CN110305187A - Prostate cancer PET diagnostic reagent68Ga-NOTA-ANCP-PSMA and its preparation method and application - Google Patents
Prostate cancer PET diagnostic reagent68Ga-NOTA-ANCP-PSMA and its preparation method and application Download PDFInfo
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Abstract
The invention discloses a kind of prostate cancer PET diagnostic reagents68Ga-NOTA-ANCP-PSMA and its preparation method and application.It is described68The structural formula of Ga-NOTA-ANCP-PSMA is as follows:The preparation method includes the following steps: that (S1) synthesizes precursor compound NOTA-ANCP-PSMA using solid-phase synthesis；(S2) it is carried out using direct labelling method68Ga radioactive label.It is described68Ga-NOTA-ANCP-PSMA has many advantages, such as that signature velocity is fast, mark rate is high, while having good stability, preferable hydrophily, high sensitivity and high specific.In addition,68Ga-NOTA-ANCP-PSMA is removed comparatively fast in blood, and tumor uptake is higher.
Description
Technical field
The invention belongs to field of medicaments, and in particular to a kind of prostate cancer PET diagnostic reagent68Ga-NOTA-ANCP-PSMA
And its preparation method and application.
Background technique
Prostate cancer is the most common malignant tumour of male, by 2018, annual new cases 42.75 in global range
Ten thousand people reach 36.18 ten thousand people because suffering from prostate cancer death toll every year, are disease incidence second in male cancer, the death rate the 5th
High cancer.Prostate-specific antigen (PSA) screening can provide early diagnosis for most of patient, but for will appear height
The some patientss of risk or transfer stove lead to not provide accurate diagnosis.Early stage accurately diagnoses and has by stages for selection
The treatment of effect is most important.There are certain for traditional imaging mode such as computed tomography (CT) and nuclear magnetic resonance (MRI)
Defect is failed to pinpoint a disease in diagnosis higher with misdiagnosis rate especially in the case where low PSA is horizontal.Using positron emission computerized tomography (PET) for diagnosis
Prostate cancer has extraordinary prospect, using conventional diagnostic reagent, as choline (11C/18) or fluorodeoxyglucose F-Choline
(18F-FDG) carrying out positron emission computerized tomography (PET) has a good effect for the prostate cancer of middle and advanced stage, however for
There are still certain limitations for the prostate cancer and its metastasis (metastases) of early stage.
Prostate-specific membrane antigen (PSMA) is II type transmembrane glycoprotein, is most early in human prostate cancer cells
It is found in LNCaP, PSMA is significantly increased in prostate gland cancer cell, can be used as the promising target of diagnosing and treating.Glutamic acid-
Urea and the like (Glu-Urea-R) is the micromolecular inhibitor for targeting PSMA, especially contains glutamic acid-urea-lysine sequence
Micromolecular inhibitor, can efficiently, specifically in conjunction with PSMA.Glu-Urea-R has biological activity stabilization, follows in vivo
The good feature of ring half-life short, tissue permeability has better application value in terms of molecular imaging in prostate cancer diagnosis.
Use positron radionuclide68Ga label PSMA micromolecular inhibitor can pass through PET-CT in conjunction with prostate gland cancer cell specificity
Imaging is significant to the diagnosis of prostate cancer and its transfer stove,68Ga-PSMA has become the hot spot of international research.Currently,
It pays close attention in the world more68The PSMA diagnostic reagent of Ga label mainly has68Ga-PSMA-11(68Ga-HBED-CC(Ahx)-
PSMA) and68Ga-PSMA-617 has had more clinical research at present, shows in the diagnosis of prostate cancer and its transfer stove
Highly sensitive and specificity, achieves good effect.
PSMA-11 clinically has more application, and is not achieved preferable effect by patent protection, but suitable
It shares in diagnosis, is not suitable for targeted therapy.PSMA-617 has been achieved in terms of the Clinics and Practices of prostate cancer centainly
Progress, but by patent protection.It develops68Diagnosis of the PSMA small molecule tags object of Ga label to prostate cancer and its transfer stove
It is of great significance by stages, screens the integrated PSMA small molecule tags object pair of diagnosis and treatment with researching value and development prospect
The diagnosis and treatment of prostate cancer provide important foundation.
