CN107501393B

CN107501393B - Method and kit for synthesizing 18F-labeled amino acid polypeptide drug

Info

Publication number: CN107501393B
Application number: CN201710821964.6A
Authority: CN
Inventors: 唐刚华; 胡晓平; 胡生焰
Original assignee: Beijing Pet Co ltd
Current assignee: Beijing Pet Co ltd
Priority date: 2017-09-13
Filing date: 2017-09-13
Publication date: 2021-04-09
Anticipated expiration: 2037-09-13
Also published as: CN107501393A

Abstract

Practice of the inventionExample relates to a¹⁸A method and a kit for synthesizing F-labeled amino acid polypeptide drugs are provided, wherein the method comprises the following steps: produced by an anion small column capture cyclotron¹⁸F^‑Ions; 0.4-0.5ml of physiological saline is eluted after passing through the anion small column¹⁸F^‑Obtaining first reaction liquid by ions, and feeding the first reaction liquid into a reaction bottle; precursor-AlCl with pH value between 4 and 5₃Adding the acid solution-organic solvent mixed liquid into a reaction bottle; the precursor is a NOTA derivative; NOTA is triazacyclononalkyltriacetoxy or triazacyclononaalkyldiacetoxy; heating the reaction bottle to 100-110 ℃ for labeling synthesis; after cooling, adding water to dilute the liquid in the solid-phase extraction reaction bottle; separating the diluted liquid in the solid-phase extraction reaction bottle through a small solid-phase extraction column to enable the product to be obtained to be adsorbed on the small solid-phase extraction column; washing the solid phase extraction column with water to remove organic solvent and impurities; and leaching the solid-phase extraction small column by using ethanol, washing a product to be obtained into a product bottle, and adding the preparation solution to obtain the synthesized product.

Description

Method and kit for synthesizing 18F-labeled amino acid polypeptide drug

Technical Field

The invention relates to the field of drug synthesis, in particular to a compound¹⁸A method for synthesizing F-labeled amino acid polypeptide drugs and a kit.

Background

¹⁸F-fluorodeoxyglucose (F-fluorodeoxyglucose)¹⁸F-FDG) has been widely used for early diagnosis and treatment guidance of tumors, but is a non-specific positron emission tomography (PET drug, which is prone to false positive and false negative results.¹⁸F-labeled amino acids and targeting receptor specificity¹⁸F-labeled polypeptide PET medicine can compensate¹⁸The serious shortage of F-FDG is an important development direction for developing PET drugs. However, the existing general¹⁸The method for labeling the biomolecule by using N-succinimide-4-¹⁸F-fluorobenzoic acid ester (¹⁸F-SFB) and¹⁸F-4-Nitrophenyl-2-fluoropropionate (A)¹⁸F-NFP) to mark amino acid polypeptide-NH 2 group, and has the advantages of multiple reaction steps, complex operation, long synthesis time and difficult realization of automatic synthesis. Simple and practical Al¹⁸F labeling method¹⁸F is used for marking amino group of amino acid polypeptide drug.

With marking prosthetic groups¹⁸F-SFB and¹⁸preparation of F-NFP¹⁸F-labeled bioactive molecule amino is the most common method, but the method is complex, has low radiochemical yield and is difficult to complete industrial automatic production. B-¹⁸F or Si-¹⁸F one-step method can efficiently and automatically prepare¹⁸F-labelled biologically active molecules, but obtained by this method are vectors containing¹⁸F-labeled amino acid polypeptide drugs.

¹⁸The F-AlF method has been used to prepare targeted somatostatin receptors¹⁸F-triazacyclononaalkyldiacetoxy-1-naphthylalanine-octreotide (¹⁸F-AlF-NOTA-NOC) and targeting integrin α v β 3¹⁸F-triazacyclononaalkyltriacetoxy-trethylolamino-cyclic divalent arginine-glycine-aspartic acid ((¹⁸F-AlF-NOTA-PEG3-Glu-RGD2)，¹⁸F-AlF-NOTA-NOC and¹⁸F-AlF-NOTA-PEG3-Glu-RGD2 shows better clinical application prospect. Recently, we have also developed targeting integrin α v β 3¹⁸F-labeled beta-glutamic acid coupled bivalent symmetry¹⁸F-AlF-NOTA-PEG 3-beta-Glu-RGD 2 and¹⁸F-triazacyclononaalkyldiacetic-L-glutamic acid (a)¹⁸F-AlF-NOTA-Glu) and is applied to PET imaging research of a plurality of tumor-bearing model mice, showing better application potential. However, there are existing¹⁸F-AlF labeled amino acid polypeptide drugs, e.g.¹⁸F-AlF-NOTA-Glu、¹⁸F-AlF-NOTA-NOC、¹⁸F-AlF-NOTA-PEG3-Glu-RGD2 and¹⁸F-AlF-NOTA-PEG 3-beta-Glu-RGD 2 and the like, and only a kit method is adopted to prepare low dose¹⁸F-AlF-NOTA-biomolecules, not all that could be produced by the accelerator¹⁸F^-Direct application of ions to¹⁸F-AlF marks amino acid and polypeptide, is difficult to realize full-automatic industrial production, and brings certain risk to radiation safety protection.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides a novel device¹⁸A method for synthesizing F-labeled amino acid polypeptide drugs and a kit.

