CN106492237B - A kind of isoDGR polypeptide radiopharmaceuticals and its preparation method and application - Google Patents
A kind of isoDGR polypeptide radiopharmaceuticals and its preparation method and application Download PDFInfo
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Abstract
The invention provides a kind of isoDGR polypeptide radiopharmaceuticals and its preparation method and application, the isoDGR polypeptide radiopharmaceuticals include isoDGR polypeptides and radionuclide99mTc, the medicine is by bifunctional chelating agent HYNIC (hydrazinonicotinamide, buzane niacinamide) by radionuclide99mTc is tagged on new isoDGR peptide molecules, and labeled drug gathers tumor locus by the way that the targetings of isoDGR polypeptides is dense in vivo, and using the single photon tomography technology of nuclear medicine, localization diagnosis is carried out to kinds of tumors.
Description
Technical field
The present invention relates to diagnosing tumor radiopharmaceutical, more particularly to a kind of isoDGR polypeptide radiopharmaceuticals and its preparation
Methods and applications.
Background technology
Integrin is a kind of heterodimer transmembrane receptor, is made up of α and β Liang Ge subunits, is had not in composition at least 24
The heterodimer of congenerous and ligand-binding activity.They as two-way signaling molecular regulation cell interior and outside signal from
And regulate and control various important cell functions, such as adhesion, polarity, differentiation, migration and cell division.It is abnormal under pathological state
Integrin signaling cause abnormal cell division, migrate and stick, these are the marks of tumour and its transfer.Wherein, integrate
Plain αvβ6Be unique in that, its single-minded expression is in epithelial cell, and β6Only with αvForm single heterodimer.Integrate
Plain αvβ6In embryo development procedure, height expression developmental lung, skin and kidney epithelial cell, health adult in it
Expression lower.Integrin alphavβ6In cancer of pancreas, breast cancer, lung cancer, oral cavity and cutaneous squamous cell carcinoma（SCC）, colon cancer, stomach
Up-regulated expression in cancer and carcinoma of endometrium.And integrin alpha5β1It is analogous to integrin alphavβ3Important cell adhesion molecule, be
The major receptors of fibronectin, main function is to cause fibronectin extracellular matrix to be assembled, and integrin family is important
Forming member.Recent studies have found that integrin alpha5β1Effect in neonate tumour blood vessel is equally important, is given birth in new vessels
Regulate and control propagation and the migration of endothelial cell during.In addition, integrin alpha5β1Maturation normal cell and blood vessel do not express or
Person's low expression, and the high expression in tumor neogenetic blood vessels, it has now been found that its such as colorectal cancer, breast cancer, oophoroma,
High expression in a variety of tumours such as lung cancer, stomach cancer and neural glioma.
The content of the invention
It is described it is an object of the invention to provide a kind of isoDGR polypeptide radiopharmaceuticals and its preparation method and application
IsoDGR polypeptide radiopharmaceuticals are a kind of new to be used for integrin alpha5β1And αvβ6The polypeptide radiation of positive tumor early diagnosis
Property medicine, the medicine will be radiated by bifunctional chelating agent HYNIC (hydrazinonicotinamide, buzane niacinamide)
Property nucleic99mTc is tagged on new isoDGR peptide molecules, and labeled drug is dense by the targeting of isoDGR polypeptides in vivo
Gather tumor locus, using the single photon tomography technology of nuclear medicine, localization diagnosis is carried out to kinds of tumors.
The purpose of the present invention is to be achieved through the following technical solutions：
A kind of isoDGR polypeptide radiopharmaceuticals, including isoDGR polypeptides and radionuclide99mTc, the isoDGR are more
Peptide is D-PG-special-shaped aspartic-glycine-arginine-D- type arginine five-membered ring peptide sequences, the radioactivity
Nucleic99mTc marks the isoDGR polypeptides by bifunctional chelating agent HYNIC, that is, obtains the isoDGR polypeptides radioactivity medicine
Thing ---99mTc-HisoDGR, the isoDGR polypeptide radiopharmaceuticals are colourless transparent liquid injection.
