A kind of Al 18 Preparation method, kit and application of F tracer integrated freeze-dried powder

Abstract

The present invention provides an Al 18 The preparation method, kit and application of an integrated F tracer lyophilized powder include: dissolving 1-10 mg of a precursor in deionized water, adding a pH buffer solution with a concentration of 0.2-0.5 M and an AlCl3 solution with a concentration of 10 mM, wherein the molar ratio of the AlCl3 to the precursor is 0.8-1.0, then adding 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 precursor concentration 0.5-1.0 mM. After uniformly mixing the above solutions, the solutions are dispensed into 1.5 mL cryovials or 2 mL vials according to a certain volume, and then placed in a freeze vacuum dryer to remove water at low temperature to obtain a powder or a lyophilized cake. The container serves as both a storage container for the lyophilized powder / cake and a reaction container for 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, easy central distribution, and long-distance transportation; high yield, usually ranging from a few hundred millicuries to a few curies; high image quality, high spatial resolution, and easy detection of tiny lesions. 18 The F marking method is basically based on 18 The F-labeled synthon is then coupled with a bioactive small molecule. Its disadvantages are that it is time-consuming and labor-intensive, with low yield, which is not conducive to clinical translation. In addition, due to structural changes, the activity of the bioactive molecule is changed, that is, the lipid-water partition coefficient is changed, which ultimately leads to a huge change 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, and finally form AlF-NOTA-ligand to achieve the effect 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 do not meet GMP requirements, and low yields 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 expensive price of disposable cartridge reagents 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 manually add them in sequence. This production process is cumbersome and prone to errors, resulting in tracer production failure. 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 The invention relates to a PET imaging agent, a preparation method and an application thereof, and belongs to the field of radiopharmaceuticals and nuclear medicine. The drug kit contains NOTA-PRGD2 and aluminum chloride, and the molar ratio of NOTA-PRGD2 to aluminum chloride is (1-10):1. The drug kit is in the form of a lyophilized powder. Preliminary studies have shown that the drug 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 appear 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 existence of 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 purpose 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 The F labeling rate is significantly improved compared to the traditional labeling process, making the labeling rate reach 65%-75%. The integrated freeze-dried powder / cake used in the present invention contains the 18 All components required for the F labeling process are contained in an EP tube or vial, which serves as both a container for the lyophilized powder and a reaction vessel. This significantly simplifies the tedious reagent filling process during radiolabeling preparation. Furthermore, compared to conventional procedures, it reduces material loss during the transfer of the precursor from the container to the reaction tube after dissolution.

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

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

[0010] The precursor is an affibody, a polypeptide, a polypeptide-like structure or a small molecule structure with a molecular weight 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 of the solution 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 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 0.5 M; and the volume of the stabilizer solution dispensed into each container is 5-10 uL.

[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] On the other hand, the present invention provides an Al 18 F tracer integrated lyophilized powder kit, the Al 18 The F tracer integrated freeze-dried powder is prepared by the above 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 sealed in 1.5 mL cryovials, then placed in a freeze vacuum dryer to remove water at low temperature into a lyophilized cake, which is then 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 cryovial serves as both a storage container for the lyophilized powder / cake and 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 not only can realize the freeze-drying of the freeze-dried drug 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 drug kit system does not change before and after the freeze-drying of the succinic acid system buffer, which facilitates 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, to prevent 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 chelate, thereby improving the labeling efficiency. In addition, stabilizers such as ascorbic acid or gentisic acid are added to the freeze-dried kit to prevent radiolytic decomposition, which can eliminate Al 18F labeling process of oxygen free radicals in the reaction system; at the same time, the addition of freeze-dried powder excipients does not affect the labeling rate to ensure the state and stability of the contents of the freeze-dried tube / bottle.

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

[0024] The kit provided by the present invention is used to perform AI 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, then add 18 F target water or 18 F concentrate is sufficient, and no additional reagents need to be added. Whether it is added automatically or manually, the tedious process of adding reagent solutions is eliminated, which greatly reduces the Al2O3 caused by human errors in the filling process. 18 F marks failure 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 currently known literature reports and patents, the technical effect (yield and output) of this invention is significant (currently usually Al 18 The F labeling rate is 30-40%, and the yield is usually in the range of tens to hundreds of millicuries. The implementation of this 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 smaller 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, resulting in waste and creating an additional burden on radiation protection.

[0026] By applying the kit and the corresponding preparation method of the present invention, the radioactive activity to which operators are directly exposed is reduced, and radiation damage to operators caused by unnecessary overexposure 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 Al of the present invention 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 a 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 following is a clear and complete description of the technical solution of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

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

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

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

[0038] Among them, the precursor includes any one of the 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 has the following structural formula:

[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 primarily used for the diagnosis of digestive system tumors. Its structural formula is as follows:

[0044]

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

[0046]

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

[0048]

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

[0050]

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

[0052]

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

[0054]

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

[0056]

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

[0058]

[0059] NOTA-LM3 targets the tumor somatostatin receptor and is mostly used in the diagnosis of neuroendocrine tumors. Its structural formula is as follows:

[0060]

[0061] NOTA-Alfatide II targets the αvβ3 receptor on tumor neovascularization and is mostly used in the diagnosis of non-small cell lung cancer. Its 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 PD-L1 receptor in the tumor immune microenvironment and has the following structural formula:

[0066]

[0067] NOTA-WL12 targets the PD-L1 receptor in the tumor immune microenvironment and has the following structural formula:

[0068]

[0069] NOTA-PCP1 targets the PD-L1 receptor in the tumor immune microenvironment and has the following structural formula:

[0070]

[0071] NOTA-PCP2 targets the tumor immune microenvironment PD-L1 receptor for imaging. The 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 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 of each cryotube or vial is 5-10uL; the excipient is any one of trehalose, mannitol, and glucose; and the mass of the excipient packed into each cryotube or vial is 1-4mg.

[0074] The present invention also provides an Al 18 F tracer integrated lyophilized powder kit, the Al 18 The F tracer integrated freeze-dried powder is prepared by the above method. The preparation method of the kit comprises the following steps: 18The integrated lyophilized powder of the F tracer is dispensed into 2 mL vials or packaged into 1.5 mL cryovials, then placed in a freeze vacuum dryer to remove water at low temperature into a lyophilized cake, which is then 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 cryovial serves as both a storage container for the lyophilized powder / cake and a reaction vessel for radioactive labeling. The volume of the precursor solution dispensed into each tube or bottle is 100-200 μL.

[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 its kit packaging contents, 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 μL of 10 mM AlCl3 solution, 250 μL 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 thoroughly mixed. The above solution was dispensed into 1.5 mL cryovials (30 tubes) and 3 mL vials (20 bottles) at a volume of 200 μL per tube, and placed in a freeze vacuum dryer to remove water at low temperature to form a freeze-dried cake. Figure 2 and Figure 3 );

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

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

[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 dried, and then rinsed with 120uL of normal saline and concentrated into a 1.5mL integrated freeze-drying tube.

[0084] (2) Then add 300uL DMF solvent into the reaction system, bubble and mix evenly.

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

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

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

[0088] (6) The solid phase extraction column was rinsed 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, 9 mL of normal saline was added for dilution. During the product elution process, the product was filtered through an online sterile filter membrane to prepare 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] Radiolabeling rate (%) = total activity of product solution (mCi) / 18 F initial activity (mCi)*100%;

[0091] Al prepared by the above process18 F-PSMA-BCH kit, continuous preparation of 3 batches of Al 18 The radiolabeling rate of F-PSMA-BCH tracer was between 65% and 75%, and the fluoride ion content was less than 3% as determined by radio-TLC. Figure 4 ), radiochemical purity was determined by Radio-HPLC>95% ( Figure 5 ), and the quality control parameters are detailed 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 radiolabeling rate was studied. Based on Example 1, the amount of the precursor used was 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. 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 μg are prepared, respectively.

[0098] (2) The contents of the corresponding kit package 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] Based on the above table, the labeling rate increases with the amount of precursor used. When the precursor dosage is greater than 100 μg, the labeling rate remains stable at around 75%. In summary, as a preferred method, the content of the lyophilized powder precursor is 100 μg, which can maintain a high radiolabeling rate.

[0105] Example 3

[0106] When the precursor is NOTA-PSMA-BCH, the effect of the type and amount of excipients on the radiolabeling rate was studied. Based on Example 1, the type and amount of excipients were 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 lyophilized powders with corresponding excipient contents of 1, 2, and 4 mg were prepared, respectively.

[0108] (2) The contents of the corresponding kit package 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 dosages, 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 had no significant effect on the labeling rate. In summary, the 4 mg dosage 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 radiolabeling rate was studied. Based on Example 1, the type and amount of organic solvent were 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 contents of the corresponding kit package are the same as those in Example 1.

[0119] (3)Al 18 During the radiolabeling process, the volume of organic solvent and water added to the cryopreservation tube was adjusted by changing the control program (see Table 4, Amount and Water Added columns for details). The remaining 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, and 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; under the premise of the same content of the above-mentioned organic solvents, there is no significant difference in the labeling rate; in summary, as a preference, DMSO or DMF is selected as the organic solvent added in the labeling process. Although acetonitrile and ethanol have similar labeling rates, 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 effects of buffer solution type and pH value on radiolabeling efficiency were studied. Based on Example 1, the buffer solution type was 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 those shown in Table 5. The other preparation processes remained unchanged, and lyophilized powders containing different buffer systems were prepared respectively.

[0128] (2) The contents of the corresponding kit package 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, and 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 combination of buffer salt and Al 3+ Significant chelation occurs, resulting in a low radiolabeling rate (<5%), making it unsuitable 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 drastic changes in pH, the labeling rates of 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 was 4.0 when the mixed solution was prepared, the buffer system contained volatile acetic acid components. During the freeze-drying process, the acetic acid volatilized, causing the pH value to change drastically after the freeze-dried powder was 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 process of the subsequent product injection solution. This substance is a non-endogenous substance related to plasticizers and poses potential biosafety issues).

[0134] Example 6

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

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

[0137] (2) The contents of the corresponding kit package 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) Lyophilized 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] Based on the above table, among different reaction tube containers, 1.5mL plastic cryotubes have the lowest residual activity and the highest radiolabeling rate. Compared with plastic cryotubes, glass vials have higher 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 the preferred reaction tube container.

[0144] Example 7

[0145] When the precursor is NOTA-PSMA-BCH, the research 18 The amount of F radioactive material affects the radiolabeling 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 contents of the corresponding kit package 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 different radioactive activities. 18 F radioactive raw materials.

[0149] (4) Through adjustment 18 The starting dose of F radioactivity was 1.58 wt %. Five batches were radiolabeled consecutively. The radiolabeling rates are shown in Table 7.

[0150] Table 7

[0151]

[0152]

[0153] Combined with the above table, when the activity range is within 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 and 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 normal saline to 2 mL and injected intravenously into the patient. PET image data was acquired 90 minutes after the injection for 20 minutes. Figure 6 shown. Figure 6 Indicates 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 has obvious bone metastases (single dashed arrows) in multiple locations throughout the patient's body, with high contrast uptake, and the metastatic lymph nodes also have obvious uptake (single solid arrows).

[0157] Finally, it should be noted that the applicant declares that while the above-described embodiments illustrate the products and detailed preparation methods of the present invention, the present invention is not limited to these products and detailed preparation methods. This does not imply that the present invention must rely on these products and detailed preparation methods for implementation. Those skilled in the art should understand that any improvements to the present invention, equivalent substitutions of raw materials for the products of the present invention, addition of auxiliary ingredients, and selection of specific methods, etc., fall within the scope of protection and disclosure 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 of the above embodiments. Within the technical concept of the present invention, various simple variations can be made to the technical solution of the present invention. These simple variations all fall within the scope of protection 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 appropriate manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

[0160] In addition, the 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 preparation method of F tracer integrated freeze-dried powder is characterized by the following steps: include: The precursor NOTA-PSMA-BCH is dissolved in deionized water or a 10-20% ethanol aqueous solution by volume, followed by the addition of a succinic acid pH buffer solution, an AlCl3 solution, an ascorbic acid stabilizer solution, and an excipient. Finally, deionized water is added to make the precursor concentration 0.5-1.0 mM. The above solutions are mixed evenly and then packaged. The solution is then placed in a freeze vacuum dryer to remove water at low temperature into a powder or freeze-dried cake. The pH of the pH buffer solution is 4.0-4.2, the concentration is 0.5 M, and the volume of the solution dispensed into each container is 20-25 uL.

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

3. A kind of Al according to claim 1 18 The preparation method of 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.

4. A kind of Al 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 3 18 F tracer integrated lyophilized powder, the integrated lyophilized powder is packaged in a 2 mL vial or a 1.5 mL cryotube, the vial or cryotube serves as both a storage container for the lyophilized powder / cake and a reaction container for radioactive labeling.

5. Al according to claim 4 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.

6. Al prepared by the preparation method according to any one of claims 1 to 3 18 F tracer integrated freeze-dried powder and the Al according to claim 4 or 5 18 Application of the integrated freeze-dried powder kit of F tracer in the preparation of PET / CT imaging agents.

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