In view of the above, in a first aspect, embodiments of the present invention provide a method¹⁸F-labeled amino acidThe method for synthesizing the polypeptide drug comprises the following steps:

produced by an anion small column capture cyclotron¹⁸F^-Ions;

0.4-0.5ml physiological saline or other inorganic saline solution is eluted after passing through anion column¹⁸F^-Obtaining first reaction liquid by ions, and feeding the first reaction liquid into a reaction bottle;

precursor-AlCl with pH value between 4 and 5₃Adding the acid solution-organic solvent mixed liquid into a reaction bottle; the precursor is a NOTA derivative; the NOTA is specifically triazacyclononaalkyltriacetic acid or triazacyclononaalkyldiacetic acid;

heating the reaction bottle to 100-110 ℃ for labeling synthesis;

after cooling, adding water to dilute the liquid in the reaction bottle;

separating the diluted liquid in the reaction bottle through a solid phase extraction column to enable the product to be obtained to be adsorbed on the solid phase extraction column;

washing the solid phase extraction column with water to remove organic solvent and impurities;

and leaching the solid-phase extraction small column by using ethanol, washing a product to be obtained into a product bottle, and adding the preparation solution to obtain the synthesized product.

Preferably, the anion column is specifically an SEP-PAK Light QMA column.

Preferably, the precursor-AlCl₃The ratio was 1.2:1.0 and the mass of the precursor was 200-300. mu.g.

Preferably, the solid phase extraction column is a C18 column or an HLB column.

Preferably, the precursor is triazacyclononaalkyldiacetic-L-glutamic acid NOTA-Glu, and the first reaction liquid is¹⁸F-AlF, the product being¹⁸F-triazacyclononaalkyldiacetic acid-L-glutamic acid¹⁸F-AlF-NOTA-Glu。

Preferably, the precursor is triazacyclononaalkyldiacetic-1-naphthylalanine-octreotide NOTA-NOC, and the first reaction liquid is¹⁸F-AlF, the product being¹⁸F-triazaCyclononaalkyldiacetic-1-naphthylalanine-octreotide¹⁸F-AlF-NOTA-NOC。

Preferably, the precursor is triazacyclononaalkyltriacetoxy-trethylolamido-beta-glutamyl-cyclic divalent arginine-glycine-aspartic acid NOTA-PEG 3-beta-Glu-RGD 2, and the first reaction liquid is Triazacyclononaalkyltriacetoxy-trethylolamido-beta-glutamyl-cyclic divalent arginine-glycine-aspartic acid NOTA-PEG 3-beta-Glu-RGD 2¹⁸F-AlF, the product being¹⁸F-triazacyclononaalkyltriacetoxy-trethylolamino-cyclic divalent arginine-glycine-aspartic acid¹⁸F-AlF-NOTA-PEG3-β-Glu-RGD2。

Preferably, the¹⁸Method for synthesizing F-labeled amino acid polypeptide drug¹⁸And (3) full-automatic industrial production of the F-AlF-NOTA-biomolecule.

In a second aspect, the embodiments of the present invention provide a method for¹⁸The kit of the F-labeled amino acid polypeptide drug synthesis method comprises the following steps: a first reagent bottle containing 0.4ml to 0.5ml of physiological saline, a second reagent bottle containing 0.4ml to 1.2ml of precursor AlCl3 and a sub-reagent bottle with pH of 4 to 5 acidic acetonitrile solution, a third reagent bottle containing 15ml of water, a fourth reagent bottle containing 10ml of water, a fifth reagent bottle containing 0.5 to 1.5ml of ethanol, a sixth reagent bottle containing 10 to 18ml of physiological saline containing 0.57mM ascorbic acid, an anion column, a solid phase extraction column, an anion column, a pretreatment column solvent or solution, a syringe, a needle, a receiving bottle and an inlet and outlet sterile filter membrane.