Micromolecular inhibitor of the Glu-Urea-Lys as PSMA target spot is inhibited using it as core design new small molecule
Agent can efficiently, quickly be combined with PSMA；On the basis of PSMA-617, increase the length that phenylalanine extends side chain, fits
When raising lipophilicity, and increase phenylalanine can be improved as side chain and the combination of plasma protein, extends tumour and persistently accumulates
The tired time lays the foundation for the development of diagnosis and treatment integration marker；NOTA is common chelation group, with Ga3+What is chelated is steady
Permanent several Ki reach 30.98, Ke Yiyu68Ga3+Form stable marker.
Summary of the invention
The problem of for background technique, novel has preferable internal body the purpose of the present invention is to provide a kind of
Outer property68The PSMA small molecule compound of Ga label, and its lipophilicity is suitably promoted, it obtains highly sensitive and high specific
PSMA probe is devised with glutamic acid-urea-lysine (Glu-Urea-Lys) for core group, with 1- naphthylalanine, 4- amine
Methyl cyclohexane formic acid, phenylalanine (Ala (Nap)-Cyh-Phe, hereinafter referred to as ANCP) are the PSMA micromolecular inhibitor of side chain68Ga-NOTA-ANCP-PSMA。
To achieve the goals above, the specific technical solution of the present invention is as follows:
A kind of prostate cancer PET diagnostic reagent68Ga-NOTA-ANCP-PSMA, it is described68The tool of Ga-NOTA-ANCP-PSMA
Body structural formula is as follows:
A kind of compound N OTA-ANCP-PSMA, the concrete structure formula of the NOTA-ANCP-PSMA are as follows:
A second object of the present invention is to provide prostate cancer PET diagnostic reagents as described above68Ga-NOTA-ANCP-
The preparation method of PSMA, which is characterized in that described method includes following steps:
(S1) precursor compound NOTA-ANCP-PSMA is synthesized using solid-phase synthesis；
The preparation of compound 3:
Weighing degree of substitution is 0.3mmol/g starting material Fmoc-Lys (Dde)-Wang Resin3g, is added to reactor
In, DMF is added, impregnates 30min；DMF is drained later, adds the 20%Pip/DMF of 3 times of volumes, nitrogen charging gas shielded, to remove
Fmoc reacts 30min, drains 20%Pip/DMF, and DMF is washed 5 times, ninhydrin detection display navy blue；N is put in proportion into,
N'- succinimidyl carbonate DSC, n,N-diisopropylethylamine DIPEA and 4-dimethylaminopyridine DMAP are added appropriate
DMF reacts 1h under nitrogen protection；H-Glu (OtBu)-OtBuHCl is added into reactor, appropriate DMF is added,
Under nitrogen protection, 1h is reacted, obtains compound 3；
The preparation of compound 4:
By the cutting liquid E liquid cleavage reaction 1h of compound 3, determine whether to be fully connected using ninhydrin detection method, if not
It is fully connected, is fed intake again until being fully connected；After being fully connected, 3 times of 2% hydrazine hydrates of volume, DMF solutions are added, react
30min sloughs Dde protection, and DMF is washed 5 times, obtains compound 4；
The preparation of compound 5:
Amino acid, compound 4, benzotriazole-N, N, N needed for side chain is added according to polypeptide solid-state reaction method ', N '-
Tetramethyl isourea hexafluorophosphate HBTU, N- methylmorpholine NMM；It is sequentially connected 1- naphthylalanine, 4- according to the method described above
The amino acid of 3 equivalents is added in amine methyl cyclohexane formic acid, phenylalanine, and appropriate DMF is added, and reacts 1h；Synthetic product is transferred to
It cuts in pipe, E liquid is added, shake cleavage reaction 1h；Cutting liquid is collected into centrifuge tube, 6 times of volume ice ether, low speed is added
Centrifuge precipitating；The crude product of precipitating is washed three times with ether, obtains the crude product of precursor compound 5；Pass through half preparation chromatographic column