The preparation method of described isoDGR polypeptide radiopharmaceuticals, comprises the following steps：
1）HYNIC-c (phg-isoDGRk) preparation
C (phg-isoDGRk) is dissolved in the mixed liquor that pH is 9.0 and obtains mixed liquor A, the mixed liquor is 0.1 M
NaHCO3With 0.1 M Na2CO3By 9:1 mixing gained, then HYNIC-NHS is dissolved in DMF and obtains mixed liquid B；By mixed liquor A
Mixed with mixed liquid B, be placed on 30 DEG C of oscillators and react 24 hours, product is through YMC-Pack ODS-A C18 semi-preparative columns
HPLC is isolated and purified, and collects the cut of object, is merged collection liquid and is freezed, that is, obtains the HYNIC-c (phg-
isoDGRk)；
2）99mTc-HYNIC- c (phg-isoDGRk) preparation
Prepare sodium trisulfonate containing triphenylphosphine, trihydroxy methyl glycine, disodium succinate, butanedioic acid and HYNIC- c
(phg-isoDGRk) the mL of mixed liquor 500 adds 1.0-1.5 mL Na in 10 mL cillin bottles99mTcO4Solution, 100
DEG C heating water bath cillin bottle reaction 20-25 minutes, question response terminate rear room temperature and cooled down 10 minutes, that is, the radiation of isoDGR polypeptides is made
Property medicine, through radioactivity HPLC Analysis of quality control top coal drawings.
Further, the step 2）Described in sodium trisulfonate containing triphenylphosphine, trihydroxy methyl glycine, butanedioic acid two
In the mixed liquor of sodium, butanedioic acid and HYNIC- c (phg-isoDGRk), the content of each material is：
The mg of triphenylphosphine sodium trisulfonate 5.0
The mg of trihydroxy methyl glycine 6.5
The mg of disodium succinate 38.5
The mg of butanedioic acid 12.7
HYNIC- c(phg-isoDGRk) 20 μg。
Further, step 1）The semi-preparative column HPLC isolation and purification methods are：Use the Infinity of Agilent 1260
HPLC system is equipped with YMC-Pack ODS-A C18 semi-preparative columns, gradient elution 35 minutes, the wherein mL/min of flow velocity 4, mobile phase
A is the deionized water containing 0.05% TFA, and B is the acetonitrile containing 0.05% TFA, and elution gradient is set as 100% A and 0% during starting
B, 50% A and 50% B at 25 minutes, 0% A and 100% B at 30 minutes, 100% A and 0% B at 35 minutes.
Further, step 1）The semi-preparative column HPLC isolation and purification methods are：Use the Infinity of Agilent 1260
HPLC system is equipped with YMC-Pack ODS-A C18 semi-preparative columns（250 mm × 10 mm, 5 μm, 12 nm）, gradient elution
35 minutes, the mL/min of flow velocity 4, wherein mobile phase A were the deionized water containing 0.05% TFA, and B is the acetonitrile containing 0.05% TFA,
Elution gradient is set as 100% A and 0% B during starting, 50% A and 50% B at 25 minutes, 0% A and 100% B at 30 minutes, 35
100% A and 0% B during minute.
Further, step 2）The method of the radioactivity HPLC Analysis of quality control is：Radioactivity HPLC Quality Controls use
The Infinity HPLC operating systems of Angilent Techologies 1260, equipped with Raytest Gabi radioactive detectors,
And Agilent Zorbax SB-C18 column（4.6 mm × 250 mm, 5 μm）, mobile phase A is 0.05% TFA water
Solution, Mobile phase B is 0.05% TFA acetonitriles, right99mTc-HYNIC-isoDGR4# carries out gradient elution 30 minutes, flow velocity 1.0
ML/min, elution gradient are set as 100% A and 0% B during starting, keep at 5 minutes start gradient constant, 90% A at 10 minutes
With 10% B, 40% A and 60% B at 20 minutes, the A of baseline gradient 100% and 0% B are returned at 30 minutes.