Preferably, the anion column is a SEP-PAK Light QMA column; the solid phase extraction small column is specifically a C18 small column or an HLB small column; the pretreatment column solvent or solution specifically comprises ethanol, water for injection and sodium bicarbonate solution.

The embodiment of the invention provides¹⁸Synthesis method of F-labeled amino acid polypeptide drug and production method of F-labeled amino acid polypeptide drug by cyclotron¹⁸F^-After being trapped by the anion QMA small column, the gel is eluted by a small amount of physiological saline¹⁸F^-All of the elution¹⁸F^-By using¹⁸Preparation by F-AlF method¹⁸F-AlF-NOTA-bioactive molecules, a method called QMA Capture-¹⁸The F-AlF method. QMA Capture-¹⁸F-AlFThe method can not only complete full-automatic radiosynthesis, but also realize rapid, high-efficiency and high-dose industrialized production by adopting the kit¹⁸F-AlF-NOTA-bioactive molecule of the formula¹⁸The GMP production and clinical transformation application of the F-labeled bioactive molecules lay a foundation, and can completely meet the requirements of the market and the PET examination of the patients who are continuously increased. QMA Capture-¹⁸The conversion application of the F-AlF method will be opened up¹⁸The F-labeled bioactive molecule is applied in research and development and clinical transformation, promotes the development of accurate diagnosis and treatment, and generates great social benefit and social benefit.

Drawings

FIG. 1 is a schematic diagram of an embodiment of the present invention¹⁸F marks the technological process schematic diagram of the amino acid polypeptide drug synthesis;

FIG. 2a is a schematic diagram of an embodiment of the present invention¹⁸A schematic diagram of the synthetic process of F-AlF-NOTA-Glu;

FIG. 2b is a schematic diagram of an embodiment of the present invention¹⁸A schematic diagram of the synthetic process of F-AlF-NOTA-NOC;

FIG. 2c is a schematic diagram of an embodiment of the present invention¹⁸A schematic diagram of the synthetic process of F-AlF-NOTA-PEG 3-beta-Glu-RGD 2;

FIG. 3a is a schematic diagram of an embodiment of the present invention¹⁸High Performance Liquid Chromatography (HPLC) analysis profile of F-AlF-NOTA-NOC;

FIG. 3b is a schematic diagram of an embodiment of the present invention¹⁸A Thin Layer Chromatography (TLC) analysis profile of F-AlF-NOTA-NOC;

FIG. 3c is a schematic diagram of an embodiment of the present invention¹⁸An HPLC analysis profile of F-AlF-NOTA-PEG 3-beta-Glu-RGD 2;

FIG. 3d is a schematic diagram of an embodiment of the present invention¹⁸A TLC analysis map of F-AlF-NOTA-PEG 3-beta-Glu-RGD 2;

FIG. 3e is a schematic diagram of an embodiment of the present invention¹⁸HPLC analysis pattern of F-AlF-NOTA-Glu;

FIG. 3f is a schematic diagram of an embodiment of the present invention¹⁸TLC analysis pattern of F-AlF-NOTA-Glu.

FIG. 4A is a view showing a dual tumor-bearing model according to an embodiment of the present invention¹⁸F-AlF-NOTA-PEG 3-beta-Glu-RGD 2 positron emission tomography PET image

FIG. 4B shows a normal animal according to an embodiment of the present invention¹⁸PET images of F-AlF-NOTA-NOC;

FIG. 4C shows a normal animal according to the present invention¹⁸PET images of F-AlF-NOTA-Glu.

Detailed Description

The technical solution of the present invention is further described in detail by the accompanying drawings and embodiments.

The process flow diagram of the method for synthesizing the 18F-labeled amino acid polypeptide drug provided by the embodiment of the invention is shown in figure 1. Production by cyclotron¹⁸F^-Ions, trapped by anionic columns (identified as QMA in the figure)¹⁸F^-Ions; then 0.4-0.5ml of physiological saline is passed through the anion column for elution¹⁸F^-Ions are obtained to obtain a first reaction liquid, and the first reaction liquid enters a reaction bottle 10; and, the precursor-AlCl with pH value between 4 and 5₃Adding the acid solution-organic solvent mixed liquid into a reaction bottle 10; wherein the precursor is a NOTA derivative; NOTA is triazacyclononalkyltriacetoxy or triazacyclononaalkyldiacetoxy; then heating the reaction bottle 10 to 100-110 ℃, carrying out a chelating reaction, and carrying out label synthesis; after cooling, adding water to dilute the liquid in the reaction bottle 10; separating the diluted liquid in the reaction bottle 10 by a solid phase extraction column (marked as C-18 in the figure) to ensure that the product to be obtained is adsorbed on the solid phase extraction column; washing the solid phase extraction column with water to remove organic solvent and impurities; and leaching the solid phase extraction small column by using ethanol, washing a product to be obtained into a product bottle 9, and adding the preparation solution to obtain the synthesized product.