Purifying obtains compound 5；
The preparation of compound 6:
On the basis of compound 5, the chelating agent NOTA of 3 equivalents is added, appropriate DMF is added, reacts 1h；By synthetic product
It is transferred in cutting pipe, E liquid is added, shake cleavage reaction 1h；Cutting liquid is collected into centrifuge tube, 6 times of volume ice second are added
Ether, low speed centrifuge precipitating；The crude product of precipitating is washed three times with ether；Compound 6 is obtained by partly preparing chromatography；
The compound 6 is precursor compound NOTA-ANCP-PSMA；
(S2) it is carried out using direct labelling method68Ga radioactive label；
It opens reactor and temperature is set in 95 DEG C；The compound 6NOTA-ANCP-PSMA lyophilized preparation of freezing is taken out,
It is configured to 5mg/mL solution with ultrapure water, 4 μ L is taken to be placed in 1.5mL EP reaction tube, 90 μ L 1.0mol/ are added into reaction tube
LHEPES buffer solution；3.5mL 0.05mol/L HCl solution is taken, is eluted68Ge-68Ga generator；1.0mL is taken to elute68Ga
Reaction tube is added in 0.05mol/L HCl solution, about 111-185MBq, is placed in 95 DEG C of heating module；Under 200rpm revolving speed
React 15min；Solution after reaction is taken out and measures total activity with activity meter, pH value is measured, is then injected into activation
In Sep-pak C18 column, with 3.0mL pure water rinsing C18 column, to clean impurity；0.22 μm of miillpore filter is installed on a cl 8 column,
It is eluted, is collected into Sterile vacuum bottle with the ethanol solution of 1.0mL 95%；When for zoopery, by 95% ethanol rinse
Group be placed on heating module, to be dried with nitrogen, redissolved with physiological saline, obtain target label object68Ga-NOTA-ANCP-
PSMA；
Synthetic route are as follows:
As a preferred solution, in the preparation of the compound 3, Fmoc-Lys (Dde)-Wang Resin,
The molar ratio of DSC, DIPEA, DMAP are 1: 6: 12:1；
Fmoc-Lys (Dde)-Wang Resin, H-Glu (OtBu)-OtBuHCl, DIPEA, DMAP molar ratio be 1:
3:9:1.
As a preferred solution, in the preparation of compound 4, pure TFA in the cutting liquid E liquid, thioanisole,
Water, phenol, 1,2- dithioglycol mol ratio be 35:2:1:1:1.
As a preferred solution, in the preparation of the compound 5, compound 4, amino acid, benzotriazole-N,
N, N ', N '-tetramethyl isourea hexafluorophosphate, N- methylmorpholine molar ratio be 1:3:2.85:6.
Third object of the present invention is to provide a kind of prostate cancer PET diagnostic reagents as described above68Ga-NOTA-
ANCP-PSMA in prostate cancer diagnosis, by stages with the application in therapeutic evaluation.
The beneficial effects of the present invention are:
68Ga label68Ga-NOTA-ANCP-PSMA has the characteristics that signature velocity is fast, mark rate is high, completes within 15 minutes
Label and purifying, top coal drawing are greater than 95%.Stability is good, and top coal drawing remains at 95% or so in 2 hours, compound
There is preferable hydrophily.68Ga-NOTA-ANCP-PSMA mainly passes through kidney excretion, and other non-target organ intakes are lower, swollen
There is certain intake in tumor.In the diagnosis research of PET-CT, preferable sensitivity and specificity are shown, have in knub position
Clearly image, other than kidney and bladder, other organs and tissue intake are lower.68Ga-NOTA-ANCP-PSMA have compared with
Good inside and outside property is expected to become novel prostatic cancer specific targeting diagnosis reagent.
Detailed description of the invention
Fig. 1 is the mass spectrogram of precursor compound NOTA-ANCP-PSMA；
The HPLC UV map of Fig. 2 precursor compound NOTA-ANCP-PSMA；
Fig. 3 is after purification68The HPLC AD map of Ga-NOTA-ANCP-PSMA；
Fig. 4 is in PBS and BSA68The stability of Ga-NOTA-ANCP-PSMA；
Fig. 5 is68The intracorporal PET-CT imaging of Ga-NOTA-ANCP-PSMA tumor bearing nude mice at 60 minutes.
