CN107501393A - 18F labeled amino acid polypeptide drug synthetic methods and kit - Google Patents

18F labeled amino acid polypeptide drug synthetic methods and kit Download PDF

Info

Publication number
CN107501393A
CN107501393A CN201710821964.6A CN201710821964A CN107501393A CN 107501393 A CN107501393 A CN 107501393A CN 201710821964 A CN201710821964 A CN 201710821964A CN 107501393 A CN107501393 A CN 107501393A
Authority
CN
China
Prior art keywords
nota
bottle
alf
pillar
amino acid
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201710821964.6A
Other languages
Chinese (zh)
Other versions
CN107501393B (en
Inventor
唐刚华
胡晓平
胡生焰
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
BEIJING PET Co Ltd
Original Assignee
BEIJING PET Co Ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by BEIJING PET Co Ltd filed Critical BEIJING PET Co Ltd
Priority to CN201710821964.6A priority Critical patent/CN107501393B/en
Publication of CN107501393A publication Critical patent/CN107501393A/en
Application granted granted Critical
Publication of CN107501393B publication Critical patent/CN107501393B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D255/00—Heterocyclic compounds containing rings having three nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D249/00 - C07D253/00
    • C07D255/02—Heterocyclic compounds containing rings having three nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D249/00 - C07D253/00 not condensed with other rings

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

The present embodiments relate to one kind18F labeled amino acid polypeptide drug synthetic methods and kit, methods described include：The capture cyclotron production of anion pillar18F‑Ion；0.4 0.5ml physiological saline after anion pillar by eluting18F‑Ion obtains the first reaction liquid, into reaction bulb；By precursor AlCl of the pH value between 453Acid solution organic solvent mixing liquid adds reaction bulb；Precursor is NOTA derivatives；NOTA is the alkyl triacetic acid base of three azacyclo- nine or the alkyl diacetoxyl of three azacyclo- nine；Synthesis is marked to 100 DEG C 110 DEG C in heating response bottle；After cooling, liquid in SPE reaction bulb is diluted with water；Liquid after being diluted in SPE reaction bulb is separated by solid phase extraction column so that product absorption to be obtained is on solid phase extraction column；Solid phase extraction column is cleaned with water, removes organic solvent and impurity；With ethanol rinse solid phase extraction column, product to be obtained is washed into product bottle, and add preparation liquid to obtain synthesized product.