Wherein, SEP-PAK Light QMA column can be used as anion column; precursor AlCl₃The ratio is 1.2:1.0, the mass of the precursor is 200-; the solid phase extraction column can specifically adopt a C18 column or an HLB column.

In a specific example, the precursor is triazacyclononaalkyldiacetic-L-glutamic acid (NOTA-Glu), and the first reaction liquid is¹⁸F-AlF, the product is¹⁸F-triazacyclononaalkyldiacetic-L-glutamic acid (a)¹⁸F-AlF-NOTA-Glu). The schematic diagram of the synthesis process is shown in fig. 2 a.

In another specific example, the precursor is triazacyclononaalkyldiacetic-1-naphthylalanine-octreotide (NOTA-NOC) and the first reaction liquid is¹⁸F-AlF, the product is¹⁸F-triazacyclononaalkyldiacetoxy-1-naphthylalanine-octreotide (¹⁸F-AlF-NOTA-NOC). The schematic diagram of the synthesis process is shown in fig. 2 b.

In yet another specific example, the precursor is triazacyclononanoalkyltriacetoxy-trethylolamino-beta-glutamyl-cyclic divalent arginine-glycine-aspartic acid NOTA-PEG 3-beta-Glu-RGD 2 and the first reaction liquid is Triazacyclononalkyltriacetoxy-trimerization-glycolic acid-beta-glutaminyl-cyclic divalent arginine-glycine-aspartic acid NOTA-PEG 3-beta-Glu-RGD 2¹⁸F-AlF, the product is¹⁸F-triazacyclononaalkyltriacetoxy-trethylolamino-cyclic divalent arginine-glycine-aspartic acid ((¹⁸F-AlF-NOTA-PEG 3-beta-Glu-RGD 2). The schematic of the synthesis is shown in FIG. 2 c.

Of the present embodiment¹⁸The method for synthesizing F-labeled amino acid polypeptide drugs can be used for¹⁸And (3) full-automatic industrial production of the F-AlF-NOTA-biomolecule.

To facilitate the implementation of the method, the invention also provides a kit comprising: a first reagent bottle containing 0.4ml to 0.5ml of physiological saline, a second reagent bottle containing 0.4ml to 1.2ml of precursor AlCl3 and a sub-reagent bottle with pH of 4 to 5 acidic acetonitrile solution, a third reagent bottle containing 15ml of water, a fourth reagent bottle containing 10ml of water, a fifth reagent bottle containing 0.5 to 1.5ml of ethanol, a sixth reagent bottle containing 10 to 18ml of physiological saline containing 0.57mM ascorbic acid, an anion column, a solid phase extraction column, an anion column, a pretreatment column solvent or solution, a syringe, a needle, a receiving bottle and an inlet and outlet sterile filter membrane.

Similarly, in the kit, the anion small column is specifically an SEP-PAK Light QMA small column; the solid phase extraction column is specifically a C18 column or an HLB column; the pre-treatment column solvent or solution specifically includes ethanol, water for injection and sodium bicarbonate solution.

Hereinafter, specific examples will be described in further detail.

Example 1¹⁸Full-automatic industrial synthesis of F-AlF-NOTA-Glu injection

The kit comprises the following components: 0.4-0.5mL of physiological saline (vial 1), 200-₃(bottle 2), 0.8-1.2mL of pH4-5 acidic acetonitrile solution (bottle 2 ') (bottle 2 ' is in the kit, but not shown in FIG. 1, the solution from bottle 2 ' was added to bottle 2 prior to synthesis), 15mL of water for injection (bottle 3), 10mL of water for injection (bottle 4), 1.5mL of ethanol (bottle 5), and 18mL of physiological saline containing 0.57mM ascorbic acid (bottle 6). The auxiliary material comprises: SEP-PAK C18 separation cartridge, SEP-PAK QMA cartridge, cartridge pre-treatment solvent or solution (ethanol, water for injection, 8.4% sodium bicarbonate solution), syringe and needle, receiving vial, inlet and outlet sterile filter. SEP-PAK QMA small column can be replaced by other anion small column; the physiological saline can be replaced by other inorganic saline solution; acetonitrile can be replaced by dimethyl sulfoxide, ethanol and methanol; SEP-PAK C18 columella, or HLB columella.