Specific embodiment
For the ease of further appreciating that examples provided below has done more detailed description to it to of the invention.This
A little embodiments are only not supposed to be a limitation to the present invention for narration or implementation principle, and protection scope of the present invention is still with right
Subject to it is required that.
Embodiment 1
The present embodiment provides a kind of prostate cancer PET diagnostic reagents68The preparation method of Ga-NOTA-ANCP-PSMA, it is described
Method includes the following steps:
(S1) precursor compound NOTA-ANCP-PSMA is synthesized using solid-phase synthesis；
The preparation of compound 3
Synthetic route is as follows, and with Fmoc-Lys (Dde)-Wang Resin (compound 1) for starting material, weighing is taken
Dai Du is the 3g of 0.3mmol/g raw material, is added into reactor, and DMF is added, and impregnates 30min.DMF is drained later, adds 3 times of bodies
Long-pending 20%Pip/DMF, nitrogen charging gas shielded, to remove Fmoc, reaction 30min drains 20%Pip/DMF, and DMF is washed 5 times,
Ninhydrin detection display navy blue.It is put in proportion into N, N'- succinimidyl carbonate (DSC), N, N- diisopropylethylamine
(DIPEA) and 4-dimethylaminopyridine (DMAP), investment ratio are resin: DSC:DIPEA: DMAP=1: 6: 12:1, it is added suitable
DMF is measured, under nitrogen protection, reacts 1h.H-Glu (OtBu)-OtBuHCl (compound 2) is added into reactor, investment
Ratio is compound 1: appropriate DMF is added in compound 2:DIPEA:DMAP=1:3:9:1, under nitrogen protection, reacts 1h, obtains
Obtain compound 3.
The preparation of compound 4
By compound 3, with cutting liquid E liquid, (mol ratio is pure TFA: thioanisole: water: phenol: EDT (1,2- ethylene dithiol
Alcohol)=35:2:1:1:1) cleavage reaction 1h, determine whether to be fully connected using ninhydrin detection method, if not being fully connected, again
It feeds intake until being fully connected.After being fully connected, 3 times of 2% hydrazine hydrates of volume, DMF solutions are added, (15min changes one to reaction 30min
Not good liquor), Dde protection is sloughed, DMF is washed 5 times, obtains compound 4.
The preparation of compound 5
Amino acid and other reagents needed for side chain is added according to polypeptide solid-state reaction method are added later, amino acid, which feeds intake, to rub
That ratio are as follows: compound 4: amino acid: HBTU (benzotriazole-N, N, N ', N '-tetramethyl isourea hexafluorophosphate): NMM (N-
Methylmorpholine)=1: 3: 2.85: 6.It is sequentially connected 1- naphthylalanine, 4- amine methyl cyclohexane formic acid, benzene according to the method described above
The amino acid of 3 equivalents is added in alanine, and appropriate DMF is added, and reacts 1h.Synthetic product is transferred in cutting pipe, E liquid is added,
Shake cleavage reaction 1h；Cutting liquid is collected into centrifuge tube, 6 times of volume ice ether, low speed centrifuge precipitating is added；It will precipitating
Crude product washed three times with ether, obtain the crude product of precursor compound 5.Compound is obtained by partly preparing chromatography
5PSMA-Ala(Nap)-Cyh-Phe。
The preparation of compound 6
The synthesis of PSMA-Ala (Nap)-Cyh-Phe-NOTA (compound 6) uses solid-phase synthesis.According to compound 5
Preparation method sequentially adds 3 equivalent chelating agent NOTA on the basis of compound 5, and appropriate DMF is added, and reacts 1h.It will synthesis
Product is transferred in cutting pipe, and E liquid is added, and shakes cleavage reaction 1h；Cutting liquid is collected into centrifuge tube, 6 times of volumes are added
Ice ether, low speed centrifuge precipitating；The crude product of precipitating is washed three times with ether.Compound is obtained by partly preparing chromatography
6。
(S2) it is carried out using direct labelling method68Ga radioactive label；
It opens reactor and temperature is set in 95 DEG C；The NOTA-ANCP-PSMA lyophilized preparation of freezing is taken out, use is ultrapure
Water is configured to 5mg/mL solution, and 4 μ L is taken to be placed in 1.5mL EP reaction tube, and 90 μ L 1.0mol/LHEPES are added into reaction tube
Buffer solution.3.5mL 0.05mol/L HCl solution is taken, is eluted68Ge-68Ga generator (preceding 1.5mL leacheate can discard,
Take rear 2.0mL use)；1.0mL is taken to elute68Reaction tube, juxtaposition is added in Ga 0.05mol/L HCl solution, about 111-185MBq
In 95 DEG C of heating module.15min is reacted under 200rpm revolving speed.Solution after reaction is taken out with 2mL syringe and uses work
Degree meter measurement total activity, accurate pH test paper measure pH value, are then injected into the Sep-pak C18 column of activation, with 3.0mL pure water
C18 column is rinsed, to clean impurity；0.22 μm of miillpore filter is installed on a cl 8 column, is eluted with the ethanol solution of 1.0mL 95%,
It is collected into Sterile vacuum bottle.When for zoopery, the group of 95% ethanol rinse is placed on heating module, with nitrogen
Drying, is redissolved with physiological saline, obtains target label object68Ga-NOTA-ANCP-PSMA。
Using its structure of mass spectrum confirmation, mark rate and radiochemical purity are measured respectively using HPLC.