Description

18F labeled amino acid polypeptide drug synthetic methods and kit
Technical field
The present invention relates to pharmaceutical synthesis field, more particularly to one kind18F labeled amino acid polypeptide drug synthetic methods and examination Agent box.
Background technology
18F- fluorodeoxyglucoses (18F-FDG early diagnosis of tumor and guiding treatment) are widely used to, but it is one (, easily there is false positive and false negative result to the non-specific positron emission fault of kind in PET medicines.18F labeled amino acids and targeting Receptor-specific18F labeling polypeptide class PET medicines can make up18F-FDG wretched insufficiency, it is the important development for developing PET medicines Direction.It is however, existing general18F mark biomolecule methods are related to N- succinimides -4-18F- fluorobenzoates (18F- SFB) and18F-4- nitrobenzophenone -2- fluoropropionic acids ester (18F-NFP) labeled amino acid polypeptide-NH2 bases, reactions steps are more, operation It is more complicated, generated time length, it is not easy to realize Fully automated synthesis.Simple and practical Al18F labelling methods turn into18F labeled amino acid polypeptides The inevitable choice of class medicine amino.
With mark prothetic group18F-SFB and18It is prepared by F-NFP18F- mark bioactive molecule amino is conventional method the most, but The method is complicated, putting low yield, it is difficult to completes industrialization and produces automatically.B-18F or Si-18Although F one-step method can high-efficient automatic Prepare18F- marks bioactive molecule, but what the method obtained is to contain carrier18F- labeled amino acid polypeptide drugs.
18F-AlF methods have been used for preparing targeting somatostatin receptor18Alkyl diacetoxyl -1- the third ammonia of naphthalene of tri- azacyclo-s of F- nine Acid-Octreotide (18) and targeted integration element α v β 3 F-AlF-NOTA-NOC18The PVOH of tri- azacyclo-s of F-, nine alkyl triacetic acid base-three Amido-cyclic divalent arginine-glycine-aspartic acid (18F-AlF-NOTA-PEG3-Glu-RGD2),18F-AlF-NOTA- NOC and18F-AlF-NOTA-PEG3-Glu-RGD2 shows preferable potential applicability in clinical practice.Recently, we also have developed targeted integration Plain α v β 318F marks betas-glutamic acid coupling divalence symmetry18F-AlF-NOTA-PEG3- β-Glu-RGD2 and18The azacyclo-s of F- tri- Nine alkyl diacetoxyls-Pidolidone (18F-AlF-NOTA-Glu), and it is applied to a variety of lotus knurl model mouse PET imaging and grinds Study carefully, show preferable application potential.It is but existing18F-AlF labeled amino acid polypeptide drugs, such as18F-AlF-NOTA-Glu 、18F-AlF-NOTA-NOC、18F-AlF-NOTA-PEG3-Glu-RGD2 and18F-AlF-NOTA-PEG3- β-Glu-RGD2 etc., only Low dosage can be prepared using RNA isolation kit18F-AlF-NOTA- biomolecule, it is impossible to will be accelerator-produced all18F-Ion is straight Connect and be used for18F-AlF labeled amino acids and polypeptide, it is difficult to realize full-automatic industrialization production, and one is brought to radiological safety protection Determine risk.
The content of the invention
The purpose of the present invention is the defects of being directed to prior art, there is provided Yi Zhongyong18F labeled amino acids polypeptide drug synthesizes Method and kit.
In view of this, in a first aspect, the embodiments of the invention provide one kind18F labeled amino acid polypeptide drugs synthesis side Method, including：
The capture cyclotron production of anion pillar18F-Ion；
0.4-0.5ml physiological saline or other inorganic salt solutions after anion pillar by eluting18F-Ion obtains First reaction liquid, into reaction bulb；
By precursor-AlCl of the pH value between 4-53Acid solution-organic solvent mixing liquid adds reaction bulb；Before described Body is NOTA derivatives；The NOTA is specially the alkyl triacetic acid base of three azacyclo- nine or the alkyl diacetoxyl of three azacyclo- nine；
Synthesis is marked to 100 DEG C -110 DEG C in heating response bottle；
After cooling, liquid in the reaction bulb is diluted with water；
Liquid after being diluted in the reaction bulb is separated by solid phase extraction column so that product absorption to be obtained On solid phase extraction column；
Solid phase extraction column is cleaned with water, removes organic solvent and impurity；
With ethanol rinse solid phase extraction column, product to be obtained is washed into product bottle, and add prepare liquid obtain it is synthesized Product.
Preferably, the anion pillar is specially SEP-PAK Light QMA pillars.
Preferably, the precursor-AlCl3Than for 1.2:1.0, the quality of the precursor is 200-300 μ g.
Preferably, the solid phase extraction column is specially C18 pillars or HLB pillars.
Preferably, the precursor is the alkyl diacetoxyl of three azacyclo- nine-Pidolidone NOTA-Glu, and described first reacts Liquid is18F-AlF, the product are18The alkyl diacetoxyl of tri- azacyclo-s of F- nine-Pidolidone18F-AlF-NOTA-Glu。
Preferably, the precursor is the alkyl diacetoxyl -1- naphthylalanines of three azacyclo- nine-Octreotide NOTA-NOC, institute Stating the first reaction liquid is18F-AlF, the product are18Alkyl diacetoxyl -1- the naphthylalanines of tri- azacyclo-s of F- nine-Austria is bent Peptide18F-AlF-NOTA-NOC。
Preferably, the precursor is the Polyethylenimine base-β of three azacyclo-s, nine alkyl triacetic acid base-three-glutamic acid base-ring-type Divalence arginine-glycine-aspartic acid NOTA-PEG3- β-Glu-RGD2, first reaction liquid are18F-AlF, it is described Product is18The Polyethylenimine base of tri- azacyclo-s of F-, nine alkyl triacetic acid base-three-cyclic divalent arginine-glycine-aspartic acid18F-AlF-NOTA-PEG3-β-Glu-RGD2。