The application of described isoDGR polypeptide radiopharmaceuticals, the isoDGR polypeptide radiopharmaceuticals are whole available for preparing
Close plain avb6With integrin a5b1Double positive and integrin avb6Or integrin a5b1The radiation of single positive tumor imaging diagnosis
Property medicine.
Further, the integrin avb6With integrin a5b1Double positive and integrin avb6Or integrin a5b1Dan Yang
Property tumour include colorectal cancer, breast cancer, oophoroma, lung cancer, stomach cancer, glioma, cancer of pancreas, colon cancer, lung cancer.
The isoDGR polypeptides are c (phg-isoDGRk) (cyclo (D-ph-Gly- isoAsp-Gly-Arg-D-Lys)
(c represents cyclic peptide, and phg represents D types-phenylglycine, and isoD represents special-shaped aspartic acid, and G represents glycine, and R represents smart ammonia
Acid, k represent D types-arginine) (Fig. 1-upper left).Design in the present invention99mTc-HisoDGR molecular probes are first by double work(
Energy chelating agent HYNIC is connected with isoDGR polypeptides, is then carried out in the presence of synergy modes99mTc is marked, that is, is synthesized99mTc-
HisoDGR (Fig. 1-right side).
The present invention having the beneficial effect that compared with prior art：
1st, use HYNIC in the present invention as bifunctional chelating agent, at the same use tricine (trihydroxy methyl glycine) and
TPPTS (trisodium triphenylphosphine- 3,3', 3''-trisulfonate, triphenylphosphine sodium trisulfonate)
As synergy modes so that "99mTc-HYNIC cores " have external stability inside more good；
2nd, different aspartic-glycine-arginine in the present invention（isoDGR）Sequence is to be different from RGD（Arginine-sweet ammonia
Acid-aspartic acid）The new amino acid sequence of the targeted integration element family of sequence, new isoDGR five-membered rings peptide c (phg-
IsoDGRk it is) by simulating c (X- respectively on molecular docking binding model on tri- amino acid sequences of isoDGR
) and integrin alpha isoDGR-X5β1、αvβ6And αvβ3With reference to conformation, screen the obtained part of high-affinity.Result of study table
It is bright, c (phg-isoDGRk) polypeptide energy and integrin alpha5β1And αvβ6High-affinity combines, the radioactivity based on the sequences Design point
Sub- probe can be effectively used to integrin alpha5β1And αvβ6The diagnosis of positive tumor.
Brief description of the drawings
Fig. 1 is isoDGR polypeptides and HYNIC conjugate（A）And its99mTc tag structure schematic diagrames（B）With HPLC（It is high
Effect liquid phase chromatogram）Spectrogram（C）；
Fig. 2 is Flow cytometry BxPC-3, U87MG and HT-29 integrin（Integrin）αvβ6And α5β1Expression
Situation,（A-B）BxPC-3 cells are integrin alphavβ6And α5β1Double positive expressions,（C-D）U87MG cells are integrin alpha5β1Dan Yang
Property expression,（E-F）HT-29 cells are integrin alphavβ6Single positive expression；
Fig. 3 is99mTc-HisoDGR is in integrin alpha5β1And αvβ6Double positive Bx-PC3 human pancreas cancers tumor models 0.5,1
With 2 h (A) and the h of control group enclosed experiment 0.5 bio distribution result (B)；
Fig. 4 is integrin alpha5β1And αvβ6Double positive BxPC-3 human pancreas cancers tumor model injections99mTc-HisoDGR（A）With
And the control group closed with excessive cold peptide（B）0.5, the 1 and 2 h nanoSPECT/CT imaging figures after injection；
Fig. 5 is integrin alpha5β1Positive U87MG human gliomas nude mice injection99m0.5,1 and 2 h after Tc-isoDGR（A）With
And the h of control group 0.5 closed with excessive cold peptide（B）NanoSPECT/CT imaging figures, arrow represents tumor locus；
Fig. 6 is integrin alphavβ6Positive HT-29 human colon carcinomas tumour nude mice injection99m0.5,1 and 2 h after Tc-isoDGR（A）
And the h of control group 0.5 closed with excessive cold peptide（B）NanoSPECT/CT imaging figure.