Mass spectral results as shown in Figure 1, in negative ion mass spectrum figure as can be seen that 542.8 at be the peak (M-2H)/2,1086.3
Place is the peak M-H, and molecular weight actually 1087.5, according to mass spectrum confirmation, its structure is correct.
As shown in Fig. 2, the retention time of precursor compound NOTA-ANCP-PSMA is 11.783min or so.It can from Fig. 3
To find out, retention time is 12.138min or so to the compound after label in radioactive detector, the guarantor with precursor compound
Stay time consistency.Its radiochemical purity also up to 95.2%, is dissociated68Ga3+Peak area ratio is 1.0%, and impurity is polypeptide impurities
The miscellaneous peak of formation, radiochemical purity reach 95% or more.
Synthetic route is as follows:
Embodiment 2
Vitro stability measurement:
68The radiochemical stability of Ga-NOTA-ANCP-PSMA is opened in two kinds of systems of calf serum and phosphate buffer
Exhibition.PBS method: being placed in the phosphate buffer (PBS, pH=7.4) of 0.5mL, be placed at 37 DEG C, be incubated for 30,60,90,120,
After 150min, its radiochemical purity is measured using HPLC, to measure its vitro stability.Serum method: it is placed in the calf of 0.5mL
In serum solution, it is incubated under the conditions of pH=7,37 DEG C.When being incubated for 30,60,90,120,150min, it is measured using HPLC and is put
Chemical purity is penetrated, to measure its vitro stability.
As shown in figure 4,68Ga-NOTA-ANCP-PSMA having good stability in PBS system, until 150min radiochemistry
Purity maintains 95.4% or more always.Vitro stability in BSA system the result shows that, the radiochemically pure in 90min
Degree can maintain 95% or more level, be declined slightly later.Explanation68Stability of the Ga-NOTA-ANCP-PSMA in BSA
Relative to requirement poor in PBS, but can satisfy PET imaging.
Embodiment 3
The measurement of lipid:
Marker68Ga-NOTA-ANCP-PSMA isolates and purifies to obtain by Sep-Pak C18Cartridge, uses 95%
Ethanol elution.The marker of acquisition is dried with nitrogen by dry, using same volume (0.5mL:0.5mL) n-octyl alcohol and
Obtained marker is dissolved in 1.5mL EP pipe (about 3.7MBq) by phosphate buffer solution (pH=7.4).5min fullys shake,
Centrifugation layering 5min, revolving speed 2000rpm in centrifuge.Take organic phase and each 100uL of water phase in 1mL EP pipe respectively, in well
Its radiocounting is measured in type gamma detector respectively, rouge moisture is calculated by the ratio of the radiocounting of organic phase and water phase
Distribution coefficient P.P=log (No/NW)(NoAnd NWThe respectively counting of organic phase and aqueous sample).It repetitive operation 3 times, is averaged
For the lipid of the marker.
It is calculated according to measured data, lipid is -1.36 ± 0.02.