Preferably, it is described18F labeled amino acid polypeptide drug synthetic methods are used for18F-AlF-NOTA- biomolecule Full-automatic industrialization production.
Second aspect, the embodiments of the invention provide one kind to be used for18The examination of F labeled amino acid polypeptide drug synthetic methods Agent box, the kit include：The first reagent bottle equipped with 0.4ml-0.5ml physiological saline, equipped with 0.4ml-1.2ml precursors- AlCl3 the second reagent bottle and pH is the sub- reagent bottles of 4-5 acidity acetonitrile solutions, the 3rd reagent bottle equipped with 15ml water, is equipped with 4th reagent bottle of 10ml water, the 5th reagent bottle equipped with 0.5-1.5ml ethanol, equipped with 10-18ml ascorbic acid containing 0.57mM Physiological saline the 6th reagent bottle, anion pillar, solid phase extraction column, anion pillar, prepared microcolumn solvent or molten Liquid, syringe, syringe needle, receiving bottle and inlet and outlet sterilised membrane filter.
Preferably, the anion pillar is specially SEP-PAK Light QMA pillars；The solid phase extraction column is specific For C18 pillars or HLB pillars；The prepared microcolumn solvent or solution specifically include ethanol, water for injection and sodium acid carbonate Solution.
It is provided in an embodiment of the present invention18F labeled amino acid polypeptide drug synthetic methods, cyclotron production18F- After being trapped by anion QMA pillars, eluted with a small amount of physiological saline18F-, the whole of elution18F-Using18It is prepared by F-AlF methods18F- AlF-NOTA- bioactive molecules, the method be referred to as QMA trapping-18F-AlF methods.QMA trappings-18F-AlF methods can not only be completed entirely Automated radio synthesizes, and can realize quick, efficient, high dose industrialization production using kit18F-AlF-NOTA- biologies Bioactive molecule, it is18The GMP productions of F- mark bioactive molecules and clinical Transformation Application are laid a good foundation, and can fully meet city Field and growing number of patient P ET, which are checked, to be needed.QMA trappings-18The Transformation Application of F-AlF methods will open up18F- mark biologies are living Property molecule research and development and the clinical Transformation Application new situation, promote the development of accurate diagnosis and treatment, and huge social benefit and society will be produced Can benefit.
Brief description of the drawings
Fig. 1 is provided in an embodiment of the present invention18The process flow diagram of F labeled amino acids polypeptide drug synthesis；
Fig. 2 a are provided in an embodiment of the present invention18F-AlF-NOTA-Glu building-up process schematic diagram；
Fig. 2 b are provided in an embodiment of the present invention18F-AlF-NOTA-NOC building-up process schematic diagram；
Fig. 2 c are provided in an embodiment of the present invention18F-AlF-NOTA-PEG3- β-Glu-RGD2 building-up process schematic diagram；
Fig. 3 a are provided in an embodiment of the present invention18F-AlF-NOTA-NOC high performance liquid chromatography (HPLC) analysis collection of illustrative plates；
Fig. 3 b are provided in an embodiment of the present invention18F-AlF-NOTA-NOC thin-layer chromatography (TLC) analysis collection of illustrative plates；
Fig. 3 c are provided in an embodiment of the present invention18F-AlF-NOTA-PEG3- β-Glu-RGD2 HPLC analysis collection of illustrative plates；
Fig. 3 d are provided in an embodiment of the present invention18F-AlF-NOTA-PEG3- β-Glu-RGD2 TLC analysis collection of illustrative plates；
Fig. 3 e are provided in an embodiment of the present invention18F-AlF-NOTA-Glu HPLC analysis collection of illustrative plates；
Fig. 3 f are provided in an embodiment of the present invention18F-AlF-NOTA-Glu TLC analysis collection of illustrative plates.
Fig. 4 A are the double knurl models of lotus provided in an embodiment of the present invention18F-AlF-NOTA-PEG3- β-Glu-RGD2 positive electrons are sent out Penetrate fault imaging PET image
Fig. 4 B are intact animal provided in an embodiment of the present invention18F-AlF-NOTA-NOC PET image；
Fig. 4 C are intact animal provided in an embodiment of the present invention18F-AlF-NOTA-Glu PET image.
Embodiment
Below by drawings and examples, technical scheme is described in further detail.
18F labeled amino acids polypeptide drug synthetic method provided in an embodiment of the present invention, its process flow diagram is such as Shown in Fig. 1.Produced by cyclotron18F-Ion, captured by anion pillar (QMA is identified as in figure)18F-Ion；Then By 0.4-0.5ml physiological saline by being eluted after anion pillar18F-Ion obtains the first reaction liquid, into reaction bulb 10；Also, by precursor-AlCl of the pH value between 4-53Acid solution-organic solvent mixing liquid adds reaction bulb 10；Its In, precursor is NOTA derivatives；NOTA is the alkyl triacetic acid base of three azacyclo- nine or the alkyl diacetoxyl of three azacyclo- nine；Then 10 to 100 DEG C -110 DEG C of heating response bottle, chelation reaction occurs, synthesis is marked；After cooling, liquid in reaction bulb 10 is entered Row is diluted with water；Liquid after being diluted in reaction bulb 10 is separated by solid phase extraction column (C-18 is identified as in figure), So that product absorption to be obtained is on solid phase extraction column；Solid phase extraction column is cleaned with water, removes organic solvent and impurity；With Ethanol rinse solid phase extraction column, product to be obtained is washed into product bottle 9, and add preparation liquid to obtain synthesized product.