Embodiment
Below in conjunction with drawings and Examples, the invention will be further described.
1. material
Dicyclcohexylcarbodiimide (DCC, N, N'- dicyclohexylcarbodiimide), N-
Hydroxysuccinimide (NHS, n-hydroxysuccinimide), succinic acid (butanedioic acid), trisodium
Triphenylphosphine-3,3', 3''-trisulfonate (TPPTS, triphenylphosphine sodium trisulfonate), tricine (three
Hydroxymethylglycinate) it is purchased from Sigma-Aldrich.HYNIC-NHS is purchased from Noca-biochem companies of the U.S..
(cyclo (D-ph-Gly- isoAsp-Gly-Arg-D-Lys) commissions are uncommon to apply biological section to isoDGR polypeptides c (phg-isoDGRk)
Skill (Shanghai) Co., Ltd. (U.S. is uncommon to apply biological (CSBio) overseas first branch company) order synthesis.Na99mTcO4Eluent
Purchased from Beijing Atom High Tech Co., Ltd.
2 methods and result
2.1 HPLC methods
HPLC methods 1：YMC-Pack ODS-A C18 analyses are equipped with using the Infinity HPLC systems of Agilent 1260
Post（250 mm × 4.6 mm, 5 μm, 12 nm）, gradient elution 35 minutes, the mL/min of flow velocity 1, wherein mobile phase A for go from
Sub- water（Containing 0.05% TFA）, B is acetonitrile（Containing 0.05% TFA）.100% A and 0% B when elution gradient is set as originating, 25 points
50% A and 50% B during clock, 0% A and 100% B at 30 minutes, 100% A and 0% B at 35 minutes.
HPLC methods 2：YMC-Pack ODS-A C18 half are equipped with using the Infinity HPLC systems of Agilent 1260 to make
Standby post（250 mm × 10 mm, 5 μm, 12 nm）, gradient elution 35 minutes, the mL/min of flow velocity 4, wherein mobile phase A are to go
Ionized water（Containing 0.05% TFA）, B is acetonitrile（Containing 0.05% TFA）.100% A and 0% B when elution gradient is set as originating, 25
Minute when 50% A and 50% B, 0% A and 100% B at 30 minutes, 100% A and 0% B at 35 minutes.
HPLC methods 3：Radioactivity HPLC Quality Controls are grasped using the Infinity HPLC of Angilent Techologies 1260
Make system, equipped with Raytest Gabi radioactive detectors, and Agilent Zorbax SB-C18 column（4.6 mm
× 250 mm, 5 μm）.Mobile phase A is the 0.05% TFA aqueous solution, and Mobile phase B is 0.05% TFA acetonitriles, right99mTc-
HYNIC-isoDGR4# carry out gradient elution 30 minutes, the mL/min of flow velocity 1.0, elution gradient be set as starting when 100% A and
0% B, keep at 5 minutes start gradient constant, 90% A and 10% B at 10 minutes, 40% A and 60% B at 20 minutes, 30 minutes
When return to the A of baseline gradient 100% and 0% B.