Design of¹⁸F-AlF mark automatic synthesis program, starting self-made small-sized¹⁸Before the F-FDG synthesizer (shown in figure 1), adding the acidic acetonitrile solution in the bottle 2' into the bottle 2; pretreating the SEP-PAK QMA small column with 8.4% sodium bicarbonate solution and water for injection in sequence; sequentially pretreating SEP-PAK C18 separation small column with ethanol and water for injection; vial 1, vial 2, vial 3, vial 4, vial 5, vial 6, SEP-PAK QMA cartridge, SEP-PAK C18 separation cartridge, and the receiving vial with inlet and outlet sterile filter were connected to the synthesizer (as shown in fig. 1) at the corresponding locations.

Starting the synthesizer to start production¹⁸F-AlF-NOTA-Glu injection. By a cyclotron¹⁸O(p,n)¹⁸Produced by reaction of F nuclei¹⁸F^-In N at₂Under the air-borne band, the mixture passes through a Sep-Pak QMA anion small column in a radioactivity activity meter,¹⁸the F-is trapped in the small column,¹⁸the O-water was collected in a recovery bottle. In N₂Under the belt, the physiological saline in the bottle 1 is put into the QMA small column¹⁸F^-The eluate is taken out into a closed reaction flask 10. Then, in N₂Under the carrier tape, in bottle 2, NOTA-Glu +2mM 24. mu.L AlCl₃The solution is added toMixing the mixture in a reaction bottle 10, and heating the mixture at the temperature of 100 ℃ and 110 ℃ for reaction for 10 min. After cooling, 15mL of water for injection in vial 3 was added to reaction vial 10, mixed well and transferred to a SEP-PAK C18 mini column. After the solution in the reaction flask had been completely transferred, the column was rinsed with 10mL of water for injection in flask 4, and N was added₂And (5) drying the small column. Finally, the ethanol in the bottle 5 elutes the product into a receiving bottle 8, and the normal saline in the bottle 6 washes the small column to dilute the product, so as to obtain the product meeting the requirements¹⁸F-AlF-NOTA-Glu injection.¹⁸F-AlF-NOTA-Glu uncorrected radiochemical yield is between 40% and 60%.

Example 2¹⁸Full-automatic industrial synthesis of F-AlF-NOTA-NOC injection

The kit comprises the following components: 0.4-0.5mL physiological saline (vial 1), 200-₃(vial 2), 0.8-1.2mL pH4-5 acidic acetonitrile solution (vial 2'), 15mL physiological saline (vial 3), 10mL physiological saline (vial 4), 1.5mL ethanol (vial 5), and 18mL physiological saline containing 0.57mM ascorbic acid (vial 6). The auxiliary material comprises: an HLB column, an SEP-PAK QMA column, a column pretreatment solvent or solution (ethanol, physiological saline, water for injection, 8.4% sodium bicarbonate solution), a syringe and needle, a receiving vial, and sterile filter membranes for inlet and outlet. SEP-PAK QMA small column can be replaced by other anion small column; the physiological saline can be replaced by other inorganic saline solution; acetonitrile can be replaced by dimethyl sulfoxide, ethanol and methanol; the HLB column can also be replaced by SEP-PAK C18 separation column.

Design of¹⁸F-AlF mark automatic synthesis program, starting self-made small-sized¹⁸Before the F-FDG synthesizer (shown in figure 1), adding the acidic acetonitrile solution in the bottle 2' into the bottle 2; pretreating the SEP-PAK QMA small column with 8.4% sodium bicarbonate solution and water for injection in sequence; pretreating the HLB column with ethanol and physiological saline in turn; vial 1, vial 2, vial 3, vial 4, vial 5, vial 6, SEP-PAK QMA cartridge, HLB cartridge, and receiving vial with inlet and outlet sterile filter were attached to the synthesizer (fig. 1) at the corresponding locations.