Wherein, anion pillar can specifically use SEP-PAK Light QMA pillars；Precursor-AlCl3Than for 1.2: 1.0, the quality of precursor is 200-300 μ g；Solid phase extraction column can specifically use C18 pillars or HLB pillars.
In a specific example, precursor is the alkyl diacetoxyl of three azacyclo- nine-Pidolidone (NOTA-Glu), the One reaction liquid is18F-AlF, product are18The alkyl diacetoxyl of tri- azacyclo-s of F- nine-Pidolidone (18F-AlF-NOTA- Glu).The building-up process schematic diagram is as shown in Figure 2 a.
In another specific example, precursor is the alkyl diacetoxyl -1- naphthylalanines of three azacyclo- nine-Octreotide (NOTA-NOC), the first reaction liquid is18F-AlF, product are18Alkyl diacetoxyl -1- the naphthylalanines of tri- azacyclo-s of F- nine - Octreotide (18F-AlF-NOTA-NOC).The building-up process schematic diagram is as shown in Figure 2 b.
In another specific example, precursor is the Polyethylenimine base-β of three azacyclo-s, nine alkyl triacetic acid base-three-paddy ammonia Acidic group-cyclic divalent arginine-glycine-aspartic acid NOTA-PEG3- β-Glu-RGD2, the first reaction liquid are18F-AlF, Product is18The Polyethylenimine base of tri- azacyclo-s of F-, nine alkyl triacetic acid base-three-cyclic divalent arginine-glycine-aspartic acid (18F-AlF-NOTA-PEG3-β-Glu-RGD2).The building-up process schematic diagram is as shown in Figure 2 c.
The present embodiment18F labeled amino acid polypeptide drug synthetic methods can be used for18F-AlF-NOTA- biomolecule Full-automatic industrialization production.
For ease of the implementation of this method, the invention also provides a kind of kit, including：Equipped with 0.4ml-0.5ml physiology First reagent bottle of salt solution, the second reagent bottle equipped with 0.4ml-1.2ml precursors-AlCl3 and pH are 4-5 acidity acetonitrile solutions Sub- reagent bottle, the 3rd reagent bottle equipped with 15ml water, the 4th reagent bottle equipped with 10ml water, equipped with 0.5-1.5ml ethanol Five reagent bottles, the 6th reagent bottle of physiological saline equipped with 10-18ml ascorbic acid containing 0.57mM, anion pillar, solid phase extraction Take pillar, anion pillar, prepared microcolumn solvent or solution, syringe, syringe needle, receiving bottle and inlet and outlet sterilised membrane filter.
Likewise, in kit, anion pillar is specially SEP-PAK Light QMA pillars；Solid phase extraction column Specially C18 pillars or HLB pillars；Prepared microcolumn solvent or solution specifically include ethanol, water for injection and sodium acid carbonate Solution.
Below, with some specific embodiments, it is further elaborated.
Embodiment 118The full-automatic industry of F-AlF-NOTA-Glu parenteral solutions is combined to
Kit forms：0.4-0.5mL physiological saline (bottle 1), the μ L of 200-300 μ g precursors NOTA-Glu+2mM 24 AlCl3(bottle 2), 0.8-1.2mL pH4-5 acidity acetonitrile solutions (bottle 2 ') (bottle 2 ' is not shown in Fig. 1 in kit, The solution of bottle 2 ' is added in bottle 2 before the synthesis), 15mL waters for injection (bottle 3), 10mL waters for injection (bottle 4), 1.5mL ethanol The physiological saline (bottle 6) of (bottle 5) and 18mL ascorbic acid containing 0.57mM.Auxiliary material includes：SEP-PAK C18 separation pillar, SEP-PAK QMA pillars, pillar pre-treatment solvents or solution (ethanol, water for injection, 8.4% sodium bicarbonate solution), syringe With syringe needle, receiving bottle, inlet and outlet sterilised membrane filter.SEP-PAK QMA pillars, or other anion pillars replace；Physiology salt Water can be replaced with other inorganic salt solutions；Acetonitrile can be replaced with dimethyl sulfoxide (DMSO), ethanol and methanol；SEP-PAK C18 are small Post, it is also possible to which HLB pillars replace.
Design18F-AlF marks Fully automated synthesis program, and it is small-sized to start self-control18Before F-FDG synthesizers (as shown in Figure 1), Acid acetonitrile solution in bottle 2 ' is added in bottle 2；SEP-PAK QMA pillars are successively with 8.4% sodium bicarbonate solution and injection Water pretreatment；SEP-PAK C18 separation pillars are pre-processed with ethanol and water for injection successively；By bottle 1, bottle 2, bottle 3, bottle 4, bottle 5, Bottle 6, SEP-PAK QMA pillars, SEP-PAK C18 separation pillars and the receiving bottle with inlet and outlet sterilised membrane filter are connected to conjunction Cheng Yi (as shown in Figure 1) corresponding position.
Start synthesizer, start to produce18F-AlF-NOTA-Glu parenteral solutions.Passed through by cyclotron18O(p,n)18F cores Reaction production18F-, in N2Under airborne band, by the Sep-Pak QMA anion pillars being placed in radioactive activity measuring meter,18F- It is trapped in pillar,18O- water is collected in returnable bottle.In N2Under carrier band, physiological saline is by QMA pillars in bottle 118F-Wash Take off in confined reaction bottle 10.Then, in N2Under carrier band, the μ L AlCl of NOTA-Glu+2mM 24 in bottle 23Solution is added into instead Answer in bottle 10, mix, 100-110 DEG C of heating response 10min.After cooling, 15mL waters for injection are added into reaction bulb 10 in bottle 3 In, after mixing, it is transferred to SEP-PAK C18 pillars.After solution has all shifted in reaction bulb, 10mL waters for injection rush in bottle 4 Pillar is washed, and uses N2Dry up pillar.Finally, product is eluted in receiving bottle 8 by ethanol in bottle 5, and normal saline flushing is small in bottle 6 Post cut-back product, obtain satisfactory18F-AlF-NOTA-Glu parenteral solutions.18F-AlF-NOTA-Glu does not correct radiochemistry Yield is 40%-60%.
