Automatic preparation method of < 18 > F-labeled polypeptide radiopharmaceutical targeting integrin alpha 6 and kit

Abstract

The invention provides an automatic preparation method of < 18 > F-labeled polypeptide radiopharmaceuticals targeting integrin alpha 6 and a kit. The [< 18 > F] AlF-NOTA-polypeptide is fully automatically produced by adopting a novel independently researched and developed ferrule type radiopharmaceutical synthesis module Mortenon M1 capable of extracting fluorine ions in batches, the preparation method is simple and stable, < 18 > F ions can be extracted in batches according to needs to be added into a reaction, the < 18 > F ions can be produced through an accelerator in one time, and the < 18 > F ions can be repeatedly used. According to the method, efficient automatic production and purification of the medicine targeting the integrin alpha6 are achieved, radioactive exposure is avoided, the cost of single-time synthesis is reduced, economic benefits are improved, and clinical radiopharmaceutical requirements can be met; the probe can specifically recognize tumors with high expression of integrin alpha6, is high in uptake in tumor tissues, and has better targeting property. As a nuclear medicine PET / CT tumor imaging drug, the compound has a good clinical application prospect.

Description

Technical Field

[0001] The present invention relates to the field of radioactive drugs and nuclear medicine technology for tumor imaging, and in particular to a method for targeting tumors with high expression of integrin α6 (such as liver cancer, colorectal cancer, esophageal cancer and nasopharyngeal cancer, etc.) 18 An automated preparation method and kit for F-labeled polypeptide nuclear medicine positron radioactive drugs. Background Art

[0002] Integrins are a class of membrane proteins that mediate intracellular integrin receptors. They are a family of proteins that mediate the adhesion of cells to extracellular matrix proteins. Integrins are expressed on the cell surface and act as regulators of multiple cell functions that are critical to the occurrence, development, and metastasis of solid tumors. Therefore, they are ideal targets for tumor molecular imaging and tumor targeted therapy. Integrins are composed of 18 α subunits and 8 β subunits to form 24 heterodimers. Integrins overexpressed in tumors mainly include αvβ3, αvβ5, α5β1, α4β1, α2β1, α3β1, αvβ6, α6β4, and α6β1. Among them, RGD peptide molecular probes and tumor-targeted drugs targeting integrin αvβ3 are the most widely studied. At present, researchers have successfully developed a variety of RGD peptide radionuclide molecular probes, magnetic resonance molecular probes, ultrasound molecular probes, and optical molecular probes. A considerable number of these RGD peptide molecular probes have entered clinical trials and have achieved good imaging effects in tumor patients.

[0003] Integrin α6 exists in the body as α6β4 and α6β1 dimers. The integrin α6 subunit is overexpressed in a variety of tumors, including breast cancer, liver cancer, lung cancer, colorectal cancer, esophageal cancer, glioma, pancreatic cancer, prostate cancer, head and neck tumors, nasopharyngeal carcinoma, cervical cancer, gastric cancer, renal cancer, pheochromocytoma and paraganglioma. Integrin α6 plays an important role in the migration, invasion and proliferation of tumor cells. At the same time, the GEPIA database shows that the higher the expression level of integrin α6 in tumor patients, the worse the overall survival rate and disease-free survival rate. Therefore, integrin α6 is an important marker for tumor diagnosis and prognosis evaluation. The development of peptide diagnostic reagents targeting integrin α6 is of great significance for the diagnosis of tumors.

[0004] As a non-invasive in vivo imaging technology, nuclear medicine molecular imaging technology can detect abnormalities in the level of diseased genes or proteins before the tumor undergoes anatomical morphological changes, and provide quantitative analysis and real-time evaluation of tumor markers in vivo to guide early diagnosis and personalized precision treatment of the disease. Therefore, it is urgent to develop a peptide radiopharmaceutical targeting integrin α6 with strong targeting specificity and high sensitivity to provide guidance for the early diagnosis and efficacy evaluation of malignant tumors with high expression of integrin α6. Based on this, the present invention studies peptide radiopharmaceuticals targeting integrin α6 [18 F]AlF-NOTA-A6P, [ 18 F]AlF-NOTA-RWY, [ 18 F]AlF-NOTA-RD 2 , and applied it to imaging studies of various tumor-bearing mouse models, showing good prospects for clinical application.