Starting the synthesizer to start production¹⁸F-AlF-NOTA-NOC injection. By a cyclotron¹⁸O(p,n)¹⁸Produced by reaction of F nuclei¹⁸F^-In N at₂Under the air-borne band, the mixture passes through a Sep-Pak QMA anion small column in a radioactivity activity meter,¹⁸F^-is trapped in the small column and is collected in the small column,¹⁸the O-water was collected in a recovery bottle. In N₂Under the belt, the physiological saline in the bottle 1 is put into the QMA small column¹⁸F^-The eluate is taken out into a closed reaction flask 10. Then, in N₂Under tape, in bottle 2 NOTA-NOC +2mM 24. mu.L AlCl₃The solution is added into a reaction bottle 10, mixed evenly and heated at the temperature of 100 ℃ and 110 ℃ for reaction for 10 min. After cooling, 15mL of physiological saline in vial 3 was added to reaction vial 10, mixed well, and transferred to an HLB column. After the solution in the reaction flask was completely transferred, the column was rinsed with 10mL of physiological saline in flask 4, and N was added₂And (5) drying the small column. Finally, the ethanol in the bottle 5 elutes the product into a receiving bottle 8, and the normal saline in the bottle 6 washes the small column to dilute the product, so as to obtain the product meeting the requirements¹⁸F-AlF-NOTA-NOC injection.¹⁸F-AlF-NOTA-NOC uncorrected radiochemical yield was 40% -60%.

Example 3¹⁸Full-automatic industrial synthesis of F-AlF-NOTA-PEG 3-beta-Glu-RGD 2 injection

The kit comprises the following components: 0.4-0.5mL of physiological saline (vial 1), 200-₃(bottle 2), 0.8-1.2mL pH4-5 acidic acetonitrile solution (bottle 2'), 15mL water for injection (bottle 3), 10mL water for injection (bottle 4), 1.5mL ethanol (bottle 5), and 18mL physiological saline containing 0.57mM ascorbic acid (bottle 6). The auxiliary material comprises: SEP-PAK C18 separation cartridge, SEP-PAK QMA cartridge, cartridge pre-treatment solvent or solution (ethanol, water for injection, 8.4% sodium bicarbonate solution), syringe and needle, receiving vial, inlet and outlet sterile filter. SEP-PAK QMA small column can be replaced by other anion small column; the physiological saline can be replaced by other inorganic saline solution; acetonitrile can be replaced by dimethyl sulfoxide, ethanol and methanol; SEP-PAK C18 columella, or HLB columella.

Design of¹⁸F-AlF mark automatic synthesis program, starting self-made small-sized¹⁸Before the F-FDG synthesizer (shown in figure 1), adding the acidic acetonitrile solution in the bottle 2' into the bottle 2; SEP-PAK QMA small column is sequentially injected with 8.4% sodium bicarbonate solutionPretreating with water; sequentially pretreating SEP-PAK C18 separation small column with ethanol and water for injection; vial 1, vial 2, vial 3, vial 4, vial 5, vial 6, SEP-PAK QMA cartridge, SEP-PAK C18 separation cartridge, and the receiving vial with inlet and outlet sterile filter were connected to the synthesizer (as shown in fig. 1) at the corresponding locations.

Starting the synthesizer to start production¹⁸F-AlF-NOTA-PEG 3-beta-Glu-RGD 2 injection. By a cyclotron¹⁸O(p,n)¹⁸Produced by reaction of F nuclei¹⁸F^-In N at₂Under the air-borne band, the mixture passes through a Sep-Pak QMA anion small column in a radioactivity activity meter,¹⁸F^-is trapped in the small column and is collected in the small column,¹⁸the O-water was collected in a recovery bottle. In N₂Under the belt, the physiological saline in the bottle 1 is put into the QMA small column¹⁸F^-The eluate is taken out into a closed reaction flask 10. Then, in N₂Under the belt, in the bottle 2, NOTA-PEG 3-beta-Glu-RGD 2+2mM 24 mu L AlCl₃The solution is added into a reaction bottle 10, mixed evenly and heated at the temperature of 100 ℃ and 110 ℃ for reaction for 10 min. After cooling, 15mL of water for injection in vial 3 was added to reaction vial 10, mixed well and transferred to a SEP-PAK C18 mini-column. After the solution in the reaction flask had been completely transferred, the column was rinsed with 10mL of water for injection in flask 4, and N was added₂And (5) drying the small column. Finally, the ethanol in the bottle 5 elutes the product into a receiving bottle 8, and the normal saline in the bottle 6 washes the small column to dilute the product, so as to obtain the product meeting the requirements¹⁸F-AlF-NOTA-PEG 3-beta-Glu-RGD 2 injection.¹⁸The uncorrected radiochemical yield of F-AlF-NOTA-PEG 3-beta-Glu-RGD 2 is 20-40%.