Embodiment 218The full-automatic industry of F-AlF-NOTA-NOC parenteral solutions is combined to
Kit forms：0.4-0.5mL physiological saline (bottle 1), the μ L of 200-300 μ g precursors NOTA-NOC+2mM 24 AlCl3(bottle 2), 0.8-1.2mLpH4-5 acidity acetonitrile solution (bottle 2 '), 15mL physiological saline (bottle 3), 10mL physiological saline (bottles 4), the physiological saline (bottle 6) of 1.5mL ethanol (bottle 5) and 18mL ascorbic acid containing 0.57mM.Auxiliary material includes：HLB pillars, (ethanol, physiological saline, water for injection, 8.4% sodium acid carbonate are molten for SEP-PAK QMA pillars, pillar pre-treatment solvents or solution Liquid), syringe and syringe needle, receiving bottle, inlet and outlet sterilised membrane filter.SEP-PAK QMA pillars, or other anion pillar generations Replace；Physiological saline can be replaced with other inorganic salt solutions；Acetonitrile can be replaced with dimethyl sulfoxide (DMSO), ethanol and methanol；HLB is small Post, it is also possible to which SEP-PAK C18 separation pillars replace.
Design18F-AlF marks Fully automated synthesis program, and it is small-sized to start self-control18Before F-FDG synthesizers (as shown in Figure 1), Acid acetonitrile solution in bottle 2 ' is added in bottle 2；SEP-PAK QMA pillars are successively with 8.4% sodium bicarbonate solution and injection Water pretreatment；HLB pillars are pre-processed with ethanol and physiological saline successively；By bottle 1, bottle 2, bottle 3, bottle 4, bottle 5, bottle 6, SEP-PAK QMA pillars, HLB pillars and the receiving bottle with inlet and outlet sterilised membrane filter are connected to synthesizer (Fig. 1) corresponding position.
Start synthesizer, start to produce18F-AlF-NOTA-NOC parenteral solutions.Passed through by cyclotron18O(p,n)18F cores Reaction production18F-, in N2Under airborne band, by the Sep-Pak QMA anion pillars being placed in radioactive activity measuring meter,18F- It is trapped in pillar,18O- water is collected in returnable bottle.In N2Under carrier band, physiological saline is by QMA pillars in bottle 118F-Wash Take off in confined reaction bottle 10.Then, in N2Under carrier band, the μ L AlCl of NOTA-NOC+2mM 24 in bottle 23Solution is added into instead Answer in bottle 10, mix, 100-110 DEG C of heating response 10min.After cooling, 15mL physiological saline is added into reaction bulb 10 in bottle 3 In, after mixing, it is transferred in HLB pillars.After solution has all shifted in reaction bulb, 10mL normal saline flushings pillar in bottle 4, And use N2Dry up pillar.Finally, product is eluted in receiving bottle 8 by ethanol in bottle 5, and normal saline flushing pillar dilutes in bottle 6 Product, obtain satisfactory18F-AlF-NOTA-NOC parenteral solutions.18F-AlF-NOTA-NOC does not correct Radiochemical yield 40%-60%.
Embodiment 318The full-automatic industry of F-AlF-NOTA-PEG3- β-Glu-RGD2 parenteral solutions is combined to
Kit forms：0.4-0.5mL physiological saline (bottle 1), 200-300 μ g precursor NOTA-PEG3- β-Glu-RGD2+ 2mM 24μL AlCl3(bottle 2), 0.8-1.2mLpH4-5 acidity acetonitrile solution (bottle 2 '), 15mL waters for injection (bottle 3), 10mL notes Penetrate the physiological saline (bottle 6) with water (bottle 4), 1.5mL ethanol (bottle 5) and 18mL ascorbic acid containing 0.57mM.Auxiliary material includes： SEP-PAK C18 separation pillar, SEP-PAK QMA pillars, pillar pre-treatment solvents or solution (ethanol, water for injection, 8.4% Sodium bicarbonate solution), syringe and syringe needle, receiving bottle, inlet and outlet sterilised membrane filter.SEP-PAK QMA pillars, or other the moon Ion pillar replaces；Physiological saline can be replaced with other inorganic salt solutions；Acetonitrile can use dimethyl sulfoxide (DMSO), ethanol and methanol Instead of；SEP-PAK C18 pillars, it is also possible to which HLB pillars replace.
Design18F-AlF marks Fully automated synthesis program, and it is small-sized to start self-control18Before F-FDG synthesizers (as shown in Figure 1), Acid acetonitrile solution in bottle 2 ' is added in bottle 2；SEP-PAK QMA pillars are successively with 8.4% sodium bicarbonate solution and injection Water pretreatment；SEP-PAK C18 separation pillars are pre-processed with ethanol and water for injection successively；By bottle 1, bottle 2, bottle 3, bottle 4, bottle 5, Bottle 6, SEP-PAK QMA pillars, SEP-PAK C18 separation pillars and the receiving bottle with inlet and outlet sterilised membrane filter are connected to conjunction Cheng Yi (as shown in Figure 1) corresponding position.
Start synthesizer, start to produce18F-AlF-NOTA-PEG3- β-Glu-RGD2 parenteral solutions.Led to by cyclotron Cross18O(p,n)18F nuclear reactions production18F-, in N2It is cloudy by the Sep-Pak QMA being placed in radioactive activity measuring meter under airborne band Ion pillar,18F-It is trapped in pillar,18O- water is collected in returnable bottle.In N2Under carrier band, physiological saline will in bottle 1 In QMA pillars18F-Elute in confined reaction bottle 10.Then, in N2Under carrier band, NOTA-PEG3- β-Glu-RGD2+ in bottle 2 2mM 24μL AlCl3Solution is added into reaction bulb 10, is mixed, 100-110 DEG C of heating response 10min.After cooling, in bottle 3 15mL waters for injection are added into reaction bulb 10, after mixing, are transferred to SEP-PAK C18 pillars.Solution is whole in reaction bulb After having shifted, 10mL waters for injection rinse pillar in bottle 4, and use N2Dry up pillar.Finally, product is eluted to by ethanol in bottle 5 In receiving bottle 8, normal saline flushing pillar cut-back product in bottle 6, obtain satisfactory18F-AlF-NOTA-PEG3-β-Glu- RGD2 parenteral solutions.18It is 20%-40% that F-AlF-NOTA-PEG3- β-Glu-RGD2, which do not correct Radiochemical yield,.