[0005] Nuclide 18 F is used to label radioactive drugs due to its excellent physical properties and the advantage of being available in large quantities from accelerator production. Conventional methods are complicated to operate, take a long time to synthesize, and are difficult to achieve automated synthesis. 18 F notation becomes 18 F is an option for labeling peptide drugs. However, manual labeling has low activity and poses risks to personal radiation safety protection, and the existing modules are produced by accelerators in a single pass. 18 F ions can only be used for one synthesis and consumables 18 OH 2 O is expensive and the automated labeling yield is low, which is difficult to meet clinical needs. In order to improve the efficiency of single synthesis and enhance economic benefits, a radioactive drug targeting integrin α6 that meets the Good Manufacturing Practice (GMP) of pharmaceuticals was developed. 18 F ion card-type radiopharmaceutical synthesis module to achieve single batch production 18 The present invention has developed a simple and high activity preparation method for the purpose of multi-batch synthesis of F ions. 18 The fully automated synthesis method and kit for F-labeled integrin α6-targeting peptide radiopharmaceuticals were developed to meet the needs of clinical research. Summary of the invention

[0006] In view of the shortcomings of the prior art, the present invention aims to provide a new type of 18 The invention discloses an automated preparation method and kit for F-labeled peptide radiopharmaceuticals targeting integrin α6. The method of the invention uses a novel automated synthesis module Mortenon M1 to study [ 18 F]AlF chelate labeling method is used in this module for reaction and purification, and is realized 18 Automated synthesis, large-dose production and clinical application of F-labeled peptide drugs.

[0007] The technical solution adopted by the present invention to solve the technical problem is:

[0008] The first aspect of the present invention is to provide 18 F-labeled polypeptide radiopharmaceutical targeting integrin α6, wherein the polypeptide radiopharmaceutical is any one of the following:

[0009] (1) Peptide radiopharmaceuticals1:18 F]AlF-NOTA-A6P, including polypeptide cyclo[Arg-Trp-Tyr-Asp(NOTA-P EG4-Ethylenediamine)-Dglu-Ala] and bifunctional chelating agent NOTA, as well as fluorine-18 ions and Al atoms, has the following specific structure:

[0010]

[0011] (2) Peptide radiopharmaceuticals 2: 18 F]AlF-NOTA-RWY, including polypeptide Arg-Trp-Tyr-Asp-PEG4-Lys (NOTA) and bifunctional chelating agent NOTA, as well as fluorine-18 ions and Al atoms, has a specific structure of:

[0012]

[0013] (3) Labeled peptide radiopharmaceuticals 3: 18 F]AlF-NOTA-RD 2 , including polypeptide (Arg-Trp-Tyr-Asp-PEG4)2-Ly s-Lys (NOTA) and bifunctional chelating agent NOTA, as well as fluorine-18 ions and Al atoms, the specific structure is:

[0014]

[0015] The radioactive drug of the present invention can specifically recognize tumors and lesions with high expression of integrin α6, has high uptake in tumor tissues, and has a higher tumor / normal tissue ratio (ie, target / non-target ratio).

[0016] The second aspect of the present invention provides a method comprising the above-mentioned 18 A kit for synthesizing a polypeptide radiopharmaceutical targeting integrin α6 labeled with F, the kit comprising: a first reagent bottle containing 10 mL of physiological saline, a second reagent bottle containing 100-150 μg of a precursor, and a second reagent bottle containing 1.0 mL of 2 mM AlCl 3 A third reagent bottle containing HAc, a fourth reagent bottle containing 5 mL of DMSO, a sixth reagent bottle containing 1.5 mL of 70% ethanol by volume, a seventh reagent bottle containing 10 mL of normal saline, 500 mL of sterile water for injection, an anion column, a solid phase extraction column, a pretreatment column solvent or solution, a syringe, a needle, a product receiving bottle and a sterile filter membrane.

[0017] The anion column is specifically a SEP-PAK Light QMA column; the solid phase extraction column is specifically a C18 or HLB column, and the pretreatment column solvent or solution specifically includes ethanol and water for injection.

[0018] The third aspect of the present invention provides the above 18 An automated preparation method for F-labeled polypeptide radiopharmaceuticals targeting integrin α6, the method comprising the following steps:

[0019] (1) The accelerator produces 18 F ion 18 OH 2 O target water is transferred to the transfer bottle;

[0020] (2) Extraction of fluoride ions according to the required amount 18 OH 2 O is transferred to the anion column for capture 18 F ions;

[0021] (3) Elute with saline through a QMA column 18 F ions to the reaction bottle;

[0022] (4) Extraction of precursor / AlCl 3 / HAc / dimethyl sulfoxide (DMSO) mixed solution is added into the reaction bottle, the precursor is a derivative of a polypeptide connected with NOTA, and the NOTA is a triazacyclononane triacetate group;

[0023] (5) heating the reaction bottle to 100°C-110°C for marking;

[0024] (6) diluting with water after cooling, and transferring to a solid phase extraction column for separation and purification;

[0025] (7) washing the solid phase extraction column with injection water to remove organic solvents and impurities;

[0026] (8) The solid phase extraction column is rinsed with 70% ethanol solution and filtered through a sterile filter membrane into a product bottle, and then diluted with physiological saline to obtain the product.

[0027] In a specific embodiment, in step (2), the anion column is a SEP-PAK QMA column.

[0028] In a specific embodiment, in step (4), the precursor is 100-150 μg, 2 mM AlCl 3 The volume of HAc is 5 μl and the volume of DMSO is 3.6 mL.

[0029] In a specific embodiment, in step (6), the solid phase extraction column is a C18 column or an HLB column.

[0030] In a specific embodiment, in step (4), the precursor is cyclo[Arg-Trp-Tyr-Asp(NOTA-PEG4-Ethylenediamine)-Dglu-Ala](NOTA-A6P), and the first reaction liquid is [ 18 F]AlF, the product is [ 18 F]AlF-NOTA-A6P.

[0031] In a specific embodiment, in step (4), the precursor is Arg-Trp-Tyr-Asp-PEG4-Lys(NOTA)(NOTA-RWY), and the first reaction liquid is [ 18 F]AlF, the product is [ 18 F]Al F-NOTA-RWY.

[0032] In a specific embodiment, in step (4), the precursor is (Arg-Trp-Tyr-Asp-PEG4)2-Lys-Lys(NOTA)(NOTA-RD 2 ), the first reaction liquid is [ 18 F]AlF, the product is [ 18 F]AlF-NOTA-RD 2 .

[0033] In a specific embodiment, the preparation method is implemented based on the new domestic cartridge-type automated synthesis module Mortenon M1.

[0034] The method of the present invention can complete the automated radioactive synthesis, and adopts a kit and a novel subpackage 18 The F ion cartridge synthesis module enables simple, high-yield, high-dose automated synthesis 18 F-labeled peptide radiopharmaceuticals can improve economic benefits while providing high-dose products for clinical transformation. A single batch can meet the PET examination needs of more patients.

[0035] The present invention uses a novel automated synthesis module Mortenon M1 to study the use of a card sleeve type [ 18 F]AlF chelation labeling method is automated, and the reaction is obtained by solid phase separation to obtain high-purity 18 F-labeled polypeptide radiopharmaceuticals. The synthesis method of the present invention is applicable to different radioactivity 18 F ions (3.7-74 GBq), solid phase separation [ 18 F]AlF chelate labeling method from transmission 18The total time from the F ion to the preparation and purification of the product is within 30 minutes, the yield after uncorrected decay is as high as 35%-60%, and the radiochemical purity is greater than 99%. This method can achieve simple, high-yield, high-dose automated synthesis. 18 F-labeled peptide radiopharmaceuticals, and the synthesis process using ferrules meets GMP requirements, providing stable and high-quality products for clinical use.

[0036] Beneficial effects of the present invention:

[0037] 1. The targeting of integrin α6 of the present invention 18 F-labeled peptide molecular probes can specifically identify tumors with high expression of integrin α6 (such as liver cancer, colorectal cancer, esophageal cancer and nasopharyngeal cancer, etc.), and use nuclear medicine positron emission tomography (PET) technology to perform real-time dynamic diagnosis and imaging of tumors in a non-invasive, highly sensitive and highly specific manner.

[0038] 2. The automated preparation method of the present invention uses a new type of repackaging product independently developed 18 Mortenon M1, a cartridge-type synthesis module for F ions, enables single-batch production of fluoride ions, multi-batch synthesis, and simple, high-yield, high-dose automated synthesis 18 The synthesis yield of the F-labeled integrin α6 polypeptide radiopharmaceutical is 20-30% higher than the existing automated synthesis yield of aluminum fluoride chelate labeling.

[0039] 3. The present invention provides a kit for a radioactive molecular probe drug targeting integrin α6, which can be conveniently used for automated labeling of novel synthetic modules. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 for 18 F-labeled peptide radiopharmaceuticals [ 18 F] Labeling flow chart of AlF-NOTA-A6P;

[0041] Figure 2 for 18 F-labeled peptide radiopharmaceuticals [ 18 F] Labeling flow chart of AlF-NOTA-RWY;

[0042] Figure 3 for 18 F-labeled peptide radiopharmaceuticals [ 18 F]AlF-NOTA-RD 2 The marking flow chart of

[0043] Figure 4 for 18 Schematic diagram of the automated synthesis module of F-labeled peptide radiopharmaceuticals targeting integrin α6;

[0044] Figure 5 In the embodiment of the present invention, 18 F] High performance liquid chromatography (HPLC) of AlF-NOTA-A6P;

[0045] Figure 6 In the embodiment of the present invention, 18 F]High performance liquid chromatography (HPLC) of AlF-NOTA-RWY;

[0046] Figure 7 In the embodiment of the present invention, 18 F]AlF-NOTA-RD 2 High performance liquid chromatography (HPLC) diagram;

[0047] Figure 8 In the embodiment of the present invention, 18 F] Small animal micro-PET images of the colorectal cancer LS174T tumor-bearing mouse model of AlF-NOTA-A6P;

[0048] Fig. 9 In the embodiment of the present invention, 18 F]AlF-NOTA-A6P biodistribution in nude mice bearing colorectal cancer LS174T tumors (%ID / g; means±SD).

[0049] Fig.10 for 18 Flow chart of the automated synthesis process of F-labeled integrin α6-targeted peptide PET probes; A1, A2, A3, and A4 are connected to spare reagent bottles, A5 is a saline eluent, A6 is a fluoride ion transfer bottle, B1 is a precursor, B2 is 70% ethanol, B3 is a saline solution for dilution, B4 is a product bottle, V1-V10 are three-way valves, R1 is a reaction bottle, S1 and S2 are syringes, and S3 is a micro-syringe with a scale. DETAILED DESCRIPTION

[0050] The technical solution of the present invention is further described in detail below through the accompanying drawings and specific embodiments. The reagents or instruments used without indicating the manufacturer are regarded as conventional products that can be purchased on the market.

[0051] The embodiment of the present invention provides 18 The synthesis method of F-labeled peptide drugs, the process flow diagram is as follows Figure 4 The cyclotron produced 18 F ion 18 OH 2 O target water is transferred to transfer bottle A4, and syringe S3 is used to extract 18 OH2 O target water is transferred to anion QMA column for capture 18 F ions were eluted from the QMA column with 0.3-0.5 mL of saline. 18 F ions were eluted into reaction bottle R1; then mixed solution B1 (100 μg precursor / 30 μl 2 mM AlCl 3 / 5μl HAc / 3.6mL DMSO) is added to the reaction bottle R1; wherein the precursor is a derivative of a polypeptide connected with NOTA; NOTA is triazacyclononane triacetate; labeling synthesis is performed at 100°C, and after cooling, S1 extracts water for injection and adds it to the reaction bottle R1 to dilute the reaction solution, S1 extracts the reaction solution and transfers it to a solid phase extraction column (C18 column in the figure) for solid phase separation and purification, the product is adsorbed on the solid phase extraction column, and the C18 column is cleaned with water for injection extracted by S1 to remove organic solvents and impurities; S1 extracts 70% ethanol in B2 to rinse the solid phase extraction column through a sterile filter membrane into the product bottle B4, and dilutes it with physiological saline to obtain the product injection solution.

[0052] Among them, the anion column can be specifically a SEP-PAK Light QMA column, the precursor-AlCl 3 The molar ratio is 1.45:1, the mass of the precursor is 100 μg, and the solid phase extraction column is specifically a C18 or HLB column.

[0053] In a first specific example, the precursor is cyclo[Arg-Trp-Tyr-Asp(NOTA-PEG4-Ethylenediamine)-Dglu-Ala](NOTA-A6P), and the first reaction liquid is [ 18 F]AlF, the product is [ 18 F]AlF-NOTA-A6P. The schematic diagram of the synthesis process is shown in Figure 1 .

[0054] In a second specific example, the precursor is Arg-Trp-Tyr-Asp-PEG4-Lys(NOTA)(NOTA-RWY), and the first reaction liquid is [ 18 F]AlF, the product is [ 18 F]AlF-NOTA-RWY. The schematic diagram of the synthesis process is shown in Figure 2 .

[0055] In a third specific example, the precursor is (Arg-Trp-Tyr-Asp-PEG4)2-Lys-Lys(NOTA)(NOTA-RD 2 ), the first reaction liquid is [ 18 F]AlF, the product is [ 18 F]AlF-NOTA-RD2 The schematic diagram of the synthesis process is shown in Figure 3 .

[0056] The following is a further detailed description with some specific implementation examples.

[0057] Embodiment 1 18 Automated Synthesis of F]AlF-NOTA-A6P Injection Solution

[0058] The composition of the cartridge reagent: 0.3mL-0.5mL of normal saline (bottle B3), precursor dissolved in DMSO (bottle B1), 1.5mL of 70% ethanol (bottle B2), auxiliary materials including: 500mL of sterile water for injection, anion column (SEP-PAK QMA), solid phase extraction column (C18 column or HLB column), pretreatment column solvent or solution (ethanol and sterile water for injection for activated C18 column or HLB column), syringe, needle, product receiving bottle and sterile filter membrane.

[0059] Assemble the automated synthesis cartridge: rinse the SEP-PAK QMA with 10 mL of water for injection, activate the C18 column with 10 mL of anhydrous ethanol and 10 mL of sterile water for injection, and then assemble bottle B1, bottle B2, bottle B3 and product bottle B4 as well as the SEP-PAK QMA and the activated C18 column on a disposable cartridge (CN 114762728 A).

[0060] Start the automated synthesis device: start the system self-check, and after the self-check, start synthesizing the target product. 18 F ion 18 OH 2 O target water is transferred to transfer bottle A4, and syringe S3 is used to extract 18 OH 2 O target water is transferred to anion QMA column for capture 18 F ions were eluted from the QMA column with 0.3-0.5 mL of saline. 18 F ions were eluted into reaction bottle R1; then mixed solution B1 (100 μg precursor / 30 μl 2 mM AlCl 3 / 5μl HAc / 3.6mL DMSO) was added to the reaction bottle R1; wherein the precursor was a derivative of a polypeptide connected with NOTA; NOTA was triazacyclononane triacetate; the labeling synthesis was performed at 100°C, after cooling, S1 extracted water for injection and added to the reaction bottle R1 to dilute the reaction solution, S1 extracted the reaction solution and transferred it to a solid phase extraction column (C18 column in the figure) for solid phase separation and purification, the product was adsorbed on the solid phase extraction column, and the C18 column was cleaned with water for injection extracted by S1 to remove organic solvents and impurities; S1 extracted 70% ethanol from B2 to rinse the solid phase extraction column through a sterile filter membrane into the product bottle B4, and diluted with physiological saline to obtain [ 18 F]AlF-NOTA-A6 injection. The undecay-corrected yield reached 58.1%±4.1, and the radiochemical purity was higher than 99%.

[0061] Embodiment 2 18 Automated Synthesis of F]AlF-NOTA-RWY Injection

[0062] The composition of the cartridge reagent: 0.3mL-0.5mL of normal saline (bottle B3), precursor dissolved in DMSO (bottle B1), 1.5mL of 70% ethanol (bottle B2), auxiliary materials including: 500mL of sterile water for injection, anion column (SEP-PAK QMA), solid phase extraction column (C18 column or HLB column), pretreatment column solvent or solution (ethanol and sterile water for injection for activated C18 column or HLB column), syringe, needle, product receiving bottle and sterile filter membrane.

[0063] Assemble the automated synthesis cartridge: rinse the SEP-PAK QMA with 10 mL of water for injection, activate the C18 cartridge with 10 mL of anhydrous ethanol and 10 mL of sterile water for injection, and then assemble bottle B1, bottle B2, bottle B3, product bottle B4, SEP-PAK QMA, and the activated C18 cartridge on the disposable cartridge.

[0064] Start the automated synthesis device: start the system self-check, and after the self-check, start synthesizing the target product. 18 F ion 18 OH 2 O target water is transferred to transfer bottle A4, and syringe S3 is used to extract 18 OH 2 O target water is transferred to anion QMA column for capture 18 F ions were eluted from the QMA column with 0.3-0.5 mL of saline. 18 F ions were eluted into reaction bottle R1; then mixed solution B1 (100 μg precursor / 30 μl 2 mM AlCl 3 / 5μl HAc / 3.6mL DMSO) was added to the reaction bottle R1; wherein the precursor was a derivative of a polypeptide connected with NOTA; NOTA was triazacyclononane triacetate; the labeling synthesis was performed at 100°C, after cooling, S1 extracted water for injection and added to the reaction bottle R1 to dilute the reaction solution, S1 extracted the reaction solution and transferred it to a solid phase extraction column (C18 column in the figure) for solid phase separation and purification, the product was adsorbed on the solid phase extraction column, and the C18 column was cleaned with water for injection extracted by S1 to remove organic solvents and impurities; S1 extracted 70% ethanol from B2 to rinse the solid phase extraction column through a sterile filter membrane into the product bottle B4, and diluted with physiological saline to obtain [ 18 F]AlF-NOTA-RWY injection. The undecay-corrected yield reaches 35%-60%, and the radiochemical purity is higher than 98%.

[0065] Example 3 18 F]AlF-NOTA-RD 2 Automated large-dose synthesis of injections

[0066] The composition of the cartridge reagent: 0.3mL-0.5mL of normal saline (bottle B3), precursor dissolved in DMSO (bottle B1), 1.5mL of 70% ethanol (bottle B2), auxiliary materials including: 500mL of sterile water for injection, anion column (SEP-PAK QMA), solid phase extraction column (C18 column or HLB column), pretreatment column solvent or solution (ethanol and sterile water for injection for activated C18 column or HLB column), syringe, needle, product receiving bottle and sterile filter membrane.

[0067] Assemble the automated synthesis cartridge: rinse the SEP-PAK QMA with 10 mL of water for injection, activate the C18 cartridge with 10 mL of anhydrous ethanol and 10 mL of sterile water for injection, and then assemble bottle B1, bottle B2, bottle B3, product bottle B4, SEP-PAK QMA, and the activated C18 cartridge on the disposable cartridge.

[0068] Start the automated synthesis device: start the system self-check, and after the self-check, start synthesizing the target product. 18 F ion 18 OH 2 O target water is transferred to transfer bottle A4, and syringe S3 is used to extract 18 OH 2 O target water is transferred to anion QMA column for capture 18 F ions were eluted from the QMA column with 0.3-0.5 mL of saline. 18 F ions were eluted into reaction bottle R1; then mixed solution B1 (100 μg precursor / 30 μl 2 mM AlCl 3 / 5μl HAc / 3.6mL DMSO) was added to the reaction bottle R1; wherein the precursor was a derivative of a polypeptide connected with NOTA; NOTA was triazacyclononane triacetate; the labeling synthesis was performed at 100°C, after cooling, S1 extracted water for injection and added to the reaction bottle R1 to dilute the reaction solution, S1 extracted the reaction solution and transferred it to a solid phase extraction column (C18 column in the figure) for solid phase separation and purification, the product was adsorbed on the solid phase extraction column, and the C18 column was cleaned with water for injection extracted by S1 to remove organic solvents and impurities; S1 extracted 70% ethanol from B2 to rinse the solid phase extraction column through a sterile filter membrane into the product bottle B4, and diluted with physiological saline to obtain [ 18 F]AlF-NOTA-RD 2 Injection. The undecay-corrected yield is 35%-60% and the radiochemical purity is higher than 98%.

[0069] HPLC method to detect the radiochemical purity of injection: HPLC analysis conditions, analytical column C18 (Autima TM 5μm, 250×4.6mm, Grace, USA), flow rate 1ml / min, gradient elution, mobile phase A: 0.02% v / v trifluoroacetic acid (TFA) in water; mobile phase B: 0.04% v / v trifluoroacetic acid (TFA) in acetonitrile; 0–3min mobile phase A / mobile phase B=95:5; 3.01-20min mobile phase A / mobile phase B=60:40; 20.01-30min mobile phase A / mobile phase B: 95:5.

[0070] Example 4 PET imaging of colorectal cancer tumor-bearing mice

[0071] Colorectal cancer LS174T tumor-bearing mice with moderate tumor size were injected with 3.7 MBq [ 18 After the injection of F]AlF-NOTA-A6P, microPET static scanning and image reconstruction were performed at 30, 60 and 120 minutes. The regions of interest (ROIS) such as tumor, muscle, heart, and liver were outlined, and the percentage of tissue dose per gram (%ID / g) was calculated. The resulting PET images are shown in the figure below. Figure 8 As shown, injection [ 18 Tumor uptake can be observed 30 minutes after administration of F]AlF-NOTA-A6P, and the tumor uptake reaches the highest value at 60 minutes, indicating that the molecular probe has good targeting and a good retention time in the tumor. The uptake in organs such as the kidney and bladder is relatively high, indicating that the drug is metabolized by the kidneys, cleared from the blood pool quickly, and has low liver uptake, and has excellent in vivo pharmacokinetic properties.

[0072] [ 18 F]AlF-NOTA-RWY and [ 18F]AlF-NOTA-RD 2 Similar features were also shown in PET imaging of colorectal cancer LS174T tumor-bearing mice.

[0073] The protection content of the present invention is not limited to the above embodiments. Without departing from the spirit and scope of the inventive concept, changes and advantages that can be thought of by those skilled in the art are included in the present invention and are protected by the attached claims.

Claims

1. A 18 F-labeled polypeptide radiopharmaceutical targeting integrin α6, wherein the polypeptide radiopharmaceutical is any one of the following: (1) Peptide radiopharmaceuticals1: 18 F]AlF-NOTA-A6P, including polypeptide cyclo[Arg-Trp-Tyr-Asp(NOTA-PEG4-Ethylenediamine)-Dglu-Ala] and bifunctional chelating agent NOTA, as well as fluorine-18 ions and Al atoms, has a specific structure as follows: (2) Peptide radiopharmaceuticals 2: 18 F]AlF-NOTA-RWY, including the peptide Arg-Trp-Tyr-Asp-PEG4- Lys(NOTA) and bifunctional chelating agent NOTA, as well as fluorine-18 ions and Al atoms, the specific structure is: (3) Peptide radiopharmaceuticals 3: 18 F]AlF-NOTA-RD2, including polypeptide (Arg-Trp-Tyr-Asp-PEG4)2-Lys-Lys(NOTA) and bifunctional chelating agent NOTA, as well as fluorine-18 ions and Al atoms, has the following specific structure:

2. A 18 The method for automatically preparing F-labeled polypeptide radiopharmaceuticals targeting integrin α6 is characterized in that: The method comprises: (1) The accelerator produces 18 F ion 18 O-H2O target water is transferred to the transfer bottle; (2) Extraction of fluoride ions according to the required amount 18 O-H2O is transferred to the anion column and captured 18 F ions; (3) Elute with saline through a QMA column 18 F ions to the reaction bottle; (4) extracting a precursor / AlCl3 / HAc / dimethyl sulfoxide mixed solution and adding it to a reaction bottle, wherein the precursor is a derivative of a polypeptide connected with NOTA, wherein NOTA is a triazacyclononane triacetate; (5) heating the reaction bottle to 100°C-110°C for marking; (6) diluting with water after cooling, and transferring to a solid phase extraction column for separation and purification; (7) washing the solid phase extraction column with injection water to remove organic solvents and impurities; (8) The solid phase extraction column is rinsed with 70% ethanol solution and filtered through a sterile filter membrane into a product bottle, and then diluted with physiological saline to obtain the product.

3. According to claim 2 18 The method for automatically preparing F-labeled polypeptide radiopharmaceuticals targeting integrin α6 is characterized in that: In the step (2), the anion column is a SEP-PAK QMA column.

4. according to claim 2 18 The method for automatically preparing F-labeled polypeptide radiopharmaceuticals targeting integrin α6 is characterized in that: In the step (4), the precursor is 100-150 μg, 2 mM AlCl3 is 20-30 μl, the volume of HAc is 5 μl, and the volume of DMSO is 3.6 mL.

5. According to claim 2 18 The method for automatically preparing F-labeled polypeptide radiopharmaceuticals targeting integrin α6 is characterized in that: In the step (6), the solid phase extraction column is a C18 column or an HLB column.

6. According to claim 2 18 The method for automatically preparing F-labeled polypeptide radiopharmaceuticals targeting integrin α6 is characterized in that: In the step (4), the precursor is cyclo[Arg-Trp-Tyr-Asp(NOTA-PEG4- Ethylenediamine)-Dglu-Ala], the first reaction liquid is [ 18 F]AlF, the product is [ 18 F]AlF-NOTA- A6P.

7. According to claim 2 18 The method for automatically preparing F-labeled polypeptide radiopharmaceuticals targeting integrin α6 is characterized in that: In the step (4), the precursor is Arg-Trp-Tyr-Asp-PEG4-Lys (NOTA), and the first reaction liquid is [ 18 F]AlF, the product is [ 18 F]Al F-NOTA-RWY.

8. According to claim 2 18 The method for automatically preparing F-labeled polypeptide radiopharmaceuticals targeting integrin α6 is characterized in that: In the step (4), the precursor is (Arg-Trp-Tyr-Asp-PEG4)2-Lys-Lys(NOTA), and the first reaction liquid is [ 18 F]AlF, the product is [ 18 F]AlF-NOTA-RD2.

9. According to any one of claims 2 to 8 18 The method for automatically preparing F-labeled polypeptide radiopharmaceuticals targeting integrin α6 is characterized in that: The preparation method is based on a new domestically produced cartridge-type automated synthesis module, Mortenon M1.

10. A method for use in the method according to claim 1 18 A kit for synthesizing F-labeled polypeptide radiopharmaceuticals targeting integrin α6, characterized in that: The kit includes: physiological saline, precursor, AlCl3, HAc, DMSO and 70% ethanol; Preferably, the kit further comprises an anion column, a solid phase extraction column, solvents and solutions for pre-treating the column, a syringe, a needle, a product bottle and a sterile filter membrane; More preferably, the anion column is a SEP-PAK QMA column, the solid phase extraction column is a C18 column or an HLB column, and the solvent or solution of the pretreatment column includes ethanol and water for injection.

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