2.2 HYNIC-c (phg-isoDGRk) (HisoDGR) preparation
Accurately weigh the mg of polypeptide isoDGR polypeptides 4.25（7.3 μmol）, 600 μ L pH are dissolved in for 9.0（0.1 M
NaHCO3With 0.1 M Na2CO3By 9:1 mixing）The aqueous solution in, then weigh the mg of HYNIC-NHS 6.4（14.5 μmol）, it is molten
In 100 μ L DMF.The solution of two kinds of reactants is mixed, in light yellow transparent liquid.Place reaction liquid into 30 DEG C of vibrations
Reacted 24 hours on device.Pass through HPLC in course of reaction（YMC-Pack ODS-A C18 analytical columns, HPLC methods 1）Detection reaction
Process.After reactant polypeptide isoDGR polypeptides reactives are complete, by product HPLC（It is prepared by YMC-Pack ODS-A C18 half
Post, HPLC methods 2）Isolated and purified, collect the product for merging the min of 2 retention time of HPLC methods 21.5.After freeze-drying
Obtain the mg of white powdery solids product HisoDGR 5.Product tests and analyzes purity through HPLC analytical columns, and purity is more than 95%.
Confirmed again through MALDI-TOF mass spectral analyses, mass spectrometry results: m/z =893.19 ( [M+H]+), C39H49N12O11S]+Reason
It is 893.33 by value.
2.3 99mTc-HisoDGR preparation
Prepare the mg containing TPPTS 5.0 mg, tricine 6.5, disodium succinate（disodium succinate
hexahydrate）38.5mg, the mg of butanedioic acid 12.7 and 20 μ g HisoDGR（1 μ g/ μ L, are dissolved in ultra-pure water）Mixing
The mL of liquid 500 adds 1.0-1.5 mL Na in 10 mL cillin bottles99mTcO4Solution (10-50 mCi), 100 DEG C of water-baths
Heat cillin bottle to react 20-25 minutes, question response terminates rear room temperature and cooled down 10 minutes, samples and analyzes in radioactivity HPLC
(HPLC methods 3),99mTc-HisoDGR mark rates>95%, through Sep-Pak C18 posts radiochemical purity after purification>98%.
2.4 99mTc-HisoDGR is in tumor bearing nude mice bio distribution
Fig. 2 is Flow cytometry BxPC-3, U87MG and HT-29 integrin（Integrin）αvβ6And α5β1Expression
Situation.As a result it is integrin alpha to show BxPC-3 cellsvβ6And α5β1Double positive expressions（Fig. 2A-B）, U87MG cells are integrin alpha5
β1Single positive expression（Fig. 2 C-D）, HT-29 cells are integrin alphavβ6Single positive expression（Fig. 2 E-F）.
BALB/c lotus BxPC-3 human pancreas cancer tumour nude mices are randomly divided into some groups, every group 4.Each group experiment nude mice point
Not through the mL's of tail vein injection 100 (~ 74 kBq)99mTc mark HisoDGR polypeptides, after injection 30 minutes, 60 minutes and
Nude mice is extremely tested by component other places within 120 minutes, take blood and main organs, weigh and measure radiocounting, after decay correction
Calculate per gram of tissue percentage injection dose rate (%ID/g).Receptor blockade control group is being injected99mCo-injection during Tc-HisoDGR
The excessive cold peptides of isoDGR (~ 400 μ g/ are only), are carried out for 30 minutes, 60 minutes and 120 minutes after injection by above same method
Biodistribution experiments.
In lotus BxPC-3 human pancreas cancer tumour nude mice models, within the observed time99mTc-HisoDGR tumour is taken the photograph
The intake for being only below kidney is taken, almost higher than all other internal organs tumor uptake (Fig. 3 A), the display injection of receptor blockade experimental result
The intake of 0.5 h tumours receives conspicuousness suppression (Fig. 3 B), this explanation afterwards99mTc-HisoDGR is that acceptor is situated between in the intake of tumour
The specificity intake led.