The measure of the product purity of embodiment 4
Determined with radioactivity high performance liquid chromatography (HPLC) and thin-layered chromatography (TLC)18F-AlF-NOTA-Glu、18F- AlF-NOTA-NOC and18F-AlF-NOTA-PEG3- β-Glu-RGD2 radiochemical purity.With the standard items for determining structure19F-AlF-NOTA-Glu、19F-AlF-NOTA-NOC and19F-AlF-NOTA-PEG3- β-Glu-RGD2, respectively with it is corresponding18F- AlF-NOTA-Glu、18F-AlF-NOTA-NOC and18F-AlF-NOTA-PEG3- β-Glu-RGD2 parenteral solutions are together expelled to In HPLC, or together point sample row TLC, to determine whether its retention time or Rf value Rf are consistent, and confirmation prepares parenteral solution Authenticity, after measured its radiochemical purity be all higher than 95% (Fig. 3 a- Fig. 3 f).
HPLC analysis conditions：Analytical column is ZORBAX Eclipse XDB-C18 posts, and mobile phase is 0.1%TFA acetonitrile Solution：The 0.1%TFA aqueous solution, row gradient elution：During 0min, acetonitrile solution/0.1%TFA's containing 0.1%TFA is water-soluble Liquid：2/98；When being gradually raised to 8min, the 0.1%TFA acetonitrile solution/0.1%TFA aqueous solution：10/90；It is raised again to 20min When, the 0.1%TFA acetonitrile solution/0.1%TFA aqueous solution：80/20.Flow velocity is 1mL/min, ultraviolet detection wavelength 210nm And 254nm.
TLC methods detect18F-AlF-NOTA-Glu、18F-AlF-NOTA-NOC and18F-AlF-NOTA-PEG3-β-Glu- The radiochemical purity of RGD2 parenteral solutions.A silica gel plate is taken, is put into behind shielding lead glass, a little radiation is drawn with capillary Property sample and its standard items (concentration 0.5mg/mL), together gently put on silica gel plate at one 1.5cm of distance, with hair dryer handle It is dried up.Chromatographed in chromatography cylinder, solvent is methanol：1.0M ammonium acetate=50:50 (V/V), hot-air is used after chromatography Drying, using radioactivity TLC scanner row thin layer scannings.After scanning, with iodine staining TLC plates, radioactive sample and standard are detected The Rf value Rf of product.
The lotus knurl model small animal position emission tomography (PET) of embodiment 5 images
Lotus prostate cancer nude mice model or normal nude mice, every group 3, injected respectively by tail vein18F-AlF-NOTA-Glu 、18F-AlF-NOTA-NOC and18F-AlF-NOTA-PEG3- β-Glu-RGD2 parenteral solutions (0.2mL, about 3.7-5.5MBq) are extremely naked In mouse model body.10min is placed on fixed plate gluing through 5% chloraldurate (6mL/kg) anesthesia nude mice is injected intraperitoneally before imaging Band is fixed, and body temperature is kept with heating cushion.After CT scan, PET data is collected in injection developer different time points, uses software After (Inevon Research Workplace 4.1) row correction for attenuation, iterative approximation image.Sketch the contours of tumour and brain, muscle Deng the area-of-interest (ROIs) of tissue, radiocounting and the volume of region of interest tissue are measured, calculates per gram of tissue injection Dosage percent (%ID/g), and calculate the radioactive uptake relative ratios such as tumour/brain, tumour/muscle.Representative PET imagings As a result (Fig. 4 a- Fig. 4 c) is shown：18F-AlF-NOTA-PEG3- β-Glu-RGD2 have in bilateral tumor tissues, kidney and bladder Higher intake,18F-AlF-NOTA-Glu has higher intake in kidney and bladder,18F-AlF-NOTA-NOC is in enteron aisle, kidney And have higher intake in bladder.
It is provided in an embodiment of the present invention18F labeled amino acid polypeptide drug synthetic methods, are produced by cyclotron 's18F-After being trapped by anion QMA pillars, eluted with a small amount of physiological saline (0.4-0.5mL)18F-, the whole of elution18F-Adopt With18It is prepared by F-AlF methods18F-AlF-NOTA- bioactive molecules, the method be referred to as QMA trapping-18F-AlF methods.QMA trappings-18F- AlF methods can not only complete full-automatic radiation synthesis, and can realize quick, efficient, high dose industry metaplasia using kit Production18F-AlF-NOTA- bioactive molecules, it is18The GMP productions of F- mark bioactive molecules and clinical Transformation Application are established Basis, can fully meet market and growing number of patient P ET checks needs.QMA trappings-18The Transformation Application of F-AlF methods will Open up18F- mark bioactive molecule research and development and the clinical Transformation Application new situation, promote the development of accurate diagnosis and treatment, and will produce huge Big social benefit and social benefit.
Above-described embodiment, the purpose of the present invention, technical scheme and beneficial effect are carried out further Describe in detail, should be understood that the embodiment that the foregoing is only the present invention, be not intended to limit the present invention Protection domain, within the spirit and principles of the invention, any modification, equivalent substitution and improvements done etc., all should include Within protection scope of the present invention.

