functional group is retained, by Dimer-San A molecules
Phenyl ring on introduce amino, couple Isosorbide-5-Nitrae, the azo-cycle nonanes of 7- tri--Isosorbide-5-Nitrae, 7- triacetic acids（Isosorbide-5-Nitrae, 7-triazacyclononane-1,
4,7-triacetic acid, NOTA）Bifunctional chelating agent, after carrying out identification of its biological activity to it, realize positron emission
Property nucleic18F carries out indirect labelling to it（18F-NOTA-Dimer-San, referred to as18F-NOTA-Dimer-San）.Research card
In fact, San A and its Cyclopeptide derivatives are Hsp90 inhibitor, N-terminal and middle destructing domain of its selective binding in Hsp90, are led to
Cross and prevent Hsp90 C-terminal domain from being combined with client protein downstream, be related to cell growth and tumor signal biography so as to disturb
The multiple paths led, cause apoptosis of tumor cells, and the two is combined with high specific, high-affinity.
2nd, developer of the present invention18Target spots of the F-NOTA-San using Hsp90 as imaging, experiment card is absorbed through cell in vitro
Real, it has higher Percentage bound with target spot, and cells blocks test the specificity that further demonstrate the developer and targeted integration
（Referring to Fig. 1 ~ 9）；Build nude mice cancer of pancreas and carry out Micro-PET imagings with aseptic inflammation model, it is seen that tumor developer is taken the photograph
Take substantially, and inflammatory model corresponding site has no intake, therefore developer cancer of pancreas height absorbs, and inflammatory lesions do not absorb, and borrow
This can make a distinction cancer of pancreas and lump pancreatitis, can overcome conventional developer18F-FDG deficiency, shows itself
Advantage.
【Cell in vitro absorbs and blocking experiment】
PL45 cells are collected and planted to 24 orifice plates（0.5×105Cells/well）, put to the h of incubate box culture 24 (37oC-,
5% CO2)。
Non-blacked experiment adds the developer of same concentrations per hole（5 μCi/100 μL）；Kong Xianjia for blocking experiment
Enter the certain density non-marked peptides of 100 μ L, the developer of same concentrations is added after 30 min of reaction, is at war with reference to anti-
Should, to confirm developer and targeted integration whether there is specificity.By 24 orifice plate warm bath different times after sample-adding（0、15、30、
60、90、120min）, the free developer of PBS rinsings, pancreatin digestion, after collecting cell, cell is detected in difference with gamma counter
The intake situation of time developer.Above-mentioned experiment is repeated twice, and multiple holes are set per hole.
The certain density developer of high expression is absorbed in Fig. 1（18F-NOTA-Dimer-San）In the cell of different time
Intake situation, during 90 min developer intake reach peak value, cell Percentage bound is 8.02 ± 2.02%；Low expression is absorbed to add
Cellular uptake declines after certain blocking agent, and cell Percentage bound is reduced to during 90 min：4.45±1.29%.Two groups of data are examined through t,
Significant difference, P be present<0.05, illustrate developer18F-NOTA-Dimer-San is combined with target spot Hsp90 with specificity.
【Biodistribution is tested】
6 nude mices are taken, are randomly divided into 2 groups, pass through tail vein injection developer18F -NOTA-Dimer-San（Every injection
Amount：100-200 μCi）, nude mice is put to death in 3h, separation blood, the heart, liver, pancreas, spleen, kidney, gall-bladder, small intestine, tubule is put into, weighs,
Gamma counter measures radiocounting, and the data obtained calculates %ID/g after correction for attenuation（percentage of injected
Dose per gram of tissue, per g tissue injection dose numbers, for weighing the height of histoorgan increased radioactivity
It is low）, draw bar chart（Referring to Fig. 2）.
Figure it is seen that the developer is mainly drained by digestive systems such as liver, gall-bladder, intestines and stomach, pancreas background
Relative intake is relatively low；Secondly mainly drained by urinary system.
【Imaging】
Model of nude mice bearing tumor and inflammatory model are built first：
When pancreas cancer cell strain PL45 is in increased logarithmic phase, cell is collected in sterile working, and cell is injected in nude mice right shoulder
Number 1 × 107/ only, etc. tumour length to the mm of volume 100 ~ 2003Shi Jinhang Micro-PET are imaged.Leg muscle after on the right side of the nude mice
The μ L of turpentine oil 100 are injected in gap, are built aseptic inflammation model, are imaged after 24 h.
【Tumor bearing nude mice images】
Imaged with Micro-PET toy scanners.Nude mice is imaged with after 2% isoflurane anesthesia by tail vein injection
Agent（18F-NOTA-Dimer-San）100-200 μ Ci/ only, are placed in prone position and imaged respectively at 1 h, 2 h（See Fig. 5）.
18F-NOTA-Dimer San tumor imagings：The visible right side tumor position developer intakes of Fig. 6 are increased.
【Inflammatory model images】
18F-NOTA-Dimer San inflammatory lesions image：The visible inflammation parts of Fig. 7 have no that developer absorbs.