Embodiment 4
The bio distribution of tumor bearing nude mice:
LNCaP tumor mass is inoculated under 7-8 weeks nude mice right axillary, in 1 to 2 week of tumour growth, reaches major diameter 0.8-
1.0cm using.Tumor bearing nude mice 12 are taken, 3 one group, takes 0.15mL's (111-185kBq)68Ga-NOTA-ANCP-PSMA passes through
Tail vein injection.In 5min 15min, 30min, the 60min neck that breaks are put to death, and distinguish that coring is dirty, lung, liver,spleen,kidney, pancreas after execution
The organs such as gland, stomach, small intestine, large intestine, bladder, bone, muscle, tumour, and carry out taking blood.Each organ is weighed, activity meter
Number counts and carries out standard correction by titer, by ID% is calculated.
The results are shown in Table 1,68Ga-NOTA-ANCP-PSMA is removed comparatively fast in blood, mainly passes through renal metabolism, liver
It is dirty intake it is lower, absorbed in heart, lung, spleen, pancreas, stomach, small intestine, large intestine it is lower, far below blood absorb；Muscle, bone
It is relatively low Deng absorbing.Prolonged high intake is not shown in blood, blood is removed comparatively fast, shown68The compound of Ga label
Stability is preferable in vivo, does not generate free Ga3+It is enriched in blood.There is higher intake in tumour, in 5-15min
Left and right intake reaches peak, gradually decreases later.
Table 168The tumor bearing nude mice bio distribution result of Ga-NOTA-ANCP-PSMA
Embodiment 5
PET-CT imaging:
When MicroPET imaging research, take 0.15mL's (about 3.7MBq)68Ga-NOTA-ANCP-PSMA is infused by tail vein
It is mapped in the mouse of lotus LNCaP tumor.The postanesthetic animal prone type of isoflurane is fixed in animal PET scanner, after injection
Start within 60 minutes to scan 15 minutes static state PET-CT images.
After injection 60min PET-CT as the result is shown (Fig. 5), due to68Ga-NOTA-ANCP-PSMA is mainly drained by urine,
Still there is higher intake in kidney and bladder, have obvious intake in tumor locus, shows higher target/non-target ratio.Due to blood medicine
Concentration decline, the non-target organ intake such as heart, lung, liver are decreased obviously, and background tissues and organ do not have substantially in addition to kidney, bladder
There is radioactive uptake, preferable image can be provided in the high intake of 60min.
Claims (7)
1. a kind of prostate cancer PET diagnostic reagent68Ga-NOTA-ANCP-PSMA, which is characterized in that described68Ga-NOTA-ANCP-
The concrete structure formula of PSMA is as follows:
2. a kind of compound N OTA-ANCP-PSMA, which is characterized in that the concrete structure formula of the NOTA-ANCP-PSMA is as follows:
3. a kind of prostate cancer PET diagnostic reagent as described in claim 168The preparation method of Ga-NOTA-ANCP-PSMA,
It is characterized in that, described method includes following steps:
(S1) precursor compound NOTA-ANCP-PSMA is synthesized using solid-phase synthesis；
The preparation of compound 3:
Weighing degree of substitution is 0.3mmol/g starting material Fmoc-Lys (Dde)-Wang Resin3 g, is added into reactor,
DMF is added, impregnates 30min；DMF is drained later, adds the 20%Pip/DMF of 3 times of volumes, nitrogen charging gas shielded, to remove Fmoc,
30min is reacted, 20%Pip/DMF is drained, DMF is washed 5 times, ninhydrin detection display navy blue；It is put in proportion into N, N'- amber
Imide carbonic ester DSC, n,N-diisopropylethylamine DIPEA and 4-dimethylaminopyridine DMAP, are added appropriate DMF, in nitrogen
Under gas shielded, 1h is reacted；H-Glu (OtBu)-OtBuHCl is added into reactor, appropriate DMF is added, in nitrogen protection
Under, 1h is reacted, compound 3 is obtained；
The preparation of compound 4:
By the cutting liquid E liquid cleavage reaction 1h of compound 3, determine whether to be fully connected using ninhydrin detection method, if not completely
Connection is fed intake again until being fully connected；After being fully connected, 3 times of 2% hydrazine hydrates of volume, DMF solutions are added, react 30min,