Claims (10)

  1. It is 1. a kind of18F labeled amino acid polypeptide drug synthetic methods, it is characterised in that methods described includes：
    The capture cyclotron production of anion pillar18F-Ion；
    0.4-0.5ml physiological saline or other inorganic salt solutions after anion pillar by eluting18F-Ion obtains first Reaction liquid, into reaction bulb；
    By precursor-AlCl of the pH value between 4-53Acid solution-organic solvent mixing liquid adds reaction bulb；The precursor is NOTA derivatives；The NOTA is specially the alkyl triacetic acid base of three azacyclo- nine or the alkyl diacetoxyl of three azacyclo- nine；
    Synthesis is marked to 100 DEG C -110 DEG C in heating response bottle；
    After cooling, liquid in the reaction bulb is diluted with water；
    Liquid after being diluted in the reaction bulb is separated by solid phase extraction column so that product absorption to be obtained is solid Mutually on extraction pillar；
    Solid phase extraction column is cleaned with water, removes organic solvent and impurity；
    With ethanol rinse solid phase extraction column, product to be obtained is washed into product bottle, and add preparation liquid to obtain synthesized product.
  2. It is 2. according to claim 118F labeled amino acid polypeptide drug synthetic methods, it is characterised in that the anion Pillar is specially SEP-PAK Light QMA pillars.
  3. It is 3. according to claim 118F labeled amino acid polypeptide drug synthetic methods, it is characterised in that the precursor- AlCl3Than for 1.2:1.0, the quality of the precursor is 200-300 μ g.
  4. It is 4. according to claim 118F labeled amino acid polypeptide drug synthetic methods, it is characterised in that the solid phase extraction It is specially C18 pillars or HLB pillars to take pillar.
  5. It is 5. according to claim 118F labeled amino acid polypeptide drug synthetic methods, it is characterised in that the precursor is The alkyl diacetoxyl of three azacyclo- nine-Pidolidone NOTA-Glu, first reaction liquid are18F-AlF, the product are18The alkyl diacetoxyl of tri- azacyclo-s of F- nine-Pidolidone18F-AlF-NOTA-Glu。
  6. It is 6. according to claim 118F labeled amino acid polypeptide drug synthetic methods, it is characterised in that the precursor is Alkyl diacetoxyl -1- the naphthylalanines of three azacyclo- nine-Octreotide NOTA-NOC, first reaction liquid are18F-AlF, institute Stating product is18Alkyl diacetoxyl -1- the naphthylalanines of tri- azacyclo-s of F- nine-Octreotide18F-AlF-NOTA-NOC。
  7. It is 7. according to claim 118F labeled amino acid polypeptide drug synthetic methods, it is characterised in that the precursor is Polyethylenimine base-the β of three azacyclo-s, nine alkyl triacetic acid base-three-glutamic acid base-cyclic divalent arginine-glycine-aspartic acid NOTA-PEG3- β-Glu-RGD2, first reaction liquid are18F-AlF, the product are18The alkyl three of tri- azacyclo-s of F- nine The Polyethylenimine of acetate-three base-cyclic divalent arginine-glycine-aspartic acid18F-AlF-NOTA-PEG3-β-Glu- RGD2。
  8. It is 8. any described according to claim 1-718F labeled amino acid polypeptide drug synthetic methods, it is characterised in that institute State18F labeled amino acid polypeptide drug synthetic methods are used for18The full-automatic industrialization production of F-AlF-NOTA- biomolecule.
  9. 9. one kind is used for18The kit of F labeled amino acid polypeptide drug synthetic methods, it is characterised in that the kit bag Include：The first reagent bottle equipped with 0.4ml-0.5ml physiological saline, equipped with 0.4ml-1.2ml precursors-AlCl3The second reagent bottle With sub- reagent bottle, the 3rd reagent bottle equipped with 15ml water, the 4th reagent equipped with 10ml water that pH is 4-5 acidity acetonitrile solutions Bottle, the 5th reagent bottle equipped with 0.5-1.5ml ethanol, the 6th of physiological saline equipped with 10-18ml ascorbic acid containing 0.57mM the Reagent bottle, anion pillar, solid phase extraction column, anion pillar, prepared microcolumn solvent or solution, syringe, syringe needle, connect Receive bottle and inlet and outlet sterilised membrane filter.
  10. 10. kit according to claim 9, it is characterised in that the anion pillar is specially SEP-PAK Light QMA pillars；The solid phase extraction column is specially C18 pillars or HLB pillars；The prepared microcolumn solvent or solution are specific Including ethanol, water for injection and sodium bicarbonate solution.
CN201710821964.6A 2017-09-13 2017-09-13 Method and kit for synthesizing 18F-labeled amino acid polypeptide drug Active CN107501393B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710821964.6A CN107501393B (en) 2017-09-13 2017-09-13 Method and kit for synthesizing 18F-labeled amino acid polypeptide drug