EXAMPLE 4 determination of product purity

Measuring by radioactive High Performance Liquid Chromatography (HPLC) and Thin Layer Chromatography (TLC)¹⁸F-AlF-NOTA-Glu、¹⁸F-AlF-NOTA-NOC and¹⁸radiochemical purity of F-AlF-NOTA-PEG 3-beta-Glu-RGD 2. By means of standards of defined construction¹⁹F-AlF-NOTA-Glu、¹⁹F-AlF-NOTA-NOC and¹⁹F-AlF-NOTA-PEG 3-beta-Glu-RGD 2, respectively corresponding to¹⁸F-AlF-NOTA-Glu、¹⁸F-AlF-NOTA-NOC and¹⁸F-AlF-NOTA-PEG 3-beta-Glu-RGD 2 injection is injected into HPLC together, or spotted together for TLC, so as toThe consistency of the retention time or the specific shift value Rf is determined and the authenticity of the injection preparation is confirmed, which is determined to have a radiochemical purity greater than 95% (FIGS. 3 a-3 f).

Conditions for HPLC analysis: the analytical column was a ZORBAX Eclipse XDB-C18 column, mobile phase 0.1% TFA in acetonitrile: aqueous 0.1% TFA, gradient elution: 0min, acetonitrile/0.1% TFA in water: 2/98, respectively; gradually increasing to 8min, 0.1% TFA in acetonitrile/0.1% TFA in water: 10/90, respectively; at 20min, 0.1% TFA in acetonitrile/0.1% TFA in water: 80/20. The flow rate was 1mL/min, and the UV detection wavelengths were 210nm and 254 nm.

Detection by TLC method¹⁸F-AlF-NOTA-Glu、¹⁸F-AlF-NOTA-NOC and¹⁸radiochemical purity of the F-AlF-NOTA-PEG 3-beta-Glu-RGD 2 injection. Taking a silica gel plate, placing behind the shielding lead glass, sucking a small amount of radioactive sample and a standard substance (with the concentration of 0.5mg/mL) thereof by using a capillary tube, lightly dropping the sample and the standard substance on the silica gel plate at a position 1.5cm away from one end of the silica gel plate, and drying the sample by using an electric blower. Carrying out chromatography in a chromatographic cylinder, wherein a developing solvent is methanol: 1.0M ammonium acetate 50:50(V/V), dried with hot air after chromatography and scanned in thin layers using a radioactive TLC scanner. After scanning, TLC plates were stained with iodine and the shift values Rf of radioactive samples and standards were determined.

Example 5 tumor-bearing model PET imaging of small animals

Nude mice model with prostate cancer or normal nude mice, 3 mice in each group, are injected from tail vein¹⁸F-AlF-NOTA-Glu、¹⁸F-AlF-NOTA-NOC and¹⁸F-AlF-NOTA-PEG3- β -Glu-RGD2 injection (0.2mL, about 3.7-5.5MBq) into nude mouse model. Nude mice were anesthetized by intraperitoneal injection of 5% chloral hydrate (6mL/kg) 10min before imaging, fixed on a fixing plate with an adhesive tape, and kept at body temperature with a heating pad. After CT scanning, PET data were collected at different time points with injected imaging agent, and after attenuation correction with software (Inevon Research Workplace 4.1), images were reconstructed iteratively. Delineating regions of interest (ROIs) of tumors and tissues such as brain, muscle, etc., measuring radioactivity counts and volumes of the tissues of interest, calculating percent injected dose per gram tissue (% ID/g), and calculating tumor @Relative ratio of radioactive uptake by brain, tumor/muscle, etc. Representative PET imaging results (fig. 4 a-4 c) show:¹⁸F-AlF-NOTA-PEG 3-beta-Glu-RGD 2 has higher uptake in bilateral tumor tissues, kidney and bladder,¹⁸F-AlF-NOTA-Glu has higher uptake in the kidney and bladder,¹⁸F-AlF-NOTA-NOC has a higher uptake in the intestine, kidney and bladder.

The embodiment of the invention provides¹⁸Method for synthesizing F-labeled amino acid polypeptide drug by cyclotron production¹⁸F^-After being trapped by the anion QMA small column, the column was eluted with a small amount of physiological saline (0.4-0.5mL)¹⁸F^-All of the elution¹⁸F^-By using¹⁸Preparation by F-AlF method¹⁸F-AlF-NOTA-bioactive molecules, a method called QMA Capture-¹⁸The F-AlF method. QMA Capture-¹⁸The F-AlF method can not only complete full-automatic radiosynthesis, but also realize rapid, high-efficiency and high-dose industrial production by adopting the kit¹⁸F-AlF-NOTA-bioactive molecule of the formula¹⁸The GMP production and clinical transformation application of the F-labeled bioactive molecules lay a foundation, and can completely meet the requirements of the market and the PET examination of the patients who are continuously increased. QMA Capture-¹⁸The conversion application of the F-AlF method will be opened up¹⁸The F-labeled bioactive molecule is applied in research and development and clinical transformation, promotes the development of accurate diagnosis and treatment, and generates great social benefit and social benefit.