2.4 99mTc-HisoDGR images in the nanoSPECT/CT of tumor bearing nude mice
BALB/c lotus BxPC-3 human pancreas cancers, U87MG human gliomas, HT-29 human colon carcinoma tumour nude mices are divided at random
Into some groups, every group 4.Each group tests nude mice respectively through the mL's of tail vein injection 100 (~ 37 MBq)99mTc-HisoDGR,
Imaged respectively in nanoSPECT/CT imaging systems by group within 30 minutes, 60 minutes and 120 minutes after injection.Acceptor seals
Control group is closed to inject99mThe cold peptides of co-injection excess isoDGR (~ 400 μ g/ are only) during Tc-HisoDGR, 30 points after injection
Clock, 60 minutes and 120 minutes carry out imaging experiment by above same method.
Fig. 4 is shown99mTc-HisoDGR clearly can image to high-contrast BxPC-3 human pancreas cancer tumours, imaging knot
For fruit in addition to kidney has higher intake, other internal organs backgrounds are relatively low.Receptor blockade experiment image results show that tumor uptake can be had
Effect suppresses, and it is the intake of receptor-specific mediation to illustrate tumor imaging.Also indicate that99mTc-HisoDGR, which has, is used for integrin alpha5β1
And αvβ6The application potential of double positive tumor early diagnosis.Meanwhile inject99m30 minutes, 60 minutes and 120 points after Tc-HisoDGR
Clock integrin alpha5β1Positive glioma（U87MG）Tumour and integrin alphavβ6Positive human colon carcinoma（HT29）NanoSPECT/
CT imaging figures（Fig. 5 and Fig. 6）Also can the clear imaging tumor of high-contrast, and receptor blockade experiment in tumor uptake show it is bright
It is aobvious to suppress, show99mTc-HisoDGR is in integrin alpha5β1Or αvβ6The imaging of single positive tumour is all receptor-specific mediation
's.99mTc-HisoDGR, which has, is used for integrin alpha5β1Or αvβ6The application potential of single positive early diagnosis of tumor.
In summary, based on new isoDGR polypeptides radiolabeled probe99mTc-HisoDGR, which has, is used for integrin alpha5
β1And αvβ6Double positive and integrin alphas5β1Or αvβ6The application potential of single positive kinds of tumors early diagnosis.
Claims (8)
1. a kind of isoDGR polypeptide radiopharmaceuticals, including isoDGR polypeptides and radionuclide99mTc, it is characterised in that：It is described
IsoDGR polypeptides are D-PG-special-shaped aspartic-glycine-arginine-D- type arginine five-membered ring peptide sequences, institute
State radionuclide99mTc marks the isoDGR polypeptides, the isoDGR polypeptides radioactivity by bifunctional chelating agent HYNIC
Medicine is colourless transparent liquid injection.
A kind of 2. preparation method of isoDGR polypeptide radiopharmaceuticals as claimed in claim 1, it is characterised in that：The preparation
Method comprises the following steps：
1）HYNIC-c (phg-isoDGRk) preparation
IsoDGR polypeptides are dissolved in the mixed liquor that pH is 9.0 and obtain mixed liquor A, the mixed liquor is 0.1 M NaHCO3With 0.1
M Na2CO3By 9:1 mixing gained, then HYNIC-NHS is dissolved in DMF and obtains mixed liquid B；Mixed liquor A and mixed liquid B are mixed
Close, be placed on 30 DEG C of oscillators and react 24 hours, reaction process is detected by HPLC in course of reaction, as reactant isoDGR
After polypeptides reactive is complete, product isolates and purifies through YMC-Pack ODS-A C18 semi-preparative columns HPLC, collects evaporating for object
Point, merge collection liquid and freeze, that is, obtain the HYNIC-c (phg-isoDGRk)；
2）99mTc-HYNIC- c (phg-isoDGRk) preparation
Prepare sodium trisulfonate containing triphenylphosphine, trihydroxy methyl glycine, disodium succinate, butanedioic acid and HYNIC- c (phg-
IsoDGRk the mL of mixed liquor 500) adds 1.0-1.5 mL Na in 10 mL cillin bottles99mTcO4Solution, 100 DEG C of water-baths
Heat cillin bottle reaction 20-25 minutes, question response terminates rear room temperature and cooled down 10 minutes, that is, isoDGR polypeptide radioactivity medicines are made
Thing, through radioactivity HPLC Analysis of quality control top coal drawings.