Dde protection is sloughed, DMF is washed 5 times, obtains compound 4；
The preparation of compound 5:
Amino acid, compound 4, benzotriazole-N, N, N needed for side chain is added according to polypeptide solid-state reaction method ', N '-tetramethyl
Base isourea hexafluorophosphate HBTU, N- methylmorpholine NMM；It is sequentially connected 1- naphthylalanine, 4- amine first according to the method described above
The amino acid of 3 equivalents is added in base cyclohexanecarboxylic acid, phenylalanine, and appropriate DMF is added, and reacts 1h；Synthetic product is transferred to cutting
E liquid is added in Guan Zhong, shakes cleavage reaction 1h；Cutting liquid is collected into centrifuge tube, 6 times of volume ice ether, low-speed centrifugal is added
Machine precipitating；The crude product of precipitating is washed three times with ether, obtains the crude product of precursor compound 5；By partly preparing chromatography
Obtain compound 5；
The preparation of compound 6:
On the basis of compound 5, the chelating agent NOTA of 3 equivalents is added, appropriate DMF is added, reacts 1h；Synthetic product is shifted
Into cutting pipe, E liquid is added, shakes cleavage reaction 1h；Cutting liquid is collected into centrifuge tube, 6 times of volume ice ether are added, it is low
Fast centrifuge precipitating；The crude product of precipitating is washed three times with ether；Compound 6 is obtained by partly preparing chromatography；
The compound 6 is precursor compound NOTA-ANCP-PSMA；
(S2) it is carried out using direct labelling method68Ga radioactive label；
It opens reactor and temperature is set in 95 DEG C；The 6 NOTA-ANCP-PSMA lyophilized preparation of compound of freezing is taken out, is used
Ultrapure water is configured to 5mg/mL solution, and 4 μ L is taken to be placed in 1.5mL EP reaction tube, and 90 μ L1.0mol/L are added into reaction tube
HEPES buffer solution；3.5mL0.05mol/L HCl solution is taken, is eluted68Ge-68Ga generator；1.0mL is taken to elute68Ga0.05mol/L HCl solution, about 111-185 MBq are added reaction tube, are placed in 95 DEG C of heating module；200rpm turns
Speed is lower to react 15min；Solution after reaction is taken out and measures total activity with activity meter, pH value is measured, is then injected into activation
Sep-Pak C18 column in, with 3.0mL pure water rinsing C18 column, to clean impurity；0.22 μm of micropore filter is installed on a cl 8 column
Film is eluted with the ethanol solution of 1.0mL95%, is collected into Sterile vacuum bottle；When for zoopery, 95% ethyl alcohol is drenched
The group washed is placed on heating module, to be dried with nitrogen, is redissolved with physiological saline, obtains target label object68Ga-NOTA-ANCP-
PSMA；
Synthetic route are as follows:
4. prostate cancer PET diagnostic reagent according to claim 368The preparation method of Ga-NOTA-ANCP-PSMA, it is special
Sign is, in the preparation of the compound 3, the molar ratio of Fmoc-Lys (Dde)-Wang Resin, DSC, DIPEA, DMAP
For 1:6:12:1；
Fmoc-Lys (Dde)-Wang Resin, H-Glu (OtBu)-OtBuHCl, DIPEA, DMAP molar ratio be 1:3:9:
1。
5. prostate cancer PET diagnostic reagent according to claim 368The preparation method of Ga-NOTA-ANCP-PSMA, it is special
Sign is, in the preparation of compound 4, pure TFA in the cutting liquid E liquid, thioanisole, water, phenol, 1,2- dithioglycol
Mol ratio is 35:2:1:1:1.
6. prostate cancer PET diagnostic reagent according to claim 368The preparation method of Ga-NOTA-ANCP-PSMA, it is special
Sign is, in the preparation of the compound 5, compound 4, amino acid, benzotriazole-N, N, N ' and, N '-tetramethyl isourea six
Fluorophosphate, N- methylmorpholine molar ratio be 1:3:2.85:6.
7. a kind of prostate cancer PET diagnostic reagent as described in claim 168Ga-NOTA-ANCP-PSMA is examined in prostate cancer
It is disconnected, by stages with the application in therapeutic evaluation.
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