The above-mentioned embodiments are intended to illustrate the objects, technical solutions and advantages of the present invention in further detail, and it should be understood that the above-mentioned embodiments are merely exemplary embodiments of the present invention, and are not intended to limit the scope of the present invention, and any modifications, equivalent substitutions, improvements and the like made within the spirit and principle of the present invention should be included in the scope of the present invention.

Claims

1. A kind of¹⁸The method for synthesizing the F-labeled amino acid polypeptide drug is characterized by comprising the following steps of:

produced by an anion small column capture cyclotron¹⁸F^-Ions; wherein the anion column is SEP-PAK Light QMA column;

0.4-0.5ml of physiological saline is eluted after passing through the anion small column¹⁸F^-Obtaining first reaction liquid by ions, and feeding the first reaction liquid into a reaction bottle;

precursor-AlCl with pH value between 4 and 5₃Adding the acid solution-organic solvent mixed liquid into a reaction bottle; the precursor is a NOTA derivative; the NOTA is specifically 1,4, 7-triazacycloalkane-1, 4, 7-triacetic acid-2-S- (4-isothiocyanatobenzyl) or 1,4, 7-triazacyclononaalkyl-N ', N' -diacetoxy-N-acetyl;

heating the reaction bottle to 100-110 ℃ for labeling synthesis;

after cooling, adding water to dilute the liquid in the reaction bottle;

separating the diluted liquid in the reaction bottle through a solid phase extraction column to enable the product to be obtained to be adsorbed on the solid phase extraction column; wherein the solid phase extraction column is specifically a C18 column;

leaching the solid-phase extraction small column by using ethanol, washing a product to be obtained into a product bottle, and adding the preparation solution to obtain a synthesized product;

wherein the precursor is 1,4, 7-triazacycloalkane-1, 4, 7-triacetic acid-2-S- (4-isothiocyanatobenzyl) -L-glutamic acid, and the product is⁸F-AlF-1,4, 7-triazacycloalkane-1, 4, 7-triacetic acid-2-S- (4-isothiocyanatobenzyl) -L-glutamic acid; or

The precursor is 1,4, 7-triazacyclononaalkyl-N ', N' -diacetoxy-N-acetyl-octreotide, and the product is¹⁸F-AlF-1,4, 7-triazacyclononaalkyl-N ', N' -diacetoxy-N-acetyl-octreotide;

the above-mentioned¹⁸Method for synthesizing F-labeled amino acid polypeptide drug¹⁸And (3) full-automatic industrial production of the F-AlF-NOTA-biomolecule.

2. The method of claim 1¹⁸F-labeled amino acid polypeptide drugMethod for synthesizing a compound, characterized in that the precursor-AlCl₃The ratio was 1.2:1.0 and the mass of the precursor was 200-300. mu.g.

3. Is used for¹⁸The kit for the synthesis method of the F-labeled amino acid polypeptide drug is characterized by comprising the following steps: a first reagent bottle containing 0.4ml-0.5ml of normal saline, and a first reagent bottle containing 0.4ml-1.2ml of AlCl as a precursor₃The second reagent bottle, a sub reagent bottle of an acidic acetonitrile solution with the pH value of 4-5, a third reagent bottle filled with 15ml of water, a fourth reagent bottle filled with 10ml of water, a fifth reagent bottle filled with 0.5-1.5ml of ethanol, a sixth reagent bottle filled with 10-18ml of physiological saline containing 0.57mM of ascorbic acid, an anion column, a solid phase extraction column, a pretreatment column solvent or solution, a syringe, a needle, a receiving bottle and an inlet and outlet sterile filter membrane;

wherein the anion column is SEP-PAK Light QMA column; the solid phase extraction small column is specifically a C18 small column;

the precursor is 1,4, 7-triazacycloalkane-1, 4, 7-triacetic acid-2-S- (4-isothiocyanatobenzyl) -L-glutamic acid; or

The precursor is 1,4, 7-triazacyclononaalkyl-N ', N' -diacetoxy-N-acetyl-octreotide;

the kit is used for¹⁸And (3) full-automatic industrial production of the F-AlF-NOTA-biomolecule.

4. Kit according to claim 3, characterized in that the pre-treatment column solvent or solution comprises in particular ethanol, water for injection and sodium bicarbonate solution.