3. the preparation method of isoDGR polypeptide radiopharmaceuticals according to claim 2, it is characterised in that：The step 2）
Described in sodium trisulfonate containing triphenylphosphine, trihydroxy methyl glycine, disodium succinate, butanedioic acid and HYNIC- c (phg-
IsoDGRk in mixed liquor), the content of each material is：
The mg of triphenylphosphine sodium trisulfonate 5.0
The mg of trihydroxy methyl glycine 6.5
The mg of disodium succinate 38.5
The mg of butanedioic acid 12.7
HYNIC- c(phg-isoDGRk) 20 μg。
4. the preparation method of isoDGR polypeptide radiopharmaceuticals according to claim 2, it is characterised in that：Step 1）It is described
Detecting the method reacted by HPLC is：YMC-Pack ODS-A are equipped with using the Infinity HPLC systems of Agilent 1260
C18 analytical columns, gradient elution 35 minutes, the wherein mL/min of flow velocity 1, mobile phase A are the deionized water containing 0.05% TFA, and B is
Acetonitrile containing 0.05% TFA, elution gradient are set as 100% A and 0% B during starting, 50% A and 50% B at 25 minutes, 30 points
0% A and 100% B during clock, 100% A and 0% B at 35 minutes.
5. the preparation method of isoDGR polypeptide radiopharmaceuticals according to claim 2, it is characterised in that：Step 1）It is described
Semi-preparative column HPLC isolation and purification methods are：YMC-Pack ODS-A are equipped with using the Infinity HPLC systems of Agilent 1260
C18 semi-preparative columns, gradient elution 35 minutes, the wherein mL/min of flow velocity 4, mobile phase A are the deionized water containing 0.05% TFA, B
For the acetonitrile containing 0.05% TFA, elution gradient is set as 100% A and 0% B during starting, 50% A and 50% B at 25 minutes, 30
Minute when 0% A and 100% B, 100% A and 0% B at 35 minutes.
6. the preparation method of isoDGR polypeptide radiopharmaceuticals according to claim 2, it is characterised in that：Step 2）It is described
The method of radioactivity HPLC Analysis of quality control is：Radioactivity HPLC Quality Controls use Angilent Techologies 1260
Infinity HPLC operating systems, equipped with Raytest Gabi radioactive detectors, and Agilent Zorbax SB-C18
Column, mobile phase A are the 0.05% TFA aqueous solution, and Mobile phase B is 0.05% TFA acetonitriles, right99mTc-HYNIC-isoDGR4#
Gradient elution 30 minutes, the mL/min of flow velocity 1.0 is carried out, elution gradient is set as 100% A and 0% B during starting, protected at 5 minutes
It is constant to hold start gradient, 90% A and 10% B at 10 minutes, 40% A and 60% B at 20 minutes, returns to baseline gradient at 30 minutes
100% A and 0% B.
A kind of 7. application of isoDGR polypeptide radiopharmaceuticals as claimed in claim 1, it is characterised in that：The isoDGR is more
Peptide radiopharmaceutical is used to prepare integrin avb6With integrin a5b1Double positive or integrin avb6Or integrin a5b1Dan Yang
Property tumor imaging diagnosis radiopharmaceutical.
8. the application of isoDGR polypeptide radiopharmaceuticals according to claim 7, it is characterised in that：The integrin avb6
With integrin a5b1Double positive or integrin avb6Or integrin a5b1Single positive tumour includes colorectal cancer, breast cancer, ovum
Nest cancer, lung cancer, stomach cancer, glioma, cancer of pancreas, colon cancer, lung cancer.
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