3rd, the present invention is easy to implement, it is not necessary to which special chemistry synthesis module, cost are low." two-step method " is used to improve production
Rate, HPLC（High performance liquid chromatography）As a result show that mark rate reaches 25 ~ 30%.
Brief description of the drawings
The embodiment of the present invention is described in further detail below in conjunction with the accompanying drawings.
Fig. 1 is the present invention18F-NOTA-Dimer San cellular uptakes are tested and blocking experiment result.
Fig. 2 is distribution experiments in normal nude mice body of the present invention.
Fig. 3 is cancer of pancreas model of nude mice bearing tumor of the present invention（Tumor size：0.72*0.6cm）.
Fig. 4 is nude mice aseptic inflammation model of the present invention (obvious see thigh swelling behind the right side).
Fig. 5 is tumor bearing nude mice of the present invention18F-NOTA-Dimer San Micro-PET positive scintigraphies (1 h).
Fig. 6 is tumor bearing nude mice of the present invention18F-NOTA-Dimer San Micro-PET positive scintigraphies (2 h).
Fig. 7 is tumor bearing nude mice of the present invention18F-NOTA-Dimer San Micro-PET MIP scheme (1 h).
Fig. 8 is inflammatory model of the present invention18The negative imagings (1 h, 2 h) of F-NOTA-San Micro-PET.
Fig. 9 is inflammatory model of the present invention18F-NOTA-San Micro-PET MIP scheme (1 h).
Embodiment
The preparation method of the Dimer-San A Cyclopeptide derivatives cancer of pancreas molecular probes of the F-18 of embodiment 1 marks, including
Following steps：
(1) prepare18F- normal saline solutions：
The 50 mCi Na that cyclotron is produced18F solution is first enriched in respectively through the mol/L of 10 mL 0.5 and pH=8.4
NaOAc solution and the Sep-Park light QMA posts of 10 mL deionized waters pretreatment；Then removed with the elution of 5 mL deionized waters
Remove metal impurities ion；Eluted and collected with the mL physiological saline of 0.2 mL ~ 1 again, obtained18F- normal saline solutions.
(2) μ L of acetic acid 5,0.01M AlCl are added in 5mL vials312 μ L's and 10 mCi18F- physiology
The μ L of saline solution 100,10 min are heated at a temperature of 120 DEG C, obtain reaction solution.
(3) the μ g of NOTA-Dimer-Sansalvamide A polypeptides 100 are weighed in 1.5 mL EP pipes, add 250 μ L second
Nitrile, fully dissolve after the min of ultrasonic oscillation 2, add 40 μ L deionized waters, the polypeptide dissolved；The polypeptide of dissolving moves
Into reaction solution, 5 min are cooled down after 10 min are heated at a temperature of 100 DEG C, obtain label18F-NOTA-Dimer-
Sansalvamide A, referred to as18F-NOTA-Dimer-San, HPLC（High performance liquid chromatography）As a result show mark rate for 25 ~
30%, its structural formula is as follows：
。
(4) label18F-NOTA-Dimer-San removes solvents with dry machine is hanged in 40 DEG C, and with the phosphoric acid containing 5% DMSO
Salt buffer（PBS）Dissolving, is most produced through 0.22 μm of membrane filtration afterwards18The Dimer-Sansalvamide A cyclic peptide of F marks
Derivative cancer of pancreas molecular probe.
Wherein：Phosphate buffer and label18F-NOTA-Dimer-San ratio is 300 μ L：100 μg.
The preparation method of the Dimer-San A Cyclopeptide derivatives cancer of pancreas molecular probes of the F-18 of embodiment 2 marks, including
Following steps：
(1) prepare18F- normal saline solutions：
The 100 mCi Na that cyclotron is produced18F solution is first enriched in respectively through the mol/L of 10 mL 0.5 and pH=8.4
NaOAc solution and 10 mL deionized waters pretreatment Sep-Park light QMA posts；Then washed with 5 mL deionizations
Metal impurities ion is removed in removing；Eluted and collected with the mL physiological saline of 0.2 mL ~ 1 again, obtained18F- normal saline solutions.
(2) μ L of acetic acid 5,0.01M AlCl are added in 5 mL vials312 μ L's and 20 mCi18F- physiology
The μ L of saline solution 100,10 min are heated at a temperature of 120 DEG C, obtain reaction solution.
(3) the μ g of NOTA-Dimer-Sansalvamide A polypeptides 150 are weighed in 1.5 mL EP pipes, add 350 μ L second
Nitrile, fully dissolve after the min of ultrasonic oscillation 2, add 40 μ L deionized waters, the polypeptide dissolved；The polypeptide of dissolving moves
Into reaction solution, 5 min are cooled down after 10 min are heated at a temperature of 105 DEG C, obtain label18F-NOTA-Dimer-
Sansalvamide A, referred to as18F-NOTA-Dimer-San, HPLC（High performance liquid chromatography）As a result show mark rate for 25 ~
30%, its structural formula is the same as embodiment 1.
(4) label18F-NOTA-Dimer-San removes solvents with dry machine is hanged in 40 DEG C, and with the phosphoric acid containing 5% DMSO
Salt buffer（PBS）Dissolving, is most produced through 0.22 μm of membrane filtration afterwards18The Dimer-Sansalvamide A cyclic peptide of F marks
Derivative cancer of pancreas molecular probe.
Wherein：Phosphate buffer and label18F-NOTA-Dimer-San ratio is 500 μ L：150 μg.
The preparation method of the Dimer-San A Cyclopeptide derivatives cancer of pancreas molecular probes of the F-18 of embodiment 3 marks, including
Following steps：
(1) prepare18F- normal saline solutions：
The 75 mCi Na that cyclotron is produced18F solution is first enriched in respectively through the mol/L of 10 mL 0.5 and pH=8.4
NaOAc solution and the Sep-Park light QMA posts of 10 mL deionized waters pretreatment；Then removed with the elution of 5 mL deionized waters
Remove metal impurities ion；Eluted and collected with the mL physiological saline of 0.2 mL ~ 1 again, obtained18F- normal saline solutions.
(2) μ L of acetic acid 5,0.01M AlCl are added in 5 mL vials312 μ L's and 15 mCi18F- physiology
The μ L of saline solution 100,10 min are heated at a temperature of 120 DEG C, obtain reaction solution.
(3) the μ g of NOTA-Dimer-Sansalvamide A polypeptides 125 are weighed in 1.5 mL EP pipes, add 300 μ L second
Nitrile, fully dissolve after the min of ultrasonic oscillation 2, add 40 μ L deionized waters, the polypeptide dissolved；The polypeptide of dissolving moves
Into reaction solution, 5 min are cooled down after 10 min are heated at a temperature of 103 DEG C, obtain label18F-NOTA-Dimer-
Sansalvamide A, referred to as18F-NOTA-Dimer-San, HPLC（High performance liquid chromatography）As a result show mark rate for 25 ~
30%, its structural formula is the same as embodiment 1.
(4) label18F-NOTA-Dimer-San removes solvents with dry machine is hanged in 40 DEG C, and with the phosphoric acid containing 5% DMSO
Salt buffer（PBS）Dissolving, is most produced through 0.22 μm of membrane filtration afterwards18The Dimer-Sansalvamide A cyclic peptide of F marks
Derivative cancer of pancreas molecular probe.
Wherein：Phosphate buffer and label18F-NOTA-Dimer-San ratio is 400 μ L：125 μg.
Claims (3)
- The preparation method of the Dimer-San A Cyclopeptide derivatives cancer of pancreas molecular probes of 1.F-18 marks, comprises the following steps：(1) prepare18F- normal saline solutions：50 ~ 100mCi Na that cyclotron is produced18F solution is first enriched in respectively through the mol/L of 10 mL 0.5 and pH=8.4 NaOAc solution and 10 mL deionized waters pretreatment Sep-Park light QMA posts；Then washed with 5 mL deionizations Metal impurities ion is removed in removing；Eluted and collected with the mL physiological saline of 0.2 mL ~ 1 again, obtained18F- normal saline solutions；(2) μ L of acetic acid 5,0.01 M AlCl are added in 5 mL vials312 μ L's and 10 ~ 20 mCi is described18F- The μ L of normal saline solution 100,10 min are heated at a temperature of 120 DEG C, obtain reaction solution；(3) the μ g of NOTA-Dimer-Sansalvamide A polypeptides 100 ~ 150 are weighed in 1.5 mL EP pipes, add 250 ~ 350 μ L acetonitriles, fully dissolve after the min of ultrasonic oscillation 2, add 40 μ L deionized waters, the polypeptide dissolved；The dissolving Polypeptide move in the reaction solution, cool down 5 min after 10 min are heated at a temperature of 100 ~ 105 DEG C, obtain label18F- NOTA-Dimer-Sansalvamide A, referred to as18F-NOTA-Dimer-San, its structural formula are as follows：；(4) the label18F-NOTA-Dimer-San removes solvents with dry machine is hanged in 40 DEG C, and with the phosphoric acid containing 5% DMSO Salt buffer dissolves, and is most produced afterwards through 0.22 μm of membrane filtration18The Dimer-Sansalvamide A Cyclopeptide derivatives of F marks Cancer of pancreas molecular probe.
- 2. the preparation side of the Dimer-San A Cyclopeptide derivatives cancer of pancreas molecular probes of F-18 marks as claimed in claim 1 Method, it is characterised in that：The condition of the step (3) middle supersonic oscillations refers to that temperature is 37 DEG C, and the concussion time is 2 min.
- 3. the preparation side of the Dimer-San A Cyclopeptide derivatives cancer of pancreas molecular probes of F-18 marks as claimed in claim 1 Method, it is characterised in that：The step (4) middle phosphate buffer and the label18F-NOTA-Dimer-San ratio is 300~500 μL：100~150 μg.
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| CN108250276B (en) * | 2018-02-06 | 2022-02-01 | 李玉民 | Preparation method of Cu-64-labeled Hsp90 inhibitor pancreatic cancer diagnosis and treatment integrated molecular probe |
| CN117624278A (en) * | 2024-01-25 | 2024-03-01 | 中国药科大学 | Specific tumor diagnosis probe and imaging agent for targeting heat shock protein 90 |
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