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710821964.6A CN107501393B (en) 2017-09-13 2017-09-13 Method and kit for synthesizing 18F-labeled amino acid polypeptide drug

Publications (2)

Publication Number Publication Date
CN107501393A true CN107501393A (en) 2017-12-22
CN107501393B CN107501393B (en) 2021-04-09

Family

ID=60696576

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710821964.6A Active CN107501393B (en) 2017-09-13 2017-09-13 Method and kit for synthesizing 18F-labeled amino acid polypeptide drug

Country Status (1)

Country Link
CN (1) CN107501393B (en)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107970458A (en) * 2018-01-03 2018-05-01 山东省肿瘤防治研究院（山东省肿瘤医院） A kind of positron emitting tracer Fully automated synthesis module based on 18F-BF3 marks
CN108144073A (en) * 2017-12-28 2018-06-12 中山大学附属第医院 Target the three polyethyleneglycol modified duramycin polypeptide drugs of radioactive label of phosphatidyl-ethanolamine
CN110128504A (en) * 2019-04-29 2019-08-16 南方医科大学南方医院 [18F] aluminum fluoride and NOTA labelling method automatic production method
CN110551075A (en) * 2019-08-06 2019-12-10 唐刚华 Radiolabeled sub-glutamate compound and application thereof in preparation of PET (polyethylene terephthalate) imaging agent
CN112028914A (en) * 2020-08-14 2020-12-04 北京大学 A kind of18F-boron trifluoride tyrosine kit and application thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102391168A (en) * 2011-09-16 2012-03-28 中山大学附属第一医院 Symmetrical difunctional coupling agent and coupled molecular imaging agents thereof
CN104262073A (en) * 2014-09-30 2015-01-07 北京善为正子医药技术有限公司 Small modular multifunctional automatic 18F labelling PET (positron emission tomography) drug synthesizer

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102391168A (en) * 2011-09-16 2012-03-28 中山大学附属第一医院 Symmetrical difunctional coupling agent and coupled molecular imaging agents thereof
CN104262073A (en) * 2014-09-30 2015-01-07 北京善为正子医药技术有限公司 Small modular multifunctional automatic 18F labelling PET (positron emission tomography) drug synthesizer

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108144073A (en) * 2017-12-28 2018-06-12 中山大学附属第医院 Target the three polyethyleneglycol modified duramycin polypeptide drugs of radioactive label of phosphatidyl-ethanolamine
CN107970458A (en) * 2018-01-03 2018-05-01 山东省肿瘤防治研究院（山东省肿瘤医院） A kind of positron emitting tracer Fully automated synthesis module based on 18F-BF3 marks
CN110128504A (en) * 2019-04-29 2019-08-16 南方医科大学南方医院 [18F] aluminum fluoride and NOTA labelling method automatic production method
CN110128504B (en) * 2019-04-29 2023-08-22 南方医科大学南方医院 [ 18 F]Automatic production method for aluminum fluoride and NOTA labeling method
CN110551075A (en) * 2019-08-06 2019-12-10 唐刚华 Radiolabeled sub-glutamate compound and application thereof in preparation of PET (polyethylene terephthalate) imaging agent
CN112028914A (en) * 2020-08-14 2020-12-04 北京大学 A kind of18F-boron trifluoride tyrosine kit and application thereof
CN112028914B (en) * 2020-08-14 2021-11-16 北京大学 A kind of18F-boron trifluoride tyrosine kit and application thereof

Also Published As

Publication number Publication date
CN107501393B (en) 2021-04-09

Similar Documents

Publication Publication Date Title
CN107501393A (en) 18F labeled amino acid polypeptide drug synthetic methods and kit
Park et al. Luminescence imaging using radionuclides: a potential application in molecular imaging
CN106581700B (en) A kind of novel polypeptide radiopharmaceutical for targeting HER2 and its preparation method and application
CN105126128B (en) A kind of tumour VEGFR-3 molecular imagings agent and its application
CN104725473B (en) A kind of [18F] AlF marks PET polypeptide probes and preparation method thereof
CN107353323A (en) Al18PSMA targeted inhibition agent of F marks and preparation method and application
CN103830753A (en) Imaging drug <68>Ga-NOTA-IF7 targeting Anxa1 in tumor blood vessels and preparation method thereof
CN108187077A (en) 64PSMA targeted inhibition agent of Cu labels and preparation method and application
CN106415734A (en) Production of 43Sc radionuclide and radiopharmaceuticals thereof for use in positron emission tomography
CN106902363B (en) Radioactive composition, its single fraction preparation method and use
CN107311941A (en) 18EGFR positive electron tracers of F marks and preparation method and application
CN110339375A (en) A kind of rk polypeptide radiopharmaceutical and preparation method thereof targeting HER2
CN108434469A (en) A kind of HER2 affinities body68Ga markers and preparation method thereof, application
CN105713075A (en) EphB4 acceptor targeting polypeptide and applications thereof
CN107118097A (en) 2-18F- fluoropropionic acids isomers, its synthetic method and its application
Chakravarty et al. A simple and robust method for radiochemical separation of no-carrier-added 64Cu produced in a research reactor for radiopharmaceutical preparation
CN113105432B (en) Carbon-11 (C)11C) Radiopharmaceutical, preparation method and application thereof
CN105884867B (en) 18The affinity body class compound and the preparation method and application thereof of F label
CN102452873B (en) Compound containing carbon or oxygen isotope and preparation method thereof and application thereof and composition thereof
CN115286693A (en) Tumor targeting peptide aiming at carcinoembryonic antigen related cell adhesion molecule CEACAM6 and application thereof
Gómez‐Vallejo et al. Synthesis and in vivo evaluation of 11C‐labeled (1, 7‐dicarba‐closo‐dodecaboran‐1‐yl)‐N‐{[(2S)‐1‐ethylpyrrolidin‐2‐yl] methyl} amide
CN107021998A (en) A kind of positron radionuclide labeling polypeptide for tumor imaging
CN107847617A (en) Metal tricarbonyl compound containing substituted iminodiacetic acid part and the purposes as radioisotopic tracer
CN114853851B (en) Targeting PD-L1 polypeptide probe and application thereof in preparation of PET imaging agent
CN112028914B (en) A kind of18F-boron trifluoride tyrosine kit and application thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant