Preparation method, kit and application of Al18F tracer integrated freeze-dried powder

Abstract

The invention provides a preparation method of Al18F tracer integrated freeze-dried powder, a kit and application thereof, and the preparation method comprises the following steps: dissolving 1-10 mg of a precursor by using deionized water, adding a pH buffer solution with the concentration of 0.2-0.5 M and an AlCl3 solution with the concentration of 10 mM, with the ratio of the amount of substance of AlCl3 to the amount of substance of the precursor being 0.8-1.0, then continuously adding a stabilizer solution with the concentration of 0.5 M and 1-4 mg of an excipient, and uniformly stirring to obtain the Al18F tracer integrated freeze-dried powder. And finally, adding deionized water to enable the concentration of the precursor to be 0.5-1.0 mM, uniformly mixing the solution, sub-packaging the solution into 1.5 mL cryopreservation tubes or 2 mL penicillin bottles according to a certain volume, and then putting the cryopreservation tubes or the penicillin bottles into a vacuum freeze dryer to remove water at a low temperature to obtain powder or freeze-dried cakes, and the container is a freeze-dried powder / cake storage container and is used for storing the freeze-dried powder / cakes. Meanwhile, the reaction container is also used during radioactive labeling.

Description

Technical Field

[0001] The present invention relates to the technical field of preparation of PET tracer products, and in particular to an Al 18 Preparation method, kit and application of F tracer integrated freeze-dried powder. Background Art

[0002] Positron emission tomography based on active small molecules (such as peptides) is increasingly used in the functional diagnosis of diseases, especially in tumors, highlighting its huge potential for clinical transformation. 68 Ga labeling 18 F has unique advantages, such as long half-life, convenient central distribution, and long-distance transportation; high yield, usually in the hundreds of millicuries to several curies; high image quality, high spatial resolution, and easy to find tiny lesions. 18 The F marking method is basically based on 18 The F-labeled synthon is then coupled with a biologically active small molecule. The disadvantages are that it is time-consuming and labor-intensive, with low yields, which is not conducive to clinical transformation. In addition, due to structural changes, the activity of the biologically active molecule changes, that is, the lipid-water partition coefficient changes, which ultimately leads to huge changes in the tracer's targeting and biodistribution.

[0003] Al 18 The F mark is first passed 18 F anion and Al 3+ Ion complexation to form AlF 2+ ions, and then use NOTA-ligand to chelate them, finally forming AlF-NOTA-ligand, achieving the purpose of NOTA-ligand 18 F mark, which corresponds to 68 Ga-NOTA-ligands have a high degree of similarity in terms of biological activity. Currently, manual synthesis has the following disadvantages: high occupational exposure of operators, poor reproducibility, production methods that cannot meet GMP requirements, and low output that cannot meet clinical needs. At the same time, the existing modular flow path architecture is not suitable for Al 18 The results of automated synthesis of F tracer are: low labeling rate, low specific activity, etc.; and the disposable cartridge reagents are expensive, which is not conducive to the research and development of tracers.

[0004] In addition, due to Al 18 F process requirements, in the pre-synthesis preparation stage, usually have to prepare a variety of reagents such as pH buffer solution, AlCl3 stock solution, stabilizer and organic solvent, and then add them manually in sequence. The production process is cumbersome and prone to errors, resulting in the failure of tracer production. Secondly, there is currently no universal process formula for Al 18 F labeling method, usually corresponding to a specific Al18 F tracers have specific actual formulations and process requirements and cannot achieve Al 18 General reagent formula and standardized preparation process for F labeling methodology.

[0005] Patent CN102295685B discloses a 18 F-labeled PRGD2 compound, its kit, kit preparation method and application, relates to a PET imaging agent, preparation method and application, and belongs to the technical field of radiopharmaceuticals and nuclear medicine. The kit includes NOTA-PRGD2 and aluminum chloride, the molar ratio of NOTA-PRGD2 to aluminum chloride is (1-10):1, and it is in the form of freeze-dried powder. Preliminary studies have shown that the kit is simple to label, easy to operate, low in cost, short in time, and can make the prepared complex, 18 F-FAl-NOTA-PRGD2, with 18 F-FPPRGD2 has the same biological properties and is expected to be promoted and applied clinically. However, the following problems still exist: the freeze-dried kit contains only precursors and aluminum, but no buffer salts. In the subsequent labeling process, additional pH buffer solutions and organic solvents must be added, which is cumbersome to operate; the freeze-dried kit does not contain stabilizers (such as Vc or gentisic acid), and radioactive self-decomposition byproducts may occur at high doses; and the amount of effective substances in the freeze-dried kit in the patent embodiment is about 10-20ug. Without the support of freeze-dried powder excipients, the freeze-dried form and stability of the reagents in the freeze-dried kit are difficult to guarantee.

[0006] Based on the above problems, the present invention proposes an Al 18 Preparation method, kit and application of F tracer integrated freeze-dried powder. Summary of the invention

[0007] The object of the present invention is to provide an Al 18 The preparation method, kit and application of F tracer integrated freeze-dried powder use the masking effect of dicarboxylic acid in the labeling process to make Al 18 Compared with the traditional labeling process, the F labeling rate is significantly improved, reaching 65%-75%. The integrated freeze-dried powder / cake used in the present invention contains 18 All the ingredients required for the F labeling process are in an EP tube or vial, and the container can be used as a container for the lyophilized powder and a reaction container at the same time, which not only greatly simplifies the cumbersome reagent filling process in the preparation process of radioactive labeling, but also reduces the material loss in the process of transferring the precursor from the dissolution in the container to the reaction tube compared to the conventional operation process.

[0008] In a first aspect, the present invention provides an Al 18The preparation method of F tracer integrated freeze-dried powder comprises the following steps: dissolving 1-10 mg of a precursor by using deionized water or an ethanol aqueous solution with a volume concentration of 10-20%, adding 10-25 uL of a pH buffer solution with a concentration of 0.2-0.5 M and 5-12 uL of an AlCl3 solution with a concentration of 10 mM, wherein the molar ratio of aluminum to the precursor is 0.8-1.0, then continuing to add 10-25 uL of a stabilizer solution with a concentration of 0.5 M and 1-4 mg of an excipient, and finally adding deionized water to make the concentration of the precursor 0.5-1.0 mM, mixing the above solutions evenly and then packaging them, and then placing them in a freeze vacuum dryer to remove water at low temperature into a freeze-dried cake;

[0009] Wherein, the structural formula of the precursor is shown in Formula 1:

[0010] The molecular weight of the precursor is an Affibody, a polypeptide, a polypeptide-like structure or a small molecule structure not exceeding 10 KDa.

[0011] The ethanol water solvent used is mainly used to provide solubility aid for the precursor with poor water solubility.

[0012] Furthermore, the precursor includes any one of the polypeptides NOTA-DUPA-Pep, NOTA-FAPI-04, NOTA-FAPI-46, NOTA-FAPI-74, NOTA-FAPI-2286, NOTA-Octreotide, NOTA-TATE, NOTA-TOC, NOTA-JR11, NOTA-LM3, NOTA-AlfatideII, NOTA-Pentixafor, NOTA-DK222, NOTA-WL12, NOTA-PCP1, and NOTA-PCP2.

[0013] Furthermore, the pH of the pH buffer solution is 4.0-4.2, the concentration is 0.2-0.5M, and the volume dispensed into each container is 20-25uL; the corresponding solute in the buffer solution is a non-volatile solute.

[0014] Furthermore, the solute in the pH buffer solution is a non-volatile dicarboxylate or aminosulfonate, and the solute includes any one of succinic acid, glutaric acid, potassium hydrogen phthalate (KHP), ascorbic acid (Vc), 4-hydroxyethylpiperazineethanesulfonic acid (HEPPS) and 2-morpholineethanesulfonic acid (MES).

[0015] Furthermore, the solvent system in the AlCl3 solution is a 0.1M acetic acid-sodium acetate pH buffer system with a pH of 4.0; and the molar ratio of the amount of AlCl3 to the amount of the precursor in the mixed solution is 0.8-1.0.

[0016] Furthermore, the stabilizer solution is an ascorbic acid aqueous solution or a gentisic acid aqueous solution with a concentration of 0.5M; and the volume of the stabilizer solution dispensed into each container is 5-10uL.

[0017] Furthermore, the excipient is any one of trehalose, mannitol and glucose; and the mass of the excipient packed into each container is 1-4 mg.

[0018] In another aspect, the present invention provides an Al 18 F tracer integrated lyophilized powder kit, the Al 18 The integrated freeze-dried powder of F tracer is prepared by the aforementioned method. The preparation method of the kit comprises the following steps: 18 The integrated lyophilized powder of F tracer is packaged in 2 mL vials or encapsulated in 1.5 mL cryotubes, and then placed in a freeze vacuum dryer to remove water at low temperature into a lyophilized cake, which is packaged into a lyophilized medicine box and stored at low temperature. The final state is a white fluffy, integrated lyophilized cake or white fluffy powder; the vial or cryotube is not only a storage container for the lyophilized powder / cake, but also a reaction container for radioactive labeling.

[0019] Furthermore, the volume of the precursor solution dispensed into each tube or bottle is 100-200 uL.

[0020] On the other hand, the present invention also provides the Al 18 F tracer all-in-one freeze-dried powder and Al 18 Application of the F tracer integrated freeze-dried powder kit in the preparation of PET imaging agents.

[0021] The beneficial effects of the present invention are:

[0022] The present invention utilizes a non-volatile dibasic organic acid such as a succinic acid buffer system, which can not only realize the freeze-drying of the freeze-dried medicine kit buffer salt, but also, compared with the acetate buffer system, the glacial acetic acid component will volatilize during the freeze-drying process. The pH of the medicine kit system does not change before and after the freeze-drying of the succinic acid system buffer, which is convenient for the integrated freeze-drying of the buffer reagent; secondly, the succinic acid dibasic organic acid system can play a certain role as a masking agent, which can mask the excess free Al 3+ ions, preventing excessive free Al 3+ Ions competitively react with NOTA-ligands, thereby increasing Al 18 F 2+ The ions react more efficiently with the NOTA-ligand to improve the labeling efficiency; in addition, stabilizers such as ascorbic acid or gentisic acid are added to the freeze-dried kit to prevent radioactive self-decomposition, which can eliminate Al 18F removes oxygen free radicals in the reaction system during the labeling process; at the same time, the addition of freeze-dried powder excipients ensures the state and stability of the contents of the freeze-dried tube / bottle without affecting the labeling rate.

[0023] The succinic acid buffer system used in the present invention is safer than the phthalic acid buffer system in the prior art. Phthalic acid may be a chemical manifestation related to plasticizers, and according to the product purification process of patent CN118045208A, it is known that the substance cannot be completely removed, and the product injection may contain the substance. The succinic acid used in the present invention is an endogenous substance in the human body, and the purification process of the present invention can also completely remove the substance, so the injection produced by the process of the present invention has better safety.

[0024] Use the kit provided by the present invention to carry out Al 18 F tracer labeling, the operation steps are simple, just add a certain volume of organic solvent to the sealed cryopreservation tube or vial to dissolve the lyophilized powder, and then add 18 F target water or 18 F concentrate is sufficient, and no additional reagents need to be added. Whether it is automatic addition or manual addition, the tedious process of adding reagent solutions is eliminated, which greatly reduces the Al caused by human errors in the filling process. 18 F failed to mark and in the broader 18 The F activity range (such as tens of millicuries to 2.5 curies) remains relatively stable and has a high labeling efficiency (corrected radiochemical labeling rate is about 70-80%, and the yield ranges from tens to 1500 millicuries, depending on the dosage). 18 F target water activity), radiochemical purity> 95%. According to the literature reports and patents known so far, the technical effect (yield and output) of this invention is significant (currently usually Al 18 The F labeling rate is 30-40%, and the output is usually in the range of tens to hundreds of millicuries. The implementation of the invention can provide possibilities and technical support for large-scale production and centralized distribution of radiopharmaceutical manufacturers.

[0025] The present invention has been optimized to achieve high-efficiency labeling based on a small amount of precursor, thereby achieving low radiation dose. 18 The preparation of high specific activity tracers under F ion starting conditions is mainly used in preclinical small animal imaging studies, that is, to produce as much final tracer as possible with the least amount of precursor. 18F starting dose, to produce as much tracer as possible, thereby increasing the specific activity, but this method is an extensive production method, producing a large number of tracers (usually hundreds of millicuries), only a few millicuries are needed, causing waste and creating an additional radiation protection burden.

[0026] By using the kit and the corresponding preparation method of the present invention, the radioactive activity directly contacted by the operator is reduced, and the radiation damage caused by unnecessary overexposure of the operator is avoided. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings:

[0028] Figure 1 For the present invention A1 18 Flow chart of the preparation method of F-PSMA-BCH;

[0029] Figure 2 The state of the NOTA-PSMA-BCH lyophilized powder in the kit of Example 1 of the present invention in a 1.5 mL cryotube;

[0030] Figure 3 The state of the NOTA-PSMA-BCH lyophilized powder in the 2 mL vial in the kit of Example 1 of the present invention;

[0031] Figure 4 The Al was prepared from the kit of Example 1 of the present invention. 18 Radio-TLC18F ion residual test results of F-PSMA-BCH injection;

[0032] Figure 5 The Al was prepared from the kit of Example 1 of the present invention. 18 Radiochemical purity of F-PSMA-BCH injection by radio-HPLC quality control results;

[0033] Figure 6 Prostate cancer patients receiving Al 18 PET imaging results of the affected area after F-PSMA-BCH injection. DETAILED DESCRIPTION

[0034] The technical solution of the present invention is described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0035] The present invention provides an Al 18 The preparation method of F tracer integrated freeze-dried powder comprises the following steps: dissolving 1-10 mg of a precursor by using deionized water or an ethanol aqueous solution with a volume concentration of 10-20%, adding 10-25 uL of a pH buffer solution with a concentration of 0.2-0.5 M and 5-12 uL of an AlCl3 solution with a concentration of 10 mM, wherein the molar ratio of aluminum to the precursor is 0.8-1.0, then continuing to add 10-25 uL of a stabilizer solution with a concentration of 0.5 M and 1-4 mg of an excipient, and finally adding deionized water to make the concentration of the precursor 0.5-1.0 mM, mixing the above solutions evenly and then packaging them, and then placing them in a freeze vacuum dryer to remove water at low temperature into a freeze-dried cake;

[0036] Wherein, the structural formula of the precursor is shown in Formula 1:

[0037] The molecular weight of the precursor is an Affibody, a polypeptide, a polypeptide-like structure or a small molecule structure not exceeding 10 KDa.

[0038] Among them, the precursor includes any one of Affibody class NOTA-HER2-BCH, peptide class NOTA-PSMA-BCH, NOTA-DUPA-Pep, NOTA-FAPI-04, NOTA-FAPI-46, NOTA-FAPI-74, NOTA-FAPI-2286, NOTA-Octreotide, NOTA-TATE, NOTA-TOC, NOTA-JR11, NOTA-LM3, NOTA-AlfatideII, NOTA-Pentixafor, NOTA-DK222, NOTA-WL12, NOTA-PCP1, and NOTA-PCP2.

[0039] More specifically, NOTA-PSMA-BCH is mostly used for prostate tumor diagnosis, and its structural formula is as follows:

[0040]

[0041] NOTA-DUPA-Pep is mostly used for the diagnosis of prostate cancer. Its structural formula is as follows:

[0042]

[0043] NOTA-FAPI-04 targets fibroblasts in the tumor microenvironment and is mostly used for the diagnosis of digestive system tumors. The structural formula is as follows:

[0044]

[0045] NOTA-FAPI-46 targets fibroblasts in the tumor microenvironment and is mostly used for the diagnosis of digestive system tumors. The structural formula is as follows:

[0046]

[0047] NOTA-FAPI-74 targets fibroblasts in the tumor microenvironment and is mostly used for the diagnosis of digestive system tumors. The structural formula is as follows:

[0048]

[0049] NOTA-FAPI-2286 targets fibroblasts in the tumor microenvironment and is mostly used for the diagnosis of digestive system tumors. The structural formula is as follows:

[0050]

[0051] NOTA-Octreotide targets tumor somatostatin receptors and is mostly used in the diagnosis of neuroendocrine tumors. The structural formula is as follows:

[0052]

[0053] NOTA-TATE targets tumor somatostatin receptors and is mostly used in the diagnosis of neuroendocrine tumors. The structural formula is as follows:

[0054]

[0055] NOTA-TOC targets tumor somatostatin receptors and is mostly used in the diagnosis of neuroendocrine tumors. The structural formula is as follows:

[0056]

[0057] NOTA-JR11 targets tumor somatostatin receptors and is mostly used in the diagnosis of neuroendocrine tumors. The structural formula is as follows:

[0058]

[0059] NOTA-LM3 targets tumor somatostatin receptors and is mostly used for the diagnosis of neuroendocrine tumors. It is mostly used for the diagnosis of neuroendocrine tumors. The structural formula is as follows:

[0060]

[0061] NOTA-Alfatide II targets the αvβ3 receptor of tumor neovascularization and is mostly used in the diagnosis of non-small cell lung cancer. The structural formula is as follows:

[0062]

[0063] NOTA-Pentixafor targets the tumor chemokine CXCR4 receptor and has the following structural formula:

[0064]

[0065] NOTA-DK222 targets the tumor immune microenvironment PD-L1 receptor imaging, and its structural formula is as follows:

[0066]

[0067] NOTA-WL12 targets the tumor immune microenvironment PD-L1 receptor imaging, and its structural formula is as follows:

[0068]

[0069] NOTA-PCP1 targets the tumor immune microenvironment PD-L1 receptor imaging, and its structural formula is as follows:

[0070]

[0071] NOTA-PCP2 targets the tumor immune microenvironment PD-L1 receptor imaging, and its structural formula is as follows:

[0072]

[0073] The pH of the pH buffer solution is 4.0-4.2, the concentration is 0.2-0.5M, and the volume of each cryotube or vial is 20-25uL; the corresponding solute in the buffer solution is a non-volatile solute; the solute in the pH buffer solution is a non-volatile dicarboxylate or aminosulfonate, and the solute includes any one of succinic acid, glutaric acid, potassium hydrogen phthalate (KHP), ascorbic acid (Vc), 4-hydroxyethylpiperazineethanesulfonic acid (HEPPS) and 2-morpholineethanesulfonic acid (MES); The solvent system in the AlCl3 solution is a 0.1M acetic acid-sodium acetate pH buffer system with a pH of 4.0; and the molar ratio of the amount of AlCl3 to the precursor in the mixed solution is 0.8-1.0; the stabilizer solution is an ascorbic acid aqueous solution or a gentisic acid aqueous solution with a concentration of 0.5M; and the volume allocated to each cryogenic tube or vial is 5-10uL; the excipient is any one of trehalose, mannitol, and glucose; and the mass of the excipient allocated to each cryogenic tube or vial is 1-4mg.

[0074] The present invention also provides an Al 18 F tracer integrated lyophilized powder kit, the Al 18 The integrated freeze-dried powder of F tracer is prepared by the aforementioned method. The preparation method of the kit comprises the following steps: 18The integrated lyophilized powder of F tracer is packaged in 2mL vials or encapsulated in 1.5mL cryotubes, and then placed in a freeze vacuum dryer to remove water at low temperature into a lyophilized cake, which is packaged into a lyophilized medicine box and stored at low temperature. The final state is a white fluffy, integrated lyophilized cake or white fluffy powder; the vial or cryotube is not only a storage container for the lyophilized powder / cake, but also a reaction container for radioactive labeling; the volume of the solution of the precursor packaged in each tube or bottle is 100-200uL.

[0075] The present invention will be further explained below using specific embodiments:

[0076] Example 1

[0077] This embodiment provides a method for preparing Al 18 Preparation method of integrated lyophilized powder of F-PSMA-BCH tracer, combination of contents packaged in kit thereof, and Al 18 F-PSMA-BCH radiolabeling process, (process as Figure 1 shown) include:

[0078] 1. Preparation steps of integrated freeze-dried powder:

[0079] First, 5 mg of the precursor NOTA-PSMA-BCH was dissolved in 1 mL of deionized water, followed by the addition of 1 mL of 0.5 M succinic acid buffer solution at pH 4.2, 900 uL of 10 mM AlCl3 solution, 250 uL of 0.5 M ascorbic acid stabilizer solution, and 200 mg of mannitol excipient. Finally, deionized water (7.85 mL) was added to make the precursor concentration in the mixed solution 0.5 mM, and then the mixture was fully mixed. The above solution was dispensed into 1.5 mL cryovials (30 tubes) and 3 mL vials (20 bottles) at a volume of 200 uL per tube, and placed in a freeze vacuum dryer to remove water at low temperature to obtain a freeze-dried cake ( Figure 2 and Figure 3 );

[0080] 2. The contents of the kit package:

[0081] Lyophilized powder tube A (1.5mL (with threaded cap) cryotube), physiological saline bag B (Shandong Hualu, 100mL), injection water bottle C (Shandong Hualu, 500mL), ethanol solution bottle D (Aladdin, 50mL), N,N-dimethylformamide reagent bottle E (Aladdin, 2mL), product bottle F (Jiangsu Huayi, 25mL), QMA column N (Jiangsu Huayi, PS-HCO3), SPE product purification column M (Waters, Sep-pakLight) and sterile filter membrane Y (Sartorius, 25mm, S7575), disposable syringe (Shandong Weigao, 10mL), exhaust needle (Zhejiang Kangdelai, 20G, 1.2*38mm).

[0082] 3.Al 18 F radiolabeling is achieved by the following automated process control:

[0083] (1) After the accelerator bombardment is completed, the target water is transferred to the target water receiving bottle and passed through the QMA column. 18 The F ions were adsorbed and blown dry, then rinsed with 120uL of normal saline and concentrated into a 1.5mL integrated freeze-drying tube.

[0084] (2) Then, 300 μL of DMF solvent was added to the reaction system, and the mixture was bubbled and mixed evenly.

[0085] (3) Heat the sealed tube to 100°C for 7 min. After heating, cool to below 50°C.

[0086] (4) Transfer the reaction solution in the reaction tube to the transfer tube, and then use 7 mL of deionized water to wash the reaction tube and dilute the reaction system;

[0087] (5) Purify the product using a solid phase extraction column (model Waters C18 Light);

[0088] (6) washing the solid phase extraction column with 1 mL of 75% ethanol aqueous solution to desorb the product adsorbed on the column and collect it in a 25 mL vial. Finally, diluting it with 9 mL of normal saline, filtering it with an online sterile filter membrane during the product washing process, and preparing a sterile injection solution for injection;

[0089] (7) Final measurement of Al 18 Radioactivity of F-PSMA-BCH injection and starting amount 18 F radioactivity, and thus calculate the radiolabeling rate. The calculation formula is:

[0090] Radioactive labeling rate (%) = total activity of product solution (mCi) / 18 F initial activity (mCi)*100%;

[0091] The Al prepared by the above process18 F-PSMA-BCH kit, 3 batches of Al prepared consecutively 18 The radiolabeling rate measured by F-PSMA-BCH tracer was between 65% and 75%, and the fluoride ion content measured by radio-TLC was no more than 3% ( Figure 4 ), radiochemical purity determined by Radio-HPLC>95% ( Figure 5 ), and the quality control parameters are shown in Table 1.

[0092] Table 1

[0093]

[0094]

[0095] Example 2

[0096] When the precursor is NOTA-PSMA-BCH, the effect of the amount of the precursor used on the radioactive labeling rate is studied. Based on Example 1, the amount of the precursor used is adjusted. The steps are as follows:

[0097] (1) When the precursor in Example 1 is NOTA-PSMA-BCH, the amount of the precursor is adjusted to 1.5, 4, 5, 7.5, and 10 mg, respectively, and the amount of the AlCl3 solution is also adjusted accordingly. The other preparation processes remain unchanged, and freeze-dried powders with corresponding NOTA-PSMA-BCH contents of 30, 80, 100, 150, and 200 ug are prepared, respectively.

[0098] (2) The corresponding reagent kit packaging contents are the same as those in Example 1.

[0099] (3)Al 18 The steps of F radiolabeling were the same as those in Example 1.

[0100] (4) Five batches were prepared using a freeze-dried powder kit containing different amounts of precursors. The radiolabeling rates are shown in Table 2.

[0101] Table 2

[0102]

[0103]

[0104] Combined with the above table, the labeling rate increases with the increase of the amount of precursor; when the amount of precursor is greater than 100ug, the labeling rate remains basically stable at about 75%. In summary, as a preferred embodiment, the content of the lyophilized powder precursor is 100ug, at which time a high radioactive labeling rate can be maintained.

[0105] Example 3

[0106] When the precursor is NOTA-PSMA-BCH, the effect of the type and amount of excipients on the radiolabeling rate is studied. Based on Example 1, the type and amount of excipients are adjusted. The steps are as follows:

[0107] (1) The masses of the excipients in Example 1 were adjusted to 1, 2, and 4 mg, respectively, and the types of the excipients were adjusted to trehalose and mannitol. The other preparation processes remained unchanged, and freeze-dried powders with corresponding excipient contents of 1, 2, and 4 mg were prepared, respectively.

[0108] (2) The corresponding reagent kit packaging contents are the same as those in Example 1.

[0109] (3)Al 18 The steps of F radiolabeling were the same as those in Example 1.

[0110] (4) Using a freeze-dried powder kit containing different excipient types and doses, seven batches were prepared continuously. The radiolabeling rates are shown in Table 3.

[0111] Table 3

[0112]

[0113]

[0114] Conclusion: The dosage of the two excipients between 1-4 mg has no significant effect on the labeling rate. In summary, the 4 mg content is preferred.

[0115] Example 4

[0116] When the precursor is NOTA-PSMA-BCH, the effect of the type and amount of organic solvent on the radioactive labeling rate is studied. Based on Example 1, the type and amount of organic solvent are adjusted. The steps are as follows:

[0117] (1) The freeze-dried powder preparation process is the same as that in Example 1.

[0118] (2) The corresponding reagent kit packaging contents are the same as those in Example 1.

[0119] (3)Al 18 During the radioactive labeling process, the volume of the organic solvent and the additional water added to the cryopreservation tube was adjusted by changing the control program (see Table 4, the dosage column and the additional water column for details), and the remaining operating steps were the same as those in Example 1.

[0120] (4) By adjusting the type and ratio of the organic solvent in the radiolabeling reaction system, 12 batches were prepared continuously. The radiolabeling rates are shown in Table 4.

[0121] Table 4

[0122]

[0123]

[0124] Combined with the above table, increasing the content of organic solvent in the reaction system can significantly improve the labeling rate; the above-mentioned organic solvents have no obvious difference in labeling rate when the content is the same; in summary, as a preferred option, DMSO or DMF is selected as the organic solvent added in the labeling process. Although the labeling rates of acetonitrile and ethanol are similar, due to the relatively low boiling points of these two solvents, during the sealing and heating process, compared with DMSO and DMF, sealing and heating failure is more likely to occur, which ultimately causes labeling failure.

[0125] Example 5

[0126] When the precursor is NOTA-PSMA-BCH, the effect of the type of buffer solution and pH value on the radioactive labeling rate is studied. Based on Example 1, the type of buffer solution is adjusted. See Table 5 below for details:

[0127] (1) The type of buffer solution in Example 1 and the pH of the buffer solution when preparing the mixed solution were adjusted to be as shown in Table 5, and the other preparation processes remained unchanged, and freeze-dried powders containing different buffer systems were prepared respectively.

[0128] (2) The corresponding reagent kit packaging contents are the same as those in Example 1.

[0129] (3)Al 18 The steps of F radiolabeling were the same as those in Example 1.

[0130] (4) Lyophilized powder was prepared by adjusting the type of buffer solution and the pH of the buffer solution when preparing the mixed solution. Eleven batches were prepared continuously. The radiolabeling rate is shown in Table 5.

[0131] Table 5

[0132]

[0133] Combined with the above table, the three pH buffer systems of tartaric acid, citric acid, and malonic acid may be due to the buffer salt and Al 3+ Significant chelation occurs, resulting in a low radiolabeling rate (<5%), so it is not suitable as a pH buffer for Al 18F labeling; pH buffer solution systems around pH 4.0 (succinic acid, Vc system, KHP system) all showed high labeling rates, but with the drastic change in pH, the labeling rates of the Vc and KHP systems dropped sharply, but the succinic acid system showed a relatively wide pH tolerance. Although the pH of the acetic acid buffer system is 4.0 when preparing the mixed solution, the buffer system contains volatile acetic acid components. During the freeze-drying process, the acetic acid evaporates, causing the pH value to change drastically after the freeze-dried powder is re-dissolved, resulting in a low labeling rate (<5%). In summary, the 0.5M succinic acid pH buffer solution system is preferred as Al 18 F-labeled buffer system (although the labeling rate of KHP pH 4.0 is basically the same as that of succinic acid, it is extremely difficult to completely remove KHP during the purification of subsequent product injections. This substance is a non-endogenous substance related to plasticizers and has potential biosafety issues).

[0134] Example 6

[0135] When the precursor is NOTA-PSMA-BCH, the effect of the volume and material of the reaction tube on the radiolabeling rate is studied. Based on Example 1, the volume of the reaction tube is adjusted. See Table 6 below for details:

[0136] (1) The container used for the final packaging in Example 1 was adjusted to examine the effects of 1.5 mL cryovials, 2 mL vials, and 10 mL vials on the labeling process. The other preparation processes of the freeze-dried powder preparation method remained unchanged.

[0137] (2) The corresponding reagent kit packaging contents are the same as those in Example 1.

[0138] (3)Al 18 The steps of F radiolabeling were the same as those in Example 1.

[0139] (4) The freeze-dried powder was prepared by adjusting the container used for final packaging and radiolabeled three batches continuously. The radiolabeling rates are shown in Table 6.

[0140] Table 6

[0141]

[0142]

[0143] Combined with the above table, among different reaction tube containers, 1.5mL plastic cryotubes have the lowest residual activity and the highest radioactive labeling rate. Compared with plastic cryotubes, glass vials have high residual activity, and as the volume of the glass vial increases, the residual activity increases and the labeling rate decreases. In summary, 1.5mL cryotubes are preferred as reaction tube containers.

[0144] Example 7

[0145] When the precursor is NOTA-PSMA-BCH, the research 18 The amount of F radioactive raw material affects the radioactive labeling rate. See Table 7 below for details:

[0146] (1) The freeze-dried powder preparation process is the same as that in Example 1.

[0147] (2) The corresponding reagent kit packaging contents are the same as those in Example 1.

[0148] (3)Al 18 The F radioactive labeling process is the same. By adjusting the accelerator bombardment time and beam intensity, it can produce radioactive activities of different 18 F radioactive material.

[0149] (4) Through adjustment 18 The starting dose of F radioactivity was 1.384 mmol / l and 1.5 mmol / l was used for continuous radiolabeling for 5 batches. The radiolabeling rates are shown in Table 7.

[0150] Table 7

[0151]

[0152]

[0153] Combined with the above table, within the activity range of 500mCi, the radioactive labeling rate is basically maintained at around 70%; when the activity is higher than 1000mCi, although Al 18 The high-dose preparation of F-PSMA-BCH was performed, but the radiolabeling rate was reduced to about 50%. In summary, under the premise of 100ug of precursor, the preferred 18 The radioactivity of F ions is within 500 mCi, which is a conventional small-dose, high-efficiency production; 18 The radioactivity of F ions is greater than 1000mCi, which is a large-dose production.

[0154] Example 8

[0155] Al 18 PET / CT imaging results of F-PSMA-BCH in prostate cancer patients:

[0156] Take the Al prepared in Example 1 18 About 10 mCi of F-PSMA-BCH was diluted with saline to 2 mL and injected intravenously into the patient. PET image data was acquired by static acquisition for 20 minutes 90 minutes after the injection. Figure 6 shown. Figure 6 It shows that Al 18F-PSMA-BCH has obvious high uptake in the primary lesion of the patient, and the double dashed arrow is the prostate tumor area; it also shows that Al 18 F-PSMA-BCH showed obvious bone metastatic lesions (single dashed arrows) with high contrast uptake in multiple parts of the patient's body, and there was also obvious uptake in the metastatic lymph nodes (single solid arrows).

[0157] Finally, it should be noted that: the applicant declares that the present invention uses the above-mentioned embodiments to illustrate the products and detailed preparation methods of the present invention, but the present invention is not limited to the above-mentioned products and detailed preparation methods, that is, it does not mean that the present invention must rely on the above-mentioned products and detailed preparation methods to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of the raw materials of the products of the present invention and the addition of auxiliary materials, the selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0158] The preferred embodiments of the present invention are described in detail above, but the present invention is not limited to the specific details in the above embodiments, and the technical solution of the present invention can be subjected to a variety of simple modifications within the technical concept of the present invention. These simple modifications all belong to the protection scope of the present invention.

[0159] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

[0160] In addition, various embodiments of the present invention may be arbitrarily combined, and as long as they do not violate the concept of the present invention, they should also be regarded as the contents disclosed by the present invention.

Claims

1. A kind of Al 18 The method for preparing the integrated freeze-dried powder of F tracer is characterized by the following steps: include: Dissolve the precursor with deionized water or 10-20% ethanol aqueous solution, then add pH buffer solution, AlCl3 solution, stabilizer solution, excipient, and finally add deionized water to make the concentration of the precursor 0.5-1.0 mM, mix the above solutions evenly and then divide them into packages, then place them in a freeze vacuum dryer to remove water at low temperature into powder or freeze-dried cake; The structural formula of the precursor contains a structural unit as shown in Formula 1: The molecular weight of the precursor is an Affibody, a polypeptide, a polypeptide-like structure or a small molecule structure not exceeding 10 KDa.

2. A kind of Al according to claim 1 18 The method for preparing the F tracer integrated freeze-dried powder is characterized in that: The precursor includes: any one of the polypeptides NOTA-PSMA-BCH, NOTA-DUPA-Pep, NOTA-FAPI-04, NOTA-FAPI-46, NOTA-FAPI-74, NOTA-FAPI-2286, NOTA-Octreotide, NOTA-TATE, NOTA-TOC, NOTA-JR11, NOTA-LM3, NOTA-AlfatideII, NOTA-Pentixafor, NOTA-DK222, NOTA-WL12, NOTA-PCP1, and NOTA-PCP2.

3. A kind of Al according to claim 1 18 The method for preparing the F tracer integrated freeze-dried powder is characterized in that: The pH of the pH buffer solution is 4.0-4.2, the concentration is 0.2-0.5M, and the volume dispensed into each container is 20-25uL.

4. A kind of Al according to claim 1 18 The method for preparing the F tracer integrated freeze-dried powder is characterized in that: The solute in the pH buffer solution is a non-volatile dicarboxylate or aminosulfonate, and the solute includes any one of succinic acid, glutaric acid, potassium hydrogen phthalate, 4-hydroxyethylpiperazineethanesulfonic acid and 2-morpholineethanesulfonic acid.

5. A kind of Al according to claim 1 18 The method for preparing the F tracer integrated freeze-dried powder is characterized in that: The concentration of the AlCl3 solution is 10 mM, the solvent is a 0.1 M acetic acid-sodium acetate buffer system with a pH value of 4.0, and the molar ratio of the amount of AlCl3 to the amount of the precursor in the mixed solution is 0.8-1.

0.

6. A kind of Al according to claim 1 18 The method for preparing the F tracer integrated freeze-dried powder is characterized in that: The stabilizer solution is an ascorbic acid aqueous solution or a gentisic acid aqueous solution with a concentration of 0.5 M, and the volume of the stabilizer solution dispensed into each container is 5-10 uL.

7. A kind of Al according to claim 1 18 The method for preparing the F tracer integrated freeze-dried powder is characterized in that: The excipient is any one of trehalose, mannitol and glucose, and the mass of the excipient packed into each container is 1-4 mg.

8. Aluminum 18 The kit of integrated lyophilized powder of F tracer is characterized in that: Comprising Al prepared by the preparation method according to any one of claims 1 to 7 18 F tracer integrated lyophilized powder, the integrated lyophilized powder is packaged in a 2mL vial or a 1.5mL cryotube, the vial or cryotube is not only a storage container for the lyophilized powder / cake, but also a reaction container for radioactive labeling.

9. The Al according to claim 8 18 The kit of integrated lyophilized powder of F tracer is characterized in that: The volume of the precursor solution dispensed into each tube or bottle is 100-200 uL.

10. Al prepared by the preparation method according to any one of claims 1 to 7 18 F tracer integrated freeze-dried powder and the Al according to claim 8 or 9 18 Application of the F tracer integrated freeze-dried powder kit in the preparation of PET / CT imaging agents.

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